Posibilidad de terapia génica en pacientes con enfermedades retinianas hereditarias / Possibility of genetic therapy for inherited retinal conditions

Objetivo Evaluar la posibilidad de terapia génica en pacientes con enfermedades oculares hereditarias con diagnóstico genético establecido. Los objetivos secundarios son revisar la tasa de diagnóstico genético y hacer una actualización de los genes para los cuales hay estudios clínicos o preclínicos en curso que pudieran permitir la terapia génica. Métodos Estudio observacional, retrospectivo y multicéntrico de 177 pacientes con enfermedades oculares hereditarias a quienes se realizó estudio genético.Resultados De 177 pacientes con estudio genético, se incluyeron 146. Se identificaron variantes causantes de enfermedad en 117 pacientes con lo que se obtuvo una tasa de detección de variantes del 80,1%. Se encontraron variantes patogénicas en 47 genes, siendo ABCA4 el gen más común (17,9%), seguido por CRB1 (11,9%). De los pacientes con diagnóstico genético, el 64,1% tienen una variante en un gen para el cual se ha estudiado terapia génica y solo el 40,1% presentan una variante en genes con estudios para su terapia génica en fase clínica. Conclusiones El estudio genético ha abierto nuevos horizontes en el manejo de pacientes con enfermedades oculares hereditarias. Cerca de dos tercios de los pacientes presentó variantes patogénicas en genes para los cuales se ha evaluado la posibilidad de terapia génica. Sin embargo, muchos estudios se encuentran en fase preclínica. Se debe adecuar las expectativas de los pacientes sometidos a estudio genético y sus familias (AU)

Objective To evaluate the possibility of gene therapy in patients with inherited ocular conditions and established genetic diagnosis. The secondary objectives were to determine the genetic diagnostic rate and to update the list of genes for which there are ongoing clinical trials or preclinical studies that could allow for gene therapy. Methods Observational, retrospective, multicentric study of 177 patients with inherited ocular conditions that underwent genetic testing. Results Of 177 patients with genetic testing, 146 were enrolled for this study. Disease-causing variants were identified in 117 patients (variant detection rate of 80.1%). Pathogenic variants were found in 47 genes, with ABCA4 being the most common gene (17.9%), followed by CRB1 (11.9%). 64.1% of patients with a genetic diagnosis have a variant in genes for which gene therapy has been studied and only 40.1% have a variant in genes with studies for gene therapy in clinical phase. Conclusions Genetic testing has opened new horizons in the management of patients with hereditary ocular diseases. About two-thirds of the patients had pathogenic variants in genes for which gene therapy has been evaluated. However, many studies are in the pre-clinical phase. The expectations of patients undergoing genetic study and their families should be managed accordingly (AU)

Possibility of genetic therapy for inherited retinal conditions.

OBJECTIVE: To evaluate the possibility of gene therapy in patients with inherited ocular conditions and established genetic diagnosis. The secondary objectives were to determine the genetic diagnostic rate and to update the list of genes for which there are ongoing clinical trials or preclinical studies that could allow for gene therapy. METHODS: Observational, retrospective, multicentric study of 177 patients with inherited ocular conditions that underwent genetic testing. RESULTS: Of 177 patients with genetic testing, 146 were enrolled for this study. Disease-causing variants were identified in 117 patients (variant detection rate of 80.1%). Pathogenic variants were found in 47 genes, with ABCA4 being the most common gene (17.9%), followed by CRB1 (11.9%). 64.1% of patients with a genetic diagnosis have a variant in genes for which gene therapy has been studied and only 40.1% have a variant in genes with studies for gene therapy in clinical phase. CONCLUSIONS: Genetic testing has opened new horizons in the management of patients with hereditary ocular diseases. About two-thirds of the patients had pathogenic variants in genes for which gene therapy has been evaluated. However, many studies are in the pre-clinical phase. The expectations of patients undergoing genetic study and their families should be managed accordingly.

