Effects of different salinity levels on larval growth and development of disease vectors of Culex species

Aim: To determine species distribution of mosquitoes in coastal brackish wetlands and in paddy field area of Kerala along with the effect of increased salinity on the development and survival of major Japanese encephalitis vectors. Methodology: Mosquito survey was carried out in different sites of coastal brackish water as well as in paddy fields of Alappuzha district. First instar larvae were reared until adult emergence at different constant saline water. Each individual was monitored daily for mortality. Larvae were placed in a plastic container with 200 ml distilled water, which served as controls. Counts of larvae, pupae and adults were recorded every 24 hr up to adult emergence. Results: The collected mosquitoes were Culex tritaeniorhynchus, Cx. gelidus, Cx. sitiens, Cx. quinquefasciatus, Cx. bitaeniorhynchus and Anopheles vagus. During summer season, Cx. sitiens (92.85%) was the predominant species followed by Cx. tritaeniorhynchus (2.97%) and Cx. gelidus (2.25%). In saline tolerant study, the maximum adult emergence occurred at 5 parts per thousand. The number of larvae surviving till adult stage decreased progressively with increased salinity. The wing size decreased significantly with increasing salinities, except those reared in 5ppt. Interpretation: The present study demonstrates that these three mosquitoes have the ability to oviposit and breed in brackish water. The study also reveals the health risks associated with vector mosquitoes developing and their frequency of abundance in brackish water, particularly in the context of rising sea levels due to global warming.

Bioinformatics characterization of envelope glycoprotein from Kyasanur Forest disease virus

Background & objectives: Kyasanur Forest disease (KFD) is a febrile illness characterized by haemorrhages and caused by KFD virus (KFDV), which belongs to the Flaviviridae family. It is reported to be an endemic disease in Shimoga district of Karnataka State, India, especially in forested and adjoining areas. Several outbreaks have been reported in newer areas, which raised queries regarding the changing nature of structural proteins if any. The objective of the study was to investigate amino acid composition and antigenic variability if any, among the envelope glycoprotein (E-proteins) from old and new strains of KFDV. Methods: Bioinformatic tools and techniques were used to predict B-cell epitopes and three-dimensional structures and to compare envelope glycoprotein (E-proteins) between the old strains of KFDV and those from emerging outbreaks till 2015. Results: The strain from recent outbreak in Thirthahalli, Karnataka State (2014), was similar to the older strain of KFDV (99.2%). Although mutations existed in strains from 2015 in Kerala KFD sequences, these did not alter the epitopes. Interpretation & conclusions: The study revealed that though mutations existed, there were no drastic changes in the structure or antigenicity of the E-proteins from recent outbreaks. Hence, no correlation could be established between the mutations and detection in new geographical areas. It seems that KFDV must be present earlier also in many States and due to availability of testing system and alertness coming into notice now.

Assessment of the efficacy and tolerability of a fixed dose combination of atorvastatin 10 mg + metformin SR 500 mg in diabetic dyslipidaemia in adult Indian patients.

