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OBJECTIVE: The current study evaluates the effect of metformin (MET) and /or alpha lipoic acid (ALA) on hypothyroidism and its adverse effects on the cardiac, renal, and, hepatic functions in rats. MATERIALS AND METHODS: Rats were divided into five groups: control, rat model of hypothyroidism induced by propylthiouracil (PTU), rat model of hypothyroidism treated with MET, rat model of hypothyroidism treated with ALA, and rat model of hypothyroidism treated with MET and ALA. At the end of the experiment, body weight gain was determined and the blood samples were collected from orbital plexus to measure the serum levels of thyroxine (T4), triiodothyronine (T3) and thyroid stimulating hormone (TSH) by ELISA, glucose level, the activities of lactate dehydrogenase (LDH), creatine kinase MB (CK-MB), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP), and the levels of urea and creatinine spectrophotometrically. RESULTS: Rat model of hypothyroidism revealed a significant decrease in T4 (p

An aqueous solution of magnetite (Fe3O4) nanoparticles was synthesized using the method of co-precipitation. The nanoparticles were activated with epichlorohydrin for covalently immobilizing the catalase enzyme. The immobilization conditions were optimized as 0.07 mg/ml catalase for 1 h contact time. The properties of free and immobilized catalase were also investigated. The immobilized enzyme showed enhanced activity at alkaline pH and retained about 90% of its relative activity between pH (6–8) and resisted the high temperature and retained 90% of its relative activity at 50 °C. Kinetic parameters of free and immobilized catalase were investigated. While the Vmax value of the immobilized enzyme was reduced 2.4 fold compared to the free enzyme, the KM value of the immobilized catalase was higher by 2.2 fold than the free enzyme. The formulated matrix enhanced the velocity of the immobilized catalase more than the free one and was able to be used for about 18 cycles with retention of 80% of its activity. The immobilized catalase on epoxy functionalized iron oxide nanoparticles is promising as a nano-bio-catalyst carrying out in many industries and different fields.

Objective The purpose of the current study was to investigate the possible anti-tumor effect of miR-27a inhibitor in combination with Sorafenib (SOR) on cell proliferation and apoptosis of hepatocellular carcinoma cell lines. Methods Transient transfection by oligo-miR27a inhibitor (miR-27ai) was used in this study for targeting the oncogenic miR-27a in HepG2 and Huh7 cells followed by SOR treatment. Cell viability was measured using SRB assay. The cell cycle and apoptosis were assessed by flow cytometry assay. Moreover, the level of oncogenic miR-27a was evaluated in 19 tissues of primary HCC patients as well as cell lines using qRT-PCR assay. Finally, caspase-3 activity was determined using ELISA assay. Results Significant up-regulation of miR-27a expression was reported in HCC patients confirming its oncogenic role. Treatment of cells with SOR following transfection with miR-27ai declined cell viability significantly compared with either control or single agent treatment (p≤0.05). Highly significant decreasing in the number of cell in S-phase associated with increasing in G0-phase was also observed. Furthermore, apoptotic rate was highly significantly increased for transfected/SOR treated cells (p≤0.01). Finally, combination treatment demonstrated a significant elevation of caspase-3 activity level in both cell lines examined. Conclusion The present data demonstrated targeting miR-27a enhances the anti-tumor effect of SOR in HCC cell lines considering as one of the promising therapeutic targets for advanced HCC management.

Objective: Cancer treatment using a targeted inducer of apoptosis like tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) faced the obstacle of resistance, thus providing a plus drug like Thymoquinone (TQ) could be of great interest to tackle breast cancer cells. The aim of the present work is to examine the genetic modulation impacts of the TRAIL receptors and apoptotic markers upon the combinatorial remedy of TRAIL plus TQ on human breast cancer cell lines. Methods: To achieve this rationale, the protein content-based cytotoxicity using SRB assay, as well as the genetic expressions of the TRAIL receptors (DR4 and DR5) and apoptotic markers (Bcl-2, Cas-8, and FADD) using real time qRT-PCR technique were preceded against breast cancer MCF-7 and MDA-MB-231 cancerous cell lines. Results: The current study showed that the combination therapy of TQ+TRAIL significantly inhibited the protein content-based proliferation of MDA-MB-231 cells more than MCF-7 cells. The synergistic effect of them significantly up-regulated the genetic expressions of DR4, DR5, Cas-8, and FADD genes and inhibited the genetic expression of the Bcl-2 gene in the proposed cell lines treated for 24 h. The induction of the apoptotic genes using the combined therapy was stimulated by the elevation of the reactive oxygen species (ROS); nitric oxide (NO) and malondialdehyde (MDA) levels. Conclusions: The synergistic influence between TQ which induced the DR5 and TRAIL, facilitating the connection between TRAIL and its receptors on the cancerous cell membrane. Hence, the proposed combination therapy induced the ROS-mediated apoptotic stimulus.

