Health-seeking pathway of drug-resistant TB patients in Vadodara, India.

BACKGROUND: Health-seeking behaviour refers to patients' choices regarding their preferred healthcare destination and the timing of seeking assistance for treatment. Patients with TB usually first approach the private sector and/or lose several months' time in inappropriate diagnosis and treatment due to lack of awareness regarding the availability of standard treatment protocols. This can lead to poor outcomes such as drug-resistant TB (DR-TB) and/or death. METHODOLOGY: A cross-sectional study was conducted to examine the health-seeking pathway and delays in diagnosis and initiation of DR-TB treatment among patients registered with the DR-TB centre in Vadodara District (India). RESULTS: A total of 93 patients were enrolled in the study; the median age was 35 years (IQR 24-45). For the first visit, 59 (63%) patients chose a public healthcare facility, mainly because the facility was near their residence (n = 20, 21.5%). The median delay in reaching the first healthcare facility was 12 days (IQR 7.5-30). Delay in reaching second- and third-level care was respectively 25 days (IQR 9-68) and 16 days (IQR 4-67). CONCLUSION: Two-thirds of patients required visits to a second healthcare centre for diagnosis, while one third needed a third visit. The overall median delay for reaching the DR-TB centre was 60 days (IQR 26-122). The median duration from symptom onset to the first healthcare contact fell within the timeframe for screening symptoms in standard diagnosis.

CONTEXTE: Le comportement de recherche de santé fait référence aux choix des patients concernant leur destination préférée en matière de soins de santé et le moment où ils recherchent de l'aide pour le traitement. Les patients atteints de TB s'adressent généralement en premier lieu au secteur privé et/ou perdent plusieurs mois en diagnostics et traitements inappropriés en raison d'un manque d'information sur la disponibilité des protocoles de traitement standard. Cela peut conduire à des résultats médiocres tels que la TB résistante aux médicaments (DR-TB, pour l'anglais « drug-resistant TB ¼) et/ou la mort. MÉTHODOLOGIE: Une étude transversale a été menée pour examiner le parcours de recherche de soins et les retards dans le diagnostic et l'initiation du traitement de la DR-TB parmi les patients enregistrés au centre de DR-TB dans le district de Vadodara (Inde). RÉSULTATS: Au total, 93 patients ont participé à l'étude ; l'âge médian était de 35 ans (IQR 24­45). Pour la première visite, 59 patients (63%) ont choisi un établissement de santé public, principalement parce que l'établissement était proche de leur résidence (n = 20 ; 21,5%). Le délai médian pour atteindre le premier établissement de santé était de 12 jours (IQR 7,5­30). Le délai pour atteindre le deuxième et le troisième niveau de soins était respectivement de 25 jours (IQR 9­68) et de 16 jours (IQR 4­67). CONCLUSION: Deux tiers des patients ont dû se rendre dans un deuxième centre de soins pour obtenir un diagnostic, tandis qu'un tiers a dû se rendre dans un troisième centre. Le délai médian global pour atteindre le centre de lutte contre la TB était de 60 jours (IQR 26­122). La durée médiane entre l'apparition des symptômes et le premier contact avec les soins de santé correspondait au délai de dépistage des symptômes dans le cadre du diagnostic standard.

HostSeq: a Canadian whole genome sequencing and clinical data resource.

HostSeq was launched in April 2020 as a national initiative to integrate whole genome sequencing data from 10,000 Canadians infected with SARS-CoV-2 with clinical information related to their disease experience. The mandate of HostSeq is to support the Canadian and international research communities in their efforts to understand the risk factors for disease and associated health outcomes and support the development of interventions such as vaccines and therapeutics. HostSeq is a collaboration among 13 independent epidemiological studies of SARS-CoV-2 across five provinces in Canada. Aggregated data collected by HostSeq are made available to the public through two data portals: a phenotype portal showing summaries of major variables and their distributions, and a variant search portal enabling queries in a genomic region. Individual-level data is available to the global research community for health research through a Data Access Agreement and Data Access Compliance Office approval. Here we provide an overview of the collective project design along with summary level information for HostSeq. We highlight several statistical considerations for researchers using the HostSeq platform regarding data aggregation, sampling mechanism, covariate adjustment, and X chromosome analysis. In addition to serving as a rich data source, the diversity of study designs, sample sizes, and research objectives among the participating studies provides unique opportunities for the research community.

