New insights into cadmium tolerance and accumulation in tomato: Dissecting root and shoot responses using cross-genotype grafting.

Cadmium (Cd) is one of the most threatening soil and water contaminants in agricultural settings. In previous studies, we observed that Cd affects the metabolism and physiology of tomato (Solanum lycopersicum) plants even after short-term exposure. The objective of this research was to use cross-genotype grafting to distinguish between root- and shoot-mediated responses of tomato genotypes with contrasting Cd tolerance at the early stages of Cd exposure. This study provides the first report of organ-specific contributions in two tomato genotypes with contrasting Cd tolerance: Solanum lycopersicum cv. Calabash Rouge and Solanum lycopersicum cv. Pusa Ruby (which have been classified and further characterized as sensitive (S) and tolerant (T) to Cd, respectively). Scion S was grafted onto rootstock S (S/S) and rootstock T (S/T), and scion T was grafted onto rootstock T (T/T) and rootstock S (T/S). A 35 µM cadmium chloride (CdCl2) treatment was used for stress induction in a hydroponic system. Both shoot and root contributions to Cd responses were observed, and they varied in a genotype- and/or organ-dependent manner for nutrient concentrations, oxidative stress parameters, antioxidant enzymes, and transporters gene expression. The findings overall provide evidence for the dominant role of the tolerant rootstock system in conferring reduced Cd uptake and accumulation. The lowest leaf Cd concentrations were observed in T/T (215.11 µg g-1 DW) and S/T (235.61 µg g-1 DW). Cadmium-induced decreases in leaf dry weight were observed only in T/S (-8.20%) and S/S (-13.89%), which also were the only graft combinations that showed decreases in chlorophyll content (-3.93% in T/S and -4.05% in S/S). Furthermore, the results show that reciprocal grafting is a fruitful approach for gaining insights into the organ-specific modulation of Cd tolerance and accumulation during the early stages of Cd exposure.

Purple acid phosphatases in coffee-genome-wide identification and the transcriptional profile of selected members in roots upon Pi starvation.

Phosphorus (P) availability is determinant for crop productivity and, despite the sufficient amount of this nutrient in arable land, most of it remains insoluble, leading to the need of high fertilizer input. To cope with P scarcity forecasts and also for cropping costs alleviation, genotypes better adapted to promote P solubilization and absorption are required, especially for perennial crops. Coffee is one of these important perennial crops cultivated in soils with low P availability, and thus we aimed to study adaptive strategies to coffee genotypes in acquire phosphorus. In this study, we focused on rhizosphere phosphatase activity, a major characteristic related to P solubilization from organic pools. To explore the genetic basis of this characteristic, we firstly identified 29 Purple Acid Phosphatases (PAP) genes in C. canephora genome and selected five candidates with higher potential to encode secreted PAPs. We found that C. arabica genotypes have diverse profiles of rhizosphere phosphatase activity, as well as microbial biomass carbon, which we measured to explore the impact of the plant on microbiota and also to discriminate the phosphatase activity origin (plant or microorganism-derived). We selected two C. arabica cultivars with contrasting phosphatase activity and found that one PAP gene has a correlated gene expression profile with this characteristic. This work explores coffee adaptative responses to P starvation conditions, and the information provided can further contribute to breeding programs aiming better adapted genotypes for sustainable agriculture in face of current challenges. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03399-6.

Multiplex CRISPR/Cas9-mediated knockout of the phytoene desaturase gene in Coffea canephora.

Coffea canephora (2n = 2x = 22 chromosomes) is a species with extensive genetic diversity and desirable agronomic traits for coffee breeding programs. However, obtaining a new coffee cultivar through conventional breeding techniques may require more than 30 years of crossing cycles and selection, which hampers the effort of keeping up with market demands and rapidly proposing more resilient to climate change varieties. Although, the application of modern biotechnology tools such as precision genetic engineering technologies may enable a faster cultivar development process. Therefore, we aimed to validate the CRISPR/Cas9 system to generate mutations on a selected genotype of C. canephora, the clone 14. Embryogenic calli and a multiplex binary vector containing two sgRNAs targeting different exons of the CcPDS gene were used. The sgRNAs were under the C. canephora U6 promoter regulation. The target gene encodes phytoene desaturase, an enzyme essential for photosynthesis involved in ß-carotene biosynthesis. Somatic seedlings and embryos with albino, variegated and green phenotypes regenerated after Agrobacterium tumefaciens-mediated genetic transformation were analyzed by verifying the insertion of the Cas9 gene and later by sequencing the sgRNAs target regions in the genome of Robusta modified seedlings. Among them, 77% had the expected mutations, and of which, 50% of them had at least one target with a homozygous mutation. The genotype, temperature of co-cultivation with the bacteria, and light intensity used for subsequent embryo regeneration appeared to strongly influence the successful regeneration of plants with a mutated CcPDS gene in the Coffea genus.

