The relationship between DNA methyltransferase 3B (DNMT3B) and miR 124-3pa expressions in bladder cancer tissues.

BACKGROUND: Cancer bladder is the most common malignant tumor affecting the urinary tract. Genetic alterations are tightly associated with the development of cancer bladder. MicroRNAs (miRNA) are small, noncoding single-stranded RNA molecules that have been linked to bladder cancer. miR-124-3pa exhibits altered expression in various types of human malignancies. DNA methyltransferase 3B (DNMT3B) is responsible for de novo DNA methylation which is a fundamental epigenetic process in carcinogenesis. This work was performed to study the expression of DNMT3B and miR 124-3pa in bladder cancer tissues, and investigate their significance in the diagnosis and prognosis of the disease. SUBJECTS & METHODS: This case-control study included one hundred and six tissue samples of patients with primary urothelial bladder cancer. The tissues were separated into two parts. The first part was immediately frozen and kept at - 80 °C for total RNA extraction with subsequent detection of miR 124-3pa and DNMT3B expressions. The other part was preserved in formalin solution for histopathological examination. RESULTS: There was a highly statistically significant difference between the cancerous and the normal tissues as regarding miRNA-124-3pa and DNMT3B expression (P < 0.001) for each. Also, there was a highly statistically significant strong negative correlation between miRNA-124-3pa and DNMT3B expression (r=-0.750, P < 0.001). The combined performance of miR-124-3pa and DNMT3B revealed that the cutoff point of ≥ 3.3 can be used as a predictor of the presence of cancer bladder with sensitivity of 98.1% and specificity of 80%. CONCLUSION: miR-124-3pa and DNMT3B can be used as predictors of the presence of cancer bladder.

Integrative application of heavy metal-resistant bacteria, moringa extracts, and nano-silicon improves spinach yield and declines its contaminant contents on a heavy metal-contaminated soil.

Microorganism-related technologies are alternative and traditional methods of metal recovery or removal. We identified and described heavy metal-resistant bacteria isolated from polluted industrial soils collected from various sites at a depth of 0-200 mm. A total of 135 isolates were screened from polluted industrial soil. The three most abundant isolate strains resistant to heavy metals were selected: Paenibacillus jamilae DSM 13815T DSM (LA22), Bacillus subtilis ssp. spizizenii DSM 15029T DSM (MA3), and Pseudomonas aeruginosa A07_08_Pudu FLR (SN36). A test was conducted to evaluate the effect of (1) isolated heavy metal-resistant bacteria (soil application), (2) a foliar spray with silicon dioxide nanoparticles (Si-NPs), and (3) moringa leaf extract (MLE) on the production, antioxidant defense, and physio-biochemical characteristics of spinach grown on heavy metal-contaminated soil. Bacteria and MLE or Si-NPs have been applied in single or combined treatments. It was revealed that single or combined additions significantly increased plant height, shoot dry and fresh weight, leaf area, number of leaves in the plant, photosynthetic pigments content, total soluble sugars, free proline, membrane stability index, ascorbic acid, relative water content, α-tocopherol, glycine betaine, glutathione, and antioxidant enzyme activities (i.e., peroxidase, glutathione reductase, catalase, superoxide dismutase, and ascorbate peroxidase) compared with the control treatment. However, applying bacteria or foliar spray with MLE or Si-NPs significantly decreased the content of contaminants in plant leaves (e.g., Fe, Mn, Zn, Pb, Cd, Ni, and Cu), malondialdehyde, electrolyte leakage, superoxide radical ( O 2 · - ) , and hydrogen peroxide (H2O2). Integrative additions had a more significant effect than single applications. It was suggested in our study that the integrative addition of B. subtilis and MLE as a soil application and as a foliar spray, respectively, is a critical approach to increasing spinach plant performance and reducing its contaminant content under contaminated soil conditions.

A case of acheiria.

Congenital limb anomalies are rare. Acheiria is a congenital limb abnormality that presents as an absence of the hand and it is often diagnosed by prenatal ultrasonography. Herein we present a case of an 11-year-old female patient with acheiria. This case stresses on further studying the relationship between advanced pregnancy age, drugs or herb use during pregnancy and acheiria.