Microplastics in environment: global concern, challenges, and controlling measures.

Plastic pollution in various forms has emerged as the most severe environmental threat. Small plastic chunks, such as microplastics and nanoplastics derived from primary and secondary sources, are a major concern worldwide due to their adverse effects on the environment and public health. Several years have been spent developing robust spectroscopic techniques that should be considered top-notch; however, researchers are still trying to find efficient and straightforward methods for the analysis of microplastics but have yet to develop a viable solution. Because of the small size of these degraded plastics, they have been found in various species, from human brains to blood and digestive systems. Several pollution-controlling methods have been tested in recent years, and these methods are prominent and need to be developed. Bacterial degradation, sunlight-driven photocatalyst, fuels, and biodegradable plastics could be game-changers in future research on plastic pollution control. However, recent fledgling steps in controlling methods appear insufficient due to widespread contamination. As a result, proper regulation of environmental microplastics is a significant challenge, and the most equitable way to manage plastic pollution. Therefore, this paper discusses the current state of microplastics, some novel and well-known identification techniques, strategies for overcoming microplastic effects, and needed solutions to mitigate this planetary pollution. This review article, we believe, will fill a void in the field of plastic identification and pollution mitigation research.

Delay and attrition before treatment initiation among MDR-TB patients in five districts of Gujarat, India.

Setting: Gujarat, a state in west India. Background: Although treatment initiation has been improving among patients diagnosed with multidrug-resistant tuberculosis (MDR-TB) in programme settings, it has still not reached 100%. Objectives: To determine pre-treatment attrition (not initiated on treatment within 6 months of diagnosis), delay in treatment initiation (>7 days from diagnosis) and associated factors among MDR-TB patients diagnosed in 2014 in five selected districts served by two genotypic drug susceptibility testing (DST) facilities and a drug-resistant TB centre in Gujarat. Design: This was a retrospective cohort study involving record review. Results: Among 257 MDR-TB patients, pre-treatment attrition was seen in 20 (8%, 95%CI 5-12). Patients with 'follow-up sputum-positive' as their DST criterion and sputum smear microscopy status 'unknown' at the time of referral for DST were less likely to be initiated on treatment. The median delay to treatment initiation was 8 days (interquartile range 6-13). Patients referred for DST from medical colleges were more likely to face delays in treatment initiation. Conclusion: The Gujarat TB programme is performing well in initiating laboratory-confirmed MDR-TB patients on treatment. However, there is further scope for reducing delay.

Contexte : Dans le Gujarat, un état de l'ouest de l'Inde, même si la mise en route du traitement a été améliorée pour les patients ayant eu un diagnostic de tuberculose multirésistante (TB-MDR) dans le contexte des programmes, elle n'a pas encore atteint 100%.Objectif : Déterminer l'attrition avant traitement (c'est-à-dire un traitement pas mis en route dans les 6 mois suivant le diagnostic), le retard à la mise en route (>7 jours du diagnostic) et les facteurs associés parmi des patients TB-MDR diagnostiqués en 2014 dans cinq districts sélectionnés servis par deux structures de test génotypique de pharmacosensibilité (DST) et un centre de TB résistante au Gujarat.Schéma : Ceci a été une étude rétrospective de cohorte basée sur une revue de dossiers.Résultats : Sur 257 patients TB-MDR, l'attrition avant traitement a été constatée chez 20 patients (8% ; IC95% 5­12). Les patients ayant un « crachat de suivi positif ¼ comme critère de DST et un statut de microscopie de frottis de crachats « inconnu ¼ lors de la référence pour DST ont été moins susceptibles d'être mis sous traitement. Le délai médian de mise en route du traitement a été de 8 jours (intervalle interquartile 6­13). Les patients référés pour DST de centres hospitalières universitaires sont plus susceptibles de rencontrer des retards à la mise en route du traitement.Conclusion : Le programme TB du Gujarat est performant en mettant en route le traitement de TB-MDR confirmé par le laboratoire. Il reste cependant une marge d'amélioration en matière de réduction des délais.