Utilidad de la quimioterapia con corticoides en la reducción de volumen de las lesiones en fetos con malformación congénita de la vía aérea pulmonar (MCVAP): serie de casos / Usefulness of chemotherapy with corticosteroids in reducing the volume of the lesions in fetuses with congenital pulmonary airway malformation (MCVAP): case series

RESUMEN OBJETIVO: el propósito es describir el efecto de los corticoides prenatales en la malformación congénita de la vía aérea pulmonar (MCVAP) mediante la evaluación secuencial del CVR y mostrar la experiencia en nuestro centro de terapia fetal. MATERIALES Y MÉTODOS: serie de casos en la cual se incluyeron ocho pacientes del Hospital de San José; con diagnóstico de MCVAP; se evaluó edad gestacional, el CVR (longitud x largo x ancho, en cms de la masa) x 0.523/circunferencia cefálica) como factor pronóstico, la desviación mediastínica, la presencia de hidrops, el tipo de MCVAP y la localización. Se aplicó betametasona a dosis de inducción de madurez pulmonar fetal. Se realizó la medición del CVR pre y pos tratamiento. Los datos obtenidos fueron analizados mediante el software estadístico SPSS (Versión 11.0 SPSS Inc. Chicago, IL). Se utilizaron la prueba Wilcoxon y el índice de correlación de Spearman según el caso. Un valor de p<0.05 se consideró significativo. RESULTADOS: ocho pacientes fueron incluidas. La mediana de la edad gestacional fue 30.5 semanas. Cuatro casos fueron tipo I, dos tipo II y dos tipo tres. Cuatro casos presentaron desviación mediastinal. Cinco casos fueron derechos y uno bilateral. Ningún caso presentó hidrops. El CVR pre tratamiento fue 0,93(RIQ:0,17-2,1) y pos tratamiento 0,55(RIQ:0,07-1,39). Se encontró una disminución del CVR pos tratamiento en todos los casos (p 0.0117) e índice de correlación de Spearman 0.9524 CONCLUSIÓN: Después de la aplicación de corticoides encontramos una disminución del CVR en todos los casos descritos. El uso de corticoides podría ser parte de la terapia prenatal con miras a mejorar el pronóstico.

ABSTRACT OBJECTIVE: The purpose of this study is to describe the effect of prenatal corticosteroids in the CPAM by sequential evaluation of the CVR, and to show the experience in our fetal therapy center. MATERIALS AND METHODS: Series of cases in which eight patients from the Hospital of San José were included; with diagnosis of CPAM; gestational age was evaluated, CVR (CVR = length x length x width cm, mass) x 0.523 / head circumference) as a prognostic factor, mediastinal shift, hydrops, type of CPAM, location. Betamethasone was applied to fetal lung maturation dose. In all patients underwent CVR measurement pre and post treatment. The data were analyzed using SPSS statistical software (Version 11.0 SPSS Inc. Chicago, IL). The Wilcoxon test and the Spearman correlation index were used according to the case. A value of p <0.05 was considered significant. RESULTS: eight patients were included. The median of gestational age was 30.5 weeks. Four cases were type I, two type II and two type three. Four cases were mediastinal shift. Five cases were rights and one bilateral. No case presented hydrops. The pretreatment CVR was 0,93(RIQ: 0,17-2,1) and post-treatment 0,55(RIQ: 0,07-1,39). We found a decrease in CVR after treatment in all cases (p 0.0117) and Spearman correlation index 0.9524 CONCLUSION: After application of corticosteroids are a reduction in CVR in all the cases described. The use of corticosteroids may be part of prenatal therapy to improve prognosis.

Differential gene expression in porcine oviduct during the oestrous cycle.