The SAUR gene family in coffee: genome-wide identification and gene expression analysis during somatic embryogenesis.

BACKGROUND: Small auxin-up RNA (SAUR) genes form a wide family supposedly involved in different physiological and developmental processes in plants such as leaf senescence, auxin signaling and transport, hypocotyl development and tolerance to abiotic stresses. The transcription of SAUR genes is quickly induced by auxins, a group of phytohormones of major importance on embryo development. To better understand the distribution and expression profile of such still not explored family in Coffea sp., especially during the development of somatic embryogenesis (SE), SAUR members were characterized in silico using the available Coffea canephora genome data and analyzed for gene expression by RT-qPCR in C. arabica embryogenic samples. METHODS AND RESULTS: Over C. canephora genome 31 CcSAURs were distributed by 11 chromosomes. Out of these 31 gene members, 5 SAURs were selected for gene expression analysis in C. arabica embryogenic materials. CaSAUR12 and CaSAUR18 were the members highly expressed through almost all plant materials. The other genes had more expression in at least one of the developing embryo stages or plantlets. The CaSAUR12 was the only member to exhibit an increased expression in both non-embryogenic calli and the developing embryo stages. CONCLUSION: The identification of SAUR family on C. canephora genome followed by the analysis of gene expression profile across coffee somatic embryogenesis process on C. arabica represents a further additional step towards a better comprehension of molecular components acting on SE. Along with new research about this gene family such knowledge may support studies about clonal propagation methods via somatic embryogenesis to help the scientific community towards improvements into coffee crop.

Genomic and transcriptomic inventory of membrane transporters in coffee: Exploring molecular mechanisms of metabolite accumulation.

The genus Coffea (Rubiaceae) encompasses a group of perennial plant species, including a commodity crop from which seeds are roasted, ground, and infused to make one of the most appreciated beverages in the world. As an important tropical crop restricted to specific regions of the world, coffee production is highly susceptible to the effects of environmental instabilities (i.e., local year-to-year weather fluctuations and global climate change) and threatening pest pressures, not to mention an increasing quality rigor by consumers in industrialized countries. Specialized metabolites are substances that largely affect plant-environment interactions as well as how consumers experience agricultural products. Membrane transporters are key targets, albeit understudied, for understanding and tailoring the spatiotemporal distribution of specialized metabolites as they mediate and control molecular trafficking and substance accumulation. Therefore, we analyzed the transportome of C. canephora encoded within the 25,574 protein-coding genes annotated in the genome of this species and identified 1847 putative membrane transporters. Following, we mined 152 transcriptional profiles of C. canephora and C. arabica and performed a comprehensive co-expression analysis to identify transporters potentially involved in the accumulation of specialized metabolites associated with beverage quality and bioactivity attributes. In toto, this report points to an avenue of possibilities on Coffea genomic and transcriptomic data mining for genetic breeding strategies, which can lead to the development of new, resilient varieties for more sustainable coffee production systems.

Comprehensive characterization of the ALMT and MATE families on Populus trichocarpa and gene co-expression network analysis of its members during aluminium toxicity and phosphate starvation stresses.

Aluminium (Al) toxicity and phosphate deficit on soils are some of the main problems of modern agriculture and are usually associated. Some plants are able to overcome these stresses through exuding organic acids on the rhizosphere, such as citrate and malate, which are exported by MATE (Multi drug and toxin extrusion) and ALMT (Aluminium-activated malate transporter) transporters, respectively. Despite its co-action on acidic soils, few studies explore these two families' correlation, especially on tree crops, therefore we performed a comprehensive description of MATE and ALMT families on Populus trichocarpa as a model species for arboreal plants. We found 20 and 56 putative members of ALMT and MATE families, respectively. Then, a gene co-expression network analysis was performed using broad transcriptomic data to analyze which members of each family were transcriptionally associated. Four independent networks were generated, one of which is composed of members putatively related to phosphate starvation and aluminum toxicity stresses. The PoptrALMT10 and PoptrMATE54 genes were selected from this network for a deeper analysis, which revealed that in roots under phosphate starvation stress the two genes have independent transcriptional profiles, however, on the aluminum toxicity stress they share some common correlations with other genes. The data presented here help on the description of these gene families, of which some members are potentially involved in plant responses to acid soil-related stresses and its exploration is an important step towards using this knowledge on breeding programs for P. trichocarpa and other tree crops.

