Ecosystem health assessment of East Kolkata Wetlands, India: Implications for environmental sustainability.

The East Kolkata Wetlands (EKW) in Kolkata, India, span 12,500 ha and are a vital ecological zone providing several benefits, including water purification, flood control, and biodiversity support. This study investigated land use and land cover (LULC) alterations in the EKW from 1991 to 2023, using a random forest (RF) machine learning model. Significant LULC changes were observed over the 32 years, with wetland areas decreasing from 91.2 km2 in 1991 to 33.4 km2 in 2023, reflecting substantial habitat loss and reduced ecosystem services. Conversely, agricultural land expanded from 27.8 km2 to 58.7 km2, driven by economic and food production needs, and built-up areas increased dramatically from 0.2 km2 to 10.5 km2, indicating rapid urbanization. This study evaluated the health, resilience, and ecosystem functionality of EKW by analysing human-induced land use changes and using ecological indicators and landscape metrics. Landscape and class level metrics such as PLAND, largest patch index (LPI), total edge (TE), edge density (ED), number of patches (NP), and patch density (PD) were used to analyse the spatiotemporal dynamics of the wetlands. This study revealed a significant increase in fragmentation, with the number of patches increasing from 2689 in 1991 to 4532 in 2023, despite a consistent decrease in core wetland areas. Ecosystem health indicators, such as the ecosystem structure index (ESI) and landscape deviation degree (LDD), were used to assess landscape metrics and fragmentation changes. The ESI and other metrics revealed significant temporal fluctuations, providing insights into landscape structure, connectivity, and heterogeneity. The ESI improved from 0.87 in 1991 to 1.03 in 2023, indicating enhanced connectivity and diversity. Conversely, the LDD increased from 20.6% to 56.85%, indicating a shift towards impervious surfaces. The vegetation productivity and ecosystem health index (EHI) decreased, indicating biodiversity loss and reduced carbon sequestration. The EHI also dropped from 0.67 to 0.55, signifying ongoing environmental stress. This study emphasizes the need for conservation efforts to maintain the ecological integrity of the EKW amidst urbanization and land use changes and recommends a balanced approach for sustainable urban development and enhanced wetland resilience.

Subnational estimates of life expectancy at birth in India: evidence from NFHS and SRS data.

BACKGROUND: Mortality estimates at the subnational level are of urgent need in India for the formulation of policies and programmes at the district level. This is the first-ever study which used survey data for the estimation of life expectancy at birth ([Formula: see text]) for the 640 districts from NFHS-4 (2015-16) and 707 districts from NFHS-5 (2019-21) for the total, male and female population in India. METHODS: This study calculated annual age-specific mortality rates from NFHS-4 and NFHS-5 for India and all 36 states for the total, male and female population. This paper constructed the abridged life tables and estimated life expectancy at birth [Formula: see text] and further estimated the model parameters for all 36 states. This study linked state-specific parameters to the respective districts for the estimation of life expectancy at birth [Formula: see text]for 640 districts from NFHS-4 and 707 districts from NFHS-5 for the total, male and female population in India. RESULTS: Findings at the state level showed that there were similarities between the estimated and calculated [Formula: see text] in most of the states. The results of this article observed that the highest [Formula: see text] varies in the ranges of 70 to 90 years among the districts of the southern region. [Formula: see text] falls below 70 years among most of the central and eastern region districts. In the northern region districts [Formula: see text] lies in the range of 70 years to 75 years. The estimates of life expectancy at birth [Formula: see text] shows the noticeable variations at the state and district levels for the person, male, and female populations from the NFHS (2015-16) and NFHS (2019-21). In the absence of age-specific mortality data at the district level in India, this study used the indirect estimation method of relating state-specific model parameters with the IMR of their respective districts and estimated [Formula: see text] across the 640 districts from NFHS-4 (2015-16) and 707 districts from NFHS-5 (2019-21). The findings of this study have similarities with the state-level estimations of [Formula: see text] from both data sources of SRS and NFHS and found the highest [Formula: see text] in the southern region and the lowest [Formula: see text] in the eastern and central region districts. CONCLUSIONS: In the lack of [Formula: see text] estimates at the district level in India, this study could be beneficial in providing timely life expectancy estimates from the survey data. The findings clearly shows variations in the district level [Formula: see text]. The districts from the southern region show the highest [Formula: see text] and districts from the central and eastern region has lower [Formula: see text]. Females have higher [Formula: see text] as compared to the male population in most of the districts in India.

