Insecticide susceptibility of Phlebotomus argentipes sandflies, vectors of visceral leishmaniasis in India.

OBJECTIVES: Indoor residual spraying (IRS) with insecticides is the main vector control intervention for the elimination of visceral leishmaniasis in India. After a change in IRS policy in 2015 due to widespread resistance of Phlebotomus argentipes to DDT, IRS with DDT was replaced with alpha-cypermethrin IRS in 2016. The objective of the present study was to evaluate the susceptibility of P. argentipes to DDT and its alternatives, namely malathion and pirimiphos-methyl (organophosphates); alpha-cypermethrin, deltamethrin, lambda-cyhalothrin and permethrin (pyrethroids), and bendiocarb and propoxur (carbamates), in support of visceral leishmaniasis elimination in India. METHODS: Phlebotomus argentipes sandflies were collected from the visceral-leishmaniasis endemic states of Bihar, Jharkhand and West Bengal. In the WHO tube tests, the phenotypic susceptibility of F1, 2-day old, non-blood fed females were determined against filter papers impregnated with DDT 4%, malathion 5%, pirimiphos-methyl 0.25%, alpha-cypermethrin 0.05%, deltamethrin 0.05%, lambda-cyhalothrin 0.05%, permethrin 0.75%, bendiocarb 0.1% and propoxur 0.1%, which were sourced from Universiti Sains Malaysia. The knockdown of sandflies after 1-h exposure and mortality at 24 h after the 1-h exposure period were scored. RESULTS: Mean mortality of P. argentipes 24 h after exposure in tube tests was 22.6% for DDT and ≥ 98% for other insecticide-impregnated papers tested. CONCLUSION: Phlebotomus argentipes continues to be highly resistant to DDT with no reversal of resistance after DDT's withdrawal from IRS. P. argentipes was fully susceptible to pyrethroid, organophosphate and carbamate insecticides tested. Regular monitoring is warranted for insecticide resistance management in sandfly vectors.

"Comparison of Nalbuphine Hydrochloride and Fentanyl as an Adjuvant to Bupivacaine for Spinal Anesthesia in Lower Abdominal Surgeries:" A Randomized, Double-blind Study.

BACKGROUND AND AIMS: Opioids have been favored as adjuvants to local anesthetics during spinal anesthesia. Nalbuphine, a µ-receptor antagonist and ĸ-receptor agonist, seems to be a suitable adjuvant to local anesthetics. The aim of this study was to compare postoperative analgesia and adverse effects of nalbuphine and fentanyl when used as an adjuvant to hyperbaric bupivacaine during spinal anesthesia. MATERIALS AND METHODS: Sixty patients belonging to the American Society of Anesthesiologists Physical Status I and II were randomly allocated into two groups of thirty each. Patients in bupivacaine nalbuphine group (Group BN) received 0.8 mg (0.3 ml) of nalbuphine with 12.5 mg (2.5 ml) of 0.5% hyperbaric bupivacaine diluted to 3 ml and bupivacaine-fentanyl group (Group BF) received 25 µg (0.5 ml) of fentanyl with 12.5 mg (2.5 ml) of 0.5% hyperbaric bupivacaine. Patients were assessed for hemodynamic changes, sensory and motor block, early postoperative analgesia, and adverse effects. RESULTS: Onset, duration of sensory and motor block, and duration of effective analgesia were comparable between both groups. Postoperative visual analog scale score was 4.8 ± 1.12 in Group BN, and in Group BF, it was 3.86 ± 1.04 which was statistically highly significant (P = 0.0007). The number of patients demanding rescue analgesia in early postoperative period was 18 (60.0%) in Group BN and 7 (23.33%) in Group BF which was statistically significant (P = 0.004). CONCLUSION: Fentanyl was more efficient than nalbuphine in providing early postoperative analgesia when used as an adjuvant to hyperbaric bupivacaine.

Measurement of recent exposure to Phlebotomus argentipes, the vector of Indian visceral Leishmaniasis, by using human antibody responses to sand fly saliva.

Antibody (IgG) responses to the saliva of Phlebotomus argentipes were investigated using serum samples from regions of India endemic and non-endemic for visceral leishmaniasis (VL). By pre-adsorbing the sera against the saliva of the competing human-biting but non-VL vector P. papatasi, we significantly improved the specificity of a P. argentipes saliva enzyme-linked immunosorbent assay. Using this method, we observed a statistically significant correlation between antibodies to P. argenitpes saliva and the average indoor density of female sand flies. Additionally, the method was able to detect recent changes in vector exposure when sera from VL patients were assayed before, during, and after hospitalization and protected from sand fly bites under untreated bed nets. Collectively, these results highlight the utility of antibodies to P. argentipes saliva as an important tool to evaluate VL vector control programs.

Phlebotomus argentipes seasonal patterns in India and Nepal.

