Maternal and perinatal outcome of pregnancy in women with one previous caesarean section-a study at a tertiary care centre

Background: Increasing rates of primary caesarean section has led to an increased proportion of obstetric population with history of prior caesarean delivery. There is growing concern by obstetrician for optimizing the management of these high risk cases. The present study was undertaken to evaluate obstetric and fetal outcome of patients presenting at term with history of one previous LSCS.Methods: This was a prospective hospital based observational study conducted at Vani Vilas Hospital and Bowring and Lady Curzon Hospital, Department of OBG, BMC and RI, Bangalore. The study included 300 patients who had undergone previous one LSCS with term pregnancy.Results: Majority of patients, that is 186 (62%) were in the age group of 21 to 25 years. Out of 300 patients, 94 (31.33%) patients went for repeat LSCS without trial. 206 (68.67%) patients were included in the trial of labour group, out of which 109 (52.9%) patients had successful vaginal delivery. 97 (47.1%) patients went for repeat LSCS in trial group due to various indications, commonest being scar tenderness.Conclusions: Delivery of patients with previous caesarean section should always be conducted in a well-equipped hospital where facilities for immediate intervention are available if necessity arises. These patients should be counselled antenatally regarding institutional delivery, encouraging trial of labour after caesarean section in select group of patients with close fetal and maternal monitoring for early detection of complications and its management reduces maternal and perinatal mortality and morbidity

Cancer of the uterine cervix and human papillomavirus infection

Human papillomaviruses (HPVs) have emerged as the principal sexually transmitted causal agents in the development of cancer of the uterine cervix in women. They also cause a variety of benign lesions, warts, intraepithelial neoplasia and anogenital, oral and pharyngeal papillomas. Presently, more than 100 HPV genotypes have been identified in humans, and about one-third of them have been sequenced. Of these, while HPV types 16 and 18 are considered to be the high-risk types, HPV 6 and 11 are the low-risk types in the development of cervical cancer. Evidence for causal role of HPV in the development of cervical neoplasia comes from the etiological and epidemiological observations together with the experimental findings of the molecular pathways elicited by HPV-transforming genes. Further evidence in favour of papillomavirus as the carcinoma virus comes from the findings of presence of HPV infections in cancers of oral, esophageal, larynx and nonmelanoma skin cancers. The oncogenic potentials of the virus have been attributed to its E6 and E7 genes. The products of these two genes stimulate cell proliferation by activating the cell-cycle-specific proteins and interfere with the functions of cellular growth-regulatory proteins, p53 and Rb. Identification and characterization of several human pathogenic HPV types warrant prevention of viral infection through vaccination or therapeutic intervention which could eventually control infection and expression of human pathogenic papillomaviruses

A simple 'paper smear' method for dry collection, transport and storage of cervical cytological specimens for rapid screening of HPV infection by PCR

Human papillomaviruses (HPVs) are major pathogens associated with the development of cancer of the uterine cervix, the most common malignant tumour of women world-wide. Reliable diagnosis of HPV infection, particularly the 'high-risk' types (16/18), may facilitate early identification of 'high-risk' populations for developing cervical cancer and may augment the sensitivity and specificity of primary cervical cancer screening programmes by complementing the conventional Pap test. A simple paper smear method has been developed for dry collection, transport and storage of cervical smears/scrapes at room temperature for subsequent detection of HPV DNA by PCR assay. Imprint biopsies, blood and fine-needle aspirates were also collected by this method. The cervical scrapes or other body fluids were smeared (within 0.5-1 cm diameter) and dried on to sterile small slides made of Whatman 3MM filter paper, and stored individually at room temperature or at 4°C. A small piece (2-3 mm) of the paper smear was punched or cut out with a sterile surgical blade, boiled in an eppendorf tube containing 50 µl of distilled water for 5 min and used directly for PCR amplification. The quality and quantity of DNA derived from paper smears and the results of PCR amplifications for HPV type 16, BRCA1 and p53 genes were identical to those obtained from the same samples following collection in PBS, storage (-70°C) and phenol-chloroform-based DNA extraction. DNA was stable in the paper smears for up to a year, whether stored at room temperature or at 4°C. This method is simple, rapid and cost-effective, and can be effectively employed for large-scale population screening, especially for regions where the specimens are to be transported from distant places to the laboratory

Detection of human papillomavirus DNA sequences in cancer of the urinary bladder by in situ hybridisation and polymerase chain reaction

