RESUMEN
Grapevine (Vitis vinifera L.) crops are continuously exposed to biotic and abiotic stresses, which can cause genetic and epigenetic alterations. To determine the possible effects of grapevine cryopreservation on the regulation of DNA demethylase genes, this work studied the expression of DNA demethylase genes in cryopreserved and post-cryopreserved grapevine tissues. V. vinifera DNA demethylases were characterized by in silico analysis, and gene expression quantification was conducted by RTâqPCR. Three DNA demethylase sequences were found: VIT_13s0074g00450 (VvDMT), VIT_08s0007g03920 (VvROS1), and VIT_06s0061g01270 (VvDML3). Phylogenetic analysis revealed that the sequences from V. vinifera and A. thaliana had a common ancestry. In the promoters of responsive elements to transcription factors such as AP-2, Myb, bZIP, TBP, and GATA, the conserved domains RRM DME and Perm CXXC were detected. These responsive elements play roles in the response to abiotic stress and the regulation of cell growth. These data helped us characterize the V. vinifera DNA demethylase genes. Gene expression analysis indicated that plant vitrification solution 2 (PVS2) treatment does not alter the expression of DNA demethylase genes. The expression levels of VvDMT and VvROS1 increased in response to cryopreservation by vitrification. Furthermore, in post-cryopreservation, VvROS1 was highly induced, and VvDML3 was repressed in all the treatment groups. Gene expression differences between different treatments and tissues may play roles in controlling methylation patterns during gene regulation in tissues stressed by cryopreservation procedures and in the post-cryopreservation period during plant growth and development.
Asunto(s)
Criopreservación , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas , Vitis , Vitis/genética , Vitis/crecimiento & desarrollo , Criopreservación/métodos , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Filogenia , Semillas/genética , Semillas/crecimiento & desarrollo , Desmetilación del ADN , Cigoto/metabolismo , Metilación de ADN , Crioprotectores/farmacologíaRESUMEN
RESUMEN Objetivo. Determinar la seroprevalencia de brucelosis y leptospirosis en cerdos de granjas de pequeña escala en áreas rurales del partido de Tandil, Argentina; analizar los factores de riesgo asociados; describir los serogrupos de Leptospira spp. prevalentes y determinar la distribución espacial de ambas enfermedades. Materiales y métodos. Se obtuvieron 340 muestras de suero. Se utilizaron la prueba de aglutinación en placa, la prueba de Rosa de Bengala y el Test de Polarización de la Fluorescencia para detectar anticuerpos contra Brucella spp. Se aplicó la Prueba de Aglutinación Microscópica para el diagnóstico de leptospirosis. Se utilizaron métodos estadísticos para evaluar los factores asociados a las infecciones. Resultados. Se observaron seroprevalencias de 0 y 22,6% para brucelosis y leptospirosis, respectivamente. Los serogrupos de Leptospira más prevalentes fueron: Canicola, Ballum, Icterohaemorrhagiae y Pomona. Los principales factores de riesgo asociados a la infección por leptospirosis fueron el mayor número de cerdos, la presencia de equinos y la presencia de jabalíes cerca o dentro de las granjas. Se asoció el uso de subproductos de cereal como alimento para los animales y el uso de cajón para el destete. Se detectó un agrupamiento espacial significativo de seropositividad a Leptospira spp. en un área de baja altitud. Conclusiones. La brucelosis probablemente esté controlada, pero Leptospira spp. se encuentra presente en estas producciones. El conocimiento de la seroprevalencia de la brucelosis y la leptospirosis, su distribución espacial y los factores de riesgo asociados puede ser útil para la prevención y el control de enfermedades zoonóticas endémicas en la región.