Evaluating the "satisfaction" and "self-confidence" in nursing students in undergoing simulated clinical experiences.

OBJECTIVE: To evaluate the "satisfaction" and "self-confidence" constructs in nursing students who underwent simulated clinical experiences in semiology and semiotechnique disciplines. METHOD: A descriptive study developed with Nursing students from a federal public university in the interior of Minas Gerais through weekly simulated workshops. The "satisfaction" and "self-confidence" constructs were evaluated by the "Student Satisfaction and Self-Confidence with Learning Scale" and "Simulated Clinical Experiences Satisfaction Scale". RESULTS: The final sample was 45 students. Most were more satisfied (overall average = 4.18) than self-confident (overall average = 4.12). The Spearman correlation test identified a significant and positive association between the "satisfaction" and "self-confidence" constructs. CONCLUSION: Students feel self-confident and satisfied with the use of simulation as an active teaching-learning methodology, highlighting that the higher the satisfaction level, the greater the student's self-confidence.

Evaluating the "satisfaction" and "self-confidence" in nursing students in undergoing simulated clinical experiences / Evaluación de la "satisfacción" y "autoconfianza" en los estudiantes de enfermería que vivieron experiencias clínicas simuladas / Avaliação da "satisfação" e "autoconfiança" em estudantes de enfermagem que vivenciaram experiências clínicas simuladas

Abstract Objective: To evaluate the "satisfaction" and "self-confidence" constructs in nursing students who underwent simulated clinical experiences in semiology and semiotechnique disciplines. Method: A descriptive study developed with Nursing students from a federal public university in the interior of Minas Gerais through weekly simulated workshops. The "satisfaction" and "self-confidence" constructs were evaluated by the "Student Satisfaction and Self-Confidence with Learning Scale" and "Simulated Clinical Experiences Satisfaction Scale". Results: The final sample was 45 students. Most were more satisfied (overall average = 4.18) than self-confident (overall average = 4.12). The Spearman correlation test identified a significant and positive association between the "satisfaction" and "self-confidence" constructs. Conclusion: Students feel self-confident and satisfied with the use of simulation as an active teaching-learning methodology, highlighting that the higher the satisfaction level, the greater the student's self-confidence.

Resumen Objetivo: Evaluar los constructos de "satisfacción" y "autoconfianza" en los estudiantes de enfermería que vivieron experiencias clínicas simuladas en la enseñanza de la semiología y semiotécnica. Método: Se realizó un estudio descriptivo, desarrollado con los estudiantes del curso de enfermería de una universidad pública en el interior de Minas Gerais, a través de talleres simulados sobre una base semanal. Los constructos de "satisfacción" y "autoconfianza" fueron evaluados por la "Escala Satisfacción de los Estudiantes y Autoconfianza con el Aprendizaje" y "Escala de Satisfacción con las Experiencias Clínicas Simuladas". Resultados: La muestra final estuvo compuesta por 45 estudiantes. La mayoría se presentó más satisfecha (media = 4.18) que autoconfiada (media = 4.12). Se identificó, a través de la prueba de correlación de Spearman una asociación significativa y positiva entre los constructos de "satisfacción" y "autoconfianza". Conclusión: Los estudiantes se sientan seguros y satisfechos con el uso de la simulación como una metodología activa del proceso de enseñanza-aprendizaje, destacando que el mayor nivel de satisfacción aumentó la confianza de los estudiantes.

Resumo Objetivo: Avaliar os constructos "satisfação" e "autoconfiança" em estudantes de Enfermagem que vivenciaram experiências clínicas simuladas no ensino de semiologia e semiotécnica. Método: Estudo descritivo, desenvolvido com estudantes do curso de Enfermagem de uma universidade pública federal do interior de Minas Gerais, através de oficinas simuladas semanalmente. Os constructos "satisfação" e "autoconfiança" foram avaliados pela "Escala de Satisfação de Estudantes e Autoconfiança com a Aprendizagem" e "Escala de Satisfação com as Experiências Clínicas Simuladas". Resultados: A amostra final foi de 45 estudantes. A maioria apresentou mais satisfeito (média geral = 4,18) do que autoconfiante (média geral = 4,12). Identificou-se pelo teste de correlação de Spearman uma associação significativa e positiva entre os constructos "satisfação" e "autoconfiança". Conclusão: Os estudantes se sentem autoconfiantes e satisfeitos com o uso da simulação como metodologia ativa de ensino-aprendizagem, destacando-se que quanto maior o nível de satisfação, maior a autoconfiança do aluno.

