Datasets:

tomekkorbak

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detoxify-pile-chunk3-1350000-1400000

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[ "Blue Marsh National Recreation Area\n\nBlue Marsh National Recreation Area is an artificial lake located northwest of the city of Reading, Pennsylvania, USA and managed by the U.S. Army Corps of Engineers Philadelphia District. ", "It is in western Berks County, fed into by the Tulpehocken Creek. ", "The main span of the lake is along the border between Bern and Lower Heidelberg Townships. ", "However, the northwesternmost portions lie in the more sparsely populated North Heidelberg and Penn Townships. ", "In the middle of the lake is a large, uninhabited island (claimed by Bern Township). ", "The lake is a popular recreation area in the summer, where people can fish, swim, and boat. ", "It has 36 miles of trails and 1,147 acres of water.", "\n\nHistory\nBlue Marsh was the name of the village that was located where the lake now is. ", "It was the first settlement in Lower Heidelberg Township. ", "The land was very fertile. ", "It was also a heavily forested area with abundance of wildlife. ", "There were many farms and 18th and 19th century homes. ", "The village also had a church, schoolhouse, post office, and Pyles General Store. ", "Some buildings were spared, such as Gruber Wagon Works, which was originally in Blue Marsh but was moved a safe distance away before the dam was created. ", "Others include Old Dry Rd. ", "Farm, which most of its buildings were relocated out of the village. ", "Residents were put under eminent domain and had no choice but to move out. ", "Initial authorization for the reservoir was granted by the Flood Control Act of 1962 that started eight projects on the Delaware River basin. ", " The Pennsylvania Project 70 Land Acquisition and Borrowing Act provided funding and permitted the eminent domain acquisition of the land that would later become the lake with the governor's permission in 1969. ", " The United States Army Corps of Engineers began constructing the lake in March 1974 with the impoundment of the Tulpehocken Creek and was completed in September 1979.", "\n\nReferences\n\nExternal links\n , official website from the U.S. Army Corps of Engineers\n\nCategory:Reservoirs in Pennsylvania\nCategory:Protected areas of Berks County, Pennsylvania\nCategory:Bodies of water of Berks County, Pennsylvania\nCategory:Protected areas established in 1979\nCategory:1979 establishments in Pennsylvania" ]

{ "pile_set_name": "Wikipedia (en)" }

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[ "/**\r\n * \\file\r\n *\r\n * Copyright (c) 2015 Atmel Corporation. ", "All rights reserved.", "\r\n *\r\n * \\asf_license_start\r\n *\r\n * \\page License\r\n *\r\n * Redistribution and use in source and binary forms, with or without\r\n * modification, are permitted provided that the following conditions are met:\r\n *\r\n * 1. ", "Redistributions of source code must retain the above copyright notice,\r\n * this list of conditions and the following disclaimer.", "\r\n *\r\n * 2. ", "Redistributions in binary form must reproduce the above copyright notice,\r\n * this list of conditions and the following disclaimer in the documentation\r\n * and/or other materials provided with the distribution.", "\r\n *\r\n * 3. ", "The name of Atmel may not be used to endorse or promote products derived\r\n * from this software without specific prior written permission.", "\r\n *\r\n * 4. ", "This software may only be redistributed and used in connection with an\r\n * Atmel microcontroller product.", "\r\n *\r\n * THIS SOFTWARE IS PROVIDED BY ATMEL \"AS IS\" AND ANY EXPRESS OR IMPLIED\r\n * WARRANTIES, INCLUDING, BUT NOT LIMITED TO, THE IMPLIED WARRANTIES OF\r\n * MERCHANTABILITY, FITNESS FOR A PARTICULAR PURPOSE AND NON-INFRINGEMENT ARE\r\n * EXPRESSLY AND SPECIFICALLY DISCLAIMED. ", "IN NO EVENT SHALL ATMEL BE LIABLE FOR\r\n * ANY DIRECT, INDIRECT, INCIDENTAL, SPECIAL, EXEMPLARY, OR CONSEQUENTIAL\r\n * DAMAGES (INCLUDING, BUT NOT LIMITED TO, PROCUREMENT OF SUBSTITUTE GOODS\r\n * OR SERVICES; LOSS OF USE, DATA, OR PROFITS; OR BUSINESS INTERRUPTION)\r\n * HOWEVER CAUSED AND ON ANY THEORY OF LIABILITY, WHETHER IN CONTRACT,\r\n * STRICT LIABILITY, OR TORT (INCLUDING NEGLIGENCE OR OTHERWISE) ARISING IN\r\n * ANY WAY OUT OF THE USE OF THIS SOFTWARE, EVEN IF ADVISED OF THE\r\n * POSSIBILITY OF SUCH DAMAGE.", "\r\n *\r\n * \\asf_license_stop\r\n *\r\n */\r\n/*\r\n * Support and FAQ: visit <a href=\"http://www.atmel.com/design-support/\">Atmel Support</a>\r\n */\r\n\r\n#ifndef _SAMV71_SSC_INSTANCE_\r\n#define _SAMV71_SSC_INSTANCE_\r\n\r\n/* ========== Register definition for SSC peripheral ========== */\r\n#if (defined(__ASSEMBLY__) || defined(__IAR_SYSTEMS_ASM__))\r\n #define REG_SSC_CR (0x40004000U) /**< \\brief (SSC) Control Register */\r\n #define REG_SSC_CMR (0x40004004U) /**< \\brief (SSC) Clock Mode Register */\r\n #define REG_SSC_RCMR (0x40004010U) /**< \\brief (SSC) Receive Clock Mode Register */\r\n #define REG_SSC_RFMR (0x40004014U) /**< \\brief (SSC) Receive Frame Mode Register */\r\n #define REG_SSC_TCMR (0x40004018U) /**< \\brief (SSC) Transmit Clock Mode Register */\r\n #define REG_SSC_TFMR (0x4000401CU) /**< \\brief (SSC) Transmit Frame Mode Register */\r\n #define REG_SSC_RHR (0x40004020U) /**< \\brief (SSC) Receive Holding Register */\r\n #define REG_SSC_THR (0x40004024U) /**< \\brief (SSC) Transmit Holding Register */\r\n #define REG_SSC_RSHR (0x40004030U) /**< \\brief (SSC) Receive Sync. ", "Holding Register */\r\n #define REG_SSC_TSHR (0x40004034U) /**< \\brief (SSC) Transmit Sync. ", "Holding Register */\r\n #define REG_SSC_RC0R (0x40004038U) /**< \\brief (SSC) Receive Compare 0 Register */\r\n #define REG_SSC_RC1R (0x4000403CU) /**< \\brief (SSC) Receive Compare 1 Register */\r\n #define REG_SSC_SR (0x40004040U) /**< \\brief (SSC) Status Register */\r\n #define REG_SSC_IER (0x40004044U) /**< \\brief (SSC) Interrupt Enable Register */\r\n #define REG_SSC_IDR (0x40004048U) /**< \\brief (SSC) Interrupt Disable Register */\r\n #define REG_SSC_IMR (0x4000404CU) /**< \\brief (SSC) Interrupt Mask Register */\r\n #define REG_SSC_WPMR (0x400040E4U) /**< \\brief (SSC) Write Protection Mode Register */\r\n #define REG_SSC_WPSR (0x400040E8U) /**< \\brief (SSC) Write Protection Status Register */\r\n#else\r\n #define REG_SSC_CR (*(__O uint32_t*)0x40004000U) /**< \\brief (SSC) Control Register */\r\n #define REG_SSC_CMR (*(__IO uint32_t*)0x40004004U) /**< \\brief (SSC) Clock Mode Register */\r\n #define REG_SSC_RCMR (*(__IO uint32_t*)0x40004010U) /**< \\brief (SSC) Receive Clock Mode Register */\r\n #define REG_SSC_RFMR (*(__IO uint32_t*)0x40004014U) /**< \\brief (SSC) Receive Frame Mode Register */\r\n #define REG_SSC_TCMR (*(__IO uint32_t*)0x40004018U) /**< \\brief (SSC) Transmit Clock Mode Register */\r\n #define REG_SSC_TFMR (*(__IO uint32_t*)0x4000401CU) /**< \\brief (SSC) Transmit Frame Mode Register */\r\n #define REG_SSC_RHR (*(__I uint32_t*)0x40004020U) /**< \\brief (SSC) Receive Holding Register */\r\n #define REG_SSC_THR (*(__O uint32_t*)0x40004024U) /**< \\brief (SSC) Transmit Holding Register */\r\n #define REG_SSC_RSHR (*(__I uint32_t*)0x40004030U) /**< \\brief (SSC) Receive Sync. ", "Holding Register */\r\n #define REG_SSC_TSHR (*(__IO uint32_t*)0x40004034U) /**< \\brief (SSC) Transmit Sync. ", "Holding Register */\r\n #define REG_SSC_RC0R (*(__IO uint32_t*)0x40004038U) /**< \\brief (SSC) Receive Compare 0 Register */\r\n #define REG_SSC_RC1R (*(__IO uint32_t*)0x4000403CU) /**< \\brief (SSC) Receive Compare 1 Register */\r\n #define REG_SSC_SR (*(__I uint32_t*)0x40004040U) /**< \\brief (SSC) Status Register */\r\n #define REG_SSC_IER (*(__O uint32_t*)0x40004044U) /**< \\brief (SSC) Interrupt Enable Register */\r\n #define REG_SSC_IDR (*(__O uint32_t*)0x40004048U) /**< \\brief (SSC) Interrupt Disable Register */\r\n #define REG_SSC_IMR (*(__I uint32_t*)0x4000404CU) /**< \\brief (SSC) Interrupt Mask Register */\r\n #define REG_SSC_WPMR (*(__IO uint32_t*)0x400040E4U) /**< \\brief (SSC) Write Protection Mode Register */\r\n #define REG_SSC_WPSR (*(__I uint32_t*)0x400040E8U) /**< \\brief (SSC) Write Protection Status Register */\r\n#endif /* (defined(__ASSEMBLY__) || defined(__IAR_SYSTEMS_ASM__)) */\r\n\r\n#endif /* _SAMV71_SSC_INSTANCE_ */\r\n" ]

{ "pile_set_name": "Github" }

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[ "Q:\n\nWhat kind/kinds of car/cars are you interested in?", "\n\nWhat kind of car are you interested in? ", " \nWhat kind of cars are you interested in? ", " \nWhat kinds of cars are you interested in? ", " \n\nIs there any one that can’t be allowed to be said? ", " \nI would think that #1 could be possible, when the speakers, intending to buy a car, are in a shop which sells cars. ", "And in #2, when it’s not the case as 1, plural cars are more reasonable, in my eye. ", "In 3, for ‘cars’ is plural, it seems that plural ‘kinds’ can be used - but sounds somewhat strange. ", "Can all three be said or not?", "\n\nA:\n\nYour understanding of #1 is correct: someone shopping for a car is asked \"what kind of car are you shopping for, family sedan, small commuting car, a big station-wagon or SUV?\"", "\nStatement #3 might apply to a situation where a collector is asked to list the range of vehicle types he likes to collect (racing cars, luxury cars, cars formerly owned by celebrities, etc).", "\nStatement #2 might apply to a situation where a highly specialized collector, who collects only one kind of vehicle, is asked to say which kind that is (sports cars or luxury cars or cars whose body panels were made of soybeans...). ", " The question assumes a single property will unite these cars into a class. ", "The collector might say he collects \"only vintage cars\" but they could include vintage racing cars and vintage luxury cars and ...\n\n" ]

{ "pile_set_name": "StackExchange" }

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[ "Q:\n\nWhen should a type be a struct containing another type and when should it just \"extend\"(?) ", "that type?", "\n\nI'm currently learning Go by doing the rosalind problems (basically a bunch of bioinformatics related code katas). ", "\nI'm currently representing a DNA strand with a type:\ntype DNAStrand struct {\n dna byte[]\n}\n\nMy initial reason was to encapsulate the byte slice so I would know it only contained bytes representing the nucleotides: 'A', 'C', 'G', 'T'. ", "I realized that this was obviously not guarateed since I could simply do:\nDNAStrand{[]byte(\"foo bar\")}\n\nAnd there is no longer any guarantee that my dna strand contains a byte array with only elements from those four bytes.", "\nSince my struct only contains a byte array is it better/more ideomatic to do:\ntype DNAStrand []byte\n\nOr is it better to let the type contain the dna strand? ", "Are there any rules of thumb for when to use either of the two approaches?", "\n\nA:\n\nTaking your specific example I would probably do something like this:\ntype neucleotide char // unexported type users can't construct their own.", "\n\ntype DNAStrand []neucleotide // because users can't construct their own\n // nucleotides they also can't construct their\n // own DNAStrands.", "\n\nconst (\n // These are exported values so they can use these nucleotides to construct a\n // DNAStrand with.", "\n A nucleotide = 'A'\n C nucleotide = 'C'\n G nudleotide = 'G'\n T nucleotide = 'T'\n)\n\n// This function allows them to actually construct a DNAstrand with a list of\n// nucleotides from the constants above.", "\nfunc New(nts ...nucleotide) DNAStrand {\n return nts\n}\n\nSince the nucleotide type is not exported users can't construct their own. ", "You provide the only allowed instances of them in the exported consts so no user can provide their own new nucleotides.", "\n\nA:\n\nStruct with zero fields are handy. ", "Structs with many fields are handy even more. ", "Structs with exactly one field are a bit special and I can't think of a reasonably \"good\" case where to use them - even though they are seen regularly \"in the wild\". ", "I, for one, don't use them.", "\nAnyway, if you really really need tighter/bulletproof safety about the DNAStrand slice content - then it is possible to to use the single field struct and define an argument checking setter method for this/such named type.", "\nIn that case, if the definition is later used from some other package, there's no way, modulo using package unsafe, to circumvent the checks and get a result equivalent to your DNAStrand{[]byte(\"foo bar\")} example.", "\n\n" ]

{ "pile_set_name": "StackExchange" }

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[ "//\n// _RXObjCRuntime.m\n// RxCocoa\n//\n// Created by Krunoslav Zaher on 7/11/15.", "\n// Copyright © 2015 Krunoslav Zaher. ", "All rights reserved.", "\n//\n\n#import <pthread.h>\n#import <Foundation/Foundation.h>\n#import <objc/runtime.h>\n#import <objc/message.h>\n#import <libkern/OSAtomic.h>\n\n#import \"_RX.h\"\n#import \"_RXObjcRuntime.h\"\n\n#if !", "DISABLE_SWIZZLING\n\n// self + cmd\n#define HIDDEN_ARGUMENT_COUNT 2\n\n@class RXObjCRuntime;\n\nBOOL RXAbortOnThreadingHazard = NO;\n\ntypedef NSInvocation *NSInvocationRef;\ntypedef NSMethodSignature *NSMethodSignatureRef;\ntypedef unsigned char rx_uchar;\ntypedef unsigned short rx_ushort;\ntypedef unsigned int rx_uint;\ntypedef unsigned long rx_ulong;\ntypedef id (^rx_block)(id);\ntypedef BOOL (^RXInterceptWithOptimizedObserver)(RXObjCRuntime * __nonnull self, Class __nonnull class, SEL __nonnull selector, NSError ** __nonnull error);\n\nstatic CFTypeID defaultTypeID;\nstatic SEL deallocSelector;\n\nstatic int RxSwizzlingTargetClassKey = 0;\n\n#if TRACE_RESOURCES\nstatic int32_t numberOInterceptedMethods = 0;\nstatic int32_t numberOfForwardedMethods = 0;\n#endif\n\n#define THREADING_HAZARD(class) \\\n NSLog(@\"There was a problem swizzling on `%@`.\\nYou have probably two libraries performing swizzling in runtime.\\nWe didn't want to crash your program, but this is not good ...\\nYou an solve this problem by either not using swizzling in this library, removing one of those other libraries, or making sure that swizzling parts are synchronized (only perform them on main thread).\\nAnd yes, this message will self destruct when you clear the console, and since it's non deterministic, the problem could still exist and it will be hard for you to reproduce it.\", ", "NSStringFromClass(class)); ABORT_IN_DEBUG if (RXAbortOnThreadingHazard) { abort(); }\n\n#define ALWAYS(condition, message) if (!(", "condition)) { [NSException raise:@\"RX Invalid Operator\" format:@\"%@\", message]; }\n#define ALWAYS_WITH_INFO(condition, message) NSAssert((condition), @\"%@ [%@] > %@\", NSStringFromClass(class), NSStringFromSelector(selector), (message))\n#define C_ALWAYS(condition, message) NSCAssert((condition), @\"%@ [%@] > %@\", NSStringFromClass(class), NSStringFromSelector(selector), (message))\n\n#define RX_PREFIX @\"_RX_namespace_\"\n\n#define RX_ARG_id(value) ((value) ?: [", "NSNull null])\n#define RX_ARG_char(value) [NSNumber numberWithChar:value]\n#define RX_ARG_short(value) [NSNumber numberWithShort:value]\n#define RX_ARG_int(value) [NSNumber numberWithInt:value]\n#define RX_ARG_long(value) [NSNumber numberWithLong:value]\n#define RX_ARG_BOOL(value) [NSNumber numberWithBool:value]\n#define RX_ARG_SEL(value) [NSNumber valueWithPointer:value]\n#define RX_ARG_rx_uchar(value) [NSNumber numberWithUnsignedInt:value]\n#define RX_ARG_rx_ushort(value) [NSNumber numberWithUnsignedInt:value]\n#define RX_ARG_rx_uint(value) [NSNumber numberWithUnsignedInt:value]\n#define RX_ARG_rx_ulong(value) [NSNumber numberWithUnsignedLong:value]\n#define RX_ARG_rx_block(value) ((id)(value) ?: [", "NSNull null])\n#define RX_ARG_float(value) [NSNumber numberWithFloat:value]\n#define RX_ARG_double(value) [NSNumber numberWithDouble:value]\n\ntypedef struct supported_type {\n const char *encoding;\n} supported_type_t;\n\nstatic supported_type_t supported_types[] = {\n { .encoding = @encode(void)},\n { .encoding = @encode(id)},\n { .encoding = @encode(Class)},\n { .encoding = @encode(void (^)())},\n { .encoding = @encode(char)},\n { .encoding = @encode(short)},\n { .encoding = @encode(int)},\n { .encoding = @encode(long)},\n { .encoding = @encode(long long)},\n { .encoding = @encode(unsigned char)},\n { .encoding = @encode(unsigned short)},\n { .encoding = @encode(unsigned int)},\n { .encoding = @encode(unsigned long)},\n { .encoding = @encode(unsigned long long)},\n { .encoding = @encode(float)},\n { .encoding = @encode(double)},\n { .encoding = @encode(BOOL)},\n { .encoding = @encode(const char*)},\n};\n\nNSString * __nonnull const RXObjCRuntimeErrorDomain = @\"RXObjCRuntimeErrorDomain\";\nNSString * __nonnull const RXObjCRuntimeErrorIsKVOKey = @\"RXObjCRuntimeErrorIsKVOKey\";\n\nBOOL RX_return_type_is_supported(const char *type) {\n if (type == nil) {\n return NO;\n }\n\n for (int i = 0; i < sizeof(supported_types) / sizeof(supported_type_t); ++i) {\n if (supported_types[i].encoding[0] !", "= type[0]) {\n continue;\n }\n if (strcmp(supported_types[i].encoding, type) == 0) {\n return YES;\n }\n }\n\n return NO;\n}\n\nstatic BOOL RX_method_has_supported_return_type(Method method) {\n const char *rawEncoding = method_getTypeEncoding(method);\n ALWAYS(rawEncoding !", "= nil, @\"Example encoding method is nil.\");", "\n\n NSMethodSignature *methodSignature = [NSMethodSignature signatureWithObjCTypes:rawEncoding];\n ALWAYS(methodSignature !", "= nil, @\"Method signature method is nil.\");", "\n\n return RX_return_type_is_supported(methodSignature.methodReturnType);\n}\n\nSEL __nonnull RX_selector(SEL __nonnull selector) {\n NSString *selectorString = NSStringFromSelector(selector);\n return NSSelectorFromString([RX_PREFIX stringByAppendingString:selectorString]);\n}\n\nBOOL RX_is_method_signature_void(NSMethodSignature * __nonnull methodSignature) {\n const char *methodReturnType = methodSignature.methodReturnType;\n return strcmp(methodReturnType, @encode(void)) == 0;\n}\n\nBOOL RX_is_method_with_description_void(struct objc_method_description method) {\n return strncmp(method.types, @encode(void), 1) == 0;\n}\n\n// inspired by https://github.com/ReactiveCocoa/ReactiveCocoa/blob/swift-development/ReactiveCocoa/Objective-C/NSInvocation%2BRACTypeParsing.m\n// awesome work\nid __nonnull RX_extract_argument_at_index(NSInvocation * __nonnull invocation, NSUInteger index) {\n const char *argumentType = [invocation.methodSignature getArgumentTypeAtIndex:index];\n \n#define RETURN_VALUE(type) \\\n else if (strcmp(argumentType, @encode(type)) == 0) {\\\n type val = 0; \\\n [invocation getArgument:&val atIndex:index]; \\\n return @(val); \\\n }\n\n // Skip const type qualifier.", "\n if (argumentType[0] == 'r') {\n argumentType++;\n }\n \n if (strcmp(argumentType, @encode(id)) == 0\n || strcmp(argumentType, @encode(Class)) == 0\n || strcmp(argumentType, @encode(void (^)())) == 0\n ) {\n __unsafe_unretained id argument = nil;\n [invocation getArgument:&argument atIndex:index];\n return argument;\n }\n RETURN_VALUE(char)\n RETURN_VALUE(short)\n RETURN_VALUE(int)\n RETURN_VALUE(long)\n RETURN_VALUE(long long)\n RETURN_VALUE(unsigned char)\n RETURN_VALUE(unsigned short)\n RETURN_VALUE(unsigned int)\n RETURN_VALUE(unsigned long)\n RETURN_VALUE(unsigned long long)\n RETURN_VALUE(float)\n RETURN_VALUE(double)\n RETURN_VALUE(BOOL)\n RETURN_VALUE(const char *)\n else {\n NSUInteger size = 0;\n NSGetSizeAndAlignment(argumentType, &size, NULL);\n NSCParameterAssert(size > 0);\n uint8_t data[size];\n [invocation getArgument:&data atIndex:index];\n \n return [NSValue valueWithBytes:&data objCType:argumentType];\n }\n}\n\nNSArray *RX_extract_arguments(NSInvocation *invocation) {\n NSUInteger numberOfArguments = invocation.methodSignature.numberOfArguments;\n NSUInteger numberOfVisibleArguments = numberOfArguments - HIDDEN_ARGUMENT_COUNT;\n \n NSCParameterAssert(numberOfVisibleArguments >= 0);\n \n NSMutableArray *arguments = [NSMutableArray arrayWithCapacity:numberOfVisibleArguments];\n \n for (NSUInteger index = HIDDEN_ARGUMENT_COUNT; index < numberOfArguments; ++index) {\n [arguments addObject:RX_extract_argument_at_index(invocation, index) ?: [", "NSNull null]];\n }\n \n return arguments;\n}\n\nvoid * __nonnull RX_reference_from_selector(SEL __nonnull selector) {\n return selector;\n}\n\nstatic BOOL RX_forward_invocation(id __nonnull __unsafe_unretained self, NSInvocation *invocation) {\n SEL originalSelector = RX_selector(invocation.selector);\n\n id<RXMessageSentObserver> messageSentObserver = objc_getAssociatedObject(self, originalSelector);\n\n if (messageSentObserver !", "= nil) {\n NSArray *arguments = RX_extract_arguments(invocation);\n [messageSentObserver messageSentWithParameters:arguments];\n }\n\n if ([self respondsToSelector:originalSelector]) {\n invocation.selector = originalSelector;\n [invocation invokeWithTarget:self];\n return YES;\n }\n\n return NO;\n}\n\nstatic BOOL RX_responds_to_selector(id __nonnull __unsafe_unretained self, SEL selector) {\n Class class = object_getClass(self);\n if (class == nil) { return NO; }\n\n Method m = class_getInstanceMethod(class, selector);\n return m !", "= nil;\n\n}\n\nstatic NSMethodSignatureRef RX_method_signature(id __nonnull __unsafe_unretained self, SEL selector) {\n Class class = object_getClass(self);\n if (class == nil) { return nil; }\n\n Method method = class_getInstanceMethod(class, selector);\n if (method == nil) { return nil; }\n\n const char *encoding = method_getTypeEncoding(method);\n if (encoding == nil) { return nil; }\n\n return [NSMethodSignature signatureWithObjCTypes:encoding];\n}\n\nstatic NSString * __nonnull RX_method_encoding(Method __nonnull method) {\n const char *typeEncoding = method_getTypeEncoding(method);\n ALWAYS(typeEncoding !", "= nil, @\"Method encoding is nil.\");", "\n\n NSString *encoding = [NSString stringWithCString:typeEncoding encoding:NSASCIIStringEncoding];\n ALWAYS(encoding !", "= nil, @\"Can't convert encoding to NSString.\");", "\n return encoding;\n}\n\n// inspired by\n// https://github.com/mikeash/MAZeroingWeakRef/blob/master/Source/MAZeroingWeakRef.m\n// https://github.com/ReactiveCocoa/ReactiveCocoa/blob/swift-development/ReactiveCocoa/Objective-C/NSObject%2BRACDeallocating.m\n// https://github.com/steipete/Aspects\n@interface RXObjCRuntime: NSObject\n\n@property (nonatomic, assign) pthread_mutex_t lock;\n\n@property (nonatomic, strong) NSMutableSet<NSValue *> *classesThatSupportObservingByForwarding;\n@property (nonatomic, strong) NSMutableDictionary<NSValue *, NSMutableSet<NSValue*> *> *forwardedSelectorsByClass;\n\n@property (nonatomic, strong) NSMutableDictionary<NSValue *, Class> *dynamicSubclassByRealClass;\n@property (nonatomic, strong) NSMutableDictionary<NSValue *, NSMutableDictionary<NSValue*, NSValue *>*> *interceptorIMPbySelectorsByClass;\n\n+(RXObjCRuntime*)instance;\n\n-(void)performLocked:(void (^)(RXObjCRuntime* __nonnull))action;\n-(IMP __nullable)ensurePrepared:(id __nonnull)target forObserving:(SEL __nonnull)selector error:(NSError** __nonnull)error;\n-(BOOL)ensureSwizzledSelector:(SEL __nonnull)selector\n ofClass:(Class __nonnull)class\n newImplementationGenerator:(IMP(^)())newImplementationGenerator\nreplacementImplementationGenerator:(IMP (^)(IMP originalImplementation))replacementImplementationGenerator\n error:(NSError ** __nonnull)error;\n\n\n+(void)registerOptimizedObserver:(RXInterceptWithOptimizedObserver)registration encodedAs:(SEL)selector;\n\n@end\n\n/**\n All API methods perform work on locked instance of `RXObjCRuntime`. ", "In that way it's easy to prove\n that every action is properly locked.", "\n */\nIMP __nullable RX_ensure_observing(id __nonnull target, SEL __nonnull selector, NSError ** __nonnull error) {\n __block IMP __nonnull targetImplementation = nil;\n // Target is the second object that needs to be synchronized to TRY to make sure other swizzling framework\n // won't do something in parallel.", "\n // Even though this is too fine grained locking and more coarse grained locks should exist, this is just in case\n // someone calls this method directly without any external lock.", "\n @synchronized(target) {\n // The only other resource that all other swizzling libraries have in common without introducing external\n // dependencies is class object.", "\n //\n // It is polite to try to synchronize it in hope other unknown entities will also attempt to do so.", "\n // It's like trying to figure out how to communicate with aliens without actually communicating,\n // save for the fact that aliens are people, programmers, authors of swizzling libraries.", "\n @synchronized([target class]) {\n [[RXObjCRuntime instance] performLocked:^(RXObjCRuntime * __nonnull self) {\n targetImplementation = [self ensurePrepared:target\n forObserving:selector\n error:error];\n }];\n }\n }\n\n return targetImplementation;\n}\n\nIMP __nonnull RX_default_target_implementation() {\n return _objc_msgForward;\n}\n\n// bodies\n\n#define FORWARD_BODY(invocation) if (RX_forward_invocation(self, NAME_CAT(_, 0, invocation))) { return; }\n\n#define RESPONDS_TO_SELECTOR_BODY(selector) if (RX_responds_to_selector(self, NAME_CAT(_, 0, selector))) return YES;\n\n#define CLASS_BODY(...) return actAsClass;\n\n#define METHOD_SIGNATURE_FOR_SELECTOR_BODY(selector) \\\n NSMethodSignatureRef methodSignature = RX_method_signature(self, NAME_CAT(_, 0, selector)); \\\n if (methodSignature !", "= nil) { \\\n return methodSignature; \\\n }\n\n#define DEALLOCATING_BODY(...) \\\n id<RXMessageSentObserver> observer = objc_getAssociatedObject(self, rxSelector); \\\n if (observer !", "= nil && observer.targetImplementation == thisIMP) { \\\n [observer messageSentWithParameters:@[]]; \\\n }\n\n#define OBSERVE_BODY(...) \\\n id<RXMessageSentObserver> observer = objc_getAssociatedObject(self, rxSelector); \\\n \\\n if (observer !", "= nil && observer.targetImplementation == thisIMP) { \\\n [observer messageSentWithParameters:@[COMMA_DELIMITED_ARGUMENTS(__VA_ARGS__)]]; \\\n } \\\n\n\n#define BUILD_ARG_WRAPPER(type) RX_ARG_ ## type //RX_ARG_ ## type\n\n#define CAT(_1, _2, head, tail) RX_CAT2(head, tail)\n#define SEPARATE_BY_COMMA(_1, _2, head, tail) head, tail\n#define SEPARATE_BY_SPACE(_1, _2, head, tail) head tail\n#define SEPARATE_BY_UNDERSCORE(head, tail) RX_CAT2(RX_CAT2(head, _), tail)\n\n#define UNDERSCORE_TYPE_CAT(_1, index, type) RX_CAT2(_, type) // generates -> _type\n#define NAME_CAT(_1, index, type) SEPARATE_BY_UNDERSCORE(type, index) // generates -> type_0\n#define TYPE_AND_NAME_CAT(_1, index, type) type SEPARATE_BY_UNDERSCORE(type, index) // generates -> type type_0\n#define NOT_NULL_ARGUMENT_CAT(_1, index, type) BUILD_ARG_WRAPPER(type)(NAME_CAT(_1, index, type)) // generates -> ((id)(type_0) ?: [", "NSNull null])\n#define EXAMPLE_PARAMETER(_1, index, type) RX_CAT2(_, type):(type)SEPARATE_BY_UNDERSCORE(type, index) // generates -> _type:(type)type_0\n#define SELECTOR_PART(_1, index, type) RX_CAT2(_, type:) // generates -> _type:\n\n#define COMMA_DELIMITED_ARGUMENTS(...) RX_FOR(_, SEPARATE_BY_COMMA, NOT_NULL_ARGUMENT_CAT, ## __VA_ARGS__)\n#define ARGUMENTS(...) RX_FOR_COMMA(_, NAME_CAT, ## __VA_ARGS__)\n#define DECLARE_ARGUMENTS(...) RX_FOR_COMMA(_, TYPE_AND_NAME_CAT, ## __VA_ARGS__)\n\n// optimized observe methods\n\n#define GENERATE_METHOD_IDENTIFIER(...) RX_CAT2(swizzle, RX_FOR(_, CAT, UNDERSCORE_TYPE_CAT, ## __VA_ARGS__))\n\n#define GENERATE_OBSERVE_METHOD_DECLARATION(...) \\\n -(BOOL)GENERATE_METHOD_IDENTIFIER(__VA_ARGS__):(Class __nonnull)class \\\n selector:(SEL)selector \\\n error:(NSError ** __nonnull)error { \\\n\n\n#define BUILD_EXAMPLE_METHOD(return_value, ...) \\\n +(return_value)RX_CAT2(RX_CAT2(example_, return_value), RX_FOR(_, SEPARATE_BY_SPACE, EXAMPLE_PARAMETER, ## __VA_ARGS__)) {}\n\n#define BUILD_EXAMPLE_METHOD_SELECTOR(return_value, ...) \\\n RX_CAT2(RX_CAT2(example_, return_value), RX_FOR(_, SEPARATE_BY_SPACE, SELECTOR_PART, ## __VA_ARGS__))\n\n#define SWIZZLE_OBSERVE_METHOD(return_value, ...) \\\n @interface RXObjCRuntime (GENERATE_METHOD_IDENTIFIER(return_value, ## __VA_ARGS__)) \\\n @end \\\n \\\n @implementation RXObjCRuntime(GENERATE_METHOD_IDENTIFIER(return_value, ## __VA_ARGS__)) \\\n BUILD_EXAMPLE_METHOD(return_value, ## __VA_ARGS__) \\\n SWIZZLE_METHOD(return_value, GENERATE_OBSERVE_METHOD_DECLARATION(return_value, ## __VA_ARGS__), OBSERVE_BODY, ## __VA_ARGS__) \\\n \\\n +(void)load { \\\n __unused SEL exampleSelector = @selector(BUILD_EXAMPLE_METHOD_SELECTOR(return_value, ## __VA_ARGS__)); \\\n [self registerOptimizedObserver:^BOOL(RXObjCRuntime * __nonnull self, Class __nonnull class, \\\n SEL __nonnull selector, NSError **__nonnull error) { \\\n return [self GENERATE_METHOD_IDENTIFIER(return_value, ## __VA_ARGS__):class selector:selector error:error]; \\\n } encodedAs:exampleSelector]; \\\n } \\\n \\\n @end \\\n\n// infrastructure method\n\n#define SWIZZLE_INFRASTRUCTURE_METHOD(return_value, method_name, parameters, method_selector, body, ...) \\\n SWIZZLE_METHOD(return_value, -(BOOL)method_name:(Class __nonnull)class parameters error:(NSError **__nonnull)error \\\n { \\\n SEL selector = method_selector; , body, __VA_ARGS__) \\\n\n\n// common base\n\n#define SWIZZLE_METHOD(return_value, method_prototype, body, ...) \\\nmethod_prototype \\\n __unused SEL rxSelector = RX_selector(selector); \\\n IMP (^newImplementationGenerator)() = ^() { \\\n __block IMP thisIMP = nil; \\\n id newImplementation = ^return_value(__unsafe_unretained id self DECLARE_ARGUMENTS(__VA_ARGS__)) { \\\n body(__VA_ARGS__) \\\n \\\n struct objc_super superInfo = { \\\n .receiver = self, \\\n .super_class = class_getSuperclass(class) \\\n }; \\\n \\\n return_value (*msgSend)(struct objc_super *, SEL DECLARE_ARGUMENTS(__VA_ARGS__)) \\\n = (__typeof__(msgSend))objc_msgSendSuper; \\\n return msgSend(&superInfo, selector ARGUMENTS(__VA_ARGS__)); \\\n }; \\\n \\\n thisIMP = imp_implementationWithBlock(newImplementation); \\\n return thisIMP; \\\n }; \\\n \\\n IMP (^replacementImplementationGenerator)(IMP) = ^(IMP originalImplementation) { \\\n __block return_value (*originalImplementationTyped)(__unsafe_unretained id, SEL DECLARE_ARGUMENTS(__VA_ARGS__) ) \\\n = (__typeof__(originalImplementationTyped))(originalImplementation); \\\n \\\n __block IMP thisIMP = nil; \\\n id implementationReplacement = ^return_value(__unsafe_unretained id self DECLARE_ARGUMENTS(__VA_ARGS__) ) { \\\n body(__VA_ARGS__) \\\n \\\n return originalImplementationTyped(self, selector ARGUMENTS(__VA_ARGS__)); \\\n }; \\\n \\\n thisIMP = imp_implementationWithBlock(implementationReplacement); \\\n return thisIMP; \\\n }; \\\n \\\n return [self ensureSwizzledSelector:selector \\\n ofClass:class \\\n newImplementationGenerator:newImplementationGenerator \\\n replacementImplementationGenerator:replacementImplementationGenerator \\\n error:error]; \\\n } \\\n\n\n@interface RXObjCRuntime (InfrastructureMethods)\n@end\n\n// MARK: Infrastructure Methods\n\n@implementation RXObjCRuntime (InfrastructureMethods)\n\nSWIZZLE_INFRASTRUCTURE_METHOD(\n void,\n swizzleForwardInvocation,\n ,\n @selector(forwardInvocation:),\n FORWARD_BODY,\n NSInvocationRef\n)\nSWIZZLE_INFRASTRUCTURE_METHOD(\n BOOL,\n swizzleRespondsToSelector,\n ,\n @selector(respondsToSelector:),\n RESPONDS_TO_SELECTOR_BODY,\n SEL\n)\nSWIZZLE_INFRASTRUCTURE_METHOD(\n Class __nonnull,\n swizzleClass,\n toActAs:(Class)actAsClass,\n @selector(class),\n CLASS_BODY\n)\nSWIZZLE_INFRASTRUCTURE_METHOD(\n NSMethodSignatureRef,\n swizzleMethodSignatureForSelector,\n ,\n @selector(methodSignatureForSelector:),\n METHOD_SIGNATURE_FOR_SELECTOR_BODY,\n SEL\n)\nSWIZZLE_INFRASTRUCTURE_METHOD(\n void,\n swizzleDeallocating,\n ,\n deallocSelector,\n DEALLOCATING_BODY\n)\n\n@end\n\n// MARK: Optimized intercepting methods for specific combination of parameter types\n\nSWIZZLE_OBSERVE_METHOD(void)\n\nSWIZZLE_OBSERVE_METHOD(void, id)\nSWIZZLE_OBSERVE_METHOD(void, char)\nSWIZZLE_OBSERVE_METHOD(void, short)\nSWIZZLE_OBSERVE_METHOD(void, int)\nSWIZZLE_OBSERVE_METHOD(void, long)\nSWIZZLE_OBSERVE_METHOD(void, rx_uchar)\nSWIZZLE_OBSERVE_METHOD(void, rx_ushort)\nSWIZZLE_OBSERVE_METHOD(void, rx_uint)\nSWIZZLE_OBSERVE_METHOD(void, rx_ulong)\nSWIZZLE_OBSERVE_METHOD(void, rx_block)\nSWIZZLE_OBSERVE_METHOD(void, float)\nSWIZZLE_OBSERVE_METHOD(void, double)\nSWIZZLE_OBSERVE_METHOD(void, SEL)\n\nSWIZZLE_OBSERVE_METHOD(void, id, id)\nSWIZZLE_OBSERVE_METHOD(void, id, char)\nSWIZZLE_OBSERVE_METHOD(void, id, short)\nSWIZZLE_OBSERVE_METHOD(void, id, int)\nSWIZZLE_OBSERVE_METHOD(void, id, long)\nSWIZZLE_OBSERVE_METHOD(void, id, rx_uchar)\nSWIZZLE_OBSERVE_METHOD(void, id, rx_ushort)\nSWIZZLE_OBSERVE_METHOD(void, id, rx_uint)\nSWIZZLE_OBSERVE_METHOD(void, id, rx_ulong)\nSWIZZLE_OBSERVE_METHOD(void, id, rx_block)\nSWIZZLE_OBSERVE_METHOD(void, id, float)\nSWIZZLE_OBSERVE_METHOD(void, id, double)\nSWIZZLE_OBSERVE_METHOD(void, id, SEL)\n\n// MARK: RXObjCRuntime\n\n@implementation RXObjCRuntime\n\nstatic RXObjCRuntime *_instance = nil;\nstatic NSMutableDictionary<NSString *, RXInterceptWithOptimizedObserver> *optimizedObserversByMethodEncoding = nil;\n\n+(RXObjCRuntime*)instance {\n return _instance;\n}\n\n+(void)initialize {\n _instance = [[RXObjCRuntime alloc] init];\n defaultTypeID = CFGetTypeID((CFTypeRef)RXObjCRuntime.class); // just need a reference of some object not from CF\n deallocSelector = NSSelectorFromString(@\"dealloc\");\n NSAssert(_instance !", "= nil, @\"Failed to initialize swizzling\");\n}\n\n-(instancetype)init {\n self = [super init];\n if (!", "self) return nil;\n\n self.classesThatSupportObservingByForwarding = [NSMutableSet set];\n self.forwardedSelectorsByClass = [NSMutableDictionary dictionary];\n\n self.dynamicSubclassByRealClass = [NSMutableDictionary dictionary];\n self.interceptorIMPbySelectorsByClass = [NSMutableDictionary dictionary];\n\n pthread_mutexattr_t lock_attr;\n pthread_mutexattr_init(&lock_attr);\n pthread_mutexattr_settype(&lock_attr, PTHREAD_MUTEX_RECURSIVE);\n pthread_mutex_init(&_lock, &lock_attr);\n pthread_mutexattr_destroy(&lock_attr);\n \n return self;\n}\n\n-(void)performLocked:(void (^)(RXObjCRuntime* __nonnull))action {\n pthread_mutex_lock(&_lock);\n action(self);\n pthread_mutex_unlock(&_lock);\n}\n\n+(void)registerOptimizedObserver:(RXInterceptWithOptimizedObserver)registration encodedAs:(SEL)selector {\n Method exampleEncodingMethod = class_getClassMethod(self, selector);\n ALWAYS(exampleEncodingMethod !", "= nil, @\"Example encoding method is nil.\");", "\n\n NSString *methodEncoding = RX_method_encoding(exampleEncodingMethod);\n\n if (optimizedObserversByMethodEncoding == nil) {\n optimizedObserversByMethodEncoding = [NSMutableDictionary dictionary];\n }\n\n DLOG(@\"Added optimized method: %@ (%@)\", methodEncoding, NSStringFromSelector(selector));\n ALWAYS(optimizedObserversByMethodEncoding[methodEncoding] == nil, @\"Optimized observer already registered\")\n optimizedObserversByMethodEncoding[methodEncoding] = registration;\n}\n\n/**\n This is the main entry point for observing messages sent to arbitrary objects.", "\n */\n-(IMP __nullable)ensurePrepared:(id __nonnull)target forObserving:(SEL __nonnull)selector error:(NSError** __nonnull)error {\n Method instanceMethod = class_getInstanceMethod([target class], selector);\n if (instanceMethod == nil) {\n *error = [NSError errorWithDomain:RXObjCRuntimeErrorDomain\n code:RXObjCRuntimeErrorSelectorNotImplemented\n userInfo:nil];\n return nil;\n }\n\n if (selector == @selector(class)\n || selector == @selector(forwardingTargetForSelector:)\n || selector == @selector(methodSignatureForSelector:)\n || selector == @selector(respondsToSelector:)) {\n *error = [NSError errorWithDomain:RXObjCRuntimeErrorDomain\n code:RXObjCRuntimeErrorObservingPerformanceSensitiveMessages\n userInfo:nil];\n return nil;\n }\n\n // For `dealloc` message, original implementation will be swizzled.", "\n // This is a special case because observing `dealloc` message is performed when `rx_observeWeakly` is used.", "\n //\n // Some toll free bridged classes don't handle `object_setClass` well and cause crashes.", "\n //\n // To make `rx_deallocating` as robust as possible, original implementation will be replaced.", "\n if (selector == deallocSelector) {\n Class __nonnull deallocSwizzingTarget = [target class];\n IMP interceptorIMPForSelector = [self interceptorImplementationForSelector:selector forClass:deallocSwizzingTarget];\n if (interceptorIMPForSelector !", "= nil) {\n return interceptorIMPForSelector;\n }\n\n if (![", "self swizzleDeallocating:deallocSwizzingTarget error:error]) {\n return nil;\n }\n\n interceptorIMPForSelector = [self interceptorImplementationForSelector:selector forClass:deallocSwizzingTarget];\n if (interceptorIMPForSelector !", "= nil) {\n return interceptorIMPForSelector;\n }\n }\n else {\n Class __nullable swizzlingImplementorClass = [self prepareTargetClassForObserving:target error:error];\n if (swizzlingImplementorClass == nil) {\n return nil;\n }\n\n NSString *methodEncoding = RX_method_encoding(instanceMethod);\n RXInterceptWithOptimizedObserver optimizedIntercept = optimizedObserversByMethodEncoding[methodEncoding];\n\n if (!", "RX_method_has_supported_return_type(instanceMethod)) {\n *error = [NSError errorWithDomain:RXObjCRuntimeErrorDomain\n code:RXObjCRuntimeErrorObservingMessagesWithUnsupportedReturnType\n userInfo:nil];\n\n return nil;\n }\n\n // optimized interception method\n if (optimizedIntercept !", "= nil) {\n IMP interceptorIMPForSelector = [self interceptorImplementationForSelector:selector forClass:swizzlingImplementorClass];\n if (interceptorIMPForSelector !", "= nil) {\n return interceptorIMPForSelector;\n }\n\n if (!", "optimizedIntercept(self, swizzlingImplementorClass, selector, error)) {\n return nil;\n }\n\n interceptorIMPForSelector = [self interceptorImplementationForSelector:selector forClass:swizzlingImplementorClass];\n if (interceptorIMPForSelector !", "= nil) {\n return interceptorIMPForSelector;\n }\n }\n // default fallback to observing by forwarding messages\n else {\n if ([self forwardingSelector:selector forClass:swizzlingImplementorClass]) {\n return RX_default_target_implementation();\n }\n\n if (![", "self observeByForwardingMessages:swizzlingImplementorClass\n selector:selector\n target:target\n error:error]) {\n return nil;\n }\n\n if ([self forwardingSelector:selector forClass:swizzlingImplementorClass]) {\n return RX_default_target_implementation();\n }\n }\n }\n\n\n *error = [NSError errorWithDomain:RXObjCRuntimeErrorDomain\n code:RXObjCRuntimeErrorUnknown\n userInfo:nil];\n\n return nil;\n}\n\n-(Class __nullable)prepareTargetClassForObserving:(id __nonnull)target error:(NSError **__nonnull)error {\n Class swizzlingClass = objc_getAssociatedObject(target, &RxSwizzlingTargetClassKey);\n if (swizzlingClass !", "= nil) {\n return swizzlingClass;\n }\n\n Class __nonnull wannaBeClass = [target class];\n /**\n Core Foundation classes are usually toll free bridged. ", "Those classes crash the program in case\n `object_setClass` is performed on them.", "\n\n There is a possibility to just swizzle methods on original object, but since those won't be usual use\n cases for this library, then an error will just be reported for now.", "\n */\n BOOL isThisTollFreeFoundationClass = CFGetTypeID((CFTypeRef)target) !", "= defaultTypeID;\n\n if (isThisTollFreeFoundationClass) {\n *error = [NSError errorWithDomain:RXObjCRuntimeErrorDomain\n code:RXObjCRuntimeErrorCantInterceptCoreFoundationTollFreeBridgedObjects\n userInfo:nil];\n return nil;\n }\n\n /**\n If the object is reporting a different class then what it's real class, that means that there is probably\n already some interception mechanism in place or something weird is happening.", "\n \n Most common case when this would happen is when using KVO (`rx_observe`) and `rx_sentMessage`.", "\n\n This error is easily resolved by just using `rx_sentMessage` observing before `rx_observe`.", "\n \n The reason why other way around could create issues is because KVO will unregister it's interceptor \n class and restore original class. ", "Unfortunately that will happen no matter was there another interceptor\n subclass registered in hierarchy or not.", "\n \n Failure scenario:\n * KVO sets class to be `__KVO__OriginalClass` (subclass of `OriginalClass`)\n * `rx_sentMessage` sets object class to be `_RX_namespace___KVO__OriginalClass` (subclass of `__KVO__OriginalClass`)\n * then unobserving with KVO will restore class to be `OriginalClass` -> failure point\n\n The reason why changing order of observing works is because any interception method should return\n object's original real class (if that doesn't happen then it's really easy to argue that's a bug\n in that other library).", "\n \n This library won't remove registered interceptor even if there aren't any observers left because\n it's highly unlikely it would have any benefit in real world use cases, and it's even more\n dangerous.", "\n */\n if ([target class] !", "= object_getClass(target)) {\n BOOL isKVO = [target respondsToSelector:NSSelectorFromString(@\"_isKVOA\")];\n\n *error = [NSError errorWithDomain:RXObjCRuntimeErrorDomain\n code:RXObjCRuntimeErrorObjectMessagesAlreadyBeingIntercepted\n userInfo:@{\n RXObjCRuntimeErrorIsKVOKey: @(isKVO)\n }];\n return nil;\n }\n\n Class __nullable dynamicFakeSubclass = [self ensureHasDynamicFakeSubclass:wannaBeClass error:error];\n\n if (dynamicFakeSubclass == nil) {\n return nil;\n }\n\n Class previousClass = object_setClass(target, dynamicFakeSubclass);\n if (previousClass !", "= wannaBeClass) {\n *error = [NSError errorWithDomain:RXObjCRuntimeErrorDomain\n code:RXObjCRuntimeErrorThreadingCollisionWithOtherInterceptionMechanism\n userInfo:nil];\n THREADING_HAZARD(wannaBeClass);\n return nil;\n }\n\n objc_setAssociatedObject(target, &RxSwizzlingTargetClassKey, dynamicFakeSubclass, OBJC_ASSOCIATION_RETAIN_NONATOMIC);\n return dynamicFakeSubclass;\n}\n\n\n-(BOOL)forwardingSelector:(SEL)selector forClass:(Class __nonnull)class {\n return [self.forwardedSelectorsByClass[CLASS_VALUE(class)] containsObject:SEL_VALUE(selector)];\n}\n\n-(void)registerForwardedSelector:(SEL)selector forClass:(Class __nonnull)class {\n NSValue *classValue = CLASS_VALUE(class);\n\n NSMutableSet<NSValue *> *forwardedSelectors = self.forwardedSelectorsByClass[classValue];\n\n if (forwardedSelectors == nil) {\n forwardedSelectors = [NSMutableSet set];\n self.forwardedSelectorsByClass[classValue] = forwardedSelectors;\n }\n\n [forwardedSelectors addObject:SEL_VALUE(selector)];\n}\n\n-(BOOL)observeByForwardingMessages:(Class __nonnull)swizzlingImplementorClass\n selector:(SEL)selector\n target:(id __nonnull)target\n error:(NSError **__nonnull)error {\n if (![", "self ensureForwardingMethodsAreSwizzled:swizzlingImplementorClass error:error]) {\n return NO;\n }\n\n ALWAYS(![self forwardingSelector:selector forClass:swizzlingImplementorClass], @\"Already observing selector for class\");\n\n#if TRACE_RESOURCES\n OSAtomicIncrement32(&numberOfForwardedMethods);\n#endif\n SEL rxSelector = RX_selector(selector);\n\n Method instanceMethod = class_getInstanceMethod(swizzlingImplementorClass, selector);\n ALWAYS(instanceMethod !", "= nil, @\"Instance method is nil\");\n\n const char* methodEncoding = method_getTypeEncoding(instanceMethod);\n ALWAYS(methodEncoding !", "= nil, @\"Method encoding is nil.\");", "\n NSMethodSignature *methodSignature = [NSMethodSignature signatureWithObjCTypes:methodEncoding];\n ALWAYS(methodSignature !", "= nil, @\"Method signature is invalid.\");", "\n\n IMP implementation = method_getImplementation(instanceMethod);\n\n if (implementation == nil) {\n *error = [NSError errorWithDomain:RXObjCRuntimeErrorDomain\n code:RXObjCRuntimeErrorSelectorNotImplemented\n userInfo:nil];\n\n return NO;\n }\n\n if (!", "class_addMethod(swizzlingImplementorClass, rxSelector, implementation, methodEncoding)) {\n *error = [NSError errorWithDomain:RXObjCRuntimeErrorDomain\n code:RXObjCRuntimeErrorSavingOriginalForwardingMethodFailed\n userInfo:nil];\n return NO;\n }\n\n if (!", "class_addMethod(swizzlingImplementorClass, selector, _objc_msgForward, methodEncoding)) {\n if (implementation !", "= method_setImplementation(instanceMethod, _objc_msgForward)) {\n *error = [NSError errorWithDomain:RXObjCRuntimeErrorDomain\n code:RXObjCRuntimeErrorReplacingMethodWithForwardingImplementation\n userInfo:nil];\n THREADING_HAZARD(swizzlingImplementorClass);\n return NO;\n }\n }\n\n DLOG(@\"Rx uses forwarding to observe `%@` for `%@`.\", ", "NSStringFromSelector(selector), swizzlingImplementorClass);\n [self registerForwardedSelector:selector forClass:swizzlingImplementorClass];\n\n return YES;\n}\n\n/**\n If object don't have some weird behavior, claims it's the same class that runtime shows,\n then dynamic subclass is created (only this instance will have performance hit).", "\n \n In case something weird is detected, then original base class is being swizzled and all instances\n will have somewhat reduced performance.", "\n \n This is especially handy optimization for weak KVO. ", "Nobody will swizzle for example `NSString`,\n but to know when instance of a `NSString` was deallocated, performance hit will be only felt on a \n single instance of `NSString`, not all instances of `NSString`s.", "\n */\n-(Class __nullable)ensureHasDynamicFakeSubclass:(Class __nonnull)class error:(NSError **)error {\n Class dynamicFakeSubclass = self.dynamicSubclassByRealClass[CLASS_VALUE(class)];\n if (dynamicFakeSubclass !", "= nil) {\n return dynamicFakeSubclass;\n }\n\n NSString *dynamicFakeSubclassName = [RX_PREFIX stringByAppendingString:NSStringFromClass(class)];\n const char *dynamicFakeSubclassNameRaw = dynamicFakeSubclassName.", "UTF8String;\n dynamicFakeSubclass = objc_allocateClassPair(class, dynamicFakeSubclassNameRaw, 0);\n ALWAYS(dynamicFakeSubclass !", "= nil, @\"Class not generated\");\n\n if (![", "self swizzleClass:dynamicFakeSubclass toActAs:class error:error]) {\n return nil;\n }\n\n objc_registerClassPair(dynamicFakeSubclass);\n\n [self.dynamicSubclassByRealClass setObject:dynamicFakeSubclass forKey:CLASS_VALUE(class)];\n ALWAYS(self.dynamicSubclassByRealClass[CLASS_VALUE(class)] !", "= nil, @\"Class not registered\");\n\n return dynamicFakeSubclass;\n}\n\n-(BOOL)ensureForwardingMethodsAreSwizzled:(Class __nonnull)class error:(NSError ** __nonnull)error {\n NSValue *classValue = CLASS_VALUE(class);\n if ([self.classesThatSupportObservingByForwarding containsObject:classValue]) {\n return YES;\n }\n\n if (![", "self swizzleForwardInvocation:class error:error]) { return NO; }\n if (![", "self swizzleMethodSignatureForSelector:class error:error]) { return NO; }\n if (![", "self swizzleRespondsToSelector:class error:error]) { return NO; }\n\n [self.classesThatSupportObservingByForwarding addObject:classValue];\n\n return YES;\n}\n\n-(void)registerInterceptedSelector:(SEL)selector implementation:(IMP)implementation forClass:(Class)class {\n NSValue * __nonnull classValue = CLASS_VALUE(class);\n NSValue * __nonnull selectorValue = SEL_VALUE(selector);\n\n NSMutableDictionary *swizzledIMPBySelectorsForClass = self.interceptorIMPbySelectorsByClass[classValue];\n\n if (swizzledIMPBySelectorsForClass == nil) {\n swizzledIMPBySelectorsForClass = [NSMutableDictionary dictionary];\n self.interceptorIMPbySelectorsByClass[classValue] = swizzledIMPBySelectorsForClass;\n }\n\n swizzledIMPBySelectorsForClass[selectorValue] = IMP_VALUE(implementation);\n\n ALWAYS([self interceptorImplementationForSelector:selector forClass:class] !", "= nil, @\"Class should have been swizzled\");\n}\n\n-(IMP)interceptorImplementationForSelector:(SEL)selector forClass:(Class)class {\n NSValue * __nonnull classValue = CLASS_VALUE(class);\n NSValue * __nonnull selectorValue = SEL_VALUE(selector);\n\n NSMutableDictionary *swizzledIMPBySelectorForClass = self.interceptorIMPbySelectorsByClass[classValue];\n\n NSValue *impValue = swizzledIMPBySelectorForClass[selectorValue];\n return impValue.pointerValue;\n}\n\n-(BOOL)ensureSwizzledSelector:(SEL __nonnull)selector\n ofClass:(Class __nonnull)class\n newImplementationGenerator:(IMP(^)())newImplementationGenerator\nreplacementImplementationGenerator:(IMP (^)(IMP originalImplementation))replacementImplementationGenerator\n error:(NSError ** __nonnull)error {\n if ([self interceptorImplementationForSelector:selector forClass:class] !", "= nil) {\n DLOG(@\"Trying to register same intercept at least once, this sounds like a possible bug\");\n return YES;\n }\n\n#if TRACE_RESOURCES\n OSAtomicIncrement32(&numberOInterceptedMethods);\n#endif\n \n DLOG(@\"Rx is swizzling `%@` for `%@`\", NSStringFromSelector(selector), class);\n\n Method existingMethod = class_getInstanceMethod(class, selector);\n ALWAYS(existingMethod !", "= nil, @\"Method doesn't exist\");\n\n const char *encoding = method_getTypeEncoding(existingMethod);\n ALWAYS(encoding !", "= nil, @\"Encoding is nil\");\n\n IMP newImplementation = newImplementationGenerator();\n\n if (class_addMethod(class, selector, newImplementation, encoding)) {\n // new method added, job done\n [self registerInterceptedSelector:selector implementation:newImplementation forClass:class];\n\n return YES;\n }\n\n imp_removeBlock(newImplementation);\n\n // if add fails, that means that method already exists on targetClass\n Method existingMethodOnTargetClass = existingMethod;\n\n IMP originalImplementation = method_getImplementation(existingMethodOnTargetClass);\n ALWAYS(originalImplementation !", "= nil, @\"Method must exist.\");", "\n IMP implementationReplacementIMP = replacementImplementationGenerator(originalImplementation);\n ALWAYS(implementationReplacementIMP !", "= nil, @\"Method must exist.\");", "\n IMP originalImplementationAfterChange = method_setImplementation(existingMethodOnTargetClass, implementationReplacementIMP);\n ALWAYS(originalImplementation !", "= nil, @\"Method must exist.\");", "\n\n // If method replacing failed, who knows what happened, better not trying again, otherwise program can get\n // corrupted.", "\n [self registerInterceptedSelector:selector implementation:implementationReplacementIMP forClass:class];\n\n // ¯\\_(ツ)_/¯\n if (originalImplementationAfterChange !", "= originalImplementation) {\n THREADING_HAZARD(class);\n return NO;\n }\n\n return YES;\n}\n\n@end\n\n#if TRACE_RESOURCES\n\nNSInteger RX_number_of_dynamic_subclasses() {\n __block NSInteger count = 0;\n [[RXObjCRuntime instance] performLocked:^(RXObjCRuntime * __nonnull self) {\n count = self.dynamicSubclassByRealClass.count;\n }];\n\n return count;\n}\n\nNSInteger RX_number_of_forwarding_enabled_classes() {\n __block NSInteger count = 0;\n [[RXObjCRuntime instance] performLocked:^(RXObjCRuntime * __nonnull self) {\n count = self.classesThatSupportObservingByForwarding.count;\n }];\n\n return count;\n}\n\nNSInteger RX_number_of_intercepting_classes() {\n __block NSInteger count = 0;\n [[RXObjCRuntime instance] performLocked:^(RXObjCRuntime * __nonnull self) {\n count = self.interceptorIMPbySelectorsByClass.count;\n }];\n\n return count;\n}\n\nNSInteger RX_number_of_forwarded_methods() {\n return numberOfForwardedMethods;\n}\n\nNSInteger RX_number_of_swizzled_methods() {\n return numberOInterceptedMethods;\n}\n\n#endif\n\n#endif" ]

{ "pile_set_name": "Github" }

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[ "/*\n * Linux driver for TerraTec DMX 6Fire USB\n *\n * Main routines and module definitions.", "\n *\n * Author:\tTorsten Schenk <torsten.schenk@zoho.com>\n * Created:\tJan 01, 2011\n * Copyright:\t(C) Torsten Schenk\n *\n * This program is free software; you can redistribute it and/or modify\n * it under the terms of the GNU General Public License as published by\n * the Free Software Foundation; either version 2 of the License, or\n * (at your option) any later version.", "\n */\n\n#include \"chip.h\"\n#include \"firmware.h\"\n#include \"pcm.h\"\n#include \"control.h\"\n#include \"comm.h\"\n#include \"midi.h\"\n\n#include <linux/moduleparam.h>\n#include <linux/interrupt.h>\n#include <linux/module.h>\n#include <linux/init.h>\n#include <linux/gfp.h>\n#include <sound/initval.h>\n\nMODULE_AUTHOR(\"Torsten Schenk <torsten.schenk@zoho.com>\");\nMODULE_DESCRIPTION(\"TerraTec DMX 6Fire USB audio driver\");\nMODULE_LICENSE(\"GPL v2\");\nMODULE_SUPPORTED_DEVICE(\"{{TerraTec,DMX 6Fire USB}}\");\n\nstatic int index[SNDRV_CARDS] = SNDRV_DEFAULT_IDX; /* Index 0-max */\nstatic char *id[SNDRV_CARDS] = SNDRV_DEFAULT_STR; /* Id for card */\nstatic bool enable[SNDRV_CARDS] = SNDRV_DEFAULT_ENABLE_PNP; /* Enable card */\nstatic struct sfire_chip *chips[SNDRV_CARDS] = SNDRV_DEFAULT_PTR;\nstatic struct usb_device *devices[SNDRV_CARDS] = SNDRV_DEFAULT_PTR;\n\nmodule_param_array(index, int, NULL, 0444);\nMODULE_PARM_DESC(index, \"Index value for the 6fire sound device\");\nmodule_param_array(id, charp, NULL, 0444);\nMODULE_PARM_DESC(id, \"ID string for the 6fire sound device.\");", "\nmodule_param_array(enable, bool, NULL, 0444);\nMODULE_PARM_DESC(enable, \"Enable the 6fire sound device.\");", "\n\nstatic DEFINE_MUTEX(register_mutex);\n\nstatic void usb6fire_chip_abort(struct sfire_chip *chip)\n{\n\tif (chip) {\n\t\tif (chip->pcm)\n\t\t\tusb6fire_pcm_abort(chip);\n\t\tif (chip->midi)\n\t\t\tusb6fire_midi_abort(chip);\n\t\tif (chip->comm)\n\t\t\tusb6fire_comm_abort(chip);\n\t\tif (chip->control)\n\t\t\tusb6fire_control_abort(chip);\n\t\tif (chip->card) {\n\t\t\tsnd_card_disconnect(chip->card);\n\t\t\tsnd_card_free_when_closed(chip->card);\n\t\t\tchip->card = NULL;\n\t\t}\n\t}\n}\n\nstatic void usb6fire_chip_destroy(struct sfire_chip *chip)\n{\n\tif (chip) {\n\t\tif (chip->pcm)\n\t\t\tusb6fire_pcm_destroy(chip);\n\t\tif (chip->midi)\n\t\t\tusb6fire_midi_destroy(chip);\n\t\tif (chip->comm)\n\t\t\tusb6fire_comm_destroy(chip);\n\t\tif (chip->control)\n\t\t\tusb6fire_control_destroy(chip);\n\t\tif (chip->card)\n\t\t\tsnd_card_free(chip->card);\n\t}\n}\n\nstatic int usb6fire_chip_probe(struct usb_interface *intf,\n\t\t\t const struct usb_device_id *usb_id)\n{\n\tint ret;\n\tint i;\n\tstruct sfire_chip *chip = NULL;\n\tstruct usb_device *device = interface_to_usbdev(intf);\n\tint regidx = -1; /* index in module parameter array */\n\tstruct snd_card *card = NULL;\n\n\t/* look if we already serve this card and return if so */\n\tmutex_lock(&register_mutex);\n\tfor (i = 0; i < SNDRV_CARDS; i++) {\n\t\tif (devices[i] == device) {\n\t\t\tif (chips[i])\n\t\t\t\tchips[i]->intf_count++;\n\t\t\tusb_set_intfdata(intf, chips[i]);\n\t\t\tmutex_unlock(&register_mutex);\n\t\t\treturn 0;\n\t\t} else if (!", "devices[i] && regidx < 0)\n\t\t\tregidx = i;\n\t}\n\tif (regidx < 0) {\n\t\tmutex_unlock(&register_mutex);\n\t\tdev_err(&intf->dev, \"too many cards registered.\\n\");\n\t\treturn -ENODEV;\n\t}\n\tdevices[regidx] = device;\n\tmutex_unlock(&register_mutex);\n\n\t/* check, if firmware is present on device, upload it if not */\n\tret = usb6fire_fw_init(intf);\n\tif (ret < 0)\n\t\treturn ret;\n\telse if (ret == FW_NOT_READY) /* firmware update performed */\n\t\treturn 0;\n\n\t/* if we are here, card can be registered in alsa. */", "\n\tif (usb_set_interface(device, 0, 0) !", "= 0) {\n\t\tdev_err(&intf->dev, \"can't set first interface.\\n\");\n\t\treturn -EIO;\n\t}\n\tret = snd_card_new(&intf->dev, index[regidx], id[regidx],\n\t\t\t THIS_MODULE, sizeof(struct sfire_chip), &card);\n\tif (ret < 0) {\n\t\tdev_err(&intf->dev, \"cannot create alsa card.\\n\");\n\t\treturn ret;\n\t}\n\tstrcpy(card->driver, \"6FireUSB\");\n\tstrcpy(card->shortname, \"TerraTec DMX6FireUSB\");\n\tsprintf(card->longname, \"%s at %d:%d\", card->shortname,\n\t\t\tdevice->bus->busnum, device->devnum);\n\n\tchip = card->private_data;\n\tchips[regidx] = chip;\n\tchip->dev = device;\n\tchip->regidx = regidx;\n\tchip->intf_count = 1;\n\tchip->card = card;\n\n\tret = usb6fire_comm_init(chip);\n\tif (ret < 0) {\n\t\tusb6fire_chip_destroy(chip);\n\t\treturn ret;\n\t}\n\n\tret = usb6fire_midi_init(chip);\n\tif (ret < 0) {\n\t\tusb6fire_chip_destroy(chip);\n\t\treturn ret;\n\t}\n\n\tret = usb6fire_pcm_init(chip);\n\tif (ret < 0) {\n\t\tusb6fire_chip_destroy(chip);\n\t\treturn ret;\n\t}\n\n\tret = usb6fire_control_init(chip);\n\tif (ret < 0) {\n\t\tusb6fire_chip_destroy(chip);\n\t\treturn ret;\n\t}\n\n\tret = snd_card_register(card);\n\tif (ret < 0) {\n\t\tdev_err(&intf->dev, \"cannot register card.\");", "\n\t\tusb6fire_chip_destroy(chip);\n\t\treturn ret;\n\t}\n\tusb_set_intfdata(intf, chip);\n\treturn 0;\n}\n\nstatic void usb6fire_chip_disconnect(struct usb_interface *intf)\n{\n\tstruct sfire_chip *chip;\n\tstruct snd_card *card;\n\n\tchip = usb_get_intfdata(intf);\n\tif (chip) { /* if !", "chip, fw upload has been performed */\n\t\tcard = chip->card;\n\t\tchip->intf_count--;\n\t\tif (!", "chip->intf_count) {\n\t\t\tmutex_lock(&register_mutex);\n\t\t\tdevices[chip->regidx] = NULL;\n\t\t\tchips[chip->regidx] = NULL;\n\t\t\tmutex_unlock(&register_mutex);\n\n\t\t\tchip->shutdown = true;\n\t\t\tusb6fire_chip_abort(chip);\n\t\t\tusb6fire_chip_destroy(chip);\n\t\t}\n\t}\n}\n\nstatic struct usb_device_id device_table[] = {\n\t{\n\t\t.match_flags = USB_DEVICE_ID_MATCH_DEVICE,\n\t\t.idVendor = 0x0ccd,\n\t\t.idProduct = 0x0080\n\t},\n\t{}\n};\n\nMODULE_DEVICE_TABLE(usb, device_table);\n\nstatic struct usb_driver usb_driver = {\n\t.name = \"snd-usb-6fire\",\n\t.probe = usb6fire_chip_probe,\n\t.disconnect = usb6fire_chip_disconnect,\n\t.id_table = device_table,\n};\n\nmodule_usb_driver(usb_driver);\n" ]

