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1.
Heliyon ; 10(7): e28043, 2024 Apr 15.
Article En | MEDLINE | ID: mdl-38586322

Rice, a primary food source in many countries of the world accumulate potentially harmful elements which pose a significant health hazard to consumers. The current study aimed to evaluate potentially toxic and mineral elements in both paddy soils and rice grains associated with allied health risks in Malakand, Pakistan. Rice plants with intact root soil were randomly collected from paddy fields and analyzed for mineral and potentially toxic elements (PTEs) through inductively coupled plasma optical emission spectrometry (ICP‒OES). Through deterministic and probabilistic risk assessment models, the daily intake of PTEs with allied health risks from consumption of rice were estimated for children and adults. The results of soil pH (< 8.5) and electrical conductivity (EC > 400 µs/cm), indicated slightly saline nature. The mean phosphorus concentration of 291.50 (mg/kg) in soil samples exceeded FAO/WHO permissible limits. The normalized variation matrix of soil pH with respect to Ni (0.05), Ca (0.05), EC (0.08), and Mg (0.09), indicated significant influence of pH on PTEs mobility. In rice grains, the mean concentrations (mg/kg) of Mg (463.81), Al (70.40), As (1.23), Cr (12.53), Cu (36.07), Fe (144.32), Mn (13.89), and Ni (1.60) exceeded FAO/WHO safety limits. The transfer factor >1 for K, Cu, P and Zn indicated bioavailability and transfer of these elements from soil to rice grains. Monte Carlo simulations of hazard index >1 for Cr, Zn, As, and Cu with certainties of 89.93% and 90.17%, indicated significant noncarcinogenic risks for children and adults from rice consumption. The total carcinogenic risk (TCR) for adults and children exceeded the USEPA acceptable limits of 1×10-6 to 1×10-4, respectively. The sensitivity analysis showed that the ingestion rate was a key risk factor. Arsenic (As) primarily influenced total cancer risk (TCR) in children, while chromium (Cr) significantly impacted adults. Deterministic cancer risk values slightly exceeded probabilistic values due to inherent uncertainties in deterministic analysis. Rice consumption poses health risks, mainly from exposure to Cr, Ni and As in the investigated area.

4.
PLoS One ; 19(2): e0297367, 2024.
Article En | MEDLINE | ID: mdl-38394191

Single nucleotide polymorphisms are the most common form of DNA alterations at the level of a single nucleotide in the genomic sequence. Genome-wide association studies (GWAS) were carried to identify potential risk genes or genomic regions by screening for SNPs associated with disease. Recent studies have shown that SCN9A comprises the NaV1.7 subunit, Na+ channels have a gene encoding of 1988 amino acids arranged into 4 domains, all with 6 transmembrane regions, and are mainly found in dorsal root ganglion (DRG) neurons and sympathetic ganglion neurons. Multiple forms of acute hypersensitivity conditions, such as primary erythermalgia, congenital analgesia, and paroxysmal pain syndrome have been linked to polymorphisms in the SCN9A gene. Under this study, we utilized a variety of computational tools to explore out nsSNPs that are potentially damaging to heath by modifying the structure or activity of the SCN9A protein. Over 14 potentially damaging and disease-causing nsSNPs (E1889D, L1802P, F1782V, D1778N, C1370Y, V1311M, Y1248H, F1237L, M936V, I929T, V877E, D743Y, C710W, D623H) were identified by a variety of algorithms, including SNPnexus, SNAP-2, PANTHER, PhD-SNP, SNP & GO, I-Mutant, and ConSurf. Homology modeling, structure validation, and protein-ligand interactions also were performed to confirm 5 notable substitutions (L1802P, F1782V, D1778N, V1311M, and M936V). Such nsSNPs may become the center of further studies into a variety of disorders brought by SCN9A dysfunction. Using in-silico strategies for assessing SCN9A genetic variations will aid in organizing large-scale investigations and developing targeted therapeutics for disorders linked to these variations.


