Cla4A, a Novel Regulator of Gene Expression Networks Required for Asexual and Insect-Pathogenic Lifecycles of Beauveria bassiana.

Cla4, an orthologous p21-activated kinase crucial for non-entomopathogenic fungal lifestyles, has two paralogs (Cla4A/B) functionally unknown in hypocrealean entomopathogens. Here, we report a regulatory role of Cla4A in gene expression networks of Beauveria bassiana required for asexual and entomopathogenic lifecycles while Cla4B is functionally redundant. The deletion of cla4A resulted in severe growth defects, reduced stress tolerance, delayed conidiation, altered conidiation mode, impaired conidial quality, and abolished pathogenicity through cuticular penetration, contrasting with no phenotype affected by cla4B deletion. In ∆cla4A, 5288 dysregulated genes were associated with phenotypic defects, which were restored by targeted gene complementation. Among those, 3699 genes were downregulated, including more than 1300 abolished at the transcriptomic level. Hundreds of those downregulated genes were involved in the regulation of transcription, translation, and post-translational modifications and the organization and function of the nuclear chromosome, chromatin, and protein-DNA complex. DNA-binding elements in promoter regions of 130 dysregulated genes were predicted to be targeted by Cla4A domains. Samples of purified Cla4A extract were proven to bind promoter DNAs of 12 predicted genes involved in multiple stress-responsive pathways. Therefore, Cla4A acts as a novel regulator of genomic expression and stability and mediates gene expression networks required for insect-pathogenic fungal adaptations to the host and environment.

The impact of incorporating dried cafeteria leftover food on growing APRI rabbits productivity, profitability and meat quality.

This study was conducted to investigate the effect of dried cafeteria leftover food (DCLF) incorporation in growing APRI rabbit diets on nutrient digestibility, live body weight, body weight gain, carcass traits, meat quality, blood parameters, and economic efficiency. Thirty-six APRI rabbits (6 weeks of age), after weaning, were randomly divided into three groups (12 rabbits/group, 613 ± 5 g). During the experiment, rabbits aged 6-14 weeks participated in the experiment. An approximate isocaloric and isonitrogenous composition was achieved in three pelleted experimental diets. For the rabbit groups D1 (as the control), D2, and D3, DCLF was added at doses of 0%, 17.5%, and 35%, respectively. Digestibility trails were carried out at the end of the feeding experiment to evaluate the nutritional and feeding values of the experimental diets. To examine the characteristics of the carcasses, the quality of the meat, the histopathological analysis, and the blood plasma parameters, three rabbits were slaughtered from each group. The results showed that the feeding values (TDN and DE), which had the greatest values, as well as the digesting coefficients for dry matter, organic matter, crude fiber, and ether extract between the D1 and D3 groups were significantly different (p < 0.05). On the contrary, the curde protein, nitrogen-free extract, and digestible crude protein of the examined diets showed no signifcant differences compared with the control diet. The level of DCLF incorporation in diets significantly (p < 0.05) increased the live body weight at 14 weeks; with D3 recording the highest values (2130.9 g) when compared to the D1 and D2 groups. A similar pattern was observed for both daily increase and total body gain (6-14 weeks). Feed conversion recorded insignificant differences among the experimental diets respecting the period 6-14 weeks of age. DCLF levels had a significant effect on dressing and giblet percentages. The chemical composition and color parameters of rabbit meat were similar (p > 0.05), except for fat%, which had the highest value in the D3 group (p < 0.05) compared to the D1 and D2 groups. Economic efficiency was improved by using DLCF up to 35% in terms of gross margin and feed cost/kg gain. In conclusion, using DCLF in the feeding system of rabbits could enhance productive performance, carcass traits, meat quality, and physiological function, as well as improve economic efficiency.

The Association Between Bone Remodeling Markers, genes polymorphisms with Incidence of Osteopenia Among Young Saudi Female.

