Development and carotenoid synthesis in dark-grown carrot taproots require PHYTOCHROME RAPIDLY REGULATED1.

Light stimulates carotenoid synthesis in plants during photomorphogenesis through the expression of PHYTOENE SYNTHASE (PSY), a key gene in carotenoid biosynthesis. The orange carrot (Daucus carota) synthesizes and accumulates high amounts of carotenoids in the taproot that grows underground. Contrary to other organs, light impairs carrot taproot development and represses the expression of carotenogenic genes, such as DcPSY1 and DcPSY2, reducing carotenoid accumulation. By means of RNA sequencing, in a previous analysis, we observed that carrot PHYTOCHROME RAPIDLY REGULATED1 (DcPAR1) is more highly expressed in the underground grown taproot compared with those grown in light. PAR1 is a transcriptional cofactor with a negative role in shade avoidance syndrome regulation in Arabidopsis (Arabidopsis thaliana) through the dimerization with PHYTOCHROME-INTERACTING FACTORs (PIFs), allowing a moderate synthesis of carotenoids. Here, we show that overexpressing AtPAR1 in carrot increases carotenoid production in taproots grown underground as well as DcPSY1 expression. The high expression of AtPAR1 and DcPAR1 led us to hypothesize a functional role of DcPAR1 that was verified through in vivo binding to AtPIF7 and overexpression in Arabidopsis, where AtPSY expression and carotenoid accumulation increased together with a photomorphogenic phenotype. Finally, DcPAR1 antisense carrot lines presented a dramatic decrease in carotenoid levels and in relative expression of key carotenogenic genes as well as impaired taproot development. These results suggest that DcPAR1 is a key factor for secondary root development and carotenoid synthesis in carrot taproot grown underground.

Carrot DcALFIN4 and DcALFIN7 Transcription Factors Boost Carotenoid Levels and Participate Differentially in Salt Stress Tolerance When Expressed in Arabidopsis thaliana and Actinidia deliciosa.

ALFIN-like transcription factors (ALs) are involved in several physiological processes such as seed germination, root development and abiotic stress responses in plants. In carrot (Daucus carota), the expression of DcPSY2, a gene encoding phytoene synthase required for carotenoid biosynthesis, is induced after salt and abscisic acid (ABA) treatment. Interestingly, the DcPSY2 promoter contains multiple ALFIN response elements. By in silico analysis, we identified two putative genes with the molecular characteristics of ALs, DcAL4 and DcAL7, in the carrot transcriptome. These genes encode nuclear proteins that transactivate reporter genes and bind to the carrot DcPSY2 promoter in yeast. The expression of both genes is induced in carrot under salt stress, especially DcAL4 which also responds to ABA treatment. Transgenic homozygous T3 Arabidopsis thaliana lines that stably express DcAL4 and DcAL7 show a higher survival rate with respect to control plants after chronic salt stress. Of note is that DcAL4 lines present a better performance in salt treatments, correlating with the expression level of DcAL4, AtPSY and AtDXR and an increase in carotenoid and chlorophyll contents. Likewise, DcAL4 transgenic kiwi (Actinidia deliciosa) lines show increased carotenoid and chlorophyll content and higher survival rate compared to control plants after chronic salt treatment. Therefore, DcAL4 and DcAL7 encode functional transcription factors, while ectopic expression of DcAL4 provides increased tolerance to salinity in Arabidopsis and Kiwi plants.

A de novo transcriptome analysis revealed that photomorphogenic genes are required for carotenoid synthesis in the dark-grown carrot taproot.

