From olfaction to immunity: Characterization of nasal immunity in bony fish

The olfactory system is a common route pathogen entry in vertebrates. As a consequence, the nasopharynx-associated lymphoid tissue (NALT) needs to rapidly clear infections without compromising the sense of olfaction. NALT is present in teleost fish but its cellular and molecular mechanisms of action have not been investigated to this date. This dissertation focuses on three aims: 1. investigating the role of CCL19-like as a primordial chemokine in vertebrate nasal immunity, 2. determining the presence of tissue microenvironments within the olfactory organ (OO) of rainbow trout, and 3. understanding the immune contributions of olfactory sensory neurons (OSNs) in teleosts against viruses. In aim 1, we report six isoforms of CCL19-like chemokine in salmonids such as CK12. CK12 is mainly expressed in mucosal tissues and plays an important role in antiviral immunity. Although recombinant protein CK12 is not chemotactic in vitro, it induces infiltration of APCs and CD8+ T cells into OO of rainbow trout in vivo. In aim 2, we demonstrate the presence of two different microenvironments (mucosal tip and neuroepithelial) in the OO of rainbow trout. The tip of the epithelium harbors clusters of CD8α+ cells whereas few numbers are found in the neuroepithelium. CD8a+ cell localization corresponds with a higher expression of chemokine and chemokine receptors in the tip versus the neuroepithelial side. We report that cell proliferation in response to viral nasal delivery occurs mainly at the tip. In aim 3, we unravel for the first time a new function for OSNs by which nasal rhadboviruses induce apoptosis in crypt neurons, a type of OSN, in rainbow trout via the interaction of viral G glycoprotein and the neuron TrkA receptor. CD8α+ T cells infiltrate to the OO within minutes of nasal viral delivery and this response was abrogated when TrkA was blocked. Infiltrating CD8α+ T cells originated from the microvasculature surrounding the olfactory bulb (OB) and not the periphery. In conclusion, this dissertation provides the first tissue, cellular and molecular characterization of teleost NALT and reveals a novel function of vertebrate OSNs in eliciting rapid nasal anti-viral immune responses in the OO and OB

Indolepropionic acid and novel lipid metabolites are associated with a lower risk of type 2 diabetes in the Finnish Diabetes Prevention Study

Wide-scale profiling technologies including metabolomics broaden the possibility of novel discoveries related to the pathogenesis of type 2 diabetes (T2D). By applying non-targeted metabolomics approach, we investigated here whether serum metabolite profile predicts T2D in a well-characterized study population with impaired glucose tolerance by examining two groups of individuals who took part in the Finnish Diabetes Prevention Study (DPS); those who either early developed T2D (n = 96) or did not convert to T2D within the 15-year follow-up (n = 104). Several novel metabolites were associated with lower likelihood of developing T2D, including indole and lipid related metabolites. Higher indolepropionic acid was associated with reduced likelihood of T2D in the DPS. Interestingly, in those who remained free of T2D, indolepropionic acid and various lipid species were associated with better insulin secretion and sensitivity, respectively. Furthermore, these metabolites were negatively correlated with low-grade inflammation. We replicated the association between indolepropionic acid and T2D risk in one Finnish and one Swedish population. We suggest that indolepropionic acid, a gut microbiota-produced metabolite, is a potential biomarker for the development of T2D that may mediate its protective effect by preservation of alpha-cell function. Novel lipid metabolites associated with T2D may exert their effects partly through enhancing insulin sensitivity.Peer reviewe

Regimen adherence and self-management in persons with impaired glucose tolerance and Type 2 Diabetes

