Unlocking the therapeutic potential of bioactive exopolysaccharide produced by marine actinobacterium Streptomyces vinaceusdrappus AMG31: A novel approach to drug development.

Acidic exopolysaccharide (EPS) was produced by a marine actinobacterium Streptomyces vinaceusdrappus strain AMG31 with the highest yield of 10.6 g/l. The synthesized EPS has an average molecular weight of 5.1 × 104 g/mol and contains arabinose, glucose, galacturonic acid (0.5:2:2 M ratio), with 39.77 % uronic acid residues and 18.8 % sulfate detected. EPS exhibited antioxidant activities with 93.8 % DPPH radical scavenging and 344.7 µg/mg total antioxidant capacity. It displayed anti-inflammatory effects by inhibiting 5-LOX and COX-2. Regarding the cytotoxic activity, the IC50 values are 301.6 ± 11.8, 260.8 ± 12.2, 29.4 ± 13.5, 351.3 ± 11.2, 254.1 ± 9.8, and 266.5 ± 10.4 µg/ml for PC-3, HEP-2, MCF-7, HCT-116, A-549, HepG-2 respectively, which indicate that the produced EPS does not have strong cytotoxic activities. Moreover, the EPS showed anti-Alzheimer activity via inhibition of the Butyrylcholinesterase enzyme, with the highest percentage of 84.5 % at 100 µg/ml. Interestingly, the EPS showed superior anti-obesity activity by inhibiting lipase enzyme with a rate of 95.3 % compared to orlistat as a positive control (96.8 %) at a concentration of 1000 µg/ml. Additionally, the produced EPS displayed the highest anti-diabetic properties by inhibiting α-amylase (IC50 31.49 µg/ml) and α-glucosidase (IC50 6.48 µg/ml), suggesting antidiabetic potential analogous to acarbose. EPS exhibited promising antibacterial and antibiofilm activity against a wide range of Gram-positive and Gram-negative pathogenic bacteria.

Association of IL-10-592 C > A /-1082 A > G and the TNFα -308 G > A with susceptibility to COVID-19 and clinical outcomes.

BACKGROUND: Variation in host immune responses to SARS-CoV-2 is regulated by multiple genes involved in innate viral response and cytokine storm emergence like IL-10 and TNFa gene polymorphisms. We hypothesize that IL-10; -592 C > A and - 1082 A > G and TNFa-308 G > A are associated with the risk of SARS-COV2 infections and clinical outcome. METHODS: Genotyping, laboratory and radiological investigations were done to 110 COVID-19 patients and 110 healthy subjects, in Ismailia, Egypt. RESULTS: A significant association between the - 592 A allele, A containing genotypes under all models (p < 0.0001), and TNFa A allele with risk to infection was observed but not with the G allele of the - 1082. The - 592 /-1082 CG and the - 592 /-1082/ -308 CGG haplotypes showed higher odds in COVID-19 patients. Severe lung affection was negatively associated with - 592, while positive association was observed with - 1082. Higher D-dimer levels were strongly associated with the - 1082 GG genotype. Survival outcomes were strongly associated with the GA genotype of TNFa. -308 as well as AGG and AAA haplotypes. CONCLUSION: IL-10 and TNFa polymorphisms should be considered for clinical and epidemiological evaluation of COVID-19 patients.

Proteomics and metabolomics analyses of Streptococcus agalactiae isolates from human and animal sources.

Streptococcus agalactiae (S. agalactiae), group B Streptococcus (GBS), a major cause of infection in a wide variety of diseases, have been compared in different human and animal sources. We aimed to compare the bacterial proteome and metabolome profiles of human and animal S. agalactiae strains to delineate biological interactions relevant to infection. With the innovative advancement in mass spectrometry, a comparative result between both strains provided a solid impression of different responses to the host. For instance, stress-related proteins (Asp23/Gls24 family envelope stress response protein and heat shock protein 70), which play a role in the survival of GBS under extreme environmental conditions or during treatment, are highly expressed in human and animal strains. One human strain contains ꞵ-lactamase (serine hydrolase) and biofilm regulatory protein (lytR), which are important virulence regulators and potential targets for the design of novel antimicrobials. Another human strain contains the aminoglycosides-resistance bifunctional AAC/APH (A0A0U2QMQ5) protein, which confers resistance to almost all clinically used aminoglycosides. Fifteen different metabolites were annotated between the two groups. L-aspartic acid, ureidopropionic acid, adenosine monophosphate, L-tryptophan, and guanosine monophosphate were annotated at higher levels in human strains. Butyric acid, fumaric acid, isoleucine, leucine, and hippuric acid have been found in both human and animal strains. Certain metabolites were uniquely expressed in animal strains, with fold changes greater than 2. For example, putrescine modulates biofilm formation. Overall, this study provides biological insights into the substantial possible bacterial response reflected in its macromolecular production, either at the proteomic or metabolomic level.

