HLA-DR1 transgenic (DR1 Tg) mice provide a model for evaluating the breadth and specificity of CD4 T cell responses that may develop in humans following influenza infection or vaccination. Recent studies identified a tremendously broad HLA-DR1-restricted CD4 T cell responses in DR1 Tg mice infected intranasally with influenza A/New Caledonia/20/99 (NC). In this study, our goals were to characterize B cell responses after NC infection in DR1 Tg mice and establish the correlation between B cell responses and CD4 T cell responses in this system. Influenza-specific B cell responses following virus administration were analyzed in DR1 Tg mice and in the genetically matched H-2b strain C57BL/10J (B10). Following intranasal (i.n.) NC infection, B cell responses in B10 mice featured strong IgG2b and IgG2c production and were typical of previously described B cell responses to a variety of mouse-adapted influenza strains. In contrast, B cell responses in DR1 Tg mice followed delayed kinetics and were strongly skewed to IgG1 production, suggesting the Th2 polarization of CD4 T cell responses. The different antibody isotype profile in DR1 Tg mice compared to B10 mice was evident in antibody secreting cells (ASCs) frequencies and in circulating Abs levels. Surprisingly, although DR1 Tg mice had lower influenza-specific Abs levels, they exhibited higher neutralizing Abs titers early in the response. B cell responses following intranasal infection of influenza A/Puerto Rico/8/1934 (PR8) or intramuscular vaccination of inactivated NC in DR1 Tg mice were different from the observed IgG1 bias after i.n. NC infection. After i.n. PR8 infection, B cell responses were similar in DR1 Tg mice and B10 mice, characterized by predominant IgM/IgG3 production. Additionally, following intramuscular administration of inactivated NC, B cell responses were skewed towards IgG2c production in both DR1 Tg mice and B10 mice, suggesting the Th1 polarization of CD4 T cell responses. A mechanistic understanding of IgG1/Th2 biased B cell responses and better neutralizing Abs production in DR1 Tg mice following i.n. NC infection may have implications for the optimal control of influenza infection.

Respiratory syncytial virus (RSV) infection causes ~64 million cases of respiratory disease and 200,000 deaths annually worldwide, yet there is no broadly effective prophylactic or treatment regimen. RSV can produce acute respiratory illness in patients of all ages but strikes the age extremes, infants and the elderly, with highest frequency presumably due to innate immune deficiencies. A higher morbidity and mortality has been reported for the elderly above 65 years of age, which has been attributed to immune senescence. Efforts to generate an effective vaccine have thus far been unsuccessful. The innate immune system provides the first line of defense against viral pathogens with a repertoire of anatomical barriers, phagocytic immune cells, pattern recognition receptors (PRRs) and antiviral cytokines like interferons (IFNs). The precise mechanism of subversion of innate immunity in young and aged is poorly understood. A better understanding of innate immune pathways is expected to aid in the development of appropriate vaccines or prophylactics for these high-risk groups. Previously, the RSV nonstructural protein 1 (NS1) was shown to antagonize IFN responses by disrupting components of the innate immune system, although the mechanism is not well defined. We hypothesized that NS1 targets constituents of the PRR pathways to evade innate immunity and thus ensure viral survival. Using microscopy and co-immunoprecipitation assays, we found that NS1 localizes to the mitochondria and binds to the mitochondrially associated adaptor protein MAVS, thus preventing MAVS interaction with the RNA helicase, RIG-I. Expression of NS1 was also correlated with upstream IFN-response regulator, LGP2, and its expression was inducible in the absence of a viral infection. Tetracycline-inducible expression of recombinant NS1 in a cell model also promoted viral replication and emphasizes the key contribution of NS1 to RSV survival. Through this study, we demonstrated a mechanism for RSV NS1 in the disruption of early innate responses through mitochondrial localization and alteration of the RLH signaling. Whereas the above studies showed the importance RSV-induced innate immune pathways, whether the expression and signaling of innate immune pathways were adversely affected upon RSV infection in the high-risk groups remains unknown. Since elderly individuals are at an increased risk for severe bronchiolitis and RSV-induced pneumonia, often resulting in hospitalization and medical intervention and adaptive immune cell functionality and responsiveness reportedly decline with age, we hypothesized a similar age-related deterioration of the innate antiviral system. In this investigation, we used an aged mouse model to correlate age-associated changes in innate immune gene expression with RSV pathology. Of 84 antiviral genes examined, five genes including RIG-I, IFNAR1, TLR8, IL-1Β, and osteopontin (OPN) were associated with both age and infection. In response to RSV infection, aged mice had delayed induction of antiviral genes and diminished ability to secrete IL-6 in response to TLR7/8 agonist in primary alveolar macrophages. Lungs from aged, RSV-infected mice had increased cellular infiltration and prolonged infection as compared to young mice. In summary, age-related decline in expression and functionality of antiviral defenses were correlated with enhance RSV-induced lung disease in aged mice. In the absence of infection, aged mice chronically overproduced IL-1Β and OPN relative to young mice. Upon infection, aged mice had impaired ability to secrete higher levels of IL-1Β and mucus. In contrast, OPN secretion remained high and prolonged in aged mice throughout infection. The age-related decline in host antiviral gene induction and delayed cytokine production correlated with enhanced disease pathology. Using a transgenic strain of mice deficient in OPN (OPN-KO), we observed greater resistance to RSV and enhanced secretion of mucus, but unaltered cellular infiltration into the lungs. Therefore, OPN overproduction and defective mucus production likely contribute to pathology in aged mice. These findings demonstrate that RSV targets the innate virus recognition and antiviral cytokine activation pathways but also that the antiviral defense system is significantly affected by age. Consequently, efforts to generate vaccines or develop therapies that stimulate IFN induction may prove unsuccessful in the elderly given that RSV virulence factors and age weaken these responses. This study contributes to our understanding of how aging relates to the RSV subversion of the host antiviral response and should help with the development of better antiviral therapies suited to the growing elderly population.