ABSTRACT Objectives. To determine the seroprevalence of brucellosis and leptospirosis in pigs reared on small-scale farms in rural areas of the district of Tandil, Buenos Aires Province, Argentina; to analyse the associated risk factors; to describe the prevalent Leptospira spp. serogroups; and to determine the spatial distribution of both diseases. Materials and methods. A total of 340 serum samples were obtained. Buffer Plate Agglutination Test, Rose Bengal Test, and Fluorescence Polarisation Assay were used to detect antibodies against Brucella spp. Microscopic Agglutination Test was applied for serological diagnosis of leptospirosis. Statistical methods were used to evaluate the factors associated with the infections. Results. Seroprevalence of 0% and 22.6% was observed for brucellosis and leptospirosis, respectively. The most prevalent Leptospira serogroups identified were Canicola, Ballum, Icterohaemorrhagiae and Pomona. The main risk factors associated with leptospirosis infection were farms with a higher number of pigs and the presence of horses and wild boars near or within the farms. The use of milling by-products of cereal grains as animal feed and the presence of weaning cages were also associated. A relevant spatial cluster of seropositivity to Leptospira spp. was identified in a low-altitude area. Conclusions. Our results suggest that brucellosis is probably controlled, but Leptospira spp. are present in the farming systems under study. Knowledge of brucellosis and leptospirosis seroprevalence, spatial distribution and associated risk factors can be useful for the prevention and control of endemic zoonotic diseases in the region.
RESUMEN
Parasite M17 leucine aminopeptidases (LAPs) have been associated with critical roles in different key functions such as the nutrition, migration, and invasion of the natural host. Native or recombinant LAP used as a vaccine antigen has proved effective to elicit protection against Fasciola hepatica infection in sheep, pointing to potential vaccine candidates against fascioliasis in ruminant species. Previously, the FhLAP1, abundantly secreted in vitro by the mature adult parasite was used as a vaccine antigen obtaining promising protection results against F. hepatica challenge in small ruminants. Here, we report the biochemical characterization of a second recombinant LAP (FhLAP2) associated with the juvenile stage of F. hepatica. FhLAP2 showed aminopeptidase activity using different synthetic substrates, including leucine, arginine, and methionine and was increased in the presence of Mn+ 2 and Mg+ 2. The activity was inhibited by bestatin, 1,10-phenanthroline, and EDTA, specific inhibitors of aminopeptidase and/or metalloproteases. Finally, the recombinant FhLAP2 functional form was tested in combination with Freund's incomplete adjuvant in an immunization trial in mice followed by an experimental challenge with F. hepatica metacercariae. The immunization with FhLAP2/FIA resulted in a significant reduction of parasite recovery compared to control groups. The immunized group elicited total specific IgG and subclasses IgG1 and IgG2 antibody responses. This study highlights the potential of a new candidate vaccine formulation with potential applications in natural ruminant hosts, especially those targeting the juvenile stage.
Asunto(s)
Fasciola hepatica , Fascioliasis , Enfermedades de las Ovejas , Vacunas , Ovinos , Ratones , Animales , Fascioliasis/prevención & control , Fascioliasis/veterinaria , Leucil Aminopeptidasa/química , Leucina , Anticuerpos Antihelmínticos , Enfermedades de las Ovejas/parasitologíaRESUMEN
Cryopreservation has the potential for long-term germplasm storage. The metabolic pathways and gene regulation involved in cryopreservation procedures are still not well documented. Hence, the genetic expression profile was evaluated using RNA-Seq in zygotic embryos of grapevines subjected to cryopreservation by vitrification. Sequencing was performed on the Illumina NextSeq 500. The average alignment of reads was 96% against the reference genome. The expression profiles showed 229 genes differentially expressed (186 repressed and 46 induced). The main biological processes showing upregulated enrichment were related to nucleosome assembly, while downregulated processes were related to organ growth. The most highly repressed processes were associated with the organization of the cell wall and membrane components. The unnamed protein product and 17.3 kDa class II heat shock protein (HSP) were highly induced, while ATPase subunit 1 and expansin-A1 were repressed. The response to the cooling and warming process during cryopreservation probably indicates that the changes occurring in transcription may be related to epigenetics. In addition, the cell exhibits an increase in the reserve of nutrients while seeking to survive modestly using available energy and pausing the plant's development. Additionally, energy containment occurred to cope with the stress caused by the treatment where deactivation of components of the cell membrane was observed, possibly due to changes in fluidity caused by alterations in temperature.
