Identification of natural inhibitors targeting Trehalase of Anopheles funestus in the management of malaria: A Biocomputational assessment.

BACKGROUND OBJECTIVES: Anopheles funestus is playing an increasingly important role in malaria transmission in sub-Saharan Africa. Trehalase, an enzyme required for trehalose breakdown, is important for mosquito flight and stress adaptation. Hence, its inhibition has emerged as a promising malaria management strategy. METHODS: A collection of 1900 natural compounds from the ZINC database were screened against the 3D modeled structure of the A. funestus trehalase protein using in-silico tools. ADMET-AI, a web-based platform, was used to predict the absorption, distribution, metabolism, excretion, and toxicity (ADMET) properties of the selected compounds. RESULTS: Here in this study, we report 5 natural compounds namely, ZINC00488388, ZINC00488525, ZINC00488566, ZINC00488304, and ZINC00488456 demonstrated strong binding affinity to the trehalase protein. These compounds interacted with critical residues of the trehalase protein and exhibited good drug-like characteristics. INTERPRETATION CONCLUSION: These compounds show promise as trehalase protein inhibitors for malaria management. Nonetheless, additional experimental studies are required to optimize these compounds as potential trehalase inhibitors.

Staphylococcus aureus SigS Induces Expression of a Regulatory Protein Pair That Modulates Its mRNA Stability.

SigS is the sole extracytoplasmic function sigma factor in Staphylococcus aureus and is necessary for virulence, immune evasion, and adaptation to toxic chemicals and environmental stressors. Despite the contribution of SigS to a myriad of critical phenotypes, the downstream effectors of SigS-dependent pathogenesis, immune evasion, and stress adaptation remain elusive. To address this knowledge gap, we analyzed the S. aureus transcriptome following transient overexpression of SigS. We identified a bicistronic transcript, upregulated 1,000-fold, containing two midsized genes, each containing single domains of unknown function (DUFs). We renamed these genes SigS-regulated orfA (sroA) and SigS-regulated orfB (sroB). We demonstrated that SigS regulation of the sroAB operon is direct by using in vitro transcription analysis. Using Northern blot analysis, we also demonstrated that SroA and SroB have opposing autoregulatory functions on the transcriptional architecture of the sigS locus, with SroA stimulating SigS mRNA levels and SroB stimulating s750 (SigS antisense) levels. We hypothesized that these opposing regulatory effects were due to a direct interaction. We subsequently demonstrated a direct interaction between SroA and SroB using an in vivo surrogate genetics approach via bacterial adenylate cyclase-based two-hybrid (BACTH) analysis. We demonstrated that the SroA effect on SigS is at the posttranscriptional level of mRNA stability, highlighting a mechanism likely used by S. aureus to tightly control SigS levels. Finally, we demonstrate that the sroAB locus promotes virulence in a murine pneumonia model of infection. IMPORTANCE SigS is necessary for S. aureus virulence, immune evasion, and adaptation to chemical and environmental stressors. These processes are critically important for the ability of S. aureus to cause disease. However, the SigS-dependent transcriptome has not been identified, hindering our ability to identify downstream effectors of SigS that contribute to these pathogenic and adaptive phenotypes. Here, we identify a regulatory protein pair that is a major direct target of SigS, known as SroA and SroB. SroA also acts to stimulate SigS expression at the posttranscriptional level of RNA turnover, providing insight into intrinsically low levels of SigS. The discovery of SroA and SroB increases our understanding of SigS and the S. aureus pathogenesis process.

Novel Glu-based pyrazolo[3,4-d]pyrimidine analogues: design, synthesis and biological evaluation as DHFR and TS dual inhibitors.

A novel series of multifunctional pyrazolo[3,4-d]pyrimidine-based glutamate analogs (6a-l and 7a,b) have been designed and synthesized as antifolate anticancer agents. Among the tested compounds, 6i exhibited the most potent anti-proliferative activity towards NSCLC, CNS, Ovarian, Prostate, Colon, Melanoma, Breast, and Renal cancers with good to weak cytostatic activity and non-lethal actions. 6i demonstrated higher selectivity for cancer than normal cells. 6i could significantly increase the accumulation of S-phase cells during the cell cycle distribution of cancer cells with high potency in the induction of apoptosis. The results unveiled that 6i probably acts through dual inhibition of DHFR and TS enzymes (IC50 = 2.41 and 8.88 µM, correspondingly). Docking studies of 6i displayed that N1-p-bromophenyl and C3-Methyl groups participate in substantial hydrophobic interactions. The drug-likeness features inferred that 6i met the acceptance criteria of Pfizer. Taking together, 6i could be a promising prototype for further optimization as an effective anticancer drug.