Marco de referencia: Guyarat es un estado del occidente de la India donde se han logrado avances en la iniciación del tratamiento de los pacientes con diagnóstico de tuberculosis multirresistente (TB-MDR) en el marco programático, pero aún no se ha alcanzado el 100%.Objetivos: Determinar la tasa de abandono anterior al tratamiento (no haber iniciado tratamiento en un lapso de 6 meses después del diagnóstico), el retraso en la iniciación del tratamiento (>7 días después del diagnóstico) y los factores asociados en los pacientes diagnosticados con TB-MDR de cinco distritos escogidos de Guyarat atendidos por dos centros de pruebas genotípicas de sensibilidad a los medicamentos (DST) y un centro de tuberculosis resistente en el 2014.Método: Fue este un estudio de cohortes retrospectivo con examen de las historias clínicas.Resultados: De los 257 pacientes con diagnóstico de TB-MDR, se observó un abandono anterior al tratamiento en 20 casos (8%; IC95% 5­12). La probabilidad de iniciar el tratamiento era menor en los pacientes cuyo criterio para practicar las DST era 'seguimiento a la positividad del esputo' y su situación de la baciloscopia del esputo era desconocida en el momento de la remisión para las pruebas. La mediana del retraso en la iniciación del tratamiento fue 8 días (amplitud intercuartil 6­13). Los pacientes remitidos de las facultades de medicina para realizar las DST presentaban con mayor frecuencia retrasos en la iniciación del tratamiento.Conclusión: El desempeño del programa contra la TB de Guyarat es adecuado con respecto a la iniciación del tratamiento de los pacientes con TB-MDR confirmada por el laboratorio. Sin embargo, aún son necesarios progresos en esta esfera con el fin de acortar los retrasos.

Why are HIV-infected people not started on antiretroviral therapy? A mixed-methods study from Gujarat, India.

Setting: Five purposively selected antiretroviral therapy (ART) centres in Gujarat, India. Objectives: To assess the proportion of ART-eligible people living with the human immunodeficiency virus (PLHIV) who were not initiated on ART within 2 months of being recorded as eligible, to identify factors associated with non-initiation and to explore reasons from the provider's perspective. Design: We used a mixed-methods design (triangulation) of 1) a quantitative phase involving record reviews and cohort analysis (Poisson regression) of PLHIV registered during April 2014-March 2015, and 2) a qualitative phase involving one-to-one interviews with 25 providers. Results: Of 2079 ART-eligible PLHIV, 339 (16%) were not started on ART within 2 months. PLHIV with CD4 counts of <350 cells/µl and patients who were labourers, hospitalised, bedridden or registered with certain ART centres were more likely not to be initiated on ART. Qualitative results were categorised into two broad themes: government health system- and patient-related challenges, which validated and complemented the quantitative findings. Conclusion: Several patient subgroups at greater risk of ART non-initiation were identified, along with reasons for risk; this has important programme implications for achieving the UNAIDS 90-90-90 goal, and particularly the second 90 component of having 90% of diagnosed PLHIV start ART.

Contexte : Cinq centres du TAR (traitement antirétroviral) sélectionnés dans ce but dans l'état de Gujarat, Inde.Objectifs : Evaluer la proportion de personnes vivant avec le virus de l'immunodéficience humaine (PVVIH) éligibles pour le TAR non mis sous TAR dans les 2 mois de leur éligibilité, identifier les facteurs associés à la non initiation et explorer les raisons vues par les prestataires de soins.Schéma: Nous avons eu recours à un mélange de méthodes (triangulation) : 1) une phase quantitative impliquant une revue des dossiers et une analyse de la cohorte (régression de Poisson) des PVVIH enregistrés entre avril 2014 et mars 2015, et 2) une phase qualitative impliquant des entretiens individuels avec 25 prestataires de soins.Résultats : Sur 2079 PVVIH éligibles au TAR, 339 (16%) n'ont pas été mis sous traitement dans les 2 mois. Les PVVIH ayant un taux de CD4 <350 cellules/µl, les patients qui étaient des travailleurs journaliers, hospitalisés, alités ou suivis par certains centres du TAR ont été plus susceptibles de ne pas être mis sous TAR. Les résultats qualitatifs ont été classés en deux vastes catégories : système de santé du gouvernement et défis liés aux patients ; ceux-ci ont validé et complété les résultats quantitatifs.Conclusion : Plusieurs sous-groupes de patients ayant un risque plus élevé de non mise en route du TAR et les raisons de ce problème ont été identifiés ; ceci pourrait avoir des implications importantes pour le programme dans l'atteinte de l'objectif 90­90­90, surtout en ce qui concerne le deuxième 90, qui consiste à débuter le TAR chez 90% des PVVIH diagnostiqués.

