Miracle-dz/UniFAR-FTUs huggingface dataset:
Use of a Two-Color Genetic Screen To Identify a Domain of the Global Regulator Lrp That Is Specifically Required forpap Phase Variation

ABSTRACT The global regulator Lrp plays a central role as both a repressor and an activator in Pap phase variation. Unlike most other members of the Lrp regulon such as ilvIH, activation ofpapBA transcription requires the coregulator PapI and is methylation dependent. We developed a two-color genetic screen to identify Lrp mutations that inhibit Pap phase variation but still activate ilvIH transcription, reasoning that such mutations might identify PapI binding or methylation-responsive domains. Amino acid substitutions in Lrp at position 126, 133, or 134 greatly reduced the rate of Pap switching from phase off to phase on but had much smaller effects on ilvIH transcription. In vitro analyses indicated that the T134A and E133G Lrp variants maintained affinities for pap and ilvIH DNAs similar to those of wild-type Lrp. In addition, both mutant Lrp’s were as responsive to PapI as wild-type Lrp, evidenced by an increase in affinity forpap Lrp binding sites 4, 5, and 6. Thus, in vitro analyses did not reveal the step(s) in Pap phase variation where these Lrp mutants were inhibited. In vivo analyses showed that both the T134A and E133G Lrp mutants activated transcription of a phase-on-lockedpap derivative containing a mutation in Lrp binding site 3. Further studies indicated that the T134A Lrp mutant was blocked in a step in Pap phase variation that does not involve PapI. Our data suggest that these mutant Lrp’s are defective in a previously unidentified interaction required for the switch from the phase-off to the phase-on pap transcription state.
Immunophenotypic and Genotypic Characterization of Progression in Follicular Lymphomas

Progression of follicular lymphomas (FLs) is often accompanied by a spectrum of histologic changes and an aggressive clinical course. Although molecular alterations have been implicated in this event, the underlying factors are largely unknown. We studied the expression of selected tumor suppressor genes (P53 and retinoblastoma [RB]), oncogenes (MYC and BCL2), and a transferrin-receptor related protein (Trump) in sequential biopsies in 16 patients. Eleven patients progressed from grade I or II FL to aggressive B-cell lymphomas with diffuse morphology, whereas 5 patients presented with diffuse aggressive lymphomas and recurred with indolent lymphomas. Immunoreactivity for P53 correlated with higher histologic grade in lymphomas progressing from indolent to aggressive; however, only 1 patient who presented with aggressive lymphoma demonstrated a P53 gene mutation. Neither P53 immunoreactivity nor genotypic alterations correlated with presentation with an aggressive histology and relapse with FL. Growth fraction, as assessed by Ki-67 staining, and Trump expression correlated with histologic grade. Immunoreactivity for RB, BCL2, and MYC was seldom associated with progression. Eight of 9 cases tested exhibited identical immunoglobulin heavy and light chain rearrangements or identical BCL2 gene rearrangements in the sequential lymphomas. We conclude that P53 and Trump protein expression and proliferation activity correlate with histologic grade, but not with recurrence or progression of FL. Our results further indicate that progression of FL to diffuse aggressive lymphomas and presentation of an aggressive B-cell lymphoma followed by FL are clonally related.
Pyrrolidine Dithiocarbamate Reduces the Severity of Cerulein-Induced Murine Acute Pancreatitis

The nuclear factor-&kgr;B (NF-&kgr;B) is a transcription factor that plays a pivotal role in the induction of genes involved in the response to injury and inflammation. Dithiocarbamates are antioxidants that are potent inhibitors of NF-&kgr;B. This study tested the hypothesis that pyrrolidine dithiocarbamate (PDTC) attenuates experimental acute pancreatitis. Intraperitoneal injection of cerulein in mice resulted in severe, acute pancreatitis characterized by edema, neutrophil infiltration, tissue hemorrhage and necrosis, and elevated serum levels of amylase and lipase. Infiltration of pancreatic and lung tissue with neutrophils (measured as increase in myeloperoxidase activity) was associated with enhanced lipid peroxidation (increased tissue levels of malondialdehyde). Immunohistochemical examination demonstrated a marked increase in immunoreactivity for nitrotyrosine and intracellular adhesion molecule-1 in the pancreas and lung of cerulein-treated mice. In contrast, the degree of 1) pancreas and lung injury, 2) upregulation/expression of intracellular adhesion molecule-1, 3) staining for nitrotyrosine, and 4) lipid peroxidation was markedly reduced by pretreatment with PDTC. This study demonstrates that prevention of the activation of NF-&kgr;B by PDTC ameliorates the tissue injury associated with experimental murine acute pancreatitis and provides an important insight into the molecular biology of acute pancreatitis.
Correlations between dynamic contrast‐enhanced magnetic resonance imaging–derived measures of tumor microvasculature and interstitial fluid pressure in patients with cervical cancer

