Isolation and propagation of classical swine fever virus in porcine Wharton's Jelly mesenchymal stem cells.

Classical Swine Fever (CSF) is an extremely infectious and deadly disease of pigs and wild boars caused by the CSF virus (CSFV) which is a member of the Pestivirus genus and the family Flaviviridae. This study was designed to detect the permissibility and replication of CSFV in mesenchymal stem cells (MSCs) monolayer derived from Porcine Wharton's jelly. Porcine Wharton's jelly MSCs (pWJ-MSCs) were ex vivo expanded and propagated for more than 81 generations and third passage pWJ-MSCs were characterized as per standard criteria i.e., growth characteristics, trilineage differentiation potential and molecular characterization for pluripotency and stem cell surface markers. Porcine WJ tissue samples found negative for CSFV by RT-PCR test were processed further for the isolation of pWJ-MSCs and CSFV was propagated over the characterized pWJ-MSCs monolayer. No cytopathic effect was observed, which was consistent with non-cytopathic nature of CSFV. The replication of CSFV in pWJ-MSCs was affirmed by RT-PCR and demonstration of viral antigen in the cytoplasm of virus infected cells by immuno-staining technique. In total, three different CSFV isolates were propagated in pWJ-MSCs. Primary pWJ-MSCs permitted CSFV replication to good titer. To the best of our information, this is the first ever report of isolation of CSFV in pWJ-MSCs.

Progeria: A Rare Genetic Syndrome.

An uncommon deadly genetic situation symbolized by the presence of rapid maturation in infants is called as the Hutchinson-Gilford Progeria Syndrome. The term basically is meant as 'prematurely old' taken from the Greek meanings. The selective cause behind this syndrome is usually a mutation in a gene called LMNA. The product of this LMNA gene which is a protein i.e. Lamin-A is considered to be responsible for anatomical framing which clasps the nuclei of the cell, well organized and together. But, the recent investigations prove a deformity in the protein i.e. Lamin-A that leads to the non-stability of the nuclei an thus gives rise to the deadly situation of untimely ageing in the children popularly known as Progeria. The literature review investigation provided pivotal information about the therapeutic researches related to the syndrome, the mutational causes and the basic information including the major and minor symptoms generally shown by the patients affected with Hutchinson-Gilford Progeria Syndrome. Investigations on this rare, uncommon disease i.e. Progeria had begun a couple of years back and in some of the researches many important aspects about the causes and possible curative drugs related to the disease which can help the patients in leading a normal life with lesser side effects and symptoms have also been discussed. Further studies will more clearly clarify the possible curative agents and unrevealed mechanisms of the disease which will help the scientists to develop measures which can provide more beneficial and healthy life to the patients with lesser complications.

Design and synthesis of imidazolidinone derivatives as potent anti-leishmanial agents by bioisosterism.

Bioisosterism is a useful strategy in rational drug design to improve pharmacodynamic and pharmacokinetic properties of lead compounds. Imidazolidinones have been reported as potent kinase inhibitors and antileishmanial agents. In this study, bioisosteres of imidazolidinones (compounds 1-3) were evaluated for their antileishmanial properties. The modified imidazolidinones exhibited potent antileishmanial activity against extracellular as well as intracellular Leishmania donovani parasites in nanomolar concentrations. The selectivity index of these compounds on host cells was found to be more than 1000, emphasizing their specificity toward the parasite. Using SwissTargetPrediction software, we assessed the potential targets of these compounds and found MAPK as the most probable target. To in vitro validate, we developed a novel in vitro kinase assay that mimics the in vivo nature of the functional kinome. Compounds 1-3 displayed specific inhibition of parasite kinase activity accompanied by an increase in intracellular sodium levels in the parasites. This might be the effect of kinase inhibition that regulates sodium homeostasis through Na-ATPases. Finally, the compound-treated parasites underwent apoptosis-like death. This study represents bioisoterism as a novel approach for drug design to establish the structure-activity relationship, which in turn helps to improve the therapeutic activity of lead compounds.

Genetic variations of IL-10: Identification of novel variations and evaluation of the impact of the SNPs/haplotype in the promoter region with the progression of Oral Squamous Cell Carcinoma in Indian population.