Genome-Wide Association Study , Polymorphism, Single Nucleotide , Humans , Molecular Dynamics Simulation , Mutation , Algorithms , NAV1.7 Voltage-Gated Sodium Channel/genetics
5.
Curr Top Med Chem ; 23(30): 2844-2862, 2023.
Article En | MEDLINE | ID: mdl-38031798

Cancer is considered one of the deadliest diseases globally, and continuous research is being carried out to find novel potential therapies for myriad cancer types that affect the human body. Researchers are hunting for innovative remedies to minimize the toxic effects of conventional therapies being driven by cancer, which is emerging as pivotal causes of mortality worldwide. Cancer progression steers the formation of heterogeneous behavior, including self-sustaining proliferation, malignancy, and evasion of apoptosis, tissue invasion, and metastasis of cells inside the tumor with distinct molecular features. The complexity of cancer therapeutics demands advanced approaches to comprehend the underlying mechanisms and potential therapies. Precision medicine and cancer therapies both rely on drug discovery. In vitro drug screening and in vivo animal trials are the mainstays of traditional approaches for drug development; however, both techniques are laborious and expensive. Omics data explosion in the last decade has made it possible to discover efficient anti-cancer drugs via computational drug discovery approaches. Computational techniques such as computer-aided drug design have become an essential drug discovery tool and a keystone for novel drug development methods. In this review, we seek to provide an overview of computational drug discovery procedures comprising the target sites prediction, drug discovery based on structure and ligand-based design, quantitative structure-activity relationship (QSAR), molecular docking calculations, and molecular dynamics simulations with a focus on cancer therapeutics. The applications of artificial intelligence, databases, and computational tools in drug discovery procedures, as well as successfully computationally designed drugs, have been discussed to highlight the significance and recent trends in drug discovery against cancer. The current review describes the advanced computer-aided drug design methods that would be helpful in the designing of novel cancer therapies.


Antineoplastic Agents , Neoplasms , Animals , Humans , Molecular Docking Simulation , Computer-Aided Design , Artificial Intelligence , Drug Design , Drug Discovery , Neoplasms/drug therapy , Antineoplastic Agents/chemistry
6.
PLoS One ; 18(8): e0289900, 2023.
Article En | MEDLINE | ID: mdl-37590216

There is now widespread agreement that global warming is the source of climate variability and is a global danger that poses a significant challenge for the 21st century. Climate crisis has exacerbated water deficit stress and restricts plant's growth and output by limiting nutrient absorption and raising osmotic strains. Worldwide, Sweet pepper is among the most important vegetable crops due to its medicinal and nutritional benefits. Drought stress poses negative impacts on sweet pepper (Capsicum annuum L.) growth and production. Although, γ aminobutyric acid (GABA) being an endogenous signaling molecule and metabolite has high physio-molecular activity in plant's cells and could induce tolerance to water stress regimes, but little is known about its influence on sweet pepper development when applied exogenously. The current study sought to comprehend the effects of foliar GABA application on vegetative development, as well as physiological and biochemical constituents of Capsicum annuum L. A Field experiment was carried out during the 2021 pepper growing season and GABA (0, 2, and 4mM) concentrated solutions were sprayed on two Capsicum annuum L. genotypes including Scope F1 and Mercury, under drought stress of 50% and 30% field capacity. Results of the study showed that exogenous GABA supplementation significantly improved vegetative growth attributes such as, shoot and root length, fresh and dry weight, as well as root shoot ratio (RSR), and relative water content (RWC) while decreasing electrolyte leakage (EL). Furthermore, a positive and significant effect on chlorophyll a, b, a/b ratio and total chlorophyll content (TCC), carotenoids content (CC), soluble protein content (SPC), soluble sugars content (SSC), total proline content (TPC), catalase (CAT), and ascorbate peroxidase (APX) activity was observed. The application of GABA at 2mM yielded the highest values for these variables. In both genotypes, peroxidase (POD) and superoxide dismutase (SOD) content increased with growing activity of those antioxidant enzymes in treated plants compared to non-treated plants. In comparison with the rest of GABA treatments, 2mM GABA solution had the highest improvement in morphological traits, and biochemical composition. In conclusion, GABA application can improve development and productivity of Capsicum annuum L. under drought stress regimes. In addition, foliar applied GABA ameliorated the levels of osmolytes and the activities of antioxidant enzymes involved in defense mechanism.


