Publications by authors named "Noha M Mesbah"

37 Publications

Anti-cancer activity of two novel heterocyclic compounds through modulation of VEGFR and miR-122 in mice bearing Ehrlich ascites carcinoma.

Eur J Pharmacol 2021 Feb 21;892:173747. Epub 2020 Nov 21.

Department of Medicinal Chemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, 41522, Egypt; Center for Molecular Spectroscopy and Dynamic, Institute for Basic Science, Korea University, Seoul, 02841, Republic of Korea. Electronic address:

Metastasis in breast cancer is a leading cause of mortality among women in many countries. This study investigated the anti-cancer role of benzoimidazoquinazoline and benzimidazotriazin; two novel compounds that were designed, synthesized, structurally elucidated, and biologically evaluated as potent anti-angiogenic agents that act through inhibition of vascular endothelial growth factor receptor-2 (VEGFR2). Breast cancer was induced by inoculation of Ehrlich Ascites Carcinoma (EAC) cells. Seventy swiss albino mice were randomly divided into 7 groups, 10 animals each: (1) normal, (2) control EAC group, (3) cisplatin treated group, (4&5) benzoimidazoquinazoline treated (5 mg/kg and 10 mg/kg), (6&7) benzimidazotriazin treated (5 mg/kg and 10 mg/kg). The expression of miR-122 was assessed in the tumor tissue by quantitative PCR, and the VEGF level was determined in serum by ELISA. VEGFR2 and cluster of differentiation (CD)34 were assessed by immunohistochemistry. Serum ALT, AST, creatinine, and urea were measured. Treatment with benzoimidazoquinazoline and benzimidazotriazin decreased tumor weight and serum levels of VEGF, and down-regulated expression of VEGFR2 and CD34 in the tumor tissue. miR-122 was upregulated, particularly in the benzimidazotriazin (10 mg/kg) group. Relative to cisplatin, the novel compounds were less toxic to kidneys. Benzoimidazoquinazoline and benzimidazotriazin are promising anti-cancer agents that act through inhibition of angiogenesis and thus provide a new strategy for advancement of chemotherapy.
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http://dx.doi.org/10.1016/j.ejphar.2020.173747DOI Listing
February 2021

Ginger Extract Loaded into Chitosan Nanoparticles Enhances Cytotoxicity and Reduces Cardiotoxicity of Doxorubicin in Hepatocellular Carcinoma in Mice.

Nutr Cancer 2020 Sep 25:1-16. Epub 2020 Sep 25.

Department of Biochemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt.

This study aimed to investigate the impact of ginger extract (GE) loaded into chitosan nanoparticles (CNPs) in enhancing cytotoxicity and reducing cardiotoxicity of doxorubicin (DXN) in hepatocellular carcinoma (HCC) induced mice. DXN and GE were loaded into CNPs and cytotoxicity of loaded and unloaded drugs against HepG2 cells was evaluated. HCC was induced in male albino mice by injection of diethylnitrosamine (DINA). Mice were divided into eight groups ( = 15): (1) normal control, (2) DINA, (3) CNPs, (4) free DXN, (5) CNPs DXN, (6) free GE, (7) CNPs GE, and (8) CNPs DXN + CNPs GE. Both GE and DXN loaded into CNPs showed a greater decline in cell viability of HepG2 cells than the unloaded forms. GE CNPs displayed pronounced anticancer activity In Vivo through apoptosis, greater down-regulation of multidrug resistance 1, enhancement of anti-oxidant activity and depletion of vascular endothelial growth factor content in liver tissues. GE CNPs in combination with DXN CNPs showed nearly normal hepatic lobule architecture and the greatest increase in apoptotic cell count. Co-treatment group had decreased cardiac malondialdehyde, tumor necrosis factor-α and serum activity of creatine kinase and lactate dehydrogenase. Combination of GE CNPs and DXN CNPs might be a potentially effective therapeutic approach for HCC.
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http://dx.doi.org/10.1080/01635581.2020.1823436DOI Listing
September 2020

Long non-coding RNA MEG3 and its genetic variant rs941576 are associated with rheumatoid arthritis pathogenesis in Egyptian patients.

Arch Physiol Biochem 2020 Jul 1:1-8. Epub 2020 Jul 1.

Department of Biochemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt.

Rheumatoid arthritis (RA) is a joint destructive disorder. This study aimed to assess lncRNA MEG3 expression and its variant rs941576 in Egyptian patients with RA. 100 RA patients and 100 healthy individuals were enrolled in the study. Quantitative PCR was used for expression analysis and allelic discrimination technology for genotyping. LncRNA MEG3 was down-regulated in RA patients and negatively associated with RA clinical features and HIF-1α and VEGF serum levels. On the contrary, it was positively associated with BAX serum levels in RA patients. The major A allele of rs941576 variant was associated with RA patients ( = .0003). AA genotype showed a significant decrease in lncRNA MEG3 expression and BAX and increase in HIF-1α and VEGF. Serum lncRNA MEG3 expression showed negative association with increased susceptibility to RA. MEG3 gene rs941576 (A/G) polymorphism was associated with increased severity of RA in the current population.
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http://dx.doi.org/10.1080/13813455.2020.1784951DOI Listing
July 2020

Therapeutic effect of bone marrow mesenchymal stem cells in a rat model of carbon tetrachloride induced liver fibrosis.

Biomed J 2020 May 7. Epub 2020 May 7.

Department of Biochemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt.

Background: Liver fibrosis is a major medical problem with high mortality and morbidity rates where the formation of regenerative nodules and cirrhosis leads to loss of liver function and may result in the development of hepatocellular carcinoma. Bone marrow Mesenchymal stem cells (BM-MSCs) have drawn attention as a novel approach for treatment of liver fibrosis. This study aimed to evaluate the therapeutic effect of BM-MSCs on the liver structure in carbon tetrachloride (CCl) induced liver fibrosis in male rats relative to resveratrol and Silybum marianum as standard drugs derived from herbal plants.

