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2022
Motawi, T. M. K., N. A. H. Sadik, D. Sabry, S. A. Fahim, and N. N. Shahin, "rs62139665 Polymorphism in the Promoter Region of EpCAM Is Associated With Hepatitis C Virus-Related Hepatocellular Carcinoma Risk in Egyptians", Frontiers in Oncology, vol. 11, pp. 754104, 2022. AbstractWebsite

Hepatocellular carcinoma (HCC) is a universal health problem that is particularly alarming in Egypt. The major risk factor for HCC is hepatitis C virus (HCV) infection which is a main burden in Egypt. The epithelial cell adhesion molecule (EpCAM) is a stem cell marker involved in the tumorigenesis and progression of many malignancies, including HCC. We investigated the association of -935 C/G single nucleotide polymorphism in EpCAM promoter region (rs62139665) with HCC risk, EpCAM expression and overall survival in Egyptians. A total of 266 patients (128 HCV and 138 HCC cases) and 117 age- and sex-matched controls participated in this study. Genotyping, performed using allelic discrimination and confirmed by sequencing, revealed a significant association between EpCAM rs62139665 and HCC susceptibility, with higher GG genotype and G allele distribution in HCC patients than in non-HCC subjects. Such association was not detected in HCV patients compared to controls. EpCAM gene expression levels, determined in blood by RT-qPCR, and its serum protein expression levels, determined by ELISA, were significantly higher in GG relative to GC+CC genotype carriers in HCV and HCC patients in a recessive model. ROC analysis of EpCAM protein levels revealed significant discriminatory power between HCC patients and non-HCC subjects, with improved diagnostic accuracy when combining α-fetoprotein and EpCAM compared to that of α-fetoprotein alone. Altogether, EpCAM rs62139665 polymorphism is significantly associated with HCC and with EpCAM gene and protein expression levels in the Egyptian population. Moreover, serum EpCAM levels may hold promise for HCC diagnosis and for improving the diagnostic accuracy of α-fetoprotein.

2021
El-Fattah, A. A. A., N. A. H. Sadik, O. G. Shaker, A. M. Kamal, and N. N. Shahin, "Serum Long Non-Coding RNAs PVT1, HOTAIR, and NEAT1 as Potential Biomarkers in Egyptian Women with Breast Cancer.", Biomolecules, vol. 11, issue 2, 2021. Abstract

Long non-coding RNAs play an important role in tumor growth, angiogenesis, and metastasis in several types of cancer. However, the clinical significance of using lncRNAs as biomarkers for breast cancer diagnosis and prognosis is still poorly investigated. In this study, we analyzed the serum expression levels of lncRNAs PVT1, HOTAIR, NEAT1, and MALAT1, and their associated proteins, PAI-1, and OPN, in breast cancer patients compared to fibroadenoma patients and healthy subjects. Using quantitative real-time PCR (qRT-PCR), we compared the serum expression levels of the four circulating lncRNAs in patients with breast cancer ( = 50), fibroadenoma ( = 25), and healthy controls ( = 25). The serum levels of PAI-1 and OPN were measured using ELISA. Receiveroperating-characteristic (ROC) analysis and multivariate logistic regression were used to evaluate the diagnostic value of the selected parameters. The serum levels of HOTAIR, PAI-1, and OPN were significantly higher in breast cancer patients compared to controls and fibroadenoma patients. The serum level of PVT1 was significantly higher in breast cancer patients than in the controls, while that of NEAT1 was significantly lower in breast cancer patients compared to controls and fibroadenoma patients. Both ROC and multivariate logistic regression analyses revealed that PAI-1 has the greatest power in discriminating breast cancer from the control, whereas HOTAIR, PAI-1, and OPN have the greatest power in discriminating breast cancer from fibroadenoma patients. In conclusion, our data suggest that the serum levels of PVT1, HOTAIR, NEAT1, PAI-1, and OPN could serve as promising diagnostic biomarkers for breast cancer.

Arab, H. H., M. M. Safar, and N. N. Shahin, "Targeting ROS-Dependent AKT/GSK-3β/NF-κB and DJ-1/Nrf2 Pathways by Dapagliflozin Attenuates Neuronal Injury and Motor Dysfunction in Rotenone-Induced Parkinson's Disease Rat Model.", ACS chemical neuroscience, vol. 12, issue 4, pp. 689-703, 2021. Abstract

Dapagliflozin, a selective sodium-glucose co-transporter 2 (SGLT2) inhibitor, has emerged as a promising neuroprotective agent in murine models of epilepsy and obesity-induced cognitive impairment through its marked antioxidant/antiapoptotic features. However, the impact of dapagliflozin on the pathogenesis of Parkinson's disease (PD) is lacking. Hence, the present study aimed at exploring the potential neuroprotective effects of dapagliflozin against PD-associated neurodegenerative aberrations/motor dysfunction in rotenone-induced PD rat model. Rotenone (1.5 mg/kg) was subcutaneously administered every other day for 3 weeks. The expression of target signals was investigated using qPCR, Western blotting, ELISA, and immunohistochemistry. Dapagliflozin (1 (mg/kg)/day, by gavage for 3 weeks) attenuated PD motor dysfunction and improved motor coordination in the open-field and rotarod tests without triggering hypoglycemia. It also diminished the histopathologic alterations and α-synuclein expression and augmented tyrosine hydroxylase and dopamine levels. Dapagliflozin markedly alleviated neuronal oxidative stress via lowering lipid peroxides with consequent restoration of the disturbed DJ-1/Nrf2 pathway. Moreover, dapagliflozin counteracted ROS-dependent neuronal apoptosis and upregulated GDNF and its downstream PI3K/AKT/GSK-3β (Ser9) pathway. Meanwhile, it suppressed neuroinflammation via curbing the activation of NF-κB pathway and TNF-α levels. Together, these pleiotropic neuroprotective effects highlight the promising role of dapagliflozin in the management of PD.