Capsicum , Antioxidants , Chlorophyll A , Droughts , Crops, Agricultural , Dehydration
7.
PLoS One ; 18(4): e0284993, 2023.
Article En | MEDLINE | ID: mdl-37099543

Nitric Oxide (NO) signaling pathway plays a vital role in various physiological and pathophysiological processes including vasodilation, neurogenesis, inflammation, translation and protein regulation. NO signaling pathway is associated with various diseases such as cardiovascular diseases, vision impairment, hypertension and Alzheimer's disease. Human Endothelial Nitric Oxide Synthase (eNOS) bound with calcium regulatory protein (calmodulin (CaM)) to produce NO which initiates cGMP pathway. The current study employs to screen the novel compounds against human eNOS independent of calcium regulatory protein (CaM). The current effort emphasized that the deficiency of CaM leads to dysfunction of cGMP signaling pathway. In this work, a hybrid approach of high-throughput virtual screening and comparative molecular docking studies followed by molecular dynamic simulation analyses were applied. The screening of top ranked two novel compounds against eNOS were reported that showed effective binding affinity, retrieved through the DrugBank and ZINC database libraries. Comparative molecular docking analyses revealed that Val-104, Phe-105, Gln-247, Arg-250, Ala-266, Trp-330, Tyr-331, Pro-334, Ala-335, Val-336, Tyr-357, Met-358, Thr-360, Glu-361, Ile-362, Arg-365, Asn-366, Asp-369, Arg-372, Trp-447 and Tyr-475 are potent residues for interactional studies. High-throughput virtual screening approach coupled with molecular dynamic simulation and drug likeness rules depicted that ZINC59677432 and DB00456 are potent compounds to target eNOS. In conclusion, the proposed compounds are potent against eNOS based on extensive in silico analyses. Overall, the findings of this study may be helpful to design therapeutic targets against eNOS.


Calcium , Nitric Oxide Synthase Type III , Humans , Trypsin , Molecular Docking Simulation , Amino Acid Sequence , Calmodulin , Peptide Fragments
8.
Molecules ; 28(7)2023 Apr 05.
Article En | MEDLINE | ID: mdl-37050015

Titanium dioxide (TiO2) has been investigated for solar-energy-driven photoelectrical water splitting due to its suitable band gap, abundance, cost savings, environmental friendliness, and chemical stability. However, its poor conductivity, weak light absorption, and large indirect bandgap (3.2 eV) has limited its application in water splitting. In this study, we precisely targeted these limitations using first-principle techniques. TiO2 only absorbs near-ultraviolet radiation; therefore, the substitution (2.1%) of Ag, Fe, and Co in TiO2 significantly altered its physical properties and shifted the bandgap from the ultraviolet to the visible region. Cobalt (Co) substitution in TiO2 resulted in high absorption and photoconductivity and a low bandgap energy suitable for the reduction in water without the need for external energy. The calculated elastic properties of Co-doped TiO2 indicate the ductile nature of the material with a strong average bond strength. Co-doped TiO2 exhibited fewer microcracks with a mechanically stable composition.

