The vestibular calyceal junction is dismantled following subchronic streptomycin in rats and sensory epithelium stress in humans.

Hair cell (HC) loss by epithelial extrusion has been described to occur in the rodent vestibular system during chronic 3,3'-iminodipropionitrile (IDPN) ototoxicity. This is preceded by dismantlement of the calyceal junction in the contact between type I HC (HCI) and calyx afferent terminals. Here, we evaluated whether these phenomena have wider significance. First, we studied rats receiving seven different doses of streptomycin, ranging from 100 to 800 mg/kg/day, for 3-8 weeks. Streptomycin caused loss of vestibular function associated with partial loss of HCI and decreased expression of contactin-associated protein (CASPR1), denoting calyceal junction dismantlement, in the calyces encasing the surviving HCI. Additional molecular and ultrastructural data supported the conclusion that HC-calyx detachment precede HCI loss by extrusion. Animals allowed to survive after the treatment showed functional recuperation and rebuilding of the calyceal junction. Second, we evaluated human sensory epithelia obtained during therapeutic labyrinthectomies and trans-labyrinthine tumour excisions. Some samples showed abnormal CASPR1 label strongly suggestive of calyceal junction dismantlement. Therefore, reversible dismantlement of the vestibular calyceal junction may be a common response triggered by chronic stress, including ototoxic stress, before HCI loss. This may partly explain clinical observations of reversion in function loss after aminoglycoside exposure.

Pre- and intra-operative prognostic factors of facial nerve function in cerebellopontine angle surgery.

PURPOSE: The study assesses whether pre- and intraoperative factors linked to electromyography and direct electrical stimulation (DES) of facial nerve can predict facial nerve function in the short- (12 days) and long-term (1 year) after cerebellopontine angle (CPA) tumor resection. METHODS: 157 patients who underwent surgical resection of CPA tumors with facial nerve monitoring. Pre-operative factors (age, tumor size, pure tone average), surgical time and intra-operative parameters regarding facial function, minimum stimulation threshold (MST), compound muscle action potential (CMAP) and the difference between proximal and distal CMAP (DPDC) were evaluated. RESULTS: A correlation between tumor size, MST, CMAP and facial function in both short and long term was found. A higher grade of immediate facial paralysis corresponded to a higher risk of poor outcome after one year. A postoperative House-Brackmann (HB) score of V or VI was correlated with poor outcome in 88.8% and 93.8% of cases. A risk of HB 3 or more, in the long term, was correlated with a tumor size of 20.2 mm. Using an MST of 0.1 mA, for long-term predictions, sensitivity and specificity were 0.62 (95% CI 0.46-0.75) and 0.73 (95% CI 0.61-0.82), respectively. With a CMAP cut-off < 200 µV, for long-term prediction, sensitivity was 0.73 (95% CI 0.53-0.87) and specificity 0.73 (95% CI 0.55-0.85). CONCLUSION: The assessment based on the cut-offs described increases the ability to predict facial function. Improving predictive accuracy enables surgeons to address patients' expectations and to establish an intervention timeline for planning facial reanimation.

Validación de un cuestionario para medir conocimientos sobre medicamentos en adolescentes españoles / To validate a medication knowledge questionnaire in the adolescent high school student population

Objetivo: validar un cuestionario de conocimiento de medicación en la población adolescente estudiante de bachillerato.Material y métodos: emplazamiento y muestra: para la citada validación se han usado los datos obtenidos en el estudio ConóceMe. Concretamente, la respuesta al cuestionario en el primer seguimiento en el total de los alumnos (n= 12.030). Para la prueba de fiabilidad test-retest se han usado la línea base y el primer seguimiento solo del grupo control (n= 6.228).Análisis de los datos: se han evaluado la fiabilidad (consistencia interna, test de dos mitades y test-retest), la validez (análisis factorial) y el funcionamiento de los ítems (dificultad, discriminación y correlación respecto al total del cuestionario). Resultados: las pruebas de fiabilidad arrojan unos resultados con índices todos ellos superiores a 0,7. Así el alfa de Cronbach tiene un valor de 0,80 y el test de dos mitades 0,81.La validez muestra un análisis factorial que obtiene su óptimo en una solución de tres factores, todos ellos con valores propios superiores a 0,9 y explicando entre los tres una variancia del 70 % del total.En cuanto a los ítems, todos presentan valores aceptables de funcionamiento en cuanto a dificultad y discriminación y una buena correlación respecto al total de puntuación del cuestionario.Conclusiones: el cuestionario de conocimiento valorado en el presente estudio presenta unas propiedades métricas adecuadas, siendo de utilidad para el fin que se pretendía, evaluar conocimientos generales sobre medicamentos en la población de estudiantes de bachillerato.

