Poly-(γ-glutamic acid) Production and Optimization from Agro-Industrial Bioresources as Renewable Substrates by Bacillus sp. FBL-2 through Response Surface Methodology.
We optimized culture conditions using Bacillus sp. FBL-2 as a poly-(γ-glutamic acid) (PGA) producing strain isolated from cheonggukjang. All experiments were performed under aerobic conditions using a laboratory scale 2.5 L fermentor. We investigated the effects of fermentation parameters (temperature, pH, agitation, and aeration) and medium components (glutamic acid, citric acid, and yeast extract) on poly-(γ-glutamic acid) production, viscosity, and dry cell mass. A non-optimized fermentation method (1.5 vvm, 350 rpm, and 37 °C) yielded PGA, viscosity, and dry cell mass at levels of 100.7 g/L, 483.2 cP, and 3.4 g/L, respectively. L-glutamic acid, citric acid, and yeast extract supplementation enhanced poly-(γ-glutamic acid) production to 175.9 g/L. Additionally, the production of poly-(γ-glutamic acid) from rice bran and wheat bran was assessed using response surface methodology (central composite rotatable design). Agricultural byproducts (rice bran and wheat bran) and H2SO4 were selected as factors, and experiments were performed by combining various component concentrations to determine optimal component concentrations. Our experimentally-derived optimal parameters included 38.6 g/L of rice bran, 0.42% of H2SO4, 28.0 g/L of wheat bran, and 0.32% of H2SO4. Under optimum conditions, rice bran medium facilitated poly-(γ-glutamic acid) production of up to 22.64 g/L, and the use of wheat bran medium yielded up to 14.6 g/L. Based on a validity test using the optimized culture conditions, poly-(γ-glutamic acid) was produced at 47.6 g/L and 36.4 g/L from these respective mediums, and both results were higher than statistically predicted. This study suggests that rice bran can be used as a potential alternative substrate for poly-(γ-glutamic acid) production.


Introgression study reveals two quantitative trait loci involved in interspecific variation in memory retention among Nasonia wasp species.
Genes involved in the process of memory formation have been studied intensively in model organisms; however, little is known about the mechanisms that are responsible for natural variation in memory dynamics. There is substantial variation in memory retention among closely related species in the parasitic wasp genus Nasonia. After a single olfactory conditioning trial, N. vitripennis consolidates long-term memory that lasts at least 6 days. Memory of the closely related species N. giraulti is present at 24 h but is lost within 2 days after a single trial. The genetic basis of this interspecific difference in memory retention was studied in a backcrossing experiment in which the phenotype of N. giraulti was selected for in the background of N. vitripennis for up to five generations. A genotyping microarray revealed five regions that were retained in wasps with decreased memory retention. Independent introgressions of individual candidate regions were created using linked molecular markers and tested for memory retention. One region on chromosome 1 (spanning ∼5.8 cM) and another on chromosome 5 (spanning ∼25.6 cM) resulted in decreased memory after 72 h, without affecting 24-h-memory retention. This phenotype was observed in both heterozygous and homozygous individuals. Transcription factor CCAAT/enhancer-binding protein and a dopamine receptor, both with a known function in memory formation, are within these genomic regions and are candidates for the regulation of memory retention. Concluding, this study demonstrates a powerful approach to study variation in memory retention and provides a basis for future research on its genetic basis.


Dosimetric evaluation of radionuclides for VCAM-1-targeted radionuclide therapy of early brain metastases.
Brain metastases develop frequently in patients with breast cancer, and present a pressing therapeutic challenge. Expression of vascular cell adhesion molecule 1 (VCAM-1) is upregulated on brain endothelial cells during the early stages of metastasis and provides a target for the detection and treatment of early brain metastases. The aim of this study was to use a model of early brain metastasis to evaluate the efficacy of α-emitting radionuclides, 149Tb, 211At, 212Pb, 213Bi and 225Ac; β-emitting radionuclides, 90Y, 161Tb and 177Lu; and Auger electron (AE)-emitters 67Ga, 89Zr, 111In and 124I, for targeted radionuclide therapy (TRT). Histologic sections and two photon microscopy of mouse brain parenchyma were used to inform a cylindrical vessel geometry using the Geant4 general purpose Monte Carlo (MC) toolkit with the Geant4-DNA low energy physics models. Energy deposition was evaluated as a radial function and the resulting phase spaces were superimposed on a DNA model to estimate double-strand break (DSB) yields for representative β- and α-emitters, 177Lu and 212Pb. Relative biological effectiveness (RBE) values were determined by only evaluating DNA damage due to physical interactions. 177Lu produced 2.69 ± 0.08 DSB per GbpGy, without significant variation from the lumen of the vessel to a radius of 100 µm. The DSB yield of 212Pb included two local maxima produced by the 6.1 MeV and 8.8 MeV α-emissions from decay products, 212Bi and 212Po, with yields of 7.64 ± 0.12 and 9.15 ± 0.24 per GbpGy, respectively. Given its higher DSB yield 212Pb may be more effective for short range targeting of early micrometastatic lesions than 177Lu. MC simulation of a model of early brain metastases provides invaluable insight into the potential efficacy of α-, β- and AE-emitting radionuclides for TRT. 212Pb, which has the attributes of a theranostic radionuclide since it can be used for SPECT imaging, showed a favorable dose profile and RBE.


Local effects drive heterozygosity-fitness correlations in an outcrossing long-lived tree.
Heterozygosity-fitness correlations (HFCs) have been used to understand the complex interactions between inbreeding, genetic diversity and evolution. Although frequently reported for decades, evidence for HFCs was often based on underpowered studies or inappropriate methods, and hence their underlying mechanisms are still under debate. Here, we used 6100 genome-wide single nucleotide polymorphisms (SNPs) to test for general and local effect HFCs in maritime pine (Pinus pinaster Ait.), an iconic Mediterranean forest tree. Survival was used as a fitness proxy, and HFCs were assessed at a four-site common garden under contrasting environmental conditions (total of 16 288 trees). We found no significant correlations between genome-wide heterozygosity and fitness at any location, despite variation in inbreeding explaining a substantial proportion of the total variance for survival. However, four SNPs (including two non-synonymous mutations) were involved in significant associations with survival, in particular in the common gardens with higher environmental stress, as shown by a novel heterozygosity-fitness association test at the species-wide level. Fitness effects of SNPs involved in significant HFCs were stable across maritime pine gene pools naturally growing in distinct environments. These results led us to dismiss the general effect hypothesis and suggested a significant role of heterozygosity in specific candidate genes for increasing fitness in maritime pine. Our study highlights the importance of considering the species evolutionary and demographic history and different spatial scales and testing environments when assessing and interpreting HFCs.


Three different fibronectin mRNAs arise by alternative splicing within the coding region.
We report the isolation of cDNA clones for fibronectin from a rat liver library prepared in the expression vector, lambda gt11. Restriction mapping and DNA sequencing of these clones establish the sequence of the C-terminal 35% of rat fibronectin, covering the cell-, heparin-, and fibrin-binding domains. The cell- and heparin-binding regions have homologous repeating sequences. Based on the sequence data and S1 nuclease mapping, we conclude that there are at least three different fibronectin mRNAs in rat liver which differ in coding potential. The three RNAs appear to arise by alternative splicing within the coding region and are probably all encoded by a single gene. The implications of these results for the structure and function of fibronectin and the differences between various types of fibronectin are discussed.


Conservation of a core neurite transcriptome across neuronal types and species.
The intracellular localization of mRNAs allows neurons to control gene expression in neurite extensions (axons and dendrites) and respond rapidly to local stimuli. This plays an important role in diverse processes including neuronal growth and synaptic plasticity, which in turn serves as a foundation for learning and memory. Recent high-throughput analyses have revealed that neurites contain hundreds to thousands of mRNAs, but an analysis comparing the transcriptomes derived from these studies has been lacking. Here we analyze 20 datasets pertaining to neuronal mRNA localization across species and neuronal types and identify a conserved set of mRNAs that had robustly localized to neurites in a high number of the studies. The set includes mRNAs encoding for ribosomal proteins and other components of the translation machinery, mitochondrial proteins, cytoskeletal components, and proteins associated with neurite formation. Our combinatorial analysis provides a unique resource for future hypothesis-driven research. This article is categorized under: RNA Export and Localization > RNA Localization RNA Evolution and Genomics > Computational Analyses of RNA RNA Methods > RNA Analyses in Cells.


Reamed intramedullary femoral nailing after induction of an "ARDS-like" state in sheep: effect on clinically applicable markers of pulmonary function.
At present, the optimal treatment for appropriately resuscitated, multiply injured patients includes fixation of long bone fractures within twenty-four hours of injury. This management approach has been shown to decrease the incidence of pulmonary complications, multiple organ failure, and death. Some investigators have hypothesized that acute reamed intramedullary nailing of the femur (RIMNF) may result in pulmonary dysfunction as a result of the pulmonary fat embolization generated during this procedure. Patients with concomitant thoracic trauma may be at particular risk for this potentially severe complication. In an attempt to determine whether RIMNF can be safely carried out regardless of the severity of a pulmonary injury, we monitored the pulmonary effects of RIMNF in sheep in which an acute respiratory disorder (ARDS)-like state had been induced. Our hypothesis was that, if the pulmonary fat embolization that occurs as a result of RIMNF has a clinically significant effect, it would be detectable in an animal model in which a severe lung injury had been induced prior to the start of RIMNF. This was an acute experimental procedure performed on yearling sheep. Reamed intramedullary nailing of the femur was performed in two groups of instrumented sheep. The first group had no pulmonary injuries. The second group had an ARDS-like state induced by intravenous infusion of perilla ketone prior to RIMNF. Perilla ketone increases pulmonary microvascular permeability without changing filling pressures and is used to induce a model of human ARDS. Hemodynamic and oximetric parameters were measured or calculated, as was pulmonary dynamic compliance during the experiment. Infusion of perilla ketone caused a significant pulmonary injury. RIMNF caused no additional significant effect on intrapulmonary shunt, mixed venous oxygen saturation, or dynamic compliance, which are clinically used to assess the severity of pulmonary dysfunction in injured patients. The fat embolization that occurs during RIMNF in an appropriately resuscitated sheep has no clinically significant effect on pulmonary function, even in the setting of a severe pulmonary dysfunction.


