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Sequencing-based genetic tests to identify individuals at increased risk of hereditary breast and ovarian cancers have resulted in the identification of more than 40,000 sequence variants of BRCA1 and BRCA2. A majority of these variants are considered to be variants of uncertain significance (VUS) because their impact on disease risk remains unknown, largely due to lack of sufficient familial linkage and epidemiological data. Several assays have been developed to examine the effect of VUS on protein function, which can be used to assess their impact on cancer susceptibility. In this study, we report the functional characterization of 88 BRCA2 variants, including several previously uncharacterized variants, using a well-established mouse embryonic stem cell (mESC)-based assay. We have examined their ability to rescue the lethality of Brca2 null mESC as well as sensitivity to six DNA damaging agents including ionizing radiation and a PARP inhibitor. We have also examined the impact of BRCA2 variants on splicing. In addition, we have developed a computational model to determine the probability of impact on function of the variants that can be used for risk assessment. In contrast to the previous VarCall models that are based on a single functional assay, we have developed a new platform to analyze the data from multiple functional assays separately and in combination. We have validated our VarCall models using 12 known pathogenic and 10 neutral variants and demonstrated their usefulness in determining the pathogenicity of BRCA2 variants that are listed as VUS or as variants with conflicting functional interpretation.

Significance Many neurons release neuropeptides as well as classical neurotransmitters. Neuropeptides often regulate neural circuits controlling behaviors such as emotional behaviors. However, how neuropeptide release is regulated is not well-understood. We identified a Drosophila gene, Dstac , that is similar to a vertebrate gene that regulates voltage-dependent calcium channels in skeletal muscles and found that it is active in muscles and a subset of neurons, including motor neurons. Using live imaging, electrophysiology, and genetic manipulations, we found that Dstac localizes to motor synapses with muscles and regulates the voltage response of calcium channels and the release of neuropeptides. Since Stac proteins are found in neurons in the vertebrate CNS, they may also be regulators of neuropeptide release in the vertebrate brain.

Purpose BRCA1 pathogenic variant heterozygotes are at a substantially increased risk for breast and ovarian cancer. The widespread uptake of testing has led to a significant increase in the detection of missense variants in BRCA1, the vast majority of which are variants of uncertain clinical significance (VUS), posing a challenge to genetic counseling. Here, we harness a wealth of functional data for thousands of variants to aid in variant classification. Methods We have collected, curated, and harmonized functional data for 2701 missense variants representing 24.5% of possible missense variants in BRCA1. Results were harmonized across studies by converting data into binary categorical variables (functional impact versus no functional impact). Using a panel of reference variants we identified a subset of assays with high sensitivity and specificity (≥80%) and apply the American College of Medical Genetics and Genomics/Association for Molecular Pathology (ACMG/AMP) variant interpretation guidelines to assign evidence criteria for classification. Results Integration of data from validated assays provided ACMG/AMP evidence criteria in favor of pathogenicity for 297 variants or against pathogenicity for 2058 representing 96.2% of current VUS functionally assessed. We also explore discordant results and identify limitations in the approach. Conclusion High quality functional data are available for BRCA1 missense variants and provide evidence for classification of 2355 VUS according to their pathogenicity.

Introduction: The biological green synthesis of nanoparticles via nanobiotechnology processes have a significant potential to boost nanoparticles production without the use of harsh, toxic, and expensive chemicals commonly used in conventional physical and chemical processes. Annona muricata, a tropical plant belonging to family Annonaceae is one of the most used plants in folk medicine because of its many medicinal uses and therefore presents a strong candidate for use in green synthesis. Aims: The aim of this study was to optimize a method for the synthesis of Silver Nanoparticles (AgNPs) from ethanolic extracts of leaves of Annona muricata as well as to characterize the green synthesized AgNPs. Methodology: AgNPs were synthesized from Annona muricata leaves using AgNO3 solution. The AgNPs were characterized using spectroscopy and microscopy techniques. Results: The formed AgNPs had an absorption maximum at 429 nm using UV–Visible spectroscopy and were stable under different pH, temperature, and storage conditions. Fourier transform infrared analysis revealed the different functional groups responsible for the synthesis and stabilization of the AgNPs. Scanning electron microscopy analysis revealed a spherical nature of the synthesized AgNPs. Energy dispersive x-ray spectroscopy analysis showed presence of Ag, O, and Cl with Ag having the highest composition at 60%. X-Ray Diffraction and Dynamic Light Scattering revealed a crystalline nature of AgNPs with an average size of 87.36 nm and a polydispersity index of 0.16 respectively. Transmission Electron Microscopy analysis further confirmed the crystalline and spherical nature of the AgNPs. Conclusion: In this article, an efficient, eco-friendly and low-cost method for the synthesis and recovery of stable AgNPs using Annona muricata leaves ethanolic extracts as both a reducing and capping agent has been reported for the first time. The synthesized AgNPs could be promising candidates for many biomedical, clinical, engineering, and polymer applications.

This research communication addresses the hypothesis that in dual-purpose goats, exposure to 1 h of extra-light given from 16 to 17 h after dawn (pulse of light) in winter stimulates milk yield. One group of goats was maintained under natural short photoperiod (natural day; ND (n = 7)). Another group of lactating females was submitted to an artificial long-day photoperiod consisting of 16 h light and 8 h darkness (long days; LD (n = 7)). A third group of females received one single hour of extra-light 16 h after the fixed dawn (pulse of light; PL (n = 6)). Goats from LD and PL yielded 30% more milk than goats from ND. Mean percentages of fat, protein and lactose contents in milk did not differ between the 3 groups at any stage of lactation, but these components in grams/day were higher in goats from PL than in the others two groups within the first 45 d of lactation. In conclusion, dual-purpose lactating goats that started their lactation during natural short days, the daily exposition to a 1-h pulse of light is sufficient to stimulate milk yield compared to females maintained under natural short photoperiod.

Green synthesis of nanoparticles from plant materials opens a new scope in nanobiotechnology and discourages the use of expensive toxic chemicals. The aim of this study was to develop and optimize a method for the synthesis of Silver Nanoparticles (AgNPs) from ethanolic extracts of fruits of Annona muricata as well as to characterize the green synthesized AgNPs. AgNPs were synthesized via AgNO3 solution. The AgNPs were characterized using spectroscopy and microscopy techniques. The formed AgNPs had an absorption maximum of 427 nm and were stable under different temperature, pH and storage conditions. Fourier Transform Infrared Resorption spectroscopy revealed the functional groups responsible for the synthesis and stabilization of the AgNPs. Scanning Electron Microscopy analysis revealed a spherical nature of the AgNPs. Energy Dispersive X-Ray spectroscopy showed presence of Ag, Cl, Ca, and Si with Ag having the highest composition at 80%. X-ray diffraction and dynamic light scattering revealed a crystalline nature of AgNPs with an average size of 60.12 nm and a polydispersity index of 0.1235 respectively. Transmission Electron Microscopy analysis further confirmed the crystalline and spherical nature of the AgNPs. In this article, an efficient, eco-friendly and low-cost method for the synthesis and recovery of stable AgNPs using ethanolic extracts of Annona muricata fruits as both reducing and capping agents has been reported. The synthesized AgNPs could have many biomedical and clinical applications.

