bluuebunny/crossref_metadata_2025_split huggingface dataset:
Time-resolved luminescence resonance energy transfer imaging of protein–protein interactions in living cells
Authors: Rajapakse Harsha E. , Gahlaut Nivriti , Mohandessi Shabnam , Yu Dan , Turner Jerrold R. , Miller Lawrence W. 
Published: July 2010

Förster resonance energy transfer (FRET) with fluorescent proteins permits high spatial resolution imaging of protein–protein interactions in living cells. However, substantial non-FRET fluorescence background can obscure small FRET signals, making many potential interactions unobservable by conventional FRET techniques. Here we demonstrate time-resolved microscopy of luminescence resonance energy transfer (LRET) for live-cell imaging of protein–protein interactions. A luminescent terbium complex, TMP-Lumi4, was introduced into cultured cells using two methods: (
i
) osmotic lysis of pinocytic vesicles; and (
ii
) reversible membrane permeabilization with streptolysin O. Upon intracellular delivery, the complex was observed to bind specifically and stably to transgenically expressed
Escherichia coli
dihydrofolate reductase (eDHFR) fusion proteins. LRET between the eDHFR-bound terbium complex and green fluorescent protein (GFP) was detected as long-lifetime, sensitized GFP emission. Background signals from cellular autofluorescence and directly excited GFP fluorescence were effectively eliminated by imposing a time delay (10 μs) between excitation and detection. Background elimination made it possible to detect interactions between the first PDZ domain of ZO-1 (fused to eDHFR) and the C-terminal YV motif of claudin-1 (fused to GFP) in single microscope images at subsecond time scales. We observed a highly significant (
P
< 10
-6
), six-fold difference between the mean, donor-normalized LRET signal from cells expressing interacting fusion proteins and from control cells expressing noninteracting mutants. The results show that time-resolved LRET microscopy with a selectively targeted, luminescent terbium protein label affords improved speed and sensitivity over conventional FRET methods for a variety of live-cell imaging and screening applications.
Combining metabolic and protein engineering of a terpenoid biosynthetic pathway for overproduction and selectivity control
Authors: Leonard Effendi , Ajikumar Parayil Kumaran , Thayer Kelly , Xiao Wen-Hai , Mo Jeffrey D. , Tidor Bruce , Stephanopoulos Gregory , Prather Kristala L. J. 
Published: July 2010

A common strategy of metabolic engineering is to increase the endogenous supply of precursor metabolites to improve pathway productivity. The ability to further enhance heterologous production of a desired compound may be limited by the inherent capacity of the imported pathway to accommodate high precursor supply. Here, we present engineered diterpenoid biosynthesis as a case where insufficient downstream pathway capacity limits high-level levopimaradiene production in
Escherichia coli
. To increase levopimaradiene synthesis, we amplified the flux toward isopentenyl diphosphate and dimethylallyl diphosphate precursors and reprogrammed the rate-limiting downstream pathway by generating combinatorial mutations in geranylgeranyl diphosphate synthase and levopimaradiene synthase. The mutant library contained pathway variants that not only increased diterpenoid production but also tuned the selectivity toward levopimaradiene. The most productive pathway, combining precursor flux amplification and mutant synthases, conferred approximately 2,600-fold increase in levopimaradiene levels. A maximum titer of approximately 700 mg/L was subsequently obtained by cultivation in a bench-scale bioreactor. The present study highlights the importance of engineering proteins along with pathways as a key strategy in achieving microbial biosynthesis and overproduction of pharmaceutical and chemical products.
Catalyst-free synthesis of transparent, mesoporous diamond monoliths from periodic mesoporous carbon CMK-8
Authors: Zhang Li , Mohanty Paritosh , Coombs Neil , Fei Yingwei , Mao Ho-Kwang , Landskron Kai 
Published: July 2010

