Antioxidant and Inhibitory Capacity of Tomato Leaf Ethanolic Extract against Streptococcus mutans, Porphyromonas gingivalis, and Candida albicans / Capacidad Antioxidante e Inhibitoria del Extracto Etanólico de la Hoja de Tomate contra Streptococcus mutans, Porphyromonas gingivalis y Candida albicans

Background: Tomato is a source of bioactive compounds, antimicrobials, and antioxidants. Tomato leaf preparations have been empirically used for anti-inflammatory, analgesic, antibiotic, and antiseptic purposes. However, research on the potential activity of tomato leaf extracts against oral microorganisms and in managing oropharyngeal infections is scarce. Objective: To investigate tomato leaf ethanolic extract's antioxidant and growth inhibitory capacity against common oral pathogenic microorganisms, namely, Streptococcus mutans, Porphyromonas gingivalis, and Candida albicans.Methods: Ethanolic extracts were made from 'Chonto' tomato (Lycopersicon esculentum) leaves. The antimicrobial activity was measured with the microdilution technique using vancomycin and fluconazole as positive controls. The antioxidant capacity was measured with the ORAC assay using Trolox as a positive control. Results: We found a high percentage of growth inhibition (≥100%) against S. mutans and P. gingivalis at a concentration of 500 mg/L. However, the extract was ineffective in inhibiting the growth of C. albicans. Finally, we observed that the extract exerted a high antioxidant capacity (126%) compared to the positive control. Conclusions: This study provides new insights into the potential antimicrobial effect of tomato leaf extracts on common oral pathogenic bacteria, which may ultimately result in the development of new herbal products that might help prevent and treat oral infections, such as dental caries and periodontal disease. Our findings also support previous studies on the high antioxidant capacity of tomato leaf extracts

Antecedentes: El tomate es una fuente de compuestos bioactivos, antimicrobianos y antioxidantes. Las hojas de tomate se han utilizado empíricamente con fines antiinflamatorios, analgésicos, antibióticos y antisépticos. Sin embargo, los estudios sobre la actividad de los extractos de hojas de tomate contra los microorganismos orales y en el manejo de las infecciones orofaríngeas son escasos. Objetivo: Investigar la capacidad antioxidante del extracto etanólico de la hoja de tomate y su actividad inhibitoria de crecimiento contra microorganismos patógenos orales comunes, a saber, Streptococcus mutans, Porphyromonas gingivalis y Candida albicans.Métodos: Se realizaron extractos etanólicos a partir de hojas de tomate 'Chonto' (Lycopersicon esculentum). La actividad antimicrobiana se midió con la técnica de microdilución utilizando vancomicina y fluconazol como controles positivos. La capacidad antioxidante se midió con el ensayo ORAC utilizando Trolox como control positivo. Resultados: Encontramos un alto porcentaje de inhibición del crecimiento (≥100%) contra a S. mutans y P. gingivalis a una concentración de 500 mg/L. Sin embargo, el extracto fue ineficaz en la inhibición el crecimiento de C. albicans. Finalmente, observamos que el extracto tuvo una alta capacidad antioxidante (126%) en comparación con el control positivo. Conclusiones: Este estudio proporciona nuevos conocimientos sobre el posible efecto antimicrobiano de los extractos de hojas de tomate en bacterias patógenas orales comunes, lo cual puede resultar en el desarrollo de nuevos productos naturales que podrían ayudar a prevenir y tratar infecciones orales, como la caries dental y la enfermedad periodontal. Nuestros hallazgos también respaldan los estudios previos sobre la alta capacidad antioxidante de los extractos de hojas de tomate

Nutritional quality, nutrient uptake and biomass production of Pennisetum purpureum cv. King grass.

The research was conducted to determine the effects of cutting interval and fertilization on the nutritional quality, nutrient uptake, and biomass production of King grass. The experimental design was a randomized complete block, using 4 blocks and 8 treatments per block; treatments consisted of 4 ages of cutting (30, 45, 60, and 90 days), with fertilization and without fertilization. The results showed increases of up to 72,000 kg ha-1 year-1 of dry matter (DM) when fertilization was implemented. There was a significant reduction in with an increase in the cutting days (12.70-6.53% protein). Fiber increased (48.79-72.99% NDF) when fertilization treatments were included and cutting days increased. The elements that were included in fertilization (N, P, K) showed a higher foliar content and also presented a reduction in foliar content with growth of the plant. Treatments with fertilization showed a nutrient uptake increase for all the elements up to 60 days, where a reduction in uptake capacity was observed. King grass is a plant with a high nutrient uptake capacity and, therefore, with high biomass and nutrient production. This is an advantage since it can be used in multiple applications, such as animal feed, biofuel production, and as a substrate for biodigestion, among others.