Actualización del papel del farmacéutico comunitario en el manejo del paciente asmático / Update on the role of the community pharmacist in the management of the asthmatic patient

El asma es una de las enfermedades crónicas no transmisibles más frecuentes a nivel mundial, siendo además la más prevalente de las enfermedades crónicas respiratorias. En España se estima que la prevalenciageneral del asma en niños es de un 10% y de cerca del 5% en edad adulta. Los pacientes afectados poresta enfermedad presentan una peor calidad de vida, menor productividad laboral y más comorbilidadesasociadas, y a pesar de las mejoras acontecidas en las últimas décadas en el manejo y tratamiento de estapatología, y a que se considera al asma como una enfermedad manejable y tratable, todavía se siguenproduciendo muertes por ataques de asma. Llegados a este punto, es donde aparece la figura del farmacéutico comunitario que atendiendo a su formación y al particular emplazamiento donde desarrolla sulabor asistencial, puede y debe jugar un papel decisivo en el abordaje del paciente asmático en atenciónprimaria con el fin de colaborar activamente con otros profesionales sanitarios para ayudar a revertir lasituación actual. Ese abordaje podría realizarse a través de diferentes intervenciones a desarrollar dentrodel servicio integral de atención al paciente asmático, como son: el cribado de pacientes asmáticos nodiagnosticados, la detección de situaciones de riesgo y de pacientes con asma poco o mal controlada,la detección de pacientes en situación de sobreuso o abuso de medicación de rescate, la detección depacientes con baja adhesión al tratamiento antiinflamatorio con corticoides inhalados, la revisión del usode los dispositivos de inhalación, la educación sanitaria y la ayuda en la obtención de planes de acciónpersonalizados por escrito. (AU)

Asthma is one of the most common chronic non-communicable diseases worldwide, being also the mostprevalent chronic respiratory disease. In Spain, the overall prevalence of asthma in children is estimatedto be 10% and around 5% in adults. Patients affected by this disease have a poorer quality of life, lowerwork productivity and more associated comorbidities, and despite the improvements that have occurredin recent decades in the management and treatment of this pathology, and despite the fact that asthma isconsidered a manageable and treatable disease, deaths from asthma attacks are still occurring. It is at thispoint that the figure of the community pharmacist comes into play. Given their training and the particularlocation where they develop their activity, they can and should play a decisive role in the approach toasthma patients in primary care in order to actively collaborate with other healthcare professionals tohelp to reverse the current situation. This approach could be carried out through different interventionswithin the comprehensive asthmatic patient care service, such as; screening of undiagnosed asthmaticpatients, detection of risk situations and patients with poorly or poorly controlled asthma, detection ofpatients in a situation of over-reliance in rescue medication, detection of patients with low adherence toanti-inflammatory treatment with inhaled corticosteroids, review of the use of inhalation devices, healtheducation, and help in obtaining personalised written action plans. (AU)

CT Assessment of Otic Capsule Bone Density in Paget's Disease of the Temporal Bone and Its Relationship With Hearing Loss.

OBJECTIVES: To study computed tomography findings in Paget's disease of temporal bone (PDTB) and analyze the relations between otic capsule bone mineral density values measured in Hounsfield Units (HU) and hearing loss (HL). STUDY DESIGN: Observational case-control study. SETTING: Tertiary referral center. PATIENTS: Radiographically confirmed PDTB cases and control group. INTERVENTION: Diagnostic. MAIN OUTCOME MEASURES: Hearing thresholds and computed tomography bone density values. RESULTS: Twenty-three ears in the case group (PDTB) and 27 control ears were included. In the PDTB group, HL was found in 87% of the ears (43% mixed) and an air-conduction threshold of 50.7 dB (SD = 19.8). In the control group, 48% of the ears showed HL (7% mixed) and an air-conduction threshold of 34.5 dB (SD = 20.6) was found; the difference was statistically significant (p < 0.05). Measurements of bone density (HU) in the otic capsule (regions of interest [ROI] 1 and 2) and in the petrous bone (ROI 3) were significantly lower (p < 0.05) in the PDTB group than in controls.The PDTB group presented a significant association between otic capsule bone density in ROI 1 and mean otic capsule density with air and bone-conduction thresholds (p < 0.05). In controls, no association was observed between any density value and audiometric thresholds. CONCLUSION: PDTB patients showed more frequent HL, lower thresholds, and a higher proportion of mixed HL than controls. Bone density (HU) was decreased in all ROIs in PDTB patients in comparison with controls. Bone density in the otic capsule was associated with HL in PDTB patients, but no association was observed between bone density and HL in controls.

