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BackgroundParamphistomosis is a pathogenic disease of domesticated ruminants, causing great economic loss in dairy industry and meat production. It is considered as a neglected tropical disease with highest prevalence throughout tropical and subtropical regions, particularly in Africa, Asia, Europe, and Australia. There are few trematocidal drugs available in the market. Most are resistant and have elevated side effects. Therefore, alternative trematocidal drugs need to discover. This study was conducted to evaluate three plants leaf extracts (fromSenna alata,S. alexandrina,andS. occidentalis) as effective remedies against gastrointestinal trematode parasite (Paramphistomum gracile) of ruminants.Live adult parasites were collected in 0.1 M phosphate-buffered saline (PBS) from fresh autopsied goat’s rumen. Parasites were incubated in leaf extracts ofS. alata,S. alexandrinaand,S. occidentalisindividually and in combination (1:1) ratio at 37 ± 1°C. Treatment media contain extracts at different concentrations (10, 20 and 40 mg/mL) in 10 mL of 0.1 M PBS with 1% dimethylsulphoxide (DMSO). Parasites in control group were incubated in PBS without extract. The efficacy of threeSennaextracts was evaluated on the basis of dose-dependent motility and mortality of the trematode. Immediately after paralysis, all treated parasites were collected for histology, SEM and biochemical study.ResultsDose-dependent efficacy was observed in terms of motility and time of mortality in all treated parasites after exposure in various concentrations of theSennaplant extracts.S. occidentalisandS. alexandrinashowed better efficacy in combination than comparing with individual treatment groups. Histological study and scanning electron microscopic observations revealed conspicuous deformity of surface architecture in all treated parasites. Scanning electron micrographs also revealed shrinkage, vacuolization, infoldings and blebbings on the body surface of treated worms. Activities of tegumental enzymes were inhibited in all treatment groups compared to control.ConclusionThe overall findings from this study revealed that all threeSennaleaf extracts individually and in combination showed potential antitrematocidal activity againstParamphistomum gracileby damaging body tegument and neural propagation. Thus, this study confirmed that all threeSennaextracts can be considered as a potential drug-like candidate in indigenous system of traditional medicine against trematode infections in livestock.Graphical abstract

Context: Clerodendrum viscosum Vent. (Verbenaceae) is a shrub, widely used amongst the natives of India against various diseases. Objective: Crude extract of the plant was tested in vitro on a tapeworm Raillietina tetragona Molin (Davaineidae) to evaluate its potential anthelmintic efficacy and ultrastructural changes in the parasite. Materials and methods: Parasites were exposed to different concentrations of ethanolic leaf extract (10–80 mg/mL) and praziquantel (0.0005–0.005 mg/mL) and incubated in phosphate-buffered saline (PBS). The pH was 7.4 at 37 °C, while one set of worms was incubated only with PBS as a control. Permanent immobilization of worms was determined visually when no motility occurred on physically disturbing them. The parasites exposed to high concentrations of leaf extract and praziquantel treatments were processed for histological and electron microscopic studies, as these concentrations took the least time for paralysis and death to occur. Result: With an increase in the concentration of the leaf extract from 10 to 80 mg/mL and praziquantel from 0.0005 to 0.005 mg/mL, the time for the onset of paralysis and death was shortened. The treated parasites lost their spontaneous movement rapidly followed by death. Electron microscopic observations revealed disruptions in the tegument and parenchymal layer, accompanied by deformities in cell organelles. Discussion and conclusion: Extensive structural alterations in the tegument indicate that the plant-derived components cause permeability changes in the parasite leading to paralysis and subsequent death. These observations suggest that phytochemicals present in C. viscosum have vermifugal or vermicidal activity, and thus may be exploited as alternative chemotherapeutic agents.

