There are over 15.5 million cancer survivors in the U.S. currently, increasing to over 20 million by 2026. Long-term cancer survival has raised awareness for the issues that affect quality of life (QoL) after treatment. Fatigue and subjective cognitive dysfunction are common quality of life concerns for survivors but little is known regarding prevalence of these problems in daily life. The purpose of the current project is to examine these concerns after treatment using data from a 2 week daily diary study of breast cancer survivors up to 3 years post-treatment. Of importance is determining the factors that contribute to reporting decreased QoL. Taken together, our findings suggest that within-persons, survivors who reported worse mood and physical function tended to also report higher levels of fatigue. Similarly, subjective reporting of cognitive function is influenced by current mood and physical symptoms such as fatigue. Demographic characteristics and depressive symptoms prior to the study were unrelated to fatigue and subjective cognitive function. This dissertation advances the current understanding of daily QoL issues. These findings also highlight the importance of capturing these experiences in daily life.

This study examines the role of psycho-social factors in contributing to chronic illness, in particular Coronary Heart Disease (CHD) and cancer. Research conducted over the last 50 years indicates a modest role for psycho-social factors as risk factors for these conditions. The research suggests that in combination with other modest risk factors, psycho-social factors play an equally important role. Grossarth-Maticek & Eysenck (1990) gained notoriety by reporting research results which seriously question current wisdom. Grossarth-Maticek & Eysenck (1990) conducted what must be considered the largest long term prospective psychological health study. They followed around 20,000 probands for 15 years in order to determine the relative contributions to health outcomes (primarily cancer and CHD) of standard medical and psycho-social risk factors. They reported an 81% accuracy of prediction of death by cancer or CHD with the use of a psychometric test. Further, answers to the test were reported to be six times more predictive of cancer or CHD than were any of the standard medical risk factors, either taken on their own or together. Being far in excess of claims made by any other researchers, the treatment/prevention implications of Grossarth-Maticek’s study are far reaching. It is an important endeavor to examine possible influencing factors in these results. While scientific fraud can never be excluded, in the absence of any incriminating evidence, this assertion is considered to be non-empirical. Amelang (1991) wonders whether unknown, favourable and non replicable conditions were present in Grossarth-Maticek’s samples or methodology which influenced the results. The current research program represents an attempt to ascertain what some of those factors may have been. It is suggested here that Grossarth-Maticek’s probands learnt of their cancer or CHD proneness in the course of the interview which formed the basis of the data collected in their study. Focusing on this aspect of Grossarth-Maticek’s data collection methodology, the possibility of a large scale treatment effect emerges. The most psychologically vulnerable of his probands, having been stimulated by the interview process, may have ruminated and ‘catastrophized’ over their health prospects during the ensuing years, thereby making themselves physiologically more vulnerable to the feared conditions. The possibility of probands being adversely influenced by the interview is tested in the current study with subjects being measured for changes in cancer/CHD risk perception and anxiety levels as a result of participation in the ‘Grossarth-Maticek interview’ format. In order to test this hypothesis, two studies were undertaken in which Grossarth-Maticek’s data collection method was replicated with 105 subjects. Subjects were assessed on a range of self report items prior to the ‘Grossarth-Maticek’ interview, and again post to this interview. They were assessed as to their beliefs about vulnerability to cancer and CHD; and their levels of anxiety in relation to contracting these conditions. Subject’s responses to learning of their ‘health prediction’ according to Grossarth-Maticek’s assessment tool were also measured via a heart rate monitor. Changes in subject’s self report measures and heart rate responses were noted and associations to the type of health prediction which they were given at the end of the interview was assessed. In the second study, the relative effects of family history, Neuroticism and time on disease risk perception and anxiety were studied via statistical analyses of self report measures. The results of this study do not support the notion of the interview having any adverse influence. Problems with the methodology and sample selection may have influenced the outcome. It is concluded that for these reasons, the current study did not represent a thorough test of the current hypothesis. Recommendations for a superior test of this hypothesis are made.