Diagnostic Accuracy of Intracochlear Test Electrode for Acoustic Nerve Monitoring in Vestibular Schwannoma Surgery.

OBJECTIVES: Cochlear implants (CIs) are a well-known hearing restoration option for patients with vestibular schwannoma (VS) in cases of neurofibromatosis type-2 and, more recently, for patients with sporadic VS. One of the main limitations when performing CI during VS surgery is the capability to preserve the acoustic nerve (AN) anatomically and functionally. Significant efforts have been directed toward developing an intraoperative testing method for monitoring the AN function to determine if, after tumor removal, it is suitable for conducting stimuli delivered by a CI. However, all these methods have significant limitations, and none of them have documented diagnostic efficacy. To overcome these limitations and to obtain reliable information before CI insertion, a minimally invasive intracochlear test electrode (TE) has been recently developed. This TE has demonstrated to be suitable to test the integrity of the AN before CI in patients without any residual hearing by recording electrically evoked auditory brainstem responses (EABR). The present study constitutes the next phase of this research, which was to determine the usefulness of EABR obtained intraoperatively with the intracochlear TE after the resection of a VS and to calculate its diagnostic accuracy to assess the functionality of the AN for CI. DESIGN: This was a prospective, multicenter study of diagnostic accuracy. It was conducted in three tertiary referral centers between January 2015 and 2018. This study was designed following the Standards for Reporting of Diagnostic Accuracy Studies (STARD) statement guidelines. The STARD statement are guidelines to improve the completeness and transparency of reports of diagnostic accuracy studies. The diagnostic accuracy of the EABR evoked with the intracochlear TE after tumor removal was studied. Accuracy, sensitivity, specificity, positive predictive value, and negative predictive value were calculated. Patients eligible for the study were consecutive adults undergoing surgery for VS with simultaneous CI. The test under evaluation (index test) was the EABR obtained with the intracochlear TE after resection of the tumor. The reference test (gold standard) was the presence of auditory perception with the CI, defined as the presence of sound detection on an audiogram at 500, 1000, 2000, and 4000 Hz of no greater than 50 dB. In all the cases, auditory perception was verified by the presence of a positive EABR evoked with the CI. RESULTS: Twenty-one patients were included during the study period; seven patients were excluded from the diagnostic efficacy analysis due to inconclusive EABR results or absence of the gold standard to compare (they did not finally receive the CI). Thus, the outcome of the gold standard was assessed in 14 cases: 9 cases had positive EABR, all of them obtained auditory perception with the CI, and 5 cases had negative EABR, only one case had auditory perception with the CI, which constitutes the only false negative of this study. Accuracy of the TE was 93% (95% confidence interval, 66 to 100%), sensitivity 90% (95% confidence interval, 71 to 100%), specificity 100% (95% confidence interval, 100 to 100%), positive predictive value 100% (95% confidence interval, 100 to 100%), and negative predictive value 80% (95% confidence interval, 45 to 100%). CONCLUSIONS: EABR elicited with the intracochlear TE had a diagnostic accuracy of 93% for predicting auditory perception with CIs after VS removal. These results suggest that the intracochlear TE can be used intraoperatively after tumor removal to test the integrity of the AN as a useful tool to complement the surgeon's perception for decision-making regarding implantation.

Conciliación y revisión de la medicación al alta en paciente sometido a gastrostomía para alimentación enteral / Reconciliation and review of medication at discharge in a patient undergoing gastrostomy for enteral feeding

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Mitochondrial DNA and TLR9 drive muscle inflammation upon Opa1 deficiency.

