Sortilin knock-down alters the expression and distribution of cathepsin D and prosaposin and up-regulates the cation-dependent mannose-6-phosphate receptor in rat epididymal cells.

The selective transport to lysosomes can be mediated by either mannose-6-phosphate receptors (CD-MPR and CI-MPR) or sortilin. In mammalian epididymis, some lysosomal proteins are secreted into the lumen through unknown mechanisms. To investigate the underlying mechanisms of lysosomal protein transport in epididymal cells we studied the expression and distribution of cathepsin D (CatD) and prosaposin (PSAP) in a sortilin knocked down RCE-1 epididymal cell line (RCE-1 KD) in comparison with non-transfected RCE-1 cells. In RCE-1 cells, CatD was found in the perinuclear zone and co-localize with sortilin, whereas in RCE-1 KD cells, the expression, distribution and processing of the enzyme were altered. In turn, PSAP accumulated intracellularly upon sortilin knock-down and redistributed from LAMP-1-positive compartment to a perinuclear location, remaining co-localized with CatD. Interestingly, the sortilin knock-down induced CD-MPR overexpression and a redistribution of the receptor from the perinuclear zone to a dispersed cytoplasmic location, accompanied by an increased co-localization with CatD. The increase in CD-MPR could result from a compensatory response for the proper delivery of CatD to lysosomes in epididymal cells. The intracellular pathway taken by lysosomal proteins could be an approach for addressing further studies to understand the mechanism of exocytosis and therefore the role of these proteins in the epididymis.

First report of an egg-predator nemertean worm in crabs from the south-eastern Pacific coast: Carcinonemertes camanchaco sp. nov.

Nemertean worms belonging to the genus Carcinonemertes have been tied to the collapse of crab fisheries in the northeastern Pacific Ocean. A new species is described from egg masses of two commercial crabs, Cancer porteri and Romaleon setosum, inhabiting the central-north Chilean coast. This is the first species of Carcinonemertes described from the southeastern Pacific Ocean. Total body length of Carcinonemertes camanchaco sp. nov. ranged from 2.38 to 4.93 and from 4.29 to 8.92 mm, in males and females, respectively. Among others, traits that distinguish this new species from other previously described congeneric species include: presence of two gonad rows on each side of the intestine, a simple (not decorated) mucus sheath, and a relatively wide stylet basis. Maximum likelihood and Bayesian inference phylogenetic analyses distinguished this new species from all other species of Carcinonemertes with available cox1 sequences in GenBank. Prevalence and mean (± SD) intensity of C. camanchaco sp. nov. was 24% and 2.6 (± 2.07) worms per egg mass in C. porteri and 38.1% and 3.8 (± 2.4) worms per egg mass in R. setosum. The formal description of this new species represents the first step towards the understanding of this worm's impact on the health of crab fisheries in the southeastern Pacific Ocean.

Genetic homogeneity coupled with morphometric variability suggests high phenotypic plasticity in the sea louse Caligus rogercresseyi (Boxshall and Bravo, 2000), infecting farmed salmon (Salmo salar) along a wide latitudinal range in southern Chile.

The sea louse Caligus rogercresseyi is the most important pathogen causing "caligidosis" in the Chilean salmon industry. In this study, using cox1 gene, we evaluate the genetic variation of C. rogercresseyi from farmed Salmo salar along a latitudinal range (40°-52°S) in south Chile to determine whether morphological differences are explained by genetic or environmental factors. Female parasites were randomly collected from S. salar at five farms. Body variation was examined using multivariate analyses and genetic heterogeneity was explored with AMOVA. C. rogercresseyi exhibited significant morphometric variability among sites and parasites collected from >54°S were the longest ones. Parasites did not show genetic structure among farms. Thus, C. rogercresseyi infesting salmons is panmictic along an extensive latitudinal range in south Chile. The same genetic pattern can be explained by the frequent movement of parasitized S. salar among farms in that region. Phenotypic plasticity in parasites could be explained by natural or aquaculture-mediated environment variability. C. rogercreseyi from 54°S could favor the local spread of this disease, suggesting an immediate health risk for the recent salmon industry in that region. Further research is required to confirm genetic homogeneity of this parasite along its geographical distribution using more powerful markers (e.g. SNPs).

