Genomic and Metabolomic Polymorphism among Experimentally Selected Paromomycin-Resistant Leishmania donovani Strains.

Understanding the mechanism(s) underpinning drug resistance could lead to novel treatments to reverse the increased tolerance of a pathogen. In this study, paromomycin (PMM) resistance (PMMr) was induced in three Nepalese clinical strains of Leishmania donovani with different inherent susceptibilities to antimony (Sb) drugs by stepwise exposure of promastigotes to PMM. Exposure to PMM resulted in the production of mixed populations of parasites, even though a single cloned population was used at the start of selection. PMM 50% inhibitory concentration (IC50) values for PMMr parasites varied between 104 and 481 µM at the promastigote stage and 32 and 195 µM at the intracellular amastigote stage. PMM resistance was associated with increased resistance to nitric oxide at the amastigote stage but not the promastigote stage (P < 0.05). This effect was most marked in the Sb-resistant (Sbr) PMMr clone, in which PMM resistance was associated with a significant upregulation of glutathione compared to that in its wild type (P < 0.05), although there was no change in the regulation of trypanothione (detected in its oxidized form). Interestingly, PMMr strains showed an increase in either the keto acid derivative of isoleucine (Sb intermediate PMMr) or the 2-hydroxy acids derived from arginine and tyrosine (Sb susceptible PMMr and Sbr PMMr). These results are consistent with the recent finding that the upregulation of the branched-chain amino acid aminotransferase and d-lactate dehydrogenase is linked to PMMr In addition, we found that PMMr is associated with a significant increase in aneuploidy during PMM selection in all the strains, which could allow the rapid selection of genetic changes that confer a survival advantage.

Enhancing venetoclax activity in acute myeloid leukemia by co-targeting MCL1.

Targeted therapies are frequently combined with standard cytotoxic drugs to enhance clinical response. Targeting the B-cell lymphoma 2 (BCL-2) family of proteins is an attractive option to combat chemoresistance in leukemia. Preclinical and clinical studies indicate modest single-agent activity with selective BCL-2 inhibitors (for example, venetoclax). We show that venetoclax synergizes with cytarabine and idarubicin to increase antileukemic efficacy in a TP53-dependent manner. Although TP53 deficiency impaired sensitivity to combined venetoclax and chemotherapy, higher-dose idarubicin was able to suppress MCL1 and induce cell death independently of TP53. Consistent with an MCL1-specific effect, cell death from high-dose idarubicin was dependent on pro-apoptotic Bak. Combining higher-dose idarubicin with venetoclax was able to partially overcome resistance in Bak-deficient cells. Using inducible vectors and venetoclax to differentially target anti-apoptotic BCL-2 family members, BCL-2 and MCL1 emerged as critical and complementary proteins regulating cell survival in acute myeloid leukemia. Dual targeting of BCL-2 and MCL1, but not either alone, prolonged survival of leukemia-bearing mice. In conclusion, our findings support the further investigation of venetoclax in combination with standard chemotherapy, including intensified doses of idarubicin. Venetoclax should also be investigated in combination with direct inhibitors of MCL1 as a chemotherapy-free approach in the future.

Seroprevalence of hepatitis B virus and hepatitis C virus co-infection among people living with HIV/AIDS visiting antiretroviral therapy centres in Nepal: a first nationally representative study.

OBJECTIVES: To assess the prevalence of human immunodeficiency virus (HIV), hepatitis C virus (HCV), and hepatitis B virus (HBV) co-infections among people living with HIV (PLHIV) in Nepal. METHODS: A sample of 677 PLHIV representing key affected populations (KAP) in Nepal, who were undergoing antiretroviral (ART) therapy in ART clinics around the country, were voluntarily enrolled in the study. Rapid kit-based testing followed by ELISA for validation was performed, focusing on HBV surface antigen (HBsAg) and antibodies against HCV (anti-HCV). A multivariate logistic regression model was used to identify factors associated with HBV and HCV co-infection. RESULTS: HCV and HBV co-infection among the 677 PLHIV was found to be 19% (95% confidence interval (CI) 16.6-22.7%) and 4.4% (95% CI 3.1-6.6%), respectively. The Eastern Region had the highest percentage of HCV infection (48%). The age group with the highest rates of co-infection was 30-39 years (58% and 70%, respectively, for HCV and HBV co-infection). After adjusting for confounding, males were more likely to have HBV co-infection than females (adjusted odds ratio (AOR) 4.61, 95% CI 1.42-14.98). Similarly, PLHIV who were male (AOR 5.7, 95% CI 2.06-15.98), had a secondary level of education (AOR 3.04, 95% CI 1.06-8.70), or who were drug users (AOR 28.7, 95% CI 14.9-55.22) were significantly more likely to have HCV co-infection. CONCLUSION: This first ever national assessment of HIV, HBV, and HCV co-infection performed among PLHIV in Nepal demonstrates that HCV and HBV infections are a health threat to this population and that interventions are required to mitigate the effects of co-infection and to prevent further morbidity and mortality.

