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Direct acting antivirals against hepatitis C virus (HCV) have markedly improved cure rates in the past few years. However, they are expensive, with only few targeting host cell factors, and affecting virus assembly and release. Huh7.5 cells infected with a JFH-1 clone of HCV were treated with two different glycogen synthase kinase (GSK3)-β inhibitors; AR-A014418 and lithium chloride. Intra- and extracellular HCV virions and specific infectivity was determined using real-time RT-PCR and TCID50, and changes in lipid production were identified by enzyme-linked immunoassay and mass spectrometry analyses. Similarly, effect on two HCV replicon cells were identified by the luciferase activity. Although there was limited effect on virus replication in Huh7.5 cells and replicons, Huh7.5 cells treated with GSK3β inhibitors produced significantly less viral particles in comparison to untreated cells. In addition, the treated cells synthesized significantly lower amounts of ApoB and trapped the ApoE lipoproteins in the cells. In conclusion, our study suggests that GSK3β plays a pivotal role in HCV virion assembly and release mediated in part through inhibition of apolipoprotein synthesis.

Article History Tea is the second most popular drink in the world after water, and many studies have highlighted the effects of drinking tea. Therefore, the aim of this study was to explore the effects of white tea extract on the antioxidant enzymes activity including superoxide dismutase (SOD), glutathione peroxidase (GSH-px), and catalase (CAT). A significant decrease was observed in serum and liver SOD, GSH-px, and CAT activities in the diabetic control rats compared with the nondiabetic control ones. However, a significant increase in SOD, GSH-px, and CAT activities (serum and liver) was observed in the diabetic groups treated with white tea extract or Glibenclamide as compared to the diabetic control group. In conclusion, the present findings indicated that white tea extract has an enhancing effect on the antioxidant enzyme activities (SOD, CAT, and GSH-px) in STZ-induced diabetic rats.

This study aimed to investigate the biochemical influence of broccoli and beet extracts on selected individual additives NaNO2 or sunset yellow treated rats, in addition to the gene expression of some antioxidant enzymes. Forty-two male rats were assigned to seven groups of six rats in each group. The control group was fed a diet without an additive for four weeks. Group (2) received NaNO2, groups (3) received NaNO2 co-administered with broccoli extract (4) NaNO2 co-administered with beet extracts, Group (5) received sunset yellow, Group (6) received sunset yellow co-administered with broccoli extract, and Group (7) received sunset yellow co-administered with beet extract, for four weeks. At the end of the experiment, blood, liver, kidney, and brain samples were taken for biochemical and/or molecular analysis. The mRNA expression of antioxidant enzymes was determined by reversing transcriptase-polymerase chain reaction (RT-PCR). The obtained results revealed that rats co-administered with beet or broccoli extracts had a significant decrease in serum levels of AST, ALT, ALP, urea, total lipids, and triglycerides, as well as a significant increase in reduced glutathione (GSH), glutathione peroxidase (GSH-px), and superoxide dismutase (SOD) enzyme activities, compared to the normal control group. Oral administration of NaNO2 or sunset yellow caused a significant increase in serum levels of AST, ALT, ALP, urea, total lipids, and triglycerides, as well as a significant decrease in GSH, GSH-px, and SOD compared to the positive group. In conclusion, this study showed that broccoli and beet extracts have a protective effect against NaNO2 or sunset yellow in rat treated groups.

Article History Three new moderately thermophilic bacteria were isolated from three (Al-Khawbah, Bani Malik, and Al- Bozah) hot springs located in Jazan district, south-western of Saudi Arabia. The isolates were identified by the sequence analysis of the 16S rRNA gene. The 16S rRNA gene sequence alignment with GenBank resulted in a percent identity (% ID) with closest GenBank match of 98% suggesting that the isolates are new strains within the branching of well-defined genus Bacillus. The phylogenetic analysis of these strains using their 16S rDNA sequence data showed that strains SA and SS30 had highest homology (98%) with B. licheniformis, whereas strain SA09 showed 98% similarity with B. subtilis. The morphological, biochemical, and physiological characteristics of the isolates were also studied. They were aerobic, gram-positive, rod-shaped, and moderately thermophilic bacteria (with an optimum growth temperature of 45-50˚C, and pH of 6).

