Objective: To evaluate the antimalarial activity of noscapine against Plasmodium falciparum 3D7 strain (Pf3D7), its clinical isolate (Pf140/SS), and Plasmodium berghei ANKA (PbA). Methods: Using ring-stage survival assay, phenotypic assessments, and SYBR-green-based fluorescence assay, the antimalarial activities of noscapine were assessed compared with dihydroartemisinin (DHA) in in vivo and in vitro studies. In addition, hemolysis and cytotoxicity tests were carried out to evaluate its safety. RT-PCR assay was also conducted to determine the effect of noscapine on papain-like cysteine protease Plasmodium falciparum falcipain-2 (PfFP-2). Results: The antimalarial efficacy of noscapine against Pf3D7 and Pf140/SS was comparable to DHA, with IC50 values of (7.68±0.88) and (5.57±0.74) nM/mL, respectively, and >95% inhibition of PbA infected rats. Noscapine also showed a safe profile, as evidenced by low hemolysis and cytotoxicity even at high concentrations. Moreover, PfFP-2 expression was significantly inhibited in both noscapine-treated Pf3D7 and Pf140/SS (P<0.01). Conclusions: Noscapine has antimalarial properties comparable to standard antimalarial DHA with better safety profiles, which may be further explored as a therapeutic candidate for the treatment of malaria.

Malaria parasite lacks canonical pathways for amino acid biosynthesis and depends primarily on hemoglobin degradation and extracellular resources for amino acids. Interestingly, a putative gene for glutamine synthetase (GS) is retained despite glutamine being an abundant amino acid in human and mosquito hosts. Here we show Plasmodium GS has evolved as a unique type I enzyme with distinct structural and regulatory properties to adapt to the asexual niche. Methionine sulfoximine (MSO) and phosphinothricin (PPT) inhibit parasite GS activity. GS is localized to the parasite cytosol and abundantly expressed in all the life cycle stages. Parasite GS displays species-specific requirement in Plasmodium falciparum (Pf) having asparagine-rich proteome. Targeting PfGS affects asparagine levels and inhibits protein synthesis through eIF2α phosphorylation leading to parasite death. Exposure of artemisinin-resistant Pf parasites to MSO and PPT inhibits the emergence of viable parasites upon artemisinin treatment.

Hematological abnormalities such as anemia and coagulopathies are well-established complications in malaria. Other well-recognized complications of malaria are cerebral malaria and kidney injury. In this case report, we present a man that presented with a gastrointestinal bleeding as a complication of a malaria infection, which was found through histopathological investigations of the resected colon.

Abstract Malaria is a global public health menace. The quest for new antimalarials and adjuvants for malaria in the backdrop of artemisinin resistance has been enormous. This study evaluates the comprehensive antimalarial activity of the natural phytochemical compound Noscapine, against Plasmodium falciparum 3D7 strain (Pf3D7), clinical isolate (Pf140/SS) and in Plasmodium berghei ANKA (PbA), inhibiting in vitro and in vivo parasite growth under controlled conditions as evaluated through the ring-stage survival assay, phenotypic assessments and SYBR-green based fluorescence assay. Cytotoxicity of Noscapine was evaluated against the J774. A.1 murine macrophage cell line besides profiling its hemolysis activities against human RBCs. The antimalarial efficacy of Noscapine against Pf3D7 and Pf140/SS was similar or better than standard antimalarial Dihydroartemisinin (DHA), with the IC50 value of 7.68±0.88 and 5.57±0.74 nM/mL respectively along with more than 95% inhibition in infected Wister albino rats with PbA after 4-day suppressive test. Importantly, unlike DHA, no toxicity symptoms were observed with CC50 value 1748 nM/mL or hemolysis with Nospcapine, even at extremely high concentrations. Based on the published literature as on date, this is the first report on native Noscapine, which has shown potent antimalarial efficacy and safety profiles vis-à-vis standard antimalarial DHA, demonstrated through in vitro and in vivo models of animal and clinical malaria parasites.

