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• We identified 169 circulating lineages across 16 South American countries. • SARS-CoV-2 lineage B is the most prevalent lineage in South America. • Emergent lineages of SARS-CoV2 are a plausible threat to the region. To evaluate the genomic diversity and geographic distribution of SARS-CoV-2 lineages in South America. SARS-CoV-2 lineages from a public dataset of 5583 South American genome assemblies were analyzed. Polymorphisms in the main open reading frames were identified and compared to those in the main lineages of epidemiological concern: B.1.1.7 (UK) and B.1.351 (South Africa). Across 16 South American countries, 169 lineages were identified; major lineage B had the greatest diversity and broadest geographic distribution. Seventeen predominant lineages were analyzed revealing 2 dominant lineages of concern: P.1 (Brazilian variant) and B.1.1.7 with 94 and 28 genomes, respectively, both with 33 polymorphisms (other lineages displayed ≤24 polymorphisms). A high number of polymorphisms were detected with a limited number of common variable positions, in common with the profile of the main lineages of epidemiological concern. The ever-increasing genetic diversity of SARS-CoV-2 continues to lead to novel lineage emergence. Various variants and lineages are now present across South America, dominated by major lineage B. The circulation of P.1 and B.1.1.7 and the high number of polymorphisms highlight the importance of genomic surveillance to determine introduction events, identify transmission chains, trace emergence, and implement prevention, vaccination and control strategies. [ABSTRACT FROM AUTHOR]

Urea is the main precursor of ethyl carbamate in fortified wines, which is in turn mostly produced by Saccharomyces cerevisiae due to the arginine catabolism during alcoholic fermentation. Due to its potential safety risks, efforts have been taken to reduce ethyl carbamate content by reducing the urea produced. However, most of them have been made through genetic manipulation, and their use in the food industry is therefore limited by legal constraints. In the present study, the adaptive laboratory evolution technique had been used to improve this trait in a diploid wine yeast already used at industrial level to obtain Sherry base wine. For this purpose, the genetic variability of the yeast population was increased by sexual reproduction and subsequently canavanine, a toxic arginine analogue, was applied as selective pressure to select yeast variants with lower urea production. Finally, an evolved variant that showed 62% lower urea content than the parental strain, also displaying an enhanced fermentative performance, was selected. The base Sherry wine obtained at industrial level not only showed a lower urea and ethyl carbamate content, but also an improvement in the aromatic profile, being fruitier and fresher than that obtained with the parental strain mainly due to an increase in ester content. • Urea and ethyl carbamate were reduced by adaptive evolution using l -canavanine. • Evolved strains upregulated the expression of permeases as well as urea amydolase. • Fermentation performance was enhanced by selected yeast variants. • Wine at industrial level showed better aromatic profile than with the parental one. [ABSTRACT FROM AUTHOR]

Gross alpha ( A α ) and gross beta activities ( A β ) were measured weekly in the airborne of the Universitat Politècnica de Valencia campus (in the east of Spain) during the period 2009–2015 (7 years). The geometric mean values of weekly A α and A β were 0.53·10 −4 Bq m −3 and 5.77·10 −4 Bq m −3 , respectively; with an average ratio A α / A β of 0.097. This study highlights the heterogeneity of gross alpha and gross beta activities depending on the different periods of the year. Data show seasonal variations with the highest activity in summer months and the lowest one in winter months. Several atmospheric factors were considered in order to explain this intra-annual variation (wind speed, temperature, relative humidity, precipitations, dust content and prevailing wind directions). Multiple Linear Regression Analysis were performed in order to obtain information on significant atmospheric factors that affect gross α and gross β variability, which could be useful in identifying meteorological or atmospheric changes that could cause deviations in gross α and gross β activity depending on the seasons considered. Models obtained explain more than 60% of variability for global data, and also for winter and spring-autumn months. However, more research is needed to explain gross α and gross β variability in summer months, because the atmospheric factors considered in the MLR explain less than 35% of variability. [ABSTRACT FROM AUTHOR]

Metacyclic trypomastigotes are essential for the understanding of the biology of Trypanosoma cruzi , the agent of Chagas disease. However, obtaining these biological stages in axenic medium is difficult. Techniques based on charge and density of the parasite during different stages have been implemented, without showing a high efficiency in the purification of metacyclic trypomastigotes. So far, there is no protocol implemented where sepharose-DEAE is used as a resin. Therefore, herein we tested its ability to purify metacyclic trypomastigotes in Liver Infusion Triptose (LIT) medium cultures. A simple, easy-to-execute and effective protocol based on ion exchange chromatography on Sepharose-DEAE resin for the purification of T . cruzi trypomastigotes is described. T . cruzi strains from the Discrete Typing Units (DTUs) I and II were used. The strains were harvested in LIT medium at a concentration of 1 × 10 7 epimastigotes/mL. We calculated the time of trypomastigotes increment (TTI). Based on the data obtained, Ion exchange chromatography was performed with DEAE-sepharose resin. To verify the purity and viability of the trypomastigotes, a culture was carried out in LIT medium with subsequent verification with giemsa staining. To evaluate if the technique affected the infectivity of trypomastigotes, in vitro assays were performed in Vero cells and in vivo in ICR-CD1 mice. The technique allowed the purification of metacyclic trypomastigotes of other stages of T . cruzi in a percentage of 100%, a greater recovery was observed in cultures of 12 days. There were differences regarding the recovery of metacyclic trypomastigotes for both DTUs, being DTU TcI the one that recovered a greater amount of these forms. The technique did not affect parasite infectivity in vitro or/and in vivo . [ABSTRACT FROM AUTHOR]