We report on the synthesis of optically transparent, mesoporous, monolithic diamond from periodic mesoporous carbon CMK-8 at a pressure of 21 GPa. The phase transformation is already complete at a mild synthesis temperature of 1,300 °C without the need of a catalyst. Surprisingly, the diamond is obtained as a mesoporous material despite the extreme pressure. X-ray diffraction, SEM, transmission electron microscopy, selected area electron diffraction, high-resolution transmission electron microscopy, and Z-contrast experiments suggest that the mesoporous diamond is composed of interconnected diamond nanocrystals having diameters around 5–10 nm. The Brunauer Emmett Teller surface area was determined to be 33 m
2
g
-1
according Kr sorption data. The mesostructure is diminished yet still detectable when the diamond is produced from CMK-8 at 1,600 °C and 21 GPa. The temperature dependence of the porosity indicates that the mesoporous diamond exists metastable and withstands transformation into a dense form at a significant rate due to its high kinetic inertness at the mild synthesis temperature. The findings point toward ultrahard porous materials with potential as mechanically highly stable membranes.
Comprehensive analysis of karyotypic mosaicism between trophectoderm and inner cell mass
Authors: Johnson D.S. , Cinnioglu C. , Ross R. , Filby A. , Gemelos G. , Hill M. , Ryan A. , Smotrich D. , Rabinowitz M. , Murray M.J. 
Published: July 2010

Abstract
Aneuploidy has been well-documented in blastocyst embryos, but prior studies have been limited in scale and/or lack mechanistic data. We previously reported preclinical validation of microarray 24-chromosome preimplantation genetic screening in a 24-h protocol. The method diagnoses chromosome copy number, structural chromosome aberrations, parental source of aneuploidy and distinguishes certain meiotic from mitotic errors. In this study, our objective was to examine aneuploidy in human blastocysts and determine correspondence of karyotypes between trophectoderm (TE) and inner cell mass (ICM). We disaggregated 51 blastocysts from 17 couples into ICM and one or two TE fractions. The average maternal age was 31. Next, we ran 24-chromosome microarray molecular karyotyping on all of the samples, and then performed a retrospective analysis of the data. The average per-chromosome confidence was 99.95%. Approximately 80% of blastocysts were euploid. The majority of aneuploid embryos were simple aneuploid, i.e. one or two whole-chromosome imbalances. Structural chromosome aberrations, which are common in cleavage stage embryos, occurred in only three blastocysts (5.8%). All TE biopsies derived from the same embryos were concordant. Forty-nine of 51 (96.1%) ICM samples were concordant with TE biopsies derived from the same embryos. Discordance between TE and ICM occurred only in the two embryos with structural chromosome aberration. We conclude that TE karyotype is an excellent predictor of ICM karyotype. Discordance between TE and ICM occurred only in embryos with structural chromosome aberrations.
CaMsrB2, Pepper Methionine Sulfoxide Reductase B2, Is a Novel Defense Regulator against Oxidative Stress and Pathogen Attack
Authors: Oh Sang-Keun , Baek Kwang-Hyun , Seong Eun Soo , Joung Young Hee , Choi Gyung-Ja , Park Jeong Mee , Cho Hye Sun , Kim Eun Ah , Lee Sangku , Choi Doil 
Published: July 2010

AbstractReactive oxygen species (ROS) are inevitably generated in aerobic organisms as by-products of normal metabolism or as the result of defense and development. ROS readily oxidize methionine (Met) residues in proteins/peptides to form Met-R-sulfoxide or Met-S-sulfoxide, causing inactivation or malfunction of the proteins. A pepper (Capsicum annuum) methionine sulfoxide reductase B2 gene (CaMsrB2) was isolated, and its roles in plant defense were studied. CaMsrB2 was down-regulated upon inoculation with either incompatible or compatible pathogens. The down-regulation, however, was restored to the original expression levels only in a compatible interaction. Gain-of-function studies using tomato (Solanum lycopersicum) plants transformed with CaMsrB2 resulted in enhanced resistance to Phytophthora capsici and Phytophthora infestans. Inversely, loss-of-function studies of CaMsrB2 using virus-induced gene silencing in pepper plants (cv Early Calwonder-30R) resulted in accelerated cell death from an incompatible bacterial pathogen, Xanthomonas axonopodis pv vesicatoria (Xav) race 1, and enhanced susceptibility to a compatible bacterial pathogen, virulent X. axonopodis pv vesicatoria race 3. Measurement of ROS levels in CaMsrB2-silenced pepper plants revealed that suppression of CaMsrB2 increased the production of ROS, which in turn resulted in the acceleration of cell death via accumulation of ROS. In contrast, the CaMsrB2-transgenic tomato plants showed reduced production of hydrogen peroxide. Taken together, our results suggest that the plant MsrBs have novel functions in active defense against pathogens via the regulation of cell redox status.
Identification and characterization of a Vibrio mimicus gene encoding the heme/hemoglobin receptor
Authors: Tanabe Tomotaka , Funahashi Tatsuya , Moon Yong‐Hwa , Tamai Eiji , Yamamoto Shigeo 
Published: July 2010