Hard science versus phenomenology in reproductive immunology.
It has been suggested that studies of immunological aspects of reproduction have recently changed from the phenomenological (e.g., speculative/descriptive) to hard science (e.g., precise molecular description/ explanation). The significance of this development is explored by analysis of the contribution made by hard science that has led to a number of assertions. Does HLA-G determine maternal tolerance of her semi-allogeneic fetus? Do differences in placentation between humans, rodents, and other species make direct comparisons largely meaningless? Does membrane cofactor protein (CD46) contribute to pregnancy success by protecting sperm and fetal trophoblast from complement-mediated lysis? Does a low frequency in mice of maternal T cells specific for paternal alloantigens occur, and if so, does phenomenon explain specific maternal tolerance in pregnancy? Do placental and decidual macrophage components provide an important immediate antigen-nonspecific host defense to infection? Does the mix of bioactive molecules make the uterus an immunologically privileged site, and does the molecular melange have a pivotal role in promoting growth and development of the placenta and embryo? Does prolactin acting on receptors on lymphocytes suppress function and thereby account for remission of autoimmune disease in pregnant women? Is the efficacy and safety of immunotherapy for recurrent spontaneous abortion a thorny issue that will be resolved by an ongoing funded national trial? What is a scientifically correct timeline of important discoveries and developments in reproductive immunology? It is argued that it is unprofitable to divorce phenomenology from hard science and that the timeline for developments in reproductive immunology begin with Darwin. Irrespective of whether one is dealing with phenomena or hard science data, Hippocrates was correct in his aphorism that "description is infinite and easy; explanation is limited and difficult".


Regulatory circuits controlling white versus brown adipocyte differentiation.
Adipose tissue is a major endocrine organ that exerts a profound influence on whole-body homoeostasis. Two types of adipose tissue exist in mammals: WAT (white adipose tissue) and BAT (brown adipose tissue). WAT stores energy and is the largest energy reserve in mammals, whereas BAT, expressing UCP1 (uncoupling protein 1), can dissipate energy through adaptive thermogenesis. In rodents, ample evidence supports BAT as an organ counteracting obesity, whereas less is known about the presence and significance of BAT in humans. Despite the different functions of white and brown adipocytes, knowledge of factors differentially influencing the formation of white and brown fat cells is sparse. Here we summarize recent progress in the molecular understanding of white versus brown adipocyte differentiation, including novel insights into transcriptional and signal transduction pathways. Since expression of UCP1 is the hallmark of BAT and a key factor determining energy expenditure, we also review conditions associated with enhanced energy expenditure and UCP1 expression in WAT that may provide information on processes involved in brown adipocyte differentiation.


Qualitative investigation of the factors that generate ambivalent feelings in women who give birth after receiving negative results from non-invasive prenatal testing.
Women who receive negative results from non-invasive prenatal genetic testing (NIPT) may find that they later have mixed or ambivalent feelings, for example, feelings of accepting NIPT and regretting undergoing the test. This study aimed to investigate the factors generating ambivalent feelings among women who gave birth after having received negative results from NIPT. A questionnaire was sent to women who received a negative NIPT result, and a contents analysis was conducted focusing on ambivalent expressions for those 1562 women who responded the questionnaire. The qualitative data gathered from the questionnaire were analyzed using the N-Vivo software package. Environmental factors, genetic counseling-related factors, and increased anticipatory anxiety, affected the feeling of ambivalence among pregnant women. Furthermore, pregnant women desired more information regarding the detailed prognosis for individuals with Down syndrome and living with them and/or termination, assuming the possibility that they were positive. Three major interrelated factors affected the feeling of ambivalence in women. Highlighting and discussing such factors during genetic counseling may resolve some of these ambivalences, thereby enhancing the quality of decisions made by pregnant women.


BHK21 fibroblast aggregation inhibited by glycopeptides from the cell surface.
Glycopeptides were removed by trypsinization from the surface of baby hamster kidney cells (line BHK21-C13), digested by pronase and separated into 2 fractions by exclusion chromatography. The addition of small amounts of either glycopeptide fraction to shaken suspensions of lightly trypsinzied cells inhibited their rapid aggregation, but one fraction was more active than the other and in higher concentrations it was able to inhibit aggregation completely. After this fraction was purified by high-voltage electrophoresis one subfraction also inhibited aggregation. The effect of the glycopeptides increased following their pretreatment with neuraminidase, but preincubation with periodiate or galactose oxidase destroyed all activity. Galactose oxidase also inhibited cell aggregation directly. Similar glycopeptides from virus-transformed BHK21 cells, oligosaccharides and intact and desialysed human urinary glycoproteins had comparatively little or no effect on BHK21 cell aggregation. The results suggest terminal beta-galactosides and possible alpha-galactosides, and to some extent a particular substructure of cell surface heteroglycans are necessary for their inhibitory activity. The parent, plasma membrane of glycoproteins might serve as adhesive binding sites in cell cohesion, but some evidence indicates cell surface sialyl- and galactosyltransferases may not ordinarily act as their complementary binding receptors.


Multidrug-resistant Salmonella enterica serovar paratyphi A harbors IncHI1 plasmids similar to those found in serovar typhi.
Salmonella enterica serovars Typhi and Paratyphi A cause systemic infections in humans which are referred to as enteric fever. Multidrug-resistant (MDR) serovar Typhi isolates emerged in the 1980s, and in recent years MDR serovar Paratyphi A infections have become established as a significant problem across Asia. MDR in serovar Typhi is almost invariably associated with IncHI1 plasmids, but the genetic basis of MDR in serovar Paratyphi A has remained predominantly undefined. The DNA sequence of an IncHI1 plasmid, pAKU_1, encoding MDR in a serovar Paratyphi A strain has been determined. Significantly, this plasmid shares a common IncHI1-associated DNA backbone with the serovar Typhi plasmid pHCM1 and an S. enterica serovar Typhimurium plasmid pR27. Plasmids pAKU_1 and pHCM1 share 14 antibiotic resistance genes encoded within similar mobile elements, which appear to form a 24-kb composite transposon that has transferred as a single unit into different positions into their IncHI1 backbones. Thus, these plasmids have acquired similar antibiotic resistance genes independently via the horizontal transfer of mobile DNA elements. Furthermore, two IncHI1 plasmids from a Vietnamese isolate of serovar Typhi were found to contain features of the backbone sequence of pAKU_1 rather than pHCM1, with the composite transposon inserted in the same location as in the pAKU_1 sequence. Our data show that these serovar Typhi and Paratyphi A IncHI1 plasmids share highly conserved core DNA and have acquired similar mobile elements encoding antibiotic resistance genes in past decades.


Insurance Coverage and Treatment Use Under the Affordable Care Act Among Adults With Mental and Substance Use Disorders.
Many adults who have mental or substance use disorders or both experience insurance-related barriers to care, contributing to low treatment utilization. Expanded insurance under the Affordable Care Act (ACA) could improve coverage and access. The study identified changes in coverage and treatment use following 2014 ACA insurance expansions. Data from the National Survey on Drug Use and Health were used to identify individuals ages 18-64 screening positive for any mental disorder (N=29,962) or substance use disorder (N=19,243) for two periods: 2011-2013 and 2014. Regression-adjusted means were calculated for insurance rates and treatment used in each period overall and among individuals with household incomes ≤200% of the federal poverty level (FPL). Compared with 2011-2013, in 2014 significant reductions were seen in the uninsured rate for individuals with mental disorders (-5.4 percentage points, p<.01) and substance use disorders (-5.1 percentage points, p<.01). Increases in insurance coverage occurred mostly through Medicaid. Insurance gains were larger for adults with incomes ≤200% of FPL compared with the overall sample. Use of mental health treatment increased by 2.1 percentage points (p=.04), but use of substance use disorder treatment did not change. No significant changes were noted in treatment settings for mental and substance use disorder treatments. Payment by Medicaid for substance use disorder treatment increased by 7.4 percentage points (p=.05). Sizable increases in coverage for adults with mental disorders and adults with substance use disorders were identified in the year following the 2014 ACA expansions; however, low treatment rates among this population remain a concern. Initiatives to engage the newly insured in treatment are needed.


Small proportions of actively-smoking patrons and high PM2.5 levels in southern California tribal casinos: support for smoking bans or designated smoking areas.
Nearly all California casinos currently allow smoking, which leads to potentially high patron exposure to secondhand tobacco smoke pollutants. Some argue that smoking restrictions or bans would result in a business drop, assuming > 50% of patrons smoke. Evidence in Nevada and responses from the 2008 California tobacco survey refute this assertion. The present study investigates the proportion of active smokers in southern California tribal casinos, as well as occupancy and PM(2.5) levels in smoking and nonsmoking sections. We measured active-smoker and total-patron counts during Friday or Saturday night visits (two per casino) to smoking and nonsmoking gaming areas inside 11 southern California casinos. We counted slot machines and table games in each section, deriving theoretical maximum capacities and occupancy rates. We also measured PM(2.5) concentrations (or used published levels) in both nonsmoking and smoking areas. Excluding one casino visit with extremely high occupancy, we counted 24,970 patrons during 21 casino visits of whom 1,737 were actively smoking, for an overall active- smoker proportion of 7.0% and a small range of ~5% across casino visits (minimum of 5% and maximum of 10%). The differences in mean inter-casino active-smoker proportions were not statistically significant. Derived occupancy rates were 24% to 215% in the main (low-stakes) smoking-allowed slot or table areas. No relationship was found between observed active-smoker proportions and occupancy rate. The derived maximum capacities of nonsmoking areas were 1% to 29% of the overall casino capacity (most under 10%) and their observed occupancies were 0.1 to over 3 times that of the main smoking-allowed casino areas. Seven of twelve visits to nonsmoking areas with no separation had occupancy rates greater than main smoking areas. Unenclosed nonsmoking areas don't substantially protect occupants from PM2.5 exposure. Nonsmoking areas encapsulated inside smoking areas or in a separate, but unenclosed, area had PM(2.5) levels that were 10 to 60 μg/m(3) and 6 to 23 μg/m(3) higher than outdoor levels, respectively, indicating contamination from smoking. Although fewer than roughly 10% of casino patrons are actively smoking on average, these individuals substantially increase PM(2.5) exposure for all patrons in smoking and unenclosed nonsmoking areas. Nonsmoking areas may be too inconvenient, small, or undesirable to serve a substantial number of nonsmoking patrons. Imposing indoor smoking bans, or contained smoking areas with a maximum capacity of up to 10% of the total patronage, would offer protection from PM2.5 exposures for nonsmoking patrons and reduce employee exposures.