Type 2 diabetes mellitus is associated with a marked increase in the risk of coronary heart disease (CHD) or stroke (by a factor of two to three compared with non-diabetic patients), and cardiovascular disease (CVD) accounts for the majority of deaths among patients with diabetes. A new fixed dose combination containing atorvastatin 10 mg + metformin SR 500 mg is being introduced in the Indian market for the treatment of dyslipidaemia in diabetic patients. The present study was therefore undertaken to assess efficacy, safety and tolerability of a fixed dose combination of atorvastatin 10mg + metformin SR 500mg in adult Indian patients with diabetic dyslipidaemia. The final protocol was approved by relevant ethics committee before the initiation of study. Informed consent was obtained from all the patients prior to enrollment in study. The total duration of study was 14 weeks including two weeks dietary run in period. Patients fulfilling the selection criteria received a single oral tablet of fixed dose combination of atorvastatin 10mg + metformin SR 500mg once daily for 12 weeks. The primary efficacy parameters were assessed by evaluating reduction in fasting and postprandial plasma glucose concentration levels at baseline and thereafter at each follow up visit at 2, 4, 8 and 12 weeks and plasma lipid profile and glycosylated Hb levels at baseline and end of study. The secondary efficacy parameters were assessed by evaluating percentage change from baseline at the end of the study (week 12) in the plasma concentration of the various lipid parameters such as total, HDL-, LDL- and very low density (VLDL)-cholesterol, triglycerides, Apo B, Apo A1, TC/LDL ratio, LDL/ HDL ratio, and percentage of patients achieving LDL-cholesterol goals as per NCEP ATP III guidelines. A total of 213 patients were enrolled in the study. Of these seven patients were lost to follow-up and considered as drop-outs. Therapy with the fixed dose combination of atorvastatin 10 mg + metformin SR 500 mg resulted in a significant reduction in the mean plasma fasting and postprandial glucose levels (35 and 38.8% respectively). There was a steep fall in the HbA1c levels from baseline levels of 8.76% to 6.74% (23.1%). There was also a significant (p < 0.05) reduction in mean total cholesterol (31.2%), LDL cholesterol (35.4%), VLDL-cholesterol (19.6%) and a significant increase HDL-cholesterol (9.5%). Thus there appeared to be trend towards reducing atherosclerosis following therapy with the fixed dose combination of atorvastatin 10 mg + metformin SR 500 mg. Mean body mass index was significantly reduced in the patients in the present study following therapy with the study drugs. The fixed dose combination of atorvastatin with metformin was well tolerated with mostly gastro-intestinal adverse events being reported in the current study. Moreover, most of the adverse events were mild to moderate in intensity and disappeared with continued treatment. In conclusion, the results of the present study suggest that, the fixed dose combination of atorvastatin 10 mg + metformin SR 500 mg is efficacious and well tolerated therapeutic modality in patients with diabetic dyslipidaemia. Furthermore this combination offers dosage convenience to the patient and by virtue of its dual mode of action is a useful addition to the therapeutic armamentarium for patients with diabetic dyslipidaemia.

Risk factors for non-adherence to directly observed treatment (DOT) in a rural tuberculosis unit, South India.

OBJECTIVE: To identify risk factors for non-adherence of tuberculosis (TB) patients to DOT. METHODS: Retrospective study of TB patients by logistic regression analysis to identify risk factors for non-adherence. RESULTS: Of the 1666 patients interviewed, 1108 (67%) adhered and 558 (33%) did not adhere to DOT. Of 558 patients, the risk factors associated with non-adherence were illiteracy (39%), difficulty in accessing health facility (57%), and non-government DOT centre (43%). CONCLUSION: Patients should be educated about tuberculosis and importance of DOT. All DOT centres, including Non-government DOT centres, should be made more accessible and patient-friendly.

Influence of drug susceptibility on treatment outcome and susceptibility profile of failures to category II regimen.

Objective: To assess the influence of drug resistance on treatment outcome among patients treated with Category-II regimen and document drug susceptibility pattern of “Failures” to this regimen. Design: A retrospective analysis of patients registered from May 1999 through December 2004. Results: Treatment success was 42% among 572 patients and was similar among patients with fully susceptible or resistant but non-MDR organisms (41% of 254 and 40% of 128 patients, respectively). Among 49 MDR-TB patients, 27% had successful treatment outcome. The failure rates among patients with fully susceptible, resistant but non-MDR and MDR bacilli, were 6%, 12% and 27% respectively. Default was significantly higher among males (53% vs. 34%: p<0.01) smokers (57% vs. 36%: p <0.001), alcoholics (58% vs. 39%: p <0.001) and patients with higher initial smear grading (2+ or 3+, 56% vs. scanty or 1+, 44%: p <0.01). DST results were available for 60% (31 of 52) of failures and 10 had MDR-TB. Conclusion: The low success rate to the re-treatment regimen was mainly due to non-compliance. Failure was observed among 9% of patients and MDR-TB was 32% among Category II failures. The currently recommended Category II regimen appears to be adequate for majority of re-treatment cases.

Performance of a DOTS programme: Administrative and Technical Challenges - A field report from a district in south India.

