A Rapid Field-Deployable Reverse Transcription-Insulated Isothermal Polymerase Chain Reaction Assay for Sensitive and Specific Detection of Bluetongue Virus.

Bluetongue is a non-contagious, haemorrhagic, Culicoides-borne disease of ruminants. The causative agent, bluetongue virus (BTV), is a member of the Orbivirus genus of the Reoviridae family. So far, 26 BTV serotypes have been identified worldwide. The global distribution of bluetongue has been expanding, and rapid detection of BTV, preferably in the field, is critical for timely implementation of animal movement restrictions and vector control measures. To date, many laboratory-based, molecular assays for detection of BTV have been developed. These methods require the samples to be shipped to a central laboratory with sophisticated instruments and highly skilled technicians to perform the assays, conduct analyses and interpret the results. Here, we report the development and evaluation of a rapid, portable, user-friendly, pan-BTV reverse transcription-insulated isothermal polymerase chain reaction (RT-iiPCR) assay that can potentially be used in low-resource field conditions. The total length of the assay was <60 min, and at the end of the assay, the results were automatically displayed as '+' or '-' without the need for data interpretation. The RT-iiPCR assay detected 36 BTV isolates and two in vitro transcribed RNA samples representing all 26 BTV serotypes. The assay did not cross-react with other animal viruses tested, including two closely related orbiviruses. The analytical sensitivity of the assay was as low as nine copies of in vitro transcribed double-stranded BTV RNA. Analysis of BTV-infected whole blood samples showed that the BTV RT-iiPCR assay was as sensitive as real-time RT-PCR. The assay can potentially be used for rapid screening of animals for BTV in routine diagnostics and for monitoring bluetongue outbreaks both in ruminants and in Culicoides vectors in the field and in the laboratory.

Is Tertiary care Treatment Affordable to All? - Explore Alternative (s) for Healthcare Financing.

BACKGROUND: Health indicators such as life expectancy at birth, infant mortality rate etc. are some of the tools to measure the social development of that country. Due to advancement in technology, disease profile etc. expenditure on health care, especially tertiary care treatment is increasing day by day, and is beyond the reach of common people. This is one of the hindrances in the social development. India is a developing country and approx. 35% population is below poverty line. Since many people now are below poverty line or in a lower income group, people do not have any feeling to save or cannot save the money for future expenditure on health; therefore the tertiary care facility is inaccessible to a major fraction of the society. METHODS: A study was conducted at Sanjay Gandhi Postgraduate Institute of Medical Sciences (SGPGIMS) Lucknow in the month of May-June 2007 with objectives to find out the level of awareness about hospital care financing, analyze the mode and mechanism of Health care financing and to assess the affordability of the tertiary care treatment by the patients undergoing treatment at this centre. A prospective study of 154 patients by floating the structured questionnaire containing 17 questions, covering various issues of HCF was carried out. RESULTS: While observing the HCF pattern of High cost treatment diseases related to ICU, kidney transplant, cardiovascular surgery, Haematology, Surgical Gastroenterology etc. (expenditure limit more than Rupees (Rs) 75000 so far incurred in one patient), the study revealed that the average size of family is 6 members with an income of approx. Rs. 10185 per month per family. Average expenditure so far incurred by one shows that majority 115 (75%) patients bear the expenses themselves (out of pocket) It was also observed that approx. 50% out of pocket expenditure was met by taking loan from the bank, relatives, selling of assets and mortgaging the assets. approx. 34 (22%) reported that their expenditure will be reimbursed or got advance payment. Only 40 patients out of pocket had received some sort of assistance from some of the of the sources like CM, PM Fund and other types of donation. The study further highlights that average treatment expenditure already incurred in one patient i.e. Rupees 219376/- has exceeded the per capita expenditure on health care of individual (Rs. 1500/-), & total expenditure on whole family (Rs. 9000/-) and even total estimated annual income of one member (Rs.20370/) and whole family (approx. 122220/-). Only 2 (1.2%) respondents had health insurance policy. CONCLUSION: The present study findings reflect that the tertiary & high cost treatment is beyond the reach of majority and will have great impact on the economy and health profile of society. In view of above, it is the need of the hour to strengthen the mechanism of the HCF by mass awareness with great emphasis is to be given to meet the "out of pocket expenditure" and encourage the "health insurance" mechanism. This may help in providing the tertiary care treatment to many people; otherwise the treatment cost is unaffordable.

Ambulatory blood pressure monitoring: a useful tool to diagnose hypertension and supervise it's treatment.

