Evaluation of Circular RNA SMARCA5 as a Novel Biomarker for Hepatocellular Carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is the fourth most prevalent type of cancer in Egypt and the sixth globally. Most patients with HCC are typically diagnosed during the advanced stages of the disease due to the absence of biomarkers for early detection. Consequently, these patients miss the optimal timeframe for receiving therapy. OBJECTIVE: we aimed to assess the circular RNA SMARCA5 level and SMARCA5 mRNA gene expression as a potential biomarker for early detection of HCC. METHODS: The present study utilized a case-control design comprising 159 participants. Participants were selected from both inpatient and outpatient hepatology and gastroenterology clinics at the National Liver Institute Hospital, Menoufia University. They were evenly distributed among three groups: Group I: 53 control subjects, Group II: 53 HCV cirrhotic patients, and Group III: 53 HCC patients. Tumor staging was done using BCLC staging system. Each patient underwent a thorough clinical examination, radiological examination, complete history taking, and serum Alpha-fetoprotein (AFP) assessment and detection of circular RNASMARCA5 and SMARCA5mRNA gene sutilizing quantitative real-time polymerase chain reaction. RESULTS: Statistically substantial differences were observed in the examined groups in terms of AFP, SMARCA5, and CircSMARCA5 (P-value = 0.001, 0.001 & 0.001). CircSMARCA5 and SMARCA5mRNA were markedly down regulated in the HCC group compared to HCV cirrhotic patients and controls. ROC analysis for early HCC diagnosis demonstrated that the CircSMARCA5 area under the curve (AUC) at cut-off point 4.55 yielded a specificity of 83.8% and sensitivity of 91.7%. The AUC for AFP at a cut-off point of 515ng/ml yielded a specificity of 89.2% and a sensitivity of 91.3%. CONCLUSION: CircSMARCA5 has the potential to be a more sensitive predictor of HCC disease compared to AFP.

Assessing agronomic performance, chocolate spot resistance, and heat tolerance for diverse Vicia faba genotypes under varying environmental conditions.

Chocolate spot and heat stress devastatingly impact the production of faba bean, particularly under prevailing climatic changes and rising drastic environmental conditions. Hence, the adaptability of faba bean performance is a decisive objective of plant breeders to ensure its sustainable production. The present study aimed to evaluate the agronomic performance and stability of diverse eleven faba bean genotypes for yield characters, chocolate spot, and heat stress in eight different growing environments. The faba bean genotypes were evaluated at two sowing dates in two different locations during two growing seasons. The evaluated eleven faba bean genotypes were sown timely in autumn (25 October) and late sowing in early winter (25 November) in Bilbeis and Elkhatara during 2020 and 2021 growing seasons. The results exhibited substantial differences among the evaluated sowing dates, locations, and faba bean genotypes for all studied characters. The genotypes Sakha-3, Nubaria-3, Nubaria-5, Misr-3, and Wadi-1 were able to produce acceptable yield and quality characters under timely sowing in autumn and late sowing in early winter in all tested environments. Moreover, the genotypes Nubaria-3, Nubaria-4, Nubaria-5, Sakha-4, Giza-3, and Triple White exhibited better resistance to chocolate spot. The assessed faba bean genotypes were evaluated under late sowing to expose the plants to high temperature stress at flowering and throughout the anthesis and seed-filling stages. The genotypes Nubaria-5, Nubaria-3, Nubaria-4, Sakha-3, Sakha-4, Wadi-1, and Misr-3 possessed tolerance to heat stress more than the other genotypes. Different statistical methods were applied to study the stability of assessed genotypes such as joint regression, Additive Main Effect and Multiplicative Interaction (AMMI) analysis, AMMI stability value, Wricke's and Ecovalence values. The estimated stability parameters were consistent in depicting the stability of the assessed faba bean genotypes. The findings revealed that Sakha-1, Misr-3, Nubaria-4, and Nubaria-5 demonstrated stable and desirable performance across all tested environments. The heatmap was employed to classify the assessed faba bean genotypes into different groups based on agronomic performance, chocolate spot resistance and heat stress tolerance. Nubaria-3, Nubaria-4, Nubaria-5, and Misr-3 had the best performance for agronomic performance, chocolate spot resistance, and heat stress tolerance. The obtained results provide evidence of employing promising faba bean genotypes for improving the stability of agronomic performance, chocolate spot resistance, and heat stress tolerance in breeding programs principally under unprecedented climate fluctuations.