Wild indigenous vegetables make considerable contributions to food baskets among subsistence farmers in sub‐Saharan Africa. The aim of this study was to evaluate the proximate analysis, mineral composition, vitamin C content, β‐carotene content, and GC‐MS profile of crude methanolic extracts of Asystasia mysorensis and Sesamum angustifolia. Crude extracts obtained through sequential extraction using ethyl acetate and methanol were screened for the presence of secondary metabolites. Functional groups present were determined with a Shimadzu FT‐IR spectrophotometer, while β‐carotene content and ascorbic acid content were evaluated using a Shimadzu HPLC and Shimadzu UV‐VIS spectrophotometer, respectively. Secondary metabolites present in the extracts were determined qualitatively using a Shimadzu GC‐MS system equipped with a NIST spectral database. From the results obtained, the two plants could supply the recommended daily requirement for micronutrient and vitamin C content needed for a healthy diet. The total phenolic and flavonoid contents in S. amgustifolia were higher as compared to A. myorensis; hence, their consumption is highly beneficial as some compounds identified in the GC‐MS profile have been reported to have medicinal properties. The findings on the mineral and chemical composition, GC‐MS profile of A. mysorensis and S. angustifolia indicate that their consumption may provide the recommended nutritional requirements needed for a healthy diet.

Genetic testing for BRCA1, a DNA repair protein, can identify carriers of pathogenic variants associated with a substantially increased risk for breast and ovarian cancers. However, an association with increased risk is unclear for a large fraction of BRCA1 variants present in the human population. Most of these variants of uncertain clinical significance lead to amino acid changes in the BRCA1 protein. Functional assays are valuable tools to assess the potential pathogenicity of these variants. Here, we systematically probed the effects of substitutions in the C terminus of BRCA1: the N- and C-terminal borders of its tandem BRCT domain, the BRCT-[N-C] linker region, and the α1 and α′1 helices in BRCT-[N] and -[C]. Using a validated transcriptional assay based on a fusion of the GAL4 DNA-binding domain to the BRCA1 C terminus (amino acids 1396–1863), we assessed the functional impact of 99 missense variants of BRCA1. We include the data obtained for these 99 missense variants in a joint analysis to generate the likelihood of pathogenicity for 347 missense variants in BRCA1 using VarCall, a Bayesian integrative statistical model. The results from this analysis increase our understanding of BRCA1 regions less tolerant to changes, identify functional borders of structural domains, and predict the likelihood of pathogenicity for 98% of all BRCA1 missense variants in this region recorded in the population. This knowledge will be critical for improving risk assessment and clinical treatment of carriers of BRCA1 variants.

Neuropeptides control rhythmic behaviors, but the timing and location of their release within circuits is unknown. Here imaging in the brain shows that synaptic neuropeptide release by Drosophila clock neurons is diurnal, peaking at times of day that were not anticipated by prior electrical and Ca2+ data. Furthermore, hours before peak synaptic neuropeptide release, neuropeptide release occurs at the soma, a neuronal compartment that has not been implicated in peptidergic transmission. The timing disparity between release at the soma and terminals results from independent and compartmentalized mechanisms for daily rhythmic release: consistent with conventional electrical activity-triggered synaptic transmission, terminals require Ca2+ influx, while somatic neuropeptide release is triggered by the biochemical signal IP3. Upon disrupting the somatic mechanism, the rhythm of terminal release and locomotor activity period are unaffected, but the number of flies with rhythmic behavior and sleep-wake balance are reduced. These results support the conclusion that somatic neuropeptide release controls specific features of clock neuron dependent behaviors. Thus, compartment specific mechanisms within individual clock neurons produce temporally and spatially partitioned neuropeptide release to expand the peptidergic connectome underlying daily rhythmic behaviors.Significance StatementIt is believed that electrical activity simultaneously stimulates widespread release sites in single neurons to elicit neuropeptide dependent behaviors. However, optically detecting neuropeptide release in the intact brain shows that clock neurons release neuropeptides from different sites at different times of the day. This is possible because one neuronal compartment, the soma, uses biochemical signaling instead of electrical activity to evoke release. Disrupting somatic release affects specific features of circadian locomotor activity and sleep. Thus, neuropeptide release is elicited by independent triggers from distinct parts of clock neurons to engage different regions of behavior regulating circuitry. This strategy for expanding the connectome may be used for other neuropeptide dependent behaviors, such as feeding and pain perception.

Summary Rotten food, maggots, bodily waste—all elicit disgust in humans. Disgust promotes survival by encouraging avoidance of disease vectors 1 but is also implicated in prejudice toward minority groups; avoidance of environmentally beneficial foods, such as insect protein; and maladaptive avoidance behavior in neuropsychiatric conditions. 2 , 3 , 4 , 5 Unlike fear, pathological disgust is not improved substantially by exposure therapy clinically, 6 nor in experimental work does behavioral avoidance of disgusting images habituate following prolonged exposure. 7 , 8 Under normal physiological conditions, perception of disgusting stimuli disrupts myoelectrical rhythms in the stomach, 9 , 10 , 11 , 12 , 13 inducing gastric dysrhythmias that correlate with neural signatures of disgust. 11 However, the causal role of gastric rhythm in disgust avoidance is unknown. We manipulated gastric rhythm using domperidone, a peripheral dopamine D2/D3 antagonist and common anti-emetic, at a dose (10 mg) that acts to convert gastric dysrhythmias to normal rhythms. 9 In a preregistered, randomized, double-blind, placebo-controlled crossover design in 25 healthy volunteers (aged 18–25), we measured the effects of domperidone on core disgust avoidance, using eye tracking to measure implicit (oculomotor) avoidance of disgusting images (feces) before and after an “exposure” intervention (monetary reinforcement for looking at disgusting images). 7 , 8 We find that domperidone significantly reduces oculomotor disgust avoidance following incentivized exposure. This suggests that domperidone may weaken the “immunity” of disgust to habituation, putatively by reducing gastric dysrhythmias during incentivized engagement with disgusting stimuli. This indicates a causal role for disgust-related visceral changes in disgust avoidance, supporting the hypothesis that physiological homeostasis contributes to emotional experience.

Huntington’s disease (HD), a genetic neurodegenerative disorder, primarily impacts the striatum and cortex with progressive loss of medium-sized spiny neurons (MSNs) and pyramidal neurons, disrupting cortico-striatal circuitry. A promising regenerative therapeutic strategy of transplanting human neural stem cells (hNSCs) is challenged by the need for long-term functional integration. We previously described that hNSCs transplanted into the striatum of HD mouse models differentiated into electrophysiologically active immature neurons, improving behavior and biochemical deficits. Here we show that 8-month implantation of hNSCs into the striatum of zQ175 HD mice ameliorates behavioral deficits, increases brain-derived neurotrophic factor (BDNF) and reduces mutant Huntingtin (mHTT) accumulation. Patch clamp recordings, immunohistochemistry and electron microscopy demonstrates that hNSCs differentiate into diverse neuronal populations, including MSN- and interneuron-like cells. Remarkably, hNSCs receive synaptic inputs, innervate host neurons, and improve membrane and synaptic properties. Overall, the findings support hNSC transplantation for further evaluation and clinical development for HD.