Hanafy, M. M., J. Z. Lindeque, S. A. EL-Maraghy, A. - H. Z. Abdel-Hamid, and N. N. Shahin, "Time-based investigation of urinary metabolic markers for Type 2 diabetes: Metabolomics approach for diabetes management.", BioFactors (Oxford, England), 2021. Abstract

Diabetes is considered one of the most important health emergencies worldwide and Egypt has 8.2 million diabetic patients according to the International Diabetes Federation report in 2017. The objective of this study was to monitor the time-course variation in the metabolic profile of diabetic rats to detect urinary metabolic biomarkers using the metabolomics approach. Type 2 diabetes was induced in male Wistar albino rats using a single intraperitoneal injection of 40 mg/kg of streptozotocin following oral administration of 10% fructose in drinking water for 3 weeks. Then, urine was collected for 24 h from rats at three time points (0, 2, and 4 weeks after confirmation of diabetes), and were analyzed by nuclear magnetic resonance (H -NMR), followed by multivariate data analysis. The results from H -NMR pointed out that d-glucose, taurine, l-carnitine, l-fucose, 1,5-anhydrosorbitol, and d-galactose levels showed consistent significant variation (p < 0.05) between the positive (diabetic) and negative (normal) controls during the whole experimental period. Also, with the disease progression, myoinositol, and l-phenylalanine levels were significantly altered (p < 0.05) after 2 weeks and this alteration was maintained till the end of the 4-week experimental period in the positive control group. From the results of the present study, it could be concluded that we cannot depend only on glucose levels for prognostic purposes since there are other metabolic disturbances in diabetes which need to be tracked for better disease prognosis.

2020
Motawi, T. M. K., Z. M. Abdel-Nasser, and N. N. Shahin, "Ameliorative Effect of Necrosulfonamide in a Rat Model of Alzheimer's Disease: Targeting Mixed Lineage Kinase Domain-like Protein-Mediated Necroptosis.", ACS chemical neuroscience, vol. 11, issue 20, pp. 3386-3397, 2020. Abstract

Alzheimer's disease (AD) is a progressively debilitating neurodegenerative disorder that has no effective remedy, so far, with available therapeutic modalities being only symptomatic and of modest efficacy. Necroptosis is a form of controlled cell death with a recently emerging link to the pathogenesis of several neurodegenerative diseases. This study investigated the role of necroptosis in the pathogenesis of AD and evaluated the potential beneficial effect of the necroptosis inhibitor, necrosulfonamide (NSA), in a rat model of AD. AD was induced by oral administration of AlCl (17 mg/kg/day) for 6 consecutive weeks. Administration of NSA (1.65 mg/kg/day) intraperitoneally for 6 weeks significantly amended AlCl-induced spatial learning and memory deficits, as demonstrated by enhanced rat performance in Morris water and Y-mazes. NSA alleviated the abnormally high hippocampal expression of tumor necrosis factor-alpha (TNF-α), β-site amyloid precursor protein cleaving enzyme 1 (BACE1), β-amyloid, glycogen synthase kinase-3β (GSK-3β), phosphorylated tau protein, and acetylcholinesterase with concordant replenishment of acetylcholine. The amendments of AD perturbations achieved by NSA correlated with its inhibitory effect on the phosphorylation of the key necroptotic executioner, mixed lineage kinase domain-like protein (MLKL). Histopathological alterations supported the biochemical findings. In conclusion, NSA treatment represents a promising anti-Alzheimer's approach, mitigating AD neuropathologies via targeting MLKL-dependent necroptosis.

Motawi, T. K., N. N. Shahin, K. Awad, A. S. Maghraby, D. N. Abd-Elshafy, and M. M. Bahgat, "Glycolytic and immunological alterations in human U937 monocytes in response to H1N1 infection.", IUBMB life, vol. 72, issue 11, pp. 2481-2498, 2020. Abstract

We monitored changes that took place in glycolytic enzymes, the pyruvate end product of glycolysis, tumor necrosis factor α (TNFα), and toll-like receptors (TLRs) both at the transcriptional and translational levels upon direct interaction between PR8-H1N1 and the human monocytes U937 in vitro system. U937 were first treated with H1N1 infectious viral particles or phorbol-12-myristate-13-acetate (PMA) or left untreated and later infected with the H1N1 virus. Levels of phosphofructokinase 1 (PFK1) and pyruvate were biochemically quantified. In addition, levels of TNFα, TLR3, and TLR7 were measured by ELISA. The transcriptional profiles of PFKs, inflammatory cytokines, TLR3 and TLR7 were relatively quantified by qRT-PCR. The results generally revealed significant changes in both the transcriptional and translational profiles of the studied biochemical and immunological parameters upon influenza infection in a time-dependent manner. In conclusion, H1N1 infection triggers transcriptional and translational changes in immortalized human monocytes, which might serve as markers for infection subject for further validation for their specificities.

Motawi, T. K., N. N. Shahin, A. S. Maghraby, M. Kirschfink, D. N. Abd-Elshafy, K. Awad, and M. M. Bahgat, "H1N1 Infection Reduces Glucose Level in Human U937 Monocytes Culture.", Viral immunology, vol. 33, issue 5, pp. 384-390, 2020. Abstract

Infection with influenza A (H1N1) virus contributes significantly to the global burden of acute respiratory diseases. Glucose uptake and metabolic changes are reported in different cell types after infections with different virus types, including influenza A virus. Alteration of glucose metabolism specifically in immune cells has major health consequences. The aim of this study was to monitor glucose concentration in unstimulated and stimulated U937 human monocytes with infectious or heat inactivated H1N1 or or in nonpathogenically stimulated monocytes with phorbol-12-myristate-13-acetate. Stimulated or unstimulated U937 human monocytes were subjected to H1N1 infection for different time points and the glucose profile in the growth medium was measured post infection. Results showed that regardless to whether the initial stimuli on U937 cells were of pathogen or nonpathogen origins, challenge infection by H1N1 causes a significant reduction of glucose levels 36 h post infection. In conclusion, H1N1 infection has a direct effect on the glucose uptake of U937 cells . This effect can be related to either H1N1 infection or cell differentiation status that might occur due to the exerted stimuli.

