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Extracellular vesicles (EVs) are nanoparticles that are released by cells and participate in the transfer of information. It is now known that EVs from mammalian cells are involved in different physiological and pathophysiological processes (antigen presentation, tissue regeneration, cancer, inflammation, diabetes, etc.). In the past few years, several studies on plants have demonstrated that EVs are also key tools for plant intercellular and cross-kingdom communications, suggesting that these nanostructures may contribute to distinct aspects of plant physiology such as development, defense, reproduction, symbiotic relationships, etc. These findings are challenging the traditional view of signaling in plants. EVs are probably involved in the phloem's transport system, since this vascular tissue plays a crucial role in translocating nutrients, defensive compounds, and informational signals throughout the plant. The collection of phloem is experimentally challenging because sap is under high turgor pressure inside the sieve elements, which have a small diameter and are hidden within the plant organs. The goals of this work are to develop new protocols that allow us to detect EVs for the first time in the phloem of the plants, and to isolate these nanovesicles for in-depth analysis and characterization. Our protocols describe two distinct methods to collect the phloem sap from rice and melon. The first method (Basic Protocol 1) involves ‘Aphid stylectomy by radiofrequency microcautery’ using rice plants and the aphid Sitobion avenae. This is considered the least invasive method for collecting phloem sap. The second method, ‘Stem incision’, involves cutting the stem of melon plants for collecting the exuded sap. Phloem sap EVs are then isolated by size exclusion chromatography. The results obtained in this study represent the first report on typical EVs isolated from in vivo–collected phloem sap.

The application of an OSMAC (One Strain–Many Compounds) approach on the fungus Pleotrichocladium opacum, isolated from a soil sample collected on the coast of Asturias (Spain), using different culture media, chemical elicitors, and cocultivation techniques resulted in the isolation and identification of nine new compounds (8, 9, 12, 15–18, 20, 21), along with 15 known ones (1–7, 10, 11, 14, 19, 22–25). Compounds 1–9 were detected in fungal extracts from JSA liquid fermentation, compounds 10–12 were isolated from a solid rice medium, whereas compounds 14 and 15 were isolated from a solid wheat medium. Addition of 5-azacytidine to the solid rice medium caused the accumulation of compounds 16–18, whereas adding N-acetyl-d-glucosamine triggered the production of two additional metabolites, 19 and 20. Finally, cocultivation of the fungus Pleotrichocladium opacum with Echinocatena sp. in a solid PDA medium led to the production of five additional natural products, 21–25. The structures of the new compounds were elucidated by HRESIMS and 1D and 2D NMR as well as by comparison with literature data. DP4+ and mix-J-DP4 computational methods were applied to determine the relative configurations of the novel compounds, and in some cases, the absolute configurations were assigned by a comparison of the optical rotations with those of related natural products.

