December 2020 – LeishDB (protozoa), Universidad Mayor

Membrane dynamics in Leishmania amazonensis and antileishmanial activities of β-carboline derivatives

December 10, 2020 at 9:21 am (last edited December 15, 2020 at 3:09 pm)

Two β-carboline compounds, 8i and 6d, demonstrated in vitro antileishmanial exercise towards Leishmania (L.) amazonensis promastigotes just like that of miltefosine (MIL). Estimates of the membrane-water partition coefficient (OkM/W) and the compound concentrations within the membrane (cm50) and aqueous section (cw50) for half maximal inhibitory focus had been made. Whereas these biophysical parameters for 6d weren’t considerably completely different from these reported for MIL, 8i confirmed decrease affinity for the parasite membrane (decrease OkM/W) and a decrease focus of the compound within the membrane required to inhibit the expansion of the parasite (decrease cm50). A 2-hour remedy of Leishmania promastigotes with the compounds 8i and 6d prompted membrane rigidity in a concentration-dependent method, as demonstrated by the electron paramagnetic resonance (EPR) method and spin label methodology. This elevated rigidity of the membrane was interpreted to be related to the incidence of cross-linking of oxidized cytoplasmic proteins to the parasite membrane skeleton. Importantly, the 2 β-carboline-oxazoline derivatives confirmed low hemolytic motion, each in experiments with remoted purple blood cells or with complete blood, denoting their nice Leishmania/erythrocyte selectivity index. Utilizing electron microscopy, modifications within the membrane of each the amastigote and promastigote type of the parasite had been confirmed, and it was demonstrated that compounds 8i and 6d decreased the variety of amastigotes in contaminated murine macrophages. Moreover, 8i and 6d had been extra poisonous to the protozoa than to J774A.1 macrophages, with handled promastigotes exhibiting a lower in cell quantity, mitochondrial membrane potential depolarization, accumulation of lipid our bodies, elevated ROS manufacturing and modifications within the cell cycle. Serum samples had been collected from 250 canine and examined utilizing oblique enzyme-linked immunosorbent assay (ELISA) and oblique immunofluorescence antibody exams (IFATs). Information regarding canine, their surroundings, and their house owners’ data of leishmaniasis had been collected utilizing a questionnaire. To find out RFs for contact with the parasite, univariate statistical evaluation based mostly on chi-squared and Fisher’s actual exams, adopted by logistic regression, was used. Canine visceral leishmaniasis in space with current Leishmania transmission: prevalence, prognosis, and molecular identification of the infecting species Introduction: Canine visceral leishmaniasis (CVL) is an endemic illness in Brazil, and built-in management actions have been adopted by the Brazilian Ministry of Well being to manage its unfold. Nevertheless, the transmission profile is unknown in areas with current CVL instances, together with Itaúna, positioned within the Brazilian state of Minas Gerais, the place the current examine was carried out. Strategies: A complete of two,302 canine from 12 neighborhoods had been serologically examined for canine VL utilizing the present diagnostic protocol adopted by the Brazilian Ministry of Well being. Check positivity fee (TPR) and CVL prevalence had been decided for every neighborhood. The presence of Leishmania was assessed in 60 seropositive canine which had been really helpful for euthanasia. Twenty-two of them (37%) had been asymptomatic, and 38 (63%) had been symptomatic for CVL. Parasitological (myeloculture and smear/imprint) and molecular (PCR) strategies had been employed for Leishmania detection in bone marrow, spleen, mesenteric lymph nodes, and ear […]

Expanded genome-wide comparisons give novel insights into population structure and genetic heterogeneity of Leishmania tropica complex

76 Mitchell

December 10, 2020 at 9:21 am (last edited December 12, 2020 at 4:51 pm)