With the rising trend of obesity within the United States, the prevalence of Type 2 Diabetes (T2D) is increasing as well. The best way to stop the growing numbers of T2D is to prevent it completely. However, for those who already have T2D or Impaired Glucose Tolerance (IGT) healthy management of the disease is crucial for lowering life threatening complications and lowering mortality. There is strong evidence supported by decades of research that a lifestyle intervention (increased physical activity and healthy eating habits) is the best way to prevent T2D and to manage the disease. This type of management structure leaves out the humanity of the disease and doesn't account for the psychosocial factors at play. Depression, social support, self-care, coping mechanisms, and education on nutrition are a few of the psychosocial factors that are heavily evaluated in this study. The purpose of the present study is to calculate the magnitude of effect of a lifestyle intervention with a social support group on self-management and regimen adherence, HbA1c, body weight, and body fat percentage. This study included a 12-week lifestyle intervention with the intervention group partaking in a weekly support group. All participants were given weekly educational materials covering a biopsychosocial spectrum. In addition to the biomarkers mentioned, 6 survey instruments were used to measure self-management and regimen adherence from baseline to end of study. Following the 12-weeks, the mean increase in minutes walked weekly was 197.5 min. Analysis on the survey data showed the intervention group lowered their diabetes related distress, increased self-care habits, and decreased utilization of food as a coping mechanism. The control group data showed a higher level of diabetes related distress and a lower level of self-control from baseline to end of study. Both groups showed a decrease in self-efficacy and self-control from baseline to end of study. These findings are meaningful and add to the small amount of research already done on biopsychosocial lifestyle interventions for persons with T2D and IGT. Larger scale studies will need to be done in the future to increase the validity of statistical findings

DECONTAMINATION SYSTEMS AND INFORMATION RESEARCH PROGRAM

During the five plus years this Cooperative Agreement existed, more than 45 different projects were funded. Most projects were funded for a one year period but there were some, deemed of such quality and importance, funded for multiple years. Approximately 22 external agencies, businesses, and other entities have cooperated with or been funded through the WVU Cooperative Agreement over the five plus years. These external entities received 33% of the funding by this Agreement. The scope of this Agreement encompassed all forms of hazardous waste remediation including radioactive, organic, and inorganic contaminants. All matrices were of interest; generally soil, water, and contaminated structures. Economic, health, and regulatory aspects of technologies were also within the scope of the agreement. The highest priority was given to small businesses funded by the Federal Energy Technology Center (FETC) and Department of Energy (DOE) involved in research and development of innovative remediation processes. These projects were to assist in the removal of barriers to development and commercialization of these new technologies. Studies of existing, underdeveloped technologies, were preferred to fundamental research into remediation technologies. Sound development of completely new technologies was preferred to minor improvements in existing methods. Solid technological improvements in existing technologies or significant cost reduction through innovative redesign were the preferred projects. Development, evaluation, and bench scale testing projects were preferred for the WVU research component. In the effort to fill gaps in current remediation technologies, the worth of the WVU Cooperative Agreement was proven. Two great technologies came out of the program. The Prefabricated Vertical Drain Technology for enhancing soil flushing was developed over the 6-year period and is presently being demonstrated on a 0.10 acre Trichloroethylene contaminated site in Ohio. The SpinTek Centrifugal Membrane System was a unique separation process introduced through the Agreement that is now being used at the Los Alamos National Laboratory. Based on the cost to the USDOE for both technologies and considering their usefulness in cleaning up contaminated sites, no other technologies developed through USDOE provide or have the propensity to provide as great a return on investment and impact on environmental remediation. These technologies alone make the $10.3 million USDOE investment in the WVU Cooperative Agreement a tremendous investment

Changing expression of vertebrate immunity genes in an anthropogenic environment: a controlled experiment

Background: The effect of anthropogenic environments on the function of the vertebrate immune system is a problem of general importance. For example, it relates to the increasing rates of immunologically-based disease in modern human populations and to the desirability of identifying optimal immune function in domesticated animals. Despite this importance, our present understanding is compromised by a deficit of experimental studies that make adequately matched comparisons between wild and captive vertebrates. Results: We transferred post-larval fishes (three-spined sticklebacks), collected in the wild, to an anthropogenic (captive) environment. We then monitored, over 11 months, how the systemic expression of immunity genes changed in comparison to cohort-matched wild individuals in the originator population (total n = 299). We found that a range of innate (lyz, defbl2, il1r-like, tbk1)and adaptive (cd8a, igmh) immunity genes were up-regulated in captivity, accompanied by an increase in expression of the antioxidant enzyme, gpx4a. For some genes previously known to show seasonality in the wild, this appeared to be reduced in captive fishes. Captive fishes tended to express immunity genes, including igzh, foxp3b, lyz, defbl2, and il1r-like, more variably. Furthermore, although gene co-expression patterns (analyzed through gene-by-gene correlations and mutual information theory based networks) shared common structure in wild and captive fishes, there was also significant divergence. For one gene in particular, defbl2, high expression was associated with adverse health outcomes in captive fishes. Conclusion: Taken together, these results demonstrate widespread regulatory changes in the immune system in captive populations, and that the expression of immunity genes is more constrained in the wild. An increase in constitutive systemic immune activity, such as we observed here, may alter the risk of immunopathology and contribute to variance in health in vertebrate populations exposed to anthropogenic environments