The use of lung ultrasound in evaluation of extravascular lung water in hemodialysis patients: Systematic review and meta-analysis.

RATIONALE AND OBJECTIVES: Determining dry weight is crucial for optimizing hemodialysis, influencing efficacy, cardiovascular outcomes, and overall survival. Traditional clinical assessment methods for dry weight, relying on factors such as blood pressure and edema, frequently lack reliability. Lung ultrasound stands out as a promising tool for assessing volume status, given its non-invasiveness and reproducibility. This study aims to explore the role of Lung ultrasound in evaluating the impact of hemodialysis and ultrafiltration on extravascular lung water, with a specific focus on changes in B-lines post-hemodialysis compared to pre-hemodialysis. MATERIALS AND METHODS: The research encompassed searches across PubMed, WOS, and Scopus databases for studies related to lung ultrasound and hemodialysis. A meta-analysis was then performed to determine the mean differences in various parameters before compared to after, hemodialysis, including the number of B-lines, indexed end-inspiratory and end-expiratory inferior vena cava diameters, inferior vena cava collapsibility index, weight, blood pressure, and serum levels of NT-pro-BNP. RESULTS: Our meta-analysis, included 33 studies with 2301 hemodialysis patients, revealed a significant decrease in the number of B-lines post-hemodialysis (mean difference = 8.30, 95% CI [3.55 to 13.05]). Furthermore, there was a noteworthy reduction in inspiratory and expiratory inferior vena cava diameters post-hemodialysis (mean difference = 2.32, 95% CI [0.31 to 4.33]; mean difference = 4.05, 95% CI [2.44 to 5.65], respectively). Additionally, a significant positive correlation was observed between B-lines and the maximum inferior vena cava diameter both pre- and post-hemodialysis (correlation coefficient = 0.39; correlation coefficient = 0.32, respectively). CONCLUSION: These findings indicate the effectiveness of lung ultrasound in detection of volume overload and assessment of response to ultrafiltration in hemodialysis patients.

Novel VEGFR2 inhibitors with thiazoloquinoxaline scaffold targeting hepatocellular carcinoma with lower cardiotoxic impact.

Hepatocellular carcinoma (HCC) is a fatal tumor which is usually diagnosed at advanced stage. Molecular targeted drugs were used recently to treat HCC, however, due to serious side effects, mainly cardiotoxicity and emergence of resistance, there is demanding to explore new chemotherapeutics. 10 novel thiazoloquinoxaline derivatives coupled with different sulfonamide moieties 4(a-j) were designed and synthesized fulfilling pharmacophoric features of VEGFR-2 inhibition. Structures of all new compounds were verified via spectral and microanalytical data. After carrying in-vitro VEGFR-2 assay for compounds 4(a-j); sulfapyridine and sulfamethoxazole derivatives 4d and 4f showed potential inhibitory effect [61.04 and 83.35 nM], respectively, comparable to standard sorafenib [51.41 nM]. Both were then further evaluated for their cytocidal activity against HepG2 cell-line and against myocardium cells using H9C2 cell-line. As a result, only sulfapyridine derivative 4d exhibited a significant inhibition of HepG2 cells viability [IC50 = 4.31 µM]. Furthermore, it showed relatively lower cytotoxic impact against normal H9C2 myocardium cells [IC50, 33.47 µM] compared to that of sorafenib [IC50, 98.07 µM]. In-vivo study was carried out to determine myocardium safety of compound 4d on irradiated mice (8 Gy). In-vivo results of sulfapyridine derivative 4d showed normal cardiac enzyme function (CK) and serum catalase activity with significant reductions in LDH, cardiac TNF-α and caspase-9 levels, alongside with its efficacy in suppressing the expression of hepatic VEGF. In conclusion, sulfapyridine derivative 4d could be considered a promising candidate as VEGFR-2 inhibitor with less myocardium side effect.

Response of Diverse Peanut Cultivars to Nano and Conventional Calcium Forms under Alkaline Sandy Soil.