The recent increase in monkeypox (MPX) cases has attracted attention of public health authorities due to its quick spread and transmission across non-endemic regions. This outbreak, unlike previous ones, displays different epidemiological features and transmission dynamics, which appear to be largely influenced by the newly divergent MPX lineages (B.1). Yet, the genomic characteristics driving the high dispersal and diversification of these lineages remain largely unknown. Herein, we sought to explore and characterize the genomic features and phylogenetic diversity of the B.1 lineages through a comparative genomic analysis inclusive of 1900 high quality complete MPXV genomes. Our analyses indicate that the current MPXV-2022 outbreak encompasses thirteen derived lineages with ten unique non-synonymous mutations in several genes linked to immune evasion, virulence factors and host recognition. Such mutations may translate in the rapid evolution and diversification of current MPXV lineages. Moreover, our analyses uncovered signals of genomic modifications suggestive of immune-modulatory enzymatic activity, such as APOBEC3 editing, which, as previously suggested could have favored evolutionary trends leading to the rapid spread of MPXV into non-endemic countries. Genomic surveillance continues to play a major role in unveiling the genomic signatures signaling potential adaptation of this emerging MPXV lineage and how it will continue to impact public health in the near future. [ABSTRACT FROM AUTHOR]

Monkeypox is a zoonotic disease with clinical manifestations similar to smallpox in humans. Since May 13, 2022, an increasing number of suspected and confirmed cases have been reported, affecting non-endemic regions across the globe. More strikingly, reports from the current outbreak reveal unique aspects regarding transmission dynamics and an unprecedented, rapidly expanding and sustained community transmission. As demonstrated through the still-ongoing COVID-19 pandemic, genomic surveillance has been an essential resource for monitoring and tracking the evolution of pathogens of public health relevance. Herein, we performed a phylogenomic analysis of available Monkeypox virus (MPXV) genomes to determine their evolution and diversity. Our analysis revealed that all MPXV genomes grouped into three monophyletic clades: two previously characterized clades and a newly emerging clade harboring genomes from the ongoing 2022 multi-country outbreak with 286 genomes comprising the hMPXV-1A clade and the newly classified lineages: A.1 (n = 6), A.1.1 (n = 1), A.2 (n = 3) and B.1 (n = 262), where lineage B.1 includes all MPXV genomes from the 2022 outbreak. Finally, it was estimated that B.1 lineage of this clade emerged in Europe on 03/02/2022 [95%CI = 11/13/2021 to 05/10/2022]. The exceptional surge of cases and the broader geographical expansion suggest multifactorial factors as drivers of the current outbreak dynamics. Such factors may include the cessation of smallpox vaccination and its potential spread across particular networks. Integrating pertinent epidemiological information with genomic surveillance information will help generate real-time data to help implement adequate preventive and control measures by optimizing public health decisions to mitigate this outbreak. [ABSTRACT FROM AUTHOR]

Abstract: Objective: Growth hormone (GH) resistance leads to enhanced insulin sensitivity, decreased systolic blood pressure and increased lifespan. The aim of this study was to determine if there is a shift in the balance of the renin-angiotensin system (RAS) towards the ACE2/Ang-(1–7)/Mas receptor axis in the heart and the kidney of a model of GH resistance and retarded aging, the GH receptor knockout (GHR−/−) mouse. Design: RAS components were evaluated in the heart and the kidney of GHR−/− and control mice by immunohistochemistry and Western blotting (n=12 for both groups). Results: The immunostaining of Ang-(1–7) was increased in both the heart and the kidney of GHR−/− mice. These changes were concomitant with an increased immunostaining of the Mas receptor and ACE2 in both tissues. The immunostaining of AT1 receptor was reduced in heart and kidney of GHR−/− mice while that of AT2 receptor was increased in the heart and unaltered in the kidney. Ang II, ACE and angiotensinogen levels remained unaltered in the heart and the kidney of GH resistant mice. These results were confirmed by Western blotting and correlated with a significant increase in the abundance of the endothelial nitric oxide synthase in both tissues. Conclusions: The shift within the RAS towards an exacerbation of the ACE2/Ang-(1–7)/Mas receptor axis observed in GHR−/− mice could be related to a protective role in cardiac and renal function; and thus, possibly contribute to the decreased incidence of cardiovascular diseases displayed by this animal model of longevity. [Copyright &y& Elsevier]

Abstract: Angiotensin (Ang)-(1–7) stimulates proteins belonging to the insulin signaling pathway and ameliorates the Ang II negative effects at this level. However, up to date, receptors involved and mechanisms behind these observations remain unknown. Accordingly, in the present study, we explored the in vivo effects of antagonism of the Ang-(1–7) specific Mas receptor on insulin signal transduction in rat insulin-target tissues. We evaluated the acute modulation of insulin-stimulated phosphorylation of Akt, GSK-3β (Glycogen synthase kinase-3β) and AS160 (Akt substrate of 160kDa) by Ang-(1–7) and/or Ang II in the presence and absence of the selective Mas receptor antagonist A-779 in insulin-target tissues of normal rats. Also using A-779, we determined whether the Mas receptor mediates the improvement of insulin sensitivity exerted by chronic Ang-(1–7) treatment in fructose-fed rats (FFR), a model of insulin resistance, dyslipidemia and mild hypertension. The two major findings of the present work are as follows; 1) Ang-(1–7) attenuates acute Ang II-mediated inhibition of insulin signaling components in normal rats via a Mas receptor-dependent mechanism; and 2). The Mas receptor appears to be involved in beneficial effects of Ang-(1–7) on the phosphorylation of crucial insulin signaling mediators (Akt, GSK-3β and AS160), in liver, skeletal muscle and adipose tissue of FFR. These results shed light into the mechanism by which Ang-(1–7) exerts its positive physiological modulation of insulin actions in classical metabolic tissues and reinforces the central role of Akt in these effects. [Copyright &y& Elsevier]

