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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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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 < 0.01 and very highly significant if P < 0.001. Current data showed 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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Assessment of circulating levels of microRNA-326,microRNA-424,and microRNA-511 as biomarkers for hepatocellular carcinoma in Egyptians

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World Journal of Hepatology 2022年第8期14卷 1562-1575页

作者： samar Samir youssef Asmaa Elfiky Mohamed M Nabeel Hend Ibrahim Shousha Tamer Elbaz Dalia Omran Mohammad Saeed Marie Mohammad A Elzahry Amr Abul-Fotouh Ahmed Hashem Mohamed F Guda Ashraf O Abdelaziz Department of Microbial Biotechnology National Research CentreCairo 1211Egypt Department of Environmental and Occupational Medicine National Research CentreCairo 1211Egypt Department of Endemic Medicine and Hepatogastroenterology Faculty of MedicineCairo UniversityCairo 11562 Egypt Department of Endemic Medicine Faculty of MedicineCairo UniversityCairo 1256Egypt Theodor Bilharz Research Institute Cairo 1256Egypt

BACKGROUND Hepatocellular carcinoma(HCC)is the fifth most common *** expression of microRNAs(miRNAs)-326,miRNA-424,and miRNA-511 has been associated with the diagnosis and prognosis of HCC in different ***,limited information is available regarding their expression in Egyptian HCC *** To assess the role of circulating miRNAs-326,miRNA-424,and miRNA-511 in Egyptian HCC *** This prospective observational study included 70 HCC patients and 25 healthy *** circulating levels of these three miRNAs were evaluated by real-time *** operating characteristic curve analysis was used to test the diagnostic accuracy of micro RNA expression *** All miRNAs were differentially expressed in HCC patients;miRNAs326 and miRNA-424 were upregulated,while miRNA-511 was *** miRNA-326 and miRNA-424 showed sensitivity and specificity of 97%,71.4%,and 52%,60%,respectively,to differentiate HCC from ***,miRNA-326 was associated with survival and could differentiate between Child grades(A vs B);miRNA-424 significantly differentiated early vs intermediate stages of HCC;while miRNA-511 was significantly correlated with response to modified Response Evaluation Criteria in Solid Tumors(m RECIST).CONCLUSION We conclude that miRNA-326,miRNA-424,and miRNA-511 have diagnostic and prognostic roles in Egyptian patients with hepatitis C virus-related HCC and should be considered for better disease management.

关键词： Hepatocellular carcinoma miRNAs-326 miRNA-424 miRNA-511 Modified response evaluation criteria in solid tumors

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Phosphoric Acid Pretreatment and Saccharification of Paper Sludge as a Renewable Material for Cellulosic Fibers

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Journal of Renewable Materials 2024年第9期12卷 1573-1591页

作者： samar El-Mekkawi Wafaa Abou-Elseoud Shaimaa Fadel Enas Hassan Mohammad Hassan Chemical Engineering and Pilot Plant Department National Research CentreDokkiGiza12622Egypt Cellulose and Paper Department National Research CentreDokkiGiza12622Egypt

Recycling of paper sludge waste is crucial for establishing a sustainable green industry. This waste contains valuable sugars that can be converted into important chemicals such as ethanol, poly hydroxybutyrate, and lactic acid. However, the main challenge in obtaining sugars in high yield from paper sludge is the high crystallinity of cellulose, which hinders hydrolysis. To address this, pretreatment using phosphoric acid was optimized using response surface methodology to facilitate cellulose hydrolysis with minimal energy and chemicals. The created prediction model using the response surface method considered factors such as acid concentration (ranging from 60% to 85%), consistency (ranging from 4% to 10%), temperature (ranging from 25°C to 80°C), and time (ranging from 0.5 to 4 h). The results revealed that the model’s significant factors affecting the yield were acid concentration, reaction time, temperature, and the product of acid concentration and temperature, while the model’s significant factors affecting the crystallinity were the consistency, the temperature, and their product. The results showed that the optimum conditions for pretreatment were using an acid concentration of 64%, temperature of 25°C, consistency of 10%, and time of 30 min. The hydrolysis of the conditionally pretreated paper sludge resulted in a weight loss of 42%, compared to only 18% weight loss in non-pretreated paper sludge. Furthermore, the optimized conditions led to low levels of furfurals and acetic acid, which are undesirable by-products that can interfere with sugar fermentation. The total sugar obtained under the optimized conditions was 0.43 g glucose/g sample (10.46 g/L), while the contents of methyl furfural, furfural, and acetic acid were 21.65, 235.7, and 4.57 mg/L, respectively. This study demonstrates the potential of phosphoric acid for pretreatment and hydrolysis of paper sludge, enabling efficient saccharification and the production of sugars with minimal undesired