Osteopenia and osteoporosis, are prevalent skeletal systemic conditions, cause weaker bones and an increased risk of fragility fractures. This work is aimed to evaluate the relation between bone-remolding markers and genotypes of four single nucleotide polymorphisms in young Saudi females (rs2297480 of farnesyl diphosphate synthase (FDPS), rs3736228 of Low-density lipoprotein receptor-related protein 5 (LRP5), rs1234612 of sclerostin (SOST), and rs9934438 of Vitamin K epoxide reductase complex subunit 1  (VKORC1) ). For this purpose, 750 premenopausal females aged 18 to 40 years old, either university students, postgraduates, or university employees were recruited and divided into three groups according to bone mineral density BMD (g/cm2) divided by T score into osteoporosis (n = 12), osteopenia (n = 147), and normal (n = 591). Serum SOST, BALP, calcium, phosphate, ALP, albumin, beta-CTXs and human VDR levels were determined. TaqMan SNP Genotyping assays were used to genotype four polymorphisms using real-time PCR (applied biosystem). Results showed that BALP, CTX-1 and SOST were significantly higher in the osteoporosis and osteopenia groups than in the normal group. Bone mineral density readings were considerably lower in females with the GG genotype in FDPS rs2297480 and TT genotype in LRP5 rs3736228, which increase the risk for osteopenia by 3. 6-fold and 3. 06-fold than control respectively. Also, females with the TT genotype in LRP5 rs3736228 have decreased average values for Bone Mineral Density. In conclusion, the GG genotype of FDPS rs2297480 and the TT genotype of LRP5 rs3736228 was shown to be strongly associated with osteopenia in young Saudi females with low bone mineral density and SOST levels.

Does bevacizumab carry a hope for metastatic triple-negative breast cancer in the era of immunotherapy?

Triple-negative breast cancer (TNBC) has a very high rate of recurrence. Our aim is to investigate the efficacy of bevacizumab, platinum and paclitaxel as first-line in metastatic TNBC (mTNBC). This study included 54 female patients with mTNBC. They received bevacizumab, carboplatin and paclitaxel every 21 day for six cycles then who progressed shifted to second-line chemotherapy and the responders continue another two cycles. The median progression-free survival (PFS) was 27 months [95% confidence interval (CI), 17.019-36.981]. There were two factors that affect PFS; visceral only metastasis (hazard ratio, 0.23; P = 0.05) and performance status 0 (hazard ratio = 0.16; P = 0.02) with C-index 0.77. The median overall survival (OS) was 55 months (95% CI, 38.973-71.027). There were two factors that affect OS; type of presentation (hazard ratio = 7.91; P = 0.02) and performance status 0 (hazard ratio = 0.12; P = 0.01) with C-index 0.73. In the final evaluation, three factors have their print on achieving either stable disease (SD) or complete response (CR). Patients having ≤3 sites of metastasis odds ratio (OR) 3.92 (P = 0.02), patients with visceral only metastasis OR was 13.20 (P = 0.001), those with performance status 0 had the highest OR 19.5 (P = 0.001) and the percentage of this prediction was 64.8, 70.4 and 70.4%, respectively. Bevacizumab, carboplatin and paclitaxel were well tolerated, continuation of bevacizumab is recommended as long as SD or CR responses are obtained and tolerated.

Molecular Analysis of ZNF71 KRAB in Non-Small-Cell Lung Cancer.

Our previous study found that zinc finger protein 71 (ZNF71) mRNA expression was associated with chemosensitivity and its protein expression was prognostic of non-small-cell lung cancer (NSCLC). The Krüppel associated box (KRAB) transcriptional repression domain is commonly present in human zinc finger proteins, which are linked to imprinting, silencing of repetitive elements, proliferation, apoptosis, and cancer. This study revealed that ZNF71 KRAB had a significantly higher expression than the ZNF71 KRAB-less isoform in NSCLC tumors (n = 197) and cell lines (n = 117). Patients with higher ZNF71 KRAB expression had a significantly worse survival outcome than patients with lower ZNF71 KRAB expression (log-rank p = 0.04; hazard ratio (HR): 1.686 [1.026, 2.771]), whereas ZNF71 overall and KRAB-less expression levels were not prognostic in the same patient cohort. ZNF71 KRAB expression was associated with epithelial-to-mesenchymal transition (EMT) in both patient tumors and cell lines. ZNF71 KRAB was overexpressed in NSCLC cell lines resistant to docetaxel and paclitaxel treatment compared to chemo-sensitive cell lines, consistent with its association with poor prognosis in patients. Therefore, ZNF71 KRAB isoform is a more effective prognostic factor than ZNF71 overall and KRAB-less expression for NSCLC. Functional analysis using CRISPR-Cas9 and RNA interference (RNAi) screening data indicated that a knockdown/knockout of ZNF71 did not significantly affect NSCLC cell proliferation in vitro.