Abstract: The in vivo effect of angiotensin (ANG)-(1-7) on the activation of insulin signaling transduction in rat extracardiac tissues is unknown. Thus, in the present study, we evaluated the ability of ANG-(1-7) to stimulate the phosphorylation of Akt, a main mediator of insulin action in rat extracardiac tissues (adipose tissue, liver and skeletal muscle). We proved that ANG-(1-7) induces the phosphorylation of Akt at both threonine 308 and serine 473 in all tissues analyzed. Selective antagonism of the Mas receptor with A779 blocked the ANG-(1-7)-induced Akt phosphorylation in extracardiac tissues. Reinforcing this evidence, we determined that ANG-(1-7) induces the in vivo activation of the downstream target of Akt, glycogen synthase kinase-3β in liver and skeletal muscle. Moreover, in every tissue analyzed, the presence of the Mas receptor was detected by immunohistochemical analysis. Based on the current results, we postulate that ANG-(1-7) could be a positive physiological contributor to the actions of insulin in extracardiac tissues. Therefore, our findings extend the possibilities for new approaches in the study of ANG-(1-7)/Mas receptor axis and show the therapeutic potential of ANG-(1-7) in the treatment of metabolic disorders such as insulin resistance as well as other disorders associated with diminished Akt activity. [Copyright &y& Elsevier]

Abstract: Context: Growth hormone (GH) is an anabolic hormone that regulates growth and metabolism. Ames dwarf mice are natural mutants for Prop1, with impaired development of anterior pituitary and undetectable levels of circulating GH, prolactin and TSH. They constitute an endocrine model of life-long GH-deficiency. The main signaling cascades activated by GH binding to its receptor are the JAK2/STATs, PI-3K/Akt and the MAPK Erk1/2 pathways. Objectives: We have previously reported that GH-induced STAT5 activation was higher in Ames dwarf mice liver compared to non-dwarf controls. The aim of this study was to evaluate the principal components of the main GH-signaling pathways under GH-deficiency in liver and skeletal muscle, another GH-target tissue. Methods: Ames dwarf mice and their non-dwarf siblings were assessed. Animals were injected i.p. with GH or saline 15min before tissue removal. Protein content and phosphorylation of signaling mediators were determined by immunoblotting of tissue solubilizates. Results: GH was able to induce STAT5 and STAT3 tyrosine phosphorylation in both liver and muscle, but the response was higher for Ames dwarf mice than for non-dwarf controls. When Erk1/2 activation was assessed in liver, only dwarf mice showed GH-induced phosphorylation, while in muscle no response to the hormone was found in either genotype. GH-induced Akt phosphorylation at Ser473 in liver was only detected in dwarf mice. In skeletal muscle, both normal and dwarf mice responded to a GH stimulus, although dwarf mice presented higher GH activation levels. The phosphorylation of GSK-3, a substrate of Akt, increased upon hormone stimulation only in dwarf mice in both tissues. In contrast, no differences in the phosphorylation of mTOR, another substrate of Akt, were observed after GH stimulus, either in normal or dwarf mice in liver, while we were unable to determine mTOR in muscle. Protein content of GH-receptor and of the signaling mediators studied did not vary between normal and dwarf animals in the assessed tissues. Conclusion: These results show that several components of the main GH-signaling pathways exhibit enhanced sensitivity to the hormone in liver and muscle of Ames dwarf mice. [Copyright &y& Elsevier]

The objective of this study was to determine the effects of high-pressure treatments and mild temperatures on endogenous microflora and Escherichia coli CECT 515 artificially inoculated into orange and apple juices and vegetable soup. In general, the viability of aerobic bacteria was significantly reduced as pressure and temperature increased. Although the greatest reduction in the concentration of aerobic mesophilic vegetative cells was reached at 350 MPa and 60°C, the same reduction occurred in fruit juices at 350 MPa and 20°C. Yeasts and molds were below the level of detection (1 log CFU/ml) for the fruit juices and did not exceed 2 log CFU/ml for vegetable soup. Foods inoculated with E. coli were subjected to several treatments as indicated by the mathematical model applied in response surface methodology to obtain the maximum information with the minimum number of experiments. The number of tests for a range of pressures (150 to 350 MPa) and temperatures (20 to 60°C) was limited to 11. The models were considered adequate because of satisfactory R² values. The optimum process parameters (pressure and temperature) for a 6-log reduction of E. coli were obtained at 248.25 MPa and 59.91°C in orange juice, 203.50 MPa and 57.18°C in apple juice, and 269.8 MPa and 59.9°C in vegetable soup. Sublethal injury of E. coli occurred as pressure and temperature increased. Nearly all of the E. coli cells were injured at 350 MPa and 20°C in fruit juices and after all treatments in vegetable soup. [ABSTRACT FROM AUTHOR]

The objective of this study was to compare the effects of combined high hydrostatic pressure and temperature treatments with different chemical sanitation treatments (water, sodium hypochlorite, and hydrogen peroxide) on the microbiological properties of mung bean sprouts. In a first study, the raw product was subjected to several combined high-pressure and temperature treatments for calculating a mathematical model by a response surface methodology. The number of pressure- temperature (150 to 400 MPa; 20 to 40°C) combinations was limited to 10. In addition, a model system consisting of mung bean sprout juice was inoculated with Listeria monocytogenes (CECT 4032). Microbial inactivation with this model system was also investigated by a response surface methodology. The highest aerobic mesophilic bacteria and L. monocytogenes inactivation was achieved at maximum pressure and temperature (5.5 and 1.8 log cycles, respectively). In a second study, the effect of five different processing lines on the microbial load reduction of minimally processed mung bean sprouts during refrigerated storage was studied. All treatments reduced the initial population of aerobic mesophilic bacteria and fecal coliforms, with the physical treatment of 400 MPa and 40°C being the most effective, showing initial reductions of 5.8 and 7.8 log CFU/g, respectively. Recovery of bacteria from sprouts treated under these conditions was not observed during storage. However, the sprouts that received washing treatments with water, sodium hypochlorite, and hydrogen peroxide exhibited increases in aerobic mesophilic and fecal coliform counts after 3 days of storage at 4°C. [ABSTRACT FROM AUTHOR]

