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Flow cytometric detection of hepatitis C virus antigens in infected peripheral blood leukocytes: Binding and entry

World Journal of Gastroenterology 2005年第33期11卷 5203-5208页

作者： Mostafa K El-Awady Ashraf A Tabll El-Rashdy M Redwan Samar Youssef moataza h omran Fouad Thakeb Maha El-Demellawy Department of Biomedical Technology National Research Center Tahrir Street Dokki Cairo Egypt Genetic Engineering and Biotechnology Research Institute New Borg El-Arab City Alexandria Egypt Department of Gastroenterology-hepatology Faculty of Medicine Cairo University Egypt Genetic Engineering and Biotechnology Research Institute New Borg El-Arab City Alexandria Egypt

AIM： We designed two synthetic-core-specific peptides core 1 （C1） and core 2 （C2）, and an E1-specific peptide （El）. We produced specific polyclonal antibodies against these peptides and used the antibodies for detection of hCV antigens on surface and within infected peripheral blood leukocytes. METhODS： Peripheral blood from a healthy individual who tested negative for hCV RNA was incubated with hCV type 4 infected serum for i h and 24 h at 37 ℃. Cells were stained by direct and indirect immunofluorescence and measured by flow cytometry. RESULTS： After 1 h of incubation, antibodies against C1, C2, and El detected hCV antigens on the surface of 27%, 26% and 73% of monocytes respectively, while 10%, 5% and 9% of lymphocytes were positive with anti-C1, anti-C2 and anti-E1 respectively. Only 1-3% of granulocytes showed positive staining with anti-C1, anti-C2 and anti E1 antibodies. After 24 h of incubation, we found no surface staining with anti-C1, anti-C2 or anti-E1. Direct immunostaining using anti-C2 could not detect intracellular hCV antigens, after 1 h of incubation with the virus, while after 24 h of incubation, 28% of infected cells showed positive staining. Only plus strand RNA was detectable intracellularly as early as 1 h after incubation, and remained detectable throughout 48 h post-infection. Interestingly, minus RNA strand could not be detected after 1 h, but became strongly detectable intracellularly after 24 h post-infection. CONCLUSION： Monocytes and lymphocytes are the preferred target cells for hCV infection in peripheral blood leukocytes. Our specific anti-core and anti-E1 antibodies are valuable reagents for demonstration of hCV cell cycle. Also, hCV is capable of infecting and replicating in peripheral blood mononuclear cells as confirmed by detection of minus strand hCV RNA as well as intracellular staining of core hCV antigen.

关键词： Flow cytometry hepatitis C virus Envelope Core Antibodies Indirect immunofluorescence Minus and plus RNA strand Peripheral blood mononudear cells

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Positional effect of mutations in 5'UTR of hepatitis C virus 4a on patients' response to therapy

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World Journal of Gastroenterology 2009年第12期15卷 1480-1486页

作者： Mostafa K El Awady hassan M Azzazy Ahmed M Fahmy Sherif M Shawky Noha G Badreldin Samar S Yossef moataza h omran Abdel Rahman N Zekri Said A Goueli Department of Biomedical Technology National Research CenterDokkiCairo 12622Egypt Department of Chemistry and Yousef Jameel Science and Technology Research Center the American University in CairoCairo 11511Egypt Structural biology department Free University of AmsterdamDe Boelelaan 10851081 HV AmsterdamThe Netherlands National Cancer Institute Cairo 11796Egypt Department of Pathology and Laboratory Medicine University of WisconsinMadisonWI 53711United States

AIM： To investigate the effects of mutations in domain Ⅲ of the hepatitis C virus （hCV） internal ribosome entry sequences （IRES） on the response of chronic hCV genotype 4a patients to interferon therapy.METhODS： hCV RNA was extracted from 19 chronic hCV 4a patients receiving interferon/ribavirin therapy who showed dramatic differences in their response to combination therapy after initial viral clearance. IRES domain Ⅲ was cloned and 15 clones for each patient were sequenced. The obtained sequences were aligned with genotype 4a prototype using the ClustaIW program and mutations scored. Prediction of stem-loop secondary structure and thermodynamic stability of the major quasispecies in each patient was performed using the MFOLD 3.2 program with Turner energies and selected constraints on base pairing.RESULTS： Analysis of RNA secondary structure revealed that insertions in domain Ⅲ altered WatsonCrick base pairing of stems and reduced molecular stability of RNA, which may ultimately reduce binding affinity to ribosomal proteins. Insertion mutations in domain - were statistically more prevalent in sustained viral response patients （SVR, n = 14） as compared to breakthrough （BT, n = 5） patients.CONCLUSION： The influence of mutations within domain Ⅲ on the response of hCV patients to combination therapy depends primarily on the position, but not the frequency, of these mutations within IRES domain Ⅲ.

