Functional genomics analysis of human colon organoids identifies key transcription factors.

Organoids are a valuable three-dimensional (3D) model to study the differentiated functions of the human intestinal epithelium. They are a particularly powerful tool to measure epithelial transport processes in health and disease. Though biological assays such as organoid swelling and intraluminal pH measurements are well established, their underlying functional genomics are not well characterized. Here we combine genome-wide analysis of open chromatin by ATAC-Seq with transcriptome mapping by RNA-Seq to define the genomic signature of human intestinal organoids (HIOs). These data provide an important tool for investigating key physiological and biochemical processes in the intestinal epithelium. We next compared the transcriptome and open chromatin profiles of HIOs with equivalent data sets from the Caco2 colorectal carcinoma line, which is an important two-dimensional (2D) model of the intestinal epithelium. Our results define common features of the intestinal epithelium in HIO and Caco2 and further illustrate the cancer-associated program of the cell line. Generation of Caco2 cysts enabled interrogation of the molecular divergence of the 2D and 3D cultures. Overrepresented motif analysis of open chromatin peaks identified caudal type homeobox 2 (CDX2) as a key activating transcription factor in HIO, but not in monolayer cultures of Caco2. However, the CDX2 motif becomes overrepresented in open chromatin from Caco2 cysts, reinforcing the importance of this factor in intestinal epithelial differentiation and function. Intersection of the HIO and Caco2 transcriptomes further showed functional overlap in pathways of ion transport and tight junction integrity, among others. These data contribute to understanding human intestinal organoid biology.

Inactivation of CFTR by CRISPR/Cas9 alters transcriptional regulation of inflammatory pathways and other networks.

BACKGROUND: Individuals with cystic fibrosis (CF) experience elevated inflammation in multiple organs, but whether this reflects an inherent feature of CF cells or is a consequence of a pro-inflammatory environment is not clear. METHOD: Using CRISPR/Cas9-mediated mutagenesis of CFTR, 17 subclonal cell lines were generated from Caco-2 cells. Clonal lines with functional CFTR (CFTR+) were compared to those without (CFTR-) to directly address the role of CFTR in inflammatory gene regulation. RESULTS: All lines maintained CFTR mRNA production and formation of tight junctions. CFTR+ lines displayed short circuit currents in response to forskolin, while the CFTR- lines did not. Baseline expression of cytokines IL6 and CXCL8 (IL8) was not different between the lines regardless of CFTR genotype. All lines responded to TNFα and IL1ß by increasing IL6 and CXCL8 mRNA levels, but the CFTR- lines produced more CXCL8 mRNA than the CFTR+ lines. Transcriptomes of 6 CFTR- and 6 CFTR+ lines, before and after stimulation by TNFα, were compared for differential expression as a function of CFTR genotype. While some genes appeared to be differentially expressed simply because of CFTR's absence, others required stimulation for differences to be apparent. CONCLUSION: Together, these data suggest cells respond to CFTR's absence by modulating transcriptional networks, some of which are only apparent when cells are exposed to different environmental contexts, such as inflammation. With regards to inflammation, these data suggest a model in which CFTR's absence leads to a poised, pro-inflammatory state of cells that is only revealed by stimulation.

Detección del gen fusión PML-RARα en pacientes colombianos con leucemia mieloide aguda / Detection of the PML-RARα fusion gene in Colombian patients with acute myeloid leukemia

