Walaa Awadin

Anticancer activity of Aloe vera and Calligonum comosum extracts separetely on hepatocellular carcinoma cells

ABSTRACT Objective To investigate the in vitro anticancer effect of Aloe vera ( A. vera ) and Calligonum comosum ( C. comosum ) extracts against hepatocellular carcinoma (HepG2) cells. Methods HepG2 cells were tested against different doses of A. vera and C. comosum . Viability of the cells was assessed by MTT assay. Evaluation of apoptosis and DNA damage in HepG2 cells were performed using annexin V apoptosis detection kit. The expression of p53 and anti-apoptotic (Bcl-2) were tested by real time-PCR and flow cytometer analyser. Hematoxylin and eosin stained sections from untreated and treated HepG2 cells were observed using light microscopy. Results The IC 50 values of A. vera and C. comosum extracts were (10.45 ± 0.31) and (9.60 ± 0.01) µg/mL respectively. The extracts separately increased cytotoxicity against HepG2 cells in a time and dose dependent manners. Also, it apparently induced apoptosis through increase P53 and decrease Bcl-2 genes expressions. Conclusions The results indicated that the extracts could have anti-hepatocarcinogenic effect, at least in part, through modulation of apoptosis.

Overproduction of Toxoplasma gondii cyclophilin-18 regulates host cell migration and enhances parasite dissemination in a CCR5-independent manner

BackgroundToxoplasma gondii hijacks host cells to allow it to disseminate throughout a host animal; however, the migratory machinery involved in this process has not been well characterized. We examined the functional role of T. gondii cyclophilin 18 (TgCyp18) in host cell recruitment using recombinant parasites transfected with TgCyp18.ResultsHigh levels of TgCyp18 enhanced IL-12 production in cysteine-cysteine chemokine receptor 5 (CCR5) knockout mice (CCR5−/−) that had been infected peritoneally with T. gondii. Recruitment of CD11b+ cells to the infection site was enhanced in a CCR5-independent manner. T. gondii spread to several organs, particularly the liver, in a TgCyp18-dependent and CCR5-independent manner. Additionally, CCL5 levels were upregulated in macrophages treated with recombinant protein TgCyp18 and in the peritoneal fluids of the infected CCR5−/− mice. Furthermore, the chemokines involved in macrophage migration, CCL2 and CXCL10, were upregulated in the livers of CCR5−/− mice infected with recombinant parasites that had been transfected with TgCyp18.ConclusionTgCyp18 may play a crucial role in macrophage migration, and in assisting with transport of T. gondii via CCR5-independent mechanisms. TgCyp18 may also play a role with CCL5 in the migration of macrophages to the site of infection, and with CCL2 and CXCL10 in the transport of T. gondii-infected cells to the liver.

Detection of Lumpy Skin Disease Virus Antigen and Genomic DNA in Formalin-Fixed Paraffin-Embedded Tissues from an Egyptian Outbreak in 2006

An outbreak of lumpy skin disease (LSD) was reported in 2006 in Egypt affecting 16 provinces. Biopsies and post-mortem tissue samples were collected from calves that showed typical clinical signs of LSD and fixed in formalin. These samples were collected from a private dairy farm in the Damietta province of Egypt. Formalin-fixed paraffin-embedded tissue samples were assessed using histology, and skin lesions were classified as either acute or subacute/chronic. Both lumpy skin disease virus (LSDV) DNA detected by polymerase chain reaction and LSDV antigen detected by immunohistochemistry using a capripoxvirus-specific monoclonal antibody were observed in the acute skin lesions and in some subacute/chronic skin lesions.