Opa1 participates in inner mitochondrial membrane fusion and cristae morphogenesis. Here, we show that muscle-specific Opa1 ablation causes reduced muscle fiber size, dysfunctional mitochondria, enhanced Fgf21, and muscle inflammation characterized by NF-κB activation, and enhanced expression of pro-inflammatory genes. Chronic sodium salicylate treatment ameliorated muscle alterations and reduced the muscle expression of Fgf21. Muscle inflammation was an early event during the progression of the disease and occurred before macrophage infiltration, indicating that it is a primary response to Opa1 deficiency. Moreover, Opa1 repression in muscle cells also resulted in NF-κB activation and inflammation in the absence of necrosis and/or apoptosis, thereby revealing that the activation is a cell-autonomous process and independent of cell death. The effects of Opa1 deficiency on the expression NF-κB target genes and inflammation were absent upon mitochondrial DNA depletion. Under Opa1 deficiency, blockage or repression of TLR9 prevented NF-κB activation and inflammation. Taken together, our results reveal that Opa1 deficiency in muscle causes initial mitochondrial alterations that lead to TLR9 activation, and inflammation, which contributes to enhanced Fgf21 expression and to growth impairment.

Promoción de los servicios profesionales farmacéuticos. "El farmacéutico que necesitas": carpas de salud 2015-2016 / Promotion of professional pharmaceutical services. "The pharmacist you need": health tents 2015-2016

Introducción: La Sociedad Española de Farmacia Familiar y Comunitaria (SEFAC), en colaboración con el laboratorio farmacéutico Ratiopharm, ha llevado a cabo el proyecto de carpas de servicios profesionales farmacéuticos El farmacéutico que necesitas en 2015 y 2016. Objetivos: Dar a conocer a la sociedad y los pacientes las actividades del farmacéutico comunitario en la prestación de servicios profesionales farmacéuticos (SPF) y valorar su satisfacción con la actividad. Material y métodos: El proyecto se basa en el ofrecimiento a la población general de actuaciones de SPF adaptados al ámbito de una estructura itinerante, desmontable, tipo carpa. Los servicios ofrecidos fueron: uso adecuado del medicamento, cesación tabáquica, cribado de enfermedad pulmonar obstructiva crónica, consejo nutricional general, cribado de malnutrición en mayores de 65 años, cribado de diabetes, medida de la presión arterial y cálculo del riesgo vascular. Los SPF fueron realizados por farmacéuticos comunitarios (FC) voluntarios locales, previamente formados con protocolos de trabajo diseñados por SEFAC para su actuación en este entorno. Resultados: El evento se realizó en 15 ciudades diferentes, participaron 655 FC voluntarios, atendiendo a un total de 2.531 usuarios a los que se realizaron 7.290 actuaciones profesionales, con una media de 3 por usuario. La mayoría de los usuarios, encuestados tras recibir los servicios, declararon haber mejorado el conocimiento sobre su estado de salud y lo que desde la farmacia comunitaria se les puede ofrecer. Conclusiones: Un elevado número de usuarios participantes manifestaron haber incrementado sus conocimientos sobre los SPF y su estado de salud y la práctica totalidad valoró de forma positiva las actividades realizadas (AU)

Introduction: The Spanish Society for Family and Community Pharmacy (SEFAC in Spanish), in collaboration with the Ratiopharm pharmaceutical laboratory, carried out the project of professional pharmaceutical service tents, The Pharmacist You Need, in 2015 and 2016. Objectives: Raise the awareness of society and patients of the activities of community pharmacists in the provision of professional pharmaceutical services (PPS) and evaluate their satisfaction with the activity. Materials and Methods: The project is based on offering to the general public PPS actions adapted to the format of a mobile, temporary, tent-type structure. The services offered were: proper use of medicines, smoking cessation, screening for chronic obstructive pulmonary disease, general nutritional advice, screening for malnutrition in people over 65, screening for diabetes, blood pressure measurement and calculation of vascular risk. The PPS were delivered by volunteer local community pharmacists (CP) who had received prior training on work protocols designed by SEFAC for its action in this setting. Results: The event was held in 15 different cities, 655 volunteer CPs participated, attending to a total of 2,531 users who received 7,290 professional actions, an average of three per user. Most of the users surveyed after receiving the services said that their awareness of their health status and what a community pharmacy can offer them had improved. Conclusions: A high number of the users that participated stated that their awareness of PPS and their health status had improved, and practically all of them rated the actions performed positively (AU)

Early Genetic Diagnosis of Neurofibromatosis Type 2 From Skin Plaque Plexiform Schwannomas in Childhood.

