Pre-induction cervical assessment using transvaginal ultrasound versus Bishops cervical scoring as predictors of successful induction of labour in term pregnancies: A hospital-based comparative clinical trial.

OBJECTIVE: To evaluate the association between transvaginal ultrasound scan of cervix and Bishop's score in predicting successful induction of labour, cut-off points and patients' tolerability and acceptance for both procedures. DESIGN: A comparative clinical trial. SETTING: A tertiary hospital in Selangor, Malaysia. PARTICIPANTS: 294 women planned for elective induction of labour for various indications were included. All women had transvaginal ultrasound to assess the cervical length and digital vaginal examination to assess the Bishop cervical scoring by separate investigators before induction of labour. PRIMARY OUTCOME MEASURE: To evaluate the association of the cervical length by transvaginal ultrasound scan and Bishop score in predicting successful induction of labour. SECONDARY OUTCOME MEASURE: Variables associated with successful induction of labour and patients' tolerability and acceptance for transvaginal ultrasound scan of cervix. RESULTS: There was no statistically significant difference among the vaginal and Caesarean delivery groups in terms of mean maternal age, height, weight, body mass index, ethnicity and gestational age at induction. Vaginal delivery occurred in 207 women (70.4%) and 87 women (29.6%) delivered via Caesarean section. There was a high degree of correlation between the cervical length and Bishop score (r-value 0.745; p <0.001). Sonographic assessment of cervical length demonstrated a comparable accuracy in comparison to Bishop score. Analysis using ROC curves noted an optimal cut-off value of ≤27mm for cervical length and Bishop score of ≥ 4, with a sensitivity of 69.1% vs 67%, specificity 60.9% vs 55%, and area under the curves (AUCs) of 0.672 and 0.643 respectively (p <0.001). Multivariate logistic regression analysis demonstrated that parity (OR 2.70), cervical length (OR 0.925), Bishop score (OR 1.272) and presence of funnelling (OR 3.292) were highly significant as independent predictors of success labour induction. Women also expressed significantly less discomfort with transvaginal ultrasound compared with digital vaginal examination. CONCLUSION: Sonographic assessment of cervical measurement predicts the success of induction of labour with similar diagnostic accuracy with conventional Bishop score.

Rapid Multiplex Loop-Mediated Isothermal Amplification (m-LAMP) Assay for Differential Diagnosis of Leprosy and Post-Kala-Azar Dermal Leishmaniasis.

Leprosy and post-kala-azar dermal leishmaniasis (PKDL) are co-endemic neglected tropical diseases often misdiagnosed because of close resemblance in their clinical manifestations. The test that aids in differential diagnosis of leprosy and PKDL would be useful in endemic areas. Here, we report development of a multiplex loop-mediated isothermal amplification (m-LAMP) assay for differential detection of Mycobacterium leprae and Leishmania donovani using a real-time fluorometer. The m-LAMP assay was rapid with a mean amplification time of 15 minutes, and analytical sensitivity of 1 fg for L. donovani and 100 fg for M. leprae. The distinct mean Tm values for M. leprae and L. donovani allowed differentiation of the two organisms in the m-LAMP assay. Diagnostic sensitivity of the assay was evaluated by using confirmed cases of leprosy (n = 40) and PKDL (n = 40) (tissue and slit aspirate samples). All the leprosy and PKDL samples used in this study were positive by organism-specific QPCR and loop-mediated isothermal amplification assays. The diagnostic sensitivity of the m-LAMP assay was 100% (95% CI: 91.2-100.0%) for detecting PKDL and 95% for leprosy (95% CI: 83.1-99.4%). Our m-LAMP assay was successfully used to detect both M. leprae and L. donovani in a patient coinfected with leprosy and macular PKDL. The m-LAMP assay is rapid, accurate, and applicable for differential diagnosis of leprosy versus PKDL, especially in endemic areas.

Development of a rapid loop-mediated isothermal amplification assay for diagnosis and assessment of cure of Leishmania infection.