Staphylococcus hominis is a Gram-positive , spherical cells in clusters and coagulase -negative bacterial. It commonly occurs as a harmless commensal on human skin, occasionally causes nosocomial or community acquired infection specially in immuno-compromised patients. Nowadays almost all Staphylococcus species have multidrug resistance. In this study, a total of ten out of one hundred fifty nasal swabs at Assir Central Hospital General Lab during the period of April 2011- July 2011 proved to be Staphylococcus spp. and identified as Staphylococcus hominis patients of different sex and age groups with variable systemic infections ( e.g . RTI, UTI, CNS). The samples were tested by bactech, culture media, antibiotics sensitivity using diffusion disc test (MIC) and molecular polymerase chain reaction ( PCR) for confirmation of Staphylococcal species and detection of the Mec A gene. Clinical, demographic and laboratory data were collected and analyzed by SPSS. Drugs found to be resistant to all patients were penicillin, erythromycin, ampicillin, cifoxine and carbinicillin. Whereas cotrimexazole, amikacine and vancomycine were sensitive to all patients. Only 10% of patients were sensitive to methotrexate and cefaclor. Drugs that showed variable sensitivity pattern among patients were tetracyclin, fucidin, augmentin, gentamycin and ciprofloxacin. © 2013 JSR Publications. ISSN: 2070-0237 (Print); 2070-0245 (Online). All rights reserved. doi: http://dx.doi.org/10.3329/jsr.v5i1.11704 J. Sci. Res. 5 (1), 171-183 (2013)

Surface display technology can be defined as that phenotype (protein or peptide) which is linked to a genotype (DNA or RNA) through an appropriate anchoring motif. A bacterial surface display system is based on expressing recombinant proteins fused to sorting signals (anchoring motifs) that direct their incorporation on the cell surface.

Background: The oral delivery of proteins and peptide drugs is considered a major challenge. These types of therapeutics are readily degraded, if taken orally, due to the harsh high acidity of stomach and enzymatic attack in the upper small intestinal tract. Methods: Water-soluble copolymers of sodium acrylate (AAs) grafted onto carboxymethyl cellulose (CMC) were prepared and characterized using Fourier transform spectroscopy, differential scanning calorimetry, and X-ray diffraction. The obtained graft copolymers were then used in a combination with sodium alginate to develop a new series of pH-sensitive interpenetrating polymeric network (IPN) hydrogels through ionotropic gelation with divalent ions (Ca 2+ ). Morphology of the developed hydrogels was investigated using SEM. Swelling characteristics, at distinct compositions, were also studied at 37°C in two consecutive buffer solutions of pH 2.1 and 7.4 (similar to that of gastric and intestinal fluids, respectively). The release profiles of bovine serum albumin, as a model protein, from test IPN hydrogel films were studied in simulated gastric and intestinal fluids. In addition, the drug release process was confirmed by means of SEM. Results: Swelling studies of the developed IPN hydrogels at different pH values confirmed their pH-sensitive nature. The equilibrium swelling extents of the hydrogels were found to be dependent on the grafting yield of CMC/AAs graft copolymer. The IPN hydrogels attained equilibrium swelling percentages in the range 445–740%. In addition, the amount of bovine serum albumin released within 2 hours in pH 2.1 was relatively low (less than 18.1%). This amount increased up to 68% after 8 hours in pH 7.4. Conclusions: From the obtained preliminary data, it seems that the IPN hydrogels developed in this contribution can be tailored to act as good potential carriers for oral delivery of protein drugs. These hydrogels showed a promising protection of protein drugs from the harsh acidity of stomach and, at the same time, they conferred sustained drug release in the intestinal fluid.

Using an EDC/NHS conjugation method, chitosan was chemically modified to incorporate a photosensitive α-cyano-4-hydroxycinnamic acid moiety with various degrees of substitutions. The chitosan α-cyano-4-hydroxycinnamate was fully characterized by FTIR, 1 H NMR and UV–vis spectra. Lipase from Candida rugosa was entrapped in the modified photo-crosslinkable chitosan membranes. Crosslinking was carried out by irradiation in the ultraviolet region. The activities of free and immobilized lipase have been studied. The efficiency of the immobilization was evaluated by examining the relative enzymatic activity of free enzyme before and after the immobilization of lipase. The obtained values were found to reach 98.6%. The results showed that the optimum temperature of immobilized lipase was 40 °C, which was identical to that of the free enzyme, and the immobilized lipase exhibited a higher relative activity than that of free lipase over 40 °C. The optimal pH for immobilized lipase was 8.0, which was higher than that of the free lipase (pH 7.5), and the immobilization resulted in stabilization of enzyme over a broader pH range. The kinetic constant value ( K m ) of immobilized lipase was 1.332 mg ml −1 (1.50 × 10 −3 M), which was higher than that of free lipase. On the other hand, the activity of immobilized lipase decreased slowly against time when compared to that of the free lipase, and could retain 75.5% residual activity after 6 consecutive cycles.