Marco de referencia: Cinco centros de suministro del tratamiento antirretrovírico (TAR) de Gujarat en la India, escogidos por muestreo dirigido.Objetivos: Evaluar la proporción de personas positivas frente al virus de la inmunodeficiencia humana (PPVIH) aptas para recibir el TAR, que no habían iniciado el tratamiento 2 meses después de haberse considerado idóneas; determinar los factores asociados con la falta de iniciación del TAR; y analizar las razones desde la perspectiva de los profesionales de salud.Método: Se utilizó un diseño de métodos mixtos (triangulación), con una fase cuantitativa de análisis de las historias clínicas y de cohortes de PPVIH registradas de abril del 2014 a marzo del 2015 y una fase cualitativa con entrevistas personales a 25 profesionales de salud.Resultados: De las 2079 PPVIH aptas para recibir el TAR, 339 no lo habían iniciado en un lapso de 2 meses (16%). La probabilidad de no iniciar el TAR fue mayor en las PPVIH con cifras de linfocitos CD4 <350 células/µl, los pacientes que eran obreros, estaban hospitalizados, encamados o que acudían a determinados centros de suministro de TAR. Los resultados se clasificaron en dos amplias categorías, a saber: problemas relacionados con el sistema público de salud o atribuibles a los pacientes, con lo cual se validaron y complementaron los resultados cuantitativos.Conclusión: Varios subgrupos de pacientes presentaron un mayor riesgo de no iniciar el TAR y se determinaron las razones del riesgo; los resultados pueden tener repercusiones importantes en el programa y favorecer el progreso hacia el cumplimiento del triple objetivo 90­90­90, sobre todo de su segundo componente, según el cual el 90% de las PPVIH debe iniciar el TAR.

The impact of age on clinical outcomes following cardiac resynchronisation therapy.

BACKGROUND: Cardiac resynchronisation therapy (CRT) is an established treatment for selected patients with symptomatic left ventricular (LV) systolic dysfunction. Heart failure (HF) is primarily a disease of the elderly; however, these patients are underrepresented in CRT trials. Our aim was to evaluate the impact of age on clinical outcomes following CRT. METHODS: A consecutive series of 177 patients was identified and divided into those aged ≤ 7 5 years (n = 131, mean ± SD 62.1 ± 11.2 years) and those aged >75 years (n = 46, mean ± SD 80.7 ± 4.1 years). The primary end point was a composite of all-cause mortality or HF hospitalisation. RESULTS: During a median ± IQR follow up of 28.5 ± 33.7 months, the event rate for the primary end point was significantly higher in the elderly compared to younger patients (20.1 vs. 11.1 %, respectively, logrank p = 0.020). This was mainly driven by an excess mortality rate among those aged >75 years (10 vs. 4.7%, respectively, logrank p = 0.018) whereas HF hospitalisation rates were similar between groups (10 vs. 6.4%, respectively, logrank p = 0.301). After adjusting for comorbidities and ICD status, the difference in the composite end point rates was attenuated and no longer significant (HR 1.580, 95% CI 0.899-2.778; p = 0.112 for >75 vs. ≤ 75 years). Notably, both groups demonstrated similar response rates to CRT in terms of symptomatic improvement, reverse LV remodelling and neurohormonal activation. CONCLUSIONS: CRT is equally effective in the elderly as in younger patients to reduce adverse clinical outcomes. For those who fulfil the prerequisite selection criteria, it should be considered as a valid therapeutic option.

Tobacco sales and marketing within 100 yards of schools in Ahmedabad City, India.