The genetic basis of Lewy body dementia (LBD) is not well understood. Here, we performed whole-genome sequencing in large cohorts of LBD cases and neurologically healthy controls to study the genetic architecture of this understudied form of dementia, and to generate a resource for the scientific community. Genome-wide association analysis identified five independent risk loci, whereas genome-wide gene-aggregation tests implicated mutations in the gene GBA. Genetic risk scores demonstrate that LBD shares risk profiles and pathways with Alzheimer's disease and Parkinson's disease, providing a deeper molecular understanding of the complex genetic architecture of this age-related neurodegenerative condition.

SUMMARYAging is an intricate process that is characterized by multiple hallmarks including stem cell exhaustion, genome instability, epigenome alteration, impaired proteostasis, and cellular senescence. While each of these traits is detrimental at the cellular level, it remains unclear how they are interconnected to cause systemic organ deterioration. Here we show that abrogating Brap, a BRCA1 associated protein important for neurogenesis, results in cellular senescence with persistent DNA double-strand breaks and elevation of histone H2A mono- and poly-ubiquitination (H2Aub). The high H2Aub initiates histone proteolysis, leading to both epigenetic alteration and proteasome overflow. These defects induce neuroinflammation, impair proteostasis, accelerate neurodegeneration, and substantially shorten lifespan in mice carrying Brap deletions in the brain. We further show H2Aub is also increased in human brain tissues of Alzheimer’s disease. These data together suggest that chromatin aberrations mediated by H2Aub act as a nexus of multiple aging hallmarks and promote tissue-wide degeneration.

SummaryFor a century [1,2], studies of monozygotic and dizygotic twins have yielded estimates of trait heritability. The clever logic behind them is that while both types of twins share environments, their genetic overlap is different. Hence, larger trait correlations between monozygotic compared to dizygotic twins indicate heritability (nature), whereas similar correlations indicate shared environmental influences (nurture), and low correlations indicate shaping through non-shared environments (external influences and measurement error). While many have written on the assumptions that both types of twins share equal environments [3–5], and that parental genetics and environment are independent [6,7]; fewer have put their data where their mouth is. Here, the impacts of unmet assumptions were investigated using a generative mixture model of twin phenotypes. The results indicated that violations of the equal environments assumption yielded large overestimations of heritability and underestimations of shared environmental influences. On the other hand, when parental genetics shaped twins’ shared environments, only minor non-linear biases against heritability emerged. Finally, realistic levels of measurement error uniformly depressed estimates for genetic and shared environmental factors. In sum, twin studies are particularly susceptible to overestimation of genetic and non-shared environmental influences. This bias could explain why some traits, such as attitudes towards property taxes [8], show suspiciously high heritability without a biologically plausible mechanism. Particularly in the context of traits with convincing mechanisms of cultural transmission [9–11] and complex gene-environment interactions [6], researchers should not allow biases in twin studies to overestimate heritability.

By replacing lost or dysfunctional myocardium, tissue regeneration is a promising approach to treat heart failure. However, the challenge of detecting bona fide heart regeneration limits the validation of potential regenerative factors. One method to detect new cardiomyocytes is multicolor lineage tracing with clonal analysis. Clonal analysis experiments can be difficult to undertake, because labeling conditions that are too sparse lack sensitivity for rare events such as cardiomyocyte proliferation, and diffuse labeling limits the ability to resolve clones. Presented here is a protocol to undertake clonal analysis of the neonatal mouse heart by using statistical modeling of nearest neighbor distributions to resolve cardiomyocyte clones. This approach enables resolution of clones over a range of labeling conditions and provides a robust analytical approach for quantifying cardiomyocyte proliferation and regeneration. This protocol can be adapted to other tissues and can be broadly used to study tissue regeneration.

Tissue patterning by gradients of form-generating substances, or ‘morphogens,’ is a fundamental paradigm of developmental biology. Nonetheless, it has been unclear whether classic morphogen gradients pattern the mammalian telencephalon and neocortex, or whether these highly specialized and evolved tissues use entirely unique mechanisms. Recent studies indicate that, like most other tissues, the telencephalon is indeed patterned by classic morphogen gradients. This chapter reviews the history and general principles of morphogen gradients in development, the known sources of morphogens in the developing telencephalon, and the evidence that bone morphogenetic proteins and fibroblast growth factors act as graded morphogens to pattern the dorsal telencephalon and neocortex. It then highlights the unique and complex interactions among morphogens within the dorsal telencephalon, and the roles of morphogen gradients and their interactions in human developmental anomalies, particularly holoprosencephaly.

Many variants of uncertain significance (VUS) have been identified in BRCA2 through clinical genetic testing. VUS pose a significant clinical challenge because the contribution of these variants to cancer risk has not been determined. We conducted a comprehensive assessment of VUS in the BRCA2 C-terminal DNA binding domain (DBD) by using a validated functional assay of BRCA2 homologous recombination (HR) DNA-repair activity and defined a classifier of variant pathogenicity. Among 139 variants evaluated, 54 had ≥99% probability of pathogenicity, and 73 had ≥95% probability of neutrality. Functional assay results were compared with predictions of variant pathogenicity from the Align-GVGD protein-sequence-based prediction algorithm, which has been used for variant classification. Relative to the HR assay, Align-GVGD significantly (p < 0.05) over-predicted pathogenic variants. We subsequently combined functional and Align-GVGD prediction results in a Bayesian hierarchical model (VarCall) to estimate the overall probability of pathogenicity for each VUS. In addition, to predict the effects of all other BRCA2 DBD variants and to prioritize variants for functional studies, we used the endoPhenotype-Optimized Sequence Ensemble (ePOSE) algorithm to train classifiers for BRCA2 variants by using data from the HR functional assay. Together, the results show that systematic functional assays in combination with in silico predictors of pathogenicity provide robust tools for clinical annotation of BRCA2 VUS.

The Huntington’s disease protein Huntingtin (Htt) regulates axonal transport of dense-core vesicles (DCVs) containing neurotrophins and neuropeptides. DCVs travel down axons to reach nerve terminals where they are either captured in synaptic boutons to support later release or reverse direction to reenter the axon as part of vesicle circulation. Currently, the impact of Htt on DCV dynamics in the terminal is unknown. Here we report that knockout of Drosophila Htt selectively reduces retrograde DCV flux at proximal boutons of motoneuron terminals. However, initiation of retrograde transport at the most distal bouton and transport velocity are unaffected suggesting that synaptic capture rate of these retrograde DCVs could be altered. In fact, tracking DCVs shows that retrograde synaptic capture efficiency is significantly elevated by Htt knockout or knockdown. Furthermore, synaptic boutons contain more neuropeptide in Htt knockout larvae even though bouton size, single DCV fluorescence intensity, neuropeptide release in response to electrical stimulation and subsequent activity-dependent capture are unaffected. Thus, loss of Htt increases synaptic capture as DCVs travel by retrograde transport through boutons resulting in reduced transport toward the axon and increased neuropeptide in the terminal. These results therefore identify native Htt as a regulator of synaptic capture and neuropeptide storage.