关键词： hepatitis C virus Internal ribosome entrysequences Domain Ⅲ Genotype 4a Ribosomal sub-unit Interferon therapy RNA folding

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Antibody to El peptide of hepatitis C virus genotype 4 inhibits virus binding and entry to hepG2 cells in vitro

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World Journal of Gastroenterology 2006年第16期12卷 2530-2535页

作者： Mostafa K EL-Awady Ashraf A Tabll Khaled Atef Samar S Yousef moataza h omran Yasmin EI-Abd Noha G Bader-Eldin Ahmad M Salem Samir F Zohny Wael T EI-Garf Department of Biomedical Technology National Research CenterCairoEgypt Department of Biochemistry Faculty of ScienceAin Shams UniversityCairoEgypt

AIM： To analyze the neutralizing activity of antibodies against E1 region of hepatitis C virus （hCV）. Specific polydonal antibody was raised via immunization of New Zealand rabbits with a synthetic peptide that had been derived from the E1 region of hCV and was shown to be highly conserved among hCV published genotypes. METhODS： hyper-immune hCV E1 antibodies were incubated over night at 4 ℃ with serum samples positive for hCV RNA, with viral loads ranging from 615 to 3.2 million IU/mL. Treated sera were incubated with hepG2 cells for 90 min. Blocking of viral binding and entry into cells by anti E1 antibody were tested by means of RTPCR and flow cytometry. RESULTS： Direct immunostaining using FITC conjugated E1 antibody followed by Flow cytometric analysis showed reduced mean fluorescence intensity in samples pre-incubated with E1 antibody compared with untreated samples. Furthermore, 13 out of 18 positive sera （72%） showed complete inhibition of infectivity as detected by RT-PCR. CONCLUSION： In house produced E1 antibody, blocks binding and entry of hCV virion infection to target cells suggesting the involvement of this epitope in virus binding and entry. Isolation of these antibodies that block virus attachment to human cells are useful as therapeutic reagents.

关键词： Flow cytometry hepatitis C virus E1 envelope Therapeutic antibodies Direct immuno-fluorescence hepG2 cells

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Association between low molecular polypeptide 7 single nucleotide polymorphism and response to therapy in hepatitis C virus infection

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World Journal of hepatology 2013年第3期5卷 97-103页

作者： moataza h omran Basma E Fotouh Samar S Youssef Noha E Ibrahim Wael Nabil EL-Sayed M Mahdy Wafaa G Shosha Mostafa K El-Awady Microbial Biotechnology Department Genetic Engineering Division National Research Center Chemistry Department Faculty of Science Helwan University

AIM: To investigate the relationship between low molecular polypeptide-7 (LMP-7) gene polymorphism and response to interferon (IFN) therapy in chronic hepatitis C virus (hCV) patients. METhODS: LMP-7 polymorphism at codon 49 with nucleotide substitution from A to C was amplified in 104 chronic hCV patients of genotype 4. The amplicons were digested with restriction endonuclease Bsm I and the produced restriction fragment length polymorphism was analyzed. Patients received IFN + regional blood volume therapy for 48 wk and the frequency of thissingle nucleotide polymorphism (SNP) was statistically correlated with treatment response. The exclusion criteria for these patients were stated by the national health program for treating viral hepatitis. Main exclusion criteria included co-infection with hepatitis B virus or schistosomiasis, thyroid dysfunction, uncontrolled diabetes mellitus, history of long term drug or alcohol intake and autoimmune hepatitis. Multivariate analyses were done to correlate LMP-7 SNP plus several factors such as age, gender, weight, serum alpha-fetoprotein (AFP) and alanine aminotransferase levels, liver activity, fibrosis score and viral load with response to therapy. RESULTS: The data presented in this study clearly demonstrated statistically significant differences between sustained virological response (SVR) (defined as the absence of hCV RNA levels in the patient's sera at least 6 mo after discontinuation of treatment) and non-response (NR) (where hCV RNA levels in the patient's sera never become undetectable for 6 mo during or after treatment). Variables were described as odds ratio with 95%CI. The data were considered significant if P values were ≤ 0.05; highly significant if P highly significant if P howed that 91.7% of patients carrying LMP-7 C/C allele were associated with SVR, while the other two genotypes C/A and A/A were associated with NR patients, 83.3% and 64.3% respectively, showing that g