Objective: Cancer is one of the leading causes of mortality in both developed and developing nations. The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is characterized by its ability to selectively trigger apoptosis in cancer cells. TRAIL-based interventions have led to the development of recombinant human (rhTRAIL) as a promising therapy for different types of human cancer. Thymoquinone (TQ) has been shown to exert anticancer effect. The aim of the current study is to investigate the anticancer effect of the combinatorial therapy of TRAIL+TQ against human breast cancer cells. Methods: To achieve this hypothesis, cytotoxicity using MTT assay, as well as apoptosis and cell cycle using flow cytometric technique were preceded against breast cancer MCF-7 and MDA-MB-231 cancerous cell lines. Results: The current study showed that TRAIL induced cell cycle arrest and apoptosis. Moreover, it inhibited proliferation of MDA-MB-231 cells more than MCF-7 cells. Adding TQ to TRAIL increased the chemo-sensitivity of MDA-MB-231, while overcame the MCF-7 resistance to TRAIL. Conclusion: In conclusion, there is a synergistic effect between TRAIL and TQ playing a therapeutic role in killing resistant breast cancer cells.

Filamentous fungi have been widely utilized in production of enzymes which have many industrial applications. In this study, twenty five local fungal isolates, belonging to Aspergillus sp., Trichoderma sp. and Penicillium sp., were screened under solid state fermentation conditions (SSF) for the production of α-amylase, glucoamylase and cellulase. Asperigillus oryzae F-923, cultivated on wheat bran, was the most promising isolate for production of the target enzymes under this study. Physical parameters of moisture content, pH, temperature and incubation time, optimized were 1:2(w/v), 5.5, 28°C and 72 hr, respectively. The production of enzymes was enhanced when ammonium sulfate was supplemented as a nitrogen source to wheat bran. The production of α-amylase and glucoamylase was also enhanced when 10% (w/w) soluble starch was added as a carbon source to wheat bran. However xylose supplementation at 10% (w/w) was observed to be best for cellulase production. Tap water was found to be efficient for enzymes' extraction from the fermentation medium. Three successive extractions were needed to obtain the produced enzymes from the fermented substrate. Characterization of the produced enzymes revealed that, the optimum temperature for α-amylase and glucoamylase was 60°C, while 50°C was the optimum temperature for cellulase activity. Isopropanol 1:1(v/v) was proved to be more suitable for partial purification of enzymes. Following partial purification of enzymes of glucoamylase, α-amylase and cellulase increased to 10.8, 11.8 and 11.4 folds, respectively.

Objective: The present study evaluates the neuroprotective effect of α-lipoic acid (ALA) and/or metformin (MET) on the behavioral and neurochemical changes induced by hypothyroidism. Methods: Rats were divided into control, rat model of hypothyroidism induced by propylthiouracil, and rat model of hypothyroidism treated with ALA, MET, or their combination. Results: Behaviorally, hypothyroid rats revealed impaired memory and reduced motor activity as indicated from the novel object recognition test and open-field test, respectively. Hypothyroidism induced a significant increase in lipid peroxidation (malondialdehyde [MDA]) and a significant decrease in reduced glutathione (GSH) and nitric oxide (NO) in the cortex and hippocampus. These were associated with a significant increase in tumor necrosis factor-α (TNF-α) and a significant decrease in brain-derived neurotrophic factor (BDNF). Hypothyroidism decreased significantly the levels of serotonin (5-HT), norepinephrine (NE), and dopamine (DA) and reduced the activities of acetylcholinesterase (AchE) and Na+, K+-ATPase in the cortex and hippocampus. Treatment of hypothyroid rats with ALA and/or MET showed an improvement in memory function and motor activity. Moreover, ALA and/or MET prevented the increase in MDA and TNF-α, and the decline in GSH, NO, BDNF, 5-HT, NE, and DA. It also restored AchE and Na+, K+-ATPase activities in the studied brain regions. Conclusion: ALA and/or MET has a potential neuroprotective effect against the adverse behavioral and neurochemical changes induced by hypothyroidism in rats.

