Grupo de seguridad del paciente del colegio oficial de farmacéuticos: 10 años de práctica colaborativa entre farmacias y centros sanitarios / Patient safety group of the official college of pharmacists: 10 years of collaborative practice between pharmacies and healthcare centres

JUSTIFICACIÓN: la colaboración entre centros sanitarios y farmacias es esencial para mejorar la seguridad de los pacientes y el estado de salud de la población. El Grupo de Seguridad del Paciente del Colegio Oficial de Farmacéuticos se constituyó en 2011 para fomentar la comunicación entre niveles asistenciales y mejorar la atención a los pacientes.OBJETIVOS: describir las actividades del Grupo, enfocadas a la práctica colaborativa entre farmacias y centros del Área Sanitaria del Servicio de Salud, para mejorar la seguridad de los pacientes.MATERIAL Y MÉTODOS: el Grupo lo integran profesionales de la farmacia comunitaria, hospitalaria, de atención primaria y del Centro de Información de Medicamentos colegial, que se reúnen mensualmente para elaborar documentos, promover acciones formativas y fomentar la mejora de la comunicación entre las farmacias y los centros sanitarios. También se revisan incidentes en la prescripción o dispensación de medicamentos, para evitar errores de seguridad en el ámbito extrahospitalario. Los materiales elaborados se difunden entre los colegiados, otros profesionales sanitarios y/o la población general.RESULTADOS/DISCUSIÓN: las principales actividades desarrolladas por el Grupo de Seguridad del Paciente han sido: -Elaboración de un protocolo de comunicación entre farmacias y centros sanitarios.-Difusión e implantación de un formulario de notificación de incidentes, para su posterior evaluación por el Grupo de Seguridad.-Revisión de incidencias relacionadas con errores de prescripción, errores de dispensación y problemas de funcionamiento de receta electrónica, que pueden dar lugar a errores de medicación. (AU)

Plasma Aß42/40 Ratio Detects Early Stages of Alzheimer's Disease and Correlates with CSF and Neuroimaging Biomarkers in the AB255 Study.

BACKGROUND: Easily accessible biomarkers are needed for the early identification of individuals at risk of developing Alzheimer's disease (AD) in large population screening strategies. OBJECTIVES: This study evaluated the potential of plasma ß-amyloid (Aß) biomarkers in identifying early stages of AD and predicting cognitive decline over the following two years. DESIGN: Total plasma Aß42/40 ratio (TP42/40) was determined in 83 cognitively normal individuals (CN) and 145 subjects with amnestic mild cognitive impairment (a-MCI) stratified by an FDG-PET AD-risk pattern. RESULTS: Significant lower TP42/40 ratio was found in a-MCI patients compared to CN. Moreover, a-MCIs with a high-risk FDG-PET pattern for AD showed even lower plasma ratio levels. Low TP42/40 at baseline increased the risk of progression to dementia by 70%. Furthermore, TP42/40 was inversely associated with neocortical amyloid deposition (measured with PiB-PET) and was concordant with the AD biomarker profile in cerebrospinal fluid (CSF). CONCLUSIONS: TP42/40 demonstrated value in the identification of individuals suffering a-MCI, in the prediction of progression to dementia, and in the detection of underlying AD pathology revealed by FDG-PET, Amyloid-PET and CSF biomarkers, being, thus, consistently associated with all the well-established indicators of AD.

¿En qué hemos cambiado a lo largo de una década? Estudio comparativo de la asistencia sanitaria prestada en Líbano (Operación UNIFIL) en 2006 y 2016 / What have we changed over a decade? Comparative study of healthcare provided in Lebanon (UNIFIL Operation) in 2006 and 2016

Introducción: En 2016, España cumple una década de presencia en Líbano dentro de la operación de Naciones Unidas (UNIFIL) United Nations Interim Force in Lebanon. El objetivo del presente estudio es analizar la evolución de la asistencia sanitaria prestada en dos contingentes españoles desplegados en esa zona de operaciones con una diferencia de 10 años. Material y métodos: Estudio transversal descriptivo retrospectivo realizado durante dos periodos (del 8 septiembre al 8 de noviembre de 2006 y del 3 de septiembre al 18 de noviembre de 2016). La población a estudio fueron los pacientes atendidos en el primer escalón sanitario durante los citados periodos. Resultados: Los datos del primer y segundo intervalo respectivamente fueron: efectivos desplegados 523/562; asistencias médicas realizadas 1168/1435; enfermedades digestivas 269/423; enfermedades otorrinolaringológicas 222/147; cirugía menor 187/75; enfermedad dermatológica 175/128; traumatología 152/413; enfermedad odontológica 58/128; enfermedad oftalmológica 53/16; otras 70/105; evacuaciones a ROLE 2 4/76 y evacuaciones a ROLE 4 0/0. Conclusión: Hay similitud entre las atenciones sanitarias realizadas en ambos periodos de tiempo, excepto en las enfermedades traumatológicas donde aparece un incremento de casi un 300% en el segundo intervalo de tiempo respecto al primero