Resumen Introducción: la leucemia promielocítica aguda es un subtipo de leucemia mieloide aguda caracterizada por la presencia de una translocación entre los cromosomas 15 y 17 que provoca la formación de un gen fusión denominado PML/RARα. Determinar la presencia de este gen fusión es crítico para estos pacientes ya que su presencia hace el diagnóstico de la enfermedad, aún sin tener resultados de patología. Con esta investigación se busca ajustar e implementar una prueba altamente sensible y específica para la detección del reordenamiento PML/RARα. Métodos: a partir de sangre periférica se extrajo RNA de pacientes diagnosticados con leucemia mieloide aguda en dos instituciones de Antioquia (Colombia). Se realizó RT-PCR anidada para la detección de PML/RARα, ajustando un protocolo previamente publicado. Resultados: se ajustó y estandarizó un método para detectar mediante RT-PCR el gen fusión PML/RARα. Mediante esta técnica se logró identificar la traslocación en cuatro pacientes (22 %) de la cohorte estudiada. Conclusiones: los resultados están de acuerdo con estudios previos. La detección de esta y otras alteraciones citogenéticas mediante pruebas moleculares permitirá tener información valiosa a nivel de diagnóstico y pronóstico de los pacientes con leucemia mieloide aguda en Antioquia.

Abstract Introduction: acute promyelocytic leukemia is a subtype of acute myeloid leukemia characterized by the presence of a translocation between chromosomes 15 and 17, which causes the formation of a fusion gene called PML/RARα. Determining the presence of this fusion gene is critical for these patients, since their presence makes the diagnosis of the disease, even with no pathology results This research seeks to adjust and implement a highly sensitive and specific test for the diagnosis of this cytogenetic abnormality. Methods: peripheral blood samples from patients diagnosed with acute myelocytic leukemia were collected in two institutions of Antioquia (Colombia), from which RNA was extracted and nested RT-PCR was performed, adjusting a previously published protocol. Results: we adjusted and standardized a method to detect the PML/RARα fusion gene by RT-PCR. Using this technique, translocation was identified in four patients (22%) of the studied cohort. Conclusions: our results agree with previous studies. The detection of this and other cytogenetic alterations by means of molecular tests will allow to have valuable information at the level of diagnosis and prognosis of patients with AML in Antioquia.

Acetyl-CoA carboxylase inhibition regulates microtubule dynamics and intracellular transport in cystic fibrosis epithelial cells.

The use of high-dose ibuprofen as an anti-inflammatory therapy in cystic fibrosis (CF) has been shown to be an effective intervention although use is limited due to potential adverse events. Identifying the mechanism of ibuprofen efficacy would aid in the development of new therapies that avoid these adverse events. Previous findings demonstrated that ibuprofen treatment restores the regulation of microtubule dynamics in CF epithelial cells through a 5'-adenosine monophosphate-activated protein kinase (AMPK)-dependent mechanism. The goal of this study is to define the AMPK pathway that leads to microtubule regulation. Here, it is identified that inhibition of acetyl-CoA carboxylase (ACC) is the key step in mediating the AMPK effect. ACC inhibition with 5-(tetradecyloxy)-2-furoic acid (TOFA) increases microtubule reformation rates in cultured and primary CF epithelial cells to wild-type (WT) rates. TOFA treatment also restores microtubule-dependent distribution of cholesterol and Rab7-positive organelles, as well as reduces expression of the proinflammatory signaling molecule RhoA to WT levels. ACC activation with citrate replicates these CF phenotypes in WT cells further supporting the role of AMPK signaling through ACC as a key mediator in CF cell signaling. It is concluded that ACC inhibition is the key step in the efficacy of AMPK activation at the cellular level and could represent a novel site of therapeutic intervention to address inflammation in CF.

Small adipose stores in cystic fibrosis mice are characterized by reduced cell volume, not cell number.