Molecular characterization of Bacillus thuringiensis strains to control Spodoptera eridania (Cramer) (Lepidoptera: Noctuidae) population

Abstract The main objective of this study was to characterize the toxicity and genetic divergence of 18 Bacillus thuringiensis strains in the biological control of Spodoptera eridania. Bacterial suspensions were added to the S. eridania diet. Half of the selected B. thuringiensis strains caused high mortality seven days after infection. The genetic divergence of B. thuringiensis strains was assessed based on Enterobacterial Repetitive Intergenic Consensus (ERIC) and Repetitive Extragenic Palindromic (REP) sequences, and five phylogenetic groups were formed. Despite their genetic diversity B. thuringiensis strains did not show any correlation between the collection sites and toxicity to larvae. Some B. thuringiensis strains are highly toxic to S. eridania, thus highlighting the potential of their endotoxins as biopesticides.

Genome-wide analysis, transcription factor network approach and gene expression profile of GH3 genes over early somatic embryogenesis in Coffea spp.

BACKGROUND: Coffee production relies on plantations with varieties from Coffea arabica and Coffea canephora species. The first, the most representative in terms of coffee consumption, is mostly propagated by seeds, which leads to management problems regarding the plantations maintenance, harvest and processing of grains. Therefore, an efficient clonal propagation process is required for this species cultivation, which is possible by reaching a scalable and cost-effective somatic embryogenesis protocol. A key process on somatic embryogenesis induction is the auxin homeostasis performed by Gretchen Hagen 3 (GH3) proteins through amino acid conjugation. In this study, the GH3 family members were identified on C. canephora genome, and by performing analysis related to gene and protein structure and transcriptomic profile on embryogenic tissues, we point a GH3 gene as a potential regulator of auxin homeostasis during early somatic embryogenesis in C. arabica plants. RESULTS: We have searched within the published C. canephora genome and found 17 GH3 family members. We checked the conserved domains for GH3 proteins and clustered the members in three main groups according to phylogenetic relationships. We identified amino acids sets in four GH3 proteins that are related to acidic amino acid conjugation to auxin, and using a transcription factor (TF) network approach followed by RT-qPCR we analyzed their possible transcriptional regulators and expression profiles in cells with contrasting embryogenic potential in C. arabica. The CaGH3.15 expression pattern is the most correlated with embryogenic potential and with CaBBM, a C. arabica ortholog of a major somatic embryogenesis regulator. CONCLUSION: Therefore, one out of the GH3 members may be influencing on coffee somatic embryogenesis by auxin conjugation with acidic amino acids, which leads to the phytohormone degradation. It is an indicative that this gene can serve as a molecular marker for coffee cells with embryogenic potential and needs to be further studied on how much determinant it is for this process. This work, together with future studies, can support the improvement of coffee clonal propagation through in vitro derived somatic embryos.

Analysis of gene co-expression networks of phosphate starvation and aluminium toxicity responses in Populus spp.

The adaptation of crops to acid soils is needed for the maintenance of food security in a sustainable way, as decreasing fertilizers use and mechanical interventions in the soil would favor the reduction of agricultural practices' environmental impact. Phosphate deficiency and the presence of reactive aluminum affect vital processes to the plant in this soil, mostly water and nutrient absorption. From this, the understanding of the molecular response to these stresses can foster strategies for genetic improvement, so the aim was to broadly analyze the transcriptional variations in Poupulus spp. in response to these abiotic stresses, as a plant model for woody crops. A co-expression network was constructed among 3,180 genes differentially expressed in aluminum-stressed plants with 34,988 connections. Of this total, 344 genes presented two-fold transcriptional variation and the group of genes associated with those regulated after 246 hours of stress had higher number of connections per gene, with some already characterized genes related to this stress as main hubs. Another co-expression network was made up of 8,380 connections between 550 genes regulated by aluminum stress and phosphate deficiency, in which 380 genes had similar profile in both stresses and only eight with transcriptional variation higher than 20%. All the transcriptomic data are presented here with functional enrichment and homology comparisons with already characterized genes in another species that are related to the explored stresses, in order to provide a broad analysis of the co-opted responses for both the stresses as well as some specificity. This approach improves our understanding regarding the plants adaptation to acid soils and may contribute to strategies of crop genetic improvement for this condition that is widely present in regions of high agricultural activity.