Screening of Bioactive Compounds from Endophytic Marine-Derived Fungi in Saudi Arabia: Antimicrobial and Anticancer Potential.

Nowadays, endophytic fungi represent a rich source of biological active compounds. In the current study, twelve endophytic fungal species were isolated from Avicennia marina leaves. From the isolates, Aspergillus niger, Penicillium rubens and Alternaria alternata recorded the highest isolation frequency (80%), relative density (12.5%) and antimicrobial activity. The antimicrobial and anticancer activities of P. rubens were more effective than those of A. niger and A. alternata; therefore, its identification was confirmed via the ITS rRNA gene. Filtrate extracts of P. rubens, A. alternata and A. niger were analyzed using GC-MS and showed different detected constituents, such as acetic acid ethyl ester, N-(4,6-Dimethyl-2-pyrimidinyl)-4-(4-nitrobenzylideneamino) benzenesulfonamide, 1,2-benzenedicarboxylic acid, hexadecanoic acid and octadecanoic acid. Filtrate extract of P. rubens exhibited the presence of more compounds than A. alternata and A. niger. Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and Aspergillus fumigatus were more inhibited by P. rubens extract than A. alternata or A. niger, with inhibition zones of 27.2 mm, 22.21 mm, 26.26 mm, 27.33 mm, 28.25 mm and 8.5 mm, respectively. We observed negligible cytotoxicity of P. rubens extract against normal cells of human lung fibroblasts (WI-38 cell line), unlike A. alternata and A. niger extracts. Proliferation of prostate cancer (PC-3) was inhibited using P. rubens extract, exhibiting mortality levels of 75.91% and 76.2% at 200 µg/mL and 400 µg/mL of the extract. Molecular docking studies against the crystal structures of C. albicans (6TZ6) and the cryo-EM structure of B. subtilis (7CKQ) showed significant interactions with benzenedicarboxylic acid and N-(4,6-dimethyl-2-pyrimidinyl)-4-(4-nitrobenzylideneamino) benzenesulfonamide as a constituent of P. rubens extract. N-(4,6-dimethyl-2-pyrimidinyl)-4-(4-nitrobenzylideneamino) benzenesulfonamide had the highest scores of −6.04905 kcal/mol and −6.590 kcal/mol towards (6tz6) and (7CKQ), respectively.

Global Patterns and Drivers of Soil Dissimilatory Nitrate Reduction to Ammonium.

Dissimilatory nitrate reduction to ammonium (DNRA), the nearly forgotten process in the terrestrial nitrogen (N) cycle, can conserve N by converting the mobile nitrate into non-mobile ammonium avoiding nitrate losses via denitrification, leaching, and runoff. However, global patterns and controlling factors of soil DNRA are still only rudimentarily known. By a meta-analysis of 231 observations from 85 published studies across terrestrial ecosystems, we find a global mean DNRA rate of 0.31 ± 0.05 mg N kg-1 day-1, being significantly greater in paddy soils (1.30 ± 0.59) than in forests (0.24 ± 0.03), grasslands (0.52 ± 0.15), and unfertilized croplands (0.18 ± 0.04). Soil DNRA was significantly enhanced at higher altitude and lower latitude. Soil DNRA was positively correlated with precipitation, temperature, pH, soil total carbon, and soil total N. Precipitation was the main stimulator for soil DNRA. Total carbon and pH were also important factors, but their effects were ecosystem-specific as total carbon stimulates DNRA in forest soils, whereas pH stimulates DNRA in unfertilized croplands and paddy soils. Higher temperatures inhibit soil DNRA via decreasing total carbon. Moreover, nitrous oxide (N2O) emissions were negatively related to soil DNRA. Thus, future changes in climate and land-use may interact with management practices that alter soil substrate availability and/or soil pH to enhance soil DNRA with positive effects on N conservation and lower N2O emissions.

Kidney dysfunction requiring dialysis is a heterogeneous syndrome: we should treat it like one.