The correlation of interleukin 10 (IL-10) with the outbreak and progression of cancer has been well established as it contributes to tumor immune evasion. Convincing number of evidences has been accumulated to reflect the critical correlation between IL-10 polymorphism and tumorogenesis. Several polymorphic sites at promoter regions have been reported to be associated with cancer susceptibility. The purpose of this study was to examine the effect of modulated genotypes in the promoter region of IL-10 gene with life-style habits in oral squamous cell carcinoma (OSCC) in the Indian population. A total of 300 subjects (100 OSCC, 50 precancer and 150 healthy controls) were recruited in this study. The IL-10 promoter region was amplified in 14 overlapping fragments by PCR and further screened through the high throughput technique of denaturing high-performance liquid chromatography (dHPLC) followed by sequencing. We identified three novel variations at positions (-924, -1045 & -1066); we also found some known SNPs (-592C/A, -657G/A, -851G/A, -819C/T, -1082A/G). The identified novel variations were submitted to the NCBI Gene Bank (accession numbers KT153594, KT291742 and KT291743). We also noticed a significant association of polymorphisms (-592C/A, -819C/T and -1082A/G) individually as well as in combination (haplotypes) along with lifestyle habits for the risk of oral carcinoma (p<0.0001). We have reported three novel SNPs in the Indian population for the first time, and these SNPs may be associated with OSCC. Besides, we showed the first evidence of IL-10 haplotypes, i.e., CCG and CTG, may act as a biomarker for early detection of oral pre-cancerous/cancerous lesions or treatment management of oral carcinoma.

A role for adaptor protein complex 1 in protein targeting to rhoptry organelles in Plasmodium falciparum.

The human malaria parasite Plasmodium falciparum possesses sophisticated systems of protein secretion to modulate host cell invasion and remodeling. In the present study, we provide insights into the function of the AP-1 complex in P. falciparum. We utilized GFP fusion constructs for live cell imaging, as well as fixed parasites in immunofluorescence analysis, to study adaptor protein mu1 (Pfµ1) mediated protein trafficking in P. falciparum. In trophozoites Pfµ1 showed similar dynamic localization to that of several Golgi/ER markers, indicating Golgi/ER localization. Treatment of transgenic parasites with Brefeldin A altered the localization of Golgi-associated Pfµ1, supporting the localization studies. Co-localization studies showed considerable overlap of Pfµ1 with the resident rhoptry proteins, rhoptry associated protein 1 (RAP1) and Cytoadherence linked asexual gene 3.1 (Clag3.1) in schizont stage. Immunoprecipitation experiments with Pfµ1 and PfRAP1 revealed an interaction, which may be mediated through an intermediate transmembrane cargo receptor. A specific role for Pfµ1 in trafficking was suggested by treatment with AlF4, which resulted in a shift to a predominantly ER-associated compartment and consequent decrease in co-localization with the Golgi marker GRASP. Together, these results suggest a role for the AP-1 complex in rhoptry protein trafficking in P. falciparum.

Vitamin B12 Grafted Layer-by-Layer Liposomes Bearing HBsAg Facilitate Oral Immunization: Effect of Modulated Biomechanical Properties.

Adhesion forces of nanoparticulate materials toward biological membrane are crucial for designing a delivery system for therapeutic molecules and vaccines. The present study aims to investigate the impact of surface roughness of the nanoparticulate system in oral delivery of antigen and its targeting to toward intestinal antigen presenting cells. To evaluate this hypothesis, layer-by-layer coated liposomes (LBL-Lipo) were fabricated using sodium alginate and Vitamin B12 conjugated Chitosan (VitB12-Chi) as anionic and cationic polyelectrolyte, respectively. Change in surface roughness was observed on changes in pH from gastric to intestinal conditions attributed to increase and decrease in charge density on VitB12-Chi. Surface roughness was measured in terms of root-mean-square measured by topographical analysis using atomic force microscopy. LBL-Lipo were further characterized for their size, zeta potential, and release behavior to evaluate the potential for oral vaccine delivery. In vitro cell uptake in macrophage cells (J-744) shows about 2- and 3.1-fold increased uptake of rough LBL-Lipo over smooth LBL-Lipo at 37 °C (endocytosis) and 4 °C (endocytosis inhibition) indicating improved biological interaction. Further in vivo immunization study revealed that prototype formulations were able to produce 4.8- and 3.3-fold higher IgG and IgA levels in serum and feces, respectively, in comparison to smooth LBL-Lipo.

The dilemma of arranged marriages in people with epilepsy. An expert group appraisal.