Methods: Fifty adult male albino rats (Sprague Dawley strain; 180-220 g mean body weight) were purchased from the Laboratory Animal Unit in the Nile Center of Experimental Research, Mansoura, Egypt. Liver function were detrmined, isolation and preparation of BM- MSCs and detection of cell-surface markers by flow cytometry.

Results: Animals exposed to CCl developed liver injury characterized by significant increase of liver enzymes, malondialdehyde (MDA), tumor necrosis factor alpha (TNFα), and CYP450, inhibition of antioxidant enzymes, and decreased albumin. Treatment with stem cells enhanced liver state more effectively than resveratrol and Silybum marianum. It significantly decreased AST, ALT, ALP, MDA, TNF-α, and CYP450 and increased albumin, SOD, GSH, GST, and CAT. Histopathological study and atomic force microscope results confirmed the therapeutic effects of MSCs.

Conclusions: BM-MSCs could restore liver structure and function in CCL induced liver fibrosis rat model, ameliorating the toxicity of CCl and improving liver function tests.
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http://dx.doi.org/10.1016/j.bj.2020.04.011DOI Listing
May 2020

Reclassification of the genus : proposal of two new genera, gen. nov. to accommodate and gen. nov. to accommodate and .

Int J Syst Evol Microbiol 2020 May;70(5):3399-3405

The Institute of Life Sciences, The Hebrew University of Jerusalem, The Edmond J. Safra Campus, 9190401 Jerusalem, Israel.

The genus , currently including four species, is a member of the order , class and consists of obligately alkaliphilic and extremely halophilic members found exclusively in highly alkaline hypersaline soda lakes. The species were classified into this genus mostly based on phylogenetic analysis of the 16S rRNA gene. However, a more advanced phylogenomic reconstruction based on 122 conserved single-copy archaeal protein markers clearly indicates a polyphyletic origin of the species included into this genus, thus warranting its reclassification into three separate genera. We therefore propose to transfer (type strain N-1311) to a new genus as comb. nov. and to transfer (type strain JW/NM-HA 15) and (type strain AArc1) to a new genus as comb. nov. and comb. nov. The phylogenomic differentiation of these four species is also supported by the ANI/AAI distances and unique phenotypes. The most important physiological differences includes a previously unreported ability for cellulose and xylan utilization in , thermophily in and anaerobic sulfur respiration in . We further present an emended description of .
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http://dx.doi.org/10.1099/ijsem.0.004186DOI Listing
May 2020

The expression of zinc finger 804a (ZNF804a) and cyclin-dependent kinase 1 (CDK1) genes is related to the pathogenesis of rheumatoid arthritis.

Arch Physiol Biochem 2020 Jan 29:1-6. Epub 2020 Jan 29.

Department of Biochemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt.

ZNF804a and CDK1 genes code for proteins involved in inflammatory pathways. This study aimed to investigate the correlation of ZNF804a and CDK1 expression profiles in RA with the activity and the severity of the disease and to assess their association with inflammatory reactions in the Egyptian RA patients. ZNF804a and CDK1 expression profiles were assessed using quantitative PCR (qRT-PCR). Clinical and laboratory parameters were evaluated. ZNF804a expression was down-regulated by 0.177-fold while CDK1 expression was up-regulated to 3.29-fold in RA patients compared with healthy controls ( < .001). ZNF804a down-regulation was negatively correlated with CRP, RF, disease activity score of 28 joints (DAS) using CRP (DAS-CRP) and TNF-α. CDK1 overexpression was correlated with IFN-1 and ACPA in RA patients. ZNF804a and CDK1 genes are implicated in RA pathogenesis due to their influences on TNF-α and IFN-1 which contribute to inflammation in RA patients.
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http://dx.doi.org/10.1080/13813455.2020.1716810DOI Listing
January 2020

Thymoquinone and pentoxifylline enhance the chemotherapeutic effect of cisplatin by targeting Notch signaling pathway in mice.

Life Sci 2020 Mar 14;244:117299. Epub 2020 Jan 14.

Biochemistry Department, Faculty of Pharmacy, Suez Canal University, Egypt.

Aims: Notch signaling is highly implicated in several cancers and chemoresistance. Therefore, Notch-targeted therapies might be beneficial in enhancing chemotherapeutic effect and cancer regression. This study aimed to investigate implication of Notch in development and progression of solid Ehrlich carcinoma (SEC) and enhancement of anticancer effect of cisplatin (CIS) by addition of thymoquinone (TQ) and pentoxifylline (PTX) through modulation of Notch.

Main Methods: SEC was induced in mice as model for mammary carcinoma by s.c. injection of 1 × 10 Ehrlich cells into back of the mice. On 12 day, solid tumor was developed and mice were divided into seven groups; tumor control, early CIS (ECIS), ECIS + ETQ, ECIS + ETQ + EPTX, late CIS (LCIS), LCIS + LTQ, and LCIS + LTQ + LPTX. Early treatment was started on 12 day, whereas late treatment was begun on 19 day from tumor inoculation. At the endpoint, samples were collected for detection of Notch1, Hes1, Jagged1, β-catenin, TNF-α, IL-6, IFN-γ, IL-2, VEGF, apoptosis, CD4, and CD8.

Key Findings: Adding PTX and TQ to CIS significantly reduced Notch1, Hes1, Jagged1, β-catenin, TNF-α, IL-6, IFN-γ, and VEGF with increment in IL-2, CD4, CD8, and apoptotic cells. Moreover, early treated groups showed remarkable attenuation in tumor growth and the relevant parameters compared to their counterpart later groups.