A microdomain for protein secretion in Gram-positive bacteria.
Gram-positive bacteria face unique challenges in generating biologically active conformations for their exported proteins because they lack a dedicated compartment for folding secreted polypeptides. We have discovered that protein secretion by way of the general secretory (Sec) pathway in the important human pathogen Streptococcus pyogenes proceeds through a single microdomain. Unlike other mechanisms for asymmetry involving the Sec pathway, proteins destined for secretion are targeted to a single locus distal to either cell pole that has specialized to contain the Sec translocons. This subcellular organization may represent a paradigm for secretion common to other Gram-positive pathogens with profound implications for pathogenesis.


Secondary cytoreductive surgery for patients with relapsed epithelial ovarian carcinoma: who benefits?
This study was performed to address patient selection criteria and the role of secondary cytoreductive surgery (SCR) in patients with epithelial ovarian carcinoma (EOC) who had relapsed tumors after a progression-free interval > or = 3 months. One hundred seventeen patients with relapsed EOC after a clinical complete remission duration > or = 3 months who underwent SCR were entered on this prospective trial. Survival curves were generated using the Kaplan-Meier method, and statistical comparisons were performed using log-rank tests, logistic stepwise regression analyses, and a Cox stepwise regression model. The median patient age at the time of relapse was 53 years (range, 20-78 years). The median survival was 22 months and the estimated 5-year survival rate for the entire cohort was 17.2%. Tumor was confined to a solitary site in 33 patients and to > or = 2 sites in 84 patients. After they underwent SCR, 11 patients were rendered macroscopically disease free, 61 patients had residual disease that measured < or = 1 cm in greatest dimension, and 45 patients had bulky intraabdominal residual disease. Survival was influenced by the extent of relapse disease (solitary site vs. multiple sites; P < 0.0001), the size of residual disease after SCR (0 cm vs. < or = 1 cm [P = 0.1211], < or = 1 cm vs. > 1 cm [P = 0.0002], and 0 cm vs. > 1 cm [P = 0.0011]), Eastern Cooperative Oncology Group performance status (0 vs. 1 [P = 0.134], 1 vs. 2 [P = 0.007], and 0 vs. 2 [P = 0.0012]), and the number of cycles of salvage chemotherapy (1-2 cycles vs. 3-5 cycles [P = 0.0144]; 1-2 cycles vs. > or = 6 cycles [P < 0.0001]; and 3-5 cycles vs. > or = 6 cycles [P = 0.0009]). The outcome of SCR was influenced by the extent of relapse disease (multiple sites [51.2%] vs. solitary sites [87.9%]; relative risk [RR] = 9.1237; P = 0.0002) and by the use of bowel resection (yes [60.9%] vs. no [37.5%]; RR = 0.3828; P = 0.0106). SCR was found to be safe for patients with relapsed EOC who achieved a clinical complete remission that lasted > or = 3 months, with resectability similar to that of primary debulking surgery. Optimal surgical outcomes were achieved easily in patients who apparently had solitary tumor sites, with bowel resection making it possible to remove bulky tumors that involved the intestine. A survival benefit was provided by optimal SCR, particularly when surgery was supported by multiple courses of salvage chemotherapy.


Context-specific ontology integration: a bayesian approach.
We introduce a principled computational framework and methodology for automated discovery of context-specific functional links between ontologies. Our model leverages over disparate free-text literature resources to score the model of dependency linking two terms under a context against their model of independence. We identify linked terms as those having a significant bayes factor (p < 0.01). To scale our algorithm over massive ontologies, we propose a heuristic pruning technique as an efficient algorithm for inferring such links.We have applied this method to translationalize Gene Ontology to all other ontologies available at National Center of Biomedical Ontology (NCBO) BioPortal under the context of Human Disease ontology. Our results show that in addition to broadening the scope of hypothesis for researchers, our work can potentially be used to explore continuum of relationships among ontologies to guide various biological experiments.


A Focus Group Study on African American Living Donors' Treatment Preferences, Sociocultural Factors, and Health Beliefs About Apolipoprotein L1 Genetic Testing.
Because apolipoprotein L1 (APOL1) risk variants may contribute to live donors' kidney failure postdonation, professional guidelines suggest informing potential donors with African ancestry about the availability of APOL1 genotyping. This study assessed African American (AA) donors' perceptions of APOL1 genetic testing and how APOL1 may affect ethnic identity. Four focus groups were conducted with AA donors about their decision-making for and perceptions of APOL1 genetic testing and donation to inform a new culturally targeted educational brochure on APOL1 genetic testing. Qualitative data were analyzed by thematic analysis. Seventeen donors participated (47% participation rate). Four major themes emerged. (1) In hypothetical scenarios, most participants would have undergone APOL1 testing during donor evaluation to make a more informed decision, but many would have still donated. (2) Participants desired information about how having 2 APOL1 risk variants affects the donor's and the recipient's health. (3) Participants referred to diversity of genetic ancestry and cultural constructions of racial/ethnic identity to question the population at risk for APOL1 risk variants and recommended that all potential donors undergo genetic testing and receive education about APOL1. (4) Participants worried that out-of-pocket costs would deter APOL1 testing and that APOL1 could become a preexisting condition and discriminate against AAs. Our findings suggest that AA donors desire APOL1 testing to foster informed consent. Transplant clinicians should be aware of these responses to APOL1 testing and be sensitive to historical issues of distrust and discrimination.


The Antioxidant and Safety Properties of Spent Coffee Ground Extracts Impacted by the Combined Hot Pressurized Liquid Extraction-Resin Purification Process.
Hot pressurized liquid extraction has been used to obtain polyphenols; however, its operating conditions can generate hydroxymethylfurfural, a potential human carcinogen. The addition of ethanol can reduce process temperatures and retain extraction efficiencies, but the ethanol may reduce the recovery of polyphenols in the subsequent purification stage, affecting the antioxidant properties of the extracts. This study evaluates a combined hot pressurized liquid extraction-resin purification process to obtain polyphenol extracts from spent ground coffee reduced in hydroxymethylfurfural. A multifactorial design was developed to determine the combined effect of the extraction (ethanol content: 0-16% and temperature: 60-90 °C) and purification (ethanol: 60-80%) conditions on some chemical properties of the extracts. The highest recovery of polyphenols (~8 mg GAE/g dry coffee solids) and reduction of hydroxymethylfurfural (95%) were obtained at 90 °C and 16% of ethanol during extraction and 80% of ethanol during purification. These operating conditions retained the antioxidant capacity of the crude extract between 60% and 88% depending on the determination method and recovered 90, 98, and 100% of 4-feruloylquinic acid, epicatechin, and 5-feruloylquinic acid, respectively after purification. The combined process allows differential polyphenols' recovery and enhances the safety of the extracts. Our computational chemistry results ruled out that the overall selectivity of the integrated process was correlated with the size of the polyphenols.


Melatonin and lipid uptake by murine fibroblasts: clinical implications.
The current study was undertaken to uncover the role of melatonin in lipid metabolism in the murine fibroblasts. The results show melatonin in vitro enhances lipid accumulation and lipid droplet formation in this cell line. Using oil red O staining, it was found that when oleic acid was present in the culture media, melatonin at doses of 0.1-2mM, significantly increased the lipid concentrations in the cells. However, low levels of melatonin, with or without oleic acid, did not influence lipid metabolism in the cultured fibroblasts. When a non-specific melatonin receptor antagonist, luzindole 10 microM was co-incubated with 1mM melatonin, the stimulatory effects of melatonin on lipid accumulation in these cells was significantly reduced. It appears that the effects of melatonin on lipid metabolism in murine fibroblasts is mediated by melatonin membrane receptors.


Complete genome of the marine bacterium Wenzhouxiangella marina KCTC 42284(T).
Wenzhouxiangella marina is an obligatory aerobic, Gram-negative, non-motile, rod-shaped bacterium that was isolated from the culture broth of marine microalgae, Picochlorum sp. 122. Here we report the 3.67 MB complete genome (65.26 G+C%) of W. marina KCTC 42284(T) encoding 3,016 protein-coding genes, 43 tRNAs and one rRNA operon. The genomic information supports multiple horizontal gene transfer (HGT) events in the history of W. marina, possibly with other marine bacteria co-existing in marine habitats. Evaluation of genomic signatures revealed 19 such HGT-derived genomic islands. Of these, eight were also supported by "genomic context" that refers to the existence of integrases, transposases and tmRNA genes either inside or in near vicinity to the island. The addition of W. marina genome expands the repertoire of marine bacterial genomic diversity, especially because the strain represents the sole genomic resource of a novel taxonomic family in the bacterial order Chromatiales.


A Journey in Diabetes: From Clinical Physiology to Novel Therapeutics: The 2020 Banting Medal for Scientific Achievement Lecture.
Insulin resistance and β-cell dysfunction are the core pathophysiological mechanisms of all hyperglycemic syndromes. Advances in in vivo investigative techniques have made it possible to quantify insulin resistance in multiple sites (skeletal and myocardial muscle, subcutaneous and visceral fat depots, liver, kidney, vascular tissues, brain and intestine), to clarify its consequences for tissue substrate selection, and to establish its relation to tissue perfusion. Physiological modeling of β-cell function has provided a uniform tool to measure β-cell glucose sensitivity and potentiation in response to a variety of secretory stimuli, thereby allowing us to establish feedbacks with insulin resistance, to delineate the biphasic time course of conversion to diabetes, to gauge incretin effects, and to identify primary insulin hypersecretion. As insulin resistance also characterizes several of the comorbidities of diabetes (e.g., obesity, hypertension, dyslipidemia), with shared genetic and acquired influences, the concept is put forward that diabetes is a systemic disease from the outset, actually from the prediabetic stage. In fact, early multifactorial therapy, particularly with newer antihyperglycemic agents, has shown that the burden of micro- and macrovascular complications can be favorably modified despite the rising pressure imposed by protracted obesity.