Impact of COVID-19 on subnational variations in life expectancy and life disparity at birth in India: evidence from NFHS and SRS data.

BACKGROUND: Measuring life expectancy and life disparity can assist in comprehending how the COVID-19 pandemic has affected the mortality estimates in the Indian population. The present study aims to study the life expectancy and life disparity at birth at the national and subnational levels before and during the COVID-19 pandemic using the NFHS and SRS data. METHODS: The measures Life expectancy at birth ([Formula: see text]) and Life disparity at birth ([Formula: see text]) were computed for the non-pandemic and pandemic years from NFHS (2015-16), SRS (2015) and NFHS (2019-21), SRS (2020) respectively at the national and Subnational level in India. Using NFHS data for the 36 states and SRS data for the 22 states, the study calculates [Formula: see text] and [Formula: see text] by total, male and female population. RESULTS: The [Formula: see text] for male and female decline from 64.3 years and 69.2 years in 2015-16 to 62.9 years and 68.9 years in 2019-21. The [Formula: see text] shows a drop of approximately 1.4 years for males and 0.3 years for females in the pandemic year 2019-21 when compared to the non-pandemic year 2015-16. At the subnational level [Formula: see text] shows a decline for 22 states in person, 23 states in males and 21 states in females in the pandemic year 2019-21 as compared to the non-pandemic years 2015-16. The [Formula: see text] shows a increase for 21 states in person, 24 states in females and 17 states in males in the pandemic year than non-pandemic year. The findings shows a significant losses in [Formula: see text] and gains in [Formula: see text] for males than females in the pandemic year as compared to the non-pandemic year at the subnational level in India. CONCLUSIONS: COVID-19 pandemic has decreased [Formula: see text] and increased [Formula: see text] in the pandemic year 2019-21 at the national and subnational level in India. COVID-19 had a significant impact on the age pattern of mortality for many states and male, female population and delayed the mortality transition in India.

Machine Learning Algorithms for understanding the determinants of under-five Mortality.

BACKGROUND: Under-five mortality is a matter of serious concern for child health as well as the social development of any country. The paper aimed to find the accuracy of machine learning models in predicting under-five mortality and identify the most significant factors associated with under-five mortality. METHOD: The data was taken from the National Family Health Survey (NFHS-IV) of Uttar Pradesh. First, we used multivariate logistic regression due to its capability for predicting the important factors, then we used machine learning techniques such as decision tree, random forest, Naïve Bayes, K- nearest neighbor (KNN), logistic regression, support vector machine (SVM), neural network, and ridge classifier. Each model's accuracy was checked by a confusion matrix, accuracy, precision, recall, F1 score, Cohen's Kappa, and area under the receiver operating characteristics curve (AUROC). Information gain rank was used to find the important factors for under-five mortality. Data analysis was performed using, STATA-16.0, Python 3.3, and IBM SPSS Statistics for Windows, Version 27.0 software. RESULT: By applying the machine learning models, results showed that the neural network model was the best predictive model for under-five mortality when compared with other predictive models, with model accuracy of (95.29% to 95.96%), recall (71.51% to 81.03%), precision (36.64% to 51.83%), F1 score (50.46% to 62.68%), Cohen's Kappa value (0.48 to 0.60), AUROC range (93.51% to 96.22%) and precision-recall curve range (99.52% to 99.73%). The neural network was the most efficient model, but logistic regression also shows well for predicting under-five mortality with accuracy (94% to 95%)., AUROC range (93.4% to 94.8%), and precision-recall curve (99.5% to 99.6%). The number of living children, survival time, wealth index, child size at birth, birth in the last five years, the total number of children ever born, mother's education level, and birth order were identified as important factors influencing under-five mortality. CONCLUSION: The neural network model was a better predictive model compared to other machine learning models in predicting under-five mortality, but logistic regression analysis also shows good results. These models may be helpful for the analysis of high-dimensional data for health research.

Association between physical limitations and depressive symptoms among Indian elderly: marital status as a moderator.