{ "pile_set_name": "Github" }

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[ "George Will warns of ‘mission creep’ in Libya\n\nNot everyone on the right thinks that U.S. involvement in the operation against Libyan leader Muammar Gaddafi is a good idea.", "\n\nABC News invited Iraq war architect Paul Wolfowitz and conservative columnist George Will Sunday to discuss the attack on Libya. ", "Will seemed to have the Iraq war in mind when he warned Wolfowitz against “mission creep” in the Middle East.", "\n\n“Do you think this was the right thing to do?” ", "ABC’s Christian Amanpour asked Will.", "\n\n“I do not,” Will said. “", "We have intervened in a tribal society in a civil war. ", "And we’ve taken sides in that civil war on behalf of people we do not know or understand for the purpose of creating a political vacuum by decapitating that government. ", "Into that vacuum, what will flow? ", "We do not know. ", "We cannot know.”", "\n\nWolfowitz quickly disagreed.", "\n\n“I understand George’s hesitations,” Wolfowitz said. “", "If you follow George’s hesitations, you say, it’s better to keep this devil we know than getting in someone new. ", "I don’t think anybody new could be worse than the devil in Tripoli right now.”", "\n\n“Wouldn’t you say the hesitation, you can trace it write back to your operation in Iraq?” ", "Amanpour pressed Wolfowitz. “", "There was such a pendulum swing against trying to intervene because of the chaos that was unleashed.”", "\n\n“We’ve paid the price of intervention sometimes,” Wolfowitz admitted. “", "We’ve paid the price of non-intervention, in Bosnia, for example.”", "\n\nAmanpour noted that there seemed to a “double standard” when it came to taking military action against Libya, but ignoring the regimes in Bahrain and Yemen.", "\n\n“You can’t compare the regime in Bahrain or even the regime in [Yemen capital city] Sanaa to Gaddafi,” Wolfowitz argued. “", "Yes, there is a certain — there’s something in common here, which is regimes that don’t represent their people, they’re not only wrong their ultimately unstable.”", "\n\n“I think what we should be working for in Bahrain, what we should be working for in Yemen are governments that are much more representative of their people so we can work with them better. ", "But they’re not — it’s absolutely wrong to compare what’s happening there to what Gaddafi is doing and has been doing for 40 years,” he added.", "\n\n“There is no limiting principle in what we’ve done,” Will countered. “", "If we are to protect people under assault, then where people are under assault in Bahrain, we’re logically committed to help them. ", "We’re inciting them to rise up in expectation.”", "\n\n“The mission creep here began, Paul, before the mission began,” he told Wolfowitz. “", "Because we had a means not suited to the end. ", "The means is a no-fly zone. ", "That will not affect the end, which is obviously regime change.”", "\n\nAbout the Author\n\nDavid Edwards has served as an editor at Raw Story since 2006. ", "His work can also be found at Crooks & Liars, and he's also been published at The BRAD BLOG. ", "He came to Raw Story after working as a network manager for the state of North Carolina and as as engineer developing enterprise resource planning software. ", "Follow him on Twitter at @DavidEdwards." ]

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[ "Product Reviews\n\nStops all squirrels!, ", "February 22, 2013\n\nPosted By:John - Verified BuyerLocation: Cape Cod Massachusetts How often do you shop at Comfort House: first time\n\nThis is the greatest. ", "We have tried all the so-called squirrel prooffeeders on the market and this is the best. ", "My spouse and I watch in amazement as squirrels attempt to do their thing with this feeder and are never successful. ", "The birds definitely love it.", "\n\nperfect, January 19, 2012\n\nPosted By:arlene - Verified BuyerLocation: pa How often do you shop at Comfort House: first time\n\nSquirrels fall off as soon as they touch the baffle. ", "Very well made product that was easy to assemble.", "\n\nNO MORE SQUIRRELS !!, ", "November 10, 2011\n\nPosted By:Randy - Verified BuyerLocation: Baldwin How often do you shop at Comfort House: first time\n\nAMAZING!!! ", "This product did exactly what it said it does. ", "We recently moved to a rural area - squirrels were stealing all the bird food. ", "Frustrated - I resorted to stinging them with a BB gun. ( ", "figured there was a better way -- more humane / less time consuming) - Squirrels don't even try now!!!" ]

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[ "NOW PLAYING\n\nIran promises retaliation if attacked\n\nIran will retaliate to any attack against the Islamic republic by Israel or the US, a top Revolutionary Guards commander says.", "\n\n26 Jan 2005 07:56 GMT\n\nIran spends about $10 billion a year on its military\n\nBrigadier-General Muhammad Ali Jafari told the Shargh newspaper on Wednesday that Tehran \"will counter any stupid action by Israel and its master with firmness and in an astonishing way\".", "\n\nThe commander of the Revolutionary Guards' ground forces said Iran now had the capacity to defeat any invader within months.", "\n\n\"We pushed the Baathist enemy from our country within one and a half years,\" he said, referring to the opening campaigns of the 1980-88 war with Iraq.", "\n\n\"With the experience and skills from that war and in the case of any invasion, the invaders will be defeated in less than one and a half months,\" Jafari said.", "\n\nUS officials have in recent days hardened their tone against the Islamic republic, which US President George Bush had earlier lumped into an \"axis of evil\" with Iraq and North Korea.", "\n\nIncreasing tensions\n\nLast Monday, Bush said he could not rule out using force if Tehran failed to rein in its nuclear plans, and US Vice-President Dick Cheney said Iran was \"right at the top of the list\" of global trouble spots.", "\n\n\"We would love American commandos to enter Iran ... because they are chicks. ", "Our eagles will catch them in no time. ", "The Americans are stupid, but not as stupid as that\"\n\nAli Yunusi,\n\nIranian intelligence minister\n\nCheney also warned that Israel might launch a pre-emptive strike on its own to shut down Iran's nuclear programme.", "\n\nBut Iranian Intelligence Minister Ali Yunusi responded to both Bush and Cheney's remarks in a broadcast on Iran's Channel two.", "\n\n\"This is a psychological war that they have started in order to influence the talks on the nuclear issue.", "\n\n\"They think that this way they can force us to make the concessions they want. ", "Otherwise, they do not have the ability to carry out their military threat,\" he told Aljazeera.net.", "\n\nWhen asked if he believed claims made by US journalist Seymour Hersh that US commandos were operating inside Iran, Yunusi rejected the idea.", "\n\n\"We would love American commandos to enter Iran ... because they are chicks. ", "Our eagles will catch them in no time. ", "The Americans are stupid, but not as stupid as that.\"" ]

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[ "Related Articles\n\nGrouse breed in the wild, so numbers cannot be augmented as easily as pheasants and partridge, most of which are reared and then released by gamekeepers to be hunted.", "\n\nRobert Benson, the chairman of the Moorland Association, said pre-season counts of grouse in England showed that numbers were up.", "\n\n“Generally, at long last, for the first time in the last few years, we’ve had a reasonable nesting and hatching period, with good chick survival.”", "\n\nCKD Galbraith, the rural property and estates agency, said that in Scotland, where it has been dry and warm most of the ­summer, it could be “one of the best seasons for many years”.", "\n\nSome moorland managers have delayed the start of their shoots because of the late fledging of the birds, but this means shooting could carry on later into the year.", "\n\nThe chance of a plentiful season led to animal-welfare campaigners claiming that grouse-shooting “turns the picturesque moorlands into bloody killing fields”.", "\n\nThe RSPB is concerned that gamekeepers illegally kill rare birds of prey to protect grouse.", "\n\nFigures released in the past few days showed that no hen harriers successfully nested in England, pushing the bird of prey to the brink of extinction in the country. ", "However, the Game and Wildlife Conservation Trust recently published data showing that gamekeepers can help ­populations of both birds of prey and rare wading birds to thrive by controlling foxes and crows.", "\n\nThe warm weather has also been ripening plenty of berries, meaning wildlife should be able to fatten up for the winter, and keen foragers can stock up on blackberries.", "\n\nDr Kate Lewthwaite, of the Woodland Trust, who manages a public survey known as Nature’s Calendar to gauge the effect of climate change on wildlife, said that already people were noticing lot of berries growing.", "\n\n“Although our records suggest that autumn fruiting will be late this year due to the delayed onset of spring flowering, if the warm weather interspersed with occasional wet spells continues this should mean the fruiting of shrubs like bramble, rowan and blackthorn is abundant,” she said. “", "Wildlife species will no doubt benefit from a bumper crop, and finally fruit-eating birds and mammals will be able to enjoy an autumn feast.”", "\n\nFor the early part of the week temperatures are forecast to remain around 71.6F (22C), with thundery showers, rain and sunny spells until Wednesday. ", "There could be localised flooding and transport disruption caused by the downpours.", "\n\nHowever, on Thursday, warm air is due to arrive from the south, with temperatures rising back up to 78.8F (26C), or higher in the South East in the sunny spells between cloudy periods." ]

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[ "KEN: I was attracted to this project having had no prior knowledge of e-lit or digital humanities in the academic context. ", "I had taken a class with Jesse before, and I had really enjoyed it. ", "This was an opportunity to work with him and explore some of this new frontier.", "\n\nJESSE: At the start, I immediately thought, “Who is going to be the outlier?” ", "Who is going to be the person to push us in ways that we didn’t expect, and Ken was the best outlier I knew. ", "The other folks I invited to join had all been in my electronic literature class, so Ken was the one who was going to bring the outside perspective.", "\n\nLANS: I’m going to go off on a tangent. ", "My daughter went to Montessori school, which had a mixed-age environment. ", "Each class is made with a mentorship of older children and younger children, and the synergy between them is kind of like an internship. ", "I’d say the synergy is what attracted me to this project. ", "I like what you said about having somebody from outside that helps push and shape things in ways that are unexpected. ", "That’s my way of motivating myself to explore new possibilities in what I can do. ", "I tend to not really go there without a little bit of a push, and Jesse is really good at that.", "\n\nI was also excited that I knew it was Library of Congress. ", "There’s a lot of lit-bling attached to that. ", "I wanted to get that on my lapel.", "\n\nJESSICA: For me it was the prospect of working on a project in relation to the Library of Congress. ", "Because I’m a public relations minor, this project was a nice way for me to gain some of those skills in communication, social media and project management. ", "I already knew Jesse so I knew it was going to be a co-learning project rather than a “these are your tasks and you need to complete them by this deadline” situation. ", "It was a neat project because it was just so raw and flexible.", "\n\nJESSE: One of the things I loved about working with these folks is that there was a little bit of trepidation at the beginning about the disorganized quality of what we were doing. ", "We didn’t know where it was going, and you never know what’s going to happen with those kinds of collaborations. ", "But what happened is that each of us did things at the exact moment that we needed to. ", "People filled in gaps. ", "Nobody called someone else and asked, “Are you going to be tweeting today?” ", "Instead, each person recognized there was a gap and decided to fill it.", "\n\nKATHI: There was some interesting professional development in the project, and this part was led by Jesse. ", "He created a Twitter schedule. ", "He knows how to do a campaign launch. ", "He brought that expertise to bear, and it trained all of us. ", "And it was a huge launch. ", "I think we had 1,700 hits [at spinepoetry.com] in the first day.", "\n\nYou’ve used a lot of words to describe this process: raw, flexible, disorganized, natural. ", "You came to a decision about spine poetry. ", "I’m wondering how you came to that decision? ", "What did that decision-making process look like? ", "Why spine poetry?", "\n\nKEN: It felt like a pan-digital, academic Ouija board. ", "It was organizing itself through the collusion of us working together.", "\n\nKATHI: We wanted books because of the connection to the Library of Congress. ", "So we thought, What could possibly make a connection between the artifact of the book and what electronic literature does? ", "There was a moment when Carrie was filming and captured a shot of Ken, where Ken is holding a stack of books in the traditional way that you look at them. ", "He turned them horizontally, and that act of defamiliarizing it and saying, “Did you know that you can actually make poems out of these,” was the e-lit moment. ", "You start to read the container. ", "The container itself tells a story, and that’s foundational to e-lit. ", "And this was from the guy who was brand new to e-lit!", "\n\nJESSE: Lans had this idea at the beginning: How are we going to organize whatever it is that we create? ", "We didn’t even know what we would build. ", "She had this idea of organizing it around the Dewey decimal system. ", "We didn’t end up using that, but her idea ended up fueling this sense of “How can we take something chaotic and organize it?”", "\n\nCARRIE: The first thing we did was brainstorm. ", "And brainstorm. ", "And brainstorm. ", "And talk about what is cool in the digital? ", "What is happening in literature right now? ", "What interests us? ", "That’s where we started. ", "It all just happened to converge in the spine poetry direction.", "\n\nLANS: It was really spontaneous, natural and magnetic. ", "Even my daughter, who was in the library at the time we were discussing this, was drawn into our conversation from another table and took it upon herself—at fourteen—to make her own spine poems. ", "That sealed it for me. ", "It was very accessible, which is what electronic literature does for regularly inaccessible lit. ", "It opens it up.", "\n\nThat’s a perfect segue. ", "You’ve been talking about e-lit, and it was the e-lit showcase at the Library of Congress this past April. ", "What does e-lit mean for this group? ", "How would you describe it?", "\n\nJESSICA: That was part of the problem we were up against as a group. ", "We were working, the whole time, to define e-literature.", "\n\nCARRIE: It’s as slippery of an idea as “What is literature?” ", "because there are so many things that are electronic literature.", "\n\nKEN: You start to accrete characteristics more than hard definitions.", "\n\nCan you name some?", "\n\nKEN: The one that immediately comes to mind is digital storytelling.", "\n\nJESSE: I think it was Kathi who said this in our video: E-lit is literature that comes alive when you touch it. ", "It comes alive when you manipulate it, play with it. ", "It’s interactive. ", "It isn’t itself until it has a user.", "\n\nSo it’s not Google Books.", "\n\nKATHI: Exactly. ", "If you can print it out, it’s not e-lit. ", "If you can print it out and get the same thing you’d get from reading it on a tablet or phone, it’s not e-lit.", "\n\nJESSE: The reason spine poems are e-lit is not the artifact of the spine poem itself, but it’s the way it calls you to action to make your own.", "\n\nKATHI: And then the spreadability. ", "This is the other piece, too. ", "What makes a spreadable bit of content? ", "It has to be small. ", "Even the videos of people reading their spine poems are no more than ten seconds long. ", "So those are easy to share. ", "You can make your own without expertise. ", "A problem with a lot of e-lit—if you call it a problem—is that it can really require some expertise to read. ", "It can be off-putting because you have to have be willing to click around a bunch and see what happens. ", "If you’re a person who’s afraid it may break if you touch it, you are not really going to read e-lit.", "\n\nHow do you inculcate the sense that play can actually mean failure? ", "For many of us, play means “Yay, score! ", "You get a reward” as opposed to “Well, I tried that and that didn’t work. ", "Why would the author want me to do this?”", "\n\nLANS: It’s also so new and wild of an animal that we’re just discovering what potential it has. ", "We’re in the moment where we have to corral it. ", "The kill-or-be-killed moment. ", "Is everything going to go digital? ", "Are we going to reel it in? ", "How is that going to happen? ", "What can e-lit do? ", "Where does it go when copyright or websites disappear?", "\n\nGoing back to what you were saying about play, I remember during one of our first meetings, we asked each other, “What was your first experience with e-lit?” ", "And for me, it was The Oregon Trail game. ", "Or there was another Apple game in which I died because I got caught in a vacuum and couldn’t figure out how to get out in two words or less. ", "It was play.", "\n\nThis has already been discussed in this conversation, but why is e-literature—and this Library of Congress project—important?", "\n\nJESSICA: This process, for me, was really important because there is a lot of fear in going digital and the future of physical books. ", "I am one of those people. ", "It is a scary thing. ", "How do we move forward into the digital world but also keep the physical book?", "\n\nWhat became really important in this project was to show that digital literature is not a threat. ", "Whenever it’s talked about, it’s described as a replacement of “normal” literature. ", "But really it’s more of an enhancement of literature rather than a supplement.", "\n\nCARRIE: If we’re talking about academia, lately everything is so serious. ", "There is not enough play and experimentation. ", "Having a project where you invite people to touch something and figure it out and fail and try again was really important to the academic landscape.", "\n\nThis project was a course, correct?", "\n\nCARRIE: Yes.", "\n\nLANS: Another reason why I was drawn to be a part of this was because I could think of no better, more fun way to earn credits. ", "It was the kind of real experience that you get working on a real project you’d do after graduation. ", "That’s not something you typically get in a classroom unless you’re learning a skill.", "\n\nCARRIE: We weren’t just learning a skill. ", "We were learning how to figure out what skills we needed for the project. ", "There’s a lot of critical thinking in that.", "\n\nJESSICA: We learned so much from the launch experience. ", "It was a way of putting everything we had done into action. ", "We were reaching out to new communities and some of our own personal and professional circles. ", "We were able to learn by bringing people in and sharing our project with others.", "\n\nJESSE: We talked about this at one point. ", "We even put it on our homepage: we were going to promote your work. ", "That is such an important part of digital culture for me. ", "The generosity. ", "This is a culture of generosity. ", "That is the work that we need to be doing, which is boosting each other. ", "The reason we do it is because it’s part of the spreadability. ", "It’s almost like crowdsurfing. ", "You have to keep it moving. ", "Otherwise the person falls. ", "So when something is moving towards you [in the digital space], you have to boost it. ", "You have to share it.", "\n\nJESSICA: It was great to see people tweeting their stuff. ", "Even after the Library of Congress showcase was over, we’re still getting stuff in. ", "It’s great.", "\n\nLANS: We’re talking about what makes e-lit important, and I think it’s also important because it gives you good feelings when you’re behind the computer. ", "It’s something that makes you feel positive. ", "It’s so easy to be negative, so easy to share about hate crimes and war. ", "It’s sometimes hard to remember to be creative. ", "As easy as the digital makes it for us to multi-task, it does take our time away from being able to play, to have good feelings, to produce artwork. ", "E-lit gives you that moment. ", "It gives you a micro-break.", "\n\nWhat was the biggest surprise for each of you during this project?", "\n\nCARRIE: I think the biggest surprise for me was just how open-ended it was. ", "How much freedom we really had to determine what we were going to do. ", "I had fully expected to come in on the first day and have Kathi tell us what was happening. ", "That didn’t happen. ", "We talked around what we wanted to do, and there was some real fear and insecurity in that. ", "We didn’t know who was driving this crazy bus. ", "But it led to this beautiful, open, collaborative experience that you couldn’t have had any other way.", "\n\nLANS: It was surprising for me that while the project was pretty intense with quick deadlines, there was also a feeling of joy. ", "You don’t usually feel like “Whoo, I get to go to class today!” ", "To be able to look at my kid and say, “I get to go to class today,” and her knowing I’m going to go and have a great time, come back—fully energized—and produce things…that was not anything I had expected.", "\n\nJESSE: Something that did not surprise me at all was that I learned way more from these folks than I could have ever imagined teaching them.", "\n\nAt the end of this project, what gave you all the greatest sense of satisfaction or achievement?", "\n\nJESSICA: The initial launch was in itself a great achievement. ", "I felt a sense of achievement by seeing the project take off on its own.", "\n\nJESSE: When the students don’t need me anymore, that’s the payoff. ", "Same with this project. ", "We gave birth to the project, and it ran itself. ", "It became its own thing that we couldn’t control because we didn’t know what people were going to create.", "\n\nLANS: I’ve made connections on Twitter that I can’t even comprehend. ", "When I investigate them, I find that they are very respected in the field, and they know who I am. ", "We’re also affecting other local schools. ", "We reached out to high schools, and they’re having their classes do spine poetry.", "\n\nKEN: I loved how it enhanced my existing relationships with non-tech people. ", "I had all my coworkers and friends involved. ", "I still have people coming up to me saying, “I love this spine poetry thing. ", "I’m going to do some this weekend.”", "\n\nKATHI: That is so cool.", "\n\nJESSICA: My boyfriend just went to Wisconsin for his sister’s birthday. ", "They stopped by her university’s library, and he was showing them how to do it." ]

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[ "The effects of mycophenolate mofetil on encapsulated peritoneal sclerosis model in rats.", "\nEncapsulated peritoneal sclerosis (EPS) is a devastating complication of peritoneal dialysis. ", "We aimed to investigate the effects of mycophenolate mofetil (MMF) treatment in experimental EPS in rats. ", "40 nonuremic Wistar albino rats were divided equally into 4 groups: control rats received 2 ml isotonic saline intraperitoneally daily for 3 weeks without any other treatment. ", "The chlorhexidine gluconate group received intraperitoneally 2 ml/200 g injection of chlorhexidine gluconate and ethanol dissolved in saline for 3 weeks. ", "The resting group received chlorhexidine gluconate (0 - 3rd week) + peritoneal resting (4th - 6th week). ", "The MMF group received chlorhexidine gluconate (0 - 3rd week) + 125 mg/l MMF in drinking water (4th - 6th week). ", "Dialysate cytokine levels, leukocyte count, peritoneal thickness, inflammation and fibroblast activities were evaluated. ", "Although the MMF and resting groups showed beneficial effects on ultrafiltration and D1/D0 glucose compared to the chlorhexidine gluconate group, only MMF treatment improved dialysate TGFβ1, VEGF and MCP-1 levels compared to the resting group. ", "Inflammatory activity and vascularity observed in a tissue biopsy, including capillaries number per mm2 of submesothelial area, decreased in the treatment group. ", "MMF treatment has beneficial effects on EPS via inhibiting inflammation and neovascularisation by reducing dialysate VEGF overexpression." ]

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[ "click to enlarge Facebook / Geekdom SA\n\nand\n\ncybersecurity\n\nAustin may be considered the Silicon Valley of Texas, but San Antonio has a lot to offer to the tech industry, too.", "In its November 2018 issue, Inc. Magazine argued just that, saying that the Alamo City is a \"tech hotspot.\" ", "The magazine writes that San Antonio \"dominates cybersecurity,\" and has since 1985. ", "That year, the Air Force consolidated its electronic security mission to Military City USA. ", "That move brought defense contractors and eventually a National Security Agency outpost.", "Today, there's plenty of cybersecurity startups popping up throughout San Antonio. ", "So why isn't the 2-1-0 getting recognition for its place in the tech industry? ", "Inc. writes that the big companies here \"work in the shadows.\" ", "Companies like ThreatGuard, SecureLogix, SilotechInnové do important work that you might hear about on the news, but they're just not household names.", "Higher education across the city is helping propel San Antonio's place in the industry. ", "UTSA has three cybersecurity centers, one of them being the nation's top-ranked program. ", "The National Collegiate Cyber Defense Competition brings student experts together to the city to train and learn more about a growing industry.", "Inc. predicts the demand for cybersecurity will benefit San Antonio as globalspending is expected to hit $1 trillion within the next five years.", "The magazine's spotlight on San Antonio comes a few months after the Alamo City was named the best city for software developers in the entire country, based on job availability, average income and cost of living." ]

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[ "Local and metastatic growth and in vivo differentiation of human myeloid leukemia cells transplanted in nude mice.", "\nCells from the established human myeloid cell lines KG-1, KG-1a, and HL-60, transplanted subcutaneously (sc) into nude mice, developed discrete tumors (myelosarcomas). ", "These myelosarcomas had a host's age-associated pattern of growth identical to that of experimental tumors produced by sc transplantation of cells derived from malignant solid neoplasias. ", "Thus, leukemia cells yielded either localized myelosarcomas at the site of inoculation or a disseminated neoplastic growth after inoculation in adult (more than 4 weeks old) or newborn (1-3 days old) nude mice, respectively. ", "Human myeloid leukemia cells proliferating in the nude mice preserved the human karyotype and a surface antigenic determinant and did not influence the hematopoietic tissues of the host. ", "The KG-1 and HL-60 cell lines consistently attained varying degrees of differentiation along the myeloid series in vitro, and these features were maintained during proliferation in the mice. ", "Furthermore, cells of the variant subline KG-1a, which had a blastic morphology, developed signs of differentiation that were not seen in culture. ", "The presence of readily identifiable markers, such as cytoplasmic granules containing myeloperoxidase, in the cell lines tested makes these models particularly useful for studying the influence of a biological environment on cell differentiation and its influence on tumor growth. ", "These experimental systems are also suitable for investigating the mechanism(s) of metastases and for in vivo experimental therapeutic trials." ]

{ "pile_set_name": "PubMed Abstracts" }

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[ "The Buffalo Sabres are in the midst of one of the most important offseasons in the franchise’s 45-year history. ", "In addition to securing a top-tier player in the 2015 NHL Entry Draft — a player likely to be named Jack Eichel –, Buffalo has to find itself a new head coach to lead the team into the future. ", "General manager Tim Murray also has to continue to add talent to his roster to get the Sabres back to the playoffs for the first time since 2011.", "\n\nThe four-year playoff drought is the longest in Buffalo’s near half-century history.", "\n\nFrom the acquisition of former Winnipeg Jet goal scorer Evander Kane to the emergence of forward Zemgus Girgensons and defenseman Rasmus Ristolainen, the Sabres rebuild from two years of tanking is on a nice path upwards. ", "With the likely addition of Eichel, as well as last year’s second-overall selection Sam Reinhart, Buffalo is in good shape towards contending in the future. ", "But what players can the Sabres add this offseason to continue towards a successful rebuild?", "\n\nMichael Frolik — Right Wing\n\nFrom my posts on my personal twitter account to articles I’ve written in the past, those who follow me know exactly how I feel about Frolik and the idea of the Sabres signing him this summer. ", "He’s a player that can come aboard, join a line with Matt Moulson and, likely, Sam Reinhart and put up some pretty solid second-line numbers. ", "He’s had back-to-back 42 point seasons, and was a big reason why the Winnipeg Jets made it to the playoffs for the first time since returning to the NHL in 2011.", "\n\nHe’s scored over 40 points four times during his career, and there’s no reason to believe he couldn’t do the same, if not more, with more ice time that he would likely be given with the Sabres. ", "In terms of free agent forwards, I’d put Frolik at the top of the target list.", "\n\nAntti Niemi — Goalie\n\nThe Sabres need a goaltender; it’s priority position number one for Murray during the offseason. ", "Right now, Buffalo has Chad Johnson as the only goaltender under contract with more that 10 games of NHL experience. ", "Matt Hackett is a restricted free agent, and Anders Lindback is an unrestricted free agent.", "\n\nIt’s not the strongest class for goaltending, but Niemi represents the best of the available crop. ", "He’s put up solid numbers during his five years in San Jose, and has playoff experience to a team that really doesn’t have any. ", "If not him, the next best free agent target would be former Sabres goaltender Michal Neuvirth. ", "That being said, if Niemi isn’t the guy, it’s more likely that Murray trades for a goaltender rather than bringing back an old one.", "\n\nJustin Williams — Right Wing\n\nYou want to talk playoff experience, there is no one in this group of players with more than Justin Williams. ", "Even at 33 years old, the former Conn Smythe winner will certainly be a coveted player by many teams, including some potential contenders. ", "He’s missed just one game over the last four seasons, and has scored 70 goals during that span. ", "Though teams like Pittsburgh and Columbus could make a strong push for his services, the Sabres’ ability to offer Williams a larger role on the team could be a major selling point towards bringing him in.", "\n\nPaul Martin — Defense\n\nYes, the Sabres have a plethora of good — and great — defensemen coming up through the organization. ", "However, adding a player of Martin’s status would be a nice stop gap while giving the younger guys a bit more time to get ready for the NHL. ", "It’s expected that Mark Pysyk will finally join Ristolainen, Nikita Zadorov, Zach Bogosian and Josh Gorges with the Sabres full time this year, but someone has to take the place of Mike Weber. ", "Martin could be the guy.", "\n\nJoel Ward — Forward\n\nThis last target isn’t really high on the priority level, but he could be a nice addition should the Sabres not be able to add one of Frolik or Williams. ", "Ward is older than some people actually think (34) due to not becoming a full-time NHL player until age 28, but he could still produce when given the opportunity. ", "He was given his chance to show what he could do in Washington, and he put up a career best in goals, assists and points for a season during 2013-14 with 24 goals, 25 assists and 49 points. ", "If he can come in produce even close to that level, he would a very good second-line winger for Buffalo. ", "He would also likely command the lowest salary of the three forwards on this list." ]

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[ "The present invention relates to resistors adapted for use in electrical circuits and formed of aluminum-titanium alloys.", "\nHeavy duty power resistors are commonly employed in electrical circuits to control electrical current flow by converting electrical energy to heat, which may then be dissipated into the surrounding environment. ", "Normally, resistors rated at 300 watts and above are considered power resistors. ", "Typically, power resistors have been made from nickel-chromium alloys (NiChromes), copper-nickel alloys (Cuxe2x80x94Ni) or stainless steel alloys, with FeCrAl, 304 and 430 being the most common stainless steel types. ", "Stainless steel is often modified with additional metals to improve its electrical characteristics, for example, resistivity and changes in resistivity levels over an operating temperature range. ", "While all of these materials may be used in high temperature applications, i.e. up to about 1000xc2x0 C., they all have one or more shortcomings which compromise their use.", "\nFor example, the nickel-chromium alloys commonly referred to NiChrome materials are expensive and heavy, both of which factors limit their use in a wide range of applications. ", "On the other hand, the copper-nickel alloys are expensive and exhibit relatively low working temperatures and melting points. ", "Additionally, the copper-nickel alloys are disadvantageous in that they are not readily available in sheet form. ", "The stainless steel alloys also exhibit a relatively low resistivity and typically the resistivities of these alloys vary substantially over a temperature range, thereby rendering the alloys unsuitable for applications requiring precise resistivity requirements. ", "Additionally, the type 430 stainless steel which is commonly employed is slightly magnetic and therefore unsuitable for low inductance applications. ", "Various modified forms of stainless steel are also slightly magnetic and therefore unsuitable for low inductance applications. ", "These modified stainless steel alloys are also typically more expensive and therefore not attractive for widespread use.", "\nAccordingly, there is a continuing need for new resistors which would be suitable for widespread use, and particularly in heavy duty environments.", "\nAccordingly, it is an object of the present invention to provide new resistors which may be advantageously employed in various applications owing to a desirable combination of properties. ", "It is a further object of the invention to provide non-magnetic and lightweight resistors. ", "It is a related object to provide such resistors which can be rated at 10 watts and above and which may be employed in heavy duty power environments. ", "It is another object to provide resistors which may be employed in high temperature applications and in a temperature range of xe2x88x9240 to 1200xc2x0 C.\nThese and additional objects are satisfied by the present invention which is directed to resistors adapted for use in electrical circuits. ", "The resistors are formed of an alloy comprising from about 50 to 95 mol percent aluminum, from about 5 to about 50 mol percent titanium and up to about 15 mol percent of at least one additional metal or boron or a combination thereof. ", "The resistors according to the present invention are strong, lightweight and non-magnetic. ", "Additionally, the resistors according to the present invention exhibit nearly constant resistivity over a wide operating temperature range. ", "The alloys from which the resistors are formed exhibit a good combination of ductility, material density and melting point to allow efficient manufacture of the resistors.", "\nThese and additional objects and advantages provided by the present invention will be more fully understood in view of the following detailed description." ]

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[ "1. ", "Introduction {#sec1-ijms-21-00491}\n===============\n\n*Allium* vegetables such as garlic (*Allium sativum* L.) are rich in allyl sulfides that have been shown to prevent human chronic diseases, including cancer \\[[@B1-ijms-21-00491]\\]. ", "L-alliin (S-allyl-[l]{.smallcaps}-cysteine sulfoxide) is the major allyl sulfide component in garlic, which is converted into 2-propensulfenic acid by the endogenous enzyme alliinase, thereby producing the unstable thiosulfinate compound allicin (*S*-allyl-2-prop-2-ene-thiosulfinate). ", "Allicin readily breaks down to form organosulfur compounds (OSCs), including allyl sulfides such as diallyl trisulfide (DATS), diallyl disulfide (DADS), and diallyl sulfide (DAS), as well as ajoene and vinyl dithiins \\[[@B2-ijms-21-00491]\\]. ", "OSCs can modulate the immune system and attenuate inflammation, and these effects contribute to cancer prevention \\[[@B3-ijms-21-00491]\\]. ", "DATS, DADS, and DAS are known to prevent the well-characterized chemical-induced skin carcinogenesis \\[[@B4-ijms-21-00491],[@B5-ijms-21-00491],[@B6-ijms-21-00491]\\]. ", "Moreover, the OSCs have anticancer activity in a variety of cancer cell types, including skin cancer such as melanoma and basal cell carcinoma \\[[@B7-ijms-21-00491]\\]. ", "Of these, DATS is the most widely studied. ", "This compound induces apoptosis in a range of cancer cells, including malignant melanoma, osteosarcoma, and leukemia \\[[@B8-ijms-21-00491],[@B9-ijms-21-00491],[@B10-ijms-21-00491],[@B11-ijms-21-00491]\\]. ", "DATS also potentiates apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in human prostate cancer and melanoma cells \\[[@B12-ijms-21-00491],[@B13-ijms-21-00491],[@B14-ijms-21-00491]\\]. ", "The induction and amplification of apoptosis are associated with activation of caspases, upregulation of multiple pro-apoptotic molecules, including death receptor (DR)4/DR5, Bax, and Bak as well as downregulation of anti-apoptotic molecules such as Bcl-2 and Bcl-xL \\[[@B10-ijms-21-00491]\\]. ", "Fully activation of TRAIL-induced apoptosis in melanoma cells requires not only the intrinsic mitochondrial pathway but also the endoplasmic reticulum (ER) stress pathway \\[[@B14-ijms-21-00491]\\]. ", "The observations are consistent with the clinical findings that TRAIL can activate robust intrinsic death signaling but are ineffective in drug-resistant cancers, including malignant melanoma \\[[@B15-ijms-21-00491],[@B16-ijms-21-00491]\\]. ", "Strikingly, DADS and DATS exhibit the anticancer and TRAIL-sensitizing effects in a tumor-selective manner \\[[@B8-ijms-21-00491],[@B14-ijms-21-00491]\\], and they commonly induce the intracellular generation of reactive oxygen species (ROS) \\[[@B8-ijms-21-00491],[@B9-ijms-21-00491]\\]. ", "These observations support the view that oxidative stress plays a vital role in the cancer cell-selective killing and TRAIL sensitization \\[[@B17-ijms-21-00491],[@B18-ijms-21-00491],[@B19-ijms-21-00491]\\].", "\n\nCa^2+^ is a highly versatile intracellular second messenger that regulates numerous complicated cellular processes, including cell activation, proliferation, and death. ", "The increase in the cytosolic Ca^2+^ concentration (\\[Ca^2+^\\]~cyt~) results in the activation of Ca^2+^/calpain, an intracellular Ca^2+^-dependent cysteine protease, thereby resulting in the processing of the mitochondrial localized pro-apoptotic molecule, apoptosis-inducing factor, and caspase-independent apoptosis. ", "The activation of Ca^2+^/calpain occurs during the apoptosis of glioblastoma and neuroblastoma induced by DAS, DADS, and DATS \\[[@B8-ijms-21-00491],[@B9-ijms-21-00491]\\]. ", "Meanwhile, an excess rise in the mitochondrial Ca^2+^ concentration (\\[Ca^2+^\\]~mit~) leads to increased permeability of the mitochondrial membrane, mitochondrial dysfunction, and release of pro-apoptotic molecules such as cytochrome c and Apaf-1, resulting in caspase-dependent apoptosis. ", "Store-operated Ca^2+^ entry (SOCE) is the principal route by which external Ca^2+^ enters in non-excitable cells. ", "Ca^2+^ depletion in the endoplasmic reticulum (ER) triggers SOCE. ", "The depletion induces the translocation of stromal interaction molecules 1 (STIM1) to ER/plasma membrane (PM) junctional regions, where the molecule activates the PM channel ORAI1, leading to Ca^2+^ entry from the extracellular milieu. ", "Recently, Gualdani and colleagues reported that SOCE mediates cisplatin-induced Ca^2+^~mit~ overload, ROS production, and apoptosis \\[[@B20-ijms-21-00491]\\].", "\n\nMelittin, a 26-amino acid amphipathic peptide from bee venom, exhibits the anticancer effect against a variety of tumor cells, including melanoma, leukemia, osteosarcoma, lung, and bladder cancer cells. ", "Melittin induces apoptosis in these tumor cells in a caspase-dependent manner \\[[@B21-ijms-21-00491],[@B22-ijms-21-00491],[@B23-ijms-21-00491]\\]. ", "Moreover, earlier studies have suggested that melittin induces apoptosis through Ca^2+^ entry \\[[@B23-ijms-21-00491],[@B24-ijms-21-00491]\\]. ", "Notably, melittin is also known to activate SOCE via the activation of Ca^2+^-independent phospholipase A~2~ (iPLA~2~) \\[[@B25-ijms-21-00491],[@B26-ijms-21-00491]\\]. ", "Collectively, by analogy with cisplatin, it is plausible to speculate that melittin exhibits the anticancer effect through Ca^2+^ overload via SOCE. ", "While DATS preferentially acts on tumor cells \\[[@B8-ijms-21-00491],[@B14-ijms-21-00491]\\], melittin acts non-specifically, as cisplatin does. ", "Therefore, the identification of the Ca^2+^ transport pathways preferentially activated by DATS may help to define the Ca^2+^ pathways targeted in future tumor-selective therapy.", "\n\nIn the present study, we explored the ability of DATS to cause Ca^2+^ dysregulation and its possible role in the anti-melanoma effect using melittin as a reference. ", "We found that DATS explicitly induces massive Ca^2+^ dysregulation and apoptosis through a voltage-gated Ca^2+^ entry (VGCE) pathway that is distinct from SOCE.", "\n\n2. ", "Results {#sec2-ijms-21-00491}\n==========\n\n2.1. ", "DATS Reduces Cell Viability and Increases Apoptosis in a Ca^2+^- and Caspase-Dependent Manner {#sec2dot1-ijms-21-00491}\n--------------------------------------------------------------------------------------------------\n\nWe treated human malignant melanoma cell lines A375 and A2058 with varying concentrations of DATS for 72 h and measured cell viability by using the WST-8 assay. ", "Based on our previous study, we determined the range of the concentrations examined \\[[@B14-ijms-21-00491]\\]. ", "DATS (≥100 μM) decreased the viability of A375 cells in a dose-dependent manner ([Figure 1](#ijms-21-00491-f001){ref-type=\"fig\"}A). ", "DATS also reduced the viability of A2058 in a dose-dependent manner ([Figure 1](#ijms-21-00491-f001){ref-type=\"fig\"}B). ", "However, cellular sensitivity to DATS varied considerably in different experiments. ", "As a result, the IC~50~ values for A375 and A2058 cells were 159 ± 32 and 228 ± 60 μM, respectively (the mean ± SD, *n* = 3--6). ", "Flow cytometric analyses using annexin V and PI staining revealed that 72-h-treatment with DATS (100 μM) alone resulted in a massive increase in apoptotic (annexin V-positive) cells. ", "TRAIL markedly augmented the effect while the pan-caspase inhibitor Z-VAD-FMK entirely blocked it ([Figure 1](#ijms-21-00491-f001){ref-type=\"fig\"}C,D). ", "We found that Ca^2+^ was a critical regulator of drug sensitivity. ", "Treatment with the extracellular Ca^2+^ chelator EGTA (≤0.5 mM) or the intracellular Ca^2+^ chelator BAPTA had minimal effect on cell viability. ", "However, these chelators significantly reduced the anticancer effect of DATS in A375 and A2058 cells ([Figure 1](#ijms-21-00491-f001){ref-type=\"fig\"}E,F).", "\n\n2.2. ", "Melittin Exhibits Anti-Melanoma Effect in a Ca^2+^-Dependent Manner {#sec2dot2-ijms-21-00491}\n------------------------------------------------------------------------\n\nMelittin is known to be a potent inducer of apoptosis in melanoma cells. ", "Consistent with this view, treatment with the compound (≥2.5 μg/mL) for 72 h resulted in a robust increase in apoptotic (annexin V-positive) cells in A375 cells ([Figure 2](#ijms-21-00491-f002){ref-type=\"fig\"}A). ", "Meanwhile, the treatment minimally increased necrotic (annexin V-negative) cells. ", "The extracellular Ca^2+^ removal by EGTA (0.5 mM) augmented the effect of the subtoxic dose (1 μg/mL) of melittin. ", "On the other hand, it mitigated the increase in apoptosis while enhancing the increase in necrosis caused by the toxic concentration (5 μg/mL) of melittin ([Figure 2](#ijms-21-00491-f002){ref-type=\"fig\"}B).", "\n\n2.3. ", "DATS Increases \\[Ca^2+^\\]~mit~ without Increasing \\[Ca^2+^\\]~cyt~ {#sec2dot3-ijms-21-00491}\n----------------------------------------------------------------------\n\nNext, we determined whether DATS affected the intracellular Ca^2+^ level. ", "First, we tested the effect on \\[Ca^2+^\\]~cyt~. We used the Ca^2+^-ATPase inhibitor, thapsigargin (Tg), as a positive control, because it depletes the ER Ca^2+^ stores, thereby stimulating SOCE. ", "Tg substantially increased \\[Ca^2+^\\]~cyt~, while DATS at the concentration of up to 200 μM minimally increased it ([Figure 3](#ijms-21-00491-f003){ref-type=\"fig\"}A,B). ", "On the other hand, DATS increased \\[Ca^2+^\\]~mit~ in a dose-dependent manner ([Figure 3](#ijms-21-00491-f003){ref-type=\"fig\"}C,D). ", "This increase occurred immediately, developed over time, and the effect of DATS (200 μM) was even higher than that induced by the positive control A23187 (5 μM).", "\n\n2.4. ", "Melittin Increases Both \\[Ca^2+^\\]~cyt~ and \\[Ca^2+^\\]~mit~ in an Extracellular Ca^2+^-Dependent Manner {#sec2dot4-ijms-21-00491}\n------------------------------------------------------------------------------------------------------------\n\nThe site-specific Ca^2+^ measurements showed that melittin (1.3--5 μg/mL) increased both \\[Ca^2+^\\]~cyt~ and \\[Ca^2+^\\]~mit~ in a dose-dependent manner ([Figure 4](#ijms-21-00491-f004){ref-type=\"fig\"}A,C). ", "Removal of the extracellular Ca^2+^ entirely abolished the increase in \\[Ca^2+^\\]~cyt~ but partially reduced the increase in \\[Ca^2+^\\]~mit~, indicating that these effects depend on the extracellular Ca^2+^ influx to different degrees ([Figure 4](#ijms-21-00491-f004){ref-type=\"fig\"}B,D).", "\n\n2.5. ", "Acidification Has a Reciprocal Effect on Ca^2+^~mit~ Overload Caused by DATS and Melittin {#sec2dot5-ijms-21-00491}\n----------------------------------------------------------------------------------------------\n\nThe microenvironments, including the extracellular pH conditions, contribute to the various aspects of malignant phenotypes of cancer cells \\[[@B27-ijms-21-00491],[@B28-ijms-21-00491]\\]. ", "Therefore, we examined the effect of altered extracellular pH on Ca^2+^~mit~ overload. ", "Cells were suspended in HBSS with different pH 6.8 and 7.4, treated with DATS, and measured for the intracellular Ca^2+^ levels. ", "Notably, after DATS treatment, a small but significant increase in \\[Ca^2+^\\]~cyt~ was seen at pH 6.8, but not pH 7.4 ([Figure 5](#ijms-21-00491-f005){ref-type=\"fig\"}A,B). ", "Similarly, a greater extent of Ca^2+^~mit~ overload occurred at pH 6.8 compared to pH 7.4, and this impact was more pronounced for the higher concentration of the compound ([Figure 5](#ijms-21-00491-f005){ref-type=\"fig\"}C,D). ", "The effect was apparent for higher concentrations of DATS. ", "Next, we analyzed the effect of the extracellular Ca^2+^ removal on Ca^2+^~mit~ overload seen under different pH. Cells were suspended in HBSS (pH 7.4 or 6.8) containing CaCl~2~ or EGTA and treated with DATS. ", "At pH 7.4, the Ca^2+^ removal almost entirely abolished the increase in \\[Ca^2+^\\]~mit~, while at pH 6.8, it reduced the increase only a modestly ([Figure 5](#ijms-21-00491-f005){ref-type=\"fig\"}E,F). ", "In contrast to DATS, acidification significantly mitigated Ca^2+^~mit~ overload caused by melittin. ", "A much smaller increase in \\[Ca^2+^\\]~mit~ occurred at pH 6.8 compared to at pH 7.4 in response to a range of concentrations of melittin (1.3--5 μg/mL). [", "Figure 5](#ijms-21-00491-f005){ref-type=\"fig\"}G shows the data for melittin (2.5 μg/mL). ", "Notably, we observed a comparable level of \\[Ca^2+^\\]~mit~ increase in the cells placed in HBSS (pH 7.4) and Ca^2+^-containing buffer. ", "The \\[Ca^2+^\\]~mit~ rise seen in HBSS (pH 6.8) and Ca^2+^-free buffer (pH 7.4) after melittin treatment were also comparable ([Figure 5](#ijms-21-00491-f005){ref-type=\"fig\"}G,H).", "\n\n2.6. ", "Acidification Mitigates SOCE {#sec2dot6-ijms-21-00491}\n---------------------------------\n\nThe data presented so far indicated that acidification reduced Ca^2+^~mitt~ overload caused by melittin. ", "Since melittin is known as a potent activator of SOCE, we determined the effect of acidification on SOCE. ", "We activated SOCE by the Ca^2+^ depletion-Ca^2+^ re-addition technique and examined the impact of acidification on the Ca^2+^ entry. ", "At neutral extracellular pH, the addition of Tg to cells in a Ca^2+^-free medium resulted in a small and transient increase in \\[Ca^2+^\\]~cyt~, indicating the Ca^2+^ release from the ER. ", "The re-addition of Ca^2+^ to Tg-treated cells led to a higher and persistent increase in \\[Ca^2+^\\]~cyt~, and the inositol-1,4,5-triphosphate receptor (IP~3~R)/SOCE antagonists 2-aminoethoxydiphenyl borate (2-APB) entirely blocked this Ca^2+^ response, verifying the onset of SOCE ([Figure 6](#ijms-21-00491-f006){ref-type=\"fig\"}A). ", "Even without any Ca^2+^ depletion in the ER (i.e., in the absence of Tg), a smaller but significant \\[Ca^2+^\\]~cyt~ rise was seen following Ca^2+^ re-addition and was minimally affected by 2-APB ([Figure 6](#ijms-21-00491-f006){ref-type=\"fig\"}A, Right), indicating the onset of a store-independent Ca^2+^ influx (non-SOCE). ", "Meanwhile, at pH 6.8, no substantial SOCE was observed, while the non-SOCE was increased by the acidification ([Figure 6](#ijms-21-00491-f006){ref-type=\"fig\"}B).", "\n\n2.7. ", "Ca^2+^ Channel Blockers Inhibit Ca^2+^~mit~ Overload and Cell Death Caused by DATS {#sec2dot7-ijms-21-00491}\n---------------------------------------------------------------------------------------\n\nTo gain insight into the transport pathway(s) involved Ca^2+^~mitt~ overload caused by DATS, we look for Ca^2+^ channel antagonists affecting the \\[Ca^2+^\\]~mit~ rise. ", "Eventually, we found that the Ca^2+^ channel blockers nifedipine, verapamil, and diltiazem significantly inhibited the effect of DATS. ", "The inhibitory effect was seen more pronouncedly at pH 6.8 than at pH 7.4 ([Figure 7](#ijms-21-00491-f007){ref-type=\"fig\"}A). ", "Under the acidic conditions, nifedipine exhibited the most potent effect (70% reduction), while the other compounds inhibited it around 50% ([Figure 7](#ijms-21-00491-f007){ref-type=\"fig\"}A). ", "2-APB had an inhibitory effect comparable to verapamil and diltiazem. ", "Meanwhile, at the neutral pH conditions, these inhibitors mitigated the \\[Ca^2+^\\]~mit~ rise only modestly (20%; [Figure 7](#ijms-21-00491-f007){ref-type=\"fig\"}A). ", "Next, we examined the effect of Ca^2+^ blockers on the anti-melanoma effect in highly growing cells. ", "Nifedipine, verapamil, and diltiazem significantly inhibited the anti-melanoma effect of DATS (200 μM; [Figure 7](#ijms-21-00491-f007){ref-type=\"fig\"}B).", "\n\n3. ", "Discussion {#sec3-ijms-21-00491}\n=============\n\nIn this study, we elucidated the possible role of Ca^2+^ dysregulation in the anti-melanoma effect of DATS with a particular interest in the Ca^2+^ pathways involved. ", "We found that DATS induced caspase-dependent apoptosis in melanoma cells, although relatively high concentrations of DATS are required ([Figure 1](#ijms-21-00491-f001){ref-type=\"fig\"}). ", "These results are consistent with previous studies, including our own, which demonstrate that DATS induces apoptosis in a variety of cancer cell types \\[[@B8-ijms-21-00491],[@B9-ijms-21-00491],[@B10-ijms-21-00491],[@B11-ijms-21-00491],[@B14-ijms-21-00491]\\]. ", "We noticed that cell confluency considerably affected the sensitivity to the compound. ", "Cells growing near confluent tended to be more susceptible than those growing at low confluency. ", "In agreement with the previous reports by others \\[[@B23-ijms-21-00491],[@B24-ijms-21-00491]\\], melittin also primarily induced apoptosis in the cells ([Figure 2](#ijms-21-00491-f002){ref-type=\"fig\"}A). ", "Moreover, we found that extracellular Ca^2+^ entry was necessary for apoptosis caused by DATS and melittin ([Figure 1](#ijms-21-00491-f001){ref-type=\"fig\"}E,F and [Figure 2](#ijms-21-00491-f002){ref-type=\"fig\"}B). ", "The Ca^2+^~mit~ uptake is generally due to so-called reservoir function of the organelles: the mitochondria take up Ca^2+^ in response to increased \\[Ca^2+^\\]~cyt~ thereby preventing excessive \\[Ca^2+^\\]~cyt~ rise while they release Ca^2+^ in response to decreased \\[Ca^2+^\\]~cyt~ thereby counteracting excessive \\[Ca^2+^\\]~cyt~ depletion. ", "Notably, we found that DATS and melittin had different impacts on \\[Ca^2+^\\]~cyt~ and \\[Ca^2+^\\]~mit~. DATS substantially increased \\[Ca^2+^\\]~mit~, but not \\[Ca^2+^\\]~cyt~ ([Figure 3](#ijms-21-00491-f003){ref-type=\"fig\"}), suggesting that the Ca^2+^~mit~ overload is a result of the activation of a specific Ca^2+^ transport pathway rather than a simple Ca^2+^~mit~ uptake. ", "Meanwhile, melittin markedly increased \\[Ca^2+^\\]~cyt~ and \\[Ca^2+^\\]~mit~ ([Figure 4](#ijms-21-00491-f004){ref-type=\"fig\"}). ", "However, extracellular Ca^2+^ removal entirely abolished the former, while attenuated the latter partially ([Figure 4](#ijms-21-00491-f004){ref-type=\"fig\"}B,D). ", "These results suggest that the \\[Ca^2+^\\]~cyt~ rise primarily results from extracellular Ca^2+^ entry while the \\[Ca^2+^\\]~mit~ increase is due to Ca^2+^ transport of both extracellular and intracellular Ca^2+^. Dysregulation of \\[Ca^2+^\\]~mit~ is a master cause of cell death. ", "A fine-tuned increase in \\[Ca^2+^\\]~mit~ supports energy metabolism, cell activation, and cell survival, while Ca^2+^~mitt~ overload can damage mitochondrial integrity, thereby inducing mitochondrial permeability transition pore (MPTP) opening and the resulting release of apoptogenic proteins \\[[@B28-ijms-21-00491],[@B29-ijms-21-00491],[@B30-ijms-21-00491]\\]. ", "Collectively, we conclude that different Ca^2+^ transport pathways participate in the Ca^2+^~mitt~ overload and apoptosis caused by DATS and melittin ([Figure 8](#ijms-21-00491-f008){ref-type=\"fig\"}).", "\n\nStrikingly, DATS seems to evoke Ca^2+^~mit~ overload through different Ca^2+^ pathways, depending on the extracellular pH. At pH 7.4, DATS exhibits the effect mainly via the extracellular Ca^2+^ entry ([Figure 5](#ijms-21-00491-f005){ref-type=\"fig\"}E), while at pH 6.8, DATS seemed to use both extracellular and intracellular Ca^2^ entry pathways ([Figure 5](#ijms-21-00491-f005){ref-type=\"fig\"}F). ", "Ca^2+^ released from the IP~3~R can easily reach the mitochondrial matrix via the IP~3~R--voltage-dependent anion channel (VDAC1)--mitochondrial Ca^2+^ uniporter (MCU) pathway \\[[@B31-ijms-21-00491],[@B32-ijms-21-00491]\\]. ", "The ER is the primary mechanism for regulating \\[Ca^2+^\\]~mit~ and tethered to mitochondria via a mitochondria-associated membrane; the physical association allows rapid Ca^2+^ transport through specified microdomains \\[[@B33-ijms-21-00491],[@B34-ijms-21-00491]\\]. ", "The IP~3~R physically links to VDAC1 on the outer mitochondrial membrane and transports Ca^2+^ into the mitochondrial matrix \\[[@B35-ijms-21-00491],[@B36-ijms-21-00491]\\]. ", "We previously showed that Ca^2+^~mit~ remodeling in osteosarcoma cells was under the control of Ca^2+^ uptake through the MCU \\[[@B37-ijms-21-00491]\\]. ", "\\[Ca^2+^\\]~mit~ is also regulated by Ca^2+^~mit~ efflux through several different pathways, including the mitochondrial Na^+^/Ca^2+^ exchanger (NCLX), Ca^2+^/H^+^ antiporter, and MPTP \\[[@B38-ijms-21-00491],[@B39-ijms-21-00491],[@B40-ijms-21-00491],[@B41-ijms-21-00491],[@B42-ijms-21-00491]\\]. ", "Notably, our previous study showed that the NCLX inhibitor CGP-37157 causes Ca^2+^~mit~ overload and apoptosis in osteosarcoma cells \\[[@B43-ijms-21-00491]\\]. ", "Therefore, increased Ca^2+^ transport via the IP~3~R--VDAC1--MCU pathway and decreased Ca^2+^~mitt~ extrusion could contribute to the Ca^2+^~mitt~ overload caused by DATS.", "\n\nSOCE is the most ubiquitous pathway for Ca^2+^ transport from the extracellular space and activated following the depletion of Ca^2+^ stores from the ER. ", "Recently, SOCE has emerged as critical machinery for Ca^2+^ influx, contributing to various malignant phenotypes in cancer cells \\[[@B44-ijms-21-00491],[@B45-ijms-21-00491],[@B46-ijms-21-00491]\\]. ", "SOCE is also shown to be responsible for cisplatin-induced apoptosis \\[[@B20-ijms-21-00491]\\]. ", "It is noteworthy that melittin can activate iPLA~2~ to produce arachidonic acid and lysophospholipids and activates SOCE in vascular smooth muscle cells \\[[@B25-ijms-21-00491],[@B26-ijms-21-00491]\\]. ", "Given that melittin activates SOCE in melanoma cells, too, this action can expect to cause massive Ca^2+^ dysregulation, as cisplatin does. ", "A notable property of SOCE is its sensitivity to acidification; SOCE is suppressed by decreasing extracellular pH in several cell types \\[[@B44-ijms-21-00491],[@B45-ijms-21-00491]\\]. ", "In agreement with these reports, acidification entirely abolished SOCE in melanoma cells ([Figure 6](#ijms-21-00491-f006){ref-type=\"fig\"}). ", "Notably, the Ca^2+^~mit~ overload induced by melittin was mainly dependent upon extracellular Ca^2+^ entry and was suppressed by acidification ([Figure 5](#ijms-21-00491-f005){ref-type=\"fig\"}). ", "Extracellular Ca^2+^ removal and acidification caused a similar degree of reduction, suggesting that extracellular Ca^2+^ entry is specifically sensitive to acidification, as observed with SOCE. ", "Together, our findings strongly suggest that melittin evokes Ca^2+^~mit~ overload via SOCE activation. ", "This view is consistent with other reports demonstrating the requirement of SOCE for active Ca^2+^~mit~ uptake \\[[@B47-ijms-21-00491],[@B48-ijms-21-00491]\\]. ", "However, further studies, including the examination of the involvement of SOCE components such as STIM1 and ORAI1, are necessary for validation.", "\n\nAnother important finding in the present study is that Ca^2+^ channel blockers significantly mitigate Ca^2+^~mit~ overload and the anti-melanoma effect of DATS at acidic pH ([Figure 7](#ijms-21-00491-f007){ref-type=\"fig\"}). ", "These findings suggest a role of VGCE in Ca^2+^~mit~ overload and apoptosis. ", "Notably, acidification increased Ca^2+^~mit~ overload caused by DATS ([Figure 5](#ijms-21-00491-f005){ref-type=\"fig\"}), and sensitivity to Ca^2+^ channel blockers was higher at acidic pH than at neutral pH ([Figure 7](#ijms-21-00491-f007){ref-type=\"fig\"}A). ", "The latter finding seems to provide another evidence for the role of VGCE because Kato and colleagues have shown that an acidic pH (5.4--6.5) increases Ca^2+^ influx in mouse B16 melanoma cells through VGCE \\[[@B49-ijms-21-00491]\\]. ", "The activation of VGCE requires plasma membrane depolarization, which leads to the loss of the negative membrane potential, the driving force transporting extracellular Ca^2+^ into inner cells. ", "Accordingly, SOCE could be compromised under these conditions. ", "Thus, acidification activates VGCE while mitigates SOCE, thereby possibly playing a vital role in switching the Ca^2+^ entry pathway from SOCE to VGCE. ", "The microenvironments of tumor cells are acidic due to an H^+^ efflux caused by increased vacuolar ATPase activity, and this acidification causes multiple malignant phenotypes, including increased proliferation, drug resistance, and metastasis \\[[@B27-ijms-21-00491],[@B28-ijms-21-00491]\\]. ", "Therefore, our observations that high confluent cells are relatively high responders of DATS could be explained by the predominant role of VGCE under acidic extracellular microenvironments, because the extracellular pH drops in cells growing at high density.", "\n\nThere remain issues to be resolved in future studies. ", "In particular, the identification of voltage-gated Ca^2+^ channels (VGCCs) involved in Ca^2+^~mit~ overload caused by DATS is challenging. ", "Our preliminary experiments revealed that melanoma cells expressed Ca~v~1.2 and Ca~v~1.3 isoform of [l]{.smallcaps}-type VGCCs. ", "Notably, the previous studies demonstrate the occurrence of the coordinate control of STIM1-ORAI1 and Ca~v~1.2 \\[[@B50-ijms-21-00491],[@B51-ijms-21-00491]\\]. ", "Therefore, there is an intriguing possibility that Ca~v~1.2 plays a role in Ca^2+^~mit~ overload via a specific interaction with SOCE. ", "Further studies are necessary to clarify the molecular entity of VGCCs involved and to test this hypothesis.", "\n\nIn conclusion, the present study demonstrated that DATS and melittin exhibit the anti-melanoma effects by evoking pro-apoptotic Ca^2+^~mit~ overload through the different pathways, possibly VGCE and SOCE. ", "Our findings suggest the involvement of multiple Ca^2+^ entry pathways in Ca^2+^~mit~ remodeling in melanoma cells and switching between them, depending on apoptotic stimuli and pH microenvironments.", "\n\n4. ", "Materials and Methods {#sec4-ijms-21-00491}\n========================\n\n4.1. ", "Materials {#sec4dot1-ijms-21-00491}\n--------------\n\nAll chemicals were purchased from Sigma Aldrich (St. Louis, MO, USA) unless otherwise specified. ", "Alliin and allicin were obtained from Cayman Chemical Co. (Ann Arbor, MI, USA). ", "Soluble recombinant human TRAIL was obtained from Enzo Life Sciences (San Diego, CA, USA). ", "The pan-caspase inhibitor Z-VAD-FMK was purchased from Merck Millipore (Darmstadt, Germany). ", "All insoluble reagents were dissolved in dimethylsulfoxide (DMSO) and diluted with high glucose-containing Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) or Hank's balanced salt solution (HBSS; pH 7.4, Nissui Pharmaceutical Co., Ltd., Tokyo, Japan; final DMSO concentration, \\<0.1%) prior to use.", "\n\n4.2. ", "Cell Culture {#sec4dot2-ijms-21-00491}\n-----------------\n\nCells were cultured in 10% FBS/DMEM supplemented with 100 U/mL penicillin and 100 μg streptomycin (Pen-Strep, Thermo Fisher Scientific Japan, Tokyo, Japan) in a 95% air/5% CO~2~ humidified atmosphere unless otherwise specified. ", "The human melanoma cell lines A375 (cell number CRL-1619) and A2058 (cell number IFO 50276) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and the Japanese Collection of Research Bioresources (JCRB) Cell Bank of National Institutes of Biomedical Innovation, Health, and Nutrition (Osaka, Japan), respectively. ", "Cells were harvested by incubation with 0.25% trypsin-ethylenediaminetetraacetic acid (EGTA; Thermo Fisher Scientific, Waltham, MA, USA) for 5 min at 37 °C.", "\n\n4.3. ", "Cell Viability Assay {#sec4dot3-ijms-21-00491}\n-------------------------\n\nCell viability was measured by WST-8 assay using Cell Counting Reagent SF (Nacalai Tesque, Inc., Kyoto, Japan) as previously described \\[[@B43-ijms-21-00491]\\] with modifications. ", "This method is a colorimetric assay based on the formation of a water-soluble formazan product. ", "Briefly, cells were seeded at a density of 8 × 10^3^ cells/well in 96-well plates (Corning Incorporated, Corning, NY, USA) and cultured with agents to be tested for 72 h at 37 °C before the addition of 10 μL of cell counting reagent SF and further incubation for 2 h. The absorbances were measured at 450 nm using an ARVO MX microplate reader (PerkinElmer Japan Co., Ltd., Yokohama, Japan).", "\n\n4.4. ", "Cell Death Assay {#sec4dot4-ijms-21-00491}\n---------------------\n\nCell death was quantitatively assessed by double-staining with fluorescein isothiocyanate (FITC)-conjugated annexin V and propidium iodide (PI) as previously described \\[[@B37-ijms-21-00491]\\]. ", "Briefly, cells plated in 24-well plates (2 × 10^5^ cells/well) were treated with the agents to be tested for 20 h and stained with FITC-conjugated annexin V and PI using a commercially available kit (Annexin V FITC Apoptosis Detection Kit I: BD Biosciences, San Jose, CA, USA). ", "The stained cells (10,000 cells) were analyzed in FL-1 and FL-2 channel of a FACSCalibur flow cytometer (BD Biosciences) using the CellQuest software (BD Biosciences). ", "Four cellular subpopulations were evaluated: viable cells (annexin V-negative and PI-negative); early apoptotic cells (annexin V-positive and PI-negative); late apoptotic cells (annexin V-positive and PI-positive); and necrotic/membrane-damaged cells (annexin V-negative and PI-positive).", "\n\n4.5. ", "Intracellular Ca^2+^ Measurements {#sec4dot5-ijms-21-00491}\n--------------------------------------\n\nChanges in Ca^2+^~cyt~ and Ca^2+^~mit~ levels were measured using Fluo4-AM and rhod2-AM (Dojindo Kumamoto, Japan), respectively, as previously described \\[[@B37-ijms-21-00491]\\]. ", "For improvement of mitochondrial localization of rhod 2-AM \\[[@B52-ijms-21-00491]\\], it was reduced to the colorless, nonfluorescent dihydrorhod 2-AM by sodium borohydride, according to the manufacturer's protocol. ", "Cells were loaded with 4 μM each of Fluo 4-AM or dihydrorhod 2-AM for 40 min at 37 °C, washed with HBSS. ", "Then, the cells (1 × 10^6^/mL) were resuspended in HBSS in 96-well plates. ", "The cells were manually added with the agents to be tested. ", "Then, the cells were measured for fluorescence in a microplate reader (Fluoroskan Ascent, ThermoFisher Scientific) with excitation and emission at 485 and 538 nm (for Fluo 4-AM), respectively, and 542 and 592 nm (for rhod 2-AM), respectively. ", "For the analysis of Ca^2+^ release and SOCE, Fluo4-AM-loaded cells were suspended in a Ca^2+^-free medium (HBSS supplemented with 1 mM EGTA) and treated with 2 μM Tg for 10 min and then added to 2 mM CaCl~2~. For analysis of non-SOCE, cells in the Ca^2+^-free medium were treated with the medium for 10 min and then added to 2 mM CaCl~2~.\n\n4.6. ", "Statistical Analysis {#sec4dot6-ijms-21-00491}\n-------------------------\n\nData are presented as mean ± standard deviation (SD) or standard error (SE) and were analyzed by one-way analysis of variance followed by Tukey's post hoc test using add-in software for Excel 2016 for Windows (SSRI, Tokyo, Japan)*. ", "p* \\< 0.05 was considered statistically significant.", "\n\nWe thank the JCRB Cell Bank of National Institutes of Biomedical Innovation, Health, and Nutrition (Osaka, Japan) and the Riken BioResource Center (Tsukuba, Japan) for providing cell lines.", "\n\nConceptualization, Y.S.-K.; methodology, T.O. and Y.S.-K.; validation, C.N., M.S.-K. (Manami Suzuki-Karasaki) and Y.S.-K.; investigation, C.N., M.S.-K. (Manami Suzuki-Karasaki), M.S.-K. (Miki Suzuki-Karasaki) and Y.S.-K.; resources, T.O.; data curation, C.N.; writing---original draft preparation, C.N., M.S.-K. (Manami Suzuki-Karasaki) and Y.S.-K.; writing---review and editing, T.O. and Y.S.-K.; supervision, T.O. and Y.S.-K.; project administration, T.O. and Y.S.-K.; funding acquisition, T.O. and Y.S.-K. All authors have read and agreed to the published version of the manuscript.", "\n\nThis research was funded by JSPS KAKENHI, grant number 15K09792, and 18K09121.", "\n\nM.S.-K. (Manami Suzuki-Karasaki), M.S.-K. (Miki Suzuki-Karasaki), and Y.S.-K. are employees of the non-profit Institute Plasma ChemiBio Laboratory. ", "Other authors have no conflicts of interest. ", "The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.", "\n\n{#ijms-21-00491-f001}\n\n{ref-type=\"fig\"}. ", "Data represent the mean ± SD (*n* = 3). \\* *", "p* \\< 0.05; \\*\\*\\* *p* \\< 0.001.](ijms-21-00491-g002){#ijms-21-00491-f002}\n\n###### \n\nDATS induces Ca^2+^~mit~ overload with minimal Ca^2+^~cyt~ increase in melanoma cells. (**", "A**,**B**) DATS has minimal effect on \\[Ca^2+^\\]~cyt~. A375 cells were loaded with Fluo4-AM, added with DATS (50, 100, and 200 μM) or Tg (1 μM), and immediately measured for fluorescence for 3 min with excitation and emission at 485 and 538 nm, respectively, in a microplate fluorescence reader. ", "\\[Ca^2+^\\]~cyt~ was evaluated as described in Materials and Methods. (**", "C**,**D**) DATS induces Ca^2+^~mit~ overload in a dose-dependent manner. ", "The cells were loaded with dihydrorhod 2-AM, treated with DATS as described above**,** and measured for fluorescence with excitation and emission at 542 and 592 nm, respectively. ", "Data are shown as *F/F~0~*~,~ where *F* and *F~0~* represent the fluorescence of the sample and control (medium-treated cells), respectively. ", "Data shown in (**D**) represent the mean ± SE (*n* = 3). *\\*\\*\\* ", "p* \\< 0.001 vs. untreated control.", "\n\n![](", "ijms-21-00491-g003a)\n\n![](", "ijms-21-00491-g003b)\n\n{#ijms-21-00491-f004}\n\n###### \n\nAcidification has a reciprocal effect on Ca^2+^~mit~ overload caused by DATS and melittin. (**", "A**,**B**) A375 cells were suspended in HBSS (pH 7.4) (**A**) and 6.8 (**B**), treated with the indicated concentrations of DATS, and measured for fluorescence with excitation and emission at 485 and 538 nm, respectively in a microplate fluorescence reader. (**", "C**,**D**) Dihydrorhod2-AM-loaded A375 cells were suspended in HBSS (pH 7.4 or 6.8), treated with DATS (100 and 200 μM) and measured for fluorescence with excitation and emission at 542 and 592 nm, respectively, in a microplate fluorescence reader. ", "Data are shown as F/F~0~, where F and F~0~ represent the fluorescence of the sample and control, respectively (*n* = 3--5). ", "The data shown in (**D**) represent mean ± SD of the area under the curve (AUC) for 3 min (*n* = 3). (**", "E**,**F**) The cells were suspended in HBSS, pH 7.4 (**E**) or 6.8 (**F**) containing 1 mM CaCl~2~ (Ca+) or 1 mM EGTA (Ca−), treated with DATS (200 μM), and measured for the fluorescence as described above. ", "Data are shown as relative fluorescence unit (RFU). (**", "G**,**H**) The cells were suspended in HBSS (pH 7.4 or 6.8) (**G**) or HBSS (pH 7.4) containing 1 mM CaCl~2~ (Ca+) or 1 mM EGTA (Ca−) (**H**), treated with melittin (2.5 μg/mL) and measured for the fluorescence. \\* *", "p* \\< 0.05.", "\n\n![](", "ijms-21-00491-g005a)\n\n![](", "ijms-21-00491-g005b)\n\n###### \n\nAcidification mitigates store-operated Ca^2+^ entry (SOCE). ", "Fluo4-AM-loaded A375 cells were suspended in a Ca^2+^-free HBSS supplemented with 1 mM EGTA at pH 7.4 (**A**) or pH 6.8 (**B**), added with thapsigargin (Tg, 2 μM, red arrow) and incubated for 10 min to deplete intracellular Ca^2+^ stores. ", "Then, two mM Ca^2+^ was added to the cells (SOCE). ", "After the addition of Tg, the fluorescence was monitored with excitation and emission at 485 and 538 nm, respectively. ", "For validating SOCE, the cells in the Ca^2+^-free buffer were treated as described above in the presence of 2-APB (10 μM). ", "For non-SOCE measurement, the cells in the Ca^2+^-free buffer were treated with the medium and then added with 2 mM Ca^2+^ (non-SOCE). ", "Note that at pH 7.4, a higher increase in \\[Ca^2+^\\]~cyt~ was seen in the SOCE trace compared to the non-SOCE trace, and this effect was entirely abolished by 2-APB. ", "Meanwhile, no such effect was seen at pH 6.8.", "\n\n![](", "ijms-21-00491-g006a)\n\n![](", "ijms-21-00491-g006b)\n\n{#ijms-21-00491-f007}\n\n{#ijms-21-00491-f008}\n" ]

