The environmental cofactors in carcinogenesis in high risk HPV/HIV-positive women

The objective of the present study was to assess the presence of human papilloma virus (HPV) in HIV-infected women, with comparison between the Papanicolaou cytologic technique and the molecular PCR technique, as well as to determine the type of HPV, to measure cellular immunocompetence and to identify the presence of risk factors for the acquisition of HPV infection. Thirty HIV-infected women were selected. Vaginal and endocervical samples were collected from 27 of them. The smears were examined by 3 experienced cytologists to diagnose the presence of HPV by the Papanicolaou technique and the results were compared to HPV detection and typing by PCR. HPV-infected patients were interviewed in order to identify the presence of risk factors for the acquisition of the virus. Eight of the 27 patients analyzed (29%) presented HPV in endocervical samples submitted to PCR, 6 of them (75%) presented HPV involving a high risk of development of cervical cancer. For 5 of these patients, the cytologic diagnosis was not confirmed by PCR. When cellular immunocompetence was related to HPV infection, PCR revealed a diagnosis of HPV in 37.50% of the patients at intermediate risk for HPV infection and in 83.33% of the patients at high risk for HPV infection. These immunologically compromised HPV-infected patients are at higher risk of developing cervical neoplasia. We showed here that PCR is adequate for HPV detection and that, if only the Papanicolaou method is used for the follow-up of these patients, we will not provide good prevention of cervical cancer

Effect of isoflavone extracts from Glycine max on human endothelial cell damage and on nitric oxide production

Objective: In this study, we determined the protective effect of isoflavones from Glycine max on human umbilical vein endothelial cell (ECV304) damage induced by hydrogen peroxide (H(2)O(2)) and on nitric oxide (NO) production. Methods: We studied the regulation of NO synthesis in cultured human endothelial cells by phytoestrogens contained in soy extracts in the presence or absence of ICI 182,780 or N(omega)-nitro-L-arginine methyl esther and determined the protective effect of these isoflavones on ECV304 damage induced by H(2)O(2). Results: We show that soy extracts activate NO synthesis in endothelial cells and protect against cell damage. Conclusions: In conclusion, soy isoflavones markedly protect ECV304 cells against H(2)O(2) damage and promote NO synthesizing. Therefore, these isoflavones call potentially act as an NO promoter and as an antioxidant

Efeito da terapia de reposição hormonal sobre o estado férrico

INTRODUÇÃO: Na literatura, numerosas publicações relatam a determinação do estado férrico em crianças, adolescentes e mulheres em fase reprodutiva, no entanto são raras as pesquisas quanto às alterações do ferro em estoque e eritrograma pós-terapia de reposição hormonal (TRH) em pré-menopausadas e menopausadas. O aumento dos estoques de ferro em mulheres pré-menopausadas e menopausadas pode conduzir à elevação do estresse oxidativo e, conseqüentemente, ao risco de desenvolvimento de câncer e doenças cardiovasculares. OBJETIVO: Avaliar o efeito da TRH sobre o eritrograma e o estado férrico em mulheres na pré-menopausa e na menopausa. MÉTODOS: Foram determinados os eritrogramas e as dosagens de ferro, capacidade total de ligação do ferro a transferrina (CTLF) e ferritina séricas em 30 mulheres no climatério antes e após seis meses de TRH com medroxiprogesterona e estradiol. Os eritrogramas, as dosagens de ferro e CTLF foram determinados por meio da utilização de métodos clássicos, e a ferritina, por quimiluminescência. RESULTADOS: Após o uso da TRH, constataram-se significante redução do número de eritrócitos, elevação dos índices hematimétricos e tendência à diminuição nos níveis de ferro sérico e CTLF. Nenhuma alteração significante nos níveis de ferritina e no índice de saturação de transferrina foi detectada após a TRH. DISCUSSÃO E CONCLUSÃO: No presente estudo não foram encontradas alterações nos parâmetros hematimétricos e no estado férrico que impeçam a utilização da TRH no climatério e na menopausa. Os resultados sugerem que a TRH exerceu efeito benéfico sobre o estado férrico nas mulheres no climatério deste estudo, mantendo os estoques de ferro normais e promovendo a elevação dos índices hematimétricos.<br>BACKGROUND: In literature there are many studies about iron status in children, adolescents and fertile women, but investigations of iron stores and erythrocyte parameters variation after hormone replacement therapy (HRT) are rare. Elevated iron stores, oxidative stress, and estrogen deficiency may place premenopausal and menopausal women in a risk of developing heart disease and cancer. OBJECTIVE: To evaluate the effect of HRT in erythrogram and iron status in premenopausal and menopausal women. METHODS: Hematological indices and iron status were assessed by erythrogram, serum ferritin, iron and transferrin iron-binding capacity (TIBC) in 30 pre- and menopausal women before and after HRT with medroxiprogesterone and estradiol. The blood exam, serum iron and iron-binding capacity were determined by laboratory classic methods, while ferritin was measured by quimiluminescent assay. RESULTS: HRT use was followed by a significant reduction in the absolute number of erythrocyte, an increase of hematimetric indexes and a trend towards a reduction of serum iron levels and TIBC. No alterations on serum ferritin and transferrin saturation index were detected after HRT. DISCUSSION AND CONCLUSION: In the present study, alterations in red cell and iron parameters, which could impair the use of HRT in premenopausal and menopausal women, were not observed. Our results suggest that HRT in premenopausal women are beneficial to iron status, maintaining normal iron stores and promoting elevation of red cells indexes