BACKGROUND: Depression among the elderly is well-documented and associated with socio-economic factors, physical and mental health conditions. Few studies have focused on older adults' physical limitations and depressive symptoms. However, very little is known about marital status' role in such associations, especially in India. The present study examines the association between physical limitations and self-reported depressive symptoms and moderating role of marital status in such association separately for men and women. METHODS: The present study used data from the Longitudinal Ageing Study in India (LASI) wave 1, 2017-2018, a nationally and state representative longitudinal large-scale survey of ageing and health. For the present research, a total sample of 20,806 older adults aged 60+ years was selected after excluding missing values. Along with descriptive statistics, binary logistic regression analysis and interaction effect of marital status were applied to examine the association between physical limitations (functional limitations and mobility difficulty) with the depressive symptoms separately for men and women. RESULTS: About 58, 50, and 45% elderly reported having depressive symptoms and had difficulty in 2+ ADLs, 2+ IADLs, and 2+ mobility difficulties, respectively. By the marital status, the prevalence of depressive symptoms was higher among currently unmarried than currently married, irrespective of type and number of physical limitations. The unadjusted, marital and multivariate-adjusted association suggested that elderly with more than two ADLs, IADLs, and mobility difficulty had higher odds of depressive symptoms. The gender stratified interaction effect of marital status and physical limitations on depressive symptoms indicated that currently unmarried elderly, particularly unmarried older women with 2+ ADLs (OR = 2.85; CI 95% = 1.88-3.09), 2+ IADLs (OR = 2.01; CI 95% = 1.74-2.31) and 2+ mobility difficulty (OR = 2.20; CI 95% = 1.86-2.60) had higher odds of depressive symptoms. However, such association was only valid for unmarried men having mobility difficulty. CONCLUSION: The study highlights that the elderly with physical limitations such as ADLs, IADLs, and mobility difficulty require attention and care. Although married elderly are less likely to have depressive symptoms even with all the mentioned physical limitations, unmarried women are more vulnerable to have depressive symptoms with physical limitations.

Impact of COVID-19 on life expectancy at birth in India: a decomposition analysis.

BACKGROUND: Quantifying excess deaths and their impact on life expectancy at birth (e0) provide a more comprehensive understanding of the burden of coronavirus disease of 2019 (COVID-19) on mortality. The study aims to comprehend the repercussions of the burden of COVID-19 disease on the life expectancy at birth and inequality in age at death in India. METHODS: The mortality schedule of COVID-19 disease in the pandemic year 2020 was considered one of the causes of death in the category of other infectious diseases in addition to other 21 causes of death in the non-pandemic year 2019 in the Global Burden of Disease (GBD) data. The measures e0 and Gini coefficient at age zero (G0) and then sex differences in e0 and G0 over time were analysed by assessing the age-specific contributions based on the application of decomposition analyses in the entire period of 2010-2020. RESULTS: The e0 for men and women decline from 69.5 and 72.0 years in 2019 to 67.5 and 69.8 years, respectively, in 2020. The e0 shows a drop of approximately 2.0 years in 2020 when compared to 2019. The sex differences in e0 and G0 are negatively skewed towards men. The trends in e0 and G0 value reveal that its value in 2020 is comparable to that in the early 2010s. The age group of 35-79 years showed a remarkable negative contribution to Δe0 and ΔG0. By causes of death, the COVID-19 disease has contributed - 1.5 and - 9.5%, respectively, whereas cardiovascular diseases contributed the largest value of was 44.6 and 45.9%, respectively, to sex differences in e0 and G0 in 2020. The outcomes reveal a significant impact of excess deaths caused by the COVID-19 disease on mortality patterns. CONCLUSIONS: The COVID-19 pandemic has negative repercussions on e0 and G0 in the pandemic year 2020. It has severely affected the distribution of age at death in India, resulting in widening the sex differences in e0 and G0. The COVID-19 disease demonstrates its potential to cancel the gains of six to eight years in e0 and five years in G0 and has slowed the mortality transition in India.

Molecular characterization of SlATG18f in response to Tomato leaf curl New Delhi virus infection in tomato and development of a CAPS marker for leaf curl disease tolerance.