The current control of Phebotomus argentipes (Annandale and Brunetti), the vector of Leishmania donovani (Laveran and Mesnil), on the Indian subcontinent is base on indoor residual spraying. The efficacy of this method depends, among other factors, on the timing and number of spraying rounds, which depend on the P. argentipes seasonality. To describe P. argentipes' seasonal patterns, six visceral leishmaniasis (VL) endemic villages, three in Muzaffarpur and three in Sunsari districts in India and Nepal, respectively, were selected based on accessibility and VL incidence. Ten houses per cluster with the highest P. argentipes density were monitored monthly for 15-16 mo using Center for Disease Control and Prevention light traps. Minimum and maximum temperature and rainfall data for the months January 2006 through December 2007 were collected from the nearest available weather stations. Backwards stepwise regression was used to generate the minimal adequate model for explaining the monthly variation in P. argentipes populations. The seasonality of P. argentipes is similar in India and Nepal, with two annual density peaks around May and October. Monthly P. argentipes density is positively associated with temperature and negatively associated with rainfall in both study sites. The multivariate climate model explained 57% of the monthly vectorial abundance. Vector control programs against P. argentipes (i.e., indoor residual spraying) should take into account the seasonal described here when implementing and monitoring interventions. Monitoring simple meteorological variables (i.e., temperature, rainfall) may allow prediction of VL epidemics on the Indian subcontinent.

Effect of village-wide use of long-lasting insecticidal nets on visceral Leishmaniasis vectors in India and Nepal: a cluster randomized trial.

BACKGROUND: Visceral leishmaniasis (VL) control in the Indian subcontinent is currently based on case detection and treatment, and on vector control using indoor residual spraying (IRS). The use of long-lasting insecticidal nets (LN) has been postulated as an alternative or complement to IRS. Here we tested the impact of comprehensive distribution of LN on the density of Phlebotomus argentipes in VL-endemic villages. METHODS: A cluster-randomized controlled trial with household P. argentipes density as outcome was designed. Twelve clusters from an ongoing LN clinical trial--three intervention and three control clusters in both India and Nepal--were selected on the basis of accessibility and VL incidence. Ten houses per cluster selected on the basis of high pre-intervention P. argentipes density were monitored monthly for 12 months after distribution of LN using CDC light traps (LT) and mouth aspiration methods. Ten cattle sheds per cluster were also monitored by aspiration. FINDINGS: A random effect linear regression model showed that the cluster-wide distribution of LNs significantly reduced the P. argentipes density/house by 24.9% (95% CI 1.80%-42.5%) as measured by means of LTs. INTERPRETATION: The ongoing clinical trial, designed to measure the impact of LNs on VL incidence, will confirm whether LNs should be adopted as a control strategy in the regional VL elimination programs. The entomological evidence described here provides some evidence that LNs could be usefully deployed as part of the VL control program. TRIAL REGISTRATION: ClinicalTrials.gov CT-2005-015374.

Post-transcriptional regulation of miR-27 in murine cytomegalovirus infection.

In mammals, microRNAs (miRNAs) can play diverse roles in viral infection through their capacity to regulate both host and viral genes. Recent reports have demonstrated that specific miRNAs change in expression level upon infection and can impact viral production and infectivity. It is clear that miRNAs are an integral component of viral-host interactions, and it is likely that both host and virus contain mechanisms to regulate miRNA expression and/or activity. To date, little is known about the mechanisms by which miRNAs are regulated in viral infection. Here we report the rapid down-regulation of miR-27a in multiple mouse cell lines as well as primary macrophages upon infection with the murine cytomegalovirus. Down-regulation of miR-27a occurs independently from two other miRNAs, miR-23a and miR-24, located within the same genomic cluster, and analysis of pri-miRNA levels suggest that regulation occurs post-transcriptionally. miR-27b, a close homolog of miR-27a (20/21 nucleotide identity), also decreases upon infection, and we demonstrate that both miR-27a and miR-27b exert an antiviral function upon over-expression. Drug sensitivity experiments suggest that virus entry is not sufficient to induce the down-regulation of miR-27 and that the mechanism requires synthesis of RNA. Altogether, our findings indicate that miR-27a and miR-27b have antiviral activity against MCMV, and that either the virus or the host encodes molecule(s) for regulating miR-27 accumulation, most likely by inducing the rapid decay of the mature species.

Long-lasting insecticidal nets fail at household level to reduce abundance of sandfly vector Phlebotomus argentipes in treated houses in Bihar (India).