The oviduct undergoes changes under the influence of steroid hormones during the oestrous cycle. However, the molecular mechanisms underlying oviductal regulation are not fully understood. The aim of the present study was to identify the gene expression profile of the porcine oviduct in different stages of the cycle using microarray technology. A systematic study was performed on animals at four different stage: prepubertal gilts, and sows in the preovulatory, postovulatory and luteal phase of the oestrous cycle. The porcine oviduct expressed a total of 4929 genes. Moreover, significant differences in the expression of several genes were detected as the oestrous cycle progressed. Analysis of the differentially expressed genes indicated that a total of 86, 89 and 15 genes were upregulated in prepubertal gilts, preovulatory and luteal sows respectively compared with levels observed in postovulatory sows. Moreover, 80, 51 and 64 genes were downregulated in prepubertal, preovulatory and luteal animals respectively compared with the postovulatory sows. The concentrations of 10 selected transcripts were quantified by real-time reverse transcription-polymerase chain reaction to validate the cDNA array hybridisation data. Conversely, for some genes, localisation of corresponding protein expression in the oviduct was analysed by immunohistochemistry (i.e. cholecystokinin, glutathione peroxidase 2, mucin 1, phosphatidylethanolamine binding protein 4 and tachykinin 3) and mass spectrometry analysis of oviductal fluid allowed identification of peptides from all five proteins. The results of the present study demonstrate that gene expression in the porcine oviduct is clearly regulated during the oestrous cycle, with some oviductal proteins that could be related to several reproductive processes described here for the first time.

Synthesis and Complete Antimicrobial Characterization of CEOBACTER, an Ag-Based Nanocomposite.

The antimicrobial activity of silver nanoparticles (AgNPs) is currently used as an alternative disinfectant with diverse applications, ranging from decontamination of aquatic environments to disinfection of medical devices and instrumentation. However, incorporation of AgNPs to the environment causes collateral damage that should be avoided. In this work, a novel Ag-based nanocomposite (CEOBACTER) was successfully synthetized. It showed excellent antimicrobial properties without the spread of AgNPs into the environment. The complete CEOBACTER antimicrobial characterization protocol is presented herein. It is straightforward and reproducible and could be considered for the systematic characterization of antimicrobial nanomaterials. CEOBACTER showed minimal bactericidal concentration of 3 µg/ml, bactericidal action time of 2 hours and re-use capacity of at least five times against E. coli cultures. The bactericidal mechanism is the release of Ag ions. CEOBACTER displays potent bactericidal properties, long lifetime, high stability and re-use capacity, and it does not dissolve in the solution. These characteristics point to its potential use as a bactericidal agent for decontamination of aqueous environments.

Eye-tracking capabilities of low-cost EOG system.

This paper presents the design and implementation of a low-cost eye tracking system that allows measuring the rotational angle of the eye and gaze direction in healthy individuals. The system consists of an EOG circuit with simple components that acquire both horizontal and vertical eye movement through regular all-purpose contact electrodes. Then the data are analyzed and translated into corresponding angle values representing the eye rotation angle in both orientations. Results show that horizontal angle measurements are much more accurate than vertical measurements. A discussion regarding the performance and possible improvements is presented.

Management of subluxated lens in young patients.

OBJECTIVES: To evaluate visual outcomes in patients treated for lens subluxation. Secondary objectives are to report best corrected visual acuity (BCVA) in LogMAR and compare the outcomes of patients managed conservatively with those treated surgically. METHODS: Retrospective comparison of BCVA in patients under 50 years-old with lens subluxation, managed conservatively or surgically. RESULTS: A total of 49 eyes of 28 patients were included. Demographic characteristics were similar in both groups. Twenty eyes were treated surgically (40.8%) compared to 29 with medical treatment (59.2%). Marfan syndrome (79.6%) was diagnosed in 39 eyes. LogMAR BCVA post intervention was 0.35±0.31 for medical treatment and 0.39±0.32 for the surgical group, with no significant differences (P=.63). Improvements in LogMAR lines were 2.7±4.2 and 4.11±4.2 (P=.35), respectively. Two eyes in the surgery group developed ocular hypertension (0.04%), none with retinal detachment. CONCLUSIONS: The final BCVA showed no significant differences in this group of patients. BCVA depends on the visual potential of the rehabilitated eye rather than a specific type of intervention.