9.
Fish Physiol Biochem ; 49(2): 275-287, 2023 Apr.
Article En | MEDLINE | ID: mdl-36920633

This study tested the reverence of background color with growth, movement behavior, and some body physiological factors in Nile tilapia. Fish were first acclimatized for two weeks in 80 L glass aquaria. In the experimental design, three groups were maintained separately. In 1st group, glass aquaria were covered with black, 2nd with green charts, and 3rd remained uncovered and maintained as a control group. All groups were in three replicates and fed on a 30% protein diet. All fishes were closely observed for movement behavior and growth-related parameters. On the 28th day, fish were anesthetized and blood was drawn from the caudal fin to determine some hematological parameters, cortisol, and glucose level. The highest weight gain and the greatest number of movements throughout the aquarium were observed in the control group. Furthermore, aquarium color determined the body pigmentation color. Black aquarium had the lowest weight gain, glucose level, FCR, DFI, FCE, and CF, while green aquarium had the highest cortisol level. Histological changes observed in black aquaria fish included reduced lamellar size, deformed blood vessels, and cardiac muscle fascicle in the gills, liver, and heart, respectively, while green aquaria fish showed fused lamellae, necrosis, fibrosis, endomiocardiac hyperplasia, and cardiac muscle fascicle. It was concluded that background coloration has a significant effect on the growth performance, behavior, hematology, body color, histology, cortisol, and glucose levels of Oreochromis niloticus, with green background inducing significant stress while black background causes black pigmentation with less growth.


Cichlids , Animals , Hydrocortisone , Body Weight , Weight Gain , Glucose , Animal Feed/analysis , Diet , Dietary Supplements
10.
Curr Neuropharmacol ; 21(5): 1026-1041, 2023.
Article En | MEDLINE | ID: mdl-36918785

With the advancement in novel drug discovery, biologically active compounds are considered pharmacological tools to understand complex biological mechanisms and the identification of potent therapeutic agents. Mitochondria boast a central role in different integral biological processes and mitochondrial dysfunction is associated with multiple pathologies. It is, therefore, prudent to target mitochondrial quality control mechanisms by using pharmacological approaches. However, there is a scarcity of biologically active molecules, which can interact with mitochondria directly. Currently, the chemical compounds used to induce mitophagy include oligomycin and antimycin A for impaired respiration and acute dissipation of mitochondrial membrane potential by using CCCP/FCCP, the mitochondrial uncouplers. These chemical probes alter the homeostasis of the mitochondria and limit our understanding of the energy regulatory mechanisms. Efforts are underway to find molecules that can bring about selective removal of defective mitochondria without compromising normal mitochondrial respiration. In this report, we have tried to summarize and status of the recently reported modulators of mitophagy.


Mitochondria , Mitophagy , Humans , Mitophagy/physiology , Mitochondria/metabolism , Membrane Potential, Mitochondrial , Antimycin A/metabolism
11.
Environ Health ; 21(1): 130, 2022 12 15.
Article En | MEDLINE | ID: mdl-36517903

AIM: To explore the health risk of living near permitted composting sites (PCSs) on disease severity in children and adults with cystic fibrosis (CF) across the UK.  METHODS: A semi-individual cross-sectional study was used to examine the risk of disease severity in people with CF (pwCF) within and beyond 4 km of PCSs in the UK in 2016. All pwCF registered in the UK CF Registry were eligible for this study. Linear and Poisson regressions, adjusted for age, gender, genotype, BMI, Pseudomonas aeruginosa and deprivation, were used to quantify associations between distance to a PCS and percent predicted forced expiratory volume in one second (ppFEV1), pulmonary exacerbations (#IVdays), and fungal and bacterial infections. RESULTS: The mean age of the 9,361 pwCF (3,931 children and 5,430 adults) studied was 20.1 (SD = 14.1) years; 53.3% were male; and 49.2% were homozygous F508del. Over 10% of pwCF (n = 1,015) lived within 4 km of a PCS. We found no statistically significant difference in ppFEV1 and #IVdays/year in children. However, in adults, ppFEV1 was -1.07% lower (95% confidence interval (CI): -2.29%, 0.16%) and #IVdays/year were 1.02 day higher (95%CI: 1.01, 1.04) within 4 km of a PCS. Furthermore, there were statistically significant differences in mean ppFEV1 in CF adults with Aspergillus fumigatus (58.2.% vs 62.0%, p = 0.005) and Candida spp. (56.9% vs 59.9%, p = 0.029) residing within 4 km of a PCS. No associations were identified for allergic bronchopulmonary aspergillosis, P. aeruginosa or Staphylococcus aureus. CONCLUSIONS: This novel national study provides evidence that adults with CF living near a PCS may experience small reductions in lung function, an increased risk of pulmonary exacerbations, and more frequent fungal infections. If confirmed by studies using refined exposure assessment methods accounting for bioaerosol dispersion, these results could have important implications for the living environment of pwCF.


