Structural and kinetic analyses of herpes simplex virus type 1 latency-associated transcripts in human trigeminal ganglia and in cell culture.

Only one herpes simplex virus type 1 (HSV-1) gene is expressed in sensory neurons of latently infected animals and humans, yielding two RNAs, called latency-associated transcripts (LATs). The LATs appear to modulate virus reactivation. In mice and rabbits the 5' origins, kinetics of synthesis, and splicing pattern of the LATs are well established. Because these details of LAT structure and expression have not been defined in humans, we sought to do so. Using primer extension and Northern hybridization analyses, we demonstrate that in human trigeminal ganglia, the smaller (1.35 kb) HSV-1 transcript differs from the larger (1.85 kb) LAT by excision of an intron near its 5' end; they are otherwise colinear, and 5' coterminal. In infected cells only the 1.85 kb LAT is detected. Its expression is inhibited by cycloheximide or acyclovir, indicating this LAT is synthesized late in the viral replicative cycle. All of these features of the LATs in humans are consistent with those reported in rabbits and mice and further validate the animal models of human HSV-1 infection.

Herpes simplex virus type 1 transcription is not detectable in quiescent human stromal keratitis by in situ hybridization.

PURPOSE: The purpose of this study was to determine whether herpes simplex virus (HSV) transcripts are present in the corneas of patients with chronic herpetic stromal keratitis. METHODS: Corneal buttons from patients with a history of stromal keratitis, but no ongoing active disease, together with positive and negative control tissues, were analyzed by in situ hybridization using single-stranded RNA probes for all three classes of viral lytic cycle transcripts as well as for the latency-associated transcripts (LATs). Tissues also were screened for presence of HSV genomic DNA using the polymerase chain reaction (PCR). RESULTS: HSV DNA was detected in 7 of 13 quiescent corneas by PCR, but no viral transcripts were detected in any of these corneas by in situ hybridization. CONCLUSIONS: At the level of detection afforded by in situ hybridization, HSV persistent in scarred human corneas after stromal keratitis appears to be transcriptionally dormant. This contrasts with the situation in neurons of latently infected sensory ganglia, in which LATs are present at high levels.

Directionality and further mapping of varicella zoster virus transcripts.

Our laboratory previously identified and preliminarily mapped 58 viral RNA transcripts in varicella zoster virus (VZV) infected cells (Ostrove et al., 1985). This study was initiated to more precisely map these transcripts, to identify additional transcripts, and to determine transcript directionality. To accomplish this, 32 overlapping BamHI, EcoRI, and SmaI fragments representing 99.7% of the genome were cloned into pGEM-2, a plasmid which contains a multiple cloning site flanked by SP6 and T7 RNA polymerase promoters. Each of these clones was used to produce 32P-labeled double-stranded DNA probes to detect transcripts homologous to either strand of the VZV insert, and single-stranded [32P]RNA probes in order to detect RNAs of either polarity. These probes were hybridized to Northern blots of VZV-infected cell RNA. In all, 77 RNAs were detected with both DNA and RNA probes. The direction of transcription and localization of 57 of the 58 previously identified RNAs and of 20 newly recognised abundant transcripts were determined. Thirty-three additional low-abundance transcripts were detected only by the relatively more sensitive RNA probes. A map indicating the directionality and approximate locations of the abundant VZV transcripts was constructed.

Socioeconomic status and health: what we know and what we don't.

In the past 15 years, we have seen a marked increase in research on socioeconomic status (SES) and health. Research in the first part of this era examined the nature of the relationship of SES and health, revealing a graded association; SES is important to health not only for those in poverty, but at all levels of SES. On average, the more advantaged individuals are, the better their health. In this paper we examine the data regarding the SES-health gradient, addressing causal direction, generalizability across populations and diseases, and associations with health for different indicators of SES. In the most recent era, researchers are increasingly exploring the mechanisms by which SES exerts an influence on health. There are multiple pathways by which SES determines health; a comprehensive analysis must include macroeconomic contexts and social factors as well as more immediate social environments, individual psychological and behavioral factors, and biological predispositions and processes.

Development of an improved product enhanced reverse transcriptase assay.

A PCR based reverse transcriptase (RT) assay was developed that has 10(4)-fold higher sensitivity than conventional nucleotide incorporation assays and allows discrimination between false positive results generated by cellular polymerases and positives resulting from authentic RT activity. Recently, several reverse transcriptase (RT) assays have been developed where a reverse transcriptase reaction is performed on an RNA template/DNA primer combination. A specific region of the cDNA product is then amplified by the polymerase chain reaction to increase the sensitivity of cDNA detection. These reverse transcriptase assays up to 10(6)-fold more sensitive at detecting retroviruses than conventional methods. The drawback to these assays with increased sensitivity is the increased incidence of false positive results generated by cellular polymerases that can reverse transcribe. The MS2 bacteriophage RNA template and primers from one of the recently developed assays were used as the basis to develop the assay. A simple high resolution agarose gel was used as the endpoint for the assay without compromising sensitivity. In addition, the pH of the RT reaction was lowered to pH 5.5, the RT incubation was 1 h, and protease inhibitors were added to the RT reaction components. These modifications yield an assay that can discriminate between authentic RT activity and contaminating cellular polymerases.