Assessment of post-traumatic stress disorder and health-related quality of life among patients recovered from mild-to-moderate COVID-19 and their close contacts: A cross-sectional study.

The pandemic of COVID-19 is a traumatic event with distressing implications for mental health and several aspects of life. This study aimed to assess Post-traumatic Stress Disorder (PTSD) and Health-Related Quality of Life (HRQoL) among patients who experienced mild-to-moderate COVID-19 and their household contacts. This cross-sectional descriptive study was conducted between May 2020 and November 2020, in Ismailia and Suez governorates. A convenient sampling method was used. The sample size was calculated according to the prevalence of PTSD in COVID-19 patients to be 200 adult participants from both sexes, 100 patients with history of recent COVID-19 infection (up to 2 months after recovery) and their close family. COVID-19 was confirmed by a nasal swab sample tested by PCR in addition to suggestive symptoms and/or positive computed tomography lung findings. Data was collected by using the Arabic version of the Post-Traumatic Stress disorder checklist 5 (PCL5-PTSD) and the Arabic version of Health-Related Quality of Life (HRQoL). The questionnaire was collected through interviews at the Endemic and Infectious diseases and Family medicine outpatient clinics. Of the 200 studied participants, evidence of PTSD was found in 112 (56%) participants; all COVID-19 cases and 12 of their close-contact relatives. Impaired HRQoL was evident in 107 (53.5%) participants; all the COVID-19 cases and 7 contacts. The mean values total scores of the PTSD and HRQoL and its domains were significantly higher among cases compared to their contacts. The most frequently impaired domain was social (55%), psychological (54.5%), impairment (53.5%) followed by physical (48.5%) and the least was health perception (33.5%). The mean values of PTSD, HRQoL and their domains were significantly higher among participants who recalled respiratory symptoms and who had comorbid illness. The odds ratio of PTSD and HRQoL was 0.27 (95% CI: 0.2-0.36) and 0.3 (95% CI: 0.23-0.39) in participants who had more than a case of COVID-19 within their families. All the studied COVID-19 participants showed PTSD and impaired HRQoL, compared to 12% and 7% of their contacts, respectively. Past COVID-19 was associated with impairment of all domains of HRQoL and the most affected were the physical, psychological and impairment domains.

Knowledge, attitude, and practice (KAP) of antimicrobial prescription and its resistance among health care providers in the COVID-19 era: A cross sectional study.

Antimicrobial resistance (AMR) is considered as a global health and development threat. During COVID-19 pandemic, there has been an increase in antimicrobial resistance. Health care providers (HCPs) play the main role in facing antibiotic resistance because they have the authority to prescribe antibiotics during clinical practice as well as in promoting patients' compliance with therapies and avoid self-medication. So, this study will serve as an important source of information in context with Covid19 pandemic in Egypt. The data was collected using a validated standardized self-administered online questionnaire compromised of four sections: socio-demographic data of the HCPs, the general knowledge on antibiotics and AMR, the HCP attitude towards antibiotic prescription and AMR and the practice in applying the appropriate antibiotic prescription. Most of HCPs (93.7%) recorded good knowledge level about antibiotic prescription and antimicrobial resistance with mean score of knowledge13.21 ± 1.83. About 79% of HCPs recorded a positive attitude towards proper antibiotic prescription with mean score of attitudes 63.02 ± 7.68. Fifty four percent of HCPs demonstrated a good level of practice with mean score of practice 9.75 ± 2.17. In conclusion, HCPs in our study have a good level of knowledge and attitude about antibiotics. However lower level of proper practice towards the problem of AMR in COVID19 era was noticed. Implementation of effective policies and guidelines is crucial to evaluate the antimicrobial use especially in the COVID-19 era to reduce the unintended consequences of the misuse of antibiotics and its impact on AMR.