Context Plants and plant products have been used in traditional medicine as anthelmintic agents in human and veterinary medicine. Three species of Senna plant, S. alata (L), S. alexandrina (M) and S. occidentalis (L.) Link (Fabaceae) have been shown to have a vermicidal/vermifugal effect on a zoonotic tapeworm Hymenolepis diminuta (Rudolphi) (Cyclophyllidean). Objective The present study validates the mode of action of these Senna plants on the parasite. The alcoholic leaf extract was determined to obtain information on the intracellular free calcium concentration level. Materials and methods Hymenolepis diminuta was maintained in Sprague-Dawley rat model for 2 months. Live parasites collected from infected rat intestine were exposed to 40 mg/mL concentration of each plant extracts prepared in phosphate buffer saline at 37 °C, till parasite gets paralyzed. The rate of efflux of calcium from the parasite tissue to the medium and the level of intracellular Ca(2+ )concentration were determined by an atomic absorption spectroscopy. Results This study revealed that exposure of the worms to the plant extract leads to disruption in intracellular calcium homeostasis. A significant increase (44.6% and 25%) of efflux in Ca(2+ )from the tissue to the incubated medium was observed. Senna alata showed high rate of efflux (5.32 mg/g) followed by S. alexandria and S. occidentalis (both 4.6 mg/g) compared with control (3.68 mg/g). Discussion and conclusion These results suggest that leaf extracts caused membrane permeability to Ca(2+ )after vacuolization of the tegument under stress and the extracts may contain compound that can be used as a chemotherapeutic agent.

Like mammalian cells, helminth parasites are equipped with an array of enzymatic anti-oxidant system which has an adaptive strategy to cope up with several conditions of stress that arise from host immune response or drug treatment. Earlier, we had reported that three species of Senna, viz. S. alata, S. alexandrina and S. occidentalis leaf extracts caused severe morphological and biochemical alterations in the zoonotic parasite Hymenolepis diminuta. To understand whether the leaf extracts of the three species of Senna have any effect on the enzymatic anti-oxidant system in H.diminuta or not, the present study was investigated on the mechanism of action of these leaf extracts on the anti-oxidant system of the parasite. The viability of the parasite was assessed by MTT reduction assay, chromatin condensation through Hoechst staining of tissue and DNA fragmentation assay, and the oxidative enzymes of the parasite were estimated biochemically. Activity of superoxide dismutase, catalase, glutathione S- transferase and glutathione peroxidase were found to be increased in all the treated parasites from that of the control, with S. alata showed the highest increased amongst the three plant species in all the enzymes, at 331.0 %, 215.4 %, 85.4 % and 65.5 % respectively. Upliftment of apoptotic protein CED-3, CED-4 and EGL-1 and down regulation of anti-apototic protein CED-9 was visualised in all treated paraites. The redox imbalance triggered by these leaf extracts resulted in the activation of apoptotic pathway that led to death of the parasite. Our results demonstrated that the leaf extracts of the three Senna plant species could open new insight for an affordable natural anthelmintic with high efficacy and less toxicity.

Porphyromonas gingivalis is a keystone pathogen that contributes to periodontal pathogenesis by disrupting host-microbe homeostasis and promoting dysbiosis. The virulence of P. gingivalis likely reflects an alteration in the lipid A composition of its lipopolysaccharide (LPS) from the penta-acylated ( Pg LPS 1690 ) to the tetra-acylated ( Pg LPS 1435/1449 ) form. Mast cells play an important role in periodontitis, but the mechanisms of their activation and regulation remain unknown. The expression of epithelium- and neutrophil-derived host defense peptides (HDPs) (LL-37 and human β-defensin-3), which activate mast cells via Mas-related G protein-coupled receptor X2 (MRGPRX2), is increased in periodontitis. We found that MRGPRX2-expressing mast cells are present in normal gingiva and that their numbers are elevated in patients with chronic periodontitis. Furthermore, HDPs stimulated degranulation in a human mast cell line (LAD2) and in RBL-2H3 cells stably expressing MRGPRX2 (RBL-MRGPRX2). Pg LPS 1690 caused substantial inhibition of HDP-induced mast cell degranulation, but Pg LPS 1435/1449 had no effect. A fluorescently labeled HDP (FAM-LL-37) bound to RBL-MRGPRX2 cells, and Pg LPS 1690 inhibited this binding, but Pg LPS 1435/1449 had no effect. These findings suggest that low-level inflammation induced by HDP/MRGPRX2-mediated mast cell degranulation contributes to gingival homeostasis but that sustained inflammation due to elevated levels of both HDPs and MRGPRX2-expressing mast cells promotes periodontal disease. Furthermore, differential regulation of HDP-induced mast cell degranulation by Pg LPS 1690 and Pg LPS 1435/1449 may contribute to the modulation of disease progression.