OBJECTIVE: To determine whether the use of long-lasting insecticidal nets (LLINS) at household level are effective in reducing the abundance of Phlebotomus argentipes, vector of anthroponotic visceral leishmaniasis in India, Nepal and Bangladesh. METHODS: The impact of two long-lasting nets (Olyset and PermaNet) on indoor sandfly abundance was evaluated in selected houses of three endemic hamlets in Bihar (India). It was assumed that most sandflies breed inside the houses and that LLINs would progressively reduce the indoor density during the reproduction season. A campaign of indoor spraying with dichloro-diphenyl-trichloroethane (DDT) interfered with the trial but did not affect the sandfly population. Results Only the density of males of P. argentipes was significantly reduced by both the LLINs but not the females. CONCLUSIONS: These findings suggest that most female sandflies are coming from outside and that LLINs do not reduce their entry rate.

Discrete clusters of virus-encoded micrornas are associated with complementary strands of the genome and the 7.2-kilobase stable intron in murine cytomegalovirus.

The prevalence and importance of microRNAs (miRNAs) in viral infection are increasingly relevant. Eleven miRNAs were previously identified in human cytomegalovirus (HCMV); however, miRNA content in murine CMV (MCMV), which serves as an important in vivo model for CMV infection, has not previously been examined. We have cloned and characterized 17 novel miRNAs that originate from at least 12 precursor miRNAs in MCMV and are not homologous to HCMV miRNAs. In parallel, we applied a computational analysis, using a support vector machine approach, to identify potential precursor miRNAs in MCMV. Four of the top 10 predicted precursor sequences were cloned in this study, and the combination of computational and cloning analysis demonstrates that MCMV has the capacity to encode miRNAs clustered throughout the genome. On the basis of drug sensitivity experiments for resolving the kinetic class of expression, we show that the MCMV miRNAs are both early and late gene products. Notably, the MCMV miRNAs occur on complementary strands of the genome in specific regions, a feature which has not previously been observed for viral miRNAs. One cluster of miRNAs occurs in close proximity to the 5' splice site of the previously identified 7.2-kb stable intron, implying a variety of potential regulatory mechanisms for MCMV miRNAs.

Characterization of ionically bound peroxidases from apple (Mallus pumilus) fruits.

Ionically bound peroxidases (POD) were salt extracted from the pulp of four Indian apple varieties, i.e., Golden delicious HP, Golden delicious JK, Red delicious, and Royal delicious. They were precipitated with chilled ethanol. Thermal treatments of partially purified enzymes were given from 40-70 degrees C for 30 minutes. Golden delicious HP peroxidase showed thermostability at 60 degrees C, while three other peroxidases were observed at 50 degrees C. Phenolic compounds (i.e., caffeic acid, ferulic acids, p-coumaric acid, protocatechuic acid) and metal ions (i.e., Cu2+ and Fe2+) activated all apple peroxidases. However, Mn2+ inhibited the peroxidases from Golden delicious HP, Golden delicious JK, and Red delicious, and a substantial increase was observed in Royal delicious peroxidase. Mg2+ inhibited the peroxidases from Golden delicious HP and Red delicious, but marginal activation was reported in peroxidases from Golden delicious JK and Royal delicious. Zn2+ established stimulation in Golden delicious HP and Golden delicious JK peroxidases, but inhibition was observed in peroxidases in Red delicious and Royal delicious.. Methionine, proline, tryptophan, and valine stimulated all four apple peroxidases, but cysteine showed inhibition in Golden delicious JK.

Role of tranexamic acid in management of dysfunctional uterine bleeding in comparison with medroxyprogesterone acetate.

Currently, tranexamic acid (TXA) is used as 4 g/day in menorrhagia This prospective randomised study included 100 cases to assess efficacy and safety of 2 g/day TXA in dysfunctional uterine bleeding (DUB) vs cyclical 10 mg twice-daily medroxyprogesterone acetate (MPA) for 3 cycles. Follow-ups were made monthly for 3 months during therapy, then 3 months after. Mean pictorial blood loss assessment chart (PBAC) score decreased from 356.9 to 141.6 in the TXA group and from the pre-treatment 370.9 to 156.6 with MPA and mean reduction of blood loss was 60.3% with TXA and 57.7% with MPA after 3 months (p < 0.005 in both groups). Lack of response during treatment was seen in three patients (6.1%) TXA and in 13 patients (28.9%) with MPA (p = 0.003). In patients who reported 3 months after stopping the treatment, 66.7% in TXA group and 50% in MPA had recurrence of menorrhagia, (p = 0.155). During the 6 months study period more hysterectomies were performed in the MPA than in the TXA group (17.8% vs 4%; p = 0.002). We conclude that TXA in 2 g/day dosage is an effective and safe option in DUB.

Aflatoxin B1 contamination in wheat grain samples collected from different geographical regions of India: A multicenter study.

