The role of blood groups, vaccine type and gender in predicting the severity of side effects among university students receiving COVID-19 vaccines.

On March 11th, 2020, the World Health Organization (WHO) declared the coronavirus disease 2019 (COVID-19) a pandemic. To control the pandemic, billions of vaccine doses have been administered worldwide. Predictors of COVID-19 vaccine-related side effects are inconsistently described in the literature. This study aimed to identify the predictors of side effects' severity after COVID-19 vaccination among young adult students at Taif University (TU) in Saudi Arabia. An online, anonymous questionnaire was used. Descriptive statistics were calculated for numerical and categorical variables. Possible correlations with other characteristics were identified using the chi-square test. The study included 760 young adult participants from TU. Pain at the injection site (54.7%), headache (45.0%), lethargy and fatigue (43.3%), and fever (37.5%) were the most frequently reported COVID-19 vaccine-related side effects after the first dose. The most frequent side effects were reported among the 20-25-year-old age group for all doses of all vaccines. Females experienced remarkably more side effects after the second (p < 0.001) and third doses (p = 0.002). Moreover, ABO blood groups significantly correlated with vaccine-related side effects after the second dose (p = 0.020). The participants' general health status correlated with the side effects after the first and second doses (p < 0.001 and 0.022, respectively). The predictors of COVID-19 vaccine-related side effects in young, vaccinated people were blood group B, female gender, vaccine type, and poor health status.

Chemoprofiling and antimicrobial activity of medicinal herbs used in the treatment of inflammatory bowel disease.

Inflammatory bowel disease (IBD) is a term utilized to illustrate two different chronic disorders of the gastro-intestinal tract i.e., Crohn's disease and ulcerative colitis. The symptoms of IBD are mainly characterized by inflammation, including abdominal pain, chronic diarrhoea, weight loss, shortening of the colon and rectal bleeding. The objective of this study was to evaluate the antimicrobial activity and Gas Chromatography-Mass Spectrometry (GC-MS) analysis of herbs used in the treatment of IBD in Saudi Arabia. Ethanolic extracts of five different herbs from Saudi Arabia namely Pimpinella anisum (Anise), Foeniculum vulgare (Fennel), Matricaria chamomilla (Chamomile), Linum usitatissimum (Linseed), and Punica granatum (Pomegranate) were prepared by Soxhlet extraction. The systemic chemical composition of the extracts was identified by GC-MS with their relative concentrations. The ethanolic extract of P. anisum, F. vulgare, M. chamomilla, L. usitatissimum, and P. granatum showed the presence of 35, 42, 34, 37, and 47 chemical components in these extracts, respectively. The five extracts and an equal mixture of them were examined for their antimicrobial activity by broth dilution method against different organisms. These included Gram-positive (Staphylococcus aureus), Gram-negative (Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis and Pseudomonas aeruginosa) bacteria and one yeast (Candida albicans). P. anisum, F. vulgare, M. chamomilla, L. usitatissimum, P. granatum and the mixture of all five extracts had good activity against E. coli (MIC=3.125, 0.050, 6.25, 0.050 and 0.100 mg/ml, respectively). P. granatum also had a MIC of 3.125 mg/ml against S. aureus. In conclusion. the plants' extracts and an equal mixture of them showed a narrow spectrum of antimicrobial activity against S. aureus, K. pneumoniae, P. mirabilis, P. aeruginosa and C. albicans.

A New EGFR Inhibitor from Ficus benghalensis Exerted Potential Anti-Inflammatory Activity via Akt/PI3K Pathway Inhibition.

Inflammation is a critical defensive mechanism mainly arising due to the production of prostaglandins via cyclooxygenase enzymes. This study aimed to examine the anti-inflammatory activity of fatty acid glucoside (FAG), which is isolated from Ficus benghalensis against lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The cytotoxic activity of the FAG on RAW 264.7 macrophages was evaluated with an MTT assay. The levels of PGE2 and NO and the activity of iNOS, COX-1, and COX-2 enzymes in LPS-stimulated RAW 264.7 cells were evaluated. The gene expression of IL-6, TNF-α, and PGE2 was investigated by qRT-PCR. The expression of epidermal growth factor receptor (EGFR), Akt, and PI3K proteins was examined using Western blotting analysis. Furthermore, molecular docking of the new FAG against EGFR was investigated. A non-cytotoxic concentration of FAG increased NO release and iNOS activity, inhibited COX-1 and COX-2 activities, and reduced PGE2 levels in LPS-stimulated RAW 264.7 cells. It diminished the expression of TNF-α, IL-6, PGE2, EGFR, Akt, and PI3K. Furthermore, the molecular docking study proposed the potential direct binding of FAG with EGFR with a high affinity. This study showed that FAG is a natural EGFR inhibitor, NO-releasing, and COX-inhibiting anti-inflammatory agent via EGFR/Akt/PI3K pathway inhibition.