Across three studies, the role that testosterone plays in how individuals respond psychologically, behaviorally, and physiologically to status challenges was investigated. Preliminary Studies 1 focused on how testosterone related to physiological and psychological responses to a medical threat. Preliminary 2 replicated the psychological effects observed in Preliminary Study 1. Study 3 examined how experimentally manipulated testosterone levels corresponded to responses to a socially judged physical endurance task across all three response types. Preliminary Study 1 examined the relationship between testosterone and conscious evaluations of and physiological reactions to a health threat. Participants were diagnosed with a fictitious enzyme deficiency before rating their views of the deficiency, as well as providing saliva samples before and after diagnosis. Basal testosterone was negatively associated with the belief that one actually had the deficiency, despite the diagnosis. Testosterone was also positively associated with a greater increase in salivary cortisol levels following the diagnosis. Self-reported anxiety was found to be positively associated with evaluating the deficiency as threatening. Preliminary Study 2 replicated the findings observed in Preliminary Study 1 regarding conscious evaluations of a medical threat. Using the same experimental manipulation, testosterone was again found to be negatively associated with ratings of the enzyme deficiency. In Preliminary Study 2, high levels of testosterone were associated with viewing the deficiency as less serious and viewing medical conditions, in general, as less threatening. Study 3 used a transdermal administration procedure to artificially elevate individuals’ testosterone levels before completing a socially evaluated task. Participants who received the testosterone administration showed greater physiological responses to the task, including cardiovascular responses and cortisol responses, compared to the placebo group. Unlike Preliminary Studies 1 and 2, Study 3 did not show any effect of testosterone on conscious evaluations of the task nor behavioral measures of performance. Taken together, the three studies highlight the different ways in which testosterone is related to responding to social threats. Testosterone appears to be associated with mobilizing physiological systems to theoretically facilitate behavioral responses to status threats. Testosterone also appears to be negatively associated with consciously evaluating certain types of threats. / text