Bacterial Infections , Composting , Cystic Fibrosis , Lung , Adult , Child , Female , Humans , Male , Young Adult , Cross-Sectional Studies , Cystic Fibrosis/epidemiology , Cystic Fibrosis/genetics , Cystic Fibrosis/microbiology , Lung/physiopathology , Registries , United Kingdom/epidemiology
12.
PLoS One ; 17(12): e0279083, 2022.
Article En | MEDLINE | ID: mdl-36542623

Water is a vital, finite resource whose quantity and quality are deteriorating as the world population increases. The current study aims to investigate the concentration of heavy metals (HM) in surface water for irrigation purposes with associated human health risks and pollution sources near the marble industry in Malakand, Pakistan. Twenty-seven water samples were randomly collected and analyzed for HM concentration by inductively coupled plasma‒optical emission spectrometry (ICP‒OES). pH, electrical conductivity (EC), total dissolved solids (TDS), biological oxygen demand (BOD), and chemical oxygen demand (COD) were measured using standard methods of American Public Health Association (APHA). Irrigation suitability was assessed using specific water quality parameters. The associated health risks from ingestion and dermal exposure to heavy metals were calculated by USEPA health risk indices. Pollution sources and spatial distribution mapping were studied using compositional data analysis (CoDa) and the application of a geographic information system (GIS) to understand the changing behavior of heavy metals in surface waters. The concentrations of BOD (89%), COD (89%), Al (89%), Ca (89%), Cr (56%), Cu (78%), Fe (56%), K (34%) Mg (23%), Mn (56%), Na (89%), Ni (56%), P (89%), and Zn (11%) exceeded the safety limits of National Environmental Quality standards (NEQs) of Pakistan. The results of Kelly's ratio (KR) classified surface water as unsuitable for irrigation. The average daily doses (ADD, mg/kg/day) for Al, Cu, Cr, Fe, Mn, Ni, and Zn were higher in children than in adults. The hazard index (HI) for children and adults was above the threshold (HI > 1), indicating a significant risk of non-carcinogenic toxicity. The carcinogenic risk values for Cr and Ni were above the USEPA limit (1 × 10-6 to 1 × 10-4), suggesting a potential carcinogenic risk for the target population. Principal component analysis (PCA), biplot (CLR), and the CoDa-dendrogram allowed for the identification of elemental associations, and their potential source was anthropogenic rather than natural in origin. Regular monitoring and phytoremediation strategies are proposed to safeguard crops and human health.


Metals, Heavy , Water Pollutants, Chemical , Adult , Child , Humans , Environmental Monitoring/methods , Pakistan , Water Pollutants, Chemical/toxicity , Water Pollutants, Chemical/analysis , Metals, Heavy/toxicity , Metals, Heavy/analysis , Water Quality , Risk Assessment
13.
J Adv Vet Anim Res ; 9(2): 290-294, 2022 Jun.
Article En | MEDLINE | ID: mdl-35891655

