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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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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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A Study on Heat Transfer Enhancement through Various Nanofluids in a Square Cavity with Localized Heating

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Energy Engineering 2021年第6期118卷 1659-1679页

作者： Sheikh Hassan Didarul Ahasan redwan md.mamun molla Sharaban Thohura m.Abu Taher Sadia Siddiqa Department of mathematics and Physics North South UniversityDhakaBangladesh Center for Applied and Scientific Computing(CASC) North South UniversityDhakaBangladesh Department of mathematics Jagannath UniversityDhakaBangladesh Department of mathematics Dhaka University of Engineering and TechnologyGazipurBangladesh Department of mathematics COMSAT University IslamabadAttock CampusAttockPakistan

A two-dimensional(2D)laminar flow of nanofluids confined within a square cavity having localized heat source at the bottom wall has been *** governing Navier–Stokes and energy equations have been non dimensionalized using the appropriate non dimensional variables and then numerically solved using finite volume *** flow was controlled by a range of parameters such as Rayleigh number,length of heat source and nanoparticle volume *** numerical results are represented in terms of isotherms,streamlines,velocity and temperature distribution as well as the local and average rate of heat transfer.A comparative study has been conducted for two different base fluids,ethylene glycol and water as well as for two different solids Cu and Al_(2)O_(3).It is found that the ethylene glycol-based nanofluid is superior to the water-based nanofluid for heat transfer enhancement.

关键词： Nanofluids finite volume method heat transfer enhancement localized heating

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Oral Health Status and Gingival Response to Three Different Restorative materials among Saudi Patients: A Clinical &Histopathological Study

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International Journal of Clinical medicine 2019年第2期10卷 78-90页

作者： mohammed m. A. Abdullah Al-Abdaly Hassan mohammed Al-Harthi Salem mohammed Al-Harthi redwan Abdullah Ali Almalki Periodontics and Community Dental Sciences Department College of Dentistry King Khalid University Abha Saudi Arabia College of Dentistry King Khalid University Abha Saudi Arabia

Background: The correlation between oral health and dental restoration is fundamental. For the gingival and periodontal tissues to stay healthy, dental restoration should be in regularity with the surrounding tissues. This study aims to assess the oral health status and histopathological gingival response to three different restorative materials among Saudi patients. methods: The study groups consist of 240 patients (50% males and 50% females), aged 18 - 45, with inclusion and exclusion criteria in the study. Participants are divided into three equal groups: those with composite resin restorations, those with amalgam restorations and those with glass ionomer restorations. Biopsies were taken from adjacent gingival tissues. Clinical parameters were determined by: plaque index (PLI), gingival index (GI) and clinical attachment loss (CAL). All data were collected and evaluated by through statistical analysis. Results: The clinical findings of the current study revealed that amalgam restorations produce a higher means of PLI, GI and CAL compared with composite resin restorations and glass ionomer restorations, but not insignificant levels, except CAL (p = 0.004*). As for histopathological findings, there were significant differences in gingival tissue response to amalgam restorations, composite resin restorations and glass ionomer cement fillings, where there were statistically significant differences in numbers of chronic inflammatory cells (p Conclusion: At the end of the present study, we concluded that the amalgam restorations are less biocompatible compared to composite resin restorations and glass ionomer restorations.

关键词： Gingival Response Histopathological Study Oral Health Status

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