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Van Wyk-Grumbach syndrome is a rare form of severe hypothyroidism. We present a 10.9-year-old girl who consulted for genital bleeding, Tanner stage 2, and clinical manifestations of hypothyroidism. Severe hypothyroidism was confirmed, secondary to chronic lymphocytic thyroiditis. A pelvic ultrasound showed bilateral evidence of cystic ovarian masses. The hormonal profile confirmed peripheral precocious puberty secondary to hypothyroidism. A pituitary MRI showed significant pituitary elongation due to thyrotropic hyperplasia. After initiating treatment with thyroid hormone, there was a notable clinical improvement and resolution of the adnexal masses and pituitary hyperplasia. The pediatrician must be able to identify signs and symptoms associated with thyroid dysfunction and prompt referral to a pediatric endocrinologist to avoid the appearance of severe conditions such as the one presented., (Sociedad Argentina de Pediatría.)

The 'sacred leaf' or "Hoja Santa" (Piper auritum Kunth) has a great value for Mexican culture and has gained popularity worldwide for its excellent properties from culinary to remedies. To contribute to its heritage, in this project we proposed the green synthesis of silver oxide nanoparticles (Ag 2 O NPs) using an extract of "Hoja Santa" (Piper auritum) as a reducing and stabilizing agent. The synthesized Ag 2 O NPs were characterized by UV-Visible spectroscopy (plasmon located at 405 nm), X-ray diffraction (XRD) (particle size diameter of 10 nm), scanning electron microscopy (SEM) (particle size diameter of 13.62 ± 4.61 nm), and Fourier-transform infrared spectroscopy (FTIR) (functional groups from "Hoja Santa" attached to nanoparticles). Antioxidant capacity was evaluated using DPPH, ABTS and FRAP methods. Furthermore, the antimicrobial activity of NPs against a panel of clinically relevant bacterial strains, including both Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Salmonella Enteritidis and Escherichia coli O157:H7), was over 90% at concentrations of 200 µg/mL. Additionally, we assessed the antibiofilm activity of the NPs against Pseudomonas aeruginosa (reaching 98% of biofilm destruction at 800 µg/mL), as biofilm formation plays a crucial role in bacterial resistance and chronic infections. Moreover, we investigated the impact of Ag 2 O NPs on immune cell viability, respiratory burst, and phagocytic activity to understand their effects on the immune system., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Patagonia (Argentina and Chile) harbors the highest Saccharomyces eubayanus genomic diversity and its widest predominance in natural environments. In this work, S. eubayanus was isolated for the first time from a fermentative environment. This species was found dominating both a traditional apple chicha fermentation as well as feral apple trees in the Andean region of Aluminé (Argentina). S. eubayanus was the only Saccharomyces species found in the isolation substrates, although it coexisted with other non-Saccharomyces species. The absence of strong fermentative competitors of the Saccharomyces genus (like Saccharomyces uvarum or Saccharomyces cerevisiae) in the feral apples could promote the development and implantation of S. eubayanus in a spontaneous apple must fermentation. Phylogeographic analyses revealed a high intraspecific diversity in S. eubayanus, enabling the characterization of strains belonging to the genomic subpopulations PA1, PA2, and PB1 according to the sequences obtained for the intFR gene region. This result evidence that the studied sampling area represents a natural habitat for the species. Being a novel finding, studying the causes that allowed this species to prosper in a fermentative environment becomes essential. Hence, the physiological profile of the new isolates, including their ability to grow at different temperature, nitrogen, and ethanol concentrations was evaluated in comparison with a set of S. eubayanus strains previously isolated from natural environment and representing different genomic subpopulations. Greater physiological diversity was evidenced when strains isolated from both natural and fermentative environments were analyzed overall. Furthermore, no direct relationship between genomic population and physiological behavior was observed; on the opposite, strains appeared to exhibit similar behavior, primarily grouped by isolation origin., (© 2023 John Wiley & Sons Ltd.)

Introduction. During hospitalization, patients may develop significant clinical deterioration and require unplanned admission to the pediatric intensive care unit (PICU). This may result in increased morbidity and mortality. These events are often preceded by a deterioration phase that may go unnoticed. Objective. To determine the frequency, analyze the causes, and describe the clinical characteristics and outcomes of unplanned transfers of hospitalized pediatric patients from the general pediatric ward (GPW) to the PICU, and analyze the differences between urgent and emergent transfers. Population and methods. Prospective, descriptive study; all unplanned transfers from the GPW to the PICU occurring between January 1st , 2014 and December 31st, 2019 were analyzed. Results. There were 212 unplanned transfers (21 transfers per 1000 admissions). An associated comorbidity was present in 76% of transferred patients -being cancer the most frequent one (36%)- and they had been hospitalized for more than 24 hours in the GPW. The most frequent causes of transfer were respiratory distress (43%), sepsis (20%), and neurological/neurosurgical complications (20%). The overall mortality rate was 8.96% (19 patients). Conclusions. The analysis of unplanned transfers is a critical component in the assessment of the quality of care and patient safety of an area, and should be an indicator integrated into the control panel. The interpretation of unplanned transfers as a preventable event is a key paradigm shift., (Sociedad Argentina de Pediatría.)