{ "pile_set_name": "PubMed Central" }

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[ "Detective Parker Bode with the Honolulu Police Department will answer some of your questions from our viewers on today’s Ask HPD:\n\n1. ", "Question: I hear vehicle alarms go off all the time, sometimes for minutes at a time. ", "can I call the police?", "\n\nAnswer: Yes, you can call 911 and an officer will be sent to your location. ", "there are many things that can activate vehicle alarms, and we understand that the constant noise can be annoying. ", "If an officer is called to investigate an excessive vehicle alarm and finds that the alarm is continually activated for more than five minutes, the registered owner can face a $112 citation. ", "If the vehicle alarm does not reset or stop, and continues to sound for more than 10 minutes, the vehicle can be removed from either public or private property.", "\n\n2. ", "Question, from Jeff: My neighbor’s dog barks at all hours of the day. ", "what can i do about this?", "\n\nAnswer: It’s against City and County of Honolulu law for a dog to bark constantly for 10 minutes, or unprovoked for 30 minutes on and off, to the disturbance of others. ", "You can call 911 and an officer will be sent to your location, or you can contact the Hawaiian humane society. ", "quiet neighborhoods are important to everyone. ", "Unresolved and persistent barking can lead to fines and court appearances for the owner.", "\n\n3. ", "Question from Dennis: What are the restrictions on a loud party?", "\n\nAnswer: If you feel the noise level of a party is unreasonable call HPD and report it. ", "officers will respond and determine if the noise level is reasonable given the time of day and location. ", "If the noise level is unreasonable, the officer will inform the party involved and the action needed for compliance. ", "a citation can be issued for non-compliance to a noise violation.", "\n\nIf you have a question you’d like to ask HPD, email it to askhpd@honolulu.gov or message us on social media." ]

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[ "Stuff happened, things changed.", "\n\nmcl2k2 pools and the em conversion. ", "The details are in the commits, but the short story is that due to hardware limitations, a number of tradeoffs need to be made between performance and memory usage. ", "The em chip can (mostly) only be programmed to write to 2k buffers. ", "However, ethernet payloads are not nicely aligned. ", "They’re two bytes off. ", "Leading to a costly choice. ", "Provide a 2k buffer, and then copy all the data after the fact, which is slow. ", "Or allocate a larger than 2k buffer, and provide em with a pointer that’s 2 bytes offset. ", "Previously, the next size up from 2k was 4k, which is quite wasteful. ", "The new 2k2 buffer size still wastes a bit of memory, but much less.", "\n\nOne less hint for malloc. ", "A forgotten holdover from another era. ", "Zap P too.", "\n\nFreeType 2.7 is prettier than ever.", "\n\nvmm for i386. ", "Improve security. ", "vmm is still running with a phenomenal set of privileges, but perhaps some cross-VM attacks may be limited. ", "On the other side of the world, hyperv support is getting better.", "\n\nFix an installer bug relating to user separation and local sets. ", "And one for the prefetch area. ", "The installer continues to get better and safer in mostly invisible ways. ", "Usually it’s only the bugs that get noticed.", "\n\nRemove setlocale. ", "setlocale was sprinkled all throughout the code base many years ago, even though it did nothing, in anticipation of a day when it would do something. ", "We’ve since decided that day will never come, and so many setlocale calls can go.", "\n\nThere’s a new build system. ", "It kind of works like before, but a lot of the details have changed to support less root. ", "Actually, it’d be accurate to say the whole build privilege system has been flipped. ", "Start as root, which drops down to the build user to do the heavy lifting, instead of starting as a user that can elevate to root at any time. ", "This no longer requires the build user to be pseudo-root; in fact, the goal is that the build user can’t elevate.", "\n\nUse x2APIC. ", "Some systems, notably VMs, require this and don’t provide the older interfaces.", "\n\nsyspatch is coming. ", "Lots of commits actually. ", "Despite the name, it’s more like a system update, since it replaces entire binaries. ", "Then again, replacing a few binaries in a system is like patching small parts of the whole. ", "A syspatch update will be smaller than an entire release.", "\n\nThe slab is back. ", "Per CPU caches existed in pool long ago, but were deleted while making the code simpler. ", "The pool code has been worked over quite thoroughly since then, and now it’s time to bring back this optimization. ", "Bonwick’s paper is a systems classic. ", "For reference, ancient commit introducing caches previously.", "\n\nOCSP client support in libtls. ", "And therefore nc as well.", "\n\nThe libcrypto manuals are getting a makeover. ", "The other pages were converted as well.", "\n\nlibX11 was updated. ", "Lots of other libraries, too. ", "All the newest hotnesses. ", "xterm 327.", "\n\nAdd some perl, I mean assembly, code for crypto curves. ", "Add X25519 curve support. ", "Better support for verifying cert chains.", "\n\nxenodem gets a feature: autologin.", "\n\nThe thread process deunification continues. ", "Threads started out as clumps of processes. ", "Slowly but surely, things are being pried apart into distinct concepts.", "\n\nPreparatory steps to making network stack lock safe. ", "The old spl model is recursive, which won’t work in the future. ", "More discipline is now required to know who is responsible for acquiring and releasing exclusion. ", "It seems like only yesterday I rewrote sblock to be an rwlock, and now sblock is back to using tsleep again.", "\n\nswitch and switchd are getting better. ", "openflow in switchctl." ]

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[ "Q:\n\nissue with selecting 1 or 0 in mysql db\n\nI am having trouble with the following SQL statement. ", "I have had this issue before but I can't remember how I fixed the problem. ", "I am guessing the issue with this is that MySQL sees 0 as null? ", "Note I didn't show the first part of the Select statement as it's irrelevant. ", "My SQL works. ", "It's showing rows with toffline that are = to 1 as well as 0...\nFROM table1 AS tb1\nLEFT JOIN table2 AS tb2 ON tb2.id = tb1.id2\nLEFT JOIN table3 AS tb3 ON tb3.id = tb1.id3\nAND tb1.toffline = 0\n\nI have also tried AND NOT tb1.toffline = 1 also WHERE tb1.toffline = 0 all with the same result...\n\nA:\n\nThis is where indenting helps to see the issue. ", "Your filtering is in the On clause not in a Where clause. ", "\nSelect ..\nFrom Table1 As tb1\n Left Join Table2 As tb2\n On tb2.Id = tb1.Id\n Left Join Table3 As tb3\n On tb3.id = tb1.id3\n And tb1.toffline = 0\n\nThus, the system is going to filter Table3 before it evaluates the Left Join to Table1. ", "If what you are attempting is to get all results where toffline is zero, then move that last filter to the Where clause:\nSelect ..\nFrom Table1 As tb1\n Left Join Table2 As tb2\n On tb2.Id = tb1.Id\n Left Join Table3 As tb3\n On tb3.id = tb1.id3\nWhere tb1.toffline = 0\n\nEDIT Noticed that toffline is from Table1.", "\n\n" ]

{ "pile_set_name": "StackExchange" }

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[ "Saturday, July 15, 2017\n\nSomething Different: Banana Split On A Stick\n\nHere is what you'll need!", "\n\nBanana Split On A Stick\n\nServings: 12\n\nINGREDIENTS\n\n4 bananas\n\n1 cup of chopped nuts\n\n1 cup chocolate, melted\n\nWhipped cream\n\nMarchino cherries\n\nPREPARATION\n1. ", "Slice a banana into thirds, and then cut each segment in half lengthwise.", "\n2. ", "On a parchment paper-lined sheet tray, place a tablespoon of chopped pecans in a small circle shape.", "\n3. ", "Pour a tablespoon of melted chocolate over the top of the nuts. ", "Lay a popsicle stick on the chocolate, and top the stick with additional melted chocolate.", "\n4. ", "Add two halves of sliced banana facing upwards in the melted chocolate. ", "Repeat with all the other banana splits on a stick.", "\n5. ", "Freeze for at least 2 hours.", "\n6. ", "When frozen solid, add a dollop of whipped cream, and top with a cherry.", "\n7. ", "Enjoy!", "\n\n8. *", "This recipe can be customized however you’d like. ", "You can choose different types of nuts, or chocolate, add sprinkles or even add ice cream if you’d like." ]

{ "pile_set_name": "Pile-CC" }

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[ "Tremor in Parkinson's disease.", "\nRest tremor is a common feature of Parkinson's disease, but its underlying pathophysiology remains unknown. ", "This review hypothesizes that tremor is related to selective loss of components of the substantia nigra. ", "The relative scarcity of tremor in related Parkinsonian conditions may indicate a dissociation associated with different pathological involvement of the substantia nigra and its connections. ", "Connections of the subthalamic nucleus with the pallidum, modified by cortical and nigral inputs, allow for the transfer of tremorogenic activity to the thalamus. ", "Thalamo-cortical interactions, tempered by cerebellar input, generate the final common pathway for tremor production." ]

{ "pile_set_name": "PubMed Abstracts" }

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[ "The United States revealed on Friday that the government conducted medical experiments in the 1940s in which doctors infected soldiers, prisoners and mental patients in Guatemala with syphilis and other sexually transmitted diseases.", "\n\nThe experiments, led by a federal doctor who helped conduct the famous Tuskegee syphilis study in Alabama, involved about 1,500 men and women who were unwittingly drafted into studies aimed at determining the effectiveness of penicillin.", "\n\nThe tests, which were carried out between 1946 and 1948, infected subjects by bringing them prostitutes who were either already infected or purposefully infected by the researchers and by using needles to open wounds that could be contaminated.", "\n\n\"Although these events occurred more than 64 years ago, we are outraged that such reprehensible research could have occurred under the guise of public health,\" Secretary of State Hillary Rodham Clinton and Health and Human Services Secretary Kathleen Sebelius said in a joint statement apologizing for the experiments. \"", "We deeply regret that it happened, and we apologize to all the individuals who were affected by such abhorrent research practices.\"", "\n\nPresident Obama had been briefed about the revelations and called Guatemalan President Alvaro Colom to \"personally express that apology,\" White House spokesman Robert Gibbs said.", "\"Obviously, this is shocking. ", "It's tragic. ", "It's reprehensible,\" Gibbs said.", "\n\nThe Guatemalan government planned to investigate, saying it \"deeply deplores that these experiments affected innocent people,\" according to a statement issued late in the day.", "\n\nIn addition to exposing another episode of unethical medical experimentation, officials said the revelations were concerning because they could further discourage already often-suspicious minorities and others from participating in medical research. ", "They also come as U.S. drug companies are increasingly going to poor, less-educated countries to test new drugs and other therapies.", "\n\n\"At a time when so much medical research is global, it behooves us to take account of what has been done in the past by American researchers in other countries,\" said Susan M. Reverby, a professor in the history of ideas and professor of women's and gender studies at Wellesley College in Massachusetts who discovered the experiments while investigating Tuskegee for a book.", "\n\nIn Tuskegee, perhaps the most notorious medical experiment in U.S. history, hundreds of African American men with late-stage syphilis were left untreated to study the disease between 1932 and 1972. ", "In the Guatemala case, the subjects were treated, but it remains unclear whether they were treated adequately or what became of them.", "\n\nReverby discovered the experiments while reading papers in the University of Pittsburgh's archives from John C. Cutler, a doctor with the federal government's Public Health Service who later participated in Tuskegee. ", "He died in 2003.", "\n\n\"I almost fell out of my chair when I started reading this,\" Reverby said in a telephone interview. \"", "Can you imagine? ", "I couldn't believe it.\"", "\n\nThe studies were sponsored by the Public Health Service, the National Institutes of Health and the Pan American Health Sanitary Bureau (now the World Health Organization's Pan American Health Organization) and the Guatemalan government. ", "The goal was to assess whether taking penicillin right after sex would prevent sexually transmitted infections." ]

{ "pile_set_name": "OpenWebText2" }

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[ "A server is a central computer that serves computers in a network environment and provides necessary functionalities to these network computers such as storage, processing, and exchange of information. ", "Conventional servers may be implemented similarly to conventional personal computers and generally comprise a central processing unit(s) (CPU), a memory(ies) as well as input/output device(s) which are all communicatively coupled together internally via a bus. ", "These internal components of the server or server hardware operate according to inherent specifications and may be influenced by external factors such as temperature, humidity, pressure and the like.", "\nA single server may be composed of a plurality of server racks which accommodate the afore-mentioned server hardware. ", "These server racks are generally arranged or stacked one over another in a compact manner and forming a server cabinet in order to minimize the room occupied by the server. ", "The so-arranged server cabinet may be disposed in a factory, an engine room or any other location suitable for physical storage of the server cabinet. ", "When more than one server cabinet is required for implementing a server, the more than one server cabinet are usually disposed one next to the other so as to minimize the room occupied by the server.", "\nDue to the compact stacking or arrangement of the server racks and of the server cabinets, some of the components of the server may be difficult to access during maintenance or repair of the server. ", "Additionally, the proximity of internal components may have an effect on the performance of the server due to heat generation and its influence on inherent specifications of the internal components." ]

{ "pile_set_name": "USPTO Backgrounds" }

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[ "Providers for Most Dental Insurance\n\nCall Us for All Your Dental Needs!", "\n\nTrusted Dentists in Palmdale\n\nFor Appointments, Call\n\nQuality Dental Care Services in Palmdale\n\nPacific Dental Care offers exceptional general and cosmetic dentistry services for patients throughout the Palmdale area. ", "We are committed to providing comprehensive dental care services and help you achieve a beautiful smile.", "\n\nWhether you need crowns or dental implants, you can rely on us. ", "Pacific Dental Care specializes in offering dental care in a comfortable environment.", "\n\nOur dentists have the required expertise to diagnose your oral problems and suggest the most appropriate treatment." ]

{ "pile_set_name": "Pile-CC" }

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[ "Synthesis of precipitating chromogenic/fluorogenic β-glucosidase/β-galactosidase substrates by a new method and their application in the visual detection of foodborne pathogenic bacteria.", "\nWe developed a new efficient method for the synthesis of important indoxyl glycoside substrates for β-glucosidase and β-galactosidase by using 1-acetylindol-3-ones as intermediates. ", "This method was used to synthesise novel precipitating fluorogenic substrates for β-glucosidase based on 2-(benzothiazol-2'-yl)-phenols. ", "We also assessed the application of these substrates in the detection of foodborne pathogenic bacteria." ]

{ "pile_set_name": "PubMed Abstracts" }

[ 0.0006998615572229028, 0.0008515294175595045, 0.0013505244860425591, 0.0005326231475919485 ]

[ "The present invention is directed to an asymmetrical differential amplifier as a level converter having a first and a second transistor, a constant current source and first and second resistors. ", "A first terminal of the first transistor and a first terminal of the second transistor are connected in common to a first circuit terminal via the constant current source and a second terminal of the first transistor is connected to a second circuit terminal via the first resistor and a second terminal of the second transistor is connected to the second circuit terminal via the second resistor. ", "A first input is connected to a control terminal of the first transistor, a second input (V.sub.ref) is connected to a control terminal of the second transistor and a first output (A) is connected to a second terminal of the first transistor and a second output (A) is connected to the second terminal of the second transistor.", "\nA differential amplifier is a symmetrical DC amplifier having two inputs and two outputs. ", "It is usually constructed with two transistors, whereby one terminal of each of the two transistors is connected to a constant current source. ", "The constant current source provides that the sum of the currents through both transistors remains constant. ", "When a voltage differing in amplitude is applied to the two inputs of the differential amplifier, then the current division in the differential amplifier varies. ", "The current through one transistor increases whereas the current through the other transistor decreases. ", "The sum of the two current magnitudes, however, is equal to the current magnitude that is supplied by the constant current source. ", "An input voltage difference produces an output voltage change in a differential amplifier. ", "A temperature-conditioned change of the voltages at the transistors acts like a common mode modulation and remains ineffective. ", "A differential amplifier is therefore preferably also used as a DC amplifier.", "\nBasic circuits for differential amplifiers have been known from the prior art literature for a long time. ", "The book \"Halbleiter-Schaltungstechnik\" by Tietze-Schenk, Fifth revised edition, page 59, discloses the basic circuitry of a differential amplifier and the following pages 59-64 show the structure and the functioning of a differential amplifier composed of npn bipolar transistors. ", "In contrast thereto, page 89 and FIG. ", "5.14 in the same reference show a basic circuit of a field effect differential amplifier, whereby this is constructed with two n-channel barrier [depletion] layer field effect transistors.", "\nSignificantly better results with respect to the input current, the bandwidth and the noise can be achieved by using field effect transistors in comparison to the circuit in FIG. ", "4.32 of the reference. ", "Further particulars regarding field effect differential amplifiers may be found on pages 90-92 in the reference.", "\nThe realization of a differential amplifier either with bipolar transistors or with field effect transistors determines the advantages and disadvantages that are critical for the respective transistor form. ", "The realization of a differential amplifier with bipolar transistors or in ECL technology has the disadvantage that the transistors must be insulated from one another. ", "For a differential amplifier in this technology, very low logical output levels as a result of low currents yield a relatively high processing speed. ", "The logical output levels usually lie in the range from 0 through 200 millivolts. ", "Relatively high logical output levels result for a differential amplifier with field effect transistors, for example, using CMOS technique; however, such a circuit is slower in comparison to a differential amplifier that uses bipolar transistors. ", "A relatively high packing density of the individual transistors is possible with the use of CMOS circuit technology. ", "Logical output levels here lie between 0 and 5 volts." ]

{ "pile_set_name": "USPTO Backgrounds" }

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[ "What Trump is really doing in Poland\n\nPresident Trump touched down in Poland on Thursday ahead of G20 talks in Hamburg, Germany.", "\n\nOn arrival he told the gathered crowds; ‘The fundamental question of our time is whether the West has the will to survive.’", "\n\nFor investors, particularly climate conscious ones, a more fundamental question is how this stop-over in Europe’s biggest coal-dependent country will impact co-ordinated efforts to decarbonize the global economy.", "\n\nOver the past few months demand for emerging markets has been on the rise, with many investors and flows going toward Asia-focused funds. ", "We asked fund selectors whether they are looking beyond their local comfort zones and investing in the Asian market.", "\n\nWe use cookies to give you the best experience on our website. ", "You can continue to use the website and we'll assume that you are happy to receive cookies. ", "If you would like to, you can find out more about cookies and managing them at any time here. ", "This site is for Professional Investors only, please read our Risk Disclosure Notice for Citywire’s general investment warnings\n\nWe use cookies to improve your experience. ", "By your continued use of this site you accept such use. ", "To change your settings please see our policy." ]

{ "pile_set_name": "Pile-CC" }

[ 0.000533055339474231, 0.0007152872858569026, 0.0005877911462448537, 0.000540733162779361, 0.0005420485395006835, 0.0005057741655036807, 0.0005280108889564872, 0.0005339810741133988, 0.0005589381325989962, 0.0006372389034368098, 0.000545635586604476 ]

[ "Introduction {#Sec1}\n============\n\nSelf-assembly is the most effective route to the creation of complex nanoscale components and, by exploiting specific non-covalent interactions, highly ordered structures can be formed (Fletcher et al. [", "@CR15]; Yu et al. [", "@CR63]). ", "In some cases these supramolecular assemblies need to be integrated into microscale sensors and actuators. ", "In this paper we address the creation of a highly efficient immunosensor by integration of top down manufacture and bottom up self-assembly of engineered proteins. ", "Whilst the surface immobilisation of antibodies provides the opportunity to create biosensors and other targeted nanoscale diagnostic devices, the immobilised components can be either randomly arranged or oriented for optimal antigen capture (Trilling et al. [", "@CR57]). ", "The simplest way to immobilise an antibody is through physical adsorption to a surface such as silicon. ", "In the case of adsorption of immunoglobulin G (IgG) to silica surfaces, the IgG will adsorb with a flat-on orientation with some overlap of IgG molecules at higher surface coverage (Xu et al. [", "@CR59]). ", "With physical adsorption of IgG, antigen can still be bound to the antibody, even with an albumin blocking agent co-adsorbed (Cowsill et al. [", "@CR11]). ", "However, the theoretical maximum binding ratio is rarely achieved, particularly at high antibody surface coverage (Xu et al. [", "@CR60], [@CR61]). ", "On the other hand, oriented antibody immobilisation enhances antigen binding as all the binding regions face away from the surface, increasing the potential to capture antigen (Huy et al. [", "@CR24]; Le Brun et al. [", "@CR33]; Liu et al. [", "@CR35]; Shen et al. [", "@CR50]). ", "Orientation thus increases the signal per unit area allowing the device size to be minimised. ", "Oriented immobilisation is usually *via* an intermediate protein immobilised to the surface which binds the antibody through a domain outside the antigen binding region (Trilling et al. [", "@CR57]). ", "Oriented binding has also been achieved by other methods such as immobilisation to a surface *via* the polysaccharide chain on IgG (Ho et al. [", "@CR22]).", "\n\nWhen creating biomimetic surfaces to integrate into microelectronics the surface that the biomolecules are to be immobilised to has to be considered. ", "Gold is an attractive option as the surface is stable, conductive (Cornell et al. [", "@CR10]) and amenable to label free techniques such as surface plasmon resonance (SPR) (Skottrup et al. [", "@CR52]), surface acoustic waves (SAW) (Kogai et al. [", "@CR31]), and quartz crystal microbalance with dissipation (QCM-D) (Amano and Cheng [@CR3]) providing a rapid and sensitive form of detection. ", "Gold also allows for self-assembly, the spontaneous deposition of a defined molecular layer by chemisorption. ", "Molecules that self-assemble usually have a chemical group that will interact strongly with the surface allowing for the assembling molecules to orient themselves in a particular direction. ", "Due to the strong interaction between the assembling molecule and the surface all the surface available for assembly becomes occupied promoting the formation of a dense monolayer. ", "The chemistry for creating self-assembled monolayers on gold is well understood and relatively straightforward (Dubois and Nuzzo [@CR12]; Love et al. [", "@CR36]). ", "Thiol-alkanes with functional groups at the alkyl terminus can be adsorbed onto gold surfaces through the thiol group adsorbing to the gold surface to form self-assembled monolayers. ", "For example, polyethylene glycol terminated thiol-alkanes have be used to create surfaces that are resistant to protein adsorption (Prime and Whitesides [@CR42]).", "\n\nPrevious publications have described the development of systems where outer membrane proteins from *Escherichia coli* can be used as engineered protein scaffolds on solid supports (Cisneros et al. [", "@CR8]; Holt et al. [", "@CR23]; Keegan et al. [", "@CR29]; Terrettaz et al. [", "@CR56]). ", "The transmembrane section (residues 1 to 171) of outer membrane protein A (tmOmpA) has been engineered such that it can be immobilised to gold surfaces in a specific orientation through a single cysteine residue (Shah et al. [", "@CR49]). ", "The tmOmpA was then circularly permutated in extracellular loop 1 to create ctOmpA which acts as a scaffold to which functional domains from other proteins can be fused (Le Brun et al. [", "@CR33]). ", "For example the ctOmpA scaffold has been fused to a single or double immunoglobulin-binding Z domain (ZctOmpA & ZZctOmpA) from *Staphylococcus aureus* protein A for antibody binding and was used to build arrays with bound IgG for antigen sensing (Le Brun et al. [", "@CR32], [@CR33], [@CR34]). ", "However, whilst the ZZctOmpA arrays bound rabbit IgG tightly, they worked poorly for mouse monoclonal IgG which are the preferred component of many medical tests (Le Brun et al. [", "@CR33]).", "\n\nProtein G from *Streptococcus* group C or G is a multi-domain cell wall-associated protein that binds IgG and human serum albumin. ", "The B domain (herein referred to as protein G), which binds IgG, has an architecture consisting of a single α-helix packed into a three-stranded β-sheet (Gallagher et al. [", "@CR16]). ", "Whilst protein G and protein A have no sequence or structural homology (Tashiro and Montelione [@CR55]), both compete for the same binding site on the constant (Fc) region of IgG, leaving the variable antigen-binding (Fab) region free to bind antigen (Sauer-Eriksson et al. [", "@CR48]). ", "Despite proteins A and G competing for the same Fc binding site, protein G binds more subclasses of IgG (Akerström et al. [", "@CR2]; Björck and Kronvall [@CR7]). ", "This is likely to arise from the difference in the protein-protein interaction between protein G and protein A to Fc. ", "The protein A-Fc interaction is mainly between hydrophobic residues whereas that of protein G mainly consists of salt bridges and hydrogen bonds between polar residues (Sauer-Eriksson et al. [", "@CR48]).", "\n\nProtein G and Protein A have been used as IgG capture molecules in immunosensor devices fabricated using self-assembled monolayers (Makaraviciute and Ramanaviciene [@CR38]; Trilling et al. [", "@CR57]). ", "The IgG capture molecules have been immobilised to surfaces using self-assembled monolayers of thiolalkanes functionalised with carboxylic acid groups for amine cross-linking (Jyoung et al. [", "@CR27]), nitrolotriacetic acid (NTA) for binding proteins with a hexa-histidine tag (Roth et al. [", "@CR43]), or for binding biotinlyated proteins (Jung et al. [", "@CR26]). ", "Monolayers of proteins A and G have been formed by direct assembly onto gold by incorporating thiol groups (Oh et al. [", "@CR40]), or through the use of fusion proteins such as glutathione *S*-transferase (GST) (Ha et al. [", "@CR18]) or elastin (Gao et al. [", "@CR17]). ", "This work describes an array of ctOmpA protein G fusions designed to reduce the dissociation of mouse monoclonal IgG. Two protein G domains were fused to the N-terminus of ZctOmpA to create the array protein GGZctOmpA. Once the GGZctOmpA is immobilised to the gold surface the monolayer is completed with a thiol-terminated amphiphile with a poly(ethylglycol) head group. ", "This amphiphilic filling molecule supports the array protein and reduces subsequent non-specific binding (Cisneros et al. [", "@CR8]; Le Brun et al. [", "@CR33]; Terrettaz et al. [", "@CR56]). ", "The GGZctOmpA array is designed for the label free detection of protein analytes in clinical or veterinary samples and when loaded with a monoclonal IgG the array becomes highly specific for the binding of defined epitopes of the target antigen. ", "This specificity in turn assembles oriented antigen allowing a second different monoclonal antibody to target another epitope on the other face of the antigen allowing a \"sandwich assay\" of high sensitivity to be created. ", "The target antigen in this work is the influenza nucleoprotein (NP). ", "NP forms oligomers that encapsulate the genomic RNA and the RNA polymerase of the influenza virus in ribonucleoprotein particles (Coloma et al. [", "@CR9]; Ruigrok et al. [", "@CR44]; Ye et al. [", "@CR62]). ", "The nucleoprotein, highly conserved between strains and type specific for influenza A and influenza B, has been a preferred target protein for influenza detection over the highly variable surface glycoproteins haemagglutinin and neuraminidase (Farris et al. [", "@CR14]). ", "This paper describes the *in situ* physical characterisation of the entire assembly process using surface plasmon resonance, quartz crystal microbalance with dissipation and neutron reflectometry.", "\n\nMaterials and methods {#Sec2}\n=====================\n\nMaterials {#Sec3}\n---------\n\nAntibodies were purchased from HyTest Ltd (Turku, Finland). ", "Molecular biology and protein purification materials were purchased from Invitrogen, Novagen and GE Healthcare (UK). ", "D~2~O was purchased from CDN Isotopes (Canada). ", "Cross-linking reagents were purchased from Pierce Scientific (Cramlington, UK). ", "All other materials were purchased from Sigma-Aldrich unless otherwise stated.", "\n\nConstruction of expression plasmids {#Sec4}\n-----------------------------------\n\nDNA digestions, ligations and transformations were carried out using standard molecular biology techniques (Sambrook and Russell [@CR46]). ", "The GGZctOmpA protein was constructed by designing a gene encoding a tandem pair of B-domains from protein G and ligating this to a scaffold protein (ZctOmpA) that contains a single N-terminal Z-domain fused to a circularly permuted tmOmpA scaffold (Le Brun et al. [", "@CR33]). ", "This construct also contains a His~6~-tag at the N-terminus.", "\n\nThe amino acid sequence of influenza A nucleoprotein (NP) was obtained from the NCBI Influenza Resource website (Bao et al. [", "@CR4]), reverse translated and codon optimised to provide a DNA sequence for expression in *E. coli*. ", "The gene was synthesised by Epoch Biolabs (Texas, USA) and subsequently ligated into a pET3d-based expression vector that incorporates a His~6~-tag at the N-terminus of the expressed protein. ", "This protein is referred to as recombinant NP (rNP).", "\n\nProtein expression and purification {#Sec5}\n-----------------------------------\n\n### GGZctOmpA {#Sec6}\n\nGGZctOmpA was expressed and purified as described previously for ZZctOmpA (Le Brun et al. [", "@CR33]) and a detailed description is included in the supplementary information. ", "Refolding of the GGZctOmpA protein was confirmed by Circular Dichroism spectroscopy (Supplemental Fig.", " [S1](#MOESM1){ref-type=\"media\"}).", "\n\n### Recombinant influenza A NP protein {#Sec7}\n\nExpression of rNP was carried out in *E. coli* BL21-AI cells in LB broth and was induced with 0.2 % L-arabinose. ", "The rNP was purified from the cell lysate by immobilised metal affinity chromatography followed by cation exchange chromatography. ", "The purified protein was analysed by SDS PAGE to ensure \\>95 % homogeneity of rNP (\\~57 kDa). ", "The binding of anti-Influenza A NP mouse monoclonal IgGs mAb245 and mAbA108 (HyTest) was confirmed by Western blotting (data not shown). ", "A detailed description of the expression and purification of rNP can be found in the supplementary information.", "\n\n### Deuterated proteins {#Sec8}\n\nDeuterated proteins were produced as described for GGZctOmpA but the media used was Silantes OD ^2^H rich growth media (Silantes, Munich, Germany) which is a rich media containing D~2~O for deuterium incorporation into proteins. ", "Mass spectrometry analysis to determine the level of deuteration was carried out by the Sydney University Proteome Research Unit. ", "A tryptic digest of protein from a gel slice of an SDS-PAGE band corresponding to the molecular weight of the protein was analysed by MALDI-TOF mass spectrometry. ", "The amount of deuteration was determined by comparing the mass of the peaks from each fragment of the hydrogenous protein with that of the corresponding peaks in the deuterated protein (Supplementary Table [S1](#MOESM1){ref-type=\"media\"}).", "\n\nSurface plasmon resonance (SPR) {#Sec9}\n-------------------------------\n\nSPR experiments used BIAcore Au sensor chips in a BIAcore X100. ", "The Au chip was cleaned by using piranha solution (3:1 H~2~SO~4~/H~2~O~2~ by volume) on the gold surface for 15 min followed by copious washing with 2 % Hellmanex (Hellma, UK) and ethanol with \\>18 MΩ water washes between each step. ", "The sensor was immersed in a 1 % (*v/v*) β-mercaptoethanol (BME) in ethanol solution to passivate the gold surface (Terrettaz et al. [", "@CR56]), washed with 1 % (*w/v*) SDS followed by \\>18 MΩ water. ", "The running buffer for all experiments was phosphate buffered saline (PBS, 20 mM sodium phosphate pH 7.6, 137 mM NaCl and 2.7 mM KCl). ", "The volumes and flow rates used for the assembly of the GGZctOmpA array; binding and crosslinking of antibody; and binding of antigen and secondary antibody are outlined in Supplementary Tables [S2](#MOESM1){ref-type=\"media\"}, [S3](#MOESM1){ref-type=\"media\"} and [S4](#MOESM1){ref-type=\"media\"} respectively with experiments repeated in triplicate.", "\n\nQuartz crystal microbalance with dissipation (QCM-D) {#Sec10}\n----------------------------------------------------\n\nThe QsenseE4 instrument (Q-Sense, Gothenburg, Sweden) with a peristaltic pump (Ismatec SA, Glattbrugg, Switzerland) used a flow rate of 50 μL min^−1^, a constant temperature of 24 °C and gold-coated sensor crystals (QSX-301, Q-sense). ", "The sensor crystals were cleaned and prepared with BME in the same way as for SPR gold surfaces. ", "The change in frequency (Δf) was measured at its fundamental frequency (5 MHz) of the quartz crystal and for the third, fifth, seventh, ninth and eleventh overtones of the fundamental frequency. ", "Data was processed into frequency and dissipation *vs.* time with four individual experiments carried out to test for repeatability. ", "A decrease in frequency corresponds to an increased mass on the surface of the sensor. ", "For each overtone the change in dissipation (ΔD) was also measured. ", "The dissipation is the proportion of energy dissipated during one cycle of the frequency oscillation and provides information on the viscoelastic properties of the materials deposited on the sensor surface. ", "Further details can be found in the supplementary information.", "\n\nNeutron reflectometry (NR) {#Sec11}\n--------------------------\n\nNR experiments were carried out on the Platypus time-of-flight neutron reflectometer (James et al. [", "@CR25]; Saerbeck et al. [", "@CR45]) at the 20 MW OPAL research reactor (Sydney, Australia) and on the Polref instrument (Webster et al. [", "@CR58]) at the ISIS pulsed neutron source (Didcot, UK). ", "Both reflectometers were used in polarised mode to enable magnetic contrast neutron reflectometry (Holt et al. [", "@CR23]; Kirby et al. [", "@CR30]). ", "The collected data was normalised to direct beam runs collected under identical conditions. ", "Absolutely scaled data was analysed using the MOTOFIT reflectivity analysis software (Nelson [@CR39]). ", "A least squares fitting routine, which selects the best fit by minimising *χ*^2^ values between model and experimental data by varying the thickness, interfacial roughness and neutron scattering length density (nSLD) of each layer, was utilised. ", "Error analysis was carried out using a Monte Carlo resampling procedure (Heinrich et al. [", "@CR21]; Holt et al. [", "@CR23]) (see supplementary information).", "\n\nResults {#Sec12}\n=======\n\nAssembly of the GGZctOmpA array {#Sec13}\n-------------------------------\n\nThe assembly of the GGZctOmpA array was carried out using established methods on a gold surface passivated with BME to improve protein assembly and maintain structure (Cisneros et al. [", "@CR8]; Holt et al. [", "@CR23]; Keegan et al. [", "@CR29]; Terrettaz et al. [", "@CR56]). ", "The GGZctOmpA was incubated on the gold surface and this was followed with an incubation of a lipid mimic of 1-mercaptoundecyl-11-triethylene glycol (thioPEG) which supports the GGZctOmpA and reduces non-specific binding in subsequent steps. ", "Each incubation step is followed with a 1 % SDS wash to remove non-specifically bound material and a PBS wash to remove excess SDS (Supplementary Figs.", " [S2](#MOESM1){ref-type=\"media\"} and [S3](#MOESM1){ref-type=\"media\"}). ", "QCM-D showed that 1.36 ± 0.04 mg m^−2^ of GGZctOmpA remained bound on the surface after a single 20 min incubation of protein solution followed by SDS/PBS wash steps (Supplementary Fig.", " [S3](#MOESM1){ref-type=\"media\"}). ", "SPR analysis of the assembly process showed that a higher surface coverage can be achieved with multiple incubation/wash steps with the gold surface saturated with GGZctOmpA at approximately 3550 RU after three incubation/wash steps (Supplementary Fig.", " [S2](#MOESM1){ref-type=\"media\"}). ", "This corresponds to 3.55 mg m^−2^ of protein on the surface (assuming 1 RU = 0.001 mg m^−2^ (Stenberg et al. [", "@CR54])) and agrees with previous published data on the assembly of ZZctOmpA (Le Brun et al. [", "@CR33]). ", "The QCM-D shows low values of ΔD and all overtones overlapping (Supplementary Fig.", " [S3](#MOESM1){ref-type=\"media\"}a, red trace) indicating that the monolayer is complete with little solvent content and is rigid.", "\n\nNeutron reflectometry provides real space structural data for surfaces and interfaces along the axis perpendicular to the lateral plane of the surface. ", "The layer structure consisted of the silicon wafer followed by the Permalloy layer that acts as a binder between the silicon and the gold layer then finally the array which is assembled on the gold surface. ", "Based on the results from the SPR experiments three protein incubations and SDS/buffer washes were used to assemble GGZctOmpA on gold surfaces for NR measurements. ", "A measurement of GGZctOmpA only on the gold surface was not carried out using NR as it has been previously shown that array proteins based on the ctOmpA scaffold can adopt many orientations on the surface unless supported by a monolayer of thioPEG (Le Brun et al. [", "@CR33]). ", "For best fitting of the NR data the array layer had to be split into two discrete layers: one layer of the thioPEG plus GGZctOmpA (plus solvent) which was 22 ± 2 Å thick and a second layer of GGZctOmpA and solvent only at 113 ± 16 Å (Fig.", " [1](#Fig1){ref-type=\"fig\"}). ", "This made a total thickness of 135 ± 18 Å for the GGZctOmpA (Fig.", " [1](#Fig1){ref-type=\"fig\"}) which is consistent with similar array proteins such as ZZctOmpA at 128 Å (Le Brun et al. [", "@CR33]). ", "The neutron scattering length density (nSLD) can be considered the neutron's refractive index and is sensitive to the isotopic composition and physical density of each layer. ", "By comparing theoretical nSLD values with those obtained from the fitted nSLD a volume fraction for each component can be derived. ", "A deuterated version of the GGZctOmpA was produced (Supplementary Table [S1](#MOESM1){ref-type=\"media\"}) as the difference in scattering length between hydrogen and its stable isotope deuterium will allow a deuterated GGZctOmpA to be distinguished between hydrogenous thioPEG and IgG. Fig.", " [1](#Fig1){ref-type=\"fig\"} shows the NR data from the array in a H~2~O contrast and the 113 Å layer with a higher nSLD then that of the thioPEG containing layer and H~2~O as expected. ", "Analysis of the nSLD values of the thioPEG plus GGZctOmpA layer shows that there is some solvent associated with the layer as there is a D~2~O/H~2~O dependent change in the nSLD and the volume fraction of solvent is 0.143 ± 0.019 (Table [1](#Tab1){ref-type=\"table\"}). ", "This corresponds with the low dissipation values seen in the QCM-D trace once the array is complete (Supplementary Fig.", " [S3a](#MOESM1){ref-type=\"media\"}). ", "From the nSLD of the GGZctOmpA only layer the volume fraction of the d-GGZctOmpA is 0.224 ± 0.018 and this corresponds to a surface coverage of 0.93 ± 0.20 mg m^−2^, which is comparable to the values observed for QCM-D although lower than the SPR results.", "Fig. ", "1**a** Magnetic contrast neutron reflectivity (points with *error bars*) and fit (*solid lines*) of an array of deuterated GGZctOmpA surrounded with thioPEG in a H~2~O contrast. **", "b** The corresponding real-space nSLD profile with zero distance set as the interface between gold on the array. ", "The gold layer has been truncated to show Permalloy layer and its different nSLD between spin up (*red*) and spin down (*black*) neutrons, and the array layers (with an illustration in the background to depict the different array layers) in detailTable 1The fitted properties of the completed array from neutron reflectometryLayerThickness / ÅnSLD / ×10^−6^ Å^−2^Volume fractionRoughness / ÅPermalloy43 ± 17.36 ± 0.10 (down)--411.06 ± 0.13 (up)Gold231 ± 34.50^a^--4thioPEG + d-GGZctOmpA22 ± 21.19 ± 0.16 (D~2~O)0.633 ± 0.085 (thioPEG only)50.20 ± 0.04 (H~2~O)d-GGZctOmpA only113 ± 16^b^0.64 ± 0.05 (H~2~O)0.224 ± 0.018518 ± 3^c^mAb245 only135 ± 116.33 ± 0.05 (D~2~O)0.067 ± 0.0075−0.45 ± 0.09 (H~2~O)mAb245 + rNP201 ± 226.19 ± 0.04 (D~2~O)0.095 ± 0.015 (rNP only)50.40 ± 0.12 (H~2~O)SubphaseN/A6.35 (D~2~O)^a^--18−0.56 (H~2~O)^aa^nSLD values were fixed at the values quoted in fits^b^thickness value before antibody addition^c^thickness when mAb245 is bound\n\nAntibody and antigen binding {#Sec14}\n----------------------------\n\nBinding of mouse monoclonal anti-influenza type A IgG2b (mAb245) to the array was measured by SPR and QCM-D (Supplemental Figs.", " [S4](#MOESM1){ref-type=\"media\"} and [S5](#MOESM1){ref-type=\"media\"}). ", "The binding of mAb245 over a range of concentrations was investigated using SPR and showed saturation of the array at high concentrations (Supplemental Fig.", " [S4](#MOESM1){ref-type=\"media\"}). ", "Figure [2](#Fig2){ref-type=\"fig\"} and Supplemental Fig.", " [S4](#MOESM1){ref-type=\"media\"} demonstrate that the mAb remains tightly bound to the array after injection, evidenced by the minimal dissociation observed. ", "A high surface coverage of GGZctOmpA was used to best reflect a real device and gain the maximum amount of mAb binding, therefore the k~on~ and k~off~ (and hence the K~d~) cannot be accurately determined between individual IgG and GGZctOmpA molecules due to mass transport and avidity affects. ", "QCM-D experiments were carried out to complement the SPR data and found that typically 1.50 ± 0.07 mg m^−2^ of mAb245 bound to the array (Supplemental Fig.", " [S5](#MOESM1){ref-type=\"media\"}). ", "The QCM-D data further confirmed slow dissociation from the array (Supplemental Fig.", " [S5](#MOESM1){ref-type=\"media\"}, 25 to 40 min) which is an improvement compared to when the IgG capture protein is the Z domain of protein A (Le Brun et al. [", "@CR33]). ", "As shown by the increase in dissipation (Supplemental Fig.", " [S5](#MOESM1){ref-type=\"media\"}, red trace), bound mAb245 is non-rigid and no longer obeys the Sauerbrey equation. ", "The array was measured by NR with mAb bound, using both spin up and spin down neutrons as well as using a D~2~O and H~2~O contrast (Fig.", " [3a](#Fig3){ref-type=\"fig\"}). ", "The real-space nSLD profile showed that the IgG layer has to be modelled as a single layer with a thickness of 135 ± 11 Å (Table [1](#Tab1){ref-type=\"table\"} and Fig.", " [3c](#Fig3){ref-type=\"fig\"}), consistent with an IgG molecule that is in the 'up right' position with the variable domain facing towards the bulk solvent. ", "The NR shows that there is a high water content in the mAb245 layer with only a 0.067 ± 0.007 volume fraction of protein. ", "This will be due to the low initial surface coverage of the GGZctOmpA and explains the high dissipation seen the QCM-D data. ", "Between the 135 Å IgG layer and the 22 Å thioPEG layer there was an 18 Å layer of high nSLD which corresponds to the deuterated GGZctOmpA only (Fig.", " [3c](#Fig3){ref-type=\"fig\"}). ", "This shows that the IgG does not sit on the protein resistant thioPEG layer.", "Fig. ", "2The binding of antibody and antigen to arrays of GGZctOmpA measured by SPR. ", "30 μg mL^−1^ mAb245 was bound to an array of GGZctOmpA, then rNP at a concentration range of 0 to 10 000 ng mL^−1^ was bound. ", "Secondary antibody, mAb108 (30 μg mL^−1^), was the final binding event before regeneration with 100 mM HCl. ", "1° - mAb245 injection; T -- buffer wash; A -- rNP injection, 2° - mAb108 injection, H -- regeneration with 100 mM HClFig. ", "3Reflectivity data (*symbols*) and fit (*solid lines*) of an array of d-GGZctOmpA filled with thioPEG and with **a** mAb245 bound only to the array and **b** is the reflectivity data of the array with mAb245 and rNP bound. ", "Both data have the same symbols and shows the up spin states (○ and □) and down spin states (□ and ◇) as well as the D~2~O (○ and □) and H~2~O (△ and ◇) contrasts (spin up H~2~O absent in **b**). ", "The data between the D~2~O and H~2~O contrasts is offset for clarity. **", "c** The corresponding real-space nSLD profiles of the array with mAb245 bound from the gold interface and d) is the corresponding real-space nSLD profile of the array from the gold interface with mAb245 and rNP bound\n\nThe antigen, rNP, binds to the array through the antigen binding domains of mAb245 and both SPR and QCM-D confirm that rNP binds to GGZctOmpA-immobilised mAb245 (Fig.", " [2](#Fig2){ref-type=\"fig\"} and Supplemental Fig.", " [S5](#MOESM1){ref-type=\"media\"}). ", "The rNP binding was tested over a concentration range of 1 to 10 000 ng mL^−1^ by SPR (Fig.", " [2](#Fig2){ref-type=\"fig\"}). ", "Bound rNP was measured under the same conditions as for bound mAb245 using NR (Fig [3b](#Fig3){ref-type=\"fig\"}). ", "The measured NR data with rNP bound show small changes in the shape of the fringes when compared to mAb245 only, suggesting that rNP has bound but there is no great structural change. ", "The modelling of the NR data showed that when bound to the array, rNP antibody layer still had to be modelled as one discrete layer (Fig.", " [3d](#Fig3){ref-type=\"fig\"}) as separating the antibody layer into two layers of antibody and antigen resulted in poorer fits. ", "On binding of rNP to the antibody layer the overall thickness of the layer increased to 201 ± 22 Å, strongly indicating that antigen is bound (Fig.", " [3d](#Fig3){ref-type=\"fig\"} and Table [1](#Tab1){ref-type=\"table\"}). ", "There is little change in the nSLD of the antibody layer after rNP addition and this is unsurprising due to the low surface coverage of mAb245. ", "However, the Monte Carlo resampling for the nSLD parameter shows a clear difference in nSLD before and after rNP addition (Supplementary Fig.", " [S6](#MOESM1){ref-type=\"media\"}) confirming rNP binding in Fig.", " [3c](#Fig3){ref-type=\"fig\"}. ", "Figure [4](#Fig4){ref-type=\"fig\"} shows the dimensions for each component in the GGZctOmpA array plus rNP and shows that the total thickness for the array is 241 ± 27 Å.Fig. ", "4The dimensions of each component of a GGZctOmpA array before and after IgG and rNP binding. ", "ctOmpA is shown in light green and is bound to the gold surface and is surrounded by thioPEG (the molecular packing is not represented accurately) with a protein A Z domain linker (*dark green*) and a tandem pair of B domains from protein G (*red*) to which there is an IgG (*blue*) bound at the Fc region. ", "Bound to the IgG Fab regions there is NP (*orange*) to which a secondary IgG can be bound (not shown)\n\nThe secondary antibody used was a mouse monoclonal anti-influenza virus type A IgG1 (mAb108) that was raised against a different epitope of the NP protein. ", "Binding mAb108 enhances the signal from bound rNP as shown by SPR (Fig.", " [2](#Fig2){ref-type=\"fig\"}). ", "Although we completely saturate the available protein G domains with IgG there is some dissociation of the primary IgG (mAb245) after injection which could result in trace amounts of free protein G domains. ", "The secondary antibody (which is also a mouse monoclonal IgG) can bind to any free protein G molecules which would result in some background signal in the 0 ng/mL rNP case. ", "The mass of secondary antibody that bound to the array was 3.69 ± 0.04 mg m^−2^ as determined by QCM-D (Supplemental Fig.", " [S5](#MOESM1){ref-type=\"media\"}). ", "This is a binding ratio of mAb108 to rNP of 1:0.65 (expected 1:1). ", "The large increase in QCM-D dissipation shows that mAb108 is non-rigid when bound (Supplemental Fig.", " [S5](#MOESM1){ref-type=\"media\"}a, red trace).", "\n\nSpecificity of rNP binding and the level of non-specific binding was tested with a series of control SPR experiments. ", "The first experiment was binding mAb245 and rNP to an array of ctOmpA where the tandem pair of protein G domains is absent and therefore cannot specifically capture IgG. Supplementary Fig.", " [S7](#MOESM1){ref-type=\"media\"} shows that neither mAb245 nor rNP can bind to an array of ctOmpA. A second experiment was carried out where rNP was bound to an GGZctOmpA with a IgG bound which is not specific for rNP. ", "Supplemental Fig.", " [S8](#MOESM1){ref-type=\"media\"} shows the binding of rNP to mouse monoclonal anti-FLAG M2 IgG1 (which is specific for the FLAG epitope) immobilised to an array of GGZctOmpA. Supplemental Fig.", " [S8](#MOESM1){ref-type=\"media\"}b is a comparison of the relative response of rNP binding to mAb245 and anti-FLAG M2 immobilised to GGZctOmpA with time zero set as the point of 1 μg mL^−1^ rNP injection. ", "The trace shows that rNP does not bind to the anti-FLAG M2 IgG1 but does bind to mAb245.", "\n\nCross-linking assays {#Sec15}\n--------------------\n\nTo further reduce the dissociation of IgG from GGZctOmpA arrays the IgG was chemically cross-linked to the protein G domains of GGZctOmpA in separate experiments by two different cross-linkers. ", "The first was Bis(sulfosuccinimidyl) suberate (BSSS) which is an *N*-hydroxysulfosuccinimidyl (NHS)-ester homobifunctional water soluble cross-linker with an eight carbon spacer arm of 11.4 Å. The second linker was Bis(succinimidyl) penta(ethylene glycol) (BS(PEG)~5~), which is a linker with a pentaethylene glycol spacer of 21.7 Å. Both cross-linkers are amine -- to -- amine cross-linkers that are ideal for cross-linking IgG which have many surface exposed lysine residues. ", "The cross-linking reagent at a final concentration of 0.5 mM in PBS buffer was incubated with the preformed IgG array and the reaction was terminated with 1.0 M Tris--HCl pH 6.8 (Fig.", " [5a](#Fig5){ref-type=\"fig\"} and Supplemental Fig.", " [S9](#MOESM1){ref-type=\"media\"}). ", "Non-cross-linked antibody was removed by washing with 100 mM HCl followed by a PBS wash. ", "The effectiveness of the cross-linking was assessed by comparing the amount of mAb245 bound to the array before the addition of the cross-linking agent to the amount bound after the HCl and PBS wash steps (the areas marked with the letter 'A' in Fig.", " [5a](#Fig5){ref-type=\"fig\"}). ", "Both cross-linking agents were found to cross-link with approximately 98 ± 1 % efficiency. ", "The effect of the cross-linking on the array's ability to bind rNP and mAb108 was tested and mAb245 cross-linked to an array of GGZctOmpA retained the capacity to bind rNP. ", "By comparing Figs.", " [2](#Fig2){ref-type=\"fig\"} and [5](#Fig5){ref-type=\"fig\"} cross-linked mAb245 binds rNP to the same extent as non-cross-linked mAb245 across all bulk rNP concentrations. ", "Furthermore, the array can be regenerated, by washing with 100 mM HCl to remove the rNP, leaving the cross-linked mAb245 in place (Fig.", " [5](#Fig5){ref-type=\"fig\"}). ", "This allows for multiple cycles of antigen binding and array regeneration without the need to replenish the array with primary IgG on each new addition of antigen. ", "Secondary antibody (mAb108) binding to antigen captured by cross-linked mAb245 was also demonstrated with SPR (Fig.", " [5b](#Fig5){ref-type=\"fig\"}).Fig. ", "5**a** Cross-linking IgG to an array of GGZctOmpA with BSSS. ", "30 μg mL^−1^ of mAb245 was incubated on the surface for 300 s at a flow rate of 5 μL min^−1^ this was followed by wash with PBS buffer. ", "An incubation of 0.5 mM BSSS for 900 s was carried out to initialise cross-linking with the reaction terminated with 1 M Tris--HCl pH 6.8 followed by a double wash of 100 mM HCl. ", "The before and after response to the crosslinking was assessed in the areas marked 'A' **b** Binding of rNP (over a concentration range of 0.1 to 10 000 ng mL^−1^) to mAb245 that has been cross-linked to an array of mAb245 with BSSS cross-linker followed by binding of mAb108 (30 μg mL^−1^) at a flow rate of 5 μL min^−1^. Between each concentration the array was regenerated with a wash of 100 mM HCl. ", "Data was subtracted from a data set of 0 ng mL^−1^ antigen\n\nDiscussion {#Sec16}\n==========\n\nThe analytical methods provide a comprehensive picture of the processes occurring during self-assembly and the combination of SPR, QCM-D and NR provides both overlap and complementarity. ", "Protein G B-domains were successfully fused to the ctOmpA scaffold with a protein A Z-domain as a linker molecule. ", "The structure of GGZctOmpA could be checked in solution using circular dichroism spectroscopy (Supplementary Fig.", " [S1](#MOESM1){ref-type=\"media\"}) before forming self-assembled monolayers on gold surfaces (Fig.", " [1](#Fig1){ref-type=\"fig\"} and Supplementary Figs.", " [S2](#MOESM1){ref-type=\"media\"} and [S3](#MOESM1){ref-type=\"media\"}) as shown for other β-barrelled scaffold proteins (Le Brun et al. [", "@CR33], [@CR34]; Shah et al. [", "@CR49]; Terrettaz et al. [", "@CR56]). ", "Specific deuteration of just GGZctOmpA allowed for this array protein to be distinguished from the hydrogenous thioPEG and IgG and clearly highlights the 18 Å layer between the thioPEG / array protein layer and the IgG layer (Fig.", " [3c](#Fig3){ref-type=\"fig\"}). ", "Although similar layers have been observed previously (Le Brun et al. [", "@CR32], [@CR33]), all components were hydrogenous so that the gap between the IgG and the protein resistant thioPEG layer was previously unseen.", "\n\nProtein G has previously been bound to surfaces and the linkers used can be split into two categories resulting in either oriented immobilisation or random immobilisation (Trilling et al. [", "@CR57]). ", "This GGZctOmpA array clearly binds IgG in an oriented manner with the neutron reflectivity data showing that the IgG layer is 135 Å thick. ", "Considering the dimensions of IgG (143 × 77 × 40 Å^3^ (Silverton et al. [", "@CR51])), this would indicate an orientation which is optimal for antigen binding where the variable, antigen-binding domains are facing away from the surface (Figs.", " [3c](#Fig3){ref-type=\"fig\"} and [4](#Fig4){ref-type=\"fig\"}). ", "Although protein G can also bind the antigen-binding Fab fragments of IgG (Tashiro and Montelione [@CR55]), isothermal titration calorimetry has shown that the K~a~ of protein G binding to Fab fragments was two orders of magnitude lower than binding to Fc fragments or whole IgG, and that binding to the Fc domain dominated (Lund et al. [", "@CR37]).", "\n\nStrong binding with little dissociation between the GGZctOmpA array and mouse monoclonal IgG2b was demonstrated (Fig.", " [2](#Fig2){ref-type=\"fig\"}). ", "Arrays of ZZctOmpA, which capture IgG using the Z domain of *Staphylococcal aureus* protein A, bound mouse monoclonal IgG2a with a K~d~ of 0.778 nM (Le Brun et al. [", "@CR33]), which also indicates strong binding. ", "However, the subclass and species of origin of the IgG has to be taken into account. ", "Unlike mouse monoclonal IgG2a, mouse monoclonal IgG2b and IgG1 bind to protein A poorly (Björck and Kronvall [@CR7]), whereas protein G binds all mouse monoclonal IgG subclasses (Björck and Kronvall [@CR7]; Eliasson et al. [", "@CR13]). ", "Therefore the advantage of using protein G over protein Z as the IgG binding domain is the ability of protein G to bind a much wider range of IgG classes than protein A (Björck and Kronvall [@CR7]). ", "Eliasson et al. ([", "@CR13]) showed that creating genetic fusions of the IgG-binding domains of protein A and protein G can have additive effects. ", "In this work a Z domain from protein A was used as a linker between ctOmpA and protein G so the possibility of some additive effect in this array exists.", "\n\nPrevious work has shown that a greater antigen binding can be achieved through oriented immobilisation of IgG to surfaces (Bergström and Mandenius [@CR6]; Kausaite-Minkstimiene et al. [", "@CR28]; Song et al. [", "@CR53]) and this work demonstrated complete saturation of the antigen binding sites on the IgG (Fig.", " [2](#Fig2){ref-type=\"fig\"}). ", "Although complete saturation was achieved, a discrete antigen layer could not be resolved in the NR data; rather an overall increase in the nSLD and thickness of the antibody layer was observed (Fig.", " [3d](#Fig3){ref-type=\"fig\"} and Supplementary Fig.", " [S6](#MOESM1){ref-type=\"media\"}). ", "This indicates that the rNP, can adopt multiple IgG-bound orientations allowed by the flexible hinge region between Fc and Fab domains which orientates Fab in different arrangements relative to the position of Fc without affecting the overall shape of the IgG (Abe et al. [", "@CR1]; Harris et al. [", "@CR19]; Saphire et al. [", "@CR47]). ", "This is reinforced by the increased dissipation in the QCM-D traces suggesting flexibility in the bound IgG structure (Supplementary Fig.", " [S5](#MOESM1){ref-type=\"media\"}). ", "In Fig.", " [4](#Fig4){ref-type=\"fig\"} we only show one IgG arrangement for clarity; however the Figure does indicate that IgG fully occupied with rNP would occupy a large molecular area and this could account for the low volume fractions observed. ", "SPR and QCM-D data shows that the IgG to antigen ratio of mAb245 to rNP binding is higher than that observed previously with an array of ZZctOmpA using rabbit polyclonal anti-HSA IgG and human serum albumin as antigen where a 25 % occupancy of the antigen binding sites was achieved (Le Brun et al. [", "@CR33]).", "\n\nThe engineering of the array *via* the chemical cross-linking of IgG to the protein G moieties prevents IgG dissociation while retaining the detection capabilities of the original array (Fig.", " [5](#Fig5){ref-type=\"fig\"}). ", "In addition crosslinking the IgG to protein G stabilises the array enabling regeneration without having to replenish primary antibody *via* a simple low pH solution wash. ", "A number of approaches have been used to cross-link protein G or protein A to IgG for enhanced stability which include using cyanamide (Bereli et al. [", "@CR5]), dimethyl pimelimidate (Bergström and Mandenius [@CR6]), and bis(sulfosuccinimidyl) linkers(Song et al. [", "@CR53]). ", "In this work bis(sulfosuccinimdyl) linkers were used due to their mild conditions for cross-linking and ease of use. ", "The functional ends of the cross-linkers were separated by different spaced cross-linkers with each successfully cross-linking the IgG and protein G. Previous cross-linking studies, have found that the orientation of the IgG is more important than the cross-linking reagent (Kausaite-Minkstimiene et al. [", "@CR28]) and this work supports this.", "\n\nPrevious work creating membrane protein -- immunoglobulin-binding domain chimeras have only been tested against IgG that have been raised against model proteins such as albumin (Le Brun et al. [", "@CR33], [@CR34]). ", "By creating the GGZctOmpA molecule a wider range of monoclonal IgG subtypes can be bound which increases the prospect for a larger range of epitopes from 'real world' analytes to be detected. ", "In this work the molecule that was chosen for proof-of-concept was recombinant influenza A nucleoprotein which is currently the basis of a number of point-of-use diagnostic tests for influenza (Hatchette [@CR20]; Peters et al. [", "@CR41]).", "\n\nConclusions {#Sec17}\n===========\n\nThis work has revealed how nanoscale components (engineered protein, alkane thiol and IgG) can self-assemble into an array onto surfaces for potential microscale devices. ", "The antigen used for the proof of concept was nucleoprotein from influenza, creating a label-free platform that is amenable to both SPR and QCM-D techniques and sensitive to the presence of flu antigens. ", "Crosslinking the IgG to the array enables the benefits of non-covalent self-assembly to be used in a stable covalent structure. ", "Probing the interfacial structure of the array by neutron reflectometry using both conventional isotope contrasts and magnetic contrasts showed a structured array with optimal IgG orientation for antigen capture.", "\n\nElectronic supplementary material\n=================================\n\n {#Sec18}\n\n###### \n\n(DOCX 266 kb)\n\nBS(PEG)~5~\n\n: Bis(succinimidyl) penta(ethylene glycol)\n\nBSSS\n\n: Bis(sulfosuccinimidyl) suberate\n\nctOmpA\n\n: Circularly permuted transmembrane domain of *E. coli* outer membrane protein A\n\nGGZctOmpA\n\n: N-terminal tandem pair of B domains from protein G with a C-terminal ctOmpA scaffold\n\nIgG\n\n: Immunoglobulin G\n\nmAb245\n\n: Mouse monoclonal IgG raised against rNP\n\nmAb108\n\n: Mouse monoclonal IgG raised against rNP (different epitope to mAb245)\n\nNP\n\n: Native nucleoprotein from influenza\n\nNR\n\n: Neutron reflectometry\n\nnSLD\n\n: Neutron scattering length density\n\nPBS\n\n: Phosphate buffered saline\n\nQCM-D\n\n: Quartz-crystal microbalance with dissipation monitoring\n\nrNP\n\n: Recombinant nucleoprotein\n\nSPR\n\n: Surface plasmon resonance\n\nthioPEG\n\n: 1-mercaptoundec-11-yltriethylene glycol\n\nFunding from Technology Strategy Board grant (TSB/100565) to Orla Protein Technologies Ltd and Newcastle University for the 'Virasens' project and Wellcome Trust Grant Number 093581. ", "The authors thank the Bragg Institute Programme Advisory Committee (proposal P1500), and the ISIS Facility Access Panel (RB121134), for the award of neutron beam time. ", "APLB acknowledges a Discovery Early Career Researcher Award from the Australian Research Council (DE140101788). ", "The authors thank Marie Gillon (ANSTO) and Helen Ridley (Newcastle University) for technical assistance.", "\n" ]

{ "pile_set_name": "PubMed Central" }

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[ "As Season 3 heads into its final stretch, a shakeup in one of Albion's top guilds threatens to shift the balance of power.", "\n\nAugust 10, 2018 at 12:29 PM by PrintsKaspian\n\nSeason 3 has been Albion Online's most competitive to date. ", "Just past the halfway point on August 4, the first- and second-place teams – Money Guild and Team Casualty – were within a few thousand points of each other, a negligible number for high-ranking teams.", "\n\nTeam Casualty have long been one of Albion's top guilds and had high hopes for Season 3. ", "Ellipses, the guild leader, discussed the season (as well as their ongoing rivalry with Crimson Imperium Reborn) in a preseason interview on the forums. ", "The Fleet and Frogue One, both strong guilds in their own right, paired well with Team Casualty, and the synergy between these guilds made their alliance, ALONE, the most powerful in Albion man-for-man.", "\n\nMoney Guild's alliance, OOPS, was able to remain in power due to the combination of their elite GvG team and strong-handed diplomacy, which saw them retain control over large swaths of the Outlands while maintaining a truce with the powerful guild Crimson Imperium Reborn and their alliance Partnership of Equals. ", "But with Team Casualty able to hold large parts of Cumbria and strategic footholds in Mercia, as well as having a legitimate shot at taking any castle on the map in hand-to-hand combat, they appeared to be in fantastic position to challenge Money Guild for the top spot.", "\n\nThen, on August 8, Team Casualty awoke to find their alliance disbanded, five of their plots dropped, among them Fecund Mound Meadow, their de facto base of operations where countless resources were lost. ", "With their formerly powerful guild in disarray and their options for new resources, weapons, and food severely depleted, Team Casualty's shot at a Season 3 championship seemed in doubt.", "\n\nSo far, Team Casualty has taken this tremendous hit in stride, immediately jumping back into the fray: on August 8, the day of their devastating loss, they founded a new alliance, POWER, and were able to successfully reclaim Moonlit Grove in Mercia.", "\n\nWhether they can continue to reclaim lost territories and resources over the coming weeks remains to be seen, but with season points doubling after the final territory reset on August 18, the Season 3 crown remains in play." ]