The effect of career guidance on undergraduate medical students' specialty preferences.

OBJECTIVE: To design and implement a career guidance programme for medical students, and to evaluate its effectiveness. METHODS: The quasi-experimental single-group study was conducted at the College of Medicine, Jouf University, Sakaka, Saudi Arabia, during the 2018-19 academic year. A modified Medical Career Development Inventory covering 5 areas was used to assess how much students' thinking or planning had improved after implementing the designed career guidance programme following Kern's 6 steps for course design. This was done through focussed group discussions. In the second phase, a comprehensive sample comprising 4th and 5th year male and female medical students was enrolled to assess their readiness, planning and choice of specialties, and to compare the same variables post-intervention. Data was analysed using SPSS 22. RESULTS: There were 50 students in the first phase; 30(60%) males and 20(40%) females. In phase II, there were 82 subjects; 47(57.3%) males and 35(42.7%) females. Pre-intervention mean result was 2.60±0.29 which increased post-intervention to 3.16±0.20 (p=0.018), suggesting an improved degree of vocational development. CONCLUSIONS: The career guidance programme significantly improved degree of vocational development and readiness to cope with developmental tasks encountered throughout a physician's career.

Ringworm in calves: risk factors, improved molecular diagnosis, and therapeutic efficacy of an Aloe vera gel extract.

BACKGROUND: Dermatophytosis in calves is a major public and veterinary health concern worldwide because of its zoonotic potential and associated economic losses in cattle farms. However, this condition has lacked adequate attention; thus, to develop effective control measures, we determined ringworm prevalence, risk factors, and the direct-sample nested PCR diagnostic indices compared with the conventional methods of dermatophytes identification. Moreover, the phenolic composition of an Aloe vera gel extract (AGE) and its in vitro and in vivo antidermatophytic activity were evaluated and compared with those of antifungal drugs. RESULTS: Of the 760 calves examined, 55.79% (424/760) showed ringworm lesions; 84.91% (360/424) were positive for fungal elements in direct-microscopy, and 79.72% (338/424) were positive in culture. Trichophyton verrucosum was the most frequently identified dermatophyte (90.24%). The risk of dermatophytosis was higher in 4-6-month-old vs. 1-month-old calves (60% vs. 41%), and in summer and winter compared with spring and autumn seasons (66 and 54% vs. 48%). Poor hygienic conditions, intensive breeding systems, animal raising for meat production, parasitic infestation, crossbreeding, and newly purchased animals were statistically significant risk factors for dermatophytosis. One-step PCR targeting the conserved regions of the 18S and 28S genes achieved unequivocal identification of T. verrucosum and T. mentagrophytes in hair samples. Nested-PCR exhibited an excellent performance in all tested diagnostic indices and increased the species-specific detection of dermatophytes by 20% compared with culture. Terbinafine and miconazole were the most active antifungal agents for dermatophytes. Gallic acid, caffeic acid, chlorogenic acid, cinnamic acid, aloe-Emodin, quercetin, and rutin were the major phenolic compounds of AGE, as assessed using high-performance liquid chromatography (HPLC). These compounds increased and synergized the antidermatophytic activity of AGE. The treated groups showed significantly lower clinical scores vs. the control group (P < 0.05). The calves were successfully treated with topical AGE (500 ppm), resulting in clinical and mycological cure within 14-28 days of the experiment; however, the recovery was achieved earlier in the topical miconazole 2% and AGE plus oral terbinafine groups. CONCLUSIONS: The nested PCR assay provided a rapid diagnostic tool for dermatophytosis and complemented the conventional methods for initiating targeted treatments for ringworm in calves. The recognized antidermatophytic potential of AGE is an advantageous addition to the therapeutic outcomes of commercial drugs.