Role of polymeric materials in preventing COVID-19 infection.

The safety of personal protective equipment (PPE) is very important, and so is the choice of materials used. The ability of electrostatic charges (ESCs) generated from the friction of engineered materials to attract or repel viruses has a significant impact on their applications. This study examined the ESCs generated on the surface of PPE used by healthcare workers to enhance their potential effectiveness in protecting the wearer from viruses. This is a crucial consideration for the newly emerged severe acute respiratory syndrome corona virus 2 (SARS-CoV-2), which has a negative charge. The magnitudes and signs of generated ESCs on the surfaces of the PPE were determined experimentally using an Ultra Stable Surface DC Voltmeter. The high negative ESCs acquired by the polyethylene disposable cap and facemask are expected to repel negatively charged viruses and prevent them from adhering to the outer layer of the material. Also, the choice of polypropylene for facemasks and gowns is excellent because it is an aggressively negatively charged material in the triboelectric series. This property guarantees that facemasks and gowns can repel viruses from the wearer. However, the positive ESCs generated on latex glove surfaces are of great concern because they can attract negatively charged viruses and create a source of infection. In conclusion, it is necessary to ensure that PPE be made of materials whose surfaces develop a negative ESC to repel viruses, as well as to select polyethylene gloves.

Carpachromene Ameliorates Insulin Resistance in HepG2 Cells via Modulating IR/IRS1/PI3k/Akt/GSK3/FoxO1 Pathway.

Insulin resistance contributes to several disorders including type 2 diabetes and cardiovascular diseases. Carpachromene is a natural active compound that inhibits α-glucosidase enzyme. The aim of the present study is to investigate the potential activity of carpachromene on glucose consumption, metabolism and insulin signalling in a HepG2 cells insulin resistant model. A HepG2 insulin resistant cell model (HepG2/IRM) was established. Cell viability assay of HepG2/IRM cells was performed after carpachromene/metformin treatment. Glucose concentration and glycogen content were determined. Western blot analysis of insulin receptor, IRS1, IRS2, PI3k, Akt, GSK3, FoxO1 proteins after carpachromene treatment was performed. Phosphoenolpyruvate carboxykinase (PEPCK) and hexokinase (HK) enzymes activity was also estimated. Viability of HepG2/IRM cells was over 90% after carpachromene treatment at concentrations 6.3, 10, and 20 µg/mL. Treatment of HepG2/IRM cells with carpachromene decreased glucose concentration in a concentration- and time-dependant manner. In addition, carpachromene increased glycogen content of HepG2/IRM cells. Moreover, carpachromene treatment of HepG2/IRM cells significantly increased the expression of phosphorylated/total ratios of IR, IRS1, PI3K, Akt, GSK3, and FoxO1 proteins. Furthermore, PEPCK enzyme activity was significantly decreased, and HK enzyme activity was significantly increased after carpachromene treatment. The present study examined, for the first time, the potential antidiabetic activity of carpachromene on a biochemical and molecular basis. It increased the expression ratio of insulin receptor and IRS1 which further phosphorylated/activated PI3K/Akt pathway and phosphorylated/inhibited GSK3 and FoxO1 proteins. Our findings revealed that carpachromene showed central molecular regulation of glucose metabolism and insulin signalling via IR/IRS1/ PI3K/Akt/GSK3/FoxO1 pathway.

Alternatively activated macrophages; a double-edged sword in allergic asthma.