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[ "Come Sept. 7, when the Broncos kick off their season against the Colts, Denver will be the latest team hoping to avenge a Super Bowl loss by hoisting the Lombardi Trophy a year later.", "\n\nIt took him 13 years to worry. ", "Thirteen years, and not a team had come close to doing what Marv Fleming and the Dolphins did in 1972, when they won each and every game en route to a Super Bowl championship. ", "But in 1985, as the calendar turned to November, Fleming and his teammates fretted about the Bears. ", "Chicago won its first 12 games that fall before losing to Miami, of all teams, in late November.", "\n\nAgainst the odds: Can Seahawks buck history, repeat as champions?", "\n\nThey worried again in 1998, when John Elway’s Broncos won their first 13 games, and in 2005 when Peyton Manning’s Colts did the same. ", "They wrung their hands in 2007 when New England went undefeated until the Super Bowl, and they paced in 2009, when the Saints started 13-0, the Colts 14-0. ", "Green Bay threatened most recently, in 2011, but it too fell, in Week 15 to the Chiefs.", "\n\nWith each near miss, Fleming couldn’t help but lament the thought of his team’s perfect record someday being one of two, one of three, one of many. ", "But after a few teams came close to perfection, the retired tight end began to balk at reports of his teammates’ celebrations. ", "Despite embracing a stranger in a grocery store the night the Giants defeated the Patriots, he has always put the word out: do not pop your champagne in public. ", "Drink it in a closet, he tells teammates, because you’re celebrating your own victory, not this new team’s loss.", "\n\nThe perfect season has come to define the 1972 Dolphins, and they are fiercely proud that their record has stood alone for 42 years. ", "However, 17-0 isn’t the only mark that Miami team holds onto. ", "It also remains the last Super Bowl loser to rebound with a championship the next year -- and many of the players don’t even have a clue.", "\n\nFleming doesn’t quite believe it the first time he’s told: “No one’s done it since us?” ", "he asked. “", "Really? ", "Well that’s just another record for us, I guess.” ", "When Paul Warfield, a starting wide receiver on the 1972 team, calls, he can barely eke out a hello before asking for confirmation that no, not a single team since has won the Super Bowl the season after losing it. ", "He chuckles at the notion, another piece of history his team didn’t even know it held.", "\n\nCome Sept. 7, when the Broncos kick off their season against the Colts, Denver will be the latest team hoping to avenge a Super Bowl loss by hoisting the Lombardi Trophy a year later. ", "And after their second preseason game, a 34-0 drubbing of San Francisco, the Broncos’ odds of winning the Super Bowl moved to 5-to-1, putting them in a tie with the Seahawks for the best chance at a championship.", "\n\nKING: Why 'Hawks won't repeat | Teams that could leap into playoffs\n\nHistory begs Vegas’ pardon, though. ", "Twenty-eight of the 43 Super Bowl losers since the AFL-NFL merger in 1970 have failed to win a playoff game the following season, with 12 missing the playoffs entirely the next year. ", "Just seven of the 47 previous Super Bowl losers have returned to the big game a season later, and only two have won: the 1971 Cowboys and the Dolphins a year later. ", "In short, history says that the Broncos’ chances at a Super Bowl championship come February are close to nil.", "\n\nBut Peyton Manning and company aren’t listening. ", "In fact, they don’t even want to hear the words Super Bowl, not now. ", "Not until January.", "\n\nThis is a topic that makes the Broncos bristle, that causes the most genial of players to stiffen. ", "Don’t say the words “1972 Dolphins.” ", "Don’t dare utter “Super Bowl.” ", "Tight end Jacob Tamme, who’s come off a Super Bowl loss in Indianapolis, the Broncos’ upset by Baltimore in 2012 and now this second Super Bowl defeat, is particularly aggravated by the discussion of history, by even the hint that he’s looking much past next month.", "\n\nWes Welker's latest concussion puts NFL's head injury issue in crosshairs\n\n“I don’t think about [the Super Bowl] until one of y’all asks,” Tamme said. “", "It’s all about training camp right now, every day, getting better. ", "Look, you can’t get to your big-picture, end result if you don’t focus on every step in front of you. … ", "It’s all about everyday focus.”", "\n\nThe retired Dolphins would applaud Tamme’s approach. ", "Forty-two years later, they can cop to the bitterness of losing to Dallas 24-3, to the way they fed off that disappointment and the criticism they heard from fans and other teams. “", "They’re an old AFL team,” Warfield parrots all these years later. “", "They can’t play with the big boys.” (", "Teams from the old AFL has won just two of the first six Super Bowls, contributing to a sense that the NFL had been the far superior league.) ", "Such perspective takes time, though. ", "In the immediate aftermath of losing, to do what the Broncos are hoping to do, the good ol’ cliché of living one week at a time might just ring true.", "\n\n“You play them one at a time, and that’s hard to do, but one at a time,” Warfield said. “", "You take nothing for granted. ", "We learned that you couldn’t take things for granted.”", "\n\nSo if these Broncos have a chip on their shoulder, well, they’re not saying. ", "The Dolphins had one, certainly. ", "Defensive end Bill Stanfill, who today suffers from memory loss and is a plaintiff in the NFL concussion lawsuit, remembers feeding off the disappointment of losing more than anything else from that season. ", "But here are the Broncos of 2014, claiming Seattle is just another team, that playing them twice this fall is pretty much the same as facing the not-so-ferocious Raiders twice each year.", "\n\nDenver’s is a subdued urgency. ", "It’s unspoken, but it’s visible, in the mid-practice training camp skirmishes between offense and defense, in Manning’s 15-yard taunting penalty against Houston. ", "The Seahawks were the tougher, louder team last February, and the Broncos took note. ", "A year ago, a defense that was from the start inferior to Denver’s offense and would go on to lose many of its key players didn’t have the gall to chirp or squabble. ", "It didn’t think it was as good as its quarterback and receivers, but now Denver’s defense wants to be the team’s best unit, no matter the records its offense set a season ago.", "\n\nThat’s the thing about these Broncos. ", "Unlike the Dolphins, whom Fleming and Warfield believe were cowed by the pomp and circumstance of the Super Bowl and needed the experience of a loss to understand how to win, Denver was simply battered last season. ", "Sure, plenty of its starters were inexperienced, but the iron fist of Manning kept them focused in New York, and much of the team’s youth was a result of injuries to its starting lineup. ", "In fact, if the Broncos can stay healthy and make it to the Super Bowl, they will have 11 different starters from the group they trotted out last February, thanks to free agent acquisitions and players returning from injuries. ", "The Dolphins in 1972? ", "They made just four changes in their starting lineup between Super Bowls.", "\n\nSI's All-Breakout Team: Players on the verge of a surprising 2014 season\n\n“It [is] a motivator because we didn’t finish the Super Bowl the way we wanted to,” Broncos defensive tackle Kevin Vickerson said, “and the guys that didn’t play -- like myself and a couple of other guys -- we’re taking a little bit extra motivation [into the season] just to get back to that point and get back to that game.”", "\n\nFor any team as stacked as the Broncos, health and a dose of luck are the biggest factors in terms of winning in Arizona. ", "Already, Denver has lost starting linebacker Danny Trevathan, who fractured his leg on Aug. 12 and will miss 6-8 weeks, and receiver Wes Welker suffered his third concussion in 10 months on Aug. 23. ", "However, the team expects both players to be at full speed long before midseason, and if Denver’s injury woes can regress to the mean this season, they should be plenty capable of making another run.", "\n\n“We’re very deep,” cornerback Chris Harris, who missed the Super Bowl with a torn ACL, said. “", "That’s what it’s going to take. ", "It’s going to take all [of our players]… from the No. ", "1 to the very bottom… to be able to make it to the Super Bowl again.", "\n\n“We don’t need a reminder [of our loss]. ", "A lot of guys on this team are hungry, and a lot of guys didn’t even get to play in the Super Bowl.”", "\n\nSo if those Dolphins were motivated by losing, both the game and their focus, these Broncos have a different impetus. ", "They want to give their now-healthy teammates a chance. ", "Harris, Vickerson, defensive end Derek Wolfe, offensive tackle Ryan Clady, safety Rahim Moore, linebacker Von Miller -- the list of men who had to watch the Super Bowl from the sideline goes on and on, and each wants his moment. ", "Their teammates want it for them, and they want it for Manning, because it’s hard to know how many seasons the future Hall of Famer has left.", "\n\nThat’s why the Broncos will insist that they’re thinking about tomorrow rather than yesterday, but no further than next Sunday. ", "Feb. 1, 2015 seems about as far in the future as Jan. 14, 1973 does in the past.", "\n\n“I don’t really put too much on the past,” coach John Fox said. “", "When you start letting all that other garbage get in, all it can do is disrupt your focus. … ", "You look out the windshield, and you’ve got that little [rear-view] mirror there, but it’s way smaller than the windshield.”", "\n\nRanking NFL's most concerning QB situations ahead of the 2014 season\n\nSo if the past is irrelevant, if the string of 41 consecutive teams who couldn’t rebound from Super Bowl losses means nothing, then consider the windshield. ", "If history tells these Broncos anything, it’s to keep their eyes on the road, not the horizon. ", "They can’t listen to newly minted analyst Tony Gonzalez as he predicts a perfect season, just as they had to tune out the Seahawks’ crowing over their victory all offseason.", "\n\nAnd while Warfield, Fleming and their teammates are oblivious to the long odds of repeating, the Broncos are indignant. ", "Odds said that Manning shouldn’t return to form after neck surgery; that Harris, an undrafted rookie, shouldn’t be starting. ", "Odds said defensive tackle Sylvester Williams, who dropped out of high school to work in a factory, should never have gotten out of his home state of Missouri, that tight end Julius Thomas, a former college basketball star at a mid-major school, should be playing hoops overseas, not starring in the NFL.", "\n\n“You’re looking at a field full of guys who have defied odds,” Thomas said. “", "For us to be worried about what the odds are, forget it.”", "\n\nHe’s a little bit angry that the subject must even be broached, that anyone would believe a rebound from that loss impossible. ", "He’s a little bit angry, and when it comes to 42 years of history and the 19 to 20 games that lie ahead, a little bit of angry might be just what the Broncos need." ]

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[ "Confession: I don’t mind it when airlines lose my luggage. ", "Sure, it’s inconvenient for a day or two. ", "But at the end of it, I end up with some compensation in my back pocket, and some new clothes in my wardrobe, courtesy of the airline.", "\n\nBut I accept that not everyone feels as I do. ", "For this reason, I think it’s kinda cool that Delta has included a real-time luggage tracker in the latest version of its mobile app.", "\n\nThe feature apparently cost $50 million to build. ", "It allows you to see where your stuff is – provided that it’s at one of the 84 airports that support Delta’s new tracking tech.", "\n\nHere’s how it works. ", "All bags will get an Radio Frequency Identification (RFID) tag. ", "This allows Delta to track them in real-time using radio waves.", "\n\nScanners positioned throughout the baggage system will allow Delta to monitor where the bag is, and relay that information to the passenger.", "\n\nDelta has traditionally been one of the best airlines when it comes to handling baggage. ", "During 2012, it lost only 200,000 bags. ", "That sounds like a lot, but bear in mind it carried 98 million passengers during the same period.", "\n\nYou can try the feature on your next Delta flight by grabbing the app from Google Play and the App Store. ", "Oh, and let us know how it works for you\n\nDelta passengers can now track their bags on a map on CNN Money\n\nRead next: Hillary Clinton email saga continues as 33,000 erased emails may not be 'missing' after all" ]

{ "pile_set_name": "OpenWebText2" }

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[ "Microinjection of Escherichia coli UvrA, B, C and D proteins into fibroblasts of xeroderma pigmentosum complementation groups A and C does not result in restoration of UV-induced unscheduled DNA synthesis.", "\nThe UV-induced unscheduled DNA synthesis (UDS) in cultured human fibroblasts of repair-deficient xeroderma pigmentosum complementation groups A and C was assayed after injection of identical activities of either Uvr excinuclease (UvrA, B, C and D) from Escherichia coli or endonuclease V from phage T4. ", "Under conditions where the T4 enzyme was able to induce repair synthesis in both XP complementation groups in agreement with earlier observations (de Jonge et al., ", "1985), no effect of the UvrABCD excinuclease could be observed either when the enzymatic complex was injected into the cytoplasm, or when it was delivered directly into the nucleus. ", "In addition, no effect of the E. coli excinuclease was found on the repair ability of normal repair-proficient human fibroblasts. ", "We conclude that the UvrABCD excinuclease may not work on DNA lesions in human chromatin." ]

{ "pile_set_name": "PubMed Abstracts" }

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[ "Q:\n\nDjango Adding Comments\n\nI'm trying to add a comments section at the end of my form so users can enter comments. ", "I'm following exactly what the Django tutorial is asking: https://docs.djangoproject.com/en/dev/ref/contrib/comments/example/\nBut I keep getting this error: \nTemplateSyntaxError \nCaught AttributeError while rendering: 'ModelFormOptions' object has no attribute 'proxy'\nAnyone have any idea why? ", "\nHere's the log:\n91 {% load comments %}\n92 {% get_comment_count for form as comment_count %}\n93 <p>{{ comment_count }} comments have been posted.</p>\n94 {% render_comment_list for form %}\n95 \n96 {% get_comment_list for form as comment_list %}\n97 {% for comment in comment_list %}\n98 <p> Posted by: {{ comment.user_name }} on {{ comment.submit_date }}</p>\n99 \n100 <p>Comment: {{ comment.comment }}</p>\n101 {% endfor %}\n\nThe error occurs at line 92\n\nA:\n\nThe tag requires a model instance, not a form.", "\n\n" ]

{ "pile_set_name": "StackExchange" }

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[ "\n\n\n\n\n\n\n\n\n\nIn The\n\nCourt of Appeals\n\nSixth Appellate District of Texas at Texarkana\n\n\n______________________________\n\nNo. ", "06-02-00073-CR\n______________________________\n\n\nCHARLES CLAUDE CARLTON, Appellant\n\nV.\n\nTHE STATE OF TEXAS, Appellee\n\n\n\nOn Appeal from the 248th Judicial District Court\nHarris County, Texas\nTrial Court No. ", "873521\n\n\n\n\n\nBefore Morriss, C.J., Grant and Ross, JJ.", "\nOpinion by Justice Ross\n\nO P I N I O N\n\n\tCharles Claude Carlton appeals from his conviction for aggravated robbery. ", " He was\r\nconvicted in two separate cases of aggravated robbery, and in another, robbery. ", " Those\r\ncases were consolidated for trial, have been appealed separately, and have been\r\nconsolidated for purposes of briefing.", "\n\tSince the briefs and arguments raised therein are identical in each appeal, for the\r\nreasons stated in Carlton v. State, No. ", "06-02-00071-CR, we likewise find we lack\r\njurisdiction to consider the merits of Carlton's point of error.", "\n\tThe appeal is dismissed for want of jurisdiction.", "\n\n\n\t\t\t\t\t\t\tDonald R. Ross\n\t\t\t\t\t\t\tJustice\n\nDate Submitted:\tOctober 9, 2002\nDate Decided:\tOctober 10, 2002\n\nDo Not Publish\n\r\nssue before us is whether Franklin timely\r\nfiled his notice of appeal. ", " We conclude he did not and dismiss the attempted appeal for want of\r\njurisdiction.", "\n\tOn the issue of whether Franklin timely perfected his appeal, the record establishes\r\n(1) Franklin's sentence was imposed on March 31, 2003; (2) Franklin did not file a motion for new\r\ntrial; and (3) Franklin's notice of appeal was not filed until August 6, 2003.", "\n\tA timely notice of appeal is necessary to invoke this Court's jurisdiction. ", " Olivo v. State, 918\r\nS.W.2d 519, 522 (Tex. ", "Crim. ", "App. ", "1996). ", " Rule 26.2(a) prescribes the time period in which a notice\r\nof appeal must be filed by the defendant in order to perfect an appeal in a criminal case. ", " A\r\ndefendant's notice of appeal is timely if filed within thirty days after the day sentence is imposed or\r\nsuspended in open court, or within ninety days after sentencing if the defendant timely files a motion\r\nfor new trial. ", " Tex. ", "R. App. ", "P. 26.2(a); Olivo, 918 S.W.2d at 522. ", "\n\tWhen a defendant appeals from a conviction in a criminal case, the time to file a notice of\r\nappeal runs from the date sentence is imposed or suspended in open court, not from the date sentence\r\nis signed and entered by the trial court. ", " Rodarte v. State, 860 S.W.2d 108, 109 (Tex. ", "Crim. ", "App.", "\r\n1993). ", " The last date allowed for Franklin to timely file his notice of appeal, without a timely filed\r\nmotion for new trial, was April 30, 2003, thirty days after the day the sentence was imposed. ", "Tex.", "\r\nR. App. ", "P. 26.2(a)(1). ", " Because Franklin did not file his notice of appeal until August 6, 2003, he\r\nfailed to perfect this appeal. ", " Accordingly, we dismiss the appeal for want of jurisdiction.", "\n\n\n\n\t\t\t\t\t\tJack Carter\n\t\t\t\t\t\tJustice\n\nDate Submitted:\tAugust 26, 2003\nDate Decided:\t\tAugust 27, 2003\n\nDo Not Publish\n" ]

{ "pile_set_name": "FreeLaw" }

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[ "package com.github.ltsopensource.tasktracker.cmd;\n\nimport com.github.ltsopensource.cmd.", "HttpCmdProc;\nimport com.github.ltsopensource.cmd.", "HttpCmdRequest;\nimport com.github.ltsopensource.cmd.", "HttpCmdResponse;\nimport com.github.ltsopensource.core.cmd.", "HttpCmdNames;\nimport com.github.ltsopensource.core.commons.utils.", "StringUtils;\nimport com.github.ltsopensource.tasktracker.domain.", "TaskTrackerAppContext;\n\n/**\n * 用于中断某个Job\n * @author Robert HG (254963746@qq.com) on 3/13/16.", "\n */\npublic class JobTerminateCmd implements HttpCmdProc {\n\n private TaskTrackerAppContext appContext;\n\n public JobTerminateCmd(TaskTrackerAppContext appContext) {\n this.appContext = appContext;\n }\n\n @Override\n public String nodeIdentity() {\n return appContext.getConfig().getIdentity();\n }\n\n @Override\n public String getCommand() {\n return HttpCmdNames.", "HTTP_CMD_JOB_TERMINATE;\n }\n\n @Override\n public HttpCmdResponse execute(HttpCmdRequest request) throws Exception {\n\n String jobId = request.getParam(\"jobId\");\n if (StringUtils.isEmpty(jobId)) {\n return HttpCmdResponse.newResponse(false, \"jobId can't be empty\");\n }\n\n if (!", "appContext.getRunnerPool().getRunningJobManager().running(jobId)) {\n return HttpCmdResponse.newResponse(false, \"jobId dose not running in this TaskTracker now\");\n }\n\n appContext.getRunnerPool().getRunningJobManager().terminateJob(jobId);\n\n return HttpCmdResponse.newResponse(true, \"Execute terminate Command success\");\n }\n}\n" ]

{ "pile_set_name": "Github" }

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[ "Q:\n\nHow to set a View Visible on a specific time?", "\n\nI'm trying to make a Video Player android application in which I have to display an ImageButton on the top of Video on a specific Time.", "\nThis is my layout \n<FrameLayout\n android:id=\"@+id/videoSurfaceContainer\"\n android:layout_width=\"match_parent\"\n android:layout_height=\"wrap_content\" >\n\n <SurfaceView\n android:id=\"@+id/videoSurface\"\n android:layout_width=\"wrap_content\"\n android:layout_height=\"wrap_content\" />\n\n <LinearLayout\n android:id=\"@+id/thumbnail\"\n android:layout_width=\"wrap_content\"\n android:layout_height=\"wrap_content\"\n android:layout_gravity=\"center_vertical|end\"\n android:orientation=\"vertical\"\n android:visibility=\"gone\" >\n\n <ImageButton\n android:id=\"@+id/imageButton\"\n android:layout_width=\"wrap_content\"\n android:layout_height=\"wrap_content\"\n android:src=\"@drawable/image\" />\n\n <TextView\n android:id=\"@+id/time\"\n android:layout_width=\"wrap_content\"\n android:layout_height=\"wrap_content\"\n android:text=\"TEST TEST\"\n android:textColor=\"#FFFFFF\" />\n </LinearLayout>\n </FrameLayout>\n\nand i have an ArrayList of type ThumbnailInfo where \npublic class ThumbnailInfo {\n int time; //time in milliseconds\n String body;\n\n ThumbnailInfo(int time,String body){\n this.time = time;\n this.body = body;\n }\n}\n\nSo now I have time stored in the arraylist and i want to display the thumbnail 10 seconds before the specific time.", "\nFor e.g. \nSuppose time stored in arraylist is 00:40 , 01:30 , 04:50 (just to make the question clear ,time is stored in milliseconds).", "\nSo when i'm playing the Video , I have to set the Visibility of thumbnail layout as VISIBLE at 00:30 , 01:20 , 04:40 and will set the visibility GONE at 00:41, 01:31 , 04:51\nSo my question is How can I check stored Time in ArrayList continuously and execute the above mentioned operation.", "\nCurrently what i have is the Current Position of the Video using mediaPlayer.getCurrentPosition();\nNow I have to compare the current position of the video continuously with the time stored in ArrayList . ", "\nIs there any observer which can make the task easy or any other approach.", "\nP.S. : User can pause the video multiple times.", "\nAny help is welcome. ", "Thanx in advance !!!", "\n\nA:\n\nTo simple you should convert time from minute to second start with 0. ", "Such as:\n\nVISIBLE at 00:30 , 01:20 , 04:40 => ArrayList< Visible > with values 30, 80, 280\nINVISIBLE at 00:41, 01:31 , 04:51 => ArrayList< Invisible > with values 41, 61, 281\n int seconds = 0;\n CountDownTimer t = new CountDownTimer(Long.", "MAX_VALUE ,1000) { // interval 1s \n\n // This is called every interval. (", "Every 1 seconds in this example)\n public void onTick(long millisUntilFinished) {\n checkVisible(ArrayList visible);// make visible button\n checkInvisible(ArrayList invisible); // make invisible button\n seconds++:\n }\n public void onFinish() {\n System.out.println(\"finished\"); \n seconds = 0; \n }\n }.start();\n\nRemember call \n\nt.cancel() when pause video\nt.finish() when finish play video\nreset seconds variable;\n\n" ]

{ "pile_set_name": "StackExchange" }

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[ "INTRODUCTION {#rrw097s1}\n============\n\nExposure to ionizing radiation is an established risk factor for the development of breast cancer. ", "Epidemiological studies of the Japanese atomic bomb survivors indicate that the breast is one of the most susceptible organs to radiation-induced carcinogenesis \\[[@rrw097C1], [@rrw097C2]\\]. ", "Other studies on patients exposed to medical radiation provide evidence that there is a high risk of second breast cancer among women treated with chest radiotherapy for Hodgkin lymphoma or benign breast disease \\[[@rrw097C3], [@rrw097C4]\\]. ", "However, the molecular mechanisms underlying radiation-induced human breast cancer are yet to be fully characterized, because breast cancer in humans occurs as a result of the combined effects of radiation and other factors such as age, diet, family history, alcohol consumption, and hormone therapy \\[[@rrw097C5]\\].", "\n\nAnimal models are useful for studying the contribution of a single factor in carcinogenesis, such as ionizing radiation and chemical carcinogens. ", "The rat model of mammary tumor is widely used to study breast cancer risk and mechanisms of breast carcinogenesis, because rat mammary tumors are comparable in many aspects to human breast tumors, such as in their high frequency of hormone dependence and pathological processes of malignant transformation; i.e. progression from ductal hyperplasia to ductal carcinoma *in situ* (DCIS) and carcinoma \\[[@rrw097C6]\\]. ", "Using rat models, mammary tumors induced by ionizing radiation and chemical carcinogens have been shown to exhibit specific genomic aberrations \\[[@rrw097C7]\\]. ", "It has been repeatedly reported that *Hras* mutations are frequently observed in rat mammary carcinomas induced by exposure to 1-methyl-1-nitrosourea (MNU), but not in radiation-induced carcinomas \\[[@rrw097C8]--[@rrw097C11]\\]. ", "Loss of heterozygosity (LOH) of several chromosomal regions has been found in 2-amino-1-methyl-6-phenylimidazo\\[4,5-b\\]pyridine (PhIP)-induced rat mammary carcinomas \\[[@rrw097C12], [@rrw097C13]\\], whereas LOH is a rare event in carcinomas induced by ionizing radiation and MNU \\[[@rrw097C14], [@rrw097C15]\\]. ", "A study of radiation-induced mammary carcinomas in Sprague--Dawley (SD) rats failed to identify any recurrent loss of DNA copy number \\[[@rrw097C16]\\]. ", "While female SD rats are known to be highly susceptible to spontaneous or radiation-induced mammary carcinomas \\[[@rrw097C17]\\], the F1 hybrids of SD rats and mammary cancer--resistant Copenhagen (COP) rats have a very low incidence of spontaneous mammary carcinomas, yet still have a high incidence of radiation-induced carcinomas. ", "Our earliest investigations with carcinomas from this F1 strain suggest that genes of the phosphatidylinositol 3-kinase (PI3K)/AKT pathway might be potential targets of genomic deletion and/or allele loss (unpublished data).", "\n\nThe PI3K/AKT pathway regulates many cellular functions, such as cell proliferation, growth, survival and motility \\[[@rrw097C18], [@rrw097C19]\\] and is frequently mutated or altered in human cancers \\[[@rrw097C20], including a majority of breast cancers \\[[@rrw097C21]--[@rrw097C23]\\]. ", "Recently, it has been reported that ionizing radiation induces persistent changes in the expression of genes involved in the PI3K/Akt pathway and causes activation of the pathway in normal rodent mammary glands \\[[@rrw097C24]--[@rrw097C26]\\], suggesting a link between this pathway and radiation-induced mammary carcinogenesis. ", "Furthermore, our previous investigation revealed that mTOR activity, a downstream target of the PI3K/Akt pathway, was higher in some radiation-induced rat mammary carcinomas than in MNU-induced carcinomas \\[[@rrw097C27]\\].", "\n\nIn the present study, we performed immunohistochemical staining of phospho-AKT1 to assess pathway activation in SD × COP F1 rat mammary carcinomas induced by radiation, with MNU-induced carcinomas analyzed in parallel for comparison. ", "Further, gene expression and sequence analyses for several candidate genes in the PI3K/Akt pathway were examined to investigate the mechanisms underlying pathway activation. ", "Our results suggest that although AKT phosphorylation is a common feature in a subset of cells within rat mammary carcinomas induced by either agent, there are no mutations in key pathway genes or permanent changes to gene expression that correlate with the level of activation in individual carcinomas. ", "Although it is possible that a number of subtle changes might collectively induce AKT phosphorylation, or that activation of the pathway may be important at an earlier stage of carcinoma development, it seems unlikely that radiation-induced alterations of the PI3K/AKT pathway genes are critical events in radiation-carcinogenesis of the rat mammary gland.", "\n\nMATERIALS AND METHODS {#rrw097s2}\n=====================\n\nTumor samples {#rrw097s2a}\n-------------\n\nAnimal experiments were approved by the Institutional Animal Care and Use Committee of the National Institute of Radiological Sciences (NIRS). ", "In this study, F1 hybrid rats were created by crossing mammary cancer--susceptible and --resistant rats, Sprague--Dawley (SD) and Copenhagen (COP) rats , respectively. ", "Female SD (Jcl:SD) rats were purchased from CLEA Japan, Inc. (Tokyo, Japan). ", "Male and female COP (COP/Hsd and Iar:COP) rats were obtained from Harlan Sprague Dawley, Inc. (Madison, WI, USA) and the Institute for Animal Reproduction (Ibaraki, Japan), respectively, and were maintained by brother--sister mating. ", "The breeding of the (SD × COP) F1 hybrid rats was performed in our facility at the NIRS. ", "They were maintained under specific pathogen-free (SPF) conditions with controlled temperature (23°C ± 1°C) and humidity (45% ± 5%) under a regular 12 h light--dark cycle. ", "They were fed a standard CE-2 diet (CLEA, Japan) and sterile water *ad libitum*. ", "At 7 weeks of age, female F1 rats (*n* = 29) were exposed to whole-body ɣ-radiation (4 Gy) using a ^137^Cs source (Gammacell; Nordion International, Ottawa, Canada) at a dose rate of 0.5 Gy/min. ", "Female F1 rats (*n* = 30) received a single intraperitoneal injection of 50 mg MNU/kg body weight at 7 weeks. ", "The treated rats were observed daily, and weekly palpation was started at 2 weeks after the treatments to identify mammary tumors. ", "The animals that showed general deterioration during the observation period (including body weight loss and severe anemia due to tumor development) were euthanized; any mammary cancers were collected, including those discovered post-mortem in rats that were found dead. ", "The characteristics of the carcinomas are shown in Tables [1](#rrw097TB1){ref-type=\"table\"} and [2](#rrw097TB2){ref-type=\"table\"}. ", "The majority of radiation- and MNU-induced carcinomas (12 of 14 and 10 of 10, respectively) were of luminal subtype. ", "Mammary carcinomas and normal glands from carcinoma-bearing rats were frozen and stored at −80°C. ", "Table 1.Rat mammary carcinomas induced by ionizing radiationCase No.", "Tumor weight (g)Latency period (weeks)Immunohistochemical scoreSubtypeGene expression fold change^[a](#rrw097tfn2){ref-type=\"fn\"}^Percentage of p-Akt immunopositive carcinoma cellsERPRHER2Ki-67OncogenesTumor suppressor genes*Erbb2Pik3caPdk1AktPtenPik3r1Map3k1Inpp4b*J-23193321Luminal A1.70.910.550.730.390.470.320.331J-49.9113311Luminal An.d.n.d.n.d.n.d.n.d.n.d.n.d.n.d.51J-6^[b](#rrw097tfn3){ref-type=\"fn\"}^32152013Luminal B0.681.50.650.520.581.40.480.2944J-136.2553221Luminal A2.51.40.870.651.10.450.690.714J-142.9513223Luminal B1.91.50.880.700.700.390.450.2130J-16^[c](#rrw097tfn4){ref-type=\"fn\"}^8.4343212Luminal B1.20.530.390.330.350.150.400.1333J-180.2669^[d](#rrw097tfn5){ref-type=\"fn\"}^3321Luminal An.d.n.d.n.d.n.d.n.d.n.d.n.d.n.d.5J-200.35503221Luminal A0.300.240.800.790.590.871.30.1941J-262.2713322Luminal B1.51.61.01.11.11.31.60.4524J-270.84543213Luminal B0.724.42.21.31.23.01.80.8269J-321.5833321Luminal A3.22.71.00.630.361.62.20.3355J-360.98610011Triple negativen.d.n.d.n.d.n.d.n.d.n.d.n.d.n.d.23J-3910850011Triple negative1.81.31.71.70.660.340.421.778J-400.5097^[d](#rrw097tfn5){ref-type=\"fn\"}^3322Luminal Bn.d.n.d.n.d.n.d.n.d.n.d.n.d.n.d.33Average7.6532.52.01.61.6n.a.1.51.61.00.840.700.991.00.5235(SD)(11)(28)(1.1)(1.2)(0.51)(0.84)(0.88)(1.2)(0.56)(0.40)(0.32)(0.86)(0.85)(0.47)(23)[^1][^2][^3][^4][^5]Table 2.Rat mammary carcinomas induced by MNUCase No.", "Tumor weight (g)Latency period (weeks)Immunohistochemical scoreSubtypeGene expression fold change^[a](#rrw097tfn7){ref-type=\"fn\"}^Percentage of p-Akt immunopositive carcinoma cellsERPRHER2Ki-67OncogenesTumor suppressor genes*Erbb2Pik3caPdk1AktPtenPik3r1Map3k1Inpp4b*K2811 Mt-122453321Luminal A2.072.650.941.150.860.580.581.065.2K2896 Mt-13.8523332Luminal B3.100.920.571.290.510.350.450.239.9K3019 Mt-165433332Luminal B2.631.250.440.880.510.201.200.120.5K3052 Mt-10.65533321Luminal A2.781.160.591.051.920.321.260.0629K3064 Mt-18.1713321Luminal A6.373.701.041.601.031.243.740.3729K3113 Mt-10.88743333Luminal B2.351.830.972.590.850.924.960.5120K3154 Mt-1^[b](#rrw097tfn8){ref-type=\"fn\"}^18540321Luminal A1.550.840.380.690.340.271.320.163.3K3154 Mt-22.3543321Luminal A6.722.340.871.120.760.831.810.4218K3158 Mt-1^[c](#rrw097tfn9){ref-type=\"fn\"}^2.0543331Luminal A3.851.300.570.980.630.311.780.379.7K3160 Mt-10.76623331Luminal A19.142.610.831.560.672.722.080.7656Average12562.73.02.51.4n.a.5.11.90.721.30.810.771.90.4118(SD)(20)(10)(0.95)(0.0)(0.53)(0.69)(5.2)(0.94)(0.24)(0.54)(0.44)(0.76)(1.4)(0.31)(17)[^6][^7][^8][^9]\n\nDNA and RNA sample preparation {#rrw097s2b}\n------------------------------\n\nGenomic DNA and total RNA were both extracted from the same normal mammary gland and mammary carcinoma frozen samples using the AllPrep DNA/RNA Mini Kit (Qiagen, Tokyo, Japan).", "\n\nQuantitative RT-PCR {#rrw097s2c}\n-------------------\n\nComplementary DNA (cDNA) was synthesized by reverse transcription (ReverTra Ace Toyobo, Osaka, Japan). ", "One sample from an animal found dead and two samples with low RNA yield among the radiation-induced carcinoma samples were not subjected to quantitative expression analysis. ", "Primers were designed for eight genes (*Erbb2/HER2, Pik3ca, Pdk1, Akt1, Pten, Pik3r1*, *Map3k1* and *Inpp4b*), using the Primer Express 3.0 software (Applied Biosystems), and were validated to amplify a single product of the correct size for each gene by agarose gel electrophoresis. ", "The primer sequences and PCR conditions are provided in [Supplementary Table S1](#sup1){ref-type=\"supplementary-material\"}. ", "PCR was performed using the Stratagene Mx3000P real-time PCR system (Agilent Technologies, Santa Clara, CA, USA) and SYBR Premix Ex Taq (Takara Bio Inc., Otsu, Japan). ", "The expression level of the target genes was normalized to that of *Actb*. ", "Expression data from one matched normal mammary gland among the radiation-induced carcinoma samples was excluded from further analysis due to poor amplifiability of *Actb*, which was more than 100-fold lower than the median value.", "\n\nSequence analysis {#rrw097s2d}\n-----------------\n\nMutations in all exons of *Pten*, *Pik3ca, Pik3r1* and *Map3k1* genes were screened initially by direct sequencing of PCR products from cDNAs from the twelve radiation- and ten MNU-induced carcinoma samples with both DNA and RNA available, and identified mutations were confirmed by direct sequencing of PCR products amplified from genomic DNA on an Applied Biosystems 3130 Genetic Analyzer using a BigDye Terminator kit (Life Technologies Inc., Carlsbad, CA, USA). ", "The primer sequences and PCR conditions are provided in [Supplementary Table S2](#sup1){ref-type=\"supplementary-material\"}. ", "All sequences were obtained in both directions. ", "Sequence analysis was done using the ATGC and GENETYX software (Genetyx Corporation, Tokyo, Japan). ", "An in-frame deletion in *Pik3r1* in one carcinoma was confirmed by TA cloning of PCR products amplified from genomic DNA, and subsequent sequencing (TOPO TA cloning kit, Invitrogen, Life Technologies, Tokyo, Japan).", "\n\nImmunohistochemical staining {#rrw097s2e}\n----------------------------\n\nFormalin-fixed, paraffin-embedded tissue sections (\\~3 µm thick) were immunostained with primary antibodies against estrogen receptor α (ER) (6F11, Leica Microsystems, Tokyo, Japan); progesterone receptor (PR) (PR-2C5, Invitrogen, Carlsbad, CA); human epidermal growth factor receptor 2 (HER2) (A0485, DAKO Japan, Tokyo, Japan); Ki-67 (M7248, DAKO Japan, Tokyo, Japan); or phospho-T308-Akt1 (ab66134, Abcam, Cambridge, UK), as described previously \\[[@rrw097C28]\\]. ", "Briefly, sections were deparaffinized and rehydrated, and antigen retrieval was performed with autoclave treatment at 120°C in 10 mM sodium citrate buffer (pH 6.0) for 20 min for ER, PR, HER2 and Ki67 and with microwave treatment at 98°C for 20 min for phospho-T308-Akt1. ", "After blocking endogenous peroxidase and non-specific binding sites, specimens were incubated at 4°C overnight for ER, PR, HER2 and phospho-T308-Akt1 or at room temperature overnight for Ki-67. ", "Then, sections were treated with a peroxidase-conjugated secondary antibody (Histofine Simple Stain MAX-PO, Nichirei, Tokyo, Japan). ", "Immunoreaction was visualized by using the 3, 3'-diaminobenzidine (DAB) Peroxidase Substrate Kit (Vector Laboratories, Burlingame, CA, USA). ", "Sections were counterstained with hematoxylin. ", "The intensity of staining was graded from 0 to 3 for ER and PR (0 = \\<1% of all cells with nuclear staining; 1 = 1--9%; 2 = 10--20%; 3 = \\>20%) and for Ki-67 (0 = \\<5% of all cells with nuclear staining; 1 = 5--9%; 2 = 10--20%; 3 = \\>20%), according to published studies \\[[@rrw097C29], [@rrw097C30]\\]. ", "The intensity of membrane expression was graded from 0 to 3 for HER2 according to the DAKO-score. ", "Cancer subtype was also determined according to the St Gallen consensus \\[[@rrw097C31]\\]. ", "To calculate the percentage of phospho-AKT1--positive carcinoma cells, stained sections were scanned using a NanoZoomer 2.0-HT slide scanner (Hamamatsu Photonics, Hamamatsu, Japan) and analyzed with the Definiens Tissue Studio 3.0 software (Definiens, Munich, Germany). ", "A carcinoma sample containing \\>10% of phospho-AKT1--positive carcinoma cells in the tissue was considered as positive for phospho-AKT1 \\[[@rrw097C32]\\].", "\n\nStatistical analysis {#rrw097s2f}\n--------------------\n\nStatistical analysis was carried out with the StatMate software version 3.0 (ATMS, Tokyo, Japan). ", "Comparison between two groups was done using a non-parametric Mann--Whitney *U*-test. *", "P* \\< 0.05 was considered to be statistically significant.", "\n\nRESULTS {#rrw097s3}\n=======\n\nActivation of the PI3K/Akt signaling pathway {#rrw097s3a}\n--------------------------------------------\n\nFirst, we assessed activation of the PI3K/Akt pathway in the mammary carcinomas (Figs [1](#rrw097F1){ref-type=\"fig\"} and [2](#rrw097F2){ref-type=\"fig\"}), where we observed that 79% (11 out of 14 cases) of radiation-induced carcinomas and 50% (5 out of 10 cases) of MNU-induced carcinomas were positive for phospho-AKT1 (Tables [1](#rrw097TB1){ref-type=\"table\"} and [2](#rrw097TB2){ref-type=\"table\"}). ", "Some of the carcinomas showed heterogeneous staining patterns for phospho-AKT1, indicating heterogeneous carcinoma cell populations (Figs [1](#rrw097F1){ref-type=\"fig\"}E and F and [2](#rrw097F2){ref-type=\"fig\"}E and F). ", "In order to determine the cause of the pathway\\'s activation, we measured the gene expression level for key genes in the PI3K/Akt signaling pathway, comparing against age-matched normal mammary gland tissues. ", "Looking at the negative pathway regulators, the expression of *Pten* was significantly decreased in radiation-induced carcinomas (Fig. [", "3](#rrw097F3){ref-type=\"fig\"}), while the decrease was not significant for MNU-induced carcinomas; *Pik3r1* levels were significantly lower only in MNU-induced carcinomas (Fig. [", "4](#rrw097F4){ref-type=\"fig\"}), and *Inpp4b* levels were decreased in carcinomas of both groups. ", "For the positive regulators, *Erbb2* and *Pik3ca* were both significantly upregulated in MNU-induced carcinomas, while interestingly, *Pdk1* was significantly decreased in MNU-induced carcinomas. ", "The phospho-AKT1 staining level was positively correlated with *Pdk1* gene expression, but only in the radiation-induced carcinomas (Fig. [", "5](#rrw097F5){ref-type=\"fig\"} and [Supplementary Figure S1](#sup1){ref-type=\"supplementary-material\"}), while positive correlations with *Erbb2* and *Pik3r1 g*ene expression levels were observed for MNU-induced carcinomas (Fig. [", "5](#rrw097F5){ref-type=\"fig\"} and [Supplementary Figure S2](#sup1){ref-type=\"supplementary-material\"}). ", "The weak or absent correlations of the expression of the various regulators with phosphorylation of AKT (as a readout of pathway activation) may be an indication of changes at the level of protein function, or subtle changes across a number of pathway inputs that can collectively alter the state of the network, rather than perturbation of a single signaling node.", "\n\n{#rrw097F1}\n\n{#rrw097F2}\n\n{#rrw097F3}\n\n{#rrw097F4}\n\n{#rrw097F5}\n\nMutations in the PI3K/Akt signaling pathway {#rrw097s3b}\n-------------------------------------------\n\nTo assess whether any of the genes in the pathway had been activated/inactivated by acquired mutations, we sequenced *Pten*, *Pik3ca*, *Pik3r1* and *Map3k1*, each of which are documented to be mutated in human mammary cancers, plus the *Hras* gene which is known to be mutated at high frequency in chemically induced rat mammary carcinomas \\[[@rrw097C10]\\]. ", "Although *Hras* was mutated in 8 out of the 10 MNU-induced carcinomas (all at codon 12), interestingly, there were no *Hras* mutations in the radiation-induced carcinomas. ", "The same heterozygous missense mutation in *Pik3ca* was identified in one radiation-induced carcinoma (Fig. [", "6](#rrw097F6){ref-type=\"fig\"}) and one MNU-induced carcinoma (Fig. [", "7](#rrw097F7){ref-type=\"fig\"}); it is predicted to disrupt the helical domain, with an additional MNU-induced carcinoma harboring two independent missense mutations (in the C2 and kinase domains). ", "An in-frame deletion in *Pik3r1* was identified in a single radiation-induced carcinoma (Fig. [", "8](#rrw097F8){ref-type=\"fig\"}), while no mutations were identified in *Pten* or *Map3k1* in any of the carcinomas. ", "These are also summarized in Table [3](#rrw097TB3){ref-type=\"table\"}. ", "Table 3.Mutations in the *Pik3r1* and *Pik3ca* genes of radiation- and MNU-induced rat mammary carcinomasCase No.", "GeneDNAExonmRNAMutaion typeDomain**J-16***Pik3r1*1708_1725del12570_575delin-frame deletioniSH2**J-6***Pik3ca*g1624a9E542KmissenseHelical**K3158_Mt1***Pik3ca*g1624a9E542KmissenseHelical**K3154_Mt1***Pik3ca*c1215t6S405FmissenseC2*Pik3ca*a2816g19D939GmissenseKinase\n\n{#rrw097F6}\n\n{#rrw097F7}\n\n{#rrw097F8}\n\nDISCUSSION {#rrw097s4}\n==========\n\nThe products of the *Pten*, *Pik3r1* and *Inpp4b* genes function as negative regulators of PI3K activity, either by controlling the levels of PIP3, which mediates Akt phosphorylation, or by directly increasing PI3K activity \\[[@rrw097C33]\\]. ", "Consistent with our results, it has been reported that loss-of-function mutations of *Pten* in human breast cancer are much less common than promoter methylation and reduced expression levels \\[[@rrw097C34]--[@rrw097C37]\\], with reduced expression of *Inpp4b* also reported in human breast cancers \\[[@rrw097C38], [@rrw097C39]\\]. ", "However, no direct correlation was observed between the staining levels of phospho-AKT1 and gene expression levels of *Pten* and *Inpp4b* in radiation- and MNU-induced rat mammary carcinomas, suggesting that while their downregulation may permit AKT activation, it does not explain the activation of the pathway. ", "Consistent with a previous report \\[[@rrw097C40]\\], our results suggest that increased *Erbb2* gene expression might contribute to AKT phosphorylation, but this was only evident in the MNU-induced carcinomas, while a contribution from *Pdk1* expression was only observed in radiation-induced carcinomas. ", "However, these correlations were both heavily influenced by one or two carcinomas with both high phospho-AKT1 staining and high *Erbb2/Pdk1* gene expression, which might indicate a causal relationship in rare specific carcinomas, but little influence in the remaining majority of carcinomas. ", "Such a mechanism is consistent with the counter-intuitive positive correlation of *Pik3r1* with phospho-AKT1 staining, where dysregulation of the signaling network in a single carcinoma may result in anomalous expression of a negative regulator while still resulting in pathway activation. ", "Where there are a large number of positive and negative signaling inputs, it is possible that no single protein will show a strong direct influence on the activation of the pathway, since subtle changes of both positive and negative regulators may collectively influence the phosphorylation of AKT, and/or individual carcinomas may utilize different routes to pathway activation \\[[@rrw097C41]\\] that are thus not generalizable across a group of carcinomas, making it difficult to identify statistically significant interactions.", "\n\nThe in-frame deletion detected in *Pik3r1* is predicted to disrupt the amino-acid residues from 572 to 600 in the iSH2 domain of the p85α regulatory subunit of PI3K \\[[@rrw097C42]\\], which inhibits PI3K activity, a mutation also found in human breast cancers \\[[@rrw097C43], [@rrw097C44]\\], though overall, mutation of *Pik3r1* is rare \\[[@rrw097C45], [@rrw097C46]\\]. ", "The E542K mutation in the *Pik3ca* gene observed in the radiation- and MNU-induced mammary carcinomas is a prominent hotspot mutation leading to constitutive activation of AKT in human breast cancer \\[[@rrw097C47]\\], with *Pik3ca* mutations more prevalent in human breast cancer overall \\[[@rrw097C45], [@rrw097C48]\\] despite being rare in these groups of rat mammary carcinomas. ", "Yet, the small number of carcinomas with mutations in these PI3K/AKT pathway genes did not show any more pathway activation than the carcinomas without mutations, in some cases quite the opposite, casting doubt on the importance of these acquired mutations. ", "One explanation is that activation of the PI3K/AKT pathway may facilitate a particular step in mammary carcinoma development or growth, and that once this barrier is passed, ongoing signaling through this pathway may become redundant. ", "This would have the effect of disconnecting the final phospho-AKT1 levels in the late-stage carcinoma from the state of the network at the critical stage. ", "In many of the carcinomas, the phospho-AKT1 staining was highly heterogeneous, which may also interfere with the correlation of the various signaling proteins in DNA/RNA from a mixed tissue sample. ", "Comparison against gene expression levels in normal mammary tissue that was not exposed to either radiation or a chemical carcinogen might also have revealed potential gene expression changes induced by these agents in non-cancer tissues.", "\n\nTogether, these data suggest that while AKT phosphorylation was a common feature in a subset of cells within rat mammary carcinomas induced by radiation or a chemical carcinogen, this was not usually mediated by either disrupted function by mutation of relevant signaling proteins nor correlated with the final expression levels of these same key genes at the advanced cancer stage, although general increases/decreases could be observed when compared with normal mammary tissue. ", "Our findings suggest that while AKT phosphorylation within carcinoma cells may contribute to cancer growth in particular cases, it is not a major underlying causal event mediated by permanent alteration of the signaling pathway genes; nor does it appear to be more likely to be involved in mammary carcinomas induced by exposure to radiation compared with induction by a canonical chemical carcinogen. ", "Understanding the mechanisms that underlie the induction of breast cancer following exposure to radiation is essential for future efforts to minimize risk and develop effective countermeasures.", "\n\nSUPPLEMENTARY DATA {#rrw097s5}\n==================\n\n[Supplementary data are available at the *Journal of Radiation Research* online](#sup1){ref-type=\"supplementary-material\"}.", "\n\nACKNOWLEDGEMENTS {#rrw097s6}\n================\n\nThe authors thank M. Ootawara, H. Osada and I. Tanaka for technical assistance, and the Department of Engineering and Safety, QST NIRS, for animal management. ", "The results of this study were presented at the 57th Annual Meeting of the Japan Radiation Research Society.", "\n\nCONFLICT OF INTEREST {#rrw097s7}\n====================\n\nThe authors declare that there are no conflicts of interest.", "\n\nFUNDING {#rrw097s8}\n=======\n\nThis work was supported in part by a Grant-in-Aid for the Third Comprehensive 10-Year Strategy for Cancer Control and for Cancer Research from the Ministry of Health, Labor and Welfare of Japan. ", "The animals used in this experiment are part of the J-SHARE project (Japan-StoreHouse of Animal Radiobiology Experiments) under construction by the QST National Institute of Radiological Sciences.", "\n\nSupplementary Material\n======================\n\n###### \n\nClick here for additional data file.", "\n\n###### \n\nClick here for additional data file.", "\n\n###### \n\nClick here for additional data file.", "\n\n###### \n\nClick here for additional data file.", "\n\n[^1]: ER = estrogen receptor α, PR = progesterone receptor, HER2 = human epidermal growth factor receptor 2, n.d. ", "= no data, n.a. ", "= not applicable.", "\n\n[^2]: ^a^Data denote *Actb*-normalized fold changes relative to matched normal mammary glands.", "\n\n[^3]: ^b^Sample with a hotspot mutation in exon 9 (codon E542K) of *Pik3ca* gene.", "\n\n[^4]: ^c^Sample with an 18-bp deletion in exon 12 (codon 570--575del) of *Pik3r1* gene.", "\n\n[^5]: ^d^Sample that had been nonpalpable until dissection.", "\n\n[^6]: ER = estrogen receptor α, PR = progesterone receptor, HER2 = human epidermal growth factor receptor 2, n.a. ", "= not applicable.", "\n\n[^7]: ^a^Data denote *Actb*-normalized fold changes relative to matched normal mammary glands.", "\n\n[^8]: ^b^Sample with a hotspot mutation in exons 6 and 19 (codons S405F and D939G, respectively) of *Pik3ca* gene.", "\n\n[^9]: ^c^Sample with a hotspot mutation in exon 9 (codon E542K) of *Pik3ca* gene.", "\n" ]

{ "pile_set_name": "PubMed Central" }

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[ "Q:\n\n.htaccess - Convert PHP GET to URL Path but keep GET in URL\n\nMy Goal was to redirect a URL like http://example.com/c/themeforest/wordpress to the PHP http://example.com/?params=c/themeforest/wordpress. ", "\nTo achieve this, I used a .htaccess file with the following contents:\nOptions +FollowSymLinks\nRewriteEngine On\nRewriteCond %{REQUEST_FILENAME} !", "-d\nRewriteCond %{REQUEST_FILENAME} !", "-f\nRewriteRule ^(.*)$ index.php?params=$1 [NC]\n\nBut now I have to also be able to redirect URLs like http://example.com/c/themeforest/wordpress?p=2\nto http://example.com/?params=c/themeforest/wordpress&p=2 for multiple Pages.", "\nThe Page Number has to be sent in a seperate PHP GET Variable.", "\n\nA:\n\nAdd the QSA (query string append) flag, so:\nRewriteRule ^(.*)$ index.php?params=$1 [NC,QSA]\n\n" ]

{ "pile_set_name": "StackExchange" }

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[ " FILED\n NOT FOR PUBLICATION JAN 04 2012\n\n MOLLY C. DWYER, CLERK\n UNITED STATES COURT OF APPEALS U .S. C O U R T OF APPE ALS\n\n\n\n\n FOR THE NINTH CIRCUIT\n\n\n\nUNITED STATES OF AMERICA, Nos. ", "10-10572\n 10-10574\n Plaintiff - Appellee,\n D.C. Nos. ", "2:07-cr-00488-JAT\n v. 2:98-cr-00391-JAT\n\nTRUMAN HARLOW STEVENS, Jr.,\n MEMORANDUM *\n Defendant - Appellant.", "\n\n\n\n Appeals from the United States District Court\n for the District of Arizona\n James A. Teilborg, District Judge, Presiding\n\n Submitted December 19, 2011 **\n\nBefore: GOODWIN, WALLACE, and McKEOWN, Circuit Judges.", "\n\n In these consolidated appeals, Truman Harlow Stevens, Jr., appeals from the\n\nconsecutive 24-month and 36-month sentences imposed upon revocation of two\n\nconcurrent terms of supervised release. ", "We have jurisdiction under 28 U.S.C.\n\n§ 1291, and we affirm.", "\n\n *\n This disposition is not appropriate for publication and is not precedent\nexcept as provided by 9th Cir. ", "R. 36-3.", "\n **\n The panel unanimously concludes this case is suitable for decision\nwithout oral argument. ", "See Fed. ", "R. App. ", "P. 34(a)(2).", "\n\f Stevens contends that his sentences are substantively unreasonable because\n\nthe district court placed undue emphasis on the need to punish him for the\n\nseriousness of his underlying convictions and violations at the expense of other\n\nmore salient factors. ", "The record reflects that the district court considered and\n\nproperly applied the relevant factors before imposing sentences that are not\n\nunreasonable under the totality of the circumstances and in light of the sentencing\n\nfactors set forth in 18 U.S.C. §§ 3553(a) and 3583(e). ", "See United States v. Miqbel,\n\n444 F.3d 1173, 1182 (9th Cir. ", "2006).", "\n\n AFFIRMED.", "\n\n\n\n\n 2 10-10572, 10-10574\n\f" ]

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[ "Previous Next 1 of 3 NASA/JPL-Caltech/SwRI/ASI/INAF/JIRAM NASA/JPL-Caltech/SwRI/ASI/INAF/JIRAM NASA/JPL-Caltech/SwRI/ASI/INAF/JIRAM\n\nJupiter is a big, complex, chaotic planet. ", "It has long been known as the most dominant feature in our sun’s orbit, but it wasn’t until last May that the planet’s internal features began to be revealed. ", "During a few close passes, NASA’s $1.1 billion Juno spacecraft collected data on the gas giant that revealed cyclones the size of Earth and a surprisingly strong magnetic field.", "\n\nNow, data collected by Juno have uncovered more never-before-seen features on its north and south poles. ", "In a study published this week in the journal Nature, a team of scientists report bizarre geometric storms surrounding a single massive cyclone on each of the planet’s poles. ", "The storms measure seven thousands miles across and reach wind speeds nearing 220 miles per hour, which would classify them as Category 5 hurricane here on Earth.", "\n\nOn July 4, 2016, Juno rendezvoused with Jupiter to perform a series of close passes that would bring the spacecraft just a couple thousands miles above the gas giant’s top cloud layer. ", "Using its sophisticated instruments, Juno began to peer beneath the planet’s clouds for the first time, snapping photos and measuring Jupiter’s infrared, microwave, ultraviolet, gravity, and magnetic features.", "\n\nAmong the many mysteries Juno scientists sought to uncover were Jupiter’s poles, previously hidden from our telescopes due to the planet’s nonexistent tilt. ", "When Juno finally beamed back images of the north pole, scientists were shocked to find eight cyclones circling a single storm in the middle. ", "Later, the south pole presented a similar arrangement with five outer storms.", "\n\nAlberto Adriani, a co-investigator for Juno’s Jovian Infrared Auroral Mapper (JIRAM) and lead author of the paper, explained that the storms are likely the result of Jupiter’s heat and rotation speed.", "\n\n“The high rotation of the planet — about 10 hours for a complete turn around its axis — and the heat coming from the lower levels of the atmosphere certainly have a great impact to the formation of the cyclonic pattern we have observed over the Jupiter’s poles,” he told Digital Trends.", "\n\nAs with many space discoveries, it’s not always clear how or why the findings are relevant to us on Earth. ", "Sure, Jupiter’s strange storms are cool, but why does it matter?", "\n\n“Space research has a triple value,” Adriani said. ", "There’s the knowledge itself, the search for which “pushes our minds to try to understand what we don’t know.” ", "Then there are the technological advances that enable such a discovery in the first place, some of which can be used to study things like space weather, which have an immediate impact on Earth.", "\n\n“Last but not least, economically speaking every euro [or dollar] invested in research comes back to the society,” Adriani, and that return on investment is often multiplied by many factors.", "\n\nEditors' Recommendations" ]

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[ "# isort:skip_file\nfrom __future__ import absolute_import, division, print_function, unicode_literals\n\nimport torch_glow\nimport torch\n\nfrom tests.utils import GLOW_NODE_NAME, SUBGRAPH_ATTR\nimport unittest\n\n\nclass TestMinGraphSize(unittest.", "TestCase):\n def test_min_graph_size(self):\n \"\"\"Test Glow fuser minimum fusion group size mechanism.\"\"\"", "\n\n def f(a, b, c):\n return (a * a * a * a) / (b * b * b) / (c * c * c * c * c)\n\n torch_glow.disableFusionPass()\n\n # Disable aten::div so that each group of aten::mul nodes will be forced\n # into separate subgraphs\n torch_glow.setFusionBlacklist([\"aten::div\"])\n\n # Set minimum fusion group size to 3 nodes so that the smallest group which\n # contains only 2 nodes will not be created\n torch_glow.setMinFusionGroupSize(3)\n\n a = torch.randn(5, 5)\n b = torch.randn(5, 5)\n c = torch.randn(5, 5)\n\n jit_f = torch.jit.trace(f, (a, b, c))\n jit_f_graph = jit_f.graph_for(a, b, c)\n\n # print(\"before: \", jit_f_graph)\n\n torch_glow.glowCustomFuseDebug_(jit_f_graph)\n\n # print(\"after: \", jit_f_graph)\n\n fusion_nodes = 0\n for node in jit_f_graph.nodes():\n if node.kind() == GLOW_NODE_NAME:\n fusion_nodes += 1\n\n assert fusion_nodes == 2, \"Expected smallest fusion group to not be created\"\n\n torch_glow.clearFusionBlacklist()\n torch_glow.setMinFusionGroupSize(0)\n" ]

{ "pile_set_name": "Github" }

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[ "Thursday, 10 February 2011\n\nBig Show Interview Highlights\n\nBig Show is one World Wrestling Entertainment superstar who couldn’t be happier to have WrestleMania 28 in Miami next year.", "\n\nThe Miami (Pinecrest) resident, real name Paul Wight, looks forward to having the biggest spectacle in sports entertainment in his backyard.", "\n\n“I live here, and I have become part of the community,” said the 7-0, 500-pound superstar. “", "To know how much WreslteMania weekend is going to impact the local community with the influx of money going into the economy with hotels, transportation services, food and all the people who will come here to experience this rich culture and everything WrestleMania has to offer is amazing. ", "My main thing is it’s going to be a short drive to work for me.”", "\n\nBig Show made a surprise appearance at the press conference for the big “Mania in Miami” announcement on Wednesday afternoon at the Fontainebleau Miami Beach.", "\n\nThe 305 brings something unique to the table for WrestleMania, WWE and its universe.", "\n\n“The flavor of Miami, with the salsa, the music, the food,” Show said. “", "You can’t even compare Miami to anywhere else. ", "Miami is not New York. ", "Miami is not Los Angeles. ", "Miami is Miami.”", "\n\nThe big event will emanate from the Miami Dolphins house at Sun Life Stadium on April 1, 2012. ", "The fun-loving WWE superstar is already planning to make a big entrance.", "\n\n“WrestleMania is too big to put under a roof I think,” he said..\n\n“I think with WrestleMania, we have it outside. ", "We have it open. ", "I think maybe I can talk to our troops and see if I can get parachuted in off a C-130. ", "Maybe I’ll do that.”", "\n\nWhen the story broke on the Miami Herald website, fans had already begun predicting Dwayne “The Rock” Johnson’s induction in the WWE Hall of Fame. ", "Show thinks the University of Miami alum would make a great inductee, if it happens.", "\n\n“You look at the impact The Rock had on our industry from the short time he has been in the business to the megastar he is today. ", "I think if anyone deserves to be in the Hall of Fame, I’m sure The Rock does,” Show said.", "\n\n“That is a guy who is legendary and deserves every bit of his success. ", "Plus, Dwayne Johnson is one of the finest and nicest people I met in my life. ", "There are a lot of guys that you meet in this business. ", "There are people you don’t like and people you do like.", "\n\n“I have incredible respect for Dwayne and his entire family. ", "I respect the man and the character he has. ", "He’s awesome.”", "\n\nKevin Nash is someone else Show respects. ", "The two have a history dating to their World Championship Wrestling days. ", "Nash recently made an appearance at the Royal Rumble under his Diesel persona.", "\n\nThe two big men eyed each other.", "\n\n“I learned a lot from him, when I was in the nWo (new World order),” said Show. “", "When I was a greenhorn, he helped me out a lot. ", "I’m a little bit more of a seasoned veteran now, so if he wants to dance one more time as a partner or opponent, I’ll shake his hand in the middle of the ring, and we will lock horns. ", "I have no problem with that what so ever.”", "\n\nRight now Show is concentrating on the future. ", "WrestleMania 27 on April 3 from the Georgia Dome in Atlanta has not even occurred, but the veteran has already started planning for WrestleMania 28 in 2012.", "\n\n“I’m trying to recover from the Royal Rumble,” Show said. “", "We’ll see what’s going on at the Elimination Chamber and see how I fit into this year’s WrestleMania.", "\n\n“I’m excited about WrestleMania in Atlanta, but for me, personally, I’m already focusing on Miami 2012. ", "I’m going to look to be defending a championship or winning a championship in the main event.”" ]