Calcium is one of the most limiting factors for the growth and reproduction of peanut, which ultimately affects pod and seed yields. A two-year field experiment was carried out to assess the impact of five calcium applications, including nano-calcium and conventional forms, on growth, leaf nutrient content, yield traits, and quality parameters of three diverse peanut cultivars (Ismailia-1, Giza-5, and Giza-6). The applied calcium applications were calcium sulfate, which is recommended for commercial peanut cultivation and commonly referred to as gypsum (coded as Ca-1), calcium nitrate (Ca-2), nano-calcium nitrate (Ca-3), 50% calcium nitrate + 50% nano-calcium (Ca-4), and 50% calcium sulfate + 50% nano-calcium (Ca-5). Calcium sulfate (gypsum, Ca-1) was soil-supplied during the seedbed preparation as recommended, while the other calcium applications (Ca-2, Ca-3, Ca-4, and Ca-5) were exogenously sprayed three times at 30, 45, and 60 days after sowing. The soil of the experimental site was alkaline, with a high pH of 8.6. The results revealed significant differences among cultivars, calcium applications, and their interactions. The soil-supplied gypsum Ca-1 displayed lower agronomic performance on all recorded growth, leaf nutrient content, yield traits, and quality parameters. On the other hand, the foliar-supplied calcium, particularly Ca-4 and Ca-5, displayed superior effects compared to the other simple calcium forms. Ca-4 and Ca-5 produced significantly higher seed yield (3.58 and 3.38 t/ha) than the simple recommended form (Ca-1, 2.34 t/ha). This could be due to the difficulty of calcium uptake from soil-supplied calcium under high soil pH compared to the exogenously sprayed nano-calcium form. Moreover, the superior performance of Ca-4 and Ca-5 could be caused by the mixture of fertilizers from the synergistic effect of calcium and nitrate or sulfate. Furthermore, the effect of nitrate was applied in nano form in the Ca4 and Ca-5 treatments, which contributed to improving nutrient uptake efficiency and plant growth compared to the other treatments. The peanut cultivar Giza-6 showed superiority for most measured traits over the other two cultivars. The interaction effect between the assessed cultivars and calcium applications was significant for various traits. The cultivar Giza-6 showed a significant advantage for most measured traits with the mixture of 50% calcium nitrate + 50% nano-calcium (Ca-4). Conclusively, the results pointed out the advantage of the exogenously sprayed nano-calcium form combined with calcium nitrate or calcium sulfate for promoting growth, leaf nutrient content, yield, and quality traits of peanut, particularly with high-yielding cultivars under sandy soil with high pH.

Cause-specific mortality among patients with renal cell carcinoma in the United States from 2000 to 2018.

OBJECTIVES: There has been little focus on the non-cancer causes of death in patients with renal cell carcinoma (RCC). Therefore, we aimed to assess the frequency and risk of different causes of death, stratified by tumor stage, and demographics, after a diagnosis of RCC in the United States. MATERIALS AND METHODS: Data on eligible patients with RCC from January 1, 2000, to December 31, 2018, in the United States were retrieved from the Surveillance, Epidemiology, and End Results (SEER) database. Standardized mortality ratios for causes of death were calculated using the SEER*Stat software 8.3.9.2 for the overall population and stratified subgroups. RESULTS: A total of 165,969 patients with RCC were included and 60,290 (36.3%) died during follow-up. The majority of deaths were due to kidney cancer (51.3%) but a significant proportion was non-cancer causes (37.6%). The proportion of deaths attributed to RCC decreased with increasing follow-up with non-cancer causes becoming dominant after the fifth year following RCC diagnosis. Overall, cardiovascular diseases and cerebrovascular diseases were the most common non-RCC-related causes of death. AJCC stage I and localized RCC had the most deaths attributed to non-cancerous causes (66.2% and 61.2%, respectively) while AJCC stage IV and distant RCC had the most deaths due to RCC (86.2% and 86.5%, respectively). CONCLUSION: A large proportion of RCC patients die of non-cancerous causes especially early-stage patients and advanced-stage patients who survive >5 years. Coordination of multidisciplinary care with relevant specialists depending on the stage of the disease is needed to better prevent death overtime from non-cancer causes.

Prediction of heavy metal biosorption mechanism through studying isotherm kinetic equations.

The kinetic constants for free and immobilized cells were determined by measuring reaction rates at different metal concentrations at the optimum reaction conditions. (Kmax and Vmax) were calculated from the slope and intercept of the straight lines. The pseudo-second-order rate constants are derived based on the sorption capacity of the solid phase, where K2 is the rate constant for the pseudo-second-order model. Determined experimentally by plotting t/q against t. The mean free vitality of adsorption (E) was figured as 2.62 kJ mol-1 and the extent of E communicated gives data on the adsorption mechanism. An E value ranging from 1 to 8 indicates physisorption and 8-16 kJ mol-1 predicts ion exchange. Thus, the evaluated value of 2.62 kJ mol-1 predicts the phenomenon of physisorption, which suggests that metal ions were favorably adsorbed by this biosorbent in a multi-layer fashion. The overall result suggested that 98.2% of U (VI) by biosorption of U in the mechanism of adsorption will include chemisorption mechanistic pathway: Langmuir, Freundlich, equations and the values of Kf 5.791 where KL 3.9 were determined from the linear plot of log qe vs. log Ce at 30 °C, indicating that metal ions were favorably adsorbed by this biosorbent in a multi-layer fashion and instrumentation of beads characterizing novel Binding sites using FTIR & SEM beside change in peaks position which assigned for its groups confirm biosorption of metal.