Limited availability of the organs donors has facilitated the establishment of xenogeneic organ sources for transplantation. Numerous studies have decellularized several organs and assessed their implantability in order to provide such organs. Among all the decellularized organs studies for xenotransplantation, the pancreas has garnered very limited amount of research. The presently offered alternatives for pancreas transplantation are unable to liberate patients from donor dependence. The rat and mice pancreas are not of an accurate size for transplantation but can only be used for in-vitro studies mimicking in-vivo immune response in humans, while the porcine pancreas can cause zoonotic diseases as it carries porcine endogenous retrovirus (PERV- A/B/C). Therefore, we propose caprine pancreas as a substitute for these organs, which not only reduces donor dependence but also poses no risk of zoonosis. Upon decellularization the extracellular matrix (ECM) of different tissues responds differently to the detergents used for decellularization at physical and physiological level; this necessitates a comprehensive analysis of each tissue independently. This study investigates the impact of decellularization by ionic (SDS and SDC), non-ionic (Triton X-100 and Tween-20), and zwitterionic detergents (CHAPS). All these five detergents have been used to decellularize caprine pancreas via immersion (ID) and perfusion (PD) set-up. In this study, an extensive comparison of these two configurations (ID and PD) with regard to each detergent has been conducted. The final obtained scaffold with each set-up has been evaluated for the left-over cytosolic content, ECM components like sGAG, collagen, and fibronectin were estimated via Prussian blue and Immunohistochemical staining respectively, and finally for the tensile strength and antimicrobial activity. All the detergents performed consistently superior in PD than in ID. Conclusively, PD with SDS, SDC, and TX-100 successfully decellularizes caprine pancreatic tissue while retaining ECM architecture and mechanical properties. This research demonstrates the viability of caprine pancreatic tissue as a substitute scaffold for porcine organs and provides optimal decellularization protocol for this xenogeneic tissue. This research aims to establish a foundation for further investigations into potential regenerative strategies using this ECM in combination with other factors.

In this study, the caprine pancreas has been presented as an alternative to the porcine organ for pancreatic xenotransplantation with lesser risk factors. The obtained caprine pancreas underwent a systematic cycle of detergent perfusion for decellularization. It was perfused using anionic (0.5% w/v sodium dodecyl sulfate) as well as non-ionic (0.1% v/v triton X-100, t-octyl phenoxy polyethoxy ethanol) detergents and washed intermittently with 1XPBS supplemented with 0.1% v/v antibiotic and nucleases in a gravitation-driven set-up. After 48 h, a white decellularized pancreas was obtained, and its extracellular matrix (ECM) content was examined for scaffold-like properties. The ECM content was assessed for removal of cellular content, and nuclear material was evaluated with temporal H&E staining. Quantified DNA was found to be present in a negligible amount in the resultant decellularized pancreas tissue (DPT), thus prohibiting it from triggering any immunogenicity. Collagen and fibronectin were confirmed to be preserved upon trichrome and immunohistochemical staining, respectively. SEM and AFM images reveal interconnected collagen fibril networks in the DPT, confirming that collagen was unaffected. sGAG was visualized using Prussian blue staining and quantified with DMMB assay, where DPT has effectively retained this ECM component. Uniaxial tensile analysis revealed that DPT possesses better elasticity than NPT (native pancreatic tissue). Physical parameters like tensile strength, stiffness, biodegradation, and swelling index were retained in the DPT with negligible loss. The cytocompatibility analysis of DPT has shown no cytotoxic effect for up to 72 h on normal insulin-producing cells (MIN-6) and cancerous glioblastoma (LN229) cells in vitro. The scaffold was recellularized using isolated mouse islets, which have established in vitro cell proliferation for up to 9 days. The scaffold received at the end of the decellularization cycle was found to be non-toxic to the cells, retained biological and physical properties of the native ECM, suitable for recellularization, and can be used as a safer and better alternative as a transplantable organ from a xenogeneic source.