Abstract: Growth hormone (GH) is an important modulator of insulin sensitivity. Multiple mechanisms appear to be involved in this modulatory effect. GH does not interact directly with the insulin receptor (IR), but conditions of GH excess are associated in general with hyperinsulinemia that induces a reduction of IR levels and impairment of its kinase activity. Several post-receptor events are shared between GH and insulin. This signaling crosstalk could be involved in the diabetogenic effects of GH. The utilization of animal models of GH excess, deficiency or resistance provided evidence that the signaling pathway leading to stimulation of the phosphatidylinositol 3-kinase (PI3K)/Akt cascade is an important site of regulation, and pointed to the liver as the major site of GH-induced insulin resistance. In skeletal muscle, GH-induced insulin resistance might involve an increase in the amount of the p85 subunit of PI3K that plays a negative role in insulin signalling. GH also reduces insulin sensitivity by enhancing events that negatively modulate insulin signaling such as stimulation of serine phosphorylation of IRS-1, which prevents its recruitment to the IR and induction of the suppressor of cytokine signalling (SOCS)-1 and SOCS-3 which modulate the signalling potential of the IRS proteins. In addition, GH has been shown to decrease the expression of the insulin-sensitizing adipo-cytokines adiponectin and visfatin. Finally, genetic manipulation of mice indicated that whereas GH plays a major role in reducing insulin sensitivity, circulating IGF-I also participates in the control of insulin sensitivity and plays an important role in the hormonal balance between GH and insulin. [Copyright &y& Elsevier]

Venezuela and Colombia both adopted measures of containment early in response to the COVID-19 pandemic. However, Venezuela's ongoing humanitarian crisis has decimated its health care system, and forced millions of Venezuelans to flee through its porous border with Colombia. The extensive shared border, and illegal cross-border transit through improvised trails between the two countries are major challenges for public health authorities. We report the first SARS-CoV-2 genomes from Venezuela, and present a snapshot of the SARS-CoV-2 epidemiologic landscape in the Colombian-Venezuelan border region. We sequenced and assembled viral genomes from total RNA extracted from nasopharyngeal (NP) clinical specimens using a custom reference-based analysis pipeline. Three assemblies obtained were subjected to typing using the Phylogenetic Assignment of Named Global Outbreak LINeages 'Pangolin' tool. A total of 376 publicly available SARS-CoV-2 genomes from South America were obtained from the GISAID database to perform comparative genomic analyses. Additionally, the Wuhan-1 strain was used as reference. We found that two of the SARS-CoV-2 genomes from Venezuela belonged to the B1 lineage, and the third to the B.1.13 lineage. We observed a point mutation in the Spike protein gene (D614G substitution), previously reported to be associated with increased infectivity, in all three Venezuelan genomes. Additionally, three mutations (R203K/G204R substitution) were present in the nucleocapsid (N) gene of one Venezuelan genome. Genomic sequencing demonstrates similarity between SARS-CoV-2 lineages from Venezuela and viruses collected from patients in bordering areas in Colombia and from Brazil, consistent with cross-border transit despite administrative measures including lockdowns. The presence of mutations associated with increased infectivity in the 3 Venezuelan genomes we report and Colombian SARS-CoV-2 genomes from neighboring borders areas may pose additional challenges for control of SARS-CoV-2 spread in the complex epidemiological landscape in Latin American countries. Public health authorities should carefully follow the progress of the pandemic and its impact on displaced populations within the region. • SARS-CoV-2 genomes obtained from Venezuela reveal the presence of B1 and B.1.13 lineages, suggesting two separate introductions. • All Venezuelan genomes carried a D614G substitution in the spike (S) protein, which has been linked to increased infectivity. • Three substitutions in the nucleocapsid (N) gene were additionally identified in one of the examined genomes. [ABSTRACT FROM AUTHOR]

This paper presents a three-stage protocol for gross alpha and gross beta evaluation in water samples in emergencies. The novelty of this approach is the great level of detail for its application in this type of sample, following the criteria proposed in well-established safety guidelines. This protocol makes use of a rapid method adapted from different proposals found in the scientific literature. The method is based on a simple preparation of the sample and a rapid measurement by liquid scintillation counting on Quantulus 1220, which permits the evaluation of waters with different salt content (from 5 g l−1 of continental and drinking water, to 35 g l−1 of seawater) and pH (from 1 to 8) in emergency situations. The protocol and the method allow to prioritize the most active samples and to assess contamination in less than 2 h. Both were tested and validated with spiked water samples with different ratios of alpha and beta emitters (1:1, 1:10 and 10:1) and with intercomparison water samples. Relative bias are below 10%, except in the samples with activities close to the limits of detection and relative standard deviation are below 10% in most of the samples, which give a clear idea of the robustness of the method. • Rapid method for gross α and gross β evaluation in water samples. • Evaluation of waters with different salt content (5–35 g l−1) and pH (1-8). • Emergency response protocol to prioritize active samples and evaluate contamination. • Rapid and robust method with low relative bias and relative standard deviations. [ABSTRACT FROM AUTHOR]