Novel three-dimensional biodegradable porous nanocomposite bone scaffolds were fabricated using acacia gum and gelatin as the base polymer matrix and magnesium doped nano hydroxyapatite as cementing materials using gamma irradiation facility for crosslinking and sterilization processes. Mg-doped HAp nanoparticles were synthesized using wet chemical method. XRD studies verified the nano-scale size of the prepared HAp. In addition to Ca and P in the prepared n-HAp, the EDX analysis revealed the presence of Mg in the doped HAp samples. FTIR studies confirmed the existence of the characteristic functional groups of the scaffold constituents. The swelling behavior was found to be dependent on the quantity of embedded HAp nanoparticles. Nanocomposite scaffold porosity ranged from 26 to 39%, which increased with the inclusion of Mg ions. The developed scaffolds showed appropriate mechanical properties that enhanced by the existence of HAp nanoparticles. The incorporation of the Mg-doped HAp nanoparticles encourages the development of bone-like apatite layer. In vitro cytotoxicity assessment and blood compatibility demonstrated their biocompatibility. The developed scaffolds show promising antibacterial activity against Staphylococcus aureus and Escherichia coli. In vitro drug release study showed that the loaded Ketoprofen scaffolds were able to deliver the loaded drug sustainably.

MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression through post-transcriptional interactions with mRNA. MiRNAs have recently been considered as key regulators of various cancers including liver cancer. Sorafenib is one of the antitumor drugs for the treatment of advanced hepatocellular carcinoma. It acts as a multikinase inhibitor suppressing cell proliferation and angiogenesis. This study tries to investigate a potential microRNA-based mechanism of action of the drug by studying the effect of sorafenib on miR-23a and miR-24 levels in HCC cell lines HepG2 /Huh7 and revealing the possible drug mechanism against these oncogenic mi-RNAS in this study cell viability of cultured HepG2 /Huh7 after treatment with sorafenib were evaluated using Sulphorhodamine-B (SRB) assay, cell cycle, and apoptosis estimated by flow cytometry assay. The caspase-3 level was determined using the ELISA assay. Moreover, miR-23a and miR-24 expression levels were analyzed by qPCR. Finally, TGF-β levels and phosphorylated smad2, 3 were examined after treatment with sorafenib using ELISA and western blotting. Our data confirmed the Sorafenib inhibition of cell growth in both cell lines which was accompanied by a significant increase in cell apoptosis and cell cycle arrest. Cells treated with sorafenib showed a significant decrease in miR-23a and miR-24 levels in both cell lines. Interestingly, the change in these oncogenic miRNAs was accompanied by a significant decrease of (TGF-β1) and phosphorylated smad2, 3 protein levels. Our study suggested that inhibition of the tgf beta pathway in smad dependent manner could be the way characteristic of sorafenib to inhibit the oncogenic miR-23a and miR-24 levels in HCC.

A new matrix formulation was devised for catalase immobilization. Carrageenan-alginate beads different ratios were developed and soaked into different ratios of CaCl(2)–KCl as a hardening solution. The best formulation for loading capacity was selected, treated with polyethylene imine followed by glutaraldehyde and further studied. The best concentration of catalase for immobilization was 300U/ml and the best loading time was 6 h. The catalytic properties increased after immobilization and the immobilized catalase achieved optimum activity at a temperature range of 30–50 °C that was compared to the optimum activity of free catalase which occurred at 40 °C. Higher catalytic activity of immobilized catalase occurred at alkaline pHs than the free one which achieved optimum catalytic activity at neutral pH. A comparison between the kinetic parameters of immobilized and free catalase showed variation. The K(M) and Vmax of the immobilized catalase were 2.4 fold and six times higher than those of free catalase. The results of the study indicate that the formulated matrix can be used as a good matrix for catalase enzyme in various industrial applications.

Objectives Stunting affects 32% of children living in developing countries and has a major impact on child health and development. In this study, we aimed to assess the pro-inflammatory cytokines and some micronutrients among the healthy control and nutritionally stunted Egyptian children. Subjects and methods A total of 88 children were enrolled; 60 stunted children and 28 non-stunted of matched age and sex. Clinical, demographic characteristics, serum levels of calcium, magnesium, and zinc were measured. Plasma levels of interleukin-6 (IL-6), tumor necrosis factor alpha (TNFα) and C – reactive protein (CRP) were measured in these subjects. Results Stunted children showed a marked decrease in weight and weight-for-age Z score (WAZ) versus healthy one. Significant reduction in the serum level of Ca, Mg, and Zn was detected in stunted children compared to those of the healthy control subjects. Moreover, the mean serum levels of IL-6 (pg/ml) (1.5 Vs 1.6), TNFα (pg/ml) (1.7 Vs 1.8) and CRP (mg/l) (0.7 Vs 1) were significantly higher among the stunted group compared to control. Conclusion Nutritionally stunted Egyptian children have lowered serum levels of Zn. Ca, and Mg, in addition to elevated serum levels of proinflammatory cytokines (Il-6, TNFα, and CRP).

