STABILITY-INDICATING LIQUID CHROMATOGRAPHY METHOD FOR THE DETERMINATION OF PENTOXYVERINE CITRATE AND ITS DEGRADANT

Objectives: The objectives of this research were to develop and validate a HPLC method for determination of Pentoxyverine citrate (PNX) and its degradant (DG). Methods: Forced degradation studies were performed on bulk sample using alkaline (0.1M sodium hydroxide) and acid (1M hydrochloric acid. The proposed method was based on using a 150 x 4.6 mm (i. d.) (Luna, Phenomenex, Torrance, CA, USA) (5 µm particle size) reversed phase C18 column with mobile phase consisting of a mixture of methanol-10 mM sodium dihydrogen phosphate pH 4 in ratio of (60:40, v/v) and UV detection at 230 nm with flow rate of 1 mL min-1. Results: The linear calibration range was between 10-40 mg ml-1and 10-40 mg ml-1for PNX and DG respectively. The method was found to be accurate with 100.23% and 100.07% recovery for PNX and DG respectively. The limit of detection (LOD) was found to be 3.79 x10-2μg ml-1 and 4.24 x10-2μg ml-1 for PNX and DG respectively, while the limit of quantification (LOQ) was found to be 12.62 x10-2μg ml-1 and 14.12 x10-2μg ml-1 for PNX and DG respectively. PNX was found to be most stable at a pH of 5.7. Conclusion: The validation study of the proposed method was successfully carried out and the method was found to be suitable and economic for routine determination of PNX in pharmaceutical syrup, without any interference from the excipients, and in the presence of its acidic and alkaline degradation products

Effect of zinc supplementation on growth Hormone Insulin growth factor axis in short Egyptian children with zinc deficiency

BACKGROUND: The relationship between zinc (Zn) and growth hormone-insulin growth factor (GH-IGF) system and how Zn therapy stimulates growth in children has not been clearly defined in humans. Thus, we aimed to assess GH-IGF axis in short children with Zn deficiency and to investigate the effect of Zn supplementation on these parameters. METHODS: Fifty pre-pubertal Egyptian children with short stature and Zn deficiency were compared to 50 age-, sex-, and pubertal stage- matched controls. All subjects were subjected to history, auxological assessment and measurement of serum Zn, IGF-1, insulin growth factor binding protein-3 (IGFBP-3); and basal and stimulated GH before and 3 months after Zn supplementation (50 mg/day). RESULTS: After 3 months of Zn supplementation in Zn-deficient patients, there were significant increases in height standard deviation score (SDS, P = 0.033), serum Zn (P < 0.001), IGF-1 (P < 0.01), IGF-1 standard deviation score (SDS,P < 0.01) and IGFBP-3 (P = 0.042). Zn rose in all patients but reached normal ranges in 64 %, IGF-1 levels rose in 60 % but reached normal ranges in 40 % and IGFBP-3 levels rose in 40 % but reached reference ranges in 22 %. Growth velocity (GV) SDS did not differ between cases and controls (p = 0.15) but was higher in GH-deficient patients than non-deficient ones, both having Zn deficiency (p = 0.03). CONCLUSION: Serum IGF-1 and IGFBP-3 levels were low in short children with Zn deficiency, and increased after Zn supplementation for 3 months but their levels were still lower than the normal reference ranges in most children; therefore, Zn supplementation may be necessary for longer periods

Noggin depletion in adipocytes promotes obesity in mice.