PURPOSE OF REVIEW: Advanced kidney failure requiring dialysis, commonly labeled end-stage kidney disease or chronic kidney disease stage 5D, is a heterogeneous syndrome -a key reason that may explain why: treating advanced kidney dysfunction is challenging and many clinical trials involving patients on dialysis have failed, thus far. Treatment with dialytic techniques - of which maintenance thrice-weekly hemodialysis is most commonly used - is broadly named kidney 'replacement' therapy, a term that casts the perception of a priori abandonment of intrinsic kidney function and subsumes patients into a single, homogeneous group. RECENT FINDINGS: Patients with advanced kidney failure necessitating dialytic therapy may have ongoing endogenous kidney function, and differ in their clinical manifestations and needs. Different terminology, for example, kidney dysfunction requiring dialysis (KDRD) with stages of progressive severity could better capture the range of phenotypes of patients who require kidney 'assistance' therapy. SUMMARY: Classifying patients with KDRD based on objective, quantitative levels of endogenous kidney function, as well as patient-reported symptoms and quality of life, would facilitate hemodialysis prescriptions tailored to level of kidney dysfunction, clinical needs, and personal priorities. Such classification would encourage clinicians to move toward personalized, physiological, and adaptive approach to hemodialysis therapy.

Glypican-3 Differentiates Intraductal Carcinoma and Paget's Disease from Other Types of Breast Cancer.

Background and Objectives: breast cancer remains the most common health burden affecting females worldwide. Despite developments in breast cancer diagnostic approaches and treatment strategies, the clinical management of metastatic breast cancer remains challenging. Thus, there is a need to identify new biomarkers and novel drug targets for breast cancer diagnosis and therapy. Recently, aberrant glypican-3 (GPC3) expression in cancers has gained considerable interest in cancer research. The studies, however, have yielded contradictory results about GPC3 expression in breast cancer. Therefore, the current study aims to analyse GPC3 expression across a large panel of different breast cancer subtypes. Materials and Methods: GPC3 expression was immunohistochemically evaluated in 230 breast cancer patients along with eight normal tissues and its associations to clinical and demographic characteristics, as well as immunohistochemical biomarkers for breast cancer. Moreover, a public database consisting of breast cancer patients' survival data and GPC3 gene expression information was used to assess the prognostic value of GPC3 in the survival of breast cancer patients. Results: GPC3 expression was only characterised in 7.5% of different histological breast cancer subtypes. None of the normal breast tissues displayed GPC3 expression. Interestingly, all cases of Paget's disease, as well as 42.9% of intraductal and 16.7% of mucinous carcinomas were found to have GPC3 expression, where it was able to significantly discriminate Paget's disease and intraductal carcinoma from other breast cancer subtypes. Importantly, GPC3 expression was found more often in tumours that tested positive for the expression of hormone receptors and human epidermal growth factor receptor 2 (HER2), indicating more favourable histological subtypes of breast cancer. Consequently, longer relapse-free survival (RFS) was significantly correlated with higher GPC3 mRNA expression. Conclusions: Our study proposes that GPC3 is a promising breast cancer subtype-specific biomarker. Moreover, GPC3 may have the potential to be a molecular target for the development of new therapeutics for specific subtypes of breast cancer.

Mapping the effect of drugs on ACE2 as a novel target site for COVID-19 therapy.

Angiotensin converting enzyme 2 (ACE2) has potentially conflicting roles in health and disease. COVID-19 coronavirus binds to human cells via ACE2 receptor, which is expressed on almost all body organs. Boosting the ACE2 receptor levels on heart and lung cells may provide more cellular enter to virus thereby worsening the infection. Therefore, among the drug targets, ACE2 is suggested as a vital target of COVID-19 therapy. This hypothesis is based on the protective role of the drugs acting on ACE2. Therefore, this review discusses the impact and challenges of using ACE2 as a target in the current therapy of COVID-19.

Prevalence of erectile dysfunction among Egyptian male patients with type 2 diabetes mellitus.