Herpes simplex viruses (HSV) infection is a common cause of ocular disease and can result in a chronic inflammatory reaction that impairs vision. This latter manifestation is called herpetic stromal keratitis (HSK) and usually occurs as a consequence of virus reactivation from latency in the trigeminal ganglion. The pathogenesis process involves complex interactions of cellular and molecular events. HSK in humans, and certainly its murine experimental model, appears to be an immunopathologic disease process. The crucial cell type orchestrating the inflammation is a CD4+ Tcell that has a Thl cytokineproducing profile. However, prior to this immunoinflammatory phase, multiple events occur that set the stage for subsequent pathology. These include production of cytokines and chemokines, infiltration of innate immune cells and neovascularization of the avascular cornea. Current understanding about human and murine HSK pathogenesis is reviewed in Part I. Part II progresses this knowledge using knockout and transgenic mice. Results in Part II clarify the essential role of IL-l, a proinflammtory cytokine, produced as a consequence of virus replication, in HSK pathogenesis. Part III, in addition to Part II, elucidate the mechanisms involved in IL-l and IL-6 mediated polymorphonuclear leukocyte (PMN) influx and neovascularization of cornea following virus infection. These cytokines were shown to be produced quite early after infection and capable of inducing angiogenic factors, resulting in angiogenesis of the cornea. In addition, the role of IL-l in inducing proinflammatory arachidonic acid metabolites has been discussed in PART IV. Cycloxygenase 2 enzyme produced from resident corneal cells in presence of IL-l was shown to be crucial for creating an inflammatory milieu and facilitate early PMN influx in murine cornea. These results provide novel insights into the link between viral infections, pro-inflammatory mediators, PMN migration, angiogenesis and HSK development. Our understanding about the role of IL-l in HSK pathogenesis was evaluated clinically in Part V using an IL-l receptor antagonist (IL-l ra) protein. Mice receiving IL-l ra h~d diminished disease severity. The administration of IL-l ra was shown to reduce the influx into the cornea of cells of both the innate and adaptive immune response. In addition, the treatment also diminished corneal VEGF levels resulting reduced angiogenic response. Our results show the iluportance of targeting early proinflammatory molecules such as IL-l to counteract HSK and advocate IL-l ra as an effective agent to achieve this. In Part VI, a novel flow cytometry based assay has been used to quantify corneal angiogenesis following ocular HSV infection. Our results indicate that, this assay was sensitive enough to able to pick up the difference in angiogenic response between mice. Thus, estimation of angiogenesis on the basis of number of endothelial cells proved to be a useful approach to quantify corneal neovascularization. This dissertation presents research aimed at elucidating both molecular and cellular events in HSK pathogenesis. These results will serve as guidelines for future development of more efficient prophylactic and therapeutic strategies.

Primary and higher order RNA structures in the 5’ and 3’ untranslated regions of the genomes of positive-strand RNA viruses are known to function as cis-acting elements for genome translation and replication. Four cis-acting replication elements have been identified in the bovine coronavirus (BCoV) 3’ untranslated region that are postulated to recruit viral and cellular proteins for assembly of the replication complex and initiation of negative-strand synthesis. This study had the goal of identifying and establishing the function of the viral proteins that bind BCoV 3’ terminal cis-acting RNA structures. A major surprising discovery was a specific interaction of the BCoV-encoded 2’-O-methyltransferase (2’-O-MT), an enzyme normally involved in 5’-terminal cap methylation, with a 3’-terminal cis-acting octamerassociated bulged stem-loop. This discovery led to detailed analyses of BCoV 2’-O-MT that is summarized as follows: (i) Enzyme probing revealed the higher-order structure of the cis-acting 3’-terminal bulged stem loop. (ii) Specific binding of the 2’-O-MT to both the 5’ cap-0 structure and to the 3’-terminal bulged stem-loop was documented. (iii) Enhanced translation of a viral genomic RNA was demonstrated as a function of 2’-O-MT methylating activity. (iv) A requirement for 2’-O-MT methylating activity for virus replication was demonstrated with the use of the mouse hepatitis coronavirus reverse genetics system. (v) Over-expression of the viral 2’-O-MT in virus-infected cells had no detrimental in-trans effect on viral genome replication, transcription, or translation, but affected DI RNA accumulation in-cis. These results indicate that the coronaviral 2’-O-MT is essential for virus genome translation and replication and, as a multifunctional protein, is an attractive target for development of an anticoronaviral therapeutic drug.

