Host cell tropism of equine herpesviruses: glycoprotein D of EHV-1 enables EHV-4 to infect a non-permissive cell line.

Equine herpesviruses 1 and 4 (EHV-1 and EHV-4) cause equine respiratory disease worldwide. However, only EHV-1 is a cause of abortion and neurological disease, despite the two viruses having all 76 genes in common. In addition EHV-1 has a broader host range in cell culture than EHV-4, as exemplified by the rabbit kidney (RK) cell line that is permissive for EHV-1, but not for EHV-4. Here we describe that when EHV-4 produced in equine cells was inoculated onto RK cells expressing glycoprotein D of EHV-1 (RKgD1), infection developed as clusters of rounded cells, and this infectivity could be passaged in RKgD1 cells. The progeny virus could also infect single RK cells, consistent with EHV-4 acquiring EHV1 gD from the complementing cell line. No such infection was observed for EHV-4 in RK cells expressing EHV-1 glycoprotein C. The results are consistent with gD homologues being major determinants of host cell tropism and raise the possibility that gD may be a factor in the differential pathogenicity of EHV-1 and EHV-4.

In vitro transposon mutagenesis of an equine herpesvirus 1 genome cloned as a bacterial artificial chromosome.

The 150-kbp genome of the alphaherpesvirus equine herpesvirus 1 (EHV-1) strain HVS25A was cloned as a bacterial artificial chromosome (EHV-1 BAC), with mini F plasmid sequences inserted between genes 62 and 63. Transfection of EHV-1 BAC DNA purified from E. coli gave rise to progeny virus that had a similar growth rate and yield in mammalian cell culture to those of parental wild-type EHV-1. Using in vitro mutagenesis with a Mu transposon, a large library of EHV-1 BAC mutants was generated, and sequence analysis indicated that insertions were dispersed randomly across the EHV-1 genome. Following transfections of a pilot sample of mutant EHV-1 BAC DNAs into mammalian cells, no CPE was observable by light microscopy for mutants carrying insertions in genes for the major capsid protein, large tegument protein, glycoprotein K, catalytic subunit of DNA polymerase, or single-stranded DNA-binding protein. Mutants that were able to produce CPE similar to wild-type EHV-1 included those with interruptions in ORFs of several tegument proteins. Analysis of several glycoprotein gene mutants indicated that those carrying insertions near the start of genes encoding glycoproteins E and I were viable, but showed markedly diminished plaque areas. These results were supported by confocal microscopy of transfected or infected cultures. Electron microscopy of cells infected with a gE mutant revealed accumulations of particles within cytoplasmic vesicles, consistent with a partial obstruction of maturation. The transposon library is a resource for comprehensive functional analysis of the HVS25A genome, with multiple mutants available in any of the predicted genes of EHV-1.

Oral misoprostol in the third stage of labor.

OBJECTIVE: To compare the efficacy of intravenous ergometrine, intramuscular oxytocin, and oral misoprostol in the control of postpartum hemorrhage. METHODS: Mean blood loss, rates of blood loss between 500 and 1000 ml, hematocrit fall greater than 10%, and need for additional oxytocic agents and nature and rates of adverse effects were assessed in this prospective, randomized, controlled study. RESULTS: All outcomes were similar in the 3 groups. The main adverse effects in the misoprostol group were temperatures higher than 99 degrees F, which normalized within 2 h and shivering, which was mild and self-limiting. CONCLUSIONS: Oral misoprostol is as effective as conventional oxytocic agents in preventing postpartum hemorrhage and can be recommended for use in low-resource settings.

A predictive model for cesarean section in low risk pregnancies.

OBJECTIVE: A small number of women with low risk pregnancies undergo cesarean section. A model that can predict this risk and therefore identify these women will be of help in several hospitals where personnel and resources are limited. METHODS: The study consisted of 2 parts. All charts of women with low risk singleton pregnancies admitted to labor room over a 5-month period were analyzed. Adjusted odds ratios were calculated to find out relative importance of each risk factor and likelihood ratios were obtained. These were prospectively applied to 1010 consecutive low risk women and the post test probability calculated. Finally the actual incidence of cesarean section was compared with posttest probability derived from predictors. RESULTS: A combination of maternal age >24 years, primiparity and height <150 cm or a combination of any 2 of the 3 variables is significantly associated with increased cesarean section rate. Individually, primiparity, height <150 cm or age >24 years also significantly increased the chances of cesarean section. CONCLUSIONS: A predictive model consisting of maternal age, parity and height can be used to identify low risk pregnant women who are likely to require cesarean section.