Introduction: In 2016, Spain celebrates a decade of presence in Lebanon within the United Nations Interim Force in Lebanon (UNIFIL). The aim of the present study is to analyse the evolution of the health care provided in two Spanish contingents deployed in that area of operations, comparing two periods within ten years of elapsed time. Material and method: A cross-sectional retrospective study was carried out during two periods (from the 8th of September to the 8th of November of 2006 and from the 3rd of September to the18th of November of 2016). The study population was the patients treated in the first Medical Treatment Facility during those periods. Results: The data of the first and second time interval respectively were: deployed troops 523/562; medical assistance performed 1168/1435; Digestive disease 269/423; Otorhinolaryngological diseases 222/147; Minor surgery 187/75; Dermatological disease 175/128; Traumatology 152/413; Dental disease 58/128; Opthalmologic disease 53/16; Other 70/105; Evacuations to ROLE 2 4/76 and evacuations to ROLE 4 0/0. Conclusion: There is a similarity between the healthcare provided in both periods of time, except in the traumatic diseases, because there was an increase of almost 300% during the second interval of time (AU)

Ultrastructure of Lingual Papillae in Common Chimpanzee (Pan troglodytes) Foetus, Newborn and Adult Specimens.

Among primates, the two recognized species of chimpanzees (common chimpanzee, Pan troglodytes; pygmy chimpanzee, Pan paniscus) are considered to be the most similar to humans. Importantly, in mammals, the food intake behaviour largely determines the tongue morphology, including the type, proportion and distribution of gustatory and non-gustatory tongue papillae. The lingual papillae form during its development and mature in post-natal life depending on the different feeding. In this study, we have used scanning electron microscopy to analyse the age-related changes in the lingual papillae of foetal, newborn and adult P. troglodytes. Four main types of lingual papillae, denominated filiform, fungiform, foliate and vallate, and one subtype of filiform papillae called conical papillae, were found. The main age-related changes observed in all kinds of papillae were a progressive keratinization and morphological complexity along the lifespan. During the foetal period, there was scarce keratinization, which progressively increases in young animals to adulthood. The number of filiform increased with ageing, and both filiform and fungiform papillae in adult tongues are divided into pseudopapillae. On the other hand, the vallate papillae vary from smooth simple surfaces in foetal tongues to irregular surfaces with grooves and pseudopapillae (microscopic papilla-shaped formations within the papilla itself) in adults. These results describe for the first time the age-related variations in the three-dimensional aspect of lingual papillae of the chimpanzee tongue and provide new data to characterize more precisely these structures in the human closest specie.

Evaluación clínica de la mascarilla laríngea Baska Mask ® en pacientes adultos en cirugía mayor ambulatoria / Clinical evaluation of the Baska Mask ® laryngeal mask in adult patients in ambulatory surgery

Objetivo. Valorar el comportamiento clínico de la Baska Mask®, un nuevo dispositivo supraglótico de ventilación que incorpora un balón autoinflable y doble canal de aspiración. Material y métodos. Se incluyeron prospectivamente 80 pacientes sin criterios de vía aérea difícil. Se evaluaron el número de intentos y la facilidad de inserción, la ventilación, la presión de sellado de la vía aérea, la visión fibrobroncoscópica, el acceso gástrico y las complicaciones observadas. Se analizaron y compararon los tamaños 3, 4 y 5. Resultados. La tasa de éxito de inserción al primer intento fue del 88%, y la global, del 100%, aunque en el 44% de los casos fue necesario rotar o curvar el dispositivo. La ventilación fue eficaz en el 96%, requiriendo maniobras de ajuste en el 39%. La presión de sellado fue de 33 ± 7 cmH2O. El tamaño 3 requirió significativamente menos ajustes y obtuvo una presión de sellado más alta. La visión total o parcial de las cuerdas vocales se obtuvo en el 90% de los 66 casos evaluados, en el 5% se observó obstrucción parcial por distorsión del borde libre del balón, y en el 5% restante no se identificaron estructuras glóticas. La inserción de la sonda gástrica fue fácil en todos los casos. Las complicaciones fueron leves y transitorias. Conclusiones. La Baska Mask consiguió una presión de sellado elevada, una ventilación adecuada y un acceso gástrico rápido. Sin embargo, requirió con frecuencia aplicar maniobras de ajuste para la inserción y para obtener una ventilación adecuada (AU)