BACKGROUND AND AIMS: The relation between type 2 diabetes mellitus (T2DM) and erectile dysfunction (ED) has been identified in multiple studies. The aim of this cross-sectional study was to estimate the prevalence and to determine some associated factors of ED among a sample of adult Egyptian male patients with T2DM. METHODS: This cross-sectional study included 150 adult male patients with T2DM (aged 40-60 years) who attended the outpatient clinic of Diabetes in Alexandria Main University hospital. They were evaluated for the presence of ED which was assessed by the validated Arabic-translated five-item version of the International Index of Erectile Function-5 (IIEF-5) questionnaire. Fasting blood glucose (FBG), HbA1c, total serum cholesterol, HDL-C, total serum testosterone (TT) and urinary albumin creatinine ratio (uACR) were measured for all study subjects. RESULTS: The prevalence of ED was 80% among the studied sample. Significant negative correlation was found between IIEF-5 score and age, duration of diabetes, FBG and urinary ACR; while there was a significant positive correlation between IIEF-5 score and serum total testosterone. On performing multiple linear regression analysis for the parameters affecting IIEF-5 questionnaire score, TT, urinary ACR, age and FBG were the independent predictors of ED. CONCLUSION: ED was a common finding in our sample of Egyptian men with T2DM. Poor glycemic control and albuminuria may be considered as independent risk factors for ED.

In Vitro Comparative Study of Emulsified Fat Grafts.

Introduction: Fat grafting is considered one of the most precious armamentarium in the hand of plastic surgeons. The fat grafts consist of 2 components, adipocytes and stromal cells. The adipose tissue is a reserve of stem cells. Aim: The aim of this study was to compare the adipocyte and stem cell viability in both mechanically processed and enzymatically digested fats. Patients and Methods: This in vitro study was conducted using 40 specimens collected from 20 patients who underwent liposuction procedures. Twenty specimens were mechanically processed (group A), whereas the remaining specimens were processed enzymetically (group B). Results: There were no statistically significant differences between fat cell viability between the 2 groups. On the contrary, there was statistically significant increase in stem cells in mechanically processed fat specimens (P = .001). Conclusion: Both the mechanically and chemically processed fat techniques are reliable techniques that provide fat and stem cells. Mechanical processing is easier and provides more stem cells.

Heavy metals-resistant bacteria (HM-RB): Potential bioremediators of heavy metals-stressed Spinacia oleracea plant.

Microorganism technologies can provide a potential alternative to traditional methods of removing heavy metals to conserve agricultural soils. This study aimed to identify and characterize heavy metals-resistant bacteria (HM-RB) isolated from industry-affected soil and their desired impact as bioremediators of heavy metals-stressed spinach plants. Three of 135 isolates were selected based on a high level of resistance to heavy metals. Based on morphological and biochemical characteristics, the selected isolates were identified as Bacillus subtilis subsp. spizizenii DSM 15029 T DSM (MA3), Paenibacillus jamilae DSM 13815 T DSM (LA22), or Pseudomonas aeruginosa DSM 1117 DSM (SN36). Experiments were implemented to investigate the three isolated HM-RB ability on improving attributes of growth, physio-biochemistry, and components of the antioxidant defense system of spinach plant exposed to the stress of cadmium (Cd2+; 2 mM), lead (Pb2+; 2 mM) or 2 mM Cd2++2 mM Pb2+. Compared to control, Cd2+ or Pb2+ stress markedly lowered plant fresh and dry weights, leaf contents of chlorophylls and carotenoids, rates of transpiration (Tr), net photosynthesis (Pn) and stomatal conductance (gs), relative water content (RWC), and membrane stability index (MSI). In contrast, contents of α.tochopherol (α.TOC), ascorbic acid (AsA), glutathione (GSH), proline, soluble sugars, Cd2+, and Pb2+, as well as activities of enzymatic and non-enzymatic antioxidants were markedly elevated. The application of HM-RB promoted the tolerance to heavy metal stress in spinach plants by improving Tr, Pn, gs, RWC, and MSI, while activities of enzymatic and non-enzymatic antioxidants were suppressed. These results reflected positively in promoting plant growth under heavy metal stress. Therefore, the application of HM-RB as potential bioremediators may be a promising strategy for promoting plant growth and productivity under heavy metal stress.

Rapid Room-Temperature Synthesis of Mesoporous TiO2 Sub-Microspheres and Their Enhanced Light Harvesting in Dye-Sensitized Solar Cells.