Background: National Malaria Control Programmes (NMCPs) currently make limited use of parasite genetic data. We have developed GenRe-Mekong, a platform for genetic surveillance of malaria in the Greater Mekong Subregion (GMS) that enables NMCPs to implement large-scale surveillance projects by integrating simple sample collection procedures in routine public health procedures. Methods: Samples from symptomatic patients are processed by SpotMalaria, a high-throughput system that produces a comprehensive set of genotypes comprising several drug resistance markers, species markers and a genomic barcode. GenRe-Mekong delivers Genetic Report Cards, a compendium of genotypes and phenotype predictions used to map prevalence of resistance to multiple drugs. Results: GenRe-Mekong has worked with NMCPs and research projects in eight countries, processing 9623 samples from clinical cases. Monitoring resistance markers has been valuable for tracking the rapid spread of parasites resistant to the dihydroartemisinin-piperaquine combination therapy. In Vietnam and Laos, GenRe-Mekong data have provided novel knowledge about the spread of these resistant strains into previously unaffected provinces, informing decision-making by NMCPs. Conclusions: GenRe-Mekong provides detailed knowledge about drug resistance at a local level, and facilitates data sharing at a regional level, enabling cross-border resistance monitoring and providing the public health community with valuable insights. The project provides a rich open data resource to benefit the entire malaria community. Funding: The GenRe-Mekong project is funded by the Bill and Melinda Gates Foundation (OPP11188166, OPP1204268). Genotyping and sequencing were funded by the Wellcome Trust (098051, 206194, 203141, 090770, 204911, 106698/B/14/Z) and Medical Research Council (G0600718). A proportion of samples were collected with the support of the UK Department for International Development (201900, M006212), and Intramural Research Program of the National Institute of Allergy and Infectious Diseases.

Cerebral malaria (CM) is caused by the binding of Plasmodium falciparum-infected erythrocytes (IEs) to the brain microvasculature, leading to inflammation, vessel occlusion, and cerebral swelling. We have previously linked dual intercellular adhesion molecule-1 (ICAM-1)- and endothelial protein C receptor (EPCR)-binding P. falciparum parasites to these symptoms, but the mechanism driving the pathogenesis has not been identified. Here, we used a 3D spheroid model of the blood-brain barrier (BBB) to determine unexpected new features of IEs expressing the dual-receptor binding PfEMP1 parasite proteins. Analysis of multiple parasite lines shows that IEs are taken up by brain endothelial cells in an ICAM-1-dependent manner, resulting in breakdown of the BBB and swelling of the endothelial cells. Via ex vivo analysis of postmortem tissue samples from CM patients, we confirmed the presence of parasites within brain endothelial cells. Importantly, this discovery points to parasite ingress into the brain endothelium as a contributing factor to the pathology of human CM.

Background The role of microbiota in the pathophysiology of benign prostate hyperplasia (BPH), especially in creating an inflammatory milieu may not be avoided. The major objectives of this study were to investigate the microbial composition of BPH tissues, its association with inflammation and check the effect of clinically isolated bacteria on prostate epithelial cells. Methods The study includes 36 patients with a pathological diagnosis of BPH. Following strict aseptic measures, tissues were collected after transurethral resection of prostate, multiple pieces of the resected tissues were subjected to histopathological analysis, bacterial culture and genomic DNA extraction. Microbial composition was analyzed by culture and/or next-generation sequencing methods. Annotation of operational taxonomy unit has been done with an in-house algorithm. The extent of inflammation was scored through histological evaluation of tissue sections. The effect of clinical isolates on nuclear factor-kappa B (NF-kappa B) activity and induction of DNA-damage in the prostate epithelial cells were evaluated. Results Histopathological analysis of the BPH tissues showed the presence of inflammation in almost all the tissues with a varied level at different regions of the same tissue section and the level of overall inflammation was different from patients to patients. Microbial culture of tissue samples showed the presence of live bacteria in 55.5% (20 out of 36) of the patient tissues. Majority of the isolates were coagulase-positiveStaphylococcus, E. coliandMicrococcus spp. Further, V3 16S rRNA sequencing of the DNA isolated from BPH tissues showed the presence of multiple bacteria and the most common phylum in the BPH tissues were found to beProteobacteria, Actinobacteria, Firmicutes, andBacteroidetes. TheE. coli, isolated from one of the tissue was able to activate NF-kappa B and induce DNA damage in prostate epithelial cells. Phospho-histone gamma H2A.X staining confirmed the presence of cells with damaged DNA lesion in BPH tissues and also correlated with the severity of inflammation. Conclusion Our study has shown that the BPH tissues do have a divergent microbial composition including the commonly foundE. coli(phylum Proteobacteria), and these bacteria might contribute to the BPH-associated inflammation and/or tissue damage. The BPH-associatedE. coliinduced NF-kappa B signaling and DNA damage in prostate epithelial cells in vitro.

