SPIM-Flow: An Integrated Light Sheet and Microfluidics Platform for Hydrodynamic Studies of Hydra.

Selective plane illumination microscopy (SPIM), or light sheet microscopy, is a powerful imaging approach. However, access to and interfacing microscopes with microfluidics have remained challenging. Complex interfacing with microfluidics has limited the SPIM's utility for studying the hydrodynamics of freely moving multicellular organisms. We developed SPIM-Flow, an inexpensive light sheet platform that enables easy integration with microfluidics. We used SPIM-Flow to investigate the hydrodynamics of a freely moving Hydra polyp via particle tracking in millimeter-sized chambers. Initial experiments across multiple animals, feeding on a chip (Artemia franciscana nauplii used as food), and baseline behaviors (tentacle swaying, elongation, and bending) indicated the organisms' health inside the system. Fluidics were used to investigate Hydra's response to flow. The results suggested that the animals responded to an established flow by bending and swaying their tentacles in the flow direction. Finally, using SPIM-Flow in a proof-of-concept experiment, the shear stress required to detach an animal from a surface was demonstrated. Our results demonstrated SPIM-Flow's utility for investigating the hydrodynamics of freely moving animals.

IL-10-Dependent Amelioration of Chronic Inflammatory Disease by Microdose Subcutaneous Delivery of a Prototypic Immunoregulatory Small Molecule.

One of the interventional strategies to reestablish the immune effector/regulatory balance, that is typically altered in chronic inflammatory diseases (CID), is the reinforcement of endogenous immunomodulatory pathways as the one triggered by interleukin (IL)-10. In a recent work, we demonstrated that the subcutaneous (sc) administration of an IL-10/Treg-inducing small molecule-based formulation, using a repetitive microdose (REMID) treatment strategy to preferentially direct the effects to the regional immune system, delays the progression of atherosclerosis. Here we investigated whether the same approach using other IL-10-inducing small molecule, such as the safe, inexpensive, and widely available polyphenol curcumin, could induce a similar protective effect in two different CID models. We found that, in apolipoprotein E deficient mice, sc treatment with curcumin following the REMID strategy induced atheroprotection that was not consequence of its direct systemic lipid-modifying or antioxidant activity, but instead paralleled immunomodulatory effects, such as reduced proatherogenic IFNγ/TNFα-producing cells and increased atheroprotective FOXP3+ Tregs and IL-10-producing dendritic and B cells. Remarkably, when a similar strategy was used in the neuroinflammatory model of experimental autoimmune encephalomyelitis (EAE), significant clinical and histopathological protective effects were evidenced, and these were related to an improved effector/regulatory cytokine balance in restimulated splenocytes. The essential role of curcumin-induced IL-10 for neuroprotection was confirmed by the complete abrogation of the clinical effects in IL-10-deficient mice. Finally, the translational therapeutic prospection of this strategy was evidenced by the neuroprotection observed in mice starting the treatment one week after disease triggering. Collectively, results demonstrate the power of a simple natural IL-10-inducing small molecule to tackle chronic inflammation, when its classical systemic and direct pharmacological view is shifted towards the targeting of regional immune cells, in order to rationally harness its immunopharmacological potential. This shift implies that many well-known IL-10-inducing small molecules could be easily reformulated and repurposed to develop safe, innovative, and accessible immune-based interventions for CID.

Desempeño de Capitán de la Sabana (Eremophilus mutisii) en estadios tempranos de desarrollo / Performance of the Captain of the Savannah, (Eremophilus mutisii) in early stages of development