• Visceral leishmaniasis can occur due to co-infection with several Leishmania spp. • Co-infection in visceral leishmaniasis changes our understanding of the disease. • Several trypanosomatids can be identified per sample using this protocol. • Amplicon-based next-generation sequencing is a cost-effective method in molecular epidemiological studies. Visceral leishmaniasis (VL) is a mammalian protozoal disease propagated in the Americas by female phlebotomine sandflies, mainly caused by Leishmania infantum. However, in recent years, cases of VL caused by different Leishmania species, such as L. amazonensis and L. colombiensis, have been reported in the continent. This study used an amplicon-based next-generation sequencing approach to identify VL aetiologic species using high-depth sequencing targeting a region on the Heat Shock Protein 70 gene. In this first approach, six samples from five patients diagnosed with VL were selected and analysed to identify DNA of Leishmania spp. All samples harboured DNA of L. infantum ; five samples were found to be co-infected with other Leishmania spp. or with Trypanosoma cruzi , and just one sample was mono-infected with L. infantum. This study demonstrates the usefulness of this methodology to identify trypanosomatid co-infections in clinical samples, which presents an interesting study panorama considering their biological, clinical and epidemiological implications. [Display omitted] [ABSTRACT FROM AUTHOR]

• SARS-CoV-2 lineages were identified among Venezuelan migrants in Colombia. • The spike L18F mutation was identified in genomes from Venezuelan migrants. • Colombian and Venezuelan isolates from the northern border were genetically similar. • Within Colombian departments, Venezuelan and Colombian isolates were distinct. • This is the first report to describe the presence of B.1.1.255 lineage in Colombia. To evaluate the genomic epidemiology of SARS-CoV-2 from Venezuelan migrants living in Colombia. This study sequenced SARS-CoV-2 from 30 clinical specimens collected from Venezuelan migrants. Genomes were compared with the Wuhan reference genome to identify polymorphisms, reconstruct phylogenetic relationships and perform comparative genomic analyses. Geographic, sociodemographic and clinical data were also studied across genotypes. This study demonstrated the presence of six distinct SARS-CoV-2 lineages circulating among Venezuelan migrants, as well as a close relationship between SARS-CoV-2 genomic sequences obtained from individuals living in the Venezuelan-Colombian border regions of La Guajira (Colombia) and Zulia (Venezuela). Three clusters (C-1, C-2 and C-3) were well supported by phylogenomic inference, supporting the hypothesis of three potential transmission routes across the Colombian-Venezuelan border. These genomes included point mutations previously associated with increased infectivity. A mutation (L18F) in the N-terminal domain of the spike protein that has been associated with compromised binding of neutralizing antibodies was found in 2 of 30 (6.6%) genomes. A statistically significant association was identified with symptomatology for cluster C2. The close phylogenetic relationships between SARS-CoV-2 genomes from Venezuelan migrants and from people living at the Venezuela-Colombian border support the importance of human movements for the spread of COVID-19 and for emerging virus variants. [ABSTRACT FROM AUTHOR]

Clostridioides difficile may have a negative impact on gut microbiota composition in terms of diversity and abundance, thereby triggering functional changes supported by the differential presence of genes involved in significant metabolic pathways, such as short-chain fatty acids (SCFA). This work has evaluated shotgun metagenomics data regarding 48 samples from four groups classified according to diarrhea acquisition site (community- and healthcare facility-onset) and positive or negative Clostridioides difficile infection (CDI) result. The metagenomic-assembled genomes (MAGs) obtained from each sample were taxonomically assigned for preliminary comparative analysis concerning differences in composition among groups. The predicted genes involved in metabolism, transport, and signaling remained constant in microbiota members; characteristic patterns were observed in MAGs and genes involved in SCFA butyrate and acetate metabolic pathways for each study group. A decrease in genera and species, as well as relative MAG abundance with the presence of the acetate metabolism-related gene, was evident in the HCFO/- group. Increased antibiotic resistance markers (ARM) were observed in MAGs along with the genes involved in acetate metabolism. The results highlight the need to explore the role of acetate in greater depth as a potential protector of the imbalances produced by CDI, as occurs in other inflammatory intestinal diseases. [ABSTRACT FROM AUTHOR]

Abstract: Identifying the minimal functional regions of the proteins which the malaria parasite uses when invading its host cells constitutes the first and most important approach in an effective design for a chemically synthesised, multi-antigen, multi-stage, subunit-based vaccine. This work has been aimed at identifying the PfRh1 protein binding regions (residues 1–2580) belonging to the reticulocyte binding-like (RBL or P. falciparum Rh [PfRh]) family implicated in the parasite''s alternative target cell invasion routes. Eighteen peptide regions (called high activity binding peptides – HABPs) binding to red blood cells (RBC) were identified in peptides mapped in a highly robust, specific and sensitive receptor–ligand assay. These HABPs were saturable in the experimental conditions assayed here and most had an alpha helix structure. Polymorphism studies revealed that only six of the eighteen HABPs identified had changes at amino acid level amongst the seven P. falciparum strains evaluated. Most HABPs’ specific binding became altered when RBC were treated with neuraminidase, chymotrypsin and trypsin, suggesting differing sensitivity for RBC membrane receptors. After ascertaining that the Rh1 gene was transcribed and expressed in late-stage schizonts of the FCB-2 strain, invasion inhibition assays were carried out. When most of these HABPs were assayed in P. falciparum in vitro culture they were able to inhibit high percentages of FVO strain invasion compared to low inhibition percentages observed with the FCB-2 strain. This data shows small Rh1 regions’ participation during invasion and suggests that these units should be included in further immunological and structural studies. [Copyright &y& Elsevier]

Abstract: Identifying the ligands or regions derived from them which parasites use to invade their target cells has proved to be an excellent strategy for identifying targets for vaccine development. Members of the reticulocyte-binding homologue family (PfRH), including RH5, have been implicated in invasion as adhesins binding to specific receptors on erythrocyte surface. The regions mediating PfRH5-RBC specific interactions have been identified here by fine mapping the whole PfRH5 protein sequence. These regions, called high activity binding peptides (HABPs), bind to a receptor which is sensitive to trypsin treatment and inhibit merozoite invasion of RBCs by up to 80%, as has been found for HABP 36727. Our results show that a single amino acid change in the HABP 36727 sequence modifies a peptide''s 3D structure, thereby resulting in a loss of specific binding to human RBCs and its inhibition ability, while binding to Aotus RBC remains unmodified. Such invasion differences and binding ability produced by replacing a single amino acid in an essential molecule, such as PfRH5, highlight the inherent difficulties associated with developing a fully effective vaccine against malaria. [Copyright &y& Elsevier]