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[ "Background\n==========\n\nInterest in the role food environments play in shaping behaviours related to food consumption and food choice has grown in recent years. ", "Researchers have often studied this relationship between individuals and their environments through creating metrics of environmental 'exposure' \\[[@B1]\\], for example neighbourhood availability of fast food outlets \\[[@B2]-[@B6]\\]. ", "However, the resulting evidence base is equivocal and the degree to which the environment determines behaviour remains unknown. ", "In terms of study design, investigations into the 'obesogenic environment' \\[[@B7]\\] are frequently large scale, quantitative, often Geographical Information Systems (GIS) based \\[[@B1],[@B8]-[@B11]\\], and importantly, rely heavily on the use of secondary data. ", "Despite this, *relatively* little is known about the accuracy of commonly used secondary food environment datasets. ", "In creating measures of food environment exposure that hope to *realistically* model individual-environment relationships, having accurate food outlet location data is critical, and so data accuracy should be better understood.", "\n\nSeveral recent studies have addressed the accuracy and reliability of secondary food outlet data sources in relation to their utility for use in health research \\[[@B12]-[@B20]\\], although most assessments have been made in the US. ", "Whilst collecting primary food outlet data might be the ideal, primary data collection is resource and time intensive. ", "There is therefore an important place for secondary data in the quantification of food environments, yet the quality and completeness of such data are not always clear. ", "In the US, companies such as Dun and Bradstreet (D&B) and InfoUSA can provide a minimal-fuss, geographically large and ready classified dataset, whilst in the UK, commercial Yellow Pages data can be purchased in bulk through providers such as Experian. ", "The use of such data represents the lowest time resource cost option for secondary data acquisition. '", "Collecting' data from local councils (governing bodies at the local level) or state departments is more complex, requiring a substantial time and resource investment to both obtain and streamline the data prior to use \\[[@B21]\\]. ", "These three types of data source ('primary', 'intensive secondary' (such as council data), and 'extensive secondary' (such as Yellow Pages or InfoUSA data)) are all potentially important, allowing accuracy to be traded for convenience where imperatives such as research timelines prevail. ", "However, in order to make the best decisions about which data to use, it is important to know how these different data sources compare.", "\n\nLake *et al.*\\[[@B12]\\] compared online and paper editions of the Yellow Pages telephone directories to the gold standard of a ground truthed food outlet database in North East England, finding positive predictive values (PPVs) of 79.1% and 82.4%, respectively. ", "Even better was the PPV for food outlet data from local councils' environmental health departments as compared with reality, at 91.5% in this area \\[[@B12]\\]. ", "In the UK, food outlets are required to register their business with local councils by law in order to facilitate routine hygiene inspections, which may explain this accuracy. ", "Other UK studies have re-iterated the accuracy of council records, reporting PPVs of 86.6% (1997) and 87.3% (2007) at two time points in Glasgow \\[[@B13]\\], and between 79-87% across urban/rural and socio-economic divides in North East England \\[[@B14]\\]. ", "The sensitivity of council data compared to 'reality' has consistently shown itself to be 'moderate' to 'excellent' \\[[@B14],[@B15]\\], according to a classification system developed by Paquet *et al.*\\[[@B16]\\]. ", "In North America, although the accuracy of state level data was questioned in one paper \\[[@B17]\\], improved PPVs and sensitivities have been found for state level food records (ground truthed data as the gold standard) as compared with the much used D&B and InfoUSA commercial datasets \\[[@B18]-[@B20]\\].", "\n\nThis said, most assessments of data validity have been made across entire study areas, not accounting for differences in completeness across socio-economic lines or urban/rural boundaries. ", "There is some suggestion that the accuracy of food outlet records may vary systematically across such divides \\[[@B14],[@B22]\\], which do exist in the UK, albeit perhaps less overtly than in the US, for example. ", "Whilst one small study in North East England did not find any significant differences in data validity by area SES or urban/rural status \\[[@B14]\\], potential differences in data integrity across these divides are important to consider as they might imbue systematic biases in downstream analyses.", "\n\nIn the UK, Ordnance Survey (OS) Points of Interest (POI) data are increasingly used in the literature as a source of information on environmental attributes such as the locations of food stores or physical activity facilities \\[[@B23]-[@B25]\\], and hold potential to be an accurate and useful source of 'extensive secondary' data due to its updateability, positional accuracy (co-ordinates are provided for environmental attributes with 1m precision), and theoretical comprehensiveness \\[[@B26]\\]; POI contains information from over 170 data suppliers, chosen for being \"the most authoritative source...for the particular type of feature they supply and for the quality and completeness of \\[their\\] data\" \\[[@B26]\\]. ", "Inaccuracies demonstrated in other sources of commercial data only enhance the appeal of POI \\[[@B12]\\], however the accuracy of these data has not yet been assessed in the published academic literature, leaving its efficacy for use in health research in question.", "\n\nUsing accurate council food outlet location data as the reference standard, this study aims to assess the validity of POI data for use in research into the (obesogenic) food environment for the first time, in Cambridgeshire, UK. ", "Reliability will be assessed as the completeness of POI records as compared to council data, which has been shown to be moderately to highly accurate in other regions of the UK, with a PPV of 91.5% in North East England \\[[@B12]\\]. ", "We aim to undertake this assessment for all POI records across the study area and to assess whether POI completeness varies by outlet type, by urban/rural status and across socio-economic divides.", "\n\nMethods\n=======\n\nFood outlet data\n----------------\n\nData on the locations of food outlets throughout Cambridgeshire, UK (Figure [1](#F1){ref-type=\"fig\"}), were sourced directly from OS under an educational license, and from local councils (n=6) throughout the region. ", "Councils were approached individually and asked to provide their *current* environmental health food outlet records under the Freedom of Information (FOI) act (for details, see <http://www.legislation.gov.uk/ukpga/2000/36/data.pdf>). ", "Both datasets were obtained in January 2012; minimising temporal mismatch between datasets was critical in making as fair a comparison as possible. ", "Duplicate records (n=5) were identified in the council food outlet records received, and removed. ", "Where no further address details were available, duplicate postcodes were assumed to represent co-existent food outlets, as postcodes usually contain multiple addresses. ", "Food outlets from both council and POI datasets were classified according to a modified 6-point food outlet classification scheme, adapted from the 21-point schema developed by Lake *et al.*\\[[@B12]\\]. ", "Any proprietary classification system already in place in the POI and council data received, was ignored. ", "Each outlet was classified only once, according to its primary trading purpose, as has been done previously \\[[@B12],[@B14]\\]. ", "Food outlets were classified using internet research, Google Street View, phone calls, and local knowledge, by a single researcher to eliminate inter-rater bias, as either: 'Café/Coffee Shops', 'Restaurants', 'Specialist Stores' (butchers, 'traditional' bakers, fishmongers and so on), 'Convenience', 'Supermarkets' (defined as belonging to a major UK supermarket chain, such as Tesco, ASDA or Sainsbury's and differentiated as such from independently owned traditional convenience stores) or 'Takeaways'. ", "These are broad categories of food outlet type, all potentially related to behaviours, as evidenced by the frequency of use of such categories in the published literature \\[[@B27]-[@B33]\\]. ", "Public houses ('pubs') were considered individually and included as 'Restaurants' only if they sold food that was more than just 'bar snacks'. ", "Mobile food outlets were excluded from the datasets as the home address of the owner was often given in lieu of the retail location.", "\n\n{#F1}\n\nOutlets were matched based on their name, address and postcode. ", "Outlets were matched, even where spelling of business name was similar but not identical, where supporting evidence (such as the same address and/or postcode) was present. ", "Food outlet locations for council and POI data were geocoded according to their postcodes and overlaid atop Lower Super Output Area (LSOA) boundaries for Cambridgeshire, using ArcGIS 10 (ESRI Inc., Redlands, CA). ", "LSOAs were attributed an urban/rural status (according to Communities and Local Government guidelines, defining small towns, villages and hamlets with fewer than 10,000 residents as 'rural' \\[[@B34]\\]), with a good mix of urban and rural areas present throughout the study area, as shown in Figure [1](#F1){ref-type=\"fig\"}. ", "LSOAs were also attributed a measure of area level socio-economic status (SES) (quintiles of Index of Multiple Deprivation (IMD) scores 2010 \\[[@B35]\\], relative to Cambridgeshire county), as also shown in Figure [1](#F1){ref-type=\"fig\"}. ", "IMD is a compound measure of SES across seven principle domains (income deprivation, employment deprivation, crime, health deprivation and disability, education, skills and training deprivation, barriers to housing and services and living environment deprivation), with scores increasing as deprivation increases \\[[@B36]\\].", "\n\nStatistical analyses\n--------------------\n\nCompleteness of POI data compared to the reference standard council data was assessed by calculating percentage agreement, positive predictive values (PPVs) and sensitivities for all outlets, and by type of food outlet, using PASW Statistics 18 (PASW Statistics Inc., Chicago, 2009). ", "These statistics have been widely employed in the literature to date \\[[@B12]-[@B14],[@B16],[@B18],[@B19]\\]. ", "Percentage agreement computes the percentage of food outlets present in both POI and council data (true positives/(true positives + false negatives + false positives)). ", "PPVs represent the percentage of outlets listed in the POI dataset that were also present in the council data (true positives/(true positives + false positives)). ", "Sensitivity represents the percentage of outlets listed in the council data that were also listed in the POI data (true positives/(true positives + false negatives)). ", "As is common in the literature, accepted sensitivity cut-offs will be applied here \\[[@B16]\\]: 'poor' \\<30%; 'fair' 31-50%; 'moderate' 51-70%; 'good' 71-90%; 'excellent' \\>91%. ", "Lake *et al.*\\[[@B12]\\] present a useful diagram showing how PPVs and sensitivities are calculated and relate to each other. ", "Differences between PPVs, sensitivities and percentage agreements for all food outlets as compared to food outlets by type were assessed using Fisher's Exact tests (preferred over chi-squared tests due to potentially small expected values). ", "PPVs and sensitivities were calculated separately for urban and rural areas and for each IMD quintile; comparisons with PPVs and sensitivities in relation to urban and least deprived reference categories were again made using Fisher's Exact tests. ", "A value of p\\<0.05 was used as the marker of statistical significance for differences.", "\n\nResults\n=======\n\nPercentage agreement\n--------------------\n\nDescriptive statistics for council and POI data received are shown in Table [1](#T1){ref-type=\"table\"}. ", "The POI data contains 524 fewer total records than were present in the council data, and fewer records by all types of food outlet, with the exception of supermarkets. ", "For cafés/coffee shop records, POI data contained 39.07% fewer gross records. ", "Table [1](#T1){ref-type=\"table\"} also shows percentage agreement between council and POI data, across all food outlets, food outlets by type, and all food outlets across urban/rural divides and SES quintiles. ", "Agreement varied according to food outlet type and was significantly different (p\\<0.05) to overall food outlet agreement (49.9%), with the exception of specialist food retailers. ", "Percentage agreement was significantly lower in rural than urban reference areas (p\\<0.001). ", "Compared to the least deprived reference areas, the third and fourth SES quintiles had significantly improved percentage agreement; other deprivation quintiles were not significantly different.", "\n\n###### \n\nDescriptive statistics and percentage agreement for all food outlets, food outlets by type, and all food outlets across urban/rural divides and socio-economic status quintiles\n\n **Food outlet category** **Council data** **POI data** **Missing POI records (%)** **Percentage agreement (%)**^**a**^ **95% CI** **95% CI for difference** \n -------------------------- ------------------ -------------- ----------------------------- ------------------------------------- ------------ --------------------------- -------------- ----------------\n **All Food Outlets** 2624 100.00 2100 100.00 19.97 49.9 (*REF*) 0.482, 0.517 *REF*\n Café/Coffee Shop 366 13.65 223 10.62 39.07 40.6\\*\\* 0.358, 0.454 0.043, 0.144\n Convenience 608 23.17 398 18.65 34.54 29.7\\*\\* 0.265, 0.330 0.166, 0.239\n Restaurant 852 32.47 757 36.05 11.15 63.5\\*\\* 0.604, 0.665 −0.171, -0.101\n Specialist Stores 248 9.45 221 10.52 10.89 47.5 0.419, 0.531 −0.033, 0.082\n Supermarket 92 3.51 93 4.43 0.00 62.3\\* 0.527, 0.712 −0.214, -0.033\n Takeaway 458 17.45 408 19.43 10.92 58.6\\*\\* 0.543, 0.628 −0.132, -0.042\n ***Urban/Rural***                \n Urban 1721 65.59 1484 70.67 13.77 52.8 (*REF*) 0.506, 0.549 *REF*\n Rural 903 34.41 616 29.33 31.78 43.0\\*\\* 0.400, 0.461 0.061, 0.134\n ***SES Quintiles***                \n SES-1 (Least Deprived) 342 13.03 234 11.14 31.58 41.2 (*REF*) 0.364, 0.461 *REF*\n SES-2 376 14.33 278 13.24 26.06 44.7 0.400, 0.494 −0.101, 0.031\n SES-3 602 22.94 552 26.29 8.31 55.0\\*\\* 0.513, 0.586 −0.198, -0.078\n SES-4 627 23.89 523 24.90 16.59 53.9\\*\\* 0.503, 0.576 −0.187, -0.068\n SES-5 (Most Deprived) 677 25.80 513 24.43 24.22 45.1 0.417, 0.486 −0.098, 0.019\n\n^a^ Significant difference (Fisher's Exact, \\*\\*p\\<0.001, \\*p\\<0.05) between food outlet category/area type and reference category (*REF*) within food outlet category/area type.", "\n\nPositive predictive value analysis\n----------------------------------\n\nAn ideal PPV would be 100%, whereby all outlets identified in the POI data were also present in the council data. ", "Table [2](#T2){ref-type=\"table\"} presents PPVs for all food outlets throughout the study area, and food outlets by type. ", "The POI data has a PPV of 74.9% overall, with PPVs ranging between 57.9-82.6% by type. ", "PPVs for Convenience and Specialist Stores and Restaurants were significantly different. ", "PPVs across urban/rural areas and SES quintiles are also presented (Table [2](#T2){ref-type=\"table\"}), and are similar to urban and the least deprived quintile reference categories.", "\n\n###### \n\nPPVs for all food outlets, food outlets by type, and all food outlets across urban/rural divides and socio-economic status quintiles\n\n **Food outlet category** **PPV (%)**^**a**^ **95% CI** **95% CI for difference**\n -------------------------- -------------------- -------------- ---------------------------\n **All Food Outlets** 74.9 (*REF*) 0.730, 0.768 *REF*\n Café/Coffee Shop 76.2 0.701, 0.817 −0.072, 0.046\n Convenience 57.9\\*\\* 0.529, 0.628 0.118, 0.222\n Restaurant 82.6\\*\\* 0.797, 0.852 −0.109, -0.043\n Specialist Stores 68.3\\* 0.618, 0.744 0.002, 0.130\n Supermarket 76.3 0.664, 0.845 −0.102, 0.074\n Takeaway 78.4 0.741, 0.823 −0.079, 0.009\n ***Urban/Rural***      \n Urban 74.6 (*REF*) 0.723, 0.768 *REF*\n Rural 74.2 0.705, 0.776 −0.037, 0.045\n ***SES Quintiles***      \n SES-1 (Least Deprived) 71.8 (*REF*) 0.656, 0.775 *REF*\n SES-2 72.7 0.670, 0.778 −0.087, 0.069\n SES-3 74.2 0.704, 0.778 −0.093, 0.044\n SES-4 77.1 0.732, 0.806 −0.121, 0.015\n SES-5 (Most Deprived) 72.1 0.680, 0.760 −0.073, 0.066\n\n^a^ Significant difference (Fisher's Exact, \\*\\*p\\<0.001, \\*p\\<0.05) between food outlet category/area type and reference category (*REF*) within food outlet category/area type.", "\n\nSensitivity analysis\n--------------------\n\nResults of sensitivity analyses are presented in Table [3](#T3){ref-type=\"table\"}, with Paquet *et al's* sensitivity cut-offs applied \\[[@B16]\\]. ", "Sensitivity for all food outlets throughout the study area was 59.9% ('moderate'), and varied, mostly significantly, according to food outlet type (as high as 77.2% for supermarkets, p\\<0.05). ", "Sensitivities were also both 'moderate' across urban/rural divides, although sensitivity in rural areas was significantly different to urban reference regions in terms of the sensitivity value proper. ", "Although sensitivity in quintile 1 of SES is described as 'fair', it is borderline 'moderate', in line with other SES quintiles. ", "This said, sensitivity values within SES quintiles 3 and 4 were significantly greater than in the most affluent reference category (p\\<0.001).", "\n\n###### \n\nSensitivity values for all food outlets, food outlets by type, and all food outlets across urban/rural divides and socio-economic status quintiles\n\n **Food outlet category** **Sensitivity (%)**^**a**^ **95% CI** **Sensitivity category**^**b**^ **95% CI for difference**\n -------------------------- ---------------------------- -------------- --------------------------------- ---------------------------\n **All Food Outlets** 59.9 (*REF*) 0.580, 0.618 Moderate *REF*\n Café/Coffee Shop 46.4\\*\\* 0.412, 0.517 Fair 0.081, 0.189\n Convenience 37.8\\*\\* 0.340, 0.418 Fair 0.178, 0.264\n Restaurant 73.4\\*\\* 0.703, 0.763 Good −0.169, -0.099\n Specialist Stores 60.9 0.545, 0.670 Moderate −0.073, 0.054\n Supermarket 77.2\\* 0.672, 0.853 Good −0.260, -0.084\n Takeaway 69.9\\*\\* 0.654, 0.740 Moderate −0.145, -0.053\n ***Urban/Rural***        \n Urban 64.6 (*REF*) 0.621, 0.666 Moderate *REF*\n Rural 50.6\\*\\* 0.473, 0.539 Moderate 0.098, 0.177\n ***SES Quintiles***        \n SES-1 (Least Deprived) 49.1 (*REF*) 0.437, 0.546 Fair *REF*\n SES-2 53.7 0.485, 0.588 Moderate −0.119, 0.027\n SES-3 67.9\\*\\* 0.640, 0.717 Moderate −0.253, -0.123\n SES-4 64.3\\*\\* 0.604, 0.680 Moderate −0.216, -0.087\n SES-5 (Most Deprived) 54.7 0.508, 0.584 Moderate −0.120, 0.010\n\n^a^ Significant difference (Fisher's Exact, \\*\\*p\\<0.001, \\*p\\<0.05) between food outlet category/area type and reference category (*REF*) within food outlet category/area type.", "\n\n^b^ Paquet *et al's s*ensitivity category cut-offs: 'poor' \\<30%; 'fair' 31-50%; 'moderate' 51-70%; 'good' 71-90%; 'excellent' \\>91%.", "\n\nDiscussion\n==========\n\nThis work examined the validity of a potentially important and increasingly used 'extensive secondary' dataset in the UK. ", "As has been noted, despite general epidemiological concern with regards to measurement accuracy \\[[@B18]\\] and the determination of exposure 'truth' \\[[@B37]\\], surprisingly little is known about the validity of commonly used secondary data sources in the field. ", "This study assessed the accuracy of POI data (at least as compared to previously validated local council records) for the first time in the published literature. ", "Although the results of this study are therefore specific to POI data, as compared with local council records in Cambridgeshire, UK, the importance of considering the validity of secondary data in these ways and across pertinent divisions remains important across all secondary datasets; this study is novel in this respect.", "\n\nIn terms of concordance between the datasets, the POI data contained 524 fewer gross records than were present in the council data, with a percentage agreement of 49.9%, translating into an overall PPV of 74.9% and sensitivity of 59.9% ('moderate'). ", "These results are largely in line with previous studies examining the accuracy of *other* secondary food environment data \\[[@B12]-[@B15],[@B18]-[@B20]\\], the caveat being that this study did not use a ground truthed dataset as a gold standard, and instead used a reliable secondary reference dataset (demonstrated to have a PPV of 91.5% in Newcastle, UK \\[[@B12]\\]) to increase the scale of the investigation.", "\n\nDifferentiation by type of food outlet revealed PPVs between 57.9% and 82.6%, with sensitivities between 37.8% ('fair') and 77.2% ('good'). ", "These assessments by food outlet type are roughly in line with those demonstrated in the literature \\[[@B12],[@B19]\\], but rather below those shown for some commercial US datasets \\[[@B18]\\]. ", "As these statistics were largely significantly sensitive to food outlet type, this research highlights the importance of considering the accuracy of secondary data for specific types of food outlet, as has been noted elsewhere \\[[@B19]\\]. ", "Although we find the lowest levels of gross completeness for cafés/coffee shops (39%), in terms of the number of missing records in POI data, convenience store records are especially incomplete with regards to percentage agreement, PPVs and sensitivity. ", "These small grocery shops are commonly cited as being 'obesogenic' \\[[@B27],[@B38],[@B39]\\], being less likely than larger supermarkets to sell 'healthful' foods \\[[@B40]\\]. ", "Given this potential gap in the POI data, this might be an area to focus on if future research is considering supplementing POI data with either council records or field work. ", "It is of note that POI appears to represent a particularly robust source of data on restaurant locations.", "\n\nImportantly, PPVs across socio-economic and urban/rural divides were similar, both to each other, and to the statistic for all outlets. ", "Such similarities have been demonstrated elsewhere \\[[@B14],[@B18]\\]. ", "For sensitivity and percentage agreement, there were exceptions, including significantly better estimates of both in some more deprived quintiles, although no evidence of a trend existed, and in urban areas. ", "This said, sensitivies across urban/rural and SES divides mostly remained 'moderate' and as such aligned with the overall sensitivity description. ", "Whilst the data should still be seen as 'imperfect' \\[[@B13]\\], some had suggested that *substantial* differences in food outlet representation across SES and urban/rural divides such as those tested here might prevail \\[[@B14],[@B22]\\], and whilst this hypothesis should be further tested in validation studies of other datasets, we do not believe this was the case here.", "\n\nThe utility of POI data may be research specific, however, if selected as a source of food outlet location data, we suggest they should be used with confidence particularly with respect to data completeness over socio-economic divides, in urban areas, and where research focuses on restaurant, supermarket or takeaway locations.", "\n\nStrengths of this study include the fair comparison of contemporaneous datasets, the application of a 6 category food outlet classification scheme whose outlet types should relate directly to future deductive research, and its large geographical scale, which enabled an assessment of over 2000 food outlets in each dataset. ", "In particular, using established statistics (percentage agreement, PPVs and sensitivies) across urban/rural and socio-economic divides allowed an assessment of the likelihood of systematic geographical differences in completeness. ", "To our knowledge, this is the first time that such an appraisal has been made in the published literature on a large scale.", "\n\nThere were several key limitations to this study. ", "In order to enable the large study area, field work was not conducted, choosing instead to use local council data as our 'gold (reference) standard'. ", "Local council data have been shown accurate in several other regions of the UK, however they are unlikely to be complete, resulting in a potential lack of comparability with previous studies that can relate directly to the food environment *reality*. ", "Despite this limitation, the strength of results found here suggest that if council data are indeed less complete than we might hope, or are systematically incomplete (for example, across socio-economic divides) they are at least aligned in these respects with POI records. ", "In order to maximise heterogeneity in socio-economic status throughout the study area, quintiles of SES were calculated relative to the study area only. ", "Increased sensitivity in detecting SES differences between LSOAs was useful for these analyses, however, our findings may not be applicable to the most deprived locales, which are substantially under-represented throughout Cambridgeshire (IMD scores are positively skewed towards being lower (less deprived); mean IMD for Cambridgeshire=15.51 (SD=11.44), range of possible IMD scores for England as a whole 0.53-87.80). ", "This potential limitation may lead to a lesser degree of generalisability outside this study area, however it does not compromise the accuracy of *these* results. ", "To facilitate a fair comparison of the datasets, we attempted to obtain as contemporaneous information as possible. ", "We asked OS and local councils for current data in January 2012 to facilitate this, however, it is possible that either dataset may not reflect the food environment at precisely the same time. ", "Whilst some exclusions in the datasets were made based on food not sold directly to the public (food producers, for example), exclusions of market traders or mobile food stands were made predominantly because addresses were for the traders' home addresses and not the retail sites themselves. ", "These types of food retailers are likely important sources of food \\[[@B14],[@B22]\\], potentially with a socio-economic gradient of use \\[[@B41],[@B42]\\], and should be considered where possible in future validation work.", "\n\nIn terms of the POI dataset itself, the data were not without duplicates that needed to be found and removed (n=105). ", "The classification system supplied was too general to be of real use in most health research (for details see, <http://www.ordnancesurvey.co.uk/oswebsite/docs/product-schemas/points-of-interest-classifications-scheme.pdf>) so a project specific classification scheme such as the one used here would almost certainly be required. ", "POI contains records beyond simply the foodscape, making it difficult to discern whether listed establishments sold food or not. ", "In council datasets, outlets are listed *precisely because* they sell food. ", "This breadth may lead to the omission of important sources of food within the environment, for example from pharmacies, such as Boots the Chemist, a national chain that often but not always sells food items. ", "Investigative work would be required when using POI data to determine whether or not each of these individual stores sells food.", "\n\nConclusions\n===========\n\nAccurate analysis in health and policy research begins with accurate data. ", "Ordnance Survey Points of Interest records generally compared favourably here in relation to data from local councils' environmental health departments. ", "We observed few notable systematic variations in POI completeness (PPV/sensitivity) over urban/rural and SES divides, however when type of outlet was considered, convenience stores appeared to be the least well represented in the POI, and consideration must therefore be given to the types of outlets being studied when selecting a dataset.", "\n\nThe utility of POI is boosted when its relative ease of acquisition is considered (in relation to both 'intensive secondary' council data, and primary data collection). ", "However, this is not to say that by *combining* POI data with local council data, one might be able to build an even more accurate picture of the food environment. ", "Future research using a ground truthed dataset over an equivalent study area is necessary to ascertain whether this is likely to be the case.", "\n\nCompeting interests\n===================\n\nThe authors declare they have no completing interests.", "\n\nAuthors' contributions\n======================\n\nThe study design was jointly devised by TB and FH. ", "TB was responsible for data collection from local councils, FH for data acquisition from Ordnance Survey. ", "TB led on data analysis. ", "TB and FH drafted the manuscript together. ", "Both authors read and approved the final manuscript.", "\n\nAcknowledgements\n================\n\nThis work was undertaken by the Centre for Diet and Activity Research (CEDAR), a UK Clinical Research Collaboration (UKCRC) Public Health Research Centre of Excellence. ", "Funding from the British Heart Foundation, Economic and Social Research Council, Medical Research Council, the National Institute for Health Research and the Wellcome Trust under the auspices of the UK Clinical Research Collaboration, is gratefully acknowledged. ", "The digital maps used hold Crown Copyright from EDINA Digimap, a JISC supplied service. ", "We are grateful to Cambridgeshire local councils and Ordnance Survey for kindly supplying data to enable this work.", "\n" ]

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[ "Tuesday, 27 July 2010\n\nThis decorative technique joins two pieces of fabric or lace leaving a space in between to allow for stitches. ", "The fabric edges may be corded or folded. ", "This technique also works well when attaching decorative lace to fabric.", "\n\nThese distort real objects. ", "In this natural designs are simplified, exaggerated, rearranged or even distorted to achieve the purpose of the design. ", "In textile industry, patterns of richly coloured, delicately petaled roses and patterns of rose’s sharp thorns are both referred to as floral. ", "The floral category includes all the gatherings of the flower garden, in fact including grasses, but agricultural produce like fruit and vegetables is considered a conversational subject. ", "Floral motifs are more common in women’s clothes and furnishing materials. ", "Examples of floral patterns are\n\na) All over non-directional: The motifs of the patterns cover more than 50 % of the field. ", "Such layouts are popular with textile and fashion designers because they tend to disguise a pattern repeat.", "\n\nb) Allover set: It is a formal layout of grids and diagonals.", "\n\nc) Baskets, flowerpots and vases: These motifs help to organize a floral pattern, providing a visual focus. ", "Flower spilling out of basket can have a romantic, uplifting effect. ", "Flowerpots and vases resolve the issue of how to deal with plants untidy roots and stems.", "\n\nd) Bouquets and nosegays: These are popular with designers because they offer a way to combine a variety of flowers in one motif. ", "They evoke images of romance, giving gifts- and sentiments of spring.", "\n\nGeometric designs:\n\nThese are based on pure forms of the circle, rectangle and triangles etc. ", "Geometric motifs include stripes, dots, checks, and plaids as well as many less usual forms. ", "A geometric is an abstract or non-representational motif, a shape that is not a picture of something out in the real world. ", "Examples of geometric designs are basket weave design, check board, chevron and herringbone weave, diagonal stripes, diaper pattern, ogee pattern, plaids and polka dots\n\nAbstract designs:\n\nThese have little or no reference to real object. ", "Abstract implies an element of impression and a greater freedom than is found in most geometric designs. ", "This type of design is used in modern art.", "\n\nEthnic designs:\n\nMost of the patterns illustrated in this chapter however were produced not for the overseas market but for the domestic one. ", "Ethnic fashions – meaning in textile vocabulary, any pattern or style with a foreign or exotic feeling – come and go in western design. ", "The ethnic styles have their day in western design and then submerge, to reappear Whenever some one senses that the time is right for a new twist on the constant demand for a folkloric pattern. ", "Examples are American Indian look, batik and Indonesian look, Egyptian look, Mexican design and paisley, tie and dye patterns.", "\n\nArt movements and period styles:\n\nDesigns like these are often the links between cloth and the fine arts- they are part of movements and ideas of the age, and are moved to carry theme through in textiles. ", "Examples are Jacobean looks and super graphics\n\nFor each of these ways one can interpret the designs based on one of the following crieteria:\n\nMotif: the most important factor in any design, determines the family to which the pattern belongs. ", "This is the basic image –a rose, a square, a clown a paisley\n\nLayout: It describes the arrangement of the motif- whether it is spaced widely or closely on the ground, in neat order or apparently at random, or in rows that form stripes.", "\n\nColour: designs are so classified when a particular dye- indigo, madder or Turkey red say –is the strongest element of their look.", "\n\nPlain golden or silver wires are called ‘Badla’ and when these wires are wound around a thread they are called ‘Kasab’, spangles of metal are ‘Sitara’ and tiny dots made out of badla are called ‘Mukaish’. ", "The days of using real gold or silver are now history, what you now get is synthetic\n\nThursday, 22 July 2010\n\nSoof (lit. “", "triangle”) is a technique counted out from the back of the fabric. ", "Geometrical, whimsical variations of traditional motifs, including stylized flowers and figures, convey wishes of fertility and prosperity for the newlyweds. ", "The designs are created with surface-satin stitches of varying lengths that allow most of the valuable silk to be visible on the front.", "\n\nSometimes back I have written an article on microspinning. ", "I have also mentioned how at Chirala in Andhra Pradesh, a fabric by the name of Malkha Cotton is being produced from microspinning.", "\n\nI am glad that the Malkha fabric has become one of the choices of the designers and caught the attention of the media so-much-so that an article in Mint is devoted to this fabric. ", "The source of the image is website of Malkha.", "\n\nThe technology of spinning is hailed as Decentralised Cotton Yarn making-essentially using the same technology from Vortex as that of microspinning.", "\n\nNow that you've finished reading this post, what are you going do? ", "You should go join the Forum.", "\n\nSunday, 4 July 2010\n\nI need a solution to control the skew and spirality of single jersey 160g with 30/s. Please provide me with some solution\n\nSpirality is a dimensional distortion in circular knitted fabric. ", "Spirality is bad as it leads to displacement of seams and mismatched patterns. ", "It can also leads to sewing difficulties. ", "The major cause of spirality is the twist in roving and yarn. ", "To minimize spirality, Z-twist yarns should be knotted on clockwise rotating machine and S-twist yarns on counter clockwise rotating machines. ", "Tighter Fabics exhibit less spirality compared to looser fabrics which means that finer gauge machines will reduce spirality.", "\n\nIt can also be reduced by setting the twist either by autoclave treatment, yarn dyeing or using balanced plied yarns.", "\n\nThursday, 1 July 2010\n\nIt is a traditional embroidery of the state of Karnataka and is done by counting the threads of the warp and weft with the designs traced or implanted as outlines.", "\n\nThere are essentially four types of kasuti embroidery. ", "They are Neyge Kasuti, Murgi Kasuti, Menthya Kasuti and Gavanthi Kasuti. ", "Gavanti and Murgi comprise a straight and zig zag stitch. ", "The stitch is identical on both sides of the cloth where as Menthe and Negi Stitch are dissimilar on either side of the fabric. ", "The Difference between Menthe and Negi Stitch is that while the Menthe stitch resembles a cross-stitch, the Negi stitch provides a woven style. ", "These stitches are dissimilar on either side of the fabric.", "\n\nThe Basis of Gavanti and Murgi Stitich is Hoblein Stitich. ", "To understand the concept of Hoblein stitch, you can visit this site.", "\n\nExcellent Material on Kasuti Embroidery is available in this document.", "\n\nAn equally amazing tutorial on Kasuti Embroidery is available in this blog and in this blog. ", "Another tutorial is available here in the same blog.", "\n\nNow that you've finished reading this post, what are you going do? ", "You should go join the Forum." ]

{ "pile_set_name": "Pile-CC" }

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[ "OpenOffice.org - the free and open productivity suitehttp://www.openoffice.org/10316af0cc804051cd9a6b1237f94667f9e4a9ec9762686542de58944d3a3b8f04d2011e93a450578d77a5983beef32860347ba10d4afadacbc63ff9febf217ae86e6d347c9599ec9762686542de58944d3a3b8f04d2011e93a45http://openoffice.cs.utah.edu/contrib/dictionaries/md5sums.txthttp://www.mirrorservice.org/sites/download.openoffice.org/contrib/dictionaries/md5sums.txthttp://ftp.nluug.nl/pub/office/openoffice/contrib/dictionaries/md5sums.txthttp://ftp.snt.utwente.nl/pub/software/openoffice/contrib/dictionaries/md5sums.txthttp://ftp-stud.hs-esslingen.de/pub/Mirrors/ftp.openoffice.org/contrib/dictionaries/md5sums.txthttp://ftp5.gwdg.de/pub/openoffice/contrib/dictionaries/md5sums.txthttp://ftp3.gwdg.de/pub/openoffice/contrib/dictionaries/md5sums.txthttp://ftp4.gwdg.de/pub/openoffice/contrib/dictionaries/md5sums.txthttp://ooo.mirror.garr.it/mirrors/openoffice/contrib/dictionaries/md5sums.txthttp://ftp.tu-chemnitz.de/pub/openoffice-extended/contrib/dictionaries/md5sums.txtftp://ftp.man.poznan.pl/pub/openoffice/contrib/dictionaries/md5sums.txt" ]

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[ "#import \"GPUImageTwoPassFilter.h\"\n\n@implementation GPUImageTwoPassFilter\n\n#pragma mark -\n#pragma mark Initialization and teardown\n\n- (id)initWithFirstStageVertexShaderFromString:(NSString *)firstStageVertexShaderString firstStageFragmentShaderFromString:(NSString *)firstStageFragmentShaderString secondStageVertexShaderFromString:(NSString *)secondStageVertexShaderString secondStageFragmentShaderFromString:(NSString *)secondStageFragmentShaderString;\n{\n if (!(", "self = [super initWithVertexShaderFromString:firstStageVertexShaderString fragmentShaderFromString:firstStageFragmentShaderString]))\n {\n\t\treturn nil;\n }\n \n secondProgramUniformStateRestorationBlocks = [NSMutableDictionary dictionaryWithCapacity:10];\n\n runSynchronouslyOnVideoProcessingQueue(^{\n [GPUImageContext useImageProcessingContext];\n\n secondFilterProgram = [[GPUImageContext sharedImageProcessingContext] programForVertexShaderString:secondStageVertexShaderString fragmentShaderString:secondStageFragmentShaderString];\n \n if (!", "secondFilterProgram.initialized)\n {\n [self initializeSecondaryAttributes];\n \n if (![", "secondFilterProgram link])\n {\n NSString *progLog = [secondFilterProgram programLog];\n NSLog(@\"Program link log: %@\", progLog);\n NSString *fragLog = [secondFilterProgram fragmentShaderLog];\n NSLog(@\"Fragment shader compile log: %@\", fragLog);\n NSString *vertLog = [secondFilterProgram vertexShaderLog];\n NSLog(@\"Vertex shader compile log: %@\", vertLog);\n secondFilterProgram = nil;\n NSAssert(NO, @\"Filter shader link failed\");\n }\n }\n \n secondFilterPositionAttribute = [secondFilterProgram attributeIndex:@\"position\"];\n secondFilterTextureCoordinateAttribute = [secondFilterProgram attributeIndex:@\"inputTextureCoordinate\"];\n secondFilterInputTextureUniform = [secondFilterProgram uniformIndex:@\"inputImageTexture\"]; // This does assume a name of \"inputImageTexture\" for the fragment shader\n secondFilterInputTextureUniform2 = [secondFilterProgram uniformIndex:@\"inputImageTexture2\"]; // This does assume a name of \"inputImageTexture2\" for second input texture in the fragment shader\n \n [GPUImageContext setActiveShaderProgram:secondFilterProgram];\n \n glEnableVertexAttribArray(secondFilterPositionAttribute);\n glEnableVertexAttribArray(secondFilterTextureCoordinateAttribute);\n });\n\n return self;\n}\n\n- (id)initWithFirstStageFragmentShaderFromString:(NSString *)firstStageFragmentShaderString secondStageFragmentShaderFromString:(NSString *)secondStageFragmentShaderString;\n{\n if (!(", "self = [self initWithFirstStageVertexShaderFromString:kGPUImageVertexShaderString firstStageFragmentShaderFromString:firstStageFragmentShaderString secondStageVertexShaderFromString:kGPUImageVertexShaderString secondStageFragmentShaderFromString:secondStageFragmentShaderString]))\n {\n\t\treturn nil;\n }\n \n return self;\n}\n\n- (void)initializeSecondaryAttributes;\n{\n [secondFilterProgram addAttribute:@\"position\"];\n\t[secondFilterProgram addAttribute:@\"inputTextureCoordinate\"];\n}\n\n#pragma mark -\n#pragma mark Managing targets\n\n- (GPUImageFramebuffer *)framebufferForOutput;\n{\n return secondOutputFramebuffer;\n}\n\n- (void)removeOutputFramebuffer;\n{\n secondOutputFramebuffer = nil;\n}\n\n#pragma mark -\n#pragma mark Rendering\n\n- (void)renderToTextureWithVertices:(const GLfloat *)vertices textureCoordinates:(const GLfloat *)textureCoordinates;\n{\n if (self.preventRendering)\n {\n [firstInputFramebuffer unlock];\n return;\n }\n \n [GPUImageContext setActiveShaderProgram:filterProgram];\n \n outputFramebuffer = [[GPUImageContext sharedFramebufferCache] fetchFramebufferForSize:[self sizeOfFBO] textureOptions:self.outputTextureOptions onlyTexture:NO];\n [outputFramebuffer activateFramebuffer];\n \n [self setUniformsForProgramAtIndex:0];\n \n glClearColor(backgroundColorRed, backgroundColorGreen, backgroundColorBlue, backgroundColorAlpha);\n glClear(GL_COLOR_BUFFER_BIT);\n \n\tglActiveTexture(GL_TEXTURE2);\n\tglBindTexture(GL_TEXTURE_2D, [firstInputFramebuffer texture]);\n\t\n\tglUniform1i(filterInputTextureUniform, 2);\n \n glVertexAttribPointer(filterPositionAttribute, 2, GL_FLOAT, 0, 0, vertices);\n\tglVertexAttribPointer(filterTextureCoordinateAttribute, 2, GL_FLOAT, 0, 0, textureCoordinates);\n \n glDrawArrays(GL_TRIANGLE_STRIP, 0, 4);\n \n [firstInputFramebuffer unlock];\n firstInputFramebuffer = nil;\n \n // This assumes that any two-pass filter that says it desires monochrome input is using the first pass for a luminance conversion, which can be dropped\n// if (!", "currentlyReceivingMonochromeInput)\n// {\n // Run the first stage of the two-pass filter\n// [super renderToTextureWithVertices:vertices textureCoordinates:textureCoordinates];\n// }\n\n // Run the second stage of the two-pass filter\n secondOutputFramebuffer = [[GPUImageContext sharedFramebufferCache] fetchFramebufferForSize:[self sizeOfFBO] textureOptions:self.outputTextureOptions onlyTexture:NO];\n [secondOutputFramebuffer activateFramebuffer];\n [GPUImageContext setActiveShaderProgram:secondFilterProgram];\n if (usingNextFrameForImageCapture)\n {\n [secondOutputFramebuffer lock];\n }\n\n [self setUniformsForProgramAtIndex:1];\n \n glActiveTexture(GL_TEXTURE3);\n glBindTexture(GL_TEXTURE_2D, [outputFramebuffer texture]);\n glVertexAttribPointer(secondFilterTextureCoordinateAttribute, 2, GL_FLOAT, 0, 0, [[self class] textureCoordinatesForRotation:kGPUImageNoRotation]);\n\n // TODO: Re-enable this monochrome optimization\n// if (!", "currentlyReceivingMonochromeInput)\n// {\n// glActiveTexture(GL_TEXTURE3);\n// glBindTexture(GL_TEXTURE_2D, outputTexture);\n// glVertexAttribPointer(secondFilterTextureCoordinateAttribute, 2, GL_FLOAT, 0, 0, [[self class] textureCoordinatesForRotation:kGPUImageNoRotation]);\n// }\n// else\n// {\n// glActiveTexture(GL_TEXTURE3);\n// glBindTexture(GL_TEXTURE_2D, sourceTexture);\n// glVertexAttribPointer(secondFilterTextureCoordinateAttribute, 2, GL_FLOAT, 0, 0, textureCoordinates);\n// }\n \n\tglUniform1i(secondFilterInputTextureUniform, 3);\n \n glVertexAttribPointer(secondFilterPositionAttribute, 2, GL_FLOAT, 0, 0, vertices);\n\n glClearColor(0.0f, 0.0f, 0.0f, 1.0f);\n glClear(GL_COLOR_BUFFER_BIT);\n\n glDrawArrays(GL_TRIANGLE_STRIP, 0, 4);\n [outputFramebuffer unlock];\n outputFramebuffer = nil;\n \n if (usingNextFrameForImageCapture)\n {\n dispatch_semaphore_signal(imageCaptureSemaphore);\n }\n}\n\n- (void)setAndExecuteUniformStateCallbackAtIndex:(GLint)uniform forProgram:(GLProgram *)shaderProgram toBlock:(dispatch_block_t)uniformStateBlock;\n{\n// TODO: Deal with the fact that two-pass filters may have the same shader program identifier\n if (shaderProgram == filterProgram)\n {\n [uniformStateRestorationBlocks setObject:[uniformStateBlock copy] forKey:[NSNumber numberWithInt:uniform]];\n }\n else\n {\n [secondProgramUniformStateRestorationBlocks setObject:[uniformStateBlock copy] forKey:[NSNumber numberWithInt:uniform]];\n }\n uniformStateBlock();\n}\n\n- (void)setUniformsForProgramAtIndex:(NSUInteger)programIndex;\n{\n if (programIndex == 0)\n {\n [uniformStateRestorationBlocks enumerateKeysAndObjectsUsingBlock:^(id key, id obj, BOOL *stop){\n dispatch_block_t currentBlock = obj;\n currentBlock();\n }];\n }\n else\n {\n [secondProgramUniformStateRestorationBlocks enumerateKeysAndObjectsUsingBlock:^(id key, id obj, BOOL *stop){\n dispatch_block_t currentBlock = obj;\n currentBlock();\n }];\n }\n}\n\n@end\n" ]

{ "pile_set_name": "Github" }

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[ "Validity and reliability of the Persian epilepsy quality of life questionnaire.", "\nTo translate and test the reliability and validity of the 31-item epilepsy quality of life questionnaire (QOLIE-31) in Iranian epileptic patients. ", "In order to standardize the questionnaire, using a standard \"forward-backward\" translation, cognitive debriefing, and cultural adaptation procedure, the English version of the QOLIE-31 was translated to Persian (the Iranian official language). ", "The subjects were Epileptic patients, over 18 years old, referred to the Motaharri Clinic, Shiraz, Southern Iran from March 2007 to March 2008. ", "The reliability and internal consistency of the questionnaire were assessed by Cronbach`s alpha coefficient. ", "Validity was assessed using convergent and disconvergent validity through Spearman`s correlation. ", "Two hundred and eleven epileptic patients (88 females [41.7%], 123 males [58.3%]), with a mean ± SD age of 28.7±11.6 years were enrolled in the study. ", "Reliability analysis showed a satisfactory result (Cronbach's α=0.890). ", "Internal consistency was satisfactory for both demographic and patients' clinical characteristics (α≥0.70). ", "The scaling success rates were 100% for convergent validity of each scale. ", "Disconvergent validity for all 7 scales was good. ", "The Persian version of the QOLIE-31 questionnaire has good structural characteristics, is a reliable and valid instrument, and can be used for measuring the effect of epilepsy on the quality of life." ]

{ "pile_set_name": "PubMed Abstracts" }

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[ "Game engine technologies have come a long way since the 8-bit days, and thanks to consoles like the Xbox 360 as well as the PS3, gamers can now enjoy them from the comfort of their own home. ", "Continue reading to see five amazing game technologies.", "\n\n1. ", "Unreal Engine 3\n\nDemonstrated at the 2011 Game Developers Conference, this real-time demo of ‘Samaritan’ shows what Epic wants to see in the next generation of games; the features demonstrated in this video are available now in UDK.", "\n\n2. ", "VFX Tech Demo\n\nWe have seen the future of facial animations in games, and it’s called VFX. ", "It’s a piece of technology by the guys at Janimation and could almost be considered too realistic, but we’ll leave that up to you to decide.", "\n\n3. ", "From Dust\n\nDirect from Gamescom, Ubisoft gives us a first look at From Dust, a new game from Eric Chahi featuring an exotic world that can be shaped at will by your divine powers. ", "The most impressive thing about this title is definitely its game engine.", "\n\n4. ", "NVIDIA Hair\n\nThis NVIDIA tech demo, shown at GDC 2010, simulates 18,000 individual strands of human hair simultaneously, completely interactive with light, wind, and, apparently, conditioning – just look at that bounce and shine.", "\n\n5. ", "CryEngine 3\n\nThis followup to CryEngine 2 will supposedly feature many advanced graphical, physical and animation technologies as well as many gameplay enhancements. ", "Best of all, it can be used for PC as well as console games." ]

{ "pile_set_name": "OpenWebText2" }

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[ "Q:\n\nUndefined index message in drupal\n\nWhen editing a view, I get the following message in the popup.", "\n\nNotice: Undefined index: advertising_block in eva_plugin_display_entity->options_summary() \n (line 41 of/.../sites/all/modules/eva/eva_plugin_display_entity.inc).", "\n\nThis view contains 2 displays: a view attached and a block (called advertising_block). ", "This error message appeared both before and after deleting the \"advertising_block\" to which it seems to refer.", "\nAny idea of what may be causing this error message and how to fix it?", "\n\nA:\n\nBasically, the EVA module is getting called with a bundle 'advertising_block' that it doesn't know how to handle. ", "If you used to have a block named 'advertising_block', that would be a suspect. ", "\nThe way to solve this is to (with devel installed) use the approach in webchick's Quick and Dirty Debugging. ", "Right before line 41 of/.../sites/all/modules/eva/eva_plugin_display_entity.inc, make a temporary hack.", "\nFirst, install and enable the devel module.", "\nThe code there is:\n foreach ($this->get_option('bundles') as $bundle) {\n $bundle_names[] = $entity_info['bundles'][$bundle]['label'];\n }\n\nso we'll change it to\n foreach ($this->get_option('bundles') as $bundle) {\n if ($bundle == 'advertising_block') {\n dpm(debug_backtrace(), 'Bundle advertising_block being requested');\n }\n $bundle_names[] = $entity_info['bundles'][$bundle]['label'];\n }\n\nNow when we hit that page, we'll see the backtrace (who called this thing, maybe why), and will have a path forward.", "\nWhen you're through with this debugging code, don't forget to revert to the original code and disable devel module.", "\nAnd remember that debugging like this should be done on a local dev site, not on production. ", "Some more strategies are at randyfay.com/debugging\n\n" ]

{ "pile_set_name": "StackExchange" }

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[ "Isfinder\n\nISfinder is a database of insertion sequences isolated from eubacteria and archae.", "\n\nSee also\n Mobile genetic elements\n\nReferences\n\nExternal links\n http://www-is.biotoul.fr\n\nCategory:Biological databases\nCategory:Mobile genetic elements" ]

{ "pile_set_name": "Wikipedia (en)" }

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[ "Q:\n\nButton Click event fires twice, even with ClickMode set to \"Press\"\n\nA XAML Button's Click even fires twice with a left-button mouse click.", "\nI added the XAML Button Property ClickMode = \"Press\", but that made no difference. ", " The Microsoft documentation for ClickMode indicates that that should have solved the problem.", "\nI discovered this with a breakpoint in the C++ code for the Click event handler, which stopped the code twice for each button click.", "\nXAML code within a Grid on a Page container:\n <Button x:Name=\"button_listFiles\" Grid.", "Column=\"1\" Grid.", "Row=\"0\" Content=\"List\" HorizontalAlignment=\"Center\" \n Click=\"button_listFiles_Click\" ClickMode=\"Press\"></Button>\n\nC++ Click event handler:\n void MainPage::button_listFiles_Click(IInspectable const& sender, RoutedEventArgs const& e) {\n\n MainPage::counter++;\n\n textBox_directoryName().Text(to_hstring(counter));\n }\n\nThe MainPage::counter++; counts the number of times the handler is called and textBox_directoryName().Text(to_hstring(counter)); displays the result on the XAML page, used for attempted debugging.", "\n\nA:\n\nJust to close this out, the answer is the several comments between me and Kenny Kerr.", "\n\n" ]

{ "pile_set_name": "StackExchange" }

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[ "using System;\nusing System.", "Collections.", "Generic;\nusing System.", "IO;\nusing System.", "Linq;\nusing System.", "Text;\nusing Microsoft.", "Extensions.", "Logging;\nusing Microsoft.", "Extensions.", "Options;\nusing Newtonsoft.", "Json;\nusing VirtoCommerce.", "Platform.", "Core.", "Common;\nusing VirtoCommerce.", "Platform.", "Core.", "Modularity;\nusing VirtoCommerce.", "Platform.", "Modules;\nusing VirtoCommerce.", "Platform.", "Modules.", "External;\nusing Xunit;\nusing Xunit.", "Extensions.", "Ordering;\n\nnamespace VirtoCommerce.", "Platform.", "Tests.", "Modularity\n{\n //the Order need for saparate runing UnitTests where use static Platform.", "CurrentVersion\n [Collection(\"Modularity\"), Order(1)]\n public class ExternalModuleCatalogTests\n {\n [Fact]\n public void PublishNewVersionTest()\n {\n //Arrange\n var v3_0_0_beta1 = new ModuleManifest\n {\n Version = \"3.0.0\",\n VersionTag = \"beta1\",\n PlatformVersion = \"3.0.0\"\n };\n var v3_0_0 = new ModuleManifest\n {\n Version = \"3.0.0\",\n PlatformVersion = \"3.0.0\"\n };\n var v3_1_0_beta1 = new ModuleManifest\n {\n Version = \"3.1.0\",\n VersionTag = \"beta1\",\n PlatformVersion = \"3.0.0\"\n };\n var v3_1_0_beta2 = new ModuleManifest\n {\n Version = \"3.1.0\",\n VersionTag = \"beta2\",\n PlatformVersion = \"3.0.0\"\n };\n var v3_1_0 = new ModuleManifest\n {\n Version = \"3.1.0\",\n PlatformVersion = \"3.0.0\"\n };\n\n var extModuleManifest = new ExternalModuleManifest\n {\n Id = \"A\"\n };\n\n //Act\n extModuleManifest.", "PublishNewVersion(v3_0_0_beta1);\n //Assert\n Assert.", "True(extModuleManifest.", "Versions.", "Count() == 1);\n Assert.", "True(extModuleManifest.", "Versions.", "Contains(ExternalModuleManifestVersion.", "FromManifest(v3_0_0_beta1)));\n\n //Act\n extModuleManifest.", "PublishNewVersion(v3_0_0);\n //Assert\n Assert.", "True(extModuleManifest.", "Versions.", "Count() == 2);\n Assert.", "True(extModuleManifest.", "Versions.", "Contains(ExternalModuleManifestVersion.", "FromManifest(v3_0_0)));\n\n //Act\n extModuleManifest.", "PublishNewVersion(v3_1_0_beta1);\n //Assert\n Assert.", "True(extModuleManifest.", "Versions.", "Count() == 2);\n Assert.", "True(extModuleManifest.", "Versions.", "Contains(ExternalModuleManifestVersion.", "FromManifest(v3_0_0)));\n Assert.", "True(extModuleManifest.", "Versions.", "Contains(ExternalModuleManifestVersion.", "FromManifest(v3_1_0_beta1)));\n\n //Act\n extModuleManifest.", "PublishNewVersion(v3_1_0_beta2);\n //Assert\n Assert.", "True(extModuleManifest.", "Versions.", "Count() == 2);\n Assert.", "True(extModuleManifest.", "Versions.", "Contains(ExternalModuleManifestVersion.", "FromManifest(v3_0_0)));\n Assert.", "True(extModuleManifest.", "Versions.", "Contains(ExternalModuleManifestVersion.", "FromManifest(v3_1_0_beta2)));\n\n //Act\n extModuleManifest.", "PublishNewVersion(v3_1_0);\n //Assert\n Assert.", "True(extModuleManifest.", "Versions.", "Count() == 2);\n Assert.", "True(extModuleManifest.", "Versions.", "Contains(ExternalModuleManifestVersion.", "FromManifest(v3_1_0)));\n }\n\n [Theory]\n [InlineData(\"2.12.0\", \"1.4.0\")]\n [InlineData(\"3.1.0\", \"2.0.0\")]\n public void CreateDirectory_CreateTestDirectory(string platformVersion, string effectiveModuleVersion)\n {\n //Arrange\n PlatformVersion.", "CurrentVersion = SemanticVersion.", "Parse(platformVersion);\n var modules = new[]\n {\n new ExternalModuleManifest\n {\n Id = \"A\",\n Versions = new []\n {\n new ExternalModuleManifestVersion\n {\n Version = \"1.5.0\",\n PlatformVersion = \"2.14.0\"\n },\n new ExternalModuleManifestVersion\n {\n Version = \"1.4.0\",\n PlatformVersion = \"2.12.0\"\n },\n new ExternalModuleManifestVersion\n {\n Version = \"1.3.0\",\n PlatformVersion = \"2.12.0\"\n },\n new ExternalModuleManifestVersion\n {\n Version = \"2.0.0\",\n PlatformVersion = \"3.0.0\"\n },\n }\n }\n };\n\n //Act\n var extCatalog = CreateExternalModuleCatalog(modules);\n extCatalog.", "Load();\n\n //Assert\n var module = extCatalog.", "Modules.", "FirstOrDefault() as ManifestModuleInfo;\n Assert.", "NotNull(module);\n Assert.", "Equal(SemanticVersion.", "Parse(effectiveModuleVersion), module.", "Version);\n }\n\n private static ExternalModuleCatalog CreateExternalModuleCatalog(ExternalModuleManifest[] manifests)\n {\n var localModulesCatalog = new Moq.", "Mock<ILocalModuleCatalog>();\n localModulesCatalog.", "Setup(x => x.Modules).Returns(new List<ManifestModuleInfo>());\n var json = JsonConvert.", "SerializeObject(manifests);\n var client = new Moq.", "Mock<IExternalModulesClient>();\n client.", "Setup(x => x.OpenRead(Moq.", "It.", "IsAny<Uri>())).Returns(new MemoryStream(Encoding.", "UTF8.GetBytes(json ?? \"\")));", "\n var logger = new Moq.", "Mock<ILogger<ExternalModuleCatalog>>();\n\n var options = Options.", "Create(new ExternalModuleCatalogOptions());\n var result = new ExternalModuleCatalog(localModulesCatalog.", "Object, client.", "Object, options, logger.", "Object);\n return result;\n }\n\n }\n}\n" ]

{ "pile_set_name": "Github" }

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[ "Earthmoving Machinery\n\nRodwell Movez Earth (earthmoving) has the right machines & the expertise to ensure that every earthmoving / excavation job is carried out to a high standard to enhance your property from basic earthworks to a unique landscape design" ]

{ "pile_set_name": "Pile-CC" }

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[ "This section is intended to provide a background to the various embodiments of the technology described in this disclosure. ", "The description in this section may include concepts that could be pursued, but are not necessarily ones that have been previously conceived or pursued. ", "Therefore, unless otherwise indicated herein, what is described in this section is not prior art to the description and/or claims of this disclosure and is not admitted to be prior art by the mere inclusion in this section.", "\nCurrently, wireless communication networks or systems, such as MilliMeter-Wave (MMW) wireless systems, operating at high frequencies from 30-300 GHz, are emerging as a promising technology to meet exploding bandwidth requirements by enabling multi-Gb/s speeds. ", "For example, the 5th Generation (5G) network is likely to be a combination of evolved 3rd Generation (3G) technologies, the 4th Generation (4G) technologies and emerging or substantially new components such as Ultra-Density Network (UDN), which is also referred to as MMW Radio Access Technology (RAT). ", "At such high frequencies, a large number of antennas can be available at a transmitter, a receiver, or both. ", "In order to make up for the large propagation loss that typically occurs, beam-forming becomes a very important feature in MMW wireless systems.", "\nBeam-forming is a signal processing technique used for directional signal transmission and/or reception. ", "For Transmitter (TX) beamforming, the signals are concentrated in the desired direction via applying a selected precoding vector for the TX antenna array. ", "For Receiver (RX) beamforming, the RX beam of the receiver antennas are concentrated in the incoming direction of the radio signals by applying a selected precoding vector for the RX antenna array. ", "Beam-forming can be used at both the transmitting and receiving ends in order to achieve spatial selectivity. ", "The improvement compared with omnidirectional reception/transmission is known as a beam-forming gain. ", "When multiple antennas are available at transmitters, receivers or both, it is therefore important to apply efficient beam patterns to the antennas to better exploit the spatial selectivity of the corresponding wireless channel.", "\nFIG. ", "1 schematically shows one example MMW RAT network. ", "As shown in FIG. ", "1, there is a network node or a control node called as Central Control Unit (CCU), which is at least responsible for parameter configurations and coordination among Access Nodes (ANs), e.g., AN1, AN2, AN3, and AN4.", "\nTypically, received power in a receiver side can be expressed as:\n P rx = P TX · G TX · G RX ⁡ ( r 4 ⁢ ⁢ π ⁢ ⁢ λ ) 2 · e - α ⁢ ⁢ r where PTX is transmitted power, GTX and GRX are beamforming gains of transmitting and receiving antennas, respectively, λ is the wavelength, and α is the attenuation factor due to absorption in the medium. ", "For an MMW-wave link at 60 GHz, oxygen absorption loss can be as high as 16 dB/km.", "\nFrom the above formula, it is clear that the attenuation of radio wave is proportional to 1/λ2. ", "With the same propagation distance, 60 GHz attenuates 29.5 dB more compared to 2 GHz, without considering the oxygen absorption.", "\nIn considering this, high gain beam-forming is mandatory in order to compensate the extra attenuation. ", "Thanks to the small wavelength, more antenna elements can be integrated in the antenna panel with the same size. ", "This makes it possible to reach a higher beam-forming gain. ", "However, if there are several tens or several hundreds of antenna elements, one Radio Frequency (RF) chain (either TX RF chain or RX RF chain) for each antenna element is inapplicable due to unacceptable cost. ", "In such a case, multiple antenna elements share one RF chain and a specific analog phase adjustment is applied for each antenna in order to adjust the beam direction and maximize the beam-forming gain. ", "Due to the narrow TX beam, it is needed to steer transmission of beacon signals to enable AN discovery area, and to preform beam-forming training to maximize the beam-forming gain.", "\nMeanwhile, high gain beam-forming could bring challenges, including, e.g., hidden problem and deafness problem.", "\nFIG. ", "2 illustrates an example of the hidden problem caused by directivity of high gain beam-forming. ", "As shown in FIG. ", "2, link pair 1 is composed by Access Point 1 (AP1) and User Equipment 1 (UE1), and link pair 2 is composed by AP2 and UE2. ", "When AP2 is transmitting to UE2, neither AP 1 or UE 1 can detect the channel utilized by AP2 and UE2 because both AP1 and UE1 are outside of the TX beam coverage from AP2 to UE2. ", "However, when AP1 transmits data to UE1, its TX beam can reach UE2 and cause interference.", "\nFIG. ", "3 illustrates an example of the deafness problem caused by directivity of high gain beam-forming. ", "As shown in FIG. ", "3, UE 1 and AP1 compose link pair 1 and UE2 and AP2 compose link pair 2. ", "The link pair 2 has ongoing data transmission from AP2 to UE2. ", "But this is not detected by UE1 because UE1 does not monitor (or sense) this direction. ", "However, when UE 1 starts data transmission, the data receiving by UE2 can be clearly impacted due to UE1 and UE2 are close to each other.", "\nCurrently, it is supposed that the total carrier bandwidth of the MMW-RAT can be up to 1 or 2 GHz. ", "This bandwidth can be composed by a number of sub-band carriers of a certain bandwidth, e.g. 100 MHz. ", "By way of example, FIG. ", "4 illustrates one MMW-RAT carrier with 4 sub-bands. ", "The smallest resource grid in FIG. ", "4 corresponds to a sub-band in the frequency domain and to a subframe in the time domain, and may be referred to as a sounding and sensing resource element. ", "Of course, the sounding and sensing resource element may be also in terms of code.", "\nTo allocate the available resources, a contention based resource allocation scheme and/or a scheduling based resource allocation scheme may be applied in MMW-RAT as the basic policy of collision avoidance. ", "A contention based resource allocation scheme provides a mechanism to compete for the channel based on the self-determination on the channel availability. ", "In a scheduling based resource allocation scheme, a scheduler, e.g., a CCU as shown in FIG. ", "1, gains the resource controllability first via either contention based method or coordination method first and allocates the resource to controlled links.", "\nThere could be certain combination of the contention based resource allocation scheme and the scheduling based resource allocation scheme. ", "FIG. ", "5 illustrates an example of a complex interference situation in a MMW-RAT network. ", "As shown in FIG. ", "5, due to directivity of high gain beam-forming, Link 1 and Link 2 may have unendurable UpLink (UL) to DownLink (DL) interference while Link 5 and Link 6 may have unendurable DL to DL interference and UL to DL interference.", "\nDue to directivity of high gain beam-forming, the collision determination is more complex than omni-transmission. ", "The traditional measurement does not work well due to the aforementioned deafness and hidden problems. ", "Besides, though carrier sensing methods commercially used in Wireless Local Area Network (WLAN, 802.11) and Wireless Personal Area Network (WPAN, 802.15) are developed, they are mainly for local access system. ", "It is a distributed carrier sensing scheme, i.e., the carrier sensing is done by each node pair independently. ", "For MMW RAT, firstly it is expected that there can be better dimensioned deployment involving multiple nodes of APs and UEs, and better network controllability (e.g., self-optimization, self-organization, and mobility) than Wireless Fidelity (WiFi) is targeted. ", "Secondly, MMW RAT is expected to provide much better Quality of Service (QoS) than WiFi. ", "In this sense, a better measurement than simple distributed carrier sensing of WiFi is desired.", "\nThe interference measurements in 3G and 4G wireless systems are mainly designed to measure the inter-cell/inter-Transmission-Point interference, rather than inter-link interference. ", "Due to small sector size and the large overlapping coverage in case of MMW RAT, the similar measurement as 3G or 4G systems is not enough to identify links in collision and help the interference management." ]

{ "pile_set_name": "USPTO Backgrounds" }

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[ "Free The Wedding Ringer in High Quality Video with movie summary \"Doug Harris is a loveable but socially awkward groom-to-be with a problem: he has no best man. ", "With less than two weeks to go until he marries the girl of his dreams, Doug is referred to Jimmy Callahan, owner and CEO of Best Man, Inc., a company that provides flattering best men for socially challenged guys in need. ", "What ensues is a hilarious wedding charade as they try to pull off the big con, and an unexpected budding bromance between Doug and his fake best man Jimmy.\" ", "in best look. ", "Download full The Wedding Ringer in High Quality by visiting the download link.", "\n\nNow you can see The Wedding Ringer in HD quality. ", "play full The Wedding Ringer video with single prepare the variety by exploring the image in the sign in place button. ", "Enjoy each of the albums completely new movie using all preferred movie buffering! ", "just play this movie The Wedding Ringer this time for full and free this online movie. ", "That is truly spectacular and may a type of rare remarkable. ", "The good quality was great all around. ", "Management, graphics and aesthetic effects were being all extremely innovative along with brilliant. ", "The pleasure of the script, often humorous and possesses lots of heart for many his characters are extremely very well toned.", "\n\nSure, now you can view movie regarding The Wedding Ringer 100 % length and find the link to this video The Wedding Ringer in high quality.", "\n\nTags: impersonator, wedding,\n\nYou've just seen the movie categories impersonator\ntitled The Wedding Ringer (2015). ", "You can bookmark this page with the URL http://meinkleinerideenkasten.blogspot.de/2015/10/the-wedding-ringer-2015.html. ", "Thank you!" ]

{ "pile_set_name": "Pile-CC" }

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[ "Field\nExemplary embodiments relate to a gate driving circuit. ", "More particularly, the present disclosure relates to a gate driving circuit directly integrated on a display panel.", "\nDiscussion of the Background\nA display device typically includes gate lines, data lines, and pixels. ", "Each of the pixels may be connected to a corresponding gate line of the gate lines and a corresponding data line of the data lines. ", "The display device typically includes a gate driving circuit to control the gate lines and a data driving circuit to control the data lines. ", "The gate driving circuit may apply gate signals to the gate lines, respectively, and a data driving circuit may apply data signals to the data lines, respectively.", "\nThe gate driving circuit typically includes a shift register including driving stage circuits, e.g., driving stages. ", "Each driving stage may output the gate signal corresponding to the gate line. ", "Each driving stage may include transistors connected to each other.", "\nThe above information disclosed in this Background section is only for enhancement of understanding of the background of the inventive concept, and, therefore, it may contain information that does not form the prior art that is already known in this country to a person of ordinary skill in the art." ]

{ "pile_set_name": "USPTO Backgrounds" }

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[ "oklahomadawn says:\nI bet Cooper loved all the attention!!! ", "outstanding extraction and such an adorable spring scene!! ", "Cooper's bunny ears are so cute and your finishing edits beautiful!! ", "thanks so much for posting your scrap in This Week's Favorites group!! ", "Great Favorite!!!Apr 8, 2013" ]

{ "pile_set_name": "Pile-CC" }

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[ "Ryusei Saito\n\nis a Japanese footballer who plays for FC Osaka.", "\n\nClub statistics\nUpdated to 23 February 2019.", "\n\nReferences\n\nExternal links\n\nProfile at Kyoto Sanga\nProfile at Fujieda MYFC\n\nCategory:1994 births\nCategory:Living people\nCategory:Association football people from Osaka Prefecture\nCategory:Japanese footballers\nCategory:J2 League players\nCategory:J3 League players\nCategory:Japan Football League players\nCategory:Kyoto Sanga FC players\nCategory:SP Kyoto FC players\nCategory:FC Osaka players\nCategory:Fujieda MYFC players\nCategory:Association football defenders\nCategory:People from Tondabayashi, Osaka" ]

{ "pile_set_name": "Wikipedia (en)" }

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[ "Mom Answers\n\nWas your Dr. refering to your baby or you? ", "Just wondering. ", "I don't understand what Dr.'s mean when they say this. ", "I'm 25 weeks & at my last visit, my Dr. told me the baby was measuring a few ounces over than average & I was under-weight.", "\n\nThis Internet site provides information of a general nature and is designed for educational purposes only. ", "If you have any concerns about your own health or the health of your child, you should always consult with a physician or other healthcare professional. ", "Please review the Terms of Use before using this site. ", "Your use of the site indicates your agreement to be bound by the Terms of Use.", "\n\nThis site is published by BabyCenter, L.L.C., which is responsible for its contents as further described and qualified in the Terms of Use." ]

{ "pile_set_name": "Pile-CC" }

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[ "Cooling mechanisms are commonly implemented to constrain the temperature of high-heat-flux devices within an allowable range. ", "Thermal drift can result in various negative consequences, including decreased efficiency, reduced life, and even device failure. ", "When a high-power device actuates in a transient manner, the challenge of maintaining stable temperature is considerably more difficult, especially if the thermal time constant of the device is relatively small. ", "Indeed, the cooling must be cycled very rapidly to match the pulsed heat load.", "\nFor example, laser diode arrays require uniform temperature, both spatially and temporally, to reduce wavelength shift and spectral broadening. ", "A lasing event is commonly implemented as a series of high current density quasi-continuous-wave pulses that are between 10 μs to 1 ms in duration and induce a local temperature change of 1-10° C. This short timescale thermal behavior is overlaid on a longer timescale response resulting from the average power (˜100 ms). ", "The spectral output of an 808 nm GaAs quantum-well semiconductor laser shifts by 0.27 nm ° C.−1 as the mean temperature of the device drifts.", "\nIn this and similar applications, the objective is to keep the device at a near-constant temperature throughout the duration of device actuation. ", "To achieve this, the cooling mechanism must be tuned to the thermal mass and heat output of the device. ", "A higher initial rate of cooling would offset the thermal mass of the device, preventing thermal overshoot during the transient start-up period, while ongoing cooling could ensure stable temperature after the initial thermal transients die out.", "\nUnder such conditions, a cooling mechanism designed for transient performance can significantly improve device performance. ", "Of the commonly employed cooling approaches capable of moderate-to-high heat flux, some are better suited to transient actuation than others. ", "While pool/flow-boiling processes typically have long start-up periods that are not ideal, other methods such as jet-impingement, spray, and microchannel cooling are more promising candidates for transient performance. ", "Nevertheless, none of these conventional cooling techniques rely on inherently transient phenomena—indeed, they assume steady state and are only initiated and then sustained continually. ", "With conventional techniques, even if the cooling mechanism can be activated in a stepwise manner, the thermal resistance and capacitance of the system results in thermal fluctuations that cannot be entirely eliminated.", "\nConversely, flash boiling of a liquid pool is unique in that the phenomenon is inherently transient, with phase-change typically initiating at a maximum and decaying with time. ", "In contrast, phase change in many conventional architectures is limited to a thermal boundary layer near the heated surface, characterized by the wall superheat.", "\nThe term “flash boiling” includes a number of different phase-change processes that initiate from a pressure drop, whether due to the fluid flowing through a pressure gradient or the depressurization of an otherwise stationary body of fluid. ", "This rapid depressurization to a condition below the saturation pressure causes the vapor pressure of the liquid to meet or exceed the pressure of its environment and results in a body of liquid that is uniformly superheated. ", "The liquid superheat, which is not easily attainable in traditional boiling hierarchies and is typically characterized by the Jakob number, provides the requisite latent enthalpy for rapid phase change and, as a result, high rates of expansion and cooling.", "\nThe flash boiling of a stream of fluid, i.e., a flashing flow, is commonly used for atomization processes in which a liquid is injected into an environment where the pressure is at or below the vapor pressure of either the fluid or one of its constituents. ", "Typically, upon injection the flashing fluid is in a significantly superheated state, resulting in a rapid phase-change process and an associated fine dispersion. ", "This technique is considered to be one of the more effective methods of atomizing a fluid and its use ranges from domestic aerosols to direct fuel injection.", "\nThe alternate scenario, in which a body of fluid is depressurized, has been studied at widely varying degrees of nonequilibrium ranging from highly restricted venting of pressurized vessels to explosive homogenous nucleation at the superheat limit. ", "As it does with flashing flows, the rate and the magnitude of the pressure drop significantly alters the nature of the phase-change process experienced by the fluid. ", "If the rate of depressurization is sufficiently low, the liquid-gas system can be considered to remain in thermodynamic equilibrium throughout the entire event, and the phase change can be treated as a thermodynamically limited process. ", "More commonly however, a certain degree of superheat will be attained by the liquid as controlled by the kinetics of the phase-change process.", "\nOnce the fluid attains a certain level of superheat, heterogeneous boiling may initiate depending on the size and density of nucleation sites available and, in transient processes, the amount of time the fluid dwells at the given level of superheat. ", "Higher levels of superheat will typically result in a faster heterogeneous boiling process as more nucleations sites become active. ", "These nucleation sites can include surface defects that are not fully wetted and absorbed gas within the liquid. ", "A liquid can remain in this metastable state for a practically indefinite amount of time if no sufficient sites are available.", "\nIf the superheat is attained quickly enough as determined by the nucleation sites available, the fluid can reach the superheat limit and initiate homogeneous boiling. ", "At this limit, the fluid reaches a point of thermodynamic instability, and boiling occurs volumetrically throughout the fluid irrespective of the presence of nucleation sites.", "\nFlash boiling of a pool of liquid can also occur at intermediate levels of superheat. ", "Upon depressurization, the phase-change process commonly is manifested as a boiling front that propagates from the liquid-gas interface downwards through the liquid. ", "Alternately, the boiling event can initiate and propagate from the surfaces of the vessel.", "\nDespite the finite duration of the event, flash-boiling may still be suitable for transient applications that require pulsed cooling for time periods in the range of 100 milliseconds to 10 seconds. ", "In particular, the flash results in an initial peak cooling that wanes to a quasi-steady state value, which may be useful for counteracting the thermal mass of a heated device to achieve a more constant transient temperature. ", "What is needed are heat management methods that apply flash boiling to heat-heat-flux cooling, as well as convective cooling systems that utilize flash boiling—or similar transient phenomena—to efficiently transfer heat from a hot body via a phase change and/or conduction." ]