Imidacloprid effects on acetylcholinesterase and nicotinic acetylcholine receptor in Apis mellifera. Experimental and molecular modeling approaches.

Although the neonicotinoid insecticides have good selectivity towards insects rather than vertebrates, they have severe effects on honeybee production and pollination activities. Therefore, the effects of imidacloprid (IMI), the most used neonicotinoid, on the two main bioreceptors, acetylcholinesterase (AChE) and nicotinic acetylcholine receptor alpha subunit (nAChRα1) of honeybees were examined to identify their roles in honeybee toxicity and possible binding sites which assist in selecting and designing neonicotinoids. In vivo, IMI showed a high inhibitory effect on AChE (IC50 5.63 mg/L); however, the effect was much lower in vitro experiment (IC50 719 mg/L). This result induced us to examine the IMI effect on AChE gene expression which revealed that the AChE-2 gene expression was severely affected by IMI explaining the observed high enzyme inhibition. In addition, although toxicity increased by increasing exposure to IMI (LC50 2.9 mg/L after 4h and 0.75 mg/L after 48h), AChE was not elevated (IC50 5.63 and 5.52 mg/L respectively). Besides, Despite resuming most enzyme activity (77% during 2 h and 84.14% after 4 h), a high mortality level was observed with LC50 2.9 mg/L. These results reinforced that the observed high toxicity is a multifactor process. Accordingly, Molecular modeling and docking of IMI into honeybee AChE and nAChRα1were also performed to examine their possible interactions and identify the important binding sites. Results models indicated that the first two binding sites in AChE were found in the esteratic subunit in the active site explaining the observed in vitro inhibition. In nAChRα1, four of the highest five free energy binding sites are located in the large TM3-TM4 loop and one in the extracellular loops. Consequently, the present work revealed that IMI toxicity is attributed to various factors including direct interaction with both AChE and nAChRα1 as well as downregulating AChE-2 gene expression.

Maximization of Biodiesel production from oil that produced during salmon smoking process with high amount of omega-3 by using homogenous catalyst.

The escalating global demand for oil, coupled with declining fossil fuel production, prompts the urgent exploration of renewable alternatives. To address this challenge, researchers are actively seeking environmentally friendly fuels like biodiesel. Among potential feedstocks, oil that is produced from salmon smoking process during industry emerges as a promising option. Smoked salmon oil could be a challenge when producing biodiesel due to its high content of omega-3 compounds. Using homogenous commercial catalyst from alkali to achieve the highest yield from salmon smoking oil was the aim of the current study. A Box-Behnken design and response surface methodology in Design Expert software (version 13) was used to study the effect of four main factors on the biodiesel yield from salmon smoking oil. The optimum biodiesel values were 70°C, 90 min, 0.753 wt.%, and 20 wt.% for temperature, reaction time, sodium hydroxide concentration, and methanol concentration, respectively. At these optimum values, the highest biodiesel production was 92.0% with fatty acid methyl ester contents of 83.4% and conversion efficiency of 77%. Thin-layer chromatography and thermal gravimetric analysis confirmed the successful production of biodiesel at optimized conditions. Using Aspen Plus simulation software confirmed the cost-effectiveness of the homogenous catalyst used for enhancing biodiesel production from salmon smoking oil.

Assessment of Patient Safety Culture Among Nurses Working at Tertiary Care Hospitals in Aljouf Region, Saudi Arabia.

INTRODUCTION: Patient safety is a fundamental element in healthcare quality and a major challenge in achieving universal health coverage, especially in low- and middle-income countries. The first step to improve patient safety is to evaluate the safety culture in hospitals. This study aimed to investigate the patient safety culture among nurses and determine the factors affecting it. METHODS: This cross-sectional study was conducted among 423 nurses working at tertiary care hospitals in the Al-Jouf region in Saudi Arabia. RESULTS: The highest score for patient safety among nurses was for teamwork within units (16.41 ± 2.44). The lowest score was for nonpunitive response to errors (5.87 ± 1.92). In addition, 83% of the participants did not report any events in the past 12 months. More perception of patient safety was significantly higher among females than males in dimensions of teamwork within units, frequency of events reported, and staffing. Furthermore, teamwork within units, management support for patient safety, staffing, non-punitive response to errors, and handoffs and transitions were significantly higher among participants in direct contact with patients. The Hospital Survey on Patient Safety Culture (HSOPSC) scale is significantly higher among non-Saudi nurses, nurses with bachelor's education, nurses with less working hours per week, and those who had training on patient safety. CONCLUSION: The current study showed that the majority of the participants did not report any events in the past 12 months. The highest score for patient safety culture dimensions among nurses was for teamwork within units while the lowest score was for nonpunitive response to errors.