Importance: Neurofibromatosis type 2 (NF2) is a devastating genetic condition characterized by the development of multiple tumors of the nervous system. An early diagnosis of individuals with NF2 would facilitate treatment and reduction of disease impact because most severe effects of the disease do not usually develop before adolescence. Little attention has traditionally been paid to dermatological signs in NF2. However, skin plaques are commonly seen in patients with NF2, normally appearing either at birth or early childhood, providing an opportunity for early NF2 detection and testing. Objective: To determine the clinical utility of skin plaque identification and characterization in children for reaching an early diagnosis of patients with NF2 and to evaluate their molecular pathogenesis and their use in the genetic diagnostics of NF2. Design, Setting, and Participants: Diagnostic test study by the histological and genetic characterization of skin plaques from patients with NF2. Patients were 7 individuals with NF2 or clinical suspicion of NF2 treated at the Spanish Reference Center on Phakomatoses. Main Outcomes and Measures: Histological evaluation of all skin plaques was performed. Fresh skin plaques were cultured to obtain Schwann cells and the NF2 gene was genetically analyzed. For all 7 patients, NF2 clinical history was reviewed. Results: In all 7 patients (4 male and 3 female), all skin plaques analyzed were histologically characterized as plexiform schwannomas. Genetic analysis of primary Schwann cell cultures derived from them allowed the identification of a constitutional and a somatic NF2 mutation. Genetic testing allowed the early diagnosis of NF2 in a child only exhibiting the presence of skin plaques. Most of the patients with NF2 analyzed had an early presentation of skin plaques and a severe NF2 phenotype. Conclusions and Relevance: This work emphasizes the clinical utility of a careful dermatological inspection and the correct identification of skin plaques in children for an early diagnosis of NF2. We show for the first time that Schwann cells derived from skin plaque plexiform schwannomas bear the double inactivation of the NF2 gene and thus constitute an excellent source of tissue for genetic testing, especially in the context of mosaicism.

Consulta farmacéutica por somnolencia en un lactante / Pharmaceutical enquiry by drowsiness in breastfed baby

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Lactate induces FGF21 expression in adipocytes through a p38-MAPK pathway.

FGF21 (fibroblast growth factor 21), first described as a main fasting-responsive molecule in the liver, has been shown to act as a true metabolic regulator in additional tissues, including muscle and adipose tissues. In the present study, we found that the expression and secretion of FGF21 was very rapidly increased following lactate exposure in adipocytes. Using different pharmacological and knockout mice models, we demonstrated that lactate regulates Fgf21 expression through a NADH/NAD-independent pathway, but requires active p38-MAPK (mitogen activated protein kinase) signalling. We also demonstrated that this effect is not restricted to lactate as additional metabolites including pyruvate and ketone bodies also activated the FGF21 stress response. FGF21 release by adipose cells in response to an excess of intermediate metabolites may represent a physiological mechanism by which the sensing of environmental metabolic conditions results in the release of FGF21 to improve metabolic adaptations.

Detección precoz de EPOC en las farmacias comunitarias de Baleares (FARBALEPOC) / Early detection of COPD in community pharmacies in the Balearic Islands (FARBALEPOC)