BACKGROUND: Leishmaniasis is a spectrum of diseases with great relevance to public health. Conventional diagnostic methods are time consuming, needing trained personnel. A robust, rapid and cost effective diagnostic test is warranted for on-time diagnosis and field application. METHODS: We have developed a loop mediated isothermal amplification (LAMP) assay with primers (n = 6) based on Leishmania donovani kDNA for detection of Leishmania infection, using a closed tube to prevent cross-contamination. The assay was used to detect Leishmania infection in biological samples obtained from patients of visceral leishmaniasis (VL), post kala-azar dermal leishmaniasis (PKDL) and cutaneous leishmaniasis (CL). RESULTS: The assay was positive for L. donovani, L. tropica and L. major parasites, with the highest sensitivity towards L. donovani (1 fg DNA). The high sensitivity of the assay for detection of L. donovani was reflected in its ability to detect parasite DNA within 30 min of amplification time with a threshold detection limit of ≥25 copies per reaction. The assay detected parasite in 64 of 66 VL blood samples (sensitivity, 96.9%; 95% CI: 89.6-99.2%), 15 of 15 VL bone marrow aspirate samples (sensitivity, 100%; 95% CI:79.6-100%), 65 of 67 PKDL tissue biopsy samples (sensitivity, 97%; 95% CI:89.7-99.2%). The assay was evaluated in a few cases of CL wherein it was found positive in 8 of 10 tissue biopsies (sensitivity, 80%; 95% CI: 49-94.3%). The assay was negative in all control blood (n = 76) and tissue biopsy (n = 24) samples (specificity, 100%; 95% CI: 96.3-100%). Further, the assay was evaluated for its utility in assessment of cure in treated VL and PKDL patients. The assay detected parasite DNA in 2 of 20VL blood samples and 2 of 21 PKDL tissue samples. Out of 4 cases that were positive for parasite DNA at post treatment stage, 2 patients (1VL and 1 PKDL) returned with relapse. CONCLUSIONS: The study demonstrated a Leishmania genus specific closed tube LAMP assay for reliable and rapid molecular diagnosis of VL and PKDL with potential for application in assessment of cure.

Percutaneous retrogasserian glycerol rhizotomy for trigeminal neuralgia: A simple, safe, cost-effective procedure.

OBJECTIVE: Trigeminal neuralgia (TN) is a condition that has been studied over decades and whose pathogenesis has still not been well defined. Various open and minimally invasive procedures are in vogue for the treatment of intractable TN. All these procedures have their complications and recurrence rates. Percutaneous retrogasserian glycerol rhizotomy (PRGR) is one of the minimally invasive procedures that have been popular for quite a long time. MATERIAL AND METHODS: This paper is a prospective study analyzing the results of 93 patients with refractory TN who were treated with PRGR. RESULTS: There was an immediate pain relief in 96.8% of patients and long-term pain relief in 89.4% of patients, with a mean follow-up duration of 18.8 months. Recurrence of pain was seen in 10.4% of patients. CONCLUSIONS: The PRGR is a simple, safe, cost-effective procedure without any need for expensive equipment and with a good outcome that is compared to the other relatively more expensive open and minimally invasive procedures.

Assessing intake of spices by pattern of spice use, frequency of consumption and portion size of spices consumed from routinely prepared dishes in southern India.