Objective: To evaluate the prevalence of "high risk" human papillomavirus type 16 (HPV 16) in transitional cell carcinoma of the urinary bladder. Materials and Methods: The study included 10 biopsy specimens from male patients of transitional cell carcinoma of the urinary bladder for the detection of HPV DNA sequences. Specimens were collected from the Urology Clinic of the K.G. Medical College Hospital, Lucknow, India. Detection of HPV DNA was carried out by tissue in situ hybridisation (a single copy gene localisation method) using 3H-labelled HPV DNA probe and also by polymerase chain reaction (PCR) techniques using primers to HPV 16 upstream regulatory region (URR). RESULTS--Out of 10 cases of transitional cell carcinoma of the urinary bladder, "high risk" HPV 16 DNA was detected only in one (10%) by using in situ hybridisation whereas two cases (20%) were found to be positive by polymerase chain reaction. Conclusion: Our results suggest that the rare occurrence of HPV in bladder carcinoma may not have a causal relation with the viral infection

Human papillomavirus DNA sequences in adenocarcinoma of the uterine cervix in Indian women

Background: Infection with human papillomavirus (HPV) is considered to be the principal causal agent in the development of squamous cell carcinoma of the uterine cervix. Although adenocarcinoma of the cervix originates adjacent to the squamous epithelial neoplastic lesions, the etiopathogenesis of adenocarcinoma is not yet clearly understood. Recent studies have raised more controversy, rather than answering the question of whether specific HPV infection also plays a role in the development of adenocarcinoma of the cervix. Molecular DNA hybridization techniques were used to detect HPV types prevalent in both adenocarcinoma and squamous cell carcinoma of the uterine cervix, which is the most common cancer in Indian women. Methods: Histologically confirmed, formaldehydefixed, paraffin-embedded tissue sections from 12 cases of adenocarcinoma and 30 cases of squamous cell carcinoma of the uterine cervix were analyzed retrospectively or the presence of HPV DNA types 6b, 11, 16, and 18 by Southern blot hybridization and in situ hybridization. Results: Of 12 adenocarcinomas, 5 (41.67%) tumors were positive for HPV DNA. All five cases were positive for HPV 16, and two (16.6%) of these were hybridized again to the HPV 18-specific DNA probe. All tumors were negative for HPV 6b and 11. In addition, no biopsy specialnens were positive after hybridization with a mixed probe of HPV 31, 33, 35, 39, and 45. These results were compared to those obtained for 30 squamous cell carcinomas of the cervix. Although 20 (66%) were exclusively positive for HPV 16 and 6 (20%), more tumors were of HPV 16 related types as detected under nonstringent conditions of hybridization, only one (3%) was positive for HPV 18. The results of in situ hybridization were found to be in good agreement with those of Southern blotting. Conclusions: HPV 16 is the type present almost exclusively in squamous cell carcinoma of Indian women. A higher frequency of HPV 16 in adenocarcinoma of Indian women, in contrast to HPV 18, as reported from other regions, may be attributed to geographic variation rather than to histologic differences only, and both HPV 16 and 18 may be present in adenocarcinoma of the uterine cervix

Fibroset™ and neuromuscular pain: a multicentric, real world, observational, post-marketing surveillance study in Indian patients suffering from neuromuscular pain

Background: Neuromuscular disease (NMD) is a condition due to abnormality or damage to muscles and nerves causing painful symptoms. Symptomatic management involves use of conventional painkillers, but desirable relief is not achieved due to multimodal pathophysiology of disease. This study evaluated the efficacy and safety of Fibroset™ tablets in subjects with neuromuscular pain. Methods: Subjects with neuromuscular pain, previously unsatisfied with standard therapies, were enrolled. Subjects were advised to take Fibroset™ one tablet BID for 2 weeks with their standard therapy. Efficacy was evaluated on pain, stiffness, swelling, weakness, tenderness, and difficulty in activity of daily living (ADL) as per the visit schedule. Tolerability of therapy was also evaluated. Results: 59 patients were enrolled in study and 46 patients were included in the final analysis. Fibroset™ supplementation significantly reduced all evaluated parameters (p<0.05 vs baseline). The mean pain score from 2.50 to 0.89, while mean stiffness score was reduced to 0.55 from 1.87 at end of study. The mean swelling score was reduced to 0.81 from 2.04, while the mean weakness score was reduced to 0.64 from baseline score of 1.79. The mean tenderness score was reduced from baseline score of 1.90 to 0.65 and the mean ADL score was reduced to 0.63 from baseline score of 2.00. No treatment related side effects were observed. Conclusions: Fibroset™ is a potentially effective and safe therapy for subjects with neuromuscular pain. It can be used to reduce symptoms in patients with unsatisfactory results with conventionally standard care therapy

