Analysis of Gene Expression in Mouse Brains During Toxoplasma gondii Infection

Previous studies in our laboratory have shown that dietary supplementation with antioxidants is harmful during murine infection with the protozoan parasite, Toxoplasma gondii, which causes toxoplasmosis in humans. Dietary supplementation with vitamin E and selenium was shown to increase tissue cyst number, tissue pathology, and weight loss during T. gondii infection. The goal of the present study was to determine the impact of diet on global gene expression in the brains of infected and non-infected mice and to identify the major genes that are differentially expressed, determine their function, and discover the molecular pathways in which these genes participate. Whole genome screening, a method of determining global gene expression, was performed on RNA isolated from the brains of C57BL/6 mice, utilizing Agilent Oligo Microarrays (Agilent Technologies, Inc). Comparison of the transcripts in infected and non-infected brain tissue revealed 1,688 differentially expressed genes (p\u3c0.05) in mice maintained on the antioxidant diet. Among these genes 507 were up regulated and 71 were down regulated with 2 fold change or greater. Over 60 cytokine or cytokine-related genes showed increased expression, along with 19 up-regulated chemokine-related genes. Greater than 60% of the genes which showed elevated expression could be classified as pro-inflammatory, immune function, or cellular defense genes. In mice maintained on a diet lacking vitamin E and selenium, we observed 2,743 differentially expressed genes (p\u3c0.05) in the brains of infected mice as compared to non-infected mice. Among these genes, 239 were up regulated and 13 genes were down regulated with 2 fold or greater change. Nine chemokine or chemokine receptor genes and over 60 cytokine or cytokine receptor genes were up regulated in this comparison. The results of this study demonstrate a correlation between the increased pathology observed in infected mice maintained on an antioxidant-supplemented diet and an intense and extensive pro-inflammatory response in brain tissue

Reactive Oxygen Species

The term “reactive oxygen species” (ROS) refers to a group of reactive molecules and free radicals produced by molecular oxygen. In recent decades, there has been great interest in the role of ROS in various diseases. From basic science research to clinical trials, biomedical scientists have made rapid progress toward a better understanding of ROS-metabolizing systems and their role in health and diseases. This book includes sixteen chapters that address topics such as the history of ROS, its role in autoimmunity, neurodegeneration, and aging, and recent advances in various antioxidants and their therapeutic potential

