Small RNA Differential Expression Analysis Reveals miRNAs Involved in Dormancy Progression in Sweet Cherry Floral Buds.

In sweet cherry (Prunus avium), as in other temperate woody perennials, bud dormancy allows for survival in adverse environmental conditions during winter. During this process, environmental signals such as short days and/or low temperatures trigger internal signals that enable buds to become tolerant to the cold. The process involves tracking chilling units up to chilling the requirement fulfillment to resume growth, a transition involving transcriptional regulation, metabolic signaling, and epigenetic-related regulatory events. Massive sequencing of small RNAs was performed to identify miRNAs involved in sweet cherry dormancy by comparing their expression in field (regular seasonal) and controlled non-stop (continuous) chilling conditions. miRNAs highlighted by sequencing were validated using specific stem-loop PCR quantification, confirming expression patterns for known miRNAs such as miR156e, miR166c, miR172d, miR391, miR482c, and miR535b, as well as for newly proposed miRNAs. In silico prediction of the target genes was used to construct miRNA/target gene nodes. In particular, the involvement of the sweet cherry version for the miR156/SQUAMOSA PROMOTER-BINDING-LIKE PROTEIN genes whose expression was opposite in the two conditions suggests their involvement on dormancy regulation in sweet cherry. miRNA levels indicate that the regulation of stress-related genes and hormone synthesis modulates the expression of calcium metabolism and cell development-associated genes. Understanding the regulatory networks involved in sweet cherry dormancy, particularly in the context of miRNA involvement, represents the first step in the development of new agricultural strategies that may help overcome the increasing challenges presented by global climate change.

Sectorización del trabajo de Médicos Hospitalistas por área geográfica en un hospital universitario con servicios médico quirúrgicos indiferenciados. Proyecto de mejora de calidad / Geographic sectorization of hospitalized patients to health care teams. Impact on teamwork and patient satisfaction

Historically, the wards of hospitals were divided by services such as medicine, surgery and traumatology, among others. To optimize the use of beds, undifferentiated medical surgical services were implemented in different hospitals in the country. This work organization had consequences in several areas, such as teamwork, the sense of belonging, the quality of teaching and travel times, among other factors. In 2018, at a Clinical hospital, we started a quality improvement project that consisted of assigning low complexity internal medicine teams to limited geographic areas, aiming to have sectorized teams. Through some PlanStudy-Do-Act (PDSA) cycles of continuous improvement, more than 80% of the patients were quickly sectorized, however there were multiple threats during the project. Pre- and post-implementation surveys were conducted with nurses, internal medicine residents, and medical Staff, highlighting an improvement in multiple aspects concerning the quality of communication, interdisciplinary work, the time of visits, and satisfaction, among others.

[Geographic sectorization of hospitalized patients to health care teams. Impact on teamwork and patient satisfaction]. / Sectorización del trabajo de Médicos Hospitalistas por área geográfica en un hospital universitario con servicios médico quirúrgicos indiferenciados. Proyecto de mejora de calidad.

Historically, the wards of hospitals were divided by services such as medicine, surgery and traumatology, among others. To optimize the use of beds, undifferentiated medical surgical services were implemented in different hospitals in the country. This work organization had consequences in several areas, such as teamwork, the sense of belonging, the quality of teaching and travel times, among other factors. In 2018, at a Clinical hospital, we started a quality improvement project that consisted of assigning low complexity internal medicine teams to limited geographic areas, aiming to have sectorized teams. Through some PlanStudy-Do-Act (PDSA) cycles of continuous improvement, more than 80% of the patients were quickly sectorized, however there were multiple threats during the project. Pre- and post-implementation surveys were conducted with nurses, internal medicine residents, and medical Staff, highlighting an improvement in multiple aspects concerning the quality of communication, interdisciplinary work, the time of visits, and satisfaction, among others.

Culture-based identification to examine spatiotemporal patterns of fungal communities colonizing wood in ground contact.

Timber durability is often assessed using small wood stakes exposed in direct soil contact, and the assessment generally emphasizes effects on wood rather than organisms involved. Understanding fungal colonization patterns can help identify key decay agents under varying conditions and use these patterns to improve wood protection strategies. Fungal colonization of red alder (Alnus rubra), Douglas-fir (Pseudotsuga menziesii) heartwood/sapwood, and western redcedar (Thuja plicata) field stakes was assessed over 2 y in western Oregon. Spatiotemporal fungal community variations were identified via culturing and DNA sequencing, where 814 isolates were identified from 84 stakes. Forty-six ascomycete genera were identified, with Phialophora, Trichoderma, and Epicoccum species occurring most frequently. Twenty-three basidiomycete genera were identified, with Trametes and Phanerochaete being the most common. Douglas-fir and western redcedar stakes contained the highest and lowest diversity levels, respectively, reflecting natural durability differences of these species. Fungal species abundance was higher below ground than in the above ground and groundline zones, likely reflecting more stable moisture regimes, proximity to soil-based fungi, and potential nutrient migration into wood beneath the soil surface. Ascomycetes were proportionally more abundant early in the exposure period, but basidiomycetes were also observed early in the process, and there appeared to be no consistent colonization pattern.

