RESUMO
The report of critical values ââis considered a necessary practice in clinical pathology laboratories, since its immediate communication determines therapeutic decisions that can save lives. The choice of critical values ââand the way in which they are reported must be a joint decision between the laboratory specialists and the clinical professionals of each health facility, since they must be integrated into the dynamics of the services involved. Among the critical values ââthat have been chosen for this review, we have included quantitative parameters, whose processing is carried out in automated hematology analyzers, and morphological findings in peripheral blood smears, which are evaluated by trained professionals.
El reporte de valores críticos se considera una práctica necesaria en los laboratorios de patología clínica, pues su comunicación inmediata determina decisiones terapéuticas que pueden salvar vidas. La elección de los valores críticos y el modo en que son reportados debe ser una decisión conjunta entre los especialistas del laboratorio y los profesionales clínicos de cada establecimiento de salud, ya que deben integrarse a la dinámica de los servicios involucrados. Entre los valores críticos que se han escogido para esta revisión hemos incluido parámetros cuantitativos, cuyo procesamiento se ejecuta en analizadores hematológicos automatizados, y hallazgos morfológicos en los frotis de sangre periférica, lo cuales son evaluados por profesionales capacitados.
RESUMO
Resilience of growing in arid and semiarid regions and a high capacity of accumulating sugar-rich biomass with low lignin percentages have placed Agave species as an emerging bioenergy crop. Although transcriptome sequencing of fiber-producing agave species has been explored, molecular bases that control wall cell biogenesis and metabolism in agave species are still poorly understood. Here, through RNAseq data mining, we reconstructed the cellulose biosynthesis pathway and the phenylpropanoid route producing lignin monomers in A. tequilana, and evaluated their expression patterns in silico and experimentally. Most of the orthologs retrieved showed differential expression levels when they were analyzed in different tissues with contrasting cellulose and lignin accumulation. Phylogenetic and structural motif analyses of putative CESA and CAD proteins allowed to identify those potentially involved with secondary cell wall formation. RT-qPCR assays revealed enhanced expression levels of AtqCAD5 and AtqCESA7 in parenchyma cells associated with extraxylary fibers, suggesting a mechanism of formation of sclerenchyma fibers in Agave similar to that reported for xylem cells in model eudicots. Overall, our results provide a framework for understanding molecular bases underlying cell wall biogenesis in Agave species studying mechanisms involving in leaf fiber development in monocots.
RESUMO
Together with their undeniable role in the ecology of arid and semiarid ecosystems, Agave species are emerging as a model to dissect the relationships between crassulacean acid metabolism and high efficiency of light and water use, and as an energy crop for bioethanol production. Transcriptome resources from economically valuable Agaves species, such as Agave tequilana and A. salmiana, as well as hybrids for fibers, are now available, and multiple gene expression landscape analyses have been reported. Key components in molecular mechanisms underlying drought tolerance could be uncovered by analyzing gene expression patterns of roots. This study describes an efficient protocol for high-quality total RNA isolation from phenolic compounds-rich Agave roots. Our methodology involves suitable root handling and collecting in the field and using saving-time commercial kits available. RNA isolated from roots free of lignified out-layers and clean cortex showed high values of quality and integrity according to electrophoresis and microfluidics-based platform. Synthesis of long full-length cDNAs and PCR amplification tested the suitability for downstream applications of extracted RNA. The protocol was applied successfully to A. tequilana roots but can be used for other Agave species that also develop lignified epidermis/exodermis in roots.
RESUMO
Spines are key plant modifications developed to deal against herbivores; however, its physical structure and chemical composition have been little explored in plant species. Here, we took advantage of high-throughput chromatography to characterize chemical composition of Agave fourcroydes Lem. spines, a species traditionally used for fiber extraction. Analyses of structural carbohydrate showed that spines have lower cellulose content than leaf fibers (52 and 72%, respectively) but contain more than 2-fold the hemicellulose and 1.5-fold pectin. Xylose and galacturonic acid were enriched in spines compared to fibers. The total lignin content in spines was 1.5-fold higher than those found in fibers, with elevated levels of syringyl (S) and guaiacyl (G) subunits but similar S/G ratios within tissues. Metabolomic profiling based on accurate mass spectrometry revealed the presence of phenolic compounds including quercetin, kaempferol, (+)-catechin, and (-)-epicatechin in A. fourcroydes spines, which were also detected in situ in spines tissues and could be implicated in the color of these plants' structures. Abundance of (+)-catechins could also explain proanthocyanidins found in spines. Agave spines may become a plant model to obtain more insights about cellulose and lignin interactions and condensed tannin deposition, which is valuable knowledge for the bioenergy industry and development of naturally dyed fibers, respectively.
