BIOACTIVITY OF ANTIDESMA BUNIUS LEAVES (EUPHORBIACEAE) AND THEIR MAJOR PHENOLIC CONSTITUENTS

Chromatographic fractionation of the methanol extract of Antidesma bunius leaves afforded six polyphenols, namely, corilagin (1), gallic (2), ferrulic (3) and ellagic (4) acids in addition to the flavone vicinin II (5) and the dimmer amentoflavone (6). Their structures were elucidated by NMR and HRESI analysis. This is the first reported occurrence of the biflavone (amentoflavone) and vicinin in this species. The total phenolic content was estimated at 90 mg/ml of gallic acid equivalent (GAE) per 100 g plant extract using the Folin-Ciocalteu method. Antioxidant biochemical assay of the plant methanol extract was carried out using culture medium of hepatocyte cell lines. An increase in glutathione reductase enzyme and a reduction of the nitric oxide level in the cell culture revealed that A. bunius leaf extract possesses significant antioxidant activity in comparing with quercetine as a reference. The hepatotoxicity and hepatoprotective activities of A.bunius leaf extract and the pure isolated compounds: 1 , 2 and 5 were evaluated by adopting the MTT colorimetric assay. Compounds 1 and 2 revealed a margin of safety on the monolayer hepatocyte with an IC50 > 1000 μg/ml while for 5, IC50 was at 125 μg/ml. The hepatoprotective activities for A.bunius leaf extract, 1 and 2 in comparison to Silymarin (50 μg/ml) were at 6.5, <12.5 and 12.5 μg/ml respectively while 5 did not show hepatoprotective activity at the tested concentrations

Transgelin is a TGFβ-inducible gene that regulates osteoblastic and adipogenic differentiation of human skeletal stem cells through actin cytoskeleston organization

Regenerative medicine is a novel approach for treating conditions in which enhanced bone regeneration is required. We identified transgelin (TAGLN), a transforming growth factor beta (TGFβ)-inducible gene, as an upregulated gene during in vitro osteoblastic and adipocytic differentiation of human bone marrow-derived stromal (skeletal) stem cells (hMSC). siRNA-mediated gene silencing of TAGLN impaired lineage differentiation into osteoblasts and adipocytes but enhanced cell proliferation. Additional functional studies revealed that TAGLN deficiency impaired hMSC cell motility and in vitro transwell cell migration. On the other hand, TAGLN overexpression reduced hMSC cell proliferation, but enhanced cell migration, osteoblastic and adipocytic differentiation, and in vivo bone formation. In addition, deficiency or overexpression of TAGLN in hMSC was associated with significant changes in cellular and nuclear morphology and cytoplasmic organelle composition as demonstrated by high content imaging and transmission electron microscopy that revealed pronounced alterations in the distribution of the actin filament and changes in cytoskeletal organization. Molecular signature of TAGLN-deficient hMSC showed that several genes and genetic pathways associated with cell differentiation, including regulation of actin cytoskeleton and focal adhesion pathways, were downregulated. Our data demonstrate that TAGLN has a role in generating committed progenitor cells from undifferentiated hMSC by regulating cytoskeleton organization. Targeting TAGLN is a plausible approach to enrich for committed hMSC cells needed for regenerative medicine application

Assessment of seismic scenario-structure based limit state criteria for a reinforced concrete high-rise building

Issues regarding reinforced concrete (RC) high-rise buildings involve different seismic scenarios, such as near- and far-field earthquakes, which can result in different levels and complex seismic scenario structures related to global damage. This study aims to evaluate the seismic scenario structures of a 23-storey RC high-rise building based on different damage measures and to develop a fragility curve with different limit state criteria. Six ground motions are selected to represent two seismic scenarios. Seismic scenario-based global response of the building at increasing earthquake intensity measures is adopted using incremental dynamic analysis (IDA). IDA curves and interstorey drift are used in the parametric study. The structural performance and damage measure of RC high-rise buildings are evaluated with performance-based seismic design limit state. The four performance levels proposed by ATC-43 are operational performance, immediate occupancy, life safety and collapse prevention. The IDA curves showed that near-field effect has a high frequency that gives impact at early intensity measures on building collapse compared with far-field effect. Meanwhile, interstorey drift result indicates that the near-field effect has a larger effect on building damages compared with the far-field effect. Based on the fragility curves, near-field earthquakes have a larger effect towards structural damages than far-field earthquakes

Detection of Biofilm Producing Staphylococci among Different Clinical Isolates and Its Relation to Methicillin Susceptibility

