Herpes simplex viruses (HSV) establish latent infections in sensory neurons of their host and are maintained in this state by little understood mechanisms that, at least in part, are regulated by signalling through nerve growth factor (NGF) and its receptor tropomyosin related kinase, trkA. Previous studies have demonstrated that Zhangfei is a transcriptional factor that is expressed in differentiated neurons and is thought to influence HSV replication and latency. Zhangfei, like the HSV trans-activator VP16 and Luman, binds the ubiquitous nuclear protein host cell factor (HCF) inhibiting the ability of VP16 and Luman to initiate HSV replication. <p>Recently, Brn3a, another neuronal factor thought to influence HSV latency and reactivation was found to possess an HCF-binding domain and could potentially require HCF for activity. The neuronal POU IV domain protein, Brn3a, among its many regulatory functions has been described as an enhancer of the NGF receptor trkA, during development in mouse. I therefore investigated the possible link between Brn3a, TrkA, NGF signaling, HCF, Zhangfei and HSV-1 latency and reactivation. I hypothesized that Zhangfei would also suppress the ability of Brn3a to activate the expression of TrkA and that this would have an impact on NGF-TrkA signaling and, consequently on HSV-1 reactivation from latency.<p>My first study determined which Brn3a/trkA promoter interactions were important for trkA transcription. I constructed a plasmid that contains 1043 base pairs of genomic sequences that extend from 30 nucleotides upstream of trkA coding region. In contrast to previous data, a short 190 bp region that lies proximal to the trkA initiation codon was sufficient for Brn3a trans-activation in NGF-differentiated PC12, Vero and human medulloblastoma cells. At least two portions of the 190 bp fragment bind to Brn3a. In addition, Brn3a increased endogenous levels of trkA transcripts in PC12 cells and initiated trkA expression in medulloblastoma cells, which normally do not express trkA. <p>The second step was to determine the effects of HCF and Zhangfei association with Brn3a on trkA trans-activation. I found that Brn3a required HCF for activating the trkA promoter and that Zhangfei has a suppressive effect over Brn3a-trkA activation in non-neuronal cells. In sympathetic neuron-like NGF-treated PC12 cells, Zhangfei did not suppress the ability of Brn3a to activate the TrkA promoter, however, Zhangfei was able capable of inducing the expression of TrkA in the absence of Brn3a. Both Brn3a and Zhangfei induced the expression of endogenous trkA in PC12 cells.<p>Since Vero and PC12 cells are not from human origin I wanted to examine the ability of Zhangfei to induce trkA transcription in medulloblastoma cells, that because of its tumor nature do not express trkA. TrkA transfections in these cells have shown to drive them to cell arrest or apoptosis. Since Zhangfei is not express in medulloblastoma tumors I then used ONS-76 medulloblastoma cells as a model to determine Zhangfeis envolvement in the NGF-trkA signaling pathway.<p> I show herein that in ONS-76 medulloblastoma cells resveratrol, an inducer of apoptosis and differentiation, increased the expression of Zhangfei and trkA as well as Early Growth Response Gene 1 (Egr1), a gene normally activated by NGF-trkA signalling. ONS-76 cells stop growing soon after treatment with resveratrol and a portion of the cell undergo apoptosis. While the induction of Zhangfei in resveratrol-treated cells was modest albeit consistent, the infection of actively growing medulloblastoma cells with an adenovirus vector expressing Zhangfei mimicked the effects of resveratrol. Zhangfei activated the expression of trkA and Egr1 and caused these cells to display markers of apoptosis. The phosphorylation of Erk1, an intermediate kinase in the NGF-trkA signaling critical for differentiation, was observed in Zhangfei infected cells, supporting the hypothesis that Zhangfei is a mediator of trkA-NGF signaling in theses cells leading either to differentiation or apoptosis. Binding of HCF by Zhangfei did not appear to be required for this effect as a mutant of Zhangfei incapable of binding HCF was also able to induce the expression of trkA and Egr1. <p>In in vivo and in vitro models of HSV-1 latency, the virus reactivates when NGF supply to the neuron is interrupted. Based on the above evidence Zhangfei, in HSV-1 latently infected neurons, would have the ability to prolong a state of latency by inducing trkA expression allowing the activation of NGF-trkA signaling pathway. Since NGF is produced by many cell types it is possible that reactivation is triggered not by a decrease in NGF but by a down-regulation of TrkA expression.Therefore, if Zhangfei expression is suppress the trkA signaling could be interrupted or shifted towards apoptosis signaling, this would allow neuronal HCF-binding proteins like Luman, which can activate HSV IE expression, to initiate HSV IE expression and subsequently viral replication.