Objective. To evaluate the clinical performance of the Baska Mask®, a new second-generation supraglottic airway device with a self-inflating cuff and two side suction channels for continuous aspiration. Material and methods. Eighty adult patients without difficult airways were prospectively included. Ease of insertion and number of attempts needed, quality of ventilation, airway seal pressure, fibreoptic view, ease of gastric access, and complications were assessed. Sizes 3, 4, 5 were analyzed and compared. Results. First attempt insertion success rate was 88% and the overall rate was 100%, although additional maneuvers were necessary in 44% of the cases. The ventilation was adequate in 96%, with 39% of them requiring adjusting maneuvers. Size 3 needed significantly less adjustments, and achieved a higher seal pressure than sizes 4 and 5 combined. The airway seal pressure was 33 ± 7 cmH2O. Complete or partial vocal cords were visible in 90% of the 66 cases assessed. Partial obstruction, caused by distortion of the cuff-free border, was seen in 5%, and no glottic structures were identified in 5%. Gastric access was easy in all cases. Complications were mild and transient. Conclusions. The Baska Mask achieves a high seal pressure, effective ventilation, and a quick access to drain gastric contents. However, additional adjustment maneuvers are frequently required to insert the mask and to optimize ventilation (AU)

Clinical evaluation of the Baska Mask laryngeal mask in adult patients in ambulatory surgery.

OBJECTIVE: To evaluate the clinical performance of the Baska Mask, a new second-generation supraglottic airway device with a self-inflating cuff and two side suction channels for continuous aspiration. MATERIAL AND METHODS: Eighty adult patients without difficult airways were prospectively included. Ease of insertion and number of attempts needed, quality of ventilation, airway seal pressure, fibreoptic view, ease of gastric access, and complications were assessed. Sizes 3, 4, 5 were analyzed and compared. RESULTS: First attempt insertion success rate was 88% and the overall rate was 100%, although additional maneuvers were necessary in 44% of the cases. The ventilation was adequate in 96%, with 39% of them requiring adjusting maneuvers. Size 3 needed significantly less adjustments, and achieved a higher seal pressure than sizes 4 and 5 combined. The airway seal pressure was 33 ± 7 cm H2O. Complete or partial vocal cords were visible in 90% of the 66 cases assessed. Partial obstruction, caused by distortion of the cuff-free border, was seen in 5%, and no glottic structures were identified in 5%. Gastric access was easy in all cases. Complications were mild and transient. CONCLUSIONS: The Baska Mask achieves a high seal pressure, effective ventilation, and a quick access to drain gastric contents. However, additional adjustment maneuvers are frequently required to insert the mask and to optimize ventilation.

Brain-derived neurotrofic factor and its receptor TrkB are present, but segregated, within mature cutaneous Pacinian corpuscles of Macaca fascicularis.

Some mechanoreceptors in mammals depend totally or in part on the neurotrophins brain-derived neurotrophic factor (BDNF) and neurotrophin-4 (NT-4), and their receptor TrkB, for development and maintenance. These actions are presumably exerced regulating the survival of discrete sensory neurons in the dorsal root ganglia which form mechanoreceptors at the periphery. In addition, the cells forming the mechanoreceptors also express both neurotrophins and their receptors although large differences have been described among species. Pacinian corpuscles are rapidly adapting low-threshold mechanoreceptors whose dependence from neurotrophins is not known. In the present study, we analyzed expression of TrkB and their ligands BDNF and NT-4 in the cutaneous Pacinian corpuscles of Macaca fascicularis using immunohistochemistry and fluorescent microscopy. TrkB immunoreactivity was found in Pacinian corpuscles where it co-localized with neuron-specific enolase, and occasionally with S100 protein, thus suggesting that TrkB expression is primarily into axons but also in the lamellar cells and even in the outer core. On the other hand, BDNF immunoreactivity was found the inner core cells where it co-localized with S100 protein but also in the innermost layers of the outer core; NT-4 immunostaining was not detected. These results describe for the first time the expression and distribution of a full neurotrophin system in the axon-inner core complex of mature Pacinian corpuscles. The data support previous findings demonstrating large differences in the expression of BDNF-TrkB in mammalian mechanoreceptors, and also suggest the existence of a retrograde trophic signaling mechanism to maintain morphological and functional integrity of sensory neurons supplying Pacinian corpuscles.