ObjectiveObesity has increased to pandemic levels and enhanced understanding of adipose regulation is required for new treatment strategies. Although bone morphogenetic proteins (BMPs) influence adipogenesis, the effect of BMP antagonists such as Noggin is largely unknown. The aim of the study was to define the role of Noggin, an extracellular BMP inhibitor, in adipogenesis.MethodsWe generated adipose-derived progenitor cells and a mouse model with adipocyte-specific Noggin deletion using the AdiponectinCre transgenic mouse, and determined the adipose phenotype of Noggin-deficiency.ResultsOur studies showed that Noggin is expressed in progenitor cells but declines in adipocytes, possibly allowing for lipid accumulation. Correspondingly, adipocyte-specific Noggin deletion in&nbsp;vivo promoted age-related obesity in both genders with no change in food intake. Although the loss of Noggin caused white adipose tissue hypertrophy, and whitening and impaired function in brown adipose tissue in both genders, there were clear gender differences with the females being most affected. The females had suppressed expression of brown adipose markers and thermogenic genes including peroxisome proliferator activated receptor gamma coactivator 1 alpha (PGC1alpha) and uncoupling protein 1 (UCP1) as well as genes associated with adipogenesis and lipid metabolism. The males, on the other hand, had early changes in a few BAT markers and thermogenic genes, but the main changes were in the genes associated with adipogenesis and lipid metabolism. Further characterization revealed that both genders had reductions in VO2, VCO2, and RER, whereas females also had reduced heat production. Noggin was also reduced in diet-induced obesity in inbred mice consistent with the obesity phenotype of the Noggin-deficient mice.ConclusionsBMP signaling regulates female and male adipogenesis through different metabolic pathways. Modulation of adipose tissue metabolism by select BMP antagonists may be a strategy for long-term regulation of age-related weight gain and obesity

EFFECT OF ASCORBATE, SALICYLATE AND SILICATE ON POTATO PLANT UNDER WATER DEFICIT STRESS CONDITIONS

The effect of different concentrations of ascorbate (As) at 0, 0.25, 0.5 and 1.0 mM, salicylate (Sa) at 0, 0.25, 0.5 and 1.0 mM , in addition to silicate (Si) at 0, 100, 250, and 500 ppm on potato plant subjected to different water regimes; three days interval (control), four days interval (4DI) and five days interval (5DI).  Photosynthetic pigments, free proline and malonyldialdehyde (MDA) concentrations, and plant dry matter percentage were determined. Also, tuber yield was recorded and the percentage of tuber dry matter was determined. Water deficit resulted in significant reduction in tuber yield, plant dry matter percentage beside significant increase in free proline and MDA contents. Meanwhile, effects on photosynthetic pigments content and tuber dry matter were inconsistent. The effects of using different concentrations of ascorbate (As) under water deficit provide a superior effect of As at 0.25 mM. Data revealed that ascorbate treatment enhanced tuber yield by improved Chl (a and b) and carotenoids content, with a promoted reduction in free proline content and MDA. Salicylate (Sa) treatment under water deficit exhibited partial enhancement in photosynthetic pigments; where, Sa (1.0 mM) increased Chl a under all water regimes and Chl b at control and 4DI water regimes. While Sa (0.5 mM) improved carotenoids content at 5DI water regimes. The positive effect of salicylate on tuber yield was obvious only at 5DI water regime; as salicylate concentrations (0.25, 0.5 and 1.0 mM) enhanced tuber yield by 65.61, 21.8 and 33.33%. Data exhibited that these increments were concomitant with a  reduction in free proline content and a slight increase in tuber dry matter. Interactive effects of Silicate (Si)/water deficit were variable with the concentration under the used water deficit regimes, where, Si (100 ppm), showed higher means of Chl (a and b) with a reduction in free proline contents, at both 4DI and 5DI. The higher tuber yield resulted only at 5DI. More pronounced effect, Si (250 ppm) exhibited increment of photosynthetic pigments, plant dry matter, tuber yield, along with slight reduction in MDA, particularly at 5DI. It could be concluded that silicate treatment alleviated water stress by improving carotenoids content, plant dry matter and reducing both free proline and MDA contents, which attributes enhancement in tuber yield under water deficit. Generally, lower concentrations of As, Sa and Si showed their benefits under higher water stress regimes

Dominance and G×E interaction effects improvegenomic prediction and genetic gain inintermediate wheatgrass (Thinopyrumintermedium)