This study measured knowledge and attitudes about genital herpes and human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) in a sample of future teachers from the College of Education at Oregon State University. The objectives of the study were 1) to determine if students possess accurate knowledge about the two diseases; 2) to measure attitudes toward people with the two diseases; 3) to assess the relationship, if any, between knowledge and attitudes; 4) to compare knowledge and attitudes about genital herpes with knowledge and attitudes about HIV/AIDS; and 5) to compare knowledge and attitudes about genital herpes in 1990 to data from a similar study conducted in 1984. A convenience sample of 150 students was obtained from undergraduate classes in the College of Education during Spring Term 1990. Subjects completed self-administered questionnaires about either genital herpes or HIV/AIDS during class time. Data were gathered using four instruments: A knowledge test, two attitude measures, and a demographic data questionnaire. Statistical tests used for data analysis were chi square, Pearson's correlation coefficient, Student's t-test, two-way analysis of variance (ANOVA), and repeated measures ANOVA. The significance level was .05. Knowledge scores on the HIV/AIDS test were quite high (mean score 88% correct), while the mean genital herpes knowledge score was relatively low (62% correct). Attitudes toward people with both genital herpes and HIV/AIDS were relatively accepting, but subjects were significantly more accepting toward people with genital herpes. The least accepting responses toward people with either disease occurred in regard to potentially sexual situations (e.g. dating, marriage). There was no gender difference in attitudes toward people with either disease. Attitudes were more positive in response to a vignette of a college student followed by a questionnaire, compared to responses made to a questionnaire only. Correlations were found between more knowledge and more accepting attitudes about both diseases. Finally, genital herpes knowledge scores were higher (mean score 62% correct) than scores from a similar study of genital herpes conducted in 1984 (mean score 57% correct). Attitudes toward people with genital herpes were more accepting in the 1990 sample than were attitudes in the 1984 sample. All findings reported here are statistically significant. Recommendations for future research and education among future teachers concerning sexually transmitted diseases (STD's) include 1) development of methods to transmit accurate information about STD's by personalizing these diseases and relating them to college students' experiences; 2) a research focus upon attitudes and perceptions about STD's among future teachers, including the issue of homophobia, and how these relate to behavior; and 3) thorough teacher preparation about STD's, focusing on accurate knowledge and impartial attitudes that allow this topic to be addressed effectively in the classroom. Future research among the general college student population should address 1) the relationship between knowledge, attitudes, perceptions, and behavior concerning STD's; 2) potential differences in responses made to a vignette followed by a questionnaire, compared to a questionnaire only; 3) students' source(s) of information about STD's, and level of trust in "scientific authority"; 4) possible interactions between religious influence and attitudes about STD's; 5) the existence of a stereotype of HIV/AIDS as a gay, male disease, and how this might affect attitudes and perceptions; 6) differences between males and females in terms of attitudes, especially with regard to homophobia; 7) the effectiveness of personalizing STD education to increase knowledge about and perceived susceptibility to STD's; 8) the interaction between societal values and personal values, and their effect on attitudes about STD's and sexual behavior. / Graduation date: 1991