关键词： hepatitis C virus Interferon therapy Low molecular mass polypeptide host gene Single nucleotide polymorphism

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hepG2 cells support viral replication and gene expression of hepatitis C virus genotype 4 in vitro

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World Journal of Gastroenterology 2006年第30期12卷 4836-4842页

作者： Mostafa K El-Awady Ashraf A Tabll Yasmine S El-Abd Mahmoud M Bahgat hussein A Shoeb Samar S Youssef Noha G Bader El Din El-Rashdy M Redwan Maha El-Demellawy moataza h omran Wael T El-Garf Said A Goueli Department of Biomedical Technology National Research Center Tahrir Street PO 12622 Dokki Cairo Egypt Department of Medicinal Chemistry Na- tional Research Center Tahrir Street PO 12622 Dokki Cairo Egypt Department of Microbiology and Immunology Faculty of Pharmacy Cairo University Cairo Egypt Genetic Engineering and Biotechnology Research Institute New Borg El-Arab City Alexandria Egypt Department of Pathology School of Medicine University of Wisconsin Madison WI United States

AIM： TO establish a cell culture system with longterm replication of hepatitis C virus （hCV） genome and expression of viral antigens in vitro. METhODS： hepG2 cell line was tested for its susceptibility to hCV by incubation with a serum from a patient with chronic hepatitis C. Cells and supernatant were harvested at various time points during the culture. Culture supernatant was tested for its ability to infect na＇ive cells. The presence of minus （antisense） RNA strand, and the detection of core and E1 antigens in cells were examined by RT-PCR and immunological techniques （flow cytometry and Western blot） respectively. RESULTS： The intracellular hCV RNA was first detected on d 3 after infection and then could be consistently detected in both cells and supernatant over a period of at least three months. The fresh cells could be infected with supernatant from cultured infected cells. Flow cytometric analysis showed surface and intracellular hCV antigen expression using in house made polyclonal antibodies （anti-core, and anti-E1）. Western blot analysis showed the expression of a cluster of immunogenic peptides at molecular weights extended between 31 and 45 kDa in an one month old culture of infected cells whereas this cluster was undetectable in uninfected hepG2 cells. CONCLUSION： hepG2 cell line is not only susceptible to hCV infection but also supports its replication in vitro. Expression of hCV structural proteins can be detected in infected hepG2 cells. These cells are also capable of shedding viral particles into culture media which in turn become infectious to uninfected cells.

关键词： hepatitis C virus In vitro propagation Genomic replication Gene expression hepG2 cells

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Noninvasive assessment of hepatic fibrosis in Egyptian patients with chronic hepatitis C virus infection

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World Journal of Gastroenterology 2012年第23期18卷 2988-2994页

作者： Shawky Abdelhamid Fouad Serag Esmat Dalia omran Laila Rashid Mohamed h Kobaisi Department of Internal Medicine Faculty of Medicine Cairo University Cairo 11562 Egypt Department of Tropical Medicine Faculty of Medicine Cairo University Cairo 11562 Egypt Department of Biochemistry Faculty of Medicine Cairo University Cairo 11562 Egypt Department of Pathology National Institute of Nephrology Cairo 35789 Egypt

AIM: To evaluate the accuracy of specific biochemical markers for the assessment of hepatic fibrosis in patients with chronic hepatitis C virus (hCV) infection. METhODS: One hundred and fifty-four patients with chronic hCV infection were included in this study; 124 patients were non-cirrhotic, and 30 were cirrhotic. The following measurements were obtained in all patients: serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin, total bilirubin, prothrombin time and concentration, complete blood count, hepatitis B surface antigen (hBsAg), hCVAb, hCV-RNA by quantitative polymerase chain reaction, abdominal ultrasound and ultrasonic-guided liver biopsy. The following ratios, scores and indices were calculated and compared with the results of the histopathological examination: AST/ALT ratio (AAR), age platelet index (API), AST to platelet ratio index (APRI), cirrhosis discriminating score (CDS), Pohl score, G teborg University Cirrhosis Index (GUCI). RESULTS: AAR, APRI, API and GUCI demonstrated good diagnostic accuracy of liver cirrhosis (80.5%, 79.2%, 76.6% and 80.5%, respectively); P values were: he studied parameters, AAR and GUCI gave the highest diagnostic accuracy (80.5%) with cutoff values of 1.2 and 1.5, respectively. APRI, API and GUCI were significantly correlated with the stage of fibrosis (P he grade of activity (P hile CDS only correlated significantly with the stage of fibrosis (P h the degree of activity (P > 0.05). In addition, we found significant correlations for the AAR, APRI, API, GUCI and Pohl score between the non-cirrhotic (F0, F1, F2, F3) and cirrhotic (F4) groups (P values: 0.05). CONCLUSION: The use of AAR, APRI, API, GUCI and Pohl score measurements may decrease the need