Leishmania tropica is likely one of the essential causative brokers of cutaneous leishmaniasis (CL). Inhabitants constructions of L. tropica look like genetically extremely numerous. Nonetheless, the connection between L. tropica strains genomic variety, protein coding gene evolution and biogeography are nonetheless poorly understood. On this examine, we sequenced the genomes of three new medical L. tropica isolates, two derived from a current outbreak of CL in camps internet hosting Syrian refugees in Lebanon and one historic isolate from Azerbaijan to additional refine comparative genome analyses. In silico multilocus microsatellite typing (MLMT) was carried out to combine the present variety of genome sequence knowledge within the wider accessible MLMT genetic inhabitants framework. Single nucleotide polymorphism (SNPs), gene copy quantity variations (CNVs) and chromosome ploidy had been investigated throughout the accessible 18 L. tropica genomes with a essential deal with protein coding genes. MLMT divided the strains in three populations that broadly correlated with their geographical distribution however not populations outlined by SNPs. Distinctive SNPs profiles divided the 18 strains into 5 populations primarily based on principal element evaluation. Gene ontology enrichment evaluation of the protein coding genes with inhabitants particular SNPs profiles revealed varied organic processes, together with iron acquisition, sterols synthesis and drug resistance. This examine additional highlights the complicated hyperlinks between L. tropica essential genomic heterogeneity and the parasite broad geographic distribution. Distinctive sequence options in protein coding genes recognized in distinct populations reveal potential novel markers that could possibly be exploited for the event of extra correct typing schemes to additional enhance our data of the evolution and epidemiology of the parasite in addition to highlighting protein variants of potential practical significance underlying L. tropica particular biology. First report of an Onchocercidae worm infecting Psychodopygus carrerai carrerai sandfly, a putative vector of Leishmania braziliensis within the Amazon Sandflies are bugs of public well being curiosity on account of their function as vectors of parasites of the genus Leishmania, in addition to different pathogens. Psychodopygus carrerai carrerai is taken into account an essential sylvatic vector of Leishmania (Viannia) braziliensis in Amazonia. On this examine, sandflies had been collected in a forested space within the Xapuri municipality, within the State of Acre (Northern Brazil). Two Ps. carrerai carrerai females had been discovered parasitized with a larval type of a filarial worm, one within the labium of the proboscis, the opposite after the top was squashed, suggesting they had been infective larvae. Sandflies had been recognized by means of morphological characters in addition to amplification and sequencing of the cytochrome oxidase gene (COI). This was the primary sequence obtained for Ps. carrerai carrerai for this marker. The obtained nematodes had been additionally characterised by means of direct sequencing of a fraction of COI and 12S genes, each mitochondrial, and ITS1, a nuclear marker. Phylogenetic analyses revealed that the filarial nematodes belong to a species with out sequences for these markers within the database, a part of household Onchocercidade and carefully associated to genus Onchocerca (12S tree). Though sandfly an infection with […]

An overview of the sandfly fauna (Diptera: Psychodidae) followed by the detection of Leishmania DNA and blood meal identification in the state of Acre, Amazonian Brazil

75 Mitchell

December 10, 2020 at 9:21 am (last edited December 12, 2020 at 4:52 pm)