Aspergillus Niger thermostable Cytosine deaminase-dextran conjugates with enhanced structure stability, proteolytic resistance, and Antiproliferative activity.

Cytosine deaminase (CDA) is a prodrug mediating enzyme converting 5-flurocytosine into 5-flurouracil with profound broad-range anticancer activity towards various cell lines. Availability, molecular stability, and catalytic efficiency are the main limiting factors halting the clinical applications of this enzyme on prodrug and gene therapies, thus, screening for CDA with unique biochemical and catalytic properties was the objective. Thermotolerant/ thermophilic fungi could be a distinctive repertoire for enzymes with affordable stability and catalytic efficiency. Among the recovered thermotolerant isolates, Aspergillus niger with optimal growth at 45 °C had the highest CDA productivity. The enzyme was purified, with purification 15.4 folds, molecular mass 48 kDa and 98 kDa, under denaturing and native PAGE, respectively. The purified CDA was covalently conjugated with dextran with the highest immobilization yield of 75%. The free and CDA-dextran conjugates have the same optimum pH 7.4, reaction temperature 37 °C, and pI 4.5, and similar response to the inhibitors and amino acids suicide analogues, ensuring the lack of effect of dextran conjugation on the CDA conformational structure. CDA-Dextran conjugates had more resistance to proteolysis in response to proteinase K and trypsin by 2.9 and 1.5 folds, respectively. CDA-Dextran conjugates displayed a dramatic structural and thermal stability than the free enzyme, authenticating the acquired structural and catalytic stability upon dextran conjugation. The thermal stability of CDA was increased by about 1.5 folds, upon dextran conjugation, as revealed from the half-life time (T1/2). The affinity of CDA-conjugates (Km 0.15 mM) and free CDA (Km 0.22 mM) to deaminate 5-fluorocytosine was increased by 1.5 folds. Upon dextran conjugation, the antiproliferative activity of the CDA towards the different cell lines "MDA-MB, HepG-2, and PC-3" was significantly increased by mediating the prodrug 5-FC. The CDA-dextran conjugates strongly reduce the tumor size and weight of the Ehrlich cells (EAC), dramatically increase the titers of Caspase-independent apoptotic markers PARP-1 and AIF, with no cellular cytotoxic activity, as revealed from the hematological and biochemical parameters.

Computational, in vitro and radiation-based in vivo studies on acetamide quinazolinone derivatives as new proposed purine nucleoside phosphorylase inhibitors for breast cancer.

The present work describes a quinazolinone-based lead optimization for the development of novel purine nucleoside phosphorylase (PNP) inhibitors with quinazolinone scaffold. Nineteen compounds were proposed and docked against PNP, the best 14 compounds with highest docking and affinity scores and low RMSD values were synthesized. Synthesis of new quinazolinone derivatives with variable acetamide substituents on two positions on quinazoline ring was performed. The structures assigned to the products were concordant with the microanalytical and spectral data. In vitro cytotoxicity on human breast cancer cell line (MCF7) was performed and identified compound 6g as the most potent with IC50 (0.99 ± 0.11 µM) which was further tested against five different breast cancer cell lines in addition to normal breast cell to determine the selectivity. Compound 6g was subjected to molecular dynamic simulation study, radiolabelling and biodistribution study to investigate its stability and selectivity toward breast cancers. The in vitro PNP inhibition results were aligned with the in silico, cytotoxicity, and biodistribution results where 6g showed the most potent PNP inhibitory activity with IC50 (0.159 ± 0.007 µM) when compared to Peldesine (BCX-34) IC50 (0.041 ± 0.002 µM).

AI-based adaptive personalized content presentation and exercises navigation for an effective and engaging E-learning platform.