RESUMO
RESUMEN En los últimos años, las células troncales mesenquimales (Mesenchymal Stem Cells, MSC) han adquirido mucha importancia debido a su gran plasticidad y su capacidad de liberar factores paracrinos con capacidad de interactuar con diversos tipos celulares, tejidos y órganos. El uso de MSC en medicina regenerativa es importante debido a que, al no expresar las moléculas del complejo mayor de histocompatibilidad (MHC) clase II ni moléculas co-estimuladoras y tener baja expresión del MHC clase I, haría que no sean rechazadas por individuos de la misma especie, es posible utilizarlas no sólo de manera autóloga, sino también, eventualmente, alogénica. Sin embargo, es importante demostrar científicamente muchas de sus propiedades, entre ellas las inmunomoduladoras. Al tener varias fuentes de obtención, se debe estandarizar la que sea la mejor para garantizar la pureza y calidad de las MSC. Finalmente, es importante que cuando se trabaje con estas células se demuestre completamente las características del cultivo celular, la inmunotipificación y su capacidad de diferenciación. Se están ensayando muchas aplicaciones clínicas de las MSC. Dentro de ellas, su capacidad para mejorar la recuperación y potencial curación de úlceras crónicas como las diabéticas, ha atraído la atención por su potencial impacto terapéutico.
ABSTRACT In recent years, mesenchymal stem cells (MSC) have become very important due to their high plasticity and their ability to release paracrine factors able to interact with various cell types, tissues and organs. The use of MSC in regenerative medicine became of vital importance, since they do not express histocompatibility MHC molecules class II nor costimulant molecules, and low expression of MHC class I, will not be rejected by individuals of same species, they could be used in an autologous, and eventually, allogeneic manner. However, it is important to scientifically demonstrate many properties, including immunomodulatory ones. Having several sources of obtaining, it should be standardized the best one to ensure the purity and quality of these cells. Finally, it is important when working with these cells, that characteristics of cell culture, immunophenotyping and differentiation capacity are fully demonstrated. MSC have been applied in several clinical uses. Among them, their ability to improve, and even heal chronic ulcers, as diabetic, has attracted attention for its potential therapeutic impact.
Assuntos
Humanos , Células-Tronco Mesenquimais , Transplante de Células-Tronco MesenquimaisRESUMO
OBJECTIVES.: To evaluate the healing effect of a Piper aduncum ethanol-water extract on an adult human dermal fibroblast cell line (hDFa). MATERIALS AND METHODS.: After obtaining the extract via solid-liquid extraction, concentration, and lyophilization, extract proteins were purified using reverse phase high-performance liquid chromatography, identified using tandem mass spectrometry of tryptic peptides, and analyzed using MALDI-TOF-TOF on an ABSciex4800 mass spectrometer. Half maximum effective concentration values (EC50), half maximum inhibiting concentration (IC50), and percentages of cell proliferation were determined using tetrazolium salt assays. Cell migration was evaluated using a "scratch assay". Growth factor expression in cells was analyzed via quantitative real-time reverse transcription polymerase chain reaction. RESULTS.: Against the hDFa cell line, the extract had an IC50 of 200 µg/mL and EC50 of 103.5 µg/mL. In the proliferation assay, protein K2 (obtained from the extract) exhibited increased proliferative activity relative to other treatments (1 µg/mL); this agent also exhibited increased activity (50 µg/mL) in the fibroblast migration assay.Furthermore, the relative expression of platelet-derived growth factor increased by 8.6-fold in the presence of K2 protein relative to the control. CONCLUSIONS.: The hydroethanolic extract of Piper aduncum and its component proteins increased the proliferation and migration of hDFa and increased the expression of growth factors involved in the healing process.