Shahin, N. N., G. T. Abd-Elwahab, A. A. Tawfiq, and H. M. Abdelgawad, "Potential role of aryl hydrocarbon receptor signaling in childhood obesity.", Biochimica et biophysica acta. Molecular and cell biology of lipids, vol. 1865, issue 8, pp. 158714, 2020. Abstract

BACKGROUND: There is a growing concern that junk food has contributed to the childhood obesity epidemic. Recently, experimental studies suggested that the aryl hydrocarbon receptor (AHR) gene is strongly linked to western diet-induced obesity.

AIM: This study investigated the potential role of AHR signaling in childhood obesity and the possible associations of the AHR-aryl hydrocarbon receptor repressor (AHRR)-cytochrome P450 1B1 (CYP1B1) axis with fatty acid homeostasis and the appetite-related hormones, leptin and ghrelin.

SUBJECTS AND METHODS: The study included 80 children; 54 obese and 26 non-obese of matched age and sex. Demographic data, anthropometric measurements, and lipid profile were assessed. Expression of AHR signaling genes was analyzed in blood cells by qRT-PCR. Serum insulin, leptin and ghrelin levels were measured using ELISA.

RESULTS: The statistical power of this study, calculated using G*Power version 3.1.9.2, was 90% (α = 0.05). AHR and CYP1B1 gene expression levels were upregulated in the obese group compared to controls, whereas AHRR, stearoyl-CoA desaturase 1 (SCD1), and peroxisome proliferator-activated receptor-γ (PPARγ) were downregulated. Serum leptin correlated positively, while serum ghrelin correlated negatively with both AHR and CYP1B1. Stratification of obese children by age revealed more activated AHR signaling in younger than in older children. Receiver-operating-characteristic (ROC) analysis revealed that AHR, AHRR and CYP1B1 could discriminate between obese and normal weight children. Multivariate analysis showed that AHRR, CYP1B1 and ghrelin could be significant independent predictors of obesity.

CONCLUSION: This study provides new insights into the molecular mechanisms contributing to childhood obesity by revealing alterations in the AHR-AHRR-CYP1B1 axis, which could serve as a promising therapeutic target for childhood obesity.

Abo-Elfadl, M. T., A. M. Gamal-Eldeen, M. F. Ismail, and N. N. Shahin, "Silencing of the cytokine receptor TNFRSF13B: A new therapeutic target for triple-negative breast cancer.", Cytokine, vol. 125, pp. 154790, 2020. Abstract

BACKGROUND: TNFRSF13B, TACI, is a member of the TNF receptor superfamily; it plays a key role in cancer cell proliferation and progression.

METHOD: Influence of silencing of human cytokine receptors on cell viability was screened by Luminescent Cell Viability Assay, after transfection of the siRNA library to find the maximum cell death superhits in both triple-negative MDA-MB-231 and double-positive MCF7 breast cells. The mode of cell death was investigated by dual DNA fluorescence staining. The expression of mRNAs of TACI, BAFF, BAFF-R, and APRIL was explored by qPCR. Immunocytofluorescence analysis was used to evaluate changes in TACI, Bcl-2, TNFR2, cyclin-D2, and PCNA. NF-kB p65, cell cycle, and necrosis/apoptosis (late and early) were analyzed by flow cytometry.

RESULTS: TACI is the most potent cytotoxic superhit resulted from high-throughput screening of the siRNA library, in both types of cells. Our findings indicated that silencing receptor TACI in both types of breast cancer cells led to significant cell death, after different intervals from siRNA transfection. Cell death mediators (TNFR2, Bcl-2, and NF-κB) were significantly decreased after TACI silencing. The key factors for cell division (Cyclin-D2 and PCNA) were significantly increased in silenced cells of both types but the cell cycle was arrested before the completion of mitosis. Expression of BAFF, BAFF-R and APRIL mRNA in TACI-silenced cells showed significant upregulation in MDA-MB-231 cells, while only BAFF-R and APRIL showed significant downregulation in MCF7 cells.

CONCLUSION: TACI silencing can be a new and promising therapeutic target for mesenchymal-stem like triple-negative breast cancer subtype.

Safar, M. M., N. N. Shahin, A. F. Mohamed, and N. F. Abdelkader, "Suppression of BACE1 and amyloidogenic/RAGE axis by sitagliptin ameliorates PTZ kindling-induced cognitive deficits in rats.", Chemico-biological interactions, vol. 328, pp. 109144, 2020. Abstract

The debilitating nature of cognitive impairment in epilepsy and the potential of some traditional antiepileptics to further deteriorate cognitive function are areas of growing concern. Glucagon-like peptide-1 (GLP-1) deficiency has been linked to reduced seizure threshold as well as cognitive dysfunction. Here, we tested whether sitagliptin (SITA), by virtue of its neuroprotective properties, could alleviate both epilepsy and associated cognitive dysfunction in a rat model of kindling epilepsy. Chemical kindling was induced by subconvulsive doses of pentylenetetrazol (PTZ) (30 mg/kg; i.p). SITA (50 mg/kg; p.o) was administered 1 h before PTZ injections. SITA conceivably attenuated PTZ hippocampal histological insult, preserved neuronal integrity and amended neurotransmitter perturbations in rat hippocampi paralleled with enhanced hippocampal GLP-1 levels as well as the downstream cAMP content and protein kinase A (PKA) activity. Moreover, SITA improved cognitive functioning of rats in the Morris water maze which was coupled with hampered hippocampal p(Ser)-tau and β-amyloid proteins. SITA replenished p(Ser)-glycogen synthase kinase-3β (GSK-3β). It also opposed the boosted matrix metalloproteinase-9 (MMP-9), brain-derived neurotrophic factor (BDNF), and insulin-like growth factor-1 (IGF-1) levels associated with PTZ administration along with mitigation of both β-secretase-1 (BACE1) immunoreactivity and receptor for advanced glycation end products (RAGE) protein level in rat hippocampi. In conclusion, SITA subdues epileptic and cognitive upshots of PTZ kindling in rats, which might correspond to the modulation of BACE1, amyloidogenic/RAGE axis as well as GSK-3β/MMP-9/BDNF signaling cascade. SITA effects are probably mediated via boosting GLP-1 and subsequently enhancing GLP-1/GLP-1R signaling.