A method has been developed to immobilize horseradish peroxidase (HRP) on modified chitosan beads by means of graft copolymerization of polyethylacrylate in presence of potassium persulphate and Mohr's salt redox initiator. The activity of free and immobilized HRP was studied. FTIR spectroscopy and scanning electron microscopy were used to characterize HRP immobilization. The efficiency of the immobilization was investigated by examining the relative enzymatic activity of free enzyme before and after the HRP immobilization. The obtained values were found to reach 98.4%. The results show that the optimum temperature of immobilized HRP was 45 degrees C, which was identical to that of free enzyme, and the immobilized HRP exhibited a higher relative activity than that of free HRP over 45 degrees C. The optimal pH for immobilized HRP was 10, which was higher than that of the free HRP (pH 9.0), and the immobilization resulted in stabilization of enzyme over a broader pH range. The apparent kinetic constant value (K(m)) of immobilized HRP was 3.784 mmol ml(-1), which was higher than that of free HRP. On the other hand, the activity of immobilized HRP decreased slowly against time when compared to that of the free HRP, and could retain 65.8% residual activity after 6 consecutive cycles.

A method has been developed to immobilize lipase from Candida rugosa on modified natural wool fibers by means of graft copolymerization of poly ethylacrylate in presence of potassium persulphate and Mohr’s salt redox initiator. The activities of free and immobilized lipase have been studied. FTIR spectroscopy, scanning electron microscopy, and the Bradford method were used to characterize lipase immobilization. The efficiency of the immobilization was evaluated by examining the relative enzymatic activity of free enzyme before and after the immobilization of lipase. The results showed that the optimum temperature of immobilized lipase was 40 °C, which was identical to that of the free enzyme, and the immobilized lipase exhibited a higher relative activity than that of free lipase over 40 °C. The optimal pH for immobilized lipase was 8.0, which was higher than that of the free lipase (pH 7.5), and the immobilization resulted in stabilization of enzyme over a broader pH range. The kinetic constant value (km) of immobilized lipase was higher than that of the free lipase. However, the thermal and operational stabilities of immobilized lipase have been improved greatly.

Background: The genus Acinetobacter is a diverse group of Gram-negative bacteria involve at least 33 species using the molecular methods. Although the genus Acinetobacter comprises a number of definite bacterial species, some of these species are of clinical importance. Therefore, it is of vital importance to use a method which is able to reliably and efficiently differentiate the numerous Acinetobacter species. Objectives: This study aims to identify Acinetobacter of clinical isolates from Assir region to the species level by 16S-23S intergenic spacers internal transcribed spacer (ITS) of ribosomal ribonucleic acid (rRNA). Materials and Methods: Deoxyribonucleic acid extraction, polymerase chain reaction amplification of 16S-23S intergenic spacer sequences (ITS) was performed using the bacterium-specific universal primers. Results: Based on the 16S-23S intergenic spacers (ITS) of rRNA sequences, all isolates tested were identified as Acinetobacter baumannii. The isolates shared a common ancestral lineage with the prototypes A. baumannii U60279 and U60280 with 99% sequence similarities. Conclusion: These findings confirmed 16S-23S rRNA ITS for the identification of A. baumannii of different genotypes among patients.

Hepatitis C virus (HCV) is a hepatotropic virus that also infects cells of the immune system. HCV clones cultivated in human hepatoma Huh-7.5 cells have significantly advanced our understanding of HCV replication and candidate hepatocyte receptors. However, naturally occurring patient-derived HCV, in contrast to the HCV JFH-1 clone, is unable to infect Huh-7.5 cells, while it can replicate in human primary T-cells and selected T-cell lines. To better understand this incongruity, we examined the susceptibility of primary T-cells, PBMCs and T-cell lines to infection with patient-derived HCV, the classical HCV JFH-1 and a cell culture-adapted JFH1T known to be highly infectious to Huh-7.5 cells. We also tested whether Huh-7.5 cells are prone to virus readily infecting T-lymphocytes. The results revealed that while primary T-cells and Molt4 and Jurkat T-cell lines were susceptible to patient-derived HCV, they were resistant to infection with either JFH1T or JFH-1. However, the JFH1T clone interacted more firmly, although non-productively, with the cells than JFH-1. Further, Huh-7.5 cells robustly supported replication of JFH1T but not patient-derived, wild-type virus, despite using highly sensitive detection assays. In conclusion, JFH-1 and JFH1T clones were unable to establish productive infection in human primary T-cells, PBMCs and T-cell lines known to be prone to infection by patient-derived HCV, while Huh-7.5 cells were resistant to infection with naturally occurring virus infecting immune cells. The data showed that the ability to infect lymphocytes is a characteristic of native virus but not laboratory HCV clones.