INTRODUCTION: Matrimony remains a challenging psychosocial problem confronting people with epilepsy (PWE). People with epilepsy are less likely to marry; however, their marital prospects are most seriously compromised in arranged marriages. AIMS: The aim of this study was to document marital prospects and outcomes in PWE going through arranged marriage and to propose optimal practices for counseling PWE contemplating arranged marriage. METHODS: A MEDLINE search and literature review were conducted, followed by a cross-disciplinary meeting of experts to generate consensus. RESULTS: People with epilepsy experience high levels of felt and enacted stigma in arranged marriages, but the repercussions are heavily biased against women. Hiding epilepsy is common during marital negotiations but may be associated with poor medication adherence, reduced physician visits, and poor marital outcome. Although divorce rates are generally insubstantial in PWE, divorce rates appear to be higher in PWE undergoing arranged marriages. In these marriages, hiding epilepsy during marital negotiations is a risk factor for divorce. CONCLUSIONS: In communities in which arranged marriages are common, physicians caring for PWE are best-equipped to counsel them about their marital prospects. Marital plans and aspirations should be discussed with the family of the person with epilepsy in a timely and proactive manner. The benefits of disclosing epilepsy during marital negotiations should be underscored.

Lymph node abscess and cardiac involvement in a patient with nodular lepromatous leprosy (LL) with erythema nodosum leprosum (ENL): a rare occurrence.

With the world's focus on reducing the leprosy patient load to the extent of elimination, finding and reporting the rarer presentations of leprosy becomes important for prompt treatment. Also, these untreated patients may serve as a potential source of infection in community. We report a 35-year old man diagnosed to have lepromatous leprosy and erythema nodosum leprosum with inguinal lymph node abscess and suspected cardiac involvement that proved fatal. We stress the importance of detailed workup to look for associated systemic involvement for timely intervention and favourable outcome.

Potential Chemoprevention of 7,12-Dimethylbenz[a]anthracene Induced Renal Carcinogenesis by Moringa oleifera Pods and Its Isolated Saponin.

Present investigation shows that hydroethanolic extract of Moringa oleifera (MOHE) and its isolated saponin (SM) attenuates DMBA induced renal carcinogenesis in mice. Isolation of SM was achieved by TLC and HPLC and characterization was done using IR and (1)H NMR. Animals were pre-treated with MOHE (200 and 400 mg/kg body weight; p.o), BHA as a standard (0.5 and 1 %) and SM (50 mg/kg body weight) for 21 days prior to the administration of single dose of DMBA (15 mg/kg body weight). Administration of DMBA significantly (p < 0.001) enhanced level of xenobiotic enzymes. It enhanced renal malondialdehyde, with reduction in renal glutathione content, antioxidant enzymes and glutathione-S-transferase. The status of renal aspartate transaminase, alanine transaminase, alkaline phosphatase and total protein content were also found to be decreased along with increase in total cholesterol in DMBA administered mice. Pretreatment with MOHE and SM significantly reversed the DMBA induced alterations in the tissue and effectively suppressed renal oxidative stress and toxicity.

Ameliorative Effects of Operculina turpethum and its Isolated Stigma-5,22dien-3-o-ß-D-glucopyranoside on the Hematological Parameters of Male Mice Exposed to N-Nitrosodimethylamine, a Potent Carcinogen.

OBJECTIVES: Enormous propensity of plants to synthesize a variety of structurally diverse bioactive compounds, has made the plant kingdom a potential source of chemical constituents with various therapeutic values, including antitumor and cytotoxic activities. Blood is a good indicator to determine the physiological and pathological status of man and animal. The objective of the present study is to determine the effect of Operculina turpethum root extract and its isolated glycoside treatment on the hematological parameters in the mice with N-Nitrosodimethylamine (NDMA) induced cancer. MATERIALS AND METHODS: The body weights of the animals were recorded before and after the experiment. Non-coagulated blood was tested for total erythrocyte count, total leukocyte count, hemoglobin, differential leukocyte count (DLC) and for other blood indices. RESULTS: A significant (P < 0.01), (P < 0.001) recovery of the red blood cell and white blood cell counts, packed cell volume and hemoglobin content in the host after 21 day treatment was shown. CONCLUSION: These results show that the extract of Operculina turpethum is relatively safe following oral administration and have possible stimulatory effect on red blood cell production and there was dose dependent therapeutic effect.