Expression of growth-related genes in young and older human skeletal muscle following an acute stimulation of protein synthesis.
Muscle growth is associated with an activation of the mTOR signaling pathway and satellite cell regulators. The purpose of this study was to determine whether 17 selected genes associated with mTOR/muscle protein synthesis and the satellite cells/myogenic program are differentially expressed in young and older human skeletal muscle at rest and in response to a potent anabolic stimulus [resistance exercise + essential amino acid ingestion (RE+EAA)]. Twelve male subjects (6 young, 6 old) completed a bout of heavy resistance exercise. Muscle biopsies were obtained before and at 3 and 6 h post RE+EAA. Subjects ingested leucine-enriched essential amino acids at 1 h postexercise. mRNA expression was determined using qRT-PCR. At rest, hVps34 mRNA was elevated in the older subjects (P < 0.05) while there was a tendency for levels of myoD, myogenin, and TSC2 mRNA to be higher than young. The anabolic stimulus (RE+EAA) altered mRNAs associated with mTOR regulation. Notably, REDD2 decreased in both age groups (P < 0.05) but the expression of Rheb mRNA increased only in the young. Finally, cMyc mRNA was elevated (P < 0.05) in both young and old at 6 h post RE+EAA. Furthermore, RE+EAA also increased expression of several mRNAs associated with satellite function in the young (P < 0.05), while expression of these mRNAs did not change in the old. We conclude that several anabolic genes in muscle are more responsive in young men post RE+EAA. Our data provide new insights into the regulation of genes important for transcription and translation in young and old human skeletal muscle post RE+EAA.


Severe bereavement stress during the prenatal and childhood periods and risk of psychosis in later life: population based cohort study.
To examine the risk of psychosis associated with severe bereavement stress during the antenatal and postnatal period, between conception to adolescence, and with different causes of death. Population based cohort study. Swedish national registers including births between 1973 and 1985 and followed-up to 2006. In a cohort of 1,045,336 Swedish births (1973-85), offspring born to mothers exposed to severe maternal bereavement stress six months before conception or during pregnancy, or exposed to loss of a close family member subsequently from birth to 13 years of age were followed until 2006. Admissions were identified by linkage to national patient registers. Crude and adjusted odds ratios for all psychosis, non-affective psychosis, and affective psychosis. Maternal bereavement stress occurring preconception or during the prenatal period was not associated with a significant excess risk of psychosis in offspring (adjusted odds ratio, preconception 1.24, 95% confidence interval 0.96 to 1.62; first trimester 0.95, 0.58 to 1.56; second trimester 0.79, 0.46 to 1.33; third trimester 1.14, 0.78 to 1.66). Risks increased modestly after exposure to the loss of a close family member from birth to adolescence for all psychoses (adjusted odds ratio 1.17, 1.04 to 1.32). The pattern of risk was generally similar for non-affective and affective psychosis. Thus estimates were higher after death in the nuclear compared with extended family but remained non-significant for prenatal exposure; the earlier the exposure to death in the nuclear family occurred in childhood (all psychoses: adjusted odds ratio, birth to 2.9 years 1.84, 1.41 to 2.41; 3-6.9 years 1.47, 1.16 to 1.85; 7-12.9 years 1.32, 1.10 to 1.58) and after suicide. Following suicide, risks were especially higher for affective psychosis (birth to 2.9 years 3.33, 2.00 to 5.56; 6.9 years 1.84, 1.04 to 3.25; 7-12.9 years 2.68, 1.84 to 3.92). Adjustment for key confounders attenuated but did not explain associations with risk. Postnatal but not prenatal bereavement stress in mothers is associated with an increased risk of psychosis in offspring. Risks are especially high for affective psychosis after suicide in the nuclear family, an effect that is not explained by family psychiatric history. Future studies are needed to understand possible sources of risk and resilience so that structures can be put in place to support vulnerable children and their families.


Phase II trial of combretastatin A4 phosphate, carboplatin, and paclitaxel in patients with platinum-resistant ovarian cancer.
A previous dose-escalation trial of the vascular disrupting agent combretastatin A4 phosphate (CA4P) given before carboplatin, paclitaxel, or both showed responses in 7 of 18 patients with relapsed ovarian cancer. Patients with ovarian cancer that had relapsed and who could start trial therapy within 6 months of their last platinum chemotherapy were given CA4P 63 mg/m(2) minimum 18 h before paclitaxel 175 mg/m(2) and carboplatin AUC (area under the concentration curve) 5, repeated every 3 weeks. Five of the first 18 patients' disease responded, so the study was extended and closed after 44 patients were recruited. Grade ≥2 toxic effects were neutropenia in 75% and thrombocytopenia in 9% of patients (weekly blood counts), tumour pain, fatigue, and neuropathy, with one patient with rapidly reversible ataxia. Hypertension (23% of patients) was controlled by glyceryl trinitrate or prophylactic amlodipine. The response rate by RECIST was 13.5% and by Gynecologic Cancer InterGroup CA 125 criteria 34%. The addition of CA4P to paclitaxel and carboplatin is well tolerated and appears to produce a higher response rate in this patient population than if the chemotherapy was given without CA4P. A planned randomised trial will test this hypothesis.


Geochronology of cave deposits at Liang Bua and of adjacent river terraces in the Wae Racang valley, western Flores, Indonesia: a synthesis of age estimates for the type locality of Homo floresiensis.
A robust timeframe for the extant cave deposits at Liang Bua, and for the river terraces in the adjoining Wae Racang valley, is essential to constrain the period of existence and time of extinction of Homo floresiensis and other biota that have been excavated at this hominin type locality. Reliable age control is also required for the variety of artifacts excavated from these deposits, and to assist in environmental reconstructions for this river valley and for the region more broadly. In this paper, we summarize the available geochronological information for Liang Bua and its immediate environs, obtained using seven numerical-age methods: radiocarbon, thermoluminescence, optically- and infrared-stimulated luminescence (collectively known as optical dating), uranium-series, electron spin resonance, and coupled electron spin resonance/uranium-series. We synthesize the large number of numerical age determinations reported previously and present additional age estimates germane to questions of hominin evolution and extinction.


Surface plasmon-enhanced nanopillar photodetectors.
We demonstrate nanopillar-(NP) based plasmon-enhanced photodetectors (NP-PEPDs) operating in the near-infrared spectral regime. A novel fabrication technique produces subwavelength elongated nanoholes in a metal surface self-aligned to patterned NP arrays that acts as a 2D plasmonic crystal. Surface plasmon Polariton Bloch waves (SPP-BWs) are excited by the metal nanohole array resulting in electric field intensity "hot spots" in the NP. The NP periodicity determines the peak responsivity wavelength while the nanohole asymmetry produces polarization-dependent coupling of the SPP-BW modes. Resulting photodetectors have 0.28 A/W responsivity peaked at 1100 nm at a reverse bias of -5 V. Designs for further increasing the optical coupling efficiency into the nanopillar are explored. This technology has potential applications for plasmonically enhanced focal plane arrays and plasmonic photovoltaics.


Predicting metal-metal interactions. II. Accelerating generalized schemes through physical insights.
Operando-computational frameworks that integrate descriptors for catalyst stability within catalyst screening paradigms enable predictions of rates and selectivity on chemically faithful representations of nanoparticles under reaction conditions. These catalyst stability descriptors can be efficiently predicted by density functional theory (DFT)-based models. The alloy stability model, for example, predicts the stability of metal atoms in nanoparticles with site-by-site resolution. Herein, we use physical insights to present accelerated approaches of parameterizing this recently introduced alloy-stability model. These accelerated approaches meld quadratic functions for the energy of metal atoms in terms of the coordination number with linear correlations between model parameters and the cohesive energies of bulk metals. By interpolating across both the coordination number and chemical space, these accelerated approaches shrink the training set size for 12 fcc p- and d-block metals from 204 to as few as 24 DFT calculated total energies without sacrificing the accuracy of our model. We validate the accelerated approaches by predicting adsorption energies of metal atoms on extended surfaces and 147 atom cuboctahedral nanoparticles with mean absolute errors of 0.10 eV and 0.24 eV, respectively. This efficiency boost will enable a rapid and exhaustive exploration of the vast material space of transition metal alloys for catalytic applications.


Zn(2+) slows down Ca(V)3.3 gating kinetics: implications for thalamocortical activity.
We employed whole cell patch-clamp recordings to establish the effect of Zn(2+) on the gating the brain specific, T-type channel isoform Ca(V)3.3 expressed in HEK-293 cells. Zn(2+) (300 microM) modified the gating kinetics of this channel without influencing its steady-state properties. When inward Ca(2+) currents were elicited by step depolarizations at voltages above the threshold for channel opening, current inactivation was significantly slowed down while current activation was moderately affected. In addition, Zn(2+) slowed down channel deactivation but channel recovery from inactivation was only modestly changed. Zn(2+) also decreased whole cell Ca(2+) permeability to 45% of control values. In the presence of Zn(2+), Ca(2+) currents evoked by mock action potentials were more persistent than in its absence. Furthermore, computer simulation of action potential generation in thalamic reticular cells performed to model the gating effect of Zn(2+) on T-type channels (while leaving the kinetic parameters of voltage-gated Na(+) and K(+) unchanged) revealed that Zn(2+) increased the frequency and the duration of burst firing, which is known to depend on T-type channel activity. In line with this finding, we discovered that chelation of endogenous Zn(2+) decreased the frequency of occurrence of ictal-like epileptiform discharges in rat thalamocortical slices perfused with medium containing the convulsant 4-aminopyridine (50 microM). These data demonstrate that Zn(2+) modulates Ca(V)3.3 channel gating thus leading to increased neuronal excitability. We also propose that endogenous Zn(2+) may have a role in controlling thalamocortical oscillations.