RESUMEN Son varios los esfuerzos realizados en el pez capitán de la sabana Eremophilus mutisii para su uso y conservación; sin embargo, a pesar de los avances logrados, no se ha conseguido superar las dificultades de la larvicultura en esta especie, razón por la cual, se realizó la primera descripción de las etapas iniciales de vida, con el fin de contribuir y de mejorar los inicios de la larvicultura. Las larvas utilizadas fueron obtenidas de reproductores maduros de capitán de la sabana. Posteriormente, se procedió a describir el volumen y determinación de absorción del saco vitelino, la abertura máxima de la bucal, primera alimentación, preferencia alimentaria y la evaluación de variables productivas, como ganancia de peso, tasa de crecimiento específica y sobrevivencia. El consumo del saco vitelino, se completó en la 201 Hora Post Eclosión (HPE), equivalente a más del 90% de absorción, la apertura máxima bucal es alcanzada a las 126 HPE. Esta especie presenta un alto de grado de aceptación a la alimentación con quistes de artemia, a partir del 11 Día Post Eclosión(DPE) y, posteriormente, a alimento artificial, desde los 47 DPE; lo anterior, se convierte en un aporte para la obtención e identificación de mecanismos y estrategias que promuevan el uso y la conservación en cautiverio de E. mutisii, con el fin de fomentar el aprovechamiento acuícola en este especie, que se encuentra en amenaza de extinción y puede ser una fuente alimenticia, que contribuya a la seguridad alimentaria de la población Cundiboyacense.

ABSTRACT There are several efforts made in the savannah captain fish Eremophilus mutisii for its use and conservation; however, despite the progress made, it has not been possible to overcome the difficulties of larviculture in this species, reason for to make the first description of the initial stages of life for contribute and improve the beginnings of larviculture. The larvae used were obtained from mature savanna captain breeders. Subsequently, the volume and determination of the absorption of the yolk sac, the maximum opening of the mouth, first feeding, food preference and the evaluation of productive variables such as: weight gain, specific growth rate and survival were described. The consumption of the yolk sac was completed at 201 Hour Post Hatching (HPE), equivalent to more than 90% absorption, the maximum mouth opening is reached at 126 HPE. This species has a high degree of acceptance when feeding with artemia cysts from 11 Day post-hatching (DPE) and later to artificial food from 47 DPE, This to becomes a contribution to the obtaining and identification of mechanisms and strategies that promote the use and conservation of E. mutisii in captivity, in order to promote the use of aquaculture in this species that is threatened with extinction and can be a food source that contributes to the food security of the Cundiboyacense population.

Green Coffee Extract Improves Cardiometabolic Parameters and Modulates Gut Microbiota in High-Fat-Diet-Fed ApoE-/- Mice.

Chlorogenic acids (CGA) are the most abundant phenolic compounds in green coffee beans and in the human diet and have been suggested to mitigate several cardiometabolic risk factors. Here, we aimed to evaluate the effect of a water-based standardized green coffee extract (GCE) on cardiometabolic parameters in ApoE-/- mice and to explore the potential underlying mechanisms. Mice were fed an atherogenic diet without (vehicle) or with GCE by gavage (equivalent to 220 mg/kg of CGA) for 14 weeks. We assessed several metabolic, pathological, and inflammatory parameters and inferred gut microbiota composition, diversity, and functional potential. Although GCE did not reduce atherosclerotic lesion progression or plasma lipid levels, it induced important favorable changes. Specifically, improved metabolic parameters, including fasting glucose, insulin resistance, serum leptin, urinary catecholamines, and liver triglycerides, were observed. These changes were accompanied by reduced weight gain, decreased adiposity, lower inflammatory infiltrate in adipose tissue, and protection against liver damage. Interestingly, GCE also modulated hepatic IL-6 and total serum IgM and induced shifts in gut microbiota. Altogether, our results reveal the cooccurrence of these beneficial cardiometabolic effects in response to GCE in the same experimental model and suggest potential mediators and pathways involved.

ESTUDIO HISTOLÓGICO Y MORFOLÓGICO DEL DESARROLLO EMBRIONARIO DEL PEZ CAPITÁN DE LA SABANA (Eremophilus mutisii) / HISTOLOGICAL AND MORPHOLOGICAL STUDY OF THE EMBRIONARY DEVELOPMENT OF THE CAPITAN DE LA SABANA (Eremophilus mutisii)