The survival of intracellular protozoan parasite, Leishmania donovani, the causative agent of Indian visceral leishmaniasis (VL), depends on the activation status of macrophages. l-Arginine, a semi-essential amino acid plays a crucial regulatory role for activation of macrophages. However, the role of l-arginine transport in VL still remains elusive. In this study, we demonstrated that intra-macrophage survival of L. donovani depends on the availability of extracellular l-arginine. Infection of THP-1-derived macrophage/human monocyte-derived macrophage (hMDM) with Leishmania, resulted in upregulation of l-arginine transport. While investigating the involvement of the transporters, we observed that Leishmania survival was greatly impaired when the transporters were blocked either using inhibitor or siRNA-mediated downregulation. CAT-2 was found to be the main isoform associated with l-arginine transport in L. donovani-infected macrophages. l-arginine availability and its transport regulated the host arginase in Leishmania infection. Arginase and inducible nitric oxide synthase (iNOS) expression were reciprocally regulated when assayed using specific inhibitors and siRNA-mediated downregulation. Interestingly, induction of iNOS expression and nitric oxide production were observed in case of inhibition of arginase in infected macrophages. Furthermore, inhibition of l-arginine transport as well as arginase resulted in decreased polyamine production, limiting parasite survival inside macrophages. l-arginine availability and transport regulated Th1/Th2 cytokine levels in case of Leishmania infection. Upregulation of l-arginine transport, induction of host arginase, and enhanced polyamine production were correlated with increased level of IL-10 and decreased level of IL-12 and TNF-α in L. donovani-infected macrophages. Our findings provide clear evidence for targeting the metabolism of l-arginine and l-arginine-metabolizing enzymes as an important therapeutic and prophylactic strategy to treat VL.

Visceral leishmaniasis is a vector-borne disease caused by an obligate intracellular protozoan parasite Leishmania donovani. The molecular mechanism involved in internalization of Leishmania is poorly understood. The entry of Leishmania involves interaction with the plasma membrane of host cells. We have previously demonstrated the requirement of host membrane cholesterol in the binding and internalization of L. donovani into macrophages. In the present work, we explored the role of the host actin cytoskeleton in leishmanial infection. We observed a dose-dependent reduction in the attachment of Leishmania promastigotes to host macrophages upon destabilization of the actin cytoskeleton by cytochalasin D. This is accompanied by a concomitant reduction in the intracellular amastigote load. We utilized a recently developed high resolution microscopy-based method to quantitate cellular F-actin content upon treatment with cytochalasin D. A striking feature of our results is that binding of Leishmania promastigotes and intracellular amastigote load show close correlation with cellular F-actin level. Importantly, the binding of Escherichia coli remained invariant upon actin destabilization of host cells, thereby implying specific involvement of the actin cytoskeleton in Leishmania infection. To the best of our knowledge, these novel results constitute the first comprehensive demonstration on the specific role of the host actin cytoskeleton in Leishmania infection. Our results could be significant in developing future therapeutic strategies to tackle leishmaniasis.

