Discovery of the ARP2 protein as a determining molecule in tumor cell death

Cancer is a multifactorial disease that constitutes a serious public health problem worldwide. Prostate cancer advanced stages are associated with the development of androgen-independent tumors and an apoptosis-resistant phenotype that progresses to metastasis. By studying androgen-independent lymphoid nodule carcinoma of the prostate (LNCaP) cells induced to apoptosis by serum elimination, we identified the activation of a non-selective cationic channel of 23pS conductance that promotes incoming Ca2+ currents, as well as apoptosis final stages. arp2cDNA was isolated and identified to be of the same cell type, and mRNA was expressed in Xenopus laevis oocytes, which was found to be associated with the activation of incoming Ca2+ currents and induction to apoptosis. cDNA, which encodes the ARP2 protein, was overexpressed in LNCaP cells and Chinese hamster ovary cells, which induced apoptosis. Our evidence suggests that protein ARP2 overexpression and transit to the cell membrane allows an increased Ca2+ incoming current that initiates the apoptosis process in epithelial-type cells whose phenotype shows resistance to programmed cell death.

Microvascular anatomy of olfactory and accessory olfactory (vomeronasal) organs in adult Xenopus laevis: scanning electron microscopy of vascular corrosion casts / Anatomía microvascular de órganos olfatorios y accesorios (vomeronasal) en adultos Xenopus laevis: microscopía electrónica de barrido de moldes de corrosión vascular

Microvascular anatomy and histomorphology of olfactory and vomeronasal organs in adult Xenopus laevis Daudin were studied by scanning electron microscopy of vascular corrosion casts and paraplast embedded stained serial tissue sections. Results show that the arterial supply is bilaterally by terminal arterioles of the medial branch of the nasal artery and by the palatal artery. Arterioles give rise to a capillary meshwork characteristic for respiratory surfaces in principal chambers and in dorsal and caudal areas of middle chambers. Anterior and inferior areas of the middle chambers own a distinctly different capillary network with conspicuous short capillary loops. Loops have a dilated tip and extend in acute angles towards the chamber lumen. The vomeronasal organ (VNO) locates beneath the olfactory organ. It has a medial to lateral extension and attaches with its caudal circumference to the medial nasal glands. Its capillary bed displays rectangular meshes which preferentially orientate along the long axis of the VNO. Locally, capillaries form short hairpin-like or strongly twisted loops with dilated tips which point towards the lumen of the VNO. These capillaries slow-down blood velocity and may lead to an increased exchange of oxygen, nutrients and water-borne odorants in the middle chambers and of pheromones in the VNO. In the latter vascular structures are present which might serve as a vascular pump.

Se estudiaron la anatomía microvascular e histomorfología de los órganos olfatorios y vomeronasales de Xenopus laevis Daudin adultos, mediante microscopía electrónica de barrido de moldes de corrosión vascular y secciones de tejido seriadas, teñidas e incluídas en paraplast. Los resultados muestran que el suministro arterial es bilateral por arteriolas terminales de la rama medial de la arteria nasal y por la arteria palatina. Las arteriolas dan lugar a un lecho capilar característico de las superficies respiratorias en las cámaras principales y en las áreas dorsal y caudal de las cámaras intermedias. Las áreas anterior e inferior de las cámaras centrales poseen una red capilar significativamente diferente con llamativos bucles capilares cortos. Los bucles tienen una punta dilatada y se extienden en ángulos agudos hacia la luz de la cámara. El órgano vomeronasal (VNO) se ubica debajo del órgano olfatorio. Se extiende de medial a lateral y se une con su circunferencia caudal a las glándulas nasales mediales. El lecho capilar muestra mallas rectangulares que se orientan preferentemente a lo largo del eje longitudinal del VNO. Localmente, los capilares forman bucles cortos en forma de horquilla o fuertemente retorcidos con puntas dilatadas que apuntan hacia la luz del VNO. Estos capilares ralentizan la velocidad de la sangre y pueden conducir a un mayor intercambio de oxígeno, nutrientes y odorizantes, a base de agua en las cámaras intermedias y de feromonas, en el VNO. En este último, están presentes estructuras vasculares que podrían servir como una bomba vascular.

First parasitological study of the African clawed frog (Xenopus laevis, Amphibia) in Chile / Primeiro estudo parasitológico em rã com garras Africano (Xenopus laevis, Anfibia) no Chile

Abstract Introduced species can arrive into new territories with parasites; however, these species are expected to face lower parasite richness than in their original regions. Both introduced hosts and parasites can affect native fauna. Since their release into the wild in Chile following laboratory use, Xenopus laevis Daudin, 1802 has widely spread throughout central Chile. The only pathogen described on the host is the fungus Batrachochytrium dendrobatidis Longcore, Pessier, Nichols, 1999; thus, this is the first parasitological study of this species in Chile. In 10 localities in central Chile, 179 specimens of X. laevis were captured and examined for parasites in the gastrointestinal tube, cavities, lungs, liver, and skin. Only nine specimens of the genus Contracaecum Railliet, Henry, 1912 were found in six specimens of X. laevis from a private dam in La Patagua. It is likely that these parasites originated from species of native birds. This is the first record of Contracaecum sp. in Chilean amphibians.