Significance: Addition of PTX with TQ to CIS showed a synergistic chemotherapeutic action and induced better oncostatic effect mainly through Notch suppression. Consequently, shutting Notch could be of great interest in promoting chemosensetivity and cancer control.
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http://dx.doi.org/10.1016/j.lfs.2020.117299DOI Listing
March 2020

Covalent immobilization of a halophilic, alkalithermostable lipase LipR2 on Florisil nanoparticles for production of alkyl levulinates.

Authors:
Noha M Mesbah

Arch Biochem Biophys 2019 05 22;667:22-29. Epub 2019 Apr 22.

Department of Biochemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, 41522, Egypt. Electronic address:

A novel halophilic, alkalithermostable lipase LipR2 from Alkalispirillum sp. NM-ROO2 was cloned and expressed. LipR2 was covalently immobilized on Florisil functionalized with glutaraldehyde. Protein binding efficiency of functionalized Florisil was 94.7%. Immobilized LipR2 retained 97.5% of specific activity of the free enzyme. Free LipR2 has maximal activity at 52 °C, pH 9.3 and 1.9 M NaCl and is resistant to surfactants and organic solvents. Immobilization enhanced LipR2's extreme characteristics, and increased thermostability of LipR2 with the half-life at 50 °C increasing three-fold. Immobilized LipR2 was used as a biocatalyst for esterification of levulinic acid with n-butanol. Under optimal conditions, a 45.9% ester yield was obtained after 12 h. Immobilized LipR2 catalyzed production of ethyl levulinate and 1-dodecyl levulinate with 48.8% and 26.2% ester yields, respectively. When used in repetitive batch esterification, LipR2 retained 69%, 57% and 18.5% of initial activity on esterification of levulinic acid with ethanol, n-butanol and 1-dodecanol, respectively.
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http://dx.doi.org/10.1016/j.abb.2019.04.004DOI Listing
May 2019

Combined vildagliptin and memantine treatment downregulates expression of amyloid precursor protein, and total and phosphorylated tau in a rat model of combined Alzheimer's disease and type 2 diabetes.

Naunyn Schmiedebergs Arch Pharmacol 2019 06 13;392(6):685-695. Epub 2019 Feb 13.

Department of Biochemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, 41522, Egypt.

There is increasing evidence of a link between type 2 diabetes mellitus (T2DM) and cognitive decline. T2DM has been recognized as a risk factor for Alzheimer's disease (AD). The aim of this research was to investigate the biochemical and physiological effects of vildagliptin treatment alone, and in combination with memantine, in a rat model of combined T2DM and AD. The experimental study was carried out on 75 male Wistar rats weighing 180-200 g. The rats were divided into five groups (n = 15): normal group, Alzheimer diabetic control, treated with vildagliptin (10 mg/kg/day), treated with memantine (30 mg/kg/day), and treated with combination of drugs. Serum glucose, lipid profile, acetylcholinesterase (AChE), homocysteine (Hcy), and amyloid beta peptide (Aβ) were determined. Lipid peroxidation was measured in brain tissue. Expression of amyloid precursor protein (APP) in the brain was assessed by q-PCR, and expression of total and phosphorylated tau was determined by Western Blotting. Vildagliptin alone and in combination with memantine caused a decrease in blood glucose, HOMA-IR, lipid profile, Hcy, malanodialdhyde, and acetylcholinesterase, and an increase in apolipoprotein E. Expression of APP and phosphorylated tau protein was decreased with combined vildagliptin and memantine treatment. In conclusion, vildagliptin treatment, either alone or in combination with memantine, modulates AD-associated biochemical changes and downregulates amyloid precursor protein and phosphorylated tau expression in diabetic rats.
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http://dx.doi.org/10.1007/s00210-019-01616-3DOI Listing
June 2019

Differential expression of MicroRNA let-7e and 296-5p in plasma of Egyptian patients with essential hypertension.

Heliyon 2018 Nov 24;4(11):e00969. Epub 2018 Nov 24.

Department of Biochemistry, Faculty of Pharmacy, Suez Canal University, Ismailia 41522, Egypt.

Essential hypertension is a chronic medical condition affecting thousands of people worldwide. Hypertension results from interplay of genetic and environmental factors. MicroRNAs regulate gene expression and can be biomarkers for disease. MicroRNA let-7e and microRNA 296-5p have been linked to different cardiovascular diseases. This study aimed to determine association of serum miRNA let-7e and miRNA 296-5p with essential hypertension in Egyptian patients. MicroRNA let-7e and miRNA-296-5p expression was determined in sera of 25 hypertensive patients and 25 normotensive controls by quantitative real-time polymerase chain reaction. Hypertensive patients showed significantly higher expression of miRNA let-7e (3.23-fold increase, = 0.036) in comparison with normotensive controls. In hypertensive patients, miRNA let-7e expression was positively correlated with increased systolic and diastolic blood pressure. Furthermore, miRNA 296-5p expression was negatively correlated with serum total cholesterol and low-density lipoprotein. Results from this study indicate that miRNA let-7e can potentially be a biomarker for essential hypertension.
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http://dx.doi.org/10.1016/j.heliyon.2018.e00969DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6260250PMC
November 2018

Raspberry ketone preserved cholinergic activity and antioxidant defense in obesity induced Alzheimer disease in rats.

Biomed Pharmacother 2018 Nov 28;107:1166-1174. Epub 2018 Aug 28.