Lipomatosis intestinal: reporte de dos casos / Intestinal lipomatosis: Report of two cases

Resumen La lipomatosis intestinal es un término utilizado para describir una proliferación de tejido graso, histológicamente normal, en el tracto gastrointestinal. Se caracteriza por la presencia de tumores benignos, asintomáticos, de crecimiento lento y de origen subepitelial. En raras ocasiones, estos tumores pueden ser sintomáticos y asociarse a complicaciones importantes como obstrucción y sangrado gastrointestinal. Así, la lipomatosis intestinal es una patología rara, con pocos casos reportados en la literatura y ninguno registrado en Colombia. A continuación, presentamos dos casos evaluados y seguidos en nuestro servicio de Unión de Cirujanos S.A.S de la Universidad de Caldas. El primero es el caso de un hombre de 70 años con historia de episodios de dolor abdominal tipo cólico, asociados a distensión y diarrea posprandial. La endoscopia digestiva alta inicial reportó múltiples lesiones amarillentas, submucosas, en el duodeno, con diagnóstico histológico de lipomas. En este caso, se realizó una videocápsula endoscópica para determinar la distribución y las características de las lesiones presentes en todo el tracto gastrointestinal, así como para valorar la presencia de complicaciones. El otro caso es el de un hombre de 81 años, quien ingresó a la institución por lipotimia y sangrado rectal. Por tanto, se realizó una endoscopia alta y una colonoscopia, cuyos resultados fueron normales. Luego de ello, se efectuó una videocápsula endoscópica que mostró lesiones lipomatosas; una de ellas, con sangrado y angiodisplasia al lado. Estas lesiones se manejaron mediante enteroscopia de doble balón y terapia con argón plasma.

Abstract Intestinal lipomatosis is a proliferation of histologically normal fatty tissue in the gastrointestinal tract. It is characterized benign, asymptomatic, slow growing tumors with sub-epithelial origins. On rare occasions, they become symptomatic and can be associated with major complications such as gastrointestinal obstructions and bleeding. This rare pathology has been reported several times in the literature but not in Colombia prior to this study. We present two cases that were evaluated and followed up at the Unión de Cirujanos SAS and the University of Caldas in Manizales, Colombia. The first is a 70-year-old man who had a history of colic associated with bloating and postprandial diarrhea. The initial upper digestive endoscopy reported multiple yellowish, submucosal lesions in the duodenum. His histological diagnosis was lipomas. In this case, an endoscopic video capsule determined the distribution and characteristics of the lesions throughout the gastrointestinal tract and assessed complications. The second is an 81-year-old man who entered the institution due to lipothymia and rectal bleeding. Upper endoscopy and colonoscopy were normal, but a videocapsule endoscopy showed lipomatous lesions one of which was bleeding and had adjacent angiodysplasia. He was treated with double balloon enteroscopy and Argon plasma therapy.

Nutrient absorption in Tithonia Diversifolia / Absorción de Nutrientes en Tithonia diversifolia / Absorção de nutrientes em Tithonia diversifolia

Abstract Tithonia diversifolia is a robust shrub that has high ecological plasticity and adaptability, high capacity of nutrient absorption and high nutrient contents. These characteristics make Tithonia diversifolia be considered as a multi-purpose plant, such as for animal feed, soil decontamination and soil restorer. Likewise, it is a plant with high ecological plasticity and adaptability. The study of the nutrient absorption and the fertilization represent an important advance in the development of productive systems focused on maximizing forage productivity, which guarantees the soil sustainability. Based on this, a split plot design was conducted to study the effects of different levels of fertilization. The results showed an impact of the elements and levels of fertilization on the nutrient absorption capacity, finding that the elements that were incorporated in the fertilization increase the foliar contents and the nutrient absorption in the plant.