Abstract: Two recently described molecules have been associated with sporozoite traversal ability and hepatocyte entry: sporozoite invasion-associated proteins (SIAP)-1 and -2. The HeLa and HepG2 cell binding ability of synthetic peptides spanning the whole SIAP-1 and -2 sequences has been studied in the search for identifying these proteins’ functionally active specific regions. Twelve HepG-2 and seventeen HeLa cell high-activity binding peptides (HABPs) have been identified in SIAP-1, 8 of them having high specific binding affinity for both cell lines. Four HepG2 HABPs and two HeLa HABPs have been identified in SIAP-2, one of them interacting with both HeLa and HepG2 cells. SIAP-1 and SIAP-2 HABPs bound specifically and saturably to heparin sulfate and chondroitin sulfate-type membrane receptors on host cells. Circular dichroism assays have shown high α-helix content in SIAP-1 and SIAP-2 HABP secondary structure. Immunofluorescence analysis has revealed that specific peptides against SIAP proteins are highly immunogenic in mice and that anti-SIAP-1 and -2 antibodies recognize the native protein in Plasmodium falciparum sporozoites. Polymorphism studies have shown that a most SIAP-1 and -2 HABPs are conserved among P. falciparum strains. Our results have suggested that SIAP-1 and -2 participate in host–pathogen interactions during cell-traversal and hepatocyte invasion and highlighted the relevance of the ongoing identification and study of potentially new molecules when designing a fully protective antimalarial vaccine. [Copyright &y& Elsevier]

Abstract: Ames dwarf mice are a model of retarded aging and extended longevity and display enhanced insulin sensitivity. Caloric restriction (CR) and the dwarf mutation have additive effects on lifespan. To begin to understand the mechanisms behind this effect, an analysis of the in vivo status of the insulin signaling system was performed in skeletal muscle from Ames dwarf (df/df) and normal mice fed ad libitum or subjected to long-term (over 1 year) CR. The response to CR was different in both groups of animals. In normal animals, CR induced a significant reduction in both circulating insulin and glucose levels, together with an increase in the in vivo insulin-stimulated phosphorylation of the IR, a trend towards an increase in the in vivo insulin-stimulated phosphorylation levels of IR substrate-1, and an increase in the abundance of GLUT4 in muscle. In contrast, CR did not modify none of these parameters in df/df mice. Interestingly, CR induced a reduction in the p85 subunit of phosphatidylinositol 3-kinase abundance in skeletal muscle in both groups of animals. These results suggest that in skeletal muscle, long-term CR induces different effects on the first steps of the insulin signaling system in normal mice than in df/df mice. [Copyright &y& Elsevier]

• The Trypanosoma cruzi parasitic load is not life-stage dependent. • Epimastigotes can be used to quantify T. cruzi parasitic loads. • Molecular diagnostics of T. cruzi must be improved. Chagas disease is a complex zoonosis caused by Trypanosoma cruzi. The diagnosis of this infection is complex and molecular tools are suggested to detect the parasite in blood samples. A long-standing question arises in Chagas disease molecular diagnostics and is related to the feasibility of using epimastigotes in standard curves to quantify parasitic loads. Herein, we conducted experiments running standard curves with all the known life stages of T. cruzi. Our results indicate that regardless of the life stage employed, there are no statistically significant differences when calculating parasitic loads in blood samples. Our results have practical implications from a laboratory perspective in terms of the usability of epimastigotes to build standard curves for T. cruzi pan-stage assessment. Future studies are needed to further improve T. cruzi molecular diagnostic methods and enhance their impact in clinical practice. [ABSTRACT FROM AUTHOR]

Leishmania amazonensis is one of the causative agents of the different forms of cutaneous leishmaniasis present in Latin America. This species has been isolated from humans and animals (canine/feline) in some endemic regions of Colombia. Therefore, L. amazonensis is of great relevance at the clinical and epidemiological levels in medicine and veterinary science. Until now, very few genomes from this species are available. Here, we report the complete genome sequence of a laboratory-adapted L. amazonensis strain isolated from a human patient with clinical manifestation of cutaneous leishmaniasis in Colombia. The genome sequence not only allowed inter and intra-species comparative analyses to be performed with the sequenced genomes of L. amazonensis strains from different geographical regions, but also increased our knowledge about the genomic behavior of this L. amazonensis Colombian strain. This isolate was also characterized in terms of single nucleotide polymorphisms, chromosome and gene copy number variations (CNVs). The results revealed moderate aneuploidy, CNVs in genes involved in the virulence, growth, and survival of the parasite, and in the distributions of the multicopy genes on some chromosomes, as well as a high level of heterozygosity. The data confirmed previous reports that identified unique variations in L. amazonensis , suggesting aneuploidy may not have a high fitness cost and may allow the rapid generation of diversity in Leishmania parasites growing normally. [ABSTRACT FROM AUTHOR]