Ministry of Health, Zagazig, Sharkia, Egypt. Electronic address:

Obesity is a proven risk factor for neurodegenerative disease like Alzheimer's disease (AD). Accumulating evidences suggested that nutritional interventions provide potential for prevention and treatment of AD. The present study aimed to investigate the effect of dietary treatment of obese rats with natural Raspberry ketone (RK) and their relationship with neurodegeneration. Obesity was first induced in 40 male Wistar rats (140-160 g) by feeding high fat diet (HFD) for 16 weeks. Obese rats were then assigned into 4 groups (n = 10 each). (O-AD) is obese induced AD group maintained on HFD for another 6 weeks. OCR is obese group received calorie restricted diet for 6 weeks. OCRRK is obese group received calorie restricted diet and RK (44 mg/kg body weight, daily, orally) for 6 weeks and OCRD is obese group received calorie restricted diet and orlistate (10 mg/kg body weight, daily orally) for 6 weeks. Another 10 normal rats received normal diet were used as normal control group (NC). Body weight, visceral white adipose tissue weight (WAT), lipid profile, oxidative stress markers, adiponectin, cholinergic activity and amyloid extracellular plaques were examined. In addition to histological changes in brain tissues were evaluated.Raspberry ketone (RK) via its antioxidant properties attenuated oxidative damage and dyslipidemia in O-AD group. It inhibited acetylcholinesterase enzyme (AchE) and hence increased acetylcholine level (Ach) in brain tissues of O-AD rats. It is also impeded the upregulation of beta-secretase-1 (BACE-1) and the accumulation of amyloid beta (Aβ) plaques which crucially involved in AD. The combination of CR diet with RK was more effective than CR diet with orlistate (antiobese drug) in abrogating the neurodegenerative changes induced by obesity. Results from this study suggested that concomitant supplementation of RK with calorie restricted regimen effectively modulate the neurodegenerative changes induced by obesity and delay the progression of AD.
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http://dx.doi.org/10.1016/j.biopha.2018.08.034DOI Listing
November 2018

Relation of locus 1p13 rs646776 polymorphism with the risk of preeclampsia.

Hypertens Pregnancy 2018 May 25;37(2):81-86. Epub 2018 Mar 25.

a Faculty of Pharmacy, Department of Biochmistry , Suez Canal University , Ismailia , Egypt.

Objective: This study aimed to assess the relation of locus 1p13 rs646776 (T/C) polymorphism with preeclampsia in Egyptian women.

Methods: The study included 100 healthy pregnant female subjects and 100 preeclampsia patients. The genotypes of the polymorphisms were assessed. Endothelin-1 level was determined in plasma.

Results: The major T allele of the 1p13.3 genomic region rs646776 polymorphism had a higher frequency in preeclampsia patients. Carriers of C allele had significantly lower endothelin-1 levels, lower systolic and diastolic blood pressure, decreased proteinuria, and increased HDL-C in the patients.

Conclusion: The rare C allele of rs646776 polymorphism in chromosomal locus 1p13.3 is associated with decreased risk of preeclampsia.
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http://dx.doi.org/10.1080/10641955.2018.1454462DOI Listing
May 2018

The C-terminus of the ESAT6-like secretion system virulence factor EsxC mediates divalent cation-dependent homodimerization.

Toxicon 2018 May 2;146:129-137. Epub 2018 Mar 2.

Department of Microbiology and Immunology, Faculty of Pharmacy and Pharmaceutical Industries, Sinai University, El-Arish, North Sinai, Egypt. Electronic address:

The human pathogen Staphylococcus aureus encodes the ESAT6-like Secretion System (ESS). The ESS pathway secretes pathogenic substrates such as EsxA, EsxB, EsxC, EsxD and EssD that mediate staphylococcal establishment in persistent abscess lesions. The biochemical behavior of these substrates is not fully understood. EsxC is species-specific lysine-rich homodimer that lacks recognizable topogenic sequence. Studies have shown that EsxC is required for the secretion of other substrates, thereby revealing its biomedical importance. Here, EsxC self-association was investigated in the presence of several metal ion chelators. Results show that EsxC homodimerization is abolished in the presence of EDTA and EGTA, suggesting a role for calcium in mediating EsxC self-association. Complementation experiments confirm that EsxC homodimerization is calcium-dependent. N- and C-terminal truncations of EsxC were constructed, followed by bacterial two-hybrid screening. Results show that EsxC self-association is mediated by its C-terminal domain. Affinity purification of recombinant EsxC to apparent homogeneity, followed by chemical crosslinking and SDS-PAGE led to the detection of the monomeric and dimeric forms of the protein. In contrast and when a purified EsxC variant lacking the C-terminus was subjected to similar conditions, only the monomeric form was observed. These in vivo and in vitro data highlight the contribution of the C-terminus of the virulence factor EsxC to self-association, and document a previously unreported role for calcium in mediating protein-protein interactions in this pathogenic secretion system.
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http://dx.doi.org/10.1016/j.toxicon.2018.02.047DOI Listing
May 2018

Natronolimnobius aegyptiacus sp. nov., an extremely halophilic alkalithermophilic archaeon isolated from the athalassohaline Wadi An Natrun, Egypt.

Int J Syst Evol Microbiol 2018 02;68(2):498-506

Department of Microbiology, University of Georgia, Athens, GA 30602, USA.

An obligately aerobic extremely halophilic alkalithermophilic archaeon, strain JW/NM-HA 15, was isolated from the sediments of Wadi An Natrun in Egypt. Phylogenetic analysis based on 16S rRNA and rpoB' gene sequences indicated that it belongs to the family Natrialbaceae of the order Natrialbales. The closest relatives were Natronolimnobius baerhuensis IHC-005 and Natronolimnobius innermongolicus N-1311 (95.3 and 94.5 % 16S rRNA gene sequence similarity, respectively). Genome relatedness between strain JW/NM-HA 15 and its neighbours was evaluated using average nucleotide identity, digital DNA-DNA hybridization and average amino acid identity with the values of 75.7-85.0, 18.1-20.0, and 70.2-71.0%, respectively. Cells were obligately aerobic, rod-shaped, non-motile, Gram-stain-negative and chemo-organotrophic. The strain grew in the presence of 2.57 M to saturating Na (optimum 3.25-4.60 M Na), at pH 7.5-10.5 (optimum pH 9.0-9.5), and at 30-56 °C (optimum 52 °C). The major polar lipids consisted of phosphatidylglycerol, methylated phosphatidylglycerolphosphate and two phospholipids. The complete genome size of strain JW/NM-HA 15 is approximately 3.93 Mb, with a DNA G+C content of 64.1 mol%. On the basis of phylogenetic features, genomic relatedness, phenotypic and chemotaxonomic data, strain JW/NM-HA 15 was thus considered to represent a novel species within the genus Natronolimnobius, for which the name Natronolimnobius aegyptiacus sp. nov. is proposed. The type strain is JW/NM-HA 15 (=ATCC BAA-2088 =DSM 23470).
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http://dx.doi.org/10.1099/ijsem.0.002524DOI Listing
February 2018

Biochemical characterization of halophilic, alkalithermophilic amylopullulanase PulD7 and truncated amylopullulanases PulD7ΔN and PulD7ΔC.