Teratoma of the placenta.

Primary non-trophoblastic tumours of the placenta reported to date are chorioangioma and teratoma, both of which are extremely rare. A case of teratoma in a term placenta is reported.

Tamoxifen-induced endometrial changes in postmenopausal women with breast carcinoma.

OBJECTIVES: To assess the effects of tamoxifen (TAM) on the endometrium in postmenopausal women. METHODS: A case control study of postmenopausal women with breast carcinoma, who were undergoing treatment in the Department of Radiotherapy and Surgery at the Christian Medical College Hospital, Vellore, India was done. Thirty-five women who were on tamoxifen (20 mg/day) for a period of at least 6 months formed the study group. Thirty-three women who were not receiving tamoxifen, formed the control group. Subjects in both groups had a pelvic examination and transvaginal sonogram followed by endometrial biopsy. RESULTS: There was a statistically significant difference in the mean endometrial thickness between the study group and control group (7.8+/-6.4 mm vs. 4.0+/-2.0 mm, respectively) More women in the tamoxifen group had an endometrial thickness of >5 mm but the number of women with polyps or hyperplasia of the endometrium did not differ significantly between the two groups. There were no women with endometrial carcinoma in either group. CONCLUSION: All patients on tamoxifen need to be evaluated by clinical examination annually. A transvaginal sonogram and endometrial biopsy/hysteroscopy may be performed on patients with abnormal vaginal bleeding, bloody discharge, staining or spotting.

See & treat protocol for evaluation & management of cervical intraepithelial neoplasia.

BACKGROUND & OBJECTIVES: The incidence of cervical cancer in India remains high, largely due to ineffective screening and poor patient compliance for follow up. A one step procedure where evaluation and treatment are performed at the same sitting may be a good strategy. Therefore the present study was undertaken to evaluate the feasibility of a see and treat protocol for cervical intraepithelial neoplasia in a hospital population. METHODS: A prospective study was carried out on 285 women with using cytology and colposcopy followed by large loop excision of the transformation zone when indicated as a one step procedure. RESULTS: There was a concordance between colposcopic and cytologic diagnosis. Eighteen women underwent large loop excision. Over treatment rate was 61 per cent but 80 per cent of them occurred in the low grade squamous intraepithelial lesions. The over treatment rate in high grade lesions was 20 per cent. INTERPRETATION & CONCLUSION: A single step see and treat protocol is a feasible management strategy for high grade squamous intraepithelial lesions. However, patient education is essential before undertaking the procedure.

Cervical intraepithelial neoplasia and human papilloma virus infection in renal transplant recipients.

An association between human papilloma virus (HPV) infection and cervical intraepithelial neoplasia has been well established Renal transplant recipients on long term immunosuppression are prone to viral infection. It is possible that there may be an increased prevalence of papilloma virus infection and associated cervical intraepithelial neoplasia in these women. Prospective study of 42 renal transplant recipients and 41 age and parity matched controls was undertaken to determine whether HPV infection and cervical intraepithelial neoplasia (CIN) occurred more often in renal allograft recipients and to assess the relative risk. All women underwent Pap smear, colposcopy, histological examination of biopsy specimen and polymerase chain reaction for HPV 16. Cytology did not pick up HPV infection in any of the women. Colposcopy revealed HPV infection in 15 and CIN in 14 women in the immunosuppressed group and HPV in 7 and CIN in 5 women in control group. Histological evidence of HPV was found in 24 and CIN in 10 women in immunosuppressed group and HPV infection in 13 and CIN in 3 women in control group, giving an odds ratio of 6.1. More women in the immunosuppressed group had CIN of higher degree as well. PCR revealed infection by HPV 16 in 17 cases and 14 controls giving an odds ratio of 1.3. Therefore renal allograft recipients on immunosuppression should be screened by colposcopy and directed biopsy at regular intervals.