Combination of Anatomical and Biological Factors to Predict Disease-Free Survival in Breast Cancer.

PURPOSE: The combination of anatomical and biological factors of breast cancer in a new staging system has a prognostic role. This study investigates the prognostic value of the Bioscore among patients with breast cancer with respect to disease-free survival (DFS). MATERIAL AND METHODS: This study included 317 patients with breast cancer who were identified between January 2015 and December 2018 at Clinical Oncology Department of Assiut University Hospital. Their cancer baseline characteristics were recorded: pathologic stage (PS), T stage (T), nodal stage (N), grade (G), estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor (HER2) status. Univariate and two multivariate analyses were performed to identify which of these variables are associated with DFS. Model performance was quantified using Harrell's concordance index (C-index), and the Akaike information criterion (AIC) was used to compare model fits. RESULTS: The significant factors in the univariate analysis were PS3, T2, T3, T4, N3, G2, G3, ER-negative, PR-negative, and HER2-negative. In the first multivariate analysis, PS3, G3, and ER-negative were the significant factors, and in the second multivariate analysis, T2, T4, N3, G3, and ER-negative were the significant factors. Two sets of models were built to determine the utility of combining variables. Models incorporating G and ER status had the highest C-index (0.72) for T + N + G + ER in comparison with (0.69) PS + G + ER and the lowest AIC (953.01) for T + N + G + ER and (966.9) for PS + G + ER. CONCLUSION: Using the Bioscore in breast cancer staging helps to identify patients at increased risk of recurrence. It provides more optimistic prognostic stratification than the anatomical staging alone for DFS.

Heart disease risk factors detection from electronic health records using advanced NLP and deep learning techniques.

Heart disease remains the major cause of death, despite recent improvements in prediction and prevention. Risk factor identification is the main step in diagnosing and preventing heart disease. Automatically detecting risk factors for heart disease in clinical notes can help with disease progression modeling and clinical decision-making. Many studies have attempted to detect risk factors for heart disease, but none have identified all risk factors. These studies have proposed hybrid systems that combine knowledge-driven and data-driven techniques, based on dictionaries, rules, and machine learning methods that require significant human effort. The National Center for Informatics for Integrating Biology and Beyond (i2b2) proposed a clinical natural language processing (NLP) challenge in 2014, with a track (track2) focused on detecting risk factors for heart disease risk factors in clinical notes over time. Clinical narratives provide a wealth of information that can be extracted using NLP and Deep Learning techniques. The objective of this paper is to improve on previous work in this area as part of the 2014 i2b2 challenge by identifying tags and attributes relevant to disease diagnosis, risk factors, and medications by providing advanced techniques of using stacked word embeddings. The i2b2 heart disease risk factors challenge dataset has shown significant improvement by using the approach of stacking embeddings, which combines various embeddings. Our model achieved an F1 score of 93.66% by using BERT and character embeddings (CHARACTER-BERT Embedding) stacking. The proposed model has significant results compared to all other models and systems that we developed for the 2014 i2b2 challenge.

Possible role of pomegranate fruit in reversing renal damage in rats exposed to Phenylhydrazine.