Vivência do homem diante do adoecimento pelo câncer: implicações para o cuidado em saúde / La experiencia del hombre frente a la enfermedad por cáncer: implicaciones para la atención en salud / Man's experience facing cancer illness: implications for health care

Objetivo: compreender como o homem vivencia o processo de adoecimento do câncer, desde o diagnóstico até o tratamento. Método: estudo descritivo, exploratório e qualitativo, realizado com dez homens em tratamento oncológico, em um município do interior de Minas Gerais, entre março e maio de 2017. Os dados foram coletados a partir de entrevistas gravadas, posteriormente transcritos na íntegra e submetidos a análise temática do conteúdo. Resultados: emergiram duas categorias: ''A descoberta do diagnóstico, o tratamento e as mudanças no cotidiano: enfrentamentos e ressignificações'' e ''O cuidado oferecido pela equipe de saúde''. Conclusão: a vivência de uma doença oncológica é permeada por negação, raiva, angústia, tristeza e temor. O homem se depara com possibilidade de morte, ameaça ao seu futuro e dificuldades para se adaptar à nova vida. A família, amigos e os profissionais de saúde são importantes para o controle e o enfrentamento da doença.

Objective: to understand how men experience the process of becoming ill with cancer, from diagnosis to treatment. Method: a descriptive, exploratory and qualitative study, conducted with ten men undergoing cancer treatment, in a municipality in the interior of Minas Gerais, between March and May 2017. Data were collected from recorded interviews, later transcribed in full and submitted to thematic analysis of the content. Results: two categories emerged: "The discovery of diagnosis, the treatment and the changes in daily life: coping and resignification'' and; ''The care offered by the health team''. Conclusion: the experience of an oncological disease is permeated by denial, anger, anguish, sadness and fear. The man faces the possibility of death, threat to his future and has difficulty adapting to his new life. Family, friends and health professionals are important for the control and coping with the disease.

Objetivo: comprender cómo los hombres experimentan el proceso de la enfermedad del cáncer, desde el diagnóstico hasta el tratamiento. Método: estudio descriptivo, exploratorio y cualitativo, realizado con diez hombres sometidos a tratamiento, en un municipio del interior de Minas Gerais, entre marzo y mayo de 2017. Los datos fueron recolectados a partir de entrevistas grabadas, transcritas en su totalidad y sometidas al Análisis Temático de Contenido. Resultados: emergieron dos categorías: "El descubrimiento del diagnóstico, el tratamiento y los cambios en la vida cotidiana: afrontamiento y nuevas significaciones" y; "La atención ofrecida por el equipo de salud". Conclusión: la experiencia de una enfermedad oncológica está impregnada de negación, ira, angustia, tristeza y miedo. El hombre enfrenta la posibilidad de la muerte, amenaza a su futuro y tiene dificultades para adaptarse a su nueva vida. Familia, amigos y profesionales de salud son importantes para controlar y hacer frente a la enfermedad.

Da decisão à vivência da cesariana: a perspectiva da mulher / From decision to cesarian: the woman perspective / De la decisión a la vivencia de la cesárea: la perspectiva de la mujer

OBJETIVO: descrever o processo de decisão da mulher primípara pela via de nascimento, compreendendo a vivência da cesariana pela mesma. MÉTODO: trata-se de uma pesquisa do tipo descritiva, com abordagem qualitativa. As participantes foram dez puérperas primíparas que vivenciaram a cesariana. Na coleta de dados, utilizou-se a técnica de entrevista aberta com posterior análise temática, aplicando a técnica Laurence Bardin. RESULTADOS: Foram levantadas duas categorias: A decisão pela via de nascimento e o momento da cesariana. Na primeira, foi possível perceber que a maioria das mulheres já chega ao consultório decidida sobre a via de nascimento, no entanto, nem todas conseguiram prosseguir com a escolha inicial. Na segunda, retrata-se como foi vivenciar a cesariana, estando a mulher amparada pelos entes queridos e apoiada pelos profissionais de saúde. CONCLUSÃO: Ressalta-se a importância de a equipe de saúde atuar efetivamente no compartilhamento de informações e na construção do vínculo, desde o pré-natal até o puerpério, em que a mulher exerce o real protagonismo do parto

OBJECTIVE: to describe the primiparous woman's decision process for the birth way, understanding the cesarean experience by the same woman. METHOD: This is a descriptive research with a qualitative approach. The participants were ten primiparous women who underwent cesarean section. In the data collection, the technique of open interview with thematic analysis, applying the Laurence Bardin's technique. RESULTS: There were two categories: the decision by birth and the time of cesarean section. In the first decision, it was possible to notice that most of the women already arrive at the office decided about the birth way, however, not all of them were able to proceed with the initial choice. The second one portrays how it was to experience the cesarean section, the woman supported by their loved ones and supported by health professionals. CONCLUSION: It is important to emphasize the importance of the health team to act effectively in information sharing and in the bond building, from the prenatal to the puerperium, in which the woman performs the real role of childbirth