Satisfaction and anxiety level during clinical training among nursing students.

BACKGROUND: Quality is a primary concern of health care agencies worldwide. A conducive clinical training environment is essential for nursing students to be capable of enhancing their learning experiences and achieving the desired training outcomes. AIM: This study aimed to examine the satisfaction and anxiety levels during clinical training among nursing students. TYPE OF STUDY: A descriptive -analytical cross-sectional study design was utilized. The research was conducted at the Faculty of Nursing, Assiut University and Colleges of Applied Medical Sciences in Alnamas and Bisha, University of Bisha. Sampling method: A convenience sampling technique was used. SAMPLE SIZE: a sample of 1052 undergraduate nursing students. The data was gathered via a structured questionnaire including the socio-demographic characteristics and nursing students' satisfaction with the hospital and laboratory training. Additionally, Self-Rating Anxiety Scale (SAS) was adopted to measure the anxiety level. RESULTS: The mean age of the studied sample was 21.9 ± 1.83 years, and 56.9% are females. Moreover, 90.1% & 76.4% of the nursing students were satisfied with their hospital and laboratory training. Furthermore, 61.1% & 54.8% of the students had mild levels of anxiety regarding their hospital training and laboratory training, respectively. CONCLUSION: The undergraduate nursing students had a high level of satisfaction with their clinical training at the hospitals and laboratories. Moreover, they had mild anxiety related to hospital and laboratory clinical training. RECOMMENDATIONS: Developing clinical orientation and training programs and improvement strategies to enhance the effectiveness of the clinical training environment. The establishment of a modern, tastefully designed, and fully stocked skill lab for the college's student training should receive more attention. CLINICAL RELEVANCE: Through the provision of ongoing education about different method of practice, nursing was intended to shape future professional nurses who master core competencies of the profession. Organizations may benefit from developing a comprehensive strategy to achieve an effective teaching program.

Characterization of Bioactive Compounds from Acacia concinna and Citrus limon, Silver Nanoparticles' Production by A. concinna Extract, and Their Biological Properties.

The applications of bioactive compounds from medicinal plants as therapeutic drugs are largely increasing. The present study selected the bioactive compounds from Acacia concinna (A. concinna) and Citrus limon (C. limon) to assess their phytochemicals, proteins, and biological activity. The plant material was collected, and extraction performed as per the standard procedure. Qualitative analysis was undertaken, and identification of functional organic groups was performed by FTIR and HPLC. Antibacterial, anticancer, antioxidant, antihyperglycemic, antihyperlipidemic, and inhibition kinetics studies for enzymes were performed to assess the different biological activities. Flavonoids and phenols were present in a significant amount in both the selected plants. A. concinna showed significant antimicrobial activity against Z. mobilis, E. coli, and S. aureus, with minimum inhibition zones (MIZ) of 24, 22, and 20 mm, respectively. C. limon strongly inhibited all the tested pathogenic bacteria with maximum and minimum MIZ of 32 and 17 mm. A. concinna silver nanoparticles also exhibited potent antimicrobial activity. Both extracts showed substantial antioxidant, antihyperlipidemic, antidiabetic, anticancer (MCF-7), and anti-urease (antiulcer) properties. To conclude, these plants can be used to treat hyperlipidemia, diabetes, cancer, and gastrointestinal ulcers. They can also serve as antimicrobial and antioxidant agents. Thus, the studied plants must be exploited cost-effectively to generate therapeutic drugs for various diseases.

Effectiveness of Pre-Transplant Screening for High-Priority Multidrug-Resistant Pathogens on Pre-Engraftment Infections After Hematopoietic Stem Cell Transplantation.