Summary Transplantation of neural stem cells (NSCs) can improve cognition in animal models of Alzheimer's disease (AD). However, AD is a protracted disorder, and prior studies have examined only short-term effects. We therefore used an immune-deficient model of AD (Rag-5xfAD mice) to examine long-term transplantation of human NSCs (StemCells Inc.; HuCNS-SCs). Five months after transplantation, HuCNS-SCs had engrafted and migrated throughout the hippocampus and exhibited no differences in survival or migration in response to β-amyloid pathology. Despite robust engraftment, HuCNS-SCs failed to terminally differentiate and over a quarter of the animals exhibited ectopic human cell clusters within the lateral ventricle. Unlike prior short-term experiments with research-grade HuCNS-SCs, we also found no evidence of improved cognition, no changes in brain-derived neurotrophic factor, and no increase in synaptic density. These data, while disappointing, reinforce the notion that individual human NSC lines need to be carefully assessed for efficacy and safety in appropriate long-term models., Highlights • Human neural stem cells (HuCNS-SC) have been used in multiple human clinical trials • HuCNS-SC originally derived under GMP conditions did not improve cognition in AD mice • HuCNS-SC failed to differentiate, improve synaptic density, or increase BDNF levels • HuCNS-SC formed ectopic ventricular clusters in a quarter of transplanted mice, In this article, Blurton-Jones and colleagues demonstrate that StemCells, Inc. human neural stem cells (HuCNS-SC), which were originally derived under GMP conditions, failed to improve cognition, synaptic density, or increase BDNF in an immune-deficient AD mouse model. Furthermore, cells formed ectopic ventricular clusters in over a quarter of transplanted animals.

Anaerobic microorganisms of the Geobacter genus are effective electron sources for the synthesis of nanoparticles, for bioremediation of polluted water, and for the production of electricity in fuel cells. In multistep reactions, electrons are transferred via iron/heme cofactors of c‐type cytochromes from the inner cell membrane to extracellular metal ions, which are bound to outer membrane cytochromes. We measured electron production and electron flux rates to 5×105 e s−1 per G. sulfurreducens. Remarkably, these rates are independent of the oxidants, and follow zero order kinetics. It turned out that the microorganisms regulate electron flux rates by increasing their Fe2+/Fe3+ ratios in the multiheme cytochromes whenever the activity of the extracellular metal oxidants is diminished. By this mechanism the respiration remains constant even when oxidizing conditions are changing. This homeostasis is a vital condition for living systems, and makes G. sulfurreducens a versatile electron source.

Green synthesized metallic nanoparticles are environmentally friendly, bio-compatible, and highly stable. The aim of this study was to synthesize iron nanoparticles (FeNPs) from FeCl3 solution using aqueous leaf extracts of Galinsoga parviflora (Gp), Conyza bonariensis (Cb) and Bidens pilosa (Bp) and use them in degradation of methylene blue dye. The iron nanoparticles were characterized using UV–Vis spectrophotometer, FT-IR spectrophotometer, X-ray Fluorescence (EDXRF), X-ray diffractometer (XRD), and scanning electron microscope (SEM). Phytochemical screening for presence of secondary metabolites revealed presence of phenolics, phytosterols and flavonoids. The total phenolic and flavonoid content in Galinsoga parviflora, Conyza bonariensis and Bidens pilosa leaf extracts were 57.67 ± 1.27, 117.13 ± 0.03, 126.27 ± 0.013 mg Gallic Equivalent/g of Dry Weight (mg GAE/g DW) and 39.00 ± 0.56, 45.50 ± 0.59, 33.13 ± 0.81 mg Rutin Equivalent/g of Dry Weight (mg RE/g DW) respectively. The UV–Vis spectrum of FeCl3 had a shoulder at 320 nm, which disappeared upon addition of G. parviflora, C. bonariensis and B. pilosa extracts confirming formation of iron nanoparticles. Evaluation of iron content of the synthesized nanoparticles revealed that the iron content in G. parviflora, C. bonariensis and B. pilosa extracts was 51, 47 and 44% respectively. XRD data revealed presence of a large amorphous coating that masked iron peaks, though 2 theta values observed have been reported to be of iron oxides. Methylene blue degradation studies revealed that CbNPs, BpNPs and GpNPs synthesized were good biocatalysts as they degraded the dye by 86, 84.3 and 92% respectively. Therefore, green synthesized iron nanoparticles is cost effective and environmentally safe in providing insight in the environmental removal of dyes.

In this data article, data obtained from an efficient, eco-friendly and low-cost method for the synthesis and recovery of Silver nanoparticles (AgNPs) using ethanolic extracts of Annona muricata fruits and leaves as reducing, stabilizing and capping agents has been reported. 99.7% pure silver nitrate was used as the inorganic ion source. The data was obtained using different spectroscopic and microscopic techniques. The data is presented in form of images, Microsoft excel sheets, graphs,.raw files,.dpt files, PDF files, among others. Methods of analysis and interpretation of the data have also been presented. The data can be most useful to researchers, research students, industrialists and academicians to acquire knowledge on the green synthesis of AgNPs and related applications. The data is deposited in the Mendeley Data Repository as two independent datasets accessible at https://doi.org/10.17632/jkj2x782wh.1 Gavamukulya et al., 2019 and https://doi.org/10.17632/f4mb6b488n.1 Gavamukulya et al., 2019. Keywords: Annona muricata, Green synthesis, Silver nanoparticles (AgNPs), UV/VIS, FTIR, XRD

Metallic nanoparticles synthesized using aqueous plant extracts are environment-friendly, biocompatible, and highly stable. The aim of this study was to synthesize iron nanoparticles using aqueous Ageratum conyzoides extracts and evaluating their antimicrobial and photocatalytic properties. The particles were analysed using UV–Vis spectrophotometer, FT-IR Spectrophotometer, X-ray diffractometer and Scanning electron microscope. GC–MS profile of the extracts revealed presence of secondary metabolites which were further quantified to determine the total phenolic and total flavonoids content of the extracts. The antibacterial activity of the plant extract and the synthesized iron oxide nanoparticles was evaluated against five microorganisms using agar well diffusion method. Iron nanoparticles synthesized in a one step process observed using visible spectra and the functional groups present such as C=O were identified from IR spectrum. SEM–EDX profile identified presence of iron, oxygen, chlorine, calcium in the particles while XRD data revealed the particles synthesized were composed oxides of iron which had moderate activity against the selected microorganisms as compared to the antibiotic ciprofloxacin. The particles were able to photocatalytic degrade methylene blue with a degradation efficiency of 92%. The results obtained in this study confirms that Ageratum conyzoides can play an important role in the bioreduction of Fe ions to FeNPs which have moderate activity against microorganisms and can act as photocatalyst to degrade methylene blue.