Adipose tissue secretes over 100 different proteins and cytokines called adipokines. Adiponectin is one of the most intriguing adipokines because it is closely associated with insulin sensitivity and is an early marker for Type-2 diabetes mellitus in human beings. Cats are at risk for developing Type-2 diabetes with obesity, so it is important for researchers to understand the role adipokines, like adiponectin, play in feline metabolism. This project sought to lay the foundation for future research regarding feline adiponectin by sequencing adiponectin cDNA, measuring adiponectin’s expression in various tissues, validating and developing new techniques for measuring adiponectin in circulation, determining the influence of gender on adiponectin concentrations, and by monitoring changes in adiponectin as cats gain and lose fat mass. The results of this project revealed that adiponectin has a similar cDNA and amino acid sequence to other species and is secreted almost exclusively from adipose tissue. However, cats differed from other species in that visceral adipose tissue had more adiponectin expression than subcutaneous adipose tissue. Serum feline adiponectin was measured using a commercially available murine adiponectin ELISA, and the high molecular weight (HMW) form was detected using gel chromatography combined with ELISA. Neutered male cats may have lower concentrations of total adiponectin than female cats and lower percentages of HMW adiponectin than all other gender groups. Similar to humans, HMW adiponectin was more closely associated with improved glucose metabolism than total adiponectin. HMW adiponectin also correlated more closely with body fat mass than total adiponectin. Despite some unique characteristics, feline adiponectin appears similar in structure and function to adiponectin in other species.

Matrix metalloproteinases (MMP’s) and cathepsins are thought to play key roles in oral squamous cell carcinoma (OSCC) invasion and metastasis. Three OSCC human cell lines, BHY, HN and HSC-3, have been studied based on their reported ability to invade adjacent bone or metastasize to lymph nodes and/or distant organs. The working hypothesis of this study is that OSCC that demonstrate different actions have different proteolytic enzyme and gene expression profiles. Immunocytochemistry and flow cytometry were used to determine the expression levels of certain proteolytic enzymes. Complementary DNA (cDNA) microarray technique was used to determine the gene expression profiles of each of the three OSCC cell lines. The immunocytochemistry and flow cytometry results showed that the BHY cell line expresses MMP- 9 and extracellular matrix metalloproteinase inducer (EMMPRIN). The BHY cell line expressed 139 upregulated genes and 117 downregulated genes. The HN cell line expressed 214 upregulated and 262 downregulated genes. Finally, the HSC-3 cell line expressed 128 upregulated and 117 downregulated genes. There were 13 genes that were upregulated and 83 genes that were downregulated in all three OSCC cell lines. Eight pathways were upregulated and 3 pathways were downregulated in the BHY cell line. In the HN cell line 10 pathways were upregulated and 9 were downregulated. Finally, in the HSC-3 cell line 3 pathways were upregulated and 6 pathways were downregulated. MMP-1, MMP-9 and EMMPRIN appear to play a role in the invasion of OSCC to bone. KIAA, was upregulated in our study in the HN cell line. Upregulation of KIAA may be involved in the late stage of OSSC. PAM gene was the only downregulated gene in the HN cell line. The TM4SF10 gene was downregulated in all three OSCC cell lines. The COL1A2 and COL4A1 genes were upregulated in the HSC-3 cell line and they may cause the lymph node metastasis of OSCC. Based on this study OSCC involves multiple molecular genetic events that take place in many chromosomes and genes. Upregulation of the Jak-STAT signaling pathway may be involved in bone invasion of OSCC. The MAPK signaling pathway was upregulated in all three OSCC cell lines.