OBJETIVO: describir el proceso de decisión de la mujer primípara por la vía de nacimiento y comprender la vivencia de la cesárea por la misma. MÉTODO: se trata de una investigación del tipo descriptivo, con abordaje cualitativo. Las participantes fueron diez mujeres que hubieran dado a luz primíparas que experimentaron la cesárea. En la recolección de datos, se utilizó la técnica de entrevista abierta con posterior análisis temático utilizando la técnica Laurence Bardin. RESULTADOS: Se levantaron dos categorías: La decisión por la vía de nacimiento y el momento de la cesárea. En la primera, fue posible percibir que la mayoría de las mujeres ya llega al consultorio decidida sobre la vía de nacimiento, sin embargo, no todas han logrado proseguir con la elección inicial. La segunda retrata cómo fue vivir la cesárea, estando la mujer amparada por los seres queridos y apoyados por los profesionales de la salud. CONCLUSIÓN: Se resalta la importancia del equipo de salud actuar efectivamente en el intercambio de informaciones y en la construcción del vínculo, desde el prenatal hasta el puerperio, en que la mujer ejerce el real protagonismo del parto

Gene expression in two contrasting hybrid clones of Eucalyptus camaldulensis x Eucalyptus urophylla grown under water deficit conditions.

The physiological and molecular responses to water stress are mediated by a range of mechanisms, many of which involve abscisic acid (ABA)-dependent signaling pathways. In addition, plants contain drought response genes that can be induced by ABA-independent routes, mediated by secondary messengers such as Ca2+, or regulated by epigenetic modifications. The complex processes involved in the response to water stress can be investigated using molecular techniques to evaluate the expression patterns of genes of interest and to infer the behavior of different genotypes and species. In the present study, we first analyzed the stability of a set of reference genes for normalization of the gene expression with real-time quantitative polymerase chain reaction (RT-qPCR), since there were no results related to the genotype used in this study. We verified that although there were some variations between algorithms used, the three most stable reference genes were SAND, PP2A-3 and EF-1α. The expressions of genes encoding for proteins associated with drought-tolerance responses, namely 9-cis-epoxycarotenoid dioxygenase 3 (EgrNCED3), pyrabactin resistance 1 (EgrPYR1), dehydration-responsive element-binding 2.5 (EgrDREB2.5) transcription factors, calcium-dependent protein kinase 26 (EgrCDPK26), methyl transferase 1 (EgrMET1) and deficient in DNA methylation 1 (EgrDDM1) protein, were determined by RT-qPCR in leaf samples from drought sensitive (VM05) and drought tolerant (VM01) clones of the hybrid Eucalyptus camaldulensis x Eucalyptus urophylla grown under water stress and irrigation conditions. When the two clones were maintained under conditions of water deficiency, VM01 exhibited higher expression levels of EgrNCED3 and EgrPYR1 genes than VM05 at all sampling times, implying that ABA biosynthesis and subsequent induction of the ABA-dependent cascade mediated by the PYR1-ABA receptor complex were enhanced in the tolerant clone. Under water-stress conditions, this clone also presented increased expression of the EgrDREB2.5 gene, representative of an ABA-independent cascade, and of the EgrCPK26 gene, related to stomatal opening and closure. On the other hand, the expression levels of EgrMET1 and EgrDDM1 genes in the sensitive clone were higher than in the tolerant clone under all conditions, showing a putative impact of epigenetic modifications on tolerance to water deficiency. The results obtained indicate that the superior ability of the VM01-tolerant clone to perceive water deficiency and activate drought-resistance genes is associated with the high expression levels of EgrNCED3, EgrPYR1 and EgrDREB2.5 under water-stress conditions. These findings will facilitate future research on the functional characterization of stress-related response genes, the identification of molecular markers, the evaluation of drought tolerance and genetic transformation in tree species.

Gene Expression Profile Analysis is Directly Affected by the Selected Reference Gene: The Case of Leaf-Cutting Atta Sexdens.