Cystic fibrosis (CF) is a lethal genetic disorder that affects many organ systems of the body, including various endocrine and exocrine tissues. Health and survival positively associate with body mass, and as a consequence, CF clinical care includes high-fat, high-calorie diets to maintain and increase adipose tissue stores. Such strategies have been implemented without a clear understanding of the cause and effect relationship between body mass and patients' health. Here, we used CF mouse models, which display small adipose stores, to begin examining body fat as a prelude into mechanistic studies of low body growth in CF, so that optimal therapeutic strategies could be developed. We reasoned that low adiposity must result from reduced number and/or volume of adipocytes. To determine relative contribution of either mechanism, we quantified volume of intraperitoneal and subcutaneous adipocytes. We found smaller, but not fewer, adipocytes in CF compared with wild-type (WT) animals. Specifically, intraperitoneal CF adipocytes were one-half the volume of WT cells, whereas subcutaneous cells were less affected by the Cftr genotype. No differences were found in cell types between CF and WT adipose tissues. Adipose tissue CFTR mRNA was detected, and we found greater CFTR expression in intraperitoneal depots as compared with subcutaneous samples. RNA sequencing revealed that CF adipose tissue exhibited lower expression of several key genes of adipocyte function ( Lep, Pck1, Fas, Jun), consistent with low triglyceride storage. The data indicate that CF adipocytes contain fewer triglycerides than WT cells, and a role for CFTR in these cells is proposed. NEW & NOTEWORTHY Adipocytes in cystic fibrosis mice exhibit smaller size due to low triglyceride storage. Adipocyte cell number per fat pad is similar, implying triglyceride storage problem. The absence of CFTR function in adipose tissue has been proposed as a direct link to low triglyceride storage in cystic fibrosis.

Absence of leptin signaling allows fat accretion in cystic fibrosis mice.

Negative energy balance is a prevalent feature of cystic fibrosis (CF). Pancreatic insufficiency, elevated energy expenditure, lung disease, and malnutrition, all characteristic of CF, contribute to the negative energy balance causing low body-growth phenotype. As low body weight and body mass index strongly correlate with poor lung health and survival of patients with CF, improving energy balance is an important clinical goal (e.g., high-fat diet). CF mouse models also exhibit negative energy balance (growth retardation and high energy expenditure), independent from exocrine pancreatic insufficiency, lung disease, and malnutrition. To improve energy balance through increased caloric intake and reduced energy expenditure, we disrupted leptin signaling by crossing the db/db leptin receptor allele with mice carrying the R117H Cftr mutation. Compared with db/db mice, absence of leptin signaling in CF mice (CF db/db) resulted in delayed and moderate hyperphagia with lower de novo lipogenesis and lipid deposition, producing only moderately obese CF mice. Greater body length was found in db/db mice but not in CF db/db, suggesting CF-dependent effect on bone growth. The db/db genotype resulted in lower energy expenditure regardless of Cftr genotype leading to obesity. Despite the db/db genotype, the CF genotype exhibited high respiratory quotient indicating elevated carbohydrate oxidation, thus limiting carbohydrates for lipogenesis. In summary, db/db-linked hyperphagia, elevated lipogenesis, and morbid obesity were partially suppressed by reduced CFTR activity. CF mice still accrued large amounts of adipose tissue in contrast to mice fed a high-fat diet, thus highlighting the importance of dietary carbohydrates and not simply fat for energy balance in CF. NEW & NOTEWORTHY We show that cystic fibrosis (CF) mice are able to accrue fat under conditions of carbohydrate overfeeding, increased lipogenesis, and decreased energy expenditure, although length was unaffected. High-fat diet feeding failed to improve growth in CF mice. Morbid db/db-like obesity was reduced in CF double-mutant mice by reduced CFTR activity.

Molecular characterization of gene regulatory networks in primary human tracheal and bronchial epithelial cells.

BACKGROUND: Robust methods to culture primary airway epithelial cells were developed several decades ago and these cells provide the model of choice to investigate many diseases of the human lung. However, the molecular signature of cells from different regions of the airway epithelium has not been well characterized. METHODS: We utilize DNase-seq and RNA-seq to examine the molecular signatures of primary cells derived from human tracheal and bronchial tissues, as well as healthy and diseased (cystic fibrosis (CF)) donor lung tissue. RESULTS: Our data reveal an airway cell signature that is divergent from other epithelial cell types and from common airway epithelial cell lines. The differences between tracheal and bronchial cells are clearly evident as are common regulatory features. Only minor variation is seen between bronchial cells from healthy or CF donors. CONCLUSIONS: These data are a valuable resource for functional genomics analysis of airway epithelial tissues in human disease.