To correlate permeability (rktrans), extracellular volume fraction (rve), relative to muscle and initial area under the enhancement curve (IAUC60m) determined by dynamic contrast‐enhanced magnetic resonance imaging (DCE‐MRI) with in vivo measurements of interstitial fluid pressure (IFP) in patients with cervical cancer.
A hormone-encoding gene identifies a pathway for cardiac but not skeletal muscle gene transcription

In contrast to skeletal muscle, the mechanisms responsible for activation and maintenance of tissue-specific transcription in cardiac muscle remain poorly understood. A family of hormone-encoding genes is expressed in a highly specific manner in cardiac but not skeletal myocytes. This includes the A- and B-type natriuretic peptide (ANP and BNP) genes, which encode peptide hormones with crucial roles in the regulation of blood volume and pressure. Since these genes are markers of cardiac cells, we have used them to probe the mechanisms for cardiac muscle-specific transcription. Cloning and functional analysis of the rat BNP upstream sequences revealed unexpected structural resemblance to erythroid but not to muscle-specific promoters and enhancers, including a requirement for regulatory elements containing GATA motifs. A cDNA clone corresponding to a member of the GATA family of transcription factors was isolated from a cardiomyocyte cDNA library. Transcription of this GATA gene is restricted mostly to the heart and is undetectable in skeletal muscle. Within the heart, GATA transcripts are localized in ANP- and BNP-expressing myocytes, and forced expression of the GATA protein in heterologous cells markedly activates transcription from the natural cardiac muscle-specific ANP and BNP promoters. This GATA-dependent pathway defines the first mechanism for cardiac muscle-specific transcription. Moreover, the present findings reveal striking similarities between the mechanisms controlling gene expression in hematopoietic and cardiac cells and may have important implications for studies of cardiogenesis.
The mutational spectrum of ENPP1 as arising after the analysis of 23 unrelated patients with generalized arterial calcification of infancy (GACI)

Generalized arterial calcification of infancy (GACI), is characterized by calcification of the internal elastic lamina of large and medium‐sized arteries and stenosis due to myointimal proliferation. Although survival to adulthood has been reported, most patients die within the first six months of life. Recently, we found mutations of ENPP1 coding for ecto‐nucleotide pyrophosphatase/phosphodiesterase 1 to be associated with GACI in 8 of 11 families. In this study, we analyzed ENPP1 in affected individuals of another 12 unrelated families. We identified 11 novel homozygous or compound heterozygous mutations in 10 of the 12 new families. The mutations (1 nonsense, 7 missense, 1 single amino acid deletion, and 2 frame shift mutations) were scattered over the whole coding region with a slightly more condensed distribution within the catalytic and nuclease‐like domain as compared to the first survey. In this study, three mutations were found repeatedly in apparently unrelated patients, 7 x c.913C>A (p.Pro305Thr) and c.2680C>T (p.Arg888Trp) as well as c.2320C>T (p.Arg774Cys) each twice. However, haplotype analysis suggested a founder effect of British extraction for mutation c.913C>A (p.Pro305Thr). The fact that the two other mutations c.2680C>T (p.Arg888Trp) and c.2320C>T (p.Arg774Cys) occurred twice within a single allele also suggests a single founder. This study confirms the role of ENPP1 mutations as the main cause of GACI and adds considerably to the mutational spectrum of ENPP1. © 2004 Wiley‐Liss, Inc.
ISOLATION AND CHARACTERIZATION OF A NEW TRYPSIN-LIKE ENZYME FROM: Tenebrio molitor L. LARVAE