RESUMEN Eremophilus mutisii, pez catalogado en estado como vulnerable, que por su importancia sociocultural y ecológica genera retos investigativos, uno de ellos, conocer sus características histológicas del desarrollo embrionario, desde la fecundación hasta la eclosión. Para ello, se obtuvieron embriones de individuos adultos y maduros sexualmente, capturados en la represa del Sisga, ubicada en el municipio de Chocontá, Cundinamarca, inducidos sexualmente con extracto pituitario de carpa. Los ovocitos y el semen fueron obtenidos por presión abdominal 24 horas después de la última dosis, a 14±1°C. La toma de muestras, se realizó cada 60 minutos, durante las primeras doce horas post-fertilización (HPF); a partir de aquí y hasta la eclosión, las muestras fueron fijadas en formol Bufferado al 4%, cada 6 horas y sometidas a rutina de histología. Los resultados mostraron que los óvulos fertilizados presentaron forma esférica, color verde claro y diámetro promedio de 1270µm. El primer clivaje, se observó a las 2HPF; la blastulación inició a las 9HPF; los movimientos epibólicos y la gastrulación, se observaron desde la 12HPF hasta la 30HPF, con el cierre del blastoporo. A las 60HPF, se evidenció el desprendimiento de la cola del saco vitelino y a las 66HPF, se dio inicio a los primeros movimientos autónomos. Finalmente, a las 72HPF inició la eclosión, mostrando las larvas ausencia de pigmentación y evidenciándose un tracto digestivo rudimentario, sin abertura bucal y anal. Se observó la presencia de abundante saco vitelino, con movimientos verticales continuos de la larva.

ABSTRACT Eremophilus mutisii, due to its vulnerable condition and its potential for fish farming, generates research challenges; one of them is to know the embryonic changes of the E. mutisii based on morphological and histological features from fertilization to hatching. The embryos were obtained from adult and sexually mature individuals of the Capitán de la Sabana (Eremophilus mutisii), sexually induced with carp pituitary extract. Oocytes and semen were obtained by abdominal pressure 24 hours after the last dose at 14±1°C. Sampling was performed every 60 minutes for the first twelve hours´ post-fertilization (HPF); from here on at hatching, the samples were fixed every 6 hours. The embryos were fixed in 4% formalin for observing the morphological features and for the histological studies. Results revealed that the fertilized eggs had spherical shape, light green color and average diameter of 1270µm. The first cleavage at 2 HPF was observed, the blastulation began at 9 HPF. Epibolic movements and gastrulation were observed from 12 HPF to 30 HPF with blastopore closure. At 60 HPF the yolk sac tail detachment was evident and at 66 HPF the first autonomous movements began. Finally, at 72 HPF, the hatching began, showing absence of pigmentation and evidencing a rudimentary digestive tract without oral and anal opening. It was observed the presence of abundant yolk sac with continuous vertical movements of the larva.

DHEA increases epithelial markers and decreases mesenchymal proteins in breast cancer cells and reduces xenograft growth.

Breast cancer is one of the most common neoplasias and the leading cause of cancer death in women worldwide. Its high mortality rate is linked to a great metastatic capacity associated with the epithelial-mesenchymal transition (EMT). During this process, a decrease in epithelial proteins expression and an increase of mesenchymal proteins are observed. On the other hand, it has been shown that dehydroepiandrosterone (DHEA), the most abundant steroid in human plasma, inhibits migration of breast cancer cells; however, the underlying mechanisms have not been elucidated. In this study, the in vitro effect of DHEA on the expression pattern of some EMT-related proteins, such as E-cadherin (epithelial), N-cadherin, vimentin and Snail (mesenchymal) was measured by Western blot and immunofluorescence in MDA-MB-231 breast cancer cells with invasive, metastatic and mesenchymal phenotype. Also, the in vivo effect of DHEA on xenograft tumor growth in nude mice (nu-/nu-) and on expression of the same epithelial and mesenchymal proteins in generated tumors was evaluated. We found that DHEA increased expression of E-cadherin and decreased N-cadherin, vimentin and Snail expression both in MD-MB-231 cells and in the formed tumors, possibly by DHEA-induced reversion of mesenchymal phenotype. These results were correlated with a tumor size reduction in mouse xenografts following DHEA administration either a week earlier or concurrent with breast cancer cells inoculation. In conclusion, DHEA could be useful in the treatment of breast cancer with mesenchymal phenotype.

Secretome derived from breast tumor cell lines alters the morphology of human umbilical vein endothelial cells.