An effective vaccine to protect against HIV-1/AIDS remains elusive due to the extensive mechanisms employed by the HIV-1 virus to evade immune attack. Highly potent broadly neutralizing antibodies isolated from chronically infected individuals, however, show that such relevant antibodies can be naturally produced, implying that their elicitation through vaccination is a realistic possibility. These broadly neutralizing antibodies target different regions on the trimeric spikes formed by three protomers of the envelope (Env) protein. Each Env protein is comprised of the gp120 surface subunit in non-covalent association with the gp41 transmembrane subunit. Four regions have been identified: the CD4 binding site, the V1/V2 segment and the V3/glycan area all on the gp120 subunit as well as the MPER segment on the gp41 subunit. This dissertation focuses on the gp41 MPER segment given its highly conserved amino acid sequence among all HIV-1 clades and viral strain isolates and essential function in Env-mediated fusion and HIV entry. Of note, the MPER segment contains several adjacent epitopes targeted by broadly neutralizing antibodies, suggesting that the immune system is capable of producing neutralizing antibodies against this specific region. Analysis of both clade B and C MPER segments shows them to be L-shaped, consisting of two  helices separated by a hinge. We have found that the hinge region of the MPER segment provides the conformational flexibility necessary for the Env-mediated hemifusion and fusion processes. A significant reduction in virus infectivity is observed when the hinge region is disrupted by introduction of two amino acid mutations that eliminate -helical capping residues and the tandem hinge joints. The importance of the hinge region of the MPER segment is further supported by the action of four MPER-specific neutralizing antibodies 2F5, 4E10, 10E8 and Z13E1. These neutralizing antibodies block virus infection by disrupting MPER hinge-related function.

ISG15 is an interferon-induced and anti-viral ubiquitin-like protein (Ubl). Ube1L, UbcH8, and Herc5 have been identified as the E1-E2-E3 enzymes for ISG15 conjugation, and, like ISG15, their expression is induced by type I interferons. Although Herc5 is the major E3 for ISG15, over 300 proteins have been identified as ISG15 target proteins in interferon-stimulated cells. In this work, I address two aspects of the human ISG15 conjugation system: 1) the specificity of the Ube1L-UbcH8 interaction and 2), the basis of substrate recognition by Herc5. Regarding the selection of UbcH8 by Ube1L, my experiments show that although UbcH8 had been reported to function as an E2 for both Ub and ISG15, UbcH8 is preferentially activated by Ube1L compared to Ube1 (E1[superscript Ub]). The basis of this preference is a result of specific interactions between the ubiquitin-fold domain (UFD) of Ube1L and the amino-terminal [alpha]1 helix and [beta]1 [beta]2 region within UbcH8. Examination of the interferon-induced and transfected expression levels of UbcH8, combined with the kinetic constants, suggest that UbcH8 is unlikely to function as a Ub E2 in most cell lines. In examining the selection of target proteins by Herc5, I show that the range of substrates extends far beyond the proteins identified in proteomics studies and includes many exogenously expressed foreign proteins. Furthermore, I show that ISG15 conjugation is restricted to newly synthesized pools of proteins and Herc5 is associated with polyribosomes. I propose a model for ISGylation in which Herc5 broadly modifies newly synthesized proteins in a co-translational manner and suggest that, in the context of an interferon-stimulated cell, newly translated viral proteins may be primary targets of ISG15. Consistent with this, I show that ISGylation of human papillomavirus (HPV) L1 capsid protein has a dominant-inhibitory effect on the infectivity of HPV16 pseudoviruses. These discoveries have greatly increased our understanding of the mechanism of ISG15 pathway and provide a framework for establishing an in vitro ISG15 conjugation system and further examination of the anti-viral function of ISG15. / text

This paper seeks to chronicle the different ways marketers have successfully defended and/or defeated legal challenges that seek to stump their creativity. Previous research, existing laws and regulating agencies are surveyed to define the current lay of the land. Ten unique cases are analyzed to discover over ten alternate responses that brand ambassadors have either successfully or unsuccessfully provided in the face of legal crisis. The findings are summarized, and further research avenues are suggested with the aspiration that up-and-coming brands gain insights from lessons learned in the past. / text