Genomic selection (GS) based recurrent selection methods were developed to accelerate the domestication of intermediate wheatgrass [IWG, Thinopyrum intermedium (Host) Barkworth & D.R. Dewey]. A subset of the breeding population phenotyped at multiple environments is used to train GS models and then predict trait values of the breeding population. In this study, we implemented several GS models that investigated the use of additive and dominance effects and G×E interaction effects to understand how they affected trait predictions in intermediate wheatgrass. We evaluated 451 genotypes from the University of Minnesota IWG breeding program for nine agronomic and domestication traits at two Minnesota locations during 2017–2018. Genet-mean based heritabilities for these traits ranged from 0.34 to 0.77. Using fourfold cross validation, we observed the highest predictive abilities (correlation of 0.67) in models that considered G×E effects. When G×E effects were fitted in GS models, trait predictions improved by 18%, 15%, 20%, and 23% for yield, spike weight, spike length, and free threshing, respectively. Genomic selection models with dominance effects showed only modest increases of up to 3% and were trait-dependent. Crossenvironment predictions were better for high heritability traits such as spike length, shatter resistance, free threshing, grain weight, and seed length than traits with low heritability and large environmental variance such as spike weight, grain yield, and seed width. Our results confirm that GS can accelerate IWG domestication by increasing genetic gain per breeding cycle and assist in selection of genotypes with promise of better performance in diverse environments

Cathelicidin suppresses lipid accumulation and hepatic steatosis by inhibition of the CD36 receptor.

Background and objectivesObesity is a global epidemic which increases the risk of the metabolic syndrome. Cathelicidin (LL-37 and mCRAMP) is an antimicrobial peptide with an unknown role in obesity. We hypothesize that cathelicidin expression correlates with obesity and modulates fat mass and hepatic steatosis.Materials and methodsMale C57BL/6 J mice were fed a high-fat diet. Streptozotocin was injected into mice to induce diabetes. Experimental groups were injected with cathelicidin and CD36 overexpressing lentiviruses. Human mesenteric fat adipocytes, mouse 3T3-L1 differentiated adipocytes and human HepG2 hepatocytes were used in the in vitro experiments. Cathelicidin levels in non-diabetic, prediabetic and type II diabetic patients were measured by enzyme-linked immunosorbent assay.ResultsLentiviral cathelicidin overexpression reduced hepatic steatosis and decreased the fat mass of high-fat diet-treated diabetic mice. Cathelicidin overexpression reduced mesenteric fat and hepatic fatty acid translocase (CD36) expression that was reversed by lentiviral CD36 overexpression. Exposure of adipocytes and hepatocytes to cathelicidin significantly inhibited CD36 expression and reduced lipid accumulation. Serum cathelicidin protein levels were significantly increased in non-diabetic and prediabetic patients with obesity, compared with non-diabetic patients with normal body mass index (BMI) values. Prediabetic patients had lower serum cathelicidin protein levels than non-diabetic subjects.ConclusionsCathelicidin inhibits the CD36 fat receptor and lipid accumulation in adipocytes and hepatocytes, leading to a reduction of fat mass and hepatic steatosis in vivo. Circulating cathelicidin levels are associated with increased BMI. Our results demonstrate that cathelicidin modulates the development of obesity

Genomic epidemiology of the first epidemic wave of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in Palestine.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the novel coronavirus responsible for the COVID-19 pandemic, continues to cause a significant public-health burden and disruption globally. Genomic epidemiology approaches point to most countries in the world having experienced many independent introductions of SARS-CoV-2 during the early stages of the pandemic. However, this situation may change with local lockdown policies and restrictions on travel, leading to the emergence of more geographically structured viral populations and lineages transmitting locally. Here, we report the first SARS-CoV-2 genomes from Palestine sampled from early March 2020, when the first cases were observed, through to August of 2020. SARS-CoV-2 genomes from Palestine fall across the diversity of the global phylogeny, consistent with at least nine independent introductions into the region. We identify one locally predominant lineage in circulation represented by 50 Palestinian SARS-CoV-2, grouping with genomes generated from Israel and the UK. We estimate the age of introduction of this lineage to 05/02/2020 (16/01/2020-19/02/2020), suggesting SARS-CoV-2 was already in circulation in Palestine predating its first detection in Bethlehem in early March. Our work highlights the value of ongoing genomic surveillance and monitoring to reconstruct the epidemiology of COVID-19 at both local and global scales

IL-4Rα on dendritic cells in neonates and Th2 immunopathology in respiratory syncytial virus infection