Metastases, responsible for most of the solid tumor associated deaths, require angiogenesis and changes in endothelial cells. In this work, the effect of the secretomes of three breast tumor cell lines (MCF-7, MDA-MB-231 and ZR-75-30) on human umbilical vein endothelial cells (HUVEC) morphology was investigated. HUVEC treated with secretomes from breast cells were analyzed by confocal and time-lapse microscopy. Secretomes from ZR-75-30 and MDA-MB-231 cells modify the morphology and adhesion of HUVEC. These changes may provoke the loss of endothelial monolayer integrity. In consequence, tumor cells could have an increased access to circulation, which would then enhance metastasis.

Internalization of titanium dioxide nanoparticles by glial cells is given at short times and is mainly mediated by actin reorganization-dependent endocytosis.

Many nanoparticles (NPs) have toxic effects on multiple cell lines. This toxicity is assumed to be related to their accumulation within cells. However, the process of internalization of NPs has not yet been fully characterized. In this study, the cellular uptake, accumulation, and localization of titanium dioxide nanoparticles (TiO2 NPs) in rat (C6) and human (U373) glial cells were analyzed using time-lapse microscopy (TLM) and transmission electron microscopy (TEM). Cytochalasin D (Cyt-D) was used to evaluate whether the internalization process depends of actin reorganization. To determine whether the NP uptake is mediated by phagocytosis or macropinocytosis, nitroblue tetrazolium (NBT) reduction was measured and the 5-(N-ethyl-N-isopropyl)-amiloride was used. Expression of proteins involved with endocytosis and exocytosis such as caveolin-1 (Cav-1) and cysteine string proteins (CSPs) was also determined using flow cytometry. TiO2 NPs were taken up by both cell types, were bound to cellular membranes and were internalized at very short times after exposure (C6, 30 min; U373, 2h). During the uptake process, the formation of pseudopodia and intracellular vesicles was observed, indicating that this process was mediated by endocytosis. No specific localization of TiO2 NPs into particular organelles was found: in contrast, they were primarily localized into large vesicles in the cytoplasm. Internalization of TiO2 NPs was strongly inhibited by Cyt-D in both cells and by amiloride in U373 cells; besides, the observed endocytosis was not associated with NBT reduction in either cell type, indicating that macropinocytosis is the main process of internalization in U373 cells. In addition, increases in the expression of Cav-1 protein and CSPs were observed. In conclusion, glial cells are able to internalize TiO2 NPs by a constitutive endocytic mechanism which may be associated with their strong cytotoxic effect in these cells; therefore, TiO2 NPs internalization and their accumulation in brain cells could be dangerous to human health.

Phenotype of the anti-Rickettsia CD8(+) T cell response suggests cellular correlates of protection for the assessment of novel antigens.

The obligately intracellular bacteria Rickettsia infect endothelial cells and cause systemic febrile diseases that are potentially lethal. No vaccines are currently available and current knowledge of the effective immune response is limited. Natural and experimental rickettsial infections provide strong and cross-protective cellular immunity if the infected individual survives the acute infection. Although resistance to rickettsial infections is attributed to the induction of antigen-specific T cells, particularly CD8(+) T cells, the identification and validation of correlates of protective cellular immunity against rickettsial infections, an important step toward vaccine validation, remains a gap in this field. Here, we show that after a primary challenge with Rickettsia typhi in the C3H mouse model, the peak of anti-Rickettsia CD8(+) T cell-mediated responses occurs 7 days post-infection (dpi), which coincides with the beginning of rickettsial clearance. At this time point, both effector-type and memory-type CD8(+) T cells are present, suggesting that 7 dpi is a valid time point for the assessment of CD8(+) T cell responses of mice previously immunized with protective antigens. Based on our results, we suggest four correlates of cellular protection for the assessment of protective rickettsial antigens: (1) production of IFN-γ by antigen-experienced CD3(+)CD8(+)CD44(high) cells, (2) production of Granzyme B by CD27(low)CD43(low) antigen-experienced CD8(+) T cells, (3) generation of memory-type CD8(+) T cells [Memory Precursor Effector Cells (MPECs), as well as CD127(high)CD43(low), and CD27(high)CD43(low) CD8(+) T cells], and (4) generation of effector-like memory CD8(+) T cells (CD27(low)CD43(low)). We propose that these correlates could be useful for the general assessment of the quality of the CD8(+) T cell immune response induced by novel antigens with potential use in a vaccine against Rickettsia.