Plasmodium falciparum gametocytes, the sexual stage responsible for malaria parasite transmission from humans to mosquitoes, are key targets for malaria elimination. Immature gametocytes develop in the human bone marrow parenchyma, where they accumulate around erythroblastic islands. Notably though, the interactions between gametocytes and this hematopoietic niche have not been investigated. Here, we identify late erythroblasts as a new host cell for P falciparum sexual stages and show that gametocytes can fully develop inside these nucleated cells in vitro and in vivo, leading to infectious mature gametocytes within reticulocytes. Strikingly, we found that infection of erythroblasts by gametocytes and parasite-derived extracellular vesicles delay erythroid differentiation, thereby allowing gametocyte maturation to coincide with the release of their host cell from the bone marrow. Taken together, our findings highlight new mechanisms that are pivotal for the maintenance of immature gametocytes in the bone marrow and provide further insights on how Plasmodium parasites interfere with erythropoiesis and contribute to anemia in malaria patients.

BackgroundThe control of malaria in pregnancy (MiP) in India relies on testing women who present with symptoms or signs suggestive of malaria. We hypothesised that intermittent screening and treatment for malaria at each antenatal care visit (ISTp) would improve on this approach and reduce the adverse effects of MiP.MethodsA cluster randomised controlled trial comparing ISTp versus passive case detection (PCD) was conducted in Jharkhand state. Pregnant women of all parities with a gestational age of 18–28 weeks were enrolled. Women in the ISTp group were screened with a rapid diagnostic test (RDT) for malaria at each antenatal clinic visit and those in the PCD group were screened only if they had symptoms or signs suggestive of malaria. All RDT positive women were treated with artesunate/sulfadoxine–pyrimethamine. The primary endpoint was placental malaria, determined by placental histology, and the key secondary endpoints were birth weight, gestational age, vital status of the newborn baby and maternal anaemia.ResultsBetween April 2012 and September 2015, 6868 women were enrolled; 3300 in 46 ISTp clusters and 3568 in 41 PCD clusters. In the ISTp arm, 4.9% of women were tested malaria positive and 0.6% in the PCD arm. There was no difference in the prevalence of placental malaria in the ISTp (87/1454, 6.0%) and PCD (65/1560, 4.2%) groups (6.0% vs 4.2%; OR 1.34, 95% CI 0.78 to 2.29, p=0.29) or in any of the secondary endpoints.ConclusionISTp detected more infections than PCD, but monthly ISTp with the current generation of RDT is unlikely to reduce placental malaria or impact on pregnancy outcomes. ISTp trials with more sensitive point-of-care diagnostic tests are needed.

A field experiment was conducted at the experimental field site of All India co-ordinated Research project on vegetable crops, Orissa University of Agriculture Technology, Bhubaneswar in RBD with 7 treatments replicated 3 times during Kharif, 2016 and summer, 2017, to study the field efficacy of six sprayable insecticides viz., rynaxypyr 20% SC @ 33.33 ml a.i/ha, spinosad 45 % SC @ 75 g a.i./ha, emamectin benzoate 5%SG @ 10 g a.i/ha, flubendiamide 480 SC @ 78.70 g a.i. /ha, thiamethoxam 25%WG @ 41.66 g a.i/ha and cartap hydrochloride 50SP@ 375g a..i/ha against brinjal shoot and fruit borer. All the test insecticides proved their efficacy but flubendiamide 480 SC @ 78.70 g a.i. /ha, rynaxypyr 20% SC @ 33.33 ml a.i/ha and spinosad 45%SC @ 10ml a.i/ha proved highly effective against shoot and fruit borer.