L'estudi de la utilització del sistema immunològic de l'hoste pels virus com a patògens per al seu propi benefici fou el principal objectiu d'aquest treball. Concretament, en aquest treball de tesis es van analitzar dues espècies d'herpesvirus virulents en humans, l'Herpes Simplex Virus-1 (HSV-1) i l'Epstein Barr Virus (EBV). L'entrada d'ambdós virus a les cèl.lules és mediada per receptors del sistema immunològic. Es va estudiar la interacció dels respectius receptors cel.lulars, Herpes Virus Entry mediator A (HveA), i el receptor de complement 2 (CR2), amb les glicoproteïnes virals que s'hi uneixen, i amb llurs lligands naturals, els quals participen en la defensa de l'hoste. Es va caracteritzar la interacció dels receptors HveA i CR2 amb els seus lligands fent servir proteïnes recombinants o purificades de sèrum; en el cas de HveA ens vam centrar en la localització del lloc d'unió de cada lligand al receptor. En el cas de CR2, es va analitzar la cinètica d'unió dels seus lligands naturals. Per a ambdós receptors, es va analitzar si la unió de la proteïna viral al receptor podria interferir i/o modular-ne la unió dels seus lligands naturals. Els resultats es van analitzar dins del marc de la resposta immunològica de l'hoste mediada pel receptor cel·lular, i en relació al possible paper modificador d'aquesta resposta per part de la proteïna viral. Per a facilitar el nostre estudi sobre HveA, es van cercar nous lligands peptídics d'aquest receptor, utilitzant llibreries aleatòries de pèptids expressades en el fag M13. Es van aïllar dos pèptids, i es va estudiar la seva interacció amb el receptor i la seva capacitat d'inhibir l'entrada del virus a les cèl·lules, és a dir, com a possibles agents terapèutics. / Our goal in the present work was to study the manipulation of the host immune system by an infecting virus to its own benefit. Specifically, we studied two herpes viruses; Herpes Simplex Virus-1 (HSV-1) and Epstein Barr Virus (EBV), which infect humans. Entry of both viruses into the cell is mediated by the interaction of a specific viral surface glycoprotein with two receptors that participate in the host immune response, the Herpes Virus Entry mediator A (HveA), and complement receptor 2 (CR2). We studied the interaction of these receptors with the viral glycoproteins as well as their host ligands. The latter play a role in the immune response of the host. We characterized these interactions by using recombinant as well as serum-purified proteins. Our study of HveA sought to localize the specific binding site of each ligand on the receptor, while that of CR2 consisted in the kinetic analysis of its interaction with its ligands. We also analyzed whether binding of the viral glycoprotein to each receptor would interfere or modulate its interaction with its host ligands. Our results were analyzed in the context of the role of these receptors in the host immune response, and specifically whether the viral proteins studied undermined the host's ability to defend itself from infection. To study the relationship between HveA, its natural ligands, and the viral proteins involved in HSV entry, we also screened two phage-displayed combinatorial peptide libraries for peptide ligands of a recombinant form of HveA. We isolated two peptides, and studied their interaction with HveA as well as their ability to block HSV entry into HveA-bearing cells.

Fourier transform infrared (FTIR) microscopy has shown promise as an analytical tool for detecting changes in cells and tissues, such as those due to viral infection, apoptosis induction or malignancy. In many cases, diagnosis via FTIR microscopy can be undertaken on a timescale shorter than that required for other physical or histological techniques. In this work we have used FTIR microscopy to study Vero cells that have been infected with herpes simplex virus (type I) and adenovirus. We have studied cellular samples at various time intervals following exposure to the virus. Several spectral regions were identified that allow discrimination between infected and uninfected Vero cell samples at 24 hours post exposure to both HSV1 and adenovirus. Spectral features were also identified that could be used to discriminate infected cells within 2-6 hours after exposure to both viruses. FTIR microscopy is therefore a useful tool for following the kinetics of viral infection in the 2-24 hours time range, at least at the levels of infection used in this study. In a second type of study, FTIR microscopy was used to study apoptosis induction in acute lymphoblastic leukemia T-cells. Apoptosis was induced in T-cells in three different ways. We show that FTIR microscopy can be used to distinguish T-cells in the early stages of apoptosis from normal cells. We also provide data that may suggest that FTIR microscopy can distinguish cells that have undergone apoptosis via different pathways. For most of the FTIR microscopic studies on cellular samples we have focused on the collection of spectral data in the 1500-800 cm-1 region. Spectra were collected for control cells and variously treated cells. The two sets of cells were then analyzed statistically using: 1) pair-wise comparison, 2) logistic regression, 3) partial least square regression, 4) principle component fed linear discriminant analysis and 5) hierarchical cluster analysis. The statistical analyses rigorously quantify to what extent treated and untreated cells can be distinguished. Since different statistical methods give differing results for the same data, it is important the right statistical method should be applied. The basis for these differences is discussed.