Objective: The study was conducted to investigate the gut content and record morphometric and hematological parameters in the common hoopoe (Upupa epops). Materials and Methods: Twenty samples of healthy birds (10 from each sex) were collected from different locations in Okara District, Punjab, Pakistan, from September 2020 to March 2021. Birds were captured live for blood samples and morphometric and gut analyses. Results: It was revealed that the concentrations of different hematological parameters were as follows: hemoglobin, 20.03g/dl; red blood cells, 3.28 × 106/µl; white blood cells, 326.67 × 103/µl; hematocrit, 56.47%; MCV, 173.33 FL; MCH, 57.4 pg; MCHC, 57.4 pg; PLT, 8.33/µl; and RDW, 8.33/µl. The percentages of neutrophils, lymphocytes, monocytes, and eosinophils were 84.67%, 11.67%, 2.00%, and 1.67%, respectively. The gut content of the common hoopoe mostly consisted of Coleoptera and Acrididae larvae. However, Lepidoptera, Gryllotalpidae, and sand were also recorded, along with seeds of Salvadora persica. Conclusions: There were no significant differences between male and female U. epops in feeding content, total weight of the gut, or weight of the empty gut. Regarding the morphometric parameters, there was a significant difference in both sexes' wingspan, body length, and body weight. Males were significantly heavier than females.

14.
Sensors (Basel) ; 22(12)2022 Jun 19.
Article En | MEDLINE | ID: mdl-35746414

Cloud Computing (CC) provides a combination of technologies that allows the user to use the most resources in the least amount of time and with the least amount of money. CC semantics play a critical role in ranking heterogeneous data by using the properties of different cloud services and then achieving the optimal cloud service. Regardless of the efforts made to enable simple access to this CC innovation, in the presence of various organizations delivering comparative services at varying cost and execution levels, it is far more difficult to identify the ideal cloud service based on the user's requirements. In this research, we propose a Cloud-Services-Ranking Agent (CSRA) for analyzing cloud services using end-users' feedback, including Platform as a Service (PaaS), Infrastructure as a Service (IaaS), and Software as a Service (SaaS), based on ontology mapping and selecting the optimal service. The proposed CSRA possesses Machine-Learning (ML) techniques for ranking cloud services using parameters such as availability, security, reliability, and cost. Here, the Quality of Web Service (QWS) dataset is used, which has seven major cloud services categories, ranked from 0-6, to extract the required persuasive features through Sequential Minimal Optimization Regression (SMOreg). The classification outcomes through SMOreg are capable and demonstrate a general accuracy of around 98.71% in identifying optimum cloud services through the identified parameters. The main advantage of SMOreg is that the amount of memory required for SMO is linear. The findings show that our improved model in terms of precision outperforms prevailing techniques such as Multilayer Perceptron (MLP) and Linear Regression (LR).


Cloud Computing , Software , Data Collection , Feedback , Reproducibility of Results
15.
PLoS One ; 17(4): e0266739, 2022.
Article En | MEDLINE | ID: mdl-35446890

Sleep is one of the most important functions of the life. The disturbance in sleep or quality of sleep leads to several dysfunctions of the human body. This study aimed to investigate the prevalence of sleep disorders, their possible risk factors and their association with other health problems. The data was collected from the educational community of the Pakistani population. The Insomnia Severity Index (ISI) was used to evaluate the insomnia and the sleep apnea was evaluated through a simple questionnaire method. The blood samples were collected to perform significant blood tests for clinical investigations. Current research revealed that the individuals in the educational community had poor sleep quality. A total of 1998 individuals from the educational community were surveyed, 1584 (79.28%) of whom had a sleep disorders, including insomnia (45.20%) and sleep apnea (34.08%). The measured onset of age for males and females was 30.35 years and 31.07 years respectively. The Clinical investigations showed that the sleep had significant impact on the hematology of the patients. Higher levels of serum uric acid and blood sugar were recorded with a sleep disorder. The individuals of the educational community were using the sleeping pills. The other associated diseases were mild tension, headaches, migraines, depression, diabetes, obesity, and myopia. The use of beverage, bad mood, medical condition, mental stress, disturbed circadian rhythms, workload and extra use of smartphone were major risk factors of sleep disorders. It was concluded that the insomnia was more prevalent than the sleep apnea. Furthermore, life changes events were directly linked with disturbance of sleep. Tension, depression, headaches, and migraine were more associated with sleep disorders than all other health issues.