Effective and engaging E-learning becomes necessary in unusual conditions such as COVID-19 pandemic, especially for the early stages of K-12 education. This paper proposes an adaptive personalized E-learning platform with a novel combination of Visual/Aural/Read, Write/Kinesthetic (VARK) presentation or gamification and exercises difficulty scaffolding through skipping/hiding/ reattempting. Cognitive, behavior and affective adaptation means are included in developing a dynamic learner model, which detects and corrects each student's learning style and cognitive level. As adaptation targets, the platform provides adaptive content presentation in two groups (VARK and gamification), adaptive exercises navigation and adaptive feedback. To achieve its goal, the platform utilizes a Deep Q-Network Reinforcement Learning (DQN-RL) and an online rule-based decision making implementation. The platform interfaces front-end dedicated website and back-end adaptation algorithms. An improvement in learning effectiveness is achieved comparing the post-test to the pre-test in a pilot experiment for grade 3 mathematics curriculum. Both groups witnessed academic performance and satisfaction level improvements, most importantly, for the students who started the experiment with a relatively low performance. VARK group witnessed a slightly more improvement and higher satisfaction level, since interactive activities and games in the kinesthetic presentation can provide engagement, while keeping other presentation styles available, when needed.

Serum Zinc Levels in Chronic Kidney Disease Patients, Hemodialysis Patients, and Healthy Controls: Systematic Review and Meta-Analysis.

OBJECTIVES: Patients with chronic kidney disease (CKD) are susceptible to changes in zinc homeostasis through anorexia and dietary restrictions, as well as hemodialysis (HD). Changes in zinc homeostasis might predispose CKD and HD patients to specific adverse effects, including erythropoietin-resistant anemia, oxidative stress, atherosclerosis, and cardiovascular disease. Because serum zinc levels are rarely measured in CKD and HD patients, zinc supplementations do not represent a routine therapy for CKD and dialysis patients. Therefore, in this meta-analysis, we aimed to assess serum zinc levels in CKD and HD patients compared with healthy controls (HC). In addition, we investigated whether HD affects serum zinc levels by comparing serum zinc levels in HD versus CKD patients and comparing serum zinc pre- versus post-HD. DESIGN AND METHODS: A comprehensive search of databases was conducted to identify either observational studies or randomized trials that assessed serum zinc levels in either CKD and/or HD patients in comparison to HC. We conducted a random-effects meta-analysis. RESULTS: Our meta-analysis included 42 studies with a total of 4,161 participants, of whom 460 were CKD patients, 2,047 were HD patients, and 1,654 were HCs. Both CKD and HD patients showed lower serum zinc levels compared with HC (mean difference = -22.86 µg/dL, 95% CI -33.25 to -12.46; mean difference = -13.64 µg/dL, 95% CI -21.47 to -53.80, respectively). CKD and HD patients showed no significant difference in serum zinc levels (mean difference = 15.39, 95% CI -8.91 to 39.68). Pre-HD serum zinc levels were significantly lower than those post-HD (mean difference = -7.51 µg/dL, 95% CI -14.24 to -0.78). CONCLUSION: In the current study, the serum zinc levels were lower in CKD and HD patients compared to HCs and appears to be more common than reported in daily clinical practice. It may be beneficial to assess serum zinc levels in CKD and HD patients. More research on zinc in kidney disease is encouraged.

Revisiting maintenance immunosuppression in patients with renal transplant failure: early weaning of immunosuppression versus prolonged maintenance-systematic review and meta-analysis.

INTRODUCTION: Prolonged immunosuppression after dialysis start has been assumed to reduce sensitization, need for graft nephrectomy, and to favor re-transplantation. In contrast, immunosuppression is considered to increase the risk of mortality, infection, and malignancy. We aimed to assess the evidence regarding superiority of early or late withdrawal of maintenance immunosuppression post renal transplant failure. METHODS: A literature search of the PubMed, WOS, Ovid, and Scopus databases was conducted. Combined relative risks, (RRs), mean differences, and 95% confidence intervals (CIs) were calculated by using a random-effect model. RESULTS: Ten studies involving 1187 patients with kidney transplant failure were included. No difference could be detected between patients with early withdrawal of  immunosuppressive drugs (≤ 3 months) or prolonged immunosuppressive treatment (> 3 months) regarding mortality (95% CI 0.91-2.28), panel reactive antibodies (PRAs) (95% CI - 0.75-30.10), re-transplantation rate (95% CI 0.55-1.35), infectious episodes (95% CI 0.67, 1.17), cancer (95% CI 0.26-1.54), and graft nephrectomy (95% CI 0.82-1.63). Similarly, no difference was found between immunosuppressive drug withdrawal over < 6 or ≥ 6 months regarding mortality (95% CI 0.16, 2.89), re-transplantation rate (95% CI 0.85-1.55), cancer (95% CI 0.37-1.63), and allograft nephrectomy (95% CI 0.87-4.33). CONCLUSION: Prolonged maintenance immunosuppression post kidney transplant failure is not associated with increased risk of mortality, infection, or malignancy, or reduced risk of sensitization or allograft nephrectomy compared with early withdrawal.