2019
Abdelmonem, M., N. N. Shahin, L. A. Rashed, H. A. A. Amin, A. A. Shamaa, and A. A. Shaheen, "Hydrogen sulfide enhances the effectiveness of mesenchymal stem cell therapy in rats with heart failure: In vitro preconditioning versus in vivo co-delivery.", Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, vol. 112, pp. 108584, 2019 Feb 18. Abstract

Stem cell therapy represents a promising therapeutic avenue for cardiac disorders, including heart failure. Although stem cell transplantation showed encouraging preliminary results, the outcomes of clinical studies are still unsatisfactory. This study aimed to compare the outcomes of two therapeutic approaches, in vivo co-delivery of sodium hydrogen sulfide (NaHS) concomitant with bone marrow-derived mesenchymal stem cell (BMSC) transplantation and in vitro preconditioning of BMSCs with NaHS, both of which are intended to promote the success of stem cell therapy in rats with isoprenaline-induced heart failure. Heart failure developed 4 weeks after the subcutaneous injection of isoprenaline (170 mg/kg) for 4 consecutive days. The in vivo approach involved the co-delivery of intraperitoneally administered NaHS concomitant with BMSC transplantation for a period of 14 days. The in vitro approach involved preconditioning BMSCs with NaHS for 30 min before transplantation. Compared to treatment with BMSCs alone, in vitro preconditioning of BMSCs with NaHS improved left ventricular function as measured by echocardiography and electrocardiography and enhanced stem cell homing, proliferation and differentiation as manifested by higher cardiac expression of GATA-4 and myocyte enhancer factor 2. Moreover, the measurement of cardiac transforming growth factor beta 1 levels and histopathological investigation revealed mitigated fibrosis and myocardial injury scores. Compared with BMSC therapy alone, the in vivo approach enhanced stem cell homing and differentiation, alleviated fibrosis and augmented vascular endothelial growth factor (VEGF) and endothelial nitric oxide synthase (eNOS) expression. In conclusion, NaHS can potentiate the efficiency of BMSC therapy for heart failure by in vitro preconditioning or in vivo co-delivery. The in vitro approach is superior with regard to improving cardiac function in addition to enhancing stem cell proliferation, while the in vivo approach is superior with regard to increasing cardiac VEGF and eNOS expression.

Shahin, N. N., N. A. El-Nabarawy, A. S. Gouda, and B. Mégarbane, "The protective role of spermine against male reproductive aberrations induced by exposure to electromagnetic field - An experimental investigation in the rat.", Toxicology and applied pharmacology, vol. 370, pp. 117-130, 2019. Abstract

The exponentially increasing use of electromagnetic field (EMF)-emitting devices imposes substantial health burden on modern societies with particular concerns of male infertility. Limited studies have addressed the modulation of this risk by protective agents. We investigated the hazardous effects of rat exposure to EMF (900 MHz, 2 h/day for 8 weeks) on male fertility and evaluated the possible protective effect of the polyamine, spermine, against EMF-induced alterations. Exposure to EMF significantly decreased sperm count, viability and motility, and increased sperm deformities. EMF-exposed rats exhibited significant reductions in serum inhibin B and testosterone along with elevated activin A, follicle-stimulating hormone, luteinizing hormone and estradiol concentrations. Testicular steroidogenic acute regulatory protein (StAR), c-kit mRNA expression and testicular activities of the key androgenic enzymes 3β- and 17β-hydroxysteroid dehydrogenases were significantly attenuated following exposure to EMF. Exposure led to testicular lipid peroxidation, decreased catalase and glutathione peroxidase activities and triggered nuclear factor-kappa B p65, inducible nitric oxide synthase, cyclooxygenase-2 and caspase-3 overexpression. EMF-exposed rats showed testicular DNA damage as indicated by elevated comet parameters. Spermine administration (2.5 mg/Kg/day intraperitoneally for 8 weeks) prevented EMF-induced alterations in the sperm and hormone profiles, StAR and c-kit expression and androgenic enzyme activities. Spermine hampered EMF-induced oxidative, inflammatory, apoptotic and DNA perturbations. Histological and histomorphometric analysis of the testes supported all biochemical findings. In conclusion, rat exposure to EMF disrupts sperm and hormone profiles with underlying impairment of steroidogenesis and spermatogenesis. Spermine confers protection against EMF-associated testicular and reproductive aberrations, at least in part, via antioxidant, anti-inflammatory and anti-apoptotic mechanisms.