Objective: Owing to the rising incidence of multidrug-resistant organisms (MDRO) and the high mortality rates associated with such bacterial infections post-hematopoietic stem cell transplantation (HSCT), we investigated the MDRO colonization rate prior to transplantation using an active surveillance approach and determined its impact on subsequent infection during the pre-engraftment period. Methods: A single-center observational study was conducted, and surveillance cultures from multiple body sites, including the rectum, nasal cavity, and groin, were performed at admission to determine MDRO colonization. Serological tests were used to detect certain viruses and toxoplasmosis before HSCT. Results: In the pre-transplant setting, 59 MDRO were recovered from the 40 HSCT recipients. Of the 59 isolates recovered from one or more body sites, 29 were positive for methicillin-resistant Staphylococcus aureus (MRSA), 7 for carbapenem-resistant Enterobacterales (CRE), and 23 were positive for extended-spectrum ß-lactamase (ESBLs). Serological assessment before HSCT revealed active or reactivation of latent infection with cytomegalovirus (7.5%), Epstein-Barr virus (EBV; 5%), and Toxoplasma gondii (2.5%) among HSCT patients. In terms of factors associated with pre-engraftment infections, the type of transplant (p=0.04) was statistically significant, whereas other factors, such as age, sex, and underlying conditions, were not. In post-transplant settings, bloodstream infections (BSIs) were documented in 2 allogeneic HSCT patients (5%), and the isolated microorganisms were ESBL-producing E. coli and non-MDR Acinetobacter baumannii. Conclusion: Active screening cultures are a helpful tool for identifying patients colonized by MDRO or relevant viruses before HSCT, and for predicting those at risk of developing subsequent pre-engraftment infections. Additionally, active screening may aid in predicting those who are likely to develop subsequent pre-engraftment infections Our findings highlight the importance of pre-transplant screening for high-priority multidrug-resistant pathogens and the application of infection control interventions after HSCT.

Antibacterial and hemocompatibility potentials of nano-gold-cored alginate preparation against anaerobic bacteria from acne vulgaris.

Acne is a prevalent dermatological disease, with high global incidence, and is a health menace. The current study aimed to isolate and characterize the anaerobic bacteria responsible for the condition. Causes of a total of 70 acne-based bacterium isolates obtained from patients of mild, moderate, and severe acne, 24 were Clostridium innocuum, 21 were Lactobacillus plantarum, 13 were Anaerococcus prevotii, and 12 were Peptoniphilus asaccharolyticus. Nearly 69% of males were suffering, while the rest were females at 31%. The 15-30 years old age group was the most affected. The gold/alginate nanoparticles' nanopreparation (GANPs) produced from chloroauric acid and sodium alginate was an effective treatment against the acne conditions under the experimental conditions. The nanopreparation exhibited significant inhibitory activity against anaerobic bacterial isolates, with a minimum inhibitory concentration of 200 µg/ml for A. prevotii and P. asaccharolyticus, and 400 µg/ml for C. innocuum and L. plantarum. The in vitro efficacy of the GANPs on human blood parameters was also assessed. The concurrent results suggested potential antibacterial activity and hemocompatibility of the product, which has promise to be used as a successful antibacterial agent for acne.

Bacterial cellulase production via co-fermentation of paddy straw and Litchi waste and its stability assessment in the presence of ZnMg mixed-phase hydroxide-based nanocomposite derived from Litchi chinensis seeds.

Co-fermentation via co-cultured bacterial microorganisms to develop enzymes in solid-state fermentation (SSF) is a promising approach. This strategy is imperative in a series of sustainable and effective approaches due to superior microbial growth and the use of a combination of inexpensive feedstocks for enzyme production wherein mutually participating enzyme-producing microbial communities are employed. Moreover, the addition of nanomaterials to this technique may aid in its prominent advantage of enhancing enzyme production. This strategy may be able to decrease the overall cost of the bioprocessing to produce enzymes by further implementing biogenic, route-derived nanomaterials as catalysts. Therefore, the present study attempts to explore endoglucanase (EG) production using a bacterial coculture system by employing two different bacterial strains, namely, Bacillus subtilis and Serratia marcescens under SSF in the presence of a ZnMg hydroxide-based nanocomposite as a nanocatalyst. The nanocatalyst based on ZnMg hydroxide has been prepared via green synthesis using Litchi waste seed, while SSF for EG production has been conducted using cofermentation of litchi seed (Ls) and paddy straw (Ps) waste. Under an optimized substrate concentration ratio of 5:6 Ps:Ls and in the presence of 2.0 mg of nanocatalyst, the cocultured bacterial system produced 1.6 IU/mL of EG enzyme, which was ~1.33 fold higher as compared to the control. Additionally, the same enzyme showed its stability for 135 min in the presence of 1.0 mg of nanocatalyst at 38 °C. The nanocatalyst has been synthesized using the green method, wherein waste litchi seed is used as a reducing agent, and the nanocatalyst could be employed to improve the production and functional stability of crude enzymes. The findings of the present study may have significant application in lignocellulosic-based biorefinaries and cellulosic waste management.