Introducción: Importantes estudios como el EPISCAN ha puesto de manifiesto en España, el retraso diagnóstico de la EPOC y una elevada tasa de infradiagnóstico, encontrando que el 73% de los pacientes estaba sin diagnosticar y en la mayoría de los que se diagnosticaban ya se encontraban en estadios III y IV. Este retraso en el diagnóstico provoca una elevada morbilidad, así como un elevado coste asistencial. Por todo esto es necesario un diagnóstico precoz. Objetivos: Identificar pacientes de EPOC no diagnosticados, mediante un servicio de cribado de EPOC ofrecido por farmacias comunitarias. Material y métodos: Población de estudio: mayores de 40 años de ambos sexos, fumadores, exfumadores, y fumadores pasivos, que presentan signos o síntomas crónicos, y que en el cuestionario COPD-PS obtengan una puntuación ≥4. A los pacientes se les realizó la prueba de espiración forzada mediante el dispositivo portátil Vitalograph COPD-6. Si presentaba valores del cociente FEV1/FEV6 <0,75 se derivaba el paciente a su médico de familia para que se le realizase la espirometría convencional de confirmación. Resultados: Se encuestaron un total de 198 pacientes, 154 de ellos presentaban COPD-PS ≥4,38 (24,7%) de ellos presentaron resultados del cociente FEV1/FEV6 <0,75. La prevalencia de posible EPOC en este estudio se ha estimado en un 19,2% de los pacientes encuestados. Conclusiones: El significativo porcentaje de pacientes a los se les ha detectado una posible EPOC, nos demuestra la utilidad de implantar servicios de cribado y detección precoz de EPOC en las farmacias comunitarias (AU)

Introduction: Important studies, as EPISCAN, have revealed in Spain a delay in diagnosis of COPD and a high rate of underdiagnosis, finding that 73% of patients were undiagnosed and most of those who were already diagnosed were in III and IV stages of COPD. This delay in diagnosis causes high morbidity and high health care costs. For all that, early diagnosis is necessary. Objectives: To identify undiagnosed COPD patients, using a COPD screening service offered by community pharmacies. Material and methods: Study Population: current, former and passive smokers, aged 40 or older who have signs or chronic symptoms were recruited. Those who obtained a score ≥ 4 in COPD-PS questionnaire underwent a forced expiratory test by using Vitalograph COPD-6 portable device. If the FEV1 / FEV6 value was <0.75, the patient was referred to the general practitioner (GP) in order to confirm COPD diagnosis with conventional spirometry. Results: A total of 198 patients were recruited, 154 of them had a COPD-PS score ≥ 4, of whom 38 (24.67%) presented results of FEV1 /FEV6 <0.75. The prevalence of COPD in this study may have been estimated at 19.2% of patients enrolled. Discussion and conclusions: The significant proportion of patients detected with a possible diagnosis of COPD demonstrates the utility to implement services of screening and early detection of COPD in community pharmacies (AU)

Segmental neurofibromatosis type 2: discriminating two hit from four hit in a patient presenting multiple schwannomas confined to one limb.

BACKGROUND: A clinical overlap exists between mosaic Neurofibromatosis Type 2 and sporadic Schwannomatosis conditions. In these cases a molecular analysis of tumors is recommended for a proper genetic diagnostics. This analysis is challenged by the fact that schwannomas in both conditions bear a somatic double inactivation of the NF2 gene. However, SMARCB1-associated schwannomas follow a four-hit, three-step model, in which both alleles of SMARCB1 and NF2 genes are inactivated in the tumor, with one of the steps being always the loss of a big part of chromosome 22 involving both loci. CASE PRESENTATION: Here we report a 36-year-old woman who only presented multiple subcutaneous schwannomas on her right leg. To help discriminate between both possible diagnoses, an exhaustive molecular genetic and genomic analysis was performed on two schwannomas of the patient, consisting in cDNA and DNA sequencing, MLPA, microsatellite multiplex PCR and SNP-array analyses. The loss of a big part of chromosome 22 (22q12.1q13.33) was identified in both tumors. However, this loss involved the NF2 but not the SMARCB1 locus. SNP-array analysis revealed the presence of the same deletion breakpoint in both schwannomas, indicating that this alteration was actually the first NF2 inactivating hit. In addition, a distinct NF2 point mutation in each tumor was identified, representing independent second hits. In accordance with these results, no deletions or point mutations in the SMARCB1 gene were identified. None of the mutations were present in the blood. Two of the patient's children inherited chromosome 22 deleted in schwannomas of the mother, but in its wild type form. CONCLUSIONS: These results conclusively confirm the segmental mosaic NF2 nature of the clinical phenotype presented.

Fibroblast growth factor 21 protects the heart from oxidative stress.