EFFECTS OF MELON PULP RICH IN SUPEROXIDE DISMUTASE ON PIGLETS AND POULTRY HEALTH

The aim of the present thesis was to assess the effect of the dietary supplementation of SOD-rich melon pulp concentrate (MPC) on oxidative status, inflammation and growth performance of post-weaning piglets and poultry. With this purpose three different trials were conducted: 1) MPC administration to no challenge or LPS-challenged post-weaning piglets to evaluate changes in the oxidative status and inflammation markers in blood together with growth performance; 2) MPC administration to broilers to evaluate growth performance and the incidence of both pododermatitis and cellulitis; 3) expression of oxidative status genes in the liver in both piglets and chickens. During trial 1, forty-eight female piglets weaned at 24 days of age were divided in four homogeneous experimental groups of twelve animals each in a randomized block design and fed a basal diet (C, n. 24) or a basal diet plus 30 g/ton melon pulp concentrate (Melofeed\uae, Lallemand, Blagnac, France) (MPC, n. 24). The experimental trial lasted 29 days from weaning, performing a LPS challenge by injecting increased dosages four times every two days starting from 19th day of the experiment (19, 21, 23, 25 days) in half of C (n.12) and half of MPC (n.12) groups. Initial LPS dosage of 60 \u3bcg/kg of body weight was increased by 12% at each subsequent injection to reduce endotoxin tolerance. Growth performance including live body weight, feed intake, average daily gain and G:F were monitored from the beginning of the trial until the end of the experiment. Blood samples were collected to evaluate the antioxidant status of experimental animals by SOD activity (Superoxide dismutase), TAOC (Total Anti-Oxidant Capacity), ROS (Reactive Oxygen Species), Kit Radicaux Libres (KRL) test on blood, plasma and red blood cell (RBC), and 8-oxodGuo (8-oxo-7, 8-dihydro-2\u2019-deoxyguanosine). Moreover blood samples were analysed for immune and inflammatory response by haptoglobin (Hp) and Cytokines IL-6, IL-1\u3b2, TNF\u3b1 in serum. A positive effect of SOD-rich MPC was evidenced on FI and ADG (P<0.01; P=0.05 respectively), while LPS challenge significantly reduced both parameters in injected animals (P<0.01). Antioxidant status of MPC fed piglets was improved by higher TAOC levels (MPC=7.22 mM Trolox equivalent vs. C=4.54 mM Trolox equivalent; P<0.01) and RBC resistance to haemolysis (MPC=70.71 HT50, min vs. C=66.41 HT50, min; P 640.01). No significant differences were evidenced for ROS, SOD activity and 8-oxodGuo in the four experimental groups. The challenge with LPS increased TNF-\u3b1, IL-1\u3b2, IL-6 (P<0.01) and Hp (P=0.03) levels in blood, but no differences were found for MPC administration. These results suggest that oral SOD supplementation by MPC increase some aspects of antioxidant status of post-weaning piglets with positive results on growing performance. In trial 2 a total of 1104 broilers were allocated to 4 experimental groups for 35 days with four different dietary treatments: a) basal diet (C: control), b) basal diet plus melon pulp concentrate (MPC1, MPC2 and MPC3) as described below. The feeding regimen consisted of starter (0-10 d), grower (11-24 d) and finisher diet (25-35 d). During starter phase, dietary treatments were corn-soybean meal based diets supplemented with 0 g/ton (C), 30 g/ton (MPC1), 15 g/ton (MPC2), 15 g/ton (MPC3) of melon pulp concentrate (MPC) (Melofeed\uae, Lallemand, Blagnac, France). In grower phase, C, MPC1 and MPC2 received same basal diet but MPC3 received basal diet supplemented with 15 g/ton of MPC. During 24-35 days, the same basal diet without MPC was supplied to four experimental groups. 12 pens per group with 23 broilers per pen were used. On day 24 and 25, pododermatitis was evaluated on all animals using a scoring system that ranged from 0 to 2. On day 35, cellulitis was evaluated on all slaughtered birds. Litter score and litter DM was assessed on day 35. Body weight (BW) of birds per pen was recorded on 0, 10, 24, 35 experimental days from the beginning of the experiment. Individual body weight, slaughter live body weight, carcass weight, dressing percentage and organs weight (liver, heart, intestine, Bursa of Fabricius, spleen and pancreas and gizzard) percentage were also assessed at day 35. Pen feed intake was recorded on 0, 10, 24, 35 experimental days. Pen feed residues were determined at the end of each feeding period to estimate mean ADG, ADFI and gain: feed (G:F) ratio for each pen. Treatment (MPC) was able to affect body weight (P<0.05), ADG (P<0.05) and G:F (P=0.05) during experimental period. Final BW was higher in MPC1 and MPC2 than C and MPC3 (P<0.01). Carcass weight tended to be higher in MPC2 than C and MPC3 (0.05<P 640.1). Percentage of the Bursa of Fabricius weight tended to be higher (0.05<P 640.1) in MPC1 and MPC3. Incidence and severity of pododermatitis varied within each feeding period. Significant treatment, time and their interaction (P<0.01) were found in pododermatitis lesions analysis. At 24 d of age, MPC3 scores were the highest (P<0.01) between diets. At 35 d of age, scores in MPC3 were still higher than MPC1 (P<0.05). Pearson correlation between pododermatitis and body weight was not significant, by the way it was highly significant (P<0.01) between pododermatitis and litter dry matter (-30%). No cellulitis was detected in experimental animals. These results suggest that oral SOD supplementation by MPC increases performance of poultry with a dose-dependent increase in pododermatitis incidence, but not significant effect on cellulitis occurrence. In the third trial liver tissue was collected from six piglets per treatment at the end of the trial 1 and 12 poultry per treatment at the end of grower phase (day 24) from trial 2 respectively. Collected samples were stored in liquid nitrogen for subsequent gene expression analysis by RT-PCR. \u3b2-actin and GAPDH of S. scrofa or G. gallus respectively were used as internal reference genes. GPx1, CAT and SOD1 were determined on piglet liver samples, while NFEL2L, CAT and SOD1 were determined on poultry hepatic tissue samples. Results from piglets\u2019 trial showed that MPC, LPS challenge and MPC x LPS challenge interaction did not significantly affect liver gene expression of GPX1, CAT and SOD1 (P>0.05), with the exception of trend (P=0.09) for a down-regulation of CAT for MPC effect. Results from poultry trial showed that MPC did not significantly affect liver gene expression of NFEL2L in treated groups, while SOD1 expression showed a trend to reduce the down-regulation in MPC3 vs. C (P=0.09). CAT was significantly up-regulated in MPC3 than C and MPC1 (P=0.02). The obtained results show that MPC supplementation can directly affect CAT expression in the liver of poultry and challenged piglets and, to a lesser extent SOD1 in poultry. Results from the present thesis show that the inclusion of SOD-rich melon pulp concentrate in the diet of post-weaning piglets and poultry can significantly increase the blood levels of some antioxidant biomarkers and the expression of genes implicated in the regulation of the oxidative status in the liver