关键词： Age platelet index Aspartate aminotrans-ferase platelet ratio index Aspartate aminotransferase-to-alanine aminotransferase ratio Cirrhosis discrimi-nating score Fibrosis evaluation G^teborg UniversityCirrhosis Index hepatitis C virus infection Liver fibro-sis Pohl score

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Structural, electronic, elastic, and thermal properties of CaNih3 perovskite obtained from first-principles calculations

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Chinese Physics B 2018年第3期27卷 384-390页

作者： S Benlamari h Bendjeddou R Boulechfar SAmara Korba h Meradji R Ahmed S Ghemid R Khenata S Bin omran Laboratoire LPR Departement de Physique Faculté des Sciences Université Badji Mokhtar Annaba Algeria Department of Physics Faculty of Science Universiti Teknologi Malaysia UTM Skudai 81310 Johor Malaysia Laboratoire de Physique Quantique et de Modelisation Mathematique de la Matiere (LPQ3M) Universite de Mascara-29000-Algeria Department of Physics and Astronomy College of Science King Sand University P.O. Box 2455 Riyadh 11451 Saudi Arabia

A theoretical study of the structural, elastic, electronic, mechanical, and thermal properties of the perovskite-type hydride CaNih3 is presented. This study is carded out via first-principles full potential （FP） linearized augmented plane wave plus local orbital （LAPW＋lo） method designed within the density functional theory （DFT）. To treat the exchange-correlation energy/potential for the total energy calculations, the local density approximation （LDA） of Perdew-Wang （PW） and the generalized gradient approximation （GGA） of Perdew-Burke-Ernzerhof （PBE） are used. The three independent elastic constants （C11, C12, and C44） are calculated from the direct computation of the stresses generated by small strains. Besides, we report the variation of the elastic constants as a function of pressure as well. From the calculated elastic constants, the mechanical character of CaNih3 is predicted. Pertaining to the thermal properties, the Debye temperature is estimated from the average sound velocity. To further comprehend this compound, the quasi-harmonic Debye model is used to analyze the thermal properties. From the calculations, we find that the obtained results of the lattice constant （ao）, bulk modulus （Bo）, and its pressure derivative （B0′） are in good agreement with the available theoretical as well as experimental results. Similarly, the obtained electronic band structure demonstrates the metallic character of this perovskite-type hydride.

关键词： perovskite-type hydrides elastic constants hydrogen storage materials

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An advanced workflow to compress the uncertainties of stochastic distribution of Bahariya reservoir properties using 3D static modeling:An example from heba Oil Fields,Western Desert,Egypt

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Petroleum Research 2023年第2期8卷 205-216页

作者： Tamer hassan Ahmad M.K.Basal Mohammad A.omran Manar h.Mowafy Mohammad A.Sarhan Qarun Petroleum Company(Apache J.V) New MaadiCairo11742Egypt Geology Department Faculty of ScienceDamietta University34517Egypt

The main objective of this paper is to construct a static model that compress the uncertainties of the stochastic distribution of the reservoir properties of the Bahariya Formation in heba field,at the northeastern portion of the Western *** model has been constructed through the integration of the interpretations of the eighteen 2D seismic sections and the analysis of well logs data for four wells(hEBA 300X,***-E-1X,***-D-1X,and hEBA 10X)drilled in the study *** set of data was implemented in a harmonic *** framework was the first step created on the basis of the seismic and well log *** zonation was mainly managed by the marine flooding events took place during the Cenomanian *** trapping faults position uncertainty has been compressed through the tying of the seismic profiles with the identified fault cuts in the well *** porosity spectrum was broke up into three reservoir *** results showed heterogeneous facies qualities for oil production in specific five zones in the topmost part of the Bahariya *** effective porosity model was generated stochastically considering the normal distribution for each reservoir *** saturation was distributed by two methods;1)Sequential Gaussian Simulation that was co-simulated by porosity model.2)Log-based saturation height function for each reservoir *** methodology provided as accurate as possible estimates for the volume calculation by quantifying the sensitivity of the important parameters such as oil ***,the model was prepared to be used as a front end for dynamic simulation.