Clostridium perfringens is a gram-positive, anaerobic sporulating bacillus which can infect several hosts, thereby being considered the causative agent of many gut illnesses. Some studies have suggested that C. perfringens 's virulence factors may negatively affect gut microbiota homeostasis by decreasing beneficial bacteria; however, studies have failed to evaluate the simultaneous presence of other pathogenic bacteria, such as C. difficile (another sporulating bacillus known to play a role in gut microbiota imbalance). Conscious of the lack of compelling data, this work has ascertained how such microorganisms' coexistence can be associated with a variation in gut microbiota composition, compared to that of C. perfringens colonisation. PCR was thus used for identifying C. perfringens and C. difficile in 98 samples. Amplicon-based sequencing of 16S- and 18S-rRNA genes' V4 hypervariable region from such samples was used for determining the microbiota's taxonomical composition and diversity. Small differences were observed in bacterial communities' taxonomic composition and diversity; such imbalance was mainly associated with groups having hospital-acquired diarrhoea. The alterations reported herein may have been influenced by C. difficile and diarrhoea acquisition site, despite C. perfringens ' ability to cause alterations in microbiota due to its virulence factors. Our findings highlight the need for a holistic view of gut microbiota. • C. perfringens is linked to increased Bacteroidota and decreased Saccharomycetes. • C. perfringens and C. difficile coexist associated with hospital-acquired diarrhea. • Relationships among microbiota members highlight the need to analyze interactions. [ABSTRACT FROM AUTHOR]

Diet is known to strongly modulate the composition of the gut microbiota, thereby affecting health conditions and disease. Natural BARF-type and commercial diets have been used for feeding pets (e.g. dogs and cats) promoting changes in the canine microbiota in terms of abundance, richness, and diversity that may favor certain metabolic processes and resistance to certain infectious agents. Therefore, the present study sought to identify microbiota changes in dogs fed with a BARF-type diet versus dogs fed with a commercial diet by sequencing the V4 region of the 16S rRNA gene. The microbiota of dogs fed with the BARF-diet (n = 20) and commercial-diet (n = 26) was studied using fecal samples. A metabarcoding strategy was employed by sequencing the V4 hypervariable region of the 16S rRNA gene using the Illumina HiSeq platform. DADA2 was used to assess the quality profile of the reads and to determine the core sample inference algorithm of the reads to infer amplicon sequence variants (ASVs). The taxonomic assignment was performed using sequences from the Silva v138 formatted reference database. The microbial diversity analysis was performed using the R package Phyloseq, which was used to calculate diversity and abundance indices and construct the respective graphs. Linear discriminant analysis (LDA) effect size analysis (LEfSe) was used to identify the differentially abundant taxa in the BARF group versus the commercial-diet group. The diet causes changes in fecal microbiota composition and diversity, with richness and diversity being higher in BARF-fed dogs. Beta diversity analyses confirmed that diet is directly related to microbiota composition regardless of breed or sex. Differentially enriched taxa were identified in each of the diets as Fusobacterium , Bacteroides, and Clostridium perfringens in BARF-fed dogs and Prevotella , Turicibacter , Faecalibacterium, and Peptacetobacter (Clostridium) hiranonis , mostly relevant in carbohydrate metabolism, in commercial-fed dogs. This study is the first one carried out in dogs from Colombia that seeks to identify changes in the intestinal microbiota concerning natural BARF type diet and commercial diet using a metabarcoding approach. Important differences were identified in terms of richness, diversity, and differentially enriched bacteria in each of the diets. The microbiota of dogs fed the BARF diet was characterized by higher richness and diversity compared to the commercial diet. However, it was identified that BARF-fed dogs can potentially acquire more opportunistic infections by pathogens of importance such as C. perfringens. Most of the taxa enriched in commercial diet-fed dogs are linked to carbohydrate metabolism, which may be directly related to diet composition. • Diet strongly modulates gut microbiota in dogs. • BARF diet increases richness and diversity. • Commercial diet enriched bacteria linked to carbohydrate metabolism. • Fusobacterium, Bacteroides, C. perfringens enriched in BARF. • Prevotella, Turicibacter, Faecalibacterium, Peptacetobacter enriched in commercial. [ABSTRACT FROM AUTHOR]

Transmission of cutaneous leishmaniasis in Venezuela reveals diverse and changing epidemiological landscapes, as well as a spectrum of clinical phenotypes presumed to be linked to a variety of Leishmania species. Central-western Venezuela constitutes one of the highest endemic epicenters in the country, and updated molecular epidemiological information is still lacking. Therefore, in this study we aimed to characterize the landscape of circulating Leishmania species across central-western Venezuela through the last two decades, performed comparisons of haplotype and nucleotide diversity, and built a geospatial map of parasite species distribution. A total of 120 clinical samples were collected from patients across the cutaneous disease spectrum, retrieving parasitic DNA, and further characterizing by PCR and sequencing of the HSP70 gene fragment. This data was later collated with further genetic, geospatial and epidemiological analyses. A peculiar pattern of species occurrence including Leishmania (Leishmania) amazonensis (77.63% N=59), Leishmania (Leishmania) infantum (14.47% N=11), Leishmania (Viannia) panamensis (5.26% N=4) and Leishmania (Viannia) braziliensis (2.63% N=2) was revealed , also highlighting a very low genetic diversity amongst all analyzed sequences. Geographical distribution showed that most cases are widely distributed across the greater urban-sub urban area of the Irribaren municipality. L.(L.) amazonensis appears to be widely dispersed throughout Lara state. Statistical analyses failed to reveal significance for any comparisons, leading to conclude a lack of association between the infective Leishmania species and clinical phenotypes. To the best of our knowledge, this is an unprecedented study which addresses comprehensively the geographical distribution of Leishmania species in central-western Venezuela throughout the last two decades, and the first to incriminate L. (L.) infantum as an etiologic agent of cutaneous leishmaniasis in this region. Our findings support that Leishmania endemism in central-western Venezuela is caused mainly by L.(L.) amazonensis. Future studies are needed to unveil additional details on the ecological intricacies and transmission aspects of leishmaniasis (i.e. sampling phlebotomines and mammals) and to adopt adequate public health prevention and control strategies and mitigate disease impact in this endemic region. [ABSTRACT FROM AUTHOR]