Carotenoids are terpenoid pigments synthesized by all photosynthetic and some non-photosynthetic organisms. In plants, these lipophilic compounds are involved in photosynthesis, photoprotection, and phytohormone synthesis. In plants, carotenoid biosynthesis is induced by several environmental factors such as light including photoreceptors, such as phytochromes (PHYs) and negatively regulated by phytochrome interacting factors (PIFs). Daucus carota (carrot) is one of the few plant species that synthesize and accumulate carotenoids in the storage root that grows in darkness. Contrary to other plants, light inhibits secondary root growth and carotenoid accumulation suggesting the existence of new mechanisms repressed by light that regulate both processes. To identify genes induced by dark and repressed by light that regulate carotenoid synthesis and carrot root development, in this work an RNA-Seq analysis was performed from dark- and light-grown carrot roots. Using this high-throughput sequencing methodology, a de novo transcriptome model with 63,164 contigs was obtained, from which 18,488 were differentially expressed (DEG) between the two experimental conditions. Interestingly, light-regulated genes are preferably expressed in dark-grown roots. Enrichment analysis of GO terms with DEGs genes, validation of the transcriptome model and DEG analysis through qPCR allow us to hypothesize that genes involved in photomorphogenesis and light perception such as PHYA, PHYB, PIF3, PAR1, CRY2, FYH3, FAR1 and COP1 participate in the synthesis of carotenoids and carrot storage root development.

Organizing pneumonia as presenting feature of primary Sjögren's syndrome: A case report.

Sjögren's Syndrome (SS) is an autoimmune inflammatory disease, characterized by lymphocytic infiltration of exocrine glands. Approximately 10% of patients with SS have pulmonary involvement as the first manifestation of their disease, the most common being non-specific interstitial pneumonia. We present the case of a 51-year-old man with organizing pneumonia as the presenting feature of primary SS. Pulmonary involvement as the presenting feature of SS is uncommon, especially when the pattern on CT-scan is that of organized pneumonia. Initial management includes steroids and other immunosuppressants agents, with a better response in organized pneumonia secondary SS cases.

Putative Daucus carota Capsanthin-Capsorubin Synthase (DcCCS) Possesses Lycopene ß-Cyclase Activity, Boosts Carotenoid Levels, and Increases Salt Tolerance in Heterologous Plants.

Plant carotenoids are synthesized and accumulated in plastids through a highly regulated pathway. Lycopene ß-cyclase (LCYB) is a key enzyme involved directly in the synthesis of α-carotene and ß-carotene through the cyclization of trans-lycopene. Daucus carota harbors two LCYB genes, of which DcLCYB2 (annotated as CCS-Like) is mostly expressed in mature storage roots, an organ that accumulates high α-carotene and ß-carotene content. In this work, we determined that DcLCYB2 of the orange Nantes variety presents plastid localization and encodes for a functional LCYB enzyme determined by means of heterologous complementation in Escherichia coli. Also, ectopic expression of DcLCYB2 in tobacco (Nicotiana tabacum) and kiwi (Actinidia deliciosa) plants increases total carotenoid content showing its functional role in plants. In addition, transgenic tobacco T2 homozygous plants showed better performance under chronic salt treatment, while kiwi transgenic calli also presented a higher survival rate under salt treatments than control calli. Our results allow us to propose DcLCYB2 as a prime candidate to engineer carotenoid biofortified crops as well as crops resilient to saline environments.

Daucus carota DcPSY2 and DcLCYB1 as Tools for Carotenoid Metabolic Engineering to Improve the Nutritional Value of Fruits.

Carotenoids are pigments with important nutritional value in the human diet. As antioxidant molecules, they act as scavengers of free radicals enhancing immunity and preventing cancer and cardiovascular diseases. Moreover, α-carotene and ß-carotene, the main carotenoids of carrots (Daucus carota) are precursors of vitamin A, whose deficiency in the diet can trigger night blindness and macular degeneration. With the aim of increasing the carotenoid content in fruit flesh, three key genes of the carotenoid pathway, phytoene synthase (DcPSY2) and lycopene cyclase (DcLCYB1) from carrots, and carotene desaturase (XdCrtI) from the yeast Xanthophyllomyces dendrorhous, were optimized for expression in apple and cloned under the Solanum chilense (tomatillo) polygalacturonase (PG) fruit specific promoter. A biotechnological platform was generated and functionally tested by subcellular localization, and single, double and triple combinations were both stably transformed in tomatoes (Solanum lycopersicum var. Microtom) and transiently transformed in Fuji apple fruit flesh (Malus domestica). We demonstrated the functionality of the S. chilense PG promoter by directing the expression of the transgenes specifically to fruits. Transgenic tomato fruits expressing DcPSY2, DcLCYB1, and DcPSY2-XdCRTI, produced 1.34, 2.0, and 1.99-fold more total carotenoids than wild-type fruits, respectively. Furthermore, transgenic tomatoes expressing DcLCYB1, DcPSY2-XdCRTI, and DcPSY2-XdCRTI-DcLCYB1 exhibited an increment in ß-carotene levels of 2.5, 3.0, and 2.57-fold in comparison with wild-type fruits, respectively. Additionally, Fuji apple flesh agroinfiltrated with DcPSY2 and DcLCYB1 constructs showed a significant increase of 2.75 and 3.11-fold in total carotenoids and 5.11 and 5.84-fold in ß-carotene, respectively whereas the expression of DcPSY2-XdCRTI and DcPSY2-XdCRTI-DcLCYB1 generated lower, but significant changes in the carotenoid profile of infiltrated apple flesh. The results in apple demonstrate that DcPSY2 and DcLCYB1 are suitable biotechnological genes to increase the carotenoid content in fruits of species with reduced amounts of these pigments.