Resumen Tithonia diversifolia es un arbusto robusto que tiene alta plasticidad ecológica y adaptabilidad, alta capacidad de absorción de nutrientes y alto contenido de nutrientes. Estas características hacen que Tithonia diversifolia sea considerada una planta multipropósito, tanto para alimento animal como para descontaminación y restauración del suelo. El estudio de la absorción de nutrientes y la fertilización representa un avance importante en el desarrollo de sistemas productivos enfocados en la maximización de la productividad de forraje, lo cual garantiza la sostenibilidad del suelo. Con base en esto, se llevó a cabo un diseño experimental de parcelas divididas con el fin de estudiar los efectos de diferentes niveles de fertilización. Los resultados mostraron un impacto de los elementos y los niveles de fertilización en la capacidad de absorción de nutrientes; se encontró que los elementos que fueron incorporados en la fertilización incrementan los contenidos foliares y la absorción de nutrientes en la planta.

Resumo Tithonia diversifolia é um arbusto robusto que tem alta plasticidade ecológica e adaptabilidade, alta capacidade de absorção de nutrientes e alto conteúdo de nutrientes. Estas características fazem com que Tithonia diversifolia seja considerada uma planta multipropósito, tanto para alimento animal como para descontaminação e restauração do solo. O estudo da absorção de nutrientes e da fertilização representa um avance importante no desenvolvimento de sistemas produtivos enfocados na otimizacao da produtividade de forragem, o que garante a sustentabilidade do solo. Baseado em esto, se realizou um desenho experimental de parcelas divididas com a finalidade de estudar os efeitos de diferentes níveis de fertilização. Os resultados mostraram um impacto dos elementos e dos níveis de fertilização na capacidade de absorção de nutrientes. Se encontrou que os elementos que foram incorporados na fertilização incrementam os conteúdos foliares e a absorção de nutrientes na planta.

Class II Cytochrome P450 Reductase Governs the Biosynthesis of Alkaloids.

Expansion of the biosynthesis of plant specialized metabolites notably results from the massive recruitment of cytochrome P450s that catalyze multiple types of conversion of biosynthetic intermediates. For catalysis, P450s require a two-electron transfer catalyzed by shared cytochrome P450 oxidoreductases (CPRs), making these auxiliary proteins an essential component of specialized metabolism. CPR isoforms usually group into two distinct classes with different proposed roles, namely involvement in primary and basal specialized metabolisms for class I and inducible specialized metabolism for class II. By studying the role of CPRs in the biosynthesis of monoterpene indole alkaloids, we provide compelling evidence of an operational specialization of CPR isoforms in Catharanthus roseus (Madagascar periwinkle). Global analyses of gene expression correlation combined with transcript localization in specific leaf tissues and gene-silencing experiments of both classes of CPR all point to the strict requirement of class II CPRs for monoterpene indole alkaloid biosynthesis with a minimal or null role of class I. Direct assays of interaction and reduction of P450s in vitro, however, showed that both classes of CPR performed equally well. Such high specialization of class II CPRs in planta highlights the evolutionary strategy that ensures an efficient reduction of P450s in specialized metabolism.

Characterization of a second secologanin synthase isoform producing both secologanin and secoxyloganin allows enhanced de novo assembly of a Catharanthus roseus transcriptome.