Rab Proteins: Insights into Intracellular Trafficking in Endometrium.

Rab proteins belong to the Ras superfamily of small monomeric GTPases. These G proteins are the main controllers of vesicular transport in every tissue, among them, the endometrium. They are in charge of to the functional subcellular compartmentalization and cargo transport between organelles and the plasma membrane. In turn, intracellular trafficking contributes to endometrial changes during the menstrual cycle, secretion to the uterine fluid, and trophoblast implantation; however, few reports analyze the role of Rab proteins in the uterus. In general, Rab proteins control the release of cytokines, growth factors, enzymes, hormones, cell adhesion molecules, and mucus. Further, the secretion of multiple compounds into the uterine cavity is required for successful implantation. Therefore, alterations in Rab-controlled intracellular transport likely impair secretory processes to the uterine fluid that may correlate with abnormal endometrial development and failed reproductive outcomes. Overall, they could explain recurrent miscarriages, female infertility, and/or assisted reproductive failure. Interestingly, estrogen (E2) and progesterone (P) regulate gene expression of Rab proteins involved in secretory pathways. This review aims to gather information regarding the role of Rab proteins and intracellular trafficking in the endometrium during the different menstrual phases, and in the generation of a receptive stage for embryo implantation, modulated by E2 and P. This knowledge might be useful for the development of novel reproductive therapies that overcome low implantation rates of assisted reproductive procedures.

Identificación y sensibilidad antimicrobiana de bacterias potencialmente responsables de infecciones nosocomiales en medicina veterinaria / Antimicrobial Identification and Sensitiveness of Bacteria Potentially Responsible for Nosocomial Infections in Veterinary Medicine

Resumen Las infecciones nosocomiales (IN) son definidas como aquellas infecciones que se presentan 48 a 72 horas después del ingreso a un hospital en Chile, causadas por bacterias con altos niveles de resistencia antibiótica. En Chile existen muy pocos estudios en medicina veterinaria, y se utilizan muchas veces los parámetros humanos. Por esto, la investigación tuvo como objetivo identificar y evaluar la sensibilidad antibiótica de la microbiota bacteriana, potencialmente causal de IN. Se realizó un estudio de corte transversal a 20 objetos en una clínica veterinaria durante el mes de junio del año 2012. Se evidenció desarrollo bacteriano en el 85 % de los elementos muestreados. Se logró aislar cepas de Staphylococcus epidermidis (50 %), Acinetobacter spp. (15 %), Enterococcus faecalis (15 %), Escherichia coli (10 %), Proteus mirabilis (5 %) y Bacillus subtilis (5 %). Se logró identificar especies bacterianas, con potencial para generar infecciones nosocomiales, con un grado medio de resistencia antimicrobiana.

Abstract Nosocomial infections (NI) are defined as those occurring within 48-72 hours after being admitted in a hospital, and caused by bacteria with high antibiotic resistance levels. In Chile there are very few studies in veterinary medicine on this issue and most of the times human parameters are used instead. Therefore, this research aimed to identify and evaluate the antibiotic sensitiveness of the bacterial microbiota potentially causing NI. A cross-sectional study was conducted using 20 objects from a veterinary clinic during June 2012. Bacterial growth was observed in 85% of the sampled objects. The isolated strains include Staphylococcus epidermidis (50%), Acinetobacter spp. (15%), Enterococcus faecalis (15%), Escherichia coli (10%), Proteus mirabilis (5%), and Bacillus subtilis (5%). Bacteria species were identified with a potential to cause nosocomial infections of medium-grade antimicrobial resistance.