ABSTRACTThe present authors have previously reported that Vibrio mimicus expresses 77‐kDa and 80‐kDa outer membrane proteins in response to iron‐limited conditions, and that the 77‐kDa protein serves as the receptor for ferriaerobactin. In this study, it was found that V. mimicus can use heme and hemoglobin as iron sources. FURTA was then applied to V. mimicus 7PT to obtain candidate gene fragments involved in utilization of heme and hemoglobin. One FURTA‐positive clone was shown to contain a partial gene, whose predicted amino acid sequence correlated with the N‐terminal amino acid sequence determined for the 80‐kDa outer membrane protein and also shared homology with heme/hemoglobin receptors of Gram‐negative bacteria. Based on this information, the entire gene (named mhuA), and a gene upstream of mhuA (named mhuB) encoding a LysR family of transcriptional activator, were cloned and analyzed. RNA analysis indicated that mhuA and mhuB are each transcribed from individual Fur‐regulated promoters. Moreover, RNA analysis of an mhuB deletion mutant and a promoter reporter assay coupled with β‐galactosidase suggested that MhuB could function as an activator for mhuA transcription. Finally, the role of MhuA as the heme/hemoglobin receptor was confirmed by construction of an mhuA deletion mutant and its complemented strain followed by growth assay.
Not all ants are equal: obligate acacia ants provide different levels of protection against mega‐herbivores
Authors: Martins Dino J. 
Published: July 2010

AbstractIn obligate ant–plant mutualisms, the asymmetric engagement of a single plant species with multiple ant species provides the opportunity for partners to vary in their behaviour. Variation in behaviour has implications for the interactions with third‐party species such as herbivores. This study assessed the effect of obligate ant‐mutualists (Crematogaster mimosae, Crematogaster nigriceps and Tetraponera penzigi) inhabiting the African ant‐acacia (Acacia drepanolobium) on three mega‐herbivore browsers: the Maasai giraffe (Giraffa camelopardalis tippelskirchi), the reticulated giraffe (Giraffa c. reticulata) and the black rhino (Diceros bicornis). Giraffes are abundant and wide‐ranging herbivores of the acacias, whereas black rhinos are localized and perennial herbivores of the acacias. Multiyear field studies comparing the ants’ aggressive behaviour and browsing by mega‐herbivores suggested differences between the tending abilities of the primary ant species inhabiting A. drepanolobium. Trees occupied by the aggressive ant species C. mimosae had significantly less browsing by giraffes and black rhino than trees occupied by other ant species. The results of this study provide evidence that ant‐mutualists on African acacias can serve as deterrents to mega‐herbivores and that different ant species vary in their tending abilities.
ORIGINAL ARTICLE: Optimal timing for the administration of intranasal dexmedetomidine for premedication in children
Authors: Yuen V. M. , Hui Theresa W. , Irwin M. G. , Yao T.‐J. , Wong G. L. , Yuen M. K. 
Published: July 2010

SummaryPrevious studies have shown that 1 μg.kg−1 intranasal dexmedetomidine produces significant sedation in children aged between 2 and 12 years. This investigation was designed to evaluate the onset time. One hundred children aged 1–12 years of ASA physical status 1–2 undergoing elective surgery were randomly allocated to five groups. Patients in groups A to D received intranasal dexmedetomidine 1 μg.kg−1. Patients in Group E received intranasal placebo (0.9% saline). Children from groups A, B, C, D and E had intravenous cannulation attempted at 30, 45, 60, 75 and 45 min respectively after intranasal drug or placebo administration. Vital signs, behaviour and sedation status of the children were assessed regularly until induction of anaesthesia. More children from groups A to D achieved satisfactory sedation at the time of cannulation when compared to group E (p < 0.001). The proportion of children who achieved satisfactory sedation was not significantly different among groups A to D. Overall 62% of the children who received intranasal dexmedetomidine had satisfactory sedation at the time of cannulation. The median (95% CI) time for onset of sedation was 25 (25–30) min. The median (95% CI) duration of sedation was 85 (55–100) min.
Can you shrinkwrap a cow? Protections available for the intellectual property of the animal breeding industry
Authors: Ogden E. R. , Weigel K. 
Published: July 2010