Objective and subjective assessments of socioeconomic status and their relationship to self-rated health in an ethnically diverse sample of pregnant women.

A new measure of subjective socioeconomic status (SES) was examined in relation to self-rated physical health in pregnant women. Except among African Americans, subjective SES was significantly related to education, household income, and occupation. Subjective SES was significantly related to self-rated health among all groups. In multiple regression analyses, subjective SES was a significant predictor of self-rated health after the effects of objective indicators were accounted for among White and Chinese American women; among African American women and Latinas, household income was the only significant predictor of self-rated health. After accounting for the effects of subjective SES on health, objective indicators made no additional contribution to explaining health among White and Chinese American women; household income continued to predict health after accounting for subjective SES among Latinas and African American women.

Psychological predictors of good health in three longitudinal samples of educated midlife women.

Concurrent and longitudinal associations between cognitive and affective personality variables--intellectual efficiency (IE), anxiety, and hostility--and observer ratings of physical health were examined in 3 longitudinal samples of women: Mills Longitudinal Study (n = 101); Radcliffe Study (RS, n = 118); and University of California, San Francisco Study (n = 44). Observer ratings of health were based on participants' reports of health problems. The California Psychological Inventory (H. G. Gough, 1996) IE, Hostility, and Anxiety Scales were used in all studies at Times 1 and 2, except in RS, when at Time 1 the Zung Anxiety (W. K. Zung, 1971) and the Profile of Mood States (D. M. McNair, M. Lorr, & L. F. Droppleman, 1971) Hostility Scales were used. In the majority of analyses, IE was positively associated with good health, and Anxiety and Hostility were negatively associated with health. IE was the strongest independent predictor of health, indicating that cognitive characteristics may have an important role in health and should be examined further.

Predicting women's well-being in midlife: the importance of personality development and social role involvements.

Theories of adult development suggest that both personality and social roles are sources of adult well-being, but most research has examined only social roles. An integrated model was used, including personality, number of roles, and role quality, to predict well-being in 2 longitudinal studies of college-educated women. Results for both samples indicated that role quality and personality development were important components of the path to well-being, whereas number of roles, occupied was important mainly in early adulthood. Moreover, the results provided support for E. Erikson's (1968) notion of the importance of the sequencing of personality development for later well-being. Path analyses indicated that engagement in multiple roles during early adulthood facilitated the development of identity, which predicted generativity and role quality, which in turn predicted well-being.

Women's personality in middle age. Gender, history, and midcourse corrections.

This article examines several key features of the course of adult development in the cohort of women born during the baby boom. By focusing on the women in this group and comparing their experience with that of older cohorts and research on men, the authors demonstrate the need for models of aging that take account of the inter-sections of history, gender, and individual development. Concepts proposed as universal features of middle age (midlife crisis, generativity, aging), as well as those proposed as specific to women (empty nest, menopause) are examined. Perhaps most important, certain features not commonly viewed as particularly important in women's middle aging (midlife review, identity, confident power) are shown to be central. The need for further research examining these same processes among men and different groups of women is underscored.

The Relationship of Socio-economic Status, Labor Force Participation, and Health among Men and Women.

Using survey data from an extensive sample of Californians in the United States, we examined the relationship among indicators of socio-economic status (SES) and health for men and women in and out of the paid labor force. In contrast to data reported from the mid-1980s in which correlations among SES variables were stronger among men than among women, correlations among education, income, and occupation were similarly high among women and men. All SES variables were significantly related to health, such that as SES increased, self- reported health status increased. Multiple regression analyses demonstrated an additive effect of labor force participation and SES in predicting health among men, with main effects both of labor force participation and of income and education on health. Among women, there was an interactive effect of these variables, such that the relationship of income and of education to health was more pronounced among women who were not in the paid labor force.

Relations among Socioeconomic Status Indicators and Health for African-Americans and Whites.

This investigation explored the relationship of socioeconomic status (SES) to physical and mental health in two nationally representative samples of whites and African-Americans. We examined the interrelations among SES variables and assessed their contribution to health for the two racial groups. Throughout, we assessed the contribution of a less traditional indicator of SES-wealth-in the SES-health relationship. As we expected, African-Americans had lower levels of education, household income, and wealth than whites. Unexpectedly, however, the strength of the interrelationships among the three SES indicators did not differ for African-Americans and whites. In addition, we found that SES operated to affect health in a very similar fashion for African-Americans and whites. We found that wealth, in addition to more traditional indicators of SES (education and household income), made a unique and significant contribution to explaining both physical and mental health. Examining relations of different SES indicators to health across groups is critical to eliminating persistent social inequalities in health.

Differential regulation by varicella-zoster virus (VZV) and herpes simplex virus type-1 trans-activating genes.

