Thymoquinone suppression of the human hepatocellular carcinoma cell growth involves inhibition of IL-8 expression,elevated levels of TRAIL receptors,oxidative stress and apoptosis

Sod2 is the major salt tolerance plasma membrane protein of Schizosaccharomyces pombe. It functions to remove excess intracellular sodium (or lithium) in exchange for protons. We investigated the role of cysteine residues and created a cysteine-free Sod2 protein. Each cysteine residue of the ten present was individually mutated to serine and the different proteins expressed and characterized in S. pombe. Western blotting revealed that all the individual mutant proteins were expressed. We examined the ability of the mutant proteins to confer salt tolerance to S. pombe with the endogenous Sod2 protein deleted. Only proteins with C26S and C374S mutations were partially reduced in their ability to confer salt tolerance. Additionally, they showed a change in conformation in comparison to the wild-type protein, indicated by differential sensitivity to trypsin. Deletion of all the cysteine residues of Sod2 resulted in a functional protein that was expressed in S. pombe at levels similar to the wild type and also conferred salt tolerance. The conformation of the cysteine-free Sod2 protein was not altered relative to the wild-type protein. We examined the accessibility of amino acids of the cysteineless protein present on putative extracellular loop 2. A cysteine placed at position Ala119 was accessible to externally applied [2-(trimethylammonium)ethyl] methane thiosulfonate bromide. The results demonstrate that cysteines in the Sod2 protein can be changed to serine residues resulting in an expressed, functional protein. The utility of the cysteine-free Sod2 protein for determination of topology and amino acid accessibility is demonstrated.     

Aneugenic Effects of Epirubicin in Somatic and Germinal Cells of Male Mice

The ability of the antineoplastic agent epirubicin to induce aneuploidy and meiotic delay in the somatic and germinal cells of male mice was investigated by fluorescence in situ hybridization assay using labeled DNA probes and BrdU-incorporation assay. Mitomycin C and colchicine were used as positive controls for clastogen and aneugen, respectively, and these compounds produced the expected responses. The fluorescence in situ hybridization assay with a centromeric DNA probe for erythrocyte micronuclei showed that epirubicin is not only clastogenic but also aneugenic in somatic cells in vivo. By using the BrdU-incorporation assay, it could be shown that the meiotic delay caused by epirubicin in germ cells was approximately 48 h. Disomic and diploid sperm were shown in epididymal sperm hybridized with DNA probes specific for chromosomes 8, X and Y after epirubicin treatment. The observation that XX- and YY-sperm significantly prevailed over XY-sperm indicates missegregation during the second meiotic division. The results also suggest that earlier prophase stages contribute less to epirubicin-induced aneuploidy. Both the clastogenic and aneugenic potential of epirubicin can give rise to the development of secondary tumors and abnormal reproductive outcomes in cured cancer patients and medical personnel exposed to epirubicin.     

Effect of Selenium and Vitamin C on Clinical Outcomes, Trace Element Status, and Antioxidant Enzyme Activity in Horses with Acute and Chronic Lower Airway Disease. A Randomized Clinical Trial

Excess production of reactive oxygen species is involved in the pathogenesis of airway disorders in horses. Trace element antioxidants have a beneficial role in oxidant/antioxidant balance. The aim of the present study was to evaluate the effect of a combination of sodium selenite and ascorbic acid on clinical outcome, antioxidant enzymes, and trace elements status in horses with lower airway disease. For this purpose, 40 draft horses with lower airway disease were randomly selected (acute, n?=?20; chronic, n?=?20). Both acute and chronic cases were randomly allocated into two subgroups (ten each). Groups 1 and 2 were the horses with acute disease, while groups 3 and 4 were chronically ill. For all groups, each horse was administered antibiotic, non-steroidal anti-inflammatory, and mucolytic drug. In addition, groups 2 and 4 were injected with 15 mg/kg sodium selenite and 30 mg/kg ascorbic acid every 24 h for successive 4 weeks. Venous blood samples were obtained from diseased horses on three occasions; at first examination, and at 2 and 4 weeks post-treatment. Clinically, antioxidant supplementation improved the clinical signs with significant decrease (p?<?0.05) of the clinical index score in both acute and chronic cases. In supplemented groups compared with non-supplemented, there was a significant increase (p?<?0.05) in the levels of copper, zinc, selenium, and iron as well as in the activity of glutathione-S-transferase and catalase. Meanwhile, there was a significant decrease (p?<?0.05) in the levels of manganese, malondialdehyde, hydrogen peroxide, and low-density lipoprotein and in the activity of glutathione reductase. The results of the present study indicate that administration of sodium selenite and ascorbic acid may have beneficial effect on clinical outcome and antioxidant balance in horses with acute and chronic lower airway disease.     

A possible new approach for controlling root knot nematode,Meloidogyne incognita,infecting eggplant by certain insect growth regulators

Under greenhouse conditions, three insect growth regulators, buprofezin, flufenoxuron and pyriproxyfen, at field concentrations and other two lower concentrations were evaluated for controlling of root knot nematode, Meloidogyne incognita, infecting eggplant (Solanum melongena L.) cv. Baladi. Generally, the tested pesticides reduced the number of galls, egg masses, females and developmental stages in roots and juveniles in soil compared to untreated check. The percentages of reduction were positively proportional with the respective concentrations. Flufenoxuron at its highest concentration was the most effective pesticide in reducing galls, egg masses, females and developmental stages compared with pyriproxyfen and buprofezin. The percentages of reduction caused by flufenoxuron were 67.80, 69.57, 70.75 and 95.23% for the, respective nematode criteria, while buprofezin at the highest concentration was the most effective in reducing number of hatched juveniles in soil followed by flufenoxuron and pyriproxyfen. Data showed that the length and fresh and dry weights of root and shoot of egg plant were increased with the lowest concentrations than the highest ones.     