Manejo del cristalino subluxado en pacientes jóvenes / Management of subluxated lens in young patients

Objetivos: Evaluar los resultados visuales en pacientes tratados por subluxación del cristalino. Los objetivos secundarios son reportar la agudeza mejor corregida (AVMC) en LogMAR y comparar los resultados de los pacientes manejados de manera conservadora con aquellos manejados quirúrgicamente. Métodos: Estudio retrospectivo comparativo de AVMC en pacientes menores de 50 años con subluxación de cristalino, manejados de manera conservadora con lentes o tratados quirúrgicamente. Resultados: Se incluyeron 49 ojos de 28 pacientes. Las características demográficas fueron similares en ambos grupos; 20 ojos fueron tratados quirúrgicamente (40,8%) versus 29 con tratamiento médico (59,2%); 39 ojos con diagnóstico de síndrome de Marfán (79,6%). La AVMC LogMAR postintervención de 0,35±0,31 para el tratamiento médico y 0,39±0,32 para el grupo de manejo quirúrgico, sin encontrarse diferencias significativas (p=0,63). Las mejorías en líneas LogMAR fueron, respectivamente, 2,7±4,2 y 4,11±4,2 (p=0,35). Dos ojos del grupo cirugía evolucionaron con hipertensión ocular (0,04%), ninguno con DR. Conclusiones: La AVMC final no presentó diferencias significativas en este grupo de pacientes, dependiendo del potencial visual del ojo rehabilitado más que de un tipo específico de intervención(AU)

Objectives: To evaluate visual outcomes in patients treated for lens subluxation. Secondary objectives are to report best corrected visual acuity (BCVA) in LogMAR and compare the outcomes of patients managed conservatively with those treated surgically. Methods: Retrospective comparison of BCVA in patients under 50 years-old with lens subluxation, managed conservatively or surgically. Results: A total of 49 eyes of 28 patients were included. Demographic characteristics were similar in both groups. Twenty eyes were treated surgically (40.8%) compared to 29 with medical treatment (59.2%). Marfan syndrome (79.6%) was diagnosed in 39 eyes. LogMAR BCVA post intervention was 0.35±0.31 for medical treatment and 0.39±0.32 for the surgical group, with no significant differences (P=.63). Improvements in LogMAR lines were 2.7±4.2 and 4.11±4.2 (P=0.35), respectively. Two eyes in the surgery group developed ocular hypertension (0.04%), none with retinal detachment. Conclusions: The final BCVA showed no significant differences in this group of patients. BCVA depends on the visual potential of the rehabilitated eye rather than a specific type of intervention(AU)

The oviduct: functional genomic and proteomic approach.

The mammalian oviduct is an anatomical part of the female reproductive tract, which plays several important roles in the events related to fertilization and embryo development. This review examines and compares several studies related to the proteomic and transcriptomic profile of the oviduct in different domestic animals. This information could be important for clarifying the role of oviductal factors in different events regulating fertilization and early embryo development, as well as for improving synthetic media for in vitro maturation/in vitro fertilization/embryo culture techniques (IVM/IVF/EC).

Histochemical detection of sugar residues in lizard teeth (Liolaemus gravenhorsti): a lectin-binding study.

The structural diversity of the many oligosaccharide chains of surface glycoconjugates renders them likely candidates for modulators of cell-interactions, cellular movements, differentiation, and cellular recognition. A selection of different lectins was used to investigate the appearance of cellular distribution and changes in sugar residues during tooth development in the polyphyodont lizard, Liolaemus gravenhorsti. Lectins from three groups were used: (1) N-acetylgalactosamine specificity: BS-1, PNA, RCA-120; (2) N-acetylglucosamine specificity: ECA; and (3) fucose specificity: UEA 1 and LTA.. Digital images were processed using Scion Image. Grayscale graphics in each image were obtained. The lectins used showed a strong, wide distribution of the L-fucose and N-acetylgalactosamine at the cell surface and in the cytoplasm of multinucleate odontoclast cell, while mononuclear odontoclast cells showed no binding, suggesting some roles that the residues sugar might play in the resorption of dentine or with multinucleation of odontoclast after the attachment to the dentine surface in this polyphyodont species. Further studies must be planned to determine the specific identities of these glycoconjugates,and to elucidate the roles played by these sugar residues in the complex processes related to odontogenesis in polyphyodont species.

