Genome Sequence of the Native Apiculate Wine Yeast Hanseniaspora vineae T02/19AF

The use of novel yeast strains for winemaking improves quality and provides variety including subtle characteristic differences in fine wines. Here we report the first genome of a yeast strain native to Uruguay, Hanseniaspora vineae T02/19AF, which has been shown to positively contribute to aroma and wine quality.Fil: Giorello, Facundo M.. Universidad de la República; UruguayFil: Berná, Luisa. Instituto Pasteur de Montevideo; UruguayFil: Greif, Gonzalo. Instituto Pasteur de Montevideo; UruguayFil: Camesasca, Laura. Inst. de Investigaciones Biológicas Clemente Estable; UruguayFil: Salzman, Valentina. Instituto Pasteur de Montevideo; Uruguay. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Medina, Karina. Universidad de la Republica. Facultad de Química; UruguayFil: Robello, Carlos. Instituto Pasteur de Montevideo; UruguayFil: Gaggero, Carina. Inst. de Investigaciones Biológicas Clemente Estable; UruguayFil: Aguilar, Pablo S.. Instituto Pasteur de Montevideo; UruguayFil: Carrau, Francisco. Sección Enología; Urugua

TcI isolates of Trypanosoma cruzi exploit the antioxidant network for enhanced intracellular survival in macrophages and virulence in mice

Trypanosoma cruzi species is categorized into six discrete typing units (TcI to TcVI) of which TcI is most abundantly noted in the sylvatic transmission cycle and considered the major cause of human disease. In our study, the TcI strains Colombiana (COL), SylvioX10/4 (SYL), and a cultured clone (TCC) exhibited different biological behavior in a murine model, ranging from high parasitemia and symptomatic cardiomyopathy (SYL), mild parasitemia and high tissue tropism (COL), to no pathogenicity (TCC). Proteomic profiling of the insect (epimastigote) and infective (trypomastigote) forms by two-dimensional gel electrophoresis/ matrix-assisted laser desorption ionization-time of flight mass spectrometry, followed by functional annotation of the differential proteome data sets (≥2-fold change, P<0.05), showed that several proteins involved in (i) cytoskeletal assembly and remodeling, essential for flagellar wave frequency and amplitude and forward motility of the parasite, and (ii) the parasite-specific antioxidant network were enhanced in COL and SYL (versus TCC) trypomastigotes. Western blotting confirmed the enhanced protein levels of cytosolic and mitochondrial tryparedoxin peroxidases and their substrate (tryparedoxin) and iron superoxide dismutase in COL and SYL (versus TCC) trypomastigotes. Further, COL and SYL (but not TCC) were resistant to exogenous treatment with stable oxidants (H2O2 and peroxynitrite [ONOO-]) and dampened the intracellular superoxide and nitric oxide response in macrophages, and thus these isolates escaped from macrophages. Our findings suggest that protein expression conducive to increase in motility and control of macrophage-derived free radicals provides survival and persistence benefits to TcI isolates of T. cruzi.Fil: Zago, María Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; ArgentinaFil: Hosakote, Yashoda M.. University of Texas Medical Branch; Estados UnidosFil: Koo, Sue Jie. University of Texas Medical Branch; Estados UnidosFil: Dhiman, Monisha. University of Texas Medical Branch; Estados UnidosFil: Piñeyro, María Dolores. Instituto Pasteur de Montevideo. Unidad de Biología Molecular; Uruguay. Universidad de la Republica; UruguayFil: Parodi­ Talice, Adriana. Instituto Pasteur de Montevideo. Unidad de Biología Molecular; Uruguay. Universidad de la Republica; UruguayFil: Basombrío, Miguel Ángel Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; ArgentinaFil: Robello, Carlos. Instituto Pasteur de Montevideo. Unidad de Biología Molecular; Uruguay. Universidad de la Republica; UruguayFil: Garg, Nisha J.. University of Texas Medical Branch; Estados Unido

Genomic Organization of Trypanosoma cruzi tRNA Genes

A frequently overlooked category of genes in large-scale genome analyses is the nonprotein-coding genes. Specifically, the genomic organization of tRNA genes in Trypanosoma cruzi—a unicellular eukaryotic pathogen responsible for causing the debilitating human disease Chagas disease—has largely remained uncharted due to the previous incompleteness of the genome assembly. By utilizing advanced genomic techniques and improved assembly methods, the quality of genome assemblies has significantly improved, making these studies feasible. Here, we analyzed the genic content and distribution of tRNA genes in the nuclear genome of various strains compared with the genome organization of other related trypanosomatids. We found synteny in most of the tDNAs clusters between T. cruzi and Trypanosoma brucei, highlighting the significance of the genomic location of these Pol III-transcribed genes. A vast majority of the isoacceptor species are encoded by two genes, except for tDNASeC, which comprises a tandem of 11 copies in the core compartment associated with well-positioned nucleosomes. Finally, we describe a group of tRNA genes located at chromatin folding domain boundaries, potentially acting as chromatin insulators in T. cruzi.Agencia Nacional de Investigación e InnovaciónComisión Sectorial de Investigación CientíficaResearch Council United Kingdom Grand Challenges Research Funder (GCRF)Fondo para la Convergencia Estructural del Mercado Común del Sur (FOCEM)Pasteur Networ