Relation of Liver Siderosis to Liver Fibrosis in Hemodialysis Patients With Severe Hyperferritinemia Secondary to High Doses of Intravenous Iron Supplementation.

OBJECTIVE: Aggressive iron substitution in hemodialysis (HD) patients leads to iron overload. The association between liver siderosis and fibrosis is still debatable. We studied the association of liver siderosis with liver fibrosis in HD patients. Furthermore, we studied the performance of liver stiffness measurements (LSMs) in identifying advanced liver fibrosis. We investigated the performance of biochemical indicators of iron status in identifying advanced liver fibrosis. METHODS: Fifty-five HD patients (average HD duration 6 ± 2 years) with hyperferritinemia secondary to intravenous iron supplementation (weakly iron dose 252.7 ± 63 mg; median blood transfusions 3 [2-5]) were recruited. The liver fibrosis grade was determined with Fibroscan, aminotransferase-to-platelet ratio index (APRI), and Fib-4 index. Liver iron concentration (LIC) was estimated with magnetic resonance imaging (MRI). Iron parameters and liver function biochemical indicators were also assessed. RESULTS: The median serum ferritin and transferrin saturation (TSAT) were 3531 µg/L and 77%, respectively. 34.5%, 20%, and 45.5% of the patients showed mild, moderate, or severe liver siderosis, respectively. All patients with severe liver siderosis showed advanced liver fibrosis. Patients with severe liver siderosis and advanced liver stiffness showed higher serum iron, TSAT, aspartate aminotransferase (AST), alanine aminotransferase (ALT), serum bilirubin, APRI, and Fib-4 index scores than those with mild liver siderosis. Serum iron and TSAT showed good utility in identifying advanced liver fibrosis determined with Fibroscan, APRI, and Fib-4 index. Liver stiffness exhibited good utility in identifying advanced liver fibrosis diagnosed with APRI and Fib-4 index. CONCLUSIONS: High weekly intravenous iron dose associated with severe hyperferritinemia, high serum iron, and TSAT might lead to severe liver siderosis and concomitant liver fibrosis in HD patients. Serum iron, TSAT, Fibroscan, Fib-4, and APRI scores might offer noninvasive tools for identifying advanced liver fibrosis in those patients.

Cholesterol-lowering effects and safety assessment of Lactobacillus spp. in vivo and in vitro testing for human use as probiotic from the dairy product in Egypt.

BACKGROUND: The toxicity profile of lactobacilli may be strain dependent, so it should be considered for safe utilization of probiotics. Further, in vivo studies are necessary to evaluate their safety. RESULT: The ability of various probiotic strains to hydrolyze bile salts has been confirmed without noticeable hemolytic activity. Results revealed the presence of α-glucosidase, ß-glucosidase, α-galactosidase, and ß-galactosidase activity in all investigated isolates, while none of the isolates produced the carcinogenic enzyme ß-glucuronidase. The probiotic strains exhibited remarkable cholesterol-lowering impact. Also, we found no evidence of chronic toxicity under the experimental conditions based on gross pathological examination of the viscera and study of the spleen and liver weight ratios. These findings indicated that the investigated strains, either alone or combined with their metabolites, had no obvious adverse effect on the mice's general health status. CONCLUSION: There is prove that the investigated probiotic strains are safe to be utilized for enhancing of the growth performance and are free of adverse side effects.

Versatile Roles of Microbes and Small RNAs in Rice and Planthopper Interactions.

Planthopper infestation in rice causes direct and indirect damage through feeding and viral transmission. Host microbes and small RNAs (sRNAs) play essential roles in regulating biological processes, such as metabolism, development, immunity, and stress responses in eukaryotic organisms, including plants and insects. Recently, advanced metagenomic approaches have facilitated investigations on microbial diversity and its function in insects and plants, highlighting the significance of microbiota in sustaining host life and regulating their interactions with the environment. Recent research has also suggested significant roles for sRNA-regulated genes during rice-planthopper interactions. The response and behavior of the rice plant to planthopper feeding are determined by changes in the host transcriptome, which might be regulated by sRNAs. In addition, the roles of microbial symbionts and sRNAs in the host response to viral infection are complex and involve defense-related changes in the host transcriptomic profile. This review reviews the structure and potential functions of microbes and sRNAs in rice and the associated planthopper species. In addition, the involvement of the microbiota and sRNAs in the rice-planthopper-virus interactions during planthopper infestation and viral infection are discussed.