AIMS: Oxidative stress mediated by reactive oxygen species (ROS) plays a striking role in the pathogenesis of heart failure, and antioxidants have been shown to attenuate cardiac remodelling in experimental models of cardiac damage. We recently showed that fibroblast growth factor 21 (Fgf21) is produced by the heart and exerts protective effects, preventing cardiac hypertrophy development. The aim of the study was to determine the effects of Fgf21 during oxidative stress signalling in the heart. METHODS AND RESULTS: Fgf21 treatment in cardiomyocytes in culture induced the expression of genes encoding proteins involved in antioxidative pathways, including mitochondrial uncoupling proteins (Ucp2 and Ucp3) and superoxide dismutase-2 (Sod2) and reduced ROS production. In keeping with this, expression of antioxidant genes in response to lipopolysaccharide (LPS)-induced stimulation of pro-inflammatory pathways or isoproterenol-induced cardiac hypertrophy in the heart was reduced in Fgf21-null mice. Moreover, we found that Fgf21 is expressed in and released by cardiomyocytes in response to LPS, and its expression is under the control of the Sirt1 (sirtuin-1) pathway. This Fgf21 released by cardiomyocytes acts in an autocrine manner to protect cells against oxidative stress. Finally, failing human hearts showed up-regulation of Fgf21, Ucp3, and Sod2, confirming the association between Fgf21 induction and the control of cardiac oxidative stress pathways. CONCLUSION: Our data indicate that Fgf21 regulates genes involved in antioxidant pathways in an autocrine manner, thus preventing ROS production in cardiac cells. Therefore, Fgf21 acts as an antioxidant factor in the heart, preventing induction of pro-oxidative pathways by inflammatory or hypertrophic conditions.

FGF21 expression and release in muscle cells: involvement of MyoD and regulation by mitochondria-driven signalling.

Although the liver is generally considered the main site of production of FGF21 (fibroblast growth factor-21), high FGF21 levels have been found to be associated with neuromuscular mitochondrial genetic diseases, and there are indications that the muscle may be a relevant site of FGF21 production under conditions of muscular mitochondrial stress. In the present study, we found that expression and release of FGF21 was associated with myogenic differentiation, and we identified MyoD as a major controller of FGF21 gene transcription. Mimicking mitochondrial dysfunction using respiratory chain/oxidative phosphorylation inhibitors resulted in enhanced expression and release of FGF21 by muscle cells. The increased production of reactive oxygen species, subsequent induction of p38 MAPK (mitogen-activated protein kinase) and activation of an ATF2 (activating transcription factor 2)-binding site at the proximal promoter region of the FGF21 gene was found to be a major mechanism linking mitochondrial dysfunction with enhanced FGF21 gene transcription in myogenic cells. The myogenic factor MyoD was required for the induction of FGF21 gene transcription by mitochondrial dysfunction, thus explaining the preferential response of muscle cells to mitochondrial dysfunction-induced FGF21 expression and secretion. FGF21 release by muscle cells in response to mitochondrial alterations may represent a physiological mechanism by which the sensing of internal energetic status by muscles results in the release of FGF21 to favour systemic metabolic adaptations.

FGF21 mediates the lipid metabolism response to amino acid starvation.

Lipogenic gene expression in liver is repressed in mice upon leucine deprivation. The hormone fibroblast growth factor 21 (FGF21), which is critical to the adaptive metabolic response to starvation, is also induced under amino acid deprivation. Upon leucine deprivation, we found that FGF21 is needed to repress expression of lipogenic genes in liver and white adipose tissue, and stimulate phosphorylation of hormone-sensitive lipase in white adipose tissue. The increased expression of Ucp1 in brown adipose tissue under these circumstances is also impaired in FGF21-deficient mice. Our results demonstrate the important role of FGF21 in the regulation of lipid metabolism during amino acid starvation.

In vitro antisense therapeutics for a deep intronic mutation causing Neurofibromatosis type 2.

Neurofibromatosis type 2 (NF2) is an autosomal-dominant disorder affecting about 1:33 000 newborns, mainly characterized by the development of tumors of the nervous system and ocular abnormalities. Around 85% of germline NF2 mutations are point mutations. Among them, ∼25% affect splicing and are associated with a variable disease severity. In the context of our NF2 Multidisciplinary Clinics, we have identified a patient fulfilling clinical criteria for the disease and exhibiting a severe phenotype. The patient carries a deep intronic mutation (g. 74409T>A, NG_009057.1) that produces the insertion of a cryptic exon of 167pb in the mature mRNA between exons 13 and 14, resulting in a truncated merlin protein (p.Pro482Profs*39). A mutation-specific antisense phosphorodiamidate morpholino oligomer was designed and used in vitro to effectively restore normal NF2 splicing in patient-derived primary fibroblasts. In addition, merlin protein levels were greatly recovered after morpholino treatment, decreasing patient's fibroblasts in vitro proliferation capacity and restoring cytoeskeleton organization. To our knowledge, this is the first NF2 case caused by a deep intronic mutation in which an in vitro antisense therapeutic approximation has been tested. These results open the possibility of using this approach in vivo for this type of mutation causing NF2.