Histochemical detection of sugar residues in lizard teeth (Liolaemus gravenhorsti): a lectin-binding study

The structural diversity of the many oligosaccharide chains of surface glycoconjugates renders them likely candidates for modulators of cell-interactions, cellular movements, differentiation, and cellular recognition. A selection of different lectins was used to investigate the appearance of cellular distribution and changes in sugar residues during tooth development in the polyphyodont lizard, Liolaemus gravenhorsti. Lectins from three groups were used: (1) N-acetylgalactosamine specificity: BS-1, PNA, RCA-120; (2) N-acetylglucosamine specificity: ECA; and (3) fucose specificity: UEA 1 and LTA.. Digital images were processed using Scion Image. Grayscale graphics in each image were obtained. The lectins used showed a strong, wide distribution of the L-fucose and N-acetylgalactosamine at the cell surface and in the cytoplasm of multinucleate odontoclast cell, while mononuclear odontoclast cells showed no binding, suggesting some roles that the residues sugar might play in the resorption of dentine or with multinucleation of odontoclast after the attachment to the dentine surface in this polyphyodont species. Further studies must be planned to determine the specific identities of these glycoconjugates,and to elucidate the roles played by these sugar residues in the complex processes related to odontogenesis in polyphyodont species.

Microscopic and histochemical study of odontoclasts in physiologic resorption of teeth of the polyphyodont lizard, Liolaemus gravenhorsti.

Using tartrate-resistant acid phosphatase (TRAP), we examined the cytodifferentiation of odontoclast cells in resorbing areas of dental tissues during the replacement of teeth in a polyphyodont lizard, Liolaemus gravenhorsti. We also report, by means of Lectin-HRP histochemistry, the distribution pattern of some specific sugar residues of TRAPase-positive cells. For detection of TRAPase activity, the azo dye-coupling technique was used. Lectin binding sites were demonstrated by means of specific HRP-lectins. The process of tooth resorption was divided into four stages: 1) preresorption-the wall of the dental pulp is covered with an odontoblast layer, and no TRAP-positive cells are in the dental pulp; 2) early resorption-TRAP-positive multinucleate odontoclasts are present on the dental wall, but the rest of the pulp surface is still covered with an odontoblast layer; 3) later resorption-the entire surface of the pulp chamber is lined with multinucleate odontoclasts; and 4) final resorption-the tooth has been totally resorbed. Odontoclasts are usually detached from the resorbed surface, and show signs of degeneration. Of the six lectins used, PNA, ECA, and UEA-1 bind to multinucleated but not mononuclear cells. All the remaining lectins, BS-1, RCA(120), and LTA showed no binding to any cells of the teeth. The significance of saccharidic moieties such as acetyl-galactosamine, acetyl-glucosamine, and fucose sugar residues is difficult to ascertain. Perhaps these oligosaccharides might be borne on molecules associated with odontoclastic resorption or associated with multinucleation of odontoclasts after attachment to the dentine surface.

Effect of cholinergic agonists on muscular tonus of the lizard small intestine and esophagus.

The underlying mechanisms of acetylcholine-induced intestinal relaxation in the lizard Liolaemus tenuis tenuis are still unknown. By using a classical model of intestinal recording of isometric contraction and relaxation in conjunction with specific pharmacological tools, this article studies the possible influence of EDRF/NO and nicotinic ganglionar receptors on the Ach-induced relaxation in an effort to elucidate the probable mechanisms involved in ACh effect. It was observed that the relaxation of the lizard intestine elicited by ACh (10(-7) - 4 x 10(-4) M) was not affected by hexametonium (5 x 10(-4) M) or tetrodotoxin (10(-6) M). Nicotine (10(-7) to 10(-4) M) induced relaxation was significantly antagonized by hexametonium; however, it was not influenced by tetrodotoxin. These results allow us to discard a neuronal pathway in cholinergic-induced relaxation, suggesting a more direct cholinergic effect on the smooth muscle, perhaps mediated by an unknown substance released by some specialized tissue. N-nitro-L-arginine, used to block NO-synthase and NO production, induced no changes in ACh-induced relaxation. Methylene blue, a soluble guanylate cyclase inhibitor, induced no changes in ACh-induced relaxation. These results allow us to discard a probable role of EDRF/nitric oxide in the ACh-induced relaxation of lizard small intestine, providing evidence that this mechanism could be different from that reported in other species.