Submicron sized mesoporous spheres of TiO2 have been a potential alternative to overcome the light scattering limitations of TiO2 nanoparticles in dye-sensitized solar cells (DSSCs). Currently available methods for the growth of mesoporous TiO2 sub-microspheres involve long and relatively high temperature multi-stage protocols. In this work, TiO2 mesoporous sub-microspheres composed of ~5 nm anatase nanocrystallites were successfully synthesized using a rapid one-pot room-temperature CTAB-based solvothermal synthesis. X-Ray Diffraction (XRD) showed that the grown structures have pure anatase phase. Transmission electron microscopy (TEM) revealed that by reducing the surfactant/precursor concentration ratio, the morphology could be tuned from monodispersed nanoparticles into sub-micron sized mesoporous beads with controllable sizes (50-200 nm) and with good monodispersity as well. The growth mechanism is explained in terms of the competition between homogeneous nucleation/growth events versus surface energy induced agglomeration in a non-micelle CTAB-based soft templating environment. Further, dye-sensitized solar cells (DSSCs) were fabricated using the synthesized samples and characterized for their current-voltage characteristics. Interestingly, the DSSC prepared with 200 nm TiO2 sub-microspheres, with reduced surface area, has shown close efficiency (5.65%) to that of DSSC based on monodispersed 20 nm nanoparticles (5.79%). The results show that light scattering caused by the agglomerated sub-micron spheres could compensate for the larger surface areas provided by monodispersed nanoparticles.

Stimulating antioxidant defenses, antioxidant gene expression, and salt tolerance in Pisum sativum seedling by pretreatment using licorice root extract (LRE) as an organic biostimulant.

Plant extracts have recently been used as exogenous adjuvants to strengthen the endogenous plant defense systems when they grow under different environmental stresses, including salinity. The study aimed at determining the effects of seed soaking using licorice root extract (LRE) on photosynthesis and antioxidant defense systems, including transcript levels of enzyme-encoding genes in pea seedling grown under 150 mM NaCl-salinity. Salt stress reduced seedling growth, photosynthesis attributes, and K+ content, and increased oxidative stress (O2•‒ and H2O2, and MDA), Na+, and Cl-, along with an increase in antioxidative defense activities compared to control. However, LRE pretreatment enhanced seedling growth, photosynthetic attributes (chlorophylls, carotenoids, Fv/Fm, Pn, Tr, and gs), ascorbate and glutathione and their redox states, proline, soluble sugars, α-TOC, and enzyme activities compared to stressed control. LRE pretreatment also upregulated transcript levels of CAT-, SOD-, APX-, GR-, DHAR-, and PrxQ-encoding genes in salt-stressed seedlings, decreasing oxidative stress and Na+ and Cl- contents and increasing K+ content and K+/Na+ ratio.

Comparative evaluation of surface roughness of posterior primary zirconia crowns.

AIMS: To compare the surface roughness of four commercially available posterior zirconia crowns on the occlusal surface and occlusal edge (buccal cusps) of first and second primary molars crowns. METHODS: Surface roughness of 40 posterior primary zirconia crowns was measured using a mechanical stylus profilometer. Ten mandibular right molar crowns, consisting of five first primary molar and five second primary molar crowns from four brands-Cheng, Sprig EZCrowns, NuSmile and Kinder Krowns were selected. Mean roughness, Ra and mean roughness depth, Rz was measured for all crowns on two selected surfaces, occlusal surface and buccal cusp tips. Data was evaluated by one way analysis of variance and Tukey's honestly significant difference test at 0.05 level of significance. RESULTS: Statistically significant differences were observed in the mean Ra and Rz values at both selected surfaces among four prefabricated paedodontic zirconia crowns. Kinder Krowns had higher Ra and Rz values compared to Cheng, Sprig EZCrowns and NuSmile. Roughness profile of Kinder Krowns also showed higher vertical scale values co-relating with higher Ra and Rz scores, irrespective of the measurements taken on relatively flat surfaces (occlusal edge) or deeper surface (occlusal pits and fissures). CONCLUSIONS: Mechanically polished posterior primary zirconia crowns had a smoother surface profile than the combined polished-glazed primary zirconia crowns. Cheng Crowns had the lowest mean Ra and Rz values although not statistically significant from Sprig EZCrowns and Nu Smile. Kinder Krowns had the highest mean Ra and Rz scores than other crown groups.

Does the application of silicon and Moringa seed extract reduce heavy metals toxicity in potato tubers treated with phosphate fertilizers?