In this study, a fungal cellulase enzyme was applied to improve the selective separation of apatite from hematite during the reverse anionic flotation of a high-phosphorus iron ore. Bench-scale flotation tests were conducted on pure hematite and apatite minerals, their artificial mixture, and a high-phosphorus iron ore sample, using oleic acid as a collector. Flotation tests on single minerals showed that cellulase was a good depressant of hematite, but had no depressing effect on apatite at pH 10. The phosphorus content in the concentrate product of the flotation of artificially mixed pure minerals was reduced from 1.96% to 0.03%, with iron grade and recovery of 64.15% and 94.74%, respectively, using 4x10−5 mol/L oleate and 0.02% cellulase at pH 10. By applying the optimum separation conditions to the flotation of a natural high-phosphorus iron ore, the phosphorus content in the concentrate was reduced from 0.84% to 0.19% at an iron recovery of 67.85%. The mechanism of the cellulase depressing effect was investigated through Fourier transform infrared spectroscopy (FTIR), and X-ray photoelectron spectroscopy (XPS) analyses. The results showed that cellulase enzyme was adsorbed on the surfaces of both minerals; however, the depressing effect was significant in case of hematite but not apatite. This might be attributed to the coverage of apatite surface with a close-packed layer of oleate molecules, which made the surface of apatite hydrophobic enough to counter the polarity induced upon cellulase adsorption.

Childhood nutritional stunting, an indicator of chronic malnutrition, has been suggested as one factor that can contribute to high incidences of obesity, and impaired lipid and glucose metabolism in developing countries. This study aimed to determine health risks of developing hyperlipidemia, obesity, and diabetes as non-communicable diseases (NCDs) among nutritionally stunted Egyptian children. A total of 90 children were enrolled; 60 nutritional stunted children and 30 non- stunted of matched age and sex. Clinical, demographic characteristics were determined for the studied children. Lipid profile levels, fasting plasma levels of glucose, and insulin were measured in these subjects. Pancreatic beta cells’ function (HOMA-β) and insulin resistance (HOMA-IR) were calculated by homeostasis model assessment (HOMA). Stunted children exhibited significantly lower values for weight, weight-for-age Z score (WAZ), height, and height-for-age Z score (HAZ) as compared to normal ones. Significant elevation in total cholesterol (TC), and low density lipoprotein cholesterol (LDL-c), significant decrease in high density lipoprotein cholesterol (HDL-c) and HDL/LDL ratio were detected compared to that of the healthy control subjects. There were no significant differences in the levels of fasting glucose, fasting insulin, homeostasis model assessment of beta-cell function (HOMA-β), and homeostasis model assessment of insulin resistance (HOMA-IR) between the two compared groups. Conclusion: Alterations in lipid pattern among nutritionally stunted Egyptian children may predict the future of NCDs later in life.

The interaction of oleic acid, as an anionic detergent, with cellulase has been investigated using different techniques (surface tension, turbidity and FTIR). The surface tension measurements show that the CMC of the oleic acid-cellulase mixture is lower than that of the individual components. The interaction in this case is defined as attractive and the mixture exhibits synergism. Also, it is found that the area per molecule for the oleic acid/cellulase mixed isotherm is lower than area of the pure oleic acid. This decrease in the area/molecule is due to the arrangement of cellulase molecules in-between the oleic acid chains. Turbidity measurements indicate the formation of two complexes. Complex 1 is obtained at low oleic concentrations (< 10−4 M) with compact size giving a high turbidity. Complex 2 is formed at higher oleic concentrations (> 10−4 M) where the formed aggregates were precipitated and the turbidity was decreased. The changes in the FTIR spectrum features of cellulase after oleic acid addition indicate an interaction between cellulase and oleic acid in their mixture.

Development of controlled drug release carriers with gastroretentive ability is one of the most feasible approaches to improve the bioavailability and efficiency of medical treatment of Helicobacter pylori-induced peptic ulcers. Consequently, modification of alginate with N, N-dimethylamino ethyl methacrylate was occurred via graft copolymerization technique using γ-irradiation. (Alg-g-DMAEMA) copolymers were used to prepare Amoxicillin-Trihydrate floated hollow beads using calcium chloride as crosslinker and calcium carbonate as a gas-forming agent to induce floatability. Swelling behavior, floatability, and gastric retention time were investigated in simulated gastric fluid. Morphological features, drug encapsulation efficiency, and in vitro (AT) release were evaluated. Beads show internal hollow construction with bulk density ranged from 0.393 to 1.533 (g/cm3) and enhanced gastric residence time extended up to 24 h. Drug entrapment efficiency ranged between 63 to 97% with sustainable release behavior extending beyond 15 h depends on beads p (DMAEMA) content. The newly developed pH-sensitive floated (AT) loaded (Alg-g-DMAEMA) beads are promising for increasing gastric residence time with sustained (AT) release which improves its bioavailability for the effective treatment of H. pylori. The in-vitro study showed that (AT) loaded beads eradicate more than 95% of H. pylori after 10 h.