Prevalencia y factores de riesgo asociados a alteraciones comunicativas en vendedores ambulantes de Popayán, Colombia / Communicative disorder prevalence and associated risk factors regarding informal workers in Popayan, Colombia

Objetivo Determinar la prevalencia de alteraciones en la audición, función respiratoria y vocal y su asociación con ciertos factores de riesgo en vendedores ambulantes de la ciudad de Popayán, Colombia. Métodos Estudio descriptivo de corte transversal en 186 vendedores ambulantes. Después de la firma del consentimiento informado, se aplicó una encuesta para obtener datos sobre variables sociodemográficas y comunicativas. Posteriormente, los trabajadores fueron evaluados mediante la aplicación de pruebas de audiometría tonal, funcionalidad respiratoria y perfil vocal de Wilson. El análisis estadístico se realizó en SPSS v. 19.0. Resultados Ser mayor de 30 años (OR 5,84; IC95 % 2,85-12,00), poseer nivel educativo (2,81; 1,22-6,44) y estrato socioeconómico bajos (4,54; 1,89-10,91), y el tiempo de trabajo prolongado (2,64; 1,27-5,06) estuvieron asociados a alteraciones en la función auditiva. Alteraciones en la función respiratoria estuvieron asociadas a ser mujer (1,83; 1,00-3,34) y tiempo de trabajo prolongado (2,04; 1,11-3,74). Para la función vocal, ser mayor de 30 años (3,36; IC 1,33-3,51) y el nivel educativo bajo (3,67; 1,05-12,76) fueron factores de riesgo. Los antecedentes comunicativos relacionados con alteraciones auditivas, respiratorias y de la voz fueron dolor de oído, hipertrofia de amígdalas, trauma, reflujo gastroesofágico, tos frecuente, emociones fuertes y gritar. Conclusiones Se evidencia que las alteraciones en el sistema comunicativo en los vendedores ambulantes está asociada con ciertos factores de riesgo sociodemográficos y comunicativos. Esta información constituye una línea de base para mejorar las estrategias de promoción de la salud y prevención de la enfermedad en esta población.(AU)

Objective Determining the prevalence of alterations in informal workers’ audition, respiratory and vocal functions and their association with certain risk factors in Popayan, Colombia. Methods This was a descriptive, cross-sectional study of 186 informal workers (i.e. people selling things in the street). After signing an informed consent form, an interview was held to obtain data regarding sociodemographic and communicative variables. The workers were then evaluated using tests for tone audiometry, respiratory function and Wilson's voice profile. SPSS (v.19.0) was used for statistical analysis. Results Being older than 30 years of age (OR 5.84: 2.85-12.00 95%CI), having a poor educational level (2.81: 1.22-6.44 95%CI) and low socioeconomic status (4.54:1.89-10.91 95%CI) and prolonged working hours (2.64: 1.27-5.06 95%CI) were associated with auditive function disorders. Respiratory function disorders were associated with being female (1.83; 1.00-3.34 95%CI) and having prolonged working hours (2.04: 1.11-3.74 95%CI). Regarding vocal function, being over 30 years-old (3.36: 1.33-3.51 95%CI) and having a low educational level (3.67; 1.05-12.76 95%CI) were risk factors. Communicative factors related to auditive, respiratory and voice disorder were ear pain, hypertrophic tonsils, trauma, gastro-esophageal reflux, frequent cough, strong emotions and screaming. Conclusions It is evident that alterations in informal workers’ communicative system are associated with certain sociodemographic and communicative risk factors. This information provides a baseline for improving healthcare promotion and disease prevention strategies aimed at this population.(AU)

Caracterización de la intoxicación exógena en niños y adolescentes en Sogamoso, Boyacá durante el período de 2010 a 2013 / Characterization of exogenous poisoning in children and teenagers at Sogamoso, Boyacá, during the period 2010 to 2013