Aerosol therapy by pressured metered-dose inhaler-spacer in sleeping young children: to do or not to do?
One third of young children are distressed during inhalation therapy. It has been suggested that administration during sleep could be a good alternative for these children. A laboratory study in our department using an infant upper airway model showed significantly higher lung doses from a pressured metered-dose inhaler (pMDI)-spacer for sleep-breathing patterns compared with wake-breathing patterns. We set up a daily life study to investigate the feasibility of aerosol administration by means of pMDI-spacer in sleeping young children. Over a period of 3 weeks, 30 children (age range, 6 to 23 months) with recurrent wheeze daily inhaled 1 puff of budesonide aerosol (200 mug) while awake and 1 puff during sleep. Filters positioned between the chamber and the facemask trapped the budesonide aerosol. Parents scored the child's asthma symptoms, degree of cooperation, and feasibility of administration on diary cards. In 69% of the sleep administrations, the children woke up, and in 75% of these cases the children were distressed. The mean filter dose (expressed as the percentage of the nominal dose) while awake was 47%, and during sleep it was 16% (p = 0.007). The median within-subject dose variability while awake was 50%, and during sleep it was 110% (p = 0.007). Aerosol administration by means of pMDI-spacer during sleep offers no advantage and is not a feasible treatment option in most young children.


Assessment of the national school lunch program in a subset of schools in San Juan, Puerto Rico: participants vs. non-participants.
Extensive evaluations of the national school lunch program (NSLP) have been carried out on the U.S. mainland. Puerto Rico, a commonwealth of the U.S. is a participant in this program, but has never been included in assessment studies. Herein, we present assessment information and compare results with comparable mainland studies. Multiple 24-hr recall questionnaires were administered to groups of participating (P) and non-participating (NP) children in the lunch program at 3 educational levels. Comparisons were made for children within the study as well as between comparable children in mainland studies for total intake of several macro- and micro-nutrients, contribution of the lunch to the total daily intake and adherence to U.S. Recommended Dietary Allowances (RDA's) or to Dietary Reference Intakes (DRI's) including acceptable macronutrient distribution ranges (AMDR's). Target intakes were met by P for % of the RDA of energy from protein, for all water soluble vitamins, iron, zinc and cholesterol. P did not achieve target intakes for total energy, energy from carbohydrates and fat nor for fat soluble vitamins, calcium, magnesium, phosphorus, potassium and fiber. Recommended levels were exceeded for sodium, total fat and saturated fat. Comparing P vs NP, the vast majority of both groups fell within AMDR recommendations for macronutrients but not all micronutrients. For the most part, our results parallel those obtained in the National sample however, results suggest that P in the lunch program in Puerto Rico have a healthier intake of several nutrients than NP students. The unique feature of this study is that it is the first assessment of the NSLP in a completely Hispanic population.


Novice nurses' perceptions of acute situations - A phenomenographic study.
Today's nurses face complex patient challenges and increased patient acuity. Novice nurses may feel unprepared for some of the tasks facing them. In order to prevent adverse events and improve patient outcomes, the management of acute situations has been identified as an area needing attention. Despite this, it is not yet clear what novice nurses themselves perceive as an acute situation. Therefore, the aim of this study is to describe novice nurses' perceptions of acute situations. The study has a qualitative, descriptive design with a phenomenographic approach. Semi-structured interviews were conducted with twelve novice nurses, with less than a year of working experience, about their perceptions of acute situations. Acute situations are perceived as situations that occur suddenly, that involve shortage of time, or that generate a sense of insufficient personal competence. When acute situations arise, nurses experience a feeling of overwhelming responsibility. Acute situations involve not only deteriorating patient health but can also include challenging interpersonal relations or deficiencies in organisational procedures. An acute situation, as perceived by novice nurses, contains significantly more than medical manifestations related to the patient, a prominent perception being that interpersonal relations are crucial for appropriate management.


Vascular anomalies and lymphedema.
After studying this article, the participant should be able to: 1. Define the difference between vascular tumors and malformations. 2. Distinguish between the natural history of hemangiomas and that of vascular malformations. 3. Identify the different types of hemangiomas and vascular malformations and understand evaluation, treatment, and complications. 4. Understand the role of lymphaticovenular anastomoses in the treatment of extremity lymphedema. The International Society for the Study of Vascular Anomalies classification, which is the most widely accepted classification system in use, divides vascular anomalies into vascular tumors (inclusive of hemangiomas) and malformations. This serves as a guideline for diagnosis, evaluation, and treatment of these lesions. Although hemangiomas tend to have a predictable clinical course over the first year of life, going through proliferating, involuting, and involuted stages, vascular malformations demonstrate growth commensurate with age, often becoming more prominent in puberty. In addition, they never regress, and persist throughout life. Different modalities of treatment may be appropriate for vascular tumors and different subsets of vascular malformations. Details are provided in this review. Lymphaticovenular anastomoses provide an excellent addition to our methods of treatment of extremity lymphedema, and are made possible through development of supermicrosurgical techniques. Vascular anomalies have a high prevalence in the general population. Thus, it is vital that the plastic surgeon has a good understanding of classification, evaluation, and treatment options. Lymphedema is another common condition that is encountered. Understanding of lymphaticovenular anastomoses and their applications aids treatment planning for select patients.


Phylogeographic structure and ecological niche modelling reveal signals of isolation and postglacial colonisation in the European stag beetle.
Lucanus cervus (L.), the stag beetle, is a saproxylic beetle species distributed widely across Europe. Throughout its distribution the species has exhibited pronounced declines and is widely considered threatened. Conservation efforts may be hindered by the lack of population genetic data and understanding of the spatial scale of population connectivity. To address this knowledge gap this research details the first broad scale phylogeographic study of L. cervus based on mitochondrial DNA (mtDNA) sequencing and microsatellite analysis of samples collected from 121 localities across Europe. Genetic data were complemented by palaeo-distribution models of spatial occupancy during the Last Glacial Maximum to strengthen inferences of refugial areas. A salient feature of the mtDNA was the identification of two lineages. Lineage I was widespread across Europe while lineage II was confined to Greece. Microsatellites supported the differentiation of the Greek samples and alongside palaeo-distribution models indicated this area was a glacial refuge. The genetic endemism of the Greek samples, and demographic results compatible with no signatures of spatial expansion likely reflects restricted dispersal into and out of the area. Lineage I exhibited a shallow star like phylogeny compatible with rapid population expansion across Europe. Demographic analysis indicated such expansions occurred after the Last Glacial Maximum. Nuclear diversity and hindcast species distribution models indicated a central Italian refuge for lineage I. Palaeo-distribution modelling results also suggested a western refuge in northern Iberia and south-west France. In conclusion the results provide evidence of glacial divergence in stag beetle while also suggesting high, at least on evolutionary timescales, gene flow across most of Europe. The data also provide a neutral genetic framework against which patterns of phenotypic variation may be assessed.


Dynein isoforms in sea urchin eggs.
Biochemical and immunological analysis of unfertilized sea urchin eggs has revealed the presence of at least two distinct isoforms of cytoplasmic dyneins, one soluble and the other microtubule-associated. The soluble enzyme is a 20 S particle with a MgATPase activity that can be activated 5-fold by nonionic detergents. It contains heavy chain polypeptides that 1) comigrate with the dynein heavy chains of embryonic cilia; 2) cross-react with antibodies against flagellar dynein; and 3) are cleaved by UV irradiation in the presence of MgATP and sodium vanadate into specific peptide fragments. The soluble egg dynein is, therefore, closely related to axonemal dynein and may be a ciliary precursor. Egg microtubule preparations contain a distinct dynein-like polypeptide, previously designated HMr-3 (Scholey, J.M., Neighbors, B., McIntosh, J.R., and Salmon, E.D. (1984) J. Biol Chem. 259, 6516-6525). HMr-3 binds microtubules in an ATP-sensitive fashion; it sediments at 20 S on sucrose density gradients, and it is susceptible to vanadate-sensitized UV cleavage. However, HMr-3 can be distinguished from the soluble cytoplasmic dynein on the basis of its weak cross-reactivity with antiflagellar dynein antibodies, its heavy chain composition on high resolution sodium dodecyl sulfate-polyacrylamide gel electrophoresis, its low specific ATPase activity, and the molecular weight of its vanadate-induced UV cleavage fragments. HMr-3 may represent a dynein-like polypeptide that is distinct from the pool of ciliary dynein precursors.


Long-term effects of prenatal stress on dopamine and glutamate receptors in adult rat brain.
Prenatal stress greatly influences the ability of an individual to manage stressful events in adulthood. Such vulnerability may result from abnormalities in the development and integration of forebrain dopaminergic and glutamatergic projections during the prenatal period. In this study, we assessed the effects of prenatal stress on the expression of selective dopamine and glutamate receptor subtypes in the adult offsprings of rats subjected to repeated restraint stress during the last week of pregnancy. Dopamine D2-like receptors increased in dorsal frontal cortex (DFC), medial prefrontal cortex (MPC), hippocampal CA1 region and core region of nucleus accumbens (NAc) of prenatally stressed rats compared to control subjects. Glutamate NMDA receptors increased in MPC, DFC, hippocampal CA1, medial caudate-putamen, as well as in shell and core regions of NAc. Group III metabotropic glutamate receptors increased in MPC and DFC of prenatally stressed rats, but remained unchanged in all other regions examined. These results indicate that stress suffered during the gestational period has long lasting effects that extend into the adulthood of prenatally stressed offsprings. Changes in dopamine and glutamate receptor subtype levels in different forebrain regions of adult rats suggest that the development and formation of the corticostriatal and corticolimbic pathways may be permanently altered as a result of stress suffered prenatally. Maldevelopment of these pathways may provide a neurobiological substrate for the development of schizophrenia and other idiopathic psychotic disorders.