Molecular Mechanisms of Vertical Transmission in Chagas Disease

Chagas disease is a parasitosis caused by the protozoan parasite Trypanosoma cruzi. It is estimated that almost 7 million infected people are worldwide. In nature, there are two forms of transmission: dependent on the triatomine vector, both by depositing faeces and introduction of the parasites and by ingestion (oral route) or independent of the vector through the transplacental route (vertical transmission); in addition, human activities such as blood transfusion, organ transplantation and laboratory accidents that constitute additional sources of infection. It causes high rates of morbidity and mortality in endemic and non-endemic areas, and there is currently a global spread of the disease due to migratory flows. Chagas disease is mainly transmitted by vertical transmission (TV) in non-endemic areas and in endemic areas where vector control programs have been successful. We have isolated and characterized natural strains of Trypanosoma cruzi transmitted vertically through three generations and proceeded to study the molecular mechanisms of VT, generating a murine model of vertical transmission. We have found that these strains are specialized for vertical transmission and behave completely differently from the reference strains. In this project, we propose to dissect the molecular mechanisms of VT through the study of the maternal-fetal interface, focusing on the one hand on the genetic characteristics of the isolates and, on the other, on the modulation of the immune response that allows control of the infection but not its transplacental transmission.Agencia Nacional de Investigación e InnovaciónInstitut Pasteur de Montevide

Pharmacological and Surgical Treatment for Trigeminal Neuralgia a Review of the Literature

Trigeminal neuralgia is the most common orofacial pain, with incidence levels that increase every year. Characterized by paroxysmal attacks of pain, brief, lancinating, stabbing, is considered one of the most painful conditions affecting patients. Drug therapy has been the gold standard for the management and control of this neuropathy, on the contrary, recent studies find wide drug resistance with history of neuralgia for many years, causing recurrence of the picture. That is why there is a need to establish a proper surgical planning, among which stands vascular decompression and percutaneous micro harmful methods such as retrogasserianas rhizotomies, chemical methods, such as injection of alcohol or glycerol in the oval hole, mental foramen, as an alternative to medical-pharmacological treatment, however, not all procedures reviewed in the literature indicate a continued success rate and further, the various adverse effects involving the implementation of each treatment should be considered

Increasing sales in a real estate company using 5S and standardization work: a case of study

The objective of this research was to analyze the results of a service company in the real estate sector in Peru, with the aim of improving the current sales process. The study began with an initial audit where it was possible to identify the causes of why the company wasn’t achieving more sales. These were the little order around work of the salesmen, a non-standardized sales process, and few control processes. These causes were identified in the diagnosis stage using engineering analysis tools such as the Ishikawa diagram, SIPOC, and Pareto diagram. After that, a model focused on the stages of the Deming cycle was used as a philosophical tool that gave us an order to apply the 5S tool and the Standardized Work tool; achieving the order of the offices, a standardized process, and finally, the implementation of simulation software named Arena let us measure the company results. After the implementation of the model, sales were projected to increase, the use of human resources was reduced by 8.54%, reprocesses in customer contact were eliminated and the percentage of sales efficiency grew by 65% compared to the leads received

Genome-wide chromatin interaction map for Trypanosoma cruzi

Trypanosomes are eukaryotic, unicellular parasites, such as Trypanosoma brucei, which causes sleeping sickness, and Trypanosoma cruzi, which causes Chagas disease. Genomes of these parasites comprise core regions and species-specific disruptive regions that encode multigene families of surface glycoproteins. Few transcriptional regulators have been identified in these parasites, and the role of spatial organization of the genome in gene expression is unclear. Here we mapped genome-wide chromatin interactions in T. cruzi using chromosome conformation capture (Hi-C), and we show that the core and disruptive regions form three-dimensional chromatin compartments named C and D. These chromatin compartments differ in levels of DNA methylation, nucleosome positioning and chromatin interactions, affecting genome expression dynamics. Our data reveal that the trypanosome genome is organized into chromatin-folding domains and transcription is affected by the local chromatin structure. We propose a model in which epigenetic mechanisms affect gene expression in trypanosomes.Agencia Nacional de Investigación e InnovaciónComisión Sectorial de Investigación Científica, Universidad de la RepúblicaResearch Council United Kingdom Grand Challenges Research Funder (GCRF)Fondo para la Convergencia Estructural del Mercado Común del Sur (FOCEM