Int J Biol Macromol 2018 May 16;111:632-638. Epub 2018 Jan 16.

Department of Microbiology, University of Georgia, Athens, GA 30602, USA.

A pullulanase, PulD7, was identified in the genome of the halophilic, alkalithermophilic isolate Alkalilimnicola sp. NM-DCM-1. PulD7 is 701 amino acids large with a carbohydrate binding module (CBM) 48 at the N-terminal. The full length PulD7 and N- and C-terminal truncated versions were cloned, heterologously expressed and functionally characterized. PulD7 displayed maximal activity at 55 °C, pH 9.5 and 2 M NaCl. PulD7 had good thermal stability, with a half-life of 693 min at 50 °C. PulD7 is an amylopullulanase, hydrolyzing both α‑1,4‑ and α‑1,6‑glycosidic bonds in soluble starch and pullulan, respectively. PulD7 was resistant to chemical reagents, including organic solvents (dimethyl sulfoxide, methanol, benzene, 20% v/v), reducing agents (β-mercaptoethanol, 5% v/v), surfactants (SDS and Tween 20, 5% v/v), the divalent chelator ethylene diamine tetra acetic acid (5 mM), and the chemical denaturant urea (8 M). PulD7 was not calcium-dependent. PulD7 was able to bind raw starch granules, reaching 52% binding in 3 h. The N-and C-terminal truncated forms of PulD7 had similar biochemical characteristics. PulD7ΔC had higher specific activity and halotolerance. The N-terminally truncated PulD7ΔN hydrolyzed amylose only, indicating that CBM48 is essential for binding branched substrates. PulD7 has unique characteristics giving it strong potential for application in biotechnological industries.
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http://dx.doi.org/10.1016/j.ijbiomac.2018.01.069DOI Listing
May 2018

Association of interleukin-1A insertion/deletion gene polymorphism and possible high risk factors with non-alcoholic fatty liver disease in Egyptian patients.

Arch Physiol Biochem 2017 Dec 18;123(5):330-333. Epub 2017 Jun 18.

a Department of Biochemistry, Faculty of Pharmacy , Suez Canal University , Ismailia , Egypt.

Context: Interleukin-1A (IL-1A) is a cytokine involved in inflammatory process. IL-1A (rs3783553) gene polymorphism is comprised in the regulation of IL-1A expression.

Objective: This study aims to evaluate association of IL-1A (I/D) gene polymorphism with NAFLD and its component traits among Egyptian populations.

Methods: The study included 75 healthy subjects and 75 patients with NAFLD. Different genotypes of IL-1A (I/D) gene polymorphism were determined by PCR-PAGE technique, serum IL-1A level and other biochemical parameters were measured.

Results: The major D allele was significantly associated with NAFLD patients (p = .002). DD genotype showed a significant increase in BMI and decrease in HDL-C. Also serum IL-1A was significantly correlated with the DD genotype. Serum IL-1A showed a significant positive correlation with BMI, triglycerides, total cholesterol, LDL-C, VLDL-C and FBG, and a significant negative correlation with HDL-C.

Conclusions: Major D allele of IL-1A (I/D) gene polymorphism is associated with NAFLD in the Egyptian population.
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http://dx.doi.org/10.1080/13813455.2017.1339717DOI Listing
December 2017

Biochemical characterization of a halophilic, alkalithermophilic protease from Alkalibacillus sp. NM-Da2.

Extremophiles 2016 Nov 18;20(6):885-894. Epub 2016 Oct 18.

Department of Biochemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, 41522, Egypt.

An extracellular, halophilic, alkalithermophilic serine protease from the halo-alkaliphilic Alkalibacillus sp. NM-Da2 was purified to homogeneity by ethanol precipitation and anion-exchange chromatography. The purified protease was a monomeric enzyme with an approximate molecular mass of 35 kDa and exhibited maximal activity at 2.7 M NaCl, pH 9 and 56 °C. The protease showed great temperature stability, retaining greater than 80 % of initial activity after 2 h incubation at 55 °C. The protease was also extremely pH tolerant, retaining 80 % of initial activity at pH 10.5 after 30 min incubation. Protease hydrolyzed complex substrates, displaying activity on yeast extract, tryptone, casein, gelatin and peptone. Protease activity was inhibited at casein concentrations greater than 1.2 mg/mL. The enzyme was stable and active in 40 % (v/v) solutions of isopropanol, ethanol and benzene and was stable in the presence of the polysorbate surfactant Tween 80. Activity was stimulated with the oxidizing agent hydrogen peroxide. Inhibition with phenyl methylsulfonylfluoride indicates it is a serine protease. Synthetic saline wastewater treated with the protease showed 50 % protein removal after 5 h. Being halophilic, alkaliphilic and thermophilic, in addition to being resistant to organic solvents, this protease has potential for various applications in biotechnological and pharmaceutical industries.
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http://dx.doi.org/10.1007/s00792-016-0879-xDOI Listing
November 2016

Intestinal fatty acid binding protein Ala54Thr polymorphism is associated with peripheral atherosclerosis combined with type 2 diabetes mellitus.

J Diabetes 2017 Sep 9;9(9):821-826. Epub 2016 Dec 9.