Introducción: las intoxicaciones exógenas han adquirido paulatinamente la condición de problema de salud pública, particularmente en el área pediátrica, dado que son causantes de hasta un 10% de todos los ingresos a la Unidad de Cuidado Intensivo pediátrica. Objetivo: caracterizar los patrones epidemiológicos generales de la intoxicación exógena aguda ocurrida en niños y adolescentes de menos de 18 años, ingresados al servicio de urgencias del Hospital Regional de Sogamoso, Boyacá, durante el período comprendido entre enero de 2010 y junio de 2013. Materiales y Métodos: estudio observacional descriptivo. Se realizó una revisión sistemática de 143 historias clínicas de menores de 18 años ingresados al servicio de urgencias con diagnóstico de intoxicación exógena; se evaluaron variables como edad, género, agente, vía de administración, motivo, sitio de ocurrencia y desenlace. Resultados: media de edad 10,5 años (SD=5,6); 56,6% de los casos ocurrieron en pacientes de sexo femenino. Se identificaron dos patrones temporales de intoxicación: uno accidental en menores de cinco años y otro con motivación suicida en adolescentes, más frecuente en mujeres. La mayoría de eventos ocurrieron en casa y las sustancias más frecuentemente involucradas fueron medicamentos y productos de higiene y aseo como plaguicidas y desinfectantes; alta prevalencia de intoxicación accidental por alimentos contaminados; en 5,9% de los casos se encontró como comorbilidad el abuso sexual. Conclusiones: es recomendable el diseño e implementación de acciones educativas dirigidas a padres y cuidadores, así como la de intervenciones de vigilancia y control de situaciones propiciadoras de la conducta suicida como los estados depresivos y el estrés en adolescentes. (MÉD.UIS. 2014;27(1):9-15).

Introduction: exogenous poisoning has gradually acquired the status of a public health problem, particularly in the pediatric area, since they are causing up to 10% of all admissions to the Pediatric Intensive Care Unit. Objective: to characterize the general epidemiological patterns of acute exogenous poisoning, occurred in children and teenagers aged less than 18 years, admitted to the emergency department of the Hospital Regional de Sogamoso, Boyacá, during the period between january 2010 and june 2013. Materials and Methods: descriptive observational study. We conducted a systematic review of 143 medical records of children aged less than 18 years, admitted to the emergency department with a diagnosis of exogenous intoxication. There were evaluated variables such as age, gender, agent, route of administration, motive, site of occurrence and final outcome. Results: mean age 10.5 years (SD = 5.6), 56.6% of cases occurred in girls. It was present two temporal patterns of intoxication, one accidental in children aged equal or less than five years and another motivated by suicidal ideation in teenagers, more common in female. Most events occurred at home and the substances most frequently involved were medicaments and hygiene products and cleaning as pesticides and disinfectants. There was high prevalence of accidental poisoning from contaminated food. In 5.9% of cases was found sexual abuse comorbidity. Conclusions: it's recommendable the design and implementation of educational activities aimed to parents and caretakers, as well as monitoring and control interventions on cases with propitiating situations of suicidal ideation such as depression and stress in teenagers. (MÉD.UIS. 2014;27(1):9-15).

[Communicative disorder prevalence and associated risk factors regarding informal workers in Popayan, Colombia]. / Prevalencia y factores de riesgo asociados a alteraciones comunicativas en vendedores ambulantes de Popayán, Colombia.

OBJECTIVE: Determining the prevalence of alterations in informal workers’ audition, respiratory and vocal functions and their association with certain risk factors in Popayan, Colombia. METHODS: This was a descriptive, cross-sectional study of 186 informal workers (i.e. people selling things in the street). After signing an informed consent form, an interview was held to obtain data regarding sociodemographic and communicative variables. The workers were then evaluated using tests for tone audiometry, respiratory function and Wilson's voice profile. SPSS (v.19.0) was used for statistical analysis. RESULTS: Being older than 30 years of age (OR 5.84: 2.85-12.00 95%CI), having a poor educational level (2.81: 1.22-6.44 95%CI) and low socioeconomic status (4.54:1.89-10.91 95%CI) and prolonged working hours (2.64: 1.27-5.06 95%CI) were associated with auditive function disorders. Respiratory function disorders were associated with being female (1.83; 1.00-3.34 95%CI) and having prolonged working hours (2.04: 1.11-3.74 95%CI). Regarding vocal function, being over 30 years-old (3.36: 1.33-3.51 95%CI) and having a low educational level (3.67; 1.05-12.76 95%CI) were risk factors. Communicative factors related to auditive, respiratory and voice disorder were ear pain, hypertrophic tonsils, trauma, gastro-esophageal reflux, frequent cough, strong emotions and screaming. CONCLUSIONS: It is evident that alterations in informal workers'’ communicative system are associated with certain sociodemographic and communicative risk factors. This information provides a baseline for improving healthcare promotion and disease prevention strategies aimed at this population.