The aim of this study was to investigate the radioprotective effect of salicin against irradiation effect on spleen tissue in male rats. Lipid peroxidation product (MDA) level was measured as thiobarbituric acid reactive substance. Ultrastructural examination was carried out in spleen tissue by scanning electron microscope (SEM). The polyacrylamide gel electrophoresis for native protein, lipoprotein and zymogram were carried out in spleen homogenate. As expected, salicin resisted the irradiation effect and declined the MDA level in spleen homogenate of all treated groups. The alterations which were occurred as a result of irradiation in the spleen tissue could not be detected microscopically but they were detected electrophoertically at levels of protein and isozymes. Salicin prevented the qualitative mutagenic effect of irradiation on the electrophoretic protein pattern in the irradiated salicin simultaneous treated group (SI = 0.73). It showed the highest protective effect against qualitative mutagenic irradiation effect in catalase pattern in irradiated salicin pre-treated group (SI = 0.80). It could not prevent the abnormalities occurred qualitatively and quantitatively as a result of irradiation in peroxidase pattern in all irradiated salicin treated groups. While the esterase pattern showed the same electrophoretic pattern in the all irradiated salicin treated groups. The results suggested the radioprotective ability of salicin against gamma irradiation effect on various ultrastructural and electrophoretic patterns in spleen tissue of male rats.

The aim of this study was to investigate the radioprotective effect of salicin against irradiation effect on brain tissue of male rats. Lipid peroxidation level was measured as thiobarbituric acid reactive substance in brain tissue. The polyacrylamide gel electrophoresis for native protein, lipoprotein and zymogram were carried out in brain homogenate. As expected, salicin resisted the irradiation effect and declined the MDA level in brain homogenate of all treated groups (especially in the irradiated salicin post-treated group). Salicin minimized the qualitative mutagenic effect of irradiation on the electrophoretic protein pattern in all irradiated salicin treated groups and it showed the highest antagonistic effect against irradiation in irradiated salicin post-treated group (SI = 0.57). It could not prevent the abnormalities occurred qualitatively and quantitatively as a result of irradiation in lipoprotein pattern in all irradiated salicin treated groups. In the electrophoretic esterase pattern, salicin prevented the qualitative effect of irradiation in irradiated salicin post-treated group (SI = 1.00). Salicin minimized the qualitative irradiation effect on the catalase pattern in the irradiated salicin pre-treated group (SI = 0.73). While in the peroxidase pattern, salicin adminstration resisted the irradiation effect in the irrradiated post-treated groups (SI = 0.67). The results suggested the radioprotective ability of salicin against gamma irradiation effect on various electrophoretic patterns in brain tissue of male rats.

Drugs used for treatment of cancerous diseases by mean of chemotherapy are often limited due to their severe undesirable side effects in multiple organs. Cyclophosphamide (CYP) belongs to class of the cytotoxic bifunctional alkylating agents. In the current study, it was revealed that CYP caused significant (P˂0.05) elevation in levels of different biochemical measurements. Carob extract exhibited beneficial effect through lowering of all elevated parameters. Furthermore, CYP caused decline in total antioxidant capacity (TAC) level in association with elevation of lipid peroxidation (LPO) level and significantly (P˂0.05) in liver tissue. Carob extract restored TAC and lowered LPO level. CYP caused several histopathological alterations in the hepatocytes and carob extract minimized severity of these alterations. The similarity index percent (SI %) in the native electrophoretic protein, lipoprotein and calcium moiety of protein patterns was represented by lowest values (87.50, 66.67 and 70.59, respectively) with the CYP-treated group. The SI % values increased in all carob treated groups. In all electrophoretic isoenzymes, the SI % value was represented by the lowest value (60.00, 66.67, 66.67 and 53.33 respectively with catalase (CAT), peroxidase (GPx), α- and β-esterase (EST) patterns) in CYP-treated group. While in all carob treated groups, the SI % value reached the highest value (100.00). Furthermore, CYP induced cleavage of the genomic DNA and the carob extract maintained the DNA integrity. The study concluded that carob showed ameliorative effect against alterations induced by CYP at biochemical, histopathological and molecular levels in liver tissue of rats.