BACKGROUND: Macrophages are heterogenous phagocytic cells with an important role in the innate immunity. They are, also, significant contributors in the adaptive immune system. Macrophages are the most abundant immune cells in the lung during allergic asthma, which is the most common chronic respiratory disease of both adults and children. Macrophages activated by Th1 cells are known as M1 macrophages while those activated by IL-4 and IL-13 are called alternatively activated macrophages (AAM) or M2 cells. AAM are subdivided into four distinct subtypes (M2a, M2b, M2c and M2d), depending on the nature of inducing agent and the expressed markers. BODY: IL-4 is the major effector cytokine in both alternative activation of macrophages and pathogenesis of asthma. Thus, the role of M2a macrophages in asthma is a major concern. However, this is controversial. Therefore, further studies are required to improve our knowledge about the role of IL-4-induced macrophages in allergic asthma, through precisive elucidation of the roles of specific M2a proteins in the pathogenesis of asthma. In the current review, we try to illustrate the different functions of M2a macrophages (protective and pathogenic roles) in the pathogenesis of asthma, including explanation of how different M2a proteins and markers act during the pathogenesis of allergic asthma. These include surface markers, enzymes, secreted proteins, chemokines, cytokines, signal transduction proteins and transcription factors. CONCLUSIONS: AAM is considered a double-edged sword in allergic asthma. Finally, we recommend further studies that focus on increased selective expression or suppression of protective and pathogenic M2a markers.

RNA protein kinase SNP at -226 C

BACKGROUND: RNA-dependant protein kinase R (PKR) is a primary mediator in the defence mechanism of interferon against viral replication and pathogenesis during Hepatitis C virus (HCV) infection. In the present study, we have examined the role of Single Nucleotide Polymorphisms (SNPs) in the promoter region of PKR and the serum level of the same protein on the outcome of HCV-infected Egyptian patients. PATIENTS AND METHODS: Genomic DNA was extracted from a total of 135 subjects, including 15 healthy controls, 40 HCV spontaneous resolvers (SRs), and 80 patients with chronic HCV infection. PKR genotyping was assessed using DNA sequencing. Finally, serum levels of PKR, TNF-α, INF-γ, and IL-10 were measured using ELISA technique. RESULTS: Serum levels of PKR, TNF-α, and INF-γ showed a significant increase in SRs as compared to chronic HCV patients. On the other hand, serum levels of IL-10 were significantly higher in chronic HCV patients compared to SRs. The present study demonstrated two novel SNPs in the PKR promoter region: at -226 C/T and -141 C/G. The PKR SNP at -226 C < T correlated with HCV-infected patients (genotype 4a) outcome among Egyptians. Our data showed the unique presence of the TT genotype in SRs group (three patients: 7.5%) in PKR -226 C/T. Interestingly, subjects with the TT genotype were more likely to clear their HCV infection than those with the CC genotype. CONCLUSION: Our work provides more detail about PKR gene polymorphism in HCV genotype 4a as a new clinical tool for anticipating HCV-4a infection outcome.

Design and total synthesis of (-)-codonopsinine, (-)-codonopsine and codonopsinine analogues by O-(2-oxopyrrolidin-5-yl)trichloroacetimidate as amidoalkylating agent with improved antimicrobial activity via solid lipid nanoparticle formulations.