The wild relatives of rice hold unexplored genetic diversity that can be employed to feed an estimated population of 10 billion by 2050. The Oryza Map Alignment Project (OMAP) initiated in 2003 has provided comprehensive genomic resources for comparative, evolutionary, and functional characterization of the wild relatives of rice, facilitating the cloning of >600 rice genes, including those for grain width (GW5) and submergence tolerance (SUB1A). Following in the footsteps of the original project, the goal of 'IOMAP: the Americas' is to investigate the present and historic genetic diversity of wild Oryza species endemic to the Americas through the sequencing of herbaria and in situ specimens. The generation of a large diversity panel describing past and current genetic status and potential erosion of genetic variation in the populations will provide useful knowledge for the conservation of the biodiversity in these species. The wild relatives of rice in the Americas present a wide range of resistance traits useful for crop improvement and neodomestication approaches. In the race against time for a sustainable food future, the neodomestication of the first cereal species recently accomplished in O. alta opens the door to the potential neodomestication of the other wild Oryza species in Americas., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

Different species can find convergent solutions to adapt their genome to the same evolutionary constraints, although functional convergence promoted by chromosomal rearrangements in different species has not previously been found. In this work, we discovered that two domesticated yeast species, Saccharomyces cerevisiae, and Saccharomyces uvarum, acquired chromosomal rearrangements to convergently adapt to the presence of sulfite in fermentation environments. We found two new heterologous chromosomal translocations in fermentative strains of S. uvarum at the SSU1 locus, involved in sulfite resistance, an antimicrobial additive widely used in food production. These are convergent events that share similarities with other SSU1 locus chromosomal translocations previously described in domesticated S. cerevisiae strains. In S. uvarum, the newly described VIIXVI and XIXVI chromosomal translocations generate an overexpression of the SSU1 gene and confer increased sulfite resistance. This study highlights the relevance of chromosomal rearrangements to promote the adaptation of yeast to anthropic environments., Competing Interests: The authors have declared that no competing interests exist.

Willows are widely planted in areas under risk of flooding. The physiological responses of willows to flooding have been characterized, but little is known about their responses during the post-flooding period. After the end of the stress episode, plants may modify some traits to compensate for the biomass loss during flooding. The aim of this work was to analyze the post-flooding physiological responses of willow under two different depths of stagnant floodwater. Cuttings of Salix matsudana NZ692 clone were planted in pots in a greenhouse. The experiment started when the plants were 2 months old with the following treatments: Control plants (watered to field capacity); plants partially flooded 10 cm above soil level (F10) and plants partially flooded 40 cm above soil level (F40). The flooding episode lasted 35 days and was followed by a recovery period of 28 days (post-flooding period). After the flooding period, height, diameter and total biomass were higher in F10, while F40 plants showed an increase in plant adventitious root production and leaf nitrogen content. During the post-flooding period, the photosynthetic rate, nitrogen, chlorophyll and soluble sugar contents were significantly higher in leaves of F40 than in Control and F10 treatments. Stomatal conductance and specific leaf area were higher in the previously flooded plants compared to Control treatment. Plants from F10 treatments showed a higher growth in height, root-to-shoot ratio, and carbon isotope discrimination than F40, while the opposite occurred for growth in diameter, vessel size and leaf area. We conclude that depth of floodwater not only causes different responses during flooding, but that its effects are also present in the post-flooding recovery period, affecting the growth and physiology of willows once the stress episode has ended. Even when flooding impacted growth negatively in F40, in the post-flooding period these plants compensated by increasing the photosynthetic rate, plant leaf area and xylem vessel size. Willows endurance to flooding is the result of both responses during flooding, and plastic responses during post-flooding., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Mozo, Rodríguez, Monteoliva and Luquez.)

Solea senegalensis aquaculture production has experienced a great increase in the last decade and, consequently, the genome knowledge of the species is gaining attention. In this sense, obtaining a high-density genome mapping of the species could offer clues to the aquaculture improvement in those aspects not resolved so far. In the present article, a review and new processed data have allowed to obtain a high-density BAC-based cytogenetic map of S. senegalensis beside the analysis of the sequences of such BAC clones to achieve integrative data. A total of 93 BAC clones were used to localize the chromosome complement of the species and 588 genes were annotated, thus almost reaching the 2.5% of the S. senegalensis genome sequences. As a result, important data about its genome organization and evolution were obtained, such as the lesser gene density of the large metacentric pair compared with the other metacentric chromosomes, which supports the theory of a sex proto-chromosome pair. In addition, chromosomes with a high number of linked genes that are conserved, even in distant species, were detected. This kind of result widens the knowledge of this species' chromosome dynamics and evolution.

Knowing the factors responsible for sex determination in a species has significant theoretical and practical implications; the dmrt1 gene (Doublesex and Mab-3 (DM)-related Transcription factor 1) plays this role in diverse animal species. Solea senegalensis is a commercially important flat fish in which females grow 30% faster than males. It has 2n = 42 chromosomes and an XX / XY chromosome system for sex determination, without heteromorph chromosomes but with sex proto-chromosome. In the present study, we are providing the genomic structure and nucleotide sequence of dmrt1 gene obtained from cDNA from male and female adult gonads. A cDNA of 2027 containing an open-reading frame (ORF) of 1206 bp and encoding a 402 aa protein it is described for dmrt1 gene of S. senegalensis. Multiple mRNA isoforms indicating a high variable system of alternative splicing in the expression of dmrt1 of the sole in gonads were studied. None isoforms could be related to sex of individuals. The genomic structure of the dmrt1 of S. senegalensis showed a gene of 31400 bp composed of 7 exons and 6 introns. It contains an unexpected duplication of more than 10399 bp, involving part of the exon I, exons II and III and a SINE element found in the sequence that it is proposed as responsible for the duplication. A mature miRNA of 21 bp in length was localized at 336 bp from exon V. Protein-protein interacting networks of the dmrt1 gene showed matches with dmrt1 protein from Cynoglossus semilaevis and a protein interaction network with 11 nodes (dmrt1 plus 10 other proteins). The phylogenetic relationship of the dmrt1 gene in S. senegalensis is consistent with the evolutionary position of its species. The molecular characterization of this gene will enhance its functional analysis and the understanding of sex differentiation in Solea senegalensis and other flatfish., Competing Interests: The authors have declared that no competing interests exist.