BACKGROUND: Transcriptome sequencing offers a great resource for the study of non-model plants such as Catharanthus roseus, which produces valuable monoterpenoid indole alkaloids (MIAs) via a complex biosynthetic pathway whose characterization is still undergoing. Transcriptome databases dedicated to this plant were recently developed by several consortia to uncover new biosynthetic genes. However, the identification of missing steps in MIA biosynthesis based on these large datasets may be limited by the erroneous assembly of close transcripts and isoforms, even with the multiple available transcriptomes. RESULTS: Secologanin synthases (SLS) are P450 enzymes that catalyze an unusual ring-opening reaction of loganin in the biosynthesis of the MIA precursor secologanin. We report here the identification and characterization in C. roseus of a new isoform of SLS, SLS2, sharing 97 % nucleotide sequence identity with the previously characterized SLS1. We also discovered that both isoforms further oxidize secologanin into secoxyloganin. SLS2 had however a different expression profile, being the major isoform in aerial organs that constitute the main site of MIA accumulation. Unfortunately, we were unable to find a current C. roseus transcriptome database containing simultaneously well reconstructed sequences of SLS isoforms and accurate expression levels. After a pair of close mRNA encoding tabersonine 16-hydroxylase (T16H1 and T16H2), this is the second example of improperly assembled transcripts from the MIA pathway in the public transcriptome databases. To construct a more complete transcriptome resource for C. roseus, we re-processed previously published transcriptome data by combining new single assemblies. Care was particularly taken during clustering and filtering steps to remove redundant contigs but not transcripts encoding potential isoforms by monitoring quality reconstruction of MIA genes and specific SLS and T16H isoforms. The new consensus transcriptome allowed a precise estimation of abundance of SLS and T16H isoforms, similar to qPCR measurements. CONCLUSIONS: The C. roseus consensus transcriptome can now be used for characterization of new genes of the MIA pathway. Furthermore, additional isoforms of genes encoding distinct MIA biosynthetic enzymes isoforms could be predicted suggesting the existence of a higher level of complexity in the synthesis of MIA, raising the question of the evolutionary events behind what seems like redundancy.

An acidic phospholipase A2 with antibacterial activity from Porthidium nasutum snake venom.

Snake venoms are complex mixtures of proteins among which both basic and acidic phospholipases A(2) (PLA(2)s) can be found. Basic PLA(2)s are usually responsible for major toxic effects induced by snake venoms, while acidic PLA(2)s tend to have a lower toxicity. A novel PLA(2), here named PnPLA(2), was purified from the venom of Porthidium nasutum by means of RP-HPLC on a C18 column. PnPLA(2) is an acidic protein with a pI of 4.6, which migrates as a single band under both non-reducing and reducing conditions in SDS-PAGE. PnPLA(2) had a molecular mass of 15,802.6 Da, determined by ESI-MS. Three tryptic peptides of this protein were characterized by HPLC-nESI-MS/MS, and N-terminal sequencing by direct Edman degradation showing homology to other acidic PLA(2)s from viperid venoms. PnPLA(2) displayed indirect hemolytic activity in agarose erythrocyte-egg yolk gels and bactericidal activity against Staphylococcus aureus in a dose-dependent manner, with a MIC and MBC of 32 µg/mL. In addition, PnPLA(2) showed a potent inhibitory effect on platelet aggregation with doses up to 40 µg/mL. This acidic PLA(2), in contrast to basic enzymes isolated from other viperid snake venoms, was not cytotoxic to murine skeletal muscle myoblasts C(2)C(12). This is the first report on a bactericidal protein of Porthidium nasutum venom.

Efectos inhibitorios de compuestos fenólicos de plantas sobre la actividad enzimática y citotoxica inducida por una fosfolipasa A2 de veneno de serpiente / Inhibitory effects of plant phenolic compounds on enzymatic and cytotoxic activities induced by a snake venom phospholipase A2

Los compuestos fenólicos han mostrado inhibir los efectos tóxicos inducidos por proteínas de veneno de serpiente. En éste trabajo, nosotros demostramos que el ácido gálico, el ácido ferúlico, el ácido cafeico, el propilgalato y el epigalocatequingalato inhiben la actividad enzimática de una fosfolipasa A2 (PLA2) usando yema de huevo como sustrato. Los valores de IC50 están entre 0,38 – 3,93 mM. Los compuestos mencionados también inhiben la actividad enzimática cuando un sustrato sintético es usado. Además, estos compuestos polifenólicos disminuyen el efecto citotóxico inducido por la fosfolipasa A2 miotóxica, el epigalocatequingalato exhibe la mejor capacidad inhibitoria con 90,92 ± 0,82%, mientras que el ácido ferúlico muestra la menor actividad inhibitoria con 30,96 ± 1,42%. Con el fin de determinar los posibles mecanismos de acción de los compuestos fenólicos, realizamos estudios de modelamiento molecular. Todos los polifenoles muestran puentes de hidrogeno con el sitio activo de la enzima; además el epigalocatequingalato presenta una unión más fuerte con la PLA2 que los otros compuestos. Adicionalmente, un análisis preliminar de relación estructura actividad muestra una correlación entre los valores de IC50 y el área superficial polar topológica (p = 0,0491, r = -0,8079 (-0,9878 a -0,2593)), la cual indica el área superficial requerida por cada molécula para unirse a la enzima. Además, nuestros resultados muestran al propilgalato y el epigalocatequingalato como dos nuevos productos naturales con potencial antimiotóxico. La aplicación tópica de estos polifenoles en el sitio de mordedura podría llevar a la prevención de la miotoxicidad; sin embargo, posteriores investigaciones in vivo serían necesarias para confirmar los resultados in vitro.