A general strategy towards total synthesis of (-)-codonopsinine, (-)-codonopsine and codonopsinine analogues has been developed from (D)-tartaric acid via the intermediate (3S,4R)-1-methyl-2-oxo-5-(2,2,2-trichloroacetamido)pyrrolidinediacetate (7). α-amidoalkylation studies of 7 with electron rich benzene derivative 8a-g as C-nucleophiles afforded (aryl derivatives) 9a-g. The target compounds 1, 2 and 13c-g were readily obtained from 10a-gvia Grignard addition to the homochiral lactam which was produced by deoxygenation using Lewis-acid followed by deacetylation. The synthesized compounds were loaded onto solid lipid nanoparticle formulations (SLNs) prepared by hot emulsification-ultrasonication technique using Compritol as solid lipid and Pluronic f68 as surfactant. SLNs were fully evaluated and the permeation of synthesized compound from SLNs was assayed against non-formulated compounds through dialysis membranes using Franz cell. The data indicated good physical characteristics of the prepared SLNs, sustaining of release profiles and significant improvement of permeation ability when compared to the non-formulated compounds. The antibacterial and antifungal activities of 1, 2 and 13c-g were determined by disc diffusion and microbroth dilution method to determine the minimum inhibitory concentrations (MIC) against seven microorganisms (Staphyloccus aureus, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Acinetobacter baumannii and Candida albicans). The most active compounds against the Gram positive S. aureus were 1, 13C, 13d, and 13g. Also, 13c, 13d, and 13e had antibacterial activity but not 13f against some Gram negative organisms (E. coli, and P. mirabilis). MIC concentrations against P. aeruginosa, and K. pneumoniae were ≥512 µg/ml, while that against A. baumannii was ≥128 µg/ml except for nanoformulae of 13e and 13f that were 16 and 64 µg/ml, respectively. No antifungal activity against Candida albicans was recorded for all compounds and their nanoformulae (MIC > 1024 µg/ml). SLNs were found to decrease the MIC values for some of the compounds with no effect on the antifungal activity. In conclusion, we demonstrated a novel, straight-forward and economical procedure for the total synthesis of (-)-codonopsinine 1, (-)-codonopsine 2 and codonopsinine analogues 13c-g from simple and commercially available starting materials; d-tartaric acid; with antimicrobial activities against Gram positive and Gram-negative organisms that were improved by SLNs formulations.

High spontaneous clearance of symptomatic iatrogenic acute hepatitis C genotype 4 infection.

Acute hepatitis C (AHC) infection resolves spontaneously in 15% to 40% of patients. Factors favoring spontaneous viral clearance remain undefined. In this study, predictors of spontaneous viral clearance in patients with symptomatic AHC were investigated. Epidemiological, clinical, and virologic parameters were also examined. Patients with symptomatic AHC were enrolled and followed up prospectively. The patients were followed up every 2 weeks in the first month and then monthly for the following 5 months, with a follow-up visit 6 months after the last hepatitis C virus (HCV)-RNA negative sample for those who had cleared the virus. Interleukin (IL)-28B.rs12979860 single-nucleotide polymorphism and HCV genotype were tested at baseline. HCV-RNA was tested during each visit. Patients who remained RNA-positive at 24 weeks were treated with pegylated interferon plus ribavirin for 24 weeks. A total of 30 patients, mostly with iatrogenically acquired AHC genotype 4 infections completed 6-months' follow-up, to either spontaneous clearance or start of treatment. The mean age of the patients was 37 ± 13 years. In total, 67% of patients were females, and the mean incubation period was 7.6 ± 3.5 weeks. Viral clearance occurred spontaneously in 19 (63.3%) patients. The average time to clearance was 24.3 ± 9.6 weeks. A total of 11 patients received therapy, and 8 (72.7%) cleared the virus and had a sustained virologic response to the treatment 24 weeks after the therapy. A total of three patients were treatment nonresponders. IL28B.rs12979860 CC genotype, female gender, and viremia level were not associated with self-limiting AHC in this cohort. In conclusion, patients with symptomatic AHC genotype 4 infection caused by an iatrogenic exposure had higher rates of spontaneous resolution than previously reported. Predicting spontaneous viral clearance after iatrogenic AHC exposure was not possible in this population.

Association of Toll-like receptor 3 and Toll-like receptor 9 single-nucleotide polymorphisms with hepatitis C virus persistence among Egyptians.

Toll-like receptors (TLRs) give the innate immune system a considerable specificity for a large range of pathogens. TLR3 detects dsRNA of viruses while TLR9 recognizes bacterial and viral unmethylated CpG motifs. This study examined whether there is a potential association between single-nucleotide polymorphisms (SNPs) in the TLR3.rs3775290 (c.1377C/T), TLR9.rs5743836 (-1237T→C) and TLR9.rs352140 (G2848A) genes and HCV infection among Egyptian patients and healthcare workers (HCWs). We enrolled 546 subjects (409 HCWs and 137 patients) divided into four groups: group 1 included 265 seronegative, aviremic subjects; group 2 included 25 seronegative, viremic subjects; group 3 included 87 subjects with spontaneously resolved HCV infection; and group 4 included 169 chronic HCV patients. All subjects were genotyped for TLR3.rs3775290, TLR9.rs5743836 and TLR9.rs352140 SNPs by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis. TLR3.rs3775290 "CC" genotype was associated with chronic HCV infection, where there was a significantly greater frequency of this genotype among chronic patients when compared to subjects with spontaneously resolved infection (63.9% vs. 51.9%; p = 0.033; OR = 1.639 and 95% CI = 0.94-2.84). However, this SNP did not correlate with the HCV RNA load among the chronic subjects (p > 0.05). There was no significant difference in TLR9.rs5743836 and TLR9.rs352140 genotype distribution between groups (p > 0.05). Lack of association between the three SNPs was found, as the three SNPs are located on two different chromosomes. In conclusion, the TLR3.rs3775290 "CC" genotype was associated with HCV chronicity, while the TLR9 gene may not play a major role in HCV infection.