Objective: To study the in vitro anthelmintic efficacy of Cassia alata (C. alata), Cassia angustifolia (C. angustifolia) and Cassia occidentalis (C. occidentalis). Methods: Crude ethanol extract from leaves of the three plants were prepared in rotary evaporator and different concentrations (10, 20 and 40 mg/mL) of leaf extracts were used for treatment on different representatives of helminthes (Heterakis gallinarum, Raillietina tetragona and Catatropis sp.) from domestic fowl (Gallus gallus domesticus). Loss of motility and death were monitored frequently. Results: C. alata showed early paralysis in all worms treated followed by C. angustifolia. C. occidentalis in combination with C. alata together caused early paralysis in all treated worms than the combination of C. alata with C. angustfolia. While Heterakis gallinarum in control survived for (81.33 ± 2.07) h, treated worms lost their motility at (5.71 ± 0.10) h, (6.60 ± 0.86) h and (13.95 ± 0.43) h with C. angustifolia, C. alata and C. occidentalis respectively at a concentration of 40 mg/mL which showed better efficacy than albendazole. Catatropis sp. survival period was (26.49 ± 1.38) h in control, but with plant treatment, it lost its motility in just (0.57 ± 0.08) h, (1.00 ± 0.12) h and (1.47 ± 0.40) h at 40 mg/mL concentration of C. alata, C. angustifolia and C. occidentalis respectively. Raillietina tetragona on the other hand became paralysed at (1.68 ± 0.27) h, (2.95 ± 0.29) h and (4.13 ± 0.31) h with above concentrations treated with three plants respectively, however in control it survived up to (81.93 ± 4.71) h. Conclusions: This present study indicated broad spectrum vermifugal activity of all plants tested.

Background Leishmania donovani, belonging to a unicellular protozoan parasite, display the differential level of linkage-specific sialic acids on their surface. Sialic acids binding immunoglobulin-like lectins (siglecs) are a class of membrane-bound receptors present in the haematopoetic cell lineages interact with the linkage-specific sialic acids. Here we aimed to explore the utilization of sialic acids by Leishmania donovani for siglec-mediated binding, phagocytosis, modulation of innate immune response and signaling pathways for establishment of successful infection in the host. Methodology/Principle Findings We have found enhanced binding of high sialic acids containing virulent strains (AG83+Sias) with siglec-1 and siglec-5 present on macrophages compared to sialidase treated AG83+Sias (AG83-Sias) and low sialic acids-containing avirulent strain (UR6) by flow cytometry. This specific receptor-ligand interaction between sialic acids and siglecs were further confirmed by confocal microscopy. Sialic acids-siglec-1-mediated interaction of AG83+Sias with macrophages induced enhanced phagocytosis. Additionally, sialic acids-siglec-5 interaction demonstrated reduced ROS, NO generation and Th2 dominant cytokine response upon infection with AG83+Sias in contrast to AG83-Sias and UR6. Sialic acids-siglecs binding also facilitated multiplication of intracellular amastigotes. Moreover, AG83+Sias induced sialic acids-siglec-5-mediated upregulation of host phosphatase SHP-1. Such sialic acids-siglec interaction was responsible for further downregulation of MAPKs (p38, ERK and JNK) and PI3K/Akt pathways followed by the reduced translocation of p65 subunit of NF-κβ to the nucleus from cytosol in the downstream signaling pathways. This sequence of events was reversed in AG83-Sias and UR6-infected macrophages. Besides, siglec-knockdown macrophages also showed the reversal of AG83+Sias infection-induced effector functions and downstream signaling events. Conclusions/Significances Taken together, this study demonstrated that virulent parasite (AG83+Sias) establish a unique sialic acids-mediated binding and subsequent phagocytosis in the host cell through the selective exploitation of siglec-1. Additionally, sialic acids-siglec-5 interaction altered the downstream signaling pathways which contributed impairment of immune effector functions of macrophages. To the best of our knowledge, this is a comprehensive report describing sialic acids-siglec interactions and their role in facilitating uptake of the virulent parasite within the host., Author Summary Sialic acids are nine carbon sugars present on terminal cell surface glycoproteins and glycolipids. Siglec is a membrane receptor that belongs to an immunoglobulin super family present in almost all the haematopoetic cell lineages. There are 14 different types of siglecs present on human immune cells that take an active part in balancing the magnitude of immunological reactions. In general, these siglecs bind with sialic acids and negatively regulate the immune response. Leishmania contains sialic acids on its surface. Virulent parasites utilize this sugar to bind with macrophages through siglec-1 and siglec-5 compared to low sialic acids containing avirulent parasites. Such sialic acids-siglec-mediated interactions exhibited a suppressed host immune response which helped them to establish successful infection compared to desialylated virulent and avirulent parasites, as well as, siglec-depleted macrophages. Interestingly, interaction between sialic acids and siglec-1 induced enhanced phagocytosis, while sialic acids-siglec-5 interaction upregulated the phosphatase SHP-1. This interaction with the virulent strain exhibited deactivation of various downstream signaling pathways and ultimately controlled translocation of a functional component of transcription factor NF-κβ for regulation of cytokines and other effector molecules in infected macrophages. Thus, the interaction between the parasite and the host cells through sialic acids-siglec binding is clearly a newly identified mechanism by which parasites can establish successful infection by subverting the host’s innate immune response.