A trypsin-like enzyme (TLE) was separated and purified from Tenebrio molitor larval midgut enzyme solution by ion-exchange chromatography on a DEAE-cellulose column. The purified enzyme was found to be a homogeneous protein by electrophoresis in polyacrylamide gels and on cellulose acetate strips, by electrofocusing in polyacrylamide gels and by SDS-polyacrylamide gel electrophoresis. Its molecular weight was estimated to be 18300 by ultracentrifugal analysis and 24300 on SDS-polyacrylamide gel electrophoresis. It has an isoelectric point 8.0, it contains only four half-cystine residues per molecule and the NH2-terminal amino acid is isoleucine. TLE possesses a high degree of specificity towards trypsin synthetic substrates such as N-α-benzoyl-DL-arginine p-nitroanilide (BAPNA), p-tosyl-L-arginine methyl ester (TAME) and poly-L-lysine hydrobromide. The optimal pH for TLE activity was found to be 8.0 and the optimal temperature 50° C. Its Km value when assayed on BAPNA was 0.93mM and on TAME 0.08mM. TLE is stable at neutral pHs and its activity is not affected by Ca2+ and by 0.01 M 1, 4-dithiothreitol (DTT). It is inactivated by DFP and tosyl-L-lysine chloromethylketone (TLCK) and is fully inhibited by the naturally occurring trypsin inhibitors such as trypsin-and α-chymotrypsin inhibitor (AA) from soybeans, basic pancreatic trypsin inhibitor (BPTI), chick peas trypsin and chymotrypsin inhibitor (CI) and crystalline soybean trypsin inhibitor (CSBTI), forming with them complexes in a molar ratio of 1:1. The Ki value for AA with BAPNA as substrate is 5.87 10-7 M and for BPTI 7.92 10-7M. No common antigenic determinants were noted between TLE and bovine trypsin. This finding together with the relatively low number of -S-S- bonds in the TLE molecule indicate that TLE differs in conformation from bovine trypsin.
Effect of DSS‐induced colitis on visceral sensitivity to colorectal distension in mice

Abstract The present study aimed at evaluating the effect of dextran sodium sulphate (DSS)‐induced colitis on visceral sensitivity, measured as the visceromotor response (VMR) to colorectal distension (CRD) in BALB/c and C57Bl/6 male mice. Inflammation was induced by the addition of 4% DSS to the drinking water for 5 (C57Bl/6) or 6–7 days (BALB/c). Parallel groups were used to monitor histopathological changes and visceral sensitivity. Pseudo‐affective visceral pain responses were evoked using an increasing phasic CRD paradigm (10–60 mmHg) in conscious mice on predetermined days (pretreatment controls, 12, 16, 20, 30, 40 and 51). In both mouse strains, significant histopathological changes developed between days 2 and 5 of DSS treatment, and persisted until day 12 (P < 0.05). On day 15, inflammatory scores were reduced by about 50%. Despite evidence of inflammation in DSS‐treated mice, no differences could be shown in the VMR to CRD between DSS‐treated mice and controls at any time point tested. In addition, no differences were seen before and after DSS treatment in the same group of mice. In conclusion, these data suggest that DSS‐induced colonic inflammation does not affect the visceral sensitivity to CRD, neither at short or long term, in BALB/c or C57Bl/6 male mice.
Testosterone:estradiol ratio changes associated with long-term tadalafil administration: a pilot study.

INTRODUCTION It has been reported that lack of sexual activity due to erectile dysfunction (ED) may be associated with testosterone (T) decline. AIM To investigate whether the known changes in sex hormones associated with resumption of sexual activity are sustained in the long term. MAIN OUTCOME MEASURES Primary endpoints were variations from baseline of steroid hormones: total T, free T (f T), and estradiol (E). Secondary endpoints were variations of erectile function domain scores at International Index of Erectile Function-5 (IIEF-5). METHODS In an open-label fashion, 20 patients (mean age 54.8 +/- 8.4 years) received tadalafil 10-20 mg on demand for 12 months. Exclusion criteria were those reported for phosphodiesterase inhibitors, including hypogonadism and hyperprolactinemia. RESULTS Tadalafil assumption was safe and well tolerated (overall adverse effects in 15% of patients) and none discontinued medication. A significant decrease in E levels occurred at the end of the study (from 19.9 +/- 9.6 to 16.6 +/- 8.1 ng/dL, P = 0.042 vs. baseline), with parallel increase in the T:E ratio (26.3 +/- 15.3 to 32.6 +/- 17.7, P = 0.05), whereas no changes in T and f T serum levels were observed, respectively (411.4 +/- 131.4 to 434.2 +/- 177.1 ng/dL and 47.7 +/- 15.3 to 49.9 +/- 19.1 pmol/L, not significant). Interestingly, nonparametric subgroup analysis for related samples revealed that E decrease was detectable only in lean (N = 14) but not in obese (N = 6, body mass index > 27.5 kg/m2) subjects (17.8 +/- 10.1 vs. 13.5 +/- 6.8, P < 0.05). A net increase in IIEF-5 scores was observed at the endpoint (13.7 +/- 5.9 vs. 25.7 +/- 2.9, P < 0.0001). CONCLUSIONS Sustained improvement in sexual function after 12 months of tadalafil administration is associated with increased T:E ratio mainly related to reduction of E levels. We hypothesize that androgen-estrogen cross-talk and possible inhibition of aromatase activity during chronic exposure to tadalafil might have a role in the regulation of erectile function.
Apolipoprotein E immunoreactivity in neurons and neurofibrillary degeneration of aged non‐demented and alzheimer's disease patients