Discovery of novel cross-protective Rickettsia prowazekii T-cell antigens using a combined reverse vaccinology and in vivo screening approach.

Rickettsial agents are some of the most lethal pathogens known to man. Among them, Rickettsia prowazekii is a select agent with potential use for bioterrorism; yet, there is no anti-Rickettsia vaccine commercially available. Owing to the obligate intracellular lifestyle of rickettsiae, CD8(+) T cells are indispensable for protective cellular immunity. Furthermore, T cells can mediate cross-protective immunity between different pathogenic Rickettsia, a finding consistent with the remarkable similarity among rickettsial genomes. However, Rickettsia T cell antigens remain unidentified. In the present study, we report an algorithm that allowed us to identify and validate four novel R. prowazekii vaccine antigen candidates recognized by CD8(+) T cells from a set of twelve in silico-defined protein targets. Our results highlight the importance of combining proteasome-processing as well as MHC class-I-binding predictions. The novel rickettsial vaccine candidate antigens, RP778, RP739, RP598, and RP403, protected mice against a lethal challenge with Rickettsia typhi, which is indicative of cross-protective immunity within the typhus group rickettsiae. Together, our findings validate a reverse vaccinology approach as a viable strategy to identify protective rickettsial antigens and highlight the feasibility of a subunit vaccine that triggers T-cell-mediated cross-protection among diverse rickettsiae.

Paneth cell identification in the small intestine of guinea pig offsprings (Cavia porcellus).

The aim of this study was to determine the presence, number, and morphometrical characteristics of Paneth cells (PC) in the small intestine of guinea pigs during lactation. We used 48 pups from 0 to 15 days old. Samples from small intestine were fixed in 10% buffered formaldehyde (pH 7.4) and processed for histological and morphometrical studies using hematoxylin and eosin (HE), Phloxine tartrazine or Masson's Trichome staining, or immunohistochemistry for lysozyme. PC were morphologically identified at day 2 using Masson's Trichome or Phloxine tartrazine stainings, and at day 4 using HE, whereas using immunohistochemistry they were recognized from birth. Morphometrical differences were found between the intestinal sections at each age studied, and within each section during the first weeks of life. In all developmental stage, the highest number of PC was observed in the duodenum of 13 days old guinea pigs. Our results confirm the presence of PC in the small intestine of guinea pigs from birth.

Discovery of a protective Rickettsia prowazekii antigen recognized by CD8+ T cells, RP884, using an in vivo screening platform.

Rickettsia prowazekii has been tested for biological warfare due to the high mortality that it produces after aerosol transmission of very low numbers of rickettsiae. Epidemic typhus, the infection caused by these obligately intracellular bacteria, continues to be a threat because it is difficult to diagnose due to initial non-specific symptoms and the lack of commercial diagnostic tests that are sensitive and specific during the initial clinical presentation. A vaccine to prevent epidemic typhus would constitute an effective deterrent to the weaponization of R. prowazekii; however, an effective and safe vaccine is not currently available. Due to the cytoplasmic niche of Rickettsia, CD8(+) T-cells are critical effectors of immunity; however, the identification of antigens recognized by these cells has not been systematically addressed. To help close this gap, we designed an antigen discovery strategy that uses cell-based vaccination with antigen presenting cells expressing microbe's proteins targeted to the MHC class I presentation pathway. We report the use of this method to discover a protective T-cell rickettsial antigen, RP884, among a test subset of rickettsial proteins.