Herpes simplex viruses (HSV) establish latent infections in sensory neurons of their host and are maintained in this state by little understood mechanisms that, at least in part, are regulated by signalling through nerve growth factor (NGF) and its receptor tropomyosin related kinase, trkA. Previous studies have demonstrated that Zhangfei is a transcriptional factor that is expressed in differentiated neurons and is thought to influence HSV replication and latency. Zhangfei, like the HSV trans-activator VP16 and Luman, binds the ubiquitous nuclear protein host cell factor (HCF) inhibiting the ability of VP16 and Luman to initiate HSV replication. <p>Recently, Brn3a, another neuronal factor thought to influence HSV latency and reactivation was found to possess an HCF-binding domain and could potentially require HCF for activity. The neuronal POU IV domain protein, Brn3a, among its many regulatory functions has been described as an enhancer of the NGF receptor trkA, during development in mouse. I therefore investigated the possible link between Brn3a, TrkA, NGF signaling, HCF, Zhangfei and HSV-1 latency and reactivation. I hypothesized that Zhangfei would also suppress the ability of Brn3a to activate the expression of TrkA and that this would have an impact on NGF-TrkA signaling and, consequently on HSV-1 reactivation from latency.<p>My first study determined which Brn3a/trkA promoter interactions were important for trkA transcription. I constructed a plasmid that contains 1043 base pairs of genomic sequences that extend from 30 nucleotides upstream of trkA coding region. In contrast to previous data, a short 190 bp region that lies proximal to the trkA initiation codon was sufficient for Brn3a trans-activation in NGF-differentiated PC12, Vero and human medulloblastoma cells. At least two portions of the 190 bp fragment bind to Brn3a. In addition, Brn3a increased endogenous levels of trkA transcripts in PC12 cells and initiated trkA expression in medulloblastoma cells, which normally do not express trkA. <p>The second step was to determine the effects of HCF and Zhangfei association with Brn3a on trkA trans-activation. I found that Brn3a required HCF for activating the trkA promoter and that Zhangfei has a suppressive effect over Brn3a-trkA activation in non-neuronal cells. In sympathetic neuron-like NGF-treated PC12 cells, Zhangfei did not suppress the ability of Brn3a to activate the TrkA promoter, however, Zhangfei was able capable of inducing the expression of TrkA in the absence of Brn3a. Both Brn3a and Zhangfei induced the expression of endogenous trkA in PC12 cells.<p>Since Vero and PC12 cells are not from human origin I wanted to examine the ability of Zhangfei to induce trkA transcription in medulloblastoma cells, that because of its tumor nature do not express trkA. TrkA transfections in these cells have shown to drive them to cell arrest or apoptosis. Since Zhangfei is not express in medulloblastoma tumors I then used ONS-76 medulloblastoma cells as a model to determine Zhangfeis envolvement in the NGF-trkA signaling pathway.<p> I show herein that in ONS-76 medulloblastoma cells resveratrol, an inducer of apoptosis and differentiation, increased the expression of Zhangfei and trkA as well as Early Growth Response Gene 1 (Egr1), a gene normally activated by NGF-trkA signalling. ONS-76 cells stop growing soon after treatment with resveratrol and a portion of the cell undergo apoptosis. While the induction of Zhangfei in resveratrol-treated cells was modest albeit consistent, the infection of actively growing medulloblastoma cells with an adenovirus vector expressing Zhangfei mimicked the effects of resveratrol. Zhangfei activated the expression of trkA and Egr1 and caused these cells to display markers of apoptosis. The phosphorylation of Erk1, an intermediate kinase in the NGF-trkA signaling critical for differentiation, was observed in Zhangfei infected cells, supporting the hypothesis that Zhangfei is a mediator of trkA-NGF signaling in theses cells leading either to differentiation or apoptosis. Binding of HCF by Zhangfei did not appear to be required for this effect as a mutant of Zhangfei incapable of binding HCF was also able to induce the expression of trkA and Egr1. <p>In in vivo and in vitro models of HSV-1 latency, the virus reactivates when NGF supply to the neuron is interrupted. Based on the above evidence Zhangfei, in HSV-1 latently infected neurons, would have the ability to prolong a state of latency by inducing trkA expression allowing the activation of NGF-trkA signaling pathway. Since NGF is produced by many cell types it is possible that reactivation is triggered not by a decrease in NGF but by a down-regulation of TrkA expression.Therefore, if Zhangfei expression is suppress the trkA signaling could be interrupted or shifted towards apoptosis signaling, this would allow neuronal HCF-binding proteins like Luman, which can activate HSV IE expression, to initiate HSV IE expression and subsequently viral replication.