Although several ant species are important targets for the development of molecular control strategies, only a few studies focus on identifying and validating reference genes for quantitative reverse transcription polymerase chain reaction (RT-qPCR) data normalization. We provide here an extensive study to identify and validate suitable reference genes for gene expression analysis in the ant Atta sexdens, a threatening agricultural pest in South America. The optimal number of reference genes varies according to each sample and the result generated by RefFinder differed about which is the most suitable reference gene. Results suggest that the RPS16, NADH and SDHB genes were the best reference genes in the sample pool according to stability values. The SNF7 gene expression pattern was stable in all evaluated sample set. In contrast, when using less stable reference genes for normalization a large variability in SNF7 gene expression was recorded. There is no universal reference gene suitable for all conditions under analysis, since these genes can also participate in different cellular functions, thus requiring a systematic validation of possible reference genes for each specific condition. The choice of reference genes on SNF7 gene normalization confirmed that unstable reference genes might drastically change the expression profile analysis of target candidate genes.

Morpho and Cytological Differentiation of Calli of Eucalyptus grandis x Eucalyptus urophylla During Somatic Embryogenesis

ABSTRACT The aim of this study was to induce and analyze embryogenic calli from two types of explants (leaves and meristems) of the hybrid Eucalyptus grandis x Eucalyptus urophylla. Leaves and meristems of plants kept in a nursery were disinfected and inoculated in Petri dishes containing MS culture medium supplemented with different concentrations of the growth regulator dicamba (1.13, 4.52, and 9.04 µM) and without it. At 60 days of culturing, the calli were analyzed by scanning electron microscopy and at 90 days were evaluated by light microscopy in regard to the embryogenic characteristics of the cells. Different type of calli were induced in leaf explants, designated as Type I with light yellow coloring, Type II with dark yellow coloring, and Type III of brown coloring; however, only Type I had embryogenic characteristics. In the meristematic explants, only one type of callus was induced, and it had embryogenic characteristics. At 90 days of culturing, the formation of somatic embryos in the different embryogenic stages was observed and the formation of procambium, protoderm, and ground meristem tissues. At 150 days of culturing, the concentration of 1.13 µM of dicamba was prominent in the formation of somatic embryos in the different embryogenic stages.

O uso da simulação no ensino da técnica de aspiração de vias aéreas: ensaio clínico randomizado controlado / Use of simulations in the teaching of the airway aspiration technique: controlled randomized clinical trial / El uso de la simulación en la enseñanza de la técnica de aspiración de vias aéreas: ensayo clínico randomizado

Estudo com o objetivo de avaliar o efeito do uso da simulação no ensino da habilidade de aspiração de vias aéreas inferiores. Trata-se de ensaio clínico controlado randomizado realizado em uma universidade pública federal em outubro de 2016. Amostra de 33 estudantes alocados mediante sorteio aleatório nos grupos intervenção (n=17) e controle (n=16). A intervenção avaliada foi o ensino da técnica de aspiração de vias aéreas inferiores utilizando a simulação (oficinas individuais e debriefing), e o controle foi o ensino da técnica pelo método tradicional (aula expositiva e treinamento em grupo). Os desfechos avaliados foram o conhecimento teórico e prático sobre a habilidade ensinada, cujas pontuações máximas eram, respectivamente, 19 e 29 pontos. Os dados foram analisados utilizando-se estatística descritiva e inferencial. No conhecimento teórico, a média de acertos do grupo intervenção foi de 15,1 pontos e no grupo-controle foi de 15,5 pontos. Na avaliação prática, a pontuação média no grupo intervenção foi de 22,9 pontos e no grupo-controle foi de 20,8 pontos. Não houve diferença na média de acertos no conhecimento teórico e prático sobre a habilidade ensinada entre os grupos intervenção e controle. Entretanto, alunos que participaram das atividades simuladas relataram mais confiança e segurança no desenvolvimento da técnica em laboratório. Sugere-se a replicação do estudo com amostra maior para comparação dos achados. Registro Brasileiro de Ensaios Clínicos: RBR-8bsmyz.

This study aimed to evaluate the effect of the use of simulations to teach lower airway aspiration. This is a randomized controlled clinical trial conducted at a federal public university in October 2016. The sample was composed of 33 students randomly assigned to intervention (n = 17) and control (n = 16) groups. The intervention evaluated the teaching of the lower airway aspiration technique using simulations (individual workshops and debriefing), and the control situation was represented by the teaching of the technique through the traditional method (expository class and group training). The outcomes evaluated were the theoretical and practical knowledge about the skill taught, whose maximum scores were, respectively, 19 and 29 points. Data were analyzed using descriptive and inferential statistics. In the theoretical knowledge, the average number of correct answers in the intervention group was 15.1 points and in the control group it was 15.5 points. In the practical evaluation, the mean score in the intervention group was 22.9 points and in the control group it was 20.8 points. There was no difference in the average of correct theoretical and practical knowledge about the ability taught between the intervention and control groups. However, students who participated in the simulated activities reported more confidence and reliability in the development of the technique in laboratory. We suggest the replication of the study with a larger sample for comparison of the findings. Brazilian Registry of Clinical Trials: RBR-8bsmyz.