关键词： Waste valorization fermentable sugars response surface methodology acid hydrolysis cellulose crystallinity

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Unique presentation of neonatal liver failure:A case report

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World Journal of Clinical Pediatrics 2024年第2期13卷 221-226页

作者： Eman Al Atrash Amer Azaz samar Said Mohammad Miqdady Department of Pediatrics Division of Pediatric GastroenterologySheikh Khalifa Medical CityAbu Dhabi 971United Arab Emirates Department of Pathology Mayo ClinicRochesterMN 55905United States

BACKGROUND Acute fulminant liver failure rarely occurs in the neonatal *** etiologies include viral infection(15%),metabolic/genetic disease(10%),hematologic disorders(15%),and ischemic injury(5%).Gestational alloimmune liver disease usually manifests as severe neonatal liver failure,with extensive hepatic and extrahepatic iron overload,sparing the reticuloendothelial *** liver failure is a rare cause of liver failure where a patient presents with liver failure in the neonatal period with no hepatocytes in liver *** SUMMARY A 5-week-old male presented with *** examination revealed an alert but deeply icteric *** data demonstrated direct hyperbilirubinemia,a severely deranged coagulation profile,normal transaminase,and normal *** resonance imaging of the abdomen was suggestive of perinatal *** biopsy showed histiocytic infiltration with an absence of *** hemosiderin deposition was identified in a buccal mucosa *** Neonatal liver failure in the absence of hepatocellular regeneration potentially reflects an acquired or inborn defect in the regulation of hepatic regeneration.

关键词： Liver Hyperbilirubinemia Le foie vide Neonatal hemochromatosis Case report

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Contrast Normalization Strategies in Brain Tumor Imaging:From Preprocessing to Classification

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Computer Modeling in Engineering & Sciences 2024年第8期140卷 1539-1562页

作者： samar M.Alqhtani Toufique A.Soomro Faisal Bin Ubaid Ahmed Ali Muhammad Irfan Abdullah A.Asiri Department of Information Systems College of Computer Science and Information SystemsNajran UniversityNajran61441Saudi Arabia Department of Electronic Engineering The University of LarkanoLarkanaSindh75660Pakistan Computer Science Department Sukkur IBA UniversitySukkurSindh65200Pakistan Eletrical Engineering Department Sukkur IBA UniversitySukkurSindh65200Pakistan Electrical Engineering Department College of EngineeringNajran UniversityNajran61441Saudi Arabia Radiological Sciences Department College of Applied Medical SciencesNajran UniversityNajran61441Saudi Arabia

Cancer-related to the nervous system and brain tumors is a leading cause of mortality in various *** resonance imaging(MRI)and computed tomography(CT)are utilized to capture brain *** plays a crucial role in the diagnosis of brain tumors and the examination of other brain ***,manual assessment of MRI images by radiologists or experts is performed to identify brain tumors and abnormalities in the early stages for timely ***,early diagnosis of brain tumors is intricate,necessitating the use of computerized *** research introduces an innovative approach for the automated segmentation of brain tumors and a framework for classifying different regions of brain *** proposed methods consist of a pipeline with several stages:preprocessing of brain images with noise removal based on Wiener Filtering,enhancing the brain using Principal Component Analysis(PCA)to obtain well-enhanced images,and then segmenting the region of interest using the Fuzzy C-Means(FCM)clustering technique in the third *** final step involves classification using the Support Vector Machine(SVM)*** classifier is applied to various types of brain tumors,such as meningioma and pituitary tumors,utilizing the Contrast-Enhanced Magnetic Resonance Imaging(CE-MRI)*** proposed method demonstrates significantly improved contrast and validates the effectiveness of the classification framework,achieving an average sensitivity of 0.974,specificity of 0.976,accuracy of 0.979,and a Dice Score(DSC)of ***,this method exhibits a shorter processing time of 0.44 s compared to existing *** performance of this method emphasizes its significance when compared to state-of-the-art methods in terms of sensitivity,specificity,accuracy,and *** enhance the method further in the future,it is feasible to standardize the approach by incorporating a set of classifiers to increase the robustness of the brain classi

关键词： Brain tumor magnetic resonance imaging principal component analysis fuzzy c-clustering support vector machine

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Neuroprotective effect of liraglutide and memantine in a rat model of Alzheimer’s disease