The flatfish, Solea senegalensis has considerable scientific interest and commercial value. The metamorphosis in this species occurs between 12 and 19 days after hatching and it takes about 1 week to complete. Eleven Bacterial Artificial Chromosomes (BAC) clones containing the various candidate genes involved in the process of metamorphosis: thyroxine 5 deiodinase 3 (dio3); forkhead box protein E4 (foxe4); melatonin receptor type 1C (mel1c); calsequestrin 1b (casq1b); thyrotropin subunit beta (tshβ); thyrotropin-releasing hormone receptor 1, 2, and 3 (trhr1, trhr2, trhr3); thyroid hormone receptor α a and b (thrαa, thrαb); and thyroid hormone receptor beta (thrβ) were analyzed by multiple Fluorescence in situ Hybridization (mFISH) and Next Generation Sequencing (NGS) techniques. The mFISH technique localized the 11 BAC clones on 12 different chromosome pairs because three of them, specifically the trhr1a, trhr2 and thrβ BAC clones, showed double signals. This signal duplication indicates a duplication of the genomic region inserted within the BAC clone, which provides evidence for the Teleost-Specific Whole Genome Duplication (TS-WGD). Micro-synteny and phylogenetic analysis showed that Cynoglossus semilaevis is the nearest species to S. senegalensis and that Danio rerio is the most distant one. The tshβ BAC clone was highly conserved as the genes belonging to this BAC were located on a single chromosome in all the species studied. These genes participate in proliferation, migration and cell-death, which are key processes during metamorphosis. Overall, micro-synteny analysis showed that most candidate genes are found in conserved genomic surroundings., Competing Interests: Declaration of competing interest The authors declare that they do not have any conflict of interest., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Purpose: To examine effects of a 10-week after-school physical activity (PA) program on academic performance of 6- to 12-year-old African American children with behavior problems., Methods: Participants were randomized to PA (n = 19) or sedentary attention control (n = 16) programs. Academic records, curriculum-based measures, and classroom observations were obtained at baseline, postintervention, and/or follow-up. Mixed models tested group × time interactions on academic records and curriculum-based measures. One-way analysis of variance or Kruskal-Wallis tested for differences in postintervention classroom observations., Results: Intent-to-treat analyses demonstrated a moderate effect within groups from baseline to postintervention on disciplinary referrals (PA: d = -0.47; attention control: d = -0.36) and a null moderate effect on academic assessments (PA: d = 0.11 to 0.36; attention control: d = 0.05 to 0.40). No significant group × time interactions emerged on direct academic assessments (all Ps ≥ .05, d = -0.23 to 0.26) or academic records (all Ps ≥ .05, d = -0.28 to 0.16). Classroom observations revealed that intervention participants were off-task due to moving at twice the rate of comparative classmates (F = 15.74, P < .001) and were off-task due to talking 33% more often (F = 1.39, P = .257)., Conclusion: Academic outcome improvements were small within and between groups and did not sustain at follow-up. Academic benefits of after-school PA programs for children with attention-deficit hyperactivity disorder and/or disruptive behavior disorders were smaller than neurobiological, behavioral, and cognitive outcomes as previously reported.

The wild, cold-adapted parent of hybrid lager-brewing yeasts, Saccharomyces eubayanus, has a complex and understudied natural history. The exploration of this diversity can be used both to develop new brewing applications and to enlighten our understanding of the dynamics of yeast evolution in the wild. Here, we integrate whole genome sequence and phenotypic data of 200 S. eubayanus strains, the largest collection known to date. S. eubayanus has a multilayered population structure, consisting of two major populations that are further structured into six subpopulations. Four of these subpopulations are found exclusively in the Patagonian region of South America; one is found predominantly in Patagonia and sparsely in Oceania and North America; and one is specific to the Holarctic ecozone. Plant host associations differed between subpopulations and between S. eubayanus and its sister species, Saccharomyces uvarum. S. eubayanus is most abundant and genetically diverse in northern Patagonia, where some locations harbor more genetic diversity than is found outside of South America, suggesting that northern Patagonia east of the Andes was a glacial refugium for this species. All but one subpopulation shows isolation-by-distance, and gene flow between subpopulations is low. However, there are strong signals of ancient and recent outcrossing, including two admixed lineages, one that is sympatric with and one that is mostly isolated from its parental populations. Using our extensive biogeographical data, we build a robust model that predicts all known and a handful of additional regions of the globe that are climatically suitable for S. eubayanus, including Europe where host accessibility and competitive exclusion by other Saccharomyces species may explain its continued elusiveness. We conclude that this industrially relevant species has rich natural diversity with many factors contributing to its complex distribution and natural history., Competing Interests: We have read the journal's policy and the authors of this manuscript have the following competing interests: commercial use of Saccharomyces eubayanus strains requires a license from WARF or CONICET. Strains are available for academic research under a material transfer agreement.

Climate change will increase the risk of flooding in several areas of the world where Populus deltoides Marshall (eastern cottonwood) is planted, so it would be desirable for this species to select for flooding tolerance. The aims of this work were to explore the variability in growth, leaf traits and flooding tolerance in an F1 full-sib intraspecific progeny of P. deltoides, to analyze the correlations of leaf and growth traits with flooding tolerance and to assess their suitability for use in breeding programs. Two-month-old parental clones and their progeny of 30 full-sib F1 genotypes were grown in pots and subjected to two treatments: (i) plants watered to field capacity (control) and (ii) plants flooded up to 10 cm above soil level for 35 days. Growth (height, diameter and biomass partition) and leaf traits (leaf size and number, specific leaf area, leaf senescence, abscission, stomatal conductance, carbon isotope discrimination, stomatal index) were measured. Flooding tolerance for each genotype was estimated as the ratio of the biomass of stressed plants to the biomass of control plants. Results showed segregation in terms of flooding tolerance in the F1 progeny. A significant genotype effect was found for leaf size and number, carbon isotopic discrimination and stomatal conductance, but it did not correlate with flooding tolerance. Height, diameter and root-to-shoot ratio had a positive phenotypic correlation with flooding tolerance, and there was a positive genetic correlation of height and diameter with biomass on both treatments. The narrow sense heritability values for the traits analyzed ranged from 0 to 0.56. We conclude that growth traits are more adequate than leaf traits for selection to increase flooding tolerance. A vigorous initial growth would increase flooding tolerance in young poplar plants., (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permission@oup.com.)