Two field trials were carried out in two successive agricultural seasons to study the possibility of using silicon (Si) and Moringa seed extract (MSE) for reducing heavy metal contamination resulting from phosphate fertilizers addition to potato tubers. A randomized complete block design experiment was performed using three replicates. Various sources of phosphate fertilizers as ordinary super phosphate and rock phosphate were added at rate of 100 kg P ha-1 prior sowing. Silicon was added as potassium silicate (20% SiO2) at rate of 6 L ha-1, and MSE was also added at rate of 150 L ha-1 in three equal doses with the 2nd, 4th, and 6th irrigations during the last 10 min of drip irrigation. Results indicated that the addition of phosphate fertilizers increased fresh tuber yield, dry weight yield, NPK uptake, catalase, peroxidase, superoxide dismutase, and glutathione reductase of potato either alone or combined with silicon and MSE. The accumulation rate of Cu, Cd, and Ni in potato was higher with the single addition of rock phosphate fertilizer compared with single addition of super phosphate fertilizer. The highest reduction (P < 0.05) in heavy metal accumulation in potato leaves and tubers as well as soil was found with MSE treatment plus super phosphate fertilizer. It is recommended to add MSE at a rate of 150 L ha-1 along with fertilizing the potato crop with ordinary super phosphate fertilizer.

Prevalence and Molecular Characterization of Antimicrobial Resistance in Escherichia coli Isolated from Raw Milk and Raw Milk Cheese in Egypt.

The goal of this study was to examine antimicrobial resistance and characterize the implicated genes in 222 isolates of Escherichia coli from 187 samples of raw milk and the two most popular cheeses in Egypt. E. coli isolates were tested for susceptibility to 12 antimicrobials by a disk diffusion method. Among the 222 E. coli isolates, 66 (29.7%) were resistant to one or more antimicrobials, and half of these resistant isolates showed a multidrug resistance phenotype (resistance to at least three different drug classes). The resistance traits were observed to tetracycline (27.5%), ampicillin (18.9%), streptomycin (18.5%), sulfamethoxazole-trimethoprim (11.3%), cefotaxime (4.5%), kanamycin (4.1%), ceftazidime (3.6%), chloramphenicol (2.3%), nalidixic acid (1.8%), and ciprofloxacin (1.4%). No resistance to fosfomycin and imipenem was observed. Tetracycline resistance genes tetA, tetB, and tetD were detected in 53 isolates, 9 isolates, and 1 isolate, respectively, but tetC was not detected. Aminoglycoside resistance genes strA, strB, aadA, and aphA1 were detected in 41, 41, 11, and 9 isolates, respectively. Sulfonamide resistance genes sul1, sul2, and sul3 were detected in 7, 25, and 3 isolates, respectively. Of 42 ampicillin-resistant isolates, blaTEM, blaCTX-M, and blaSHV were detected in 40, 9, and 3 isolates, respectively, and 10 (23.8%) ampicillin-resistant isolates were found to produce extended-spectrum ß-lactamase. Each bla gene of extended-spectrum ß-lactamase-producing E. coli was further subtyped to be blaCTX-M-15, blaCTX-M-104, blaTEM-1, and blaSHV-12. The class 1 integron was also detected in 28 resistant isolates, and three different patterns were obtained by PCR-restriction fragment length polymorphism. Sequencing analysis of the variable region revealed that four isolates had dfrA12/orfF/aadA2, two had aadA22, and one had dfrA1/aadA1. These data suggest that antimicrobial-resistant E. coli are widely distributed in the milk production and processing environment in Egypt and may play a role in dissemination of antimicrobial resistance to other pathogenic and commensal bacteria.

Toxoplasma genotyping in congenital toxoplasmosis in Upper Egypt: evidence of type I strain.