Non-random medicinal plants selection in the Kichwa community of the Ecuadorian Amazon.

The non-random selection of medicinal plants theory, which predicts taxonomical biases in ethnopharmacopeias, indirectly demonstrates that traditional medicinal systems are rational and based in part on the therapeutic efficacy of plants. This theory suggests that because members of a taxonomical group share similar characteristics, some groups will be over-utilized in pharmacopeias, while other groups bereft of therapeutic potential will be under-utilized medicinally. Empirical evidence fo this theory comes from studies that used data collected at the national level which may lead to the overestimation of medicinal plant list given that some parts of the country (e.g., protected areas) can be unavailable for medicinal plant collection. Similarly, because medicinal plant importance and knowledge can be gender-specific and depends on the degree of exposure of a community, failure to account for gender and community experience can limit our understanding of non-random selection of medicinal plants. In this study, we used the negative binomial model and an examination of studentized residuals to demonstrate that a Kichwa community in the Ecuadorian Amazon over-utilized different sets of medicinal plant families depending on the gender of the informants or the experience of the community. We showed that utilizing local data instead of nationwide data reveals new over-utilized families. Seven of the nine most over-utilized medicinal plant families we found were previously reported in different biogeographical regions. The other two families are novel reports. Overall, our study proposes a novel method to uncover the intracultural heterogeneity of traditional knowledge and people non-random selection of medicinal plants at the local level.

Differential role of the two ζ-carotene desaturase paralogs in carrot (Daucus carota): ZDS1 is a functional gene essential for plant development and carotenoid synthesis.

Daucus carota is a biennale crop that develops an edible storage root. Orange carrots, the most consumed cultivar worldwide, accumulate high levels of ß-carotene and α-carotene in the storage root during secondary growth. Genes involved in ß-carotene synthesis have been identified in carrots and unlike most species, D. carota has two ζ-carotene desaturase genes, named ZDS1 and ZDS2, that share 91.3 % identity in their coding regions. ZDS1 expression falls during leaf, but not root development, while ZDS2 is induced in leaves and storage roots of a mature plant. In this work, by means of post-transcriptional gene silencing, we determined that ZDS1 is essential for initial carrot development. The suppression of the expression of this gene by RNAi triggered a reduction in the transcript levels of ZDS2 and PSY2 genes, with a concomitant decrease in the carotenoid content in both, leaves and storage roots. On the contrary, transgenic lines with reduced ZDS2 transcript abundance maintain the same levels of expression of endogenous ZDS1 and PSY2 and carotenoid profile as wild-type plants. The simultaneous silencing of ZDS1 and ZDS2 resulted in lines with a negligible leaf and root development, as well as significantly lower endogenous PSY2 expression. Further functional analyses, such as a plastidial subcellular localization of ZDS1:GFP and the increment in carotenoid content in transgenic tobacco plants overexpressing the carrot ZDS1, confirmed that ZDS1 codifies for a functional enzyme. Overall, these results lead us to propose that the main ζ-carotene desaturase activity in carrot is encoded by the ZDS1 gene and ZDS2 gene has a complementary and non essential role.