Microbial cytosine deaminase is a programmable anticancer prodrug mediating enzyme: antibody, and gene directed enzyme prodrug therapy.

Cytosine deaminase (CDA) is a non-mammalian enzyme with powerful activity in mediating the prodrug 5-fluorcytosine (5-FC) into toxic drug 5-fluorouracil (5-FU), as an alternative directed approach for the traditional chemotherapies and radiotherapies of cancer. This enzyme has been frequently reported and characterized from various microorganisms. The therapeutic strategy of 5-FC-CDA involves the administration of CDA followed by the prodrug 5-FC injection to generate cytotoxic 5-FU. The antiproliferative activity of CDA-5-FC elaborates from the higher activity of uracil pathway in tumor cells than normal ones. The main challenge of the therapeutic drug 5-FU are the short half-life, lack of selectivity and emergence of the drug resistance, consistently to the other chemotherapies. So, mediating the 5-FU to the tumor cells by CDA is one of the most feasible approaches to direct the drug to the tumor cells, reducing its toxic effects and improving their pharmacokinetic properties. Nevertheless, the catalytic efficiency, stability, antigenicity and targetability of CDA-5-FC, are the major challenges that limit the clinical application of this approach. Thus, exploring the biochemical properties of CDA from various microorganisms, as well as the approaches for localizing the system of CDA-5-FC to the tumor cells via the antibody directed enzyme prodrug therapy (ADEPT) and gene directed prodrug therapy (GDEPT) were the objectives of this review. Finally, the perspectives for increasing the therapeutic efficacy, and targetability of the CDA-5-FC system were described.

CD4+CD25+CD127-Foxp3+ and CD8+CD28- Tregs in Renal Transplant Recipients: Phenotypic Patterns, Association With Immunosuppressive Drugs, and Interaction With Effector CD8+ T Cells and CD19+IL-10+ Bregs.

Introduction: Gaps still exist regarding knowledge on regulatory cells in transplant recipients. We studied the phenotypic patterns of CD4+, CD8+CD28- Tregs, and CD19+IL-10+ Bregs in the blood of healthy controls (HC), end-stage kidney disease patients (ESKD), early and late stable renal transplant recipients (Tx), and transplant recipients with steroid-treated acute cellular rejection 1 week-3 months after successful treatment. We also investigated the relationship between immunosuppressive drugs and the aforementioned regulatory cells in transplant recipients. Methods: We recruited 32 HC, 83 ESKD, 51 early Tx, 95 late Tx, and 9 transplant patients with a recent steroid-treated acute cellular rejection. Besides CD19+IL-10+ Bregs, we analyzed absolute and relative frequencies of CD4+CD25+CD127-Foxp3+ Tregs and CD8+CD28- Tregs and their expression of IL-10, TGF-ß, IFN-g, and Helios. Results: We found a negative correlation between absolute CD4+CD25+CD127-Foxp3+ Treg and relative CD19+IL-10+ Breg frequencies in early Tx recipients (r=-0.433, p=0.015, n=31). In that group, absolute CD4+CD25+CD127-Foxp3+ Tregs were negatively associated with steroid dose and tacrolimus trough levels (r=-0.377, p = 0.021, n=37; r=-0.43, p=0.033, n=25, respectively), opposite to IL-10+ Bregs, whose frequency apparently was not negatively affected by potent immunosuppression early posttransplant. We found also lower CD4+CD25+CD127-Foxp3+ Tregs in patients treated with basiliximab or rATG as compared with ESKD patients (p=0.001 and p <0.001, respectively). No difference in absolute IL-10+ Bregs could be detected among these 3 patient groups. Early Tx recipients showed lower CD4+CD25+CD127-Foxp3+ Tregs within 3 months of antibody induction than after 3 months (p = 0.034), whereas IL-10+ Bregs showed higher relative counts during the first 3 months post antibody induction than after 3 months (p = 0.022). Our findings suggest that IL-10+ Bregs decrease with time posttransplantation independent of the effect of antibody induction and dose of other immunosuppressive drugs. Conclusion: These findings suggest that CD19+IL-10+ Bregs and CD4+CD25+CD127-Foxp3+ Tregs behave in opposite ways during the early posttransplant period, possibly due to a predominant negative impact of high doses of immunosuppressants on Tregs. CD19+IL-10+Bregs do not seem to be suppressed by antibody induction and early potent immunosuppression with chemical drugs.