KEY MESSAGE: Analysis of autophagy-related genes in tomato shows the involvement of SlATG18f in leaf curl disease tolerance and a CAPS marker developed from this gene demonstrates its usefulness in marker-assisted selection. Autophagy is a highly conserved catabolic process regulating cellular homeostasis and adaptation to different biotic and abiotic stress. Several autophagy-related proteins (ATGs) are reported to be involved in autophagic processes, and considering their importance in regulating growth and stress adaptation, these proteins have been identified and characterized in several plant species. However, there is no information available on the role of autophagy-related proteins regulating the tolerance of tomato to tomato leaf curl disease (ToLCD). Given this, the present genome-wide study identified thirty ATG-encoding genes (SlATG) in tomato, followed by their functional characterization. Expression profiling of the SlATG genes in contrasting tomato cultivars subjected to virus infection showed a 4.5-fold upregulation of SlATG18f in the tolerant cultivar. Further, virus-induced gene silencing of SlATG18f in the tolerant cultivar conferred disease susceptibility, which suggested the role of this gene in Tomato leaf curl New Delhi virus tolerance. Comparison of the gene sequence of both tolerant and susceptible cultivars along with the 5' upstream regions identified an SNP (A/T) at -2916 upstream of the start codon. A cleaved amplified polymorphic sequence (CAPS) marker was developed targeting this region, which showed a significant association with the tolerance characteristics in the tomato germplasm (R2 = 0.1787). Altogether, the study identified a potential gene that could be used to develop ToLCNDV tolerant tomato cultivars using transgene-based or marker-assisted breeding-based approaches.

Disaggregated analysis of birth averted due to family planning use in India: An evidence from NFHS-4 (2015-16).

BACKGROUND AND OBJECTIVE: India contributes a major share of global unintended births. It is established that contraception plays a significant role in preventing unintended pregnancies, maternal mortality and induced abortion. In this study, to analyze the effectiveness of our family welfare program, we tried to give district-level estimates of number of births averted due to contraception. DATA AND METHODS: Data for this study came from the cross-sectional, population-based data from the fourth round of National Family Health Survey (NFHS-4) conducted in 2015-16. Here, we discussed two methods based on robust regression for computing number of births averted at district level. Further, we analyzed the percentage increase in births (PIB) that would be experienced by each district in the absence of contraception. RESULTS: Findings of this study clearly showed that there was a huge variation in the estimates of number of births averted among different districts as well as states of India. Out of 640 districts, 315 districts achieved below-replacement fertility and 365 districts have contraceptive prevalence rate (CPR) more than 50 percent. Method 1 found around 22 percent districts showed less than 15 percent reduction in births while Method 2 suggested nearly 14 percent districts predominantly located in Uttar Pradesh, Bihar, Jharkhand, Arunachal Pradesh, Meghalaya and Manipur exhibited less than 30 percent reduction of births due to use of all forms of contraception. At all India level, an average estimate obtained by two methods, nearly 63 million births would have been averted by the use of contraception or 40 percent more than the number of births occurred during that period. CONCLUSION: The study successfully identified the districts that were not performing well at the front of utilization of various family planning methods for birth control. To achieve objectives of National Population Policy (2000), poor-performing districts must be monitored like the government keeps monitoring of Aspirational districts.

Reciprocal changes in CD11c+CD11b+ and CD11c+CD8α+ dendritic cell subsets determine protective or permissive immune response in murine experimental VL.

CD11c+CD8α+ and CD11c+CD11b+ dendritic cells are two major subsets of murine splenic CD11c+ DCs which play a crucial role in T cell priming and shaping Th1/Th2 responses, but their role in the context of experimental visceral leishmaniasis (VL) is poorly understood. Herein, we showed that L. donovani infection in Balb/c mice preferentially decreased the population abundance of CD11c+CD11b+ DCs and increased relative abundance of splenic CD11c+CD8α +DCs. During infection, splenic CD11c+CD11b+ DCs induced Th1 differentiation whereas CD11c+CD8α+ DCs promoted Th2 differentiation. Additionally, treatment of infected mice with miltefosine as experimental control exhibited host defense allowing the restoration of CD11c+CD11b+ population and decrease in CD11c+CD8α+ subset. Furthermore, reciprocal regulation of immune accessory surface molecules, Sema4A and OX40L critically determined Th1/Th2 response induced by these DC subsets during VL. L. donovani infection significantly induced OX40L expression and slightly downregulated SEMA 4A expression in CD11c+CD8α+ DCs whereas miltefosine treatment significantly downregulated OX40L expression along with pronounced upregulation of SEMA 4A expression in CD11c+CD11b+ DCs. SiRNA mediated knockdown of SEMA 4A markedly reduced CD11c+CD11b+ driven IFN-γ, TNF-α and IL-12 synthesis in miltefosine treated mice whereas functional blocking of OX40L decreased CD11c+CD8α+ induced IL-10, IL-4 and TGF-ß synthesis in L. donovani infected group. Vaccination of Balb/c mice with antigen-pulsed + CpG-ODN-activated DC subsets revealed that only antigen-pulsed CD11c+CD11b+ DCs eliminated parasite load in visceral organ and restored protective Th1 cytokine response. Collectively, our results suggest that differential regulation of splenic CD11c+ subsets by L. donovani is essential for disease progression and specific subtypes may be exploited as prophylactic measures against visceral leishmaniasis.