Resumo Espécies exóticas podem se introduzir em um novo território com seus parasitas, porém nesses casos, a riqueza parasitária seria menor. Contudo, hospedeiros exóticos e seus parasitas associados podem afetar a fauna nativa. Depois de ser dispensado do uso em laboratórios e solto em ambientes naturais, Xenopus laevis Daudin, 1802 tem se espalhado massivamente no Chile central. O único patógeno descrito para este anuro é o fungo Batrachochytrium dendrobatidis Longcore, Pessier, Nichols, 1999. O presente estudo constitui a primeira pesquisa parasitológica realizada nesta espécie de rã introduzida no Chile. Em 10 localidades do Chile central, foram capturados 179 espécimes de X. laevis que foram examinadas em busca de parasitos dentro tubo digestivo, cavidades corporais, pulmões, fígado e pele. Nove espécimes do gênero Contracaecum Railliet, Henry, 1912 foram encontrados em seis espécimes de X. laevis de uma barragem em La Patagua. É provável que a origem destes parasitas sejam espécies de aves nativas. Este é o primeiro relato de Contracaecum sp. em anuros do Chile.

Consumption and expenditure on food prepared away from home among Mexican adults in 2006 / Consumo y gasto en alimentos preparados fuera de casa en adultos mexicanos en 2006

Objective. To describe food expenditure and consumption of foods prepared away from home among Mexican adults. Materials and methods. Data were from 45 241 adult participants in the National Health and Nutrition Survey 2006, a nationally-representative, cross-sectional survey of Mexican households. Descriptive statistics and multivariable linear and logistic regression were used to assess the relationship between location of residence, educational attainment, socioeconomic status and the following: 1) expenditure on all food and at restaurants, and 2) frequency of consumption of comida corrida or restaurant food and street food. Results. Food expenditure and consumption of food prepared away from home were positively associated with socioeconomic status, educational attainment, and urban vs. rural residence (p<0.001 for all relationships in bivariate analyses). Conclusions. Consumption of food prepared outside home may be an important part of the diet among urban Mexican adults and those with high socioeconomic status and educational attainment.

Objetivo. Describir los gastos en alimentos y el consumo de alimentos preparados fuera de casa en población mexicana. Material y métodos. Los datos fueron de 45 241 adultos mexicanos en la Encuesta Nacional de Salud y Nutrición de 2006, representativa al nivel nacional. Se utilizaron estadísticas descriptivas y regresión linear y logística para estimar la relación entre el lugar de residencia, el nivel educativo y el nivel socioeconómico, con el gasto en todos los alimentos y en restaurantes, y con la frecuencia de consumo de comida corrida, en restaurantes y de la calle. Resultados. El gasto en alimentos y el consumo de alimentos preparados se asociaron positivamente con el nivel socioeconómico, el nivel educativo y la residencia rural (p<0,001 para todas las relaciones). Conclusiones. El consumo de alimentos preparados puede ser una parte importante de la dieta de los adultos urbanos y de aquéllos con altos niveles socioeconómicos y educativos.

Aparición de melanina como pigmento protector en el encéfalo de xenopus laevis para protegerlo de los efectos de la radiación ultravioleta / Emerging melanin protective pigment in the brain of xenopus laevis to protect from the effects of uv radiation

Con el fin de proteger al organismo de condiciones estresantes, tales como cambios de osmolaridad y de temperatura, además de actuar como pantalla protectora en contra de los rayos ultravioleta (UV). Se ha observado que ciertos anfibios han desarrollado pigmentación en su encéfalo como una posible protección ante el aumento de la radiación UV, causada por el daño en la capa de ozono, la cual estaría alterando al ecosistema. En este trabajo se describe la presencia de pigmentación en el encèfalo de X. laevis durante el desarrollo larvario y su posible función protectora frente a la radiación UV. Para ello, se recolectaron individuos de diferentes estados larvarios, los que fueron obtenidos de distintas localidades de la región de Valparaíso (V región, Chile), para ser procesados con el método corriente H-E y el método de Lillie. En los análisis se pudo evidenciar que la pigmentación correspondía a melanina, que se encontraría en la membrana denominada leptomeninge, la cual recubre al encéfalo y estaría actuando como un filtro protector para evitar daños a nivel del desarrollo en el sistema nervioso de estos anuros. En suma, los rayos UV como agentes deletéreos estarían estimulando la producción de eumelanina en la leptomeninge de estos anfibios, para proteger parte del SNC (encéfalo), como al individuo en sí de posibles alteraciones teratogénicas y/o mutagénicas.