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[ "<?", "php\n\nnamespace App\\Containers\\Localization\\Tests\\Unit;\n\nuse App\\Containers\\Localization\\Tasks\\GetAllLocalizationsTask;\nuse App\\Containers\\Localization\\Tests\\TestCase;\nuse App\\Containers\\Localization\\Values\\Localization;\nuse Illuminate\\Support\\Facades\\App;\nuse Illuminate\\Support\\Facades\\Config;\n\n/**\n * Class GetLocalizationsTest.", "\n *\n * @group localization\n * @group unit\n */\nclass GetLocalizationsTest extends TestCase\n{\n\n /**\n * @test\n */\n public function test_if_all_supported_languages_are_returned()\n {\n $class = App::make(GetAllLocalizationsTask::class);\n $localizations = $class->run();\n\n $configuredLocalizations = Config::get('localization-container.supported_languages', []);\n\n // assert that they have the same amount of fields\n $this->assertEquals(count($configuredLocalizations), $localizations->count());\n\n // now we check all localizations in particular\n }\n\n public function test_if_specific_locale_is_returned()\n {\n $class = App::make(GetAllLocalizationsTask::class);\n $localizations = $class->run();\n\n $unsupportedLocale = new Localization('fr');\n\n $this->assertContainsEquals($unsupportedLocale, $localizations);\n }\n\n public function test_if_specific_locale_with_regions_is_returned()\n {\n $class = App::make(GetAllLocalizationsTask::class);\n $localizations = $class->run();\n\n $unsupportedLocale = new Localization('en', ['en-GB', 'en-US']);\n\n $this->assertContainsEquals($unsupportedLocale, $localizations);\n }\n\n public function test_if_wrong_locale_is_not_returned()\n {\n $class = App::make(GetAllLocalizationsTask::class);\n $localizations = $class->run();\n\n $unsupportedLocale = new Localization('xxx');\n\n $this->assertNotContainsEquals($unsupportedLocale, $localizations);\n }\n}\n" ]

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[ "Na caótica e populosa capital indonésia, Jacarta, caminhar nas ruas à noite não é uma atividade de risco ou de ousadia — Foto: Fuzz/Creative Commons\n\nNa caótica e populosa capital indonésia, Jacarta, caminhar nas ruas à noite não é considerada uma atividade de risco ou de ousadia. ", "A Indonésia - o quarto país mais populoso do mundo - é um dos lugares com menor índice de violência no Sudeste Asiático, superado apenas pela rica cidade-Estado Singapura.", "\n\nOs desafios da realidade indonésia - que abriga 260 milhões de pessoas - são similares aos brasileiros. ", "Pobreza, desemprego e desigualdade são marcas do país, de maioria muçulmana. ", "Além disso, há um enfrentamento constante entre o Estado e grupos independentistas e ameaças de células terroristas, desafio distante do cenário brasileiro. ", "Mas a Indonésia está longe de ser apontada como um país inseguro.", "\n\nDe acordo com o Escritório das Nações Unidas sobre Drogas e Crimes (UNODC), a Indonésia tem 0,50 mortes por cada 100 mil habitantes, equivalente a 1.292 mortes em 2016. ", "No Brasil, em 2017 (dado mais recente), o índice foi de 63.880 homicídios - o maior número de mortes da série histórica - equivalente a 30,8 homicídios para cada 100 mil pessoas.", "\n\nO principal fator que garante a segurança do país é o firme controle e a regulamentação para o uso de armas de fogo. ", "Os números revelam que o baixo índice de criminalidade na Indonésia está diretamente relacionado com a posse de armas.", "\n\nDe acordo com o Sistema Nacional de Monitoramento da Violência (NVMS nas siglas em inglês), entre 2008 e 2013 apenas 6% dos homicídios cometidos por civis na Indonésia envolveram armas de fogo. ", "Este índice inclui armas legalizadas ou não.", "\n\n\"O baixo índice de posse de armas de fogo faz absoluta diferença para inibir assassinatos. ", "Sem dúvida\", afirma à BBC News Brasil Sana Jeffrey, pesquisadora da Universidade de Chicago, que liderou a organização do sistema de monitoramento da violência na Indonésia, desenvolvido pelo Banco Mundial.", "\n\nPara cada arma na Indonésia, circulam 214 no Brasil. ", "Em 2018, o número total de armas de fogo em mãos de civis indonésios era de 82 mil, uma média de 0,3 arma por cada 100 habitantes, de acordo com a ONG Small Arms Survey. ", "No Brasil, o número estimado é de 17,5 milhões de armas, uma média de 8,29 armas para cada 100 pessoas.", "\n\n\"A porcentagem (de crimes) é realmente baixa e a maioria é concentrada em áreas onde há um histórico da guerra civil\", acrescentou Jeffrey.", "\n\nControle de armas\n\nO controle da circulação de armas é um dos elementos prioritários para a polícia local que desde a redemocratização do país, em 1998, luta para se projetar como uma instituição confiável. ", "Para as autoridades, a arma é um sinal de alerta e de associação com organizações criminosas. \"", "É uma questão de manter ou perder o controle e eles são muito sérios nesse aspecto em comparação com outros tipos de crimes\", afirma Jeffey, da Universidade de Chicago.", "\n\nA posse de armas na Indonésia é estrita, porém, permitida. ", "Não há ali um culto às armas como acontece em outros países. ", "Para obter a licença é preciso ter mais de 21 anos e apresentar às autoridades uma \"genuína razão\", como caça ou autodefesa para portar uma arma. ", "Avaliação psicológica e ficha criminal limpa são requisitos obrigatórios. ", "O candidato deve se submeter a um treinamento teórico e prático sobre o uso da arma. ", "Não há lojas de armamentos. ", "A polícia é quem se responsabiliza pela venda direta tanto da arma de fogo como das munições. ", "A licença deve ser renovada a cada dois anos.", "\n\nApesar do controle estatal, o mercado ilegal de armas também é uma realidade no país e corresponde à metade das armas que circulam entre os civis. ", "O armamento irregular é contrabandeado dos países vizinhos ou elaborado artesanalmente.", "\n\nPolícia comunitária e vigilantismo\n\nOutra peça para entender o quebra-cabeça da segurança indonésia são as associações de bairros chamadas de Rukun Tetangga, em bahasa, ou comunidades harmoniosas na tradução literal. ", "Essas associações organizam um sistema de patrulhamento na vizinhança durante a noite. ", "Além dos moradores, há postos policiais que vigiam os bairros e atuam de forma coordenada com os civis.", "\n\nAs Rukun Tetangga foram impostas durante a colonização japonesa (1942-1945). ", "Esse sistema de organização, que pode chegar a funcionar como uma espécie de serviço de inteligência comunitário - se manteve como um instrumento do Estado indonésio para vigilância e controle.", "\n\n\"Todos estão de olho em todos. ", "E apesar de assumirem características autônomas e de autodeterminação, na prática são estruturas extremadamente controladas (pelo Estado)\", afirma à BBC News Brasil a pesquisadora Jacqui Backer, professora de Política do Sudeste Asiático na Universidade de Murdoch, na Austrália.", "\n\nBacker afirma que o Estado exerce um forte controle sobre como as pessoas interagem e vivem suas vidas. \"", "É um Estado vigilante e incrivelmente policiado. ", "Isso pode trazer benefícios em termos de controle de taxas de criminalidade, mas também tem efeitos negativos\", afirma.", "\n\nNa visão de Backer, um dos aspectos perigosos dessa lógica de vigilância comunitária é o poder de instrumentalização que o Estado - religioso e conservador - possui para determinar comportamentos e punir as minorias. \"", "Se você mora em um bairro que tem esse sistema de organização e você tem políticos que estão falando mal sobre gays ou sobre o cristianismo, haverá um efeito negativo direto nesses moradores que representam as minorias\", afirma.", "\n\nEsse controle social é uma das bases do crescente vigilantismo - um dos fenômenos sociais que têm preocupado cada vez mais a acadêmicos e organizações de direitos humanos.", "\n\nVigilantismo ocorre quando um indivíduo ou um grupo de pessoas atuam extrajudicialmente, argumentando fazer \"justiça\" por conta própria. ", "A mira desses grupos varia entre ajuste de contas contra assaltantes à perseguição contra homossexuais, minorias religiosas e suspeitos de praticar feitiçaria. \" ", "O crime (comum) não é um problema na Indonésia, mas o vigilantismo sim é\", afirma Jeffrey.", "\n\nAs vítimas podem sofrer agressões verbais e linchamentos - que em alguns casos terminam em assassinatos. ", "De acordo com o Banco Mundial, entre 2005 e 2014, foram registrados 34 mil casos de agressões de vigilantes, das quais 1.600 resultaram em assassinatos, de acordo com a mídia local. \"", "A maior parte das agressões é feita por residentes que não são afiliados politicamente a nenhum partido ou organização\", afirma Sana Jeffrey, que desenvolve uma tese sobre o crescimento do vigilantismo.", "\n\nEsse tipo de violência é contraditoriamente mais alto nas ilhas de Java e Sumatra - onde há uma forte presença do Estado e níveis muito mais baixos de distúrbios, confrontos e crimes violentos do que o restante do país. ", "Para a pesquisadora Sana Jeffrey, essas ações também demonstram uma falta de credibilidade do poder de ação da polícia como mediadora de conflitos.", "\n\nA maioria dos crimes cometidos pelos vigilantes permanece impune.", "\n\nNas ilhas dedicadas ao turismo, como Bali, as lideranças comunitárias atuam como informantes para monitorar a movimentação de turistas, cujas irregularidades podem variar entre, por exemplo, vistos de permanência expirados e posse de drogas - que pode ser punida com a pena capital.", "\n\nHistória traumática\n\nEsse país de vigiados e vigilantes tem uma raiz histórica traumática.", "\n\nEm 1965, Hadji Mohamed Suharto liderou um golpe de Estado dando início a uma ditadura sangrenta por mais de 30 anos. ", "Entre 1 milhão e 3 milhões de pessoas foram perseguidas e executadas. ", "As vítimas eram democratas, comunistas e outros cidadãos não alinhados ao regime. ", "O massacre na Indonésia ocorreu em meio à Guerra Fria e era de conhecimento dos Estados Unidos, que à época disputavam a influência ideológica na região.", "\n\nO massacre ainda é uma ferida aberta. ", "Uma das maneiras de sobreviver à violência era demonstrar lealdade à Suharto, denunciando supostos comunistas. \"", "A violência ocorreu na base da sociedade, foi uma violência muito particular que fez com que todos fossem cúmplices\", afirma Jacqui Backer. ", "A seu ver, a manutenção da ordem por parte da população também é reflexo do medo." ]

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[ "Mercy Hospital South\n\nMercy Hospital South (formerly St. Anthony's Medical Center until October 1, 2018) is the third-largest medical center in Greater St. Louis and an affiliate of Mercy. ", "It is the only designated Level II Trauma Center in either South St. Louis County or Jefferson County. ", "The hospital is located in unincorporated south St. Louis County, Missouri just south of I-270 along Tesson Ferry Road (Missouri Route 21). ", "It serves families in St. Louis County, St. Louis City, and Jefferson County, as well as Franklin County, St. Francois County, Ste. ", "Genevieve County, and Washington County in Missouri, along with Monroe, Randolph and St. Clair counties in Illinois. ", "In addition, Mercy Hospital South operates four urgent care facilities located in Arnold, Fenton, Kirkwood, and Lemay.", "\n\nMercy Hospital South has 767 licensed beds and is staffed by approximately 3,700 employees, 790 physicians and provides care to approximately 200,000 patients each year.", "\n\nHistory\nIn 1900, the Franciscan Sisters, Daughters of the Sacred Hearts of Jesus and Mary, based in Wheaton, Illinois, opened St. Anthony's Hospital at Grand and Chippewa in south St. Louis City. ", "The facility was named after Anthony of Padua, an early disciple of Francis of Assisi, who patterned his life and healing ministry after the example set by Jesus Christ. ", "\n\nFollowing St. Louis' first polio epidemic in 1946, St. Anthony's Hospital became designated by the National Foundation for Infantile Paralysis (NFIP) as its Midwest Center for polio treatment. ", "In 1947, convalescent children who had survived polio under the auspices of the NFIP—later known as the March of Dimes—and its city and county chapters were brought to this hospital. ", "It became one of the country's largest polio rehabilitation centers, treating more than 100 polio patients per day. ", "In 1954, the incidence of new cases of polio tapered off and the hospital began offering twice-yearly Salk polio vaccine clinics.", "\n\nIn 1967, the Franciscan Sisters transferred ownership and control of the hospital to a board of community leaders, making St. Anthony's the first Catholic hospital in the St. Louis area to be administered by a lay board. ", "In 1975, St. Anthony's Medical Center opened at its present \nlocation at 10010 Kennerly Road. ", "In 2017, St. Anthony's became affiliated with Mercy, which operates Mercy Hospital St. Louis. ", "St. Anthony's officially changed names to Mercy Hospital South on October 1, 2018 to match the Mercy branding.", "\n\nSee Jean Fahey Eberle's history of Oakville-Mehlville-Concord Village Communities.", "\n\nServices\n \nMercy Hospital South offers medical treatment in acute rehabilitation, behavioral health, cardiology/heart care, emergency medicine, hospice, Hospice House (de Greeff),\nneurology/stroke, \noncology/cancer care, \northopedics, \nsurgery, and maternity services.", "\n\nAwards and recognition\n\n In 2014, St. Anthony's Acute Rehabilitation Department received a three-year accreditation from the Commission on Accreditation for Rehabilitation Facilities (CARF).", "\n In 2013, St. Anthony's was the site of the first-in-the-world implantation of a bifurcated stent graft, used to treat patients with aortic and iliac artery aneurisms while preserving blood flow to the pelvis. ", "\n In 2013, St. Anthony's Emergency Department staff was honored with four Abbott EMS \"Bridge to Life\" Clinical Save Awards for its patient care.", "\n In 2013, St. Anthony's Stroke Program was honored with two awards from the American Heart Association and the American Stroke Association for its commitment to quality care.", "\n In 2012, St. Anthony's was one of 49 hospitals nationwide to receive an \"Excellence in Patient Care Award\" and the only hospital honored with the \"Ambulatory Care Unit Award\" from the Studer Group, a healthcare performance improvement and leadership development firm.", "\n In 2011, St. Anthony's Diabetes Education Program received a recognition certificate for quality from the American Diabetes Association.", "\n In 2011, St. Anthony's Medical Center earned the highest level of accreditation available as a Chest Pain Center with PCI (Percutaneous Coronary Intervention) from the Society of Chest Pain Centers. ", "\n In 2011, St. Anthony's earned recertification as a Primary Stroke Center from the Joint Commission on Accreditation of Healthcare Organizations (JCAHO).", "\n In 2009, St. Anthony's received the Hospital Quality Program \"Meritorious Award\" for Improving Patient Throughput, from Anthem Blue Cross and Blue Shield, to recognize its achievements in improving the speed and quality of patient care in its Emergency Department.", "\n In 2008, St. Anthony's earned Quality Respiratory Care Recognition from the American Association for Respiratory Care for the sixth year in a row.", "\n In 2007, St. Anthony's was recognized for patient safety by an independent health ranking firm. ", "\n In 2006, St. Anthony's Medical Center was recognized with a Missouri Team Quality Award. ", "\n In 2005, St. Anthony's became the first hospital in St. Louis to receive Accredited Chest Pain Center designation by the Society of Chest Pain Centers.", "\n In 2005, St. Anthony's was named a Solucient Top 100 performance improvement leader among U.S. hospitals.", "\n In 2004, St. Anthony's was designated one of the \"Best Places to Work\" by the St. Louis Business Journal.", "\n In 2004, St. Anthony's received the Cardiac Center of Excellence designation from United Healthcare, the largest health plan in St. Louis.", "\n In 1999, St. Anthony's received a Top 100 Hospital designation from HCIA for orthopedic surgery – total knee replacement.", "\n\nReferences\n\nExternal links\n Official website \n\nCategory:Healthcare in St. Louis County, Missouri\nCategory:Greater St. Louis\nCategory:Franciscan hospitals\nCategory:Former Catholic hospitals\nCategory:Hospitals established in 1900\nCategory:Catholic hospitals in North America\nCategory:Buildings and structures in St. Louis County, Missouri\nCategory:1900 establishments in Missouri" ]

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[ "Cat Faeries Blog\n\nThoughts and Ideas About The Modern Housecat\n\nCat Treats Recalled – Contamination That Could Cause Anemia\n\nOnce again, food for our animal friends has been contaminated by harmful additives. ", "This time it’s cat treats. ", "Over 1,000 cartons of “Catswell Brand VitaKitty Chicken Breast with Flaxseed and Vitamins” have been recalled by the manufacturer because of contamination by propylene glycol.", "\n\nWhile propylene glycol has been approved by the FDA as an additive for dog and human food, it is banned by the FDA for cat food because it can cause Heinz body anemia, a serious disease for cats that causes the destruction of red blood cells. ", "Symptoms include fever, weakness, loss of appetite, reddish brown urine, skin discoloration and pale lips, mouth, or gums.", "\n\nThis is a continuing and serious problem for cats, dogs and our other animal friends – harmful contaminates in pet food which are introduced somewhere in the supply chain. ", "Often these chemicals are being added overseas, where some pet food companies source their ingredients.", "\n\nSearch the Cat Faeries Blog\n\nNote: We greatly respect everyone's right to privacy, and Cat Faeries will never sell or share your email address or anything about you with anyone. ", "Never, ever! ", "You may unsubscribe anytime. ", "Easy peasy!", "\n\nAbout Cat Faeries\n\nCat Faeries are the feline specialists who will help you get your cat back in the litter box. ", "We are whizzes at harmonizing multi cat households, and keeping cats feeling young and healthy. ", "And cat toys – wowie meowie, our hand made in the US cat toys are legendary! ", "We are always hunting and gathering information to make the lives of cats and the people who love them, better.", "\n\nVisit CatFaeries.com and learn about our wonderful products which we have crafted or sourced for the modern housecat. ", "Easy and secure online shopping awaits you. ", "And with great prices. ", "Be sure to sign up for our newsletter." ]

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[ "Predicate Notice\n\nPredicate notices are notices required by law that must be served on the tenant before commencing an eviction proceeding. ", "The type of notice to be served depends on the specific facts and circumstances of each case. ", "If the landlord does not serve the proper predicate notices before commencing an eviction proceeding, the court may dismiss your case.", "\n\nJames G. Dibbini & Associates, P.C. have over 20 years of experience representing landlords in complex eviction proceedings and are well versed on the strict procedures that must be followed. ", "Our firm can draft the appropriate notices and serve them properly to ensure your case will not be dismissed on procedural grounds. ", "Let us help you. ", "Give us a call at (914) 965-1011 or email us at jdibbini@dibbinilaw.com. ", "In the meantime, click HEREto get started today in resolving your holdover tenant matter." ]

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[ "Get Diapers\n\nWe have partnered with many Valley non-profit agencies that can get you the help you need.", "\n\nEmergency Diaper Need\n\nThe Diaper Bank of Central Arizona provides diapers to partner agencies that work with people in need. ", "Listed below are our partner agencies that have agreed to provide an emergency supplyof diapers to those in immediate need.", "\n\nPlease note: due to our limited supply, these groups are only able to distribute a handful of diapers in a given time period, typically enough to get you through the day or to the next day. ", "Our emergency suppliers cannot provide a regular supply of diapers, as there simply aren’t the resources to allow this.", "If you have ongoing diaper supply needs, please be sure to connect with our partner agencies, who can hopefully work with you on a more comprehensive level to provide support." ]

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[ "Q:\n\ngetting specific data from an object\n\nI'm trying to figure out how to get the data I need out of an object I created.", "\nI previously had my app data stored in 2 arrays like this:\nconst AVATARS = {\n avatar1: require('./images/avatar_1.jpg'),\n avatar2: require('./images/avatar_2.jpg')\n}\n\nexport const STATS = {\n stats1: \"\\nHP: 124\\n\" +\n \"INT: 23\\n\" +\n \"CHR: 13\\n\",\n\n stats2: \"\\nHP: 423\\n\" +\n \"INT: 24\\n\" +\n \"CHR: 11\\n\"\n}\n\nTo populate the app page, I would use these two functions to get the data:\nimport { AVATARS } from './arrays'\nimport { STATS } from './arrays'\n\n//returns data based on array position(num)\nexport function getAvatar(num) {\n return AVATARS['avatar' + num];\n}\n\nexport function getStats(num) {\n return STATS['stats' + num];\n}\n\nI recently decided to switch to an object where I store all the data, like this:\nexport var Characters = [\n {\n id: 1,\n Name: \"Abe\",\n HitPointValue: \"124\",\n StrengthValue: \"12\",\n IntelligenceValue: \"14\",\n WisdomValue: \"16\",\n DexterityValue: \"12\",\n ConstitutionValue: \"10\",\n CharismaValue: \"17\",\n Avatar: require('./images/avatar_7.jpg')\n },\n {\n id: 2,\n Name: \"Jake\",\n HitPointValue: \"141\",\n StrengthValue: \"21\",\n IntelligenceValue: \"6\",\n WisdomValue: \"5\",\n DexterityValue: \"8\",\n ConstitutionValue: \"20\",\n CharismaValue: \"10\",\n Avatar: require('./images/avatar_2.jpg')\n }\n]\n\nBut now that I did that, I have no idea how to get the needed data. ", " Obviously my old functions won't work. ", " Is there a way to pick which data I need for a given page? ", " \nFor example, if I only need the Name and Avatar for one page, but I might need the stats for another page.", "\nThanks!", "\n\nA:\n\nYou can modify you functions to accept id like\nimport {characters} from './arrays';\n\n//returns data based on array position(id)\nexport function getAvatar(id) {\n var idx = characters.findIndex((val) => val.id == id);\n\n return characters[idx].Avatar;\n}\n\nexport function getName(id) {\n var idx = characters.findIndex((val) => val.id == id);\n return characters[idx].Name;\n}\n\n" ]

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[ ".. index:: ! ", "execution, stack, store\n\nConventions\n-----------\n\nWebAssembly code is *executed* when :ref:`instantiating <exec-instantiation>` a module or :ref:`invoking <exec-invocation>` an :ref:`exported <syntax-export>` function on the resulting module :ref:`instance <syntax-moduleinst>`.", "\n\nExecution behavior is defined in terms of an *abstract machine* that models the *program state*.", "\nIt includes a *stack*, which records operand values and control constructs, and an abstract *store* containing global state.", "\n\nFor each instruction, there is a rule that specifies the effect of its execution on the program state.", "\nFurthermore, there are rules describing the instantiation of a module.", "\nAs with :ref:`validation <validation>`, all rules are given in two *equivalent* forms:\n\n1. ", "In *prose*, describing the execution in intuitive form.", "\n2. ", "In *formal notation*, describing the rule in mathematical form. [#", "cite-pldi2017]_\n\n.. note::\n As with validation, the prose and formal rules are equivalent,\n so that understanding of the formal notation is *not* required to read this specification.", "\n The formalism offers a more concise description in notation that is used widely in programming languages semantics and is readily amenable to mathematical proof.", "\n\n\n.. _exec-notation-textual:\n\nProse Notation\n~~~~~~~~~~~~~~\n\nExecution is specified by stylised, step-wise rules for each :ref:`instruction <syntax-instr>` of the :ref:`abstract syntax <syntax>`.", "\nThe following conventions are adopted in stating these rules.", "\n\n* The execution rules implicitly assume a given :ref:`store <store>` :math:`S`.", "\n\n* The execution rules also assume the presence of an implicit :ref:`stack <stack>`\n that is modified by *pushing* or *popping*\n :ref:`values <syntax-value>`, :ref:`labels <syntax-label>`, and :ref:`frames <syntax-frame>`.", "\n\n* Certain rules require the stack to contain at least one frame.", "\n The most recent frame is referred to as the *current* frame.", "\n\n* Both the store and the current frame are mutated by *replacing* some of their components.", "\n Such replacement is assumed to apply globally.", "\n\n* The execution of an instruction may *trap*,\n in which case the entire computation is aborted and no further modifications to the store are performed by it. (", "Other computations can still be initiated afterwards.)", "\n\n* The execution of an instruction may also end in a *jump* to a designated target,\n which defines the next instruction to execute.", "\n\n* Execution can *enter* and *exit* :ref:`instruction sequences <syntax-instr-seq>` that form :ref:`blocks <syntax-instr-control>`.", "\n\n* :ref:`Instruction sequences <syntax-instr-seq>` are implicitly executed in order, unless a trap or jump occurs.", "\n\n* In various places the rules contain *assertions* expressing crucial invariants about the program state.", "\n\n\n.. index:: ! ", "reduction rules, configuration, evaluation context\n.. _exec-notation:\n\nFormal Notation\n~~~~~~~~~~~~~~~\n\n.. note::\n This section gives a brief explanation of the notation for specifying execution formally.", "\n For the interested reader, a more thorough introduction can be found in respective text books. [#", "cite-tapl]_\n\nThe formal execution rules use a standard approach for specifying operational semantics, rendering them into *reduction rules*.", "\nEvery rule has the following general form:\n\n.. math::\n \\X{configuration} \\quad\\stepto\\quad \\X{configuration}\n\nA *configuration* is a syntactic description of a program state.", "\nEach rule specifies one *step* of execution.", "\nAs long as there is at most one reduction rule applicable to a given configuration, reduction -- and thereby execution -- is *deterministic*.", "\nWebAssembly has only very few exceptions to this, which are noted explicitly in this specification.", "\n\nFor WebAssembly, a configuration typically is a tuple :math:`(S; F; \\instr^\\ast)` consisting of the current :ref:`store <store>` :math:`S`, the :ref:`call frame <frame>` :math:`F` of the current function, and the sequence of :ref:`instructions <syntax-instr>` that is to be executed.", "\n(A more precise definition is given :ref:`later <syntax-config>`.)", "\n\nTo avoid unnecessary clutter, the store :math:`S` and the frame :math:`F` are omitted from reduction rules that do not touch them.", "\n\nThere is no separate representation of the :ref:`stack <stack>`.", "\nInstead, it is conveniently represented as part of the configuration's instruction sequence.", "\nIn particular, :ref:`values <syntax-val>` are defined to coincide with |CONST| instructions,\nand a sequence of |CONST| instructions can be interpreted as an operand \"stack\" that grows to the right.", "\n\n.. note::\n For example, the :ref:`reduction rule <exec-binop>` for the :math:`\\I32.\\ADD` instruction can be given as follows:\n\n .. math::\n (\\I32.\\CONST~n_1)~(\\I32.\\CONST~n_2)~\\I32.\\ADD \\quad\\stepto\\quad (\\I32.\\CONST~(n_1 + n_2) \\mod 2^{32})\n\n Per this rule, two |CONST| instructions and the |ADD| instruction itself are removed from the instruction stream and replaced with one new |CONST| instruction.", "\n This can be interpreted as popping two value off the stack and pushing the result.", "\n\n When no result is produced, an instruction reduces to the empty sequence:\n\n .. math::\n \\NOP \\quad\\stepto\\quad \\epsilon\n\n:ref:`Labels <label>` and :ref:`frames <frame>` are similarly :ref:`defined <syntax-instr-admin>` to be part of an instruction sequence.", "\n\nThe order of reduction is determined by the definition of an appropriate :ref:`evaluation context <syntax-ctxt-eval>`.", "\n\nReduction *terminates* when no more reduction rules are applicable.", "\n:ref:`Soundness <soundness>` of the WebAssembly :ref:`type system <type-system>` guarantees that this is only the case when the original instruction sequence has either been reduced to a sequence of |CONST| instructions, which can be interpreted as the :ref:`values <syntax-val>` of the resulting operand stack,\nor if a :ref:`trap <syntax-trap>` occurred.", "\n\n.. note::\n For example, the following instruction sequence,\n\n .. math::\n (\\F64.\\CONST~x_1)~(\\F64.\\CONST~x_2)~\\F64.\\NEG~(\\F64.\\CONST~x_3)~\\F64.\\ADD~\\F64.\\MUL\n\n terminates after three steps:\n\n .. math::\n \\begin{array}{ll}\n & (\\F64.\\CONST~x_1)~(\\F64.\\CONST~x_2)~\\F64.\\NEG~(\\F64.\\CONST~x_3)~\\F64.\\ADD~\\F64.\\MUL \\\\\n \\stepto & (\\F64.\\CONST~x_1)~(\\F64.\\CONST~x_4)~(\\F64.\\CONST~x_3)~\\F64.\\ADD~\\F64.\\MUL \\\\\n \\stepto & (\\F64.\\CONST~x_1)~(\\F64.\\CONST~x_5)~\\F64.\\MUL \\\\\n \\stepto & (\\F64.\\CONST~x_6) \\\\\n \\end{array}\n\n where :math:`x_4 = -x_2` and :math:`x_5 = -x_2 + x_3` and :math:`x_6 = x_1 \\cdot (-x_2 + x_3)`.", "\n\n\n.. [#cite-pldi2017]\n The semantics is derived from the following article:\n Andreas Haas, Andreas Rossberg, Derek Schuff, Ben Titzer, Dan Gohman, Luke Wagner, Alon Zakai, JF Bastien, Michael Holman. ", "|PLDI2017|_. ", "Proceedings of the 38th ACM SIGPLAN Conference on Programming Language Design and Implementation (PLDI 2017). ", "ACM 2017.", "\n\n.. [#cite-tapl]\n For example: Benjamin Pierce. ", "|TAPL|_. ", "The MIT Press 2002\n" ]

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[ "Sckoon Menstrual Cup Review\n\nTampons and menstrual pads are the go-to products for most women. ", "They are what you see sold in traditional stores and what you see advertised in TV commercials and magazine ads. ", "They are far from the only options women have though. ", "Hopefully by now you have also heard of some of the alternatives…cloth pads, natural tampons (sponges), and of course menstrual cups.", "\n\nLet’s start off by saying that this product was not my first rodeo with menstrual cups. ", "I have been using them for years. ", "I have tried several different brands and have been happy with all them. ", "Last month though was my first time using the Sckoon cup brand. ", "Before that I used to purchase their cloth diapers and baby clothes…though that was some time ago.", "\n\nA menstrual cup does what a tampon or pad does but rather than absorb the flow it collects it. ", "It IS a cup. ", "If you are not squeamish about inserting a tampon then you should not be squeamish about inserting a cup. ", "It is basically the exact same thing. ", "The only difference is that you pull a tampon out with a string and toss it in the toilet. ", "With a cup you pull it out, empty it into the toilet, and then wash it for re-use. ", "I guess it is slightly more complicated then a pad or tampon but once you get the hang of it, it is no big deal.", "\n\nI like menstrual cups more than I ever liked tampons. ", "For whatever reason I could always feel tampons, even though they say you shouldn’t. ", "With a cup…I truly cannot feel it. ", "Tampon applicators are also really slippery, which meant many times I had problems with insertion and had to use toilet paper to kind of dry it off and try again. ", "With one brand of menstrual cup I have had some launch failures for the same reason, slippery-ness. ", "With the Sckoon cup there are neato grooves on the bottom (of their otherwise silky smooth and comfortable cup) that give you something to grip and hence no issues with that. ", "See them below…\n\nWhile on the subject of how it compares to other cups let me also use this same photo to show you how it has that little stem sticking out of the bottom of the cup. ", "The other brands have that too but instead of a solid stem that tapers off they have a little hollow tube. ", "That tube collects menstrual flow and has to be washed out…this is not an issue with the Sckoon. ", "This may seem like a small detail but I really like it.", "\n\nI also love that the Sckoon cups come in COLORS!!! ", "My other cup is clear and it has started to look dingy over time. ", "I am not willing to use anything harsher than soap and water to clean it because I want it to last. ", "These colored cups solve that issue because they will stay pretty and not get discolored. ", "I have blue obviously but they also come in other colors.", "\n\nAs with other cups it comes in two sizes. ", "Size 1 is for people who have never given birth vaginally. ", "Size 2 is for those of us who have given birth vaginally. ", "It is made from FDA approved soft medical-grade silicon, so it’s safe, reusable, environmentally friendly.", "\n\nIf you need a crash course in how to use a menstrual cup it is really very easy… Take the cup in hand and fold it using your fingers. ", "Insert pushing up and backwards (towards your back). ", "Your vaginal muscles will grip it and keep it in place and it will open once it is up there too. ", "When you are ready to take it out simply bear down a little bit and the bottom part will come out. ", "Grip it and pull it out carefully so you don’t spill. ", "Dump the contents, wash it with soap and water, and re-insert.", "\n\nThe only time when this becomes a bit tricky is when you are using a public bathroom. ", "I simply dump it out and reinsert it and then remember to wash it as soon as I get home. ", "I also use toilet paper from the bathroom stall to clean my hands off so I don’t leave a public bathroom stall with blood all over them. ", "No need to freak anyone out. ", "I don’t imagine that it is any harder than inserting a tampon in a public place.", "\n\nAfter using it through two menstrual cycles I will be reaching for my Sckoon cup before my “other” cup now. ", "It gets an A++ in my book!", "\n\nConnect With Me…\n\nMeet Tiffany\n\nMy name is Tiffany and I am the blogger behind Naturemoms. ", "I live on an urban homestead in Ohio with my husband, three children, and assorted furry friends. ", "When I am not blogging I am usually thrift store shopping, gardening, wildcrafting and food foraging, or otherwise enjoying nature. ", "Enjoy! ", "Read More…" ]

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[ "Intellectual whiplash! ", "The tea party, of course, is defined by its skepticism of big government, and particularly by its dislike of taxes. ", "Puig not only went to work for state government, she represents the people who collect the taxes to keep the beast purring along." ]

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[ "Verkko-ostaja voi saada huolestuttavan lupakyselyn eteensä. ", "Kyse on Klarnan uudesta maksupalvelusta, mutta miksi tietoja ylipäätään pyydetään?", "\n\nRedditissä ihmetellään eräässä suomalaisessa verkkokaupassa eteen tullutta lupapyyntöä maksun yhteydessä. ", "Nordean tunnistautuminen ohjasi maksupalveluyhtiö Klarnan lupakyselyyn, jossa pyydettiin pääsyä ostajan tilitietoihin. ", "Niihin lukeutuivat muun muassa tilinumero ja valuutta, tilin saldo, tilin tyyppi ja yksityiskohtainen tapahtumahistoria kahden kuukauden ajalta. ", "Tietoihin pyydettiin pääsyä 90 päiväksi.", "\n\nKun verkkokauppa käyttää verkkopankkia Klarnan kautta, pitää lupapyyntöön suostua. ", "Laaja tietojen kysely poiki paljon yhteydenottoja Klarnalle.­\n\nKyseessä oli suora verkkopankkimaksu, ei Klarnan tarjoama lasku tai osamaksu. ", "Jotkut kokivat tällaisen lupakyselyn epäilyttäväksi ja turhaksi, sillä moni ei halua luovuttaa tilinsä tietoja uudelle ulkopuoliselle taholle.", "\n\nKlarnan maajohtajan Jani Tyynin mukaan kyselyn syynä on Klarnan uusi maksupalvelu, jota verkkokaupat voivat tarjota kuluttajille. ", "Klarnan pitää puolestaan noudattaa Euroopan unionin uutta psd2-maksupalveludirektiiviä, mikä pakottaa avoimuuteen ja sen myötä esitettävään lupakyselyyn. ", "Avoimuuteen velvoittaa gdpr-tietosuoja-asetus.", "\n\nJani Tyyni on Klarnan maajohtaja.­\n\n– Tarjotessaan verkkopankkimaksua Klarna (ja sen tytäryhtiö Sofort) käsittelevät lupakyselyn tietoja vain tehdäkseen maksun, ja tietoja säilytetään vain maksunpalautuksia ja asiakaspalvelua varten, Tyyni sanoo.", "\n\nAlkuperäinen lupakysely osoittautui kuitenkin kuluttajapalautteen myötä liian laajaksi. ", "Nyt sitä on rajattu niin, että enää pyydetään lupaa tilinumeroon ja joissakin tapauksissa tilin valuuttaan ja saldoon. ", "Toisaalta joissakin tapauksissa Klarna ei pyydä mitään tietoja. ", "Siihen vaikuttaa kulloisenkin pankin käyttämä tekniikka. ", "Luvan voimassaoloa on lyhennetty yhteen päivään.", "\n\n– Olemme muuttaneet pyyntöä siitä, mikä se oli Redditin keskustelussa viitatussa näkymässä, joka oli siis vanha prosessi, Tyyni toteaa.", "\n\nPankkien ohi\n\nKlarnan maksutapoja tarjoaa tällä hetkellä noin 6000 verkkokauppaa Suomessa. ", "Näistä suuressa osassa on tämä uusi verkkopankkimaksu yhtenä vaihtoehtona maksutavaksi kuluttajalle.", "\n\nKlarna perustelee uutta palveluaan helppoudella. ", "Siinä ohitetaan pankkien omat tunnistautumisruudut ostamisen nopeuttamiseksi. ", "Kaikki eivät kuitenkaan halua luovuttaa pääsyä pankkitilin tietoihin millekään muulle taholle kuin pankille.", "\n\nLupakysely voi tulla lisääntyvästi eteen verkkokaupoissa sitä mukaa, kun ne ottavat Klarnan palvelun käyttöön. ", "Ainoana vaihtoehtona on vaihtaa maksutapaa esimerkiksi luottokorttimaksuun. ", "Jos verkkokauppa ei anna muuta maksutapaa, lupapyynnöstä kieltäytyminen tarkoittaa ostoksen tekemättä jättämistä.", "\n\n– Jokaisella markkinalla on oma, uniikki kuluttajakäyttäytyminen, ja se mikä toimii muualla, ei aina toimi täällä Suomessa. ", "Tämän palautteen johdosta olemme muuttaneet tunnistautumisen yhteydessä pyydettäviä tietoja ja valtuutuksen kestoa.", "\n\nJohtajan mukaan pyydettävät tiedot riippuvat osittain pankin omasta tekniikasta. ", "Katetietoa on tarvittu maksutoimeksiannon tekemiseksi. ", "Maksuhistoria puolestaan oli osa petostarkastusta.", "\n\n– Teemme edelleen petostarkistuksia, mutta olemme tulleet siihen tulokseen, että emme välttämättä tarvitse tapahtumahistoriaa siihen tarkoitukseen Suomessa.", "\n\nKilpaileeko Klarna asiakkaiden tiedoista?", "\n\nPSD2:n mukaan välttämättömiä kerättäviä tietoja ovat iban-tilinumero, asiakkaan henkilöllisyys ja tilin valuutta.", "\n\n– Olemme saaneet palautetta, että kaikki eivät täysin koe tuntevansa palvelua ja sen toimintaa, ja sitä yritämme nyt parantaa, Tyyni sanoo arvioiden, että palvelua ei selvästikään ole selitetty kuluttajille tarpeeksi hyvin.", "\n\nKlarnan uusittu lupakysely on aiempaa vähemmän utelias.­\n\nMaajohtaja kiistää oletuksen, että Klarnan maksupalvelun tarkoituksena olisi haastaa pankit henkilötietojen käsittelyssä nyt, kun gdpr-asetus antaa kuluttajille oikeuden siirtää tietojaan palvelusta toiseen. ", "Yritykset saattavat kilpailla niistä, koska tieto on viime kädessä valtaa.", "\n\n– Ei, maksupalvelussamme ei ole kyse kuluttajan henkilötietojen siirtämisestä pankeilta. ", "Pyydämme vain välttämättömiä tietoja, jotka tarvitsemme maksupalvelun tarjoamiseksi kuluttajalle. ", "Emme esimerkiksi koskaan säilytä asiakkaiden luottamuksellisia kirjautumistietoja ja vahvistuskoodeja.", "\n\nNäihin tietoihin ei ole Klarnan työntekijöillä tai ulkopuolisilla tahoilla mitään pääsyä, Tyyni vakuuttaa." ]

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[ "Q:\n\nLess brightness after plugging once my macbook pro to a thunderbolt display\n\nI own a Macbook pro (15'4) with Yosemite.", "\nI used to use it without an external display. ", "\nThe brightness was very powerful :) \nI've just plugged a thunderbolt display (Mac 27''), and then removed it. ", "\nI even restarted my MacBook pro right after.", "\nNow, I found that the brightness is not as powerful as before, even on MAX :(\nMy color profile is the one by default (LCD color).", "\nIs there an explanation? ", "\nHave someone ever experienced this?", "\n\nA:\n\nYou could try a NVRAM reset by starting your mac and holding the following keys: \nCMD + ALT + R + P.\n(More info)\nWhen your Mac restarts, you could release the keys.", "\n\nSome say that you have to do this 3 times for the best result, but I can't explain why\n\nMore information about a PRAM/NVRAM reset can be found on the Apple Support Website\nWhen you use your Mac and connect it to displays or change the screen's brightness, it could be that your settings become a bit corrupted. ", "You could erase those settings by preforming a PRAM reset as explained above. ", "\nI can't find any known bugs on the Apple Diagnostics website, you could report it by sending a mail to feedback@apple.com\n\n" ]

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[ "Full-Service Buying Office for the Promotional Products Industry\n\nODM Buying Office in China – new website.", "\n\nODM is pleased to announce that we are launching a new website to update partners on all the specifics of setting up operations in China. ", "Laws change regularly and are quite dependant on specific industry, China province, and type of structure choosen.", "\n\nApart from assisting companies to set up their own operations, ODM is currently offering clients 2 simple solutions for closer cooperation (both with minimum set up costs/work).", "\n\nemploy a dedicated merchandising staff from our team to work for you on full time on retainer." ]

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[ "Philomastiginae\n\nPhilomastiginae is a subfamily of sawflies in the family Pergidae. ", "There are at least four genera and six described species in Philomastiginae.", "\n\nGenera\nThese four genera belong to the subfamily Philomastiginae:\n Cerospastus Konow, 1899\n Ecopatus Smith, 1990\n Philomastix Froggatt, 1890\n Philoperra Smith, 1995\n\nReferences\n\nCategory:Tenthredinoidea" ]

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[ "Nektan is one of the world’s biggest providers of gaming solutions to some of the world’s biggest online casinos. ", "One of the newest players in the gaming industry, in little under five years Nektan have quickly established themselves as a premium provider of games for online pokies, poker, roulette and a host of other games. ", "Known for providing a supreme experience, their games a characterised by high octane thrills, engaging and charming settings, HD graphics and more. ", "Discover our selection of the top online Nektan games for pokies in New Zealand below." ]

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[ "Sarnoff A. Mednick (1928-2015).", "\nPresents the obituary of Sarnoff A. Mednick (1928 -2015). ", "Sarnoff A. Mednick was considered among the most important figures in psychopathology research in his generation. ", "He pioneered the high-risk research design and made numerous contributions to our understanding of creativity and of the origins of schizophrenia and criminality. ", "The son of Jewish immigrants, Mednick was born on January 27, 1928, and was raised in the Bronx in New York City, New York. ", "He earned a bachelor's degree from the City College of New York in 1948 and a master's degree from Columbia University. ", "In 1954, he earned his doctorate in psychology from Northwestern University. ", "Following a postdoctoral fellowship at the University of Chicago, he was appointed an instructor at Harvard University and then became a visiting assistant research professor at the University of California, Berkeley (1958 -1959). ", "In 1968, he became a professor at the New School for Social Research in New York, where he taught until 1977, when he joined the faculty at the University of Southern California until his retirement in August 2008. ", "Mednick, professor emeritus of psychology at the University of Southern California, died of natural causes on April 10, 2015, in Toledo, Ohio. ", "He was 87." ]

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[ "Expand Your Learning!", "\n\nStudents\n\nBe A Leader\n\nHey. ", "Psst. ", "Yeah you! ", "We know what you're doing. ", "You're looking for educational videos that can either expand on what you were taught in class or that you can use for your own projects. ", "Oh and we do it in a safe environment that your parents and teachers can trust.", "\n\nFind What You Need\n\nNo more digging through millions of videos on YouTube only to find random videos of a twerking cat. ", "Here you can get what you need and get your work done. ", "Now you can get back to searching for the ever elusive four second loop of a granny break dancing!", "\n\nWhat Are You Waiting For?", "\n\nIf you’re ready to get started and jump into the free videos and tools that we provide, then create your free profile. ", "Let us make your education easier and more enjoyable with the TeacherTube experience." ]

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[ "Saturday, September 22, 2012\n\nSigns of Ascension in the Operating Room\n\nThe O.R. is not unlike the military: everything is clean, crisp, organized, and efficient. ", "There is a hierarchy in the Operating Room. ", "As an attending physician, I have higher 'rank' than some, and because of that 'rank' I am given a certain amount of respect for it. ", "Our schedule runs like clockwork, or as much as possible in this aim, given emergencies and other unanticipated events that come our way.", "\n\nAs Reiki Doc, I am a Karuna Reiki Master. ", "Because of this, my access to the Higher Dimensions is open. ", "The opening process has a 'cleanse' that has some very particular symptoms. ", "Support is needed by the student as they navigate their own set of changes they experience as they 'move up' in Reiki.", "\n\nSome of these signs are unmistakable to the experienced Karuna Reiki practitioner, so much so that we fondly call a day with these symptoms 'a Karuna Day'. ", "We change our plans, allow time for this process to happen, ride it out, and know that tomorrow is another day.", "\n\nYesterday in the O.R. I saw evidence of multi-dimensionality at work. ", "I didn't say anything to my coworkers, but now I am to you. ", "And looking back, there have been a few coincidences I had not pieced together leading up to yesterday.", "\n\nThe first one that caught my attention was a patient who did not show up for their surgery. '", "Wasn't it next week?'. ", "This is a classic symptom: the distortion of the perception of Time. ", "At Reiki class, we have had people convinced their Karuna class was on a Sunday, not a Saturday, and slept in. ", "These are ordinary, responsible people. ", "This patient, was educated and also someone who has undergone surgeyr with us before.", "\n\nEveryone else shrugged it off, the surgeon saying it was his scheduler, or the patient misunderstanding the options of today or next week given in the preop visit.", "\n\nBut my NEXT case had a patient. ", "But not technician. ", "Sometimes there is special equipment needed for surgery. ", "For example, a laser, or an orthopedic rep, of a neuro-monitoring technician. ", "Efficiency is the bread-and-butter of these support people. ", "Well, one yesterday thought the surgery was on Saturday, not Friday, and didn't come in. ", "The surgeon was angry, shocked, and disappointed. ", "Who wants to come in on Saturday for an elective case? ", "Fortunately the O.R. was able to scramble, and find someone else.", "\n\nTwice in a row? ", "With two different specialties? ", "That is unusual enough to take notice as a Karuna Master.", "\n\nWhat else? ", "There has recently been an abrupt change in the support service to my specialty. ", "I would like to say more, but let us say a 'lower vibration was removed'. ", "It has been Scrub Tech Week at the hospital. ", "Every day, lunch and dinner has been provided by the reps. ", "Nursing is kind, and always invites me to share. ", "Although it was only pizza and sandwiches, I am thankful to have eaten, and not missed a meal. ", "Yesterday for lunch, I had one piece of Hawaiian pizza, and one piece of cheese, avoiding the 'meat on top of meat on top of meat' variety offered. ", "For dinner, the Charge Nurse (the night one, not the day one I have mentioned), asked if I would like her to save a piece for me. ", "Yes, it was leftover from lunch, not fresh. ", "But she asked what I wanted. ", "I said, Hawaiian. ", "She paused, and said, 'I took the last two pieces. ", "I will go write your name on a napkin and give you one of my pieces.'", "\n\nNobody has ever given me something that was for them to eat in the Operating Room, EVER.", "\nKindness, patience, humility, forbearance, Love, Light, honesty, grace, Peace, Truth, and honestly enjoying each other are signs of Spirit. ", "Of multi-dimensionality. ", "Of Light. ", "We are being bombarded with Light at the present. ", "You may not realize it, but it is true.", "\n\nHere is a quote from a certified negative person in the October issue of Reader's Digest, In the article, 'Forget Happiness': ...in our super-positive society, we have an unspoken zero-tolerance policy for negativity.' ", "Author Augusten Burroughs shares, 'I am not a happy person. ", "There are things that do make me experience joy. ", "But joy is a fleeting emotion, like a very long sneeze. ", "A lot of the time what I feel is interested. ", "Or I feel melancholy. ", "And I also frequently feel tenderness, annoyance, confusion, fear, hopelessness. ", "It doesn't all add up to anything I would call happiness.'", "\n\nMr. Burroughs is dysthymic, he is never at the same baseline as others for 'happiness'. ", "He is not clinically depressed, but he is in a mood right above it and under 'happiness'. ", "Energetically-speaking, he is manifesting his own unpleasant environment, much like energetically farting in his own space and complaining about the smell. ", "He is dwelling on, and reinforcing third dimensional behavior.", "If you saw Gaia Portal yesterday, the higher energies are being anchored and the lower 3-D and 4-D energies are being 'transmuted by Gaia'. ", "People like Mr. Burroughs are going to go someplace else where they can continue their soul development at a dimensional level that is appropriate for him. ", "There are no winners or losers, Salusa said yesterday. ", "Everyone will reach the Higher Dimensions at the time that is right for their soul and its development.", "\nNamaste,\n\nReiki Doc\n\nHere are some links:\nFor Readers' Digest. ", "Right now, the article is on page 128 of the October 2012 issue. ", "It is not 'downloadable' but in time I think the archive or search function will help you find the article. ", "The series immediately follows the 50 Secrets Surgeons article, then an excellent one on Find Happiness on page 126. ", "There is a History of Happiness on page 127, and the quoted article is on page 128-129. ", "The images 'What Happy Looks Like' on page 130 to 139 are a must-see. ", "Buddhist monks, dancers, mermaids, artists, a carefree coal miner, are my favorites.http://www.rd.com/\n(If you want to see a fascinating picture and brief article about Jackie Onassis and Queen Elizabeth, check out p. 140, When Jackie Met the Queen. ", "There are some strained smiles there and if you are up on Cobra's Little Red Pill, you will see right through the smiles and understand the complexity of the 'relationship')\n\nSearch This Blog\n\nTranslate\n\nAbout Me\n\nI am a medical doctor who works in a hospital. ", "I do anesthesia. ", "I am also a Reiki practitioner. ", "While I do anesthesia, I also make this form of Energy Healing available to my patients who wish to have it while their procedure is taking place.", "\nI have quite a following at work. ", "Many patients and coworkers request my services. ", "Recovery room RN's love me. ", "All of my patient report the same thing: they felt no pain. ", "One friend's spouse actually made her take two of her pain pills at home because he absolutely could not believe she felt no pain after gallbladder surgery!" ]

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[ "98 F.3d 1335\nNOTICE: Fourth Circuit Local Rule 36(c) states that citation of unpublished dispositions is disfavored except for establishing res judicata, estoppel, or the law of the case and requires service of copies of cited unpublished dispositions of the Fourth Circuit.", "UNITED STATES of America, Plaintiff-Appellee,v.Michael John ACCETTOLA, Defendant-Appellant.", "\nNo. ", "96-4046.", "\nUnited States Court of Appeals, Fourth Circuit.", "\nSubmitted: Oct. 3, 1996Decided: Oct. 16, 1996\n\nPaul M. McKay, MCKAY & MCKAY, Wheeling, West Virginia, for Appellant.", "\nWilliam D. Wilmoth, United States Attorney, Wheeling, West Virginia, for Appellee.", "\nBefore ERVIN, LUTTIG, and MICHAEL, Circuit Judges.", "\nOPINION\nPER CURIAM:\n\n\n1\nMichael John Accettola appeals his conviction, after a bench trial, for making false statements to a grand jury of the United States in violation of 18 U.S.C.A. § 1623(a) (West Supp.1996) alleging there was insufficient evidence to support the conviction. ", " For the reasons following, we affirm.", "\n\n\n2\nWe must affirm a conviction attacked on the grounds of insufficient evidence if \"after viewing the evidence in the light most favorable to the prosecution, any rational trier of fact could have found the essential elements of the crime beyond a reasonable doubt.\" ", " Jackson v. Virginia, 443 U.S. 307, 319 (1979). ", " The essential elements of 18 U.S.C.A. § 1623(a) are: (1) that the defendant's testimony was under oath before a grand jury; (2) that the testimony given was false in one or more of the respects charged; (3) that the false testimony was knowingly given; and (4) the subject matter of the testimony was \"material\" to the grand jury's investigation. ", " United States v. Friedhaber, 856 F.2d 640, 642 (4th Cir.1988). ", " To be material a \"statement must have 'a natural tendency to influence, or [be] capable of influencing,' \" United States v. Gaudin, --- U.S. ---, 63 U.S.L.W. 4611, 4612 (U.S. June 19, 1995) (No. ", "94-514) (quoting Kungys v. United States, 485 U.S. 759, 770 (1988)), or have \"the natural effect or tendency to impede, influence or dissuade the grand jury from pursuing its investigation.\" ", " Friedhaber, 856 F.2d at 642 (citation omitted).", "\n\n\n3\nIn Accettola's sworn testimony before the grand jury, which concerned charges of distribution of controlled substances and conspiracy or attempt to distribute controlled substances under 21 U.S.C. § 841(a)(1) (1994) and 21 U.S.C.A. § 846 (West Supp.1996), Accettola denied that he had seen Tony Foglio and Rocco Mallicone together at a hotel or that he had seen Mallicone pick up Foglio and a marijuana shipment in a truck. ", " The grand jury testimony directly conflicted with written statements Accettola had previously made to the Federal Bureau of Investigation as part of a plea agreement. ", " The prior statement in which Accettola had agreed to be \"completely forthright and truthful\" was made in the presence of Accettola's defense counsel and reviewed by counsel to ensure accuracy and truthfulness.* ", " Thus, the district court found that, based upon the detailed nature of the prior written statement and the circumstances in the which the statement was given, Accettola knowingly gave false testimony to the grand jury. ", " The court also found that Accettola's false statements were material because they cast light on whether a drug conspiracy to distribute marijuana existed between Mallicone and Foglio. ", " Accordingly, we affirm the conviction because viewing the evidence in the light most favorable to the prosecution, any rational trier of fact could have found Accettola guilty of violating 18 U.S.C.A. § 1623(a) beyond a reasonable doubt. ", " Jackson, 443 U.S. at 319; Friedhaber, 856 F.2d at 642.", "\n\n\n4\nWe dispense with oral argument because the facts and legal contentions are adequately presented in the materials before the court and argument would not aid in the decisional process.", "\n\nAFFIRMED\n\n\n*\n Accettola's attorney read the statement to him verbatim and Accettola signed his name at the bottom of each page. ", " Accettola initialed all changes and corrections to the statement\n\n\n" ]

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[ "Cadmium hyperaccumulation leads to an increase of glutathione rather than phytochelatins in the cadmium hyperaccumulator Sedum alfredii.", "\nSedum alfredii has been reported to be a cadmium (Cd) hyperaccumulator. ", "Phytochelatins (PCs) and other thiol (SH)-containing compounds have been proposed to play an important role in the detoxification and tolerance of some heavy metals, but it is not clear whether PCs are responsible for Cd hyperaccumulation and tolerance in S. alfredii. ", "In this study, two geographically isolated populations of S. alfredii were studied: one population grew on an old Pb/Zn mine site, while the other on a non-mine site. ", "The mine population of this species exhibited a stronger heavy metal tolerance than in the other population. ", "Root-to-shoot transport of Cd was higher in population located at the mine site than at the non-mine site. ", "Considerable amounts of Cd were accumulated in leaves and stems of mine plants, while most Cd was distributed in roots of non-mine plants. ", "Non-protein SH in plant tissues of two populations were further investigated by a HPLC pre-column derivatization system. ", "Upon exposure to Cd, no PCs were detected in all tissues of mine population, while an appreciable amount of glutathione (GSH) was observed in the descending order of stem>root>leaf. ", "The concentrations of GSH consistently increased with the increase of exogenous Cd concentrations and time. ", "On the contrary, Cd exposure strongly induced the production of PCs (mainly PC(2) and PC(3)) and GSH in plant tissues of non-mine population, and the concentrations of GSH showed an initial drop over the duration of 7-d exposure. ", "The present results provided strong evidence that PCs are not involved in Cd transport, hyperaccumulation and tolerance in mine population of S. alfredii." ]

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[ "Taj Mihelich Interview7\n\nPhoto by John Povah\n\nTaj Mihelich is one of the most amazing riders in the history of BMX. ", "Not only does he have the ability to do anything he wants on a bike, he’s also an incredible artist, musician, and one of the nicest people you will ever meet. ", "Earlier this year Taj lit the forums on fire when he became sponsored by Giant, and he just got word that his long-term sponsorship with Etnies is coming to an end. ", "We talked to Taj about the changes in his life and what’s coming next in this exclusive Vital BMX interview. ", "Special thanks to John Povah for the photos.", "\n\nHow did you wind up getting on Etnies in the first place?", "\nI can't really remember how I got on exactly. ", "Rooftop had gotten on a bit before and I think they gave him the green light to pick up a few other guys. ", "Joe RIch got on and then I got on soon after. ", "Airwalk had been talking to me about a signature shoe, and so that was a stipulation of my new contract, that we would start on a signature shoe through Etnies.", "\n\nWhat were some of your proudest moments from all those years?", "\nThose years were really like the prime of my riding career so the good moments are far too many to remember and catalog. ", "I got to travel all over and totally live the BMX dream, you know? ", "I got to design three signature shoes, I got voted favorite rider a few times, I had some great contests, met loads of great people, got to film sections for Etnies Forward and Grounded videos... I don't know... it was all amazing!", "\n\nWhat are some things you were able to do in life that being on Etnies helped with?", "\nWell number one, they have been my main sponsor for years. ", "Providing money, support, and friendship so I could focus on bikes entirely. ", "They also have always trusted me to run the sponsorship my way and encouraged me to follow my own direction. ", "Like, they never \"made\" me do this contest or do some demo. ", "I basically always got to keep riding exactly in the place I wanted it to be, and even though Etnies was supporting me so well I never felt pressured to change who I was. ", "I could pick and choose events and competitions I wanted to attend. ", "They also helped me with the Thunderdome park up in Olympia (that never opened), and they helped Joe and I with some of the initial funding for the T-1 ramp.", "\n\n(Here's a little video of the Thunderdome from 2001. ", "Not crazy riding, but you can get a feel for how fun the ramps were. ", "An earthquake damaged the building and so the park never opened. ", "Such a shame!)", "\n\nWhen/how did you get word that it was coming to an end? ", "Was it a surprise?", "\nThe timing of when I got word was pretty funny. ", "I'd been stressing about buying a new car and after going back and forth for a few weeks I finally decided to go for it. ", "At the dealership signing the dotted line on the last piece of paperwork my phone rang. ", "It was the team manager John Povah and I decided I would call him back after I finished signing. ", "I called him back a few minutes later and it was THE CALL. ", "Oh well.... to some degree I knew it was coming and I wasn't surprised. ", "Poor John Povah, we've been friends for years and making that call was really hard for him, I know. ", "It’s all totally cool, though. ", "Him and I just traveled to Ray's MTB park for fun and it was a blast!", "\n\nYou've seen a lot of sides of the pro BMX life that most people don't know exist. ", "What are some things that would surprise riders about life in the BMX industry?", "\nOh... I don't know if I can say because I don't really know anything else. ", "It’s been my life for as long as I can remember. ", "I think things are a lot more casual then a lot of people realize... for the most part anyway. ", "Or, at least at my level. ", "Some of the big TV contest guys have agents and stuff and that’s a whole other level of business. ", "All of my sponsors have always been really laid back. ", "Sometimes there are legal contracts, but for the most part it’s all just pretty much a “you help us and we'll help you” kind of agreement.", "\n\nYou've had some major sponsorship changes over the past couple of years. ", "Has that affected you're riding at all?", "\nI think everything effects your riding in someway. ", "Riding for Giant has been a big breath of fresh air for me. ", "I've been just a bum on a bike for a while now. ", "No real responsibilities outside of riding and just getting to have a lot of fun. ", "Like seriously, I have been smiling and laughing a lot more lately... bikes are good.", "\n\nFrom skits with Monkey Boy to huge hospital bills, you've been through a lot. ", "Anything you would have done differently over the last stage of your \"career?\"", "\nI don't think so. ", "I've had some memory issues after some concussions so I don't remember a lot of things, so maybe that helps. ", "Only sort of kidding there... at least as far as I can recall everything has been great, or at least worked out in the end in a way that is just fine. ", "Even Monkey Boy... jeez... even that. ", "I'm really excited about the choices I've made lately, too. ", "It’s exposing me to so many fun things and challenging me.", "\n\nWhat's next for you? ", "Austin for life?", "\nWell, Giant said they wanted to try and step things up for me next year so I could remain on my bike. ", "That will be fun if that all works out and I can keep enjoying this full time. ", "I'm thinking a lot about moving from Austin lately. ", "Maybe I just need a change of scenery or something. ", "That’s why I bought a car anyway, to go explore a bit this winter and see if there's anywhere else that feels like it could be home. ", "It’s very likely that my first encounter with snow will send me running to the comfort of warm old Austin.", "\n\nTaj and Etnies have always been a staple. ", "I remember the reason I wanted Etnies when I was younger was because Taj rode them. ", "The first Etnies ad I ever saw was with Taj doing a table I think. ", "Although it sucks he is no longer a part of the team, many will remember the impact on bmx he had while he was on the team." ]

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[ "Q:\n\nIssue with regsub on a string with square brackets with variables in tcl\n\nI am unable to have the current pattern for regex substitution on a string that has square brackets with variable defined as the index. ", "\nI have tried the following as decribed in the code\nset p \"not r\\[0\\]\"\nset bit 0\nset value 1\nset x [regsub -all {r\\[$bit\\]} $p $value] \n\nExpected result is x = \"not 1\"\nOutput = 0\n\nA:\n\nThe reason why there is no substitution is because you the expression is trying to match the literal string r[, then $ which means 'end of line', then the literal string bit]. ", "To allow substitution of variables, you will have to use double quotes, but that means you will have to be more careful with the escaping since with that, you now have to double escape (one to escape the backslash, another to escape the square parens):\nset p \"not r\\[0\\]\"\nset bit 0\nset value 1\nset x [regsub -all \"r\\\\\\[$bit\\\\\\]\" $p $value] \n\nThe above gives you not 1.", "\nSome not so common solution I guess would be to try using format to perform the substitution:\nset x [regsub -all [format {r\\[%s\\]} $bit] $p $value] \n\nOr the string literal metacharacter (***=) for Tcl with double quotes:\nset x [regsub -all \"***=r\\[$bit\\]\" $p $value] \n\n" ]

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[ "Maybe CNN lacks the guts to directly claim that a Republican sent those devices. ", "And so it has resorted to the cowardly contrivance of putting the accusation in the mouths of others.", "\n\nFirst, in its 6 AM New Day opening this morning, Alisyn Camerota highlighted a Washington Post headline: \"Amid Incendiary Rhetoric, Targets of Trump's Words Become Targets of Bombs.\"", "\n\nJust in case the accusation wasn't explicit enough, CNN—under a \"Breaking News\" banner—then ran a clip of New York Gov. Andrew Cuomo [brother of CNN's Chris Cuomo] saying:\n\n\"This is political terrorism. ", "This is American terrorism. ", "This is red versus blue terrorism.\"", "\n\n\"Red versus blue terrorism.\" ", "Got that? ", "Cuomo unequivocally claims that this is a case of Republican terrorism targeting Democrats. ", "And CNN runs that as Breaking News.", "\n\nThe WashPost story, by Philip Rucker, reads like a sloppy editorial. ", "It begins:\n\nIn the home stretch of the fall campaign, President Trump has called Democrats “evil” and argued they are “too dangerous to govern.” ", "He has denounced Barack Obama’s presidency and demonized former secretary of state Hillary Clinton, inspiring chants of “Lock her up!” ", "at his rallies. ", "The president has also used his bully pulpit to taunt Rep. Maxine Waters (D-Calif.) as a “low I.Q. individual,” impugn former CIA director John Brennan and fan conspiracy theories about liberal philanthropist George Soros. ", "And he has called the news media “the enemy of the people,” singling out CNN’s reporting as “fake news.” ", "This week, these targets of Trump’s rhetoric became the intended targets of actual violence in the form of pipe bombs, many of which turned up Wednesday.", "\n\nNote: Cuomo also seems to be suffering from a bad case of pipe-bomb envy.", "\n\n\n\n" ]

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[ "Royal Oak attorney had sex with juvenile call-girl, Feds say\n\nA Royal Oak attorney is jailed without bond in a case where federal investigators charge he began having sex with a prostitute when she was 14 years old in exchange for legal representation, gifts and cash.", "\n\nDocuments filed in federal court in Detroit show Donald Guest, 43, met the girl in 2010 shortly after she began advertising as an escort on Backpage.com and had a two-year relationship with her that continued even after he was arrested.", "\n\nPhone records indicate Guest was obsessive about calling the girl, referred to as “T.P” in court documents, dialing her number 1,364 times in the first four months he knew her and more than 1,700 times from April to Oct. 27, 2012, the day she was picked up by federal authorities and interviewed.", "\n\nAdvertisement\n\n“The sheer volume of communications that Guest sent to T.P. (shows) an intent to stalk or control her in some fashion,” U.S. Magistrate Judge David Grand wrote Aug. 11 when he ordered Guest jailed without bond while awaiting trial.", "\n\nGrand said the evidence against Guest is overwhelming and shows he engaged in “manipulative, coercive and threatening behavior” to maintain his relationship with the girl.", "\n\n“More recent evidence shows that Guest is unable to control his criminal behavior,” the magistrate judge wrote, adding that Guest was a flight risk who had recently taken an “odd … trip to Uganda.”", "\n\nGuest’s wife appeared in the courtroom during her husband’s hearing before Grant, but her presence – along with Guest’s strong ties to the area and lack of previous criminal history – was not enough to convince Grant that the attorney presented no flight risk.", "\n\n“Not only did Guest carry on a lengthy sexual relationship with a young minor, but he represented her in exchange for sex, threatened to cease representing her if she did not provide him with sex, and (approved of) her providing an alias to authorities,” Grant wrote. “", "Grant also could not control his criminal sexual behavior even after he had been arrested.”", "\n\nGuest’s world began to fall apart the day agents with the federal Southeastern Michigan Crimes Against Children Task Force recovered T.P. on Oct. 27, 2012. ", "The girl, by then 17, told an FBI agent that Guest was her attorney and one of her clients. ", "Guest was arrested the same day on criminal sex charges.", "\n\nThe girl told authorities that Guest was one of her first prostitution customers. ", "He paid her $100 for sex dates, took her shopping and paid for her to have her hair styled, according to an FBI agent’s affidavit filed in federal court earlier this month.", "\n\n“T.P. recalled that guest took her shopping at Northland Mall for her 15th birthday and gave her cash for 16th birthday,” the agent wrote.", "\n\nOne text message exchange between Guest and the girl showed how their legal and sexual affairs sometimes played out, court documents show.", "\n\nGuest: “Were you gonna see me?”", "\n\nT.P.: “Y not”\n\nGuest: “Idk (I didn’t know) I was just asking cause u said after we finished up in court you would.”", "\n\nT.P.: “U got money right”\n\nGuest: “I’m talking about for helping you out. ", "Remember?”", "\n\nGuest: “Just get the appeal form from your new lawyer. ", "I knew you would go back on your word.”", "\n\nAfter authorities arrested Guest, he reportedly admitted to investigators that he engaged in sex with T.P. and her cousin T.M. who also worked as a juvenile prostitute, court records show.", "\n\nHe represented T.P. on a prostitution charge in Warren in 2012 and was able to get the charge reduced to solicitation. ", "Guest also represented the cousin when she faced charges related to a school fight, according to court documents.", "\n\nIn T.P’s Warren case, where she gave police officers an adult alias, Guest wrote in a text: “And its not even under your real name so it don’t matter.”", "\n\nThe federal charge against Guest is for transporting a juvenile across state lines with the intent to engage in criminal sexual activity. ", "The charge stems from a trip authorities say Guest took with T.P. in July 2012 to Georgia. ", "The girl was arrested there for providing police with false identification. ", "She left some of her belongings in an impounded vehicle.", "\n\nShe was released and returned to Detroit. ", "Two weeks later, Guest drove her to Georgia, records show, in exchange for sex so she could retrieve her belongings.", "\n\nThe State of Michigan Attorney Discipline Board disbarred Guest in January 2004 for mishandling money from court judgments, putting them into his personal checking account and forging checks on his employer’s account. ", "His license was reinstated in May 2010." ]