Cardioprotective role of tadalafil against cisplatin-induced cardiovascular damage in rats

The present study investigated the possible cardioprotective effect of tadalafil (Tad) on cisplatin (CDDP)-induced cardiac and vascular damages in rats. A total number of seventy two healthy male albino rats initially weighting between 200 and 220 g were used and randomly divided into four groups,18 rats in each. The control group received no treatment; CDDP group received a single dose of CDDP (4 mg/kg) intraperitoneal (i.p.) per week for 4 weeks the duration of the experiment; Tad group received 0.4 mg/kg BW Tad i.p. daily and Tad +CDDP group received 0.4 mg/kg BW Tad i.p. +4 mg/kg BW CDDP i.p. The results showed that Tad was able to decrease blood pressure, heart rate, levels of serum cardiac troponin (cTn-I), malondialdehyde (MDA) and increased levels of reduced glutathione (GSH) and nitric oxide (NO) in the heart homogenate sample from CDDP treated rats. Semi-quantitative analysis showed that Tad was able to decrease the histopathological scores of cardiac muscular hyalinzation and fibrosis in three sacrifices in CDDP treated rats. CDDP treated rats showed significantly increased thickening in wall of aorta with an irregular luminal layer of endothelial cell linings in three sacrifices when it was compared to other groups. Moreover, immunohistochemical labeling of α- smooth muscle actin (α-SMA) in aorta revealed significant lower scores in Tad +CDDP group when they were compared to CDDP group. In conclusion, Tad alone did not induce any harmful effects on blood pressure, selective antioxidant, peroxidation markers or cardiac histology, in addition, Tad has a cardio-protective role against CDDP.

Gentamicin-induced Acute KidneyInjury in Equines is associated withMarked Acute Phase Response: AnExperimental Study on Donkey(Equus asinus)

Gentamicin-induced Acute Kidney Injury in Equines is associated with Marked Acute Phase Response: An Experimental Study on Donkey (Equus asinus) Recently, there is growing evidence suggesting that acute kidney injury (AKI) in human and laboratory animals is associated with an inflammatory response that could play a role in tissue damage. However, such link has not previously been addressed in equines. The present study was designed to evaluate the effect of gentamicin (GEN) administration on the development of AKI and systemic inflammatory response in equines using donkey as a model. GEN (10%) was administered intravenously in six donkeys at a dose of 20 mg kg-1 BW thrice daily for 14 consecutive days. Three other donkeys were randomly assigned to receive saline solution and served as controls. The donkeys were clinically and sonographically examined throughout the experimental period. Blood and urine (U) samples were simultaneously collected at day (D) 7, and D 14 of GEN administration. Renal specimens from all donkeys were collected at D 14 and processed for routine histopathological examination. AKI was confirmed by sonography, laboratory measurements, histopathology and immunohistochemistry.

Acute exposure to chlorpyrifos induces reversible changes in health parameters of Nile tilapia (Oreochromis niloticus)

Chlorpyrifos (CPF) is one of the most common insecticides found in freshwater ecosystems, and has been detected in agricultural and fishery products worldwide. This study focused on comprehensive panel of hematological, immunotoxic and pathology changes in Nile tilapia (Oreochromis niloticus) during and after exposure to CPF at 15 μg/L (0.043 μM) (1/10 LC50, group CPF1), or 75 μg/L (0.21 μM) (1/2 LC50, group CPF2) for 14 days, followed by 2 weeks recovery. Different endpoints were used to determine effects of CPF on fish health: hematological parameters; antioxidant levels in liver and gills; innate immune parameters; expression levels of pro-inflammatory cytokine genes at mRNA level in anterior kidney and spleen; and histopathological assessment of gills, liver, and kidney tissues. RBCs were significantly decreased in CPF1 group compared to other groups only at day 3. Blood packed cell volume (PCV) and mean corpuscular volume (MCV) showed significant increase at day 3 and 14 of CPF exposure. TLC (Total Leukocytic Counts), neutrophil counts were significantly increased in CPF exposed groups at days 3, 7, 14 compared to the control. While, lymphocytes counts were significantly increased at CPF1 group compared to other groups at day 14. Antioxidant enzyme activity in liver and gills showed significant increase of Malondialdehyde (MDA) and glutathione (GSH), and significant decrease in (catalase/CAT/, glutathione S-transferase/GST/, and superoxide dismutase/SOD/); in CPF exposed groups. Serum bactericidal and lysozyme activity was nominally and significantly decreased, respectively, and whole blood respiratory burst was significantly increased in CPF2 group. The cytokine expression levels showed complex changes in expression patterns. In kidney, cytokine interleukin-8 (IL-8) was significantly upregulated at day 1 in both exposed group. Interleukin-1β (IL-1β) and Tumor necrosis factor-α (TNFα) were significantly upregulated at day 1 in CPF1 group, and then IL-8 and TNFα downregulated at day 3 in same group. At day 7, only TNFα was up and downregulated in CPF1 and CPF2, respectively compared to control. All gene expression levels in spleen were upregulated on day 7 of exposure in the high exposed group. Histopathology showed dose-dependent changes in CPF treated groups, indicating gill, liver, and posterior kidney changes associated with oxidative stress damages. Following recovery period, all measured parameters showed varying degrees in their reversibility to the control level. These findings provide important insights about the acute toxic effects of CPF on fish and show potential to be used as biomarkers in further toxicological evaluation studies.