BackgroundPrevious studies have shown the effect of bacterial lipopolysaccharide (LPS) on enhanced cancer cells’ growth and metastasis. However, the effect of LPS on prostate cancer (PCa) cells metastasis has not been investigated in details. This study aimed to investigate the functional role of LPS on PCa cells metastasis and determine the effect of dexamethasone (DEX) on this event.MethodsTwo different PCa reporter cells lines (DU145‐NF‐κB‐Luc and MAT‐LyLu‐ NF‐κB‐Luc) were used to assess the direct effect of LPS on NF‐κB activation in PCa cells. Plasma collected from LPS‐stimulated human and rodent blood were used to check the indirect effect of LPS on NF‐κB activation in PCa cells. Trans‐well migration assay and two different orthotopic PCa animal models were used to investigate the effect of LPS on DU145 and MAT‐LyLu cells migration or metastasis in vitro and in vivo, respectively. In all the studies DEX was used with or without LPS stimulation.ResultsLPS and secretory factors present in plasma collected from LPS‐stimulated blood, significantly activated NF‐κB in DU145, and MAT‐LyLu cells and enhanced their migration in vitro. DEX significantly suppressed LPS‐mediated activation of cancer and blood cells and abrogated the direct and indirect pro‐migratory effect of LPS on PCa cells. Systemic administration of LPS activated NF‐κB in DU145 cells in vivo; however, failed to alter the metastatic properties of these cells. On the other hand, systemic administration of LPS to MAT‐LyLu tumor bearing animals significantly enhanced the incidence of metastasis without altering the overall growth of primary tumors. Unexpectedly, though DEX significantly suppressed MAT‐LyLu primary tumor weights, it aggravated metastasis of cancer cells in presence and absence of LPS. Moreover, consecutive DEX pre‐treatment enhanced experimental peritoneal metastasis of MAT‐LyLu cells. At the molecular level, LPS, and/or DEX induced overexpression of immunosuppressive molecules in MAT‐LyLu tumors.ConclusionsOverall, our study has shown that LPS and/or LPS induced inflammation can increase PCa metastasis and immunosuppressive dose of DEX might further enhance cancer metastasis.

Abstract Background: Rapid and accurate diagnosis is crucial in the treatment of malaria. Rapid Diagnostic Tests (RDTs) using blood have been recommended by the WHO as an acceptable method for the diagnosis of malaria. RDTs provide results quickly, is simple to use and easy to interpret. However, its use requires collection of blood by skin puncture. Hence the aim of the pilot study is to explore the sensitivity and specificity of RDTs using urine (collected non-invasively) for diagnosis of Plasmodium falciparum malaria and to assess the relation between parasite density in blood with HRP-2 Ag detection in urine. Material and Method: All fever cases admitted to Ispat General Hospital (IGH) Rourkela, India, during June 2012-March 2013 with a clinical diagnosis of malaria were examined for the presence of asexual forms of P. falciparum in peripheral blood smears. All smear positive febrile patients who met the eligibility criteria were enrolled. Smear negative fever cases were enrolled as control cases. RDTs were performed using both urine and blood samples by using commercially available blood specific kits. Results: Sixty blood smear positive cases and 51 febrile blood smear negative cases were enrolled. Sensitivity and specificity of RDT urine were 86.67% (95%CI:75.83–93.09) and 94.12% (95%CI:84.08–97.98) respectively whereas those of RDT blood were 91.67% (95% CI: 81.93–96.39) and 98.04% (95% CI 89.7–99.65). The sensitivity of both RDT urine as well as RDT blood were found to be dependent on the level of parasitemia. Conclusion: Results of this study are promising. Larger studies are needed to assess whether RDTs using urine could serve as a practical, reliable method for the detection of P. falciparum in a non-invasive manner where invasive blood taking is less feasible.

The present study is done to study different aspects of Helicobacter pylori (H. pylori) such as its prevalence, association with upper gastrointestinal pathology, diagnosis and treatment outcome. Gastric antral biopsy and serology for H. pylori was done for all dyspeptic patients. Histopathology, gram stain and biopsy urease test was done from the gastric biopsy specimen. The prevalence of H. pylori infection was 58.8%. The sensitivity, specificity, positive and negative predictive value for histopathology was 96.9%, 100%, 100% and 95.8%, respectively; for biopsy urease test 80.4%, 100%, 100% and 78.2%, respectively; for gram stain 85.6%, 97.1%, 97.6% and 82.5%, respectively, and for serology 94.8%, 77.9%, 86% and 91.4%, respectively. Mostly peptic ulcer and duodenitis cases followed by chronic active gastritis were associated with H. pylori infection. Repeat biopsy revealed eradication of H. pylori in 90.7% cases. In dyspeptic patients, endoscopic biopsy not only detects H. pylori infection, but also reveals different gastric pathologies.