Patterns of Orthopaedic Injuries among Motorbike Accident Admissions Presenting to a Tertiary Care Hospital in Kathmandu.

BACKGROUND: The anecdotal burden of morbidity secondary to motorbike accidents is thought to enormous by personnel involved in trauma care. The objective of the present study is to ascertain patterns and association of injuries and causes of mortality in a cohort of motorbike accident victims. METHODS: A retrospective chart review of 1337 motorbike accident admissions B and B Hospital in Kathmandu between January 2009 and December 2010 was undertaken. RESULTS: Majority of victims were between 20 to 50 years [1230 (92%) males]. Lower extremity injuries comprised on 816 (61% of total admissions). Tibia fractures were the most common injury in isolation, multiple injury, as well as polytrauma, and a vast majority of these were open fractures. Multiple injuries were seen in 82 (6.1%) patients and 33 (2.5% of Total) patients were polytraumatized. An amputation was necessary in 16 (1.2%) patients and emergent fasciotomy for compartment syndrome was necessary in 23 (1.7%) cases of tibia fractures and 39 (2.2%) of foot and ankle injuries. A floating joint injury was present in 24 (1.8%) patients.. A fat embolism syndrome (FES) was diagnosed in 8 (0.6%) patients. The overall mortality was 0.45% (6 patients). CONCLUSIONS: Motorbike accident is an important cause of morbidity and mortality in Kathmandu, and often involves a very productive age group. There is an imminent need to address this public health problem.

In vitro selection of miltefosine resistance in promastigotes of Leishmania donovani from Nepal: genomic and metabolomic characterization.

In this study, we followed the genomic, lipidomic and metabolomic changes associated with the selection of miltefosine (MIL) resistance in two clinically derived Leishmania donovani strains with different inherent resistance to antimonial drugs (antimony sensitive strain Sb-S; and antimony resistant Sb-R). MIL-R was easily induced in both strains using the promastigote-stage, but a significant increase in MIL-R in the intracellular amastigote compared to the corresponding wild-type did not occur until promastigotes had adapted to 12.2 µM MIL. A variety of common and strain-specific genetic changes were discovered in MIL-adapted parasites, including deletions at the LdMT transporter gene, single-base mutations and changes in somy. The most obvious lipid changes in MIL-R promastigotes occurred to phosphatidylcholines and lysophosphatidylcholines and results indicate that the Kennedy pathway is involved in MIL resistance. The inherent Sb resistance of the parasite had an impact on the changes that occurred in MIL-R parasites, with more genetic changes occurring in Sb-R compared with Sb-S parasites. Initial interpretation of the changes identified in this study does not support synergies with Sb-R in the mechanisms of MIL resistance, though this requires an enhanced understanding of the parasite's biochemical pathways and how they are genetically regulated to be verified fully.

Lack of correlation between the promastigote back-transformation assay and miltefosine treatment outcome.

OBJECTIVES: Widespread antimony resistance in the Indian subcontinent has enforced a therapy shift in visceral leishmaniasis treatment primarily towards miltefosine and secondarily also towards paromomycin. In vitro selection of miltefosine resistance in Leishmania donovani turned out to be quite challenging. Although no increase in IC50 was detected in the standard intracellular amastigote susceptibility assay, promastigote back-transformation remained positive at high miltefosine concentrations, suggesting a more 'resistant' phenotype. This observation was explored in a large set of Nepalese clinical isolates from miltefosine cure and relapse patients to assess its predictive value for patient treatment outcome. METHODS: The predictive value of the promastigote back-transformation for treatment outcome of a set of Nepalese L. donovani field isolates (n = 17) derived from miltefosine cure and relapse patients was compared with the standard susceptibility assays on promastigotes and intracellular amastigotes. RESULTS: In-depth phenotypic analysis of the clinical isolates revealed no correlation between the different susceptibility assays, nor any clear link to the actual treatment outcome. In addition, the clinical isolates proved to be phenotypically heterogeneous, as reflected by the large variation in drug susceptibility among the established clones. CONCLUSIONS: This in vitro laboratory study shows that miltefosine treatment outcome is not necessarily exclusively linked with the susceptibility profile of pre-treatment isolates, as determined in standard susceptibility assays. The true nature of miltefosine treatment failures still remains ill defined.