Rice straw derived graphene-silica based nanocomposite and its application in improved co-fermentative microbial enzyme production and functional stability.

Cellulases are the one of the most highly demanded industrial biocatalysts due to their versatile applications, such as in the biorefinery industry. However, relatively poor efficiency and high production costs are included as the key industrial constraints that hinder enzyme production and utilization at economic scale. Furthermore, the production and functional efficiency of the ß-glucosidase (BGL) enzyme is usually found to be relatively low among the cellulase cocktail produced. Thus, the current study focuses on fungi-mediated improvement of BGL enzyme in the presence of a rice straw-derived graphene-silica-based nanocomposite (GSNCs), which has been characterized using various techniques to analyze its physicochemical properties. Under optimized conditions of solid-state fermentation (SSF), co-fermentation using co-cultured cellulolytic enzyme has been done, and maximum enzyme production of 42 IU/gds FP, 142 IU/gds BGL, and 103 IU/gds EG have been achieved at a 5 mg concentration of GSNCs. Moreover, at a 2.5 mg concentration of nanocatalyst, the BGL enzyme showed its thermal stability at 60°C and 70 °C by holding its half-life relative activity for 7 h, while the same enzyme demonstrated pH stability at pH 8.0 and 9.0 for the 10 h. This thermoalkali BGL enzyme might be useful for the long-term bioconversion of cellulosic biomass into sugar.

Polyphenol Fingerprint, Biological Activities, and In Silico Studies of the Medicinal Plant Cistus parviflorus L. Extract.

Cistus parviflorus L. (Cistaceae) is a medicinal plant with several folkloric applications, including being used for urinary tract infections and as a food additive. In this study, the polyphenolic diversity and the antioxidant, antidiabetic, and antimicrobial activities of the C. parviflorus methanolic extract were evaluated. Spectrophotometric and HPLC-based analyses using standard polyphenolic compounds were conducted to measure the phenolics and flavonoids in the plant extract. The in vitro DPPH, ORAC, FRAP, and α-glucosidase assays were used to evaluate the plant's antioxidant and antidiabetic activities. Furthermore, disc diffusion and MIC-based microdilution tests were applied to evaluate the antimicrobial activity of the plant against broad-spectrum microorganisms. The analysis revealed the existence of high phenolic and flavonoid quantities that were measured at 302.59 ± 0.6 µg GAE and 134.3 ± 0.5 µg RE, respectively. The HPLC-based analysis revealed the existence of 18 phenolic acids and 8 flavonoids. The major phenolic acid was ellagic acid (169.03 ppm), while catechin was the major flavonoid (91.80 ppm). Remarkable antioxidant activity was measured using three different assays: DPPH, ORAC, and FRAP. Furthermore, strong inhibition of α-glucosidase compared to acarbose was recorded for the plant extract (IC50 0.924 ± 0.6). The results showed that C. parviflorus's extract had a strong anti-Escherichia coli effect with MIC value of 0.98 µg\mL and IZD value of 32.2 ± 0.58 mm compared to 25.3 ± 0.18 mm for gentamycin, the positive control. Moreover, Aspergillus niger, Aspergillus fumigatus, Staphylococcus aureus, Streptococcus pyogenes, and Salmonella typhimurium all showed significant growth inhibition in response to the extract, a result that may be related to the use of the plant in traditional medicine to treat urinary tract infections. The docking study indicated the higher binding affinity of the major identified compounds, i.e., ellagic acid, rutin, naringin, catechin, and punicalagin, to the S. aureus gyrase-DNA complex, which might suggest the possible mechanisms of the plant as antimicrobial agents.