Apolipoprotein E (ApoE) genotype is a risk factor for Alzheimer's disease (AD) but its relationship with neurofibrillary degeneration remains obscure. To further analyze this relationship, hippocampal, entorhinal, temporopolar, and insular cortices of 10 non‐demented and 7 Alzheimer disease brains were studied with both light and electron microscopy. Focus was directed on pretangles and neurons starting to accumulate tangles. ApoE immunolabeling in neurons and tangles was independent of ApoE individual genotype. The majority of the neurons in all of the brains were ApoE‐negative, but virtually every brain also contained groups of ApoE‐immunoreactive neurons, with diffuse cytoplasmic labeling. Most of the ApoE‐positive tangles were extracellular, but a few tangles were shown to be intraneuronal when studied ultrastructurally. No ApoE immunoreactivity was found in neuropil threads, as well as in neurites associated with senile plaques. Double protocols with both AT‐8 and anti‐ApoE antibodies, performed to determine whether ApoE‐positive neurons were pretangle neurons, did not detect cytoplasmic AT‐8 in ApoE‐positive neurons. Though careful electron microscopy studies found ApoE reaction product in an occasional ApoE‐positive pretangle‐like neuron and a few intracellular tangles, these findings do not support that ApoE is necessary for the accumulation of hyperphosphorylated tau protein. The more consistent colocalization of anti‐ApoE and AT‐8 in extracellular tangles reveals that ApoE mainly binds to tangles once they are in the extracellular space, in a manner similar to that described for amyloid fibrils. Microsc. Res. Tech. 55:48–58, 2001. © 2001 Wiley‐Liss, Inc.
Membrane Structure: Some General Principles

The arrangement of lipids and some proteins in the erythrocyte membrane has been discussed. The conclusions from this are listed here as a set of general guidelines for the structure of membranes of higher organisms: some of these rules may be wrong. But at this stage it seems useful to sharpen our thoughts in this way and thereby focus attention on various specific points. 1) The basis of a membrane is a lipid bilayer with (i) choline phospholipids and glycolipids in the external half and (ii) amino (and possibly some choline) phospholipids in the cytoplasmic half. There is effectively no lipid exchange across the bilayer (unless enzymatically catalyzed) (68). 2) Some proteins extend across the bilayer. Where this is so, they will in general have carbohydrate on their surface remote from the cytoplasm. This carbohydrate may prevent the protein diffusing out of the membrane into the cytoplasm; it acts as a lock on the protein. 3) Just as lipids do not flip-flop, proteins do not rotate across the membrane. Lateral motion or rotation of lipids and proteins in the plane of the bilayer may be expected. 4) Most membrane protein is associated with the inner, cytoplasmic, urface of the membrane. Proteins are not usually associated exclusively with the outer half of the lipid bilayer. 5) Membrane proteins are a special class of cytoplasmic proteins, not of secreted proteins.
A randomized clinical trial of high‐intensity warfarin vs. conventional antithrombotic therapy for the prevention of recurrent thrombosis in patients with the antiphospholipid syndrome (WAPS) 1