Background: Pomegranate granatum (molasses and peels) and its constituents showed protective effects against natural toxins such as phenylhydrazine (PHZ) as well as chemical toxicants such as arsenic, diazinon, and carbon tetrachloride. Aim: The current study aimed to assess the effect of pomegranate molasses (PM), white peel extract, and red peel extract on nephrotoxicity induced by PHZ. Methods: 80 male rats were divided into eight equal groups; a control group, PM pure group, white peel pomegranate pure group, red peel pomegranate pure group, PHZ group, PM + PHZ group, white peel pomegranate + PHZ group and red peel pomegranate + PHZ group. Kidney function, inflammation markers, antioxidant activities, and renal tissue histopathology were investigated. Results: The results revealed that PHZ group showed a significant increase in lactate Dehydrogenase (LDH), malondialdehyde (MDA), creatinine, uric acid, BUNBUN, C - reactive protein (CRP), tumor necrosis factor, thiobarbituric acid reactive substances (TBARSs), and total antioxidant capacity (TAC) with a significant decrease of catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD) as compared with a control group. Other pomegranate-treated and PHZ co-treated groups with pomegranate showed a significant decrease of LDH, MDA, creatinine, uric acid, BUN, tumor necrosis factor, TBARSs, and TAC with a significant increase of CAT, GPx, and SOD as compared with PHZ group. Conclusion: Collectively, our data suggest that red, white peels, and molasses have anti-toxic and anti-inflammatory effects on renal function and tissues.

Low baseline vitamin D levels increase the risk of bone metastases among females with breast cancer - Hospital based cohort study.

BACKGROUND: Serum vitamin D (Vit-D) has been linked to the development of breast cancer (BC); however, their effect on pathological features and outcomes is undetermined. The purpose of this study was to examine the prognostic significance of baseline Vit-D levels and their effect on clinical outcomes. METHODS: We evaluated baseline serum Vit-D levels and baseline clinic-pathological features of female patients with non-metastatic BC between October 2018 and December 2019. A low Vit-D level was described as less than 30 nanogram per liter (ng/l). Patients were observed for a median of 24 months. To evaluate relationships between qualitative variables, the chi-square test was used. The Kaplan-Meier technique was used for survival analysis, and the log-rank test was used to compare the two survival curves. Correlation analysis was also used to examine the link between Vit-D levels and clinical outcomes. RESULTS: The eligibility criteria were fulfilled by 221 patients. The median age of onset was (50.7). The median Vit-D level was (23.1 ng/l) with a range of (4-46 ng/l). Approximately half of the patients (56.5%) had Vit-D levels < 30 ng/l, with HER2 positive and triple negative (TNBC) patients having a greater proportion of low Vit-D levels (p = <0.001). Patients with low baseline Vit-D levels had a larger tumor size, more positive lymph nodes, and were diagnosed at a later stage. Following follow-up, Vit-D deficiency was associated with a significantly increased risk of bone metastases (HR 3.37, 95% CI 1.32-8.59, p = 0.006), and Vit-D levels were significantly correlated with disease-free survival (DFS) and overall survival (OS) (r = 0.850, r = 0.573, p < 0.00, p < 0.001, respectively). CONCLUSIONS: Low serum Vit-D is associated with advanced stage and adverse characteristics. It is more prevalent in HER-2 positive and TNBC patients; it increases the chance of bone metastases, and has a significant correlation with DFS and OS.

Assessment of Psychosocial Correlates and Associated Factors of Colorectal Cancer Screening among Southwestern Saudi Population: A Cross-Sectional Study.

Psychosocial correlates are one of the crucial determinants for the uptake of colorectal cancer (CRC) screening by the pre-eligible population. The present study aimed to identify the psychosocial correlates of colorectal cancer screening and determine their associated factors among the Saudi population in the Najran region, Saudi Arabia. Using a validated questionnaire, we assessed five constructs of psychosocial correlates of CRC screening among 790 participants aged 45 years and above. The five constructs were salience and coherence, cancer worries, perceived susceptibility, response efficacy, and social influence. Of the studied population, less than 50% agreed with most of the five constructs' statements, and 27.5% preferred to follow their family members' advice. Significantly higher mean scores (±SD) were identified for the male gender (7.38 ± 2.15, p = 0.027) and participants working in government sectors (7.60 ± 2.03, p = 0.027) in the cancer worries construct, while the mean (±SD) scores of perceived susceptibility were significantly higher among married participants (14.38 ± 4.10, p = 0.023) and smokers (14.95 ± 3.92, p = 0.041). Our survey results could help policymakers to implement focused health education programs for the pre-eligible population on the importance of the uptake of CRC screening. Furthermore, it is recommended to carry out exploratory mixed-method surveys in other regions of Saudi Arabia to understand the region's specific psychosocial correlates towards CRC screening.