Abstract: The effect of minimal processing on the health-related attributes of orange fruit was investigated. Oranges were prepared as whole fruits, hand-peeled fruits and manually separated segments, packed under air atmosphere and stored at 4°C for 12days. The stability of main bioactive compounds (carotenoids, flavanones and vitamin C) and antioxidant activity was evaluated. The total carotenoid content showed a significant increase for the whole samples during refrigerated storage, whereas no significant changes were observed for segments or peeled samples. A similar trend was found for vitamin A. With regard to vitamin C, at the end of refrigerated storage, some losses were observed although no significant differences were found among the different processed samples. The flavanone content showed a significant increase throughout refrigerated storage as response to cold stress. In general, the antioxidant activity remained stable in relation to the initial values. Hence, the health-related characteristics of minimally processed oranges were retained during refrigerated storage. [ABSTRACT FROM AUTHOR]

Abstract: The aim of the present study was to determine if insulin can modulate the pressor response to angiotensin II at brain level in normotensive rats. Anaesthetized male rats were intracerebroventricularly infused with insulin (12mU/h, n =15) or Ringer’s solution as vehicle (n =15) for 2h. Immediately, changes in mean arterial pressure (MAP) in response to an intracerebroventricular subpressor dose of angiotensin II (5pmol, n =10) or vehicle (n =5) were measured for 10min. Then, hypothalami were removed and Akt and ERK1/2 phosphorylation levels were determined. In other subset of animals, PD98059 (MAPK inhibitor) or vehicle were intracerebroventricularly administered previously to insulin perfusion for 2h and changes in MAP in response to intracerebroventricular angiotensin II (5pmol) injection were evaluated for 10min (n =6 for each group). Angiotensin II did not modify MAP in vehicle pre-treated rats, but increased MAP in insulin pre-treated animals. Insulin significantly increased Akt phosphorylation, but no changes were observed after angiotensin II injection in vehicle-pretreated animals. Angiotensin II or insulin infusion increased in more than two fold phospho-ERK 1/2 hypothalamic levels. Animals that received insulin infusion followed by Ang II injection presented 4.5 higher values than those which received vehicle, and nearly twice than those who received Ang II without insulin pre-treatment. PD98059 administration abolished the blood pressure response exerted by angiotensin II in insulin pre-treated rats. In conclusion, centrally administered insulin potentiates the pressor effects to angiotensin II, suggesting a novel mechanism, possibly involving MAPK activation, by which insulin influences blood pressure control at central level. [Copyright &y& Elsevier]

The crucial role played by Ag163B6/cruzipain, the major cystein proteinase of Trypanosoma cruzi, in the process of parasite internalization into mammalian cells and IgG hydrolysis, signals this antigen as a potential target for raising a protective immune response against Chagas’ disease. On the other hand, synthetic oligodeoxynucleotides containing CpG-motifs (CpG-ODN) are capable of driving immunity toward a Th1 bias. Considering the importance of Th1 mechanisms in resistance against this intracellular parasite, we analyzed the ability of Ag163B6/cruzipain plus CpG-ODN to induce immunoprotection against a lethal challenge with trypomastigotes. Mice were immunized with Ag163B6+CpG-ODN showing high specific antibody titers, mostly IgG2a. Spleen cells from these mice strongly proliferated and presented significant increase of IL-2 and IFN-γ concentrations in their supernatant upon antigen stimulation. Trypomastigote challenge rendered elevated parasitemia and mortality in all control groups, meanwhile Ag163B6+CpG-ODN mice displayed the lowest level of blood parasites and 100% survival to acute infection. Besides, we demonstrated that other parasite antigens introduced into mice when challenged, and consequently never seen before by the immune system, also elicited a Th1 immune response. Taken together, these results plus others provide the basis for the design of a multicomponent anti-T. cruzi vaccine which may ultimately be used not only to protect humans at risk of infection, but also may alleviate or prevent the pathogenic responses characteristic of chronic Chagas’ disease by reducing or perhaps eliminating tissue parasites from infected patients. [Copyright &y& Elsevier]

The renin-angiotensin system modulates insulin action. Pharmacological stimulation of angiotensin type 2 receptor (AT2R) was shown to have beneficial metabolic effects in various animal models of insulin resistance and type 2 diabetes and also to increase insulin sensitivity in wild type mice. In this study we further explored the role of the AT2R on insulin action and glucose homeostasis by investigating the glycemic profile and in vivo insulin signaling status in insulin-target tissues from both male and female AT2R knockout (KO) mice. When compared to the respective wild-type (WT) group, glycemia and insulinemia was unaltered in AT2RKO mice regardless of sex. However, female AT2RKO mice displayed decreased insulin sensitivity compared to their WT littermates. This was accompanied by a compensatory increase in adiponectinemia and with a specific attenuation of the activity of main insulin signaling components (insulin receptor, Akt and ERK1/2) in adipose tissue with no apparent alterations in insulin signaling in either liver or skeletal muscle. These parameters remained unaltered in male AT2RKO mice as compared to male WT mice. Present data show that the AT2R has a physiological role in the conservation of insulin action in female but not in male mice. Our results suggest a sexual dimorphism in the control of insulin action and glucose homeostasis by the AT2R and reinforce the notion that pharmacological modulation of the balance between the AT1R and AT2R receptor could be important for treatment of metabolic syndrome and type 2 diabetes. • AT2RKO mice present a sexual dimorphism in terms of insulin sensitivity. • AT2R deletion impairs insulin tolerance in female mice. • AT2RKO does not affect insulin sensitivity in male mice. • In female mice, AT2R deficiency leads to a selective attenuation of insulin signaling in adipose tissue. • AT2R in adipose tissue appears to have a major role in the control of insulin action. [ABSTRACT FROM AUTHOR]