Motawi, T. M. K., N. A. H. Sadik, D. Sabry, N. N. Shahin, and S. A. Fahim, "rs2267531, a promoter SNP within glypican-3 gene in the X chromosome, is associated with hepatocellular carcinoma in Egyptians.", Scientific reports, vol. 9, issue 1, pp. 6868, 2019. Abstract

Hepatocellular carcinoma (HCC) is a major health concern in Egypt owing to the high prevalence of hepatitis C virus (HCV) infection. HCC incidence is characterized by obvious male predominance, yet the molecular mechanisms behind this gender bias are still unidentified. Functional variations in X-linked genes have more impact on males than females. Glypican-3 (GPC3) gene, located in the Xq26 region, has lately emerged as being potentially implicated in hepatocellular carcinogenesis. The current study was designed to examine the association of -784 G/C single nucleotide polymorphism (SNP) in GPC3 promoter region (rs2267531) with HCC susceptibility in male and female Egyptian HCV patients. Our results revealed a significant association between GPC3 and HCC risk in both males and females, evidenced by higher C allele and CC/C genotype frequencies in HCC patients when compared to controls. However, no such association was found when comparing HCV patients to controls. Moreover, GPC3 gene and protein expression levels were significantly higher in CC/C than in GG/G genotype carriers in males and females. The CC/C genotype exhibited a significant shorter overall survival than GG/G genotype in HCC patients. In conclusion, GPC3 rs2267531 on the X chromosome is significantly associated with HCC, but not with HCV infection, in the Egyptian population.

Ali, S. O., N. N. Shahin, M. M. Safar, and S. M. Rizk, "Therapeutic potential of endothelial progenitor cells in a rat model of epilepsy: Role of autophagy", Journal of Advanced Research, vol. 18, pp. 101 - 112, 2019. AbstractWebsite

Epilepsy is one of the most well-known neurological conditions worldwide. One-third of adult epileptic patients do not respond to antiepileptic drugs or surgical treatment and therefore suffer from the resistant type of epilepsy. Stem cells have been given substantial consideration in the field of epilepsy therapeutics. The implication of pathologic vascular response in sustained seizures and the eminent role of endothelial progenitor cells (EPCs) in maintaining vascular integrity tempted us to investigate the potential therapeutic effects of EPCs in a pentylenetetrazole (PTZ)-induced rat model of epilepsy. Modulation of autophagy, a process that enables neurons to maintain an equilibrium of synthesis, degradation and subsequent reprocessing of cellular components, has been targeted. Intravenously administered EPCs homed into the hippocampus and amended the deficits in memory and locomotor activity. The cells mitigated neurological damage and the associated histopathological alterations and boosted the expression of brain-derived neurotrophic factor. EPCs corrected the perturbations in neurotransmitter activity and enhanced the expression of the downregulated autophagy proteins light chain protein-3 (LC-3), beclin-1, and autophagy-related gene-7 (ATG-7). Generally, these effects were comparable to those achieved by the reference antiepileptic drug, valproic acid. In conclusion, EPCs may confer therapeutic effects against epilepsy and its associated behavioural and biochemical abnormalities at least in part via the upregulation of autophagy. The study warrants further research in experimental and clinical settings to verify the prospect of using EPCs as a valid therapeutic strategy in patients with epilepsy.

2018
Shahin, N. N., N. F. Abdelkader, and M. M. Safar, "A Novel Role of Irbesartan in Gastroprotection against Indomethacin-Induced Gastric Injury in Rats: Targeting DDAH/ADMA and EGFR/ERK Signaling.", Scientific reports, vol. 8, issue 1, pp. 4280, 2018 Mar 09. Abstract

The advent of angiotensin II type 1 receptor blockers (ARBs) as intriguing gastroprotective candidates and the superior pharmacokinetics and pharmacodynamics displayed by irbesartan compared to many other ARBs raised the interest to investigate its gastroprotective potential in a rat model of gastric injury. Irbesartan (50 mg/Kg) was orally administered to male Wistar rats once daily for 14 days; thereafter gastric injury was induced by indomethacin (60 mg/Kg, p.o). Irbesartan reduced gastric ulcer index, gastric acidity, and ameliorated indomethacin-induced gastric mucosal apoptotic and inflammatory aberrations, as demonstrated by hampering caspase-3, prostaglandin E and tumor necrosis factor-alpha levels and cyclooxygenase-2 mRNA expression. This ARB increased mucosal dimethylarginine dimethylaminohydrolase-1 (DDAH-1) gene expression and decreased elevated levels of matrix metalloproteinase-9, asymmetric dimethylarginine (ADMA), epidermal growth factor receptor (EGFR) mRNA and phosphorylated extracellular signal-regulated kinase 1 and 2 (pERK1/2). Histopathological evaluation corroborated biochemical findings. Overall efficacy of irbesartan was comparable to ranitidine, the widely used H receptor blocker. In conclusion, irbesartan exerts significant gastroprotection against indomethacin-induced mucosal damage via acid-inhibitory, anti-inflammatory, anti-apoptotic and extracellular matrix remodeling mechanisms that are probably mediated, at least partly, by down-regulating DDAH/ADMA and EGFR/ERK1/2 signaling.

Motawi, T. K., M. R. Mohamed, N. N. Shahin, M. A. M. Ali, and M. A. Azzam, "Time-course expression profile and diagnostic potential of a miRNA panel in exosomes and total serum in acute liver injury.", The international journal of biochemistry & cell biology, vol. 100, pp. 11-21, 2018 Jul. Abstract

Circulating miRNAs have recently emerged as attractive candidates for biomarker discovery. However, they have a variant distribution in circulation, and the diagnostic significance of their compartmentalization is yet to be elucidated. This study explored the time-course expression profile and the diagnostic potential of miRNAs-122a-5p, 192-5p, 193a-3p and 194-5p in exosomal and total serum compartments in two rat models of acute liver injury (ALI). Exosomes were isolated and characterized in terms of morphology, size and CD-63 surface marker expression. Exosomal, serum and hepatic miRNAs were quantified using q-RT-PCR. An inverse expression pattern of hepatic and total serum miRNAs was observed following acetaminophen or thioacetamide-induced liver injury. Conversely, exosomal miRNAs expression pattern varied according to the type of injury. Overall, ROC analysis revealed superior discriminatory ability of exosomal miRNA-122a-5p following either acetaminophen or thioacetamide injury with earlier diagnostic potential and a wider diagnostic window compared to the corresponding total serum counterpart. Moreover, exosomal miRNAs showed higher correlation with ALT activity in both models. In conclusion, exosomal miRNA-122a-5p shows higher diagnostic performance with a broader diagnostic time window and an earlier diagnostic potential than its serum counterpart in ALI. Furthermore, exosomal miRNAs-122a-5p, 192-5p and 193a-3p exhibit an injury-specific signature in ALI and can be used not only as diagnostic tools in liver injury but also to differentiate between different etiologies of injury.