Asunto(s)
Criopreservación , Transcriptoma , Criopreservación/métodos , Frío , Vitrificación , Transición de FaseRESUMEN
RESUMO Este artigo parte da compreensão de interdisciplinaridade como modo de operar face às visões fragmentadas presentes nos processos de produção e de socialização do conhecimento. Objetiva compartilhar reflexões que problematizam a interdisciplinaridade a partir da experiência de formação acadêmica na pós-graduação de um grupo de pesquisa cujos integrantes expressam diversidade de formação e inserção profissional e se debruçam sobre relações entre políticas públicas, saúde e necessidades das pessoas. As reflexões foram elaboradas com base em questões emergentes nos encontros sistemáticos do grupo, que foi tomado como estratégia teórico-metodológica, e sustentadas a partir do diálogo entre saúde coletiva, como campo de saber e de prática, e psicologia social da práxis, formulada por Enrique Pichon-Rivière. O eixo articulador dessa experiência é a formação em sentido amplo que se manifesta pelo princípio de indissociabilidade ensino-pesquisa-extensão, i.e., aprender-investigar-fazer, e como autoformação em um processo mútuo e de ação reflexiva, de aprender a aprender. Nesse sentido, argumenta-se que é metaformação e só pode se dar na perspectiva de diálogos de saberes e interdisciplinares.
ABSTRACT This article is based on the understanding of interdisciplinarity as a way to act against the fragmented visions present in the processes of knowledge production and socialization. The objective is to share reflections that problematize interdisciplinarity from the experience of graduate academic training in a research group whose members have diverse training and professional fields and focus on the relationships between public policy, health and human needs. The reflections were based on issues that emerged during the group's systematic meetings, and the group was understood as a theoretical and methodological strategy, and was sustained by the dialogue between collective health as a field of knowledge and practice, and the social psychology of praxis, formulated by Enrique Pichon-Rivière. The articulating axis of this experience is training in its broadest sense, manifested on the principle of the inseparability of (i.e., learning-research-doing) and as self-training in a mutual process of reflective teaching-research-outreach action, of learning how to learn. In this sense, it is argued that it is a metatraining that can only take place from the perspective of knowledge dialogues and interdisciplinarity.
RESUMEN
RESUMO Pretendeu-se refletir sobre a relevância de contribuições feministas como referenciais contrahegemônicos para uma análise crítica de práticas de saúde desenvolvidas em um serviço da atenção básica do Sistema Único de Saúde. Observaram-se ações de profissionais em uma Unidade Básica de Saúde com o objetivo de analisar como se manifestam questões de gênero na atenção básica a partir de perspectiva da psicologia social postulada por Pichon-Rivière em diálogo com produções feministas. A partir de discussão crítica perante visões dicotômicas, como público e privado, e discursos naturalistas, foram identificados três aspectos relevantes: divisão sexual do trabalho, maternidade e ausência paterna; centralidade das práticas de saúde da mulher na reprodução; e binarismo das políticas públicas de saúde. Isso permitiu identificar aspectos importantes acerca das práticas de saúde na atenção básica. Espera-se que este estudo, ao examinar a naturalização e a reprodução de sexismo, racismo e classismo em meio às práticas consideradas, fomente discussões futuras no campo da saúde que caminhem no sentido de uma produção de saberes e práticas comprometidos com a contestação de tais desigualdades.
ABSTRACT It was intended to reflect on the relevance of feminist contributions as counter-hegemonic references for a critical analysis of health practices developed in a primary care service of the Unified Health System. The actions of professionals in a Basic Health Unit were observed with the aim of analyzing how gender issues are manifested in primary care from the perspective of social psychology postulated by Pichon-Rivière in dialogue with feminist productions. From a critical discussion on dichotomous views, such as public and private, and naturalistic discourses, three relevant aspects were identified: sexual division of work, maternity and paternal absence; centrality of women's health practices in reproduction; and binarism of public health policies. This allowed us to identify important aspects about health practices in primary care. It is hoped that this study, by examining the naturalization and reproduction of sexism, racism, and classism among the practices considered, will encourage future discussions in the field of health that move towards a production of knowledge and practices committed to contesting such inequalities.