Hepatitis C virus (HCV) is one of the main causes of chronic liver disease. Although infection of hepatocytes is mainly responsible for manifestations of hepatitis C, the virus also invades the immune system by a yet-to-be-identified mechanism. Using human T cell lines and primary T lymphocytes as targets and patient-derived HCV as inocula, we aimed to identify how HCV gains entry into these cells. HCV replication was determined by detection of the HCV RNA replicative (negative) strand and viral proteins, while specific antibodies, knocking down gene expression and making otherwise-resistant cells prone to HCV, were employed to identify a receptor molecule determining T lymphocyte permissiveness to HCV infection. The results revealed that T cell susceptibility to HCV requires CD5, a lymphocyte-specific glycoprotein belonging to the scavenger receptor cysteine-rich family. Blocking of T cell CD5 with antibody or silencing with specific short hairpin RNA (shRNA) decreased cell susceptibility to HCV, while increasing CD5 expression by mitogen stimulation had the opposite effect. Moreover, transfection of naturally CD5-deficient HEK-293 fibroblasts with CD5 facilitated infection of these otherwise HCV-resistant cells. In contrast to T cells, hepatocytes do not express CD5. The data revealed that CD5 is a molecule important for HCV entry into human T lymphocytes. This finding provides direct insight into the mechanism of HCV lymphotropism and defines a target for potential interventions against HCV propagating in this extrahepatic compartment.

Accumulated evidence implies that hepatitis C virus (HCV) infects not only the liver but also the immune system. A lymphocyte-specific CD5 molecule was recently identified as essential for infection of T cells with native, patient-derived HCV. To assess whether the proposed hepatocyte receptors may also contribute to HCV lymphotropism, expression of scavenger receptor-class B type 1 (SR-B1), claudin-1 (CLDN-1), claudin-6 (CLDN-6), occludin (OCLN), CD5 and CD81 was examined by real-time RT-PCR and the respective proteins quantified by immunoblotting in HCV-prone and resistant T cell lines, peripheral blood mononuclear cells (PBMC), primary T cells and their subsets, and compared to hepatoma Huh7.5 and HepG2 cells. SR-B1 protein was found in T and hepatoma cell lines but not in PBMC or primary T lymphocytes, CLDN-1 in HCV-resistant PM1 T cell line and hepatoma cells only, while CLDN-6 equally in the cells investigated. OCLN protein occurred in HCV-susceptible Molt4 and Jurkat T cells and its traces in primary T cells, but not in PBMC. CD5 was displayed by HCV-prone T cell lines, primary T cells and PBMC, but not by non-susceptible T and hepatoma cell lines, while CD81 in all cell types except HepG2. Knocking-down OCLN in virus-prone T cell line inhibited HCV infection, while de novo infection downregulated OCLN and CD81, and upregulated CD5 without modifying SR-B1 expression. Overall, while no association between SR-B1, CLDN-1 or CLDN-6 and the susceptibility to HCV was found, CD5 and CD81 expression coincided with virus lymphotropism and that of OCLN with permissiveness of T cell lines but unlikely primary T cells. This study narrowed the range of factors potentially utilized by HCV to infect T lymphocytes amongst those uncovered using laboratory HCV and Huh7.5 cells. Together with the demonstrated role for CD5 in HCV lymphotropism, the findings indicate that virus utilizes different molecules to enter hepatocytes and lymphocytes.

Thiocyclam (trade name Evisect) is a broad-spectrum nereistoxin analogue insecticide used widely for agricultural applications. The aim of this investigation was to determine its genotoxic effects in the chromosome aberration (CA) test and determining of mitotic index (MI), using lymphocytes from peripheral blood samples of healthy human donors. A negative and a positive control (MMC) were also included. Chromosomal analyses of the metaphase plates of the samples treated with 14 different concentrations (from 0.1 to 120μg/ml) of thiocyclam, indicating the lack effect on chromosomes. Thus thiocyclam is not genotoxic but highly toxic on cell proliferation in human lymphocytes.

Glutathione S-transferases have been purified (18-fold) in 65-70% yield from the liver of one humped camel using affinity chromatography on glutathione-linked agarose. Chromatofocusing technique resolves the glutathione S-transferases into seven distinct isoenzymes with apparent pI of 8.7, 8.4, 8.0, 7.8, 7.3 and 6.5. The major isoenzyme (pI 8.7) which accounted for over 95% of the total activity was composed of two identical subunits of molecular mass 24,000 and was immunologically similar to the other six isoenzymes. The substrate specificities and the effect of various inhibitors on the activity of the abundant camel liver isoenzyme were also examined.