Senna alexandrina Mill. has been used for antimicrobial activity. In the present study, the crude ethanolic extract of the plant and a synthetic compound Sennoside were tested in vitro on Hymenolepis diminuta to evaluate its potential anthelmintic efficacy through ultrastructural changes. Worms were maintained between rat model and beetle and the test parasites were exposed to different concentrations of crude ethanolic leaf extracts of S. alexandrina. Praziquantel was used as a reference drug. Dose dependent efficacy was observed in terms of motility and time of mortality in all treated parasites. Ultrastructural micrography revealed irrevocable destruction all over the body tegument accompanied with sloughing of microtriches and swellings of the basal lamina. Vacuolization of the syncytium along with sparsely cytoplasmic cytons and depletion of parenchymatous layer were observed accompanied by deformities in the cell organelles. Extensive deformities in the tegument indicates that the plant extract alter membrane permeability of the parasite leading to paralysis and subsequent death. Thus, S. alexandrina can be regarded as a potential anthelmintic agent.

Leishmania are obligate intracellular protozoan parasites that invade and survive within host macrophages leading to leishmaniasis, a major cause of mortality and morbidity worldwide, particularly among economically weaker sections in tropical and subtropical regions. Visceral leishmaniasis is a potent disease caused by Leishmania donovani. The detailed mechanism of internalization of Leishmania is poorly understood. A basic step in the entry of Leishmania involves interaction of the parasite with the host plasma membrane. In this work, we have explored the effect of chronic metabolic cholesterol depletion using lovastatin on the entry and survival of Leishmania donovani in host macrophages. We show here that chronic cholesterol depletion of host macrophages results in reduction in the attachment of Leishmania promastigotes, along with a concomitant reduction in the intracellular amastigote load. These results assume further relevance since chronic cholesterol depletion is believed to mimic physiological cholesterol modulation. Interestingly, the reduction in the ability of Leishmania to enter host macrophages could be reversed upon metabolic replenishment of cholesterol. Importantly, enrichment of host membrane cholesterol resulted in reduction in the entry and survival of Leishmania in host macrophages. As a control, the binding of Escherichia coli to host macrophages remained invariant under these conditions, thereby implying specificity of cholesterol requirement for effective leishmanial infection. To the best of our knowledge, these results constitute the first comprehensive demonstration that an optimum content of host membrane cholesterol is necessary for leishmanial infection. Our results assume relevance in the context of developing novel therapeutic strategies targeting cholesterol-mediated leishmanial infection.

Withania somnifera (Ashwagandha) is a plant with known ethnomedicinal properties and its use in Ayurvedic medicine in India is well documented. The present investigation reports on immunomodulatory efficacy of aqueous-ethanol extracts of roots of three selected Withania somnifera chemotypes designated as NMITLI 101R, NMITLI 118R and NMITLI 128R. Each chemotype was administered 10–100 mg/kg orally to BALB/c mice once daily for 14 days. The immunomodulatory consequences were recorded by determining the humoral immune response with the help of hemagglutination, plaque forming cell assay and cellular response by measuring delayed type hypersensitivity reaction. Additionally, other immune parameters such as proliferation of T and B cells, intracellular and secreted Th1 and Th2 cytokines along with modulation in ROS production by peritoneal macrophages were monitored after feeding with lower doses (3–30 mg/kg/day) of these three chemotypes individually. NMITLI 101R incited both humoral and cellular immune response in terms of higher number of antibody producing cells and enhanced foot pad swelling at the 10 mg dose as also dose dependent B and T cell proliferations. Levels of intracellular and secreted cytokines post-NMITLI 101R treatment illustrated generation of mixed Th1/Th2 response that remained more polarized towards Th1. This chemotype also generated maximum reactive oxygen species. NMITLI 118R provoked comparatively reduced immune response in all humoral and cellular parameters at lower doses but induced highly polarized Th1 cytokine response. In contrast, NMITLI 128R led to enhanced antibody production with minimal cellular response demonstrating marginally Th2 dominance at a lower dose. Taken together, it may therefore be concluded that there were distinct modulation in the immune response exhibited by the three chemotypes of Withania somnifera and NMITLI 101R appeared to possess a better immunostimulatory activity than the other chemotypes at lower doses.