Background: In Acre state, Brazil, the dissemination of cutaneous leishmaniasis has elevated in recent times, with restricted information of the potential Leishmania spp. vectors concerned. Goals: Right here, information in regards to the sandfly fauna of Brasiléia municipality, Leishmania DNA-detection charges and the identification of blood meal sources of bugs captured in 2013-2015 are introduced. Strategies: Parasite detection in feminine sandflies was carried out individually by multiplex polymerase chain response (PCR) (Leishmania kDNA/sandfly cacophony-gene), with the identification of Leishmania spp. by hsp70-PCR and sequencing. The identification of blood gut-content from fed females was carried out by cyt b-PCR and sequencing. Findings: A complete of 4,473 sandflies have been captured. A subgroup of 864 non-blood-fed females evaluated for the presence of Leishmania DNA confirmed 2.9% positivity for Leishmania (Viannia) braziliensis and L. (V.) guyanensis. The identification of blood meal sources was carried out in 96 blood-fed females, permitting the identification of 13 vertebrate species. In 9/96 fed females, DNA from L. (V.) shawi, L. (V.) guyanensis, L. (V.) braziliensis and Endotrypanum sp. was detected. Most important conclusions: In Brumptomyia sp. and Evandromyia termitophila, the primary report of Leishmania DNA-detection is offered in Acre; Nyssomyia shawi is implicated as potential vector of L. (V.) braziliensis and L. (V.) guyanensis for the primary time in Brazil. Parasites of Leishmania genus have developed refined methods permitting them to deactivate their host macrophage to advertise their survival. It has develop into clear that miRNAs play vital roles in shaping innate and adaptive immune responses towards pathogens. It’s not stunning that a number of pathogens together with Leishmania have developed the flexibility to control host macrophage miRNA expression so as to manipulate host cell phenotypes to their benefit. Nevertheless, little or no is understood in regards to the mechanisms utilized by intracellular pathogens to drive adjustments in host cell miRNA abundance. On this overview, Leishmania exploitation of macrophage transcription issue c-Myc as a vital proxy virulence issue to control abundance of macrophage miRNAs influencing macrophage physiology to advertise its survival might be mentioned Metformin promotes susceptibility to experimental Leishmania braziliensis an infection Background: Metformin (MET) is a hypoglycemic drug used for the remedy of diabetes, regardless of interference in host immunity in opposition to microorganisms. Cutaneous an infection brought on by pathogens equivalent to Leishmania braziliensis (Lb), the agent liable for cutaneous leishmaniasis (CL) in Brazil, represents an attention-grabbing mannequin during which to guage the consequences related to MET. Goal: To guage the modulatory impact of MET in Lb an infection. Materials and strategies: Experimental research of Lb an infection and MET remedy in BALB/c mice and Uncooked 264.7 macrophages. Findings: MET remedy interfered with lesion kinetics, elevated parasite load and decreased macrophage proliferation. Low concentrations of MET in Lb tradition permit for the upkeep of stationary parasite development section. Lb-infected cells handled with MET exhibited elevated parasite load. Whereas each MET and Lb an infection alone promoted the manufacturing of intracellular reactive oxygen species (ROS), decreased ranges of ROS have been seen in MET-treated Lb-infected macrophages. Most important conclusion: Experimental remedy with MET interfered with the kinetics […]

Genomic and Transcriptomic Analysis for Identification of Genes and Interlinked Pathways Mediating Artemisinin Resistance in Leishmania donovani

72 Mitchell

December 10, 2020 at 9:21 am

Current treatment for visceral leishmaniasis (VL), compromised by drug resistance, toxicity, and extreme worth, requires for less complicated, safer, and low-cost medication. Artemisinin has been found to be an effectual drug varied in experimental fashions of leishmaniasis. Comparative genome and transcriptome analysis of in vitro-adapted artesunate-resistant and -sensitive wild-type Leishmania donovani parasites was carried out using next-generation sequencing and single-color DNA microarray know-how, respectively, to determine genes and interlinked pathways contributing to drug resistance. Full-genome sequence and, 237 insertion deletions, 616 copy amount variations (CNVs) (377 deletions and 239 duplications), and trisomy of chromosome 12 in Okay133AS-R parasites. Transcriptome analysis revealed differential expression of 208 genes (fold change ≥ 2) in Okay133AS-R parasites. Purposeful categorization and analysis of modulated genes of interlinked pathways recognized plausible permutations in Okay133AS-R parasites, paying homage to (i) a dependency on lipid and amino acid metabolism for producing energy, (ii) diminished DNA and protein synthesis leading to parasites throughout the quiescence state, and (iii) vigorous drug efflux. The upregulated expression of cathepsin-L like protease, amastin-like ground protein, and amino acid transporter and downregulated expression of the gene encoding ABCG2, pteridine receptor, adenylatecyclase-type receptor, phosphoaceylglucosamine mutase, and positive hypothetical proteins are concordant with genomic alterations suggesting their potential place in drug resistance. The look at equipped an understanding of the molecular basis linked to artemisinin resistance in Leishmania parasites, which will be advantageous for safeguarding this drug for future use. Polyvinyl alcohol-based electrospun matrix as a provide system for nanoemulsion containing chalcone in direction of Leishmania (Leishmania) amazonensis Cutaneous leishmaniasis is a worldwide public nicely being draw back. Typical therapies, together with the extreme worth, have many hostile outcomes and circumstances of parasite’s resistance. Chalcones are secondary metabolites precursors throughout the flavonoid pathway and is perhaps obtained naturally, nevertheless with low yield from plant raw supplies. Thus, the utilization of synthetic chalcones has been a promising approach for the occasion of molecules with leishmanicidal train. Thus, this work aimed to develop a managed launch system of two synthetic chalcone (trans-chalcones and three’-(trifluormethyl)-chalcone) using polyvinyl alcohol nanofibers (PVA) as scaffold. The affiliation of chalcones to the nanofibers was made by nanoemulsions (NE) thereof, i.e., a colloidal system on a nanometric scale, which allows compounds with reverse polarities to remain miscible and regular all by their manipulation. Chalcone nanoemulsions had been developed using the spontaneous emulsification method. The NE had been characterised referring to their particle measurement, polydispersion index (PDI), and zeta potential. The outcomes confirmed NE with spherical kind, absolute values of zeta potential had been larger than 30 mV and homogeneous distribution pattern (PDI < 0.3). Dynamics mild scattering (DLS) analysis confirmed comparable hydrodynamic rays, i.e., 180 nm (trans-chalcone NE) and 178 nm (NE containing 3′-(trifluormethyl)-chalcone, together with presenting encapsulation effectivity values close to 100 %. Subsequently, the NE had been added to a polymeric reply of polyvinyl alcohol (PVA) and processed by the electrospinning method affording a PVA matrix (15 %, w/v) nanofiber containing the chalcones NE at 1 mg.mL-1. In a follow-up experiment, the pores […]

