Biochemical and Genetical Evaluation of Pomegranate Impact on Diabetes Mellitus Induced by Alloxan in Female Rats

Abstract: Various food industries explored the possibility of developing a nutritional supplement rich in natural antioxidants from pomegranates. This study has focused on the ability of pomegranate peel and juice to study the antioxidant status. Thirty two rats were allocated in 4 groups as follows: GroupI; control group without any treatment; GroupII: diabetic animals injected with alloxan; Group III: diabetic peel group animals injected with alloxan and then feed on peel pomegranate; GroupIV: diabetic juice group animals injected with alloxan and then gavage with pomegranate juice. After 4 weeks of treatment biochemical analysis were measured such as glucose, insulin, alpha-amylase, lipid profile (cholesterol, triglyceride HDL, LDL and total lipids), total protein, homocysteine, total antioxidant capacity and liver enzymes (AST&ALT). In addition, pancreas and liver tissues were separated for genetic analysis in which pancreatic tissues were used for RAPD-PCR analysis and liver tissues for DNA fragmentation assay. Results showed significant increase in glucose and alpha amylase levels in diabetic group, while insulin decreased. Peel and juice of pomegranate ameliorates this effect and decreased glucose, alpha amylase while insulin level increased. Cholesterol, triglycerides, LDL and total lipids increased while HDL decreased in diabetic group. Peel and juice of pomegranate prevented these changes. The more pronounced effect appeared in group III treated with peel pomegranate. Total protein was not affected by alloxan or pomegranate. Homocysteine was significantly increased while total antioxidant capacity decreased in diabetic group. After treatment by pomegranate peel and juice, these parameters become near to the control values. AST and ALT were significantly increased in diabetic group. But after treatment with peel and juice, AST and ALT levels decreased and become near to the control level especially ALT value. Furthermore, rate of DNA fragmentation and DNA band polymorphism increased significantly in diabetic group. While after treatment by peel and juice rate of DNA band polymorphism and DNA fragmentation were decreased significantly. Pomegranate peel and juice showed significant reduction in LDL oxidative susceptibility and an increase in total antioxidant status. Pomegranate is able to reduce the progression in atherosclerosis. The antioxidant content in foods decreased the oxidative stress related diseases

Immunotoxicity evaluation of novel bioactive composites in male mice as promising orthopaedic implants

Objective : In orthopaedics, novel bioactive composites are largely needed to improve the synthetic achievement of the implants. In this work, semiconducting metal oxides such as SiO 2 , TiO 2 , and ZrO 2 particles (Ps) were used individually and in different ratios to obtain different biphasic composites. The immunotoxicity of these composites was tested to inspect the potential toxicity prior to their use in further medical applications. Materials and methods : In vitro mineralisation ability was inspected by soaking the composites in simulated body fluid (SBF). Additionally, in vivo experiments were performed consuming male mice using ISSR-PCR, micronucleus (MN) test, comet assay, glutathione peroxidase activity, and determination of albumin, globulin, lymphocyte population, ALT, and AST levels. Several groups of adult male albino mice were treated with 100, 200, and 400 mg/kg body weight of SiO 2 , TiO 2 , and ZrO 2 -Ps in pure or mixed forms. Results : Our findings revealed that treatment of mice with low and medium doses of SiO 2 , TiO 2 , and ZrO 2 -Ps in pure or mixed form revealed values relatively similar to the control group. However, using 400 mg/kg especially from TiO 2 -Ps in genuine form or mixed with SiO 2 showed proliferation in the toxicity rates compared with the high dose of SiO 2 and ZrO 2 -Ps. Conclusions : The results suggest that TiO 2 composite induced in vivo toxicity, oxidative DNA damage, bargain of the antioxidant enzymes, and variations in the levels of albumin, globulin, lymphocyte population, ALT, and AST in a dose-dependent manner. However, SiO 2 , and ZrO 2 composites revealed a lower toxicity in mice compared with that of TiO 2

Comparing growth, immune and pigmentation related gene expression in three lines of Japanese and wild European quail

© 2017, Polish Academy of Sciences. All rights reserved. This study was conducted to identify the differences and similarities among three Japanese quail (JQ) lines (JQ lines: white, brown and wild-black) and the European quail (EQ). The qRT-PCR was used to determine the expression of growth related genes: growth hormone (GH) and Insulin-like growth factor I (IGF-I), immune genes (Interleukins 1β, IL-1β, Interferon-α, IFN-α), pigmentation genes; dopachrome tautomerase, (Dct) and endothelin receptor type B2, (EdnrB2) in several quail tissues, while PCR-RFLP analysis of the mitochondrial 12S rRNA gene was conducted in quail meat. Expression levels of the pigmentation related genes (Dct and EdnrB2) were significantly higher (P\u3c0.05) in the JQBr and EQ lines than in JQwh and they were comparable between JQbr and wild EQ. Expression levels of the growth related genes (GH and IGF-1) were significantly higher in 3 JQ lines than in EQ. No differences between all 4 quail lines were found in the expression of the immune related genes. In conclusion, the PCR-RFLP method may be used to distinguish between the Japanese and the European quail, which is important for breeding programs, labeling meat products and biodiversity studies

Expression of Inflammatory and Cell Death Program Genes and Comet DNA Damage Assay Induced by Escherichia coli in Layer Hens.