Endobronchial ultrasound-guided transbronchial needle aspirate for diagnosis of anaplastic large cell lymphoma of unusual presentation: A case report.

Anaplastic large cell lymphoma (ALCL) is a rare type of non-Hodgkin lymphoma (NHL) originated from mature post thymic T cells. They represent 1-3% of NHL. Different subtypes have been described: Anaplastic lymphoma kinase (ALK)-negative ALCL, ALK-positive ALCL and breast implant-associated ALCL. ALK-positive ALCL affects mainly the young and has better prognosis. We present a case report of an adult woman with AKL-positive ALCL, diagnosed by endobronchial ultrasound-guided transbronchial needle aspirate (EBUS-TBNA). A 59-year-old women with no history of breast implants, was admitted for a four-month low back pain. Initially, the patient was treated for a spondyloarthropathy, but due to persistence of the symptoms, a lumbosacral MRI was performed, showing changes in morphology and signal intensity in the vertebral body of L3, along with edema and a paravertebral collection that affected the left psoas muscle, suggesting granulomatous spondylodiscitis. Chest CT-scan showed mild left pleural effusion, subcarinal and right hiliar adenomegalies. An EBUS-TBNA with ROSE (rapid on-site evaluation) was performed showing positive findings for malignancy, suggestive of hematolymphoid neoplasia. Pathology analysis showed an AKL-positive ALCL. Additionally, a biopsy of paravertebral tissue biopsy was obtained, which was consistent with the nodal sample. Chemotherapy was initiated with the CHOP protocol: cyclophosphamide, hydroxydaunorubicin, vincristine sulfate and prednisone. EBUS-TBNA is a minimally invasive and safe technique for obtaining mediastinal samples. Collaboration with a cytopathologist trained to perform ROSE improves the diagnostic performance.

Procesos institucionales para la atención del paciente sospechoso o diagnosticado con Covid-19 / Institutional processes for the care of suspected or diagnosed COVID-19 patients / Processos institucionais para atendimento de pacientes suspeitos ou diagnosticados com Covid-19

Objetivo: Identificar los procesos institucionales implementados y/o modificados para la at-ención de pacientes sospechosos o diagnosticados con COVID-19 en una institución de salud de Cundinamarca. Materiales y métodos: Se realizó un estudio trasversal descriptivo. La recolección de datos se efectuó a través del método de revisión de registros de los procesos institucionales de una institución de segundo nivel de complejidad, desde marzo del 2020 hasta diciembre del 2021. A continuación, la información se sistematizó en una matriz de Excel previo diseño, validación y aplicación. Resultados: Se precisa que como dato princi-pal la institución cuenta con 80 procesos establecidos de los cuales 41 fueron modificados, lo que corresponde al 51.25%; 24 no sufrieron ninguna modificación por la pandemia lo que equivale al 30%; también se implementaron15 procesos nuevos que corresponden a un 18.75%. Conclusiones: Se pudo determinar que las principales modificaciones se observaron en la presencia de un mayor número de procesos modificados, puesto que ante la necesidad inmediata se requirió dar respuesta a la contingencia mediante la adaptación de lineamientos ya estandarizados anteriormente, lo que permitió agilizar la respuesta ante el suceso. Además, se evidenció un impacto significativo en la atención ambulatoria debido a la demanda del ser-vicio, así como en los procesos dirigidos directamente a la atención del paciente. Se observó que el impacto del COVID-19 transformó significativamente los procesos implementados por las entidades sanitarias, lo que las llevó a realizar cambios en las acciones ejecutadas por el personal de la salud; de ahí, que este suceso permitió examinar las problemáticas impor-tantes que se habían invisibilizado