AIMS: To evaluate three in vitro phenotypic methods; tissue culture plate, tube method, and Congo red agar for detection of biofilm formation in staphylococci and assess the relation of biofilm formation with methicillin resistance and anti-microbial resistance. METHODS: The study included 150 staphylococcal isolates. Biofilm detection in staphylococci was performed using tissue culture plate, tube method, and Congo red agar. RESULTS: Tissue culture plate, tube method, and Congo red agar detected 74%, 42.7%, and 1.3% biofilm producing staphylococci respectively. S. aureus isolates were more common biofilm producers (53.2%) than CONS (46.8%). Biofilm production in CONS species was highest in S. hemolyticus (57.7%). Tube method was 51.4% sensitive, 82.1% specific. As for Congo red agar, sensitivity was very low (0.9%), but specificity was 97.4%. Biofilm producers were mostly; isolated from blood specimens (82.6%) and detected in methicillin-resistant strains 96/111 (86.5%). They were resistant to most antibiotics except vancomycin and linezolid. CONCLUSIONS: Tissue culture plate is a more quantitative and reliable method for detection of biofilm producing staphylococci compared to tube method and Congo red agar. Hence, it can still be used as a screening method for biofilm detection. Vancomycin and Linezolid are the most sensitive antibiotics among biofilm producing staphylococci

“In small places, close to home”: urban environmental impacts on child rights across four global cities

Urban environments influence child behaviours, exposures and experiences and may affect health, development, achievement and realization of fundamental human rights. We examined the status of eleven UN Convention on the Rights of the Child articles, in a multi-case study across four global cities. Within all study cities, children experienced unequal exposure to urban environmental risks and amenities. Many violations of child rights are related to car-based transportation systems and further challenged by pressures on urban systems from rapid population increases in the context of climate change. A child rights framework provides principles for a collective, multi-sectoral re-imagination of urban environments that support the human rights of all citizens

Traditional activities and general and mental health of adult Indigenous peoples living off-reserve in Canada

IntroductionWe examined associations between traditional Indigenous activities and self-perceived general and mental health in adult Indigenous persons living off-reserve in Canada using the 2012 and 2017 Aboriginal Peoples Surveys (APS), the two most recent datasets. We utilized four traditional Indigenous activities including hunting, making clothes or footwear, making arts or crafts, and gathering wild plants to investigate these self-reported data.MethodsData from 9,430 and 12,598 respondents from the 2012 and 2017 APS, respectively, who responded to 15 questions concerning traditional activities were assessed using multivariable logistic regression to produce odds ratios (OR) and 95% confidence intervals (CI). Covariates included age, sex, education-level, income-level, Indigenous identity, residential school connection, ability to speak an Indigenous language, smoking status, and alcohol consumption frequency.ResultsUsing the 2012 APS, clothes-making was associated with poor self-reported general (OR = 1.50, 95%CI: 1.12–1.99) and mental (OR = 1.59, 95%CI: 1.14–2.21) health. Hunting was associated with good mental health (OR = 0.71 95%CI: 0.56–0.93). Similarly, 2017 analyses found clothes-making associated with poor general health (OR = 1.25, 95%CI: 1.01–1.54), and hunting associated with good general (OR = 0.76, 95%CI: 0.64–0.89) and mental (OR = 0.69, 95%CI: 0.58–0.81) health. Artmaking was associated with poor general (OR = 1.37, 95%CI: 1.17–1.60) and mental (OR = 1.85, 95%CI: 1.58–2.17) health.ConclusionHunting had protective relationships with mental and general health, which may reflect benefits of participation or engagement of healthier individuals in this activity. Clothes-making and artmaking were associated with poor general and poor mental health, possibly representing reverse causation as these activities are often undertaken therapeutically. These findings have implications for future research, programs and policies concerning Indigenous health

Definition of Soybean Genomic Regions That Control Seed Phytoestrogen Amounts

Soybean seeds contain large amounts of isoflavones or phytoestrogens such as genistein, daidzein, and glycitein that display biological effects when ingested by humans and animals. In seeds, the total amount, and amount of each type, of isoflavone varies by 5 fold between cultivars and locations. Isoflavone content and quality are one key to the biological effects of soy foods, dietary supplements, and nutraceuticals. Previously we had identified 6 loci (QTL) controlling isoflavone content using 150 DNA markers. This study aimed to identify and delimit loci underlying heritable variation in isoflavone content with additional DNA markers. We used a recombinant inbred line (RIL) population ([Formula: see text]) derived from the cross of “Essex” by “Forrest,” two cultivars that contrast for isoflavone content. Seed isoflavone content of each RIL was determined by HPLC and compared against 240 polymorphic microsatellite markers by one-way analysis of variance. Two QTL that underlie seed isoflavone content were newly discovered. The additional markers confirmed and refined the positions of the six QTL already reported. The first new region anchored by the marker BARC_Satt063 was significantly associated with genistein ([Formula: see text] , [Formula: see text]) and daidzein ([Formula: see text] , [Formula: see text]). The region is located on linkage group B2 and derived the beneficial allele from Essex. The second new region defined by the marker BARC_Satt129 was significantly associated with total glycitein ([Formula: see text] , [Formula: see text]). The region is located on linkage group D1a+Q and also derived the beneficial allele from Essex. Jointly the eight loci can explain the heritable variation in isoflavone content. The loci may be used to stabilize seed isoflavone content by selection and to isolate the underlying genes