Assuntos
Fibroblastos/efeitos dos fármacos , Piper/química , Extratos Vegetais/farmacologia , Proliferação de Células , Etanol , HumanosRESUMO
RESUMEN Objetivos. Evaluar el efecto cicatrizante del extracto hidroetanólico de Piper aduncum, en una línea celular de fibroblastos Dermales Adultos Humanos (hDFa). Materiales y métodos. El extracto se obtuvo mediante extracción sólido-líquido, fue concentrado y liofilizado. Se purificaron las proteínas del extracto mediante cromatografía líquida de alta eficacia de fase reversa (RP-HPLC); las proteínas fueron identificadas por espectrometría de masas en tándem de péptidos trípticos y se analizaron por MALDI-TOF-TOF en un espectrómetro de masa ABSciex4800. Los valores de concentración efectiva media (EC50), concentración inhibitoria media (IC50), y el porcentaje de proliferación celular; fueron determinados por ensayos con sales de tetrazolio (MTT) . La migración celular se evaluó mediante la "técnica de rayado" . Se analizó la expresión de factores de crecimiento mediante el ensayo de reacción en cadena de la polimerasa con transcriptasa reversa a tiempo real (RT- qPCR). Resultados. La línea hDFa evidenció un IC50 de 200 µg/mL con el extracto, el valor de EC50 fue 103,5 µg/mL. En el ensayo de proliferación, la proteína K2; mostró mayor actividad en la proliferación respecto de otros tratamientos (1 µg/mL). En el ensayo de migración de fibroblastos, la proteína K2 mostró mayor actividad (50 µg/mL). La expresión relativa del factor de crecimiento derivado de plaquetas (PDGF) se incrementó 8,6 veces respecto al control, en presencia de la proteína K2. Conclusiones. El extracto hidroetanólico, de Piper aduncum, así como las proteínas que contiene, incrementaron la proliferación y migración de fibroblastos dermales humanos (hDFa); así mismo, aumentaron la expresión de factores de crecimiento que intervienen en el proceso de cicatrización.
ABSTRACT Objectives. To evaluate the healing effect of a Piper aduncum ethanol-water extract on an adult human dermal fibroblast cell line (hDFa). Materials and Methods. After obtaining the extract via solid-liquid extraction, concentration, and lyophilization, extract proteins were purified using reverse phase high-performance liquid chromatography, identified using tandem mass spectrometry of tryptic peptides, and analyzed using MALDI-TOF-TOF on an ABSciex4800 mass spectrometer. Half maximum effective concentration values (EC50), half maximum inhibiting concentration (IC50), and percentages of cell proliferation were determined using tetrazolium salt assays. Cell migration was evaluated using a "scratch assay". Growth factor expression in cells was analyzed via quantitative real-time reverse transcription polymerase chain reaction. Results. Against the hDFa cell line, the extract had an IC50 of 200 μg/mL and EC50 of 103.5 µg/mL. In the proliferation assay, protein K2 (obtained from the extract) exhibited increased proliferative activity relative to other treatments (1 µg/mL); this agent also exhibited increased activity (50 µg/mL) in the fibroblast migration assay.Furthermore, the relative expression of platelet-derived growth factor increased by 8.6-fold in the presence of K2 protein relative to the control. Conclusions. The hydroethanolic extract of Piper aduncum and its component proteins increased the proliferation and migration of hDFa and increased the expression of growth factors involved in the healing process.
Assuntos
Humanos , Extratos Vegetais/farmacologia , Piper/química , Fibroblastos/efeitos dos fármacos , Proliferação de Células , EtanolRESUMO
In recent years, mesenchymal stem cells (MSC) have become very important due to their high plasticity and their ability to release paracrine factors able to interact with various cell types, tissues and organs. The use of MSC in regenerative medicine became of vital importance, since they do not express histocompatibility MHC molecules class II nor costimulant molecules, and low expression of MHC class I, will not be rejected by individuals of same species, they could be used in an autologous, and eventually, allogeneic manner. However, it is important to scientifically demonstrate many properties, including immunomodulatory ones. Having several sources of obtaining, it should be standardized the best one to ensure the purity and quality of these cells. Finally, it is important when working with these cells, that characteristics of cell culture, immunophenotyping and differentiation capacity are fully demonstrated. MSC have been applied in several clinical uses. Among them, their ability to improve, and even heal chronic ulcers, as diabetic, has attracted attention for its potential therapeutic impact.