Human activities that disrupt soil properties are fundamentally changing ecosystems. Soil degradation decreases microbial abundance and activity, leading to changes in nutrient availability, soil organic matter, and plant growth and establishment. Land use and land cover change are widespread and increasing in semiarid regions of the southwestern US, which results in reductions of native plant and microbial abundance and community diversity. Here we studied the effects of soil degradation and amendments (biochar and woodchips) on microbial activity, soil carbon and nitrogen availability, and plant growth of ten semi-arid plants species native to the southwestern US. Results show that woodchip amendments result in poor overall plant growth, while biochar amended soils promoted plant growth when soil quality was reduced. Additionally, amendments had a strong influence on microbial activity, while the presence and species identity of plants did not. Biochar amended soils led to increases in the potential activities of enzymes involved in the degradation of carbon, nitrogen, and phosphorus rich substrates. Woodchips, caused an increase of potential activity in enzymes involved in the degradation of sugar and proteins. These results show that microbes and plants respond differently to soil treatments and suggest that microbial responses may function as earlier indicators of the success of re-vegetation attempts.

This study explores the cultural dimension of translating children's and adolescent literature. Framed within the theories of cultural studies, translation studies, Baktinian dialogism, and reader response theories, this study is three-fold: (1) a content analysis is conducted to identify the cultural and linguistic shifts in the translated books and the strategies utilized by the translators for making those shifts, (2) the responses of the source-text (ST) and the target-text (TT) readers are compared; (3) the potential relationship between the translation strategies and the reader responses are inferred based on the findings from (1) and (2). The expected findings are: (1) adept use of various translation strategies helps the TT readers recognize themes as similar as the ST readers; (2) some interventions may create deviating responses in the TT readers as compared with the ST readers; (3) some unique responses by either the ST or the TT readers may be as a direct result of cultural differences more than the translation strategies. The implication section provides recommendations to publishers, translators, educators, parents, teacher educators, and researchers, and suggestions for further research.

The purpose of this study was to explore and understand the emotional responses of selected high school readers when they engage in retrospective miscue analysis.Several data sets were collected through audio and video taping of interviews, readings, and individual and group sessions. Analysis of the data involved the use of In Depth procedure of miscue analysis to examine readers' meaning construction, grammatical patterns, and word substitution similarities. Results from the miscue analysis sessions were used to engage the students in retrospective miscue analysis (RMA). RMA consisted of engaging readers to reflect and evaluate the reading process and strategies by analyzing their miscues. Their emotional responses during the RMA sessions were examined and analyzed to describe patterns in readers' revalued voices. Martin and White's (2005) appraisal theory was used to analyze student's emotional responses. Appraisal theory is based on Halliday's systemic functional linguistic view of language.Research findings indicated that readers became adept at articulating their own strategies, fine tuned their own affective stance about reading and used what they know about miscues and reading to better themselves as readers. Their emotional responses towards reading also changed over time as students began to use linguistic resources to agree, disagree, critique, and position their listeners to their own assessments and adapted their own revalued voice about who they are as readers. Readers' miscues also showed that they began to focus more on making meaning, thus improving their reading.

Persons with aphasia (PWA) have language difficulties and their Quality of Life (QoL) has most likely been affected. Because of their loss of language abilities, it is difficult to use PWA in QoL studies. This can lead to the use of proxy responses (a person who answers for the PWA). The aim of this study was to compare the rankings from QoL questionnaires for PWA with the rankings from their spouses/caregivers (i.e., proxyresponse). We also wanted to find out which of the 11 aspects of life PWA andspouses/caregivers ranked as the most respectively the least impacted ones. A totalnumber of 57 persons participated in the study. The participants consisted of two groups, one group with PWA and one group of their spouses/caregivers. A questionnaire-based cross-sectional survey completed via a face-to-face interview was used to collect data from both groups. The results showed that there was poor internal consistency and a weak correlation between the two groups. The two groups ranked different aspects of life as “most impacted” and “least impacted”. The PWA ranked Vocation/Occupation as the “most impacted” and Family life as the “least impacted”aspects of life. The spouses/caregivers ranked Overall ability to communicate as the“most impacted” and Ability to self-care as the “least impacted” aspects of life. The results indicate that proxy responses may not be appropriate and should be interpreted with caution in QoL studies with PWA.