It has been observed that certain amphibians have developed pigmentation in brain as a possible increased protection against UV radiation, caused by damage to the ozone layer, which would alter the ecosystem. In this paper we describe the presence of pigment in the brain of X. laevis during larval development and possible protective function against UV radiation. To do this, we collected individuals at various larval stages, which were obtained from different locations in Valparaiso (V Region, Chile), to be processed with HE and the method of Lillie. In the analysis it was evident that pigmentation corresponded to melanin, which would be in the membrane called leptomeninges, which covers the brain and would be acting as a protective filter to prevent damage to the level of development in the nervous system of these frogs. In addition, UV rays would be deleterious agents stimulating production of eumelanin in the leptomeninges of these amphibians, to protect the CNS (brain), and the individual itself of potential teratogenic or mutagenic alterations.

The expression of melanopsin and clock genes in Xenopus laevis melanophores and their modulation by melatonin

Vertebrates have a central clock and also several peripheral clocks. Light responses might result from the integration of light signals by these clocks. The dermal melanophores of Xenopus laevis have a photoreceptor molecule denominated melanopsin (OPN4x). The mechanisms of the circadian clock involve positive and negative feedback. We hypothesize that these dermal melanophores also present peripheral clock characteristics. Using quantitative PCR, we analyzed the pattern of temporal expression of Opn4x and the clock genes Per1, Per2, Bmal1, and Clock in these cells, subjected to a 14-h light:10-h dark (14L:10D) regime or constant darkness (DD). Also, in view of the physiological role of melatonin in the dermal melanophores of X. laevis, we determined whether melatonin modulates the expression of these clock genes. These genes show a time-dependent expression pattern when these cells are exposed to 14L:10D, which differs from the pattern observed under DD. Cells kept in DD for 5 days exhibited overall increased mRNA expression for Opn4x and Clock, and a lower expression for Per1, Per2, and Bmal1. When the cells were kept in DD for 5 days and treated with melatonin for 1 h, 24 h before extraction, the mRNA levels tended to decrease for Opn4x and Clock, did not change for Bmal1, and increased for Per1 and Per2 at different Zeitgeber times (ZT). Although these data are limited to one-day data collection, and therefore preliminary, we suggest that the dermal melanophores of X. laevis might have some characteristics of a peripheral clock, and that melatonin modulates, to a certain extent, melanopsin and clock gene expression.

Functional characterization and localization of AQP3 in the human colon

Water channels or aquaporins (AQPs) have been identified in a large variety of tissues. Nevertheless, their role in the human gastrointestinal tract, where their action is essential for the reabsorption and secretion of water and electrolytes, is still unclear. The purpose of the present study was to investigate the structure and function of water channels expressed in the human colon. A cDNA fragment of about 420 bp with a 98 percent identity to human AQP3 was amplified from human stomach, small intestine and colon by reverse transcription polymerase chain reaction (RT-PCR) and a transcript of 2.2 kb was expressed more abundantly in colon than in jejunum, ileum and stomach as indicated by Northern blots. Expression of mRNA from the colon of adults and children but not from other gastrointestinal regions in Xenopus oocytes enhanced the osmotic water permeability, and the urea and glycerol transport in a manner sensitive to an antisense AQP3 oligonucleotide, indicating the presence of functional AQP3. Immunocytochemistry and immunofluorescence studies in human colon revealed that the AQP3 protein is restricted to the villus epithelial cells. The immunostaining within these cells was more intense in the apical than in the basolateral membranes. The presence of AQP3 in villus epithelial cells suggests that AQP3 is implicated in water absorption across human colonic surface cells

Bloqueo del ion bario sobre una coriente de potasio rectificadora entrante en ovocitos de rana xenopus laevis / Blockade of barium ions on inward rectifier potassium current study in frog's oocytes

Se caracterizó el efecto del ion bario (Ba²+) sobre un canal de potasio rectificador entrante endógeno (IRK-endo) de ovocitos de Xenopus laevis con la técnica de fijación de voltaje con 2 microelectrodos. Al modificar la [K+] o se provocó un cambio en el potencial de inversión y en el valor de la pendiente de la recta, tal y como se espera para una corriente predominantemente de K+. El bloqueo de IRK-endo por el Ba²+ externo presentó una baja afinidad del canal de dos formas diferentes. Una fue un bloqueo en el pico de corriente independiente de voltaje con una Kd de 0.9 ñ 0.1 mM. En contraste, un segundo efecto se observó al final del pulso (80 ms). Este bloqueo en la corriente es dependiente tanto de tiempo como de voltaje, con una Kd que muestra una dependencia de voltaje monoexponencial (t = 50 ñ 5 mV). Estos resultados sugieren la presencia de dos sitios de unión diferentes para el Ba²+; uno en la parte externa del canal y otro situado dentro del canal Nuestros resultados demuestran la existencia de canal. Nuestros resultados demuestran la existencia de una corriente endógena de K+ en ovocitos de Xenopus laevis con características semejantes a otras corrientes rectificadoras entrantes de K+ encontradas en otras especies