© Society for Leukocyte Biology. Respiratory syncytial virus (RSV) is one of the leading causes of bronchiolitis in children, and severe RSV infection early in life has been associated with asthma development. Using a neonatal mouse model, we have shown that down-regulation of IL-4 receptor α (IL-4Rα) with antisense oligonucleotides in the lung during neonatal infection protected from RSV immunopathophysiology. Significant down-regulation of IL-4Rα was observed on pulmonary CD11b+ myeloid dendritic cells (mDCs) suggesting a role for IL-4Rα on mDCs in the immunopathogenesis of neonatal RSV infection. Here, we demonstrated that neonatal CD11b+ mDCs expressed higher levels of IL-4Rα than their adult counterparts. Because CD11b+ mDCs mainly present antigens to CD4+ T cells, we hypothesized that increased expression of IL- 4Rα on neonatal CD11b+ mDCs was responsible for Th2 - biased RSV immunopathophysiology. Indeed, when IL-4Rα was selectively deleted from CD11b+ mDCs, the immunopathophysiology typically observed following RSV reinfection was ablated, including Th2 inflammation, airway-mucus hyperproduction, and pulmonary dysfunction. Further, overexpression of IL-4Rα on adult CD11b+ DCs and their adoptive transfer into adult mice was able to recapitulate the Th2-biased RSV immunopathology typically observed only in neonates infected with RSV. IL-4Rα levels on CD11c+ cells were inversely correlated with maturation status of CD11b+ mDCs upon RSV infection. Our data demonstrate that developmentally regulated IL-4Rα expression is critical for the maturity of pulmonary CD11b+ mDCs and the Th2-biased immunopathogenesis of neonatal RSV infection

Molecular marker dissection of stem rust resistance in Nebraska bread wheat germplasm

Stem rust (caused by Puccinia graminis f. sp. tritici) is a major disease of wheat. To understand the genetic basis of stem rust resistance in Nebraska winter wheat, a set of 330 genotypes representing two nurseries (DUP2015 and TRP2015) were evaluated for resistance to a Nebraska stem rust race (QFCSC) in two replications. The TRP2015 nursery was also evaluated for its resistance to an additional 13 stem rust races. The analysis of variance revealed significant variation among genotypes in both populations for stem rust resistance. Nine stem rust genes, Sr6, Sr31, Sr1RSAmigo, Sr24, Sr36, SrTmp, Sr7b, Sr9b, and Sr38, were expected and genotyped using gene-specific markers. The results of genetic analysis confirmed the presence of seven stem rust resistance genes. One genotype (NE15680) contained target alleles for five stem rust resistance genes and had a high level of stem rust resistance against different races. Single marker analysis indicated that Sr24 and Sr38 were highly significantly associated with stem rust resistance in the DUP2015 and TRP2015 nurseries, respectively. Linkage disequilibrium analysis identified the presence of 17 SNPs in high linkage with the Sr38-specific marker. These SNPs potentially tagging the Sr38 gene could be used in marker-assisted selection after validating them in additional genetic backgrounds

Egyptian truffles as a source of antimicrobial and antioxidant agents

Edible desert truffles are obligate symbiotic macro hypogenous ascomycetes fruit bodies. These are growing in depth between 5 and 10 cm. These consider a miracle of nature and an unexploited source of therapeutic compounds with antimicrobial, antioxidant, and wonderful food, especially for Bedouins. The current study investigated the activity of premature and mature ethyl acetate truffle extracts (white and Red) collected from the Western Egyptian Desert as antimicrobial and antioxidant. This study evaluated in vitro the efficacy of antimicrobial activity of organic truffles extract, and their effect on various pathogens (Gram-negative, Gram-positive bacteria, filamentous fungi, and yeast) by using agar well diffusion. TEM micrographs had been done for the most effective crude extracts. Furthermore, the activity of DPPH scavenging was studied for both mature truffles. Both truffles extracts had antibacterial activity more than antifungal activity. The selected extracts exhibited an inhibitory effect on the cell wall and protoplasm of pathogens. Terfezia sp. had antioxidant activity more than Tirmania sp. This investigation concluded that the truffle extracts could be considered a promising antibiotic and antioxidant drug in near future