Altered de novo lipogenesis contributes to low adipose stores in cystic fibrosis mice.

Cystic fibrosis (CF) mouse models exhibit exocrine pancreatic function, yet they do not develop adipose stores to the levels of non-CF mice. CF mice homozygous for the Cftr mutation (F508del) at 3 wk (postweaning) and 6 wk (young adult) of age had markedly less adipose tissue than non-CF mice. Food intake was markedly lower in 3-wk-old CF mice but normalized by 6 wk of age. Both 3- and 6-wk-old mice had dietary lipid absorption and fecal lipid excretion comparable to non-CF mice. Hepatic de novo lipogenesis (DNL), determined by (2)H incorporation, was reduced in CF mice. At 3 wk, F508del mice had significantly decreased DNL of palmitate and stearate, by 83% and 80%, respectively. By 6 wk, DNL rates in non-CF mice remained unchanged compared with 3-wk-old mice, while DNL rates of F508del mice were still reduced, by 33% and 40%, respectively. Adipose tissue fatty acid (FA) profiles were comparable in CF and non-CF mice, indicating that adipose differences are quantitative, not qualitative. A correspondingly lower content of (2)H-labeled FA was found in CF adipose tissue, consistent with reduced deposition of newly made hepatic triglycerides and/or decreased adipose tissue lipogenesis. Hepatic transcriptome analysis revealed lower mRNA expression from several genes involved in FA biosynthesis, suggesting downregulation of this pathway as a mechanism for the reduced lipogenesis. These novel data provide a model for altered lipid metabolism in CF, independent of malabsorption, and may partly explain the inability of pancreatic enzyme replacement therapy to completely restore normal body mass to CF patients.

Peroxisome proliferator-activated receptor-gamma in cystic fibrosis lung epithelium.

The pathophysiology of cystic fibrosis (CF) inflammatory lung disease is not well understood. CF airway epithelial cells respond to inflammatory stimuli with increased production of proinflammatory cytokines as a result of increased NF-kappaB activation. Peroxisome proliferator-activated receptor-gamma (PPARgamma) inhibits NF-kappaB activity and is reported to be reduced in CF. If PPARgamma participates in regulatory dysfunction in the CF lung, perhaps PPARgamma ligands might be useful therapeutically. Cell models of CF airway epithelium were used to evaluate PPARgamma expression and binding to NF-kappaB at basal and under conditions of inflammatory stimulation by Pseudomonas aeruginosa or TNFalpha/IL-1beta. An animal model of CF was used to evaluate the potential of PPARgamma agonists as therapeutic agents in vivo. In vitro, PPARgamma agonists reduced IL-8 and MMP-9 release from airway epithelial cells in response to PAO1 or TNFalpha/IL-1beta stimulation. Less NF-kappaB bound to PPARgamma in CF than normal cells, in two different assays; PPARgamma agonists abrogated this reduction. PPARgamma bound less to its target DNA sequence in CF cells. To test the importance of the reported PPARgamma inactivation by phosphorylation, we observed that inhibitors of ERK, but not JNK, were synergistic with PPARgamma agonists in reducing IL-8 secretion. In vivo, administration of PPARgamma agonists reduced airway inflammation in response to acute infection with P. aeruginosa in CF, but not wild-type, mice. In summary, PPARgamma inhibits the inflammatory response in CF, at least in part by interaction with NF-kappaB in airway epithelial cells. PPARgamma agonists may be therapeutic in CF.