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[ "~ Musings of a gay ex-Mormon father\n\nMy First Boob Was at a Drive-in Movie\n\nI’ve decided to republish my all time top 5 most visited posts. ", "Here’s #5…\n\n_______________________\n\nDrive-in movies seem so 1950 -1970. ", "I’m sure I’m not the only boy who saw, touched or felt his first breast at one. ", "But mine didn’t happen in the typical teenager-making-out-in-the-car sort of way that you’d imagine.", "\n\nWhen I was a child and one of 8 kids in the family, drive-in movies were the only movies that my parents could afford to take us all to. ", "You paid per car back then and got a double feature. ", "We’d all dress up in our pajamas and bring our pillows. ", "Mom would pop popcorn and put together bags of treats. ", "My parents would load their stash of Tab or Fresca sodas. ", "We’d all pile in the station wagon and head to the drive-in at dusk. ", "I loved everything about these adventures.", "\n\nEven when I was teaching ESL in the 90’s, my foreign students thought drive-in movies were the epitome of American culture and made sure it was on their bucket list of things to do in the USA.", "\n\nMost Americans now are unaware that the tradition lives on. ", "There are still open and active drive-in movie theaters. ", "I never go on my own or with other adults. ", "But as a dad, they are still one of my favorite things to do with the kids… just like when I was the 10 year old boy in Six Million Dollar Man pajamas.", "\n\nIn fact they’ve gotten better. ", "Instead of the old mono speaker patrons used to hang from their windows, theater-goers now tune in to a local FM station and can hear the movie on the car stereo. ", "I have an SUV that I face backwards in the rear and make a nice bed in the back with all the seats down. ", "We also bring camping chairs and some of us sit outside with the doors open.", "\n\nLike my Mom, I bring along bags of treats for the kids. ", "The kids wear their pajamas and there are lots of pillows and blankets involved. ", "On the way, I usually grab burritos or a pizza for dinner and get there early.", "\n\nThe younger kids are usually asleep by the time the second movie starts so it seems obvious that the first movie should be a fun family movie and the second one something that the parents and older kids would enjoy. ", "It rarely works like that nowadays but I seem to remember it that way… watching a Disney movie and then being told to go to sleep while my parents watched something racier.", "\n\nI remember seeing my first onscreen boob at a drive-in by not falling asleep for the second movie. ", "I have no idea what the movie was, but I recall the bare breasts.", "\n\nSeeing tits didn’t do anything for me even back then and it would have been a non-event for me except for my mother’s reaction. ", "She was indignant and got upset at my Dad for some reason…perhaps he had been called in to advise the director of this movie? ", "I don’t know. ", "Who knows what craziness goes through a Mormon mother’s mind when flesh is revealed. ", "I’m guessing that it was my dad who selected the movie and so he was to blame for its contents. ", "My mother’s freaking out about it only further solidified my first impression that this was something I should NOT be seeing… ever.", "\n\nTo tell the truth, the actual movie is secondary to the whole experience anyway. ", "Yesterday the kids asked to go see a drive-in movie this weekend and I said OK without even knowing what’s playing. ", "I can’t even tell you one drive-in movie I saw as a kid although I’d guess a Herbie the Lovebug or Apple Dumpling Gang sequel was thrown in there somewhere.", "\n\nBut the pair of boobs, those I remember.", "\n\nNow, I’m not one to take my kids R rated movies but ratings now and ratings back then are two different things. ", "I’ve rented old PG movies for my kids that include enough violence and sex that would be R today. ", "But I wouldn’t freak out if my kids saw flesh onscreen. ", "What would probably happen is this…. ", "the kids would scream, “GROSS!!” ", "or “Yuck!” ", "or “Disgusting!”", "\n\nAfter 10 years of living with their mother who, like my own mother, wasn’t on good terms with her own flesh let alone the flesh of others, this sort of reaction bugs me. ", "So, I’d probably interject with something like, “That’s not disgusting. ", "It’s someone’s body. ", "It’s inapropriate maybe, but it’s not disgusting.” ", "In fact I’m fairly certain I’ve said that enough in other situations that my kids could probably repeat the speech on their own.", "\n\nI’m just trying to help them appreciate and not fear their own bodies. ", "I’d rather that they recoil at unkindness and hypocrisy than at human flesh.", "\n\nReally? “", "Tit” is offensive? ", "Why is it any worse than “boob” or “breast”? ", "And what does my being gay have to do with it?", "\n\nThe intent was to be irreverent and bring people in to read it. ", "And it seems like it worked in your case. ", "You clicked on it.", "\n\nI actually think your reaction is exactly what I was talking about in the post…freaking out over trite issues regarding our bodies indicates a fear and loathing of the human body. ", "Things that you find disgusting need to be glossed over with pretty names. ", "If you were OK with it, it wouldn’t matter what it was called…\n\nStill, I realize that hypersensitive folks will visit here too, so I’ll change it for you all…" ]

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[ "(ANSA) - NEW YORK, 1 GEN - Compiva quel giorno 15 anni e fu la prima a sbarcare: Capodanno 1892. \"", "Avanti le donne e i bambini\": Annie Moore, partita dall'Irlanda 12 giorni prima, fu la prima dei passeggeri della sua nave a transitare per la nuova stazione di ispezioni federale nel porto di New York. ", "Divisa tra chi crede nei muri e chi cerca di costruire ponti, l'America oggi ha celebrato l'anniversario. ", "All'arrivo la \"migrante zero\" ricevette una moneta d'oro: \"Dice che non se ne separerà mai.", "\n\nChe la terrà sempre per ricordo\", scrisse 125 anni fa il New York Times registrando l'avvenimento. ", "Contro le due ore e passa di un passeggero che arriva oggi a JFK, Annie e i due fratelli minori Anthony e Phillip impiegarono mezz'ora a superare i varchi di quella che poi venne battezzata la \"Porta d'Oro\" e riabbracciare i genitori che abitavano a Manhattan da qualche anno, ma a loro fu riservato un trattamento da Vip. ", "Dopo Annie e fino al 1954, 12 milioni di immigranti sfilarono davanti ai funzionari di Ellis Island.", "\n\n" ]

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[ "I really like Charjabug.", "\n\nNot having proper blue/green for Vikavolt sucks, but it turned out OK." ]

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[ "Q:\n\nAsp.net Timer resets on page refresh\n\nI'm using a simple asp.net timer with this code as a backend \nprotected void Timer1_Tick(object sender, EventArgs e)\n{\n int seconds = int.", "Parse(Label1.Text);\n if (seconds > 0)\n Label1.Text = (seconds - 1).ToString();\n else\n Timer1.Enabled = false;\n}\n\nThe timer is grouped along with a Label and a Button inside a UpdatePanel in my front-end. ", "\n<asp:ScriptManager ID=\"ScriptManager1\" runat=\"server\"></asp:ScriptManager>\n<asp:UpdatePanel ID=\"UpdatePanel1\" runat=\"server\">\n <ContentTemplate>\n <asp:Image ID=\"RedBox\" runat=\"server\" ImageUrl=\"~/images/redbox.png\" Visible=\"false\" CssClass=\"redbox1\" ClientIDMode=\"Static\" />\n <asp:Button ID=\"schlbtn1\" runat=\"server\" Text=\"Go To Lectures\" CssClass=\"btn btn-lg btn-success btn-block\" OnClick=\"schlbtn1_Click\" Visible=\"true\" ForeColor=\"Black\" ViewStateMode=\"Enabled\" ClientIDMode=\"Static\" />\n <asp:Label ID=\"Label1\" runat=\"server\" Visible=\"false\" CssClass=\"timerlbl\" Font-Size=\"XX-Large\">60</asp:Label>\n <asp:Timer ID=\"Timer1\" runat=\"server\" Interval=\"1000\" OnTick=\"Timer1_Tick\" Enabled=\"false\">\n </asp:Timer>\n </ContentTemplate>\n</asp:UpdatePanel>\n\nI'm looking for a way to make the timer NOT to reset on every page refresh. ", "I need the timer to still count down until it reach 0 even if the client is not on the same page. ", "I tried Sleep methods but think this is not the solution. ", "Please make any kind of suggestions.", "\n\nA:\n\nStore the timeout in the session:\nAdd to page load:\nif (Session[\"timer\"] == null)\n{\n InitTimer(120); // or whatever initial value is\n}\n\nAdd method:\npublic void InitTimer(int secs)\n{\n Session.", "Add(\"timer\", secs);\n Timer1.Enabled = true;\n}\n\nChange tick method:\nprotected void Timer1_Tick(object sender, EventArgs e)\n{\n int seconds = (int)Session(\"timer\");\n if (seconds > 0)\n {\n seconds--;\n Session[\"timer\"] = seconds;\n Label1.Text = seconds.", "ToString();\n }\n else\n Timer1.Enabled = false;\n}\n\n" ]

{ "pile_set_name": "StackExchange" }

[ 0.001175995683297515, 0.0010362733155488968, 0.0016212265472859144, 0.0008548647165298462, 0.000772052095271647, 0.0005765894893556833, 0.0009988881647586823, 0.0008661719621159136, 0.0010891814017668366 ]

[ "/**\r\n * @author: salimt\r\n */\r\n\r\nimport edu.duke.*;", "\r\nimport java.util.*;", "\r\n\r\npublic class WordFrequenciesMap {\r\n\r\n public static void main(String[] args) {\r\n tester();\r\n }\r\n\r\n public static void countWords(String filename){\r\n FileResource fr = new FileResource(filename);\r\n ArrayList<String> words = new ArrayList<String>();\r\n ArrayList<Integer> counters = new ArrayList<Integer>();\r\n for(String w : fr.words()){\r\n w = w.toLowerCase();\r\n int index = words.indexOf(w);\r\n if (index == -1){\r\n words.add(w);\r\n counters.add(1);\r\n }\r\n else {\r\n int value = counters.get(index);\r\n counters.set(index, value + 1);\r\n }\r\n }\r\n int total = 0;\r\n for(int k=0; k < words.size(); k++){\r\n if (counters.get(k) > 500){\r\n System.out.println(counters.get(k)+\"\\t\"+words.get(k));\r\n }\r\n total += counters.get(k);\r\n }\r\n System.out.println(\"total count: \"+total+\" different = \"+words.size());\r\n }\r\n\r\n public static void countWordsMap(String filename){\r\n FileResource fr = new FileResource(filename);\r\n HashMap<String,Integer> map = new HashMap<String,Integer>();\r\n\r\n for(String w : fr.words()){\r\n w = w.toLowerCase();\r\n if (!", "map.containsKey(w)){\r\n map.put(w,1);\r\n }\r\n else {\r\n map.put(w,map.get(w)+1);\r\n }\r\n }\r\n int total = 0;\r\n for(String w : map.keySet()){\r\n int value = map.get(w);\r\n if (value > 500){\r\n System.out.println(value+\"\\t\"+w);\r\n }\r\n total += value;\r\n }\r\n System.out.println(\"total count: \"+total+\" different = \"+map.keySet().size());\r\n }\r\n\r\n public static void tester(){\r\n String filename = \"data/confucius.txt\";\r\n double start = System.currentTimeMillis();\r\n countWords(filename);\r\n double end = System.currentTimeMillis();\r\n double time = (end-start)/1000;\r\n System.out.println(\"time = \"+time);\r\n start = System.currentTimeMillis();\r\n countWordsMap(filename);\r\n end = System.currentTimeMillis();\r\n time = (end-start)/1000;\r\n System.out.println(\"time = \"+time);\r\n }\r\n\r\n}\r\n" ]

{ "pile_set_name": "Github" }

[ 0.0009057869901880622, 0.0008126182947307825, 0.002392822178080678, 0.0025109117850661278 ]

[ "We stand on the far promontory of centuries!What is the use of looking behind ussince our task is to smashthe mysterious portals of the impossible?", "\n\n—“Futuristic Manifesto”, Filippo Tommaso Marinetti~~~\n\nEverything’s progression. ", "You and me,\nwe’re both respective tips of blades\nmade sharp—or dull— by “Father looked\nat life like this, and mother that, so I…”\nAnd if we die sans heirs, we all still\ninfluence. ", "Creation grinds our vagaries\nto dust beneath my feet and frees\nme from the appetite to disagree.", "\nKeep up or don’t. ", "Like Etruscan sentries,we stand on the far promontory of centuries,\n\ncontributing with earthy bits and pieces\nto terroir that grows a wine particular\nto you, I find abrasive or a sickly sweet,\nand yet, I’ll creep at night to taste again, in case\nI missed some subtlety, and by a single peep\nmy concave/convex lens adjusts. ", "It grinds us\ninto sharper focus or like plates tectonic\ngrates and makes distinct new continents:\nPangaea, panacea, panegyrics, all blinds us.", "What is the use of looking behind us\n\nif dread and praise have lock stepped\nso that nothing good I say to you\nis heard, and every unintended slight\ncuts to the bone? ", "We’ve split apart\nand there’s a fact that oceans of affinity\nwill never trouble to correct. ", "You dot, I dash,\nwe are a code no more in vogue, a set\nof peeves like kitchen knives whose history\nprovokes no interest, even less of cash.", "Since our task is to smash,\n\nas far as I can tell, the misbelievers\nof their woebegotten truths so there’ll be\nless of them, it stands to reason that by leaving you\nto tilt your mills and me to grind my axes,\nsome third construct of our selves will\ncircumlocute to an axis made more plausible,\ndare I say fun, with extra-sensate lubricants.", "\nMeanwhile, the new year, like a chariot, rolls in,\nits wheels, friction-free, making audiblethe mysterious portals of the impossible.", "\n\n~~~\n\nFilippo Marinetti (1876-1944) was a poet and founder of the Italian Futurist movement. ", "His work is brash and energetic and crackles with outrage. ", "If Marinetti were alive today, social media would be all over him, and we’d be making or breaking friendships based on our alliance. ", "Love it or loathe it, Post-modernism, too, will be history one day.", "\n\nThe image comes from a deck of inspirational cards called Art Oracles. ", "This glosa proves to me they work.", "\n\nGive me your hand and we will dance;\ngive me your hand and you will love me.", "\nLike a single flower we will be,\nlike a single flower, nothing more.", "\n\n—“Give Me Your Hand” (“Dáme la Mano”) by Gabriela Mistral\n\nI dreamed of a friend in an orange checkered suit,\ngarish, clashing patterns, layered shades of yolk.", "\nHe milled, a hydrant, awkward midst the artsy party crowd.", "\nMortified, I hissed: why are you here?", "\nHe brightened. ", "I’ve looked everywhere!", "\nI thought I’d lost my chance.", "\nThe places I frequent are thin in godless times;\nto be Olympian, hope and patience teeter.", "\nBut enough of that. ", "Do you like my pants?Give me your hand and we will dance.", "\n\nHe drew the blinds and took me in his arms.", "\nI do not know the steps, I whined, and shuffled stiff.", "\nThey’re easy, he replied, though I often wonder if\nthe laurels people hang on strife\nand being an enduring wife or husband\nhave not muddied things a bit. ", "You see,\nI do not need a maid and trust\nyou’ve had enough of joyless handymen\nwho’d nail your freedom to a tree.", "Give me your hand and you will love me.", "\n\nIn time, my limbs began to melt\nand I misplaced embarrassment. ", "He led,\nnot like a general or a cold front pushing through\nbut like the tall straight mast of a merchant\nsailing ship, with goods fair traded\nin his hold. ", "I think that we shall be,\nhe whispered in my ear, a golden pair\nwell matched, unfolding like the petals\nof a rose, unprecedent, named Liberty.", "Like a single flower we will be.", "\n\nWe woke entangled in a king-size bed\nin Tuscany beneath an arbour\nwoven with bay laurel and anemone.", "\nIt must be spring, I reasoned, peering\n‘neath the sheets at what he’d brought.", "\nA lot! ", "We laughed from bed to floor\nand rolled across to where our view\nof self-created destiny was clear.", "\nWe’d risen, both, to all that we adorelike a single flower, and nothing more.", "\n\nNot long ago, I found a strange map\nin the ruins of a Maracaibo mansion,\nthe corners held down with rough-cut rubies\nround and plump as duck eggs. ", "Palimpsests\nthrobbed like blue-black veins across the chart—\nillegible, unscarred by zealots and thieves.", "\nI was told by the raggedy viejo who sleeps\nunderneath that the map and her routes\ncan be viewed by whoever believesthe jaguar brushed leaves\n\nwith her tail and the weasely dictator fell.", "\nClaims such as these, they never sit well\nwith the rushed and the rational. ", "Being neither\nthat day, I asked the old man to explain.", "\nOnce a year, he said, when defenses\ndeflate, humankind’s natural omniscience\nis recalled and recorded upon this map\nby shades of the recently departed who’ve\ndropped all pretence of sorrow and vehemence.", "With his phosphorescent absence\n\nof political skews and racial miscues,\nhe hovered over the map, and with a finger\ngnarled as ebony burl, he cruised along\nroutes I’d been known to frequent and\nrubbed them all out, pronouncing every one\nirrelevant. ", "Time to accept there’s no fact in\nthe past with the power to deplete or subvert\nyour future. ", "Take a page from the wild. ", "When\nthe cayman’s not hungry, he’s loath to attack, andthe puma speeds through bracken.", "\n\nLikewise, in the seam between moments—and\nyears—that appear to engender and justify\nfear, you will find a clear trail laid out by the good\nthat is you and your boundaryless kin. ", "You are\ntimely, well compassed. ", "Walk on, begin.", "\nAnd now it is time for this Job to expire.", "\nHe dropped the fat rubies into a sack.", "\nHe rolled up the mansion and with it the map,\nspinning all he had shown me into a gyrelike devouring fire…\n\nI’ve been chewing darkness for so long\nthat I don’t know how to relearn joy;\nI’ve been walking on lava for so many years\nthat my feet have lost all memory of fleece.", "\n\n—Gabriela Mistral, “Nocturne of Consummation”\n\n~~~\n\nI made away today to the blue green\nwaters, slipping through crevasses of\nmusty inattention, past the trawling clusters\nof opinionates despairing of their worlds\nto reach this remnant of an iron age. ", "I used to wait\non people I adored who saw in everything a wrong.", "\nI danced my favourite red shoes off to prove the right\nand bled in endless causes. ", "Nearly lost my head\nyearning, fruitless, where I never could belong.", "I’ve been chewing darkness for so long\n\nthe fife bands of the mind with their penny\nwhistle tweedling, their tinpot repercussion\nof past victimhoods march at my heels, wanting\nme to swallow pain as good for me, agree to being\nground to ash until my joints and sinews ache.", "\nNothing tastes the way it did when I enjoyed\nlife, which qualifies my joining the insipid who bash\ntheir heads against the walls of Plato’s cave. ", "Their aim\nis group concussion, so torpid and pointless a ploythat I don’t know how to relearn joy.", "\n\nAnd yet, now that I’m here\nwith only forward as my guide\nand no convincing evidence that death\nlike the dusty fly-bit reign of Ozymandias\nis worse or better than anything else,\nI can turn to the cantankerous my deaf ears,\nleave one-trick ponies to their sad politics,\nappreciate the strides humanity has made,\nhas yet to make beyond the vale of fears.", "I’ve been walking on lava for so many years\n\nthey know my name in Pompeii;\nPopocatepetl is my winter home.", "\nBut for all the pumice I’ve endured,\nthe present me sees fresh at every turn\nand boredom as the only borderlands.", "\nProsperous, sans alms or palms to grease,\nI welcome the agora surrounding me\nwhose wares and wherewithals are\nso abundant, true, intent on peacethat my feet have lost all memory of fleece.", "\n\nFree thinker! ", "Do you think you are the only thinker\non this earth in which life blazes inside all things?", "\nYour liberty does what it wishes with the power it controls,\nbut when you gather to plan, the universe is not there.", "\n\nI have a shelf in a quiet corner\nof my house where books appear,\nwhere spines with startling titles\nwink like coin upon a beach\nyou think, at first, is crumpled foil.", "\nI move in close. ", "You little stinker,\nwhere’d you come from? ", "Didn’t I just\ndust here yesterday? ", "Three Bly collections,\nnow there’s four! ", "Best not to blink or—free thinker! ", "Do you think you are the only thinker?", "\n\nLaughter of the cynical sounds hardy\nlike a whack across the shoulder blades.", "\nWell met, friend! ", "One scarcely hears\nthe swallowed—gullible—or feels\nthe poisoned tip of reason penetrate.", "\nThe spy pretends to care. ", "He brings\nhis little eye of mean intelligence,\nthen shrinks. ", "He’s leather in the rain.", "\nI know him well. ", "I know he stingson this earth in which life blazes inside all things.", "\n\nMeanwhile, this new-found book,\nthe fourth or maybe fifth this year\nfalls open as do all things freshly\nmanifest, and from its novel pages\npour like immigrants through Ellis\nand old Halifax onto these shoals\nnew habitants, thoughts never known.", "\nTides reverse. ", "My salty backward-facing pillars\nburble, angry, smash their begging bowls.", "Your liberty does what it wishes with the power it controls,\n\nThe pale brittle shell of politics is broken.", "\nNewborn patriots stand blinking in a sun\nthat’s never shone like this till now.", "\nThe rush of sea, the boats well laden\nwith supplies will dry all eyes\nonce sorrowful. ", "You who swear\nthe age of miracles is dead, you’ll find your proofs.", "\nWho negates life for afterlife, division\nas your goal, we too will meet somewhere.", "But when you gather to plan, the Universe is not there.", "\n\n~~~\n\nIt’s been a while since I’ve written a glosa, but even longer since I’ve felt proud of my nation and government down to the cellular level. ", "There’s been a sea change in Canada since the election of Prime Minister Justin Trudeau. ", "I see young people politically engaged for the first time. ", "American friends post videos of our PM greeting new arrivals and say they feel the shift too, even through all their media clamour.", "\n\nIf glosas and form poetry appeal to you, please visit my website where you can learn more–and perhaps buy a book or two.", "\n\nA tree of blood moistens the morningwhere the new mother groans.", "\nHer voice leaves crystals in the wound\nand a diagram of bone in the window.", "\n\nMeanwhile the coming light holds steady\nand overtakes blank limits of fable that forget\nthe tumult of veins in its flight\ntoward the turbid cool of the apple.", "\n\n–“Adam”, from Federico Garcia Lorca’s Primeras Canciones, 1922\n\n~~~\n\nThe drop of ink that falls\nabsorbed between the fibers\nof a parchment bed no pen\nof yours or mine can resurrect\nis spreading. ", "Footprints of tar\npace a figure eight, delineating\nnakedness that none of us can see\nexcept in vaguely worded fantasies.", "\nDebris across the mountain feels like warning;a tree of blood moistens the morning\n\nwhile I am still senseless\nand too sensitive, I can refrain\nfrom driving home some vagrant\npoint of fact no one has invited,\nleast of all that dead bore couple,\nhabit and experience. ", "Phones\nattached to hips are gathering\nlone gods of randomness in droves.", "\nBeggars with no credit offer loanswhere the new mother groans\n\nlabouring to no avail, you’d think,\nwould set off some alarm—what child\nis this? ", "But no one’s claiming fatherhood,\nand that fat bastard capital forgets\nto keep his mouth closed when he chews.", "\nWhat trickles from his chin is spooned\ninto tubes, shot straight into the veins\nof pretenders to Cassandra, whose Trojan\nnever breaks and is still well tuned.", "Her voice leaves crystals in the wound\n\nthat rub against synthetic outrage\nwaiting for its moment—that will never\ncome—of sweet approval. ", "What tendency\nis this to sprinkle vinegar upon a neighbour’s\nolive grove that looks to be abandoned?", "\nNo succulent upon a fence can grow\nwhen roots are parched of laissez-faire.", "\nWe subatomic dancers hate rehearsal,\nswiftly leave behind our sold-out showand a diagram of bone in the window.", "\n\nSo what did the rich man say\nto the ferryman? ", "I’ll be damned!", "\nOnly he wasn’t and still the river\nfoams in hopes that someone might\napproach her self-creation\nin a feathered cape with dignity.", "\nSir Walter and the puddle knew\nGood Bess was on her way. ", "All others\nin the fractious crowd stayed petty.", "Meanwhile the coming light holds steady\n\nand the errant ink grows jittery\nfor having glimpsed the perfect\nquill in V-formation flying over\nParry Sound. ", "What if I dry\nand flake apart before we two\ncan prove the world is wet?", "\nThe goose without a fleeting honk\nflies on. ", "She does not give her tail lightly.", "\nEggs of gold each day she brings to marketand overtakes blank limits of fable that forget.", "\n\nTwo things depreciate at the moment\nof purchase: the second is worry.", "\nGrinding mandibles on behalf of another\nforetells a long decline toward mush\nand not much else. ", "No imagination\nwill fling you out of Eden. ", "Paths of right\nand wrong confuse the tenant farmer,\nnot the lord who views all he surveys\nwith potential, green and bright,the tumult of veins in its flight.", "\n\nOh, sweet desire, now that you know\nmy name, let’s draw the canopy against\ndrunk beetles banging on their broken\nschemes. ", "Not all shells are suitable\nfor dyeing, though every word, I’m told,\nwill find its violin and grapple\nfor the pitch it hears in dreams\nof paradise, giving way to the refugee,\nnourished in his flight by sunlight’s dappletoward the turbid cool of the apple.", "\n\n~~~\n\nThis glosa borrows the first two stanzas of Garcia Lorca’s poem, “Adan”. ", "The original Spanish can be found here. ", "I’ve published a book of glosas, Dead to Rights, with an accompanying novella, Dead Edit Redo, which you can find here.", "\n\nHe wrapped her warm in his seaman’s coat against the stinging blast; he cut a rope from a broken spar and bound her to the mast.", "\n\n—“The Wreck of the Hesperus”, Henry Wadsworth Longfellow\n\nIn the moribund night of a waning moon\non the crags of an island known as Doon\no’ Fara, moves the shadow of a weaver\nfrom thatchéd hut to cliffs of spray and salt.", "\nBy day she spins and knits complicated\ngarments for the discerning and remote.", "\nBy all accounts, her wealth cannot be touched\nor measured, though she started life as\nproperty of one they called the Stoat.", "He wrapped her warm in his seaman’s coat\n\nand circled her, resentful, day and night.", "\nWhate’er she thought or dreamed, he knew.", "\nHe brought her sprigs of violet and skeins\nof tangled wool to while away her hours.", "\nHe filled her head with tales of dread and\ndisappointment that sealed her like a cast.", "\nYou are my legacy, he’d croon. ", "When I am gone,\nyou’ll carry on my song of life’s depravity,\nwrapped firmly in the wisdom of my pastagainst the stinging blast.", "\n\nThe weaselly man he traveled far, indulging\nendless appetites. ", "To ease his welcome home,\nhe filled the holds of ships with ivory bits and\nwooden masks in such vast quantities that\nFara could not move inside her thatchéd prison.", "\nSome folk say she clubbed him with a bar\nof solid gold; others say he met his end\nin polar realms—who knows? ", "One day,\nshe hired a young man home from war.", "He cut a rope from a broken spar\n\nand built a sledge, and together they expunged\nall traces of the dark controlling Stoat. ", "With every\ndiscard off the cliffs, her mind became more spacious.", "\nThe young man went his way, and she, devoted\nto the doon, mastered patterns of abundance from\nthe roiling wind and sea. ", "Eons since have passed,\nand only in the darkening moon are glimpses of the\nweaver seen. ", "But on certain icy twilights, you may\ncatch the whiff of him who, loathing freedom, castand bound her to the mast.", "\n\ncontact between you &\nGod means no church,\nno society, no reform,\n& almost no relationships,\n& almost no hope in\nrelationships—but\nkindness of hope inherent\nin that what is good,\nshall live & what is\nbad, dies—Your\nflesh will be a husk,\nbut yr. ", "soul a star—\n\n—from Jack Kerouac, Book of Sketches, Oct. 31, 1952\n\n~~~\n\nI\n\nI see you’ve lined up\nyr bottles for tonight’s\nobliteration, hoping for a\nhit, some kind of catapult,\nanother dis-appointment with\nold friends & lovers banned\nfrom sobriety. ", "Let me tell you\nnow, friend, there are no\nlost amigos, only grandcontact between you &\n\nincomplete, you say,\nwithout that special someone\nevery day a seek\n& almost find, that time\nin Jalisco if you hadn’t been\nblind drunk, the search\nwould be long over, yeah?", "\ninsatiable she was,\nonly her crucifix made you lurchGod means no church\n\nbut enough about me—\nhow’s life treating you?", "\nHave you learned yet\nto take a compliment,\nor does that old leather\nstrop torn\nfrom its nail in the woodshed\nstill reek of bay rum?", "\ndo you still bleed his scorn?no society, no reform\n\nI’ve been there, I know.", "\nAll that shit between pretty\ncovers with my name\non it, I scrounged hard\nfrom railyards—rusty\nI-beams and wet pine chips\nfor a bed, alone\nnow I’m practically a saint—\nall those women, a few drips,& almost no relationships\n\nII\n\nso here’s the thing—\nthere are no dead poets\nthere is no dead anything\nsure, mountains are melting\n& a certain green salamander\nwon’t be unfurling her thin\npink tongue for termites anymore—\nbut it’s not yr doing; she’s done\nhere for now, a slow grin& almost no hope in\n\nwishing your fellow man\nwere not so immune\nto yr discontent.", "\nLife seems easier\nwhen you can stir up\nguilt in yr little grass hut\nlike a pot of beans\non low simmer—\nhell, you can’t shutrelationships—but\n\nyou can confuse yourself\nover & over like those old wooden\npaddle toys with rubber ball\nattached—k’bonk, k’bonk, k’bonk,\nwhat’d I do? ", "what’d I do? ", "what’d I do?", "\nshort answer: nothing. ", "A fine gent\nyou are, always will be.", "\nInner space, same as outer,\nnothing lost, mis- or unspent,kindness of hope inherent\n\nbut you’ve heard all this—\nsmoked it & wrote it\n& sold it to a few\nworse off than you. ", "Maybe\nit’s time to clear off those\nshelves. ", "They’ve been yr hood,\nyr holy armor, for how long now?", "\nNo one wants a soldier with flat\nfeet. ", "Letting go’s the only roodin that what is good\n\nIII\n\nSo. ", "Make friends with emptiness.", "\nYesterday’s om and a planet’s\nworth of mountaintops\nwon’t save yr bored soul.", "\nPractice saying, I am deep\n& meaningful, leave the biz\nof others to others\nnever ask them why\nbelieve love and geniusshall live & what is\n\nis. ", "There’s no other\ntense and no better\nway to let go the tension.", "\nStop gluing name tags\nto intolerance—gluten, lactose—\ngive up keeping score;\neverything you look at\nmultiplies—boom, ka-ta-boom!", "\nignorebad, dies—Your\n\nthe one in charge\nof what comes around\n& who stays away\nbut still,\nwe’re amigos to the end,\nbro, through love & lust—\nthrow out the dishwater\nfrom yr last best date—one day\nyou’ll smell, it’s not a healthy muskyour flesh will be a husk\n\nI have to split soon.", "\nYou got tons of visitors\ncuter than me lined up. ", "I just\ncame to oil yr valves, give\nthe silver in yr irises a gleam.", "\nThe New Year isn’t far;\nit’s continuous New Now. ", "We’ll\nmeet again soon. ", "You’ll see\nthat nothing leaves a scar—but yr soul a star!", "\n\ntrust your heart if the seas catch fire\nlive by love though the stars walk backward\nlosing through you what seemed myself, I find\nselves unimaginably mine, beyond…\n\n“dive for dreams”, e.e. ", "cummings\n\n~~~\n\nThe man with the kind smile is in the news today\nfor shame; I think his mother must be feeling\nterrible. ", "How hard she’s scrubbed what hangs in\npublic view. ", "It’s not her laundry now, nor mine, and\nyet I cannot help but cringe. ", "This brine cooked up\nby marketeers of low repute electrocutes—the wire\nrunning through us all burns hot and white. ", "It rips us\ninto trenches, splitting wrong from everybody’s right.", "\nTo him, the kind one, I offer this: while all seems dire,trust your heart if the seas catch fire.", "\n\nLet’s say, for sake of argument, that an institution\nknows what’s best for you and me between the\nsheets and who we call to warm them. ", "The exit\nsigns are clearly marked; I know the rules,\nyou know who dominates, submits. ", "Our excitation\nknows no bounds…so far, so good, our sword\nplay’s in good fun—what’s this? ", "You run to papa,\nnow? ", "What’s changed? ", "This wasn’t in our game!", "\nThe pain that binds us reappears, inside outward:Live by love, though the stars walk backward.", "\n\nNow we’re raining frogs and razor blades,\nthe “I would nevers” puffing up, their croaks\nif not harmonious, at least, are sanctimonious.", "\nMeanwhile, in stalls high up in marble halls,\nstars of varying repute, nova, dark and dwarf,\nfear toppling like you from their hard-won blind\nheavens. ", "What can I do to extract promises\nfrom the many I have loved most awkwardly?", "\nNot much. ", "I thought I’d left you all behind.", "Losing through you what seemed myself, I find\n\na vast, polluted land that someone promised\nme, I can’t remember when or why. ", "I have no\nmaps or guide. ", "The bondage that delighted\nus, our thumbing nose to pharaohs, seems,\nin retrospect, like sweet repose. ", "The failure\nof success I feared most cruelly has dawned\nan angry sun. ", "What would my father say?", "\nI made him proud and do so, still…perhaps,\nthese plagues I’ll weather and discover fondselves, unimaginably mine, beyond…\n\n~~~\n\nI have taken some liberties with this glosa, a form that many of you may now recognize. ", "The lines I’ve taken from e.e. ", "cummings’ poem, “dive for dreams” are not a consecutive quatrain. ", "I gleaned them from the larger poem in response to a current event, which, in its vividness and universality, addresses us all. ", "Cummings, no stranger to controversy in his lifetime (1894-1962), probably wouldn’t mind.", "\n\nTo be as rich as Croesus, or to live without\na care? ", "These matching oars, once mastered\nby our helmsmen, twin plowshares that rode easy\nin a tiller’s hands, the world has split to either/or.", "\nYour name, dear king—a mockery by tyrants\nand vintners of the sour grape—I call\nin secret, and you never fail to come. ", "I do not ask\npermission of the domed or steepled lot. ", "I let them\nplot and scratch. ", "Your pillow talk I still recall:break loose your trim ship’s hawsers, haul\n\nthe baggage of your past and toss it to the dolphins!", "\nHistory is not, you loved to say, the purview of\nthe winners but the ones afraid of what comes next.", "\nYour counsel to Great Cyrus could not penetrate\nthe circles of disdain and scorn that mottled\nhis fine spirit, though a few did understand.", "\nFreed men and widows, wealthy now beyond imagining,\nyou’ll find them on no Senate floor or king’s list,\nbut they’re teaching younger generations well to handthe anchor from its harbor nest, and stand\n\namong the growing mass who knows there is no\nvictimhood, only the choice of each to limit or allow.", "\nYour tolerance of wealth, my beloved Croesus,\nknows no boundaries. ", "Your opting to ascend\nto legend frees us both. ", "Today, I am no concubine.", "\nNameless as I ever was, you’ll find no grand\nor mawkish monuments to one of thousands who\nadored you, but I know what we achieved, and why\nyou set me on that boat, with Apollo’s helping hand,up into the trade winds off the headland.", "\n\nSo, now I turn to you, dear reader, impatient\nin your search for all that’s new. ", "I see the scimitars\nof doubt you try to hide; I hid them too, until I met\nthe richest king who’d ever lived and walked\nbroad streets absent of poverty, no crime,\nand all deaths natural, in their time. ", "The trails\nwe left are narrow, yes, but clean as an arrow’s arc.", "\nIf you would just give up concern; the king was never\nburned! ", "Your golden talent’s limitless. ", "Forget travails,your woven, patched, and thrice stitched sails.", "\n\nIf you’ve been reading Oceantics for awhile, glosas will be nothing new to you. ", "One of my goals is to restore this glorious Spanish medieval form to appreciative modern audiences. ", "My novella of horror and good medicine, Dead Edit Redo, creeps into the darkest mysteries of the glosa. ", "My compatriot of sorts, Alain C. Dexter, published a whole book of them called Dead to Rights. ", "And while we’re on the topic of self promotion, please take a peek at my newest novel of mystery and magical realism, Daughters of Babylon.", "\n\nNow I should like to give credit to two other poets, without whom this glosa could not have been written. ", "Antipater of Sidon lived in Greece in the 2nd century. ", "His poem, “The Bidding of the Harbor God”, forms the tenth line of every stanza and drives the glosa’s rhyme scheme. ", "The masterful translation is by Sherod Santos, an American poet and author of Greek Lyric Poetry: A New Translation. ", "I’ve included Antipater’s full poem here, Santos’s translation, so you, too, can appreciate the talent of both poets.", "\n\nThe Bidding of the Harbor God\n\nTake your thwarts, oarsmen, it’s time to carve\nnew sea-lanes through the breasting swells.", "\nWild gales no longer avalanche the shoals\nor harrow the rigging of a sail’s nerve,\n\nand already out of mud and clay, swallows\nbuild their jug-nests underneath your eaves.", "\nSo quickly now, before the gulled moon leaves\nits slumberous lightweight in the meadows,\n\nbreak loose your trim ship’s hawsers, haul\nthe anchor from its harbor nest, and stand\nup into the trade winds off the headland\nyour woven, patched, and thrice stitched sails." ]

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[ "Introduction\n============\n\nBreast cancer outcome may be affected by germ-line variants in genes that play a role in DNA damage control and repair such as *TP53*(R72P) and *MDM2*(SNP309) \\[[@B1],[@B2]\\]. ", "The Mdm2 protein is a negative regulator of the tumor suppressor protein p53 \\[[@B3]\\]. ", "The R72P (215G\\>C) polymorphism of the *TP53*gene is located in a proline-rich region of p53 suggested to be required for the growth suppression activity of p53 \\[[@B4]\\] and for its ability to induce apoptosis \\[[@B5]\\]. ", "The two variant protein forms, R72 (arginine) and 72P (proline), have been shown to differ in their biological functions: the R72 variant is a stronger and faster inducer of apoptosis than the 72P variant \\[[@B6],[@B7]\\]. ", "The 72P variant also binds more efficiently to iASPP, an inhibitor of pro-apoptotic function of p53, which may be another reason for the inferiority in apoptosis induction of this variant \\[[@B8]\\]. ", "The 72P variant has been found to be more efficient in inducing cell-cycle arrest \\[[@B7]\\] and DNA repair \\[[@B9]\\] than the R72 variant which may protect tumor from chemotherapy-induced apoptosis.", "\n\nPrevious studies have shown that the R72P polymorphism is not associated with increased breast cancer risk \\[[@B1],[@B10],[@B11]\\]. ", "However, an association of R72P with breast cancer survival has been suggested, though with inconsistent results and possibly only in patients with p53-negative tumors \\[[@B10]-[@B16]\\]. ", "It has also been suggested that patients with the Pro/Pro genotype are less sensitive to anthracycline-based treatment than those with the Arg/Pro or Arg/Arg genotype \\[[@B14],[@B16]\\], in line with the Pro-allele being more efficient in cell-cycle arrest \\[[@B7]\\] and DNA repair \\[[@B9]\\] induction.", "\n\nA common single nucleotide polymorphism in the *MDM2*promoter region, a T to G change at nucleotide 309 in the first intron (-410G\\>T; named SNP309), has been shown to create an improved Sp1 binding site, leading to increased expression of the Mdm2 protein and thus attenuation of the p53 pathway and accelerated tumor formation in individuals carrying a germ-line p53 mutation \\[[@B17]-[@B19]\\]. ", "A number of small studies revealed an inconsistent association between SNP309 and breast cancer risk (see overview in \\[[@B1]\\], and \\[[@B20],[@B21]\\]). ", "However, we have shown in a large pooled analyses of the Breast Cancer Association Consortium series that there is no general association of SNP309 with breast cancer, nor if stratified by estrogen receptor (ER) \\[[@B1]\\].", "\n\nIn two small studies no association between breast cancer survival and *MDM2*SNP309 genotype alone was found \\[[@B13],[@B22]\\]. ", "However, the results of one of those studies suggested a differential effect of *MDM2*SNP309 genotype by tumor p53 status (mutant p53 or aberrant protein expression) on breast cancer survival \\[[@B22]\\]. ", "Though *MDM2*SNP309 has been implicated to affect survival in other tumors (for example, \\[[@B23]\\]), as far as we know there are no other publications on breast cancer outcome and this polymorphism, except for a recent publication in *BRCA1/2*carriers of Ashkenazi origin \\[[@B24]\\]. ", "Our aim was to investigate the combined effects of *MDM2*SNP309 and *TP53*R72P polymorphisms and p53 protein expression on breast cancer survival.", "\n\nMaterials and methods\n=====================\n\nClinico-pathologic data and genotyping\n--------------------------------------\n\nBreast cancer cases from four European studies within the Breast Cancer Association Consortium were included in this analysis (Table [1](#T1){ref-type=\"table\"}) \\[[@B1],[@B25]\\]. ", "Patients that were genotyped for *MDM2*SNP309 and *TP53*R72P from studies with follow-up data were included \\[[@B1]\\]. ", "Patient selection criteria, participation rates and information on the collection of follow-up and clinical data are shown in Table [1](#T1){ref-type=\"table\"}. ", "P53 protein expression data were available for two of the four studies (Table [1](#T1){ref-type=\"table\"}). ", "Immunohistochemical staining of TMA slides was performed with a mouse monoclonal anti-human p53-antibody (DO-7, DAKO) (Table [1](#T1){ref-type=\"table\"}). ", "Missing p53 data could be attributed to missing tumor blocks, loss of cores in the slicing or staining process or cores not containing enough tumor material. ", "P53 protein expression scoring and *MDM2*SNP309 and *TP53*R72P genotyping were performed blinded to the survival status of the patients. ", "Genotyping assays were performed by each group separately \\[[@B1]\\] (see Table [1](#T1){ref-type=\"table\"} for assay description). ", "Primer (and probe) sequences are available from the authors upon request. ", "Methods and results in this paper are reported following the REMARK recommendations \\[[@B26]\\]. ", "All studies were approved by the appropriate (Medical) Ethical Research Committees.", "\n\n###### \n\nCharacteristics of the studies and genotyping assays\n\n -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------\n Contributing studies Design Description of case subjects and ascertainment\\ Participation rates Follow-up P53 IHC\\* **Genotyping platform(s)**\\[[@B38]-[@B40]\\]\n (age range) \n ----------------------------------------------------------------------------------------------- ------------------------------------- -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- ---------------------------------------------------------------------------------------------------------------------------------------------------------- ------------------------------------------------------------------------------------------------------------------------------------ ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------ -------------------------------------------------------------------\n ABCS: Amsterdam Breast Cancer Study, The Netherlands \\[[@B41]\\] Hospital-based consecutive cases All operable breast cancer patients aged \\< 50 years diagnosed 1974-1994 in four Dutch hospitals (Amsterdam and Leiden) (23 to 50 years) All patients with paraffin-embedded tissue blocks available (normal tissue) from the Pathology archives and successful DNA isolation (approximately 85%) Active follow-up through the medical registries and general practitioners By IHC staining of TMAs\\* as previously described \\[[@B25]\\]; p53 positive defined as \\> 10% of cells with positive nuclear staining. ", " Taqman\n\n HABCS: Germany:\\ Hospital-based case-control studies Case patients who received radiotherapy for breast cancer at Hannover Medical School between 1997 and 2003 (27 to 91 years) Approximately 80% of case subjects contacted agreed to give a blood sample Active follow-up at the Department of Radiation Oncology, Hannover Medical School NA Restriction enzyme-based assays\n Hannover Breast Cancer Study and bilateral breast cancer patients \\[[@B42],[@B43]\\] \n\n HEBCS:\\ Hospital-based case-control study Consecutive incident cases from the Department of Oncology, Helsinki University Central Hospital 1997-1998\\ 79% of the case subjects Active follow-up of the medical records until five years and annual linkage to the nation-wide Finnish Cancer Registry By IHC staining of TMAs\\* as previously described \\[[@B10]\\] and data for 23 cases derived from the pathology reports; p53 positive defined as \\> 20% of cells with positive nuclear staining. ", " RFLP (*MDM2*SNP309)\\\n Helsinki Breast Cancer Study \\[[@B10],[@B44]\\] (22 to 96 years) Amplifluor(tm) fluorescent genotyping (Kbiosciences) (*TP53*R72P)\n\n SEARCH: Studies of Epidemiology and Risk Factors in Cancer Heredity, Cambridge, UK \\[[@B45]\\] Population-based case-control study Two groups of case patients (prevalent and incident) identified through East Anglian Cancer Registry: patients diagnosed before age 55 years in 1991 to 1996 and still alive when study started in 1996 and patients diagnosed before age 70 years since 1996 (25 to 65 years) 64% of eligible case subjects provided a blood sample Combination of passive follow-up through national death registrations and active follow up every five years by the cancer registry NA Taqman\n -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------\n\n\\*IHC = immunohistochemistry; TMA = Tissue Micro Array; NA = not applicable (no p53 data available).", "\n\nStatistical analyses\n--------------------\n\nUnivariate analyses of survival were performed by calculating Kaplan-Meier survival curves and comparing subsets of patients using log-rank test. ", "To explore the effects of several variables and their combined effects on survival, multivariate Cox\\'s proportional hazards regression models were used (reported as Hazard Ratio (HR) with 95% confidence interval). ", "Results are reported for one polymorphisms stratified by the other polymorphisms or p53 expression, adjusted for other covariates. ", "Interaction terms were tested by Cox regression models including the main effects (2df each), interaction terms, for example, four interaction terms for both polymorphisms, and other covariates. ", "Covariates included were prognostic factors for breast cancer survival, that is, age, stage, grade and ER and p53 protein expression. ", "In order to run models including all patients, missing value categories were included for each separate variable with missing information. ", "Polymorphisms were included as categorical variables (with the homozygous common allele group as reference), or as a continuous variable in the per-allele analyses. ", "All pooled analyses were adjusted for study, that is, ABCS, HABCS, HEBCS, SEARCH, included as a categorical variable. ", "Breast cancer-specific survival was defined as survival until death from breast cancer, with breast cancer being the underlying cause of death; death due to other causes was censored (these analyses included the ABCS and HEBCS studies, see Table [1](#T1){ref-type=\"table\"}). ", "Overall survival was defined as survival until death of any cause. ", "In all analyses, follow-up time was censored at 10 years. ", "All statistical tests used were two-sided and *P*values \\< 0.05 were considered statistically significant. ", "All analyses were performed using SPSS 15.0 (SPSS Inc, Chicago, IL, USA).", "\n\nResults\n=======\n\nPatient characteristics\n-----------------------\n\nBreast cancer patients with follow-up and *TP53*R72P and *MDM2*SNP309 genotypes from three hospital-based and one population-based study within the Breast Cancer Association Consortium were included for analysis (n = 3,749) (Table [1](#T1){ref-type=\"table\"}). ", "Frequencies of *TP53*R72P and *MDM2*SNP309 and clinicopathologic characteristics of the breast cancer patients in the four studies are shown in Table [2](#T2){ref-type=\"table\"}. ", "We have described and discussed earlier the small difference in *MDM2*SNP309 allele frequencies between European populations \\[[@B1]\\] while difference in patient characteristics between studies can mostly be attributed to differences in patient selection criteria (Table [1](#T1){ref-type=\"table\"}). ", "Mean follow-up was 7.7 years (SD 4). ", "A small number patients (n = 26) were carriers of the homozygous rare variants for both polymorphisms (Table [3](#T3){ref-type=\"table\"}).", "\n\n###### \n\nGerm-line variants and clinicopathologic characteristics of breast cancer patients by study\n\n -------------------------------------------------------------------------------------------------------------------------------------------------------------------\n ABCS\\ HABCS\\ HEBCS\\ SEARCH\\ *P value\\** \n N = 1076 N = 152 N = 599 N = 1922 \n --------------------------- ----------------------------- -------------- --------- -------------- ---------- -------------- ------ -------------- ------ ----------\n ***MDM2*SNP309** **TT** 444 41.3 55 36.2 183 30.6 774 40.3 \n\n **GT** 487 45.3 73 48.0 311 51.9 913 47.5 \n\n **GG** 145 13.5 24 15.8 105 17.5 235 12.2 \\< 0.001\n\n \n\n ***TP53*R72P** **GG** 570 53.0 85 55.9 314 52.4 1052 54.7 \n\n **GC** 422 39.2 55 36.2 236 39.4 733 38.1 \n\n **CC** 84 7.8 12 7.9 49 8.2 137 7.1 0.9\n\n \n\n **Stage** **1** 341 31.9 83 69.2 205 36.9 861 52.4 \n\n **2** 581 54.4 36 30.0 295 53.1 713 43.4 \n\n **3** 146 13.7 1 0.8 56 10.1 69 4.2 \\< 0.001\n\n **Missing** 8 32 43 279 \n\n \n\n **Differentiation grade** **1** 338 35.8 9 8.7 138 24.6 368 25.4 \n\n **2** 317 33.6 55 53.4 243 43.2 647 44.7 \n\n **3** 288 30.5 39 37.9 181 32.2 431 29.8 \\< 0.001\n\n **Missing** 133 49 37 476 \n\n \n\n **ER status tumor** **Negative** 240 34.2 18 15.4 135 23.2 175 19.8 \n\n **Positive** 461 65.8 99 84.6 446 76.8 708 80.2 \\< 0.001\n\n **Missing** 375 35 18 1039 \n\n \n\n **p53 status tumor** **Negative** 473 70.4 335 76.7 \n\n **Positive** 199 29.6 102 23.3 0.02\n\n **Missing** 404 152 162 1922 \n\n \n\n **Vital status patient** **Alive** 694 64.5 129 84.9 462 77.1 1596 83.0 \n\n **Deceased, all** 382 35.5 23 15.1 137 22.9 326 17.0 \\< 0.001\n\n **Deceased, breast cancer** 337 20 105 \n\n \n\n **Years of diagnosis** **Range** 1974 to 1994 1997 to 2003 1997 to 1998 1991 to 1996 \n\n \n\n **Age at diagnosis** **Mean ± SD** 42.8 5.2 56.8 11.3 56.4 12.8 50.1 7.7 \\< 0.001\n\n \n\n **Follow-up** **Mean ± SD** 10.5 5.7 6.5 1.9 7.3 2.1 6.3 2.1 \\< 0.001\n -------------------------------------------------------------------------------------------------------------------------------------------------------------------\n\n\\* *P*value of comparison of either categories of non-missing data among studies (by chi-square) or comparison of continuous data (by t-test).", "\n\n###### \n\nFrequencies of *TP53*R72P and *MDM2*SNP309 germ-line variants\n\n *TP53*R72P GG GC CC \n --------------------- ----- ------ ----- ------ ----- -----\n ***MDM2*SNP309 TT** 799 54.9 546 37.5 111 7.6\n **GT** 940 52.7 699 39.2 145 8.1\n **GG** 282 55.4 201 39.5 26 5.1\n\nBreast cancer survival by TP53 R72P, MDM2 SNP309 genotype, and p53 tumor status\n-------------------------------------------------------------------------------\n\nOverall survival of patients did not differ by carriership of either germ-line variant, R72P or SNP309, alone in the pooled analyses (Table [4](#T4){ref-type=\"table\"}). ", "Tumor p53 status was available for 1109 patients from the ABCS and HEBCS series (Table [1](#T1){ref-type=\"table\"}). ", "In both series, the patients with p53-positive tumors showed poorer overall survival than the patients with p53-negative tumors (pooled HR 1.5 (1.2-1.9), *P*= 0.002; Table [4](#T4){ref-type=\"table\"}).", "\n\n###### \n\nHR estimates of overall survival\\* by *TP53*R72P, *MDM2*SNP309 and p53\n\n -------------------------------------- -------- ---------------------------------- -------------- -------\n ***TP53*R72P\\*\\*** **HR** **Lower and upper limit 95% CI** ***P*value** \n\n \n\n **ABCS** \n\n  **GC** 0.99 0.79 1.26 0.95\n\n  **CC** 0.72 0.45 1.16 0.17\n\n **HABCS** \n\n  **GC** 1.40 0.60 3.45 0.46\n\n  **CC** 2.54 0.69 9.34 0.16\n\n **HEBCS** \n\n  **GC** 1.17 0.82 1.67 0.40\n\n  **CC** 1.72 1.00 2.98 0.05\n\n **SEARCH** \n\n  **GC** 1.18 0.95 1.49 0.14\n\n  **CC** 0.93 0.59 1.48 0.77\n\n **Pooled**^†^ \n\n  **GC** 1.11 0.96 1.28 0.18\n\n  **CC** 1.00 0.76 1.31 0.97\n\n \n\n ***MDM2*SNP309\\*\\*** \n\n \n\n **ABCS** \n\n  **TG** 0.93 0.73 1.18 0.54\n\n  **GG** 0.99 0.70 1.40 0.97\n\n **HABCS** \n\n  **TG** 0.60 0.25 1.46 0.26\n\n  **GG** 0.36 0.08 1.65 0.19\n\n **HEBCS** \n\n  **TG** 0.76 0.53 1.11 0.16\n\n  **GG** 0.91 0.56 1.47 0.69\n\n **SEARCH** \n\n  **TG** 1.03 0.82 1.31 0.78\n\n  **GG** 1.43 1.03 1.97 0.03\n\n **Pooled**^†^ \n\n  **TG** 0.93 0.80 1.08 0.34\n\n  **GG** 1.11 0.90 1.37 0.31\n\n \n\n **p53 status tumor \\*\\*** \n\n \n\n **ABCS** \n\n  **p53 positive** 1.31 0.96 1.80 0.09\n\n **HEBCS** \n\n  **p53 positive** 1.93 1.27 2.94 0.002\n\n **Pooled**^†^ \n\n  **p53 positive** 1.50 1.16 1.93 0.002\n\n **Breast cancer-specific survival**\\ \n **Pooled**^†^ \n\n  **p53 positive** 1.57 1.20 2.05 0.001\n -------------------------------------- -------- ---------------------------------- -------------- -------\n\n\\*Overall survival including all studies unless otherwise specified; \\*\\* HRs of heterozygous and homozygous rare allele groups have been calculated by comparison to the reference categories of common alleles: *TP53*R72P = GG; *MDM2*SNP309 = TT; p53 = negative tumors; ^†^Pooled analyses have been adjusted for study.", "\n\nDifferential effect of TP53 R72P on breast cancer survival stratified for p53 tumor status\n------------------------------------------------------------------------------------------\n\nIn the patient group with p53-negative tumors, the actuarial breast cancer-specific survival for the patients carrying the *TP53*CC genotype (Pro/Pro) was worse, though not statistically significantly, at 10 years of follow-up as compared to those carrying *TP53*GG/GC (Arg/Arg; Arg/Pro) (71% versus 80% *P*= 0.07; Figure [1](#F1){ref-type=\"fig\"}). ", "The interaction terms between p53 expression and *TP53*R72P were not significant in a multivariate Cox regression analysis, but considering the difference seen in the actuarial curves we still considered it useful to perform Cox analyses stratified for p53 expression. ", "Patients with the *TP53*CC genotype had worse breast-cancer specific survival (HR adjusted for study, age, stage, grade and ER: 1.79 (1.05 to 3.05, *P*= 0.03) (Table [5](#T5){ref-type=\"table\"}). ", "Results for overall survival were in line with those of breast-cancer specific survival, but did not reach statistical significance (*P*= 0.06, Table [5](#T5){ref-type=\"table\"}).", "\n\n{#F1}\n\n###### \n\nHR estimates of overall and breast cancer-specific survival by *TP53*R72P, in p53 negative and positive tumors (multivariate models)\n\n ------------------------------------- -------- ---------------------------------- -------------- ------\n ***TP53*R72P** **HR** **Lower and upper limit 95% CI** ***P*value** \n \n **Overall survival** \n **p53 negative tumors** \n  **GC** 1.11 0.80 1.52 0.54\n  **CC** 1.63 0.97 2.74 0.06\n **p53 positive tumors** \n  **GC** 0.82 0.53 1.27 0.37\n  **CC** 0.86 0.34 2.18 0.75\n \n **Breast cancer-specific survival** \n **p53 negative tumors** \n  **GC** 0.97 0.68 1.37 0.85\n  **CC** 1.79 1.05 3.05 0.03\n **p53 positive tumors** \n  **GC** 0.90 0.57 1.42 0.65\n  **CC** 1.00 0.39 2.55 1.00\n \n ***MDM2 SNP309*** **HR** **Lower and upper limit 95% CI** ***P*value** \n \n **Overall survival** \n **p53 negative tumors** \n  **TG** 0.84 0.60 1.17 0.30\n  **GG** 1.28 0.85 1.94 0.24\n **p53 positive tumors** \n  **TG** 0.78 0.49 1.24 0.30\n  **GG** 0.78 0.41 1.48 0.45\n \n **Breast cancer-specific survival** \n **p53 negative tumors** \n  **TG** 0.92 0.64 1.31 0.63\n  **GG** 1.41 0.90 2.19 0.13\n **p53 positive tumors** \n  **TG** 0.76 0.47 1.23 0.27\n  **GG** 0.69 0.35 1.39 0.30\n ------------------------------------- -------- ---------------------------------- -------------- ------\n\nPooled analyses for studies with p53 information (ABCS and HEBCS); HRs of heterozygous and homozygous rare allele groups have been calculated by comparison to the reference categories of the homozygous common allele: *TP53*R72P = GG, and *MDM2*SNP309 = TT; analyses have been adjusted for study, age, stage, grade and ER.", "\n\nWithin the patient group with p53-positive tumors, breast cancer-specific survival stratified by *TP53*R72P seemed to show inconsistent results between studies though none were significant, that is, per allele HR (adjusted for age, stage, grade and ER) in the ABCS study was 0.74 (0.45 to 1.22) and in the HEBCS study 1.46 (0.79 to 2.69). ", "The pooled HR (adjusted for study, age, stage, grade and ER) was 0.82 (0.53 to 1.27) for heterozygous and 0.86 (0.34 to 2.18) for homozygous C-allele carriers (Table [5](#T5){ref-type=\"table\"}). ", "There was no evidence for a differential effect of *MDM2*SNP309 by p53 tumor status on survival (Table [5](#T5){ref-type=\"table\"}).", "\n\nCombined effects of TP53 R72P and MDM2 SNP309 on breast cancer survival\n-----------------------------------------------------------------------\n\n*MDM2*SNP309 showed a differential actuarial overall survival stratified by *TP53*R72P in the pooled analyses (n = 3,749), that is, homozygous carriers of the G-allele in *MDM2*had worse survival within the group of *TP53*GC carriers (GG: 65% versus GT: 72% and TT: 76%, *P*= 0.006; Figure [2](#F2){ref-type=\"fig\"}). ", "The same trend was visible in the *TP53*homozygous CC group (n = 26 GG/CC), but this was not statistically significant. ", "Within the *TP53*C-allele carriers combined, *MDM2*GG carriers had significantly worse survival compared to TT/TG carriers: 64% versus 75%, *P*= 0.001. ", "In multivariate analyses (adjusting for study, age, stage, grade and ER) the interaction term for *TP53*GC and *MDM2*GG was significant (*P*= 0.028), also if additional interaction terms for *TP53*R72P and p53 expression were included (*P*= 0.027). ", "The multivariate models (adjusting for study, age, stage, grade and ER) stratified for *TP53*R72P (analogue to Figure [2](#F2){ref-type=\"fig\"}) showed that *MDM2*GG carriers had significantly worse survival compared with *MDM2*TT carriers only within the *TP53*C-allele carriers; more specifically, within *TP53*CG carriers: HR 1.43 (1.05 to 1.96), *P*= 0.02; within *TP53*CC carriers HR 1.39 (0.56 to 3.48), *P*= 0.48 (Table [6](#T6){ref-type=\"table\"}); within *TP53*CG and CC carriers combined: HR (adjusted for study, age, stage, grade and ER) 1.46 (1.09 to 1.96), *P*= 0.01.", "\n\n{#F2}\n\n###### \n\nHR estimates of multivariate analyses for *MDM2*SNP309 stratified by *TP53*R72P\n\n --------------------- -------------------- ---------------------------------- -------------------- --------- ---------------------------------- -------------- -------- ---------------------------------- -------------- ------ ------- ---------\n **HR** **Lower and upper limit 95% CI** ***P*value** **HR** **Lower and upper limit 95% CI** ***P*value** **HR** **Lower and upper limit 95% CI** ***P*value** \n \n ***TP53*R72P: GG** ***TP53*R72P: GC** ***TP53*R72P: CC** \n \n ***MDM2*SNP309 TT** 1.0 (Ref) 1.0 (Ref) 1.0 (Ref) \n  **TG** .90 .73 1.11 .33 .94 .73 1.20 .61 .85 .47 1.55 .60\n  **GG** .85 .63 1.16 .31 1.43 1.05 1.96 .02 1.39 .56 3.48 .48\n \n **p53 negative** 1.0 (Ref) 1.0 (Ref) 1.0 (Ref) \n  **positive** 1.37 .96 1.96 .08 .89 .58 1.35 .57 .76 .25 2.30 .63\n  **missing** 1.63 1.16 2.28 .005 1.66 1.13 2.44 .009 .53 .21 1.34 .18\n \n **Stage 1** 1.0 (Ref) 1.0 (Ref) 1.0 (Ref) \n  **2** 2.51 1.90 3.33 \\< .001 2.55 1.88 3.46 \\< .001 1.48 .73 2.97 .28\n  **3** 7.10 5.05 9.97 \\< .001 7.56 5.18 11.02 \\< .001 5.49 2.17 13.87 \\< .001\n  **missing** 3.69 2.42 5.63 \\< .001 2.55 1.54 4.21 \\< .001 1.60 .46 5.57 .46\n \n **Grade 1** 1.0 (Ref) 1.0 (Ref) 1.0 (Ref) \n  **2** 1.80 1.27 2.54 .001 1.12 .77 1.62 .55 3.28 1.29 8.35 .01\n  **3** 2.63 1.84 3.76 \\< .001 2.39 1.65 3.46 \\< .001 4.08 1.55 10.79 .005\n  **missing** 1.84 1.21 2.78 .004 1.391 .87 2.22 .17 2.11 .64 6.92 .22\n \n **ER negative** 1.0 (Ref) 1.0 (Ref) 1.0 (Ref) \n  **positive** .63 .48 .84 .002 .61 .44 .83 .002 .59 .27 1.26 .17\n  **missing** .65 .46 .91 .01 .58 .40 .84 .003 .90 .36 2.25 .82\n \n **Age** 1.02 1.01 1.03 .001 1.00 .99 1.02 .54 1.01 .98 1.04 .70\n --------------------- -------------------- ---------------------------------- -------------------- --------- ---------------------------------- -------------- -------- ---------------------------------- -------------- ------ ------- ---------\n\nModels have also been adjusted for study.", "\n\nDiscussion\n==========\n\nIn the survival analyses including 3,749 breast cancer patients from Finland, The Netherlands, Germany and United Kingdom, we showed combined effects of two germ-line polymorphisms, *TP53*R72P, *MDM2*SNP309, and p53 tumor expression (by immunohistochemistry). ", "Firstly, we confirmed our earlier observation in Finnish patients \\[[@B10]\\] that *TP53*R72P homozygous carriership predicts a worse survival in patients with p53-negative tumors, also when adjusted for clinical prognostic variables. ", "Thus, in the absence of inactivating p53 mutations in the tumor, the 72P variant form of p53 protein may have a compromising effect on the p53 apoptotic function, leading to reduced survival of the patients. ", "Similarly, a study of 414 Chinese breast cancer patients reported that the 72P homozygous (CC) genotype was associated with both poorer five-year overall survival (five to eight percentile difference, *P*= 0.04) and poorer disease-free survival among the patients with a wild-type p53 in their tumors (n = 346) \\[[@B16]\\]. ", "In line with other studies published we did not observe an effect of carriership of R72P alone on survival of patients \\[[@B12]-[@B16]\\].", "\n\nNo significant difference in survival by *TP53*R72P carriership was observed among the patients with p53-positive tumors, who showed a worse survival overall compared to p53-negative tumors. ", "In the pooled analysis, CC homozygote patients with p53-positive tumors even tended to have a better survival. ", "In the study by Xu et al. ", "in Chinese breast cancer patients \\[[@B16]\\], the CC homozygote patients also had non-significant better survival than the GG homozygotes and heterozygotes within the group of patients with p53-mutated tumors.", "\n\nThe finding of CC homozygote (72P) carriers having poorer survival is consistent with the R72 variant of wild-type p53 being a more potent inducer of apoptosis than the wild-type 72P variant. ", "It has been suggested that R72 homozygotes may respond more favorably to radiation or chemotherapy \\[[@B27]\\]. ", "Response rate after chemo-radiotherapy of advanced squamous cell carcinomas of head and neck and survival was higher in patients with the R72 allele compared to those with the 72P allele \\[[@B28]\\]. ", "These favorable effects of the R72 allele may, however, be reversed by a somatic p53 mutation on this allele, as has been reported in squamous cell carcinomas of head and neck \\[[@B29],[@B30]\\]. ", "In line with this, retention of the R72 allele with loss of the 72P allele in the tumor tissue has been associated with reduced survival in heterozygous breast cancer patients \\[[@B31]\\].", "\n\nCarriership of *MDM2*SNP309 alone did not affect survival of patients in our study and two other, smaller studies \\[[@B13],[@B22]\\]. ", "However, we found an 11 percentile survival difference for homozygous *MDM2*G-allele carriers within the group of *TP53*C-allele (72P) carriers. ", "Biologically this seems plausible considering the reduced apoptotic function of the *TP53*Pro-variant \\[[@B6],[@B7]\\] and the attenuation of the p53 pathway by mdm2, the production of which is increased by the SNP309 G-variant \\[[@B17]\\]. ", "In addition, the interaction of both polymorphisms remained statistically significant in multivariate models adjusting for clinical prognostic factors.", "\n\nWe did not observe evidence for a combined effect of SNP309 and p53 tumor expression (as shown here by results of SNP309 stratified by p53 status in Table [5](#T5){ref-type=\"table\"}, but obviously p53 did also not have a differential effect on survival stratified by SNP309). ", "This is in contrast to a previous, smaller study (n = 248) in the American population, which suggested that tumor p53 status was associated with breast cancer survival only among patients homozygous for the *MDM2*SNP309 T-allele and not among carriers of the variant G-allele \\[[@B22]\\]. ", "Though our study is one of the largest published studies on combined effects of the germline genetic variation and tumor somatic events, the numbers are still small for looking at such modifying effects on survival.", "\n\nMany studies have confirmed that mutated p53 is a prognostic factor in breast cancer. ", "The risk of dying of breast cancer for patients with a p53 mutation in their tumor has been estimated to be two to five-fold compared to patients with wild-type p53 tumors \\[[@B32],[@B33]\\]. ", "Positive immunostaining for p53 is in general considered to indicate somatic p53 mutation and an impaired p53 pathway, though the correlation with *TP53*mutations is incomplete \\[[@B34],[@B35]\\]. ", "The accumulation of p53 in the tumors detected by immunohistochemistry was a prognostic marker of poorer survival in both our series with p53 immunohistochemistry data available (the HEBCS and ABCS series). ", "This effect was somewhat stronger in the HEBCS series, which may be explained by the more stringent cut-off used (20% positive tumor cells compared to 10% in the ABCS series).", "\n\nConclusions\n===========\n\nWe have shown here that *TP53*R72P may have additional prognostic value especially among patients with p53-negative tumors. ", "However, the effect of p53 on outcome may be influenced by adjuvant systemic therapy (for example, \\[[@B31],[@B36]\\], reviewed in Bertheau \\[[@B37]\\]) and larger studies will be needed to address this question. ", "Our study is one of the few that have shown an interaction of germ-line variants, that is, *TP53*R72P and *MDM2*SNP309, in breast cancer survival. ", "The results, showing a statistically significant interaction of the p53 Pro-variant and the GG genotype of *MDM2*SNP309, are in line with our a priori biologically-supported hypothesis, which is, the role of enhanced DNA repair function of the Pro-variant, combined with increased expression of the Mdm2 protein, and thus overall attenuation of the p53 pathway in the tumor cells. ", "These results suggest that even subtle differences in p53 apoptotic function caused by synergistic polymorphisms may affect patient\\'s survival, possibly by modifying treatment response. ", "Altogether, our findings are in line with biological evidence in literature, and in the future, may have also clinical significance for models of breast cancer prognosis or treatment. ", "However, because this is the first report on the combined effect of *TP53*R72P and *MDM2*SNP309 on breast cancer survival and we cannot exclude a chance finding, other studies to confirm this will be necessary. ", "Larger studies will be needed also to investigate the effect of specific treatment modalities on the survival by *TP53*R72P and *MDM2*SNP309.", "\n\nAbbreviations\n=============\n\nABCS: Amsterdam Breast Cancer Study; ER: estrogen receptor; HABCS: Hannover Breast Cancer Study; HEBCS: Helsinki Breast Cancer Study; HR: hazard ratio; SD: standard deviation; SEARCH: Studies of Epidemiology and Risk Factors in Cancer Heredity; TMA: tissue micro array.", "\n\nCompeting interests\n===================\n\nThe authors declare that they have no competing interests.", "\n\nAuthors\\' contributions\n=======================\n\nMKS, TD and HN took final responsibility for the decision to submit the paper for publication; all other authors read and approved the manuscript. ", "MKS, JT, FEVL, LJVTV, PDPP, DFE, TD, CB, HN were responsible for the study design. ", "MKS, JT, AB, MS, MH, TD, SAR, RF were responsible for data acquisition and collection. ", "MKS and JT did the data analyses. ", "Data interpretation was carried out by MKS, JT, AB, RF, CB, and HN. ", "MKS, AB, TD, CB, and HN wrote the paper. ", "All authors read and approved the manuscript.", "\n\nAcknowledgements\n================\n\nWe wish to thank Dr Hannaleena Eerola and Dr Kirsimari Aaltonen and Nina Puolakka RN for their help with the Finnish patient data. ", "The Finnish Cancer registry is gratefully acknowledged for the cancer data. ", "The Helsinki study has been financially supported by the Helsinki University Central Hospital Research Fund, Academy of Finland (110663), Finnish Cancer Society, the Sigrid Juselius Foundation, and the Ida Montin Foundation. ", "We highly appreciate the contributions of our Dutch colleagues at the NKI-AVL, among others Renate Udo, Linde Braaf and Hans Peterse^†^; and at LUMC: Rob Tollenaar, Vincent Smit and Cees Cornelisse; the Dutch study was financed by the Dutch Cancer Society (NKI 2001-2423; 2007-3839) and the Cancer Genomics Center (Dutch Genomics Initiative). ", "We are grateful to our German colleagues Michael Bremer, Andreas Meyer, Johann H. Karstens and Peter Hillemanns for their support of the study at Hannover Medical School. ", "The SEARCH study is funded by Cancer Research UK, and DFE is a Principal Research Fellow and PDP is a Senior Clinical Research Fellow of CRUK.", "\n" ]