Background: Performance of tuberculosis (TB) control programme depends on the functioning of health facilities (HFs). TB control programmes have been evaluated based on the programme indicators of conversion and cure. We have attempted to correlate the programme performance based on the availability of staff and their performance at the HF level. Objective: To correlate the performance of HFs to programme indicators, conversion and cure of patients treated under DOTS, in a district of south India. Design: Analysis of the data on new sputum smear-positive cases registered in 17 HFs during 1999-2003 was undertaken using TB register. The HFs with a low conversion or cure rates were identified and the reasons for the same were analysed. A scoring system was designed for the functioning of the HFs based on staff availability, supervision and review meetings which was correlated with programme performance. Univariate and multivariate analyses were performed. Results: Of 1893 new smear-positive patients registered during the study period, conversion was 1582 (83.6%) with cure rate of 76.4% (1447 of 1893), 254 (13.4%) default, 94 (5.0%) failure and 85 (4.5%) death. The conversion rates increased from 76% in 1999 to 87% in 2003; a statistically significant trend (c2 = 15.9; P<0.001). Similarly, a significant increase in trend (c2 = 4.0; P < 0.05) was observed in cure rates also (71 to 80%). The HFs were broadly classified into four groups namely; poor, fair, good and very good based on functioning scores .Correlation co-efficient was 0.77 between functioning of the HFs and conversion, and 0.76 between functioning and cure (P<0.01). Lack of regular review meetings was found to be independently associated with poor programme performance. Conclusion: Availability of staff such as Medical Officer, Laboratory Technician, and regular supervisory visits and review meetings are essential for a well functioning of programme. There is significant impact on DOTS with good functioning of HFs.

Elderly lady with ascites.

An elderly lady with amylase-rich ascites is presented, whose isoenzyme estimation revealed salivary type amylase. Tumour hyperamylasemia is an important group among the nonpancreatic causes of elevated amylase.

Socio-economic dimensions of tuberculosis control: review of studies over two decades from Tuberculosis Research Center.

Tuberculosis (TB) affects the most productive age group and the resultant economic cost for society is high. Even though diagnostic and treatment services under TB control are offered free of cost, TB patients do incur out of pocket expenditure. Tuberculosis Research Centre under took a series of studies on economic aspects of TB. We interviewed TB patients enrolled under TB control programme in south India, and assessed the following: socio-economic status of patients (SLI - standard of living index) and economic impact on patients, families both before and after Revised TB Control Programme (RNTCP) and the impact of parental TB on children. In addition patient's perceptions of physical, mental, social well being during and after completion of treatment were also elicited. A TB patient incurs out of pocket expenditure of Rs 5986 amounting to about 13,000 crores a year for the country; 11% of children dropped out of school on account of parental illness and 20% of the children had to take up employment in order to supplement income especially if the father had TB. About 64% of TB patients registered under RNTCP were poor (low SLI). The provider cost to treat a TB patient under RNTCP was Rs 1587/- for Category I, Rs 1924/- for Category II and Rs 1417/- Category III. At the end of treatment even though 47% of patients continued to have respiratory symptoms, 54% of patients perceived 'happy mental status'. Majority of the patients registered under RNTCP were poor. Patients' and provider costs and the impact on patients including families on account of TB were enormous. About half of TB patients, despite completing treatment successfully had persistent respiratory ill health resulting in frequenting health facilities. This information is vital for programme planners indicating that the existing control programs have been ineffective. To achieve success in control programs, the existing ones should be amended as there is evolution of resistance in the parasite as well as the vector. Public health education, to make the people aware about preventive aspects of the disease is important. The possibility of the existence of animal reservoirs should also be considered and checked out for better control measures.

Estimating provider cost for treating patients with tuberculosis under Revised National Tuberculosis Control Programme (RNTCP).

Background: The Indian tuberculosis control programme is the second largest health programme in the world. Sustaining this programme in India will require continued financial support, particularly for drugs and contractual personnel. In addition, the costs for diagnosis, supervision and salaries for regular programme personnel need to be sustained. Objective: To measure unit provider cost for treating patients with tuberculosis. Methods: All government health facilities situated in one tuberculosis unit (TU) of Tiruvallur district were visited in order to evaluate daily practice of TB diagnosis and treatment. We interviewed administrators in these health facilities to gather data on modalities for diagnosis, treatment and monitoring of tuberculosis patients. In addition, relevant financial records from all health facilities were scrutinised for data collection. The cost analysis was done for diagnosis, treatment and monitoring of TB patients treated under DOTS programme in the year 2002 For this study only the recurrent cost (not the capital cost) is considered, even though the programme puts in a lot of investment at the preparatory stage of the programme e.g. upgrading of labs and drugs stores, microscopes, motorcycles etc. Cost incurred on smear microscopy, chest X-ray and drugs were classified as direct cost. Indirect cost is calculated based on proportion of staff time for TB care delivery and for supervision of TB services. The exchange rate at the time study was 1$=Rs 46. Results: Unit cost for smear microscopy was estimated to be Rs 10/-; for radiography Rs 25/-; and drug cost for Category I Rs 392/-; Category I with extension Rs 495/-; Category II Rs 729/-; Category II with extension Rs 832/- and Category III Rs 277/-. Including other recurrent expenditures like salary, materials, and maintenance, the overall unit provider cost to treat a TB patient was Rs 1587/- for Category I, Rs 1924/- for Category II and Rs 1417/- Category III. Conclusion: TB inflicts considerable economic burden on the overall health system. This information is vital for policy makers and planners to allocate adequate budget to the programme.