Background: Germline DNA copy number variation (CNV) is a ubiquitous source of genetic variation and remains largely unexplored in association with epithelial ovarian cancer (EOC) risk. Methods: CNV was quantified in the DNA of approximately 3,500 cases and controls genotyped with the Illumina 610k and HumanOmni2.5M arrays. We performed a genome-wide association study of common (>1%) CNV regions (CNVRs) with EOC and high-grade serous (HGSOC) risk and, using The Cancer Genome Atlas (TCGA), performed in silico analyses of tumor-gene expression. Results: Three CNVRs were associated (P < 0.01) with EOC risk: two large (∼100 kb) regions within the 610k set and one small ( Conclusions: Though CNV appears to not contribute largely to EOC susceptibility, a number of low-to-common frequency variants may influence the risk of EOC and tumor-gene expression. Impact: Further research on CNV and EOC susceptibility is warranted, particularly with CNVs estimated from high-density arrays.

Neuronal cholinergic circuits have been implicated in cognitive function and neurological disease, but the role of cholinergic signaling in other cellular populations within the brain has not been as fully defined. Here, we show that cholinergic signaling mechanisms are involved in mediating the function of the choroid plexus, the brain structure responsible for generating CSF and releasing various factors into the brain. The choroid plexus was found to express markers of endogenous cholinergic signaling, including multiple nicotinic acetylcholine receptor (nAChR) subtypes in a region-specific manner, and application of nicotine was found to induce cellular activation, as evidenced by calcium influx in primary tissue. During intravenous nicotine self-administration in male rats, nicotine increased expression of transthyretin, a protein selectively produced and released by the choroid plexus, and microRNA-204 (mir-204), a transcript found in high levels in the choroid plexus and CSF. Finally, human choroid plexus tissue from both sexes was found to exhibit similar nAChR, transthyretin and mir-204 expression profiles, supporting the translational relevance of the findings. Together, these studies demonstrate functionally active cholinergic signaling mechanisms in the choroid plexus, the resulting effects on transthyretin and mir-204 expression, and reveal the direct mechanism by which nicotine modulates function of this tissue.

Many neurons influence their targets through co-release of neuropeptides and small-molecule transmitters. Neuropeptides are packaged into dense-core vesicles (DCVs) in the soma and then transported to synapses, while small-molecule transmitters such as monoamines are packaged by vesicular transporters that function at synapses. These separate packaging mechanisms point to activity, by inducing co-release as the sole scaler of co-transmission. Based on screening in Drosophila for increased presynaptic neuropeptides, the receptor protein tyrosine phosphatase (Rptp) Ptp4E was found to post-transcriptionally regulate neuropeptide content in single DCVs at octopamine synapses. This occurs without changing neuropeptide release efficiency, transport and DCV size measured by both stimulated emission depletion super-resolution and transmission electron microscopy. Ptp4E also controls the presynaptic abundance and activity of the vesicular monoamine transporter (VMAT), which packages monoamine transmitters for synaptic release. Thus, rather than rely on altering electrical activity, the Rptp regulates packaging underlying monoamine-neuropeptide co-transmission by controlling vesicular membrane transporter and luminal neuropeptide content.This article has an associated First Person interview with the first author of the paper.

Genome-wide association studies have identified 40 ovarian cancer risk loci. However, the mechanisms underlying these associations remain elusive. In this study, we conducted a two-pronged approach to identify candidate causal SNPs and assess underlying biological mechanisms at chromosome 9p22.2, the first and most statistically significant associated locus for ovarian cancer susceptibility. Three transcriptional regulatory elements with allele-specific effects and a scaffold/matrix attachment region were characterized and, through physical DNA interactions, BNC2 was established as the most likely target gene. We determined the consensus binding sequence for BNC2 in vitro, verified its enrichment in BNC2 ChIP-seq regions, and validated a set of its downstream target genes. Fine-mapping by dense regional genotyping in over 15,000 ovarian cancer cases and 30,000 controls identified SNPs in the scaffold/matrix attachment region as among the most likely causal variants. This study reveals a comprehensive regulatory landscape at 9p22.2 and proposes a likely mechanism of susceptibility to ovarian cancer. Significance: Mapping the 9p22.2 ovarian cancer risk locus identifies BNC2 as an ovarian cancer risk gene. See related commentary by Choi and Brown, p. 439

IntroductionSex hormone binding globulin (SHBG) levels are reported to be inversely associated with diabetes risk. It is unknown whether diabetes prevention interventions increase SHBG and whether resultant changes in SHBG affect diabetes risk. The purpose of this analysis was to determine whether intensive lifestyle intervention (ILS) or metformin changed circulating SHBG and if resultant changes influenced diabetes risk in the Diabetes Prevention Program (DPP).Research design and methodsThis is a secondary analysis from the DPP (1996–2001), a randomized trial of ILS or metformin versus placebo on diabetes risk over a mean follow-up of 3.2 years. The DPP was conducted across 27 academic study centers in the USA. Men, premenopausal and postmenopausal women without hormone use in the DPP were evaluated. The DPP included overweight/obese persons with elevated fasting glucose and impaired glucose tolerance. Main outcomes measures were changes in SHBG levels at 1 year and risk of diabetes over 3 years.ResultsILS resulted in significantly higher increases (postmenopausal women: pConclusionsLifestyle intervention may be associated with favorable changes in circulating SHBG, which is largely due to changes in adiposity. Changes in circulating SHBG do not independently predict reductions in diabetes incidence.

Synaptic neuropeptide and neurotrophin stores are maintained by constitutive bidirectional capture of dense-core vesicles (DCVs) as they circulate in and out of the nerve terminal. Activity increases DCV capture to rapidly replenish synaptic neuropeptide stores following release. However, it is not known whether this is due to enhanced bidirectional capture. Here experiments at theDrosophilaneuromuscular junction, where DCVs contain neuropeptides and a bone morphogenic protein, show that activity-dependent replenishment of synaptic neuropeptides following release is evident after inhibiting the retrograde transport with the dynactin disruptor mycalolide B or photobleaching DCVs entering a synaptic bouton by retrograde transport. In contrast, photobleaching anterograde transport vesicles entering a bouton inhibits neuropeptide replenishment after activity. Furthermore, tracking of individual DCVs moving through boutons shows that activity selectively increases capture of DCVs undergoing anterograde transport. Finally, upregulating fragile X mental retardation 1 protein (Fmr1, also called FMRP) acts independently of futsch/MAP-1B to abolish activity-dependent, but not constitutive, capture. Fmr1 also reduces presynaptic neuropeptide stores without affecting activity-independent delivery and evoked release. Therefore, presynaptic motoneuron neuropeptide storage is increased by a vesicle capture mechanism that is distinguished from constitutive bidirectional capture by activity dependence, anterograde selectivity, and Fmr1 sensitivity. These results show that activity recruits a separate mechanism than used at rest to stimulate additional synaptic capture of DCVs for future release of neuropeptides and neurotrophins.SIGNIFICANCE STATEMENTSynaptic release of neuropeptides and neurotrophins depends on presynaptic accumulation of dense-core vesicles (DCVs). At rest, DCVs are captured bidirectionally as they circulate throughDrosophilamotoneuron terminals by anterograde and retrograde transport. Here we show that activity stimulates further synaptic capture that is distinguished from basal capture by its selectivity for anterograde DCVs and its inhibition by overexpression of the fragile X retardation protein Fmr1. Fmr1 dramatically lowers DCV numbers in synaptic boutons. Therefore, activity-dependent anterograde capture is a major determinant of presynaptic peptide stores.