Feline herpesvirus 1 (FHV-1) is a linear double-stranded DNA virus that causes approximately 50% of the upper respiratory tract infections and produces the most severe respiratory disease in domestic cats. Primary ocular infection, as occurs in humans with the related virus, herpes simplex virus type 1 (HSV-1), consistently produces conjunctivitis and minimal corneal involvement; however, clinical manifestations of disease due to repeated recrudescence involve the cornea and can potentially lead to blindness. Vaccines only produce partial protection from clinical disease, and antiviral medications approved for treatment of HSV-1 in humans are only minimally effective for treatment of the chronic cases in cats. Therefore, RNA interference (RNAi), a RNA-guided gene regulatory mechanism that is found in a variety of eukaryotic organisms and provides anti-viral immunity in plants, was used as a therapeutic method for prevention of FHV-1 infection in feline cells. Previous RNAi experiments in related herpesviruses demonstrated that the DNA polymerase gene and a gene coding for a viral attachment protein are effective targets for inhibiting herpesvirus replication. Therefore, a region coding for highly conserved amino acid motifs of herpesvirus DNA polymerase genes, which is unique to each viral species and lacks DNA sequence drift for alphaherpesviruses, including herpes simplex virus type 2 and also feline herpesvirus as shown in this study, was targeted by RNAi. The attachment proteins for FHV-1 are unknown, but the highly conserved glycoprotein D (gD) gene was also chosen as a target for this study. HSV-1 gD is an essential receptor-binding polypeptide and is necessary for penetration of the virus into cells. FHV-1 gD, an envelope protein, is an inducer of virus-neutralizing antibodies and may play an important role in the restriction of the host range of the virus to feline cells. Two synthetic siRNAs targeting the DNA polymerase gene and the gD gene of FHV-1 were effective in knockdown of their intended targets by 69-83% for DNA polymerase mRNA and 77-87% for gD mRNA, determined by quantitative real-time RT-PCR. Based on flow cytometry results, glycoprotein D mRNA knockdown decreased gD cell surface expression by 27-43%, and DNA polymerase mRNA interference decreased cell surface FHV-1 glycoprotein expression by 29-71%. Knockdown of the mRNAs from these genes also resulted in decreased infective virus in vitro, with decreased viral replication by 83-96% by DNA polymerase RNAi and 77-84% by gD RNAi, determined by plaque assays. Interference of each mRNA also resulted in a decrease in the amount of the opposite mRNA, independent of interferon β production. These results indicate that FHV-1 glycoprotein D, like its homolog in HSV-1, is essential for in vitro replication and is likely involved with viral attachment and/or penetration, and both this gene and the FHV-1 DNA polymerase gene are suitable targets for RNAi anti-viral treatment. This study lays the groundwork for potential in vivo investigations in cats. Such studies may provide unique insights into the prevention/treatment of herpesvirus infections by RNAi.

The purpose of this study was to determine how normal hearing adults (NHA), normal hearing children (NHC) and children wearing cochlear implants (CI) differ in the perceptual weight given cues for fricative consonant and voiceless stop consonant continua. Ten normal-hearing adults (NHA), eleven 5-8-year-old normalhearing children (NHC) and eight 5-8-year-old children wearing cochlear implants (CI) were participants. For fricative consonant perception, the /su/-/∫u/ continua were constructed by varying a fricative spectrum cue in three steps and by varying a F2 onset transition cue in three steps. For voiceless stop consonant perception, the /pu/- /tu/ continua were constructed by varying a burst cue in three steps and a F2 onset transition cue in three steps. A quantitative method of analysis (ANOVA model) was used to determine cue weighting and measure cue interaction. For the fricative consonant, both NHC and NHA gave more perceptual weight to the frication spectral cue than to the formant transition. NHC gave significantly less weight to the fricative spectrum cue than NHA. The weight given the transition cue was similar for NHC and NHA, and the degree of cue interaction was similar between two groups. The CI group gave more perceptual weight to the fricative spectrum cue than to the transition. The degree of cue interaction was not significant for CI. For the voiceless stop consonant, both NHC and NHA gave more perceptual weight to the transition cue than to the burst cue. NHC gave proportionately less weight to the transition cue than NHA. The weight given the burst cue and the degree of cue interaction were similar between NHC and NHA. The CI group gave more perceptual weight to the transition cue than to the burst cue, and there was no significant difference between children wearing cochlear implants and normal hearing children group; however, the degree of cue interaction was not significant for CI. These results indicated that all groups favored the longer-duration cue to make phonemic judgments. Also there were developmental patterns. The CI group has similar cue weighting strategies to agematched NHC, but the integration of the cues was not significant for either fricative or voiceless stop consonant perception.