OBJECTIVES: The Government of India passed the Cigarettes and Other Tobacco Products (Prohibition of Advertisement and Regulation of Trade and Commerce, Production, Supply and Distribution) Act, 2003 (COPTA 2003), which prohibits the sale of tobacco products within 100 yards of educational institutions and regulates tobacco advertising. The aim of this research was to monitor compliance with the section of COPTA 2003 regarding the advertisement, display and sale of tobacco products around educational institutions in Ahmedabad City, India. STUDY DESIGN: Observational study around 30 randomly selected schools. METHODS: In March 2010, an observational study was conducted to assess compliance with COPTA 2003 in Ahmedabad City, India. All vendors within a 100-yard radius of 30 randomly selected schools were identified. At locations where tobacco was sold, information was collected regarding type of product sold, sale of tobacco in single units and advertising. RESULTS: Twenty public schools and 10 private schools were sampled. Of these, 87% [n = 26, 95% confidence interval (CI) 69-95%] had tobacco sales within 100 yards of their entrance. Of the 771 vendors observed, 24% (n = 185, 95% CI 18-32%) sold tobacco products. Tobacco advertising in violation of the law was found around 57% of schools (n = 17, 95% CI 39-73%), product displays around 83% of schools (n = 25, 95% CI 65-93%) and single sales around 70% of schools (n = 21, 95% CI 51-84%). CONCLUSIONS: Violation of the sections of COPTA 2003 regarding sale of tobacco products around educational institutions and advertising in general is widespread in Ahmedabad City, India. Effective enforcement of the existing law is necessary to protect the children in India from widespread exposure to the sale and marketing of tobacco products.

A qualitative and quantitative assessment of the impact of three processing algorithms with halving of study count statistics in myocardial perfusion imaging: filtered backprojection, maximal likelihood expectation maximisation and ordered subset expectation maximisation with resolution recovery.

INTRODUCTION: Ordered subset expectation maximisation with depth-dependent resolution recovery (OSEM-RR) is a processing algorithm reported to improve images with halved tracer activity in myocardial perfusion scintigraphy (MPS) compared to filtered backprojection (FBP) using conventional activities. OSEM-RR has not yet been compared with maximal likelihood expectation maximisation (MLEM). METHODS: 39 patients undergoing MPS and two anthropomorphic phantoms (one with, one without an inferior wall insert) had full-time (FT) and half-time (HT) SPECT datasets acquired simultaneously and processed by FBP, MLEM and OSEM-RR. Two experienced reporters scored images of all clinical studies (n=234) for conspicuity of a perfusion defect, with results being compared using Wilcoxon paired and Kappa tests. A quantitative assessment based on mean segmental pixel counts taken from numbers automatically displayed over the 20 segments of Cedars Sinai Autoquant QPS image were compared using Pearson's correlation and Bland Altman analysis. RESULTS: A small but consistent superior concurrence between FT and HT datasets for OSEM-RR compared to FBP and MLEM was observed for both qualitative and quantitative analyses. OSEM-RR resulted in better definition of the inferior wall defect on the phantom study. CONCLUSION: OSEM-RR appears superior to both FBP and MLEM in terms of handling reduced count statistics.

Impact of the RNTCPIRL-EQA-OSE visits on quality of sputum smear microscopy services of Gujarat, India.

OBJECTIVE: On-site evaluation of laboratories with standard checklist is a first step to promote effective and consistent supervision. The present study was carried out to evaluate the impact of the RNTCP- Intermediate Reference Laboratory External Quality Assessment- On-Site Evaluation visits on quality of sputum smear microscopy services of Gujarat, India. Data of three IRL-EQA-OSE visit rounds, carried out between January 2005 and December 2010 are presented here. MATERIAL AND METHODS: Within the Revised National Tuberculosis Control Programme EQA framework, the IRL, Ahmedabad visited all Gujarat District Tuberculosis Centres, and evaluated their sputum smear microscopy services. The study covered a cohort of 29 DTCs during each of the three IRL-EQA-OSE visits. The authors focused on section III of Annexure A to study and analyse the said impact. In order to convert qualitative data into quantitative one, the authors denoted a score of 1 to "Acceptable" (No Error) remark and 0 to "Not-Acceptable" (Error) one. RESULTS: A larger degree of improvement was noted in Standard Operating Procedure practices, Disinfection practices, and Internal Quality Control practices. Many DTCs did not retrain their laboratory staff in EQA methodology. The Gujarat DTCs achieved an overall score of (820/957) 86% during the initial OSE visits which consistently improved to (842/957) 88% and (885/957) 92% during the two follow-up OSE visits along with sustenance and improvement in many important laboratory parameters. CONCLUSION: The co-sponsoring organisation (IRL) recognises the challenges and therefore, is committed to supporting state-level implementation of EQA through additional training, technical assistance to districts, and improving this technical guidance. By periodic IRL-EQA-OSE visits, sputum smear microscopy services can be sustained and improved at field level.