Evaluación cintigráfica de patología nefrourológica diagnosticada en período antenatal / Scintigraphic assessment of nephrourologic pathology with prenatal diagnosis

Desde el advenimiento de la ecografía obstétrica, el diagnóstico antenatal (DA) de patología nefrourológica (PNU) es relativamente frecuente. El objetivo de este trabajo es evaluar retrospectivamente la cintigrafía renal dinámica (CRD) y estática (CRE) en 30 pacientes (pac) entre 2 días y 13 meses de vida, 19 de los 30 pacientes eran recién nacidos, de los cuales siete fueron estudiados durante la primera semana de vida. Se realizó CRD con Tc99n DTPA ó 13, CRE con Tc99m DMSA en 17 pacientes y ambos estudios en 14 pacientes. Trece de las 60 unidades (UR) (21.7 por ciento) estaban excluídas, uno de las cuales bilateral, secundaria a valvas de la uretra posterior. La causa más común de exclusión fue riñón multicístico. Siete de los doce pacientes con exclusión unilateral presentaron patología del riñon contralateral (EPU). En ninguno de estos siete pacientes existía compromiso significativo de la función renal al momento del cintigrama. En diez UR la CRD mostró un patrón obstructivo, siendo en la mayoría (6UR) secundaria a EPU. En 17 UR la CRD mostró retención pielocaliceal con respuesta normal al diurético, 14 UR eran hidronefrosis transitoria del recién nacido. En 11 niños se realizó seguimiento cintigráfico, siete postquirúrgico, en 5 de estos pacientes se observó una mejoría de la curva de excreción (71,4 por ciento). En tres de los 4 pacientes no operados el seguimiento cintigráfico demostró un deterioro en la función renal y/o en la curva de excreción, lo que motivó una corrección quirúrgica

Paro cardiorespiratorio: resultados de la reanimación en UTI pediátrica / Cardiorespiratory failure: results in pediatric ICU

Se estudian prospectivamente 71 pacientes que recibieron reanimación cardiopulmonar en una unidad de tratamiento intensivo pediátrico durante el período de un año. Aunque 39 pacientes (55%) se recuperaron inicialmente con las maniobras, 17 repitieron el paro cardíaco y fallecieron en las siguientes 24 horas. Sólo 12 elevado a 16,9 sobrevivían al alta del hospital. Ningún paciente con PCR de mas de 10 minutos de duración sobrevivió, a diferencia de 44% de reanimaciones exitosas en los casos de menor duración. La existencia de un evento desencadenante precisable, ausencia de apoyo ventilatorio mecánico preparo y enfermedad de origen respiratorio y no cardíaco, se asociaron con mejor pronóstico. La asistolía fue el tipo de actividad eléctrica terminal más frecuente (92%), ocurriendo sólo 4 casos de fibrilación ventricular en pacientes cardiópatas que no se recuperaron. Los pacientes que sobrevivieron no presentaron déficit neurológico y en un año de seguimiento sólo uno falleció a consecuencia de una malformación cardíaca congénita

Efecto crónico de Li en la presión arterial de rata / Chronic effect of Li ion on the blood pressure of the rat

El ion Li es absorbido con rapidez, se distribuye en todo el organismo y filtra en el glomérulo, pero es reabsorbido en un alto porcentaje por los túbulos. En el presente trabajo se investiga su papel fisiológico en la función renal, en relación con la presión arterial por ingestión crónica; para lo cual se observa presión arterial directa, volúmenes de orina, los electrolitos sodio y potasio y calicreína urinaria. A un grupo de ratas hembras de la cepa Wistar se les trató con solución de LiCl cada 48 horas por vía intragástrica y en forma crónica por un mes. Se realizaron observaciones cada diez días que demostraron los cambios que se producen en la presión arterial, los volúmenes y componentes de la orina: disminución de la, aumento del potasio plasmático, aumento de la calicreína urinaria y marcado efecto poliúrico durante todo el tratamiento, con marcado aumento de la excreción total de sodio y potasio urinarios