Detection and typing of human papilloma virus DNAs in normal cervix, intraepithelial neoplasia and cervical cancer in Bangkok.

We detected and typed HPV-DNA by polymerase chain reaction (PCR) in cervico-vaginal lavages of 102 women with normal cervical cytology, 57 patients with cervical intraepithelial neoplasia (CIN), and 23 cervical cancer patients. HPV-DNA detection and typing by in situ hybridization were also performed in cervical biopsies from CIN lesions and cancers. Five percent of women with normal cervical cytology, 46% of CIN, and 61% of cervical cancer were positive for HPV-DNA. Of CIN cases with positive HPV-DNA, 69, 15, 8, 4 and 4% were HPV-16, -33, -18, -11 and -16/33 respectively. Of cervical cancer cases with positive HPV-DNA, 86% were HPV-16, 7% were HPV-16/33, 7% were HPV-18/31. HPV typing was performed in biopsies from 37 CIN and 18 cervical cancers by in situ hybridization. By this method, 38% of CIN were HPV-DNA positive, of which 71% were HPV-16 and 7% were each of HPV-11, -18, -31 and -33. Thirty-nine percent of cervical cancers were positive, of which 71% and 29% were HPV-16 and HPV-16/18 respectively.

Genotypic and phenotypic implications in paroxysmal nocturnal hemoglobinuria (PNH): a preliminary investigation.

The genetic and biochemical defects underlying paroxysmal nocturnal hemoglobinuria (PNH) have recently been elucidated. The deficiency of the surface expression of glycosylphosphatidylinositol (GPI)-anchored proteins caused by a somatic mutation of the PIG-A gene, an X-chromosomal gene that participates in the first step of the GPI anchor synthesis, has been shown to be responsible for PNH in all patients. The mutations of PIG-A studied to date are highly heterogeneous. They are however mainly of the frameshift type (61.5%). The characteristic abnormalities of PNH phenotypes has also been shown especially by DAF- and/or CD59-based fluorescent immunocytometry. A great degree of heterogeneity in the patterns and levels of expression of GPI-anchored proteins in various cell types was demonstrated indicating a discrepancy of lineage involvement. In this investigation, major blood cell populations, i.e erythrocytes and granulocytes were analyzed immunophenotypically, the mutations of PIG-A were identified by heteroduplex analysis and nucleotide sequencing and the consequences of PIG-A mutations were observed. All the mutations identified in 9 patients with PNH resulted in complete loss of function as clones of affected granulocytes completely negative for CD59 expression were shown in all patients. Interestingly, granulocytes in these patients contained variable proportions of affected cells varied from 50% to 100% and four of the patients had erythrocytes with diminished expression of GPI-anchored DAF and CD59 coexisting with normal and completely negative cells. Immunophenotypic analysis of reticulocytes in peripheral blood of patients with PNH demonstrated the conserved patterns of DAF and CD59 expression in circulating erythroid cells and the discrepancies between granulocytic and erythroid lineages. These findings suggested that the characteristics of abnormal phenotypes which appear to be highly variable between different hematopoietic lineages are not solely caused by mutation of PIG-A but are influenced by other factor(s).