The Diversity of Gut Microbiota at Weaning Is Altered in Prolactin Receptor-Null Mice.

Maternal milk supports offspring development by providing microbiota, macronutrients, micronutrients, immune factors, and hormones. The hormone prolactin (PRL) is an important milk component with protective effects against metabolic diseases. Because maternal milk regulates microbiota composition and adequate microbiota protect against the development of metabolic diseases, we aimed to investigate whether PRL/PRL receptor signaling regulates gut microbiota composition in newborn mice at weaning. 16SrRNA sequencing of feces and bioinformatics analysis was performed to evaluate gut microbiota in PRL receptor-null mice (Prlr-KO) at weaning (postnatal day 21). The normalized colon and cecal weights were higher and lower, respectively, in the Prlr-KO mice relative to the wild-type mice (Prlr-WT). Relative abundances (Simpson Evenness Index), phylogenetic diversity, and bacterial concentrations were lower in the Prlr-KO mice. Eleven bacteria species out of 470 differed between the Prlr-KO and Prlr-WT mice, with two genera (Anaerotruncus and Lachnospiraceae) related to metabolic disease development being the most common in the Prlr-KO mice. A higher metabolism of terpenoids and polyketides was predicted in the Prlr-KO mice compared to the Prlr-WT mice, and these metabolites had antimicrobial properties and were present in microbe-associated pathogenicity. We concluded that the absence of the PRL receptor altered gut microbiota, resulting in lower abundance and richness, which could contribute to metabolic disease development.

EGFR and p38MAPK Contribute to the Apoptotic Effect of the Recombinant Lectin from Tepary Bean (Phaseolus acutifolius) in Colon Cancer Cells.

Previous works showed that a Tepary bean lectin fraction (TBLF) induced apoptosis on colon cancer cells and inhibited early colonic tumorigenesis. One Tepary bean (TB) lectin was expressed in Pichia pastoris (rTBL-1), exhibiting similarities to one native lectin, where its molecular structure and in silico recognition of cancer-type N-glycoconjugates were confirmed. This work aimed to determine whether rTBL-1 retained its bioactive properties and if its apoptotic effect was related to EGFR pathways by studying its cytotoxic effect on colon cancer cells. Similar apoptotic effects of rTBL-1 with respect to TBLF were observed for cleaved PARP-1 and caspase 3, and cell cycle G0/G1 arrest and decreased S phase were observed for both treatments. Apoptosis induction on SW-480 cells was confirmed by testing HA2X, p53 phosphorylation, nuclear fragmentation, and apoptotic bodies. rTBL-1 increased EGFR phosphorylation but also its degradation by the lysosomal route. Phospho-p38 increased in a concentration- and time-dependent manner, matching apoptotic markers, and STAT1 showed activation after rTBL-1 treatment. The results show that part of the rTBL-1 mechanism of action is related to p38 MAPK signaling. Future work will focus further on the target molecules of this recombinant lectin against colon cancer.

Quantum Dot Labelling of Tepary Bean (Phaseolus acutifolius) Lectins by Microfluidics.

Lectins are bioactive proteins with the ability to recognize cell membrane carbohydrates in a specific way. Diverse plant lectins have shown diagnostic and therapeutic potential against cancer, and their cytotoxicity against transformed cells is mediated through the induction of apoptosis. Previous works have determined the cytotoxic activity of a Tepary bean (Phaseolus acutifolius) lectin fraction (TBLF) and its anti-tumorigenic effect on colon cancer. In this work, lectins from the TBLF were additionally purified by ionic-exchange chromatography. Two peaks with agglutination activity were obtained: one of them was named TBL-IE2 and showed a single protein band in two-dimensional electrophoresis; this one was thus selected for coupling to quantum dot (QD) nanoparticles by microfluidics (TBL-IE2-QD). The microfluidic method led to low sample usage, and resulted in homogeneous complexes, whose visualization was achieved using multiphoton and transmission electron microscopy. The average particle size (380 nm) and the average zeta potential (-18.51 mV) were determined. The cytotoxicity of the TBL-IE2 and TBL-IE2-QD was assayed on HT-29 colon cancer cells, showing no differences between them (p ≤ 0.05), where the LC50 values were 1.0 × 10-3 and 1.7 × 10-3 mg/mL, respectively. The microfluidic technique allowed control of the coupling between the QD and the protein, substantially improving the labelling process, providing a rapid and efficient method that enabled the traceability of lectins. Future studies will focus on the potential use of the QD-labelled lectin to recognize tumor tissues.

Disruption of protein kinase A localization induces acrosomal exocytosis in capacitated mouse sperm.