Bullying, redes de apoyo social y funcionamiento familiar en adolescentes de una institución educativa de Santander, Colombia / Bullying, social support networks and family functioning in adolescents in an educational institution Santander, Colombia

La presente investigación tuvo como objetivo caracterizar el bullying y la relación entre redes de apoyo social y funcionamiento familiar percibido por los adolescentes de una institución educativa. Se propuso un diseño no experimental de tipo descriptivo-correlacional con el que se recogieron datos exclusivamente de Floridablanca, Santander. La población estuvo conformada por 304 estudiantes de los grados sextos, novenos y undécimos de una institución educativa pública; las edades oscilaron entre los 10 a 18 años. Como instrumentos se utilizaron tres cuestionarios: el Cuestionario Paredes Lega y Vernon, (2006) para detección del bullying, el APGAR Familiar para medir el funcionamiento familiar, y el Cuestionario MOS de apoyo social. El análisis de la información se trabajó con el programa estadístico SPSS 17.0. Los resultados evidencian que en la institución educativa se presentan conductas asociadas al fenómeno de bullying (agresión, exclusión, intimidación), el 30,5 % de todos los estudiantes admitió que alguna vez ha agredido de diferentes formas a un compañero o compañera ridiculizándolo, golpeándolo, excluyéndolo o por medio de la amenaza, siendo la más repetitiva la ridiculización con un 44,5 %, los resultados demostraron la presencia de bullying en el 22,8% de los estudiantes evaluados. Finalmente, se concluye en la investigación, la importancia de fortalecer las redes de apoyo dentro del contexto escolar, ya que no está cumpliendo la función de brindar apoyo hacia los jóvenes que presentan estos comportamientos, específicamente los que son agredidos, lo que demuestra la necesidad de hacer más partícipes a los docentes, familiares y pares, en las dinámicas que se gestan al interior de la institución educativa que apunten a la construcción de redes de apoyo sólidas en los estudiantes.

The present study aimed to characterize the relationship between bullying and social support networks and family functioning adolescents perceived by an educational institution. We proposed a nonexperimental descriptive-correlational data which was collected exclusively Floridablanca, Santander. The sample consisted of 304 students in grades sixth, ninth and eleventh in a public school, the ages ranged from 10 to 18. As instruments used three questionnaires: Questionnaire Paredes Lega and Vernon (2006) for detection of bullying, the Family APGAR to measure family functioning, and social support questionnaire MOS. The data analysis was worked with SPSS 17.0. The results show that in school behaviors are associated with the phenomenon of bullying (aggression, exclusion, bullying), 30.5% of all students admitted to have ever attacked in different ways to a partner ridiculing, beating , or by excluding it from the threat, the most repetitive ridicule with 44.5%, the results showed the presence of "bullying" in 22.8% of the students tested. Finally, we conclude in research, the importance of strengthening support networks within the school context, as it is not fulfilling the role of providing support to young people who have these behaviors, specifically those who are bullied, which demonstrates the need for more teachers involved, family and peers, the dynamics that are developing within the educational institution aimed at building strong support networks for students.

Paragonimus mexicanus en hospederos naturales en el Estado Sucre, Venezuela / Paragonimus mexicanus in natural hosts from two municipalities of the Sucre State, Venezuela

Se estudió la infección por Paragonimus mexicanus en el rabipelado (Didelphis marsupialis) y cangrejo de agua dulce (Eudaniela garmani) en los municipios Sucre y Montes del Estado Sucre, Venezuela. Utilizando en la captura de ambas especies, trampas Sherman y redes de mano. En Sucre la prevalencia de parásitos fue 13,33% con una intensidad de 1,50 parásitos/rabipelado, y en Montes la prevalencia fue 31,88% e intensidad de 6,82 parásitos/rabipelado. En 634 cangrejos de Montes, la prevalencia de metacercarias fue 17,98% y 4,07 en 123 cangrejos de Sucre. En ambos municipios no hubo diferencias significativas entre prevalencia, intensidad y el sexo de los cangrejos, pero si una correlación positiva y significativa con la talla. En Sucre la intensidad de infección por individuo fue mayor (8,20) que en Montes (5,31). Se concluye que la infección fue más intensa en cangrejos hembras y en los de mayor talla. Estos resultados indican la existencia de un foco zoonótico en ambas áreas y la necesidad de continuar las investigaciones para determinar los elementos relevantes de esta interrelación parasitaria.