2017
Shahin, N. N., and M. M. Mohamed, "Nano-sized titanium dioxide toxicity in rat prostate and testis: Possible ameliorative effect of morin.", Toxicology and applied pharmacology, vol. 334, pp. 129-141, 2017 11 01. Abstract

This study investigated the effect of short-term oral exposure to nano-sized titanium dioxide (nTiO2) on Wistar rat prostate and testis, and the associating reproductive-related alterations. The study also evaluated the potential ameliorative effect of the natural flavonoid, morin, on nTiO2-induced aberrations. Intragastric administration of nTiO2 (50mg/kg/day for 1, 2 and 3weeks) increased testicular gamma-glutamyltransferase (γ-GT) activity and decreased testicular steroidogenic acute regulatory protein (StAR) and c-kit gene expression, serum testosterone level and sperm count. nTiO2-treated rats also exhibited prostatic and testicular altered glutathione levels, elevated TNF-α levels, up-regulated Fas, Bax and caspase-3 gene expression, down-regulated Bcl-2 gene expression and enhanced prostatic lipid peroxidation. Sperm malformation and elevated testicular acid phosphatase (ACP) activity and malondialdehyde level, serum prostatic acid phosphatase activity, prostate specific antigen (PSA), gonadotrophin and estradiol levels occurred after the 2 and 3week regimens. Morin (30mg/kg/day administered intragastrically for 5weeks) mitigated nTiO2-induced prostatic and testicular injury as evidenced by lowering serum PSA level, testicular γ-GT and ACP activities and TNF-α level, along with hampering both intrinsic and extrinsic apoptotic pathways. Moreover, morin alleviated prostatic lipid peroxidation, raised prostatic glutathione level, and relieved testicular reductive stress. Additionally, morin increased testicular StAR and c-kit mRNA expression, raised the sperm count, reduced sperm deformities and modified the altered hormone profile. Histopathological evaluation supported the biochemical findings. In conclusion, morin could ameliorate nTiO2-induced prostatic and testicular injury and the corresponding reproductive-related aberrations via redox regulatory, anti-inflammatory and anti-apoptotic mechanisms, promoting steroidogenesis and spermatogenesis, and improving sperm count and morphology.

2016
Motawi, T. K., O. G. Shaker, N. N. Shahin, and N. M. Ahmed, "Angiotensin-converting enzyme insertion/deletion polymorphism association with obesity and some related disorders in Egyptian females: a case-control observational study.", Nutrition & metabolism, vol. 13, pp. 68, 2016. Abstract

BACKGROUND: According to the WHO report in 2015, obesity is the fifth leading cause of death worldwide, and the prevalence of Egyptian female obesity is 37.5 %. Since obesity is highly influenced by genetics, and adipose tissue renin-angiotensin system is over-activated in obesity, the effect of angiotensin-converting enzyme (ACE) insertion/deletion (I/D) polymorphism on obesity and related disorders was studied in several populations, because of its effect on ACE activity. Our objective was to study the association of ACE I/D polymorphism with obesity and certain related disorders, namely hypertension, insulin resistance and metabolic syndrome, in Egyptian females.

METHODS: Eighty female volunteers were recruited, blood pressure and body measurements were recorded and a fasting blood sample was obtained for the quantitation of glucose, lipid profile, insulin, leptin and identification of ACE I/D polymorphs. Subjects were grouped based on hypertension and obesity states. Comparisons of continuous parameters were made with independent sample t-test between two groups. The frequencies of ACE genotypes and alleles, and the association between gene polymorphism and metabolic parameters were assessed using chi-square or Fisher's exact test.

RESULTS: Genotype frequencies were in Hardy-Weinberg equilibrium for all groups. Genotype distribution did not differ significantly between controls and cases of all the studied disorders. Although DD carriers had apparently higher parameters of blood pressure, lipid profile and insulin resistance, only diastolic blood pressure was almost significant (p = 0.057). I-carriers were significantly less susceptible to hypertension than DD carriers having normal waist/hip ratio (p = 0.007, OR = 17.29, CI = 1.81-164.96) and normal conicity index (p = 0.024, OR = 7.00, CI = 1.36-35.93). In DD genotype carriers, a significant association was found between insulin resistance and high body mass index (p = 0.004, OR = 8.89, CI = 1.94-40.71), waist circumference (p = 0.003, OR = 9.63, CI = 2.14-43.36) and waist/height ratio (p = 0.034, OR = 6.86, CI = 1.25-37.61), although the variations in percentages between DD and I-carriers were not high enough to conclude an effect of ACE I/D on such an association.

CONCLUSIONS: In this sample of Egyptian females, ACE I/D polymorphism was not significantly associated with obesity nor with any of its related disorders studied. The I allele seemed protective against hypertension in subjects with normal, not high, waist/hip ratio and conicity index compared to DD genotype carriers.

Rizk, S. M., N. N. Shahin, and O. G. Shaker, "Association between SIRT1 Gene Polymorphisms and Breast Cancer in Egyptians.", PloS one, vol. 11, issue 3, pp. e0151901, 2016. Abstract

BACKGROUND: Breast cancer is reported to cause the highest mortality among female cancer patients. Previous studies have explored the association of silent mating-type information regulator 2 homolog 1 (SIRT1) gene expression with prognosis in breast cancer. However, no studies exist, so far, on the role of SIRT1 gene polymorphism in breast cancer risk or prognosis. The present study aimed to assess the association between SIRT1 gene polymorphisms and breast cancer in Egyptians.