Perceptions and Practices of Community Pharmacists towards Antimicrobial Stewardship in the State of Selangor, Malaysia.
Increasing antimicrobial resistance is one of the pressing concerns globally. Injudicious use of antibiotics is one of the modifiable factors responsible for antimicrobial resistance. Given the widespread use of antimicrobials in community settings, pharmacists have an important role in ensuring appropriate use of antibiotics. The objective of this study was to assess the perception and self-reported practices of community pharmacists towards antimicrobial stewardship. A cross-sectional study was conducted among community pharmacists between March-April, 2015, using a self-administered, pre-tested questionnaire in the State of Selangor, Malaysia. A simple random sampling approach was used to select pharmacy sites. Descriptive and inferential statistical methods were used to analyse the data. A total of 188 pharmacists responded to the survey, giving a response rate of 83.5%. The majority of participants (n = 182, 96.8%) believed that antimicrobial stewardship program helps healthcare professionals to improve the quality of patient care. However, more than half of pharmacists were neutral in their opinion about the incorporation of antimicrobial stewardship programs in community pharmacies (n = 102, 54.2%). Though collaboration was often done by pharmacists with other health professionals over the use of antibiotics (n = 104, 55.3%), a significant proportion of participants (n = 102, 54.2%) rarely/occasionally participate in antimicrobial awareness campaigns. Pharmacists having postgraduate qualification were more likely to held positive perceptions of, and were engaged in, antimicrobial stewardship than their non-postgraduate counterpart (p<0.05). Similarly, more experienced pharmacists (> 10 years) held positive perceptions towards antimicrobial stewardship (p<0.05). The study highlighted some gaps in the perception and practices of community pharmacist towards antimicrobial stewardship. Development of customized interventions would be critical to bridging these gaps and improve their perception and practices towards antimicrobial stewardship.


The Benefits of Benevolence: Basic Psychological Needs, Beneficence, and the Enhancement of Well-Being.
Pro-social behaviors have been associated with enhanced well-being, but what psychological mechanisms explain this connection? Some theories suggest that beneficence-the sense of being able to give-inherently improves well-being, whereas evidence from self-determination theory (Weinstein & Ryan, 2010) shows that increases in well-being are mediated by satisfaction of innate psychological needs for autonomy, competence, and relatedness. Here we simultaneously assess these two explanations. Study 1 (N = 335) used a cross-sectional survey with an Internet sample to develop a measure to assess beneficence satisfaction. The next two cross-sectional Internet-sample studies tested mediators between pro-social behavior and general well-being (Study 2, N = 332) and situational peak moment well-being (Study 3, N = 180). A fourth study (N = 85) used a diary method with university students to assess daily fluctuations in well-being associated with needs and beneficence. It was shown across all studies that both the three psychological needs and beneficence satisfaction mediate the relations between pro-social actions and well-being, with all four factors emerging as independent predictors. Together, these studies underscore the role of autonomy, competence, and relatedness in explaining the well-being benefits of benevolence, and they also point to the independent role of beneficence as a source of human wellness.


The epidemiology and health care burden of tennis elbow: a population-based study.
Lateral elbow tendinosis (epicondylitis) is a common condition both in primary care and specialty clinics. To evaluate the natural history (ie, incidence, recurrence, and progression to surgery) of lateral elbow tendinosis in a large population. Descriptive epidemiology study. The study population comprised a population-based incidence cohort of patients with new-onset lateral elbow tendinosis between January 1, 2000, and December 31, 2012. The medical records of a 10% random sample (n=576) were reviewed to ascertain information on patient and disease characteristics, treatment modalities, recurrence, and progression to surgery. Age- and sex-specific incidence rates were calculated and adjusted to the 2010 US population. The age- and sex-adjusted annual incidence of lateral elbow tendinosis decreased significantly over time from 4.5 per 1000 people in 2000 to 2.4 per 1000 in 2012 (P<.001). The recurrence rate within 2 years was 8.5% and remained constant over time. The proportion of surgically treated cases within 2 years of diagnosis tripled over time, from 1.1% during the 2000-2002 time period to 3.2% after 2009 (P<.00001). About 1 in 10 patients with persistent symptoms at 6 months required surgery. The decrease in incidence of lateral elbow tendinosis may represent changes in diagnosis patterns or a true decrease in disease incidence. Natural history data can be used to help guide patients and providers in determining the most appropriate course at a given time in the disease process. The study data suggest that patients without resolution after 6 months of onset may have a prolonged disease course and may need surgical intervention.


Scintigraphic and radiographic evaluation of appendicular skeletal lesions in cold-stunned Kemp's ridley sea turtles.
Osteolytic appendicular skeletal lesions in eight-stranded, cold-stunned Kemp's ridley sea turtles (Lepidochelys kempii) were evaluated using radiography and skeletal scintigraphy. Radiographic studies were performed monthly in most animals. Follow-up scintigraphy was performed 45-120 days after the initial exams in six turtles. Radiographically, lesions slowly progressed from an early osteolytic process contained to either the proximal or distal end of long bones, to a later stage characterized by thickening of the affected bone, sclerosis, and remodeling of the lesion borders. In seven turtles, the initial scintigrams were characterized by at least one focus of abnormal radiopharmaceutical uptake that correlated with a lytic site noted in radiographs. In five turtles, scintigraphic lesions were characterized by asymmetric radiopharmaceutical uptake rather than by increased intensity of uptake. Scintigraphic studies obtained more than 4 months after the appearance of clinical and radiographic signs had minimal, if any, abnormal radiopharmaceutical uptake, despite the persistence of abnormal radiographic findings. Skeletal scintigraphy is an effective method for more precisely determining if and when these animals can be returned to the wild. Animals were released if normal radiopharmaceutical uptake was seen during initial examination, or if decreased uptake was noted between serial examinations. In four of the turtles, resolution of abnormal scintigraphic findings permitted an objective decision to discontinue antibiotic and antifungal therapy. Seven of the eight turtles were released after correlation of the clinical signs with the imaging findings. Radiographs, however, are still needed to facilitate the correct identification of lesions with scintigraphy.


COVID-19 Vaccines: Preparing for Vaccination in the Context of Clinical Oncology Care.
In the environment of an infectious pandemic, vaccines are a primary public health strategy to prevent the spread of disease. With the COVID-19 pandemic, there is heightened interest in safe and effective vaccines and their use in the context of clinical oncology practice. This article provides foundational information about vaccines in general and vaccines developed to protect against the SARS-CoV-2 virus in the United States, as well as clinical nurse strategies to apply vaccines in clinical oncology practice. The article is based on a review of public health literature and reputable websites about vaccines and their development in clinical care. This foundational information about vaccines reviews their history and development, as well as the development of COVID-19 vaccines specifically, and discusses COVID-19 vaccines as part of clinical oncology care. Supporting best practices in clinical oncology care, nurses can provide factual, evidence-based information about vaccine safety, effectiveness, and safe administration.


Research Techniques Made Simple: Use of Imaging Mass Cytometry for Dermatological Research and Clinical Applications.
Traditional immunohistochemistry (IHC) is inherently limited by its ability to analyze only several markers within a histological tissue section at a given time, which hinders in-depth characterization and phenotyping of tissues. Imaging mass cytometry (IMC), which combines IHC using metal-labeled antibodies with laser ablation and detection using mass cytometry by time-of-flight, overcomes this limitation with the capability to simultaneously analyze up to 40 protein markers to generate high-dimensional images from a single tissue section. IMC analysis preserves tissue architecture and spatial cellular relationships that would otherwise be lost or significantly altered in applications requiring tissue dissociation, such as flow cytometry or single-cell RNA sequencing. Resulting high-dimensional histological images permit spatially conserved analysis to identify unique cell populations, cellular interactions and avoidances, and insight into activation and behavioral status based on tissue location. IMC can be performed on both frozen and formalin-fixed paraffin-embedded tissue, allowing for previously banked samples to be analyzed and correlated with known clinical outcomes. Expectedly, IMC will change the landscape of investigative pathology, particularly when used in coordination with multiomic platforms to combine transcriptomic and proteomic data at a single-cell resolution. Here, we aim to highlight the potential utility of IMC within dermatologic research and clinical applications.


Increased DNA damage sensitivity and apoptosis in cells lacking the Snf5/Ini1 subunit of the SWI/SNF chromatin remodeling complex.
The gene encoding the SNF5/Ini1 core subunit of the SWI/SNF chromatin remodeling complex is a tumor suppressor in humans and mice, with an essential role in early embryonic development. To investigate further the function of this gene, we have generated a Cre/lox-conditional mouse line. We demonstrate that Snf5 deletion in primary fibroblasts impairs cell proliferation and survival without the expected derepression of most retinoblastoma protein-controlled, E2F-responsive genes. Furthermore, Snf5-deficient cells are hypersensitive to genotoxic stress, display increased aberrant mitotic features, and accumulate phosphorylated p53, leading to elevated expression of a specific subset of p53 target genes, suggesting a role for Snf5 in the DNA damage response. p53 inactivation does not rescue the proliferation defect caused by Snf5 deficiency but reduces apoptosis and strongly accelerates tumor formation in Snf5-heterozygous mice.


Steady state anisotropy two-photon microscopy resolves multiple, spectrally similar fluorophores, enabling in vivo multilabel imaging.
The use of spectrally distinguishable fluorescent dyes enables imaging of multiple targets. However, in two-photon microscopy, the number of fluorescent labels with distinct emission spectra that can be effectively excited and resolved is constrained by the confined tuning range of the excitation laser and the broad and overlapping nature of fluorophore two-photon absorption spectra. This limitation effectively reduces the number of available imaging channels. Here, we demonstrate that two-photon steady state anisotropy imaging (2PSSA) offers the capability to resolve otherwise unresolvable fluorescent tracers both in live cells and in mouse tumor models. This approach expands the number of biological targets that can be imaged simultaneously, increasing the total amount of information that can be obtained through imaging.


Evaluation of antipyretic potential of Nelumbo nucifera stalk extract.
The ethanol extract of stalks of Nelumbo nucifera (NNSE) was evaluated for its antipyretic potential on normal body temperature and yeast induced pyrexia in rats. NNSE showed significant activity in both the models at oral doses of 200 and 400 mg/kg. NNSE at a dose of 200 mg/kg was found to produce significant lowering of normal body temperature up to 3 h and at 400 mg/kg it caused significant lowering of body temperature up to 6 h after its administration. In the model of yeast provoked elevation of body temperature NNSE showed dose-dependent lowering of body temperature up to 4 h at both the doses and the results were comparable to that of paracetamol, a standard antipyretic agent.