[Biomass recovery through secondary succession in the Cordillera Central de los Andes, Colombia]. / Recuperación de la biomasa mediante la sucesión secundaria, Cordillera Central de los Andes, Colombia.

Estimations on biomass recovery rates by secondary tropical forests are needed to understand the complex tropical succession, and their importance on CO2 capture, to offset the warming of the planet. We conducted the study in the Porce River Canyon between 550 and 1 700m.a.s.l. covering tropical and premontane moist belts. We established 33 temporary plots of 50m x 20m in secondary forests, including fallows to succesional forests, and ranging between 3 and 36 years old; we measured the diameter at breast height (D) of all woody plants with D > or = 5cm. In each one of these plots we established five 10m x 10m subplots, in which we measured the diameter betweem 1cm < or = D < 5cm of all woody plants. We estimated the biomass of pastures by harvesting 54 plots of 2m x 2m, and of shrubs in the fallows by harvesting the biomass in 18 plots of 5m x 2m. We modeled Bav (above ground live biomass of woody plants) and Brg (coarse root biomass) as a function of succesional age (t) with the growth model of von Bertalanffy, using 247t/ha and 66t/ha as asymptote, respectively. Besides, we modeled the ratios brg/bav = f(D) and Brg/Bav = f(t). The model estimated that 87 years are required to recover the existing Bav of primary forests through secondary succession, and 217 years for the Brg of the primary forest. The maximum instantaneous growth rate of the Bav was 6.95 t/ha/yr at age 10. The maximum average growth rate of the Bav was 6.26 t/ha/yr at age 17. The weighted average of the absolute growth rate of the Bav reached 4.57t/ha/yr and the relative growth rate 10% annually. The ratio brg/bav decreases with increasing D. The ratio Brg/Bav initially increases very rapidly until age 5 (25%), then decreases to reach 25 years (18%) and increases afterwards until the ratio reaches the asymptote (26.7%).

Recuperación de la biomasa mediante la sucesión secundaria, Cordillera Central de los Andes, Colombia

Biomass recovery through secondary succession in the Cordillera Central de los Andes, Colombia. Estimations on biomass recovery rates by secondary tropical forests are needed to understand the complex tropical succession, and their importance on CO2 capture, to offset the warming of the planet. We conducted the study in the Porce River Canyon between 550 and 1 700m.a.s.l. covering tropical and premontane moist belts. We established 33 temporary plots of 50mx20m in secondary forests, including fallows to succesional forests, and ranging between 3 and 36 years old; we measured the diameter at breast height (D) of all woody plants with D≥5cm. In each one of these plots we established five 10mx10m subplots, in which we measured the diameter betweem 1cm≤D<5cm of all woody plants. We estimated the biomass of pastures by harvesting 54 plots of 2mx2m, and of shrubs in the fallows by harvesting the biomass in 18 plots of 5mx2m. We modeled Bav (above ground live biomass of woody plants) and Brg (coarse root biomass) as a function of succesional age (t) with the growth model of von Bertalanffy, using 247t/ha and 66t/ha as asymptote, respectively. Besides, we modeled the ratios brg/bav=f(D) and Brg/Bav=f(t). The model estimated that 87 years are required to recover the existing Bav of primary forests through secondary succession, and 217 years for the Brg of the primary forest. The maximum instantaneous growth rate of the Bav was 6.95 t/ha/yr at age 10. The maximum average growth rate of the Bav was 6.26 t/ha/yr at age 17. The weighted average of the absolute growth rate of the Bav reached 4.57t/ha/yr and the relative growth rate 10% annually. The ratio brg/bav decreases with increasing D. The ratio Brg/Bav initially increases very rapidly until age 5 (25%), then decreases to reach 25 years (18%) and increases afterwards until the ratio reaches the asymptote (26.7%). Rev. Biol. Trop. 59 (3):1337-1358. Epub 2011 September 01.