METHODS: The study comprised 980 Egyptian females divided into a breast cancer group (541 patients) and a healthy control group (439 subjects). SIRT1 gene single nucleotide polymorphisms (SNPs) rs3758391, rs3740051 and rs12778366 were genotyped using real-time polymerase chain reaction (RT-PCR). Allelic and genotypic frequencies were determined in both groups and association with breast cancer and clinicopathological characteristics was assessed.

RESULTS: Breast cancer patients exhibited elevated serum SIRT1 levels which varied among different tumor grades. SIRT1 rs3758391 and rs12778366 TT genotypes were more frequent, exhibited higher SIRT1 levels than CC and CT genotypes and were associated with histologic grade and lymph node status. SIRT1 rs12778366 TT genotype also correlated with negative estrogen receptor (ER) and progesterone receptor (PR) statuses. The T allele frequency for both SNPs was higher in breast cancer patients than in normal subjects. Combined GG and AG genotypes of rs3740051 were more frequent, showed higher serum SIRT1 levels than the AA genotype, and were associated with ER and PR expression. Furthermore, inheritance of the G allele was associated with breast cancer.

CONCLUSIONS: Our findings reveal that rs3758391 and rs12778366 polymorphisms of SIRT1 gene are associated with breast cancer risk and prognosis in the Egyptian population.

2015
EL-Maraghy, S. A., S. M. Rizk, and N. N. Shahin, "Gastroprotective effect of crocin in ethanol-induced gastric injury in rats", Chemico-biological interactions, vol. 229: Elsevier, pp. 26–35, 2015. Abstract

The present study investigated the gastroprotective effect of crocin in ethanol-induced gastric injury in rats. Rats were allocated into a normal group, an ulcer group, a crocin-treated group, an ulcer group pretreated with crocin, and an ulcer group pretreated with omeprazole as a reference anti-ulcer drug. Rats were sacrificed 3 hours after ethanol administration. Prophylactic administration of crocin (50 mg/Kg/day, i.p.) for 3 consecutive days before the administration of 70% ethanol (10 ml/Kg, orally) resulted in significant gastroprotection compared to ethanol-ulcerated rats as manifested by significant reduction in the gastric ulcer index. Crocin pretreatment increased ethanol-lowered levels of gastric juice mucin and mucosal prostaglandin E2 (PGE2) and interleukin-6. Moreover, crocin significantly decreased ethanol-elevated tumor necrosis factor-alpha level, myeloperoxidase activity and heat shock protein 70 mRNA and protein levels. It also restored ethanol-altered mucosal levels of glutathione, malondialdehyde and superoxide dismutase activity. Furthermore, crocin-pretreatment alleviated ethanol-induced mucosal apoptosis as revealed by significant down-regulation of cytochrome c and caspase-3 mRNA expression, significant decrease in caspase-3 activity and mitigated DNA fragmentation as indicated by significant decrements in comet parameters. The protective efficacy of crocin was further supported by histological assessment. No significant difference was observed between crocin and omeprazole (20 mg/Kg orally 1 hour before ethanol administration) regarding their mucin-secretagogue and antioxidant effects, as well as their effects on TNF-α, IL-6 and cytochrome c. On the other hand, omeprazole was superior in enhancing PGE2 level and in alleviating neutrophil infiltration, caspase-3 activation and DNA fragmentation. Conclusively, crocin protects rat gastric mucosa against ethanol-induced injury via anti-inflammatory, anti-oxidative, anti-apoptotic and mucin-secretagogue mechanisms that are probably mediated by enhanced PGE2 release.

2014
Abd El Tawab, A. M., N. N. Shahin, and M. M. AbdelMohsen, "Protective effect of Satureja montana extract on cyclophosphamide-induced testicular injury in rats", Chemico-Biological Interactions, vol. 224: Elsevier, pp. 196–205, 2014. Abstract

The present study investigated the protective effect of Satureja montana extract against cyclophosphamide-induced testicular injury in rats. Total phenolic and flavonoid contents of the extract were 1.03% and 0.34% w/w of dry herb expressed as chlorogenic acid and quercetin, respectively. HPLC analysis identified caffeic, syringic and rosmarinic acids as the chief phenolic acids, and rutin as the major flavonoid in the extract. Oral daily administration of S. montana extract (50 mg/kg/day) for 7 days before and 7 days after an intraperitoneal injection of cyclophosphamide (200 mg/kg) restored the reduced relative testicular weight, serum testosterone level and testicular alkaline phosphatase activity, raised the lowered testicular sorbitol dehydrogenase and acid phosphatase activities, and decreased the elevated testicular hemoglobin absorbance. It also attenuated lipid peroxidation, restored the lowered glutathione content, glucose-6-phosphate dehydrogenase, glutathione peroxidase and glutathione reductase activities, and improved total antioxidant capacity. Moreover, S. montana extract mitigated testicular DNA fragmentation, decreased the elevated Fas and Bax gene expression, up-regulated the decreased Bcl-2 and peroxisome proliferator-activated receptor-gamma (PPAR-c) gene expression and normalized Akt1 protein level. Histopathological investigation confirmed the protective effects of the extract. Conclusively, S. Montana extract protects the rat testis against cyclophosphamide-induced damage via anti-oxidative and anti-apoptotic mechanisms that seem to be mediated, at least in part, by PPAR-c and Akt1 up-regulation.

