Independent regulation of strigolactones and blumenols during arbuscular mycorrhizal symbiosis in rice.

The apocarotenoid strigolactones (SLs) facilitate pre-symbiotic communication between arbuscular mycorrhizal (AM) fungi and plants. Related blumenol-C-glucosides (blumenols), have also been associated with symbiosis, but the cues that are involved in the regulation of blumenol accumulation during AM symbiosis remain unclear. In rice, our analyses demonstrated a strict correlation between foliar blumenol abundance and intraradical fungal colonisation. More specifically, rice mutants affected at distinct stages of the interaction revealed that fungal cortex invasion was required for foliar blumenol accumulation. Plant phosphate status and D14L hormone signalling had no effect, contrasting their known role in induction of SLs. This a proportion of the SL biosynthetic enzymes, D27 and D17, are equally required for blumenol production. These results importantly clarify that, while there is a partially shared biosynthetic pathway between SL and blumenols, the dedicated induction of the related apocarotenoids occurs in response to cues acting at distinct stages during the root colonisation process. However, we reveal that neither SLs nor blumenols are essential for fungal invasion of rice roots.

Lactobacillus acidophilus LA-5 Ameliorates Inflammation and Alveolar Bone Loss Promoted by A. actinomycetemcomitans and S. gordonii in Mice and Impacts Oral and Gut Microbiomes.

The benefits of probiotics on dysbiotic microbiomes and inflammation are dependent on the tested strain, host factors, and the resident microbiome. There is limited knowledge on the effects of probiotics in A. actinomycetemcomitans-associated periodontitis. Thus, Lactobacillus acidophilus LA5 (LA5) was orally inoculated for 30 days in C57Bl/6 mice infected with A. actinomycetemcomitans JP2 (Aa) and S. gordonii (Sg). Alveolar bone loss, gingival gene expression, and oral and gut microbiomes were determined. LA5 controlled bone loss in Aa+Sg-infected mice, downregulated the expression of Il-1ß and upregulated Il-10 in gingival tissues, and altered the oral and gut microbiomes. LA5 increased the diversity of the oral microbiome of Aa+Sg infected mice, and Aa+Sg and Aa+Sg+LA5 oral or gut microbiomes clustered apart. LA5 induced shifts in Aa+Sg infected mice by increasing the abundance of Muribaculaceae and decreasing Bifidobacteriaceae in the oral cavity and increasing the abundance of Verrucomicrobiae and Eggerthellales in the gut. In conclusion, LA5 oral administration controls experimental Aa-associated periodontitis by altering inflammatory gene expression and the oral and gut microbiomes.

Screening of Saccharomyces cerevisiae metabolite transporters by 13C isotope substrate labeling.

The transportome of Saccharomyces cerevisiae comprises approximately 340 membrane-bound proteins, of which very few are well-characterized. Elucidating transporter proteins' function is essential not only for understanding central cellular processes in metabolite exchange with the external milieu but also for optimizing the production of value-added compounds in microbial cell factories. Here, we describe the application of 13C-labeled stable isotopes and detection by targeted LC-MS/MS as a screening tool for identifying Saccharomyces cerevisiae metabolite transporters. We compare the transport assay's sensitivity, reproducibility, and accuracy in yeast transporter mutant cell lines and Xenopus oocytes. As proof of principle, we analyzed the transport profiles of five yeast amino acid transporters. We first cultured yeast transporter deletion or overexpression mutants on uniformly labeled 13C-glucose and then screened their ability to facilitate the uptake or export of an unlabeled pool of amino acids. Individual transporters were further studied by heterologous expression in Xenopus oocytes, followed by an uptake assay with 13C labeled yeast extract. Uptake assays in Xenopus oocytes showed higher reproducibility and accuracy. Although having lower accuracy, the results from S. cerevisiae indicated the system's potential for initial high-throughput screening for native metabolite transporters. We partially confirmed previously reported substrates for all five amino acid transporters. In addition, we propose broader substrate specificity for two of the transporter proteins. The method presented here demonstrates the application of a comprehensive screening platform for the knowledge expansion of the transporter-substrate relationship for native metabolites in S. cerevisiae.