En bosques secundarios premontanos tropicales es escasa la información sobre el tiempo requerido para recuperar la biomasa aérea y subterránea de los primarios. Por lo tanto, establecimos 33 parcelas de 0.1ha entre 550 y 1 700m en bosques secundarios que cubrían estadios serales de 3 a 36 años y estimamos la biomasa de las plantas y de las raíces gruesas con ecuaciones locales. Así como la biomasa de las pasturas y barbechos por cosecha de 54 y 18 parcelas, respectivamente. También se calculó la edad de las parcelas con informantes locales, sensores remotos, C14 y anillos de crecimiento. En cada parcela medimos la biomasa aérea viva por hectárea (Bav) y la de las raíces gruesas por hectárea (Brg). Modelamos la Bav y Brg en función de la edad mediante la ecuación de von Bertalanfy con asíntotas de 247t/ha (Bav) y 66t/ha (Brg) resultantes de la medición de 33 parcelas de 0.1ha en los bosques primarios. Con la razón Brg/Bav=f(t) estimamos 87 años para que los bosques secundarios recuperen la Bav existente en los primarios y 217 años para recuperar la Brg. La tasa máxima de crecimiento instantáneo de la Bav fue 6.95t/ha/año a los 10 años y la tasa media máxima de crecimiento 6.26t/ha/año a los 17 años. La media ponderada de la tasa de crecimiento absoluto de la Bav alcanzó 4.57t/ha/año y la relativa 10% anual. La razón Brg/Bav inicialmente aumenta muy aceleradamente hasta 4-5 años (25%), luego disminuye hasta 25 años (18%) para luego incrementar hasta 26.7.

Actividad inhibitoria de Murraya paniculata contra fosfolipasas A2 miotóxicas / Inhibitory activity of Murraya paniculata against myotoxic phospholipases A2

Los envenenamientos causados por serpientes del género Bothrops constituyen un problema de salud en algunos países tropicales, no solamente por la mortalidad sino por el alto porcentaje de incapacidades físicas que ocasionan. Las fosfolipasas A₂(PLA₂) son componentes abundantes en los venenos bothrópicos y crotálicos, y se caracterizan por inducir daños irreversibles en el músculo. Considerando el aporte de las plantas en el tratamiento ofídico, este trabajo se enfocó en la búsqueda de inhibidores de las fosfolipasas, bien sea como alternativa terapéutica o como coadyuvante del tratamiento ofídico. De un tamizaje de 37 especies de plantas vasculares y briofitos contra las PLA₂, se seleccionó la especie Murraya paniculata por su promisoria actividad preliminar. Se siguió con el fraccionamiento biodirigido por la evaluación de la actividad inhibitoria de la PLA₂ y de la fracción con mayor actividad inhibitoria, por medio de cromatografía de gases acoplada a masas se identificaron metabolitos responsables de la actividad, para proponer un modelo de inhibición enzima-sustrato. Los resultados de este proyecto pretenden contribuir en la búsqueda de moléculas inhibitorias de las PLA₂en plantas vasculares, como la especie M. paniculata, que podrían llegar a ser una herramienta en el desarrollo de nuevos productos para mejorar el pronóstico del accidente ofídico.

Envenomation by snakes of the genus Bothrops are a health problem in some tropical countries, not only because of the mortality but given the high percentage of physical disabilities it causes. Phospholipases A2 (PLA2) are abundant constituents in bothropic and crotalic venoms, characterized by inducing irreversible muscle damage. Due to the contribution of plants in the snakebite treatment, we focus on the search of phospholipases inhibitors, either as an alternative treatment or as a therapeutic adjuvant of the current treatments. From a screening of 37 vascular plants and bryophytes in the search for such inhibitors, the species Murraya paniculata was selected due to its promising preliminary activity. Starting from this point further work consisting of bioassay-guided fractionation followed by the evaluation of the inhibitory activity of PLA2, the metabolites responsible for the activity were detected through a tandem of gas chromatography - mass spectrometry in the most active fraction, to propose substrate-enzyme inhibition model. The aims of this project was the search of promising compounds with an inhibitory activity of the PLA2 in vascular plants such as M. paniculata, that could become tools for developing new products to improve the prognosis of the snakebite.