Assuntos
Células-Tronco Mesenquimais , Humanos , Transplante de Células-Tronco MesenquimaisRESUMO
We assessed in vivo the anti-inflammatory activity of two Cat's claw bark extracts, by comparing a spray-dried hydroalcoholic extract against an aqueous freeze-dried extract, to determine which extract was more effective. We used the carrageenan-induced paw edema model in mice. In addition, to assess the molecular mechanism of action, we determined the inhibition of NF-kappa B through the Electrophoretic Mobility Shift Assay (EMSA) and the effects on cycloxygenase-1 and -2. Results showed that the anti-inflammatory activity was significantly higher using the hydroalcoholic compared with the aqueous extract (P<0.05). The extracts also showed little inhibitory activity on cyclooxygenase-1 and -2. It cannot be excluded that the slight inhibitory activity on DNA binding of NF-kappa B is due to cytotoxic effects.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Unha-de-Gato , Animais , Anti-Inflamatórios não Esteroides/isolamento & purificação , Ciclo-Oxigenase 1 , Ciclo-Oxigenase 2 , Edema/induzido quimicamente , Edema/tratamento farmacológico , Inibidores Enzimáticos/farmacologia , Feminino , Humanos , Isoenzimas/antagonistas & inibidores , Isoenzimas/metabolismo , Células Jurkat/efeitos dos fármacos , Proteínas de Membrana , Camundongos , Camundongos Endogâmicos BALB C , NF-kappa B/metabolismo , Casca de Planta , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Prostaglandina-Endoperóxido Sintases/metabolismoRESUMO
La Maca (lepidium meyeni) es una planta peruana conocida desde la época precolombina por una serie de efectos nutricionales y medicinales. Uno de estos efectos reportados tradicionales son sus propiedades para reducir la tensión, el estrés y la depresión. Científicamente se han demostrado sus cualidades nutricionales, su efecto favorecedor de la fertilidad y sus propiedades energizantes. Igualmente, en su composición química se ha demostrado la presencia de varios compuestos capaces de tener efectos biológicos en los seres vivos, como glucosinolatos, esteroles, saponinas, entre otros. Su efecto mejorador del estrés y la tensión, sin embargo, no ha sido aún estudiada científicamente, lo cual fue motivo de este estudio. Se usaron ratones raza Swiss, los cuales han sido alimentados desde el momento de su ablactancia con una dieta diferenciada divida en tres grupos: A) Grupo control alimentado con una dieta balanceada comercial habitual; b) Grupo maca cruda: alimentados con una mezcla de la dieta balanceada comercial habitual igual que el grupo A, pero a la cual se le había reemplazado un 30 por ciento por Maca Cruda; y, c) Grupo Maca Cocida: alimentados con la dieta balanceada a la cual se le reemplazo un 30 por ciento con Maca Cocida. Luego de 15 semanas de alimentación con esta dieta diferenciada, los animales fueron individualmente sometidos a 6 sesiones de estimulación de estrés con descargas eléctricas no letales y se cuantificaron indicadores de neuropticismo como: características de los ojos, el pelaje, manifestaciones gastrointestinales y vesicales, vasotensión periférica, movilidad del animal, excitación emocional y hábitos alimenticios. Los resultados mostraron que los animales suplementados con Maca (sea esta cruda o cocida) tuvieron una mayor resistencia al desarrollo de signos de neuroticismo, lograron menores puntajes de neuroticismo y tuvieron desaparición de los signos de neuroticismo más rápidamente que los animales que no recibieron Maca, con una diferencia que en todos los casos fue estadísticamente significativa (Análisis de varianza, Kruskal-Wallis, p menor 0,05). A nuestro conocimiento este es el primer estudio que demuestra científicamente las propiedades anti-estrés de la Maca. Se requiere mayores estudios para ratificar esta observación.