Repetitive sequences play an essential role in the structural and functional evolution of the genome, particularly in the sexual chromosomes. The Senegalese sole (Solea senegalensis) is a valuable flatfish in aquaculture albeit few studies have addressed the mapping and characterization of repetitive DNA families. Here we analyzed the Simple Sequence Repeats (SSRs) and Transposable elements (TEs) content from fifty-seven BAC clones (spanning 7.9 Mb) of this species, located in chromosomes by multiple fluorescence in situ hybridization (m-BAC-FISH) technique. The SSR analysis revealed an average density of 675.1 loci per Mb and a high abundance (59.69%) of dinucleotide coverage was observed, being 'AC' the most abundant. An SSR-FISH analysis using eleven probes was also carried out and seven of the 11 probes yielded positive signals. 'AC' probes were present as large clusters in almost all chromosomes, supporting the bioinformatic analysis. Regarding TEs, DNA transposons (Class II) were the most abundant. In Class I, LINE elements were the most abundant and the hAT family was the most represented in Class II. Rex/Babar subfamily, observed in two BAC clones mapping to chromosome pair 1, showed the longest match. This chromosome pair has been recently reported as a putative sexual proto-chromosome in this species, highlighting the possible role of the Rex element in the evolution of this chromosome. In the Rex1 phylogenetic tree, the Senegalese sole Rex1 retrotransposon could be associated with one of the four major ancient lineages in fish genomes, in which it is included O. latipes.

Global aquaculture production continues to increase rapidly. One of the most important species of marine fish currently cultivated in Southern Europe is Solea senegalensis , reaching more than 300 Tn in 2017. In the present work, 14 Bacterial Artificial Chromosome (BAC) clones containing candidate genes involved in the immune system ( b2m , il10 , tlr3 , tap1 , tnf α, tlr8 , trim25 , lysg , irf5 , hmgb2 , calr , trim16 , and mx ), were examined and compared with other species using multicolor Fluorescence in situ Hybridization (mFISH), massive sequencing and bioinformatic analysis to determine the genomic surroundings and syntenic chromosomal conservation of the genomic region contained in each BAC clone. The mFISH showed that the groups of genes hmgb2-trim25-irf5-b2m ; tlr3-lysg ; tnfα-tap1 , and il10-mx-trim16 were co-localized on the same chromosomes. Synteny results suggested that the studied BACs are placed in a smaller number of chromosomes in S. senegalensis that in other species. Phylogenetic analyses suggested that the evolutionary rate of immune system genes studied is similar among the taxa studied, given that the clustering obtained was in accordance with the accepted phylogenetic relationships among these species. This study contributes to a better understanding of the structure and function of the immune system of the Senegalese sole, which is essential for the development of new technologies and products to improve fish health and productivity.

Background: The re-sequencing of C. angulata has revealed many polymorphisms in candidate genes related to adaptation to abiotic stress that are not present in C. gigas; these genes, therefore, are probably related to the ability of this oyster to retain high concentrations of toxic heavy metals. There is, in addition, an unresolved controversy as to whether or not C. angulata and C. gigas are the same species or subspecies. Both oysters have 20 metacentric chromosomes of similar size that are morphologically indistinguishable. From a genomic perspective, as a result of the great variation and selection for heterozygotes in C. gigas, the assembly of its draft genome was difficult: it is fragmented in more than seven thousand scaffolds., Results: In this work sixty BAC sequences of C. gigas downloaded from NCBI were assembled in BAC-contigs and assigned to BACs that were used as probes for mFISH in C. angulata and C. gigas. In addition, probes of H3, H4 histone, 18S and 5S rDNA genes were also used. Hence we obtained markers identifying 8 out the 10 chromosomes constituting the karyotype. Chromosomes 1 and 9 can be distinguished morphologically. The bioinformatic analysis carried out with the BAC-contigs annotated 88 genes. As a result, genes associated with abiotic adaptation, such as metallothioneins, have been positioned in the genome. The gene ontology analysis has also shown many molecular functions related to metal ion binding, a phenomenon associated with detoxification processes that are characteristic in oysters. Hence the provisional integrated map obtained in this study is a useful complementary tool for the study of oyster genomes., Conclusions: In this study 8 out of 10 chromosome pairs of Crassostrea angulata/gigas were identified using BAC clones as probes. As a result all chromosomes can now be distinguished. Moreover, FISH showed that H3 and H4 co-localized in two pairs of chromosomes different that those previously escribed. 88 genes were annotated in the BAC-contigs most of them related with Molecular Functions of protein binding, related to the resistance of the species to abiotic stress. An integrated genetic map anchored to the genome has been obtained in which the BAC-contigs structure were not concordant with the gene structure of the C. gigas scaffolds displayed in the Genomicus database.

Background: Solea senegalensis (Kaup, 1858) is a commercially important flatfish species, belonging to the Pleuronectiformes order. The taxonomy of this group has long been controversial, and the karyotype of the order presents a high degree of variability in diploid number, derived from chromosomal rearrangements such as Robertsonian fusions. Previously it has been proposed that the large metacentric chromosome of S. senegalensis arises from this kind of chromosome rearrangement and that this is a proto-sex chromosome., Results: In this work, the Robertsonian origin of the large metacentric chromosome of S. senegalensis has been tested by the Zoo-FISH technique applied to two species of the Soleidae family (Dicologlossa cuneata and Dagetichthys lusitanica), and by comparative genome analysis with Cynoglossus semilaevis. From the karyotypic analysis we were able to determine a chromosome complement comprising 2n = 50 (FN = 54) in D. cuneata and 2n = 42 (FN = 50) in D. lusitanica. The large metacentric painting probe gave consistent signals in four acrocentric chromosomes of the two Soleidae species; and the genome analysis proved a common origin with four chromosome pairs of C. semilaevis. As a result of the genomic analysis, up to 61 genes were annotated within the thirteen Bacterial Artificial Chromosome clones analysed., Conclusions: These results confirm that the large metacentric chromosome of S. senegalensis originated from a Robertsonian fusion and provide new data about the chromosome evolution of S. senegalensis in particular, and of Pleuronectiformes in general.