Background: This study is based on a previous literature review of hydrotherapy techniques for patients with cerebral palsy (CP). This review found no significant differences between the different methods and techniques considered, and therefore a hydrotherapy protocol was developed, according to the model of psychomotor development achieved by a child in good health, in order to assess its efficacy in the treatment of children with CP. The hydrotherapy was oriented towards structural, functional and social aspects of the patient’s condition, within the overall rehabilitation process. Goals: To evaluate the changes observed in the gross motor function of children affected by CP, at baseline and after six months of hydrotherapy. Method: A prospective longitudinal study was conducted of 12 patients aged 18-33 months at the beginning (9 boys and 3 girls). The intervention protocol consisted of a 30-minute session performed once weekly, with 20 minutes in the pool. Gross motor function was assessed using the validated gross motor function measure (GMFM) scale prior to hydrotherapy and at six months after initiating the treatment. There was no control group, since the diverse symptoms typical of this pathology made it impractical. Instead, each patient constituted their own control, pre and post-test. Results: A statistically significant (p≤0.001) improvement was measured, on the GMFM global scale and on the following subscales: a) lying and rolling; b) sitting; c) crawling and kneeling; d) standing; e) walking, running and jumping. Conclusions: Hydrotherapy promotes the development of gross motor skills in children affected by CP. It also contributes to a positive development in the normal GMFM curve and a post-intervention improvement in the corresponding percentiles., Introduçao: O presente estúdio, parte de uma revisão bibliográfica anterior sobre as técnicas de tratamento em piscina para pacientes afetados pelo paralisia cerebral infantil (PC). Se você não encontrar diferenças diferenciadas entre os métodos e técnicas utilizadas, desenhe um programa de exercícios segundo o modelo de desenvolvimento psicomotor do filho sano, implemente e avalie a eficácia no tratamento de pessoas com esta patologia que usa o uso do hidroterapeuta orientado aspectos estruturais, funcionais e sociais no processo global de reabilitação. Objetivos: O objetivo deste estúdio é avaliar a evolução que produz a função motora grega nos menores afetados pelo PC, menos seis meses após o tratamento. Método: trata-se de um estúdio longitudinal prospectivo de 12 pacientes com idades compreendidas entre 18 e 33 meses de idade, 9 anos e 3 anos. O protocolo de tratamento consiste em uma sessão semanal de 30 minutos dos códigos, 20 a ser realizada diretamente na piscina. Se você avaliar usando a Escala de Função Motora Bruta (GMFM) validada antes de realizar a hidroterapia e os seis meses após o início da intervenção. Não há um grupo de controle usado que debita a sintomatologia apropriada dessa patologia para a desaconselhidão. Cada paciente é o controle adequado entre o pré e o pósteste. Resultados: Se você observar uma diferença estatisticamente significativa, na escala global GMFM e nos aspectos das subescalas: a) Tumbado e rodando, b) Sentado, c) Gatilho e rodillas, d) torta, e) Andando, corrigindo , saltando. Conclusões: A hidroterapia promove o desenvolvimento de habilidades motoras em animais afetados por PC. Também contribuímos com um projeto positivo na curva GMFM normal e em uma parte posterior da intervenção nos percentis correspondentes, Antecedentes: El presente estudio, parte de una revisión bibliográfica anterior sobre las técnicas de tratamiento en piscina para los pacientes afectados de Parálisis Cerebral Infantil (PC). Al no encontrar diferencias significativas entre los distintos métodos y técnicas utilizados, diseñamos un programa de ejercicios según el modelo del desarrollo psicomotor del niño sano, implementándolo y valorando su eficacia en el tratamiento de personas con esta patología haciendo un uso de la hidroterapia orientado hacia los aspectos estructurales, funcionales y sociales en el proceso global de la rehabilitación. Objetivos: El objetivo de este estudio es valorar la evolución que se produce en la función motora gruesa en niños afectados de PC, después de seis meses de tratamiento. Método: Se trata de un estudio longitudinal prospectivo sobre 12 pacientes con edades comprendidas entre 18 y 33 meses al comienzo, 9 niños y 3 niñas. El protocolo de tratamiento consiste en una sesión semanal de 30 minutos de los cuales, 20 se realizan íntegramente en piscina. Se evaluó mediante la Escala validada Gross Motor Function Measure (GMFM) antes de realizar hidroterapia y a los seis meses de iniciar la intervención. No se ha utilizado grupo control debido a que la diversa sintomatología propia de esta patología lo desaconseja. Cada paciente es su propio control entre el pre y post-test. Resultados: Se observa una mejoría, estadísticamente significativa, en la escala global GMFM y en aspectos específicos de las subescalas: a) Tumbado y rodando, b) Sentado, c) Gateando y de rodillas, d) de pie, e) Andando, corriendo, saltando. Conclusiones: La hidroterapia promueve el desarrollo de habilidades motoras gruesas en niños afectados por PC. También contribuye a un desarrollo positivo en la curva GMFM normal y una mejora posterior a la intervención en los percentiles correspondientes