C Morin

Two 19-base-long oligonucleotides were synthesized, one complementary to the normal human beta-globin gene (beta A) and one complementary to the sickle cell beta-globin gene (beta S). The nonadecanucleotides were radioactively labeled and used as probes in DNA hybridization. Under appropriate hybridization conditions, these probes can be used to distinguish the beta A gene from the beta S allele. The DNA from individuals homozygous for the normal beta-globin gene (beta A beta A) only hybridized with the beta A specific probe; the DNA from those homozygous for the sickle cell beta-globin gene (beta S beta S) only hybridized with the beta S specific probe. The DNA from heterozygous individuals (beta A beta S) hybridized with both probes. This allele-specific hybridization behavior of oligonucleotides provides a general method for diagnosis of any genetic disease which involves a point mutation in the DNA sequence of a single-copy gene.

We have used oligonucleotide-directed mutagenesis to make a specific change in the beta-lactamase (EC 3.5.2.6) (ampicillin resistance) gene of the plasmid pBR322. Evidence suggests that the active site for this enzyme may include a serine-threonine dyad (residues 70 and 71). By priming in vitro DNA synthesis with a chemically synthesized 16-base oligodeoxyribonucleotide, we have inverted the Ser-Thr dyad to Thr-Ser and thereby generated a mutant with an ampicillin-sensitive phenotype. This "double-mismatch" method is relatively simple and also very general because detection of mutants is at the level of DNA and involves only colony hybridization. Accordingly, the procedure can be applied to any DNA sequence and does not depend on the phenotype of the mutant.

We report on 162 cases of human papillomavirus (HPV) infection of the cervix seen in a two-year period in which the cell sample showed such marked atypia that errors of interpretation could easily have been made. These atypical condylomata are difficult to diagnose cytologically as well as histologically because they mimic dysplasia or carcinoma in situ and, on smears, even invasive squamous carcinoma. HPV particles associated with fibrillar material were found within nuclei of these lesions; their nature was further proved by the immunoperoxidase test. This new form of HPV infection of the cervix showed a 9.1% rate of progression to more advanced cervical lesions. The cytologic finding of atypical condylomata is an indication for colposcopy, confirmative biopsy and appropriate treatment.

OBJECTIVE: To assess the prevalence of genital bacterial infection among women with vulvar vestibulitis syndrome and to evaluate the association of several potential risk factors with the occurrence of the syndrome. METHODS: Fifty-seven women referred for dyspareunia who satisfied Friedrich's criteria and had symptoms for at least 6 months were recruited as cases. Controls included 173 patients without dyspareunia seen at a private clinic. Cases and controls were aged 18-35 years and were not pregnant. RESULTS: Among cases, the prevalences were low for genital infection with gonorrhea (0%), Chlamydia (0%), Trichomonas (0%), Mycoplasma (0%), Gardnerella (14%), and Candida (8.8%). Ureaplasma was detected in the Bartholin glands of ten affected women (17.5%). Human papillomavirus DNA was detected in only three cases (5.3%) based on polymerase chain reaction assays on vestibular biopsies. The relative risk (RR) of the syndrome was related to some aspects of sexual and reproductive history. In particular, the RR in women who had used oral contraceptives (OCs) early (before age 17) reached 11.0 (95% confidence interval [CI] 1.3-97.1) relative to those who had never used OCs. Women who had first intercourse at age 15 or earlier had a 3.3-fold increase in RR (95% CI 1.4-8.0) compared to those who had first intercourse at age 16 or later. CONCLUSION: Our data provide little support for the idea that infection causes the vulvar vestibulitis syndrome. Hormonal factors such as early OC use may be involved in the etiology of this condition.

The peroxidase-antiperoxidase technique with the use of paraffin sections of 67 cervical biopsy specimens and an antiserum cross-reactive with all papillomaviruses provided immunologic confirmation for the observation that papillomavirus infection of the cervix is not uncommon and that it most often presents as a flat, colposcopically unremarkable lesion. Papillomavirus antigen was detected in 21 or 35 condylomata of the cervix. Antigen-positive nuclei were found in the upper layers of the epithelium. Electron-microscopic examination of five reprocessed antigen-positive sections revealed, in each instance, papillomavirus particles in the nuclei of the most superficial layers of the condylomatous epithelium. The viral antigen was not detected in dysplasia, carcinoma in situ, or invasive carcinoma.