Ascorbate prevents placental oxidative stress and enhances birth weight in hypoxic pregnancy in rats.

This study isolated the effects of maternal hypoxia independent of changes in maternal nutrition on maternal circulatory and placental molecular indices of oxidative stress and determined whether maternal antioxidant treatment conferred protection. Pregnant rats were subjected to normoxic pregnancy or 13% O2 chronic hypoxia for most of gestation with and without maternal treatment with vitamin C in the drinking water. Maternal hypoxia with and without vitamin C did not affect maternal food or water intake and led to a significant increase in maternal and fetal haematocrit. At gestational day 20, maternal plasma urate and L-cysteine concentrations, and placental levels of 4-hydroxynonenal and heat shock protein 70 were increased while placental heat shock protein 90 levels were decreased in hypoxic pregnancy. The induction of maternal circulatory and placental molecular indices of oxidative stress in hypoxic pregnancies was prevented by maternal treatment with vitamin C. Maternal hypoxia during pregnancy with or without vitamin C increased placental weight, but not total or compartmental volumes. Maternal treatment with vitamin C increased birth weight in both hypoxic and normoxic pregnancies. The data show that maternal hypoxia independent of maternal undernutrition promotes maternal and placental indices of oxidative stress, effects that can be prevented by maternal treatment with vitamin C in hypoxic pregnancy. While vitamin C may not be the ideal candidate of choice for therapy in pregnant women, and taking into consideration differences in ascorbic acid metabolism between rats and humans, the data do underlie that antioxidant treatment may provide a useful intervention to improve placental function and protect fetal growth in pregnancy complicated by fetal hypoxia.

Differential response of cancer cells to HDAC inhibitors trichostatin A and depsipeptide.

BACKGROUND: Over the last decade, several drugs that inhibit class I and/or class II histone deacetylases (HDACs) have been identified, including trichostatin A, the cyclic depsipeptide FR901228 and the antibiotic apicidin. These compounds have had immediate application in cancer research because of their ability to reactivate aberrantly silenced tumour suppressor genes and/or block tumour cell growth. Although a number of HDAC inhibitors are being evaluated in preclinical cancer models and in clinical trials, little is known about the differences in their specific mechanism of action and about the unique determinants of cancer cell sensitivity to each of these inhibitors. METHODS: Using a combination of cell viability assays, HDAC enzyme activity measurements, western blots for histone modifications, microarray gene expression analysis and qRT-PCR, we have characterised differences in trichostatin A vs depsipeptide-induced phenotypes in lung cancer, breast cancer and skin cancer cells and in normal cells and have then expanded these studies to other HDAC inhibitors. RESULTS: Cell viability profiles across panels of lung cancer, breast cancer and melanoma cell lines showed distinct sensitivities to the pan-inhibitor TSA compared with the class 1 selective inhibitor depsipeptide. In several instances, the cell lines most sensitive to one inhibitor were most resistant to the other inhibitor, demonstrating these drugs act on at least some non-overlapping cellular targets. These differences were not explained by the HDAC selectivity of these inhibitors alone since apicidin, which is a class 1 selective compound similar to depsipeptide, also showed a unique drug sensitivity profile of its own. TSA had greater specificity for cancer vs normal cells compared with other HDAC inhibitors. In addition, at concentrations that blocked cancer cell viability, TSA effectively inhibited purified recombinant HDACs 1, 2 and 5 and moderately inhibited HDAC8, while depsipeptide did not inhibit the activity of purified HDACs in vitro but did in cellular extracts, suggesting a potentially indirect action of this drug. Although both depsipeptide and TSA increased levels of histone acetylation in cancer cells, only depsipeptide decreased global levels of transcriptionally repressive histone methylation marks. Analysis of gene expression profiles of an isogenic cell line pair that showed discrepant sensitivity to depsipeptide, suggested that resistance to this inhibitor may be mediated by increased expression of multidrug resistance genes triggered by exposure to chemotherapy as was confirmed by verapamil studies. CONCLUSION: Although generally thought to have similar activities, the HDAC modulators trichostatin A and depsipeptide demonstrated distinct phenotypes in the inhibition of cancer cell viability and of HDAC activity, in their selectivity for cancer vs normal cells, and in their effects on histone modifications. These differences in mode of action may bear on the future therapeutic and research application of these inhibitors.