Protein kinase A (PKA) is a broad-spectrum Ser/Thr kinase involved in the regulation of several cellular activities. Thus, its precise activation relies on being localized at specific subcellular places known as discrete PKA signalosomes. A-Kinase anchoring proteins (AKAPs) form scaffolding assemblies that play a pivotal role in PKA regulation by restricting its activity to specific microdomains. Because one of the first signaling events observed during mammalian sperm capacitation is PKA activation, understanding how PKA activity is restricted in space and time is crucial to decipher the critical steps of sperm capacitation. Here, we demonstrate that the anchoring of PKA to AKAP is not only necessary but also actively regulated during sperm capacitation. However, we find that once capacitated, the release of PKA from AKAP promotes a sudden Ca2+ influx through the sperm-specific Ca2+ channel CatSper, starting a tail-to-head Ca2+ propagation that triggers the acrosome reaction. Three-dimensional super-resolution imaging confirmed a redistribution of PKA within the flagellar structure throughout the capacitation process, which depends on anchoring to AKAP. These results represent a new signaling event that involves CatSper Ca2+ channels in the acrosome reaction, sensitive to PKA stimulation upon release from AKAP.

Sp-tetraKCNG: A novel cyclic nucleotide gated K(+) channel.

The sequence of a novel cGMP-regulated, tetrameric, K(+) selective channel (Sp-tetraKCNG) was discovered in the sea urchin Strongylocentrotus purpuratus. The Sp-tetraKCNG is a single polypeptide made of four KCNG domains similar to voltage-dependent Na(+) and Ca(2+) channels. Each KCNG domain has six transmembrane segments (S1-S6), the ion pore having the K(+) selectivity signature GYGD and a cyclic nucleotide-binding domain (CNBD). This novel channel is evolutionary located between K(+)-selective and voltage-dependent EAG channels and voltage-independent cationic CNG channels. Bilayer reconstitutions demonstrate such a cGMP-regulated K(+) selective channel in sea urchin spermatozoa.

[Dental caries in schoolchildren aged 6-12 years in Navolato, Sinaloa, México: experience, prevalence, severity and treatment needs]. / Caries dental en escolares de 6 a 12 añios de edad en Navolato, Sinaloa, México: experiencia, prevalencia, gravedad y necesidades de tratamiento.

INTRODUCTION: Dental caries is the most prevalent disease in children. OBJECTIVE: To determine the caries experience, prevalence, and severity in temporal and permanent dentition, likewise to establish the significant caries index in schoolchildren of Navolato, Sinaloa, Mexico as well as their treatment needs. MATERIAL AND METHODS: A cross sectional study in 3048 schoolchildren aged 6 to 12 years. All subjects were clinically examined by three examiners (kappa>0.85), according to World Health Organization (WHO) guidelines. Caries detection criteria used were the WHO's criteria and Pitts' lesion d1. RESULTS: The mean age was 8.81 +/- 1.79 years old and 52.2% were girls. In the temporal dentition; the dmft was 4.68 +/- 3.21 and caries prevalence of 90.2% (dmft>3 was 60.8%, and dmft>6 was 27.4%). The overall significant caries index was 10.52 for 6 year olds. The noncavitated lesions (d1) represented 37.1% of the "decay" component. In the permanent dentition; the DMFT was 3.24 +/- 2.72 and caries prevalence was 82.0% (DMFT>3 was 47.8%, and DMFT>6 was 9.0%). The overall significant caries index was 10.87 for 12 year olds. The noncavitated lesions (d1) represented 63.4% of the "decay" component. The majority, 89.6% and 81.1% of children needed at least one restoration of one and two dental surfaces, respectively. Girls were more affected by caries than boys in permanent dentition, but not in temporal dentition. The caries experience increased with age. CONCLUSIONS: We observed high indices of caries, and high prevalence, severity and treatment needs. Noncavitated lesions were higher in permanent dentition that in primary dentition.

Caries dental en escolares de 6 a 12 años de edad en Navolato, Sinaloa, México: experiencia, prevalencia, gravedad y necesidades de tratamiento / Dental caries in schoolchildren aged 6-12 years in Navolato, Sinaloa, México: experience, prevalence, severity and treatment needs

Introducción. La caries dental es una de las enfermedades más prevalentes en los niños. Objetivo. Determinar la experiencia, prevalencia y gravedad de caries en la dentición temporal y permanente; establecer igualmente el índice de caries significativa (SiC, por sus siglas en inglés), en escolares de Navolato, Sinaloa, México, así como sus necesidades de tratamiento. Materiales y métodos. Se realizó un estudio transversal en 3.048 niños de 6 a 12 años de edad. Los sujetos fueron examinados clínicamente por tres examinadores (índice k > 0,85), de acuerdo a los lineamientos establecidos por la Organización Mundial de la Salud (OMS). Para la detección de caries se empleó el criterio de la OMS y la lesión d1 de Pitts. Resultados. La media de edad fue de 8,81±1,79 años y el porcentaje de niñas fue de 52,2 por ciento. En la dentición temporal, el ceod (Promedio de dientes cariados, extraídos/perdidos y obturados) fue de 4,68±3,21 y la prevalencia de caries 90,2 por ciento (ceod > 3 = 60,8 por ciento). El índice significativo de caries fue de 10,52 para los niños de seis años de edad. Las lesiones no cavitadas (d1) representaron 37,1 por ciento del componente "cariados". En la dentición permanente, el índice CPOD (Promedio de dientes permanentes cariados, perdidos y obturados) fue de 3,24±2,72 y la prevalencia de caries, 82 por ciento (CPOD > 3 = 47,8 por ciento). El índice de caries significativa fue de 10,87 para los niños de 12 años. Las lesiones no cavitadas representaron 63,4 por ciento del componente "cariados". Al menos 81,1 por cientode los niños necesitó la restauración de una superficie dental y 89,6 por ciento, de dos. Las niñas presentaron más caries que los niños en su dentición permanente. Se observó que conforme aumentó la edad se incrementó la experiencia de caries.