No description available.

El objetivo del presente estudio fue conocer la prevalencia del Virus Herpes Bovino 1 (VHB-1), agente causal de la Rinotraqueitis Infecciosa Bovina (RIB) en bovinos criollos de crianza extensiva de los distritos de Coracora, Chumpi, Puyusca y Pullo de la provincia de Parinacochas, Ayacucho. Con esta finalidad se consideraron 469 muestras de sueros bovinos procedentes de 25 hatos para la detección de anticuerpos neutralizantes mediante la prueba de neutralización viral. El 67.59 ± 4.24% (317/469) de las muestras presentaron anticuerpos neutralizantes con títulos entre 1:2 a> a 1:256. El 100% de los hatos muestreados tuvieron animales seroreactores. La prevalencia del virus fue similar en los animales de los 4 distritos estudiados. Este estudio reporta la presencia del VHB-1 en bovinos criollos de la provincia de Parinacochas, con una prevalencia superior a lo descrito en bovinos de las principales cuencas lecheras del país. / The seroprevalence of Bovine Herpes Virus-1, the causative agent of Infectious Bovine Rinotracheitis (IBR), in criollo bovines from districts of Coracora, Chumpi, Puyusca and Pullo of the Parinacochas Province, Ayacucho was carried out. Four hundred sixty nine serum samples from twenty five herds were tested by virus neutralisation test to detect neutralizing antibodies. The 67.59 ± 4.24% (317/469) of the samples had antibodies against BHV-1. The seroprevalence of the virus was similar in the animals from the 4 district studied. The antibodies titers ranged from 1:2 to> 1:256. All the sampled herds had seroreactive animals. This study report the presence of the BHV-1 in criollos bovines from Parinacochas Province, with a prevalence superior to described in dairy herds of the country.

Thesis (Ph. D.)--University of Missouri--Columbia, 2002. / Typescript. Vita. Includes bibliographical references (leaves 89-103). Also available on the Internet.

Viruses are obligate parasites that use the host cellular machinery to produce progeny virions. The host responds to this invading pathogen by induction of the immune system; however, the virus employs a variety of strategies to overcome these attacks. The complexity of the virus-host interaction is of great interest to researchers with aims to both characterize the relationship and target steps of the viral life cycle to hinder infection. Many targeted tactics employ single protein analysis; however, approaches that examine the whole set of virus/host interactions are available. Transcriptional alterations within host cells have been determined for many virus- host interactions by micro-array techniques; however little is known about the effects on cellular proteins. This study uses a quantitative mass spectrometric-based method, SILAC, to study differences in a host cell's proteome with infection by a virus. Mammalian reoviruses and herpes simplex viruses are prototypical viruses commonly studied to determine virus life cycle and interactions with hosts. Using three strains of reoviruses and one HSV1 strain, cells were infected to identify differentially regulated proteins at different times. Thousands of proteins were identified for each virus type, some up or down regulated after infection. Biological functions and network analyses were performed using online networking tools. These pathway analyses indicated numerous processes including cell death and inflammatory response are affected by T1L reovirus infection. Comparing reovirus strains revealed a greater overall proteomic change in host function when infected with the more pathogenic T3DC strain. For the HSV infection, host proteins altered during the different immediate early, earlyand late phases of infection helped characterize the host-virus interaction parallel to the virus life cycle. Overall, my study has characterized proteomic changes in different virus infection systems, identifying numerous novel cellular functional pathways and specific proteins altered during virus infections, specifically the secretogranin II protein that had opposite types of regulation in reoviruses and HSV and was examined for its effects on virus replication. Further studies on the novel proteomic characteristics may provide greater understanding to the complex virus-host interactome, leading to possible antiviral targets. / October 2015

No description available.