Leucemia de Células dendríticas: reporte de un caso / Dentritic Cell Leukemia: A Case Report

Recientemente, se han reportado varios casos de leucemia plasmocitoide de céluals dendriticas, caracterizados por la expresión de CD4 y CD56, en ausencia de cualquier marcador específico para leucemias mieloides, linfoides B Y t o leucemia de células NK. La ontogenia de estos trastornos no es clara y existe debate alrededor de su relación con precursores mielomonocíticos o de células NK. Su prevalencia se estima en menos del 1 por ciento y puede indentificarse utilizando un inmunofenotipo definido por la presencia de CD45, CD4+,CD56+, CD116, CD123 (receptor de IL-3a), HLA-DR+,CD45RA+,CD45RO,BDCA-2+(CD303+),BDCA-4+(CD304+), Y POR EL ilt-3+.A pesar de que este tipo de leucemia parece ser sensible a la quimioterapia, con frecuencia se encuentra un curso clínico adverso con supervivencia global reducida. Se presenta el primer caso de leucemia plasmocitoide de células dentríticas diagnosticado y tratado en el Instituto Nacional de cancerología de Colombia.

Cellular trafficking of phospholamban and formation of functional sarcoplasmic reticulum during myocyte differentiation.

Phospholamban (PLB) associates with the Ca(2+)-ATPase in sarcoplasmic reticulum (SR) membranes to permit the modulation of contraction in response to beta-adrenergic signaling. To understand how coordinated changes in the abundance and intracellular trafficking of PLB and the Ca(2+)-ATPase contribute to the maturation of functional muscle, we measured changes in abundance, location, and turnover of endogenous and tagged proteins in myoblasts and during their differentiation. We found that PLB is constitutively expressed in both myoblasts and differentiated myotubes, whereas abundance increases of the Ca(2+)-ATPase coincide with the formation of differentiated myotubes. We observed that PLB is primarily present in highly mobile vesicular structures outside the endoplasmic reticulum, irrespective of the expression of the Ca(2+)-ATPase, indicating that PLB targeting is regulated through vesicle trafficking. Moreover, using pulse-chase methods, we observed that in myoblasts, PLB is trafficked through directed transport through the Golgi to the plasma membrane before endosome-mediated internalization. The observed trafficking of PLB to the plasma membrane suggests an important role for PLB during muscle differentiation, which is distinct from its previously recognized role in the regulation of the Ca(2+)-ATPase.

The charged milieu: a major player in fertilization reactions.

In previous studies, we have found that negatively charged, but not uncharged, amino acids and sugars block sea urchin fertilization. These studies were developed from modeling work in non-living systems using derivatized agarose beads that suggested that charge-charge bonding may control at least some adhesive interactions. In the present study, the effects of positively charged, negatively charged and uncharged molecules were examined in the sea urchin sperm-egg system in over 300 individual trials. The results indicate that depending on the specific molecules utilized, both sperm and egg are exquisitely sensitive to charged but not uncharged molecules and to pH changes in sea water caused by some of the charged molecules. It is shown that egg activation, as well as sperm motility and sperm-egg interactions, can be affected by charged molecules. One compound, fructose-1-phosphate blocked fertilization in S. purpuratus sea urchins but not in Lytechinus pictus sea urchins. These findings indicate that charge alone cannot explain all the results. In this case, the presence of a ketone instead of an aldehyde group indicates that species-specific components may control fertilization reactions. The present study is a comprehensive survey of the effects of charge, pH and molecular structure on the fertilization activation continuum in a model system of sea urchins.

Regulation of cysteine dioxygenase and gamma-glutamylcysteine synthetase is associated with hepatic cysteine level.