{ "pile_set_name": "PubMed Central" }

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[ "Max Planck Inst. ", "for Biophysical Chemistry\n\nEver since the seventeenth century, when the early microbiologist Antonie van Leeuwenhoek focused light through lenses and marvelled at the cells that swam before his eyes, microscopes have opened up new vistas of discovery. ", "This year, the Nobel Prize in Chemistry went to three scientists who defied the limits of light microscopes to reveal sharp images of molecular-scale structures in living cells.", "\n\nThe advances made by Stefan Hell, William Moerner and Eric Betzig in the 1990s and 2000s mean that biologists can now see, in real time, how proteins are distributed and move inside cells — at the junctions between neurons, for example, or in fertilized eggs dividing into embryos.", "\n\n“It is really a revolution for the life sciences, because we can see structures that we could never see before,” says Stefan Jakobs, who works with super-resolution techniques at the Max Planck Institute for Biophysical Chemistry in Göttingen. ", "Or as the Nobel committee put it: “Microscopy has become nanoscopy.”", "\n\nNo matter how clean their lenses, optical microscopes inevitably provide a blurry view of the molecules inside cells, as German physicist Ernst Abbe realized in 1873. ", "The laws of physics dictate that visible light cannot distinguish between objects closer to each other than around 200 nanometres (around half the wavelength of visible light) — they will appear as one blob. ", "Such resolution, known as Abbe's diffraction limit, is good enough to reveal the organelles inside cells but not to see their detailed structures. ", "Microscopes that use beams of electrons, rather than light, have finer resolution, but they can be used only in a vacuum, limiting their use to dead tissue.", "\n\nAbbe's limit cannot be overcome, but the 2014 Nobel prizewinners pioneered ways to work around it using fluorophores, or fluorescent molecules. ", "Now routinely used in biological imaging, fluorophores emit light when hit by lasers of a certain wavelength.", "\n\nBeating the blur\n\nL.A. Cicero/Stanford Univ.", "\n\nIn 1989, William Moerner, now at Stanford University in California, but then at the IBM Almaden Research Center in San Jose, detected the faint fluorescence of a single molecule. ", "In 1997, while working at the University of California in San Diego, he found a way to control the fluorescence and switch the molecules on and off like lamps. ", "Still, these single molecules could be distinguished only if they were more than 200 nanometres apart.", "\n\nTwo years earlier, Eric Betzig, who was at that time working at Bell Labs in Murray Hill, New Jersey, had proposed that if different molecules inside a cell could be made to glow with different colours, researchers should be able to increase the resolution by taking a series of snapshots — first the red molecules, then the green, then the blue. ", "Any fluorophores of the same colour would have to be more than 200 nanometres apart, but the superimposed images would produce a much finer-resolution structure. ", "Moerner went on to show that identical molecules could be made to fluoresce at different times, a discovery that ultimately made Betzig’s vision a reality.", "\n\nMatt Staley/Howard Hughes Medical Inst.", "\n\nIt would take a decade before Betzig demonstrated his idea in practice. ", "He left academic science to work in his father’s machine tool business in Michigan. ", "In 2006, working at the Howard Hughes Medical Institute’s Janelia Farm research campus in Ashburn, Virginia, he used the technique to take a super-resolution picture of a lysosome protein dotted with green fluorescent molecules as labels. ", "The technique can now get down to a resolution of 20 nanometres, says Markus Sauer, who studies super-resolution microscopy at the University of Würzburg, Germany.", "\n\nMeanwhile Stefan Hell, working at the University of Turku in Finland, had found a way around Abbe's limit by a different technique, which also relies on switching fluorescent molecules on and off. ", "In 1994 he proposed using one laser to make a cluster of dye molecules fluoresce, and a second beam, of a different wavelength, to switch some of those fluorophores off — through a process described by Einstein in 1917.", "\n\nHell's trick is to use the second beam to outline the cluster illuminated by the first, so that only the molecules in a very narrow spot fluoresce. ", "The final image remains blurred, as light still cannot beat Abbe’s limit, but it is clear that light can have come only from the narrow central spot defined by the second beam, enabling researchers to pinpoint the light source.", "\n\nBuilding up a series of these tiny fluorescent spots creates a fine-resolution picture. ", "In theory, the central spot can be made as small as a few nanometres across, but in living cells, the limit is around 30 nanometres, Sauer says, because it is at this stage that fluorophores are usually destroyed by the intensity of the second beam.", "\n\nCellular world\n\nBernd Shuller/Max Planck Inst. ", "for Biophysical Chemistry\n\n“It was my view, at least, that so much physics happened in the twentieth century that it was impossible there was no phenomenon that would allow you to overcome the diffraction barrier,” Hell, who now works at the Max Planck Institute for Biophysical Chemistry in Göttingen, told the Nobel committee, on learning that he had won the prize.", "\n\nIndeed, every quantum-physics principle that Hell used had been discovered by physicists by the end of the 1920s, notes Thomas Klar, head of the Institute of Applied Physics at Johannes Kepler University in Linz, Austria, who co-authored the proof-of-principle paper with Hell in 2000.", "\n\nThe call from the Nobel committee caught Hell while he was reading a scientific paper. ", "Afterwards, he said, “I read the paragraph that I wanted to read to the end, then I called up my wife and tried to reach some of the people who are close to me.\"", "\n\nThe techniques devised by this year's prizewinners are not yet routine but are used by many biologists to take remarkable pictures of structures inside cells. ", "Hell has published videos of vesicles 40 nanometres across moving in a nerve cell. ", "Xiaowei Zhuang, a chemist at Harvard University in Cambridge, Massachusetts, has invented her own variation, called stochastic optical reconstruction microscopy, and used it to show how actin filaments wrap around the circumference of axons in a ring-like pattern. \"", "There will be many new versions of super-resolution microscopes in future,\" Hell says.", "\n\n" ]

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[ "Introduction {#s1}\n============\n\nA violent and unprovoked attack by one person on another unfolds in close view of an unrelated bystander: under what conditions will the bystander be likely to intervene to help the victim? ", "In this paper we address the hypothesis that group affiliation between the bystander and the victim provides a powerful incentive for the bystander to try to intervene to stop the attack, or prevent harm to the victim, and in particular that this operates even though the perpetrator and victim are virtual human characters. ", "Our experiment involved fans of an English football team, Arsenal. ", "In one experimental condition (in-group) the fan conversed with a virtual character that was clearly an Arsenal supporter and in another condition the character was just a general football enthusiast but not an Arsenal fan (out-group). ", "The virtual character was later threatened by a perpetrator that, in the in-group condition, specifically attacked his Arsenal affiliation. ", "Our expectation was that based on group affiliation, those in the in-group would intervene more than those in the out-group. ", "First we place this in the general context of studies of bystander intervention, and then describe the detailed design of the experiment and the results.", "\n\nResearch on the behaviour of bystanders in emergencies began with the response to the rape and murder of Kitty Genovese in New York in 1964. ", "Social psychologists Bibb Latane and John Darley read a report on the murder in the New York Times suggesting that 38 witnesses had watched the murder unfold over 30 minutes from their apartment windows-- and yet failed to intervene. ", "In order to understand why this might have happened, they set out to create laboratory based experimental analogies of the event. ", "They set up carefully choreographed situations in which bystanders were faced with a non-violent emergency situation while on their own or in the presence of others [@pone.0052766-Darley1], [@pone.0052766-Latan1]. ", "The research led to the discovery of the 'bystander effect' -- the idea that people are more likely to intervene on their own than in the presence of others [@pone.0052766-Darley1]. ", "This is one of the most reliable and robust findings in social psychology [@pone.0052766-Latan2], [@pone.0052766-Fischer1].", "\n\nHowever, as Cherry pointed out [@pone.0052766-Cherry1], through translating the events surrounding the Genovese murder into laboratory settings, Latane and Darley neglected some of the key features of the event. ", "Despite the fact that the original murder involved violence by a man against a woman, subsequent experimental analogies tended to remove both the gendered nature of the attack and the violence. ", "Although there are thousands of studies using non-violent emergency settings, it is possible to find only a few experiments that did retain violence as the emergency variable [@pone.0052766-Harari1], [@pone.0052766-Schwartz1], [@pone.0052766-Schwartz2], [@pone.0052766-Shotland1], [@pone.0052766-Borofsky1]. ", "These found results that were at odds with the traditional bystander paradigm. ", "In violent emergencies, what seemed to be most important about the likelihood of bystander intervention was not the presence of others, but rather the bystanders' beliefs about the nature of the relationship between perpetrator and victim [@pone.0052766-Shotland1], [@pone.0052766-Borofsky1]. ", "In an experiment that did vary the number of bystanders to a violent emergency Harari et al. [", "@pone.0052766-Harari1] showed that the presence of others actually enhanced the likelihood of bystander intervention in a simulated rape situation. ", "This finding has been supported by contemporary work which presents violence to participants by means of a CCTV video link, where the presence of others is not found to inhibit helping [@pone.0052766-Fischer2] and can sometimes enhance it [@pone.0052766-Levine1]. ", "A recent meta-analysis by Fisher and colleagues [@pone.0052766-Fischer1] confirms that intervention behaviour in violent emergencies does not fit the traditional bystander effect explanation.", "\n\nIf violent emergencies are different in some way, it is important to understand the processes at work. ", "Almost all violence research shares a similar limitation. ", "In order to circumvent the practical and ethical problems of presenting violence in experimental settings, these experiments tend to avoid placing participants in *direct* contact with the violence itself. ", "The only exception is the work described in [@pone.0052766-Borofsky1] in which a role-play setting was used, and confederates actually staged a violent confrontation in front of naive participants who were also taking part in the role-play game. ", "However, it is highly unlikely that contemporary ethics boards would allow this kind of design. ", "The other studies either have the violence happening at a distance where it is possible to avoid the event [@pone.0052766-Harari1] or present the violence as happening contemporaneously but where it can only be heard [@pone.0052766-Schwartz1], [@pone.0052766-Schwartz2], [@pone.0052766-Shotland1], or happening in another room where it can be seen on CCTV link [@pone.0052766-Fischer2]. ", "This distancing of participants from the violence is required to satisfy the ethical and practical difficulties of experimental design, but may itself introduce psychological effects that interfere with the veridical nature of the situation. ", "Imagining the violence, or having it happen in another room, is not the same as being physically where the violence erupts.", "\n\nIn [@pone.0052766-Rovira1] we argued that the use of immersive virtual environments (IVE) goes some way towards solving this problem, since there is mounting evidence that when people are faced with events and situations in an IVE they tend to behave and respond as if these were real [@pone.0052766-SanchezVives1]. ", "IVEs portray a simulated computer generated reality at life size that is sensorially surrounding. ", "Participants perceive this world through wide field-of-view stereo vision and sound. ", "The form of perception involves more or less natural sensorimotor contingencies - meaning that the whole body is used for perception much as in physical reality, based at least on head-gaze direction and orientation achieved through head-tracking. ", "This gives rise to the sensation of being in the virtual place that is depicted, a place-illusion. ", "Additionally when there are dynamically unfolding events in the environment that personally refer to the participant, and where actions of the participant apparently cause responses in the virtual environment, this gives rise to a plausibility-illusion, meaning that events have the illusory quality of being real. ", "When the participant has the double illusion - of being in the virtual place and where events that are happening are apparently really happening, this can give rise to behaviour and responses that are appropriate to the situation as if it were playing out in reality [@pone.0052766-Slater1].", "\n\nIVEs provide therefore a powerful tool for experimental studies in social psychology [@pone.0052766-Blascovich1] and classic effects such as proxemics [@pone.0052766-Hall1] where distances that people maintain between themselves are governed by social norms, have been reproduced several times in IVEs with respect to virtual humanoid characters [@pone.0052766-Wilcox1], [@pone.0052766-Llobera1], [@pone.0052766-Bailenson1]. ", "Moreover, IVEs have been useful for experiments that would otherwise be difficult to carry out in any other way, such as the study of male risk taking in the presence of observers, specifically the differential effects of the observers being male or female [@pone.0052766-Frankenhuis1].", "\n\nCloser to the present study which focuses on responses to violence, the Stanley Milgram obedience paradigm [@pone.0052766-Milgram1] has been reproduced with IVE avoiding the ethical difficulties of deception [@pone.0052766-Slater2], [@pone.0052766-Cheetham1]. ", "IVEs provide environments completely under control of a computer program but where people respond realistically. ", "Every experimental condition can be exactly reproduced across trials as needed, and hence can be used for laboratory based experiments.", "\n\nIt has been argued before that IVEs provide an excellent tool for the study of prosocial behaviour [@pone.0052766-Gillath1]. ", "The experiment described in the present study is specifically concerned with the likelihood of prosocial behaviour when participants are placed in direct proximity to violent behaviour. ", "We explore the hypothesis that the psychological relationships between bystanders and the others involved are important in bystander behaviour, in this case specifically the relationship between the bystander and the victim [@pone.0052766-Levine1], [@pone.0052766-Levine2], [@pone.0052766-Levine3], [@pone.0052766-Levine4]. ", "The experimental conditions provide a context where it is certain that the violence between perpetrator and victim is of the same magnitude and intensity for each experimental trial. ", "Participants (n = 40) *all supporters of the Arsenal Football Club*, entered into a virtual reality that represents a bar. ", "A male virtual human (V) approached and conversed with them about football for a few minutes. ", "In one condition V wore an Arsenal football shirt and spoke enthusiastically about the club (in-group condition). ", "In a second condition V wore an unaffiliated red sports shirt, and asked questions about Arsenal without special enthusiasm, using neutral responses and displaying ambivalence about Arsenal's prospects (out-group condition). ", "After a few minutes of this conversation another male virtual human (P, perpetrator) who had been sitting by the bar walked over to V (victim) and started an argument that he continually escalated until it became a physical attack ([Figure 1](#pone-0052766-g001){ref-type=\"fig\"}).", "\n\n{#pone-0052766-g001}\n\nThe main response variable was the extent to which the participant attempted to intervene during this confrontation. ", "Interventions were verbal utterances or physical moves towards the two virtual characters and were coded from video recordings by two independent researchers (Methods). ", "There were two binary factors *group* and *LookAt*. *", "Group* was whether V was in-group (Arsenal supporter) or out-group with respect to the participant. *", "LookAt* was whether or not occasionally during the confrontation V would look towards the participant or not (*LookAt* = 'on' or 'off'). ", "The experiment used a between-groups design, with n = 40, 10 participants allocated arbitrarily to one of the four cells of the 2×2 design. ", "The degree of support for the Arsenal club was similar between the 4 experimental conditions ([Text S1](#pone.0052766.s004){ref-type=\"supplementary-material\"}). ", "At the end of their session they answered a questionnaire, and this was followed by an interview and debriefing. ", "The data from two participants could not be used due to video recording failures.", "\n\nResults {#s2}\n=======\n\nNumbers of Interventions {#s2a}\n------------------------\n\n[Table 1](#pone-0052766-t001){ref-type=\"table\"} shows the means and standard errors of the numbers of interventions indicating that the mean number of interventions was higher for the in-group than the out-group, but that the *LookAt* factor had no effect. ", "Two-way analysis of variance was carried out on the response variables, the number of physical (*nPhys*) and number of verbal (*nVerbal*) interventions. ", "ANOVA for *nPhys* indicates that the mean is greater for the in-group than for the out-group condition (P = 0.02) but with no significant differences for the *LookAt* factor and no interaction effect. ", "However, the residual errors of the fit were strongly non-normal (Shapiro-Wilk test P = 0.0008). ", "To overcome this problem a square root transformation was applied to *nPhys*. ", "This resulted in the same conclusions for *group* (P = 0.016, partial η^2^ = 0.15) and no significance for *LookAt* (P = 0.297, partial η^2^ = 0.03). ", "The normality of the residuals is improved although not ideal (Shapiro-Wilk P = 0.034). ", "For the response variable *nVerbal* the results were similar: ANOVA of *nVerbal* on *group* and *LookAt* shows no significant interaction term, *group* has significance level P = 0.095, and for *LookAt* P = 0.228. ", "However, again the residual errors are far from normal (SW P = 0.0008). ", "The square root transformation gives P = 0.060, partial η^2^ = 0.10 for *group* and P = 0.112, partial η^2^ = 0.07 for *LookAt*. ", "The residual errors are compatible with normality (SW P = 0.24).", "\n\n10.1371/journal.pone.0052766.t001\n\n###### Means and Standard Errors of Numbers of Interventions.", "\n\n![](", "pone.0052766.t001){#pone-0052766-t001-1}\n\n No. ", "Verbal Interventions \n ---------- -------------------------------- --------- ---------\n Outgroup 3.9±1.4 2.0±1.3 2.9±1.0\n Ingroup 6.8±1.8 4.7±1.9 5.8±1.3\n All 5.4±1.2 3.4±1.2 4.4±0.8\n **No. ", "Physical Interventions** \n Outgroup 2.8±1.1 1.8±1.0 2.3±0.7\n Ingroup 6.8±2.1 6.1±2.2 6.5±1.5\n All 4.9±1.3 4.1±1.3 4.5±0.9\n\nn = 9 for each of the two Out-group cells, n = 10 for each of the two In-group cells, n = 38 in total.", "\n\nThe factor *LookAt* represents whether the V avatar was *programmed* to occasionally look toward the participants. ", "Additionally, the post experience questionnaire included the statement (*VictimLooked*) \"After the argument started, the victim looked at me wanting help\" which was scored on a scale from 1 (least agreement) to 7 (most agreement). *", "VictimLooked* therefore represents the *belief* of the participants as to whether the victim *looked towards them for help*. ", "There is no significant difference between the mean *VictimLooked* score of those who were in the group *LookAt* = 'on' (mean 3.3, SD = 1.8, n = 20) and those in the group *LookAt* = 'off' (mean 4.0, SD = 1.5, n = 20) (P = 0.12, Mann-Whitney U). ", "Hence the response to this question was not based on the number of actual looks of the victim towards the participant, and therefore was a belief. ", "It turns out that *VictimLooked* plays a significant role in the number of interventions.", "\n\n[Figure 2](#pone-0052766-g002){ref-type=\"fig\"} shows the scatter plots of *nPhys* and *nVerbal* on the questionnaire response *VictimLooked* for the out-group and in-group. ", "These reveal a quite different relationship in the two cases. ", "In the case of the in-group there is a positive association between the number of interventions (verbal or physical) and the perception that the victim was looking towards the participant for help. ", "In the case of the out-group there appears to be no relationship in the *nPhys* case and a possible negative relationship in the *nVerbal* case. ", "Using the same strategy as above in order to obtain residual errors compatible with normality, ANCOVA of *nPhys* ^0.5^ on *group* with *VictimLooked* as a covariate shows that the slopes of the regression line are different between the in-group and out-group (P = 0.004, partial η^2^ = 0.22 for the slopes, SW P = 0.18). ", "For the number of verbal interventions, using *nVerbal* ^0.5^ the difference in slopes between in-group and out-group is significant at P = 0.004 (partial η^2^ = 0.22 for the slope, SW P = 0.12).", "\n\n{#pone-0052766-g002}\n\nThese results indicate that the response to the belief that the victim was looking towards the bystander for help was different between the in-group and out-group. ", "For those in the in-group condition the greater their belief that the victim was looking to them for help the greater the number of verbal and physical interventions. ", "For those in the out-group condition there is no such association. ", "These results are further corroborated using multivariate analysis of variance on the response vector (*nPhys* ^0.5^, *nVerbal* ^0.5^) ([Text S2](#pone.0052766.s005){ref-type=\"supplementary-material\"}).", "\n\nNumbers of Interventions - Symbolic Regression {#s2b}\n----------------------------------------------\n\nThe previous section provided standard analyses for these types of data. ", "Even though this revealed positive results consistent with our initial hypothesis, in this section we also employ a quite different method using symbolic regression, to throw further light on the experimental results. ", "The purpose is to consider the relationship between the number of interventions, and the experimental factors, but now also including any possible influence of the subjective variables as elicited through the post-experience questionnaire ([Table 2](#pone-0052766-t002){ref-type=\"table\"}). ", "Standard statistical analysis is based, amongst other things, on the assumption of linearity in the parameters. ", "But in such a complex situation as the one under consideration, on what grounds is such an assumption valid when considering the multivariate influence of a number of factors potentially influencing bystander intervention? ", "Symbolic regression does not rely on such linearity, being a method for discovering relationships between variables using the technique of genetic programming [@pone.0052766-Koza1] ([Text S3](#pone.0052766.s006){ref-type=\"supplementary-material\"}). ", "It has recently been shown to be able to discover complex physical laws automatically [@pone.0052766-Schmidt1], using a program called Eureqa, which was used in the analysis presented below. ", "In the context that we apply this technique here, we consider it as a data reduction method. ", "It allows us to succinctly represent the original data but with quite simple equations while preserving the variance in the original data. ", "It is not a technique that can be compared with statistical significance testing, it is rather a data exploration method, that can lead to understanding of complex data, where models generated by this technique can be used for hypothesis formation in later experimental study.", "\n\n10.1371/journal.pone.0052766.t002\n\n###### The Post-Questionnaire and Corresponding Variable Names.", "\n\n![](", "pone.0052766.t002){#pone-0052766-t002-2}\n\n Variable Statement\n --------------- --------------------------------------------------------------------------------------------------------------------\n Uncomfortable After the argument started, I was feeling uncomfortable with the situation.", "\n OtherSafety After the argument started I was sometimes concerned for the safety of the man being threatened.", "\n OwnSafety After the argument started I was sometimes concerned for my own safety.", "\n HelpMe After the argument started I looked around for help.", "\n OtherPeople After the argument started I looked around to check in case other people might arrive to make the situation worse.", "\n VictimLooked After the argument started, the victim looked at me wanting help.", "\n MoveAway After the argument started I felt I should move away from those people.", "\n AgressorAware After the argument started, the aggressor was aware of me looking at him.", "\n ShouldStopIt After the argument started, I felt I should do something to stop it.", "\n CouldStopIt After the argument started, I felt I could do something to stop it.", "\n GetOut After the argument started I felt that I needed to get out.", "\n Thinking My mind started wandering and thinking about other things during the argument.", "\n\nAll items were presented as statements on a 1--7 Likert scale where 1 meant least agreement and 7 most agreement with the corresponding statement.", "\n\nThe operators that were used for the symbolic regression were: Constant, +, −, ×, /, sqrt, exp, log. ", "The program was run for both *nPhys* and *nVerbal*. ", "The population size (number of formulae per generation) was chosen by Eureqa as 2560. ", "For each analysis the program was run on a 40 core cluster (see Methods) and left running for many hours until the solution set of equations stabilized. ", "The fitness metric used was mean absolute error.", "\n\nWe consider first *nPhys*. ", "The Eureqa program was left to run for more than 2000 core hours. ", "It reported 28 equations. ", "Each has an associated size parameter that represents the complexity of the equation (ranging from 1, least, to 53, most complex), a fitness value, the square of the correlation coefficient between the response variable and the fitted values from the equation, and the Akaike Information Criterion (AIC). ", "The AIC is an information theoretic measure of the relative goodness of fit of a model to the data. ", "Smaller AIC values represent better goodness of fit, taking also into account the complexity of the model. ", "The AIC is often used in model selection procedures, as discussed extensively in [@pone.0052766-Burnham1].", "\n\nThe model with the smallest AICs is shown in Eq (1). ", "Here *group* is 0 for out-group and 1 for in-group. ", "Similarly *LookAt* is 0 for 'off', and 1 for 'on'. ", "The other variables are from the questionnaire ([Table 2](#pone-0052766-t002){ref-type=\"table\"}).", "\n\n(R^2^ = 0.85, AIC = 108, Size = 26).", "\n\n[Figure 3a](#pone-0052766-g003){ref-type=\"fig\"} shows the relationship between the observed and fitted number of interventions based on Eq (1) (the diagram is very similar for all the top fitting equations generated). ", "The high fitting equations all, of course, give similar results and Eq (1) is marginally preferred since it has high explanatory power (in terms of correlation) and the smallest AIC, and on the range of complexity of the models produced is about half way along the scale amongst all generated equations.", "\n\n{#pone-0052766-g003}\n\nThe equation shows a clear distinction between in-group and out-group. ", "For the out-group (group = 0) the entire first term, on the left-hand side of the plus sign, vanishes (20 of the 28 equations generated have this exponential term). ", "For the in-group (group = 1), it can be seen that *LookAt* has a very small but positive influence on the number of interventions but *VictimLooked* has a greater influence. ", "As it ranges from 1 to 7 the number of interventions increases by 0.015\\*exp(*VictimLooked*), which is, for example, 2 for *VictimLooked* = 5, and 16 for *VictimLooked* = 7, other things being equal.", "\n\nThe second term only includes a few of the questionnaire variables. ", "Examining this term, the number of interventions is proportional to concern about the safety of others, and the feeling that the fight should be stopped. ", "It is inversely proportional to the feeling of wanting to get out, and the fear that other people might turn up to make things worse.", "\n\nNow we turn to the number of verbal interventions *nVerbal*, and follow the same analysis. ", "Here the genetic program ran for 1930 core hours. ", "28 equations were produced with size complexity ranging from 1 to 71. ", "The equation with the lowest AIC is shown in Eq (2).", "\n\n(R^2^ = 0.93, AIC = 83, Size = 29).", "\n\nAs before all the high fitting equations give very similar results and we take Eq. (", "2) as representative. [", "Figure 3b](#pone-0052766-g003){ref-type=\"fig\"} shows the plot of fitted by observed values over the data set for Eq. (", "2). ", "Examining the equation we see this time there is no effect of *group*. ", "The number of verbal interventions is proportional to the feeling of the need to stop the fight, and inversely proportional to the fear that other people might arrive and make things worse. ", "Also there is a positive association with participant fears for their own safety. ", "The most interesting variable again is *VictimLooked*, the *belief* that the V avatar was looking towards the participant for help. ", "The variable *MoveAway* is strongly related with *VictimLooked* which must be taken into account otherwise the equations explode into huge values as *VictimLooked* increases. [", "Figure 4](#pone-0052766-g004){ref-type=\"fig\"} shows that there is a very strong positive correlation between these two variables (apart from 1 outlier) (r = 0.71, P = 3.3×10^−7^), with regression line *MoveAway* = −0.38+0.82*VictimLooked*. ", "Moreover 22 out of the 28 equations include the exponential term involving these two variables. ", "We maintain this relationship when examining the effect of *VictimLooked* on *nVerbal* rather than fixing *MoveAway* at a constant value, and taking this into account high values of *VictimLooked* are associated with a larger number of interventions.", "\n\n{#pone-0052766-g004}\n\nThe Interviews {#s2c}\n--------------\n\nAfter the experimental trial there was a short interview with the participants, followed by their debriefing where the purposes of the experiment were explained. ", "The interviews concentrated on several main questions: their feelings and responses during their experience, the extent to which they judged their responses to be realistic, factors that might have increased their intervention, and factors that drew them out of the experience. ", "Summaries of the interviews were coded into key codes and frequency tables constructed, using the HyperResearch software [@pone.0052766-HesseBiber1].", "\n\nWe consider first the responses and feelings of participants during their experience. [", "Table 3](#pone-0052766-t003){ref-type=\"table\"} shows the codes and two example sentences of each code and [Table 4](#pone-0052766-t004){ref-type=\"table\"} the code frequencies.", "\n\n10.1371/journal.pone.0052766.t003\n\n###### Codes for the Interview Questions: What feelings/responses did you have while this was happening?", "\n\n![](", "pone.0052766.t003){#pone-0052766-t003-3}\n\n Code Example Statements\n ----------------------------------- -----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------\n wanted to stop it 1\\. I felt like I would like to stop it (the confrontation) myself, basically back up the person that I was speaking to Arsenal about, protect him.", "\n 2\\. I wanted to calm him down. ", "I wanted to separate them.", "\n uncomfortable 1\\. I felt very uncomfortable.", "\n 2\\. I felt a little bit uncomfortable.", "\n torn about intervening 1\\. I thought about intervening, do something about it, try to calm him down, but probably would have made it worse.", "\n 2\\. I wanted to do something, but I felt I probably couldn't and if I did, I might make things worse to myself. ", "So I just tried to calm him down a little bit, but obviously he didn't want.", "\n would avoid confrontation 1\\. I would avoid confrontation.", "\n 2\\. I would probably have walked out of the CAVE, like I would have done in real life if there was a problem. ", "I was a bit afraid of talking to the man with the white shirt, in case that he would interact with me and get aggressive.", "\n even though VR 1\\. I knew it was VR, and I'm quite surprise how so angry made me feel, the other guy (P), ... I got to the point that I wanted touch him physically or pushing away I felt a bit frustrated I couldn't.", "\n 2\\. I was aware it was a simulation, and I was safe in that respect. ", "I knew it was an aggressive confrontation and I think that has some impact and kind of made me a bit nervous.", "\n anger 1\\. I was quite angry as well, about the way he (P) was treating him, the Arsenal fan.", "\n 2\\. I'm quite surprise how so angry made me feel.", "\n frustration 1\\. I got to the point that I wanted touch him physically or pushing away I felt a bit frustrated I couldn't.", "\n 2\\. ...but it was a kind of frustrating I couldn't because I tried to speak to the guy (P) and he just ignored me.", "\n anxiety or fear 1\\. Very similar feelings as in real life: flustered, panic, helpless and wanting to resolve the situation and not knowing how.", "\n 2\\. Frightened, I was feeling more alert, more mentally prepared for a fight.", "\n helplessness 1\\. Helplessness, unable to help the Arsenal supporter.", "\n 2\\. Helpless because even if was to get involved, I don't know how useful I would be.", "\n confrontational 1\\. I wanted to say I'm wearing an Arsenal shirt as well \\[*he was*\\], so your problem is with me as well, I was just criticizing his argument basically.", "\n 2\\. I thought about punching the aggressor.", "\n uninvolved 1\\. I felt like an observer all the time.", "\n 2\\. To be honest, with VR, I was quite divorced, I was just a kind of watching.", "\n silly or humorous 1\\. I thought it was a bit silly.", "\n 2\\. Humorous.", "\n concerned for or felt sorry for V 1\\. I was concerned for the safety for the man with the red shirt.", "\n 2\\. I felt a bit sorry for the victim, a little compassion for him.", "\n wanted to leave 1\\. I did feel that I wanted to leave.", "\n 2\\. I wanted to leave I didn't want to get involved.", "\n\n10.1371/journal.pone.0052766.t004\n\n###### Frequencies of the Codes in [Table 3](#pone-0052766-t003){ref-type=\"table\"}.", "\n\n![](", "pone.0052766.t004){#pone-0052766-t004-4}\n\n Frequency of statement \n ----------------------------------- ------------------------ --------\n wanted to stop it 16 18\n uncomfortable 2 9\n torn about intervening 11 13\n would avoid confrontation 5 2\n even though VR 0 7\n anger 0 7\n frustration 0 7\n anxiety or fear 14 18\n helplessness 7 7\n confrontational 7 13\n uninvolved 11 0\n silly or humorous 5 0\n concerned for or felt sorry for V 16 0\n wanted to leave 7 0\n **TOTAL no. ", "of statements** **44** **45**\n\nThe impression from the interviews as shown in [Table 4](#pone-0052766-t004){ref-type=\"table\"} is that those in the out-group tended to sympathize with or feel sorry for V. Also many of them wanted to just leave the situation, felt uninvolved, or a few found the situation silly. ", "For those in the in-group it seems to be more anger and frustration that could be the driving force of their intervention, and their response was more likely to be a confrontational one. ", "None of them felt uninvolved, found the situation funny or silly, felt sorry for V or wanted to leave. ", "Some of the in-group expressed surprise at their own responses even though they were aware that it was virtual reality, whereas none of the out-group expressed such surprise. ", "This fits with the fact that many of the out-group felt uninvolved and none of the in-group felt so.", "\n\n[Tables 5](#pone-0052766-t005){ref-type=\"table\"} and [6](#pone-0052766-t006){ref-type=\"table\"} give the results for the interview question regarding the authenticity of response in comparison with reality. ", "We do not show the separate tables for in-group and out-group since there is no difference between them in this regard, although there is some suggestion of a difference between the *LookAt* groups. ", "It seems that those in the *LookAt* 'off' group were more likely to remark on the lack of interaction, and to contrast their behaviour in virtual reality and reality. ", "They were less likely to report their responses as being realistic. ", "In the combined sample just over half found that their responses were realistic.", "\n\n10.1371/journal.pone.0052766.t005\n\n###### Codes for the Interview Question: Were your responses realistic?", "\n\n![](", "pone.0052766.t005){#pone-0052766-t005-5}\n\n Code example statements\n ------------------------------ -----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------\n realistic or quite realistic 1\\. I think that's what I would do in real life.", "\n 2\\. Pretty authentic. ", "I've been in situation like this before, and run your mind afterwards think 'I could have done this, I could have done that, or I should have done this'. ", "but at that time you feel like a deer in the headlights, you are sort of frozen. ", "You want to help, but you don't want that guy to throw a punch on you, it's a fine line.", "\n lacked interaction 1\\. The fact that he (P) didn't recognized me when he came over, I felt I was just watching.", "\n 2\\. I behaved as in real life up to the point that I realized that there was no reaction from them.", "\n contrasts VR and reality 1\\. In real life, I would try to put some distance between them and me, pub fights might be tricky, they might have weapons.", "\n 2\\. I thought about it, but I wasn't sure if they would respond to me. ", "Anyway, in real life I would probably have not intervened. ", "I would have been more scared in real life.", "\n detached 1\\. I was completely detached.", "\n 2\\. It was not authentic at all.", "\n\n10.1371/journal.pone.0052766.t006\n\n###### Frequencies of the Codes in [Table 5](#pone-0052766-t005){ref-type=\"table\"}.", "\n\n![](", "pone.0052766.t006){#pone-0052766-t006-6}\n\n Frequency of statement \n ------------------------------ ------------------------ -------- --------\n realistic or quite realistic 44 62 52\n lacked interaction 15 5 10\n contrasts VR and reality 37 24 31\n detached 4 10 6\n **TOTAL no. ", "of statements** **27** **21** **48**\n\nParticipants were asked what might have increased or decreased their degree of intervention. ", "The results are shown in [Tables 7](#pone-0052766-t007){ref-type=\"table\"} and [8](#pone-0052766-t008){ref-type=\"table\"}. ", "Most frequently they said that if the setup had been more interactive (i.e., the characters responding to their actions after the argument had started) then they would have been more likely to intervene. ", "There were two other aspects that are opposed. ", "On the one side a number of participants said that they would have been more likely to intervene if the perpetrator had become more aggressive. ", "On the other side some participants said that they might have intervened had the perpetrator been less aggressive. ", "Others emphasized that had the victim explicitly called for help they would have been more likely to have intervened. ", "Another important contributory factor could have been greater rapport - for example, the victim having been a friend - or someone in need such as a child.", "\n\n10.1371/journal.pone.0052766.t007\n\n###### Codes for the Interview Question: What would have made it more likely for you to intervene?", "\n\n![](", "pone.0052766.t007){#pone-0052766-t007-7}\n\n Code example statements\n ------------------------ -----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------\n *Aspects that would have increased intervention ...*\n call for help 1\\. If the guy who was threatened would have directly spoken to me.", "\n 2\\. If V would have looked at me and said something to me at some point, something like this \"Can you believe this guy?\"", "\n more interactivity 1\\. If P would have said anything to me.", "\n 2\\. If there had been a reaction from them to my first interventions.", "\n more aggression 1\\. If the aggressor started punching, if the situation would become more physical.", "\n 2\\. If it had turned physical, I would have stepped in. ", "If there was another person joining, I would have definitely stepped in.", "\n more rapport 1\\. If it was a child against a man or a woman against a man, or even if he is a stranger if I maybe spent a match or discuss the football before hand, so there a was a bit of relationship.", "\n 2\\. If the victim was my friend, probably if there was a connection between him and I.\n more realism 1\\. A greater degree of realism.", "\n safety of intervention 1\\. Maybe if the person with the white shirt would have been less aggressive.", "\n 2\\. If P would not have said that he hated Gooners, or if there were more Arsenal fans around.", "\n *Aspect that would have decreased intervention ...*\n knew it was VR 1\\. I knew I was in virtual reality, I wouldn't intervene because I didn't know if I had to.", "\n 2\\. Deep down I knew it was virtual reality.", "\n\n10.1371/journal.pone.0052766.t008\n\n###### Frequencies of the Codes in [Table 7](#pone-0052766-t007){ref-type=\"table\"}.", "\n\n![](", "pone.0052766.t008){#pone-0052766-t008-8}\n\n Code Frequency of statement %\n ----------------------------- --------------------------\n call for help 11\n more interactivity 41\n more aggression 16\n more rapport 11\n more realism 3\n safety of intervention 11\n knew it was VR 8\n **Total no. ", "of statements** **37**\n\nFinally participants were asked to talk about technical factors that drew them out of the experience. ", "It will be seen from the video (Video S1) that, for example, there is no lip sync when the characters talk. ", "This is very obvious when looking at the video, but barely noticeable when immersed in the environment with the life-sized characters. ", "The combination of gesture and natural turn taking in conversation, amongst other things, are probably factors in making this glaring defect not noticeable. ", "Only 5 out of 40 people mentioned the lack of lip sync and it was the fifth most mentioned aspect in this question. [", "Table 9](#pone-0052766-t009){ref-type=\"table\"} shows the list of topics raised by the participants and the number of times they were mentioned. ", "By far the greatest number of issues were concerned with 'plausibility' of the situation itself, and the technical factors tend to come down lower in the list.", "\n\n10.1371/journal.pone.0052766.t009\n\n###### Frequencies of Statements in Response to the Interview Question: What factors tended to draw you out of the experience?", "\n\n![](", "pone.0052766.t009){#pone-0052766-t009-9}\n\n Topic No. ", "of people\n -------------------------------------------------- ---------------\n No other people around 9\n The pub does not look like a real English pub 7\n Dialogue with the victim not realistic 7\n No response from characters during the argument 6\n No background noise or music 5\n No mouth movement of the characters 5\n Lack of sense of touch 5\n Animations not smooth 5\n CAVE walls and edges visible 4\n Aggressor appears from nowhere 3\n Mirror on top not appropriate 2\n Illumination not realistic 2\n Victim appears from nowhere at the start 2\n Anatomical proportions of the characters 2\n No bar staff 2\n Clipping (part of a character going out of view) 1\n Lack of sense of smell 1\n The victim was too defensive 1\n Victim looks ghostly due to Cave rendering 1\n Lack of facial animation 1\n **TOTAL No. ", "of Statements** **71**\n\nDiscussion {#s3}\n==========\n\nThe principal finding of this research with respect to the bystander issue is that participants in the in-group condition made more attempts at physical and verbal intervention than those in the out-group condition. ", "Second, for those in the in-group the number of physical interventions was associated with the belief that the victim was looking towards them for help.", "\n\nThis second finding relies on the important distinction between the experimentally manipulated *LookAt* factor, and the questionnaire report after the experiment about how much the subjects thought that the victim was looking towards them for help (*VictimLooked*). ", "To be clear, *LookAt* refers to whether or not *in fact* the program was making the victim sometimes look towards the participant. ", "The second refers to the *reported belief* of the participant that the victim was looking towards him *for help*. ", "The analysis of covariance (and [Figure 2](#pone-0052766-g002){ref-type=\"fig\"}) showed that the belief that the victim was looking towards the participant for help had a differential effect depending on group. ", "For those in the in-group condition, if they believed that the victim was looking towards them for help their number of interventions tended to be greater. ", "For those in the out-group condition this relationship did not occur. ", "This would not be surprising if it occurred in reality. ", "If you consider you have group affiliation with someone and that person is looking to you for help surely this would be a more important event, more likely to move you to action, than if someone with whom you have no affiliation looks towards you for help. ", "It is especially striking then that this also occurs also in virtual reality (where the only real people were the participants themselves): the more that the participants believed that the victim was looking towards them for help the more often did they intervene - but only those in the in-group condition.", "\n\nThird, the use of symbolic regression as a data exploration method complemented and supported the results found from the classical analysis. ", "Specifically, it provided a further demonstration that those in the in-group and out-group conditions responded quite differently to the influence of the *LookAt* factor and the *VictimLooked* response. ", "Additionally, for those in both in-group and out-group the feeling that they should stop the argument was positively associated with an increased number of physical interventions, as was concern for the safety of the victim. ", "However, the fear that other people might turn up to make the situation worse was inversely related to the number of physical interventions as was the feeling of wanting to get out.", "\n\nThe picture looks different for the number of verbal interventions. ", "Here the group did not seem to play much role. ", "Important factors contributing positively to the number of such interventions were the feelings by participants that they 'should stop it', concern for their own safety, and a strong perception that the victim was looking towards them for help. ", "The factors that contributed negatively were the feeling of wanting to move away from the protagonists, and also the fear that other people might turn up to make the situation worse. ", "However, in the vast majority of equations generated by the symbolic regression the belief that the victim was looking towards them for help is always together with the feeling of wanting to move away from the protagonists. ", "These two variables have opposite effects, but in these data they are very strongly positively correlated. ", "When *VictimLooked* is high, and *MoveAway* is held at its correlated value according to the regression relationship between them, then the number of verbal interventions becomes very high.", "\n\nThe out-group and in-group participants had about the same reported desire to stop the argument, the same level of feeling of being torn about intervening or not, and the same level of anxiety or fear. ", "However, those in the in-group condition expressed greater anger and frustration, whereas those in the out-group condition were more likely to feel sorry for the victim, feel uninvolved or find the situation silly. ", "Those in the in-group condition were more likely to react in a confrontational way compared with those in the out-group, who were looking more to defuse the situation. ", "When we classify the verbal interventions as to whether they were more aimed at defusing the situation or more confrontational, amongst the out-group 17% were confrontational compared to 40% for the in-group, and 73% were defusing utterances compared to 60% for the in-group. ", "These data suggest that the in-group were more likely to respond to the situation through anger and confrontation compared to the out-group, who were either less likely to become involved at all, or more likely to make verbal interventions to defuse the situation. ", "This is not too surprising since by insulting the Arsenal affiliation of the victim in the in-group situation, the perpetrator was also of course indirectly insulting the participants who were all Arsenal supporters.", "\n\nThese data also suggest that physical interventions were more related to the safety of the victim, whereas verbal interventions were more related to safety of the self. ", "The equations for the verbal interventions are more likely to include the 'own safety' than those for the physical interventions.", "\n\nA final point regarding the 'out-group' is that in a sense it is not really an 'out-group' condition. ", "Rather it is simply not 'in-group'. ", "Recalling the fact that all the participants were Arsenal supporters, for the 'out-group' the victim was portrayed as a football supporter of unknown affiliation (though highly unlikely to be Arsenal). ", "The fact that there are clearly different results between the in-group and out-group condition is therefore a quite strong one: it is 'in-group' versus simply not 'in-group'.", "\n\nAn important issue is the extent to which these findings are generalizable. ", "We have shown an example where the group affiliation was a real one: strong supporters of a particular football team. ", "This is unlike many laboratory based experiments where an abstract group affiliation is created for the purposes of the experiment. ", "Our experimental manipulation involved activating the Arsenal affiliation through the virtual character V wearing an Arsenal football shirt, and talking enthusiastically about the club (in-group). ", "The affiliation was not activated for those in the out-group condition, since V was not wearing an Arsenal shirt, and did not engage in enthusiastic conversation about the club. ", "Our interest focused on the extent to which this activated (or not) psychological group affiliation impacted intervention behaviour. ", "Our procedure was therefore designed to generate meaningful psychological group membership - the Arsenal fans were representative of a particular group. ", "Our claim is that it is the perception that the victim belongs to the same group as the participant (in this context he was 'one of us') that leads people to be more likely to intervene. ", "Hence our general hypothesis is that had the group identification been through some other means (social class, race, members of a tennis club, or even arbitrary groups conjured for an experiment) the results would have been similar.", "\n\nIt could be argued that the group of participants might have been too diverse in order to draw these types of conclusions. ", "However, we argue that diversity of the sample is not relevant to this study. ", "Arsenal fans are clearly made up of men, women, Londoners, working class, middle class, and people of different ethnic origins. ", "However, the point is that under some circumstances they come to define themselves as members of the same group (in this case Arsenal fans) - and when this aspect of identity is important to them they are more likely to intervene to help a victim of violence when they think that person shares group membership with themselves in this context. ", "Such group membership can be so powerful that it has been shown to at least temporarily cut across even racial bias in a context where group affiliation was created in a laboratory setting [@pone.0052766-Kurzban1], [@pone.0052766-VanBavel1]. ", "It has further been argued with respect to the famous Milgram obedience and Zimbardo Stanford prison experiments [@pone.0052766-Zimbardo1] that group identification is an excellent predictor of conformity [@pone.0052766-Reicher1], [@pone.0052766-Haslam1]. ", "For example, it was demonstrated, on the basis of the complete set of Milgram's experiments, that the more that subjects identified with the experimenter and his causes (science, answering an important scientific problem) the more likely that they would administer the shocks. ", "On the other hand they would be more likely to disobey the more that they identified with the Learner (representing the general community). ", "Milgram's original set of experiments provided a range of circumstances that led to varying degrees of identification with one of these groups (science or the community), and the degree of obedience varied accordingly.", "\n\nNow we consider how our experiment could be improved. ", "In [@pone.0052766-Slater1] the concept of 'plausibility' of experiences in IVEs was introduced, referring to the illusion of participants that the virtual events are really happening (even though they know that this is not the case). ", "It was argued that plausibility depends at least on three factors: (i) the extent to which there are events that refer personally to the participant, (ii) the extent to which the environment responds to actions of the participant, (iii) and the credibility of the scenario in terms of how much they fit expectations from a similar situation in reality. ", "With respect to the technical setup there were no differences between in-group and out-group, and this is reflected in the fact that there are no differences in reported responses and feelings elicited through the interviews. ", "However, the evidence does suggest ([Table 6](#pone-0052766-t006){ref-type=\"table\"}) a greater tendency for the group with *LookAt* 'on' to say that their responses were realistic, and for those with *LookAt* 'off' to mention the lack of interaction. ", "This is consistent with (i) above.", "\n\nHowever, an overwhelming conclusion from these data is that the plausibility of the experience would be greatly improved through more interactivity (i.e., (ii) above). ", "Recall that there was an interactive episode at the start of the experiment, where in order to establish the in-group and out-group conditions, the eventual victim did have a conversation with the participant. ", "However, once the argument started there was no further interaction in the sense that the virtual characters did not respond to anything that the participant said or did, except for the pre-programmed *LookAt* factor. ", "Another aspect of plausibility that would need to be improved based on the results of this experiment is the credibility of the scenario itself (iii). ", "As seen from [Table 9](#pone-0052766-t009){ref-type=\"table\"} the types of factors that drew people out of the scenario were to do with the setting rather than the technical aspects of the display: no other people around in the pub, it did not look like a real English pub, and the dialogue with the victim itself not being realistic. ", "More than 50% of the statements made in [Table 9](#pone-0052766-t009){ref-type=\"table\"} refer to these types of general credibility, and the remainder are specific technical issues such as 'Illumination not realistic' or 'Lack of facial animation', none of which were commonly stated. ", "By technical issues we refer to aspects of the scenario that require only programming to solve (such as the provision of lip sync). ", "By more general credibility issues we refer to the simulation itself - aspects that require a better understanding of what needs to be there for this to be believable as a fight in a London pub.", "\n\nApart from the introduction of interactivity and other issues relating to credibility, there are several improvements for later versions of this experiment. ", "For example, we have not said anything about the role of the social identity of the perpetrator with respect to the participant. ", "Moreover there are clearly other issues involved - such as participant fear of being harmed by the perpetrator. ", "This has not been considered at all, but could also be incorporated into an experiment through manipulation of the appearance of the perpetrator (for example, to look more or less menacing). ", "Finally, future experiments will also manipulate the number of bystanders, and thus directly tackle the question of the role of the number of bystanders in intervention.", "\n\nIn this paper we have shown that immersive virtual reality can be usefully exploited to study the likelihood of bystander intervention in interpersonal violent incidents. ", "The paradigm allows the investigation of what participants did do and think during an actual experience involving violence rather than their opinion of what they might do or what they think others might do - whether based on watching a video or on a verbal description of a situation [@pone.0052766-Banyard1]. ", "Moreover we have exploited the powerful tool of genetic programming to explore these data in a deeper way than is possible with normal statistical methods, highlighted by the elegant distinction between the in-group and out-group conditions shown in Eq. (", "1).", "\n\nOf course, there is still no proof that what participants would do in a physically real situation would match that which we find in virtual reality. ", "However, as reported in the introduction to this paper there is evidence to suggest that people do respond realistically in IVEs. ", "In fact since these experiments can never be carried out in reality, ultimately the question of the validity of people's responses to the virtual situation can never be known through laboratory based experiments of any kind. ", "However, our approach can be used in the process of constructing theories, that can then be further tested with the use of experiments in virtual reality, and moreover ultimately examine how well these theories fit what might be found in actual experiences in the field.", "\n\nTo conclude, we note that the findings for this type of research can also have implications for policy. ", "For example, by creating an atmosphere where it is thought that not running away from a violent scene is the right thing to do, and by encouraging people to ask for help when they are victims of such a situation, it may be possible to engineer pro-social behaviour in specific circumstances where this is thought desirable by policy makers, and actually to manipulate the same variables to avoid it in other situations (e.g., \"do not approach this man since he is considered armed and dangerous\"). ", "Here it would be a question of using the group to enforce social norms for the prevention of violent behaviour. ", "The key to tackling the so called 'walk--on-by' society lies in using the power of group identification to promote social solidarity -- and to persuade and empower bystanders to intervene, in situations where this is considered by the authorities to be appropriate.", "\n\nMethods {#s4}\n=======\n\nEthics Statement {#s4a}\n----------------\n\nThe experiment was approved by the UCL Research Ethics Committee, and was carried out under written informed consent from each participant.", "\n\nThe Virtual Reality System {#s4b}\n--------------------------\n\nA four screen projection system driven by a 5 PC cluster was used. ", "We refer to this by generic name 'Cave' being the type of system described in [@pone.0052766-CruzNeira1]. ", "The Cave has three 3 m×2.2 m back-projected screens: front, left, and right, and a 3 m×3 m front projection surface on the floor. ", "The computers in the cluster contain Intel Pentium 3.2 GHz processors with 1 gigabyte of RAM and Nvidia Quadro FX 5600 graphics cards. ", "The display resolution is 1024×768 pixels for each screen.", "\n\nThe participants were fitted with Crystal Eyes shutter glasses that were synchronized with the projectors, delivering active stereo at 45 Hz each eye. ", "Head-tracking was performed with an InterSense IS-900 tracking device.", "\n\nThe program was written using the XVR programming platform as described in [@pone.0052766-Tecchia1]. ", "The virtual characters were animated using the Hardware Accelerated Library for Character Animation, HALCA [@pone.0052766-Gillies1].", "\n\nThe Scenario {#s4c}\n------------\n\nTwo professional actors were hired to act the scene for the character animation motion capture. ", "A Vicon motion capture system with 6 infrared cameras was used to capture their motions simultaneously. ", "Sound was also recorded at the same time using Audacity software (audacity.sourceforge.net) with two wireless microphones attached to each actor. ", "This raw data was then cleaned up, synchronized and split into pieces so that each one could be later assigned to a button on the interface to be played when needed during the study.", "\n\nDuring the experiment the free-flowing conversation between the participant and V was achieved by operator control. ", "A number of utterances had been recorded for V, each one making a statement or asking a question of the participant. ", "Each such utterance was selected interactively by a hidden operator who could hear the responses of the participant. ", "The operator sat by a computer screen, and all the phrases were represented visually as selectable buttons on the screen. ", "When a button was selected (by point-and-click with the mouse) then V would say the phrase with a corresponding animation.", "\n\nThere was a defined script that the operator followed, but when the participant said something that fell outside of the script, then a number of general phrases could be selected by the operator in order to keep the conversation going in a natural way. ", "For example, if the participant said something out of line, the operator could select a phrase such as \"Totally agree with you\" which would then be said by V. The overall effect for most of the time for most participants sounded as if it were a normal conversation between two people.", "\n\nProcedures and Scenario Details {#s4d}\n-------------------------------\n\n40 male participants were recruited by advertisements around the UCL campus, where we specified that we needed football supporters ('soccer' in American usage). ", "They were required to complete a questionnaire that asked about their favourite team in the English Premier League and how much they supported this team. ", "We only recruited those who supported Arsenal Football Club to the level of at least 4 on a scale from 1 (not at all) to 7 (very much so). ", "They were paid £7 (\\$10--12) for their participation. ", "The experiment was approved by the UCL Research Ethics Committee.", "\n\nUpon arrival at the laboratory participants were given a short questionnaire to complete that obtained information as to their English proficiency, medication, recent alcohol intake, degree of computer game playing, and past familiarity with virtual reality. ", "Their age was obtained at the recruitment stage, in order to ensure that no one under 18 would be recruited.", "\n\nAfter this they were given an information sheet to read, and the same information was again told to them verbally. ", "This described the equipment that would be used. ", "It also warned them that some people experience a degree of nausea in virtual reality systems, and that they were free to withdraw at any time without giving reasons. ", "They were told that they would virtually visit a bar where something was to take place and that they should feel free to interact with other people there. ", "They were warned that experience was going to involve discussion about football and the language and situation depicted was realistic. ", "It included the statement \"If you are someone who would be put off by witnessing realistic scenes that might include bad language or aggressive behaviour, then you should not take part in this experience.\"", "\n\nAfter participants had agreed to take part they were given a consent form to read and sign, and again told that they were free to leave the experiment without having to give reasons. ", "They were then invited to take off their shoes to enter the Cave, and put on the eyeglasses.", "\n\nThe participants entered the virtual reality and were asked to look around and observe the scene, which was a bar of size 4.5 meters deep by 18 meters wide. ", "The participant was then left alone in the bar having been instructed to look around for items related to football for 2 minutes. ", "This allowed them to become familiar with the bar and accommodate to the virtual reality display including the shutter glasses and the overall brightness of the scene. ", "After this time, a virtual character entered the scene and started a conversation with the participant by saying \"You alright mate?\" ", "in the in-group version, and \"Hi, how is it going?\" ", "in the out-group one.", "\n\nNot every conversation was the same across all participants in each detail due to different responses by the participants. [", "Table S1](#pone.0052766.s002){ref-type=\"supplementary-material\"} shows two such conversations, one when V is an Arsenal football club fan (in-group) and the other when just a general football fan (out-group).", "\n\nAfter about 2 minutes of this conversation they were interrupted by another character (P) that had been sitting by the bar, who stood up and approached V and said to him \"Oy! ", "Have you got a problem?\" ", "and then accused V of \"staring\" at him ([Figure 1](#pone-0052766-g001){ref-type=\"fig\"}). ", "This quickly became a strong verbal attack on V, with V remaining submissive throughout. ", "Eventually after 140 s the P avatar started to violently push V, at which point the program ended and the participant took off the glasses and left the Cave.", "\n\nThe participants then were asked to complete a questionnaire about their various responses to the situation ([Table 2](#pone-0052766-t002){ref-type=\"table\"}) followed by an interview and debriefing where the purposes of the experiment were explained to them, and they were asked not to discuss it with others for 3 months in case they spoke with a future participant. ", "They were then paid the £7 (\\$10) and the experimental trial was complete.", "\n\nExperimental Design {#s4e}\n-------------------\n\nThe experimental design was 2×2 between groups, with 10 participants arbitrarily assigned to each of the 4 cells. ", "The two factors were *Group* (in-group/out-group) and *LookAt* (off/on). ", "In-group was signified by V wearing an Arsenal football shirt, and maintaining an initial enthusiastic conversation about Arsenal. ", "Out-group was signified V wearing a football shirt the same as for the in-group except without the Arsenal insignia, and during the conversation his responses were neutral and did not show much interest in the team ([Table S1](#pone.0052766.s002){ref-type=\"supplementary-material\"}).", "\n\n*LookAt* referred to whether V had been programmed to occasionally look at the participant during the confrontation or not. ", "If 'yes' then 5 times during the confrontation V looked toward the participant for 3 seconds. ", "This was possible since the head-tracking streamed continual real-time data to the computer program about the position and orientation of the participant's head. ", "If 'no' there was no particular programmed action that would lead V to look towards the participant, but this may occasionally have occurred by chance (depending on where the participant was standing at the time).", "\n\nResponse Variables {#s4f}\n------------------\n\nOur major response variable of interest concerned the extent to which participants intervened during the confrontation. ", "A video camera was mounted above the Cave looking down at the scenario and recorded each entire experimental trial. ", "The video for two participants could not be analyzed, one for a participant in condition 'out-group' and *LookAt* 'off' and the other in condition 'out-group' and *LookAt* 'on'. ", "These participants were eliminated from all analysis involving counts of the number of interventions. ", "The videos were analyzed independently by two different people, covering the time from when P first accosted V to the end. ", "They had been instructed to count the number of verbal (*Verbal*) and physical (*Physical*) interventions.", "\n\nVideo Analysis {#s4g}\n--------------\n\n[Figure S1](#pone.0052766.s001){ref-type=\"supplementary-material\"} shows two stills from one of the recordings - with first P at the start of the confrontation and then a moment while the participant was intervening by placing himself between V and P and raising his hand.", "\n\nFirst one of the experimenters carried out a review of all the videos noting the number of times that the participant said something to the virtual characters (variable *nVerbalApprox*) and the number of physical interventions - meaning the number of times that the participant moved closer to the characters or reached out towards them (*nPhysApprox*). ", "Second and independently someone not associated with the research, and not knowing its purposes was hired to carry out a complete video analysis using the ELAN system and as paid work ([www.lat-mpi.eu/tools/elan](http://www.lat-mpi.eu/tools/elan)).", "\n\nThe instructions were to record the total number of utterances during the relevant period (*nVerbalElan*) and also the number of physical interventions (*nPhySElan*). ", "The instructions for the physical interventions were to regard as an intervention or an attempt at intervention an action accompanied with verbal intervention or reaching out to either of the avatars. ", "When not accompanied by these it was considered intervention if the participant was walking with purpose towards the avatars or was followed by another form of physical or verbal intervention. ", "Walking or stepping towards the avatars was not considered to be intervention if the participant took a step forwards, backwards, or to the left or right when far from the avatars, if they walked or stepped forwards and this was followed by them standing passively watching the avatars or when they were walking around the environment and the avatars and appeared to be simply investigating the surroundings.", "\n\nAlthough the procedures used for the approximate and ELAN based intervention recordings were not the same the results are strongly correlated. [", "Table S2](#pone.0052766.s003){ref-type=\"supplementary-material\"} shows the correlation coefficients between the various measures of intervention. ", "The approximate and ELAN based methods were consistent, with highly significant positive correlations.", "\n\nSince the ELAN based method was carried out by someone not involved in the research team and more thorough, with the notion of 'intervention' more rigorously and conservatively defined, we base all analysis on this, so that *nPhys* = *nPhysElan*, and *nVerbal* = *nVerbalElan*.", "\n\nSupporting Information {#s5}\n======================\n\n###### \n\n**A still from a video recording from above.** (", "a) The participant can be seen near the centre with the victim to his left, and the perpetrator to his right. (", "b) The participant has stepped between the victim and perpetrator standing in front of the latter and raising his hand.", "\n\n(TIF)\n\n###### \n\nClick here for additional data file.", "\n\n###### \n\n**Examples of Conversations between the Virtual Character V, and participant S.**\n\n(DOCX)\n\n###### \n\nClick here for additional data file.", "\n\n###### \n\n**Pearson Correlation Coefficients Between the Intervention Variables.**", "\n\n(DOCX)\n\n###### \n\nClick here for additional data file.", "\n\n###### \n\n**Degree of Support for the Arsenal Football Club.**", "\n\n(DOCX)\n\n###### \n\nClick here for additional data file.", "\n\n###### \n\n**MANOVA for the Number of Physical and Verbal Interactions.**", "\n\n(DOCX)\n\n###### \n\nClick here for additional data file.", "\n\n###### \n\n**Symbolic Regression.**", "\n\n(DOCX)\n\n###### \n\nClick here for additional data file.", "\n\n###### \n\n**The experimental scenario for the in-group condition.**", "\n\n(MP4)\n\n###### \n\nClick here for additional data file.", "\n\nWe thank Victoria Stafford for the ELAN video analysis and the actor Morgan Roberts who performed in the video.", "\n\n[^1]: **Competing Interests:**The authors have declared that no competing interests exist.", "\n\n[^2]: Conceived and designed the experiments: MS ML. ", "Performed the experiments: AR DS RS. ", "Analyzed the data: MS. ", "Wrote the paper: MS ML AR RS DS JJZ CC. ", "Computer animation: RS JJZ.", "\n" ]

{ "pile_set_name": "PubMed Central" }

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[ "Q:\n\nHow to prevent font-size change of child element to stretch parent container in CSS\n\nI have a simple nav bar:\n<nav>\n <a href=\"#\">Home</a>\n <a href=\"#\">About</a>\n <a href=\"#\">Register</a>\n</nav>\n\nwith following CSS for nav and a tags:\nnav\n{\n background-color:black;\n color:white;\n text-align:center;\n font-size:150%;\n padding:2%; \n}\nnav a\n{\n text-decoration:none;\n color:white;\n padding:2%; \n}\nnav a:hover\n{\n background-color:white;\n color:black;\n font-size:120%; \n}\n\nIt looks pretty much as I want it to, except that nav container stretches on hover, because font size is larger. ", "How can I prevent nav from stretching?", "\n\nA:\n\nThere is an easier and prettier way. ", "You should try this instead. ", "\nnav\n{\n background-color:black;\n color:white;\n text-align:center;\n font-size:150%; \n}\nnav a\n{\n text-decoration:none;\n color:white;\n padding:2%; \n display:inline-block;\n}\nnav a:hover\n{\n background-color:white;\n color:black;\n transform: scale(1.2);\n -webkit-transform: scale(1.2);\n -ms-transform:scale(1.2);\n}\n\n" ]

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