In silico approaches for the identification of potential allergens among hypothetical proteins from Alternaria alternata and its functional annotation.

Direct exposure to the fungal species Alternaria alternata is a major risk factor for the development of asthma, allergic rhinitis, and inflammation. As of November 23rd 2020, the NCBI protein database showed 11,227 proteins from A. alternata genome as hypothetical proteins (HPs). Allergens are the main causative of several life-threatening diseases, especially in fungal infections. Therefore, the main aim of the study is to identify the potentially allergenic inducible proteins from the HPs in A. alternata and their associated functional assignment for the complete understanding of the complex biological systems at the molecular level. AlgPred and Structural Database of Allergenic Proteins (SDAP) were used for the prediction of potential allergens from the HPs of A. alternata. While analyzing the proteome data, 29 potential allergens were predicted by AlgPred and further screening in SDAP confirmed the allergic response of 10 proteins. Extensive bioinformatics tools including protein family classification, sequence-function relationship, protein motif discovery, pathway interactions, and intrinsic features from the amino acid sequence were used to successfully predict the probable functions of the 10 HPs. The functions of the HPs are characterized as chitin-binding, ribosomal protein P1, thaumatin, glycosyl hydrolase, and NOB1 proteins. The subcellular localization and signal peptide prediction of these 10 proteins has further provided additional information on localization and function. The allergens prediction and functional annotation of the 10 proteins may facilitate a better understanding of the allergenic mechanism of A. alternata in asthma and other diseases. The functional domain level insights and predicted structural features of the allergenic proteins help to understand the pathogenesis and host immune tolerance. The outcomes of the study would aid in the development of specific drugs to combat A. alternata infections.

Prevalence of Smear-Positive, Rifampicin-Resistant Mycobacterium tuberculosis and Related Factors Among Residents with Cough in Northern Ethiopian Refugee Health Facilities.

Purpose: To ascertain the prevalence of Mycobacterium tuberculosis (M.tb) among refugees suspected of tuberculosis (TB) and related risk factors, including smear-positive and Rifampicin-resistant M.tb. Methods: A cross-sectional study was conducted between January 2020 and May 2020 among 384 refugees in four refugee camps in Northwest Tigray, Ethiopia. Socio-demographic and clinical data were collected from refugees with a history of cough for more than two weeks prospectively. Spot-spot sputum samples were collected and transported in an ice box to the Shire Suhul Hospital Microbiology laboratory; and then examined using a Fluorescent Microscope. All smear-positive samples were further processed by GeneXpert to detect Rifampicin-resistant MTB. Data were analyzed using SPSS version 21 and a p-value <0.05 was considered statistically significant. Results: The overall prevalence of smear-positive PTB infection was 5.5% (21/384), but No TB case was resistant to Rifampicin detected by GeneXpert MTB/RIF assay. About 70% of the smear-positive pulmonary TB identified were females. Five (23.8%) of the smear-positive pulmonary tuberculosis cases were co-infected by HIV. Sharing of drink and food materials (AOR = 4.36, 95% CI = 1.19-15.89), active TB contact (AOR 7.24, 95% CI = 1.62-32.125), BMI (AOR = 5.23, 95% CI = 1.28-21.29), opening window practice (AOR = 4.32, 95% CI = 1.02-18.30) and HIV status (AOR = 9.36, 95% CI = 1.64-53.35) were statistically significant predisposing factors. Conclusion: The prevalence of smear-positive pulmonary TB among northwest Tigray refugee camps was still high. The prevalence of TB/HIV co-infection was also high. Minimizing close contact with active TB cases, reducing malnutrition, rapid TB/HIV screening, and establishing a ventilation system can reduce the transmission of TB among refugees.

Divergent Responses of Hydrophilic CdSe and CdSe@CdS Core-Shell Nanocrystals in Apoptosis and In Vitro Cancer Cell Imaging: A Comparative Analysis.