Evaluation of the Antibacterial Properties of Iron Oxide, Polyethylene Glycol, and Gentamicin Conjugated Nanoparticles against Some Multidrug-Resistant Bacteria.

Antibacterial resistance is observed as a public health issue around the world. Every day, new resistance mechanisms appear and spread over the world. For that reason, it is imperative to improve the treatment schemes that have been developed to treat infections caused by wound infections, for instance, Staphylococcus epidermidis (S. epidermidis), Proteus mirabilis (P. mirabilis), and Acinetobacter baumannii (A. baumannii). In this case, we proposed a method that involves mixing the Gentamicin (Gen) with iron oxide nanoparticles (Fe3O4 NPs) and a polymer (polyethylene glycol (PEG)) with Fe3O4 NPs. X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), energy dispersive X-ray (EDX), scanning electron microscope (SEM), and transmission electron microscope (TEM) were used to characterize Fe3O4 NPs. Zeta potential and dynamic light scattering (DLS) were also assessed. The antibacterial activity of Fe3O4 NPs, Fe3O4 NPs+PEG, Fe3O4 NPs+Gen, and Fe3O4 NPs+PEG+Gen composites was investigated. The results showed a significant improvement in the antibacterial activity of nanoparticles against bacterial isolates, especially for the Fe3O4 NPs+PEG+Gen as the diameter of the inhibition zone reached 26.33 ± 0.57 mm for A. baumannii, 25.66 ± 0.57 mm for P. mirabilis, and 23.66 ± 0.57 mm for S. epidermidis. The Fe3O4 NPs, Fe3O4 NPs+PEG, Fe3O4+Gen, and Fe3O4+PEG+Gen also showed effectiveness against the biofilm produced by these isolated bacteria. The minimum inhibitory concentration (MIC) of Fe3O4 NPs for S. epidermidis was 25 µg mL-1 and for P. mirabilis and A. baumannii was 50 µg mL-1. The findings suggest that the prepared nanoparticles could be potential therapeutic options for treating wound infections caused by S. epidermidis, P. mirabilis, and A. baumannii.

Green Synthesis and Characterization of Iron Nanoparticles Synthesized from Aqueous Leaf Extract of Vitex leucoxylon and Its Biomedical Applications.

The cold extraction method was used to obtain the aqueous extract of Vitex leucoxylon leaves in a ratio of 1:10. Iron nanoparticles (FeNPs) were synthesized using aqueous leaf extract of V. leucoxylon as a reducing agent. The phytoreducing approach was used to make FeNPs by mixing 1 mL of plant extract with 1 mM of ferric sulfate. Scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), Ultraviolet-visible spectroscopy (UV-Vis), and energy-dispersive X-ray spectroscopy were used to examine the synthesized FeNPs. The reducing reaction was shown by a change in the color of the solution, and the formation of black color confirms that FeNPs have been formed. The greatest absorption peak (max) was found at 395 nm in UV-Vis spectral analysis. The FTIR spectra of V. leucoxylon aqueous leaf extract showed shifts in some peaks, namely 923.96 cm-1 and 1709.89 cm-1, with functional groups carboxylic acids, unsaturated aldehydes, and ketones, which were lacking in the FTIR spectra of FeNPs and are responsible for FeNPs formation. FeNPs with diameters between 45 and 100 nm were observed in SEM images. The creation of FeNPs was confirmed by EDX, which shows a strong signal in the metallic iron region at 6-8 Kev. XRD revealed a crystalline nature and an average diameter of 136.43 nm. Antioxidant, anti-inflammatory, cytotoxic, and wound healing in vitro tests reported significant activity of the FeNPs. The cumulative findings of the present study indicate that the green synthesis of FeNPs boosts its biological activity and may serve as a possible dermal wound-healing agent and cytotoxic agent against cancer. Future study is needed on the identification of mechanisms involved in the synthesis of FeNPs by V. leucoxylon and its biomedical applications.

Biodegradation of Azo Dye Methyl Red by Pseudomonas aeruginosa: Optimization of Process Conditions.