BACKGROUND: Measurement of dietary intake of spices is gaining significance because of recognition of their health promoting benefits as well as its use for risk assessment of contaminant exposures. Estimating intake of spices at the individual level, presents several challenges since various spices are used as an integrated part of a prepared food and consumed in amounts much smaller than other dietary components. The objective of the present study is to assess intake of spices at the household and individual level on the basis of pattern of spice use and portion size of spice consumed from routinely prepared dishes in Hyderabad city in Southern India. METHODS: The study was conducted in 100 households in urban areas of Hyderabad city in India with the help of a spice intake questionnaire that was prepared to collect information on the pattern of spice use, frequency, and quantity of spice consumption of 17 spices routinely used in Indian cuisine. The quantity of spice intake was assessed by measuring portion size of spice consumed from the quantity of i) spices added in routinely prepared dishes and ii) the prepared dish consumed by an individual. RESULTS: Based on the type of dish prepared and frequency of preparing the dishes, 11 out of 17 spices were found to be consumed by more than 50% of the households. Maximum number of spices was consumed at weekly frequencies. Red chillies and turmeric were the most frequently consumed spices by 100% of the households. The mean total intake of spices was observed to be higher through dishes consumed daily (10.4 g/portion) than from those consumed at weekly or monthly frequencies. Highest portion size intake was observed for chillies (mean 3.0 g; range 0.05-20.2 g) and lowest for nutmeg (mean 0.14 g; range 0.02-0.64 g) and mace (mean 0.21 g; range: 0.02-0.6 g). CONCLUSIONS: The study suggested that assessment of intake of spices varies with frequency of use of spices and type of dish consumed. Portion size estimations of spices consumed and the frequency of consumption of the spice containing dishes facilitates in quantifying spice intake at the individual level.

Assessment of reactivity of three treponemal tests in non-treponemal non-reactive cases from sexually transmitted diseases clinic, antenatal clinic, integrated counselling and testing centre, other different outdoor patient departments/indoor patients of a tertiary care centre and peripheral health clinic attendees.

In India, many state reference centres for sexually transmitted infections perform only a single screening assay for syphilis diagnosis. In this study, Treponema pallidum haemagglutination (TPHA) was performed on 1115 Venereal Disease Research Laboratory (VDRL)/rapid plasma regain (RPR) non-reactive and 107 reactive sera out of 10,489 tested by VDRL/RPR according to the National AIDS Control Organisation syphilis testing protocol. A total of 47 Specimens reactive in TPHA and non-reactive with VDRL test were subjected to fluorescent treponemal antibody absorption and enzyme-immunoassay. Seroprevalence considering both VDRL and TPHA positivity was highest (4.4%) in sexually transmitted diseases clinic attendees than in other subject groups. Positivity by two treponemal tests in 24 (2.2%) cases non-reactive by VDRL/RPR was representative of the fully treated patients or latent or late syphilis cases. The findings highlight that a suitable treponemal confirmatory test should be performed in all the diagnostic laboratories.

Validation of a simple resazurin-based promastigote assay for the routine monitoring of miltefosine susceptibility in clinical isolates of Leishmania donovani.

Simple, cost-effective approach for routine surveillance of parasite susceptibility to antileishmanial drug miltefosine (MIL) is highly desirable for controlling emergence of drug resistance in visceral leishmaniasis (VL). We validated a simple resazurin-based fluorimetric assay using promastigotes to track natural MIL tolerance in Leishmania donovani parasites from VL cases (n = 17) against standard amastigote assay, in two different labs in India. The inter-stage MIL susceptibility correlated strongly (r = 0.70, p = 0.0018) using J774.A.1 macrophage cell line-based amastigote assay and fluorescence-based resazurin assay for promastigotes. Investigation of inter-stage MIL susceptibility for the same set of clinical isolates in another lab also showed a strong correlation (r = 0.72, p = 0.0012) using mouse peritoneal macrophages for amastigote assay and resazurin-based alamar blue assay for promastigotes. Additionally, parasites from post-kala-azar dermal leishmaniasis (PKDL) lesions (n = 7, r = 0.78, p = 0.046) and MIL-induced parasites (r = 0.92, p = 0.0001; n = 3) also exhibited a strongly correlated inter-stage miltefosine susceptibility. Thus, our results support the utility of resazurin assay as a simplified biological tool for MIL susceptibility monitoring in clinical isolates from MIL-treated VL/PKDL patients.

Comparative assessment of CDS, CLSI disc diffusion and Etest techniques for antimicrobial susceptibility testing of Neisseria gonorrhoeae: a 6-year study.