Background and Aims Hepatitis C virus (HCV) has emerged as the major pathogen of liver disease worldwide. The aim of this study was to quantitate and evaluate the performance of HCV-NS4 antigen as an alternative approach for confirmation of viremia. Methods Detection of HCV-NS4 was assessed in 883 patients with chronic hepatitis C. Areas under the ROC curves (AUC) were used to assess and compare diagnostic accuracy of ELISA for HCV-NS4 with quantitative HCV-RNA as a gold standard. Results HCV-NS4 was identified at 27 kDa using Western blot. AUC for HCV-NS4 detection was 0.95 for all patients with different liver pathologies: 0.93 for liver fibrosis (LF), 0.95 for liver cirrhosis (LC) and 0.98 for hepatocellular carcinoma (HCC). The mean ± SD (μg/mL) of HCV-NS4 in LF was 94.2 ± 55.6; in LC was 99.3 ± 64.8 and in HCC was 124.9 ± 70.3. Conclusions HCV-NS4 antigen detection using ELISA is a reliable test in the confirmation of HCV infection.

Background and Aim: Response to interferon therapy and disease progression in hepatitis C virus (HCV) infected patients differs among individuals, suggesting a possibility of a contribution of host genetic factors. 2′-5′-oligoadenylate synthetase 1 (OAS1), an important component of the innate immune system with a proven antiviral function, may therefore have a relationship with the response to interferon therapy and clinical course of HCV disease. Our aim was to determine the frequency of single nucleotide polymorphism (SNP) at exon 7 splice acceptor site (SAS) of the OAS1 gene in relation to the interferon response and status of HCV infection. Methods: A 203 bp fragment containing exon 7 SAS was amplified in 70 HCV chronic patients and 50 healthy controls. SNP was examined using restriction fragment length polymorphism (RFLP) genotyping method. Correlations of SNP genotypes with response to interferon and clinical status of patients were statistically analyzed. Results: There was an increasing trend of response from AA to AG to GG genotypes (P = 0.007). Genotype AA was associated with non-response to interferon and higher degree of liver fibrosis (P = 0.05). Multivariate analysis showed this SNP as independent and a significant determinant of the outcome of interferon therapy (odds ratio 4.913 [95% confidence interval 1.365–8.2], P = 0.006). Conclusions: This is the first study to show a significant association between the functional SNP at exon 7 SAS of OAS1 gene and the viral response to interferon in chronic HCV patients. Patients with AA genotype were associated with progressive HCV disease and viral resistance to interferon therapy. This OAS SNP is a potential bio-marker to predict IFN response in chronic hepatitis C patients.

Plasma Vascular endothelial growth factors (p VEGFs) as well as plasma insulin like growth factors- I (pIGFs- I) has been implicated in the pathogenesis of diabetes mellitus. This study was performed to determine whether alternations of p VEGFs and pIGFs are related to diabetic nephropathy in type 2 diabetic patients.Patients & Methods:We examined the association of pVEGFs and pIGFs concentrations with fasting glucose levels, glycosylated hemoglobin (HbA1c %), urinary measured renal parameters i.e. creatinine clearance and albuminuria in 75 patients with type 2 diabetes and 25 healthy controls. Study subjects were divided into four groups using urinary albumin-to-creatinine ratio (ACR). Results:We confirmed that (i) both pVEGFs and pIGFs showed remarkable increase in all diabetic groups with worsen A/Cr ratio, as compared with controls. (ii) p VEGFs and pIGFs were increased in diabetic patients as long as glycemic control was not achieved. (iii) Vascular endothelial growth factor in plasma as well as plasma insulin like growth factors elevations were also revealed statistically. (iv) Direct positive correlation between pVEGFs and pIGFs-I with glycemic control index, albuminuria were noticed. Conclusion:The release of both p VEGFs as well as pIGFs was increased during the earlier stage of diabetic nephropathy and were significantly correlated with urinary albumin excretion. This suggested that pVEGFs could be used as an early sensitive marker for the diagnosis before the stage of microalbuminuria. and for predicting disease progression to start therapy very early.