Topical Eugenol Successfully Treats Experimental Candida albicans-Induced Keratitis.

PURPOSE: To determine the efficacy of eugenol for the treatment of Candida keratitis in an experimental model. METHODS: The in vitro antifungal activity of eugenol and fluconazole was tested against C. albicans strains via the microbroth dilution method. An experimental model of Candida albicans keratitis was used. Rabbits were classified into those that received no treatment (control; group 1) and those that started eugenol treatment immediately (group 2) or after 4 days (group 3) of keratitis induction (n = 12-16 rabbits/group). The 2 treatment groups were assigned to 50 µL of 4 mg/mL eugenol drops hourly for 15 days, while the control group received saline. Corneal penetration of eugenol was measured using HPLC, and corneal toxicity was evaluated clinically and histopathologically. RESULTS: The in vitro minimum inhibitory concentrations of eugenol and fluconazole against C. albicans were 2 and > 0.4 mg/mL, respectively. A 4-mg/mL preparation of eugenol in propylene glycol was the maximum nontoxic dose on rabbit corneas as suggested by clinical and histopathologic findings. At least 75% of all eugenol-treated eyes recovered from keratitis, with improvement in the remaining 25% of the eyes compared to controls. CONCLUSIONS: Eugenol can act as a natural, safe, and effective treatment for fungal keratitis, regardless of whether treatment is started immediately or after 4 days of keratitis induction.

Correlation between hepatitis C viral load and hepatitis C Core antigenaemia among Egyptians.

Hepatitis C virus (HCV) infection is widespread in Egypt. This study compared HCV RNA with HCVcAg for the detection and quantification of viraemia among a sample of Egyptians. Sera from 80 suspected HCVpositive individuals were tested simultaneously for HCV-RNA load using real-time polymerase chain reaction (PCR) and HCVcAg level using ELISA. Of the 80 samples, 25% were HCV-RNA-negative. HCVcAg was detected in all samples: range 0.4-2462 ng/mL, mean 460 (SD 506) ng/mL. The sensitivity and specificity of HCVcAg were 96.7% and 90.9%, respectively. There was a significant correlation between serum HCV-RNA and HCVcAg levels (r = 0.4, P < 0.0001). HCV-RNA remains the gold standard for diagnosis of active HCV infection but HCVcAg can be used where PCR is not available.

Is hepatitis E virus an emerging problem in industrialized countries?

Hepatitis E virus (HEV) is yearly responsible for approximately 20 million infections worldwide. Although most infections occur in developing countries, HEV appears to be an emerging problem in several industrialized countries, where it is mostly associated with either traveling to an HEV endemic area or contact with pigs, which represent a major reservoir of HEV. The major risk groups for HEV infection and its ensuing complications are elderly men, pregnant women, young children, immunocompromised patients, patients with preexisting liver disease, and workers that come into close contact with HEV-infected animals. Whereas HEV mainly causes acute self-limiting infections, chronic infections may occur among immunocompromised patients (e.g., transplant recipients and human immunodeficiency virus [HIV]-infected patients). Accordingly, HEV-HIV coinfection leads to accelerated liver cirrhosis and increased mortality rates compared to HEV infection alone, which is, except during pregnancy, usually associated with only low mortality. In the Western world, the most common genotype (gt) causing HEV infection is gt 3. Ribavirin (RBV) and interferon have been used successfully for treatment of HEV, but this treatment is contraindicated in certain patient groups. Therefore, novel antiviral compounds are highly needed, especially given that viral isolates with RBV resistance have been recently identified. Moreover, eradication of HEV is hampered by long-term environmental persistence of the virus, which represents a continuous source of the virus. In 2011, the first prophylactic HEV vaccine, Hecolin, was approved in China, but it is not yet globally available. In this review, we will discuss the molecular virology of HEV, mode of transmission in industrialized countries, and potential implications for different specific patient populations.