Evaluación microbiológica de dispositivos móviles en personal quirúrgico de una institución de salud, Pereira, Colombia, 2018 / Microbiological analysis of mobile devices of surgical personnel working at a health care institution in Pereira, Colombia, 2018

Introducción: El uso del teléfono celular se ha vuelto común en áreas del hospital, incluida la sala de operaciones, aumentando el riesgo de contaminación cruzada y las altas tasas de infecciones del sitio quirúrgico. Objetivo: Determinar la contaminación microbiana en dispositivos móviles del personal quirúrgico de una Institución de Salud de Pereira, Colombia en el primer semestre del año 2018. Materiales y métodos: Estudio descriptivo observacional, transversal. Se tomaron 10 dispositivos móviles del personal del área de quirófano al azar y a cada uno se le realizó frotis con hisopo estéril al estuche o cobertura del celular. Las muestras se transportaron en tubos de ensayo con agua peptonada al 1% hasta el momento de la siembra; las muestras se analizaron por técnica microbiológica recuento en placa profunda. Resultados: En las muestras se encontró un promedio de 93 UFC (Unidades Formadoras de Colonias) de mesófilos aerobios, 13 UFC de coliformes totales, 22 UFC de mohos y levaduras. Conclusión: El análisis microbiológico permitió detectar la presencia de elevadas cantidades de unidades formadoras de colonias que podrían contribuir de manera significativa a incrementar las tasas en infecciones asociadas a la atención en salud.

Introduction: Cell phone use in hospital areas such as surgery rooms has become a common practice, which has increased both the risk of cross-contamination and the rates of surgical site infections. Objective: To determine microbial contamination of mobile devices belonging to surgical staff of a Health Care Institution in Pereira, Colombia during the first semester of 2018. Materials and methods: An observational, cross-sectional, descriptive study was conducted with a sample of ten mobile devices randomly chosen from health personnel working in the surgical room. Surface samples were collected from cell phone cases and bags using sterile swabs that were were kept in a 1% peptone salt solution until a deep plate count assay was performed on them. Results: The plate count confirmed the presence of mesophilic aerobes (93 CFUs), total coliforms (13 CFUs) as well as yeasts and molds (22 CFUs). Conclusion: The microbiological analysis showed elevated numbers of colony-forming units that could significantly increase the rates of infections associated with health care settings.

Temperature and pCO2 jointly affect the emergence and survival of cercariae from a snail host: implications for future parasitic infections in the Humboldt Current system.

Ocean warming and acidification are general consequences of rising atmospheric CO2 concentrations. In addition to future predictions, highly productive systems such as the Humboldt Current System are characterized by important variations in both temperature and pCO2 level, but how these physical-chemical ocean changes might influence the transmission and survival of parasites has not been assessed. This study experimentally evaluated the effects of temperature (14, 18 and 25 °C) and the combined effects of temperature (∼15 and 20 °C) and pCO2 level (∼500 and 1400 microatmospheres (µatm) on the emergence and survival of two species of marine trematodes-Echinostomatidae gen. sp. and Philophthalmidae gen. sp.-both of which infect the intertidal snail Echinolittorina peruviana. Snails were collected from intertidal rocky pools in a year-round upwelling area of the northern Humboldt Current System (23°S). Two experiments assessed parasite emergence and survival by simulating emersion-immersion tidal cycles. To assess parasite survival, 2 h old cercariae (on average) were taken from a pool of infected snails incubated at 20-25 °C, and their mortality was recorded every 6 h until all the cercariae were dead. For both species, a trade-off between high emergence and low survival of cercariae was observed in the high temperature treatment. Species-specific responses to the combination of temperature and pCO2 levels were also observed: the emergence of Echinostomatidae cercariae was highest at 20 °C regardless of the pCO2 levels. By contrast, the emergence of Philophthalmidae cercariae was highest at elevated pCO2 (15 and 20 °C), suggesting that CO2 may react synergistically with temperature, increasing transmission success of this parasite in coastal ecosystems of the Humboldt Current System where water temperature and pH are expected to decrease. In conclusion, our results suggest that integrating temperature-pCO2 interactions in parasite studies is essential for understanding the consequence of climate change in future marine ecosystem health.

Determining Intermediate Hosts for Opecoelidae and Microphallidae Species (Platyhelminthes: Trematoda) in the Southeastern Pacific Coast, Using Molecular Markers.