Genomic regions that underlie soybean seed isoflavone content

Soy products contain isoflavones (genistein, daidzein, and glycitein)that display biological effects when ingested by humans and animals, these effects are species, dose and age dependent. Therefore, the content and quality of isoflavones in soybeans is a key to their biological effect. Our objective was to identify loci that underlie isoflavone content in soybean seeds. The study involved 100 recombinant inbred lines (RIL)fr om the cross of ‘Essex’ by ‘Forrest,’ two cultivars that contrast for isoflavone content. Isoflavone content of seeds fromeach RIL was determined by high performance liquid chromatography (HPLC). The distribution of isoflavone content was continuous and unimodal. The heritability estimates on a line mean basis were 79% for daidzein, 22% for genistein, and 88% for glycitein. Isoflavone content of soybean seeds was compared against 150 polymorphic DNA markers in a one-way analysis of variance. Four genomic regions were found to be significantly associated with the isoflavone content of soybean seeds across both locations and years. Molecular linkage group B1 contained a major QTL underlying glycitein content (P = 0.0001,R2 = 50.2%), linkage group N contained a QTL for glycitein (P = 0.0033,R2 = 11.1%)and a QTL for daidzein (P = 0.0023,R2 = 10.3%) and linkage group A1 contained a QTL for daidzein (P = 0.0081,R2 = 9.6%). Selection for these chromosomal regions in a marker assisted selection program will allow for the manipulation of amounts and profiles of isoflavones (genistein, daidzein, and glycitein)c ontent of soybean seeds. In addition, tightly linked markers can be used in map based cloning of genes associated with isoflavone content

DEVELOPMENT OF NOVEL MICROWELL-PLATE SPECTROPHOTOMETRIC ASSAY FOR DETERMINATION OF VARENICLINE VIA ITS REACTION WITH CYCLOHEXA-3,5-DIENE-1,2-DIONE

This study describes, for the first, the spectrophotometric investigation for the condensation reaction between varenicline (VRC) and cyclohexa-3,5-diene-1,2-dione (CHDD). The reaction gave a violet-colored product exhibiting maximum absorption peak (λ max ) at 540 nm. The variables affecting the reaction were carefully investigated and the optimum conditions were established. The stoichiometry of the reaction was determined, and the reaction pathway was postulated. This color-developing reaction was employed in the development of microwell plate assay for VRC. In this assay, the reaction was carried out in 96-microwell plate and the absorbance of the colored-product was measured by microwell plate absorbance reader. Under the optimized reaction conditions, Beer's law correlating the absorbance with VRC concentration was obeyed in the range of 5 -100 µg/mL with good correlation coefficient (0.9986). The limits of detection and quantification were 2.29 and 6.95 µg/mL, respectively. The assay showed high precision as the values of relative standard deviations (RSD) did not exceed 2%. No interference was observed from the excipients that are present in VRC-containing tablets. The proposed assay was applied successfully for the determination of VRC in its pharmaceutical tablets with good accuracy and precisions; the label claim percentages were 98.80 ± 1.30%. The results were compared favorably with those of a reference prevalidated method. The proposed assay is practical and valuable in terms of its routine application in determination of VRC in in its bulk and tablets in pharmaceutical quality control laboratories

The histone H2B monoubiquitination regulatory pathway is required for differentiation of multipotent stem cells.

Extensive changes in posttranslational histone modifications accompany the rewiring of the transcriptional program during stem cell differentiation. However, the mechanisms controlling the changes in specific chromatin modifications and their function during differentiation remain only poorly understood. We show that histone H2B monoubiquitination (H2Bub1) significantly increases during differentiation of human mesenchymal stem cells (hMSCs) and various lineage-committed precursor cells and in diverse organisms. Furthermore, the H2B ubiquitin ligase RNF40 is required for the induction of differentiation markers and transcriptional reprogramming of hMSCs. This function is dependent upon CDK9 and the WAC adaptor protein, which are required for H2B monoubiquitination. Finally, we show that RNF40 is required for the resolution of the H3K4me3/H3K27me3 bivalent poised state on lineage-specific genes during the transition from an inactive to an active chromatin conformation. Thus, these data indicate that H2Bub1 is required for maintaining multipotency of hMSCs and plays a central role in controlling stem cell differentiation