Immunohistochemical detection of the putative mechanoproteins ASIC2 and TRPV4 in avian herbst sensory corpuscles.

The avian Herbst corpuscles are the equivalent of the Pacinian corpuscles in mammals, and detect vibration and the movement of joints and feathers. Therefore, they can be regarded as rapidly adapting low-threshold mechanoreceptors. In recent years, it has been establish that some ion channels are involved in mechanosensation and are present in both mechanosensory neurons and mechanoreceptors. Here we have used immunohistochemistry to localize some putative mechanoproteins in the Herbst corpuscles from the rictus of Columba livia. The proteins investigated were the subunits of the epithelial Na(+) channel (ENaC), the transient-receptor potential vanilloid 4 (TRPV4), and the acid-sensing ion channel 2 (ASIC2). Immunoreactivity for ENaC subunits was never found in Herbst corpuscles, while the axon expressed ASIC2 and TRPV4 immunoreactivity. Moreover, TRPV4 was also detected in the cell forming the inner core. The present results demonstrate for the first time the occurrence of mechanoproteins in avian low-threshold mechanoreceptors and provide further evidence for a possible role of the ion channels in mechanosensation.

Immunohistochemical localization of acid-sensing ion channel 2 (ASIC2) in cutaneous Meissner and Pacinian corpuscles of Macaca fascicularis.

Acid-sensing ion channel 2 (ASIC2) is a member of the degenerin/epithelial sodium channel superfamily, presumably involved mechanosensation. Expression of ASIC2 has been detected in mechanosensory neurons as well as in both axons and Schwann-like cells of cutaneous mechanoreceptors. In these studies we analysed expression of ASIC2 in the cutaneous sensory corpuscles of Macaca fascicularis using immunohistochemistry and laser confocal-scanner microscopy. ASIC2 immunoreactivity was detected in both Meissner and Pacinian corpuscles. It was found to co-localize with neuron-specific enolase and RT-97, but not with S100 protein, demonstrating that ASIC2 expression is restricted to axons supplying mechanoreceptors. These results demonstrate for the first time the presence of the protein ASIC2 in cutaneous rapidly adapting low-threshold mechanoreceptors of monkey, suggesting a role of this ion channel in touch sense.

Reduction of glial fibrillary acidic protein-immunoreactive astrocytes in some brain areas of old hairless rhino-j mice (hr-rh-j).

Mutations in the hairless (hr) gene of mice result in hair follicle and other epithelial defects. The hr gene is expressed at high levels in the brain where it probably participates in the survival and maintenance of some neuronal populations, but whether it also supports glial populations of the central nervous system has been not investigated. To clarify this, quantitative immunohistochemistry for astrocytes (glial fibrillary acidic protein (GFAP)) and microglial cells (CD11b macrophage antigen) was used in the brain of a mutant mouse strain, the hairless (hr-rh-j) type, which carries the homozygous hr gene rhino mutation. The glial cell density was assessed in the cerebral cortex, hippocampus, striatum, hypothalamus and cerebellum of young (3 months) and old (9 months) hr-rh-j mice. No significant differences were found between young wild-type and hr-rh-j mice. The density of GFAP immunoreactive astrocytes normally increased as a function of age, but in older hr-rh-j mice there was a severe reduction (P<0.01) in the striatum, hypothalamus, and hippocampus. Conversely, the microglial cells were insensible to aging or to hr-rh-j mutation. These results suggest that the hr gene is involved in the maintenance of the GFAP immunoreactive cells in some cerebral areas. Nevertheless, because these animals do not show any neurological signs, the functional significance of the present findings remains to be established.

Seudoaneurismas infecciosos en pacientes drogadictos: ligadura o revascularización / Infectious false aneurysm in drug addicts: ligation or revascularization?