Metacercarial stages of digeneans were collected from decapod crustaceans inhabiting intertidal rocky zones in central Chile. The digeneans were identified through a molecular analysis based on the V4 region of the 18S ribosomal RNA gene. We analyzed 356 crustaceans belonging to 10 species, 115 intertidal fish belonging to 6 species, and 4 specimens of 1 coastal bird species. In total, 74.1% of crustaceans were parasitized with metacercariae. We found 1 species of Opecoelidae. This species showed low genetic divergence (0% and 0.1%) with adult digeneans found in intertidal fish and with the species Helicometrina labrisomi infesting a subtidal fish from northern Chile (Labrisomus philippii). Additionally, we found 2 species of Microphallidae, 1 closely related to Maritrema (1.3% genetic distance) and the other related to Microphallus (5% genetic distance). Therefore, our findings showed that the decapod crustaceans are relevant hosts in food webs from the southeastern Pacific coast. Furthermore, we found 5 species of crustaceans as second intermediate hosts for H. labrisomi and 2 species as secondary intermediate hosts for 2 Microphallidae, which contribute to elucidate parts of their life cycles through molecular markers and extended the host distribution of H. labrisomi in the southeastern Pacific coast.

Características antropométricas de futbolistas profesionales chilenos / Anthropometric characteristics of chilean professional football players

La tendencia en la formación en el futbol, determina que las variables antropométricas sean importantes al momento de definir entre otras cosas, la posición de juego más idónea. Estos parámetros morfológicos son una parte esencial de la evaluación y selección de los deportistas. Estos patrones permiten determinar la evolución física del futbolista desde temprana edad, lo que ayuda enormemente en la intervención de los mismos. Se evaluó antropométricamente, a un total de 406 futbolistas profesionales, pertenecientes a 15 clubes chilenos, de los cuales 12 pertenecen a Primera División y 3 clubes a Primera B. Existen diferencias en algunas variables antropométricas básicas por posición de juego, que nos ponen en desventaja frente a otros jugadores internacionales, como la estatura. No hay diferencias en el Phantom ni en el somatotipo por posición de juego, es decir tienen variables relativamente homogéneas. Esto podría hacernos pensar que un jugador puede desempeñarse en cualquier posición de juego, pero hemos visto que la estatura, el peso y la masa grasa son distintas por cada posición, por lo que cada jugador tiene su función y sus propias características en las que los entrenadores y preparadores deben fijarse al momento de ubicar a cada jugador en su función específica.

The trend in the formation in football, determines that the anthropometric variables are important when defining among other things, the most suitable playing position. These morphological parameters are an essential part of the evaluation and selection of athletes. These patterns can determine the physical evolution of football from an early age, which helps tremendously in the operation thereof. Anthropometrically, a total of 406 professional footballers from 15 Chilean clubs were evaluated, of which 12 belong to the first division clubs and 3 to First B. There are differences in some basic anthropometric variables by playing position, which places us at a disadvantage compared to other international players, such as height. No differences in the Phantom or the somatotype by playing position, i.e. variables are relatively homogeneous. This could suggest that a player can perform at any position in the game, but we have seen that the height, weight and fat mass are different for each position, so that each player has their own characteristics and their role in the coaches and trainers should be fixed at the time of placing each player on their specific function.

Rab11-family of interacting protein 2 associates with chlamydial inclusions through its Rab-binding domain and promotes bacterial multiplication.

Chlamydia trachomatis, an obligate intracellular pathogen, survives within host cells in a special compartment named 'inclusion' and takes advantage of host vesicular transport pathways for its growth and replication. Rab GTPases are key regulatory proteins of intracellular trafficking. Several Rabs, among them Rab11 and Rab14, are implicated in chlamydial development. FIP2, a member of the Rab11-Family of Interacting Proteins, presents at the C-terminus a Rab-binding domain that interacts with both Rab11 and Rab14. In this study, we determined and characterized the recruitment of endogenous and GFP-tagged FIP2 to the chlamydial inclusions. The recruitment of FIP2 is specific since other members of the Rab11-Family of Interacting Proteins do not associate with the chlamydial inclusions. The Rab-binding domain of FIP2 is essential for its association. Our results indicate that FIP2 binds to Rab11 at the chlamydial inclusion membrane through its Rab-binding domain. The presence of FIP2 at the chlamydial inclusion favours the recruitment of Rab14. Furthermore, our results show that FIP2 promotes inclusion development and bacterial replication. In agreement, the silencing of FIP2 decreases the bacterial progeny. C. trachomatis likely recruits FIP2 to hijack host intracellular trafficking to redirect vesicles full of nutrients towards the inclusion.