Department of Biochemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt.

Background: Intestinal fatty acid-binding protein 2 (FABP2) is expressed in enterocytes and binds saturated and unsaturated long-chain fatty acids. The FABP2 Ala54Thr polymorphism has been reported to effect lipid metabolism. The aim of the present study was to assess the relationship between this polymorphism and peripheral atherosclerosis combined with type 2 diabetes mellitus (T2DM) in an Egyptian population.

Methods: The study was performed on 100 T2DM patients with peripheral atherosclerosis and 100 control subjects. The Ala54Thr polymorphism was analyzed by polymerase chain reaction-restriction fragment length polymorphism, whereas serum FABP2 levels were determined using ELISA. Fasting blood glucose, fasting serum insulin concentrations, HbA1c, lipid profile, body mass index (BMI) and systolic and diastolic blood pressure (SBP and DBP, respectively) were determined.

Results: There was a higher frequency of the Thr54 allele among the patient group (P = 0.002). In Ala54/Thr54 heterozygotes and carriers of the rare Thr54/Thr54 genotype, there were significant increases in BMI and FABP2. Those with the Thr54/Thr54 genotype had significantly decreased high-density lipoprotein cholesterol (HDL-C) concentrations; in addition, those with the Thr54/Thr54 genotype had significantly higher SBP and DBP than subjects with the Ala54/Ala54 and Ala54/Thr54 genotypes. There was a positive correlation between FABP2 levels and BMI, SBP and DBP, and a negative correlation with HDL-C.

Conclusions: The Thr54 allele of the FABP2 Ala54Thr polymorphism was associated with an increased incidence of peripheral atherosclerosis combined with T2DM in the population studied.
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http://dx.doi.org/10.1111/1753-0407.12496DOI Listing
September 2017

Editorial: Enzymes from Extreme Environments.

Front Bioeng Biotechnol 2016 7;4:24. Epub 2016 Mar 7.

Swissaustral USA LLC , Athens, GA , USA.

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http://dx.doi.org/10.3389/fbioe.2016.00024DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4780105PMC
March 2016

Association of chemerin Rs17173608 and vaspin Rs2236242 gene polymorphisms with metabolic syndrome in Egyptian women.

Endocr Res 2016 15;41(1):43-8. Epub 2015 Oct 15.

a Department of Biochemistry, Faculty of Pharmacy, Suez Canal University , Ismailia , Egypt and.

Aim: The metabolic syndrome is a complex of interrelated risk factors for cardiovascular disease and diabetes. The adipokines, chemerin and vaspin, are known to have metabolic regulatory roles. This study aimed to assess the relation of chemerin rs17173608 and vaspin rs2236242 polymorphisms with metabolic syndrome and its related phenotypes in Egyptian women.

Subjects And Methods: The study included 100 healthy female subjects and 100 metabolic syndrome patients. The component traits of metabolic syndrome were determined and the genotypes of the polymorphisms were assessed using the tetra amplification refractory mutation system polymerase chain reaction procedure.

Results: The minor G allele of the chemerin rs17173608 polymorphism had a significantly higher frequency in metabolic syndrome patients (p = 0.0001). The component traits of metabolic syndrome were significantly increased in the carriers of the GG and TG genotypes. In contrast, the rare A allele of vaspin rs2236242 polymorphism was significantly higher in the control subjects (p = 0.005). The carriers of the TA and AA genotypes showed significant relation with lower values of the phenotypes of metabolic syndrome.

Conclusion: Metabolic syndrome in Egyptian females is associated with the minor allele of chemerin rs17173608 polymorphism, whereas the minor allele of vaspin rs2236242 polymorphism plays a protective role against metabolic syndrome.
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http://dx.doi.org/10.3109/07435800.2015.1066802DOI Listing
December 2016

Effects of short RNA structural analogues against hepatitis C virus genotypes 2, 3 and 4 in replicon cells.

J Dig Dis 2015 Aug;16(8):449-55

Department of Medicine, Division of Gastroenterology-Hepatology, UCONN HEALTH, USA.

Objective: To determine whether computer-predicted short RNA structural analogues could inhibit hepatitis C virus (HCV) genotype 2a, 3a and 4a replication in cultured cells.

Methods: Short RNA sequences, X12, X12a and X12b, designed to be identical in secondary structure to the X region in the 3'-untranslated region (3'-UTR) of the HCV 1b genome, as well as shorter stem-loop components of X region, were inserted into a plasmid and transfected into separate Huh7.5 human hepatoma cells stably transfected with subgenomic replicons for genotypes 2a, 3a and 4a. All replicons included a firefly luciferase reporter gene. After 48 h of plasmid transfection, the inhibition of HCV replication was determined by HCV RNA isolation and quantification by real-time polymerase chain reaction and luciferase assays.

Results: All the secondary structural analogues to genotype 1b X region cross-inhibited genotype 2a, 3a and 4a replicons. The maximum inhibition by genotype 1b X region structural analogues was obtained against genotype 2a cells in which X12, X12a and X12b inhibited replication by 30%, 63% and 72%, respectively (P < 0.05 for all), compared to an unrelated hepatitis B viral analogue.

Conclusions: Despite substantial sequence dissimilarity, HCV RNA genotype 1b X region analogues cross-inhibited the replication of HCV genotypes 2a, 3a and 4a. Particular conformations and not the sequence of the stem-loops of the X region are involved in HCV replication.
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http://dx.doi.org/10.1111/1751-2980.12250DOI Listing
August 2015

Relation of myeloperoxidase-463G/A polymorphism with metabolic syndrome and its component traits in Egyptian women.

Arch Physiol Biochem 2015 Feb 8;121(1):13-8. Epub 2014 Dec 8.

Department of Biochemistry, Faculty of Pharmacy, Suez Canal University , Ismailia , Egypt and.