Migraine Disorders , Sleep Apnea Syndromes , Sleep Initiation and Maintenance Disorders , Sleep Wake Disorders , Adult , Epidemiologic Studies , Female , Headache/complications , Humans , Male , Migraine Disorders/complications , Pakistan/epidemiology , Prevalence , Risk Factors , Sleep Apnea Syndromes/complications , Sleep Initiation and Maintenance Disorders/complications , Sleep Initiation and Maintenance Disorders/epidemiology , Sleep Wake Disorders/complications , Sleep Wake Disorders/epidemiology , Uric Acid
16.
Environ Sci Pollut Res Int ; 29(27): 40724-40733, 2022 Jun.
Article En | MEDLINE | ID: mdl-35083667

Nanoparticles have numerous applications related to human uses. Titanium dioxide nanoparticles (TiO2-NPs) are extensively used in many daily utilities. The small size particles and larger uses in the industry have led them to become a threatening entity to the living organisms. The unchecked use and dumping in the environment poses a significant toxicological risk to the developing mammalian embryo. The present study was conducted to determine the developmental toxicity and teratogenic effects of TiO2-NPs in murine embryos. The TiO2-NPs were introduced intravenously into pregnant mice graded as T1 (0.52 mg/g BW), T2 (0.7 mg/g BW), and T3 (1.05 mg/g BW) along with control with no dose administration T0 (0.00 mg/g BW). Results recorded after 14 days were resorbed fetuses, dropped wrist, hemorrhages, sacral hygromas, and kinked tails. It was concluded that the exposure of TiO2-NPs in mentioned doses from any source may lead to deleterious effects on the development of an embryo.


Metal Nanoparticles , Nanoparticles , Animals , Embryo, Mammalian , Mammals , Metal Nanoparticles/toxicity , Mice , Nanoparticles/toxicity , Titanium/toxicity
17.
Prep Biochem Biotechnol ; 52(3): 318-324, 2022.
Article En | MEDLINE | ID: mdl-34187304

The receptor binding domain (RBD) of SARS-CoV-2 is located in the C-terminal of S1 subunit of the spike (S) protein which is responsible for recognizing and binding to the angiotensin-converting enzyme 2 (ACE2) receptor. The DNA encoding the SARS-CoV-2 RBD was inserted into pET-28a (+) to construct expression plasmid pET-28a (+)/RBD. The desired RBD protein was produced in E. coli Rosetta (DE) and purified by a Ni-NTA column. The recombinant RBD was analyzed by SDS-PAGE and Western blot. The flow cytometry analysis indicated that the recombinant RBD is capable of binding to human ACE2 (hACE2) in the ACE2-overexpressed HEK293A-hACE2 cells. Our results demonstrated that recombinant RBD expressed in E. coli Rosetta (DE) strain has bioactivities and can be used as an antigen for diagnosis and as a tool for the development of novel anti-viral drugs against SASR-CoV-2.


Angiotensin-Converting Enzyme 2/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Spike Glycoprotein, Coronavirus/genetics , Spike Glycoprotein, Coronavirus/metabolism , Angiotensin-Converting Enzyme 2/genetics , Binding Sites , Escherichia coli/genetics , Flow Cytometry , HEK293 Cells , Humans , Plasmids , Protein Engineering , Protein Interaction Domains and Motifs
18.
Egypt Liver J ; 11(1): 17, 2021.
Article En | MEDLINE | ID: mdl-34777867

BACKGROUND: Hepatitis C virus, a silent killer, has infected 71 million people globally. The recombinant viral antigenic proteins might be used in the early diagnosis of HCV infection. The NS3 and NS5A genes of HCV function in HCV replication and influence host cellular factors that are involved in HCV pathogenesis. The current study was designed to select NS3 and NS5A antigenic sites, amplified, cloned, and expressed in order to find out better assays for diagnosis or drug and vaccine development. The antigenic sites within NS3 and NS5A genes were selected and confirmed through sequencing and were cloned. The antigenic recombinant proteins were expressed in bacterial strain E. coli BL21ply*, and the expression was confirmed by western blotting by using gene-specific and vector-specific antibodies. RESULTS: Specific antigenic regions within the NS3 and NS5A genes of the HCV 3A genotype were amplified. PCR results showed 328 bp and 747 bp antigenic regions, respectively. The regions were confirmed by DNA sequencing and cloned into a bacterial expression vector. Expression analysis showed 12 kDa and 28 kDa of NS3 and NS5A antigenic recombinant proteins, respectively. Taken together, these studies will help to analyze the genetic variability within the local HCV isolates as these antigenic recombinant proteins were quite important in the screening of HCV-infected patients. CONCLUSIONS: This study might help to enhance the progress in the treatment of HCV infection through the modeling of HCV non-structural genes (NS3 and NS5A) from local isolate, and it might also present the viral genes as potential therapeutic targets.