Golgi-associated Rab14, a new regulator for Chlamydia trachomatis infection outcome.

Chlamydia trachomatis is the causing agent of the most frequent bacterial sexually-transmitted diseases worldwide and is an underlying cause of chronic pelvic inflammatory diseases and cervical cancer. It is an obligate intracellular bacterium that establishes a close relationship with the Golgi complex and parasites the biosynthetic machinery of host cells. In a recent study, we have demonstrated that Rab14, a newly-described Golgi-associated Rab, is involved in the delivery of sphingolipids to the growing bacteria-containing vacuole. The interference with Rab14-controlled trafficking pathways delays chlamydial inclusion enlargement, decreases bacterial lipid uptake, negatively impact on bacterial differentiation, and reduces bacterial progeny and infectivity. C. trachomatis manipulation of host trafficking pathways for the acquisition of endogenously-biosynthesized nutrients arises as one of the characteristics of this highly evolved pathogen. The development of therapeutic strategies targeted to interfere with bacterium-host cell interaction is a new challenge for pharmacological approaches to control chlamydial infections.

Reconstitution of recycling from the phagosomal compartment in streptolysin O-permeabilized macrophages: role of Rab11.

By phagocytosis, macrophages engulf large particles, microorganisms and senescent cells in vesicles called phagosomes. Many internalized proteins rapidly shuttle back to the plasma membrane following phagosome biogenesis. Here, we report a new approach to the study of recycling from the phagosomal compartment: streptolysin O- (SLO) permeabilized macrophages. In this semi-intact cell system, energy and cytosol are required to efficiently reconstitute recycling transport. Addition of GDPbetaS strongly inhibits this transport step, suggesting that a GTP-binding protein modulates the dynamics of cargo exit from the phagosomal compartment. GTPases of the Rab family control vesicular trafficking, and Rab11 is involved in transferrin receptor recycling. To unravel the role of Rab11 in the phagocytic pathway, we added recombinant proteins to SLO-permeabilized macrophages. Rab11:S25N, a negative mutant, strongly diminishes the release of recycled proteins from phagosomes. In contrast, wild type Rab11 and its positive mutant (Rab11:Q70L) favor this vesicular transport event. Using biochemical and morphological assays, we confirm that overexpression of Rab11:S25N substantially decreases recycling from phagosomes in intact cells. These findings show the requirement of a functional Rab11 for the retrieval to the plasma membrane of phagosomal content. SLO-permeabilized macrophages likely constitute a useful tool to identify new molecules involved in regulating transport along the phagocytic pathway.

Rab coupling protein associates with phagosomes and regulates recycling from the phagosomal compartment.

The Rab coupling protein (RCP) is a recently identified novel protein that belongs to the Rab11-FIP family. RCP interacts specifically with Rab4 and Rab11, small guanosine-5'-triphosphatases that function as regulators along the endosomal recycling pathway. We used fluorescence confocal microscopy and biochemical approaches to evaluate the participation of RCP during particle uptake and phagosome maturation. In macrophages, RCP is predominantly membrane-bound and displays a punctuate vesicular pattern throughout the cytoplasm. RCP is mainly associated with transferrin-containing structures and Rab11-labeled endosomes. Overexpression of H13, the carboxyl-terminal region of RCP that contains the Rab binding domain, results in an abnormal endosomal compartment. Interestingly, we found that RCP is associated as discrete patches or protein domains to early phagosomal membranes. In macrophages, overexpression of full-length RCP stimulates recycling from the phagosomal compartment, whereas overexpression of H13 diminishes this vesicular transport step. It is likely that acting as an intermediate between Rab4 and Rab11, RCP regulates membrane flux along the phagocytic pathway via recycling events.