Peroxisome proliferator-activated receptor-gamma coactivator-1alpha controls transcription of the Sirt3 gene, an essential component of the thermogenic brown adipocyte phenotype.

Sirt3 (silent mating type information regulation 2, homolog 3), a member of the sirtuin family of protein deacetylases with multiple actions on metabolism and gene expression is expressed in association with brown adipocyte differentiation. Using Sirt3-null brown adipocytes, we determined that Sirt3 is required for an appropriate responsiveness of cells to noradrenergic, cAMP-mediated activation of the expression of brown adipose tissue thermogenic genes. The transcriptional coactivator Pgc-1α (peroxisome proliferator-activated receptor-γ coactivator-1α) induced Sirt3 gene expression in white adipocytes and embryonic fibroblasts as part of its overall induction of a brown adipose tissue-specific pattern of gene expression. In cells lacking Sirt3, Pgc-1α failed to fully induce the expression of brown fat-specific thermogenic genes. Pgc-1α activates Sirt3 gene transcription through coactivation of the orphan nuclear receptor Err (estrogen-related receptor)-α, which bound the proximal Sirt3 gene promoter region. Errα knockdown assays indicated that Errα is required for full induction of Sirt3 gene expression in response to Pgc-1α. The present results indicate that Pgc-1α controls Sirt3 gene expression and this action is an essential component of the overall mechanisms by which Pgc-1α induces the full acquisition of a brown adipocyte differentiated phenotype.

Neurofibromatosis tipo 2 / Neurofibromatosis type 2

La neurofibromatosis tipo 2 es una enfermedad invalidante que se hereda de forma autosómica dominante. A menudo se ha confundido con la neurofibromatosis tipo 1, aunque son patologías distintas. Todos los sujetos que hereden una mutación en el gen de la neurofibromatosis tipo 2 (NF2), desarrollarán dicha enfermedad, caracterizada por el crecimiento de schwanomas, habitualmente vestibulares y de forma bilateral, así como meningiomas u otros tumores benignos del sistema nervioso central, antes de los 30 años de edad. Actualmente, podemos identificar la mutación del NF2 en la mayoría de las familias afectas. Hasta un 20% de los pacientes afectos de NF2 sin historia familiar, aparentemente casos esporádicos, son en realidad individuos con mosaicismo para esa mutación. La morbilidad de esos tumores es en gran medida debida a su tratamiento, que es principalmente quirúrgico. Cuando son pequeños, los schwanomas vestibulares se pueden resecar completamente con preservación tanto de la función auditiva como facial. En caso de tumores grandes se puede colocar un implante coclear o bien de tronco cerebral durante el mismo acto quirúrgico. Los principales factores pronósticos son: la edad media al diagnóstico, la presencia de meningiomas intracraneales y si el paciente fue tratado o no en un centro especializado (AU)

Type 2 neurofibromatosis (NF2) is an invalidating, inherited, dominant, autosomal disease. It is commonly confused with type 1 neurofibromatosis, although the two disorders are different. All subjects who inherit a mutated NF2 gene will develop the disease, which is characterised by the growth of schwannomas, generally affecting the vestibular nerve bilaterally, as well as meningiomas and other benign central nervous system tumours, before their third decade of life. It is currently possible to identify the NF2 mutation in most affected families. Up to about 20% of NF2 patients with no family history, apparently sporadic cases, are actually individuals with mosaicism for this mutation. Much of the morbidity from these tumours results from their treatment, which is primarily surgical. Small vestibular schwannomas can often be completely resected with preservation of both hearing and facial function. In case of large tumours it is possible to place a cochlear or brain stem implant during the schwannoma surgery. Age at diagnosis, the presence of intracranial meningiomas, and whether the patient was treated at a specialty centre or not, have been cited as the strongest prognostic factors (AU)