Extra and Intra-articular Synovial Chondromatosis.

Synovial chondromatosis is not so rare intra-articular condition secondary to synovial metaplasia, that affects the knee joint. Extra-articular synovial chondromatosis however is an extremely rare condition that usually involves the synovial sheath or bursa of the foot or hand. We present two cases of synovial chondromatosis, one intra and one extra-articular. The first case was a 25 year old lady who presented with pain, swelling and restricted range of motion of left knee and was found to have an intra-articular synovial chondromatosis which was treated successfully by joint debridement. The second case was that of a 22 year old man who presented with right knee pain and was diagnosed to have an extra-articular synovial chondromatosis of his right medial hamstring tendon sheath, excision of which resulted in complete relief of symptoms.

Experimental selection of paromomycin and miltefosine resistance in intracellular amastigotes of Leishmania donovani and L. infantum.

Although widespread resistance of Leishmania donovani and L. infantum against miltefosine (MIL) and paromomycin (PMM) has not yet been demonstrated, both run the risk of resistance selection. Unraveling the dynamics and mechanisms of resistance development is key to preserve drug efficacy in the field. In this study, resistance against PMM and MIL was experimentally selected in vitro in intracellular amastigotes of several strains of both species with different antimony susceptibility background. To monitor amastigote susceptibility, microscopic determination of IC50-values and promastigote back-transformation assays were performed. Both techniques were also used to evaluate the susceptibility of field isolates from MIL-relapse patients. PMM-resistance could readily be selected in all species/strains, although promastigotes remained fully PMM-susceptible. Successful MIL-resistance selection was demonstrated only by promastigote back-transformation at increasing MIL-concentrations upon successive selection cycles. Important to note is that amastigotes with the MIL-resistant phenotype could not be visualized after Giemsa staining; hence, MIL-IC50-values showed no shift. The same phenomenon was observed in a set of recent clinical isolates from MIL-relapse patients. This study clearly endorses the need to use intracellular amastigotes for PMM- and MIL-susceptibility testing. When monitoring MIL-resistance, promastigote back-transformation should be used instead of the standard Giemsa staining. In-depth exploration of the mechanistic background of this finding is warranted.

Epistaxis in visceral leishmaniasis with hematological correlation.

Objective. To study the prevalence of epistaxis in visceral leismaniasis and its correlation with hematological profile. Methods. Out of 80 diagnosed cases of visceral leishmaniasis, 19 patients with epistaxis were included in the study. Diagnosis was made by Rk-39 from peripheral smear and LD bodies from bone marrow. Before starting anti-kala-azar treatment, nasal examination findings and hematological profile were noted. Study Design. Prospective cross-sectional hospital-based study. Results. Epistaxis was found in the age group of 7-66 years. Epistaxis was observed in 19 (23.8%) cases. One patient died because of epistaxis and neck hematoma. Conclusion. Epistaxis is a common ENT finding in endemic area of visceral leishmaniasis like our case.

Increased metacyclogenesis of antimony-resistant Leishmania donovani clinical lines.

Mathematical models predict that the future of epidemics of drug-resistant pathogens depends in part on the competitive fitness of drug-resistant strains. Considering metacyclogenesis (differentiation process essential for infectivity) as a major contributor to the fitness of Leishmania donovani, we tested its relationship with pentavalent antimony (SbV) resistance in clinical lines. Different methods for the assessment of metacyclogenesis were cross-validated: gene expression profiling (META1 and SHERP), morphometry (microscopy and FACS), in vitro infectivity to macrophages and resistance to complement lysis. This was done on a model constituted by 2 pairs of reference strains cloned from a SbV-resistant and -sensitive isolate. We selected the most adequate parameter and extended the analysis of metacyclogenesis diversity to a sample of 20 clinical lines with different in vitro susceptibility to the drug. The capacity of metacyclogenesis, as measured by the complement lysis test, was shown to be significantly higher in SbV-resistant clinical lines of L. donovani than in SbV-sensitive lines. Together with other lines of evidence, it is concluded that L. donovani constitutes a unique example and model of drug-resistant pathogens with traits of increased fitness. These findings raise a fundamental question about the potential risks of selecting more virulent pathogens through massive chemotherapeutic interventions.