A double-blind study of oral salbutamol supplement and repeat sputum smear microscopy in enhancing diagnosis of smear-negative pulmonary tuberculosis in south India.

Background: As routine culture facilities are not available in TB control programme in low income countries like India, there is an urgent need to improve the sensitivity of sputum microscopy, especially in diagnosis of smear negative pulmonary TB. Methodology: In a double blind placebo controlled study, the role of repeat sputum microscopy after antibiotics and oral salbutamol supplement in improving the diagnosis of smear negative TB suspects was investigated in an urban TB clinic. We undertook culture examinations for all study patients to find out proportions of TB cases in this series. Results: Of 206 enrolled, (101 salbutamol (S), 105 placebo (P) groups) 26 were positive by repeat sputum smear examination; similar in two groups (S 16, P 10, p = 0.25). In all, 40 (S 23, P 17) including 26 smear- positives, were culture -positive for M. tuberculosis. Conclusions: Two thirds of initially smear negative but culture positive TB patients were smear positive on repeat sputum examination. Thus, repeat sputum smear microscopy for TB suspects improved the diagnosis, nevertheless oral salbutamol therapy was not beneficial. In resource poor settings, repeat sputum smear microscopy after a trial of antibiotics, could significantly improve the diagnosis of smear-negative PTB patients.

Near-fatal amlodipine poisoning.

Amlodipine poisoning is very rare and only few cases have been reported in English literature. We report here a case of severe amlodipine poisoning with non-cardiogenic pulmonary edema.

Management of non-pulmonary forms of tuberculosis: review of TRC studies over two decades.

Tuberculosis Research Centre, Chennai has been conducting randomized controlled clinical trials in both adults and children (n = 1015) in various non-pulmonary forms of tuberculosis, for the last two decades. This communication discusses the salient findings of these studies. The important finding is that short course chemotherapeutic regimens have been proven to be highly effective in tuberculosis of spine, superficial tuberculous lymphadenitis, abdominal tuberculosis, brain tuberculoma and Pott's paraplegia. However, in tuberculous meningitis, the outcome appears to be directly related to the stage of the disease on admission. The intermittent regimens have been found to be as effective as daily regimens. The other important aspects highlighted are the need to obtain bacteriological/histo-pathological confirmation by resorting to relevant diagnostic procedures, value of Mantoux as a diagnostic tool and role of surgery.

A case of pre-leukaemia masquerading as haemolytic anaemia due to multi-enzymopathy of the red cell.

A 19 year old woman presented as a case of haemolytic anaemia due to multi-enzyme deficiency of the erythrocyte. After a transient improvement with folic acid therapy, she developed acute myeloblastic leukaemia. This is the second reported case of a myelodysplastic syndrome presenting with a haemolytic picture and subsequently developing an acute myeloblastic leukaemia.

Conformational studies on nucleotide–amino acid interactions leading to origin of life.

Living processes may be defined as the self-sustained chemical reactions based on the special chemical machinery of nucleic acid-directed protein synthesis. Its genesis may be traced to the molecular interaction between nucleotides and amino acids leading to a primitive adaptor-mediated ordered synthesis of polypeptides. A primitive decoding system is described and its characteristics are shown to imitate, in a primitive manner, the present-day elaborate machinery of protein synthesis. This molecular interaction theory may be rightly considered as the missing link between the Protochemical and Biological Evolution. The origin of chiral specificity observed in living organisms is also traced to this specific molecular interaction in the protobiological milieu.