Introducción. La punción arterial en drogadictos puede producir falsos aneurismas infecciosos (FAI) con riesgo de hemorragia, pérdida de la extremidad o muerte. Pacientes y métodos. Siete pacientes drogadictos con FAI fueron tratados en nuestro hospital entre 1990 y 1999. Una masa pulsátil con fistulización de líquido serosanguinolento fue la presentación característica. El estudio arteriográfico distinguió entre absceso y seudoaneurisma, lo que ayudó a la localización anatómica de las lesiones. Una reconstrucción arterial, electiva (n = 3) o urgente (n = 4), se practicó en todos los casos. Resultados. El cultivo microbiano fue siempre positivo. Se utilizó un injerto venoso en 6 casos y se requirió una prótesis de politetrafluoroetileno (PTFE) en un paciente con ausencia de venas útiles. Aunque el postoperatorio inmediato transcurrió con normalidad, la recidiva de infección y la hemorragia masiva obligaron a la ligadura arterial urgente en todos los pacientes. Tan sólo fue necesaria una amputación de brazo. No se registraron muertes. Conclusiones. La reconstrucción arterial en pacientes drogadictos se acompaña de numerosas complicaciones. La ligadura arterial con resección amplia de los tejidos es segura, bien tolerada y con aceptable morbilidad (AU)

The thymus of the hairless rhino-j (hr/rh-j) mice.

The hairless (hr) gene is expressed in a large number of tissues, primarily the skin, and a mutation in the hr gene is responsible for the typical cutaneous phenotype of hairless mice. Mutant hr mouse strains show immune defects involving especially T cells and macrophages, as well as an age-related immunodeficiency and an accelerated atrophy of the thymus. These data suggest that the hr mutation causes a defect of this organ, although hr transcripts have not been detected in fetal or adult mice thymus. The present study analyses the thymus of young (3 mo) and adult (9 mo) homozygous hr-rh-j mice (a strain of hairless mice) by means of structural techniques and immunohistochemistry to selectively identify thymic epithelial cells, dendritic cells, and macrophages. There were structural alterations in the thymus of both young and adult rh-rh-j mice, which were more severe in older animals. These alterations consisted of relative cortical atrophy, enlargement of blood vessels, proliferation of perivascular connective tissue, and the appearance of cysts. hr-rh-j mice also showed a decrease in the number of epithelial and dendritic cells, and macrophages. Taken together, present results strongly suggest degeneration and accelerated age-dependent regression of the thymus in hr-rh-j mice, which could explain at least in part the immune defects reported in hairless mouse strains.

Intermediate filament proteins in developing and adult human dorsal root and sympathetic ganglia / Las proteínas de los filamentos intermedios en los ganglios simpáticos y de la raíz dorsal humana adulta en desarrollo

Neuronal maturation in the central nervous system, as well as in some cells deriving from neural crest, is accompanied by a swich in the expression of cytoskeletal intermediate filament proteins. Whether this occurs in humans and the exact timing of this change in human dorsal root and sympathetic ganglia are matters still open to debate. The present study was designed to analyze these issues in human embryos (estimated gestational age -e.g.a.- ranging between 6 and 12 weeks), as well as the possible co-expression of more than one intermediate filament protein in both embryos and adults. A panel of commercially available antibodies against vimentin, glial fibrillary acidic protein and neurofilament proteins was used. Glial fibrillary acidic protein was consistently absent in both developing and adult dorsal root or sympathetic ganglia. Conversely, embryonic neurons, satellite glial cells and Schwann cells displayed vimentin immunoreactivity. The number of vimentin immunoreactive neurons decreased progressively, and it was absent from neurons by 12 weeks e.g.a., while it persisted in satellite glial and Schwann cells. By adulthood, the pattern of distribution was identical. The occurrence of neurofilament proteins in peripheral neurons was a regular feature from early developmental stages to adulthood, and a time-dependent increase in the percentage of neurons containing phosphorylated neurofilaments was observed. The present results demonstrate that developing human dorsal root and sympathetic ganglion neurons co-express vimentin and neurofilaments for a short time, but that the intermediate filaments for mature neurons are neurofilaments. Our findings also show that co-expression or a switch in the expression of intermediate filament proteins do not occur in satellite glial cells or Schwann cells, which normally contain vimentin and not glial fibrillary acidic protein (AU)