Proficiency panel testing--a reliable tool in external quality assessment of sputum smear microscopy services in Gujarat, India.

OBJECTIVE: To assess the proficiency of Senior TB Laboratory Supervisors (STLSs) and district level Laboratory Technicians (LTs) in sputum smear microscopy. METHOD: Intermediate Reference Laboratory (IRL), Ahmedabad had manufactured and validated Proficiency Panel Testing slides from sputum samples, made On Site Evaluation (OSE) visits of District TB Centres (DTCs) in two rounds, and conducted Proficiency Panel Testing of STLSs & DTC-LTs from January 2005 to June 2009. RESULTS: High level of concordance in Z-N smear grading was found between Microbiologist and district laboratory staff. DTC readers reported overall consistency level of more than 98% in Z-N grade agreement during both the IRL, EQA, OSE visits. The tendency to over-grade the panel slides was much higher (more than 22%) as compared to under-grade (less than 2%) them in "correct slides". High False Positive (HFP) error was not observed in the present study. CONCLUSION: Laboratory supervisor's proficiency can be quickly assessed by Proficiency Panel Testing, under multi-level quality assurance network system of sputum smear microscopy in public health programmes like the RNTCP. Proficiency Panel Testing is highly replicable and reproducible tool for quick and reliable assessment of proficiency of the staff and it can be made more effective by raising the proportion of lower grade positive slides in panel set of each reader. DTC readers' overall agreement level of more than 98% in Z-N grade suggests high level of precision and excellent consistency during both the IRL, EQA, OSE rounds. It is concluded that even for a large network of sputum smear microscopy centres under public health programmes like the RNTCP in order to take corrective action, Proficiency Panel Testing can be effectively used for quick identification of suboptimal- technical performance of the supervisory staff.

Punch biopsy: a useful adjunct in a rapid diagnosis breast clinic.

INTRODUCTION: Triple assessment of breast lesions usually involves the use of core biopsy (CB) or fine needle aspiration cytology (FNAC). Punch Biopsy (PB) is a technique widely used by dermatologists and can be used in superficial breast lesions with dermal involvement. We studied the utilization of PB in a rapid diagnosis breast clinic. METHOD: We reviewed patients who underwent a PB over a seven and a half-year period from December 2001 to May 2009. The indications for biopsy and the contribution of PB to final diagnosis were studied. RESULTS: The commonest indications were breast lump with skin involvement or ulceration (n = 27), suspected Paget's disease (n = 25), discolouration of breast skin (n = 23), and nodules in the breast skin or surgical scar (n = 18). Final diagnosis was benign in 80 patients and malignant in 20. In 74 patients with benign and 7 patients with malignant diagnoses, PB was the only source of histological diagnosis. CONCLUSION: PB is a valuable adjunct to conventional methods of tissue diagnosis such as CB and FNAC in both benign and malignant breast lesions.

Lateralization of cortical function in swallowing: a functional MR imaging study.

BACKGROUND AND PURPOSE: While functional MR imaging and other techniques have contributed to our knowledge of functional brain localization, these methods have not been extensively applied to the complex and incompletely understood task of swallowing. We used functional MR imaging to investigate motor cortex activity during swallowing in healthy human adults. METHODS: Eight subjects were imaged on a 1.5-T MR system using blood oxygen level-dependent contrast mechanisms. Subjects performed three swallowing tasks and a finger-tapping task. Areas of activation in the cortex and subcortical areas were tabulated, and a laterality index, defined as LI = [sigmas left - sigmas right]/[sigmas left + sigmas right] x 100, was computed for the three tasks. RESULTS: Activation was observed in the primary motor and sensory cortices, motor processing and association areas, and subcortical sites. This activity was dominant for one hemisphere with left hemispheric dominance more prevalent among the subjects. Right hemispheric dominance, however, showed stronger lateralization than the left hemisphere. CONCLUSION: Our data indicate that specific sites in the motor cortex and other cortical and subcortical areas are activated with swallowing tasks and that hemispheric dominance is a feature of swallowing under these conditions. In addition, we demonstrate the utility of functional MR imaging in the study of the cortical representation of swallowing and suggest a role for functional MR imaging in the diagnosis of dysphagia of cerebral origin.