Expression Signature of Immune-Related MicroRNAs in Autoimmune Skin Disease: Psoriasis and Vitiligo Insights.

BACKGROUND: Psoriasis and vitiligo are both chronic, skin-specific diseases classified as autoimmune diseases due to the involvement of several biochemical pathways in their pathogenesis, similar to those altered in other autoimmune diseases. The role of miRNAs in regulating skin autoimmune function has yet to be fully characterized. AIM: The aim of this study was to assess the expression profile of a panel of 11 circulating immune-related miRNAs in patients with autoimmune skin diseases, specifically psoriasis and vitiligo, and correlate their expression signature with the clinicopathological features of the diseases. SUBJECTS AND METHODS: Relative gene expression quantification for 11 immune-related circulating miRNAs in plasma was done for 300 subjects-100 patients with psoriasis, 100 patients with vitiligo and 100 normal healthy volunteers-followed by different modalities of bioinformatics analysis for the results. RESULTS: The expression levels of all the studied immune-related miRNAs were elevated in both autoimmune skin disorders, with much higher levels of expression in psoriasis than in vitiligo patients. There was a significant correlation between most of the studied miRNAs, suggesting shared target genes and/or pathways. Moreover, all the studied miRNAs showed significant results as biomarkers for autoimmune skin disease, with miRNA-145 being the best candidate. Regarding the clinicopathological data, miRNA-7, miRNA-9, miRNA-145, miRNA-148a, and miRNA-148b were positively correlated with age. All the miRNAs were inversely correlated with obesity and disease duration. CONCLUSION: This study highlights the critical role of miRNAs in skin-specific autoimmune diseases that proved to be potential biomarkers for autoimmune skin disorders, warranting their exploration as therapeutic targets.

Determinants of coronavirus disease 2019 vaccine acceptance, hesitancy, and barriers among healthcare workers in Ismailia, Egypt: a mixed methods study.

BACKGROUND: Coronavirus disease 2019 vaccine acceptance among healthcare workers (HCWs) plays a fundamental role in combating the COVID-19 pandemic. Vaccination acceptance depends on perceptions of competence and motives of the providers, producers, health professionals, and policymakers. This study aimed to identify the frequency and determinants of COVID-19 vaccine uptake acceptance, hesitancy, and barriers among HCWs. METHODS: A total of 500 HCWs from 3 different hospital affiliations in Ismailia Governorate, Egypt, were included in this mixed methods study. The study was conducted between March and April 2021 through two phases. Phase 1 included a cross-sectional study using a self-administered questionnaire with inquiries about different determinants of vaccine acceptance, hesitancy, and barriers, which was completed either electronically in a Google form or a hard printed copy. Phase 2 included four focus group discussions to explore the determinants in depth. RESULTS: The mean age of participants was 33.9 ± 7.9 years, 70% were females, 51.2% were nurses, and 28.6% were physicians. Of the 500 HCWs, only 27.8% accepted taking the vaccine immediately, 49.2% refused, and 23% were hesitant. Vaccine safety was the highest associated factor with vaccine acceptance (odds ratio (OR) = 6.3, 95% confidence interval (CI) 3.132-12.619), followed by previous uptake of influenza vaccine (OR = 3.3, 95% CI 2.048-5.217) and vaccine effectiveness (OR = 2.6, 95% CI 1.282-5.262). The main barriers to vaccine acceptance were mistrust in governmental policies during the pandemic or in the healthcare facility they work in. Hesitancy was common among females and nurses and was attributed to three prominent factors, including safety concerns, negative stories, and personal knowledge. CONCLUSIONS: The vaccine acceptance frequency among HCWs was considered low, as the majority either refused or was hesitant about taking the vaccine. Concerns about vaccine safety and effectiveness were significant determinants of vaccine acceptance. Factors related to trust were the main barriers to vaccine uptake. The health authority should establish a surveillance system for side effects of the COVID-19 vaccine and communicate this information between HCWs to decrease their worries about safety and increase vaccine uptake.