Saccharomyces eubayanus is the close relative of the Lager-brewing yeast and was firstly found in North Patagonia associated with Nothofagus trees. In recent years additional strains were found in North America, Asia and New Zealand, and genomic analyses showed the existence of two main populations of this yeast, both of them present in Patagonia. Here, we performed the most comprehensive study of S. eubayanus in Patagonia natural environments (400 samples) and confirmed that this region has the highest isolation success rate for this species described worldwide (more than 10-fold). The genetic characterization of 200 isolates (COX2, DCR1, intFR) revealed five geographically structured subpopulations. We hypothesized that marine ingressions and glaciations, which shaped the Patagonian landscape, contributed on population differentiation. The first large screening of fermentation performance of 60 wild S. eubayanus strains indicated which subpopulations would be more suitable for beer production., (© 2018 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Terroir concept includes specific soil, topography, climate, landscape characteristics and biodiversity features. In reference to the last aspect, recent studies investigating the microbial biogeography (lately called 'microbial terroir') have revealed that different wine-growing regions maintain different microbial communities. The aim of the present work was to identify potential autochthonous fermentative yeasts isolated from native plants in North Patagonia, Schinus johnstonii, Ephedra ochreata and Lycium chilense, that could be associated to the specific vitivinicultural terroir of this region. Different Pichia kudriavzevii isolates were recovered from these plants and physiologically and genetically compared to regional wine isolates and foreign reference strains of the same species. All isolates were subjected to molecular characterization including mtDNA-RFLP, RAPD-PCR and sequence analysis. Both wine and native P. kudriavzevii isolates from Patagonia showed similar features, different from those showed by foreign strains, suggesting that this species could be part of a specific regional terroir from North Patagonia., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Introduction: Information on insulin reference values and insulin sensitivity indices in the field of pediatrics is scarce., Objective: To describe insulin range and insulin sensitivity surrogate indices during childhood., Materials and Methods: Fasting insulin level range and surrogate indices, such as the homeostasis model assessment of insulin resistance (HOMA-IR), among healthy children and adolescents by age, body mass index, pubertal stage (PS), insulin-like growth factor-1 (IGF-1), total cholesterol, and triglycerides., Results: Two hundred and twenty-six healthy children and adolescents (1-18 years old) were included. Insulin increased with age, body mass index, pubertal stage, IGF-1 and triglyceride levels (r2= 0.38, p 〈 0.0001). Prepubertal children 〉 7.5 years old had higher insulin levels [median (P3 and P97) pIU/mL: 5.0 (1.7-9.6)] than prepubertal children 〈 7.5 years old [2.9 pIU/ mL (1.3-10.9), p 〈 0.01]. During puberty (from PS II to PS V), insulin was higher in girls than in boys [7.4 (1.8-16.9) versus 5.8 (1.8-12.9), p 〈 0.01]. The HOMA-IR index increased in the group of prepubertal children 〉 7.5 years old: 1.1 (0.32.0) versus children 〈 7.5 years old: 0.6 (0.3-1.4, p 〈 0.01). The insulin level and HOMA-IR results were higher in pubertal children compared to the prepubertal group (p 〈 0.001)., Conclusions: Known physiological changes were observed inboth insulin levels and the HOMA-IR index among children and adolescents. A fasting blood insulin level of 10 pIU/mL in prepubertal children and of 17 pIU/mL and 13 pIU/mL in pubertal girls and boys, respectively, may be considered as an acceptable cut-off value in healthy children. A HOMA-IR value 〉 2.0 and 〉 2.6 in prepubertal and pubertal children, respectively, may be considered a warning sign for pediatricians to further investigate insulin resistance., (Sociedad Argentina de Pediatría.)

Objective: To assess whether sleep quality (SQ) at baseline is associated with improvement in pain and disability at 3 months., Materials and Methods: Four hundred twenty-two subacute and chronic patients with neck pain (NP) were recruited in 32 physiotherapy, primary care, and specialized centers. NP, referred pain, disability, catastrophizing, depression, and SQ were assessed through validated questionnaires, upon recruitment and 3 months later. Correlations between baseline scores were calculated through the Spearman coefficient. Improvements in NP, disability, and SQ were defined as a reduction of ≥30% of baseline score. Six estimative logistic regression models were developed to assess the association between baseline SQ and improvement of NP, baseline SQ and improvement of disability, baseline NP and improvement of SQ, baseline disability and improvement of SQ, the evolutions of NP and SQ, and the evolutions of disability and SQ., Results: Most patients were subacute and mildly impaired. Regression models showed that better SQ at baseline was associated with improvement of NP (odds ratio=0.91 [95% confidence interval, 0.83-0.99]), but not disability (1.04 [0.95-1.13]); the improvement of SQ was associated with more severe NP at baseline (1.26 [1.07-1.49)], but not with baseline disability (0.99 [0.97-1.02]); and that improvement in SQ was associated with improvements in NP (3.48 [1.68-7.20]), and disability (5.02 [2.39-10.11])., Discussion: NP is less likely to improve in patients with poorer SQ, irrespective of age, sex, catastrophizing, depression, or treatments prescribed for NP. Future studies should confirm these results with more severely impaired patients.