Protective Assessment of Euphorbia neriifolia and its Isolated Flavonoid Against N-nitrosodiethylamine-induced Hepatic Carcinogenesis in Male Mice: A Histopathological Analysis.

AIMS: The aim of this study was to examine the impacts of N-nitrosodiethylamine (DENA), a potent environment carcinogen on liver tissue of mice which was attenuated by isolated flavonoid and hydro-ethanolic extract of Euphorbia neriifolia (HEEN) leaves. MATERIALS AND METHODS: Carcinogenicity was induced in albino mice by a single oral administration of DENA (50 mg/kg body weight). The HEEN (150 and 400 mg/kg body weight), butylated hydroxyanisole (BHA; 0.5 and 1%) and E. neriifolia flavonoid (ENF; 50 mg/kg body weight) were estimated to examine the possible anti-cancer potential. RESULTS: DENA exposed animals showed alterations in normal hepatic histo-architecture, which comprised of necrosis (N), dilated sinusoids and vacuolization of the cells. Mice treated with E. neriifolia lower (ENL) and higher (ENH) dose and ENF before intoxicated with DENA showed that the liver cells were normal, with very little necrosis (Day 31). On the other hand, BHA higher (BHAH) and lower (BHAL) dose failed to diminish the abnormalities caused by the DENA. CONCLUSION: Results of the present study suggests that the ENH and ENF protects the hepatic tissue against DENA-induced hepatic carcinoma. The results could also be expressed in the order of ENH> ENF> ENL> BHAH> BHAL.

Anti-carcinogenic potential of Euphorbia neriifolia leaves and isolated flavonoid against N-nitrosodiethylamine-induced renal carcinogenesis in mice.

Anti-carcinogenic potential of hydro-ethanolic extract of Euphorbia neriifolia (EN) leaves and an isolated flavonoid (ENF) was investigated against N-Nitrosodiethylamine (DENA)-induced renal carcinogenesis in mice. Experimental mice were pretreated with 150 and 400 mg/kg body wt of EN, 0.5% and 1% mg/kg body wt of butylated hydroxylanisole (BHA) as a standard antioxidant and 50 mg/kg body wt of ENF for 21 days prior to the administration of a single dose of 50 mg/kg body wt of DENA. Levels of renal markers (urea and creatinine), xenobiotic metabolic enzymes (Cyt P450 and Cyt b5), lipid peroxidation (LPO), antioxidants (SOD, CAT, GST and GSH) and other biochemical parameters--AST, ALT, ALP, total protein (TP), and total cholesterol (TC) were measured to determine the renal carcinogenesis caused by DENA. DENA administration significantly (p < 0.001) decreased the body weight and increased the tissue weight. It significantly (p < 0.001) enhanced the levels of Cyt P450, Cyt b5 and LPO and decreased the levels of SOD, CAT, GST and GSH content. The activities of AST, ALT and ALP and the TP content and renal markers were also significantly decreased (p < 0.001), while TC level was markedly increased after DENA administration, as compared with the normal control group (p < 0.001). Pretreatment with EN and ENF counteracted DENA-induced oxidative stress (LPO) and exerted its protective effects by restoring the levels of antioxidants (SOD, CAT, GST and GSH), biochemical parameters (AST, ALT, ALP, TP and TC), renal markers (urea and creatinine) and xenobiotic enzymes (Cyt P450 and Cyt b5) in renal tissue. In conclusion, the present study showed significant anti-carcinogenic potential of the hydro-ethanolic extract of E. neriifolia and ENF against DENA-induced renal carcinogenicity.

Usage of Conventional PCR Technology for the Detection of HLA-B27 Allele: A Significant Molecular Marker of Ankylosing Spondylitis.

Ankylosing spondylitis is a chronic inflammatory disease that has been linked to the human leukocyte antigen class I allele HLA-B27. More than 90 % of patients with ankylosing spondylitis possess the HLA-B27 allele, but only 1 % of people with HLA-B27 develop the disease. Ankylosing spondylitis predominately affects young males. The present study was planned to find out the involvement of HLA-B27 specific allele in relation to age and sex in symptomatic suspected patients of ankylosing spondylitis using conventional PCR technology. Forty symptomatic patients of ankylosing spondylitis were included in the present study. SSP-PCR technique was used to detect the HLA-B27 specific allele. This study showed (out of 40 symptomatic suspected cases of ankylosing spondylitis only 12 patients were detected positive with HLA-B27 allele, while remaining 28 were negative) that the positivity rate for ankylosing spondylitis with HLA-B27 allele is low. Furthermore, it was observed that the males above 50 years are more prone to develop AS with HLA-B27 specific allele. It could be concluded that the conventional PCR technology is a rapid, sensitive, and confirmatory method for the diagnosis of ankylosing spondylitis.