Multiphoton autofluorescence shows that NAD(P)H is elevated in dopamine neurons. Much of the NAD(P)H is localized to the ER, and perturbing mitochondrial function alters ER NAD(P)H. The NAD(P)H coupling between neuronal ER and mitochondria provides a means for the two organelles to coordinate their redox stress responses., Pyridine nucleotides are redox coenzymes that are critical in bioenergetics, metabolism, and neurodegeneration. Here we use brain slice multiphoton microscopy to show that substantia nigra dopamine neurons, which are sensitive to stress in mitochondria and the endoplasmic reticulum (ER), display elevated combined NADH and NADPH (i.e., NAD(P)H) autofluorescence. Despite limited mitochondrial mass, organellar NAD(P)H is extensive because much of the signal is derived from the ER. Remarkably, even though pyridine nucleotides cannot cross mitochondrial and ER membranes, inhibiting mitochondrial function with an uncoupler or interrupting the electron transport chain with cyanide (CN−) alters ER NAD(P)H. The ER CN− response can occur without a change in nuclear NAD(P)H, raising the possibility of redox shuttling via the cytoplasm locally between neuronal mitochondria and the ER. We propose that coregulation of NAD(P)H in dopamine neuron mitochondria and ER coordinates cell redox stress signaling by the two organelles.

Complement system activation is recognized as a deleterious component of the mammalian physiological response to traumatic injury with severe hemorrhage (TH). Female Yorkshire swine were subjected to a simulated austere prehospital battlefield scenario. Each animal underwent controlled hemorrhage of 22 mL/kg at 100 mL/min rate for approximately 10 min followed by soft tissue injury, femur fracture, and spleen injury. Subsequent blood loss was uncontrolled. Twenty-eight minutes postinjury the animals were randomized into treatment or no treatment with recombinant human C1 esterase inhibitor (C1INH) (500 IU/kg, n = 11) and into receiving or not permissive hypotensive resuscitation (n = 14) with infusion of 45 mL/kg lactated Ringer's solution (2× blood lost). Observation and animal maintenance continued for 6 h at which time the animals had either expired or were euthanized. Heart, lung, and small intestine tissue samples were collected. Pharmacokinetic, hemodynamic, and metabolic parameters as well as survival time, plasma complement activity and tissue deposition, cytokine levels, and tissue injury were determined. We found that administration of C1INH protected tissues from damage, reduced the levels of inflammatory cytokines, and improved blood chemistry. Immunohistochemical analyses revealed that C1INH administration following TH markedly reduced complement activation and deposition in tissues. Importantly, C1INH administration prolonged survival of animals particularly in those which received resuscitation fluid infusion. Our data urge early administration of C1INH to limit organ damage and prolong survival of those injured in the battlefield.

A Drosophila model system is used to show that the hereditary spastic paraplegia proteins spastin and atlastin help axons but not dendrites regenerate. The endoplasmic reticulum concentrates at tips of regenerating axons but not dendrites, and this depends on spastin and atlastin., Mutations in >50 genes, including spastin and atlastin, lead to hereditary spastic paraplegia (HSP). We previously demonstrated that reduction of spastin leads to a deficit in axon regeneration in a Drosophila model. Axon regeneration was similarly impaired in neurons when HSP proteins atlastin, seipin, and spichthyin were reduced. Impaired regeneration was dependent on genetic background and was observed when partial reduction of HSP proteins was combined with expression of dominant-negative microtubule regulators, suggesting that HSP proteins work with microtubules to promote regeneration. Microtubule rearrangements triggered by axon injury were, however, normal in all genotypes. We examined other markers to identify additional changes associated with regeneration. Whereas mitochondria, endosomes, and ribosomes did not exhibit dramatic repatterning during regeneration, the endoplasmic reticulum (ER) was frequently concentrated near the tip of the growing axon. In atlastin RNAi and spastin mutant animals, ER accumulation near single growing axon tips was impaired. ER tip concentration was observed only during axon regeneration and not during dendrite regeneration. In addition, dendrite regeneration was unaffected by reduction of spastin or atlastin. We propose that the HSP proteins spastin and atlastin promote axon regeneration by coordinating concentration of the ER and microtubules at the growing axon tip.

Stem Cell Reports Letters human subjects, nor become, in the absence of safety con- cerns, the exclusive factor on which clinical testing is based. In closing, we wish to express our gratitude to the physicians and patients who understand the nature of clinical translation and that human outcomes ulti- mately determine whether therapeutic testing continues to advance. REFERENCES Anderson, A.J., Piltti, K.M., Hooshmand, M.J., Nishi, R.A., and Cum- mings, B.J. (2017). Stem Cell Rep. 8, this issue, 249–263. Marsh, S.E., Yeung, S.T., Torres, M., Lau, L., Davis, J.L., Monuki, E.S., Poon, W.W., and Blurton-Jones, M. (2017). Stem Cell Rep. 8, this issue, 235–248. Response to StemCells Inc. Edwin S. Monuki, 1,2,3 Aileen J. Anderson, 1,2,4,5 Mathew Blurton-Jones, 1,2,6, * and Brian J. Cummings 1,2,4,5, * 1 Sue & Bill Gross Stem Cell Research Center for Memory Impairments & Neurological Disorders 3 Department of Pathology & Laboratory Medicine 4 Department of Physical & Medical Rehabilitation 5 Department of Anatomy & Neurobiology 6 Department of Neurobiology & Behavior University of California-Irvine, Irvine, CA 92697, USA *Correspondence: mblurton@uci.edu (M.B.-J.), cummings@uci.edu (B.J.C.) http://dx.doi.org/10.1016/j.stemcr.2017.02.002 2 Institute We welcome the opportunity for further discussion of this data, and in the paragraphs below, we briefly address the comments provided by StemCells Inc. (STEM). One issue raised is that the data in both papers encompass development of novel therapeutics for conditions with no effective therapy. We agree that the need to move clinical testing forward for conditions in which there are no effec- tive therapeutics cannot be overstated. Cervical spinal cord injury (SCI) and Alzheimer’s disease (AD) are both prime examples of this need, although with some critical differ- ences. As many reviews have summarized, risk tolerance in progressing to a clinical trial is inherently different for terminal conditions in comparison with chronic condi- tions, and different again for chronic conditions involving greater or lesser impacts on the activities of daily living or quality of life. We believe that publication of these studies supports this need by opening further discussion of trans- lational research, including its pitfalls. In the case of cell therapeutics, and as discussed in the primary papers (An- derson et al., 2017; Marsh et al., 2017), we suggest that these include testing of the final clinical product, careful consideration of the tenant of informed consent for the subjects that choose to enroll in a clinical trial of any kind, and the relationship between pre-clinical research and clinical trial success rate. A parallel issue in the clinical development of a cell therapy for SCI is disclosure of pre-clinical animal study outcomes to clinical investigators for the company’s Pathway Study and to the US FDA in regulatory filings. We, of course, agree that full disclosure and open discus- sion are critical to the integrity of conducting a clinical trial, but we have no direct knowledge of what exactly was disclosed to clinical investigators or in regulatory fil- ings for the ‘‘Pathway’’ cervical SCI trial, or when. In this regard, the timeline as we know is as follows. A pre- liminary analysis of the findings detailed in the Anderson et al. (2017) was provided to STEM in the form of a Research Performance Progress Report for the associated U01 on July 1, 2014. The failure of the CCL to meet the U01 efficacy milestone in the cervical model was made clear and resulted in early termination of U01 funding. A more complete analysis of these data was submitted to STEM in the form of a set of presentation slides on December 3, 2014. Transplantation of the first patient in the Pathway study was announced December 18, 2014. A face-to-face meeting was held between the SCI team and STEM on January 22, 2015. However, in the absence of further information, this timeline provides rather limited insight. The key issues would seem to be what in- formation was shared and in what form, and whether it can be ethically considered that informed consent was achieved for the subjects enrolling in the trial (Anderson and Cummings, 2016). It is understood that the invest- ment and intellectual property of a sponsor requires Stem Cell Reports j Vol. 8 j 194–197 j February 14, 2017 j a 2017 The Author(s). 195 This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Large predators play important ecological roles, yet many are disproportionately imperiled. In marine systems, artificial reefs are often deployed to restore degraded reefs or supplement existing reefs, but it remains unknown whether these interventions benefit large predators. Comparative field surveys of thirty artificial and natural reefs across ~200 km of the North Carolina, USA coast revealed large reef-associated predators were more dense on artificial than natural reefs. This pattern was associated with higher densities of transient predators (e.g. jacks, mackerel, barracuda, sharks) on artificial reefs, but not of resident predators (e.g., grouper, snapper). Further analyses revealed that this pattern of higher transient predator densities on artificial reefs related to reef morphology, as artificial reefs composed of ships hosted higher transient predator densities than concrete reefs. The strength of the positive association between artificial reefs and transient predators increased with a fundamental habitat trait–vertical extent. Taller artificial reefs had higher densities of transient predators, even when accounting for habitat area. A global literature review of high trophic level fishes on artificial and natural habitats suggests that the overall pattern of more predators on artificial habitats is generalizable. Together, these findings provide evidence that artificial habitats, especially those like sunken ships that provide high vertical structure, may support large predators.