Plasmodial lactate dehydrogenase enzyme is known to be a potential immunodiagnostic marker as well as a novel target for chemotherapy and hence its economical bulk production is required. Although Escherichia coli offers a mean for rapid, high yield, and economical production of recombinant proteins, high-level production of functional recombinant Plasmodium falciparum lactate dehydrogenase enzyme in E. coli is quite challenging. To explore a new approach for high level soluble expression of PfLDH in E. coil, the recombinant clone carrying the transgene was subjected to optimizations of expression conditions at shake flask level. The comparative analysis of different culture parameters revealed that modified-TB medium at a specific pH has the highest capacity to produce the high level of expression of PfLDH. There was an increase of 23% pure protein from the culture grown under optimised conditions in comparison to normal medium. The purified rPfLDH was found to be biologically active with specific activity of 485.3 mu mol/min/mg. We believe that this strategy could be of special interest due to its capacity to improve the expression level of PfLDH and the procedure described in this study may provide a reliable and simple method for production of large quantities of soluble and biologically active PfLDH.

The C-terminal portion of Plasmodium falciparum Merozoite Surface Protein1(42)kDa known to have prophylactic potential was expressed using Pichia pastoris. Bioinformatics based B-cell and T-cell epitope predictions revealed uniform distribution of epitopes on MSP1(42)kDa protein. In order to have a broad spectrum protection with multiple epitopes, the complete MSP1(42)kDa was selected for expression using P. pastoris. The gene fragment encoding the MSP1(42)kDa was amplified from the P falciparum genomic DNA and cloned into pPIC9K yeast transfer vector under the control of AOX1 promoter in fusion with the alpha secretory signal. The expression cassette was integrated into the Pichia genome via homologous recombination. Recombinant Pichia clones carrying multi copy integrants of the transgene were selected based on geneticin tolerance. The positive Pichia clone was methanol induced to express the transgene and the expression of the recombinant MSP1(42)kDa was confirmed via immunoblotting and MALDI-TOF analysis. Purification strategy were developed and verified by SDS-PAGE electrophoresis. Immunization of rabbit with recombinant MSP1(42)kDa in Freund's adjuvant resulted in high antibody titers against the recombinant MSP-1(42)kDa. Rabbit produced anti-MSP1IgG were inhibited in vitro parasite growth, demonstrating that this inhibition anti-MSP1IgG mediated. The multicopy recombinant Pichia transformant obtained in the present study has an immense industrial application for large scale production of MSP1(42)kDa recombinant protein for either prophylactic or diagnostic applications.

Cerebral malaria is a severe neuropathological complication of Plasmodium falciparum infection. It results in high mortality and post-recovery neuro-cognitive disorders in children, even after appropriate treatment with effective anti-parasitic drugs. While the complete landscape of the pathogenesis of cerebral malaria still remains to be elucidated, numerous innovative approaches have been developed in recent years in order to improve the early detection of this neurological syndrome and, subsequently, the clinical care of affected patients. In this review, we briefly summarize the current understanding of cerebral malaria pathogenesis, compile the array of new biomarkers and tools available for diagnosis and research, and describe the emerging therapeutic approaches to tackle this pathology effectively.

The safe disposal and utilisation of coal fly ash (CFA) for value addition are still major problems worldwide. Al2O3 is a major valued material associated with it. Till date no methods are available to treat CFA for recovering its valued materials. Leaching of alumina from CFA in an economical way is very difficult, which either requires higher chemical consumption or application of stringent reaction conditions such as high temperature and pressure. In this paper a simple alternative method has been attempted to dissolve alumina from CFA. Addition of fluoride ion as HF improved the acid leaching performance of fly ash to a large extent. XRD data showed mullite, the alumina bearing phase, as the major phase which gets dissolved during leaching operation. A standard procedure for the precipitation of alumina can be employed after the leached solution is obtained.