With their distinctive core-shell design, core-shell nanocrystals have drawn interest in catalysis, medicinal research, and nanotechnology. These nanocrystals have a variety of characteristics and possible uses. The application of core-shell nanocrystals offers significant potential in increasing diagnostic and therapeutic approaches for cancer research in apoptosis and in vitro cancer cell imaging. In the present study, we investigated the fluorescence behavior of hydrophilic CdSe (core-only) and CdSe@CdS (core-shell) nanocrystals (NCs) and their potential in cancer cell imaging. The addition of a CdS coating to CdSe NCs increased the fluorescence intensity tenfold. The successful fabrication of core-shell CdSe@CdS nanocrystals was proven by a larger particle size (evaluated via DLS and TEM) and their XRD pattern and surface morphology compared to CdSe (core-only) NCs. When these NCs were used for bioimaging in MCF-7 and HEK-293 cell lines, they demonstrated excellent cellular uptake due to higher fluorescence intensity within cancerous cells than normal cells. Comparative cytotoxicity studies revealed that CdSe NCs were more toxic to all three cell lines (HEK-293, MCF-7, and HeLa) than CdSe@CdS core-shell structures. Furthermore, a decrease in mitochondrial membrane potential and intracellular ROS production supported NCs inducing oxidative stress, which led to apoptosis via the mitochondria-mediated pathway. Increased cytochrome c levels, regulation of pro-apoptotic gene expression (e.g., p53, Bax), and down-regulation of Bcl-2 all suggested cellular apoptosis occurred via the intrinsic pathway. Significantly, at an equivalent dose of core-shell NCs, core-only NCs induced more oxidative stress, resulting in increased apoptosis. These findings shed light on the role of a CdS surface coating in reducing free radical release, decreasing cytotoxicity, and improving fluorescence, advancing the field of cell imaging.

Single-Step Synthesis of Ag Hexagonal Nanoplate-Decorated Reduced Graphene Oxide and Its Cytotoxicity Studies.

Graphene-based Ag nanocomposites are of specific interest because of their unique properties and applications, especially in the field of cytotoxicity. However, developing a simple method to synthesize reduced graphene oxide (rGO)/silver hexagonal nanoplate (Ag HNPT) (rGO-Ag HNPT) nanocomposites with well-defined morphology has been believed to be a major challenge. In this work, a facile, robust, and single-step synthesis method was developed to prepare silver-graphene (rGO-Ag HNPT) nanocomposites with hexagonal-structured silver nanoplates without any templates. The primary characterizations of the synthesized nanocomposite were done using a UV-visible spectrophotometer, X-ray diffraction (XRD), and Raman spectroscopy. The formation of uniformed hexagonal-shaped Ag nanoplates was confirmed by high-resolution transmission electron microscopy (HR-TEM), and the elemental composition was confirmed using energy dispersive X-ray analysis (EDX). With SiHa cervical cancer cells, the short-term in vitro cytotoxicity of the as-synthesized rGO-Ag HNPTs was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The anticancer response of the rGO-Ag HNPTs was investigated using an MTT assay.

Role and mechanistic actions of protein kinase inhibitors as an effective drug target for cancer and COVID.

Kinases can be grouped into 20 families which play a vital role as a regulator of neoplasia, metastasis, and cytokine suppression. Human genome sequencing has discovered more than 500 kinases. Mutations of the kinase itself or the pathway regulated by kinases leads to the progression of diseases such as Alzheimer's, viral infections, and cancers. Cancer chemotherapy has made significant leaps in recent years. The utilization of chemotherapeutic agents for treating cancers has become difficult due to their unpredictable nature and their toxicity toward the host cells. Therefore, targeted therapy as a therapeutic option against cancer-specific cells and toward the signaling pathways is a valuable avenue of research. SARS-CoV-2 is a member of the Betacoronavirus genus that is responsible for causing the COVID pandemic. Kinase family provides a valuable source of biological targets against cancers and for recent COVID infections. Kinases such as tyrosine kinases, Rho kinase, Bruton tyrosine kinase, ABL kinases, and NAK kinases play an important role in the modulation of signaling pathways involved in both cancers and viral infections such as COVID. These kinase inhibitors consist of multiple protein targets such as the viral replication machinery and specific molecules targeting signaling pathways for cancer. Thus, kinase inhibitors can be used for their anti-inflammatory, anti-fibrotic activity along with cytokine suppression in cases of COVID. The main goal of this review is to focus on the pharmacology of kinase inhibitors for cancer and COVID, as well as ideas for future development.

Computational exploration of molecular flexibility and interaction of meropenem analogs with the active site of oxacillinase-23 in Acinetobacter baumannii.