La maduración neuronal en poblaciones neuronales discretas del sistema central nervioso, así como en algunas células que derivan de la cresta neural es acompañada de un cambio en la expresión de las proteínas citoesqueléticas de filamentos intermedios. El que esto ocurre en los seres humanos y la cronología exacta de este cambio en los ganglios de la raíz dorsal y simpáticos no se ha dilucidado todavía. El presente estudio fue diseñado para analizar estos problemas en embriones humanos (edad gestacional estimada –e.g.e- en el intervalo entre 6 y 12 semanas), así como la posible co-expresión de más de una proteína de filamentos intermedios, tanto en embriones como en adultos. Se utilizó un panel de anticuerpos comerciales frente a la vimentina, a la proteína fibrilar glial ácida, y a las proteínas de los neurofilamentos. La proteína fibrilar glial ácida estaba consistentemente ausente de los ganglios de la raíz dorsal y simpáticos, tanto en el adulto como en embriones en desarrollo. Por el contrario, las neuronas embrionarias, las células gliales satélites y las células de Schwann exhibían inmunoreactividad frente a la vimentina. El número de neuronas con inmunoreactividad para la vimentina descendió progresivamente y este compuesto no se detectaba en las neuronas a una e.g.e. de 12 semanas, mientras que persistía en las células Schwann. Llegada la edad adulta, el patrón de distribución era idéntico. La presencia de proteínas de neurofilamentos en las neuronas periféricas se apreció como una característica consistente a partir de las fases tempranas del desarrollo hasta la edad adulta, observándose un descenso dependiente del tiempo en el porcentaje de neuronas que contenían neurofilamentos fosforilados. Los resultados aquí obtenidos demuestran que las neuronas de los ganglios de la raíz dorsal y simpáticos humanos en desarrollo co-expresan la vimentina y los neurofilamentos durante un periodo corto de tiempo, pero los filamentos intermedios de las neuronas maduras son neurofilamentos. Nuestros hallazgos también muestran que la co-expresión o un cambio en la expresión de las proteínas de los filamentos intermedios no ocurren en las células gliales ni en las de Schwann, las cuales normalmente contienen vimentina y no la proteína fibrilar glial ácida (AU)

Insulin-like growth factor 1-receptor (IGF-1R) in developing and adult human dorsal root ganglia / El factor de crecimiento tipo insulina receptor-1 (IGF-1R) en los ganglios de la raíz dorsal humana en desarrollo y adulta

Insulin-like growth factors (IGFs) promote neurite outgrowth in cultured sensory neurons that binding to IGF type 1 receptor (IGF-1R) and seem to be involved in the pathogenesis of some metabolic and toxic peripheral neuropathies coursing with altered sensitivity. However, the distribution of IGF-1R in the human sensory peripheral nervous system is unknown. In this study, we used light immunohistochemistry to analyse the occurrence and localization of IGF-1R in developing (6 to 22 weeks of estimated gestational age, e.g.a.) and adult (age range 25-41 years) human dorsal root ganglia (DRG). In human embryos (6-8 weeks e.g.a.) and young foetuses (9 weeks e.g.a.), most neurons (84%, 92%, and 83%, respectively) displayed IGF-1R immunoreactivity (IR). In older foetuses (12 and 22 weeks e.g.a.), the number of immunoreactive neurons decreased progressively (78 and 68%, respectively) reaching values similar to those observed in adults (64%). The subpopulation of adult primary sensory neurons showing IGF-1R IR covered the entire size range, but the neurons were mainly small. Furthermore, in adults all satellite glial cells and Schwann cells were immunoreactive. The present results suggest a role for IGF-1R in the differentiation and maturation of primary sensory neurons, and in the maintenance of a subset of them in adulthood, as well as in the control of peripheral glial cells (Schwann and satellite glial cells). These findings might serve as a basis for future studies in pathologic DRG, in which IGF-1R or its ligands may be involved (AU)

Los factores de crecimiento tipo insulina (IGFs) promueven el crecimiento de neuritas en neuronas sensitivas en cultivo que se unen al IGF receptor tipo 1 (IGF-1R) y parecen estar implicados en la patogénesis de algunas neuropatías periféricas metabólicas y tóxicas cursando con alteraciones de la sensibilidad. Sin embargo, se desconoce la distribución de IGF-1R en el sistema nervioso periférico sensitivo humano. En este estudio, usamos inmunohistoquímica a microscopía óptica para analizar la existencia y localización de IGF-1R en los ganglios de la raíz dorsal humana (DRG) en desarrollo (6 a 22 semanas de edad estimada de gestación, e.g.a.) y adulta (rango de edad de 25-41 años). En embriones humanos (6-8 semanas e.g.a.) y fetos jóvenes (9 semanas e.g.a.), la mayoría de las neuronas (84 por ciento, 92 por ciento, y 83 por ciento, respectivamente) mostraron inmunorreactividad (IR) a IGF-1R. En fetos de más edad (12 y 22 semanas e.g.a.), el número de neuronas inmunorreactivas disminuyó progresivamente (78 y 68 por ciento, respectivamente), alcanzando valores similares a los observados en los adultos (64 por ciento). La subpoblación de neuronas sensitivas primarias adultas que mostraron IGF-1R IR cubrieron un rango completo de tamaños, pero las neuronas fueron principalmente pequeñas. Además, en adultos todas las células satélites gliales y las células de Schwann fueron inmunorreactivas. Los presentes resultados sugieren un papel para IGF-1R en la diferenciación y maduración de las neuronas sensitivas primarias, y en el mantenimiento de una subpoblación de ellas en el estado adulto, así como en el control de las células gliales periféricas (Schwann y células satélites gliales). Estos hallazgos podrían servir como base para futuros estudios de los ganglios de la raíz dorsal patológicos, en los que el IGF-1R o sus ligandos pueden estar implicados (AU)