The Paragonimus mexicanus infection was studied in the opposum Didelphis marsupialis and crab Eudaniela garmani from the Sucre and Montes municipalities of Sucre State, Venezuela. In Sucre, prevalence of adult parasites in opposums was 13. 33% with an intensity of 1.50 parasites per individual; in Montes, prevalence was 31.88% with a parasite load of 6.82 parasites per host. Prevalence of metacercariae in crabs was 17. 98% in Montes and 4.07% in Sucre. No significant differences between prevalence, intensity and gender of the crabs was found in any of the two areas, but there was a positive and significant correlation with size. In Sucre, the crab parasite burden was higher, 8.20%, than in Montes, 5.31%. It was concluded that the intensity of infection was higher in female crabs and in those of a larger body size. These results indicate the existence of zoonotic contamination in both areas, and the need to continue investigating in order to determine relevant elements of this parasitic interrelation.

Broncoscopia intervencionista / Interventional bronchoscopy

La broncoscopia intervencionista ha presentado un gran desarrollo en la última década. Destacan los tratamientoscon láser traqueobronquial, la colocación de endoprótesis y la ecobroncoscopia. Se revisan las indicacionesy aplicaciones de estas 3 técnicas broncoscópicas(AU)

In the last decade, major advances have been made in interventional bronchoscopy. Notable techniques aretracheobronchial laser, endoprosthesis placement and endobronchial ultrasound bronchoscopy. Theindications and applications for these three bronchoscopic techniques are reviewed. I(AU)

[Validation of the SF-36 questionnaire in adults with asthma and allergic rhinitis in Mexican population]. / Validación del cuestionario SF-36 en pacientes adultos con asma y rinitis alérgica.

BACKGROUND: Multiple questionnaires demonstrated the effect of the disease on the Quality of Life (QL), especially on allergies. Asthma and allergic rhinitis contributes to the reduction of QL. One of the accepted instruments for the evaluation of QL in chronic diseases is the questionnaire SF-36 (SF-36). Our objective was to validate the SF-36 in Mexican asthmatic with allergic rhinitis adult patients. METHODS: Fifty asthmatic patients with allergic rhinitis participated. They were placed according to the severity of their asthma (GINA 2004) and of allergic rhinitis (ARIA). The SF-36 was applied. The results were analyzed by descriptive statistics and validation by internal consistency through the Cronbach's test. RESULTS: It was observed that greater deterioration of QL in asthmatic patients with allergic rhinitis is directly related with the severity of their disease. Greater deterioration was observed on physical nature and of the perception of health. The aspect less affected was the social function and mental health. The Cronbach's test showed a coefficient of global reliability of 0.9314. CONCLUSIONS: The results obtained for the validation of SF-36, through the Cronbach's test in the population studied allows their use for later studies.

[Interventional bronchoscopy]. / Broncoscopia intervencionista.

In the last decade, major advances have been made in interventional bronchoscopy. Notable techniques are tracheobronchial laser, endoprosthesis placement and endobronchial ultrasound bronchoscopy. The indications and applications for these three bronchoscopic techniques are reviewed. I.

Life cycle of Culuwiya tilapiae (Nasir y Gómez, 1976) (trematoda: haploporidae) in Venezuela

The life cycle of Culuwiya tilapiae (Nasir y Gómez, 1976) Overstreet and Curran, 2005, a digenean belonging to the family Haploporidae Nicoll, 1914 is redescibed and illustrated. The parasite infects naturally the snail, Pyrgophorus cf. spiralis (Guppy 1864), and the definitive host, Oreochromis mossambicus (Peters, 1852). Both molluscs and fish were collected from the Parque Litoral Laguna de los Patos, Cumaná, Estado Sucre, Venezuela. The cercariae were developed in redia in the digestive gland of molluscs and encysted on the water surface and more frequently on the bottom and walls of dishes. The 2-4 days old cysts of C. tilapiae were fed to laboratory reared O. mossambicus, and the trematode reaches sexual maturity in 22-32 days after the experimental infection. All developmental stages of C. tilapiae are described and illustrated. This species was compared with other members of Culuwiya and with Saccocoelioides tarpazensis Díaz and Gonzalez, 1990, for being the only recorded haploporids species in Venezuela and its life cycle had been elucidated and for the similarity with C. tilapiae. This species is redescribed herein represents the first record of Culuwiya Overstreet and Curran, 2005 in Venezuela.