CFTR inhibition mimics the cystic fibrosis inflammatory profile.

Primary airway epithelial cells grown in air-liquid interface differentiate into cultures that resemble native epithelium morphologically, express ion transport similar to those in vivo, and secrete cytokines in response to stimuli. Comparisons of cultures derived from normal and cystic fibrosis (CF) individuals are difficult to interpret due to genetic differences besides CFTR. The recently discovered CFTR inhibitor, CFTR(inh)-172, was used to create a CF model with its own control to test if loss of CFTR-Cl(-) conductance alone was sufficient to initiate the CF inflammatory response. Continuous inhibition of CFTR-Cl(-) conductance for 3-5 days resulted in significant increase in IL-8 secretion at basal (P = 0.006) and in response to 10(9) Pseudomonas (P = 0.0001), a fourfold decrease in Smad3 expression (P = 0.02), a threefold increase in RhoA expression, and increased NF-kappaB nuclear translocation upon TNF-alpha/IL-1beta stimulation (P < 0.000001). CFTR inhibition by CFTR(inh)-172 over this period does not increase epithelial sodium channel activity, so lack of Cl(-) conductance alone can mimic the inflammatory CF phenotype. CFTR(inh)-172 does not affect IL-8, IL-6, or granulocyte/macrophage colony-stimulating factor secretion in two CF phenotype immortalized cell lines: 9/HTEo(-) pCEP-R and 16HBE14o(-) AS, or IL-8 secretion in primary CF cells, and inhibitor withdrawal abolishes the increased response, so CFTR(inh)-172 effects on cytokines are not direct. Five-day treatment with CFTR(inh)-172 does not affect cells deleteriously as evidenced by lactate dehydrogenase, trypan blue, ciliary activity, electron micrograph histology, and inhibition reversibility. Our results support the hypothesis that lack of CFTR activity is responsible for the onset of the inflammatory cascade in the CF lung.

Antibodies to the polymeric immunoglobulin receptor with different binding and trafficking patterns.

The polymeric immunoglobulin receptor (pIgR) has been proposed as a therapeutic target, but its potential depends on the efficiency of uptake and trafficking of the receptor ligand. Mouse monoclonal antibodies (Mabs) directed against pIgR, selected for strong binding to secretory component (SC) and secretory IgA (sIgA), were tested in a transcytosis assay in 16HBEo--cells (human bronchial epithelial cell line) transfected with human pIgR. Intracellular trafficking was followed by confocal microscopy. Mabs fell into two classes. For two Mabs, transcytosis from basolateral to apical surface is rapid, unidirectional, and little Mab is retained in the cell. For three Mabs, basolateral to apical transcytosis occurs to a significantly lesser extent, reverse transcytosis is permitted, and some of the Mab is retained in the perinuclear region even after 24 h. When tested for their ability to recognize and immunoprecipitate pIgR with systematic truncations and deletions of the five immunoglobulin (Ig)-like domains, all Mabs bound to the fifth Ig-like domain, but three of them also bound to the C-terminal region of pIgR near the plasma membrane. Different binding sites probably account for the different trafficking of these Mabs and may predict differential therapeutic utility.

Gene profile changes after Pseudomonas aeruginosa exposure in immortalized airway epithelial cells.

To test the hypothesis that the excess inflammatory response in cystic fibrosis airway epithelial cells is the result of differential activation of genes in response to a laboratory strain of Pseudomonas aeruginosa (PAO1), a 48-h time course of genes expressed following PAO1 stimulation (10(9) CFU for 1 h) was studied in two pairs of airway epithelial cells: 9/HTEo- and 16HBE14o-, each with a matched normal and CF phenotype pair. cRNA was hybridized to Affymetrix HG-U95Av2 chips and pairwise comparisons against zero time (no PAO1) were calculated for each time point. PAO1 elicited profound changes in both cell pairs: for 9/HTEo-, 144 genes changed significantly in the normal pair, and 116 for the CF pair; for the 16HBE14o- pair, 57 genes changed significantly for the normal pair and 53 for the CF pair. Changes were much greater in the 9/HTEo- than in the 16HBE14o- pair, but basal levels of expression of inflammatory genes are higher in the 16HBE14o- pair, so 16HBE14o- was used mainly to corroborate the results of 9/HTEo-. Clustering analysis indicated that the pattern of gene expression is similar in the CF cells and their normal counterparts. However, there were substantial quantitative differences in gene expression. Thus, the difference between CF and normal resides in the magnitude, not the pattern, of the changes.