Survey of hydrogen production and utilization methods. Volume 2: Discussion

For abstract, see N76-15590

Non-specific immune response and disease-resistance in Chinook salmon (Oncorhynchus tshawytscha) supplemented with probiotics

Finfish aquaculture is the fastest-growing food-production sector in the world. Nevertheless, intensive production faces increased disease outbreaks due to high stock densities and environmental effects. Currently, infectious diseases represent the main causes of losses in salmon aquaculture. To prevent this, antibiotics and vaccines have been frequently utilized as a treatment and control methods for infectious agents. However, restrictions on antibiotic use due to antibiotic-resistant strain outbreaks, as well as, the variable protection exerted by vaccines, has made the industry to explore additional treatments. Probiotics are an important non-toxic and non-polluting tool in aquaculture to improve fish growth, stress tolerance and non-specific defense. In finfish aquaculture, many probiotics have been tested, including lactic acid bacteria (LAB; Lactobacillus and Carnobacterium), Bacillus, and Pediococcus spp. The role of probiotics within the digestive tract in the two most important salmonids, Atlantic salmon and rainbow trout, has been widely studied, however the effect of probiotic supplementation on the growth, survival, immune response, and gut integrity of Chinook salmon (Oncorhynchus tshawytscha) raised under aquaculture conditions has not been widely studied. Chinook salmon is a native species of the Canadian Pacific coast with potential economic and environmental benefits for aquaculture. Additionally, Chinook salmon farming can play an important role in diversifying current Canadian aquaculture and help protecting the wild stocks. Some issues that keep to date Chinook salmon farming at a low scale are the risk of escapees diluting the genetic diversity of wild populations and the decrease of the flesh quality when they reach sexual maturation. Sterile triploid salmon offer a solution however, they have an a 10-30% higher disease mortality rate compared to diploid fish. This makes Chinook salmon an ideal candidate to utilize probiotics as an alternative to antibiotics and vaccines. The contents of this thesis examine the effect of single-strain and multi-strain probiotics either commercially available or generously donated by companies. Also, this thesis focuses in evaluating probiotic supplementation at different production cycles (freshwater and saltwater), fish size (juveniles and adults), and/or supplementation time (4, 10, 14 months). Since in vivo experiments were conducted once a year, preliminary studies to determine the safety of the strains were carried out in the lab using the salmonid intestinal epithelial cell line derived from rainbow trout (Oncorhynchus mykiss), RTgutGC. Overall, the results obtained in this thesis demonstrated that probiotic strains utilized in different experiments did not show negative effects in terms of cell viability and/or Chinook salmon tissues (histopathological analysis). Data collected during the in vitro trials demonstrated that probiotic stimulation was able to modulate the expression of TJ and immune genes without impacting the cell viability. For the in vivo trial, the most interesting results associated to fish survival was observed after multi-strain Jamieson® probiotic supplementation throughout the freshwater and saltwater production cycle (14 months of supplementation), showing a decrease of ~10% in the mortality of Chinook salmon challenged with V. anguillarum compared to fish fed regular pellet. In all the trials conducted for this thesis, it was not possible to obtain an improvement in fish growth parameters (weight and length), as has been observed in previous probiotic supplementation studies. Nevertheless, research conducted for this thesis represents one of few studies that has been able to evaluate probiotic supplementation over a full year of saltwater production in real aquaculture conditions. The gene expression results suggest that probiotic supplementation can differentially modulate the early inflammatory response and gut barrier function after infection with V. anguillarum, however more studies are still needed to fully understand the correlation between gene expression and the survival and growth data collected in this study. The resulting data collected from the ELISAs of IgM and IgT provided novel functional information regarding protein levels in different tissues of Chinook salmon. It is important to mention that this study represents the first time that IgT of Chinook salmon has been observed in western blots and the protein levels measured in ELISA, therefore, this thesis would be the primary reference for any future studies evaluating this immunoglobulin in the future. Overall, these studies demonstrate the importance of investigating the mechanism underlying host-microbe interactions to help improve Chinook salmon aquaculture in an environmentally sustainable manner