SummaryThere are currently four main intellectual property protection statutory schemes available: copyright, trade secret, trademark and patent. Each of these protects a different aspect of intellectual property, which leaves gaps of protection when an innovation does not fit squarely within the boundaries of the statutes. Contracts allow the industry to tailor the protection desired. One very common approach is to license the product via contract. Licences allow intellectual property owners to retain ownership and give permission to others to use the product. Although there are several types of licences, the most common is the field of use licence, which limits the licensee's use of the product. This often leads to price discrimination where various levels of restriction are offered at corresponding prices. The more rights retained by the owner, the more restricted the buyer is and the lower the purchase price allowing customers to choose the level of restriction they are willing to accept. Therefore, the different uses and needs of various customers can be accounted for and reflected in the price. The animal breeding industry is currently struggling to protect their innovations falling into these statutory gaps. The protection for animal breeding industry innovations is most likely through contract law rather than traditional intellectual property law. By taking advantage of the unique nature of contracts, industry will be able to tailor protection and pricing to best suit the variety of customers and uses for the products sold.
Genes affecting coat colour and pattern in domestic dogs: a review
Authors: Schmutz S. M. , Berryere T. G. 
Published: July 2010

SummaryTremendous progress has been made in identifying genes involved in pigmentation in dogs in the past few years. Comparative genomics has both aided and benefited from these findings. Seven genes that cause specific coat colours and/or patterns in dogs have been identified: melanocortin 1 receptor, tyrosinase related protein 1, agouti signal peptide, melanophilin, SILV (formerly PMEL17), microphthalmia‐associated transcription factor and beta‐defensin 103. Although not all alleles have been yet identified at each locus, DNA tests are available for many. The identification of these alleles has provided information on interactions in this complex set of genes involved in both pigmentation and neurological development. The review also discusses pleiotropic effects of some coat colour genes as they relate to disease. The alleles found in various breeds have shed light on some potential breed development histories and phylogenetic relationships. The information is of value to dog breeders who have selected for and against specific colours since breed standards and dog showing began in the late 1800s. Because coat colour is such a visible trait, this information will also be a valuable teaching resource.
The emotional labour of caring about looked‐after children
Authors: Leeson Caroline 
Published: July 2010

ABSTRACTQuality relationships form the backbone of social work with children and their families. They are particularly relevant in the close, intimate work with looked‐after children who have identified how important it is to them that their relationship with their social worker is positive, warm and meaningful. It is accepted that in order to achieve and maintain successful and meaningful relationships, practitioners need to engage at an emotional as well as a professional level. All too often this requires a trade‐off between organizational efficiency and the emotional work of caring for looked‐after children. Therefore, it would appear the role of corporate parent is increasingly difficult, involving complex decisions about how practitioners might best spend their time, where their loyalties lie and the quality and direction of the final output. Using data from a series of interviews with practitioners, this paper explores the difficulties of maintaining active emotional engagement with children using the sociological concept of emotional labour.
Use of HR‐NMR to classify propolis obtained using different harvesting methods
Authors: Papotti Giulia , Bertelli Davide , Plessi Maria , Rossi Maria Cecilia 
Published: July 2010

SummaryPropolis has various biological activities closely related to the composition which varies according to environmental factors and also to the method of production. This study was aimed at determining whether or not HR‐NMR and multivariate statistical analysis were able to classify propolis according to the method used to harvest it. Sixty propolis samples were analysed in all. The ethanolic propolis extracts were initially analysed for quantification of the main bioactive substances, balsams and waxes. The 1H NMR and heteronuclear multiple bond correlation spectra were then acquired. Spectral data were analysed by the application of multivariate statistical techniques (Factor Analysis and General Discriminant Analysis). The best results were obtained using the 1H NMR which furnishes a sufficiently effective model by analysing the spectral region between 4.50 and 13.00 ppm (predictive capacity: 96.7%).
Influence of pH, NaCl and pre‐incubation on utilisation of surimi wash water in generation of antioxidative material by using the Maillard reaction
Authors: Thongraung Chakree , Kangsanan Sureeporn 
Published: July 2010