Clinical, MRI and arthroscopic correlation in internal derangement of knee.

BACKGROUND: The traumatic or degenerative internal derangement of the knee requires certain investigations for the establishment of diagnosis, in addition to clinical history and a thorough physical examination. The use of arthrography and arthroscopy improves the accuracy of the diagnosis. MRI scanning of the knee joint has often been regarded as the noninvasive alternative to diagnostic arthroscopy. OBJECTIVE: The purpose of the study was to correlate clinical and low field MRI findings with arthroscopy in internal derangement of the knee. METHODS: Forty one patients with suspected internal derangement of the knee were subjected to MR examination followed by arthroscopy. Clinical criteria used were history, mode of injury, Mc Murray, Apley grinding, Thessaly test for meniscal injury. Drawer test was considered to be essential for clinical diagnosis of cruciate ligament injury. MRI of the knee was performed in low field open magnet (0.35T, Magnetom C, Seimens). Arthroscopy was done within two months of MR examination and was considered gold standard for the internal derangement of the knee. RESULTS: The sensitivity, specificity, diagnostic accuracy of clinical examination were 96.1%, 33.3% and 73.1% respectively for medial meniscal tear; 38.4%, 96.4% and 78.1% respectively for lateral meniscal tear. The sensitivity, specificity, diagnostic accuracy of MRI were 92.3%,100% and 95.1% for medial meniscal tear; 84.6%96.4% and 92.6% respectively for lateral meniscal tear. CONCLUSION: Clinical examination showed higher sensitivity for medial meniscal tear compared to MRI, however with low specificity and diagnostic accuracy. Low field MRI showed high sensitivity, specificity, diagnostic accuracy for meniscal and cruciate ligament injury, in addition to associated derangement like articular cartilage damage, synovial thickening.

Will visceral leishmaniasis be eliminated from Nepal? A review of recent (1994-2006) control efforts.

The Visceral Leishmaniasis (VL) control program in Nepal launched in 1993 includes provision of free diagnostic test, treatment along with vector control by indoor residual spraying. However, even after 14 years the disease is far from being controlled. Elimination of VL by 2015 has recently been identified as a regional priority with high level of political commitment. We analyzed the VL control effort in Nepal over the period 1994-2006 and tried to formulate recommendations for the VL elimination initiative. To document performance of the VL control program in Nepal we used literature review and a case study. First, we reviewed articles on VL in Nepal published in medical journals through Pubmed, ISI Web of Science, Google scholar and by hand searching. Secondly, the grey literature, mainly the reports on VL drafted by the Ministry of Health was reviewed for the period 1994-2006. Thirdly, a case study is presented to analyze the performance of the VL elimination program in a "pilot district", where the program was launched in 2006. There are only few studies available on VL in Nepal discussing the epidemiology, diagnosis, treatment, vector bionomics, human behavior and prevention. The review of the grey literature from 1994-2006 revealed that the VL incidence rate remained almost constant since 1993 despite the control efforts. The case study showed that there is a lack of trained human resources, laboratory facilities and treatment guidelines which is hindering the decentralization of the VL elimination program.

Development and evaluation of different PCR-based typing methods for discrimination of Leishmania donovani isolates from Nepal.

INTRODUCTION: Leishmania donovani, the causative agent of visceral leishmaniasis in the Indian subcontinent, has been reported to be genetically homogeneous. In order to support ongoing initiatives to eliminate the disease, highly discriminative tools are required for documenting the parasite population and dynamics. METHODS: Thirty-four clinical isolates of L. donovani from Nepal were analysed on the basis of size and restriction endonuclease polymorphisms of PCR amplicons from kinetoplast minicircle DNA, 5 nuclear microsatellites, and nuclear loci encoding glycoprotein 63, cysteine proteinase B, and hydrophilic acylated surface protein B. We present and validate a procedure allowing standardized analysis of kDNA fingerprint patterns. RESULTS: Our results show that parasites are best discriminated on the basis of kinetoplast minicircle DNA (14 genotypes) and 1 microsatellite defining 7 genotypes, while the remaining markers discriminated 2 groups or were monomorphic. Combination of all nuclear markers revealed 8 genotypes, while extension with kDNA data yielded 18 genotypes. CONCLUSION: We present tools that allow discrimination of closely related L. donovani strains circulating in the Terai region of Nepal. These can be used to study the micro-epidemiology of parasite populations, determine the geographical origin of infections, distinguish relapses from re-infection, and monitor the spread of particular variants.