Awareness of Diabetes Risk and Adoption of Diabetes Risk Reduction Behaviors in the Presence of Other Risk Factors in U.S Adults: An Examination of NHANES Data 2007-2008

ABSTRACT Background: Prediabetes is a precursor condition to type 2 diabetes mellitus. Previous research and clinical trials have shown that the onset of type 2 diabetes could be delayed or prevented through structured life style modifications such as dietary changes, modest weight loss and moderate-intensity exercise. This study examines U.S adults of different ethnicities that include non-Hispanic white, non-Hispanic black and Mexican Americans and whether their awareness of diabetes risk is associated with their participation in diabetes risk reduction behavior, a combination of physical activity, weight control and fat/calories intake. Methods: The 2007-2008 National Health and Nutrition Examination Survey, NHANES, was used to conduct a cross-sectional study of 4083 U.S. adults who were 20 years old and above and were aware of their diabetes risk. The association between the awareness of one’s diabetes risk and the adoption of diabetes risk reduction behavior were examined in present of other risk factors such as age, gender, ethnicity, education, annual family income, BMI, hypertension, mean systolic blood pressure, mean diastolic blood pressure, total cholesterol, LDL, HDL and triglyceride levels. Males and females were examined separately for all analyses performed. Cross tabulation was conducted and p-values were calculated by the Pearson’s chi-square test for the categorical variables which include gender, ethnicity, education, annual family income, adiposity and hypertension. One Way ANOVA and Tukey post hoc tests were conducted for the continuous variables which include age, mean systolic blood pressure, mean diastolic blood pressure, total cholesterol, LDL, HDL and triglyceride levels. Univariate and multivariate analyses were performed to determine the association between the main independent variable, awareness of one’s diabetes risk, and the dependent variable, adoption of diabetes risk reduction behavior, controlling for other risk factors. A p-value of \u3c0.05 and 95% confidence intervals were used to determine statistical significance throughout all analyses performed. Results: After controlling for age, gender, race, education, annual family income, BMI, hypertension, mean systolic blood pressure, mean diastolic blood pressure, total cholesterol, LDL, HDL, and triglycerides, results from the multivariate analysis showed that subjects who were aware of their diabetes risk were more likely to adopt diabetes risk reduction behavior (OR= 1,734, 95 % CI=1.217-2.470). Females and non-Hispanic blacks, who were aware of their diabetes risk, were also more likely to adopt diabetes risk reduction behavior compared to males, non-Hispanic whites and Mexican Americans. An increase in the levels of education, annual family income and BMI was also associated with the adoption of diabetes risk reduction behavior. Stratification according to gender and ethnicity, showed that Mexican American males and females were more likely to engage in diabetes risk reduction behavior compared to non-Hispanic whites and non-Hispanic blacks (Mexican American males: OR: 2.496, CI: 0.792-7.868; Mexican American females: OR: 2.830, CI: 0.917-8.736). Conclusion: This study provides useful insights for health care providers and public health professionals who are developing health promotion and prevention interventions to address pre diabetes before it develops into type 2 diabetes. This study also allows the development of tailored interventions for specific genders and ethnic groups that are at risk. Results of this study indicate that Mexican Americans and females (in general) are more likely to adopt diabetes risk reduction behavior. Therefore, physicians and health care providers should develop culturally, linguistically and gender- specific education materials and programs for this particular gender and ethnic group. This in turn, may reduce the overall increasing prevalence of diabetes, reduce racial and gender disparities and may have a positive impact on the overall health of the U.S. population