Here we describe the whole genome re-sequencing of the Portuguese oyster Crassostrea angulata, an edible cupped oyster of major commercial importance with an important role as biosensor of coastal water pollution. We sequenced the genome of the C. angulata to 29.3-fold coverage using ABI SOLID system. Comparisons of the sequences with the reference assembly of the Pacific oyster (Crassostrea gigas), yielded 129 million SNPs, 151,620 from which were located in 20,908 genes from the C. gigas database. The analysis of Gene Ontology (GO) terms associated with gene regions containing SNPs, revealed that significant GO terms showing differences between the two oyster species, were related to activities of response to stress caused both by drying and by metal contamination. In the Biological Process domain, the GO terms ion transport, phosphorylation and proteolysis processes, among others, showed many polymorphic genes in C. angulata. These processes are related to combating genotoxic and hypo-osmotic stress in the oyster. It is noteworthy that more than 200 polymorphic genes were associated with DNA repair processes. These results reveal that most of the gene polymorphisms observed in C. angulata are associated with processes related to genome adaptation to abiotic stress in estuarine regions and support that genetic polymorphisms may be the base to the observed ability of C. angulata to retain the phenomenally high concentrations of toxic heavy metals. Our results also provide the framework for future investigations to establish the molecular basis of phenotypic variation of adaptive traits and should contribute to the management of the species' genetic resources.

The effect of β-1,3/1,6-glucan, derived from yeast, on growth, antioxidant, and digestive enzyme performance of Pacific red snapper Lutjanus peru before and after exposure to lipopolysaccharides (LPS) was investigated. The β-1,3/1,6-glucan was added to the basal diet at two concentrations (0.1 and 0.2 %). The treatment lasted 6 weeks, with sampling at regular intervals (0, 2, 4, and 6 weeks). At the end of this period, the remaining fish from either control or β-glucan-fed fish were injected intraperitoneally with LPS (3 mg kg(-1)) or with sterile physiological saline solution (SS) and then sampled at 0, 24, and 72 h. The results showed a significant increase (P < 0.05) in growth performance after 6 weeks of feeding with β-glucan. Superoxide dismutase (SOD) activity in liver was significantly higher in diets containing 0.1 % β-glucan in weeks 4 and 6, compared to the control group. β-Glucan supplementation at 0.1 and 0.2 % significantly increased aminopeptidase, trypsin, and chymotrypsin activity. At 72 h after injection of LPS, we observed a significant increase in catalase activity in liver from fish fed diets supplemented with 0.1 and 0.2 % β-glucan; SOD activity increased in fish fed with 0.1 % β-glucan in relation to those injected with SS. Feed supplemented with β-1,3/1,6-glucan increased growth, antioxidant activity, and digestive enzyme activity in Pacific red snapper.

Bacterial LPS (endotoxin) has been implicated in the pathogenesis of acute liver disease through its induction of the proinflammatory cytokine TNF-α. TNF-α is a key determinant of the outcome in a well-established mouse model of acute liver failure during septic shock. One possible mechanism for regulating TNF-α expression is through the control of protein elongation during translation, which would allow rapid cell adaptation to physiological changes. However, the regulation of translational elongation is poorly understood. We found that expression of p38γ/δ MAPK proteins is required for the elongation of nascent TNF-α protein in macrophages. The MKK3/6-p38γ/δ pathway mediated an inhibitory phosphorylation of eukaryotic elongation factor 2 (eEF2) kinase, which in turn promoted eEF2 activation (dephosphorylation) and subsequent TNF-α elongation. These results identify a new signaling pathway that regulates TNF-α production in LPS-induced liver damage and suggest potential cell-specific therapeutic targets for liver diseases in which TNF-α production is involved.

The aim of this study was to identify the presence of Entamoeba histolytica and E. dispar by nested PCR in children attending the "Dr. Luis Razetti" Hospital, Barcelona, Anzoátegui State. Of the 1,141 fecal samples coproparasitologically evaluated by conventional microscopy, 150 were diagnosed positive for E. histolytica in 0-10 year-old-children, of both sexes. The signs, symptoms and a full coproparasitological report were obtained from all of these and nested PCR was performed to identify E. histolytica and E. dispar. The conventional microscopy results showed a diagnostic frequency of E. histolytica in 13.2% of the cases, of which 79.3% were positive only for this pathogen. However, nested PCR showed that of these, only 28% (42/150) were actually infected by Entamoeba spp., revealing a high over-diagnosis of E. histolytica. We also identified 9.3% E. histolytica, 4% E. dispar and 4.7% mixed infections. Diarrhea was the most common symptom, followed by abdominal pain and fever. Bloody stools were statistically associated with E. histolytica, but were also found for E. dispar infections. This study demonstrates that molecular techniques complementary to conventional methods enable the correct identification of Entamoeba spp., thus contributing to an improved epidemiological assessment of these parasites and implementation of the appropriate treatment.

Objective: To evaluate the temporal distribution (1991-2009) and associated variation of KSHV subtypes in Cuba., Method: Phylogenetic characterization based on the KSHV K1 gene was performed using 90 KSHV positive samples., Results: Molecular characterization confirmed the prevalence of a wide range of KSHV subtypes (A: n=48 [A5=12]; C: n=15; B: n=22; and E: n=5). In the current study, we observed a significant increase in HHV-8 subtype B after 2004 (p=0.0063). This Subtype B in Cuba was associated with: heterosexual behaviour (OR: 3.63, CI: 1,2-10,98; p=0.03), with the antecedent of acquiring HIV/KSHV in Africa (p=0.0003), with nodular stage of KS lesions (OR 4.2, CI: 1.1 to 15.7; p=0.04)., Conclusion: Our study is the first to report KSHV Subtype B expansion in Cuba, that might be reflective of a change in human behavioural pattern., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Objective: To assess the agreement between repeated blood pressure (BP) measurements taken by community pharmacists during visits to the community pharmacy on four separate occasions, and to assess the agreement between the community pharmacy BP (CPBP) measurement method and ambulatory BP monitoring (ABPM) in order to determine the CPBP measurements from each visit needed to evaluate the hypertensive state of treated patients., Methods: The study was a cross-sectional study of treated hypertensive patients aged older than 18 years from eight community pharmacies. BP was measured during four visits to the community pharmacy (three measurements per visit) and for 24 h using ABPM (ABPM started on the third visit to the pharmacy). The Lin correlation-concordance coefficient, the Bland-Altman method, and the κ coefficient were used to assess the agreement between CPBP measurements from each visit. The κ coefficient was used to assess the agreement between the CPBP measurement method and ABPM to establish the patient's hypertensive state. The average CPBP was calculated using four methods: (i) three measurements/four visits, (ii) two measurements/four visits, (iii) three measurements/three visits, and (iv) two measurements/three visits., Results: One hundred and seventy-six treated hypertensive patients were included. Overall, the agreement between the second and third CPBP measurement at each visit was higher than the agreement between other paired measurements (first vs. third and first vs. second), for both systolic BP and diastolic BP. The agreement between the CPBP measurement method and ABPM was not statistically different when analyzed using all variations of the average CPBP calculation., Conclusion: The agreement between repeated BP measurements taken by community pharmacists at each visit was greater between the second and the third measurements. In contrast, the agreement between the CPBP measurement method and ABPM did not improve when the first CPBP measurement that was taken at each visit was excluded. As a result, the three CPBP measurements taken at each visit to the pharmacy could be used to evaluate the hypertensive state of treated patients.

