Antimicrobial peptide response to group B Streptococcus in human extraplacental membranes in culture.

OBJECTIVE: Streptococcus agalactiae (GBS) is an important cause of chorioamnionitis. This study characterizes GBS colonization and stimulation of antimicrobial responses in human extraplacental membranes using an ex vivo transwell two-compartment system of full-thickness membranes and live GBS. STUDY DESIGN: Human extraplacental membranes were affixed to transwell frames (without synthetic membranes). Live GBS was added to the decidual side of membranes in transwell cultures, and cocultures were incubated for 4, 8 and 24 h. GBS recovery from homogenized membranes and culture medium was determined by enumerating colony forming units (CFU) on blood agar. Antimicrobial peptide expression was identified using immunohistochemistry and ELISA. GBS killing by HBDs was assessed in vitro by incubating GBS with different human beta defensins (HBDs) for 3 h, then enumerating CFU. RESULTS: GBS recovery from membranes markedly decreased over time (P < 0.05). The antimicrobial peptides HBD-1, HBD-2, HBD-3, and lactoferrin were expressed in both GBS-exposed and non-exposed tissues. Notably, a pattern of localized increased HBD-2 in the amnion of GBS-infected tissue was observed. Moreover, GBS-treated membranes released increased amounts of HBD-2 into the amniotic and decidual compartments of the transwell cultures after 24 h (P < 0.05). In bacterial cultures, HBD-2 decreased GBS viability in a concentration-dependent manner (P < 0.05). CONCLUSION: Innate immune responses in ex vivo human extraplacental membranes suppress GBS growth. HBD-2 was implicated in this GBS suppression with evidence of signal transduction across the tissue. Antimicrobial peptides may be important for innate immune defense against intrauterine GBS infections during pregnancy.

Comparison of microscopic examination and human papillomavirus DNA subtyping in vulvar lesions of premenarchal girls.

STUDY OBJECTIVE: The purpose of this study is to compare the microscopic examination and human papillomavirus (HPV) DNA subtyping of vulvar specimens from premenarchal girls clinically diagnosed with condyloma to determine whether DNA subtyping aids in the diagnostic process. DESIGN: A retrospective chart review was performed on all premenarchal girls who underwent surgical treatment of clinically diagnosed condyloma between 1993 and 1999 at the University of Michigan Medical Center by the Pediatric and Adolescent Gynecology Service. Tissue was sent for pathologic evaluation and in 10 patients the specimens also underwent DNA subtyping. One patient had prior DNA subtyping. All the other lesions were surgically ablated. The microscopic slides were reviewed by a single pathologist blinded to the study. SETTING: The study was performed in a tertiary care university hospital. PARTICIPANTS: The study group included 11 premenarchal girls with an average age of 2.3 yr. MAIN OUTCOME MEASURES: The charts were reviewed for previous HPV treatment, maternal history of HPV, history of sexual abuse, microscopic diagnosis, and HPV DNA subtyping. RESULTS: Four patients had prior surgical treatment and two patients had undergone prior medical treatment. The microscopic diagnosis was condyloma in 8 patients, chronic dermatitis in 2 patients, and 1 patient had VIN 2-3. All 11 specimens tested positive for HPV DNA, 10 specimens contained at least one of the low-risk subtypes (6, 11, 42, 43, 44), and 1 tested positive for low-risk as well as intermediate/high-risk HPV subtypes (16, 18, 31, 33, 35, 45, 51, 52, 56). CONCLUSIONS: Although all the patients with a clinical diagnosis of condyloma tested positive for HPV DNA, only 9 of 11 were definitely diagnosed with HPV-related pathology by microscopic examination. Therefore, in premenarchal patients with verrucous lesions in the anogenital area, microscopic evaluation alone may be inadequate as a confirmatory test when a positive clinical diagnosis has been made, and HPV DNA subtyping should be considered to avoid confusion with the diagnosis.

Colposcopy in pregnancy: directed brush cytology compared with cervical biopsy.

OBJECTIVE: To evaluate colposcopically directed brush cytology as a substitute for directed biopsy of acetowhite lesions identified during pregnancy. METHODS: Pregnant patients eligible for the study were referred for colposcopic evaluation for either newly diagnosed abnormal cervical cytology or follow-up of a previously diagnosed squamous intraepithelial lesion (SIL). All patients with acetowhite lesions underwent colposcopically directed brush cytology followed by directed biopsy. RESULTS: Of 81 pregnant patients referred, 50 paired samples of colposcopically directed brush cytology and directed biopsies were evaluated from 49 patients. One patient was sampled in the first and third trimesters and one patient's brush cytology was unsatisfactory for interpretation because of clumping artifact, leaving 49 brush-biopsy pairs that were satisfactory for examination. One patient in the study group had an intrauterine fetal death of uncertain cause, remote from the time of biopsy. Compared with the corresponding biopsy, the directed brush caused significantly less blood loss (P < .001). For all diagnostic categories, directed cytology demonstrated a good degree of correlation with biopsy (kappa = 0.73). The brush technique correctly identified 12 of 14 cases (86%) of biopsy-proved cervical intraepithelial neoplasia II-III as high-grade SIL. If one considers "atypical squamous cells, favor human papillomavirus effect" as a true positive, brush sensitivity was 88 +/- 9% and specificity was 74 +/- 12%, with an accuracy of 80%. CONCLUSION: In the absence of lesions suspicious for carcinoma, colposcopically directed brush cytology is a safe substitute for directed biopsy in pregnant patients.