Caracterização do fluxo ativação da equipa de emergência médica intra-hospitalar / Characterisation of the activation flow of the intra-hospital medical emergency team

As equipas de emergência médica intra-hospitalar têm como principal objetivo a abordagem de situações de deterioração aguda do estado geral do utente. Todos os profissionais devem ter conhecimento da sua existência, das situações que justificam a sua intervenção, bem como do processo de mobilização, desde a ativação até ao destino do final do doente. O objetivo, deste trabalho, é conhecer os contextos do percurso do fluxo de ativação, da equipa médica intra-hospitalar, de uma unidade local de saúde do Norte, desde o processo inicial até ao destino do utente. Como metodologia, foi desenvolvido um estudo de abordagem quantitativa, exploratório, transversal, descritivo, a partir da análise do fluxo de ativações. A amostra é constituída por 78 eventos de ativação, tendo sido elaborado um documento próprio (Instrumento Recolha de Dados) para registo e colheita de dados, para posterior análise. A análise dos resultados, revelou que a amostra é maioritariamente constituída por utentes do sexo feminino (56,4%), com uma média etária de 70,85 anos. O serviço que mais acionou a equipa de emergência médica intra-hospitalar, foi a medicina interna (26,9%), sendo o motivo de ativação, mais frequente, a paragem cardiorrespiratória (28,2%). O principal destino dos utentes foi o serviço de urgência (26,2%) e a sala de emergência (23,1%). O tempo médio de ativação, emerge dos diferentes locais da unidade hospitalar, e segundo os registos do estudo, foi de 6,28 minutos (variável entre 3,82 minutos e 11,7 minutos). Em conclusão defende-se a necessidade de prosseguir os estudos relativamente a estes eventos, para poder, a partir de resultados mais amplos, desenvolver estratégias para diminuir o período de tempo de resposta em determinados serviços mais distantes. Espera-se que os resultados deste estudo possam contribuir para uma maior segurança na prestação de cuidados à pessoa em situação crítica e cultivar o reconhecimento mais precoce de deterioração aguda do doente, em todos os serviços.

In-hospital emergency medical teams have as their main objective the approach of situations of acute deterioration of the general condition of the user. All professionals must be aware of its existence, the situations that justify their intervention, as well as the mobilization process, from activation to the final destination of the patient. The objective of this work is to know the contexts of the activation flow path, of the in-hospital medical team, of a local health unit in the North, from the initial process to the user's destination. As a methodology, a quantitative, exploratory, transversal, descriptive study was developed, based on the analysis of the activation flow. The sample consists of 78 activation events, and a specific document (Data Collection Instrument) was prepared for recording and collecting data for further analysis. The analysis of the results revealed that the sample is mostly made up of female users (56.4%), with an average age of 70.85 years. The service that most called the in-hospital emergency medical team was internal medicine (26.9%), the most frequent reason for activation being cardiorespiratory arrest (28.2%). The main destination of users was the emergency department (26.2%) and the emergency room (23.1%). The average activation time, emerging from the different locations of the hospital unit, and according to the study records, was 6.28 minutes (variable between 3.82 minutes and 11.7 minutes). In conclusion, we defend the need to continue the studies regarding these events, in order to be able, from broader results, to develop strategies to reduce the res- ponse time period in certain more distant services. It is hoped that the results of this study can contribute to greater safety in the provision of care to the person in a critical situation and cultivate earlier recognition of the patient's acute deterioration, in all services.

Hb F Levels in ß-Thalassemia Carriers and Normal Individuals: Known and Unknown Quantitative Trait Loci in the ß-Globin Gene Cluster.