19.
Egypt Liver J ; 11(1): 50, 2021.
Article En | MEDLINE | ID: mdl-34777869

BACKGROUND: This study was conducted to check anxiety level and mental stress in the 200 confirmed hepatitis C patients during the COVID-19 pandemic. The Chinese version of the Beck Anxiety Inventory (BAI) score index was used to measure the anxiety level of HCV-positive patients. BAI score index of different demographic factors such as gender, age, occupation, and education of all the sampled population was calculated. RESULTS: The highest BAI score was recorded in people in the age group of 25-45 years (54.5%). Respondents from public sector employees, own businesses, and postgraduates were highly anxious. A significant difference in BAI score was also recorded between male and female respondents as 33.77% of females were at a severe level of anxiety compared to 17.07% of males. Furthermore, quarantined hepatitis C patients had a significantly higher BAI score (39.5) as compared to non-quarantined patients (27.12), and respondents with HCV infection also had a high BAI score of 37.25 compared to healthy individuals (4.1). Most of the respondents were willing to adopt protective measures against COVID-19. CONCLUSION: This study concluded that people with infectious diseases like hepatitis C had high anxiety levels and mental stress in the COVID-19 pandemic and needed psychological aids for better mental health to handle pandemic conditions.

20.
Front Microbiol ; 12: 706700, 2021.
Article En | MEDLINE | ID: mdl-34504479

Phage-host interactions are likely to have the most critical aspect of phage biology. Phages are the most abundant and ubiquitous infectious acellular entities in the biosphere, where their presence remains elusive. Here, the novel Escherichia coli lytic bacteriophage, named MSK, was isolated from the lysed culture of E. coli C (phix174 host). The genome of phage MSK was sequenced, comprising 45,053 bp with 44.8% G + C composition. In total, 73 open reading frames (ORFs) were predicted, out of which 24 showed a close homology with known functional proteins, including one tRNA-arg; however, the other 49 proteins with no proven function in the genome database were called hypothetical. Electron Microscopy and genome characterization have revealed that MSK phage has a rosette-like tail tip. There were, in total, 46 ORFs which were homologous to the Rtp genome. Among these ORFs, the tail fiber protein with a locus tag of MSK_000019 was homologous to Rtp 43 protein, which determines the host specificity. The other protein, MSK_000046, encodes lipoprotein (cor gene); that protein resembles Rtp 45, responsible for preventing adsorption during cell lysis. Thirteen MSK structural proteins were identified by SDS-PAGE analysis. Out of these, 12 were vital structural proteins, and one was a hypothetical protein. Among these, the protein terminase large (MSK_000072) subunit, which may be involved in DNA packaging and proposed packaging strategy of MSK bacteriophage genome, takes place through headful packaging using the pac-sites. Biosafety assessment of highly stable phage MSK genome analysis has revealed that the phage did not possess virulence genes, which indicates proper phage therapy. MSK phage potentially could be used to inhibit the multidrug-resistant bacteria, including AMP, TCN, and Colistin. Further, a comparative genome and lifestyle study of MSK phage confirmed the highest similarity level (87.18% ANI). These findings suggest it to be a new lytic isolated phage species. Finally, Blast and phylogenetic analysis of the large terminase subunit and tail fiber protein put it in Rtp viruses' genus of family Drexlerviridae.

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