Background: Carbapenem-resistant Acinetobacter baumannii is an opportunistic pathogen responsible for nosocomial infections and is one of the biggest global threats according to the World Health Organization (WHO), particularly causing substantial morbidity and mortality. Objectives: This study aimed at using computational approaches to screen meropenem and its analogs against OXA-23-positive Acinetobacter baumannii, analyzing the correlations between kinetic and phenotypic characteristics. Methods: A total of 5,450 compounds were screened using virtual screening workflow (HTVS, Glide-SP, and Glide-XP) to identify the best compounds based on their binding energy and interactions against OXA-23 and OXA-27 as they had phenotypic data available. Molecular dynamics simulation and density functional theory (DFT) studies were performed from the outcome of molecular docking analysis. Results: During simulations, meropenem and its analogs exhibited high-level stable interactions with Ser79, Ser126, Thr217, Trp219, and Arg259 of OXA-23. Meropenem displayed a CovDock energy of about -3.5 and -1.9 kcal mol-1 against OXA-23 and OXA-27, respectively. Among the 5,450 compounds, Pubchem_10645796, Pubchem_25224737, and ChEMBL_14 recorded CovDock energy between -6.0 and -9.0 kcal mol-1. Moreover, the infra-red (IR) spectrophotometric analysis revealed C=O and C-N atoms showing bands at 1,800 and 1,125 cm-1, respectively. These observed data are in congruence with the experimental observations. Conclusion: The identified compounds showed good agreement with the spectrophotometric analysis using DFT methods. In the earlier studies, meropenem's MIC value was 32 µg mL-1 in OXA-23-positive isolate A2265 compared to the MIC of 1 µg mL-1 in Δbla OXA-23 A2265. Comparing the CovDock energy and hydrogen-bonding interactions, the predicted results are in good agreement with the experimental data reported earlier. Our results highlight the importance of OXA-23 molecular docking studies and their compliance with the phenotypic results. It will help further in developing newer antibiotics for treating severe infections associated with carbapenem-resistant A. baumannii.

Is AMR in Dairy Products a Threat to Human Health? An Updated Review on the Origin, Prevention, Treatment, and Economic Impacts of Subclinical Mastitis.

Background: Bovine mastitis is the most frequent and costly illness impacting dairy herds worldwide. The presence of subclinical mastitis in dairy cows has an impact on the decreased output of milk and milk quality, culling of affected cows, mortality rate, as well as mastitis-related treatment expenses, generating significant financial loss to the dairy industry. The pathogenic bacteria invade through the mammary gland, which then multiply in the milk-producing tissues causing infection, and the presence of pathogenic bacteria in milk is concerning, jeopardizes human health, and also has public health consequences. Intervention to promote herd health is essential to protect public health and the economy. Results: This review attempts to provide an overview of subclinical mastitis, including mastitis in different species, the effect of mastitis on human health and its pathogenic mechanism, the prevalence and incidence of subclinical mastitis, and current preventive, diagnostic, and treatment methods for subclinical mastitis. It also elaborates on the management practices that should be followed by the farms to improve herd immunity and health. Conclusion: This review brings the importance of the threat of antimicrobial resistance organisms to the dairy industry. Furthermore, this review gives a glimpse of the economic consequences faced by the farmers and a futuristic mastitis market analysis in the dairy industry.

Nimbin analog N2 alleviates high testosterone induced oxidative stress in CHO cells and alters the expression of Tox3 and Dennd1a signal transduction pathway involved in the PCOS zebrafish.

Polycystic ovarian syndrome (PCOS) is a hormonal disorder that causes enlargement of ovaries and follicular maturation arrest, which lacks efficient treatment. N2, a semi-natural triterpenoid from the neem family, was already reported to have antioxidant and antiinflammatory properties in our previous report. This study investigated the anti-androgenic property of N2 on testosterone-induced oxidative stress in Chinese Hamster Ovarian cells (CHO) and PCOS zebrafish model. The testosterone exposure disrupted the antioxidant enzymes and ROS level and enhanced the apoptosis in both CHO cells and PCOS zebrafish. However, N2 significantly protected the CHO cells from ROS and apoptosis. N2 improved the Gonado somatic index (GSI) and upregulated the expression of the SOD enzyme in zebrafish ovaries. Moreover, the testosterone-induced follicular maturation arrest was normalized by N2 treatment in histopathology studies. In addition, the gene expression studies of Tox3 and Denndla in zebrafish demonstrated that N2 could impair PCOS condition. Furthermore, to confirm the N2 activity, the in-silico studies were performed against PCOS susceptible genes Tox3 and Dennd1a using molecular docking and molecular dynamic simulations. The results suggested that N2 alleviated the oxidative stress and apoptosis in-vitro and in-vivo and altered the expression of PCOS key genes.

Identify Potential Urine Biomarkers for Bladder Cancer Prognosis Using NGS Data Analysis and Experimental Validation.