Lectinas solubles beta-galactosídicas / Beta-galactoside solubles lectins

Sobre la base de estudios estructurales y funcionales, las lectinas animales se han clasificado en dos tipos: el Tipo C, caracterizado por su dependencia de los iones de calcio y el Tipo S que no es calciodependiente, sino tioldependiente. Entre estas últimas, se ha estudiado ampliamente el grupo de lectinas S-Lac, que son extraídas con buffers salinos con lactosa, en presencia de tioles. Contituyen una familia de proteínas realcionadas estructuralmente, que contienen una serie de aminoácidos conservados. Se unen específicamente a glicoconjugados complementarios y su biosíntesis y localización son reguladas por el desarrolo. Su rol puede relacionarse con diversas actividades biológicas que poderían variar según el órgano.

Isolation and properties of a multicatalytic proteinase complex from Xenopus laevis skin secretion

A multicatalytic proteinase complex present in the skin secretion of Xenopus laevis was purified and its enzymatic activity towards natural and synthetic peptides was investigated. We identified three activities: i) a C-terminal deamidation enzyme activity which exhibited selectivity for the Asp-Phe-NH2 and Phe-Leu-NH2 motifs of cerulein, minigastrin Leu-enkephalinamide, (des-Tyr1)Leu-enkephalinamide and diaminobenzylthiocyanate-DVDERDVRGFASFL NH2 (DABTCDR8kermit); ii) an endopeptidase activity that cleaves peptide bonds on the carboxyl side of hydrophobic amino acid residues such as Tyr-Gly of LHRH, Ile-Ala of PGLa and Leu-Ala of buccalin; iii) an enzyme activity that cleaves peptide bonds at the dibasic sites of peptides of the dynorphin family. The molecular weight determined by Sephacryl S-400 molecular sieve filtration indicated an Mr about 600 kDa. The activities characterized here exhibit and optimal pH of about 7.4. The activities of the multicatalytic complex were differentially inhibited by the classical inhibitors of proteases

The nicotinic acetylcholine receptor channel in embryonic muscle of xenopus laevis. A program to separate channel types

En este trabajo se emplea la técnica de control de voltaje en pequeños parches de membrana, para evaluar la actividad eléctrica unitaria del receptor-canal nicotínico para acetilcolina (AChR) en miotomos extraídos de embriones de Xenopus laevis. Dos patrones de actividad unitaria del receptor nicotínico para la acetilcolina, se distinguen por su conductancia, 60 y 40 pS. Regularmente, la actividad de ambos tipos de canales aparece en los registros y las complicaciones del análisis cinético estacionario producidas por la mezcla de los dos tipos de actividad se superaron a través del uso del programa de separación "CLASS"

Aeromonas hydrophilia: infección en xenopus leavis / Aeromonas hydrophila: infection in xenopus leavis

Aeromonas hydrophilia causó enfermedad grave en un grupo de 50 Xenopus leavis a las 3 semanas de haber sido trasladados de sus condiciones de laboratorio. Los primeros signos clínicos observados fueron una marcada palidez de la piel, petequias, letargia, anorexia y edemas. La duración de la enfermedad fue aproximadamente de 48 días, durante los cuales 4 animales enfermaron y 10 murieron. Al realizarse estudios bacteriológicos de la piel y del músculo esquelético, se observaron en la necropsia. edemas subcutáneos, hemorragias y ascitis. Se aisló Aeromonas hydrophilia en 14 de los 50 animales estudiados

Single acetylcholine channel currents recorded from xenopus myocytes are influenced by the lass of the patch pipette

El tipo de vidrio que se utiliza para construir las pipetas de "patch clamp" altera las propiedades cinéticas de las corrientes unitarias activadas por acetilcolina en miocitos de Xenopus. En resgistros hechos con pipetas de vidrio duro, la velocidad de apertura disminuye con el transcurso del tiempo. Esta disminución es mas pronunciada para el canal de baja conductancia. Además, los eventos de mayor duración de esta forma del canal desaparecen mas rápidamente que los eventos mas cortos y, como consecuencia, el tiempo abierto del canal se acorta durante el período de registro. Estos efectos no se observan cuando los canales son registrados utilizando pipetas fabricadas con vidrio blando