Objective: The present study aims at observing the effects of three species of Senna plants, viz. Senna alata , S. alexandrina and S. occidentalis on the ultrastructure of the mitochondria of the tapeworm, Hymenolepis diminuta . Methods: Worms were treated with leaf extracts of the three plant species with a standard dose concentration of 40 mg/ml and keeping one group of parasites in phosphate buffer saline (PBS) as a control. The parasites from control and treated medium were simultaneously removed after the loss of motility and fixed in 3% gluteraldehyde. They were processed for ultramicrograph observations of the worm’s mitochondria with special reference to shape and cytoplasm through transmission electron microscopy (TEM). Results: The study showed loss of architecture in the outer mitochondrial membrane. The inner membrane became distorted with inconspicuous cristae and matrix became lucent in all plant treated worms compared to control. Amongst the three plants, S. alexandrina showed overall distortion in the shape leading to bloating of mitochondria. Conclusion: The observations depict pronounced alterations in the structure of mitochondria, thus signifying depletion of energy synthesis in the parasite. Senna plant could, therefore, be a potent anthelmintic alternative.

Queen is the most preferred table purpose variety in India. However, small fruit size, deep set eyes in large numbers and weak fruit stalk make this less preferred in processing industries. A completely different plant type with distinct fruit character was identified from a clonal population of Queen which seems to be a natural mutant. The identified mutant was maintained and multiplied through vegetative propagation at Lembucherra farm of ICAR Research Complex, Tripura centre. The present study clearly established the fact that this new clone (PQM-1) is distinct from 'queen' and other cultivars of Tripura. Distinctness of this new clone was confirmed on the basis of standard morphological and biochemical descriptors as well as through DNA polymorphism. It was observed that PQM-1 diverged from Queen in respect of pattern of vegetative growth, leaf shape, size and colour, flowering time, fruit shape, size and colour, eye shape, fruit weight, juice content, total soluble solid (TSS) and acidity. PQM-1 was found uniform at DNA level and complement the information of phenotypic and quality parameters by showing stable differences with Queen in ISSR banding pattern. Natural mutation in DNA level might be the probable cause of its origin. It could be empathetically stated that PQM-1 would have certain competitive advantage over all existing cultivars in this region due to its late maturity, fruit quality and usefulness in both canning and fresh consumption. This certainly would emerge as new alternative to pineapple growers of this region.

The beneficial effects of two probiotic bacteria, Bacillus licheniformis and Bacillus megaterium, on growth, biochemical, and immunological responses of Litopenaeus vannamei (Boone, 1931) were evaluated both individually and in combination. A 60 day experimental trial was conducted on 1200 post larvae (PL) of Litopenaeus vannamei divided into four experimental groups, with three replicates of each treatment, and 100 PL in each replicate. The shrimp were fed a commercial diet (32% protein) supplemented with different probiotics. The first treatment included only B. licheniformis @ ~ 1x 108 cfu/kg diet (T1), the second consisted of a combination of ~ 0.5 x 108 cfu/kg of B. licheniformis mixed with a similar concentration of B. megaterium (T2), and the third had only B. megaterium @ ~1x 108 cfu/kg diet (T3). The control diet (C) had neither of the probiotic bacteria. Growth performance, biochemical, and immunological parameters of experimental shrimp were measured at 20 day intervals. Probiotic fed shrimp showed significantly higher growth, biochemical, and immunological response than the control. Among the probiotic treatments the mixture of B. licheniformis and B. megaterium (T2) performed better than the individual probiotics alone, indicating the synergistic effect of these two functionally different probiotics