Background: Breast cancer is the most common malignancy in Egyptian females. A wealth of literature is available on various diagnostic and prognostic parameters including clinical, pathological and biological factors. The present study focuses on markers of cell cycle proteins as cyclin B1 and parameters of cell cycle kinetics, related growth factors and tissue polyamines in relation to hormone receptor status.Patients and methods: A total of 102 females made the subject of the study, 72 females with breast cancer, 14 with benign lesions and 16 healthy subjects serving as controls for serum arameters. Normal breast tissue served as control for breast cancer tissues. Tissue polyamines were measured by HPLC. yclin B1 was measured by Flow cytometry (FCM) and Western Blotting (WB). Insulin-like Growth factor 1(IGF1) was easured by enzyme immunoassay (EIA). Tumor markers (CEA and CA 15.3), estradiol and progesterone levels were measured by microparticle enzyme immune assay (MEIA).Results: Cyclin B1 was detected in 17/34 (50%) of the cases tested by FCM and only in 10% of the cases evaluated by WB. Cyclin B1 positivity tended to be associated with high grade tumors. The mean percentage of cyclin B1 positivity was higher in aneuploid tumors than in diploid ones, as well as in tumors with ≥ 10% S-phase fraction (SPF) compared to those with < 10 % SPF. Cyclin B1 positivity was significantly associated with patients of high G2/M phase (p=0.05). Cell cycle analysis showed that ER-ve tumors were significantly associated with high SPF. Tissues polyamines showed significantly increased levels when compared to both benign and normal groups (p< 0.001). Polyamines showed a significant association with ER status (p= 0.04), tumor size (p=0.04) and histopathologic type (p= 0.05). Serum IGF 1 showed insignificant increased level in breast cancer patients less than 45 yrs when compared to the age matched control group. Serum estrogen level showed no change in the breast cancer patients ≥ 45yrs compared to age matched control group, while progesterone level was significantly lower (p< 0.02) in breast cancer patients less than 45yrs old compared to age matched benign and control groups. Serum CEA and CA 15.3 showed significant increase in the breast cancer patients < 45yrs old compared to age matched benign and control groups (p=0.03 and 0.06, respectively).Conclusion: Polyamines proved value as markers of proliferative activity of breast cancer but their testing needs further standardization. Cyclin B1 is associated with aggressive tumors. Its assessment by FCM proved superior to western blotting. IGF1 might be better used to assess tumor recurrence and monitor cancer patients, and its use needs further evaluation. This study may contribute to understanding breast cancer cell growth biology in Egyptian females in view of improving therapeutic and chemopreventive strategies.

Breast cancer (BC) is the most common neoplasm among women in most developed countries, including Egypt. Elevated levels of certain proteins in human BC are associated with unfavorable prognosis and progressive stages of the disease. The aim of our study was to evaluate the protein expression profile and prognostic significance of cyclooxygenase-2 (COX-2), matrix metalloproteinase-2 (MMP-2), MMP-9 and membrane type 1-MMP (MT1-MMP) and their interaction in operable BC patients. The protein expression of COX-2, MMP-2 and MT1-MMP were evaluated by western blot technique, whereas enzymatic activity of MMP-2 and MMP-9 was determined by zymography in 47 breast cancer patients as well as normal adjacent tissues. Also, the correlation between these proteins and age, tumor size, LN stage, TNM stage, estrogen receptor, progesterone receptor, disease-free survival, and overall survival (OS) has been investigated. As compared to adjacent normal tissues, COX-2, MMP-2 and MT1-MMP were over-expressed in 43, 64, and 60 % of tumor tissues, respectively. In the same pattern, the activity of MMP-2 (62 %) and MMP-9 (45 %) was elevated in BC tissues. Multivariate analysis showed a positive correlation between the protein expression of COX-2, MMP-2, and MT1-MMP and the activity of MMP-2 and MMP-9 in BC patients. However, the enzymatic activity showed no correlation with clinicopathological features. This study confirms the preclinical evidence that COX-2 increased the expression of MT1-MMP, which in turn activates MMP-2. The lack of correlation with clinicopathological features, OS or disease-free survival ascertains the complexity of tumor progression and metastasis with many pro- and counter regulatory factors.

Background and Aim: Response to interferon therapy and disease progression in hepatitis C virus (HCV) infected patients differs among individuals, suggesting a possibility of a contribution of host genetic factors. 2′‐5′‐oligoadenylate synthetase 1 (OAS1), an important component of the innate immune system with a proven antiviral function, may therefore have a relationship with the response to interferon therapy and clinical course of HCV disease. Our aim was to determine the frequency of single nucleotide polymorphism (SNP) at exon 7 splice acceptor site (SAS) of the OAS1 gene in relation to the interferon response and status of HCV infection. Methods: A 203 bp fragment containing exon 7 SAS was amplified in 70 HCV chronic patients and 50 healthy controls. SNP was examined using restriction fragment length polymorphism (RFLP) genotyping method. Correlations of SNP genotypes with response to interferon and clinical status of patients were statistically analyzed. Results: There was an increasing trend of response from AA to AG to GG genotypes (P = 0.007). Genotype AA was associated with non‐response to interferon and higher degree of liver fibrosis (P = 0.05). Multivariate analysis showed this SNP as independent and a significant determinant of the outcome of interferon therapy (odds ratio 4.913 [95% confidence interval 1.365–8.2], P = 0.006). Conclusions: This is the first study to show a significant association between the functional SNP at exon 7 SAS of OAS1 gene and the viral response to interferon in chronic HCV patients. Patients with AA genotype were associated with progressive HCV disease and viral resistance to interferon therapy. This OAS SNP is a potential bio‐marker to predict IFN response in chronic hepatitis C patients.