Bladder cancer (BC) is one of the most often reported malignancies globally, with a high recurrence rate and associated morbidity and mortality, especially in advanced BC. There has been a surge in the number of molecular targets revealed for BC prognosis and treatment. However, there is still a great need to discover novel biomarkers. Consequently, the current study investigated biomarkers that might indicate the progression of bladder cancer. In this study, bioinformatics analysis was done on a single GEO dataset, and TCGA-BLCA information was connected with differentially expressed genes (DEGs). The levels of mRNA and protein expression were validated using qRT-PCR. According to our findings, CRYAB, ECM1, ALDOB, AOC, GPX3, IGFBP7, AQP2, LASS2, TMEM176A, GALNT1, and LASS2 were highly enriched in cell division, identical protein binding, and developmental process in bladder cancer patients. In addition, among the highly differentiated genes, ECM1, GALNT1, LASS2, and GPX3 showed significant molecular alterations in BC, which are crucial for marker identification. Moreover, the mRNA, CNVs, and protein levels of ECM1, GALNT1, LASS2, and GPX3 were significantly increased in BC patients. Our predictions and analysis studies stated that these four genes act as urine biomarkers and played a crucial role in disease prognosis and the therapeutic process of bladder cancer. Our outcomes showed that these four novel urine biomarkers have the potential to provide innovative diagnostics, early predictions, and disease targets, ultimately improving the BC patient's prognosis.

Anti-Cancer and Anti-Inflammatory Activities of a Short Molecule, PS14 Derived from the Virulent Cellulose Binding Domain of Aphanomyces invadans, on Human Laryngeal Epithelial Cells and an In Vivo Zebrafish Embryo Model.

In this study, the anti-cancer and anti-inflammatory activities of PS14, a short peptide derived from the cellulase binding domain of pathogenic fungus, Aphanomyces invadans, have been evaluated, in vitro and in vivo. Bioinformatics analysis of PS14 revealed the physicochemical properties and the web-based predictions, which indicate that PS14 is non-toxic, and it has the potential to elicit anti-cancer and anti-inflammatory activities. These in silico results were experimentally validated through in vitro (L6 or Hep-2 cells) and in vivo (zebrafish embryo or larvae) models. Experimental results showed that PS14 is non-toxic in L6 cells and the zebrafish embryo, and it elicits an antitumor effect Hep-2 cells and zebrafish embryos. Anticancer activity assays, in terms of MTT, trypan blue and LDH assays, showed a dose-dependent inhibitory effect on cell proliferation. Moreover, in the epithelial cancer cells and zebrafish embryos, the peptide challenge (i) caused significant changes in the cytomorphology and induced apoptosis; (ii) triggered ROS generation; and (iii) showed a significant up-regulation of anti-cancer genes including BAX, Caspase 3, Caspase 9 and down-regulation of Bcl-2, in vitro. The anti-inflammatory activity of PS14 was observed in the cell-free in vitro assays for the inhibition of proteinase and lipoxygenase, and heat-induced hemolysis and hypotonicity-induced hemolysis. Together, this study has identified that PS14 has anti-cancer and anti-inflammatory activities, while being non-toxic, in vitro and in vivo. Future experiments can focus on the clinical or pharmacodynamics aspects of PS14.

A review on algal mediated synthesis of metal and metal oxide nanoparticles and their emerging biomedical potential.

In the recent two decades, there has been a tremendous increase in the biosynthesis of nanomaterials employing live organisms, their components, extracts, or biomolecules as catalysts. Algae has been used majorly for commercial and industrial uses such as food, feed, skin care, medicines, and fertilizers, algae are now being explored to synthesize green nanoparticles (NPs). Indeed, algae are a rich source of bioactive substances, are easy to produce, grow quickly, and are scalable, therefore this trend is growing by the day. The natural material from algae works as a capping and stabilizing factor in the conversion of metal compounds to metal, metal oxides, or bimetallic NPs. The NPs generated by algae might be intracellular or extracellular, depending on the area of the NPs. The aim of the present review, the first of its kind, is to provide readers with essential information about the diversity of algal strains exploited in the booming field of nanobiotechnology and to explore the biomedical applications of NPs biosynthesized from algae which include antimicrobial, antioxidant, anticancer and biocompatibility properties. Furthermore, this study examines the rationale for the algal-mediated creation of metal, metal oxide, and bimetallic NPs from a variety of algae, as well as the characterization of algae-mediated nanomaterial synthesis.

Efficient One-Pot Synthesis of TiO2/ZrO2/SiO2 Ternary Nanocomposites Using Prunus × Yedoensis Leaf Extract for Enhanced Photocatalytic Dye Degradation.