Senna alata has attracted the attention of many researchers due to its numerous medicinal properties. This study aims to test the acute and sub-acute toxicity of its leaf extracts in Swiss albino mice. Studies were carried out with a fixed dose of 1,000, 2,000, and 3,000 mg/kg body weight through oral administration daily. Signs of toxicity in terms of behavior and mortality were noted after every two hours till 24 h of administration for acute toxicity and further administration of extracts till 15 days to analyze the physical, biochemical, hematological parameters, and histopathological studies in liver, kidney, and spleen for sub-acute study. The highest dose administered did not produce mortality or changes in the general behavior of the test animals. All parameters were unaltered throughout the study. The present study revealed no obvious toxicity in mice treated with S. alata. These results indicate the safety of the oral administration of leaf extract.

The growth and survival of intracellular parasites depends on the availability of extracellular nutrients. Deprivation of nutrients viz glucose or amino acid alters redox balance in mammalian cells as well as some lower organisms. To further understand the relationship, the mechanistic role of L-arginine in regulation of redox mediated survival of Leishmania donovani promastigotes was investigated. L-arginine deprivation from the culture medium was found to inhibit cell growth, reduce proliferation and increase L-arginine uptake. Relative expression of enzymes, involved in L-arginine metabolism, which leads to polyamine and trypanothione biosynthesis, were downregulated causing decreased production of polyamines in L-arginine deprived parasites and cell death. The resultant increase in reactive oxygen species (ROS), due to L-arginine deprivation, correlated with increased NADP+/NADPH ratio, decreased superoxide dismutase (SOD) level, increased lipid peroxidation and reduced thiol content. A deficiency of L-arginine triggered phosphatidyl serine externalization, a change in mitochondrial membrane potential, release of intracellular calcium and cytochrome-c. This finally led to DNA damage in Leishmania promastigotes. In summary, the growth and survival of Leishmania depends on the availability of extracellular L-arginine. In its absence the parasite undergoes ROS mediated, caspase-independent apoptosis-like cell death. Therefore, L-arginine metabolism pathway could be a probable target for controlling the growth of Leishmania parasites and disease pathogenesis.

The efficacy of Cassia alata against cestode Hymenolepis diminuta was evaluated in our study. Worms were maintained between rat model and beetle. Air-dried leaves of C. alata were subjected to ethanol extraction. Different concentrations (10–80 mg/ml) of the extract were prepared in phosphate buffer saline and tested on the parasite in vitro. Parasites treated with C. alata showed a decrease in motility with an increase in concentrations and complete immobilization took lesser time compared to control. The paralyzed parasites were further processed for electron microscopic studies. Ultrastructural micrographs revealed swelling of the tegument and blebbing on the tegumental surface throughout the body accompanied with destruction of microtriches and changes such as shrinkage in the scolex region. Depletion of parenchyma cells and destruction in the connective tissues along with sparsely cytoplasmic cytons were also observed, and these observations are similar with worms treated with a known drug praziquantel. These results may suggest that the plant leaves could be considered for controlling helminth infection and can represent a step forward in the search for alternative anthelmintic drug.

During cancer, immune system gets suppressed due to developing malignancy and anticancer drugs used. Therefore, an ideal chemotherapeutic agent would have both immuno-stimulatory and cancer cell death-inducing activity. Here, we have shown that Withanolide D (WithaD), an important bioactive withanolide, purified from the leaves of an Indian chemotype (NMITLI 135) of Withania somnifera acts as stimulator of Th1 response along with moderate effect on Th2 response, thus advocates its candidature as a potent immunostimulator. Additionally, WithaD exhibited antileukemic activity, targeting multiple pathways along with ceramide accumulation through N-SMase 2 activation, ultimately inducing apoptosis in neoplastic cells. Taken together, WithaD is a potential herbal compound with dual functions: potentiation of the intrinsic defense mechanism and inducer of cancer cell apoptosis.