THE PROTECTIVE EFFECT OF THYMOQUINONE AGAINST LEAD ACETATE INDUCED DNA DAMAGE AND ALTERATIONS IN TUMOR INITIATION GENES

objective: Several pollutants represent a significant ecological and public health concern due to their toxicity and their ability to accumulate in livingorganisms in which lead is one of them. The present investigation was designated to assess the modulating effect of thymoquinone (TQ) against leadacetate (LA) toxicity.Methods: Several endpoints were considered to design this study such as: The gene expression of tumor initiation genes (cytochrome P450 3A[CYP3A], cyclooxygenase 2 [COX2], BAX and Bcl), DNA damage and alterations in the levels of glutathione (GSH), lipid oxidation (malondialdehyde[MDA]), and protein oxidation (protein carbonyl [PC]) in male rats. About 60 male rats were used in this study which allocated in six groups (10 animaleach) and treated with LA (200 mg/kg diet), TQ (5 and 10 mg/kg b.wt.), and LA + TQ.2Results: The results revealed that LA induced significant DNA damage and alteration in the expression of CYP3A, COX2, BAX, and Bcl as well asinduced changes in GSH content and MDA and PC levels in male rats. Meanwhile, TQ was decreased significantly the toxic effect of LA in male ratswhich decreased the alterations in the gene expression and DNA damage as well as GSH content and MDA and PC levels.Conclusion: The results suggested that TQ treatment confers protection against toxicity inflicted by LA and support the contention that TQ protectionis achieved by its ability as a scavenger for free radicals generated by LA.Keywords: Thymoquinone, Lead acetate, Gene expression, DNA damage, Rats

Genetic polymorphism of five genes associated with growth traits in goat

Genetic polymorphism studies in domestic animals aim at evaluating genetic variations within and across breeds mainly for conservation purposes. In this study, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect polymorphisms of five candidate genes in four Egyptian and Saudi goat breeds (Barki, Zaribi, Ardi and Masri), to detect the genotype of GH, IGF1, POUIF1, MSTN and BMP15 genes in the goat breeds and their allele frequencies. Results of GH gene which encloses a Haelll endonuclease restriction site show four unique PCR-RFLP banding patterns (genotypes AA, AB, CC and CD). The frequencies of the A allele in the samples from the goat breeds varied from 0.410 to 0.620. While  IGF-1gene revealed three fragments after digestion with Haelll with genotype AA, AB and BB and the  frequencies of allele A varied from 0.432 to 0.731. Furthermore, PCR-RFLP of POUIF1 gene showed two  fragments after digestion by Pst1 endonuclease with genotype TT and CC and the frequencies of allele T varied from 0.250 to 0.840. The MSTN gene revealed three fragments after digestion with DraI with genotype AA, BB and AB and the frequencies of allele A varied from 0.240 to 0.630. Meanwhile, the BMP15 gene revealed one fragments of 112 bp for AA after digestion with Hinf1 enzyme.Key words: Goats, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), GH, IGF-1, POUIF1, MSTN, BMP-15

Metabolomic Response of Calotropis procera Growing in the Desert to Changes in Water Availability

Water availability is a major limitation for agricultural productivity. Plants growing in severe arid climates such as deserts provide tools for studying plant growth and performance under extreme drought conditions. The perennial species Calotropis procera used in this study is a shrub growing in many arid areas which has an exceptional ability to adapt and be productive in severe arid conditions. We describe the results of studying the metabolomic response of wild C procera plants growing in the desert to a one time water supply. Leaves of C. procera plants were taken at three time points before and 1 hour, 6 hours and 12 hours after watering and subjected to a metabolomics and lipidomics analysis. Analysis of the data reveals that within one hour after watering C. procera has already responded on the metabolic level to the sudden water availability as evidenced by major changes such as increased levels of most amino acids, a decrease in sucrose, raffinose and maltitol, a decrease in storage lipids (triacylglycerols) and an increase in membrane lipids including photosynthetic membranes. These changes still prevail at the 6 hour time point after watering however 12 hours after watering the metabolomics data are essentially indistinguishable from the prewatering state thus demonstrating not only a rapid response to water availability but also a rapid response to loss of water. Taken together these data suggest that the ability of C. procera to survive under the very harsh drought conditions prevailing in the desert might be associated with its rapid adjustments to water availability and losses