Toxoplasma gondii has subpopulation structures in different geographical regions caused by less frequent sexual recombination, population sweeps, and biogeography. The majority of strains isolated in North America and Europe fall into one of three clonal lineages, referred to as types I, II, and III. So far, little is known about genetics of Toxoplasma strains in Africa. The present study aimed to determine the genotype of Toxoplasma strains obtained directly from trophoblastic/placental tissues of 29 complicated pregnant women using multilocus nested-PCR-RFLP technique depending on four independent genetic loci (5' SAG2 and 3' SAG2), SAG3, GRA6, and BTUB genes. All samples gave positive amplicons at 5'-3' SAG2 and SAG3 genes. Meanwhile, no amplification products were observed in 12 (41.37%) and 10 (34.48%) samples with GRA6 and BTUB genes, respectively. The restriction pattern revealed the presence of genotype I in all samples, except one sample, which revealed atypical genotype with unusual restriction pattern at 3' SAG2 gene. The negative amplifications in some samples could be due to presence of mutations or polymorphisms in the primer binding sites of these isolates, raising the possibility of mixed or recombinant genotypes. To the best of our knowledge, this is the first time to perform genotype analysis study based on Multiplex nPCR-RFLP technique for genetic characterization of T. gondii in Egypt. Besides, it is the first time to prove that the most prevalent strain of T. gondii, responsible for congenital toxoplasmosis in Upper Egypt, is the highly virulent type I. Atypical genotype was detected as well.

Sex-specific differences in mitochondria biogenesis, morphology, respiratory function, and ROS homeostasis in young mouse heart and brain.

Sex-specific differences in mitochondrial function and free radical homeostasis are reported in the context of aging but not well-established in pathogeneses occurring early in life. Here, we examine if sex disparity in mitochondria function, morphology, and redox status starts early and hence can be implicated in sexual dimorphism in cardiac as well as neurological disorders prevalent at young age. Although mitochondrial activity in the heart did not significantly vary between sexes, female brain exhibited enhanced respiration and higher reserve capacity. This was associated with lower H2O2 production in female cardiac and brain tissues. Using transmission electron microscopy, we found that the number of female cardiac mitochondria is moderately greater (117 ± 3%, P = 0.049, N = 4) than male's, which increased significantly for cortical mitochondria (134 ± 4%, P = 0.001, N = 4). However, male's cardiac mitochondria exhibited fragmented, circular, and smaller mitochondria relative to female's mitochondria, while no morphologic sex-dependent differences were observed in cortical mitochondria. No sex differences were detected in Nox2 and Nox4 proteins or O2-consuming/H2O2-producing activities in brain homogenate or synaptosomes. However, a strong trend of increased EPR-detected NOX superoxide in male synaptosomes hinted at higher superoxide dismutase activity in female brains, which was confirmed by two independent protocols. We also provide direct evidence that respiring mitochondria generally produce an order-of-magnitude lower reactive oxygen species (ROS) proportions than currently estimated. Our results indicate that sex differences in mitochondrial biogenesis, bioenergetics, and morphology may start at young age and that sex-dependent SOD capacity may be responsible for differences in ROS homeostasis in heart and brain.

Comprehensive growth performance, immune function, plasma biochemistry, gene expressions and cell death morphology responses to a daily corticosterone injection course in broiler chickens.

The massive meat production of broiler chickens make them continuously exposed to potential stressors that stimulate releasing of stress-related hormones like corticosterone (CORT) which is responsible for specific pathways in biological mechanisms and physiological activities. Therefore, this research was conducted to evaluate a wide range of responses related to broiler performance, immune function, plasma biochemistry, related gene expressions and cell death morphology during and after a 7-day course of CORT injection. A total number of 200 one-day-old commercial Cobb broiler chicks were used in this study. From 21 to 28 d of age, broilers were randomly assigned to one of 2 groups with 5 replicates of 20 birds each; the first group received a daily intramuscular injection of 5 mg/kg BW corticosterone dissolved in 0.5 ml ethanol:saline solution (CORT group), while the second group received a daily intramuscular injection of 0.5 ml ethanol:saline only (CONT group). Growth performance, including body weight (BW), daily weight gain (DG), feed intake (FI) and feed conversion ratio (FC), were calculated at 0, 3 and 7 d after the start of the CORT injections. At the same times, blood samples were collected in each group for hematological (TWBC's and H/L ratio), T- and B-lymphocytes proliferation and plasma biochemical assays (total protein, TP; free triiodothyronine hormone, fT3; aspartate amino transaminase, AST; and alanine amino transaminase, ALT). The liver, thymus, bursa of Fabricius and spleen were dissected and weighed, and the mRNA expression of insulin-like growth factor 1 gene (IGF-1) in liver and cell-death-program gene (caspase-9) in bursa were analyzed for each group and time; while the apoptotic/necrotic cells were morphologically detected in the spleen. From 28 to 35 d of age, broilers were kept for recovery period without CORT injection and the same sampling and parameters were repeated at the end (at 14 d after initiation of the CORT injection). In general, all parameters of broiler performance were negatively affected by the CORT injection. In addition, CORT treatment decreased the plasma concentration of fT3 and the mRNA expression of hepatic IGF-1. A significant increase in liver weight accompanied by an increase in plasma TP, AST and ALT was observed with CORT treatment, indicating an incidence of liver malfunction by CORT. Moreover, the relative weights of thymus, bursa and spleen decreased by the CORT treatment with low counts of TWBC's and low stimulation of T & B cells while the H/L ratio increased; indicating immunosuppressive effect for CORT treatment. Furthermore, high expression of caspase-9 gene occurred in the bursa of CORT-treated chickens, however, it was associated with a high necrotic vs. low apoptotic cell death pathway in the spleen. Seven days after termination of the CORT treatment in broilers, most of these aspects remained negatively affected by CORT and did not recover to its normal status. The current study provides a comprehensive view of different physiological modulations in broiler chickens by CORT treatment and may set the potential means to mount a successful defense against stress in broilers and other animals as well.