Dandy-Walker variant with precocious puberty: a rare association.

Precocious puberty is characterised by premature appearance of secondary sexual characteristics before the age of 7 years in girls and 9 years in boys. Dandy-Walker malformation comprises a spectrum of intracranial malformations of the posterior fossa. We present a case of a 7-year-old male child who has presented with features of central precocious puberty and on further evaluation has been found to have Dandy-Walker variant and secondary hypothyroidism. The following case report describes this association which is extremely rare and has never been described in literature.

Dulaglutide-induced cerebral venous thrombosis in a patient with type 2 diabetes mellitus.

Though patients with diabetes mellitus are at a high risk of atherothrombotic events, every such event should not be attributed to the disease itself. We present a case of a patient with diabetes with headache and blurring of vision for 3 days. Brain imaging revealed right transverse sinus thrombosis and acute infarct of the right posterior parieto-occipital region, predominantly in the posterior cortical watershed zone. The patient was on subcutaneous dulaglutide for 3 weeks and was having nausea and vomiting. Various causes of cerebral venous thrombosis were ruled out with appropriate laboratory investigations. Finally, cerebral venous thrombosis was attributed to dulaglutide-induced nausea and vomiting which led to severe dehydration.

Leishmania donovani molecules recognized by sera of filaria infected host facilitate filarial infection.

We earlier found that F6 fraction of human filaria Brugia malayi cross-reacted with sera of Leishmania donovani infected hamsters and immunization with F6 inhibited both filarial and leishmanial infections. In the present study, we identified a 52.9-93.6 kDa fraction (Ld1) of L. donovani that cross-reacted with sera of B. malayi infected animals and investigated effect of Ld1 on filarial infection. Immunization of BALB/c mice with Ld1 facilitated B. malayi infection with remarkable increase in parasite burden. Facilitation of filarial infection was associated with downregulated cell proliferation, IL-5, IL-13, IFN-γ, TNF-α, and IL-2 levels and upregulated IL-4 and TGF-ß. Ld1 exposure also suppressed MHC class-I, MHC class-II, and FcεR1 expression, and phagocytosis in naive mouse macrophages, and CD4+, CD8+, and CD19+ cell population in mouse spleen. Two-dimensional electrophoresis and matrix-assisted laser desorption ionization-time of flight-mass spectrometry revealed eight proteins in Ld1: putative heat shock protein (HSP) 70-related protein 1, HSP70 mitochondrial precursor, alanine aminotransferase, 2,3-bisphosphoglycerate-independent phosphoglycerate mutase, protein disulfide isomerase, putative ATPase beta subunit, trypanothione reductase, and a hypothetical protein. HSP70 protein mitochondrial precursor and trypanothione reductase showed homology with Trypanosoma cruzi and L. donovani, respectively, and the rest 6 proteins including hypothetical protein bear homology with L. infantum. In conclusion, the present study for the first time shows that immunization with filarial cross-reactive Ld1 fraction of L. donovani facilitates filarial infection by modulating Th1 and Th2 responses. Ld1 molecules may therefore facilitate filarial infection in filaria-leishmania co-infection.

Antileishmanial Activity of Pyrazolopyridine Derivatives and Their Potential as an Adjunct Therapy with Miltefosine.

A series of pyrazolo(dihydro)pyridines was synthesized and evaluated for antileishmanial efficacy against experimental visceral leishmaniasis (VL). Among all compounds, 6d and 6j exhibited better activity than miltefosine against intracellular amastigotes. Compound 6j (50 mg/kg/day) was further studied against Leishmania donovani/BALB/c mice via the intraperitoneal route for 5 days and displayed >91 and >93% clearance of splenic and liver parasitic burden, respectively. Combination treatment of 6j with a subcurative dose of miltefosine (5 mg/kg) in BALB/c mice almost completely ameliorated the disease (>97% inhibition) by augmenting nitric oxide generation and shifting the immune response toward Th1. Furthermore, investigating the effect of 6j on Leishmania promastigotes revealed that it induced molecular events, such as a loss in mitochondrial membrane potential, externalization of phosphatidylserine, and DNA fragmentation, that ultimately resulted in the programmed cell death of the parasite. These results along with pharmacokinetic studies suggest that 6j could be a promising lead for treating VL as an adjunct therapy with miltefosine.