[Central nervous system involvement of sarcoidosis: diagnosis and management].
Sarcoidosis is a multisystem inflammatory granulomatous disease that affects the central nervous system (CNS). CNS Involvement occurs in a relatively small number of patients with sarcoidosis but is potentially serious manifestation. Diagnostic criteria usually include histologic identification of a noncaseating epithelioid granuloma, supportive laboratory or neuroimaging tests or both, and a compatible clinical course. Diagnostic criteria proposed by "The Diagnosis Guideline for Sarcoidosis in Japan -2006" can distinguish definite, probable, and possible neurosarcoidosis, and may be now commonly used. Patients with a definite or probable diagnosis of CNS sarcoidosis should start treatment promptly. The pharmacologic regimens for the treatment of neurosarcoidosis are not standardized since no prospective, randomized, controlled trials have been performed in the World. Corticosteroids, however, are typically the first line or therapy and approximately half of patients have substantial benefit. For patients who are refractory to or intolerant of corticosteroid therapy, second-line agents include azathioprine, methotrexate, cyclosporine, cyclophosphamide, and mycophenolate mofetil (MMF). Treatment regimens vary in term of doing, maintenance, and tapering. Radiotherapy or neurosurgical treatment is indicated when medications fail or when life-threatening emergences, such as severe hydrocephalus and elevated intracranial pressure, arise. The combination of infliximab, an anti-TNF-alpha neutralizing antibody, and MMF has recently been used to treat neurosarcoidosis. Treatment option will likely evolve as well-designed studies are undertaken.


3D simulation of the image formation in soft x-ray microscopes.
In water-window soft x-ray microscopy the studied object is typically larger than the depth of focus and the sample illumination is often partially coherent. This blurs out-of-focus features and may introduce considerable fringing. Understanding the influence of these phenomena on the image formation is therefore important when interpreting experimental data. Here we present a wave-propagation model operating in 3D for simulating the image formation of thick objects in partially coherent soft x-ray microscopes. The model is compared with present simulation methods as well as with experiments. The results show that our model predicts the image formation of transmission soft x-ray microscopes more accurately than previous models.


The neuropsychology of first impressions: Evidence from Huntington's disease.
Impairments of emotion recognition have been widely documented in Huntington's disease (HD), but little is known concerning how these relate to other aspects of social cognition, including first impressions of traits such as trustworthiness and dominance. Here, we introduce a novel and sensitive method to investigate the ability to evaluate trustworthiness and dominance from facial appearance, with control tasks measuring ability to perceive differences between comparable stimuli. We used this new method together with standard tests of face perception to investigate social cognition in HD. We found that a subgroup of people with HD was impaired at perceiving trustworthiness and dominance, and that perceiving trustworthiness and dominance were correlated with impaired facial expression recognition. In addition, we used diffusion tensor imaging (DTI) to provisionally identify candidate brain regions associated with social cognition by contrasting regional functional anisotropy (FA) measures between subgroups of HD participants showing normal or impaired perception of trustworthiness and dominance, and by correlating these regional brain abnormalities with behavioural performance on tests of emotion recognition. In this way we show for the first time alterations in perception of trustworthiness and dominance in people with HD and link these to regions which may map the boundaries of the social brain. The pattern of breakdown seen in this neurodegenerative disease can thus be used to explore potential inter-relationships between different components of social cognition.


Short-term weight gain after antiretroviral therapy initiation and subsequent risk of cardiovascular disease and diabetes: the D:A:D study.
The aim of the study was to assess the impact of the gain in body mass index (BMI) observed immediately after antiretroviral therapy (ART) initiation on the subsequent risk of cardiovascular disease (CVD) and diabetes. We analysed data from the Data Collection on Adverse Events of Anti-HIV Drugs (D:A:D) cohort study. Outcomes were development of (i) CVD (composite of myocardial infarction/stroke/coronary procedure) and (ii) diabetes. The main exposure variable was change in BMI from ART initiation (pre-ART) to 1 year after initiation (continuous variable) in treatment-naïve individuals initiating ART with no history of CVD or diabetes (for respective outcomes). BMI [weight (kg)/(height (m))(2)] was categorized as underweight (< 18.5), normal (18.5-25), overweight (25-30) and obese (> 30). Poisson regression models were fitted stratified for each pre-ART BMI category to allow for category-specific estimates of incidence rate ratio (IRR). Models were adjusted for pre-ART BMI and CD4 count, key known risk factors (time-updated where possible) and calendar year. A total of 97 CVD events occurred in 43,982 person-years (n = 9321) and 125 diabetes events in 43,278 person-years (n = 9193). In fully adjusted analyses for CVD, the IRR/unit gain in BMI (95% confidence interval) in the first year of ART, by pre-ART BMI category, was: underweight, 0.90 (0.60-1.37); normal, 1.18 (1.05-1.33); overweight, 0.87 (0.70-1.10), and obese, 0.95 (0.71-1.28) (P for interaction = 0.04). For diabetes, the IRR/unit gain in BMI was 1.11 (95% confidence interval 1.03 to 1.21), regardless of pre-ART BMI (P for interaction > 0.05). Short-term gain in BMI following ART initiation appeared to increase the longer term risk of CVD, but only in those with pre-ART BMI in the normal range. It was also associated with increased risk of diabetes regardless of pre-ART BMI.


Physiological basis for an animal model of the renal Fanconi syndrome: use of succinylacetone in the rat.
1. The biochemical basis for the human renal Fanconi syndrome, including glucosuria, phosphaturia and aminoaciduria, remains enigmatic. This is due, in part, to the lack of an appropriate animal model. Since there is an association between the human genetic disease hereditary tyrosinaemia, for which urinary excretion of the compound succinylacetone constitutes a biochemical marker, and a renal Fanconi syndrome, we have examined the relationship between succinylacetone and renal tubular function in the rat. 2. Intraperitoneal injection of succinylacetone for 3 consecutive days into adult male Sprague-Dawley rats resulted in succinylacetone plasma concentration of 3 mmol/l. This concentration was associated with glucosuria, aminoaciduria, polyuria, reduced renal phosphate reabsorption and normal creatinine clearance. In addition, urinary porphobilinogen and total porphyrin excretions were markedly reduced. In animals permitted to recover for 7 days after succinylacetone administration, these renal functional changes remitted partially or completely. Ultrastructural examination of the kidneys after the 3 days of treatment showed no fine structural changes. 3. We conclude that the physiological alterations produced in normal rat renal tubules by succinylacetone provide the basis for the study of the biochemical changes underlying the human renal Fanconi syndrome.


Optimizing Modern Family Size: Trade-offs between Fertility and the Economic Costs of Reproduction.
Modern industrialized populations lack the strong positive correlations between wealth and reproductive success that characterize most traditional societies. While modernization has brought about substantial increases in personal wealth, fertility in many developed countries has plummeted to the lowest levels in recorded human history. These phenomena contradict evolutionary and economic models of the family that assume increasing wealth reduces resource competition between offspring, favoring high fertility norms. Here, we review the hypothesis that cultural modernization may in fact establish unusually intense reproductive trade-offs in wealthy relative to impoverished strata, favoring low fertility. We test this premise with British longitudinal data (the Avon Longitudinal Study of Parents and Children), exploring maternal self-perceptions of economic hardship in relation to increasing family size and actual socioeconomic status. Low-income and low-education-level mothers perceived the greatest economic costs associated with raising two versus one offspring. However, for all further increases to family size, reproduction appears most expensive for relatively wealthy and well-educated mothers. We discuss our results and review current literature on the long-term consequences of resource dilution in modern families.


An Analysis of the Massachusetts Healthcare Law.
Healthcare in the United States has been one topic of the debates and discussion in the country for many years. The challenge for affordable, accessible, and quality healthcare for most Americans has been on the agenda of federal and state legislatures. There is probably no other state that has drawn as much individual attention regarding this challenge as the state of Massachusetts. While researching the topic for this article, it was discovered that financial and political perspectives on the success or failure of the healthcare model in Massachusetts vary depending on the aspect of the system being discussed. In this article the authors give a brief history and description of the Massachusetts Healthcare Law, explanation of how the law is financed, identification of the targeted populations in Massachusetts for which the law provides coverage, demonstration of the actual benefit coverage provided by the law, and review of the impact of the law on healthcare providers such as physicians and hospitals. In addition, there are explanations about the impact of the law on health insurance companies, discussion of changes in healthcare premiums, explanation of costs to the state for the new program, reviews of the impact on the health of the insured, and finally, projections on the changes that healthcare facilities will need to make to maintain fiscal viability as a result of this program.


Topical fluoroquinolones for eye and ear.
Topical fluoroquinolones are now available for use in the eye and ear. Their broad spectrum of activity includes the common eye and ear pathogens Staphylococcus aureus and Pseudomonas aeruginosa. For the treatment of acute otitis externa, these agents are as effective as previously available otic preparations. For the treatment of otitis media with tympanic membrane perforation, topical fluoroquinolones are effective and safe. These preparations are approved for use in children, and lack of ototoxicity permits prolonged administration when necessary. Topical fluoroquinolones are not appropriate for the treatment of uncomplicated conjunctivitis where narrower spectrum agents suffice; they represent a simplified regimen for the treatment of bacterial keratitis (corneal ulcers). When administered topically, fluoroquinolones are well tolerated and offer convenient dosing schedules. Currently, bacterial resistance appears limited.


Immunoblot analyses of Candida albicans-associated antigens and antibodies in human sera.
We tested 10 patient sera for the presence of immunoglobulin G (IgG) antibodies to Candida albicans and for C. albicans antigens by immunoblot analysis (i.e., electrotransfer blot radioimmunoassay) (G. E. Smith and M. D. Summers, J. Virol. 39:125-137, 1981). We evaluated sera from two patients at risk for candidiasis, five patients with systemic candidiasis documented by culture, and two patients who had experienced transient candidemia. Both the specificity and the relative amount of IgG antibodies to C. albicans in each serum sample were readily visualized by this technique, as was the absence of antibody from serum of neonatal and immunocompromised patients. No antibody species appeared to be uniquely associated with candidiasis patients (i.e., each antibody species present in the candidiasis patient was also present in sera of normal individuals or "at-risk" patients). IgG from rabbits immunized with whole cells or with a cytoplasmic fraction of C. albicans was used to detect C. albicans antigens in patient sera. Although several antigens were detected in the sera from patients with candidiasis, the same antigens were also detected in sera from patients at risk and in normal human serum. No antigens were detected in human serum when preimmune rabbit sera were used. These results suggest that the antigens detected by the rabbit antisera were human serum proteins that cross-reacted with C. albicans antigens. These findings may have important implications in studies of both the pathobiology of C. albicans and the serodiagnosis of candidiasis.