2013
Motawi, T., H. Abd-Elgawad, and N. Shahin, "Effect of protein malnutrition on the metabolism and toxicity of cisplatin, 5-fluorouracil and mitomycin C in rat stomach", Food and Chemical Toxicology, vol. 56: Elsevier, pp. 467–482, 2013. Abstract

This study investigated the effect of protein malnutrition on metabolism and toxicity of cisplatin (CP), 5-
fluorouracil (FU) and mitomycin C (MMC) in rat stomach. Weanling male Wistar rats received a normal
(24%) or low (2.5%) protein diet for 28 days and were allocated into: normally-f ed control, protein-malnourished
control (PM), 3 normally-fed drug-treated groups and 3 protein-malnourished drug-treated
groups (PM-CP, PM-FU and PM-MMC). Cisplatin and MMC were injected intraperitoneally (8 mg/kg on
day 26 and 1 mg/kg/day for 7 days, respectively). 5-Fluorour acil was given orally (50 mg/kg /day for
5 days). Compared with normally-fed counterparts, PM-CP rats exhibited higher glutathione S-transferase,
aminopeptidase N and cysteine S-conjugate beta-lyase (CCBL) and lower gamma-glutamyltr ansferase
activities, PM-FU rats exhibited decreased dihydropyrimidine dehydrogenase and cytochrome P450
1A1/2 activities and PM-MMC rats showed higher quinone reductase and depleted xanthin e oxidase
activities. Protein-malnourished drug-treate d groups exhibited exacerbated gastrotoxicity, relative to
normally-fed counterparts, manifested by lower mucus levels, higher permeability and histopathological
deterioration, along with increased oxidati ve stress in PM-CP rats and exaggerated prostaglandin E2 production
in PM-MMC rats. Conclusively, protein malnutrition alters CP, FU and MMC metabolism in rat
stomach by enhancing CCBL pathway for CP activa tion, delaying FU elimination and activating two-electron
reduction of MMC, potentiating their gast rotoxicity.

2008
Motawi, T., H. Abd Elgawad, and N. Shahin, "Gastroprotective effect of leptin in indomethacin-induced gastric injury", Journal of Biomedical Science, vol. 15, no. 3: Springer, pp. 405–412, 2008. Abstract

This study investigated the involvement of
neutrophil infiltration, disturbances in nitric oxide (NO)
generation and oxidative stress in indomethacin-induced
gastric ulcer, and the possible gastroprotective potentials of
leptin, known for its angiogenic effect. Male Wistar albino
rats (180–220 g) were allocated into a normal control
group, ulcer control group (received a single dose of
indomethacin 40 mg/kg p.o.) and an ulcer group pretreated
with leptin (10 lg/kg i.p. 30 min before ulcer induction).
The animals were killed 6 h after indomethacin administration
and their gastric juice, serum and mucosal tissue
were used for gastric injury evaluation. Indomethacin
produced multiple lesions in glandular mucosa, evidenced
by marked increase in gastric ulcer index (GUI) accompanied
by significant increases in gastric juice acidity,
tissue myeloperoxidase (MPO) activity, serum NO and
tissue conjugated diene (CD), and marked decreases in
tissue NO and glutathione (GSH) as well as glutathione
reductase (GR) and superoxide dismutase (SOD) activities,
while gastric juice mucin and tissue glutathione peroxidase
(GPx) were not affected. Leptin exerted significant gastroprotection
as evidenced by significantly decreased GUI
and attenuated neutrophil infiltration. Leptin significantly
increased mucin and tissue NO, restored GR and SOD
activities and up-regulated GPx activity. It failed to affect
acidity, serum NO, GSH and CD. These results suggest
that leptin confers significant gastroprotection against
indomethacin-induced injury through interfering with
neutrophil infiltration, NO production and oxidative stress.
Keywords Indomethacin  Gastric ulcer  Leptin 
Neutrophil infiltration  NO  Oxidative stress

2007
Motawi, T., H. Abd Elgawad, and N. Shahin, "Modulation of indomethacin-induced gastric injury by spermine and taurine in rats", Journal of Biochemical and Molecular Toxicology, vol. 21, no. 5: Wiley, pp. 280–288, 2007. Abstract

This study investigated the involvement
of neutrophil infiltration, nitric oxide (NO) generation,
and oxidative stress in indomethacin-induced ulcer
and the possible gastroprotective potentials of spermine
and taurine, known for their tissue regenerating
and antioxidant effects, respectively. Male Wistar albino
rats (180–220 g) were allocated into a normal control
group, ulcer control group (received a single dose
of indomethacin 40 mg/kg p.o.), and two ulcer groups
pretreated with spermine (150 mg/kg p.o. 1 h before
ulcer induction) and taurine (250 mg/kg i.p. for three
consecutive days before ulcer induction). The animals
were killed 6 h after indomethacin administration, and
the gastric juice, serum, and mucosal tissue were used
for gastric injury evaluation. Both modulators significantly
ameliorated the indomethacin–induced gastric
lesions in glandularmucosa. Notably, spermine exhibited
the most pronounced effect as manifested by great
reduction in the gastric ulcer index, normalization of
the elevated gastric acidity, and triggering of mucin
production. Spermine and taurine were able to decrease
the elevated levels of gastric myeloperoxidase,
conjugated diene, and serum NO. However, the lowered
tissue NO content was markedly elevated only
by taurine. The antioxidant action of taurine was illustrated
by restoration of the depressed content of
glutathione, normalization of the inhibited activities
of glutathione reductase, and superoxide dismutase.
These results suggest that spermine and taurine confer
significant gastroprotection against indomethacininduced
gastric injury with the priority of spermine.
C 
2007 Wiley Periodicals, Inc. J Biochem Mol Toxicol
21:280–288, 2007; Published online inWiley InterScience
(www.interscience.wiley.com). DOI 10:1002/jbt.20194
KEYWORDS: Indomethacin; Gastric Ulcer; Spermine;
Taurine; Neutrophil Infiltration; NO; Oxidative Stress