Expression of human protein S100A7 (psoriasin), preparation of antibody and application to human larynx squamous cell carcinoma

Background\ud Up-regulation of S100A7 (Psoriasin), a small calcium-binding protein, is associated with the development of several types of carcinomas, but its function and possibility to serve as a diagnostic or prognostic marker have not been fully defined. In order to prepare antibodies to the protein for immunohistochemical studies we produced the recombinant S100A7 protein in E. coli. mRNA extracted from human tracheal tumor tissue which was amplified by RT-PCR to provide the region coding for the S100A7 gene. The amplified fragment was cloned in the vector pCR2.1-TOPO and sub-cloned in the expression vector pAE. The protein rS100A7 (His-tag) was expressed in E. coli BL21::DE3, purified by affinity chromatography on an Ni-NTA column, recovered in the 2.0 to 3.5 mg/mL range in culture medium, and used to produce a rabbit polyclonal antibody anti-rS100A7 protein. The profile of this polyclonal antibody was evaluated in a tissue microarray.\ud \ud Results\ud The rS100A7 (His-tag) protein was homogeneous by SDS-PAGE and mass spectrometry and was used to produce an anti-recombinant S100A7 (His-tag) rabbit serum (polyclonal antibody anti-rS100A7). The molecular weight of rS100A7 (His-tag) protein determined by linear MALDI-TOF-MS was 12,655.91 Da. The theoretical mass calculated for the nonapeptide attached to the amino terminus is 12,653.26 Da (delta 2.65 Da). Immunostaining with the polyclonal anti-rS100A7 protein generated showed reactivity with little or no background staining in head and neck squamous cell carcinoma cells, detecting S100A7 both in nucleus and cytoplasm. Lower levels of S100A7 were detected in non-neoplastic tissue.\ud \ud Conclusions\ud The polyclonal anti-rS100A7 antibody generated here yielded a good signal-to-noise contrast and should be useful for immunohistochemical detection of S100A7 protein. Its potential use for other epithelial lesions besides human larynx squamous cell carcinoma and non-neoplastic larynx should be explored in future.This research was supported by FAPESP doctoral fellowship n°. 05/50781-2 (Manuela Ramos Barbieri) and grants from CTC/CEPID/FAPESP (grant n°. 1998/14247-6). The authors would like to thank Cristiane A. Pereira and Anemari R. Dinarte for technical assistance, Roberto G. Sanchez for preparation of the figures, Dr. Elettra Greene for translating the Portuguese text, Dr. Clarice Izumi for intellectual advice and assistance and Dalva T. Catto and Silvia E. Bolfarini for secretarial assistance