Management of prolonged pregnancy: a randomized trial of induction of labour and antepartum foetal monitoring.

BACKGROUND: The two methods of management of prolonged pregnancy, induction of labour and expectant management with foetal surveillance, have pros and cons. Therefore, we compared the induction of labour with serial antenatal foetal monitoring in the management of post-term pregnancy. METHODS: Seventy-four women with uncomplicated pregnancy at 41 weeks (287 days) of gestation were randomly assigned to undergo either induction of labour or serial antenatal foetal monitoring. Labour was induced in the latter group whenever there was evidence of foetal compromise. Antenatal monitoring consisted of the foetal kick count, non-stress test and biophysical profile. RESULTS: Fifty-seven per cent of women went into spontaneous labour by 41 weeks and 4 days (291 days) of gestation and only 14% developed foetal compromise before that. However, when the gestational age was more than 41 weeks and 4 days (291 days), the incidence of meconium staining of amniotic fluid and evidence of uteroplacental insufficiency increased significantly. The rate of caesarean section, instrumental delivery, foetal distress and duration of labour did not differ significantly between the two groups. CONCLUSION: The policy of inducing labour at 41 weeks and 4 days (291 days of gestation) in uncomplicated pregnancies is justified in our population. However, foetal monitoring should begin at 41 weeks of gestation.

CCK-B/Gastrin receptor transmembrane domain mutations selectively alter synthetic agonist efficacy without affecting the activity of endogenous peptides.

Recent efforts have focused on identifying small nonpeptide molecules that can mimic the activity of endogenous peptide hormones. Understanding the molecular basis of ligand-induced receptor activation by these divergent classes of ligands should expedite the process of drug development. Using the cholecystokinin-B/gastrin receptor (CCK-BR) as a model system, we have recently shown that both affinity and efficacy of nonpeptide ligands are markedly affected by amino acid alterations within a putative transmembrane domain (TMD) ligand pocket. In this report, we examine whether residues projecting into the TMD pocket determine the pharmacologic properties of structurally diverse CCK-BR ligands, including peptides and synthetic peptide-derived partial agonists (peptoids). Nineteen mutant human CCK-BRs, each including a single TMD amino acid substitution, were transiently expressed in COS-7 cells and characterized. Binding affinities as well as ligand-induced inositol phosphate production at the mutant CCK-BRs were assessed for peptides (CCK-8 and CCK-4) and for peptoids (PD-135,158 and PD-136, 450). Distinct as well as overlapping determinants of peptide and peptoid binding affinity were identified, supporting that both classes of ligands, at least in part, interact with the CCK-BR TMD ligand pocket. Eight point mutations resulted in marked increases or decreases in the functional activity of the synthetic peptoid ligands. In contrast, the functional activity of both peptides, CCK-8 and CCK-4, was not affected by any of the CCK-BR mutations. These findings suggest that the mechanisms underlying activation of G-protein-coupled receptors by endogenous peptide hormones versus synthetic ligands may markedly differ.

Hyperinsulinemia in polycystic ovary disease.

OBJECTIVE: To evaluate the prevalence of hyperinsulinemia and insulin resistance in women with polycystic ovary disease (PCOD). STUDY DESIGN: Forty women with clinical and biochemical evidence of PCOD and 20 with regular menstrual cycles were studied prospectively. All women underwent a three-hour oral glucose tolerance test following a 100-g glucose load. Plasma sugar and insulin levels were measured. RESULTS: The one-, two- and three-hour insulin values were significantly higher in women with PCOD. The sum insulin, cumulative insulin, peak insulin and area under the insulin response curve were similarly higher in women with PCOD than in the controls. The presence of hirsutism was more often associated with hyperinsulinemia and insulin resistance, but body mass index and menstrual irregularity were not. CONCLUSION: Hyperinsulinemia and insulin resistance seem to be commonly associated with PCOD.

Maternal zinc indices and small babies.