Subspecies and diet affect the expression of genes involved in lipid metabolism and chemical composition of muscle in beef cattle.

Fourteen Nellore and 14 Angus young bulls with BW of 381±11.8kg were randomly assigned into 2 feeding groups (whole shelled corn without forage (WSC) or corn silage and ground corn (GC)) to evaluate chemical composition and expression of genes involved in lipid metabolism in the longissimus thoracis (LT). We hypothesized that bulls fed the WSC diet have greater amounts of intramuscular fat and Angus have higher expression levels of PPAR and SREBF. Meat from Angus bulls had greater ether extract compared to Nellore (P<0.05). Muscle from bulls fed the WSC diet had greater expression of PPARA (P<0.05) and lower levels of SREBF1 expression (P<0.01). The LT of Nellore fed GC had greater expression of FABP4, ACACA and SCD genes (P<0.01). In conclusion, the greater concentration of starch in the WSC diet did not increase marbling in the beef of bulls fed this diet due to the reduced expression of SREBF1.

Molecular epidemiology of Streptococcus agalactiae isolated from mastitis in Brazilian dairy herds

Abstract Streptococcus agalactiae is one of the most common pathogens leading to mastitis in dairy herds worldwide; consequently, the pathogen causes major economic losses for affected farmers. In this study, multilocus sequence typing (MLST), genotypic capsular typing by multiplex polymerase chain reaction (PCR), and virulence gene detection were performed to address the molecular epidemiology of 59 bovine (mastitis) S. agalactiae isolates from 36 dairy farms located in the largest milk-producing mesoregions in Brazil (Minas Gerais, São Paulo, Paraná, and Pernambuco). We screened for the virulence genes bac, bca, bibA, cfb, hylB, fbsA, fbsB, PI-1, PI-2a, and PI-2b, which are associated with adhesion, invasion, tissue damage, and/or immune evasion. Furthermore, five capsular types were identified (Ia, Ib, II, III, and IV), and a few isolates were classified as non-typeable (NT). MLST revealed the following eight sequence types (STs): ST-61, ST-67, ST-103, ST-146, ST-226, ST-314, and ST-570, which were clustered in five clonal complexes (CC64, CC67, CC103, CC17, and CC314), and one singleton, ST-91. Among the virulence genes screened in this study, PI-2b, fbsB, cfb, and hylB appear to be the most important during mastitis development in cattle. Collectively, these results establish the molecular epidemiology of S. agalactiae isolated from cows in Brazilian herds. We believe that the data presented here provide a foundation for future research aimed at developing and implementing new preventative and treatment options for mastitis caused by S. agalactiae.

Molecular epidemiology of Streptococcus agalactiae isolated from mastitis in Brazilian dairy herds.

Streptococcus agalactiae is one of the most common pathogens leading to mastitis in dairy herds worldwide; consequently, the pathogen causes major economic losses for affected farmers. In this study, multilocus sequence typing (MLST), genotypic capsular typing by multiplex polymerase chain reaction (PCR), and virulence gene detection were performed to address the molecular epidemiology of 59 bovine (mastitis) S. agalactiae isolates from 36 dairy farms located in the largest milk-producing mesoregions in Brazil (Minas Gerais, São Paulo, Paraná, and Pernambuco). We screened for the virulence genes bac, bca, bibA, cfb, hylB, fbsA, fbsB, PI-1, PI-2a, and PI-2b, which are associated with adhesion, invasion, tissue damage, and/or immune evasion. Furthermore, five capsular types were identified (Ia, Ib, II, III, and IV), and a few isolates were classified as non-typeable (NT). MLST revealed the following eight sequence types (STs): ST-61, ST-67, ST-103, ST-146, ST-226, ST-314, and ST-570, which were clustered in five clonal complexes (CC64, CC67, CC103, CC17, and CC314), and one singleton, ST-91. Among the virulence genes screened in this study, PI-2b, fbsB, cfb, and hylB appear to be the most important during mastitis development in cattle. Collectively, these results establish the molecular epidemiology of S. agalactiae isolated from cows in Brazilian herds. We believe that the data presented here provide a foundation for future research aimed at developing and implementing new preventative and treatment options for mastitis caused by S. agalactiae.