Transient expression assays were performed in Vero cells in order to compare varicella-zoster virus (VZV)-encoded trans-activating proteins [defined by the products of open reading frames (ORF) 4 and 62] with herpes simplex virus type-1 (HSV-1) trans-activating proteins, ICP4 and ICP0, with respect to activation of gene expression. We demonstrate that the product of VZV ORF4 and ORF62 (which are the HSV-1 analogs of ICP27 and ICP4, respectively) stimulate a variety of viral and cellular gene promoters, including the HSV-1 thymidine kinase (tk) promoter. On the other hand, expression of a recombinant vector containing the VZV tk promoter could not be stimulated, by HSV-1 infection or by the HSV-1 ICP4 or ICP0 proteins expressed during cotransfection experiments. These data suggest different mechanisms of activation of the VZV and the HSV-1 tk gene promoters by "trans-activating" factors.

Resistance of viruses to antiviral drugs.

In recent years, substantial advances in antiviral therapy have been made; however, in the immunocompromised host, antiviral drug resistance is becoming an issue of increasing clinical importance. Understanding the mechanism of action of antiviral agents, especially those used to treat herpesvirus infections, may enable us to design new therapeutic agents and better treatment regimens to deal with antiviral drug resistance.

Characterization of a potent varicella-zoster virus-encoded trans-repressor.

Using a transient expression assay in Vero cells, we have shown that the protein product from gene 61 of varicella-zoster virus (VZV) can repress the function of the VZV encoded trans-activators on putative viral immediate-early, early, and late gene promoters. The repression is exerted at the transcriptional level and requires functional gene 61 protein. This trans-repressor is the herpes simplex type 1 ICP0 (a trans-activator) homolog, as defined by gene location, the sharing of a cysteine-rich putative zinc-binding finger in the amino-terminal region, and limited amino acid homology. Open reading frame 61 (ORF61)-mediated trans-repression appears to be specific for VZV-encoded trans-activators in that it has no effect on simian virus 40 and Rous sarcoma virus promoters. Moreover, it does not inhibit trans-activation of the human T-lymphotropic virus type I and human immunodeficiency virus long terminal repeats by tax and tat genes, respectively. We constructed plasmids with mutations in ORF61 and tested them for their ability to inhibit trans-activator (VZV genes 4 and 62)-mediated activation of the viral thymidine kinase promoter-chloramphenicol acetyltransferase construct. Mutants containing interruptions in ORF61 lost their trans-repressing ability, as demonstrated at both the protein and steady-state RNA levels. These results suggest that the ORF61 protein product can mediate down-regulation of VZV gene expression.

Mapping of two varicella-zoster virus-encoded genes that activate the expression of viral early and late genes.

A transient assay system was used to identify varicella-zoster virus (VZV)-encoded genes whose products are able to activate the expression of an early gene promoter, the thymidine kinase (tk) promoter, and a late gene promoter, and the glycoprotein I (gpI) promoter. Vero cells were cotransfected with individual cloned DNA fragments spanning the entire VZV genome and with the recombinant construct p1tkCAT which contained the chloramphenicol acetyl transferase (CAT) gene under the control of putative regulatory sequences. Five- to 20-fold increases in the expression p1tkCAT was observed in cotransfections with plasmids containing VZV open reading frame (ORF)4 (map location 0.02-0.03) or ORF62 (0.82-0.86). Expression of p68CAT (contains -682 to +222 bp relative to the AUG of gpI) was also enhanced by the products of ORF4 and ORF62. Synergy between ORF4 and ORF62 products was observed in the activation of p68CAT, resulting in a 22-fold increase in CAT activity. RNA analysis indicated that activation of these promoters was at the transcriptional level. A VZV-encoded "repressor" sequence, containing ORF60 and ORF61, was also identified which repressed expression of p1tkCAT and modulated its activation by ORF4 and ORF62.

Beyond scripts: a note on the capacity of right hemisphere-damaged patients to process social and emotional content.

Individuals with right hemisphere damage (RHD) can comprehend simple scripts but have difficulties understanding more complex forms of discourse such as stories and jokes. This study sought to discover the point beyond simple scripts at which such patients begin to experience difficulties. Three-sentence, script-like vignettes which described everyday situations were created along two major dimensions: a social dimension (reflecting presence or absence of interactions between characters); and an emotional dimension (reflecting a positive, negative, or neutral emotion felt by a protagonist in the situation). Right hemisphere-damaged patients were asked to provide emotional characterizations of, and continuations to, these situational vignettes. Results indicated that individuals with right hemisphere damage were more likely than control subjects to attribute a positive emotional state to a character in a neutral situation. In other respects, there were no significant differences between RHD subjects and controls, although the RHD subjects made significantly more factual errors in their continuations than did control subjects.

Varicella-zoster virus complements herpes simplex virus type 1 temperature-sensitive mutants.

Varicella-zoster virus (VZV) can complement temperature-sensitive mutants of herpes simplex virus. Of seven mutants tested, two, carrying mutations in the immediate-early ICP4 and ICP27 proteins, were complemented. This complementation was not seen in coinfections with adenovirus type 5 or cytomegalovirus. Following transfection into CV-1 cells, a DNA fragment containing the VZV short repeat sequence complemented the ICP4 mutant. These data demonstrate a functional relationship between VZV and herpes simplex virus and have allowed localization of a putative VZV immediate-early gene.