Automatic ambulatory blood pressure monitoring (ABPM) for the diagnosis and treatment ofhypertension(HTN) is not common in Nepal. The purpose of this study is to evaluate various characteristics of hypertensive patients undergoing ABPM before starting antihypertensive treatment and evaluate the adequacy of the blood pressure (BP) control during antihypertensive treatment. ABPM was performed in 108 consecutive patients attending the hypertension clinic of Nepal Medical College Teaching Hospital from 1st March 2005 to 30th April 2007 with DynaPulse 5000A (version 3.20q ) for approximately 24 hours. Male female ratio was 59:49 and age (mean +/- SD) was 47.8 +/- 16.4 years. The maximum use of ABPM (25.9%) was noted in the age group of 40-49 years. Body mass index was 25.7 +/- 3.8. Diabetes was noted in 13% patients. Maximum use of ABPM was observed in Newar ethnic group (56.5%). ABPM was used for the diagnosis of HTN in 62.0% patients and for follow up in 38.0% patients. Severe HTN was seen in approximately half (47.2%) of the hypertensive patients. Majority of the patients (88.0%) had dipper type of HTN. Beta-blocker (35.6%), ACE inhibitor/Losartan (31.1%) and calcium channel antagonist (26.7%) were the usual antihypertensive agents used. Single antihypertensive agent was used in the majority of patients (64.1%). In a small number of patients (42, 38.9%) undergoing ABPM during antihypertensive therapy, the adequacy of control of HTN was very poor.

Drug use among the urban poor in Kolkata: behaviour and environment correlates of low HIV infection.

BACKGROUND: HIV infection in injecting drug users (IDUs) has worked as a driving force for further spread of the virus in other population groups. Major metropolitan cities such as Mumbai, Kolkota, Chennai and Delhi have seen a diffusion of injecting drug use within the last decade. The prevalence of HIV infection among injectors ranges from 2% to 30%. Identifying effective interventional elements that have kept the prevalence of HIV low for the past 7 years among IDUs of Kolkata is thus of public health importance. METHODS: A purposive sample of opioid/opiate users was studied. Primary and secondary data on drug users, law-enforcement environment, records at drug treatment centres, jail admission data related to the 'Narcotic Drug and Psychotropic Substance Act' and interventions in other risk groups were collected. Laboratory tests for HIV, hepatitis B surface antigen and syphilis were done on consenting IDUs (n=129) and non-IDUs (n=120). For univariate and multivariate analysis, IDUs were taken as cases and non-IDUs as controls. RESULT: Of the IDUs, 2% were positive for HIV. No non-IDU was HIV-positive. Significantly more non-IDUs (10% v. 4%, p=0.05) were positive for syphilis. Sharing injection equipment within the past 6 months was reported by 71% of IDUs; sharing partners were stable and ranged from 1 to 3. More IDUs compared to non-IDUs reported being in touch with intervention programmes. The police has been tolerant to needle-syringe exchange and oral sublingual buprenorphine substitution conducted in Kolkata. Unlike in the early 1990s, non-IDUs did not switch to injecting during non-availability of brown sugar in the latter half of the 1990s and instead sought tratment. The availability of high quality heroin (>20%-50%) was low and the proportion of moderate quality heroin (>10%-20%) went up during these times due to increased police seizures. No intervention exists in jails despite the fact that a large number of drug users spend time in jail. CONCLUSION: Stable and few injection equipment-sharing partners of IDUs, launching of early targeted interventions among IDUs and sex workers in the city, police tolerance to harm reduction activities and preference of non-IDUs for detoxification during heroin draught periods have kept HIV prevalence at a low level among drug users of Kolkata for the past 7 years. immediate launching of interventions for drug users in jails seems necessary. Similar multi-pronged strategies with targeted and environmental intervention could work in other settings as well.

Interaction of bacteriophage T4 AsiA protein with Escherichia coli sigma70 and its variant.

Bacteriophage T4 produces a small protein AsiA, which inhibits transcription from sigma70-dependent promoters in E. coli by tightly binding to sigma70 and is therefore termed as anti-sigma factor. We observed that there was no inhibition of single round transcription at lac UV5 promoter when AsiA was added to preformed open complex between RNA polymerase and template DNA. However, transcription was found to proceed normally at 'extended -10' promoters in the presence of AsiA. It appears therefore that AsiA binds sigma70 at its 4.2-subdomain or in its close vicinity. Further experiments on immunoprecipitation of sigma70 and a mutant sigma70-V576G with AsiA seem to corroborate such conclusion.

Periplasmic expression of biologically active vesicular stomatitis virus phosphoprotein P in Escherichia coli.

Overexpression of a clone of vesicular stomatitis virus phosphoprotein P (New Jersey serotype) using T7 promoter with phoA leader sequence and a simpler two-step purification procedure of the expressed protein has been developed. The purified protein retains its ability to activate the transcription reaction. Comparative transcriptional assay using the protein P purified from periplasmic space and from cytosol (in the form of inclusion body) of Escherichia coli establishes the fact that the former is 10 times more efficient than the latter in activating the transcription reaction in vitro.

Percutaneous retrogasserian radiofrequency thermal rhizotomy for trigeminal neuralgia.

Forty patients suffering from intractable unilateral trigeminal neuralgia involving more than one division of the trigeminal nerve were treated by percutaneous radiofrequency thermocoagulation of the trigeminal sensory root. The aim of the operation was to relieve the pain without producing dense sensory deficit in the face. This goal was achieved by making selective lesions in the sensory root with gradually increasing temperature 60 degrees C to 90 degrees C. Three to four consecutive lesions each for 60 seconds have been found to produce excellent pain relief in 77.7% with good and fair results in the rest. The recurrence rate has been found to be 15% during 2 years of follow-up. Considerable dysaesthesia was observed in 5% of cases. Corneal anaesthesia was found in 5% cases while one patient developed neuroparalytic keratitis. Transient trigeminal motor weakness was observed in 10% of patients.