BACKGROUND: Maternal zinc deficiency has been reported to be associated with foetal growth retardation. This study aimed to determine if zinc deficiency is associated with foetal growth retardation in south Indian women. METHODS: A prospective study was undertaken to evaluate the maternal zinc indices in those bearing small-for-gestational age babies and in those with appropriate-for-gestational age babies. Zinc levels in plasma, red blood cells and white blood cells in both groups were assayed in 65 patients with small-for-gestational age babies (regardless of cause) and 51 women with appropriate-for-gestational age babies. RESULTS: There was no significant difference in the mean (SD) plasma [67.5 (9) v. 70.67 (13.9)], red blood cell [47.26 (5.8) v. 45.69 (8.2)] and white blood cell [55.61 (10.5) v. 54.77 (12.4)] zinc levels in mothers who gave birth to small-for-gestational age babies and those who delivered appropriate-for-gestational age babies. The presence of predisposing factors for intrauterine growth retardation also did not influence the maternal zinc levels. CONCLUSION: Maternal zinc levels were not associated with intrauterine growth retardation in our population.

The Griess test: an inexpensive screening test for asymptomatic bacteriuria in pregnancy.

This study was designed to compare urine microscopy and the Griess test as screening tests for asymptomatic bacteriuria (AB) in pregnancy, using urine culture as a gold standard. Urine microscopy, Griess test and urine culture were carried out on urine samples from 483 antenatal women. AB was detected by urine culture in 30 (6.8%) of the patients. The Griess test and urine microscopy were found to have a sensitivity of 63.3% and 60%, specificity of 99.5% and 83.6%; and positive predictive value of 90.5% and 21.1% respectively. Most of the infections missed by the Griess test were either <100,000 colony count on culture or caused by gram positive cocci. The cost of the Griess test was only one tenth of that of urine microscopy. The Griess test is a simple and inexpensive test that when compared to urine microscopy has equal sensitivity, and better specificity and positive predictive value for the detection of AB in pregnancy than urine microscopy.

Detection of CIN by naked eye visualization after application of acetic acid.

A prospective study was undertaken to determine the sensitivity and specificity of acetic application to the cervix followed by naked eye visualization as a screening test for detection of cervical intraepithelial neoplasia. Three hundred and seventy two sexually active woman in the reproductive age group were studied. All the women underwent Papanicolaou test, acetic acid test and colposcopy. One hundred and seventy five woman were acetic acid test negative, 197 women were acetic acid test positive. The sensitivity of acetic acid test was 72.4%, specificity 54% and false negative rate 15.2%, as compared to papanicolaou test which had a sensitivity of 13.2%, specificity of 96.3% and false negative rate of 24.4%. The advantage of the acetic acid test lies in its easy technique, low cost and high sensitivity which are important factors for determining the efficacy of any screening programme in developing countries.

Hypertension in pregnancy. Outcomes, proteinuric vs. nonproteinuric.

OBJECTIVE: To study perinatal and maternal outcome in pregnancies with proteinuric and nonproteinuric hypertension and, on the basis of our observations, to formulate guidelines for management of women with non-proteinuric hypertension. STUDY DESIGN: Review of charts of 216 women with hypertensive disorders of pregnancy over a period of 18 months was carried out. Statistical analysis was performed using the chi 2 test. RESULTS: Nonproteinuric hypertension generally occurred after 37 weeks of gestation, and in 64% of cases the diastolic blood pressure remained < 100 mm Hg. Only 15% required antihypertensives, and perinatal mortality and small-for-dates infants were significantly less common when compared to those in the proteinuric group. CONCLUSION: Proteinuria should be considered an important marker of perinatal outcome. Women with nonproteinuric hypertension may be managed in the outpatient clinic. Monitoring by serial ultrasound scan and cardiotocography is required only in selected cases.

Sclerosing stromal tumour of the ovary. A case report.

Sclerosing stromal tumors (SST) of the ovary are a distinct, but rare benign neoplasms. These tumors appear solid and are very vascular and therefore give the impression of a malignant tumor. They occur mostly in young women. Morphologically and histologically they have distinct characteristics, which make them different from other stromal tumors. The importance of this report is that, though the tumor appears malignant (solid and vascular) since it occurs in young women, care should be taken before embarking on radical surgery.