In the already identified quantitative trait loci (QTL), modulating Hb F levels are cis-acting haplotypes of the ß-globin gene cluster itself, although the single nucleotide polymorphisms (SNPs) accounting more for the association, remain uncertain. In this study, the role in Hb F production of previously reported candidate SNPs within the ß-globin gene cluster was reexamined, along with a yet poorly studied variation in the BGLT3 gene. In a sample of ß-thalassemia (ß-thal) carriers, we succeeded in replicating the significant association between increased Hb F levels and rs7482144 (C>T) (HBG2 XmnI), which is the most well-established variation in the cluster influencing the trait. This SNP was found to be in strong linkage disequilibrium (LD) with a variation in the HBBP1 gene [rs10128556 (G>A)], which consistently revealed a similar association signal. Remarkably, much stronger than the latter associations were those involving both rs968857 (T allele) (3' HBBP1) and rs7924684 (G allele) (BGLT3), two SNPs that were also in strong LD. As the pattern of LD detected in the ß-globin gene cluster does not correlate with a tight linkage between markers, complex interactions between SNPs at the cluster seem to modulate Hb F. Seeing that no such associations were detected in normal subjects, the question can be raised on whether, under erythropoiesis stress, epigenetic mechanisms contribute to change the regulation of the entire ß-globin gene cluster. In conclusion, we provide statistical evidence for a new player within the ß-globin gene cluster, BGLT3, that in cooperation with other regions influences Hb F levels in ß-thal carriers.

microRNA390 modulates Nicotiana attenuata's tolerance response to Manduca sexta herbivory.

miR390 is a highly conserved miRNA in plant lineages known to function in growth and development processes, such as lateral root development, and in responses to salt and metal stress. In the ecological model species, Nicotiana attenuata, miR390's biological function remains unknown, which we explore here with a gain-of-function analysis with plants over-expressing (OE-) N. attenuata miR390 (Na-miR390) in glasshouse and natural environments. OEmiR390 plants showed normal developmental processes, including lateral root formation or reproductive output, in plants grown under standard conditions in the glasshouse. OEmiR390 plants did not have dramatically altered interactions with arbuscular mycorrhizal fungi (AMF), Fusarium pathogens, or herbivores. However, Na-miR390 regulated the plant's tolerance of herbivory. Caterpillar feeding elicits the accumulation of a suite of phytohormones, including auxin and jasmonates, which further regulate host-tolerance. The increase in Na-miR390 abundance reduces the accumulation of auxin but does not influence levels of other phytohormones including jasmonates (JA, JA-Ile), salicylic acid (SA), and abscisic acid (ABA). Na-miR390 overexpression reduces reproductive output, quantified as capsule production, when plants are attacked by herbivores. Exogenous auxin treatments of herbivore-attacked plants restored capsule production to wild-type levels. During herbivory, Na-miR390 transcript abundances are increased; its overexpression reduces the abundances of auxin biosynthesizing YUCCA and ARF (mainly ARF4) transcripts during herbivory. Furthermore, the accumulation of auxin-regulated phenolamide secondary metabolites (caffeoylputrescine, dicaffeoylspermidine) is also reduced. In N. attenuata, miR390 functions in modulating tolerance responses of herbivore-attacked plants.

Hipopituitarismo associado a hipoglicemia: relato de 2 casos e revisão da literatura / hypopituitarism associated with hypoglycemia: report of 2 cases and literature review

A ocorrência de hipoglicemia associada a hipopituitarismo é evento raro em pacientes adultos e um pouco mais freqüente em crianças portadoras de disfunção pituitária semelhante. Nos pacientes portadores de insuficiência adrenal, a hipoglicemia é mais prevalente naqueles cuja deficiência de cortisol é secundária à deficiência de ACTH do que naqueles com doença adrenal primária. Isto se deve, provavelmente, ao fato de que, na primeira condição, existem deficiências simultâneas de GH e cortisol. Entretanto, hipoglicemia pode ocorrer nos casos com deficiência isolada de ACTH. Vamos apresentar, inicialmente, dois casos de pacientes adultos, do sexo masculino, com hipopituitarismo e hipoglicemia. A seguir, discutiremos os possíveis mecanismos envolvidos na hipoglicemia, a etiologia mais provável do hipopituitarismo e algumas particularidades presentes nos dois pacientes.