El ciclo de vida de Culuwiya tilapiae (Nasir y Gómez, 1976) Overstreet y Curran, 2005, un digéneo perteneciente a la familia Haploporidae Nicoll, 1914 fue dilucidado. El parásito infecta naturalmente al caracol, Pyrgophorus cf. spiralis (Guppy, 1864), y al hospedador definitivo, Oreochromis mossambicus (Peters, 1852). Ambos, moluscos y peces fueron recolectados en el Parque Litoral Laguna de los Patos, Cumaná, estado Sucre, Venezuela. Las cercarias fueron desarrolladas en redias en la glándula digestiva del molusco, y enquista en la superficie del agua y más frecuente en el fondo y paredes del recipiente. Los quistes de 2-4 días de edad de C. tilapiae fueron suministrados a O. mossambicus nacidos en el laboratorio, y los tremátodos alcanzan su madurez sexual en 22-32 días después de la infección. Todos los estadios de desarrollo de C. tilapiae fueron descritos e ilustrados. Esta especie fue comparada con los miembros del género Culuwiya y con Saccocoelioides tarpazensis Díaz y González, 1990, por ser la única especie de haploporido en Venezuela, cuyo ciclo de vida ha sido dilucidado y por su similitud con C. tilapiae. Esta especie redescrita aquí, representa la primera vez que se registra el género en Venezuela.

[Pulmonary paragonimiasis. Case report]. / Paragonimiasis pulmonar. Descripción de un caso.

Paragonimiasis is a zoonosis affecting wild and domestic animals and human beings, caused by species of trematodes of the genus Paragonimus. Humans become infected after ingestion of raw or poorly cooked fresh water crustaceans. The aim of the present work is the description of a case of a 3-years old child, coming from Guárico State in Venezuela with a year of residence in the seashore of the Provincia Manabí in Ecuador, where he ate crabs in "ceviche". During hospitalization, he presented respiratory distress, hepatomegaly and nodules in the back. The thorax cat scan showed heavy infiltrate in both pulmonary bases and pleural compromise. Based on clinic, radiological images, an eosinophilia of 47% (Eosinophils absolute count (EAC) 6.682/mm3) and the antecedent of raw crabs ingestion, pulmonary paragonimiasis was diagnosed. Paragonimus eggs were not found in sputum and feces. ELISA with crude Paragonimus antigen was positive and Western blot revealed recognition of specific molecules. After treatment for three days with Praziquantel at a daily dose of 25 mg/kg body weight divided into three intakes, the sintomatology disappeared and radiological images and number of eosinophils diminished considerably.

Paragonimiasis pulmonar: descripción de un caso / Pulmonary paragonimiasis: case report

La Paragonimiasis es una zoonosis parasitaria de diversos animales silvestres o domésticos y del hombre, causada por especies de tremátodes del género Paragonimus siendo el Paragonimus westermani el que más infecta al hombre. El humano se infecta al ingerir crustáceos de agua dulce parasitados (cangrejos de río) crudos o insuficientemente cocidos. El objetivo de este trabajo es describir el caso de un preescolar masculino de 3 años y medio de edad, natural y procedente del estado Guárico quien vivió un año en zona costera del Ecuador (Provincia Manabí) donde consumió alimentos de la zona como ceviche. Consultó por dificultad respiratoria y durante la hospitalización se evidenció hepatomegalia y presencia de nódulos subcutáneos en espalda. La Tomografía Axial computarizada (TAC) de tórax reveló importante infiltrado a nivel de ambas bases pulmonares a predominio del lado derecho con derrame y engrosamiento pleural. Por la clínica, las imágenes radiológicas, la eosinofilia (47% con contaje absoluto de eosinófilos (CAE) 6.862/mm3) y el antecedente de ingesta de cangrejos crudos 6 meses antes, se sugirió descartar paragonimiasis pulmonar. En un estudio seriado de muestras de sueros se evidenció la presencia de anticuerpos específicos anti-Paragonimus por ELISA y Western blot, sin embargo no se encontraron huevos del parásito en heces o en esputo. Se indicó tratamiento con praziquantel 25 mg/kg de peso 3 tomas al día durante 3 días con lo cual desapareció la sintomatología, mejoraron las imágenes radiológicas y disminuyó el contaje de eosinófilos.