A simple, efficient, and ecofriendly method was employed to synthesize TiO2/ZrO2/SiO2 ternary nanocomposites using Prunus × yedoensis leaf extract (PYLE) that shows improved photocatalytic and antibacterial properties. The characterization of the obtained nanocomposites was done by X-ray powder diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, field-emission scanning electron microscopy (FE-SEM), and energy-dispersive X-ray spectroscopic (EDS) analysis. The synthesized ternary nanocomposites with nanoscale pore diameters were investigated for the elimination of Reactive Red 120 (RR120) dye. The obtained results showed about 96.2% removal of RR120 dye from aqueous solution under sunlight irradiation. Furthermore, it shows promising antibacterial activity against Staphylococcus aureus and Escherichia coli. The improved photocatalytic and antibacterial activity of TiO2/ZrO2/SiO2 may bring unique insights into the production of ternary nanocomposites and their applications in the environment and biomedical field.

Green Synthesis of Endolichenic Fungi Functionalized Silver Nanoparticles: The Role in Antimicrobial, Anti-Cancer, and Mosquitocidal Activities.

Green nanotechnology is currently a very crucial and indispensable technology for handling diverse problems regarding the living planet. The concoction of reactive oxygen species (ROS) and biologically synthesized silver nanoparticles (AgNPs) has opened new insights in cancer therapy. The current investigation caters to the concept of the involvement of a novel eco-friendly avenue to produce AgNPs employing the wild endolichenic fungus Talaromyces funiculosus. The synthesized Talaromyces funiculosus-AgNPs were evaluated with the aid of UV visible spectroscopy, dynamic light scattering (DLS), Fourier infrared spectroscopy (ATR-FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The synthesized Talaromyces funiculosus-AgNPs (TF-AgNPs) exhibited hemo-compatibility as evidenced by a hemolytic assay. Further, they were evaluated for their efficacy against foodborne pathogens Staphylococcus aureus, Streptococcus faecalis, Listeria innocua, and Micrococcus luteus and nosocomial Pseudomonas aeruginosa, Escherichia coli, Vibrio cholerae, and Bacillus subtilis bacterial strains. The synthesized TF-AgNPs displayed cytotoxicity in a dose-dependent manner against MDA-MB-231 breast carcinoma cells and eventually condensed the chromatin material observed through the Hoechst 33342 stain. Subsequent analysis using flow cytometry and fluorescence microscopy provided the inference of a possible role of intracellular ROS (OH-, O-, H2O2, and O2-) radicals in the destruction of mitochondria, DNA machinery, the nucleus, and overall damage of the cellular machinery of breast cancerous cells. The combined effect of predation by the cyclopoid copepod Mesocyclops aspericornis and TF-AgNPS for the larval management of dengue vectors were provided. A promising larval control was evident after the conjunction of both predatory organisms and bio-fabricated nanoparticles. Thus, this study provides a novel, cost-effective, extracellular approach of TF-AgNPs production with hemo-compatible, antioxidant, and antimicrobial efficacy against both human and foodborne pathogens with cytotoxicity (dose dependent) towards MDA-MB-231 breast carcinoma.

Current Treatment Options for COVID-19 Associated Mucormycosis: Present Status and Future Perspectives.

Mucormycosis has become increasingly associated with COVID-19, leading to the use of the term "COVID-19 associated mucormycosis (CAM)". Treatment of CAM is challenging due to factors such as resistance to many antifungals and underlying co-morbidities. India is particularly at risk for this disease due to the large number of patients with COVID-19 carrying comorbidities that predispose them to the development of mucormycosis. Additionally, mucormycosis treatment is complicated due to the atypical symptoms and delayed presentation after the resolution of COVID-19. Since this disease is associated with increased morbidity and mortality, early identification and diagnosis are desirable to initiate a suitable combination of therapies and control the disease. At present, the first-line treatment involves Amphotericin B and surgical debridement. To overcome limitations associated with surgery (invasive, multiple procedures required) and amphotericin B (toxicity, extended duration and limited clinical success), additional therapies can be utilized as adjuncts or alternatives to reduce treatment duration and improve prognosis. This review discusses the challenges associated with treating CAM and the critical aspects for controlling this invasive fungal infection-early diagnosis and initiation of therapy, reversal of risk factors, and adoption of a multipronged treatment strategy. It also details the various therapeutic options (in vitro, in vivo and human case reports) that have been used for the treatment of CAM.