Perception of COVID-19 and Vaccine Acceptance among Healthcare Workers.

Background: COVID-19 infection is more likely to be acquired and transmitted by healthcare workers (HCWs). Furthermore, they serve as role models for communities in terms of COVID-19 vaccination attitudes. As a result, HCWs' reluctance to vaccinate could have a significant impact on pandemic containment efforts. Aim: To characterize the current COVID-19 vaccine approval situation among healthcare workers and to determine the most likely reason for agreement or disagreement with COVID-19 vaccination. Methods: This cross-sectional design included 451 HCWs from COVID-19 treatment institutions, with COVID-19 exposure risk changing depending on job function and working location. Results: The study recruited 156 physicians and 295 nurses, of whom 58.1% were female and 41.9% were male. Physicians had a significantly higher rate of participation in COVID-19 pandemic prevention and control, with a rate of 69.9% versus 55.3% of nurses. Acceptance of COVID-19 vaccination was reported by 40.8% of HCWs. The rate of acceptance was significantly higher among physicians (55.1%) than among nurses (33.2%) (p < 0.001). Most HCWs (67.8%) believed the vaccine was not effective. Physicians showed more significant trust in the effectiveness of the vaccine than nurses (41% and 27.5, respectively) (p=0.003). Concerning vaccine safety, only 32.8% of HCWs believed it was safe. This was significantly higher in physicians (41.7%) than in nurses (28.1%) (p=0.004). Conclusion: Vaccination uncertainty is common among healthcare personnel in Egypt, and this could be a significant barrier to vaccine uptake among the public. Campaigns to raise vaccine knowledge are critically needed.

Evaluation of blood and biochemical parameters of COVID-19 patients in Suez Canal University Hospital; A retrospective study.

INTRODUCTION: The severe acute respiratory syndrome coronavirus 2 (SARS CoV2-CoV-2) viral outbreak in Wuhan (China) caused thousands of confirmed cases and deaths around the world. Severe viral pneumonia with respiratory failure and death are the ultimate consequence of infection. AIM: This study aimed to evaluate the regularly performed standard laboratory parameters that can assist in COVID-19 case identification and establish an effective approach to help care and management of (COVID-19) patients. METHODOLOGY: COVID-19 (n = 129) patients were hospitalized in the Suez Canal University Hospital and were retrospectively examined. Laboratory parameters were gathered from patients upon admission (n = 129) during the period from the 20th of June to 15th of August 2020. SARS-CoV-2 cases were diagnosed clinically and radiologically by chest Computed Tomography (CT) and confirmed by RT-PCR. RESULTS: The results showed that COVID-19 survivors exhibited lower hemoglobin (Hb) and hematocrit (HCT), while showed higher Red Cell Distribution Width (RDW), neutrophil lymphocyte ratio (NLR), and lymphocytes. Logistic regression analysis showed that age greater than 60 years old, neutrophilia and high NLR were associated with more deaths. CONCLUSION: Monitoring of lymphopenia, neutrophilia and NLR may help categorizing patients who may need Intensive care.

Genotypic detection of metronidazole and clarithromycin resistance in dyspeptic patients with helicobacter pylori.

In Egypt, antibiotic sensitivity analysis for Helicobacter pylori is not routinely performed. We aimed to identify the clarithromycin and metronidazole resistance directly from gastric biopsies for better guide treatment regimens. This cross-sectional descriptive study included 75 adult dyspeptic patients referred to the upper endoscopy unit in Suez Canal University Hospital, Ismailia, Egypt. Gastric biopsies were taken for rapid urease test (RUT) and cultured on brucella agar with antibiotic supplements. Genomic DNA was extracted directly from the specimen, and PCR was performed for direct detection of H. pylori. Also, to explore clarithromycin and metronidazole resistance, mutations in the 23S rRNA gene and the rdxA gene were investigated. We found that 60 samples were positive to RUT (80%), and only 4 samples were positive by culture. UreC gene was detected in 45 specimens. Meanwhile, 26 isolates were contained mutations at positions 2142 and 2143. Amplification of the metronidazole rdx gene was performed by conventional PCR. Out of 45 isolates, DNA sequence analysis of PCR product showed the wild type (ACA) in 9 isolates, while the mutant type (ATA) was detected in 28 isolates. We found a significant proportion of clarithromycin and metronidazole resistance among H. pylori infected patients in our region.