An analysis of two versus three grades for endometrial carcinoma.

Introduction. The current grading of uterine endometrioid adenocarcinoma utilizes a three-grade system based on the amount of nonsquamous solid histologic architecture. Of these three grades, we questioned the practical clinical utility of the intermediate grade. Methods. We retrospectively reviewed endometrial biopsy and uterine histology specimens, quantifying the percentage amount of nonsquamous solid tumor by intervals of 10. We then compared these percentage values to other histopathologic prognostic variables. Results. Eighty-five Stage I and II endometrioid adenocarcinoma patients had their preoperative endometrial curettings and operative hysterectomy pathology reviewed independently by two gynecologic pathologists for surgical staging and outcome with a mean follow-up of 6 years. Using a two-tiered system for assessing uterine tumor grade with a delineating value of 20% nonsquamous solid tumor, we found less interobserver variation (kappa = 0.966) compared to the current three-tiered grading system (kappa = 0.526). There were no differences between the two- and three-tiered grading systems regarding myometrial invasion, lymph vascular space invasion, and survival. In the diagnosis of endometrial biopsies, the two-tiered system also improved the prediction of uterine histology grade over the three-tiered system, 90 and 63%, respectively. Conclusion. A two-grade architecture system with a delineation value of 20% would be more reliable and less cumbersome and would have the same or better prognostic significance as the currently used three-grade system.

The diagnosis of gonorrhea in a low-prevalence female population: enzyme immunoassay versus culture.

In a low-prevalence population for sexually transmitted disease, accurate methods of identifying Neisseria gonorrhoeae infections are of paramount importance. Tests with low sensitivity are of concern to each practitioner. Of no less concern, however, are the possible social and emotional consequences of a false-positive test in a family-based, rural, or military community. Several centers serving high-prevalence populations have shown that the Gonozyme Enzyme Immunoassay may provide many advantages in the rapid detection of gonococcal infection in these groups. Few studies, however, have evaluated its efficacy in low-prevalence populations. We compared the Gonozyme with Martin-Lewis culture medium in 1490 women considered at risk for infection with N gonorrhoeae at Keesler USAF Medical Center, Mississippi. Fifty-three women had culture-proved infections (3.6% disease prevalence). The sensitivity and specificity of the Gonozyme in this population were 92.4 and 97.2%, respectively. However, the predictive value of a positive test was only 55.1%, mandating culture confirmation. It appears that in a low-prevalence female population, standard culture techniques provide a more efficient and reliable means of detecting N gonorrhoeae than does the Gonozyme Enzyme Immunoassay.

Hemodynamic predictors of myocardial ischemia during halothane anesthesia for coronary-artery revascularization.

The authors undertook a prospective study of 30 patients undergoing halothane anesthesia for coronary-artery revascularization to ascertain which clinically monitored hemodynamic variables--or combination of variables--associated with myocardial oxygen supply and demand best predict myocardial ischemia. Simultaneous recordings of electrocardiogram (lead II and V5), systemic, central venous, pulmonary artery, and pulmonary artery occluded pressures were analyzed for correlation with ischemic episodes. Ischemia occurred with significant increases (P less than 0.0001) in heart rate, central venous pressure, and pulmonary artery occlusion pressure and with significant decreases (P less than 0.0001) in systolic and mean arterial blood pressure and in coronary perfusion pressure (mean arterial minus pulmonary artery occluded pressure). There was no correlation between ischemia and either hypertension (systolic blood pressures up to 200 mmHg) or the rate-pressure product. Systemic systolic blood pressure, systemic mean arterial blood pressure, and coronary perfusion pressure as single determinants were the most useful to monitor in avoiding myocardial ischemia. A combination of systemic arterial blood pressure (systolic or mean) and filling pressure (central venous or pulmonary artery occluded) was generally as useful but not more so than the preceding single variables in avoiding ischemia. Rate-pressure product was not of value in this regard. Patients were divided into three groups according to preoperative left ventricular (LV) function to determine whether pulmonary artery occluded pressure (PAOP) was more useful than central venous pressure (CVP) as either a predictor of ischemia or an index of cardiac filling: normal LV function (Group I), moderately abnormal LV function (Group II), and markedly abnormal LV function (Group III). PAOP offered no advantage over CVP for either purpose, except in some Group III patients.