The ParF protein of plasmid TP228 belongs to the ubiquitous superfamily ParA ATPases that drive DNA segregation in bacteria. ATP-bound polymerizes into multistranded filaments. partner ParG is dimeric, consisting C-termini interweave a ribbon–helix–helix domain contacting centromeric and unstructured N-termini. stimulates ATP hydrolysis by ≈30-fold. Here, we establish mobile tails are crucial for this enhancement arginine R19 within tail absolutely required activation nucleotide hydrolysis....

Leishmania spp. is a protozoan parasite that affects millions of people around the world. At present, there no effective vaccine to prevent leishmaniases in humans. A major limitation development lack precise understanding particular immunological mechanisms allow survival host. The parasite-host cell interaction induces dramatic changes transcriptome patterns both organisms, therefore, detailed analysis gene expression infected tissues will contribute evaluation drug and candidates,...

Background Patients with chronic Chagas disease present marked clinical and immunological heterogeneity. During the disease, multiple immune mechanisms are activated to fight parasite. The purpose of this study was investigate expression patterns genes involved in relevant processes throughout patients disease. Methodology/Principal findings High-throughput RT-qPCR QuantStudio 12K Flex real-time PCR system used evaluate 106 immune-related PBMC from a cohort cardiac (CCC I), asymptomatic...

Abstract Purpose Fever of intermediate duration (FID) is defined as a fever in the community without specific origin or focus, with between 7 and 28 days. FID often caused by pathogens associated animal contact their arthropods parasites, such ticks, fleas, lice. The purpose this work to design collection molecular tools promptly accurately detect common bacterial causing FID, including bacteria belonging genera Rickettsia , Bartonella Anaplasma Ehrlichia well Coxiella burnetii. Methods...

Summary The high variability among strains and isolates of Trypanosoma cruzi the existence shared antigenic determinants with other pathogens, particularly members Leishmania genus make difficult specific diagnosis Chagas' disease. data reported in this paper show that T. KMP11 protein is an immunodominant antigen highly recognized by sera from chagasic leishmaniasis patients. By use amino- carboxyl-terminal truncated recombinant proteins synthetic peptides, evidence provided while patients...

ParG is the prototype of a group small (<10 kDa) proteins involved in accurate plasmid segregation. The protein dimeric DNA binding factor, which consists symmetric paired C-terminal domains that interleave into ribbon-helix-helix fold crucial for interaction with DNA, and unstructured N-terminal previously unknown function. Here shown to be transcriptional repressor parFG genes. assembles on its operator site initially as tetramer (dimer dimers) and, at elevated concentrations, pair...

Background: Acute respiratory infections are one of the major causes morbidity and mortality in children under 5 years developing countries a challenge for health system these countries. In Cabo Verde, despite lack recent studies, data indicate that it affects thousands children, being fourth leading cause infant 2013. The aim this study was to identify describe etiological agents associated with acute tract old, their risk factors, such as clinical symptoms or socio-demographic...

The interaction of Leishmania with BALB/c mice induces dramatic changes in transcriptome patterns the parasite, but also target organs (spleen, liver…) due to its response against infection. Real-time quantitative PCR (qPCR) is an interesting approach analyze these and understand immunological pathways that lead protection or progression disease. However, qPCR results need be normalized one more reference genes (RG) correct for non-specific experimental variation. development technical...

Chagas disease, caused by the

This prospective study aims to diagnose the etiology of non-focalized fever lasting between 5 and 28 days in islands La Palma El Hierro (Canary Islands, Spain) during 2021, using serology PCR.

A cDNA clone codifying ribosomal protein L25 was isolated from a Leishmania braziliensis gene library. The alignment of the amino acid sequence deduced this with other proteins revealed that is related to L23/25 family. This so because shows, in its C-terminal end, rRNA binding domains characteristic these and at N-terminal end NLS necessary for entry into nucleus. Southern blot analysis showed 2 copies per genome arranged tandem position pointing same direction. Northern transcribed mRNAs...

Cross-reactions between Leishmania braziliensis and Trypanosoma cruzi caused by common antigenic determinants hinder the specific diagnosis of cutaneous mucocutaneous leishmaniasis (CL MCL). Therefore, usefulness 70-kDa heat shock protein (Hsp70) from L. for differential serodiagnosis was investigated. The single-copy gene encoding Hsp70, consisting 663 amino acids, isolated a genomic DNA library. antigenicity data show that Hsp70 is an immunodominant antigen highly recognized (84%) sera...

Infection by the Trypanosoma cruzi parasite causes Chagas disease and triggers multiple immune mechanisms in host to combat pathogen. has a variable clinical presentation progression, producing chronic phase fragile balance between response replication that keeps patients clinically silent asymptomatic stage for years. Since is intracellular replicates within cells, cell-mediated of adaptive immunity plays critical role. This function mainly orchestrated T lymphocytes, which recognize...