A post-prandial increase in saturated fatty acids (SFAs) and glucose (Glc) activates an inflammatory response, which may be prolonged following restoration of physiological SFAs and Glc levels — a finding referred to as ‘metabolic memory'. This study examined chronic and oscillating SFAs and Glc on the inflammatory signalling pathway in human adipose tissue (AT) and adipocytes (Ads) and determined whether Ads are subject to “metabolic memory.” Abdominal (Abd) subcutaneous (Sc) explants and Ads were treated with chronic low glucose (L-Glc): 5.6 mM and high glucose (H-Glc): 17.5 mM, with low (0.2 mM) and high (2 mM) SFA for 48 h. Abd Sc explants and Ads were also exposed to the aforementioned treatment regimen for 12-h periods, with alternating rest periods of 12 h in L-Glc. Chronic treatment with L-Glc and high SFAs, H-Glc and high SFAs up-regulated key factors of the nuclear factor-κB (NFκB) pathway in Abd Sc AT and Ads (TLR4, NFκB; P

Background Emerging data indicate that gut-derived endotoxin may contribute to low-grade systemic inflammation in insulin resistant states. This study aimed to examine the importance of serum endotoxin and inflammatory markers in non-alcoholic fatty liver disease (NAFLD) patients, with and without type 2 diabetes mellitus (T2DM), and to explore the effect of treatment with a lipase inhibitor, Orlistat, on their inflammatory status. Methods Fasted serum from 155 patients with biopsy proven NAFLD and 23 control subjects were analysed for endotoxin, soluble CD14 (sCD14), soluble tumour necrosis factor receptor II (sTNFRII) and various metabolic parameters. A subgroup of NAFLD patients were re-assessed 6 and 12 months after treatment with diet alone (n = 6) or diet plus Orlistat (n = 8). Results Endotoxin levels were significantly higher in patients with NAFLD compared with controls (NAFLD: 10.6(7.8, 14.8) EU/mL; controls: 3.9(3.2, 5.2) EU/mL, p < 0.001); NAFLD alone produced comparable endotoxin levels to T2DM (NAFLD: T2DM: 10.6(5.6, 14.2) EU/mL; non-diabetic: 10.6(8.5, 15.2) EU/mL), whilst a significant correlation between insulin resistance and serum endotoxin was observed (r = 0.27, p = 0.008). Both sCD14 (p < 0.01) and sTNFRII (p < 0.001) increased with severity of fibrosis. A positive correlation was also noted between sTNFRII and sCD14 in the NAFLD subjects (r = 0.29, p = 0.004). Sub-cohort treatment with Orlistat in patients with NAFLD showed significant decreases in ALT (p = 0.006), weight (p = 0.005) and endotoxin (p = 0.004) compared with the NAFLD, non-Orlistat treated control cohort at 6 and 12 months post therapy, respectively. Conclusions Endotoxin levels were considerably increased in NAFLD patients, with marked increases noted in early stage fibrosis compared with controls. These results suggest elevated endotoxin may serve as an early indicator of potential liver damage, perhaps negating the need for invasive liver biopsy. As endotoxin may promote insulin resistance and inflammation, interventions aimed at reducing endotoxin levels in NAFLD patients may prove beneficial in reducing inflammatory burden.

Background Matrix Metalloproteinases (MMPs) are key molecules for tumor growth, invasion and metastasis. Over-expression of different MMPs in tumor tissues can disturb the homeostasis and increase the level of various body fluids. Many MMPs including high molecular weights (HMWs) were detected in the urine of prostate and bladder cancer patients. Our aim here is to assess the usefulness of HMW MMPs as non invasive biomarkers in bilharzial bladder cancer in Egyptian patients. Methods The activity of different MMPs including HMW species was determined using zymographic analysis technique in the urine samples procured from sixty six bladder cancer patients (bilharzial and non-bilharzial) as well as hundred healthy control subjects. Also, the correlation between these HMW MMPs activities and different clinico-pathological parameters was investigated. Results High frequency of urine MMPs (uMMPs) activity was determined in 63.6% of examined tumor cases, however, none of the control cases showed any uMMPs activity. MMP-9 had the highest activity (62%) followed by MMP9/NGAL (60%), MMP-2 (54.5%), MMP-9 dimer (53%), ADAMTS (25.6%), and the lowest one was MMP-9/TIMP-1 (12%) only. There was no correlation between uMMPs and any of clinico-pathological parameters including age, gender, tumor size and type, bilharziasis, grade, lymph node involvement, and invasion to the prostate. A significant correlation was established only between MMP-9/TIMP-1 activities with the tumor size. Conclusions This study revealed that the detection of urinary MMPs including HMWs activity might be sensitive biomarkers for prediction of bladder cancer. It is also demonstrate that the detection of these urinary HMW gelatinases could not differentiate between bilharzial and non bilharzial bladder cancer subtypes.