BACKGROUND: A variety of techniques are available for antimicrobial susceptibility testing of Neisseria gonorrhoeae. OBJECTIVE: The aim of this study was to find a cost-effective, reliable and easily applicable microbiological method to detect antimicrobial susceptibilities of N. gonorrhoeae in resource-poor countries. DESIGN: Prospective study. SETTING: Male and female STD clinic of Regional STD Teaching, Training and Research Centre, New Delhi, India. PARTICIPANTS: N. gonorrhoeae isolates from all male and female patients presenting with acute gonococcal urethritis and cervical discharge. MATERIAL AND METHODS: A total of 295 consecutive N. gonorrhoeae isolates during 2005-2010 was used to compare the Clinical and Laboratory Standards Institute (CLSI) and CDS disc diffusion technique with Etest by performing antimicrobial susceptibility testing in parallel for penicillin, tetracycline, ceftriaxone, ciprofloxacin and spectinomycin. WHO reference strains were used as controls. RESULTS: CDS disc diffusion zones of inhibition showed that complete percentage agreement for penicillin, ciprofloxacin and tetracycline was high with their analogous Etest minimal inhibitory concentrations in comparison to CLSI disc diffusion technique, that is, 91.5%, 92.9% and 99.3% versus 87.5%, 88.5% and 74.9%, respectively. CDS results had less number of major and minor category discrepancies in comparison to CLSI and CDS method showed excellent correlation coefficient (r=1) with Etest for all five antimicrobial agents tested in comparison to CLSI (r=0.92). It was very poor (r=0.61) by CLSI method for tetracycline. The correlation coefficients between the two methods and the Etest were identical if tetracycline was removed from the CLSI analysis. CONCLUSIONS: The CDS technique is an attractive alternative for N. gonorrhoeae susceptibility testing and is recommended for monitoring the antimicrobial susceptibility in less developed and resource-poor settings to facilitate enhanced antimicrobial resistance surveillance when the WHO Gonococcal Antimicrobial Surveillance Programme is undergoing expansion to meet the ongoing challenges of surveillance and control of gonococcal antimicrobial resistance.

Potential role of FDG PET imaging in predicting metastatic potential and assessment of therapeutic response to neoadjuvant chemotherapy in Ewing sarcoma family of tumors.

AIMS AND OBJECTIVES: The aim of this study was to retrospectively correlate FDG uptake in primary Ewing sarcoma family of tumors (ESFT) with tumor behavior, and to evaluate whether FDG PET can be used to predict response to neoadjuvant chemotherapy (NACT) in this patient group. METHODS: Out of the total 54 patients of recently diagnosed ESFT who underwent pretreatment FDG PET imaging, group I included patients without metastasis at presentation (n = 34) and group II included those with metastasis at presentation (n = 20). Fourteen of these patients had undergone FDG PET after 4 cycles of induction chemotherapy and surgical resection of primary tumor. In this subgroup of 14 patients, maximum standardized uptake value (SUVmax) of primary tumor was estimated before and after 4 cycles of induction chemotherapy and was correlated with the histopathological response in terms of necrosis in the tumor specimen. RESULTS: Mean SUVmax in the primary tumor in group I patients was 6.84 and in group II patients, it was 11.31. The difference between mean SUVmax of these 2 groups was significant by Wilcoxon test analysis, with P < 0.01. In group II patients, SUVmax in metastasis with maximum FDG uptake was consistently lower as compared with that of primary tumor. In subgroup of 14 patients, Pearson correlation analysis showed that percentage change in SUVmax of primary tumor correlated well with percentage necrosis on histopathological examination (P < 0.01). CONCLUSION: FDG uptake in primary ESFT reflected its metastatic potential and hence the aggressive behavior. The significant correlation between change in metabolic activity of the primary tumor and histopathological response after neoadjuvant chemotherapy suggests that FDG PET may be an ideal noninvasive method to assess tumor behavior and response to therapy in ESFT.

In-hospital observation after antibiotic switch in pneumonia: a national evaluation.