Comparative microRNA profiling of Trypanosoma cruzi infected human cells

Introduction: Trypanosoma cruzi, the causative agent of Chagas disease, can infect almost any nucleated cell in the mammalian host. Although previous studies have described the transcriptomic changes that occur in host cells during parasite infection, the understanding of the role of post-transcriptional regulation in this process is limited. MicroRNAs, a class of short non-coding RNAs, are key players in regulating gene expression at the post-transcriptional level, and their involvement in the host-T. cruzi interplay is a growing area of research. However, to our knowledge, there are no comparative studies on the microRNA changes that occur in different cell types in response to T. cruzi infection. Methods and results: Here we investigated microRNA changes in epithelial cells, cardiomyocytes and macrophages infected with T. cruzi for 24 hours, using small RNA sequencing followed by careful bioinformatics analysis. We show that, although microRNAs are highly cell type-specific, a signature of three microRNAs -miR-146a, miR-708 and miR-1246, emerges as consistently responsive to T. cruzi infection across representative human cell types. T. cruzi lacks canonical microRNA-induced silencing mechanisms and we confirm that it does not produce any small RNA that mimics known host microRNAs. We found that macrophages show a broad response to parasite infection, while microRNA changes in epithelial and cardiomyocytes are modest. Complementary data indicated that cardiomyocyte response may be greater at early time points of infection. Conclusions: Our findings emphasize the significance of considering microRNA changes at the cellular level and complement previous studies conducted at higher organizational levels, such as heart samples. While miR-146a has been previously implicated in T. cruzi infection, similarly to its involvement in many other immunological responses, miR-1246 and miR-708 are demonstrated here for the first time. Given their expression in multiple cell types, we anticipate our work as a starting point for future investigations into their role in the post-transcriptional regulation of T. cruzi infected cells and their potential as biomarkers for Chagas disease

Early Trypanosoma cruzi Infection Triggers mTORC1-Mediated Respiration Increase and Mitochondrial Biogenesis in Human Primary Cardiomyocytes

Chagasic chronic cardiomyopathy is one of the most frequent and severe manifestations of Chagas disease, caused by the parasite Trypanosoma cruzi. The pathogenic and biochemical mechanisms responsible for cardiac lesions remain not completely understood, although it is clear that hypertrophy and subsequent heart dilatation is in part caused by the direct infection of cardiomyocytes. In this work, we evaluated the initial response of human cardiomyocytes to T. cruzi infection by transcriptomic profiling. Immediately after infection, cardiomyocytes dramatically change their gene expression patterns, up regulating most of the genes encoding for respiratory chain, oxidative phosphorylation and protein synthesis. We found that these changes correlate with an increase in basal and maximal respiration, as well as in spare respiratory capacity, which is accompanied by mitochondrial biogenesis pgc-1α independent. We also demonstrate that these changes are mediated by mTORC1 and reversed by rapamycin, resembling the molecular mechanisms described for the non-chagasic hypertrophic cardiomyopathy. The results of the present work identify that early during infection, the activation of mTORC1, mitochondrial biogenesis and improvement in oxidative phosphorylation are key biochemical changes that provide new insights into the host response to parasite infection and the pathogenesis of chronic chagasic cardiomyopathy. The finding that this phenotype can be reversed opens a new perspective in the treatment of Chagas disease, through the identification of host targets, and the use of combined parasite and host targeted therapies, in order to prevent chagasic cardiomyopathy

The tritryps comparative repeatome: insights on repetitive element evolution in trypanosomatid pathogens

The major human pathogens Trypanosoma cruzi, Trypanosoma brucei, and Leishmania major are collectively known as the Tritryps. The initial comparative analysis of their genomes has uncovered that Tritryps share a great number of genes, but repetitive DNA seems to be extremely variable between them. However, the in-depth characterization of repetitive DNA in these pathogens has been in part neglected, mainly due to the well-known technical challenges of studying repetitive sequences from de novo assemblies using short reads. Here, we compared the repetitive DNA repertories between the Tritryps genomes using genome-wide, low-coverage Illumina sequencing coupled to RepeatExplorer analysis. Our work demonstrates that this extensively implemented approach for studying higher eukaryote repeatomes is also useful for protozoan parasites like trypanosomatids, as we recovered previously observed differences in the presence and amount of repetitive DNA families. Additionally, our estimations of repetitive DNA abundance were comparable to those obtained from enhanced-quality assemblies using longer reads. Importantly, our methodology allowed us to describe a previously undescribed transposable element in Leishmania major (TATE element), highlighting its potential to accurately recover distinctive features from poorly characterized repeatomes. Together, our results support the application of this low-cost, low-coverage sequencing approach for the extensive characterization of repetitive DNA evolutionary dynamics in trypanosomatid and other protozoan genomes