Clinical risk factors for therapeutic failure in kala-azar patients treated with pentavalent antimonials in Nepal.

Drug-related factors and parasite resistance have been implicated in the failure of pentavalent antimonials (Sb(v)) in the Indian subcontinent; however, little information is available on host-related factors. Parasitologically confirmed kala-azar patients, treatment naïve to Sb(v), were prospectively recruited at a referral hospital in Nepal and were treated under supervision with 30 doses of quality-assured sodium stibogluconate (SSG) 20mg/kg/day and followed for 12 months to assess cure. Analysis of risk factors for treatment failure was assessed in those receiving >or=25 doses and completing 12 months of follow-up. One hundred and ninety-eight cases were treated with SSG and the overall cure rate was 77.3% (153/198). Of the 181 cases who received >or=25 doses, 12-month follow-up data were obtained in 169, comprising 153 patients (90.5%) with definite cure and 16 (9.5%) treatment failures. In the final logistic regression model, increased failure to SSG was significantly associated with fever for >or=12 weeks [odds ratio (OR)=7.4], living in districts bordering the high SSG resistance zone in Bihar (OR=6.1), interruption of treatment (OR=4.3) and ambulatory treatment (OR=10.2). Early diagnosis and supervised treatment is of paramount importance to prevent treatment failures within the control programme.

Determinants of bednet ownership and use in visceral leishmaniasis-endemic areas of the Indian subcontinent.

OBJECTIVE: To document ownership and use of bednets with its determinants in the visceral leishmaniasis (VL)-endemic region where mainly non-insecticide impregnated nets are available through commercial channels, and bednets are being considered as a leishmaniasis vector control measure. METHODS: In August-September 2006, semi-structured household (HH) questionnaires and observation guides were used in a random sample of 1330 HHs in VL-endemic districts of India and Nepal to collect data on VL knowledge, HH socio-economic status, bednet ownership and use patterns. An asset index was constructed to allow wealth ranking of the HH. A binary logistic response General Estimating Equations model was fitted to evaluate the determinants of bednet ownership and use. RESULTS: The proportion of HHs with at least one bednet purchased on the commercial market was 81.5% in India and 70.2% in Nepal. The bednets were used in all seasons by 50.6% and 54.1% of the Indian and Nepalese HH owning a bed net. There was striking inequity in bednet ownership: only 38.3% of the poorest quintile in Nepal owned at least one net, compared to 89.7% of the wealthiest quintile. In India, the same trend was observed though somewhat less pronounced (73.6%vs. 93.7%). Multivariate analysis showed that poverty was an important independent predictor for not having a bednet in the HH [OR 5.39 (2.90-10.03)]. CONCLUSION: Given the inequity in commercial bednet ownership, free distribution of insecticide-treated bednets to the general population seems imperative to achieve a mass effect on vector density.

Chronic cough in thymoma.

Cough is one of the most common symptom for which patients seek medical attention from primary care physicians and pulmonologists. Although tuberculosis and other lung infections are common throughout the developing world, they are not among the most common causes of chronic cough. We report a case of a 23 years old male who presenting to the outpatient clinic with chronic cough not responding to regular and symptomatic treatment that was diagnosed to have thymoma. After all the common causes for chronic cough have been ruled out, unusual causes should also be considered in the differential diagnosis.

Comparative study of kala-azar vector control measures in eastern Nepal.

This study was conducted to explore the most effective vector control tool among indoor residual spraying (IRS), long lasting insecticidal nets (LLINs) and ecological vector management (EVM) as a part of the regional visceral leishmaniasis elimination initiative. Alpha-cypermethrin as IRS, PermaNet as LLINs and plastering the inner walls of houses with lime as EVM were the interventions. One baseline and three follow-up entomological surveys were carried out in all arms using CDC miniature light traps (LT) and mouth aspirators. Comparisons were made between intervention arms and control arms with pre-intervention and post-intervention vector densities. Light traps were found more efficient in the collection of Phlebotomus argentipes in comparison with aspiration. Vector densities were significantly low in both IRS arm (p=0.009 in LT and p<0.001 in aspirator collections) and LLIN arm (p=0.019 in LT and p=0.023 in aspirator collections) in comparison with control arm. However, in EVM arm, there was no significant difference in P. argentipes sand fly density in comparison with control arm (p=0.785) in LT collections in follow-up surveys. Hence, IRS was found most effective control measure to decrease vector density. LLINs were also found effective and can be considered as a promising alternative vector control tool in VL elimination initiative.