Assessment of potential risk factors for coronavirus disease-19 (COVID-19) among health care workers.

BACKGROUND: Understanding COVID-19 infection among health workers and the risk factors for adverse outcomes is important not only for characterizing virus transmission patterns and risk factors for infection, but also for preventing the future infection of health workers and other patients and reducing secondary COVID-19 transmission within health care settings. Our aim was to identify risk factors for infection among health care workers to limit adverse events in health care facilities. SUBJECTS AND METHODS: A total of 336 HCWs from COVID-19 treatment hospitals took part in the study with varying COVID-19 exposure risk depending on job function and working site. All participants were asked about risk factors for COVID-19 infection. RESULTS: Among our participants, 42.6% were medical doctors, 28.6% nurses and 7.4% assistant nurses and 21.4% were others. Forty four percent of participants had work experience 5-10 years. More than half of participants received training in Infection Prevention and Control (IPC) (56.8%) about COVID 19; 91% have hand hygiene facilities and 69% admitted availability of PPE. More than half of participants admitted that they always follow IPC measures. Two thirds of participants (66.7%) had close contact with a patient since admission; 42.3% were present in aerosolizing procedures for patients. Forty two percent of participants had respiratory symptom; the most common was sore throat representing (32.4%). The highest frequency of respiratory symptoms was among of nurses and assistant nurses 51%. Frequency of respiratory symptoms was higher among those who contacted the patient directly or for prolonged period compared to those who do not admitted these contacts. CONCLUSIONS: Risk factors for COVID-19 represented by those who were smokers, nurses and assistant nurses were more liable to catch COVID-19 than doctors as they contacted the patient directly for prolonged period or his/her body fluids, materials or surfaces around him.

Genotyping of Helicobacter pylori Virulence Genes cagA and vacA: Regional and National Study.

Helicobacter pylori (H. pylori) plays a crucial role in the pathogenesis of gastritis, peptic ulcer, and gastric cancer. The presence of pathogenicity islands (PAI) genes contributes to the pathogenesis of many gastrointestinal disorders. Cytotoxin-associated gene A (cagA) and vacuolating cytotoxin gene (vacA) are the most known virulence genes in H. pylori. So, our aim was to study H. pylori virulence genes' role in gastric disorders pathogenesis. Our study included 150 adult patients who suffered dyspeptic symptoms and were referred to the GIT endoscopy unit. Gastric biopsies were attained for rapid urease test (RUT) and histopathological examination, and multiplex PCR technique for detection of virulence genes was performed. It was found that 100 specimens were (RUT) positive, of which sixty samples (60%) were PCR positive for H. pylori ureC gene. The vacA and cagA genes were identified in 61.6% and 53% of H. pylori strains, respectively. Only 5 cases were vacA-positive and cagA-negative. The most virulent vacA s1 allele existed in 56.6% of cases. Out of the 60 H. pylori strains, 66% had at least one virulence gene and 34% did not show any virulence gene. H. pylori infection showed significant increase with age. H. pylori are prevalent amid dyspeptic patients in our region. The main genotype combinations were vacA+/cagA+ of s1m1 genotype and they were frequently associated with peptic ulcer diseases, gastritis, and gastroesophageal reflux disease.

Molecular Insights into Human Transmembrane Protease Serine-2 (TMPS2) Inhibitors against SARS-CoV2: Homology Modelling, Molecular Dynamics, and Docking Studies.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV2), which caused novel corona virus disease-2019 (COVID-19) pandemic, necessitated a global demand for studies related to genes and enzymes of SARS-CoV2. SARS-CoV2 infection depends on the host cell Angiotensin-Converting Enzyme-2 (ACE2) and Transmembrane Serine Protease-2 (TMPRSS2), where the virus uses ACE2 for entry and TMPRSS2 for S protein priming. The TMPRSS2 gene encodes a Transmembrane Protease Serine-2 protein (TMPS2) that belongs to the serine protease family. There is no crystal structure available for TMPS2, therefore, a homology model was required to establish a putative 3D structure for the enzyme. A homology model was constructed using SWISS-MODEL and evaluations were performed through Ramachandran plots, Verify 3D and Protein Statistical Analysis (ProSA). Molecular dynamics simulations were employed to investigate the stability of the constructed model. Docking of TMPS2 inhibitors, camostat, nafamostat, gabexate, and sivelestat, using Molecular Operating Environment (MOE) software, into the constructed model was performed and the protein-ligand complexes were subjected to MD simulations and computational binding affinity calculations. These in silico studies determined the tertiary structure of TMPS2 amino acid sequence and predicted how ligands bind to the model, which is important for drug development for the prevention and treatment of COVID-19.