Vitamin C in Health and Disease

Vitamin C is a pivotal water soluble electron donor in nature and an essential nutrient in man. Despite its many years as a research focus, new and increasingly regulatory functions of vitamin C in human health are continually being unravelled. This improved mechanistic insight is starting to provide rationales explaining the extensive epidemiological literature that, for decades, has consistently shown strong associations between poor vitamin C status and increased morbidity and mortality. In this Special Issue, we include original research and literature reviews by experts in the field outlining the roles of vitamin C in early, daily and late life, as well as the roles of deficiency in cardiovascular disease, inflammation and cancer

Viral infections in 47 CVID patients in allergy and immunology department of Rasool E Akram hospital in Tehran

Background: CVID is a heterogeneous primary immune deficiency with infectious, autoimmune and autoinflamatory features. It is most common symptomatic PID in Iran, with prevalence of 1 in 25000 to 50000 people. CVID has been divided into some phenotypes to produce more homogenized subpopulations. CVID is not a pure Ab deficiency .and because of both abnormalities in Tcell and innate immunity in combination with B cell dysfunction these patients are predisposed to viral and opportunistic infections. Method: prevalence of viral infections is reported in 47 CVID patients registered in Rasool E Akram hospital in Tehran. Patients have been diagnosed as CVID with the PAGID-ESID diagnostic criteria in our department or referred from other clinics for follow up and treatment. Diagnosis of viral germs has been made by clinical signs, pathological significances and in some cases by PCR. Cases: 9 patients (19%) had problems with viral infections. Infections occurred befor diagnosis of CVID in some cases or after that. Four patients (8.5 %) had problems with wart. Sever mucocutaneus HSV infection has occurred in 3 (6 %), recurrent zona in one (2 %) and CMV infection as colitis or pneumonitis in 3(6 %) patients. Sever progressive lethal CNS infection with JC virus occurred in one patient. Conclusion: evidences show that CVID is not a pure B cell defect, and we should be aware of opportunistic and viral infections that in some cases may be fatal

The Practice of Sheep Veterinary Medicine

This book is intended to be a reference text for veterinarians who provide clinical services to sheep producers. It is directed first and foremost at Australian sheep-raising systems, but the approaches described herein will have wide application in all countries where sheep are raised under extensive grazing conditions. Most of the important conditions of sheep in Australia are relatively straightforward to diagnose, but the establishment of effective and economically sound control strategies is often the most difficult part of health management, particularly for those who are less familiar with sheep production systems. With six initial chapters focusing on providing readers with a basic understanding of the business and science underpinning sheep production, this book focuses its remaining chapters on reproduction and disease conditions, ordered largely on a systems basis. The book provides details about the way disease processes develop and manifest in sheep flocks, with numerous references for those who wish to read further. Australian sheep production is a profitable and fulfilling agricultural pursuit for a large number of farm owners, and this book is intended to assist those who work in the industry to add to the profitability and efficiency of sheep production systems, the quality of sheep products and the welfare of the sheep in those systems

Current Perspectives in HIV Infection

This book gives a comprehensive overview of HIV and AIDS including NeuroAIDS, as well as general concepts of pathology, immunity and immunopathology, diagnosis, treatment, epidemiology and etiology to current clinical recommendations in management of HIV/AIDS including NeuroAIDS, highlighting the ongoing issues, recent advances and future directions in diagnostic approaches and therapeutic strategies

Chromatin and Epigenetics

Genomics has gathered broad public attention since Lamarck put forward his top-down hypothesis of 'motivated change' in 1809 in his famous book "Philosophie Zoologique" and even more so since Darwin published his famous bottom-up theory of natural selection in "The Origin of Species" in 1859. The public awareness culminated in the much anticipated race to decipher the sequence of the human genome in 2002. Over all those years, it has become apparent that genomic DNA is compacted into chromatin with a dedicated 3D higher-order organization and dynamics, and that on each structural level epigenetic modifications exist. The book "Chromatin and Epigenetics" addresses current issues in the fields of epigenetics and chromatin ranging from more theoretical overviews in the first four chapters to much more detailed methodologies and insights into diagnostics and treatments in the following chapters. The chapters illustrate in their depth and breadth that genetic information is stored on all structural and dynamical levels within the nucleus with corresponding modifications of functional relevance. Thus, only an integrative systems approach allows to understand, treat, and manipulate the holistic interplay of genotype and phenotype creating functional genomes. The book chapters therefore contribute to this general perspective, not only opening opportunities for a true universal view on genetic information but also being key for a general understanding of genomes, their function, as well as life and evolution in general