Diagnostic value of PCR in genitourinary tuberculosis.

Genitourinary tuberculosis is a disease of the genitourinary system which includes the entire urinary tract and reproductive system. Genital tuberculosis is an important cause of female infertility, especially in developing nations like India. In the present study, a total of 257 clinical specimens comprising of endometrial biopsy (109), endometrial curetting (42), menstrual blood (8), semen (17), placenta (11) and urine (70) were collected from patients and subjected for PCR, Culture and AFB detection. The endometrial biopsy, endometrial curetting, menstrual blood, semen, placenta, urine showed 30.2, 45.2,12.5, 5.8, 27.2, 31.4 %, positivity rate for tuberculosis by PCR, 7.3, 9.5, 25.0, 0, 9, 8.5 % by culture and 1.8, 2.3, 0, 0, 0, 2.8 % respectively by AFB smear. Being a novel, rapid technique, PCR is the method of choice for rapid diagnosis and management of genitourinary tuberculosis shared with the other concerned tests. This study reveals that genital tuberculosis can occur in any age group, however, the majority of patients were from reproductive age (nearly 75 % of them were from 20-45 years of age) group.

Lead induced alterations in hemoglobin content during gestation and lactation in Swiss albino mice.

Number of studies in animal models has shown changes in hemoglobin content after lead administration during gestation and lactation, still lead induced hematological changes are not well established. In the present study, hemoglobin content of normal and lead exposed pregnant and lactating Swiss mice were compared. Pregnant females were exposed to heavy metal lead orally on diet containing 4.5% lead nitrate and lead acetate trihydrate during gestation to 3rd week of lactation. Hemoglobin content and blood cell counts were examined on 15th day of gestation and on 1st, 11th and 21st day after birth. The results indicated that in lead intoxicated pregnant females, hemoglobin content decreased. From the results of above study it can be concluded that high levels of lead exposure during gestation and lactation can severely damage heme synthesis.

Chemopreventive Role of Euphorbia neriifolia (Linn) and its Isolated Flavonoid Against N-Nitrosodiethylamine-induced Renal Histopathological Damage in Male Mice.

AIMS: This study is an attempt to evaluate the tissue protective efficacy of isolated flavonoid and hydro-ethanolic extract of Euphorbia neriifolia (HEEN) leaves against N-nitrosodiethylamine (DENA) induced renal carcinoma. MATERIALS AND METHODS: Carcinogenicity was induced in Albino mice by oral administration of DENA (50 mg/kg body weight). The HEEN (150 and 400 mg/kg body weight), Butylated hydroxyanisole (BHA; 0.5 and 1%), and Euphorbia neriifolia flavonoid (ENF; 50 mg/kg body weight) were evaluated for their possible tissue carcinogenesis protective potential. RESULTS: DENA treated animals showed alterations in normal renal histo-architecture, which comprised of necrosis (N) and vacuolization of the cells. On the other hand, the mice treated with Euphorbia neriifolia lower (ENL) and higher (ENH) dose and ENF before intoxicated with DENA showed that the renal cells were normal (Day 31). Whereas, BHA higher (BHAH) and lower (BHAL) dose failed to diminish the abnormalities caused by DENA. CONCLUSIONS: The findings of the present study Suggested that ENH and ENF showed highest renal-protective activity among all the pretreatments. The results could also be expressed in the order of ENH > ENF > ENL > BHAH > BHAL.

Assessment of computer-related health problems among post-graduate nursing students.

The study was conducted to assess computer-related health problems among post-graduate nursing students and to develop a Self Instructional Module for prevention of computer-related health problems in a selected university situated in Delhi. A descriptive survey with co-relational design was adopted. A total of 97 samples were selected from different faculties of Jamia Hamdard by multi stage sampling with systematic random sampling technique. Among post-graduate students, majority of sample subjects had average compliance with computer-related ergonomics principles. As regards computer related health problems, majority of post graduate students had moderate computer-related health problems, Self Instructional Module developed for prevention of computer-related health problems was found to be acceptable by the post-graduate students.