Modern methods of industrial poultry and egg production systems involve stressful practices that stimulate Escherichia coli (E. coli) activity causing endotoxic shock. This investigation was conducted to evaluate the expression of pro-inflammatory cytokines and cell death program genes and DNA damage induced by E. coli in the brain and liver tissues of laying hens. A total of two hundred and ten H&N brown layer hens with 20 week age, were used in this research. First, preliminary experiments were designed (60 hens in total) to establish the optimal exposure dose of E. coli and to determine the nearest time of notable response to be used in the remainder studies of this research. At 35-wk of age, 150 hens were randomly assigned into 2 groups with 3 replicates of 25 birds each; the first group was injected in the brachial wing vein with 107 E. coli colony/hen, while the second group was injected with saline and served as a control. The body temperature and plasma corticosterone concentration were measured 3 hr after injection. Specimens of liver and brain were obtained from each group and the gene expression of p38 mitogen-activated protein kinase, interlukin-1β (IL-1β), tumor necrosis factor alpha (TNF-α), Bax, and caspase-3 genes were measured by quantitative real-time PCR. DNA damage in the brain and liver tissues were also measured by comet assay. Hens treated with E. coli showed significant (P<0.05) increase of body temperature and plasma corticosterone (42.6°C and 14.5 ng/ml, respectively) compared to the control group (41.1°C and 5.5 ng/ml, respectively). Additional remarkable over-inflammation gene expression of p38, IL-1β and TNF-α.genes were also detected in the brain (2.2-fold, 2.0-fold and 3.3-fold, respectively) and the liver (2.1-fold, 1.9-fold and 3.0-fold, respectively) tissues of the infected chickens. It is also important to note that hens injected with E. coli showed an increase in DNA damage in the brain and liver cells (P<0.05). These results were synchronized with activating cell death program since our data showed significant high expression of Bax gene by 2.8- and 2.7-fold and caspase-3 gene by 2.5- and 2.7-fold in the brain and liver tissues of infected chickens, respectively (P<0.05). In conclusion, the current study indicates that E. coli injection induces inflammatory physiological response and triggers cell death program in the brain and liver. Our results provide more understanding to endotoxic shock by E. coli in chickens at cellular level. Further studies are required to confirm if such responses are destructive or protective to set the means through which a chicken mounts a successful defense against avian pathogenic E. coli

Effect of <i>E</i>. <i>coli</i> on the relative expression of cell death program genes <i>Bax</i> (A) and <i>caspase-3</i> (B) in brain and liver tissues of chickens.

<p>^a,b Mean values within tissue with unlike superscript letters are significantly different (P<0.05).</p

Details of primers used for real-time PCR quantitative analysis.

<p>Details of primers used for real-time PCR quantitative analysis.</p

Expression of Inflammatory and Cell Death Program Genes and Comet DNA Damage Assay Induced by <i>Escherichia coli</i> in Layer Hens

<div><p>Modern methods of industrial poultry and egg production systems involve stressful practices that stimulate <i>Escherichia coli</i> (<i>E</i>. <i>coli</i>) activity causing endotoxic shock. This investigation was conducted to evaluate the expression of pro-inflammatory cytokines and cell death program genes and DNA damage induced by <i>E</i>. <i>coli</i> in the brain and liver tissues of laying hens. A total of two hundred and ten H&N brown layer hens with 20 week age, were used in this research. First, preliminary experiments were designed (60 hens in total) to establish the optimal exposure dose of <i>E</i>. <i>coli</i> and to determine the nearest time of notable response to be used in the remainder studies of this research. At 35-wk of age, 150 hens were randomly assigned into 2 groups with 3 replicates of 25 birds each; the first group was injected in the brachial wing vein with 10⁷ <i>E</i>. <i>coli</i> colony/hen, while the second group was injected with saline and served as a control. The body temperature and plasma corticosterone concentration were measured 3 hr after injection. Specimens of liver and brain were obtained from each group and the gene expression of <i>p38</i> mitogen-activated protein kinase, interlukin-1β (<i>IL-1β</i>), tumor necrosis factor alpha (<i>TNF-α</i>), <i>Bax</i>, and <i>caspase-3</i> genes were measured by quantitative real-time PCR. DNA damage in the brain and liver tissues were also measured by comet assay. Hens treated with <i>E</i>. <i>coli</i> showed significant (P<0.05) increase of body temperature and plasma corticosterone (42.6°C and 14.5 ng/ml, respectively) compared to the control group (41.1°C and 5.5 ng/ml, respectively). Additional remarkable over-inflammation gene expression of <i>p38</i>, <i>IL-1β</i> and <i>TNF-α</i>.genes were also detected in the brain (2.2-fold, 2.0-fold and 3.3-fold, respectively) and the liver (2.1-fold, 1.9-fold and 3.0-fold, respectively) tissues of the infected chickens. It is also important to note that hens injected with <i>E</i>. <i>coli</i> showed an increase in DNA damage in the brain and liver cells (P<0.05). These results were synchronized with activating cell death program since our data showed significant high expression of <i>Bax</i> gene by 2.8- and 2.7-fold and <i>caspase-3</i> gene by 2.5- and 2.7-fold in the brain and liver tissues of infected chickens, respectively (P<0.05). In conclusion, the current study indicates that <i>E</i>. <i>coli</i> injection induces inflammatory physiological response and triggers cell death program in the brain and liver. Our results provide more understanding to endotoxic shock by <i>E</i>. <i>coli</i> in chickens at cellular level. Further studies are required to confirm if such responses are destructive or protective to set the means through which a chicken mounts a successful defense against avian pathogenic <i>E</i>. <i>coli</i>.</p></div

Visual score of DNA damage in brain and liver tissues of chicken treated with <i>E</i>. <i>coli</i> using comet assay.

<p>Visual score of DNA damage in brain and liver tissues of chicken treated with <i>E</i>. <i>coli</i> using comet assay.</p

Effect of <i>E</i>. <i>coli</i> on the relative expression of protein kinase <i>p38</i> gene in the brain and liver tissues of chickens.

<p>^a,b Mean values within tissue with unlike superscript letters are significantly different (P<0.05).</p