Efflux MexAB-Mediated Resistance in P. aeruginosa Isolated from Patients with Healthcare Associated Infections.

Today, one of the most important challenges for physicians is the adequate treatment of infections due to multidrug resistant organism (MDR). Pseudomonas aeruginosa is considered an opportunistic organism causing different types of healthcare associated infections (HAIs). We aimed to investigate the MDR and pandrug resistance (PDR) rate in P. aeruginosa in our region and detect efflux-pump mexAB genes and the proposed binding interactions of five different categories of antimicrobial agents with the mexB pump. A total of 180 non-duplicated P. aeruginosa strains were isolated from patients with HAIs in the Suez Canal University Hospital. Phenotypically, minimum inhibitory concentration (MIC) was done for all MDR and PDR strains before and after addition of efflux pump inhibitor carbonyl cyanide m-chlorophenyl hydrazone (CCCP). Molecular detection of mexA and mexB genes was done by using polymerase chain reaction (PCR). Most of the isolated strains (126 strains) were MDR (70%); only 10 samples (5.5%) were PDR. MexA and mexB genes were detected in 88.2% (120 strains) and 70.5% (96 strains) of stains, respectively. All PDR strains (10 stains) carried both mexA and mexB genes. Efflux mexAB genes were detected in all MDR and PDR strains (136 strains). Molecular modeling studies were performed to investigate the modes of intermolecular binding interactions between the antimicrobial agents and mexB key amino acids that resulted in MDR and PDR. The current study reported high prevalence of MDR and PDR P. aeruginosa in patients with HAIs in the Suez Canal University Hospitals.

Strong prediction of virological response to combination therapy by IL28B gene variants rs12979860 and rs8099917 in chronic hepatitis C genotype 4.

BACKGROUND: A strong association between single nucleotide polymorphisms (SNPs) of IL28B and treatment outcomes of pegylated interferon-α (PEG IFNα) and ribavirin (RBV) has been shown in chronic hepatitis C (CHC) patients with genotype 1. AIM: This study aimed to assess two SNPs of IL28B, rs12979860 and rs8099917, in predicting sustained virological responses (SVR) to treatment of CHC patients with genotype 4 (HCV-4). The value of rs8099917 was investigated in carriers of unfavourable genotypes of rs12979860. METHODS: This study included 119 CHC patients with HCV-4 receiving combination therapy. Both SNPs of IL28B were determined by real-time detection polymerase chain reaction. RESULTS: Genotypes CC/CT/TT of rs12979860 were found in 42 (35.3%), 56 (47.1%) and 21 (17.6%) and rs8099917 TT/TG/GG were found in 74 (62.2%), 40 (33.6%) and 5 (4.2%). In carriers of rs12979860 CC and rs8099917 TT, the rate of SVR was 87.5 and 65.7% respectively. In 54 patients heterozygous for the C allele of rs12979860, testing of rs8099917 revealed SVR in 42.3% of carriers of the TT genotype but no such responses in carriers of TG or GG (P < 0.0001, OR = 47.3, 95% CI: 2.33-767.2). By multivariate analysis, predictors of SVR were baseline ALT (P = 0.014, OR = 6.3, 95% CI: 1.45-27.33), rs12979860 CC (P = 0.001, OR = 13.48, 95% CI: 2.95-61.69) and rs8099917 TT (P = 0.027, OR = 7.5, 95% CI: 1.25-44.88). CONCLUSION: In CHC genotype 4 patients, favourable genotypes of both SNPs of IL28B are valuable for predicting SVR. Additional genotyping of rs8099917 in carriers of the heterozygous C allele of rs12979860 can improve the prediction of SVR.