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Asian Pacific Journal of Tropical Biomedicine 2024年第5期14卷 207-214页

作者： Duaa M.Bakhshwin Foziya W.Alhalabi Mohamad Omar Barasheed Ahmed M.Bakhshwin Maha H.Jamal Mohammed A.Bazuhair Fatemah O.Kamel Rania M.Magadmi Zoheir A Damanhouri samar M.Alsaggaf Department of Clinical Pharmacology Faculty of MedicineKing Abdulaziz UniversityJeddahSaudi Arabia Hera General Hospital MakkahSaudi Arabia Department of Pathology Faculty of MedicineKing Abdulaziz UniversityJeddahSaudi Arabia Department of Clinical Anatomy Faculty of MedicineKing Abdulaziz UniversityJeddahSaudi Arabia

Objective:To assess the effect of memantine combined with liraglutide on aluminum chloride(AlCl_(3))and D-galactose(D-GAL)-induced neurotoxicity in ***:Male Wistar rats were divided into 5 groups of 5 animals each:the positive control,the negative control,the memantine-treated group,the liraglutide-treated group,and the combination group treated with memantine and ***_(3)and D-GAL were used to induce *** tests,brain beta-amyloid protein,and oxidative stress biomarkers were ***:The Morris water maze test indicated an enhanced memory in the combination ***,the combination treatment of liraglutide and memantine resulted in a remarkable reduction in the beta-amyloid protein level in the brain *** inflammation and oxidative stress biomarkers were significantly reduced,and the levels of antioxidant parameters were ***:The combination of liraglutide and memantine exerts neuroprotective effects and enhances memory and cognitive functions in rats with Alzheimer’s disease.

关键词： Alzheimer’s disease Cognitive function Glucagonlike peptide-1 Liraglutide Memantine

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Case Report and Clinical Management of a Case of Osteogenesis Imperfecta Detected in the Prenatal Period

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Open Journal of Obstetrics and Gynecology 2024年第7期14卷 996-1002页

作者： Amina Chaieb Oumayma Ben Rejeb samar Knaz Yasmine Ben Ali Syrine Chelly Safia Ernez Mouna Derouiche Department of Gynecology and Obstetrics Farhat Hached University Hospital Sousse Tunisia Faculty of Medicine Ibn Al Jazzar University of Sousse Sousse Tunisia

Osteogenesis imperfecta is a hereditary disease characterized by bone fragility due to a defect in type I collagen synthesis. The diagnosis is typically suspected based on suggestive ultrasound findings and confirmed through genetic studies. We present a case of osteogenesis imperfecta suspected during obstetrical ultrasound at 19 weeks’ gestation, which was later confirmed radiographically through computed tomography. Due to the severity of the condition, therapeutic termination of pregnancy was indicated.

关键词： Osteogenesis Imperfecta Ultrasound Screening Antenatal Diagnosis

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Association analysis of FTO gene polymorphisms and obesity risk among Egyptian children and adolescents

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Genes & Diseases 2017年第3期4卷 170-175页

作者： samar Samir Abdelmajed Mohamed youssef Moushira Erfan Zaki Naglaa Abu-Mandil Hassan Somaia Ismail October University for Modern Sciences and Arts(MSA) GizaEgypt Organic chemistry Department Faculty of scienceCairo UniversityEgypt Biological Anthropology Department Medical Research DivisionNational Research CentreEgypt Medical Molecular Genetics Department Human Genetics and Genome Research DivisionNational Research CentreGizaEgypt

Obesity is a common disorder that has a significant impact on human health as it may lead to many serious diseases and sometimes *** genome-wide association studies(GWAS)confirmed that there is a relationship between some variants in the first intron of the fat mass and obesity associated(FTO)gene and obesity in adults and children in different ethnic *** our study,the association of the FTO rs9939609 and rs17817449 variants with obesity was investigated in Egyptian children and *** examined rs9939609 and rs17817449 polymorphisms in 100 control and 100 obese cases,we used the restriction fragment length polymorphism(RFLP)technique to genotype the *** current study showed that there were no significant differences(P>0.05)between the cases and controls in both variants of rs17817449 and rs9939609 ***,there were significant correlations between rs17817449 and cholesterol and between rs9939609 and *** Current Study although the two variants(rs9939609 and rs17817449)didn’t show an association with obesity,but there was a correlation between the lipid profile and these two variants.

关键词： Children Egyptians Fat mass and obesity associated gene(FTO) Obesity Variants

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