Objective: Identify the implemented and/or modified institutional processes for the attention of suspected or diagnosed COVID-19 patients in a health institution of Cundinamarca. Ma-terials and methods: A cross-sectional descriptive study was carried-out. The recollection of data was made through a review method of records of the institutional processes from an institution of second level of complexity, from March of 2020 to December of 2021. Next, the information was systematized in an Excel matrix that was previously designed, validated and applied. Results: It must be noted that as the main information, the institution has 80 established processes of which 41 were modified, corresponding to 51,25%; 24 did not have any modifications due to the pandemic, which is equivalent to 30%; also implementing 15 new processes that correspond to 18,75%. Conclusions: It was possible to determine that the main modifications were observed in the presence of a greater number of modified processes, since there was an immediate need that required to respond to the eventuality through the adaptation of previously standardized guidelines, which allowed to speed up the response to the event. Also, a significant impact was evidenced in ambulatory care due to the demand for the service, as well as in processes that were directly aimed at patient care. It was observed that the impact of COVID-19 significantly transformed the processes implemented by health entities, which led to make changes in the actions taken by the healthcare personnel; from there, this event allowed to examine important issues that had been made invisible

Objetivo: Identificar os processos institucionais implementados e/ou modificados para o at-endimento de pacientes suspeitos ou diagnosticados com a COVID-19 em uma instituição de saúde em Cundinamarca. Materiais e métodos: Foi realizado um estudo transversal descriti-vo. A coleta de dados foi realizada por meio do método de revisão dos registros dos processos institucionais de uma instituição de segundo nível de março de 2020 a dezembro de 2021. As informações foram então sistematizadas em Excel após validação e aplicação. Resulta-dos: a instituição possui 80 processos estabelecidos dos quais 41 foram modificados, o que corresponde a 51,25%; 24 (30%) não sofreram nenhuma modificação devido à pandemia; 15 (18,75%) novos processos também foram implementados. Conclusões: Foi possível de-terminar que houve maior número de processos modificados, uma vez que foi necessária a resposta à contingencia por meio da adaptação de diretrizes já padronizadas anteriormente, o que permitiu acelerar a resposta ao evento. Além disso, foi evidenciado um impacto significa-tivo no atendimento ambulatorial devido à demanda pelo serviço, bem como em processos voltados diretamente ao atendimento ao paciente. Observou-se que o impacto da COVID-19 transformou significativamente os processos implementados pelas entidades de saúde, o que os levou a fazer mudanças nas ações realizadas pelos profissionais de saúde; portanto, este evento tornou possível examinar os problemas importantes que tinham se tornado invisíveis

Mejoramiento de la calidad de la alimentación servida en una organización no gubernamental-red nacional de cuido, San José Costa Rica, 2016 / Improvement in the quality of food served in a non-governmental organization-national childcare network, San José, Costa Rica, 2016

RESUMEN Una buena alimentación durante la niñez es indispensable para un crecimiento y desarrollo óptimo. Por lo que se desarrolló una propuesta de intervención para mejorar la calidad de la alimentación servida a los niños y niñas de 2 a 12 años del Centro de Atención Integral Asociación Teen Challenge, que forma parte de la Red Nacional de Cuido de Costa Rica. El estudio se desarrolló en cinco etapas: diagnóstico, priorización de problemas, diseño de la propuesta de mejora, implementación y evaluación, durante 2016. Los datos se analizaron según los lineamientos nacionales e internacionales relacionados con la calidad: higiénica y sanitaria, bromatológica y sensorial. Se diseñó una propuesta de mejora de 10 sesiones educativas dirigida al personal de atención directa, con 14 estándares de calidad y se planteó la meta de cumplir 11 estándares. Al finalizar, la calidad de la alimentación mejoró; sin embargo, no se cumplió la meta. La realización de este tipo de proyectos es de suma importancia para la salud y bienestar de los menores de edad. La baja presencia de profesionales en Nutrición, poca capacitación y nula supervisión influyen negativamente en la calidad de la alimentación ofrecida en estas instituciones.

ABSTRACT Good nutrition during childhood is essential for optimal growth. For this reason, an intervention proposal was developed to improve the quality of food served to children ages 2 to 12 years at the Teen Challenge Child Care Center, which belongs to the National Child Care Network of Costa Rica. The study was developed in five stages: diagnosis, problem prioritization, intervention program design, implementation and evaluation during 2016. The data was analyzed according to national and international food quality guidelines such as: food safety, bromatological and sensorial properties. The main aspects that were improved were hygiene, food safety and bromatological food quality. A proposal of 10 educational sessions was designed, to be provided to the personnel of the center including teachers and kitchen staff, with 14 standards of quality and the goal was to comply with at least 11 of them. The quality of food served was improved; however, the target was not met. Studies of this type are of high importance for the health and well-being of young people. The low presence of nutrition professionals, lack of training and no supervision, negatively influence the quality of food offered in these spaces.