Genomic sequence correction by single-stranded DNA oligonucleotides: role of DNA synthesis and chemical modifications of the oligonucleotide ends.
Single-stranded oligonucleotides (ssODN) can induce site-specific genetic alterations in selected mammalian cells, but the involved mechanisms are not known. We corroborate the potential of genomic sequence correction by ssODN using chromosomally integrated mutated enhanced green fluorescent protein (mEGFP) reporter genes in CHO cell lines. The role of integration site was studied in a panel of cell clones with randomly integrated reporters and in cell lines with site-specific single copy integration of the mEGFP reporter in opposite orientations. Involvement of end modification was examined on ssODN with unprotected or phosphorothioate (PS) protected ends. Also ssODN containing octyl or hexaethylene glycol (HEG) end blocking groups were tested. The significance of DNA synthesis was investigated by cell cycle analysis and by the DNA polymerases alpha, delta and epsilon inhibitor aphidicolin. Correction rates of up to 5% were observed upon a single transfection of ssODN. Independent of the mEGFP chromosomal integration site and of its orientation towards the replication fork, antisense ssODN were more effective than sense ssODN. When ssODN ends were blocked by either octyl or HEG groups, correction rates were reduced. Finally, we demonstrate a dependence of the process on DNA synthesis. We show that, on a chromosomal level, the orientation of the replication fork towards the targeted locus is not central in the strand bias of ssODN-based targeted sequence correction. We demonstrate the importance of accessible ssODN ends for sequence alteration. Finally, we provide evidence for the involvement of DNA synthesis in the process.


Evolutionary determinants of cancer.
Our understanding of cancer is being transformed by exploring clonal diversity, drug resistance, and causation within an evolutionary framework. The therapeutic resilience of advanced cancer is a consequence of its character as a complex, dynamic, and adaptive ecosystem engendering robustness, underpinned by genetic diversity and epigenetic plasticity. The risk of mutation-driven escape by self-renewing cells is intrinsic to multicellularity but is countered by multiple restraints, facilitating increasing complexity and longevity of species. But our own species has disrupted this historical narrative by rapidly escalating intrinsic risk. Evolutionary principles illuminate these challenges and provide new avenues to explore for more effective control. Lifetime risk of cancer now approximates to 50% in Western societies. And, despite many advances, the outcome for patients with disseminated disease remains poor, with drug resistance the norm. An evolutionary perspective may provide a clearer understanding of how cancer clones develop robustness and why, for us as a species, risk is now off the scale. And, perhaps, of what we might best do to achieve more effective control.


A provisional regulatory gene network for specification of endomesoderm in the sea urchin embryo.
We present the current form of a provisional DNA sequence-based regulatory gene network that explains in outline how endomesodermal specification in the sea urchin embryo is controlled. The model of the network is in a continuous process of revision and growth as new genes are added and new experimental results become available; see http://www.its.caltech.edu/~mirsky/endomeso.htm (End-mes Gene Network Update) for the latest version. The network contains over 40 genes at present, many newly uncovered in the course of this work, and most encoding DNA-binding transcriptional regulatory factors. The architecture of the network was approached initially by construction of a logic model that integrated the extensive experimental evidence now available on endomesoderm specification. The internal linkages between genes in the network have been determined functionally, by measurement of the effects of regulatory perturbations on the expression of all relevant genes in the network. Five kinds of perturbation have been applied: (1) use of morpholino antisense oligonucleotides targeted to many of the key regulatory genes in the network; (2) transformation of other regulatory factors into dominant repressors by construction of Engrailed repressor domain fusions; (3) ectopic expression of given regulatory factors, from genetic expression constructs and from injected mRNAs; (4) blockade of the beta-catenin/Tcf pathway by introduction of mRNA encoding the intracellular domain of cadherin; and (5) blockade of the Notch signaling pathway by introduction of mRNA encoding the extracellular domain of the Notch receptor. The network model predicts the cis-regulatory inputs that link each gene into the network. Therefore, its architecture is testable by cis-regulatory analysis. Strongylocentrotus purpuratus and Lytechinus variegatus genomic BAC recombinants that include a large number of the genes in the network have been sequenced and annotated. Tests of the cis-regulatory predictions of the model are greatly facilitated by interspecific computational sequence comparison, which affords a rapid identification of likely cis-regulatory elements in advance of experimental analysis. The network specifies genomically encoded regulatory processes between early cleavage and gastrula stages. These control the specification of the micromere lineage and of the initial veg(2) endomesodermal domain; the blastula-stage separation of the central veg(2) mesodermal domain (i.e., the secondary mesenchyme progenitor field) from the peripheral veg(2) endodermal domain; the stabilization of specification state within these domains; and activation of some downstream differentiation genes. Each of the temporal-spatial phases of specification is represented in a subelement of the network model, that treats regulatory events within the relevant embryonic nuclei at particular stages.


Patient Experience with Conscious Sedation as a Method of Pain Relief for Transvaginal Oocyte Retrieval: A Cross Sectional Study.
The aim of the study was to measure patient's satisfaction level and acceptance of conscious sedation as a method of pain relief following transvaginal oocyte retrieval (TVOR) during assisted reproduction technology treatment. We also evaluated the factors that may influence the efficacy of conscious sedation method. A prospective cross-sectional study. Prospective study was conducted from October 2015 to January 2016 at a university-level hospital and 100 women were recruited. Variables for analysis included woman age, duration of procedure, number of oocytes retrieved, and transmyometrial passage of the needle. Pain assessment was done by visual analog scale (VAS). Medical complications, and patient satisfaction score [Likert's score and client satisfaction questionnaire (CSQ)] were recorded. There was a moderate positive correlation between age and pain score on day 1 post-procedure. When the duration of procedure was >12 min, immediate post-procedure pain score was significantly higher compared to those whose procedure where duration was <12 min. There was no correlation between pain score and the number of oocytes retrieved (≤5, 6-15, and ≥16) and transmyometrial passage of needle. The VAS 10-point score immediately post-procedure, after 6 and 24 h post-procedure, and on day of embryo transfer was 2.83 (±1.67), 0.78 (±1.04), 0.39 (±1.09), and 0.14 (±0.58), respectively. The Likert's score was 3.65 (±0.82) and mean CSQ was 27.04 (±3.01). Majority of the women (86%) preferred the same pain relief method for future analgesia. There were no major complications. Conscious sedation was associated with high satisfaction level and acceptance rate among patients undergoing TVOR.


Combined Peptidomic and Proteomic Analysis of Electrically Stimulated and Manually Dissected Venom from the South American Bullet Ant Paraponera clavata.
Ants have evolved venoms rich in peptides and proteins used for predation, defense, and communication. However, they remain extremely understudied due to the minimal amount of venom secreted by each ant. The present study investigated the differences in the proteome and peptidome of the venom from the bullet ant, Paraponera clavata. Venom samples were collected from a single colony either by manual venom gland dissection or by electrical stimulation and were compared using proteomic methods. Venom proteins were separated by 2D-PAGE and identified by nanoLC-ESI-QTOF MS/MS. Venom peptides were initially separated using C18 reversed-phase high-performance liquid chromatography, then analyzed by MALDI-TOF MS. The proteomic analysis revealed numerous proteins that could be assigned a biological function (total 94), mainly as toxins, or roles in cell regulation and transport. This investigation found that ca. 73% of the proteins were common to venoms collected by the two methods. The peptidomic analysis revealed a large number of peptides (total 309) but with <20% shared by the two collection methods. There was also a marked difference between venoms obtained by venom gland dissection from different ant colonies. These findings demonstrate the rich composition and variability of P. clavata venom.


The human SRY protein is present in fetal and adult Sertoli cells and germ cells.
Sex determination in mammals is controlled by the Y chromosome located SRY gene. Despite recent advances towards understanding the mechanisms that regulate sex determination in mammals, the expression profile of the SRY protein in human tissues is unknown. To localize the SRY protein and determine its cellular distribution, we prepared monoclonal antibodies (mAb) against the recombinant SRY protein. One antibody, LSRY1.1, recognizes a SRY-specific epitope and was used to localize the protein in different cells and tissues. The mAb recognizes a protein of 27 kDa in total lysates of HeLa SRYB3 cells. Immunocytochemical staining showed a nuclear localization of the protein. Immunohistochemical studies performed on gonadal tissue of a fetus, a one month-old boy and an adult man, demonstrated the presence of SRY protein in the nucleus of Sertoli and germ cells. In addition two 46,XX SRY(+) males had the SRY protein in their gonadal tissues. All other samples were negative, including all female tissue studied and the testis of a 46,XX SRY(-) male. The presence of SRY protein in fetal and adult gonadal tissues including germ cells suggests that SRY may have other male-specific functions in addition to sex determinism.


Effects of peripheral sensory nerve stimulation plus task-oriented training on upper extremity function in patients with subacute stroke: a pilot randomized crossover trial.
To investigate the feasibility of peripheral sensory nerve stimulation combined with task-oriented training in patients with stroke during inpatient rehabilitation. A pilot randomized crossover trial. Two rehabilitation hospitals. Twenty-two patients with subacute stroke. Participants were randomly assigned to two groups and underwent two weeks of training in addition to conventional inpatient rehabilitation. The immediate group underwent peripheral sensory nerve stimulation combined with task-oriented training in the first week, followed by another week with task-oriented training alone. The delayed group underwent the same training in reverse order. Outcome measures were the level of fatigue and Wolf Motor Function Test. Patients were assessed at baseline, one and two weeks. All participants completed the study with no adverse events. There was no significant difference in level of fatigue between each treatment. From baseline to one week, the immediate group showed larger improvements than the delayed groups in the Wolf Motor Function Test (decrease in mean time (± SD) from 41.9 ± 16.2 seconds to 30.6 ± 11.4 seconds versus from 46.8 ± 19.4 seconds to 42.9 ± 14.7 seconds, respectively) but the difference did not reach significance after Bonferroni correction (P = 0.041). Within-group comparison showed significant improvements in the Wolf Motor Function Test mean time after the peripheral sensory nerve stimulation combined with task-oriented training periods in each group (P < 0.01). Peripheral sensory nerve stimulation is feasible in clinical settings and may enhance the effects of task-oriented training in patients with subacute stroke.