15d-Prostaglandin J2 induced reactive oxygen species-mediated apoptosis during experimental visceral leishmaniasis.

UNLABELLED: 15-Deoxy-delta (12,14)-prostaglandin J2 (15d-PgJ2) is a potent bioactive lipid mediator, known to possess several roles in cell regulation and differentiation along with antimicrobial efficacy against different bacterial and viral infections. In the present study, we investigated the therapeutic efficacy and mechanism of action of 15d-PgJ2 in vitro in Leishmania donovani promastigotes and infected J774 macrophages, and in vivo in Balb/c mice/golden hamster model of experimental visceral leishmaniasis. 15d-PgJ2 effectively killed L. donovani promastigotes and amastigotes in vitro with IC50 of 104.6 and 80.09 nM, respectively. At 2 mg/kg (mice) and 4 mg/kg (hamster) doses, 15d-PgJ2 decreased >90 % spleen and liver parasite burden. It significantly reduced interleukin (IL)-10 and transforming growth factor (TGF)-ß synthesis in infected macrophages and splenocytes. 15d-PgJ2 induced reactive oxygen species (ROS)-dependent apoptosis of promastigotes by triggering phosphatidyl serine externalization, mitochondrial membrane damage and inducing caspase-like activity. In vitro drug interaction studies revealed an indifference to the synergistic association of 15d-PgJ2 with Miltefosine and Amphotericin-B (Amp-B). Moreover, when combined with sub-curative doses of Miltefosine and Amphotericin-B, 15d-PgJ2 resulted in >95 % parasite removal. Our results suggested that 15d-PgJ2 induces mitochondria-dependent apoptosis of L. donovani and is a good therapeutic candidate for adjunct therapy against experimental visceral leishmaniasis. KEY MESSAGE: 15d-PgJ2 effectively eliminated both promastigotes and amastigotes form of L. donovani. 15d-PgJ2 decreased parasite burden from infected mice and hamsters with reduced Th2 cytokines. 15d-PgJ2 induced ROS-mediated mitochondrial apoptosis of L. donovani promastigotes. 15d-PgJ2 is a good therapeutic candidate for adjunct therapy with Miltefosine and Amp-B.

Combination of liposomal CpG oligodeoxynucleotide 2006 and miltefosine induces strong cell-mediated immunity during experimental visceral leishmaniasis.

Immuno-modulators in combination with antileishmanial drug miltefosine is a better therapeutic approach for treatment of Visceral Leishmaniasis (VL) as it not only reduces the dose of miltefosine but also shortens the treatment regimen. However, immunological mechanisms behind the perceived benefits of this combination therapy have not been investigated in detail. In the present study, we hypothesized that potential use of drugs that target the host in addition to the parasite might represent an alternative strategy for combination therapy. We investigated immune responses generated in Leishmania donovani infected animals (hamsters and mice) treated with combination of CpG-ODN-2006 and miltefosine at short dose regimen. Infected animals were administered CpG-ODN-2006 (0.4 mg/kg, single dose), as free and liposomal form, either alone or in combination with miltefosine for 5 consecutive days and parasite clearance was evaluated at day 4 and 7 post treatment. Animals that received liposomal CpG-ODN-2006 (lipo-CpG-ODN-2006) and sub-curative miltefosine (5 mg/kg) showed the best inhibition of parasite multiplication (∼97%) which was associated with a biased Th1 immune response in. Moreover, compared to all the other treated groups, we observed increased mRNA expression levels of pro-inflammatory cytokines (IFN-γ, TNF-α and IL-12) and significantly suppressed levels of Th2 cytokines (IL-10 and TGF-ß) on day 4 post treatment in animals that underwent combination therapy with lipo-CpG-ODN-2006 and sub-curative miltefosine. Additionally, same therapy also induced heightened iNOS mRNA levels and NO generation, increased IgG2 antibody level and strong T-cell response in these hamsters compared with all the other treated groups. Collectively, our results suggest that combination of lipo-CpG-ODN-2006 and sub-curative miltefosine generates protective T-cell response in an animal model of visceral leishmaniasis which is characterized by strong Th1 biased immune response thereby underlining our hypothesis that combination therapy, at short dose regimen can be used as a novel way of treating visceral leishmaniasis.