Large-scale cultivation of Leishmania infantum promastigotes in stirred bioreactor

70 Mitchell

December 10, 2020 at 9:21 am

Background & goals: Bioreactors are sensible instruments which are used for economical, time-conserving and large-scale manufacturing of biomass from cell cultivation. They supply optimum environmental situations similar to pH and temperature required for acquiring most quantities of biomass. Nonetheless, there is no such thing as a proof within the literature on the large-scale cultivation of Leishmania infantum parasites within the bioreactor. Due to this fact, the current research was undertaken to develop a brand new strategy for acquiring L. infantum biomass by utilizing pH and temperature controllable stirred bioreactor and to match parasitic development kinetics with classical technique inside erlenmeyers. Strategies: With a purpose to receive parasite biomass, a newly developed pH and temperature managed stirred bioreactor was used and its efficacy was in contrast with a graduated classical scale-up technique. Progress kinetics of parasites inside erlenmeyers and bioreactors have been decided by evaluating promastigote numbers utilizing haemocytometer. The graduated scale enlargement of tradition was adopted by T25 flask, T75 flask, and 1 L erlenmeyer, respectively. Outcomes: Obtained outcomes confirmed a 10-fold improve within the variety of promastigotes throughout the typical tradition carried out in 700 ml medium, whereas parasite numbers elevated roughly 15 occasions as a consequence of preliminary inoculation quantities within the bioreactor tradition carried out within the 3.5 l medium. Thus, there was 7.5 occasions extra biomass assortment in bioreactor in comparison with classical technique. Interpretation & conclusion: It’s postulated that fixed tradition pH and temperature within the bioreactor extends cultivation time. This might result in important improve in parasite numbers. Therefore, pH and temperature controllable bioreactors supplied acquisition of enough quantities of biomass in distinction to classical strategies. Due to this fact, this kind of bioreactors could substitute classical tradition strategies within the manufacturing of antigenic molecules for vaccine improvement. Investigating associations between intestinal alterations and parasite load in response to Bifidobacterium spp. and Lactobacillus spp. abundance within the intestine microbiota of hamsters contaminated by Leishmania infantum Background: Visceral leishmaniasis (VL) is a tropical uncared for illness with excessive related charges of mortality. A number of research have highlighted the significance of the intestinal tract (IT) and intestine microbiota (GM) within the host immunological protection. Knowledge within the literature on parasite life cycle and host immune protection towards VL are scarce concerning the results of an infection on the IT and GM. Aims: This research aimed to research modifications noticed within the colon of Leishmania infantum-infected hamsters, together with alterations within the enteric nervous system (ENS) and GM (particularly, ranges of bifidobacteria and lactobacilli). Strategies: Male hamsters have been inoculated with L. infantum and euthanised at 4 or eight months post-infection. Intestines have been processed for histological evaluation and GM evaluation. Quantitative polymerase chain response (qPCR) was carried out to quantify every group of micro organism: Bifidobacterium spp. (Bf) and Lactobacillus spp (LacB). Findings: Contaminated hamsters confirmed histoarchitectural loss within the colon wall, with elevated thickness within the submucosa and the mucosa layer, in addition to higher numbers of intraepithelial lymphocytes. Varieties suggestive of amastigotes have been seen inside mononuclear cells. L. infantum an infection induced […]