Occult hepatitis C virus infection among Egyptian hemodialysis patients.

Occult hepatitis C virus (HCV) infection (OCI) was reported in an apparently disease-free state in the absence of liver disease, anti-HCV and HCV-RNA in the serum. The existing data examining the clinical significance of OCI and its potential as a source of HCV infection among hemodialysis patients are very limited. We examined the presence of OCI among patients on maintenance hemodialysis at Minia Governorate, Egypt; an HCV endemic country. A total of 81 subjects with negative markers for HCV were enrolled. HCV-RNA was tested in PBMCs by real-time PCR. For the 81 subjects, the average dialysis duration was 32.7 ± 21.7 months and the average ALT level (±SD) was 26 ± 12 U/L while that of AST was 29 ± 16 U/L. Out of the 81 subjects, three (3.7%) were HCV-RNA positive in PBMCs in the absence of serum anti-HCV and HCV-RNA indicating OCI. The viral load of the OCI subjects ranged from 172 to 4150 IU/ml. History of liver disease was positive in one of the three positive patients. These results highlight the potential risk of HCV transmission from patients within hemodialysis units in Egypt. J. Med. Virol. 88:1388-1393, 2016. © 2016 Wiley Periodicals, Inc.

Interleukin-10.rs1800896 and Interleukin-18.rs1946518 gene polymorphisms could not predict the outcome of hepatitis C virus infection in Egyptian patients treated with pegylated interferon plus ribavirin.

A single-nucleotide polymorphism (SNP) in the interleukin (IL)-28B gene was used as a major predictor of the response to treatment in patients with hepatitis C virus (HCV) infection. Data examining the role of IL-10 and IL-18 gene polymorphisms among HCV genotype 4 (G4)-infected Egyptians in response to pegylated interferon (PEG-IFN) plus ribavirin (RBV) therapy are limited. This study investigated the impact of SNP at IL-10.rs1800896 (at position -1082) and IL-18.rs1946518 genes (at position -607) on the response to PEG-IFN/RBV therapy in HCV-infected Egyptians. This study was carried out on 100 HCV patients treated with PEG-IFN plus RBV and 100 healthy controls. The HCV patients included 50 treatment non-responders (NR) and 50 subjects with sustained virologic response (SVR). Genomic DNA from venous blood of subjects was extracted and IL-10.rs1800896 and IL-18.rs1946518 genotypes were determined using allele-specific amplification and SYBR Green real-time PCR. Linkage disequilibrium between the two SNPs was estimated using Haploview software. The frequency of the IL-10.rs1800896 AA, AG and GG genotypes among non-responders were 16 %, 70 % and 14 % while among SVR subjects, the frequency was 34 %, 60 % and 6 %, respectively (p=0.073). On the other hand, the frequency of the IL-18.rs1946518 AA, AC and CC genotypes among non-responders was 14 %, 50 % and 36 %, respectively, while among responders, these frequencies were 28 %, 44 % and 28 %, (p = 0.220). Both markers were in linkage equilibrium (D' = 0.23; r (2) = 0.052). SNPs in the IL-10.rs1800896 and IL-18.rs1946518 genes could not predict the outcome of HCV infection in Egyptians treated with PEG-IFN/RBV.

Association between IL28B rs12979860 single nucleotide polymorphism and the frequency of colonic Treg in chronically HCV-infected patients.