Chemical transformations in individual ultrasmall biomimetic containers.

Individual phospholipid vesicles, 1 to 5 micrometers in diameter, containing a single reagent or a complete reaction system, were immobilized with an infrared laser optical trap or by adhesion to modified borosilicate glass surfaces. Chemical transformations were initiated either by electroporation or by electrofusion, in each case through application of a short (10-microsecond), intense (20 to 50 kilovolts per centimeter) electric pulse delivered across ultramicroelectrodes. Product formation was monitored by far-field laser fluorescence microscopy. The ultrasmall characteristic of this reaction volume led to rapid diffusional mixing that permits the study of fast chemical kinetics. This technique is also well suited for the study of reaction dynamics of biological molecules within lipid-enclosed nanoenvironments that mimic cell membranes.

Rapid preparation of giant unilamellar vesicles.

We report here a rapid evaporation method that produces in high yield giant unilamellar vesicles up to 50 microns in diameter. The vesicles are obtained after only 2 min and can be prepared from different phospholipids, including L-alpha-phosphatidylcholine (lecithin), dipalmitoleoyl L-alpha-phosphatidylcholine, and beta-arachidonoyl gamma-palmitoyl L-alpha-phosphatidylcholine. Vesicles can be produced in distilled water and in Hepes, phosphate, and borate buffers in the pH range of 7.0 to 11.5 with ionic strengths up to 50 mM. The short preparation time allows encapsulation of labile molecular targets or enzymes with high catalytic activities. Cell-sized proteoliposomes have been prepared in which gamma-glutamyltransferase (EC 2.3.2.2) was functionally incorporated into the membrane wall.

Cell-to-cell scanning in capillary electrophoresis.

A widespread limitation in using cell-based biosensors for repetitive chemical analysis is loss of agonist-induced response caused by receptor desensitization. We overcome this problem by scanning an array of immobilized cells underneath a capillary electrophoresis column outlet. In this way, electrophoretically fractionated components that exit the separation capillary are always directed onto cells previously unexposed to receptor agonists. To demonstrate this concept of response recovery using a scanning format, we have chosen the bradykinin B2 receptor system in the NG108-15 cell line, which is known to undergo desensitization. Whereas four subsequent injections of 250 microM bradykinin separated by 120 s are found to reduce the NG108-15 cell response markedly, scanning to new cells can fully restore the response during the separation. Furthermore, by pretesting individual NG108-15 cells for an agonist response and then later scanning back to the same cell, we achieved a 100% success rate in detecting bradykinin in subsequent electrophoretic separations.

Some histochemical observations on invasive cells in a myopathy induced in rats with 5-hydroxytryptamine.

The enzyme cytochemistry and proportion of mononuclear cells exclusively present within degenerating muscle fibres of the soleus of rats 24 h after a single intraperitoneal injection of 5-hydroxytryptamine (5-HT) (10 mg/kg) were studied. The following enzymes displayed strong cytochemical activities within these cells: non-specific esterase (against naphthol AS-D acetate and alpha-naphthylacetate), aminopeptidase (against leucine-4-methoxy-2-naphthylamide) and dipeptidyl peptidase II. A peroxidatic activity was also present, but was found to arise from cytochrome oxidase in phagocytosed muscle mitochondria. The aminopeptidase activity in the monocytic cells increased by roughly 50% between 6 and 24 h after 5-HT administration. Dipeptidyl peptidase II was found, by the use of appropriate inhibitors, to be present in both mononuclear cells and muscle fibres and may be implicated in degrading the basal lamina of muscle fibres, thereby permitting access to the mononuclear cells. The morphological, enzyme histochemical and electron microscopical evidence suggest that the cells which invade muscle fibres in the early stages of a myopathy induced by 5-HT in rats are actively phagocytosing, mature macrophages.