Herpetic stromal keratitis (HSK) is an immunopathological and tissue destructive corneal lesion caused by herpes simplex virus (HSV) infection, which induces an intense inflammatory response and finally leads to blindness. Accumulating evidence using the murine model has shown that Th-1 phenotype CD4+ T cells orchestrating the inflammation mainly contribute to the immunopathological reaction in HSV-1 infected cornea. However, prior to CD4+ T cell infiltration into corneal lesions, various innate immune cells recruit and produce numerous inflammatory and angiogenic molecules into the corneal stroma those in turn drive the corneal immunopathology. The first part (Part I) of this dissertation focuses on the understanding of HSV-1 induced immunoinflammatory processes in the cornea and trigeminal ganglia including the secondary lymphoid tissues. The next three parts (Part II-IV) focus on different inflammatory and anti-inflammatory mechanisms that are activated following virus infection in the cornea. Results in Part II evaluate the role of various Toll-like receptors (TLRs) in driving the early inflammatory events that occur in the HSV infected corneas. Thus, this part demonstrates that mainly TLR2 and to a lesser extent TLR9 ligand activity of HSV is important for driving SK pathology. Results of the third section show that HSV infection results in the up regulation of IL-23 that is needed for the survival of pathogenic Th-17 cells. The data show that mice lacking IL-23 were more susceptible to SK lesions and that the heightened lesion severity was attributed to higher IL-12 production and Th1 immune response in such animals. The fourth section describes the relative role of IL-10 and natural regulatory T cells (nTregs) as two independent anti-inflammatory mechanisms that are needed for controlling HSK lesions. In this study, experiments were designed to understand the mechanisms involved in the induction and regulation of immunopathology in HSK and modulation of such processes could be useful in designing therapeutic for HSK.

Objective Because the public health nutrition workforce may be in a state of transition, this study had three purposes: 1) describe the US public health nutrition workforce; 2) examine a new position class, breastfeeding peer counselor; and 3) determine if retirement intention of public health nutrition personnel can be predicted based on personal and workplace factors. Methods Secondary data analysis of the national research dataset of the 2006-07 Public Health Nutrition Workforce Survey was conducted (n=10,683, response rate 80.0% for overall survey; research dataset n=9,923). Subjects were personnel in nutrition professional/paraprofessional positions working in nutrition programs under the purview of the official health agency and who agreed to release their data for research purposes. Results Over one-quarter (28.0%) of respondents were in positions with a population/systems focus, while 67.5% were in client-focused, direct care positions. Two-thirds (67.0%) practiced primarily in the core public health function of assurance. Approximately 10% (11.3%) of personnel were breastfeeding peer counselors. The majority (52.6%) of breastfeeding peer counselor positions were part-time and 20.3% were contracted. Nearly half (42.0%) did not receive employee benefits. Close to one-quarter (23.9%) of the overall workforce intended to retire within 10 years. There were significant differences in both personal and workplace factors for intention to retire for personnel 45 years and older. Age category, years of experience in nutrition/dietetics and public health nutrition, agency of employment, vacation and retirement employee benefits, percent of work time spent in direct client services, full-time/part-time status, and US DHHS Region correctly predicted retirement intention 75.0% of the time. Conclusions The majority of respondents worked in client-focused positions which could indicate a potentially inadequate proportion of personnel available for assuring population health. Breastfeeding peer counselors constitute a noteworthy proportion of the overall workforce. That many positions are part-time or contracted and do not receive employee benefits could indicate inadequate funding for this position class. ‘Graying’ of the public health nutrition workforce appears to be an important concern. Results can be used to evaluate organizational characteristics for workforce succession planning and forecasting.