Visceral leishmaniasis on the Indian sub-continent: a multi-centre study of the costs of three interventions for the control of the sandfly vector, Phlebotomus argentipes.

The sandflies that transmit the parasites causing human visceral leishmaniasis (VL) can be controlled by several methods, including indoor residual spraying (IRS), the use of long-lasting insecticidal nets (LLIN) and ecological vector management (EVM). The financial costs of each of these three methods of sandfly control have recently been assessed and compared, in a multi-centre study based on the Indian sub-continent. In each of the four study sites (two in Nepal and one each in India and Bangladesh), 24 neighbourhoods were randomly selected in districts with high incidences of VL. The costs of the three interventions were then prospectively assessed in each study neighbourhood, in the local currency, and then converted to U.S. dollars at the prevailing exchange rate in the country concerned. The costs of IRS, which ranged from U.S.$2.4-11.7 (mean = U.S.$5.9) per household-year, were greater than those of LLIN (U.S.$3.5-5.1/household-year, with a mean of U. S.$4.5) but less than those of EVM (U. S.$5.0- 14.0/household-year, with a mean of U.S.$8.7). These results indicate that LLIN and IRS may be the cheaper options for the control of sandflies on the Indian sub-continent, and that EVM should perhaps only be taken up as a complimentary and voluntary method. Various combinations of these interventions (based on country-specific social and economic factors) may, however, be the best and most cost-effective choice.

Complete inhibition of spontaneous activity in neuronal networks in vitro by deltamethrin and permethrin.

Types I and II pyrethroid insecticides cause temporally distinct decreases in voltage-gated sodium channel (VGSC) inactivation rates that are proposed to underlie their characteristic differences in toxicity signs. How alterations in VGSC channel function give rise to the characteristic differences in signs of pyrethroid intoxication is not completely understood, particularly those changes that occur in functional networks of interconnected neurons. To characterize better pyrethroid actions at the network level, effects of the Type I pyrethroid permethrin (PM) and the Type II pyrethroid deltamethrin (DM) on spontaneous glutamate network-dependent spikes and bursts were investigated in primary cultures of frontal cortex or spinal cord neurons grown on microelectrode arrays (MEAs). Fast GABAergic transmission was blocked by BIC, and concentration-dependent effects of DM (1nM to 5microM) and PM (10nM to 50microM) were examined. Both compounds caused concentration-dependent reductions in the network spike and burst rates. DM was more potent than PM, with IC(50) values of approximately 0.13 and approximately 4microM for inhibition of spike rate in cortical and spinal cord neurons, respectively. Both compounds decreased the percentage of spikes that occurred within a burst and increased the interspike interval within bursts. Onset of effects was rapid, but recovery from total activity loss was not readily achievable. Individual neurons responded heterogeneously; activity of most declined monophasically, but activity in others exhibited biphasic responses with increases followed by decreases in activity. In spinal cord, DM caused a greater number of biphasic responses (29%) than PM (10%). These results demonstrate that both DM and PM inhibit activity of glutamatergic networks, but with different potencies.

Erythrocyte glutathione status in human visceral leishmaniasis.

A glutathione redox cycle is a major antioxidant defense system for the detoxification of reactive oxygen species within erythrocytes. Reactive oxygen species such as superoxide anions, hydrogen peroxide and hydroxyl radicals are generated as a host defense mechanism for killing of engulfed Leishmania donovani, a causative agent of visceral leishmaniasis, are capable of damaging lipids and other biomolecules when produced in excess. Erythrocytes are most vulnerable to Reactive oxygen species. In present study we aimed to evaluate erythrocyte reduced glutathione (GSH) levels as an antioxidant and erythrocyte malondialdehyde (MDA) as a marker of lipid peroxidation. The study included twenty-five Visceral leishmaniasis patients and they were followed up after their complete chemotherapy with antileishmanial drugs (sodium stibogluconate) for 30 days. Forty six age and sex matched healthy individuals were taken as controls. GSH levels in erythrocytes of visceral leishmaniasis patients were increased in spite of significant increased erythrocyte MDA as compared to controls. Whereas erythrocyte GSH and MDA levels of follow up patients were decreased as compared to patients before treatment groups. We concluded that visceral leishmaniasis patients are in oxidative stress which most likely induces the endogenous antioxidant such as GSH or its poor utilization by cells.