Biodiversity of moderately halophilic bacteria in hypersaline habitats in Egypt

Screening bacteria from different saline environments in Alexandria. Egypt, lead to the isolation of 76 Gram-negative and 14 Gram-positive moderately halophilic bacteria. The isolates were characterized taxonomically for a total of 155 features. These results were analyzed by numerical techniques using simple matching coefficient (SSM) and the clustering was achieved by the unweighed pair-group method of association (UPGMA). At 75% similarity level the Gram-negative bacteria were clustered in 7 phena in addition to one single isolate, whereas 4 phena represented the Gram-positive. Based on phenotypic characteristics, it is suggested that the Gram-negative bacteria belong to the genera Pseudoalteromonas, Flavobacterium, Chromohalobacter, Halomonas and Salegentibacter, in addition to the non-identified single isolate. The Gram-positive bacteria are proposed to belong to the genera Halobacillus, Salinicoccus, Staphylococcus and Tetragenococcus. This study provides the first publication on the biodiversity of moderately halophilic bacteria in saline environments in Alexandria, Egypt.     

Enzymatic synthesis of γ-glutamylglutamine,a stable glutamine analogue,by γ-glutamyltranspeptidase from <Emphasis Type="Italic">Escherichia coli</Emphasis> K-12

The optimal reaction conditions for the synthesis of γ-glutamylglutamine using γ-glutamyltranspeptidase from Escherichia coli were determined. The maximum yield of γ-glutamylglutamine (110 mM) was obtained using 250 mM l-glutamine and 1.1 U γ-glutamyltranspeptidase/ml at pH 10.5 and at 37°C for 7 h; the conversion of glutamine to γ-glutamylglutamine was 88%.     

The Harderian gland of the Dhub lizard Uromastyx microlepis of the Kuwaiti desert: an ultrastructural approach

Harderian glands exist in the orbits of most terrestrial vertebrates. The basic function of the gland is the lubrication of the eye. The present study was carried out to shed some light on the ultrastructure of the still enigmatic Harderian gland of the lizard Uromastyx microlepis, a common species in Kuwait and other Gulf areas. The Harderian gland of Uromastyx microlepis is well developed, relatively large in size and lingual in shape. The epithelial cells of the anterior part of the gland are characterized by the presence of membrane bound granules of almost homogeneous consistency. These secretory granules are gathered in compartments and separated by membranes and stacks of granular endoplasmic reticulum (GER). Most of the lumina were empty. Moderate amounts of GER, free ribosomes and pleiomorphic mitochondria were observed in the perinuclear area of the epithelial cells. The medial and caudal parts of the gland were rich in special secretory granules, GER, free ribosomes and pleiomorphic mitochondria. The anterior part of the gland could represent the future lacrimal gland of mammals. A network of myoepithelial cells was recognized around the gland tubules. While no melanocytes or lymphocytes were observed in the scarce interstitial tissue, macrophages, that might have an immune function in the gland, were observed.     

Flavonoid glycosides of Artemisia monosperma and A. herba-alba

Ten flavonoid glycosides were isolated and identified from Artemisia monosperma: vicenin-2, lucenin-2, acacetin 7-glucoside, acacetin 7-rutinoside, the 3-glucosides and 3-rutinosides of quercetin and patuletin, and the 5-glucosides of quercetin and isorhamnetin. From Artemisia herba-alba eight flavonoid glycosides were isolated and identified: isovitexin, vicenin-2, schaftoside, isoschaftoside and the 3-glucosides and 3-rutinosides of quercetin and patuletin.     

Production of α-mannanase by B. subtilis from agro-industrial by-products: screening and optimization

We have demonstrated that a mixture of wheat bran (35 g l^-1), as a main substrate, and palm seed powder (10 g l^-1), as a co-substrate, is appropriate for -mannanase production by Bacillus subtilis. A 2ⁿ factorial experimental design was employed as a primary step for medium optimization. The enzyme activity titters obtained at the optimized growth condition were equivalent to about 319% of the -mannanse activity and 114% of the specific activity levels reached by a galactomannan-based culture.     

Alleviation of Aflatoxin B1‐Induced Genomic Damage by Proanthocyanidins via Modulation of DNA Repair

In order to study the mechanisms underlying the alleviation of aflatoxin B1‐induced genomic damage by proanthocyanidins (PAs), we examined the modulation of oxidative DNA damage induced by aflatoxin B1 in PAs‐pretreated animals. The effects of PAs on changes in the expression of DNA damage and repair genes induced by aflatoxin B1 were also evaluated in rat marrow cells. Administration of PAs before aflatoxin B1 significantly mitigated aflatoxin B1‐induced oxidative DNA damage in a dose‐dependent manner. Aflatoxin B1 treatment induced significant alterations in the expression of specific DNA repair genes, and the pre‐treatment of rats with PAs ameliorated the altered expression of these genes. Conclusively, PAs protect against aflatoxin B1‐induced oxidative DNA damage in rats. These protective effects are attributed to the antioxidant effects of PA and enhanced DNA repair through modulation of DNA repair gene expression. Therefore, PAs are a promising chemoprotective agent for averting genotoxic risks associated with aflatoxin B1 exposure.