Context: Myeloperoxidase is a heme protein secreted by activated macrophages and generates intermediates that oxidize lipoproteins. Myeloperoxidase-463G/A is a functional polymorphism involved in regulation of myeloperoxidase expression.

Objective: The aim of this study is to assess the relation of myeloperoxidase-463G/A polymorphism with metabolic syndrome and its component traits in Egyptian women from the Suez Canal area.

Methods: The study includes 100 healthy female subjects and 100 metabolic syndrome patients. The component traits of metabolic syndrome are determined and the genotypes of the polymorphisms assessed using the PCR-RFLP technique.

Results: There was no significant difference in the allele frequencies between the metabolic syndrome and control groups. However, the GA and AA genotypes were associated with lower total cholesterol, LDL-C, systolic and diastolic blood pressure in the patients.

Conclusion: Myeloperoxidase-463G/A polymorphism is not associated with the incidence of metabolic syndrome.
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http://dx.doi.org/10.3109/13813455.2014.988631DOI Listing
February 2015

Halophilic alkali- and thermostable amylase from a novel polyextremophilic Amphibacillus sp. NM-Ra2.

Int J Biol Macromol 2014 Sep 5;70:222-9. Epub 2014 Jul 5.

Department of Microbiology, University of Georgia, Athens, GA 30602, USA. Electronic address:

Extracellular gluco-amylo-pullulanase from Amphibacillus sp. NM-Ra2 was purified to homogeneity by ethanol precipitation, anion exchange chromatography and gel filtration chromatography. Molecular mass of the enzyme was 50kDa (SDS-PAGE). The enzyme showed maximal activity at 1.9 M NaCl, pH50°C 8.0 and 54°C and was active from 0 to 4.3 M NaCl and 37 to 65°C. The enzyme was inhibited by EDTA and was stable and active in the presence of PMSF, DTT, H2O2, Triton-X-100, Tween 20 and Tween 80. Ca2+ is inessential for activity. The amylase was stimulated with K+ and inhibited with Cu2+ and Mg2+. Hg2+, Zn2+ and Fe2+ had no effect on activity. Amylase was stable and active in the presence of ethanol, methanol and benzene (25%, v/v). The enzyme hydrolyzed linear and branched polysaccharides including pullulan, glycogen and amylopectin, and hydrolyzed raw wheat starch and raw corn starch (14.6% and 13.5% over 2 h). Amylase activity was inhibited by soluble starch concentrations greater than 0.3%. The major products of soluble starch hydrolysis were maltose and maltotriose. The amylase, being halophilic and alkali-thermostable, in addition to being resistant to surfactants, oxidizing agents and organic solvents, can find applications in the starch processing, pharmaceutical, food and paper/pulp industries.
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http://dx.doi.org/10.1016/j.ijbiomac.2014.06.053DOI Listing
September 2014

Association of estrogen receptor alpha gene polymorphisms with metabolic syndrome in Egyptian women.

Metabolism 2013 Oct 25;62(10):1437-42. Epub 2013 Jun 25.

Department of Medical Biochemistry, Faculty of Medicine, Suez Canal University, 41522 Ismailia, Egypt.

Objective: Metabolic syndrome is a risk factor for coronary heart diseases as well as diabetes, fatty liver and several cancers. The prevalence of metabolic syndrome in women appears to be increasing, particularly in women of childbearing age. In the present study, we assessed the association of estrogen receptor-alpha gene polymorphisms (XbaI and PvuII) with metabolic syndrome and its related phenotypes.

Materials/methods: One hundred and fifty Egyptian female patients with metabolic syndrome (mean age 35.52±6.86) were compared with one hundred and fifty age matched healthy Egyptian women (controls). The component traits of metabolic syndrome were determined, and the XbaI and PvuII genotypes were assessed with the PCR-RFLP method.

Results: Our data indicated a significant difference in the allele frequencies of XbaI, but not PvuII, between the metabolic syndrome and control groups (P=0.0003 and P=0.164). Carriers of the minor alleles of XbaI and PvuII gene polymorphisms, in either the homozygous or heterozygous form, were associated with high diastolic blood pressure, high total cholesterol and LDL-c levels, increased HOMA-IR values and decreased QUICKI values compared to carriers of the major allele. However, only the minor G allele of XbaI was associated with measures of adiposity, specifically, BMI and waist circumference.

Conclusions: The XbaI polymorphism of the estrogen receptor alpha gene is associated with metabolic syndrome. On the other hand, PvuII gene polymorphism is not associated with the occurrence of the disease in this sample of Egyptian women.
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http://dx.doi.org/10.1016/j.metabol.2013.05.014DOI Listing
October 2013

Relaxin-3 is associated with metabolic syndrome and its component traits in women.

Clin Biochem 2013 Jan 24;46(1-2):45-8. Epub 2012 Sep 24.

Department of Medical Biochemistry, Faculty of Medicine, Suez Canal University, 41522 Ismailia, Egypt.

Objectives: Relaxin-3 was found to play a role in appetite regulation, increasing food intake and body weight. The current study aimed to investigate the relation of relaxin-3 to metabolic syndrome and its component traits in women.

Design And Methods: The study was conducted on 300 female subjects, 150 healthy control and 150 metabolic syndrome patients. The component traits of metabolic syndrome were determined for all participants.

Results: Serum relaxin-3 level was significantly higher in the metabolic syndrome patients than in the healthy control group. It was also significantly correlated with all the component traits of metabolic syndrome.

Conclusion: The results suggest that metabolic syndrome is associated with increased serum relaxin-3 levels in women. Relaxin-3 might be considered as a potential biomarker of metabolic syndrome.
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http://dx.doi.org/10.1016/j.clinbiochem.2012.09.018DOI Listing
January 2013

Life under multiple extreme conditions: diversity and physiology of the halophilic alkalithermophiles.

Appl Environ Microbiol 2012 Jun 6;78(12):4074-82. Epub 2012 Apr 6.