The humoral immune response against Leishmania braziliensis histone H1 by patients with cutaneous leishmaniasis is described. For this purpose, the protein was purified as a recombinant in prokaryotic expression system and assayed enzyme-linked immunosorbent assay (ELISA) collection of sera from Chagas' disease. assays showed that L. recognized 66% serum samples 40% disease, indicating it acts an immunogen during leishmaniasis. In order to locate linear antigenic determinants protein,...

Transcriptional analysis is a useful approximation towards the identification of global changes in host-pathogen interaction, order to elucidate tissue-specific immune responses that drive immunopathology disease. For this purpose, expression 223 genes involved innate and adaptive response, lipid metabolism, prostaglandin synthesis, C-type lectin receptors MAPK signaling pathway, among other processes, were analyzed during early infection spleens BALB/c mice infected by Leishmania infantum....

Leishmania infection of phagocytic cells, such as macrophages, induces the differentiation infected cells into different phenotypes according to their surrounding microenvironments. The classical activation macrophages involves metabolic reprogramming, in which several metabolites succinate, fumarate and itaconate are accumulated. immunoregulatory functions context were investigated this paper. Ex vivo bone marrow-derived differentiated classically activated through IFNG with infantum. A...

The isolation and molecular characterization of the histone H1-encoding gene from Leishmania braziliensis was carried out. is present in genome as a single copy transcribed polyadenylated transcript 830 nucleotides. deduced amino acid sequence has its central region DNA binding K-[K/R]-A-A-[A/P] motif, which repeated tandem 9 times.

Aims: To develop a rapid colourimetric assay for the detection of Toxoplasma gondii DNA using polystyrene beads as solid support. Methods and Results: A nested‐polymerase chain reaction (PCR)‐ELISA T. was standardized by optimizing hybridization time probe concentration. Its threshold then determined compared with Southern blotting hybridization. These were found to be equivalent, but PCR‐ELISA‐beads test is easier perform turnaround much shorter than blot. Conclusions: The valuable tool...

The isolation and molecular characterization of the gene coding for L14 ribosomal protein from L. braziliensis is described. There are 2 copies per haploid genome, repeated in a head-to-tail tandem orientation located single chromosome approximately 950 kb. Northern blot analyses indicate presence transcript 0·95 kb which up-regulated when parasites reach stationary growth phase. codes 175 amino acid long polypeptide showing 75–83% sequence identity with proteins trypanosomatids 25% its...

Transcriptional analysis of complex biological scenarios has been used extensively, even though sometimes the results such may prove imprecise or difficult to interpret due an overwhelming amount information. In this study, a large-scale real-time qPCR experiment was coupled multivariate statistical in order describe main immunological events underlying early L. infantum infection livers BALB/c mice. High-throughput evaluate expression 223 genes related response and metabolism 1, 3, 5, 10...

<title>Abstract</title> The present prospective study aims to diagnose, through serology and PCR, the etiology of non-focalized fever lasting between 5 28 days in islands La Palma El Hierro (Canary Islands, Spain) during year 2021. etiological profile described this is similar that studies intermediate duration (FDI), with zoonoses being main cause. Murine typhus (MT) identified as primary cause, followed by Q (QF). incidence MT highest published nationally Europe, 39.6 cases 79.7 Hierro....

In this paper, we report a method for isolation of high molecular weight DNA from Leishmania promastigotes. This technique is especially indicated small-scale purification suitable the construction highly representative genomic libraries. our protocol, lysis buffer compatible with RNase treatment, avoiding an additional precipitation step and consequent shearing DNA. order to prove quality isolated by method, braziliensis library was constructed, L. KMP-11 gene cloned after screening...

Histone H1 in Leishmania presents relevant differences compared to higher eukaryote counterparts, such as the lack of a DNA-binding central globular domain. Despite that, it is apparently fully functional since its differential expression levels have been related changes chromatin condensation and infectivity, among other features. The localization aggregation state L. braziliensis has determined by immunolocalization, mass spectrometry, cross-linking electrophoretic mobility shift assays....

By screening a Leishmania braziliensis complementary DNA library with pool of sera from leishmaniasis patients, the gene coding for L6 ribosomal protein was isolated. The sequence, genomic organization, and transcription this are described in article. sequence analysis L. shows single open reading frame, which codes 192 amino acids (aa) hypothetical molecular mass 20.9 kDa. exhibits significant similarity to proteins higher eukaryotes yeast. Thus, contains 4 functional motifs, located at...

Sera from individuals suffering leishmaniasis have been shown to strongly react against conserved proteins the parasite, such as ribosomal, histones and heat-shock proteins. Some of these also described immunogenic in several auto-immune syndromes, detection antibodies them is considered be indicative disorder immune system. In this paper, we investigate whether there any relationship between recognition some Leishmania braziliensis by cutaneous (CL) mucocutaneous (MCL) leishmaniasis, human...