Intracellular growth of Mycobacterium tuberculosis after macrophage cell death leads to serial killing of host cells

A hallmark of pulmonary tuberculosis is the formation of macrophage-rich granulomas. These may restrict Mycobacterium tuberculosis (Mtb) growth, or progress to central necrosis and cavitation, facilitating pathogen growth. To determine factors leading to Mtb proliferation and host cell death, we used live cell imaging to track Mtb infection outcomes in individual primary human macrophages. Internalization of Mtb aggregates caused macrophage death, and phagocytosis of large aggregates was more cytotoxic than multiple small aggregates containing similar numbers of bacilli. Macrophage death did not result in clearance of Mtb. Rather, it led to accelerated intracellular Mtb growth regardless of prior activation or macrophage type. In contrast, bacillary replication was controlled in live phagocytes. Mtb grew as a clump in dead cells, and macrophages which internalized dead infected cells were very likely to die themselves, leading to a cell death cascade. This demonstrates how pathogen virulence can be achieved through numbers and aggregation states. DOI: http://dx.doi.org/10.7554/eLife.22028.00

Touchdown General Primer (GP5+/GP6+) PCR and optimized sample DNA concentration support the sensitive detection of human papillomavirus

BACKGROUND: The GP5+/GP6+ PCR assay is a well-established HPV detection technique. This study has examined the effects of incorporating 'hot start' and 'touchdown' steps into the protocol. In addition, dTTP was substituted with dUTP to permit contamination control measures against carry-over PCR product. METHODS: Firstly, HPV-16 was amplified from SiHa cell DNA (0.1 ng–100 ng) diluted in a background of C-33A DNA (100 ng-2 μg). Secondly, the detection of small quantities (15ag-1.5pg) of HPV recombinant plasmids (types 16, 31, 33, 45, 51, 52, and 56) diluted in C-33A DNA was investigated. Thirdly, clinical sample DNA extracts (cervical smears, formalin-fixed vaginal lesions and breast tumors) were tested for HPV. Six different PCR protocols were assessed. HPV was detected by gel electrophoresis, and by Southern and dot blot hybridization. RESULTS: HPV detection sensitivity was dependent on the total amount of DNA in a PCR. Touchdown protocols supported HPV-16 detection from 1 ng or 0.5 ng SiHa cell DNA in a background of 2 μg or 1 μg C-33A DNA respectively, and from 0.1 ng of SiHa cell DNA (~28 copies HPV-16) in 500 ng or 100 ng background DNA. Under standard GP5+/GP6+ annealing conditions, HPV-16 went undetected when the DNA content of a PCR was 2 μg or 1 μg, and with 500 ng C-33A DNA the sensitivity limit was 1 ng SiHa cell DNA. HPV recombinant plasmids were each detected with high (albeit varying) sensitivity by a touchdown protocol. HPV-31 was better amplified under standard annealing conditions (1.5fg in 100 ng background DNA) than by a touchdown approach (15fg detection limit). HPV-52 was not amplified by the standard protocol at the dilutions tested. Seventeen different HPV types were demonstrated in 47/65 (72%) abnormal cytology samples recorded as HPV negative by standard GP5+/GP6+ conditions. Twenty-one different HPV types were recorded in 111/114 (97%) vaginal lesions. Multiple infections were also detectable using a touchdown approach. Of 26 breast tumors, 5 (19%) tested HPV positive by the standard assay and 15/26 (58%) using a touchdown protocol. CONCLUSION: Touchdown modification of the GP5+/GP6+ PCR assay enables the detection of HPV undetected under regular assay conditions. The use of standardized DNA quantities in a PCR rather than standard sample volumes containing arbitrary amounts of DNA is supported. A touchdown approach may be beneficial as an analytical test for the re-evaluation of (apparently) HPV negative abnormal cervical cytological or histological samples, and for investigating the association of HPV with disease conditions at diverse organ sites. The clinical utility of a touchdown approach for HPV detection requires further investigation as increased assay analytical sensitivity may not necessarily equate with improved clinical sensitivity or specificity