Department of Biochemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt.

Around the world, there are numerous alkaline, hypersaline environments that are heated either geothermally or through intense solar radiation. It was once thought that such harsh environments were inhospitable and incapable of supporting a variety of life. However, numerous culture-dependent and -independent studies revealed the presence of an extensive diversity of aerobic and anaerobic bacteria and archaea that survive and grow under these multiple harsh conditions. This diversity includes the halophilic alkalithermophiles, a novel group of polyextremophiles that require for growth and proliferation the multiple extremes of high salinity, alkaline pH, and elevated temperature. Life under these conditions undoubtedly involves the development of unique physiological characteristics, phenotypic properties, and adaptive mechanisms that enable control of membrane permeability, control of intracellular osmotic balance, and stability of the cell wall, intracellular proteins, and other cellular constituents. This minireview highlights the ecology and growth characteristics of the extremely halophilic alkalithermophiles that have been isolated thus far. Biochemical, metabolic, and physiological properties of the extremely halophilic alkalithermophiles are described, and their roles in resistance to the combined stressors of high salinity, alkaline pH, and high temperature are discussed. The isolation of halophilic alkalithermophiles broadens the physicochemical boundaries for life and extends the boundaries for the combinations of the maximum salinity, pH, and temperature that can support microbial growth.
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http://dx.doi.org/10.1128/AEM.00050-12DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3370554PMC
June 2012

Molecular and protein characterization of two species of the latrunculin-producing sponge Negombata from the Red Sea.

J Pharm Biomed Anal 2011 Dec 30;56(5):911-5. Epub 2011 Jul 30.

Department of Biochemistry, Faculty of Pharmacy, Suez Canal University, Ismailia 41522, Egypt.

Red Sea sponges offer a potential for production of novel drugs and prototypes. The genus Negombata is a type of sponges abundant in the Red Sea. This sponge produces latrunculins that have well documented antitumor activity in addition to antimicrobial and antiviral effects. The identification of Negombata species is based on morphology and microscopical examination of megascleres of spicules. However, these criteria have proven to be unreliable for identification. Therefore, this study was established to test the accuracy of the spicules based taxonomy against molecular and protein profiles for the two species of Negombata: N. magnifica and N. corticata. About 700 bp of cytochrome c oxidase I gene was sequenced from the tissues of the two Negombata species. Additionally total proteins were extracted from Negombata samples collected from different locations during different seasons and separated by denaturing polyacrylamide gel electrophoresis. Characteristic different protein profiles were obtained for both species. The data obtained from cytochrome c oxidase I gene sequencing and protein profiles can reliably differentiate between different species of Negombata in the Red Sea.
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http://dx.doi.org/10.1016/j.jpba.2011.07.037DOI Listing
December 2011

Complete genome sequence of the anaerobic, halophilic alkalithermophile Natranaerobius thermophilus JW/NM-WN-LF.

J Bacteriol 2011 Aug 3;193(15):4023-4. Epub 2011 Jun 3.

Department of Microbiology, University of Georgia, Athens, GA 30602, USA.

The genome of the anaerobic halophilic alkalithermophile Natranaerobius thermophilus consists of one 3,165,557-bp chromosome and two plasmids (17,207 bp and 8,689 bp). The present study is the first to report the completely sequenced genome of an anaerobic polyextremophile and genes associated with roles in regulation of intracellular osmotic pressure, pH homeostasis, and growth at elevated temperatures.
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http://dx.doi.org/10.1128/JB.05157-11DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3147539PMC
August 2011

The Na(+)-translocating F₁F₀-ATPase from the halophilic, alkalithermophile Natranaerobius thermophilus.

Biochim Biophys Acta 2011 Sep 10;1807(9):1133-42. Epub 2011 May 10.

Department of Microbiology, University of Georgia, Athens, GA 30602, USA.

Natranaerobius thermophilus is an unusual anaerobic extremophile, it is halophilic and alkalithermophilic; growing optimally at 3.3-3.9M Na(+), pH(50°C) 9.5 and 53°C. The ATPase of N. thermophilus was characterized at the biochemical level to ascertain its role in life under hypersaline, alkaline, thermal conditions. The partially purified enzyme (10-fold purification) displayed the typical subunit pattern for F-type ATPases, with a 5-subunit F(1) portion and 3-subunit-F(O) portion. ATP hydrolysis by the purified ATPase was stimulated almost 4-fold by low concentrations of Na(+) (5mM); hydrolysis activity was inhibited by higher Na(+) concentrations. Partially purified ATPase was alkaliphilic and thermophilic, showing maximal hydrolysis at 47°C and the alkaline pH(50°C) of 9.3. ATP hydrolysis was sensitive to the F-type ATPase inhibitor N,N'-dicylohexylcarbodiimide and exhibited inhibition by both free Mg(2+) and free ATP. ATP synthesis by inverted membrane vesicles proceeded slowly and was driven by a Na(+)-ion gradient that was sensitive to the Na(+)-ionophore monensin. Analysis of the atp operon showed the presence of the Na(+)-binding motif in the c subunit (Q(33), E(66), T(67), T(68), Y(71)), and a complete, untruncated ε subunit; suggesting that ATP hydrolysis by the enzyme is regulated. Based on these properties, the F(1)F(O)-ATPase of N. thermophilus is a Na(+)-translocating ATPase used primarily for expelling cytoplasmic Na(+) that accumulates inside cells of N. thermophilus during alkaline stress. In support of this theory are the presence of the c subunit Na(+)-binding motif and the low rates of ATP synthesis observed. The complete ε subunit is hypothesized to control excessive ATP hydrolysis and preserve intracellular Na(+) needed by electrogenic cation/proton antiporters crucial for cytoplasmic acidification in the obligately alkaliphilic N. thermophilus.
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http://dx.doi.org/10.1016/j.bbabio.2011.05.001DOI Listing
September 2011
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