Thymidylate synthase (TYMS) is a crucial enzyme for DNA synthesis. TYMS expression is regulated by its antisense mRNA, ENOSF1. Disrupted regulation may promote uncontrolled DNA synthesis and tumor growth. We sought to replicate our previously reported association between rs495139 in the TYMS-ENOSF1 3′ gene region and increased risk of mucinous ovarian carcinoma (MOC) in an independent sample. Genotypes from 24,351 controls to 15,000 women with invasive OC, including 665 MOC, were available. We estimated per-allele odds ratios (OR) and 95% confidence intervals (CI) using unconditional logistic regression, and meta-analysis when combining these data with our previous report. The association between rs495139 and MOC was not significant in the independent sample (OR = 1.09, 95% CI = 0.97–1.22, p = 0.15, N = 665 cases). Meta-analysis suggested a weak association (OR = 1.13, 95% CI = 1.03–1.24, p = 0.01, N = 1019 cases). No significant association with risk of other OC histologic types was observed (p = 0.05 for tumor heterogeneity). In expression quantitative trait locus (eQTL) analysis, the rs495139 allele was positively associated with ENOSF1 mRNA expression in normal tissues of the gastrointestinal system, particularly esophageal mucosa (r = 0.51, p = 1.7 × 10−28), and nonsignificantly in five MOC tumors. The association results, along with inconclusive tumor eQTL findings, suggest that a true effect of rs495139 might be small.

Background Choroid plexus epithelial cells express high levels of transthyretin, produce cerebrospinal fluid and many of its proteins, and make up the blood-cerebrospinal fluid barrier. Choroid plexus epithelial cells are vital to brain health and may be involved in neurological diseases. Transgenic mice containing fluorescent and luminescent reporters of these cells would facilitate their study in health and disease, but prior transgenic reporters lost expression over the early postnatal period. Methods Human bacterial artificial chromosomes in which the transthyretin coding sequence was replaced with DNA for tdTomato or luciferase 2 were used in pronuclear injections to produce transgenic mice. These mice were characterized by visualizing red fluorescence, immunostaining, real-time reverse transcription polymerase chain reaction, and luciferase enzyme assay. Results Reporters were faithfully expressed in cells that express transthyretin constitutively, including choroid plexus epithelial cells, retinal pigment epithelium, pancreatic islets, and liver. Expression of tdTomato in choroid plexus began at the appropriate embryonic age, being detectable by E11.5. Relative levels of tdTomato transcript in the liver and choroid plexus paralleled relative levels of transcripts for transthyretin. Expression remained robust over the first postnatal year, although choroid plexus transcripts of tdTomato declined slightly with age whereas transthyretin remained constant. TdTomato expression patterns were consistent across three founder lines, displayed no sex differences, and were stable across several generations. Two of the tdTomato lines were bred to homozygosity, and homozygous mice are healthy and fertile. The usefulness of tdTomato reporters in visualizing and analyzing live Transwell cultures was demonstrated. Luciferase activity was very high in homogenates of choroid plexus and continued to be expressed through adulthood. Luciferase also was detectable in eye and pancreas. Conclusions Transgenic mice bearing fluorescent and luminescent reporters of transthyretin should prove useful for tracking transplanted choroid plexus epithelial cells, for purifying the cells, and for reporting their derivation from stem cells. They also should prove useful for studying transthyretin synthesis by other cell types, as transthyretin has been implicated in many functions and conditions, including clearance of β-amyloid peptides associated with Alzheimer’s disease, heat shock in neurons, processing of neuropeptides, nerve regeneration, astrocyte metabolism, and transthyretin amyloidosis.

Background: Plants are new sources of antibacterial agents, hence the need to determine and evaluate the antibacterial properties, antioxidant activity and gas chromatography – mass spectrometer (GC-MS) profile of medicinal plants. Methodology: In this study, sequential extraction of Prunus africana and Harrisonia abyssinica was used to obtain ethyl acetate and methanol extracts. Antioxidant activity was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH), whereas the total phenolic and total flavonoid contents were estimated using Folin-Ciocalteu and aluminium chloride, respectively. Antibacterial properties of the extracts against Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli and Candida albicans were estimated using the disc diffusion method and compared against streptomycin. Results: Screening of crude methanolic extracts revealed the presence of secondary metabolites which was further confirmed by Fourier transform infrared (FT-IR) characterisation that revealed the presence of H-bonded OH functional groups. The extracts revealed that P. africana had a higher total phenolic and total flavonoid contents compared to H. abyssinica . Methanolic extracts of both plants had moderate activity against selected microorganisms and both inhibited DPPH radical scavenging activity. GC-MS analysis of P. africana and H. abyssinica extracts revealed the presence of several phytochemicals that have been reported to have medicinal uses. Total phenolic and flavonoid contents showed positive correlations with the DPPH radical scavenging activity and negative correlations with EC 50 . Conclusion: Prunus africana and H. abyssinica extracts had moderate antimicrobial properties against the selected microorganisms because of the presence of secondary metabolites.