In a multicenter study conducted by the Indian Council of Medical Research, 1,646 samples of wheat grain collected from rural and urban areas of 10 states representing different geographical regions of India were analyzed for aflatoxin B1 (AFB1). AFB1 concentrations of > or = 5 microg kg(-1) were recorded in 40.3% of the samples, and concentrations above the Indian permissible regulatory limit of 30 microg kg(-1) were found in 16% of the samples. The proportion of samples with AFB1 concentrations above the Indian regulatory limit ranged from 1.7 to 55.8% in different states, with the minimum in Haryana and the maximum in Orissa. The variation in wheat contamination among states seems to be mainly the result of unsatisfactory storage conditions. Median AFB1 concentrations of 11, 18, and 32 microg kg(-1) were observed in samples from Uttar Pradesh, Assam, and Orissa, respectively; concentrations in other states were <5 microg kg(-1). The maximum AFB1 concentration of 606 microg kg(-1) was observed in a sample from the state of Uttar Pradesh. The calculated probable daily intakes of AFB1 through consumption of contaminated wheat for the population in some states were much higher than the suggested provisional maximum tolerable daily intake. Human health hazards associated with such AFB1 exposure over time cannot be ruled out.

Residues of DDT and HCH in wheat samples collected from different states of India and their dietary exposure: A multicentre study.

Under a multicentre study conducted by the Indian Council of Medical Research, 1712 samples of wheat grain/flour were collected from urban and rural areas in 11 states representing different geographical regions of India. These samples were analysed for residues of DDT (2,2-bis (p-chlorophenyl)-1,1,1-trichloro ethane) and different isomers of HCH (1,2,3,4,5,6-hexachloro cyclohexane, a mixture of isomers) by gas-liquid chromatography. Residues of DDT were detected in 59.4% of 1080 samples of wheat grain and in 78.2% of 632 samples of wheat flour. Different isomers of HCH were present in about 45-80% of the samples of wheat grain/flour. Medians of DDT and total HCH, respectively, for pooled samples of wheat grain were 0.013 and 0.035 mg kg(-1), while those for wheat flour were 0.01 and 0.02 mg kg(-1). Estimated daily intakes of DDT and different isomers of HCH through the consumption of wheat contaminated at their median and 90th percentiles constituted a small proportion of their acceptable daily intakes. Amongst the pesticide residues analysed, statutory maximum residue limits have been fixed only for gamma-HCH in wheat in India, as 0.1 mg kg(-1) in wheat grain and zero in wheat flour. Residue levels of gamma-HCH exceeded these maximum residue limits in five of 1080 samples of wheat grain and in 340 of 632 samples of wheat flour. The failure to meet the requirement of the gamma-HCH maximum residue limit in large number of wheat flour samples was attributed to the fixation of practically unachievable zero limit. Comparing the previous studies and the present one, the levels of residues of DDT and HCH in wheat were significantly decreased.

Aflatoxin B(1) contamination of parboiled rice samples collected from different states of India: A multi-centre study.

Under a multi-centre study conducted by the Indian Council of Medical Research, 1,511 samples of parboiled rice were collected from rural and urban areas of 11 states representing different geographical regions of India. These samples were analysed for contamination with aflatoxin B(1.) The presence of aflatoxin B(1) at levels=5 microg g(-1) was found in 38.5% of the total number of samples of the parboiled rice. About 17% of the total samples showed the presence of aflatoxin B(1) above the Indian regulatory limit of 30 microg kg(-1). No statistically significant difference in percentage of samples contaminated with >30 microg kg(-1) was observed between pooled rural (19.4%) and urban (14.5%) data. A median value of 15 microg kg(-1) of aflatoxin B(1) was observed in samples from Assam, Bihar and Tripura. In all other states surveyed the median value was <5 microg?kg(-1).

Residues of DDT and HCH pesticides in rice samples from different geographical regions of India: a multicentre study.

As part of a multicentre study conducted by the Indian Council of Medical Research, 2000 samples of rice were collected from rural and urban areas of 13 states representing different geographical regions of India. The samples were analyzed for residues of DDT [2,2-bis(p-chlorophenyl)-1,1,1-trichloroethane] and different isomers of HCH [1,2,3,4,5,6-hexachlorocyclohexane, a mixture of isomers] by gas-chromatography. Residues of DDT and HCH, respectively, were detected in about 58 and 73% of the samples analysed. Medians of both DDT and HCH in rice samples were around 0.01 mg kg (-1). Concentrations of alpha-, beta-, gamma- and delta-HCH exceeded a maximum residue limit of 0.05 mg kg (-1) for each isomer in rice fixed by the Ministry of Health and Family Welfare of the Indian Government in 4.3, 2.6, 1.7 and 1.2% of the samples, respectively. There is no statutory limit fixed in India for DDT residues in cereals. Its maximum residue limit of 0.1 mg kg (-1) in cereals recommended by Codex was exceeded by about 2% of the samples examined. Estimated intake of DDT and isomers of HCH through consumption of rice contaminated at their median and 90th percentiles constituted a small proportion of their acceptable daily intakes.