The association of HSV 1 and 2 with atherosclerosis defined by CRP level

A study of the association of Herpes simplex virus 1 and 2 exposure to early atherosclerosis using high C-reactive protein level as a marker was carried out in US born, non-pregnant, 20-49 year olds participating in a national survey between 1999 and 2004. Participants were required to have valid results for Herpes simplex virus 1 and 2 and C-Reactive Protein for inclusion. Cases were those found to have a high C-reactive protein level of 0.3-1 mg/dL, while controls had low to normal values (0.01-0.29 mg/dL). Overall, there were 1211 cases and 2870 controls. Mexican American and non-Hispanic black women were much more likely to fall into the high cardiac risk group than the other sex race groups with proportions of 44% and 39%, respectively. ^ Herpesvirus exposure was categorized such that Herpes simplex virus 1 and 2 exposure could be studied simultaneously within the same individual and models. The HSV 1+, HSV 2- category included the highest percentage (45.63%) of participants, followed by HSV 1-, HSV 2- (30.16%); HSV 1+, HSV 2+ (15.09%); and HSV 1-, HSV 2+ (9.12%) respectively. The proportion of participants in the HSV 1+, HSV 2- category was substantially higher in Mexican Americans (63%-66%). Further, the proportion in the HSV 1+, HSV 2+ category was notably higher in the non-Hispanic black participants (23%-44%). Non-Hispanic black women also had the highest percentage of HSV 1-, HSV 2+ exposure of all the sex race groups at 17%. ^ Overall, the unadjusted odds ratios for atherosclerotic disease defined by C-reactive protein with HSV 1-, HSV 2- as the referent group was 1.62 (95% CI 1.23-2.14) for HSV 1 +, HSV 2+; 1.3 (95% CI 1.10-1.69 for HSV 1+, HSV 2-; and 1.52 (95% CI 1.14-2.01). When the study was stratified into sex-race groups, only HSV 1+, HSV 2- in the Non-Hispanic white men remained significant (OR=1.6; 95% CI 1.06-2.43). Adjustment for selected covariates was made in the multivariate model for both the overall and sex-race stratified studies. High C-reactive protein values were not associated with any of the Herpesvirus exposure levels in either the overall or stratified analyses. ^

Regulation of apoptosis during the pathogenesis of non -melanoma skin cancer

Previous studies from our lab have shown distinctive patterns of expression of bcl-2 gene family members in human nonmelanoma skin cancer (NMSC). To further evaluate the significance of these observations and to study the effects of cell death deregulation during skin carcinogenesis, we generated a transgenic mouse model (HK1.bcl-2) using the human keratin 1 promoter to target the expression of a human bcl-2 minigene to the epidermis. Transgenic protein expression was confirmed in all the layers of the epidermis except the stratum corneum using immunohistochemistry. Multifocal epidermal hyperplasia, without associated hyperkeratosis, was observed in newborn HK1.bcl-2 mice. Immunofluorescence staining using monoclonal antibodies specific for a variety of differentiation markers revealed aberrant expression of keratin 6 (K6) in the transgenic epidermis. Epidermal proliferative indexes, assessed by anti-BrdUrd immunofluorescence staining, were similar in control and transgenic newborn mice, but suprabasal proliferating cells were seen within the hyperplastic areas of the transgenic mouse skin. Spontaneous apoptotic indices of the epidermis were similar in both control and HK1.bcl-2 transgenic newborn mice, however, after UV-B irradiation, the number of "sunburn cells" was significantly higher in the control compared to the HK1.bcl-2 transgenic animals.^ Adult HK1.bcl-2 and control littermate mice were used in UV-B and chemical carcinogenesis protocols including DMBA + TPA. UV-B irradiated control and HK1.bcl-2 mice had comparable incidence of tumors than the controls, but the mean latency period was significantly shorter in the HK1.bcl-2 transgenic. Both control and transgenic animals included in chemical carcinogenesis protocols required application of both the initiating (DMBA) and promoting (TPA) agents to develop tumors. The frequency, number, and latency of tumor formation was similar in both groups of animals, however, HK1.bcl-2 mice exhibited a rate of conversion from benign papilloma to carcinoma 2.5 times greater than controls.^ Similar carcinogenesis experiments were performed using newborn mice. HK1.bcl-2 mice treated with UV-B plus TPA have a three fold greater incidence of tumor formation compared to controls littermates. HK1.bcl-2 transgenic animals also exhibited a shorter latency for papilloma formation when treated with DMBA plus TPA.^ HK1.bcl-2/v-Ha-ras double transgenic mice shared phenotypic features of both HK1.v-Ha-ras and HK1.bcl-2 transgenic mice, and exhibited focal areas of augmented hyperplasia. These double transgenic mice were susceptible to tumor formation by treatment with TPA alone.^ Cultures of primary keratinocytes were established from control, HK1.bcl-2, HK1.Ha-ras, and HK1.bcl-2/v-Ha-ras newborn mice. Cell viability was determined after exposure of the cells to UV-B irradiation, DMBA, TPA, or TGF-$\beta$1. Internucleosomal DNA fragmentation ("ladders") and morphological cellular changes compatible with apoptotic cell death were observed after the application of all these agents. HK1.bcl-2 keratinocytes were resistant to cell death induction by all of these agents except TGF-$\beta$1. HK1.Ha-ras cells had a higher spontaneous rate of cell death which could be compensated by co-expression of bcl-2.^ These findings suggest that bcl-2 dependent cell death suppression may be an important component of multistep skin carcinogenesis. ^