Two hepatic enzymes, cysteine dioxygenase (CDO) and gamma-glutamylcysteine synthetase (GCS), play important regulatory roles in the response of cysteine metabolism to changes in dietary sulfur amino acid or protein levels. To examine the time-course of changes in CDO and GCS activities, CDO and GCS-catalytic or heavy subunit protein and mRNA levels, and cysteine and glutathione levels, we adapted rats to either a low protein (LP) or high protein (HP) diet, switched them to the opposite diet, and followed these parameters over 6 days. Hepatic CDO activity and amount, but not mRNA level, increased in response to higher protein intake; the t(1/2) of change for CDO activity or protein level was 22 h for rats switched from a LP to a HP diet and 8 h for rats switched from a HP to a LP diet, suggesting that the HP diet decreased turnover of CDO. Hepatic GCS activity, catalytic subunit amount and mRNA level decreased in response to a higher protein intake. GCS catalytic subunit level changed with a similar t(1/2) for both groups, but the change in GCS activity in rats switched from a LP diet to a HP diet was faster (approximately 16h) than for rats switched from a HP to a LP diet (approximately 74h). Hepatic cysteine and glutathione levels reached new steady states within 12 h (LP to HP) or 24 h (HP to LP). CDO activity appeared to be regulated at the level of protein, probably by diminished turnover of CDO in response to higher protein intake or cysteine level, whereas GCS activity appeared to be regulated both at the level of mRNA and activity state in response to the change in cysteine or protein availability. These findings support a role of cysteine concentration as a mediator of its own metabolism, favoring catabolism when cysteine is high and glutathione synthesis when cysteine is low.

The ubiquitin-proteasome system is responsible for cysteine-responsive regulation of cysteine dioxygenase concentration in liver.

Hepatic cysteine dioxygenase (CDO) activity is a critical regulator of cellular cysteine concentration and availability of cysteine for anabolic processes and is markedly higher in animals fed diets containing excess sulfur amino acids compared with those fed levels at or below the requirement. Rat hepatocytes responded to a deficiency or excess of cysteine in the culture medium with a decrease or increase in CDO level but no change in CDO mRNA level. The cysteine analog, cysteamine, but not cysteine metabolites or thiol reagents, was also effective in increasing CDO. Inhibitors of the 26S proteasome blocked CDO degradation in cysteine-deficient cells but had little or no effect on CDO concentration in hepatocytes cultured with excess cysteine. High-molecular-mass CDO-ubiquitin conjugates were observed in cells cultured in cysteine-deficient medium, whether or not proteasome inhibitor was present, but these CDO-ubiquitin conjugates were not observed in cells cultured in cysteine-supplemented medium with or without proteasome inhibitor. Similar results were observed for degradation of recombinant CDO expressed in human heptocarcinoma cells cultured in cysteine-deficient or cysteine-supplemented medium. CDO is an example of a mammalian enzyme that is robustly regulated via its substrate, with the presence of substrate blocking the ubiquitination of CDO and, hence, the targeting of CDO for proteasomal degradation. This regulation occurs in primary hepatocytes in a manner that corresponds with changes observed in intact animals.

Enzymes and metabolites of cysteine metabolism in nonhepatic tissues of rats show little response to changes in dietary protein or sulfur amino acid levels.

In liver, cysteine dioxygenase (CDO), cysteinesulfinate decarboxylase (CSD), and gamma-glutamylcysteine synthetase (GCS) play important regulatory roles in the metabolism of cysteine to sulfate, taurine and glutathione. Because glutathione is released by the liver and degraded by peripheral tissues that express gamma-glutamyl transpeptidase, some peripheral tissues may be exposed to relatively high concentrations of cysteine. Rats were fed diets that contained low, moderate or high concentrations of protein or supplemental cysteine or methionine for 2 wk, and CDO, CSD and GCS activities, concentrations and mRNA levels and the concentrations of cysteine, taurine and glutathione were measured in liver, kidney, lung and brain. All three enzymes in liver responded to the differences in dietary protein or sulfur amino acid levels, but only CSD in kidney and none of the three enzymes in lung and brain responded. Renal CSD activity was twice as much in rats fed the low protein diet as in rats fed the other diets. Changes in renal CSD activity were correlated with changes in CSD concentration. Some significant differences in cysteine concentration in kidney and lung and glutathione and taurine concentrations in kidney were observed, with higher concentrations in rats fed higher levels of protein or sulfur amino acids. In liver, the changes in cysteine level were consistent with cysteine-mediated regulation of hepatic CDO activity, and changes in taurine level were consistent with predicted changes in cysteine catabolism due to the changes in cysteine concentration and CDO activity. Changes in renal and lung cysteine, taurine or glutathione concentrations were not associated with a similar pattern of change in CDO, CSD or GCS activity. Overall, the results confirm the importance of the liver in the maintenance of cysteine homeostasis.