Gene expression and histopathology alterations during rat mammary carcinogenesis induced by 7,12- dimethylbenz[a]anthracene and the protective role of Neem (Azadirachta indica) leaf extract

Abstract: The present study was investigated to evaluate the protective role of ethanolic neem leaf extract (ENLE) against 7,12-dimethylbenz[a]anthracene (DMBA)-induced expression alterations of the Bcl-2, CK8, CK19, p53, p21, p27 and PCNA genes and histopathological lesions in the mammary tissues of female rats. Eighty Swiss albino female rats were divided into eight groups. Group 1 supplemented with corn oil as control. Group 2 females received DMBA. Groups 3, 4 and 5 females received 100, 200 and 400 mg/kg of ENLE alternate to the DMBA application from the beginning of the experimental period, respectively. Groups 6, 7, and 8 females treated similar to groups 3, 4 and 5 plus DMBA. All the animals were sacrificed after an experimental period of 12 weeks. The expression of Bcl-2, CK8, CK19, p53, p21, p27 and PCNA genes was investigated using reverse transcription polymerase chain reaction (RT-PCR). In addition, histopathology analysis of the mammary tissues was confirmed. The results revealed that DMBA treatment induced expression alterations of genes related to cancer. Also histopathological lesions were found in mammary tissues of female rats. These alterations of the gene expression as well as the histopathological lesions were markedly suppressed when female rats were treated with ENLE combined with DMBA. Conclusion: These findings suggest that ENLE exerts its anticancer properties by inhibiting alterations in the gene expression and the histopathological lesions in the mammary tissues of female rats exposed to DMBA

Functional differences between estrogen receptors alpha and beta in human breast cancer cells in culture

EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Effect of the purification of antidermatophytic proteins from Nigella sativa on four zoophilic species

The antidermatophytic activities of proteins which are extracted from four plant species (Carum carvi, Cymbopogon citratus, Moringa oleifera, and Nigella sativa) on four zoophilic dermatophytes (Microsporum canis, Microsporum equinum, Trichophyton mentagrophytes, and Trichophyton verrucosum) were evaluated in this study. The crude proteins of N. sativa had the broadest significant spectrum of antidermatophytic activity on the tested dermatophytes as well as the greatest antioxidant activity (95.11%) and the highest protein content (82 mg/ml). 17 amino acids were found in the four tested plant proteins with N. sativa protein having the highest content of amino acid (347.21 μg/ml). N. sativa protein had the greatest effect on the fungal cell permeability of all the tested zoophilic dermatophytes. Purification of N. sativa protein on Sephadex G-100 column showed two peaks of protein (Pr1 and Pr2) as well as increasing antidermatophytic activities. Complete purification of the most active fraction (Pr2) on diethylaminoethyl (DEAE)-Sephadex eluted one single peak with increasing antidermatophytic activity (7.6 cm) against the most sensitive pathogen (M. canis), representing 1.43 fold purification of the crude protein. The molecular weights of the purified N. sativa proteins (Pr1 and Pr2) were 42.7 and 31 kDa, respectively. The highest antidermatophytic activity of Pr2 was observed at a pH of 7 and temperature of 20°C. Na+, K+, Ca2+ and Mg2+ decreased the antidermatophytic activity of the pure protein of N. sativa.Key words: Dermatophytes, plant, protein, purification

Sequence analysis of sub-genotype D hepatitis B surface antigens isolated from Jeddah, Saudi Arabia