Background: Increased resistance against antimicrobial medication used to combat bacterial infection necessitates the need for alternative medication. Objective: This study seeks to evaluate the antimicrobial potential and phytochemical profiles of 10 medicinal plants collected from Chuka, Tharaka-Nithi County of Kenya. Plant samples were collected, dried, pulverised into fine powders and extracted with distilled water. Methodology: Qualitative phytochemical screening and disc-agar diffusion methods were used according to standard method to determine phytochemical profile and antimicrobial activity of the aqueous extracts against four bacterial strains and a fungus. Results: Phytochemical investigation revealed the presence of terpenoids, tannins and reducing sugars in all the plants except one. Saponins were found to be present in Albizia anthelmintica, Entada leptostachya, Raponae rhododendroides and Warbugia ugandensis . Steroids were present in seven plants while alkaloids were present in five. Albizia anthelmintica, E. leptostachya and W. ugandensis extracts were active against Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli and Candida albicans. Escherichia coli was the most susceptible bacteria against all the plant extracts tested, except Harrisonia abyssinica. Vernonia lasiopus and Uvariodendron anisatum were the least active extracts. Susceptibility against E. coli and C. albicans was significantly comparable to benzathine penicillin and streptomycin. Conclusion: The type of ailments the plants are claimed to treat can be attributed to the presence of various classes of phytochemicals. In conclusion, the plants evaluated were found to be active against the microorganisms tested.

Background: Natural phytoconstituents produced by plants for their sustenance have been reported to reduce disease. Objectives: This study determined the phytoconstituents, antioxidant and antimicrobial activity of crude methanolic extracts of Entada leptostachya and Prosopis juliflora extracts. Methodology: Antioxidant activity was determined using 2,2-diphenyl-1-picrylhydrazyl and β-carotene assays; the total phenolic and flavonoid were estimated using Folin–Ciocalteau and aluminium chloride, whereas antimicrobial activity was determined using the zone of inhibition method. Results: Screening of the extracts revealed the presence of terpenoids, flavonoids, saponins and phenols. Fourier transform infrared spectra of the extracts revealed presence of hydrogen bonded –OH functional group. E. leptostachya barks had the highest antioxidant activity followed by P. juliflora roots, E. leptostachya roots and P. juliflora leaves [μg/mL]. Prosopis juliflora (roots) had the highest bleaching effect, whereas E. leptostachya (barks) had the lowest bleaching effect. The total flavonoids were determined to be 0.15 ± 0.02 mg/g, 1.18 ± 0.18 mg/g, 0.39 ± 0.05 mg/g and 0.64 ± 0.03 mg/g for E. leptostachya roots, E. leptostachya barks, P. juliflora leaves and P. juliflora roots extracts, respectively. The total phenols were determined to be 0.93 ± 0.18 mg/g, 2.69 ± 0.41 mg/g, 0.62 ± 0.08 mg/g and 0.62 ± 0.08 mg/g for E. leptostachya roots, E. leptostachya barks, P. juliflora roots and P. juliflora leaves extracts. All plant extracts exhibited moderate activity against the growth of selected microorganisms. Conclusion: Antimicrobial and antioxidant activity of the two plants was as a result of secondary metabolites found in the crude extracts.

Epithelial ovarian cancer (EOC) is the fifth leading cause of cancer mortality in American women. Normal ovarian physiology is intricately connected to small GTP binding proteins of the Ras superfamily (Ras, Rho, Rab, Arf, and Ran) which govern processes such as signal transduction, cell proliferation, cell motility, and vesicle transport. We hypothesized that common germline variation in genes encoding small GTPases is associated with EOC risk. We investigated 322 variants in 88 small GTPase genes in germline DNA of 18,736 EOC patients and 26,138 controls of European ancestry using a custom genotype array and logistic regression fitting log-additive models. Functional annotation was used to identify biofeatures and expression quantitative trait loci that intersect with risk variants. One variant, ARHGEF10L (Rho guanine nucleotide exchange factor 10 like) rs2256787, was associated with increased endometrioid EOC risk (OR = 1.33, p = 4.46 x 10-6). Other variants of interest included another in ARHGEF10L, rs10788679, which was associated with invasive serous EOC risk (OR = 1.07, p = 0.00026) and two variants in AKAP6 (A-kinase anchoring protein 6) which were associated with risk of invasive EOC (rs1955513, OR = 0.90, p = 0.00033; rs927062, OR = 0.94, p = 0.00059). Functional annotation revealed that the two ARHGEF10L variants were located in super-enhancer regions and that AKAP6 rs927062 was associated with expression of GTPase gene ARHGAP5 (Rho GTPase activating protein 5). Inherited variants in ARHGEF10L and AKAP6, with potential transcriptional regulatory function and association with EOC risk, warrant investigation in independent EOC study populations. ispartof: PLOS ONE vol:13 issue:7 ispartof: location:United States status: published

Background: Genome-wide association studies (GWAS) have so far reported 12 loci associated with serous epithelial ovarian cancer (EOC) risk. We hypothesized that some of these loci function through nearby transcription factor (TF) genes and that putative target genes of these TFs as identified by coexpression may also be enriched for additional EOC risk associations. Methods: We selected TF genes within 1 Mb of the top signal at the 12 genome-wide significant risk loci. Mutual information, a form of correlation, was used to build networks of genes strongly coexpressed with each selected TF gene in the unified microarray dataset of 489 serous EOC tumors from The Cancer Genome Atlas. Genes represented in this dataset were subsequently ranked using a gene-level test based on results for germline SNPs from a serous EOC GWAS meta-analysis (2,196 cases/4,396 controls). Results: Gene set enrichment analysis identified six networks centered on TF genes (HOXB2, HOXB5, HOXB6, HOXB7 at 17q21.32 and HOXD1, HOXD3 at 2q31) that were significantly enriched for genes from the risk-associated end of the ranked list (P < 0.05 and FDR < 0.05). These results were replicated (P < 0.05) using an independent association study (7,035 cases/21,693 controls). Genes underlying enrichment in the six networks were pooled into a combined network. Conclusion: We identified a HOX-centric network associated with serous EOC risk containing several genes with known or emerging roles in serous EOC development. Impact: Network analysis integrating large, context-specific datasets has the potential to offer mechanistic insights into cancer susceptibility and prioritize genes for experimental characterization. Cancer Epidemiol Biomarkers Prev; 24(10); 1574–84. ©2015 AACR.

Epithelial-mesenchymal transition (EMT) is a process whereby epithelial cells assume mesenchymal characteristics to facilitate cancer metastasis. However, EMT also contributes to the initiation and development of primary tumors. Prior studies that explored the hypothesis that EMT gene variants contribute to epithelial ovarian carcinoma (EOC) risk have been based on small sample sizes and none have sought replication in an independent population. We screened 15,816 single-nucleotide polymorphisms (SNPs) in 296 genes in a discovery phase using data from a genome-wide association study of EOC among women of European ancestry (1,947 cases and 2,009 controls) and identified 793 variants in 278 EMT-related genes that were nominally (P < 0.05) associated with invasive EOC. These SNPs were then genotyped in a larger study of 14,525 invasive-cancer patients and 23,447 controls. A P-value