SummaryMaillard reaction products (MRPs) were generated from reaction mixtures of surimi wash water (SWW) with glucose or fructose (5%w/v) heated at 95 °C for 2–12 h. The effects of pH, NaCl and pre‐incubation of SWW on the Maillard reaction and antioxidant capacity of MRPs were investigated. The antioxidative capacity of MRPs was determined by measuring free DPPH° radical scavenging activity and reducing power. The highest colour intensity (OD420) as well as antioxidative capacity was noted in the reaction mixture containing fructose at pH 9.0. The addition of NaCl (0.5–2.5%w/v) caused reduction in browning intensity but enhanced antioxidative capacity of the MRPs. Pre‐incubation of SWW at 45 °C for 4 h decreased soluble protein but increased the Maillard reaction and antioxidative capacity of MRPs. A positive effect of salt or pre‐incubation of SWW on the antioxidative capacity of MRPs was not associated with the soluble protein content in the reaction mixture.
Psychrophilic and psychrotrophic clostridia: sporulation and germination processes and their role in the spoilage of chilled, vacuum‐packaged beef, lamb and venison
Authors: Adam Katharine H. , Flint Steve H. , Brightwell Gale 
Published: July 2010

SummarySpoilage of beef, lamb and venison by psychrophilic and psychrotrophic clostridial species renders meat unacceptable resulting in financial losses and reduced consumer confidence. A number of clostridial strains, including Clostridium algidicarnis, Clostridium algidixylanolyticum, Clostridium estertheticum, Clostridium frigidicarnis and Clostridium gasigenes, have been implicated in red meat spoilage. Unlike other spoilers, these clostridia are able to grow in anaerobic conditions and at chilled temperatures (some at −1.5 °C the optimal storage temperature for chilled red meat). The spoilage they cause is characterised by softening of the meat, production of large amounts of drip (exudates), offensive odours and in the case of C. estertheticum and C. gasigenes production of gas. Spoilage occurs following the introduction of clostridial spores into vacuum packages during processing. Germination of spores is necessary for the growth of vegetative cells, which cause spoilage. Current mitigation strategies focus on good management practice within meat processing plants. However, this is not always sufficient to prevent spoilage. This review summarises the issues associated with meat spoilage because of psychrotolerant clostridia and discusses areas that require further study.
The capability of rosemary extract in preventing oxidation of fish lipid
Authors: Ozogul Yesim , Ayas Deniz , Yazgan Hatice , Ozogul Fatih , Boga Esmeray K. , Ozyurt Gulsun 
Published: July 2010

SummaryThe effects of rosemary extract at different levels (%1, R1, and %2, R2) on the quality of vacuum‐packed sardine in terms of sensory, biochemical (thiobarbituric acid, total volatile basic nitrogen, peroxide value and free fatty acids) and microbiological analyses (total viable counts) were investigated. Fish were filleted and divided into three groups. First group was used as the control (C) without rosemary extract, second group was treated with 1% rosemary extracts (10 g L−1) for 2 min (R1), and the third was treated with 2% rosemary extracts (20 g L−1) for 2 min (R2). Thirty fillets per litre were used. After that, all groups were vacuum‐packed in polyethylene bags. The samples were stored in the refrigerator condition (4 ± 1 °C) over the storage period of 20 days. The results showed that the use of rosemary extract improved the sensory quality of both raw and cooked sardine, most preferably sardine treated with 1% of rosemary. Biochemical analysis showed that the use of 2% of rosemary extract were found to be most effective (P < 0.05) in controlling the rate of lipid oxidation.
Kinetics of ascorbic acid degradation and colour change in ground cashew apples treated at high temperatures (100–180 °C)
Authors: Lima Janice R. , Elizondo Nadiarid J. , Bohuon Philippe 
Published: July 2010

SummaryAscorbic acid (AA) degradation and colour changes, measured by the lightness index (L*), were determined in cashew apples (at low dissolved O2 concentrations) heated at high temperature (100–180 °C) in a hermetically sealed cell. A nonisothermal method was developed to estimate thermal degradation kinetics. The results showed that reaction kinetics during heat treatments were well represented by first‐order reactions. The temperature dependence of the kinetic constants was described by an Arrhenius type equation. The activation energy (Ea) for AA degradation and lightness index were 94 ± 3 and 98 ± 3 kJ mol−1, respectively. The reaction rate constant at 140 °C for AA degradation (64 × 10−5 ± 3 × 10−5 s−1) was twice that for the lightness index change (33 × 10−5 ± 2 × 10−5 s−1). Results allow generating temperature profiles of heat processes that would help preserve the AA of cashew apples as well as control the colour formation during high‐temperature processes.