KATP channels in mouse spermatogenic cells and sperm, and their role in capacitation.

Mammalian sperm must undergo a series of physiological changes after leaving the testis to become competent for fertilization. These changes, collectively known as capacitation, occur in the female reproductive tract where the sperm plasma membrane is modified in terms of its components and ionic permeability. Among other events, mouse sperm capacitation leads to an increase in the intracellular Ca(2+) and pH as well as to a hyperpolarization of the membrane potential. It is well known that ion channels play a crucial role in these events, though the molecular identity of the particular channels involved in capacitation is poorly defined. In the present work, we report the identification and potential functional role of K(ATP) channels in mouse spermatogenic cells and sperm. By using whole-cell patch clamp recordings in mouse spermatogenic cells, we found K(+) inwardly rectifying (K(ir)) currents that are sensitive to Ba(2+), glucose and the sulfonylureas (tolbutamide and glibenclamide) that block K(ATP) channels. The presence of these channels was confirmed using inhibitors of the ATP synthesis and K(ATP) channel activators. Furthermore, RT-PCR assays allowed us to detect transcripts for the K(ATP) subunits SUR1, SUR2, K(ir)6.1 and K(ir)6.2 in total RNA from elongated spermatids. In addition, immunoconfocal microscopy revealed the presence of these K(ATP) subunits in mouse spermatogenic cells and sperm. Notably, incubation of sperm with tolbutamide during capacitation abolished hyperpolarization and significantly decreased the percentage of AR in a dose-dependent fashion. Together, our results provide evidence for the presence of K(ATP) channels in mouse spermatogenic cells and sperm and disclose the contribution of these channels to the capacitation-associated hyperpolarization.

Diseno de una planta para la transformacion y tratamiento industrial de maderas.Estudio tecnologico y economico

Contenido: Cap.1 Descripcion sumaria del proyecto Cap.2 Analisis de los recursos forestales.Introduccion.Recursos forestales en Bolivia.Riqueza forestal por departamentos.Zonas de aprovechamiento.Explotacion de especies forestales.Potencial maderero en la zona de estudio Cap.3 Estudio del mercado.Definicion del producto.Mercado.Mercado interno para el proyecto.Analisis de la oferta.Mercado de exportacion.Estudio de precios.Sistema de comercializacion.Sistema de transporte para madera aserrada Cap.4 Tamano y localizacion de la planta.Introduccion.Tamano optimo.Localizacion de la planta Cap.5 Estudio de la materia prima.Lamadera como materia prima.Estudio tecnologico de dos especies.Analisis de datos y resultados.Criterios de clasificacion y aplicaciones industriales, segun las propiedades de la madera Cap.6 Ingenieria del proyecto planta de aserrado de la madera.Introduccion.Normalizacion del producto.Caracteristicas de las principales especies.Descripcion del proceso.Caracteristicas de la maquinaria y equipo para el proyecto.Plan de corta y programa de produccion Cap.7 Ingenieria del proyecto tratamiento del producto secado y preservacion de la madera.Introduccion.Secado de la madera.Sistema de secado.Contenido de humedad de la madera segun sus aplicaciones y usos.Secado artificial o en horno de secado.Diseno del horno de secado.Durabilidad y agentes destructores de la madera.Metodos de preservacion de la madera.Proceso de tratamiento y preservantes a utilizarse Cap.8 Sistema de aprovechamiento forestal.Introduccion.Temporada de trabajo.Horas efectivas de trabajo.Requerimiento de materia prima.Costos de apeo y troceo.Costo de arrastre o madereo.Costo de carguio.Sistema de transporte de materia prima.Formulacion de modelos matematicos para el aprovechamiento forestal Cap.9 Inversiones al proyecto.Inversiones en activos fijos.Inversiones en activos diferidos.Capital de operaciones.Plan de inversiones Cap.10 Costos gastos e ingresos del proyecto evaluacion economica y financiera.Costos de aprovechamiento forestal.Costos de produccion.Costos de administracion y servicios.Costo de comercializacion.Costo financiero.Ingresos al proyecto.Impuestos.Utilidades del proyecto.Fuentes y usos del proyecto.Evaluacion economica y financiera.Punto de equilibrio.