Nerolidol production in agroinfiltrated tobacco: Impact of protein stability and membrane targeting of strawberry (Fragraria ananassa) NEROLIDOL SYNTHASE1.

The sesquiterpene alcohol nerolidol, synthesized from farnesyl diphosphate (FDP), mediates plant-insect interactions across multiple trophic levels with major implications for pest management in agriculture. We compared nerolidol engineering strategies in tobacco using agroinfiltration to transiently express strawberry (Fragraria ananassa) linalool/nerolidol synthase (FaNES1) either at the endoplasmic reticulum (ER) or in the cytosol as a soluble protein. Using solid phase microextraction and gas chromatography-mass spectrometry (SPME-GCMS), we have determined that FaNES1 directed to the ER via fusion to the transmembrane domain of squalene synthase or hydroxymethylglutaryl - CoA reductase displayed significant improvements in terms of transcript levels, protein accumulation, and volatile production when compared to its cytosolic form. However, the highest levels of nerolidol production were observed when FaNES1 was fused to GFP and expressed in the cytosol. This SPME-GCMS method afforded a limit of detection and quantification of 1.54 and 5.13 pg, respectively. Nerolidol production levels, which ranged from 0.5 to 3.0 µg/g F.W., correlated more strongly to the accumulation of recombinant protein than transcript level, the former being highest in FaNES-GFP transfected plants. These results indicate that while the ER may represent an enriched source of FDP that can be exploited in metabolic engineering, protein accumulation is a better predictor of sesquiterpene production.

Complex interplays between phytosterols and plastid development.

Isoprenoids comprise the largest class of natural compounds and are found in all kinds of organisms. In plants, they participate in both primary and specialized metabolism, playing essential roles in nearly all aspects of growth and development. The enormous diversity of this family of compounds is extensively exploited for biotechnological and biomedical applications as biomaterials, biofuels or drugs. Despite their variety of structures, all isoprenoids derive from the common C5 precursor isopentenyl diphosphate (IPP). Plants synthesize IPP through two different metabolic pathways, the mevalonic acid (MVA) and the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathways that operate in the cytosol-RE and plastids, respectively. MEP-derived isoprenoids include important compounds for chloroplast function and as such, knock-out mutant plants affected in different steps of this pathway display important alterations in plastid structure. These alterations often lead to albino phenotypes and lethality at seedling stage. MVA knock-out mutant plants show, on the contrary, lethal phenotypes already exhibited at the gametophyte or embryo developmental stage. However, the recent characterization of conditional knock-down mutant plants of farnesyl diphosphate synthase (FPS), a central enzyme in cytosolic and mitochondrial isoprenoid biosynthesis, revealed an unexpected role of this pathway in chloroplast development and plastidial isoprenoid metabolism in post-embryonic stages. Upon FPS silencing, chloroplast structure is severely altered, together with a strong reduction in the levels of MEP pathway-derived major end products. This phenotype is associated to misregulation of genes involved in stress responses predominantly belonging to JA and Fe homeostasis pathways. Transcriptomic experiments and analysis of recent literature indicate that sterols are the cause of the observed alterations through an as yet undiscovered mechanism.

Actitudes, prácticas y estilos de vida en adolescentes de instituciones de educación secundaria de la ciudad de popayán, 2016

Objetivo: Determinar los estilos de vida de los entornos escolares, sus conocimientos, actitudes y prácticas hacia su salud, en los estudiantes de educación media vocacional en seis instituciones educativas públicas y privadas de la ciudad de Popayán. Materiales y métodos: Estudio descriptivo, observacional de corte transversal, con muestra probabilística seleccionando 135 estudiantes. Se aplicó un instrumento tipo encuesta para el análisis de factores sociodemográficos, actitudes, prácticas y hábitos de vida de los adolescentes. Resultados: De los estudiantes encuestados son de grado noveno (51,9%); han fumado tabaco alguna vez (38,5%); han consumido alcohol (88,9%); han consumido sustancias psicoactivas (29,6%); realizan actividad física (87,4%); han iniciado su vida sexual (59,3%). Conclusiones: Los hábitos no saludables en los adolescentes se ven influenciados por la vulnerabilidad y entorno social, el uso de la tecnología y la vida moderna, los cuales afectan el tiempo libre y los determinantes sociales en salud.