PURPOSE: To evaluate the clinical benefit of in-hospital observation after the switch from intravenous (IV) to oral antibiotics in a large Medicare population. Retrospective studies of relatively small size indicate that the practice of in-hospital observation after the switch from IV to oral antibiotics for patients hospitalized with community-acquired pneumonia (CAP) is unnecessary. METHODS: We performed a retrospective examination of the US Medicare National Pneumonia Project database. Eligible patients were discharged with an ICD-9-CM diagnosis consistent with community-acquired pneumonia and divided into 2 groups: 1) a "not observed" cohort, in which patients were discharged on the same day as the switch from IV to oral antibiotics and 2) an "observed for 1 day" cohort, in which patients remained hospitalized for 1 day after the switch from IV to oral antibiotics. We compared clinical outcomes between these 2 cohorts. RESULTS: A total of 39,242 cases were sampled, representing 4341 hospitals in all 50 states and the District of Columbia. There were 5248 elderly patients who fulfilled eligibility criteria involving a length of stay of no more than 7 hospital days (2536 "not observed" and 2712 "observed for 1 day" patients). Mean length of stay was 3.8 days for the "not observed" cohort and 4.5 days for the "observed for 1 day" cohort (P <.0001). There was no significant difference in 14-day hospital readmission rate (7.8% in the "not observed" cohort vs 7.2% "observed for 1 day" cohort, odds ratio 0.91; 95% confidence interval [CI] 0.74-1.12; P =.367) and 30-day mortality rate (5.1% "not observed" cohort vs 4.4% in the "observed for 1 day" cohort, odds ratio 0.86; 95% CI, 0.67-1.11; P =.258) between the "not observed" and "observed for 1 day" cohorts. CONCLUSIONS: Our analysis of the US Medicare Pneumonia Project database provides further evidence that the routine practice of in-hospital observation after the switch from IV to oral antibiotics for patients with CAP may be avoided in patients who are clinically stable although these findings should be verified in a large randomized controlled trial.

Is exercise stress testing a cost-saving strategy for risk assessment of pediatric wolff-Parkinson-white syndrome patients?

BACKGROUND: In Wolff-Parkinson-White syndrome (WPW) patients the loss of pre-excitation in a single heartbeat during exercise stress testing (EST) is a predictor of low risk of sudden death. The purpose of this study was to: 1) assess the frequency of loss of pre-excitation in a single heartbeat during exercise testing, and 2) compare the cost of EST versus trans-catheter electrophysiology study (EPS) in the risk assessment of WPW patients. METHODS: A retrospective review of 50 cases of patients with WPW who underwent EST was conducted including demographics, history of supraventricular tachycardia, associated congenital heart disease, maximum heart rate achieved, and loss of pre-excitation in a single heartbeat. Hospital costs of EST and EPS were compared. RESULTS: Of the 50 patients who underwent EST, 4 (8%), lost pre-excitation in a single heartbeat during EST. No differences were found regarding gender, age at diagnosis or EST, history of supraventricular tachycardia, presence of congenital heart disease or maximal heart rate. A cost comparison, utilizing the cost data: EST ($62.75) and EPS ($5,597) found EST to be a cost-saving approach in WPW patients. With 4 patients losing pre-excitation during EST, the cost saving of EST was $22,388 for this population of WPW patients. CONCLUSIONS: A frequency of 8% loss of pre-excitation was found in a pediatric sample that underwent EST. Additionally, EST was shown to be a cost-saving strategy in risk assessment of pediatric WPW patients.

Development of a large-scale chemogenomics database to improve drug candidate selection and to understand mechanisms of chemical toxicity and action.

Successful drug discovery requires accurate decision making in order to advance the best candidates from initial lead identification to final approval. Chemogenomics, the use of genomic tools in pharmacology and toxicology, offers a promising enhancement to traditional methods of target identification/validation, lead identification, efficacy evaluation, and toxicity assessment. To realize the value of chemogenomics information, a contextual database is needed to relate the physiological outcomes induced by diverse compounds to the gene expression patterns measured in the same animals. Massively parallel gene expression characterization coupled with traditional assessments of drug candidates provides additional, important mechanistic information, and therefore a means to increase the accuracy of critical decisions. A large-scale chemogenomics database developed from in vivo treated rats provides the context and supporting data to enhance and accelerate accurate interpretation of mechanisms of toxicity and pharmacology of chemicals and drugs. To date, approximately 600 different compounds, including more than 400 FDA approved drugs, 60 drugs approved in Europe and Japan, 25 withdrawn drugs, and 100 toxicants, have been profiled in up to 7 different tissues of rats (representing over 3200 different drug-dose-time-tissue combinations). Accomplishing this task required evaluating and improving a number of in vivo and microarray protocols, including over 80 rigorous quality control steps. The utility of pairing clinical pathology assessments with gene expression data is illustrated using three anti-neoplastic drugs: carmustine, methotrexate, and thioguanine, which had similar effects on the blood compartment, but diverse effects on hepatotoxicity. We will demonstrate that gene expression events monitored in the liver can be used to predict pathological events occurring in that tissue as well as in hematopoietic tissues.