In Cuba, previous reports have shown an increase of epidemic KS, reaching a total of 120 cases by the end of 2007, despite the use of HAART. To evaluate and compare the role of human herpes virus 8 (HHV-8) viral loads in different compartments of AIDS-related Kaposi's sarcoma (AIDS-KS) patients real-time polymerase chain reaction (RT-PCR) was used to determine the genome copy number of HHV-8 in plasma, saliva, tissue and peripheral blood mononuclear cells (PBMC) of 49 AIDS-KS patients. Overall, 98% of AIDS-KS patients harbored detectable HHV-8. HHV-8 could be detected in 91.6% of KS tissue lesions showing the highest viral load (median log = 3.14 copies/100 ng DNA) followed by saliva and PBMC which were positive in 78%, and 69.2%; respectively. In contrast, HHV-8 was detected in only 37% of plasma samples, which also showed lower viral loads. Men who had sex with men (MSM) were more likely to have three-times higher HHV-8 genome copies in KS lesions when compared with tissues from heterosexuals individuals (OR 3; 95% CI 1.1 to 12.5). These results emphasize the systemic nature of HHV-8-infection and demonstrate the possible role of saliva in HHV-8 transmission among MSM.

The use of selected yeasts for winemaking has clear advantages over the traditional spontaneous fermentation. The aim of this study was to select an indigenous Saccharomyces cerevisiae yeast isolate in order to develop a regional North Patagonian red wine starter culture. A two-step selection protocol developed according to physiological, technological and ecological criteria based on killer interactions was used. Following this methodology, S. cerevisiae isolate MMf9 was selected among 32 indigenous yeasts previously characterized as belonging to different strains according to molecular patterns and killer biotype. This isolate showed interesting technological and qualitative features including high fermentative power and low volatile acidity production, low foam and low sulphide production, as well as relevant ecological characteristics such as resistance to all indigenous and commercial S. cerevisiae killer strains assayed. Red wines with differential volatile profiles and interesting enological features were obtained at laboratory scale by using this selected indigenous strain.

This work describes the application of several analytical techniques to characterize the development of Bordetella pertussis biofilms and to examine, in particular, the contribution of virulence factors in this development. Growth of surface-attached virulent and avirulent B. pertussis strains was monitored in continuous-flow chambers by techniques such as the crystal violet method, and nondestructive methodologies like fluorescence microscopy and Fourier transform (FT) IR spectroscopy. Additionally, B. pertussis virulent and avirulent strains expressing green fluorescent protein were grown adhered to the base of a glass chamber of 1-microm thickness. Three-dimensional images of mature biofilms, acquired by confocal laser scanning microscopy, were quantitatively analysed by means of the computer program COMSTAT. Our results indicate that only the virulent (Bvg(+)) phase of B. pertussis is able to attach to surfaces and develop a mature biofilm. In the virulent phase these bacteria are capable of producing a biofilm consisting of microcolonies of approximately 200 microm in diameter and 24 microm in depth. FTIR spectroscopy allowed us not only to follow the dynamics of biofilm growth through specific biomass and biofilm marker absorption bands, but also to monitor the maturation of the biofilm by means of the increase of the carbohydrate-to-protein ratio.

In this work we evaluate the implantation capacity of the selected S. cerevisiae indigenous strain MMf9 and the quality of the produced wines in a traditional (T) and a modern (M) cellar with different ecological and technological characteristics in North Patagonia (Argentina). Red musts were fermented in 10,000 l vats using the indigenous strain MMf9 as well as the respective controls: a fermentation conducted with a foreign starter culture (BC strain) in M cellar and a natural fermentation in T cellar. Since commercial S. cerevisiae starters are always used for winemaking in M cellar and in order to compare the results, natural fermentations and fermentations conducted by the indigenous strain MMf9 were performed at pilot (200 l) scale in this cellar, concomitantly. Thirty indigenous yeasts were isolated at three stages of fermentation: initial, middle and end. The identification of the yeast biota associated to vinifications was carried out using ITS1-5.8S-ITS2 PCR-RFLP. The intra-specific variability of the S. cerevisiae populations was evaluated using mtDNA-RFLP analysis. Wines obtained from all fermentations were evaluated for their chemical and volatile composition and for their sensory characteristics. A higher capacity of implantation of the indigenous MMf9 strain was evidenced in the fermentation carried out in M cellar (80% at end stage) than the one carried out in T cellar (40%). This behaviour could indicate that each cellar differs in the diversity of S. cerevisiae strains associated to wine fermentations. Moreover a higher capacity of implantation of the native starter MMf9 with regard to the foreign (BC) one was also found in M cellar. The selected indigenous strain MMf9 was able to compete with the yeast biota naturally present in the must. Additionally, a higher rate of sugar consumption and a lower fermentation temperature were observed in vinifications conducted by MMf9 strain with regard to control fermentations, producing wines with favourable characteristics. Even when its implantation in T fermentation was lower than that observed in M one, we can conclude that the wine features from MMf9 fermentations were better than those from their respective controls. Therefore, MMf9 selected indigenous strain could be an interesting yeast starter culture in North Patagonian wines.