Consumo de alcohol en entornos urbanos alrededor de una universidad en Bogotá. Resultados de un estudio piloto / Consume of Alcohol in Urban Settings around a University in Bogotá. Results of a Pilot Study

Introducción: El objetivo de este estudio piloto fue caracterizar las percepciones de estudiantes universitarios sobre el consumo de alcohol y la disponibilidad de establecimientos que expenden licor en el perímetro urbano de una universidad de Bogotá. Métodos: Se llevó a cabo un estudio transversal y mixto en 201 estudiantes seleccionados por una muestra no probabilística. Se usó una encuesta autodiligenciada para evaluar percepciones respecto al consumo de alcohol alrededor de la universidad y sistemas de información geográfica para identificar los establecimientos que vendían alcohol. Resultados: El 66% de los participantes informó haber consumido alcohol alrededor del campus universitario. El 26% se mostraba de acuerdo o muy de acuerdo con la implementación de medidas estrictas para restringir el consumo, siendo mayor en personas que no consumieron (41,2%). El análisis geográfico encontró un patrón de concentración de oferta y consumo de alcohol en las inmediaciones de la universidad. Conclusión: Este estudio muestra la potencial relevancia que tiene la oferta y consumo de alcohol alrededor de un campus universitario para la salud pública. Se deben llevar a cabo estudios complementarios para entender, de mejor manera, posibles vínculos entre contextos urbanos universitarios y consumo de alcohol.

Background: The aim of this pilot study was to characterize the perceptions of university students in relation to alcohol consumption and the availability of alcohol outlets in the urban perimeter of a university in Bogotá. Methods: A mixed cross-sectional study was conducted among 201 undergraduate students ages 18 to 27 years using a non-probabilistic sampling design. The perceptions regarding alcohol consumption in the university's surroundings were assessed via a self-reported questionnaire. Geographic Information Systems were used to identify the outlets that sell alcohol. Results: 66% of participants reported alcohol consumption in in the university's surroundings. The proportion of students who agreed with implementing strict measures to control alcohol consumption was 26%, with higher percentages in those who didn't consume alcohol (41,2%). Geographic analysis showed that several urban attributes were correlated with alcohol consumption. Conclusión: The results of this study show the potential relevance that has the availability' of alcohol outlets near a university' campus as a public health issue. Further studies should be conducted in order to better understand the links between urban contexts around universities and alcohol consumption.

Una experiencia grupal con adolescentes con diagnóstico presuntivo de psicosis en consultorios externos de una institución pública: un punto de partida / A group experience with adolescents with presumptive diagnosis of psychosis in outpatient clinics of a public institution: a starting point

La sanción de la Ley Nacional de Salud Mental ha constituido un hito legislativo en el país en lo referente a la protección de los derechos de los usuarios de servicios de salud mental, adecuando a la Argentina a los tratados y principios internacionales sobre la materia. No obstante, son numerosos los debates y controversias en torno a su actual proceso de implementación y cumplimiento. El presente trabajo tiene como objetivo general describir y analizar la perspectiva de usuarios de servicios de salud mental acerca del proceso de implementación de la Ley y los desafíos para su cumplimiento. Se presentan parte de los resultados de un estudio más amplio enfocado en las perspecti-vas de usuarios participantes de una asociación centrada en la defensa de sus derechos. Se realizaron entrevistas semi estructuradas con ocho usuarios participantes de dicha asociación. Si bien destacan un avance a partir del derecho adquirido a participar a tra-vés de instancias concretas creadas por la ley, llaman la atención sobre las dificultades persistentes de acceso a derechos sociales (vivienda y trabajo) y, adicionalmente, de acceso al derecho a recibir la atención que menos restrinja sus derechos y libertades.