The potential therapeutic role of PTR1 gene in non-healing anthroponotic cutaneous leishmaniasis due to Leishmania tropica

69 Mitchell

December 10, 2020 at 9:20 am (last edited December 21, 2020 at 1:42 pm)

Background: Drug resistance is a typical phenomenon ceaselessly noticed in nations the place leishmaniasis is endemic. Because of the manufacturing of the pteridine reductase enzyme (PTR1), medicine lose their efficacy, and consequently, the affected person turns into unresponsive to therapy. This research aimed to check the in vitro impact of meglumine antimoniate (MA) on non- therapeutic Leishmania tropica isolates and on MA transfected non-healing one to PTR1. Strategies: Two non-healing and one therapeutic isolates of L. tropica have been collected from sufferers who obtained two programs or one cycle of intralesional MA together with biweekly liquid nitrogen cryotherapy or systemic therapy alone, respectively. After affirmation of L. tropica isolates by polymerase chain response (PCR), the recombinant plasmid pcDNA-rPTR (antisense) was transfected by way of electroporation and cultured on M199. Isolates in type of promastigotes have been handled with totally different concentrations of MA and skim utilizing an enzyme-linked immunosorbent assay (ELISA) reader and the half inhibitory focus (IC50 ) worth was calculated. The amastigotes have been grown in mouse macrophages and have been equally handled with varied concentrations of MA. The tradition glass slides have been stained, and the imply variety of intramacrophage amastigotes and contaminated macrophages have been assessed in triplicate for each levels. Outcomes: All three transfected isolates displayed a discount in optical density in contrast with the promastigotes in respective isolates, though there was no vital distinction between non-healing and therapeutic isolates. In distinction, within the medical kind (amastigotes), there was a major distinction between non-healing and therapeutic isolates (p < 0.05). Conclusion: The outcomes indicated that the PTR1 gene diminished the efficacy of the drug, and its inhibition by antisense and will enhance the therapy of non-healing circumstances. These findings have future implications within the prophylactic and therapeutic modality of non- therapeutic Leishmania isolates to drug. Leishmanicidal, cytotoxic and apoptotic results of Gossypium hirsutum bulb extract and its separated fractions on Leishmania main Background & targets: Leishmaniasis is a serious world well being downside with no protected and efficient therapeutic medicine. This research evaluated the cytotoxic and apoptotic results of crude extract and fractions of Gossypium hirsutum bulb on Leishmania main levels utilizing superior experimental fashions. Strategies: Bulbs of G. hirsutum have been collected from the Kerman province of Iran. The bulb was extracted utilizing Soxhlet equipment and totally different fractions have been obtained by column chromatography (CC). Completely different concentrations of the extract and the fractions have been evaluated in opposition to L. main and in contrast with Glucantime®. The cytotoxicity and apoptotic values have been analysed by circulation cytometry. The fractions obtained in CC have been monitored by skinny layer chromatography, and fractions with comparable chromatographic patterns have been blended. Outcomes: The extract and two fractions, F4 and F5 inhibited the proliferation of L. main promastigotes and amastigotes in a dose-dependent method at 72 h post-treatment. No vital cytotoxic results have been noticed for extract and fractions, because the selectivity index was over 1000, far past >10. The imply apoptotic values for L. main have been superior to these of Glucantime. Interpretation & conclusion: Each the crude […]