Objective: Little is known about the prevalence and distribution of Kaposi's sarcoma-associated herpesvirus (KSHV) infection in the Caribbean. The aim of this study was to determine rates of KSHV seropositivity in various populations in Cuba., Methods: During the years 1998 to 2002 we screened serum samples from 410 subjects in Cuba. Serologic screening for KSHV antibodies was a two-step process using (1) indirect immunofluorescence assay (IFA) specifically reactive to the KSHV latency-associated nuclear antigen (LANA) encoded by open reading frame 73 (ORF73), and (2) confirmatory immunoblot using recombinant KSHV ORF65.2, a lytically expressed, 20-kilodalton protein as the target antigen. Five different populations were studied: (1) 45 AIDS patients with Kaposi's sarcoma (AIDS-KS), (2) 154 HIV-1-infected patients without clinical evidence of KS, (3) 171 HIV-negative blood donors, (4) 27 consecutive kidney transplant recipients, who were HIV-negative, and (5) 13 contacts (sexual contacts or relatives) of the AIDS-KS-affected patients., Results: Among the 45 AIDS-KS subjects, 35 of them (77.8%) were KSHV-seropositive. Thirty-two of the 154 HIV-positive patients without KS (20.8% of them) were KSHV-seropositive, and 6 of the 13 contacts of KS-affected patients (46.2% of them) were infected with KSHV. In contrast to other researchers, we did not find in the populations that we studied in Cuba that KSHV seropositivity was associated with male homosexual or bisexual activity. We found high KSHV seropositivity rates among women reporting sexual contact with bisexual men and among men who had acquired an HIV infection in Africa. There were low rates of KSHV infection among the blood donors (1.2%) and the renal transplant recipients (0.0%). The low rates of KSHV infection that we found among the non-HIV-infected populations in Cuba are similar to patterns found in populations in Europe and in the United States., Conclusions: Together with similar results from Brazil, Jamaica, and the United States of America, our results suggest that KSHV infection is uncommon in some populations in the Western Hemisphere and that KSHV is largely confined to patients with AIDS-associated KS.

The non-treponemic fast detection test of plasmatic reagines (RPR) and the treponemic hemagglutination test of Treponema pallidum were used in the detection of infection due to T. pallidum in 60 males presenting HIV/AIDS infection with clinicoepidemiological diagnosis of syphilis. It was confirmed that 30% presented recently acquired symptomatic or latent syphilis and that 10% had markers of past treated infection or of late acquired latent syphilis, whereas in the other 60% no serological reactivity was detected. A seroprevalence study of reaginic antibodies was conducted by RPR in 59 HIV/AIDS females, using 67 women negative to this virus as controls. All of them had no symptoms compatible with the syphilitic infection. It was concluded that 20.3% and 11.9%, respectively, presented reactivity, which established a probable diagnosis of syphilis or a seroresistance to a previouos syphilis. These results showed a close association between syphilis and HIV/AIDS and that both diseases may coexist in a same patient.

In adult rats, we have induced retinal ischemia and investigated anterogradely labeled surviving retinal ganglion cell (RGC) afferents to the contralateral superior colliculus (SC). The animals received topically in their left eyes two 5-microl drops of saline or saline-containing 0.5% brimonidine (BMD), 1 h before 90 min of retinal ischemia induced by ligature of the left ophthalmic vessels. Two months after ischemia, the anterogradely transported neuronal tracer cholera toxin B subunit (CTB) was injected in the ischemic eyes and animals were processed 4 days later. As controls and for comparison, the retinotectal innervation of unlesioned age-matched control rats was also examined with CTB. In control and experimental animals, serial coronal sections of the mesencephalon and brainstem were immunoreacted for CTB and the area and thickness of the two most superficial layers of the SC containing densely CTB-labeled profiles were estimated with an image analysis system. Ninety minutes of ischemia resulted 2 months later in reduced density of CTB-labeled profiles in the contralateral SC of the vehicle-treated rats, representing less than one half the area occupied by CTB-labeled profiles in control rats. This resulted in shrinkage of these layers and in the presence of areas virtually devoid of CTB immunoreactivity, suggesting orthograde degeneration of retinal terminals and/or decrease of anterograde axonal transport. Topical pretreatment with BMD resulted 2 months later in CTB immunoreactivity that occupied the superficial layers of the contralateral SC in an area of approximately 86% of that observed in the unlesioned control group of animals, indicating that BMD protects against ischemia-induced degeneration of the retinotectal projection, and preserves anterograde axonal transport.

To determine the prevalence rates and serovar distribution of Chlamydia trachomatis cervical infections in Cuban women, two different groups were selected. Group I consisted of 60 human immunodeficiency virus (HIV-1) seropositive women from different regions of Cuba and group II of 60 randomly selected women HIV seronegative and apparently healthy. C. trachomatis was detected in cervical scrapes by mean of nested polymerase chain reaction (PCR) specific for major out membrane protein. The overall prevalence rate of C. trachomatis in cervical scrapes determined by nested PCR was 10% in group I and the estimated prevalence was 6.6% for group II; 83.3% of HIV seropositive women with C. trachomatis infection reported history of pelvic inflammatory disease followed by cervicitis (50%). The control group C. trachomatis-infected women referred a history of cervicitis in 75% of cases. Other reports in the latter group included infertility and pelvic inflamatory disease in 50%. The present study is the first report of C. trachomatis prevalence in Cuba. It showed that there was not significantly difference in the prevalence rate of C. trachomatis between both groups.