Summary. Background: The optimal intensity of oral anticoagulation for the prevention of recurrent thrombosis in patients with antiphospholipid antibody syndrome is uncertain. Retrospective studies show that only high‐intensity oral anticoagulation [target international normalized ratio (INR) >3.0] is effective but a recent randomized clinical trial comparing high (INR range 3.0–4.0) vs. moderate (INR 2.0–3.0) intensities of anticoagulation failed to confirm this assumption. Methods: We conducted a randomized trial in which 109 patients with antiphospholipid syndrome (APS) and previous thrombosis were given either high‐intensity warfarin (INR range 3.0–4.5, 54 patients) or standard antithrombotic therapy (warfarin, INR range 2.0–3.0 in 52 patients or aspirin alone, 100 mg day−1 in three patients) to determine whether intensive anticoagulation is superior to standard treatment in preventing symptomatic thromboembolism without increasing the bleeding risk. Results: The 109 patients enrolled in the trial were followed up for a median time of 3.6 years. Mean INR during follow‐up was 3.2 (SD 0.6) in the high‐intensity warfarin group and 2.5 (SD 0.3) (P < 0.0001) in the conventional treatment patients given warfarin. Recurrent thrombosis was observed in six of 54 patients (11.1%) assigned to receive high‐intensity warfarin and in three of 55 patients (5.5%) assigned to receive conventional treatment [hazard ratio for the high intensity group, 1.97; 95% confidence interval (CI) 0.49–7.89]. Major and minor bleeding occurred in 15 patients (two major) (27.8%) assigned to receive high‐intensity warfarin and eight (three major) (14.6%) assigned to receive conventional treatment (hazard ratio 2.18; 95% CI 0.92–5.15). Conclusions: High‐intensity warfarin was not superior to standard treatment in preventing recurrent thrombosis in patients with APS and was associated with an increased rate of minor hemorrhagic complications.
Ethics and Issues of Secondary Prevention Efforts in Child Sexual Abuse

This article discusses the ethical, practical, and moral issues surrounding secondary prevention efforts of child sexual abuse from a professional and practice-based perspective. Transcripts of a semistructured consultation event with n = 15 international experts on the secondary prevention of child sexual abuse were analysed using thematic qualitative analysis. The research identified four main critical areas linked to secondary prevention efforts, including, the psychology of self-reporting and disclosure; the interaction with and within existing legal, social, and professional frameworks; the scale and type of an appropriate response; and potential hurdles (i.e., within media, public, politics). The article outlines these areas, highlighting participant perspectives on risk-enhancing and mitigating factors for each domain.
Information Production in Multiple Lead Underwriter IPOs

We use textual analysis techniques to examine how the size of IPO lead underwriting syndicate influences the information production decisions of underwriters. Our results show that IPO prospectuses produced by multiple lead underwriters (MLUs) contain more informative content and use more readable language. These findings imply that MLUs produce and discloses more pricing information during the pre-filing period. We also find that MLUs produce less information during bookbuilding and provide less support for partial adjustment phenomenon. Together, our results suggest that MLUs can substitute the costly bookbuilding with the pre-filing information services.
A Gap-Based Framework for Chinese Word Segmentation via Very Deep Convolutional Networks

Most previous approaches to Chinese word segmentation can be roughly classified into character-based and word-based methods. The former regards this task as a sequence-labeling problem, while the latter directly segments character sequence into words. However, if we consider segmenting a given sentence, the most intuitive idea is to predict whether to segment for each gap between two consecutive characters, which in comparison makes previous approaches seem too complex. Therefore, in this paper, we propose a gap-based framework to implement this intuitive idea. Moreover, very deep convolutional neural networks, namely, ResNets and DenseNets, are exploited in our experiments. Results show that our approach outperforms the best character-based and word-based methods on 5 benchmarks, without any further post-processing module (e.g. Conditional Random Fields) nor beam search.
The influence of APOAV polymorphisms (T‐1131>C and S19>W) on plasma triglyceride levels and risk of myocardial infarction

The importance of an APOAV gene for plasma triglyceride level determination has been shown on transgenic and knockout mice. We examined whether APOAV variants are associated with plasma triglyceride levels and risk of myocardial infarction (MI). We have evaluated the influence of APOAV polymorphisms (T‐1131>C and S19>W) on plasma triglycerides in 1191 males and 1368 females representatively selected from the Czech population. Triglycerides have been analysed in 1997 and 2001. Subsequently, we have analysed the genotype frequencies of the APOAV polymorphism in 435 patients with MI. T‐1131>C variation in the APOAV gene affects the plasma triglyceride showing a higher level in C‐1131 carriers than in T/T‐1131 homozygotes. This association has been observed both in males and females (p < 0.001). Similarly, plasma triglycerides were also significantly influenced by the S19>W APOAV genotypes. In both males and females, the W19 carriers have triglycerides significantly (p < 0.001) higher compared to the S19 homozygotes. In a group of MI patients, the frequency of the rare homozygotes for at least one APOAV polymorphism (C/C‐1131 and/or W/W19) was significantly higher than that in the population sample (7.4 vs 2.0%, p < 0.00001). We conclude that variations in the APOAV gene not only play a role in genetic determination of triglyceride levels but also could influence risk of MI.