Calidad de atención en el puerperio inmediato del Hospital Universitario del Valle, Cali, 1993

Se realizó un estudio descriptivo-prospectivo con una muestra de 114 mujeres que se observaron desde el ingreso al servicio de puerperio hasta su egreso, para evaluar la calidad de atención que se brinda en el puerperio inmediato en el Hospital Universitario del Valle, Cali, Colombia, durante los meses de octubre a diciembre de 1993. Se evaluaron tres componentes: el científico-técnico que permitió analizar el cumplimiento de normas, la existencia de manuales, recurso humano y material, el de ambiente físico que permitió determinar las condiciones del área en donde se presta la atención y por último el componente interpersonal que permitió identificar la opinión de la usuaria acerca de la atención humanizada recibida. El servicio de puerperio cumplió con el 63.5 por ciento de los requisitos mínimos establecidos para recursos humanos, con un 68.3 por ciento para recursos materiales, con un 18.2 por ciento para el cumplimiento de normas y un 66.6 por ciento para la existencia de manuales. El 68.8 por ciento de las mujeres opinó que la atención recibida fue humanizada, las condiciones del ambiente físico obtuvo un 56.7 por ciento de cumplimiento de los requisitos mínimos establecidos. Estos resultados reportados que ningún componente obtuvo un porcentaje mayor del 80 por ciento(porcentaje mínimo establecido), lo que índico que la calidad de atención en el puerperio inmediato no fue adecuado

Sensibilización por primo vacunación BCG en lactantes sanos mediante la aplicación de PPD / Sensitivity by prime vaccination BCG in healthy infants by means the application of PPD

La vacuna BCG se viene empleando en Venezuela desde hace aproximadamente medio siglo, no obstante, existen escasos datos sobre su sensibilización detectada con PPD en nuestros niños. Un estudio previo de población escolar vacunada múltiples veces con BCG, arrojó una muy escasa sensibilización (0.5 por ciento con induración mayor de 10mm). Es por razón, que nos propusimos estudiar una muestra de 45 lactantes sanos primo-vacunados al nacer, empleando un método de lectura estandarizado a las 72 horas, con un PPD biológicamente activo (2uT, RT-23, suministrado por el MSAS). Los resultados confirman la baja sensibilización encontrada previamente en escolars, ya que en un 62 por ciento de la muestra no se detectó induración alguna; solamente un 4,4 por ciento presentó criterio de induración mayor de 10mm. En la gran mayoría de nuestros lactantes (80 por ciento ) se pudo apreciar cicatriz de BCG (promedio 4mm), indicando una adecuada inoculación. Queremos estimular la realización de estudios similares en distintas regiones del país, pensando en la posibilidad de que si revelan resultados comparables, un escrutinio del programa nacional de control de la tuberculosis en lo que respecta a la primo-vacunación BCG, sería recomendable

Soporte nutricional por microyeyunostomía en embarazo de 19 semanas llevado a término: primer caso en Venezuela / Nutritional support for microjejunostomy in a pregnancy of 19 weeks terminated: first case in Venezuela

Se describe un caso de esófago gastritis cáustica y estenosis pilórica, consecuente a un intento de suicidio en paciente de 26 años y embarazo de 19 semanas. Se trató con nutrición parenteral y enteral, vía microyeyunostomía y cirugía abdominal logrando la viabilidad fetal y restablecimiento de la salud materna