Prevalence and pathogenic potential of Escherichia coli isolates from raw milk and raw milk cheese in Egypt.

The objectives of this study were to investigate prevalence and pathogenic potential of Escherichia coli contaminating raw milk and its products in Egypt. Out of 187 dairy products including 72 raw milk samples, 55 Karish cheese and 60 Ras cheese, 222 E. coli isolates including 111, 89 and 22 were obtained from 55 raw milk samples (76.4%), 41 Karish cheese (74.5%), and 13 Ras cheese (21.7%), respectively. Isolated E. coli strains were examined for 24 representative virulence genes present in diarrheagenic E. coli (DEC) and extraintestinal pathogenic E. coli (ExPEC). Among DEC and ExPEC virulence factors, genes for enteropathogenic E. coli (eaeA, bfpA, EAF), enterohemorrhagic E. coli (stx1, stx2, eaeA), enterotoxigenic E. coli (elt, est), enteroinvasive E. coli (invE), enteroaggregative E. coli (Eagg, astA), diffusely adherent E. coli (daaD), ExPEC (cdt-I to cdt-V, cnf1, cnf2, hlyA) and putative adhesins (efa1, iha, ehaA, saa, and lpfAO113) were screened by colony hybridization assay. Out of 222 E. coli strains, 104 (46.8%) isolated from 69 (36.9%) samples carried one or more virulence genes. The most prevalent gene detected was lpfAO113 (40.5%), followed by ehaA (32.4%,), astA (3.15%,), iha (1.80%), hlyA (1.35%), stx1 (0.90%), stx2 (0.90%), eaeA (0.45%), cdt-III (0.45%) and cnf2 (0.45%). Two strains isolated from Karish cheese harbored 5 virulence genes (stx1, stx2, iha, ehaA, lpfAO113). Stx subtype was determined to be stx1 (not stx1c or stx1d) and stx2d. Indeed, expression of hemolysin A, CDT-III, CNF-II, Stx1 and Stx2d was confirmed by blood agar plate, cytotoxicity assay and Western blotting, respectively. Among the 222 E. coli strains, 54 (48.6%), 38 (42.6%) and 12 (54.7%) isolated from raw milk, Karish cheese and Ras cheese were potentially virulent, respectively. O-genotyping indicated that most of the potentially virulent E. coli isolates did not belong to clinically important O serogroups except O75, O91 and O166, which have been associated with human diseases. Phylogenetic grouping revealed that 150 (67.6%), 67 (30.2%) and 5 (2.30%) strains were clustered into A, B1 and D groups, respectively, which are considered to be associated with intestinal infection, indicating that these E. coli strains might have a potential to cause gastroenteritis. To the best of our knowledge, this is the first comprehensive study regarding prevalence and pathogenic potential of E. coli in dairy products in Egypt. Raw milk, Karish cheese and Ras cheese in Egypt are highly contaminated with E. coli including potentially pathogenic strains, which may impose a public health threat.