Superiority of denaturing high performance liquid chromatography over single-stranded conformation and conformation-sensitive gel electrophoresis for mutation detection in TSC2.

We evaluated denaturing high pressure liquid chromatography (DHPLC) as a scanning method for mutation detection in TSC2, and compared it to conformation-sensitive gel electrophoresis (CSGE) and single-stranded conformation polymorphism analysis (SSCP). The first 20 exons of TSC2 were amplified from 84 TSC patients and screened initially by CSGE and then by DHPLC. Optimization of DHPLC analysis of each exon was carried out by design of primers with minimum variation in the melting temperature of the amplicon, and titration of both elution gradient and temperature. CSGE analysis identified 40 shifts (21 unique) in the 84 patients and 20 exons. All of these variants were detected by DHPLC, and an additional 27 changes (14 unique) were identified. Overall 15 of 28 (54%) unique single base substitutions were detected by CSGE; all were detected by DHPLC. 25 definite or probable mutations were found in these 84 patients (30%) in exons 1-20 of TSC2. In a subsequent blinded analysis of 15 samples with 18 distinct TSC2 sequence variants originally detected by SSCP in another centre, all variants were detected by DHPLC except one where the variation occurred within the primer. Ten other (7 unique) sequence variants were detected in these samples which had not been detected by SSCP. Overall, 11 of 16 (69%) unique single base substitutions were detected by SSCP; all were detected by DHPLC. We conclude that DHPLC is superior to both CSGE and SSCP for detection of DNA sequence variation in TSC2, particularly for single base substitution mutations.

Practical problems in the management of leprosy.

The categorization of leprosy into paucibacillary or multibacillary depends on the report of slit-skin smears. Unfortunately, in many control programmes the quality of slit smears is below par. Taking the example of India, the main reasons were that the work of laboratory technicians was unrewarding as compared to serving in a general health care system. There was lack of equipment and an unrealistic patient to technician ratio. Future attempts were made by experienced workers to devise a clinical system for classifying leprosy as paucibacillary or multibacillary based on counting the number of lesions. However this method did not prove cost-effective because more paucibacillary patients were classified in the multibacillary group increasing the burden of treatment. A renewed attempt to improve slit-smear performance should be made by modifying the existing methods. This can definitely improve the situation. Patients with multiple macular lesions and those with neuritic leprosy are best treated with the MB-MDT regimen. The treatment for PB leprosy is to continue up to 6 months but in MB leprosy with a high bacterial index a longer duration of MDT may be required. Following completion of MDT many cases with deformity are accumulating and their care forms are a neglected part of many control programmes. In addition to strengthening the infrastructure, simple techniques must be imparted to those with deformities and disabilities. This involves the artful and innovative cooperation of the health worker, patient and the community. The leprosy worker should be motivated to promote such activities.

The Economic Impact of a Foodborne Disease Outbreak Due to Staphylococcus aureus.

The economic cost of a Staphylococcus aureus outbreak involving over 100 persons was estimated. About 41% of the expenditure was borne by the affected persons which includes, loss of wages or productivity loss and other expenses. The cost of hospitalization, laboratory investigations, etc., was 43%. Educative and preventive measures would considerably reduce the economic cost of the outbreaks which are very high for a developing economy. A comparison of the economic cost calculated on the basis of percent of per capita income with that of a similar outbreak in the United States indicated that the cost of a foodborne disease outbreak is higher in India than in the United States.