关键词： 3D static modeling Stochastic distribution Bahariya formation Reservoir characterization heba oil fields

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Predictors of anti-SARS-CoV-2 seropositivity:An Egyptian population-based study

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Infectious Medicine 2022年第2期1卷 113-123页

作者： Engy Mohamed El-Ghitany Ayat Ashour Azza Galal Farghaly Mona h.hashish Eman A.omran Department of Tropical health High Institute of Public HealthAlexandria UniversityAlexandriaEgypt Department of Family health High Institute of Public HealthAlexandria UniversityAlexandriaEgypt Department of Microbiology High Institute of Public HealthAlexandria UniversityAlexandriaEgypt

Background:Population-based studies on the determinants of COVID-19 seroprevalence constitute a cornerstone in guiding appropriate preventive *** studies are scarce in Egypt,thus we conducted this study to explore risk factors for SARS-CoV-2 ***:This survey included 2919 participants from 10 Egyptian *** were tested for SARS-CoV-2 spike(S)and nucleocapsid(N)*** and multivariate analyses were performed to identify associated factors and predictors of seropositivity regarding sociodemographic factors,clinical data,and personal practices of participants.A subgroup analysis was performed to investigate the occupational risks of ***:Seropositivity was recorded in 1564 participants(53.6%).Independent predictors of seropositivity in-cluded non-smokers(aOR=1.817;95%CI:1.407-2.346,p=0.000),having blood group A(aOR=1.231;95%CI:1.016-1.493,p=0.034),a history of COVID-19 infection(aOR=2.997;95%CI:2.176-4.127,p=0.000),COVID-19 vaccination(aOR=4.349;95%CI:2.798-6.759,p=0.000),higher crowding index(aOR=1.229;95%CI:1.041-1.451,p=0.015),anosmia and/or ageusia(aOR=3.453;95%CI:2.661-4.481,p=0.000)and history of fever(aOR=1.269;95%CI:1.033-1.560,p=0.023).healthcare worker and Obesity/overweight were additional significant predictors of seropositivity among the working participants(aOR=1.760;95%CI:1.301-2.381,p=0.000 and aOR=1.384;95%CI:1.059-1.808,p=0.019,respectively).Additional factors showing association with seropositivity in the univariate analysis were:female gender,age group(15-39 years),higher educational level(preparatory and above),lack of environmental disinfection and having roommates at the *** was a positive correlation between the titers of both *** was weakly corre-lated with anti-S titer,while anti-N was significantly correlated with the number of protective measures applied by the *** antibodies were significantly correlated with adult BMI,while both we

关键词： COVID-19 Predictors Seroprevalence Anti-spike Anti-nucleocapsid

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Preparation of Dried Vegetarian Soup Supplemented with Some Legumes

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Food and Nutrition Sciences 2014年第22期5卷 2274-2285页

作者： Amal M. h. Abdel-haleem Azza A. omran Department of Crops Technology Research Food Technology Research Institute Agricultural Research Center Giza Egypt

The present work was aimed to prepare dried vegetarian soup supplemented with some legumes. Potatoes, hull-less barley flour, carrot, tomatoes, onion, garlic, salt, black pepper, coriander and cumin (served as F1) were used in combination with lentil, green pea and chickpea to formulate F2, F3, and F4, respectively. Chemical, physical, rheological and sensory evaluation was performed. The results indicated that supplementation with legumes significantly enhanced the nutritional characteristics, where the dried vegetarian soup mixtures had reasonable amounts of the required nutrients particularly, protein, carbohydrates, fats, Fe and Zn with good in vitro protein digestibility and mineral availability. Moisture content and water activity predicted the extended shelf-life and stability of the dried soup mixtures. The four resultant soup samples F1, F2, F3 and F4 had a noticeable viscosity pattern characterized by a non-Newtonian pseudoplastic flow behavior. Supplementation with legumes significantly (p ≤ 0.05) affects taste, color, flavor attributes and overall acceptability of the resultant soup samples. But, it had no significant effect on thickness and appearance. The results clearly demonstrated the usefulness of supplementing the dried vegetarian soup mixtures with legumes to enhance nutritional and technological quality of the resultant soup and lentil was the most valuable addition with the highest acceptability.

关键词： Vegetarians Soup Legumes Nutritional Quality Physical Characteristics Rheological Properties Sensory Quality Attributes

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