Effect of information education and communication (IEC) programme on knowledge of pregnant mothers regarding prevention and management of warning signs during pregnancy.

The present study was conducted to evaluate the effectiveness of an Information, Education and Communication (IEC) programme on knowledge of pregnant mothers regarding prevention and management of warning signs during pregnancy in a selected health care setting at New Delhi. An evaluative research approach with one group pre-test and post-test design was adopted for the present study. A structured interview schedule was prepared. Purposive non-probability sampling technique was employed to interview 30 pregnant mothers who attended antenatal clinic. Data gathered was analysed and interpreted using both descriptive and inferential statistics. The study revealed that there was maximum knowledge deficit regarding warning signs of pregnancy. IEC programme was effective in enhancing the knowledge of pregnant mothers on prevention and management of warning signs during pregnancy.

Allium sativum Essential Oil Supplementation Reverses the Hepatic Inflammation, Genotoxicity and Apoptotic Effects in Swiss Albino Mice Intoxicated with the Lead Nitrate.

Prolonged lead (Pb) exposure impairs human health due to its interference with physiological and biochemical processes. Therefore, it is necessary to investigate natural therapeutics to alleviate Pb-induced intoxication. In the current investigation, essential oil extracted from the fresh bulbs of Allium sativum was considered as a natural remedy. Initially, in vitro antioxidant and anti-inflammatory activity of A. sativum essential oil (ASEO) were explored. The results reported that ASEO exhibits potent antioxidant and anti-inflammatory potential. Additionally, an in vivo study was conducted to elucidate its preventive role against Lead-nitrate (LN)-induced hepatic damage in Swiss albino mice. The experimental mice were allocated into six groups: Control, LN-intoxicated group (50 mg/kg), LN + ASEO (50 mg/kg), LN + ASEO (80 mg/kg), LN + Silymarin (25 mg/kg), and LN + vehicle oil control group. The entire duration of the study was of 30 days. From the results, it was determined that LN exposure elevated the Pb content in hepatic tissues which subsequently increased the serum biomarkers, inflammatory cytokines (NF-kB, TNF-α, IL-6) as well as apoptotic factors (caspase-3, BAX), all of which contribute to DNA damage. Meanwhile, it reduced anti-inflammatory (IFN-γ and IL-10) and anti-apoptotic factors (Bcl-2). Furthermore, Pb accumulation in hepatic tissues changed the histological architecture, which was linked to necrosis, central vein dilation, inflammatory cell infiltration and Kupffer cell activation. In contrast to this, ASEO administration decreased the Pb content, which in turn reduced the level of serum biomarkers, inflammatory and apoptotic factors. At the same time, it increased the anti-inflammatory and anti-apoptotic factors, thereby reduced DNA damage and restored the hepatic histology. In conclusion, exhaustive research is of the utmost demand to elucidate the precise defense mechanisms of ASEO against LN-induced hepatotoxicity.

Characterization of thiamine pyrophosphokinase of vitamin B1 biosynthetic pathway as a drug target of Leishmania donovani.

Vitamin B1 is an essential cofactor for enzymes involved in the metabolism of carbohydrates, particularly Transketolases. These enzymes are amenable to therapeutic interventions because of their specificity. In the final step of the Vitamin B1 biosynthesis pathway, Thiamine Pyrophosphokinase (TPK) converts thiamin into its active form, Thiamin Pyrophosphate (TPP), allowing researchers to investigate the functional importance of this enzyme and the pathway's dispensability in Leishmania donovani, a protozoan parasite that causes visceral leishmaniasis. In this study, various in silico, biochemical, biophysical, and cellular assays-based experiments have been conducted to identify and characterize LdTPK, and to provide a sound platform for the discovery of potential LdTPK inhibitors. LdTPK structural modelling ensured high protein quality. Oxythiamine and pyrithiamine were found to bind well with LdTPK with considerable binding energies, and MD simulation-based experiments indicated the stability of the complexation. Additionally, LdTPK1 was found to activate ROS defense in amastigotes, and its inhibition using oxythiamine and pyrithiamine led to the growth inhibition of L. donovani promastigotes and intracellular amastigotes. These findings highlight LdTPK as a promising target for the development of new anti-leishmanial agents. An in-depth analysis of the enzymes involved in TPP biosynthesis in L. donovani has the potential to yield novel therapeutic strategies for Leishmaniasis.Communicated by Ramaswamy H. Sarma.