Objective: To determine the lifestyles of school environments, their knowledge, attitudes and practices towards their health, in vocational secondary education students in six public and private educational institutions from Popayán. Materials and methods: Descriptive, observational cross -sectional study with a probabilistic sample, selecting 135 students, a survey- type instrument was used to analyze the socio-demographic factors, attitudes, practices and life customs of teenagers. Results: The respondent students, they were nineth grade (51.9%); have had smoked cigarettes (38.5%); consumed alcohol (88.9%) and psychoactive substances (29.6%); performed physical activity (87.4%); having started their sexual life (59.3%). Conclusions: Unhealthy habits in adolescents are influenced by vulnerability and social environment, the use of technology and modern life, which affect free time and social determinants in health.

Objetivo: Determinar os estilos de vida no ambiente escolar, seus conhecimentos, atitudes e práticas para sua saúde, nos estudantes do ensino meio em seis escolas públicas e privadas na cidade de Popayán. Métodos: Estudo descritivo, observacional de tipo transversal, com amostra probabilística selecionando 135 estudantes. Aplicou-se o instrumento tipo enquete para as análises dos fatores sociodemogra-ficos, atitudes, práticas e estilos de vida dos adolescentes. Resultados: Os alunos enquistados são do nono ano (51,9%), tem fumado alguma vez (38,5%), tem consumido álcool (88,9%), tem consumido substâncias psicoativas (29,6%), praticam atividade física (87,4%), tem vida sexual ativa (59,3%). Conclusões: os estilos de vida não saludáveis nos adolescentes tem influencia pela vulnerabilidade e entorno social, o uso da tecnologia, a vida moderna os quais afetam seu tempo livre e os determinantes sociais na saúde.

Suppressing Farnesyl Diphosphate Synthase Alters Chloroplast Development and Triggers Sterol-Dependent Induction of Jasmonate- and Fe-Related Responses.

Farnesyl diphosphate synthase (FPS) catalyzes the synthesis of farnesyl diphosphate from isopentenyl diphosphate and dimethylallyl diphosphate. Arabidopsis (Arabidopsis thaliana) contains two genes (FPS1 and FPS2) encoding FPS. Single fps1 and fps2 knockout mutants are phenotypically indistinguishable from wild-type plants, while fps1/fps2 double mutants are embryo lethal. To assess the effect of FPS down-regulation at postembryonic developmental stages, we generated Arabidopsis conditional knockdown mutants expressing artificial microRNAs devised to simultaneously silence both FPS genes. Induction of silencing from germination rapidly caused chlorosis and a strong developmental phenotype that led to seedling lethality. However, silencing of FPS after seed germination resulted in a slight developmental delay only, although leaves and cotyledons continued to show chlorosis and altered chloroplasts. Metabolomic analyses also revealed drastic changes in the profile of sterols, ubiquinones, and plastidial isoprenoids. RNA sequencing and reverse transcription-quantitative polymerase chain reaction transcriptomic analysis showed that a reduction in FPS activity levels triggers the misregulation of genes involved in biotic and abiotic stress responses, the most prominent one being the rapid induction of a set of genes related to the jasmonic acid pathway. Down-regulation of FPS also triggered an iron-deficiency transcriptional response that is consistent with the iron-deficient phenotype observed in FPS-silenced plants. The specific inhibition of the sterol biosynthesis pathway by chemical and genetic blockage mimicked these transcriptional responses, indicating that sterol depletion is the primary cause of the observed alterations. Our results highlight the importance of sterol homeostasis for normal chloroplast development and function and reveal important clues about how isoprenoid and sterol metabolism is integrated within plant physiology and development.

Characterization of Arabidopsis FPS isozymes and FPS gene expression analysis provide insight into the biosynthesis of isoprenoid precursors in seeds.

Arabidopsis thaliana contains two genes encoding farnesyl diphosphate (FPP) synthase (FPS), the prenyl diphoshate synthase that catalyzes the synthesis of FPP from isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). In this study, we provide evidence that the two Arabidopsis short FPS isozymes FPS1S and FPS2 localize to the cytosol. Both enzymes were expressed in E. coli, purified and biochemically characterized. Despite FPS1S and FPS2 share more than 90% amino acid sequence identity, FPS2 was found to be more efficient as a catalyst, more sensitive to the inhibitory effect of NaCl, and more resistant to thermal inactivation than FPS1S. Homology modelling for FPS1S and FPS2 and analysis of the amino acid differences between the two enzymes revealed an increase in surface polarity and a greater capacity to form surface salt bridges of FPS2 compared to FPS1S. These factors most likely account for the enhanced thermostability of FPS2. Expression analysis of FPS::GUS genes in seeds showed that FPS1 and FPS2 display complementary patterns of expression particularly at late stages of seed development, which suggests that Arabidopsis seeds have two spatially segregated sources of FPP. Functional complementation studies of the Arabidopsis fps2 knockout mutant seed phenotypes demonstrated that under normal conditions FPS1S and FPS2 are functionally interchangeable. A putative role for FPS2 in maintaining seed germination capacity under adverse environmental conditions is discussed.