Trks and p75 genes are differentially expressed in the inner ear of human embryos. What may Trks and p75 null mutant mice suggest on human development?

Recent work has shown the expression of Neurotrophins low (p75) and high affinity (Trk's A, B, and C) receptors in the developing inner ear sensory neurons of chick and mouse. Likewise the biological significance of such receptor expression was demonstrated by using both Trks and Neurotrophins null mutant mice. The present study was conducted to determine the expression of Trks and p75 proteins in the human inner ear throughout development. Hence to assess the potential role of Neurotrophins in the development of auditory and vestibular specific innervation in man. In other words, we intend to address the issue whether or not what null mutant mice for Trks and p75 have revealed on inner ear development may be relevant for human embryos. Fifty-two inner ears and their cochleovestibular ganglions (CVG) from human embryos and fetuses, ranging from 5 to 24 weeks of pregnancy were analyzed. Both Western blot and immunocytochemistry on frozen sections were used as complementary procedures. Quantitative Western blot studies revealed that Trk-B and C immunoreactivity (IR) appeared by embryonic week 5 in CVG neurons, increased at high levels between embryonic weeks 7 and 12, and later on, in 15 week-old specimens and older began to decrease to minimal levels. Trk-A IR was detected at just moderate levels during 5 and 7 weeks reflecting the presence of NGF high affinity receptors only at these earlier developmental ages. The p75 IR was detected at high degrees in the early stage of the 5th week and at abundant levels in all studied inner ears from the 7th to the 24th pregnancy week. These Western blot observations were corroborated by immunocytochemistry on frozen sections, which also revealed a major distribution of both p75 and Trks on neuronal bodies while p75 appears localized on supporting cells. Our findings reveal a tight correlation between p75 and Trks expression throughout human development and specific inner ear developmental events, such as target-dependent neuronal cell death and afferent hair cells innervation. That kind of association of p75 and Trks temporal pattern with distinctive steps in inner ear developmental schedule, is a feature shared between human embryos and other mammals, such as mouse. Based on the present results and considering them together with the reported phenotype of p75 and Trks null mutant mice, we hypothesize that p75 and Trk receptors, as well as, their binding Neurotrophins may be essential in human inner ear development. Accordingly, they may be required molecules for sensory epitheliums innervation and target-dependent neuronal cell death, during embryogenesis and even early postnatal life, in man.

The hairless gene of the mouse: relationship of phenotypic effects with expression profile and genotype.

Various mutations of the hairless (hr) gene of mice result in hair loss and other integument defects. To examine the role of the hr gene in mouse development, the expression profile of hr has been determined by in situ hybridisation and correlated to the nature of genetic changes and morphological abnormalities in different mutant animals. Four variant alleles have been characterised at the molecular level. hr/hr mice produce reduced, but significant, levels of hr mRNA whereas other alleles contain mutations which would be expected to preclude the synthesis of functional product, demonstrating a correlation between allelic variation at the hr locus and phenotypic severity. hr expression was shown to be widespread and temporally regulated. It was identified in novel tissues such as cartilage, developing tooth, inner ear, retina, and colon as well as in skin and brain. Analysis of mice homozygous for the rhino allele of hairless revealed that, although no morphological defects were detectable in many tissues normally expressing hr, previously undescribed abnormalities were present in several tissues including inner ear, retina, and colon. These findings indicate that the hairless gene product plays a wider role in development than previously suspected. Dev Dyn 1999;216:113-126.

Changes in the cerebellar cortex of hairless Rhino-J mice (hr-rh-j).