Water pollution due to textile dyes is a serious threat to every life form. Bacteria can degrade and detoxify toxic dyes present in textile effluents and wastewater. The present study aimed to evaluate the degradation potential of eleven bacterial strains for azo dye methyl red. The optimum degradation efficiency was obtained using P. aeruginosa. It was found from initial screening results that P. aeruginosa is the most potent strain with 81.49% degradation activity and hence it was subsequently used in other degradation experiments. To optimize the degradation conditions, a number of experiments were conducted where only one variable was varied at a time and where maximum degradation was observed at 20 ppm dye concentration, 1666.67 mg/L glucose concentration, 666.66 mg/L sodium chloride concentration, pH 9, temperature 40 °C, 1000 mg/L urea concentration, 3 days incubation period, and 66.66 mg/L hydroquinone (redox mediator). The interactive effect of pH, incubation time, temperature, and dye concentration in a second-order quadratic optimization of process conditions was found to further enhance the biodegradation efficiency of P. aeruginosa by 88.37%. The metabolites of the aliquot mixture of the optimized conditions were analyzed using Fourier transform infrared (FTIR), GC-MS, proton, and carbon 13 Nuclear Magnetic Resonance (NMR) spectroscopic techniques. FTIR results confirmed the reduction of the azo bond of methyl red. The Gas Chromatography-Mass Spectrometry (GC-MS) results revealed that the degraded dye contains benzoic acid and o-xylene as the predominant constituents. Even benzoic acid was isolated from the silica gel column and identified by 1H and 13C NMR spectroscopy. These results indicated that P. aeruginosa can be utilized as an efficient strain for the detoxification and remediation of industrial wastewater containing methyl red and other azo dyes.

Clinical Prevalence, Antibiogram Profiling and Gompertz Growth Kinetics of Resistant Staphylococcus epidermidis Treated with Nanoparticles of Rosin Extracted from Pinus roxburghii.

The rise of methicillin-resistant Staphylococcus epidermidis (MRSE) makes it difficult to treat infections that increase morbidity and mortality rates in various parts of the world. The study's objectives include identifying the clinical prevalence, antibiogram profile, and Gompertz growth kinetics of MRSE treated with synthetically created nanoparticles of rosin obtained from Pinus roxburghii. A total of 64 of 200 clinical isolates of S. epidermidis (32% of the total) displayed sensitivity (40.62%) and resistance (59.37%) to seven different antibiotic classes. The most sensitive patterns of antibiotic resistance were seen in 20 (78.95%) and 24 (94.74%) isolates of MRSE against piperacillin/tazobactam and cephradine, respectively. Fosfomycine was found to be the most effective antibiotic against MRSE in 34 (89.47%) isolates, followed by amoxicillin. Successfully produced, described, and used against MRSE were rosin maleic anhydride nanoparticles with a size range of 250 nm to 350 nm. Five different concentrations of 25, 50, 75, 100, and 150 mg mL-1 rosin maleic anhydride nanoparticles were investigated to treat MRSE resistance. According to Gompertz growth kinetics, the maximal growth response was 32.54% higher and the lag phase was also 10.26% longer compared to the control when the amount of rosin maleic anhydride nanoparticles was increased in the MRSE. Following the application of rosin maleic anhydride nanoparticles, the growth period is extended from 6 to 8 h. A potential mechanism for cell disintegration and distortion is put forth. This investigation came to the conclusion that rosin maleic anhydride nanoparticles better interfere with the surface of MRSE and demonstrated a preferred bacteriostatic action.

Green Synthesis of Silver Nanoparticles Using Aqueous Citrus limon Zest Extract: Characterization and Evaluation of Their Antioxidant and Antimicrobial Properties.