Isolation and characterization of heavy-metal resistant microbes from roadside soil and phylloplane.

Contamination by heavy metals is one of the major environmental problems in many countries and these contaminants reach from various sources such as traffic cars and other activities. Soil and phylloplane samples were collected from eight traffic and two non-traffic sites in Sohag city, Egypt. Heavy metal contents of Cd²âº, Zn²âº and Pb²âº of soil and phylloplane samples were determined and revealed high levels of Zn²âº and Pb²âº in traffic samples. A total of 112 bacterial and 62 fungal isolates were obtained from soil and phylloplane. Bacterial isolates were characterized on the basis of morphological, physicochemical and biochemical characteristics; and 16S rRNA gene sequences. Fungal isolates were identified according to morphological characterization. Minimal inhibitory concentrations (MICs) of Cd²âº, Zn²âº and Pb²âº for each isolate were detected. All bacterial and fungal isolates demonstrated resistance to lead with MICs >0.528 mM and >0.211, respectively. Moreover, the maximum MICs of cadmium and zinc for bacteria were 0.821 mM and 1.471 mM, respectively, where as, MICs for fungi were 0.328 mM and 0.588 mM, respectively. The most resistant bacterial and fungal isolates were Pseudomonas aeruginosa RA65 and Penicillium corylophyllum, respectively. Therefore, P. aeruginosa RA65 was selected for further investigations. Growth curve study showed that 0.264 mM lead had no efficiently effect on the growth of P. aeruginosa RA65. Plasmid isolation evidenced by transformation studies indicated that P. aeruginosa RA65 harbored a single plasmid (~9.5 kb) which mediated heavy meal resistance. Consequently, these microbial isolates could be potentially used in bioremediation of heavy metal-contaminated environment.

Improvement of Zn (II) and Cd (II) Biosorption by Priestia megaterium PRJNA526404 Isolated from Agricultural Waste Water.

Heavy metals are considered as dangerous pollutants even in relatively low concentrations. Biosorption is an ecofriendly technology that uses microbial biomasses for adsorbing heavy metals from wastewater on their surfaces based on physicochemical pathways. Ten agricultural wastewater samples were collected from different sites in Sohag Governorate, Egypt. One hundred and nineteen zinc and cadmium-resistant bacterial isolates were recovered from the water samples. Interestingly, the isolate R1 was selected as the most resistant to Zn2+ and Cd2+. This isolate was morphologically and biochemically characterized and identified by sequencing of 16S rRNA gene as Priestia megaterium, and then deposited in the GenBank database under the accession number PRJNA526404. Studying the effects of pH and contact time on the biosorption process revealed that the maximum biosorption was achieved within 50 min at pH 7 and 8 for Zn2+ and Cd2+, respectively, by the living and lyophelized biomass of Priestia megaterium PRJNA526404. The preliminary characterization of the main chemical groups present on the cell wall, which are responsible for heavy metal biosorption, was performed by Infrared analysis (IR). Kinetics studies revealed that data were fitted towards the models hypothesized by Langmuir and Freundlich isotherm equations. The maximum capacity values (qmax) for biosorption of zinc and cadmium reached by using living and lyophelized biomass were 196.08; 227.27 and 178.57; 212.777 mg/g, respectively, and it was indicated that lyophilization improved efficiency of the biomass to heavy metals compared to living cells. The results indicated that Priestia megaterium PRJNA526404 had good application prospect in cadmium and zinc water remediation.

Characterization of BbKlf1 as a novel transcription factor vital for asexual and infection cycles of Beauveria bassiana.