The IL28B gene is associated with spontaneous or treatment-induced HCV viral clearance. However, the mechanism by which the IL28B single nucleotide polymorphism (SNP) affects the extra-hepatic HCV immune responses and its relationship to HCV pathogenesis have not been thoroughly investigated. To examine the mechanism by which IL28B affects HCV clearance. Forty Egyptian patients with chronic HCV infection receiving an Interferon/ribavirin treatment regimen were enrolled into this study. There were two groups: non-responders (NR; n = 20) and sustained virologic responders (SVR; n = 20). The initial plasma HCV viral loads prior to treatment and IL28B genotypes were determined by quantitative RT-PCR and sequencing, respectively. Liver biopsies were examined to determine the inflammatory score and the stage of fibrosis. Colonic regulatory T cell (Treg) frequency was estimated by immunohistochemistry. No significant association between IL28B genotypes and response to therapy was identified, despite an odds ratio of 3.4 to have the TT genotype in NR compared to SVR (95 % confidence interval 0.3-35.3, p = 0.3). Patients with the TT-IL28Brs12979860 genotype (unfavorable genotype) have significantly higher frequencies of colonic Treg compared to the CT (p = 0.04) and CC (p = 0.03) genotypes. The frequency of colonic Treg cells in HCV-infected patients had a strong association with the IL-28B genotype and may have a significant impact on HCV clearance.

Differential distribution of IL28B.rs12979860 single-nucleotide polymorphism among Egyptian healthcare workers with and without a hepatitis C virus-specific cellular immune response.

The CC genotype of the interleukin (IL)-28B.rs12979860 gene has been associated with spontaneous hepatitis C virus (HCV) clearance and treatment response. The distribution and correlation of an IL28B.rs12979860 single-nucleotide polymorphism (SNP) with HCV-specific cell-mediated immune (CMI) responses among Egyptian healthcare workers (HCWs) is not known. We determined this relationship in 402 HCWs who serve a patient cohort with ~85% HCV prevalence. We enrolled 402 HCWs in four groups: group 1 (n = 258), seronegative aviremic subjects; group 2 (n = 25), seronegative viremic subjects; group 3 (n = 41), subjects with spontaneously resolved HCV infection; and group 4 (n = 78), chronic HCV patients. All subjects were tested for an HCV-specific CMI response using an ex-vivo interferon-gamma (IFNγ) ELISpot assay with nine HCV genotype-4a overlapping 15-mer peptide pools corresponding to all of the HCV proteins. All subjects were tested for IL28B.rs12979860 SNP by real-time PCR. An HCV-specific CMI was demonstrated in ~27% of the seronegative aviremic HCWs (group 1), suggesting clearance of infection after low-level exposure to HCV. The frequency of IL28B.rs12979860 C allele homozygosity in the four groups was 49%, 48%, 49%, and 23%, while that of the T allele was 14%, 16%, 12 and 19%, respectively, suggesting differential distributions among subjects with different HCV status. As reported, IL28B.rs12979860 predicted the outcome of HCV infection (p < 0.05), but we did not find any relationship between the IL28B genotypes and the outcome of HCV-specific CMI responses in the four groups (p > 0.05). The data show differential IL28B.rs12979860 genotype distribution among Egyptian HCWs with different HCV status and could not predict the outcome of HCV-specific CMI responses.

Interleukin 28B.rs12979860 genotype does not affect hepatitis C viral load in Egyptians with genotype 4 chronic infection.

Several host and viral factors affect the natural history of Hepatitis C Virus (HCV) infection. Interleukin 28B (IL28B).rs12979860 single nucleotide polymorphism (SNP) was found to predict viral clearance with and without therapy. Subjects with the CC (favorable) genotype of IL28B.rs12979860 were more likely to spontaneously clear the infection and respond favorably to therapy. These data suggest that subjects with the "favorable" CC genotype might have a lower viral load when compared to those with the "unfavorable" TT genotype. Therefore, we examined the effect of IL28B.rs12979860 SNP on HCV viral load and clearance among HCV-infected Egyptians. This cross sectional study was conducted on 375 HCV antibody-positive subjects. Detection and quantification of HCV-RNA was determined by RT-PCR. IL28B.rs12979860 genotyping was performed using SYBR green real-time PCR and specific primers. Of 375 HCV-antibody positive subjects, 239 (63.7%) had chronic HCV infection while the remaining 136 (36.3%) subjects had spontaneously cleared the virus. The frequency of IL28-B CC, CT, and TT genotypes among spontaneous resolvers were 54.4%, 39.0%, and 6.6% while among the chronically infected subjects, they were 31.4%, 49.8%, and 18.8%, respectively. As expected, IL28 genotype predicted spontaneous HCV clearance (p < 0.001). The average HCV viral loads were 1.5 ± 0.69 x 10(6), 0.62 ± 0.11 x 10(6) and 0.51 ± 0.14 x 10(6) IU/ml among chronic subjects with the IL28B.rs12979860 CC, CT and TT genotypes, respectively (p > 0.05). In conclusion, our results show that IL28B.rs12979860 genotype does not affect viral load among chronic HCV infected Egyptians. These findings further confirm the complexity of viral host interactions in determining HCV infection outcome.