Mycobacterial Interspersed Repetitive Unit-variable Number Tandem Repeat (MIRU-VNTR) Typing Lacks Discriminatory Power in the Genetic Analysis of Bovine Tuberculosis in Egypt

Tuberculosis (TB) is one of the most important infectious zoonotic diseases of vertebrates worldwide. TB in animals is primarily known from cases in cattle and other bovids for which the disease is generally referred to as bovine TB. The major causative agent of bovine TB is Mycobacterium bovis (M. bovis), a member of the Mycobacterium tuberculosis complex. Animal TB is a disease of high economic relevance within the context of livestock farming as it directly affects animal productivity and also influences international trade of animal products. In this study, we aimed at mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) analysis of cases of TB infection in local cattle in Egypt. Therefore, various samples (milk and blood samples) were collected from cattle farms (in Damietta Province) that were positive for tuberculin test. Mycobacterial isolation was tested on milk samples, but it showed negative result. DNA was extracted from blood samples. Five mycobacterial genes (IS6110, katG, gyrA, oxyR, pncA) were used for further confirmation of field TB infection. All blood samples were positive for Mycobacterial-specific genes. Twelve MIRU-VNTR loci were used to test their discriminatory power in the genetic analysis of TB, but such MIRU-VNTR loci typing failed to show any discriminatory power for the genetic analysis of bovine TB cases.

Effects of Dietary Substitution ofCod Liver Oil by Vegetable Oils onFatty Acid Composition in Liverand Muscles, Lipoprotein LipaseGene Expression and IntestinalHistomorphology in Juvenile NileTilapia (Oreochromis niloticus)

Effects of Dietary Substitution of Cod Liver Oil by Vegetable Oils on Fatty Acid Composition in Liver and Muscles, Lipoprotein Lipase Gene Expression and Intestinal Histomorphology in Juvenile Nile Tilapia (Oreochromis niloticus) This study aimed to investigate the effect of dietary substitution of cod liver oil (CLO) by vegetable oils on fatty acid composition in liver and muscles and histomorphology of proximal, mid and distal intestine in juvenile Nile tilapia (O. niloticus). Fish in duplicate were fed five isonitrogenous and isocaloric diets [cod liver oil (CLO), linseed oil (LO), olive oil (OO)] or a blend of equal proportions of CLO and vegetable oils (CLO : LO, CLO : OO) at 3 % of the diets for 60 days. The levels of lipoprotein lipase (LPL) gene expression significantly increased in the fish liver had CLO: LO diet and significantly decreased in liver of fish had OO diet.Meanwhile, nonsignificant increase of LPL geneexpression was shown in muscle of fish fed on CLO and LO diets Histomorphometric measurements made in the intestinal tract revealed that, with the exception of the luminal diameter, number of the mucosal folds and width of the lamina propria, the other histomorphometric parameters were significantly higher in the three parts in LO group than in CLO: OO and OO groups.