The large family of C2H2-type zinc finger transcription factors (TFs) comprise the Kruppel-like factors (KLFs) that evolved relatively late in eukaryotes but remain unexplored in filamentous fungi. Here, we report that an orthologue (BbKlf1) of yeast Klf1 mediating cell wall integrity (CWI) is a wide-spectrum TF evidently localized in nucleus and cytoplasm in Beauveria bassiana. BbKlf1 features conserved domains and multiple DNA-binding motifs predicted to bind multiple promoter DNA fragments of target genes across asexual developmental and stress-responsive pathways. Despite limited impact on normal colony growth, deletion of Bbklf1 resulted in impaired CWI and hypersensitivity to Congo red-induced cell wall stress. Also, the deletion mutant was severely compromised in tolerance to oxidative and osmotic stresses, hyphal septation and differentiation, conidiation capacity (reduced by 95%), conidial quality (viability and hydrocarbon epitope pattern) and virulence. Importantly, these phenotypes correlated well with sharply repressed or nearly abolished expressions of those genes required for or involved in chitin biosynthesis, antioxidant activity, cell division and differentiation, aerial conidiation and conidial maturation. These findings indicate an essentiality of BbKlf1 for the asexual and insect-pathogenic lifecycles of B. bassiana and a novel scenario much beyond the yeast orthologue-mediated CWI, suggesting important roles of its orthologues in filamentous fungi.

Performance features and mortality prediction of the 4C Score early in COVID-19 infection: a retrospective study in Saudi Arabia.

The ISARIC4C consortium developed and internally validated the 4C Score for prediction of mortality only in hospitalized patients. We aimed to assess the validity of the 4C Score in mortality prediction of patients with COVID-19 who had been home isolated or hospitalized.This retrospective cross-sectional study was performed after the first wave of COVID-19. Data of all PCR-positive COVID-19 patients who had been discharged, hospitalized, or died were retrospectively analyzed. Patients were classified into four risk groups according to the 4C Mortality Score. A total of (506) patients were classified as follows: low (57.1%), intermediate (27.9%), high (13%), and very high (2%) risk groups. Clinical, radiological, and laboratory data were significantly more severe in the high and very high-risk groups compared with other groups (p<0.001 for all). Mortality rate was correctly estimated by the model with 71% sensitivity, 88.6% specificity, and area under the curve of 0.9. The mortality rate was underestimated among the very high-risk group (66.2% vs 90%). The odds of mortality were significantly greater in the presence of hypoxia (OR 2.6, 95% CI 1.5 to 4.6, p<0.001) and high respiratory rate (OR 5.3, 95% CI 1.6 to 17.9, p<0.007), C reactive protein (CRP) (OR 3.5, 95% CI 1.8 to 6.8, p<0.001), and blood urea nitrogen (BUN) (OR 1.9, 95% CI 1.3 to 3.1, p<0.002). Other components of the model had non-significant predictions. In conclusion, the 4C Mortality Score has good sensitivity and specificity in early risk stratification and mortality prediction of patient with COVID-19. Within the model, only hypoxia, tachypnea, high BUN, and CRP were the independent mortality predictors with the possibility of overlooking other important predictors.

Prevalence of multidrug resistant Salmonella spp. in dairy products with the evaluation of the inhibitory effects of ascorbic acid, pomegranate peel extract, and D-tryptophan against Salmonella growth in cheese.

The present study aimed at investigation of the prevalence and antimicrobial susceptibility of Salmonella spp. in the retailed dairies in Egypt. Besides, the inhibitory effects of some natural additives, including, ascorbic acid, pomegranate peel extract, and D-tryptophan against the isolated Salmonella were evaluated using soft cheese as a food matrix. To reach to this end, different Egyptian retail dairy products were investigated; 30 samples of each product were analyzed. Kariesh cheese samples had the highest Salmonella prevalence rate at 16.67%, followed by market raw milk, and bulk tank milk at 6.66% each, and white soft cheese at 3.33%. Serological examination exhibited 5 different Salmonella serotypes, namely S. Enteritidis, S. Typhimurium, S. Virchow, S. Larochelle, and S. Apeyeme. Antimicrobial susceptibility testing indicated that 100% of the isolates possessed resistance to erythromycin, oxacillin, and nalidixic acid. Some isolates of S. Typhimurium and S. Enteritidis were resistant to all 14 examined antibiotics. Isolates of S. Enteritidis obtained in this study were used to contaminate the freshly prepared soft cheese. Treatment of the artificially Salmonella-contaminated soft cheese with pomegranate peel extract, ascorbic acid, and D-tryptophan revealed a significant (P < 0.05) reduction in Salmonella growth in a dose-dependent manner. Therefore, the examined natural additives can be viewed as a promising new line of preservatives for dairy industry.