Little is known about the prevalence of HBV genotypes/sub-genotypes in Jeddah province, although the hepatitis B virus (HBV) was identified as the most predominant type of hepatitis in Saudi Arabia. To characterize HBV genotypes/sub-genotypes, serum samples from 15 patients with chronic HBV were collected and subjected to HBsAg gene amplification and sequence analysis. Phylogenetic analysis of the HBsAg gene sequences revealed that 11 (48%) isolates belonged to HBV/D while 4 (18%) were associated with HBV/C. Notably, a HBV/D sub-genotype phylogenetic tree identified that eight current isolates (72%) belonged to HBV/D1, whereas three isolates (28%) appeared to be more closely related to HBV/D5, although they formed a novel cluster supported by a branch with 99% bootstrap value. Isolates belonging to D1 were grouped in one branch and seemed to be more closely related to various strains isolated from different countries. For further determination of whether the three current isolates belonged to HBV/D5 or represented a novel sub-genotype, HBV/DA, whole HBV genome sequences would be required. In the present study, we verified that HBV/D1 is the most prevalent HBV sub-genotype in Jeddah, and identified novel variant mutations suggesting that an additional sub-genotype designated HBV/DA should be proposed. Overall, the results of the present HBsAg sequence analyses provide us with insights regarding the nucleotide differences between the present HBsAg/D isolates identified in the populace of Jeddah, Saudi Arabia and those previously isolated worldwide. Additional studies with large numbers of subjects in other areas might lead to the discovery of the specific HBV strain genotypes or even additional new sub-genotypes that are circulating in Saudi Arabia. Keywords: Hepatitis B virus, HBV sub-genotypes, HBV/D, HBsAg, Viral isolates, Population studie

Production and characterization of thermostable xylanase from Bacillus subtilis XP10 isolated from marine water

This study aims is to characterize an efficient bacterium, capable of producing thermostable xylanases. Different bacterial isolates were obtained on medium containing xylan as the sole carbon source from samples collected from Jeddah, Saudi Arabia. Out of 15 xylanase producing isolates, the best xylanase producer was XP10 which was selected for the enzyme production. It was identified based on morphological and some biochemical characters as a species belonging to the genus Bacillus and identified as Bacillus subtilis XP10. Identification was confirmed by 16S rDNA studies and restriction fragment length polymorphism. Factors affecting enzymes production were studied. Maximum xylanase production was 2.82 U/ml obtained at pH 8 for 4 days at 40°C and 120 rpm. The molecular weight of the purified enzyme was 23 kDa. Fivefold increasing in xylanase production was obtained by construction of recombinant Escherichia coli 107 harboring recombinant vector pJET 1.2/blunt/xynA. The purified thermostable alkalotolerant xylanase can be used in the treatment of agricultural wastes as well as in the bioremediation of xylan-containing materials.Keywords: Xylanase, polymerase chain reaction based restriction fragment length polymorphism (PCR-based RFLP), 16S rDNA, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE), xynA geneAfrican Journal of Biotechnology Vol. 12(8), pp. 780-79

<i>Actinidia deliciosa</i> Extract as a Promising Supplemental Agent for Hepatic and Renal Complication-Associated Type 2 Diabetes (In Vivo and In Silico-Based Studies)

Type 2 diabetes (T2D) is a chronic metabolic condition associated with obesity, oxidative stress-mediated inflammation, apoptosis, and impaired insulin signaling. The utilization of phytochemical therapy generated from plants has emerged as a promising approach for the treatment of diabetes and its complications. Kiwifruit is recognized for its substantial content of antioxidative phenolics. Therefore, this work aimed to examine the effect of Actinidia deliciosa (kiwi fruit) on hepatorenal damage in a high-fat diet (HFD) and streptozotocin (STZ)-induced T2D in rats using in vivo and in silico analyses. An increase in hepatic and renal lipid peroxidation was observed in diabetic rats accompanied by a decrease in antioxidant status. Furthermore, it is important to highlight that there were observable inflammatory and apoptotic responses in the hepatic and renal organs of rats with diabetes, along with a dysregulation of the phosphorylation levels of mammalian target of rapamycin (mTOR), protein kinase B (Akt), and phosphoinositide 3-kinase (PI3K) signaling proteins. However, the administration of kiwi extract to diabetic rats alleviated hepatorenal dysfunction, inflammatory processes, oxidative injury, and apoptotic events with activation of the insulin signaling pathway. Furthermore, molecular docking and dynamic simulation studies revealed quercetin, chlorogenic acid, and melezitose as components of kiwi extract that docked well with potential as effective natural products for activating the silent information regulator 1(SIRT-1) pathway. Furthermore, phenolic acids in kiwi extract, especially syringic acid, P-coumaric acid, caffeic acid, and ferulic acid, have the ability to inhibit the phosphatase and tensin homolog (PTEN) active site. In conclusion, it can be argued that kiwi extract may present a potentially beneficial adjunctive therapy approach for the treatment of diabetic hepatorenal complications