A mutation in the hr gene is responsible for typical epithelium phenotype in hairless mice. As this gene is expressed at high levels not only in the skin but also in the brain, the aim of the study was to clarify its role in the central nervous system. We have analyzed by morphological and immunocytochemical methods (calbindin D-28k, phosphorylated and 200 kDa neurofilament protein) the cerebellum of a mutated mouse strain, the hairless (hr-rh-j) type carrying the homozygous hr gene rhino mutation. The cerebellar cortex was studied in young (3 months) and adult (9 months) wild type and mutated mice. No major structural change was found in any of the groups and neuronal density or neuronal arrangement were similar in mutated animals to their age-matched controls. Nevertheless there were changes in shape and size of the Purkinje neurons in the old mutated animals respect to their normal littermates, while the molecular and the granule cell layers were apparently invariable. Calbindin (CB) immunohistochemistry revealed a significant decrease in the expression of this protein in the Purkinje cells of the aged mutated mice. Immunohistochemistry for a neurofilament protein (NFP) showed a reduction of staining in all the cerebellar cortex layers in the older animals, which was much more evident in the (hr-rh-j) mutated mice. These results suggest that hr gene is involved in the structural maintenance of the mature cerebellar cortex, rather than in the development. Our findings may also be consistent with an accelerated aging of the central nervous system in rh-rh-j mice.

Differential expression of microtubule associated protein MAP-2 in developing cochleovestibular neurons and its modulation by neurotrophin-3.

Microtubule associated proteins (MAPs) are essential cytoskeletal proteins in developing neurons. The present study was undertaken to analyze the expression of MAP2 and its isoforms (a,b,c) during the embryonal and early post-hatching development of chicken cochleovestibular ganglion (CVG) neurons. Moreover, we have investigated MAP2 expression in primary cultures of CVG neurons, and whether it is regulated by neurotrophin-3 (NT3). The expression of MAP2 immunoreactivity (IR) was studied using both Western blot and immunohistochemistry on tissue sections and primary cultures. In vivo MAP2c was expressed from incubation day 4 (E4) to E10, and MAP2b was found in all embryonal stages studied and at post-hatching day 10 (P10), whereas MAP2a was restricted to the post-hatching periods. The cellular localization of IR was in the neuronal perikarya and their peripheral processes (dendrites) but not in axons. Primary cultures matched the in vivo pattern of MAP2 expression, and IR was localized in neuronal cell bodies and the initial segment of the neuronal processes. Exogenous NT3 regulated the expression of MAP2 isoforms in a dose dependent manner. At the survival dose of 0.5 ng/ml NT3, the main MAP2 expression was MAP2c. Conversely, at the neuritogenic dose of 5 ng/ml NT3 increased MAP2b and MAP2a expression, but not MAP2c. The present results demonstrate that MAP2 isoforms are developmentally regulated, thus suggesting that each isoform is specifically involved in CVG neuron maturation. Furthermore, we provide evidence of MAP2 regulation in culture by the neurotrophic factor NT3.

Expression of the cytoskeletal protein MAP5 and its regulation by neurotrophin 3 (NT3) in the inner ear sensory neurons.

Microtubule-associated proteins (MAPs) are essential cytoskeletal components during development for neurogenesis and neuronal plasticity. Inner ear innervation is accomplished by cochleovestibular ganglion (CVG) neurons in a highly specific, well-defined pattern, which is regulated by neurotrophic factors belonging to the neurotrophin family. The inner ear offers a suitable model for studying the expression of MAPs and assessing their role in neurotrophin-induced effects that are required for neuron-target innervation. The present study was undertaken to analyze the expression and localization of MAP5 isoforms during development of CVG neurons in vivo an in vitro; as well as the regulation of MAP5 by neurotrophin-3 (NT3) in cell culture. MAP5 expression in the inner ear of chick embryos and postnatal specimens was monitored using immunoblots and immunohistochemistry on frozen sections. MAP5 was highly expressed during the early stages of CVG development, at embryonic day (E)4, being located in both neuronal perikarya and neurites. Expression was maintained during the neurite outgrowth phase (E6-E12), when strong MAP5 immunostaining was observed at the same cellular locations. MAP5 expression decreased suddenly at E14, after the establishment of specific connections between the CVG neurons and their targets, the sensory epithelium of the inner ear. In cultured CVG neurons addition of NT3 led to increased MAP5 expression and produced neurite outgrowth. Both effects are differentially regulated in parallel by low (0.5 ng/ml) and high (5 ng/ml) NT3 concentrations. Present results suggest that MAP5 may be involved in neurotrophin-induced microtubule bundling during neurite outgrowth of auditory neurons.