Design, Synthesis, and Molecular Docking of Novel Pyrrolooxazepinediol Derivatives with Anti-Influenza Neuraminidase Activity.

A series of novel pyrrolo[2,1-b][1,3]oxazepine-8,9-diol derivatives 12-15 were synthesized starting from l-tartaric acid, which was transformed into anhydride which then reacted with allylamine in xylene to afford the imide 2. The target molecules 12-15 were achieved via ring-closing metathesis with the Grubbs catalyst, followed by reduction of the carbonyl group and deprotection of hydroxyl groups. Finally, catalytic hydrogenation of the double bond afforded the title compounds 12-15. Molecular docking study of the title compounds 12-15 was carried out against neuraminidase as the target enzyme, in an attempt to understand the mechanism of action of the tested compounds as potential neuraminidase inhibitors. Molecular docking of the target compounds showed that all tested compounds bind to the active site of neuraminidase, with moderate to high binding energy. Compounds 12-15 were examined for their antiviral activity against H5N1 virus (A/chicken/Egypt/1/2008). Oseltamivir phosphate was used as a control for antiviral activity. The results show that compound 12 (EC50 = 0.016 µg/mL) exhibited potent anti-influenza (H5N1) activity, which approximately equals that of oseltamivir (EC50 = 0.012 µg/mL). Also, it had a therapeutic index similar to that of oseltamivir phosphate (∼20). The data also revealed that compounds 13, 14, and 15 had slightly lower antiviral activity and lower cytotoxicity than oseltamivir phosphate, with LD50 of 0.188, 0.162, and 0.176 µg/mL, respectively. However, 13, 14, and 15 had lower therapeutic indices than 12. In conclusion, we were able to synthesize cheap and potent anti-H5N1 compounds.

Stigma and inequality in the management of coronavirus disease-2019 (COVID-19)- patients among healthcare providers in Saudi Arabia and associated beliefs: A cross sectional study.

BACKGROUND: Stigma and discrimination (SAD) can greatly impact the management of Coronavirus Disease-19 (COVID-19) patients. Studies examining SAD in managing COVID-19 patients in Saudi Arabia (SA) are limited. This study examined the discrimination and inequality in treating COVID-19 patients and the associated beliefs among healthcare workers (HCWs) during the pandemic. METHODS: This was a cross sectional study using an online self-administrated questionnaire by 414 HCWs from September 2022 to January 2023. RESULTS: Of the 414 participants, 53.4% aged 18-30 years, were males (53.4%), mostly Saudi (82.1%), and residents of western SA (90.8%). Almost one-third of the participants (32.1%) were physicians, and 23.7% were pharmacists. 53.1% of the HCWs had work experience < 5 years, 49.3% either had a current/previous COVID-19 infection and 10.1% (n = 42) reported treating ≥ 2000 COVID-19 patients. Most of the participants had not received training on avoiding discrimination against COVID-19 patients (n = 246; 59.4%) and denied receiving any incentives from their institution (n = 294; 71%) or community (n = 248; 59.9%) during the pandemic. Less than 20% of the HCWs had observed some sort of discrimination in providing care for COVID-19 patients. The responses significantly differed by nationality, educational level, specialty, workplace and COVID-19 infection status (P < 0.05). Approximately, 20% of the participants held negative beliefs about COVID-19 patients and these responses varied significantly by nationality, work experience, workplace, gender, and age (p < 0.05). Cross-tabulation and regression analyses showed some differences by gender, workplace, specialty and COVID-19 infection status. CONCLUSIONS: There was a low prevalence of negative beliefs about COVID-19 patients and a low prevalence of discriminatory behaviors in providing care for these patients in SA. Training and governmental regulations could further lower the prevalence of SAD against COVID-19 patients.