Higher CD19+CD25+ Bregs are independently associated with better graft function in renal transplant recipients.

BACKGROUND: The Identification of B cell subsets with regulatory functions might open the way to new therapeutic strategies in the field of transplantation, which aim to reduce the dose of immunosuppressive drugs and prolong the graft survival. CD25 was proposed as a marker of a B-cell subset with an immunosuppressive action termed Bregs. The effect of CD19 + CD25 + Bregs on graft function in renal transplant recipients has not yet been elucidated. We investigated a potential impact of CD19 + CD25 + Bregs on renal graft function as well as a possible interaction of CD19 + CD25 + Bregs with peripheral Tregs in healthy controls, end-stage kidney disease patients (ESKD), and renal transplant recipients. Moreover, we aimed to investigate the association of CD19 + CD25 + Bregs with serum IL-10, TGF-ß1, and IFN-γ in the same study groups. METHOD: Thirty-one healthy controls, ninety renal transplant recipients, and eighteen ESKD patients were enrolled. We evaluated the CD19 + CD25 + Bregs and Treg absolute counts. Next, we investigated CD19 + CD25 + Bregs as predictors of good graft function in multiple regression and ROC analyses. Finally, we evaluated the association between CD19 + CD25+ Bregs and serum IL-10, TGF-ß, and IFN-γ. RESULTS: ESKD patients and renal transplant recipients showed lower counts of CD19 + CD25+ Bregs compared to healthy controls (p < 0.001). Higher CD19 + CD25+ Breg counts were independently associated with a better GFR in renal transplant recipients (unstandardized B coefficient = 9, p = 0.02). In these patients, higher CD19 + CD25+ Bregs were independently associated with higher Treg counts (unstandardized B = 2.8, p = 0.004). In ROC analysis, cut-offs for CD19 + CD25 + Breg counts and serum TGF-ß1 of 0.12 cell/µl and 19,635.4 pg/ml, respectively, were shown to provide a good sensitivity and specificity in identifying GFR ≥ 30 ml/min (AUC = 0.67, sensitivity 77%, specificity 43%; AUC = 0.65, sensitivity 81%, specificity 50%, respectively). Finally, a significant positive association between CD19 + CD25+ Bregs and TGF-ß1 was shown in renal transplant recipients (r = 0.255, p = 0.015). CONCLUSIONS: Our findings indicate that higher counts of CD19 + CD25+ Bregs are independently associated with better renal function and higher absolute Treg counts in renal transplant recipients.

The Importance of Nitric Oxide as the Molecular Basis of the Hydrogen Gas Fumigation-Induced Alleviation of Cd Stress on Ganoderma lucidum.

Whether or not hydrogen gas (H2) can reduce cadmium (Cd) toxicity in Ganoderma lucidum has remained largely unknown. Here, we report that Cd-induced growth inhibition in G. lucidum was significantly alleviated by H2 fumigation or hydrogen-rich water (HRW), evaluated by lower oxidative damage and Cd accumulation. Moreover, the amelioration effects of H2 fumigation were better than of HRW in an optimum concentration of H2 under our experimental conditions. Further results showed that H2-alleviated growth inhibition in G. lucidum was accompanied by increased nitric oxide (NO) level and nitrate reductase (NR) activity under Cd stress. On the other hand, the mitigation effects were reversed after removing endogenous NO with its scavenger cPTIO or inhibiting H2-induced NR activity with sodium tungstate. The role of NO in H2-alleviated growth inhibition under Cd stress was proved to be achieved through a restoration of redox balance, an increase in cysteine and proline contents, and a reduction in Cd accumulation. In summary, these results clearly revealed that NR-dependent NO might be involved in the H2-alleviated Cd toxicity in G. lucidum through rebuilding redox homeostasis, increasing cysteine and proline levels, and reducing Cd accumulation. These findings may open a new window for H2 application in Cd-stressed economically important fungi.

WITHDRAWN:Randomized controlled trial of effect of N-acetylcysteine as an antioxidant on iron overload in children with thalassemia major.

Ahead of Print article withdrawn by publisher.