The current work concentrated on the green synthesis of silver nanoparticles (AgNPs) through the use of aqueous Citruslimon zest extract, optimizing the different experimental factors required for the formation and stability of AgNPs. The preparation of nanoparticles was confirmed by the observation of the color change of the mixture of silver nitrate, after the addition of the plant extract, from yellow to a reddish-brown colloidal suspension and was established by detecting the surface plasmon resonance band at 535.5 nm, utilizing UV-Visible analysis. The optimum conditions were found to be 1 mM of silver nitrate concentration, a 1:9 ratio extract of the mixture, and a 4 h incubation period. Fourier transform infrared spectroscopy spectrum indicated that the phytochemicals compounds present in Citrus limon zest extract had a fundamental effect on the production of AgNPs as a bio-reducing agent. The morphology, size, and elemental composition of AgNPs were investigated by zeta potential (ZP), dynamic light scattering (DLS), SEM, EDX, X-ray diffraction (XRD), and transmission electron microscopy (TEM) analysis, which showed crystalline spherical silver nanoparticles. In addition, the antimicrobial and antioxidant properties of this bioactive silver nanoparticle were also investigated. The AgNPs showed excellent antibacterial activity against one Gram-negative pathogens bacteria, Escherichia coli, and one Gram-positive bacteria, Staphylococcus aureus, as well as antifungal activity against Candida albicans. The obtained results indicate that the antioxidant activity of this nanoparticle is significant. This bioactive silver nanoparticle can be used in biomedical and pharmacological fields.

Computational high-throughput screening and in vitro approaches identify CB-006-3; A novel PI3K-BRAFV600E dual targeted inhibitor against melanoma.

Malignant melanoma is characterized by both genetic and molecular alterations that activate phosphoinositide 3-kinase (PI3K), and RAS/BRAF pathways. In this work, through diversity-based high-throughput virtual screening we identified a lead molecule that selectively targets PI3K and BRAFV600E kinases. Computational screening, Molecular dynamics simulation and MMPBSA calculations were performed. PI3K and BRAFV600E kinase inhibition was done. A375 and G-361 cells were used for in vitro cellular analysis to determine antiproliferative effects, annexin V binding, nuclear fragmentation and cell cycle analysis. Computational screening of small molecules indicates compound CB-006-3 selectively targets PI3KCG (gamma subunit), PI3KCD (delta subunit) and BRAFV600E. Molecular dynamics simulation and MMPBSA bases binding free energy calculations predict a stable binding of CB-006-3 to the active sites of PI3K and BRAFV600E. The compound effectively inhibited PI3KCG, PI3KCD and BRAFV600E kinases with respective IC50 values of 75.80, 160.10 and 70.84 nM. CB-006-3 controlled the proliferation of A375 and G-361 cells with GI50 values of 223.3 and 143.6 nM, respectively. A dose dependent increase in apoptotic cell population and sub G0/G1 phase of cell cycle were also observed with the compound treatment in addition to observed nuclear fragmentation in these cells. Furthermore, CB-006-3 inhibited BRAFV600E, PI3KCD and PI3KCG in both melanoma cells. Collectively, based on the computational modeling and in vitro validations, we propose CB-006-3 as a lead candidate for selectively targeting PI3K and mutant BRAFV600E to inhibit melanoma cell proliferation. Further experimental validations, including pharmacokinetic evaluations in mouse models will identify the druggability of the proposed lead candidate for further development as a therapeutic agent for treating melanoma.

Autoimmune thyroiditis associated with Sanjad-Sakati syndrome: A call for regular thyroid screening.

Sanjad-Sakati Syndrome (SSS) is a rare autosomal recessive disorder characterized by congenital hypoparathyroidism, growth retardation and dysmorphism. Thyroid status of patients with SSS has not been widely explored. Therefore, we aimed to review the occurrence of autoimmune thyroiditis, which is commonly associated with other genetic disorders, in SSS. A retrospective hospital based study was conducted at King Khalid University Hospital, Riyadh, Saudi Arabia, to determine the thyroid status of patients with SSS attending the hospital between 1990 and 2015. Data were extracted from the medical records of patients diagnosed with Sanjad-Sakati syndrome with special emphasis on the clinical features, thyroid function, thyroid antibodies, molecular studies and other relevant investigations. A total of 18 patients with a diagnosis of Sanjad-Sakati Syndrome based on typical clinical features and low parathyroid hormone, were evaluated. Furthermore, molecular study was available on 15 patients; all had homozygous deletion of 12 bp (155-166) in exon 3 of the TCBE gene. In 6 patients the thyroid functions were abnormal (one patient with overt hypothyroidism and five patients with sub clinical hypothyroidism). Thyroid autoantibodies were positive in 4 patients. In conclusion, one third of this cohort with SSS had abnormal thyroid function test attributed mainly to autoimmune thyroiditis. Therefore, we recommend routine screening of patients with SSS for thyroid function and autoimmune antibodies during follow up.