Morphometric Predictors of Penile Length Increase After Division of its Suspensory Ligament.

INTRODUCTION: Division of the suspensory ligament of the penis has emerged as a popular surgical approach for penile lengthening, but accurate preoperative predictions of lengthening outcomes remain elusive. This study aimed to identify readily measurable anatomical parameters associated with post-ligamentolysis penile length gain, facilitating more reliable preoperative estimations. METHODS: An experimental cross-sectional study was performed on 16 adult cadavers. Data collected before dissection included: age at death, ethnicity, height, length of the penis before dissection and width of the suspensory ligament of penis. Following the complete dissection of the suspensory ligament of penis, the depth of the pubic symphysis and the penile length after the procedure were measured. The absolute and relative length differences pre- and post-ligamentolysis were calculated. Correlation coefficients were used to study relations between these variables. RESULTS: Penile length increased uniformly after complete division of the suspensory ligament (average gain: 26.38 mm, SD = 14.83 mm; range 4-60 mm). Pearson correlation revealed a significant negative correlation between pre-ligamentolysis penile length and post-ligamentolysis increase (r = - 0.601; p = 0.014), suggesting greater gains in individuals with shorter pre-ligamentolysis lengths. Age, ligament width, and pubic arch depth showed no significant correlations. Ethnicity did not impact post-ligamentolysis length increase (t = - 0.135; p = 0.894). CONCLUSIONS: This study highlights the potential to predict penile length gain post-ligamentolysis through measurable anatomical parameters. The ability to anticipate the outcome of this procedure could empower surgeons to provide informed counseling, potentially elevating patient satisfaction. An experimental cross-sectional study was performed to investigate the outcomes of penile lengthening surgery Penile lengthening was achieved in all subjects via complete dissection of the suspensory ligament of the penis Penile length increase may be predicted preoperatively using easily measurable anatomical parameters NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

Workload associated with manual assessment of vital signs as compared with continuous wireless monitoring.

BACKGROUND: Vital sign monitoring is considered an essential aspect of clinical care in hospitals. In general wards, this relies on intermittent manual assessments performed by clinical staff at intervals of up to 12 h. In recent years, continuous monitoring of vital signs has been introduced to the clinic, with improved patient outcomes being one of several potential benefits. The aim of this study was to determine the workload difference between continuous monitoring and manual monitoring of vital signs as part of the National Early Warning Score (NEWS). METHODS: Three wireless sensors continuously monitored blood pressure, heart rate, respiratory rate, and peripheral oxygen saturation in 20 patients admitted to the general hospital ward. The duration needed for equipment set-up and maintenance for continuous monitoring in a 24-h period was recorded and compared with the time spent on manual assessments and documentation of vital signs performed by clinical staff according to the NEWS. RESULTS: The time used for continuous monitoring was 6.0 (IQR 3.2; 7.2) min per patient per day vs. 14 (9.7; 32) min per patient per day for the NEWS. Median difference in duration for monitoring of vital signs was 9.9 (95% CI 5.6; 21) min per patient per day between NEWS and continuous monitoring (p < .001). Time used for continuous monitoring in isolated patients was 6.6 (4.6; 12) min per patient per day as compared with 22 (9.7; 94) min per patient per day for NEWS. CONCLUSION: The use of continuous monitoring was associated with a significant reduction in workload in terms of time for monitoring as compared with manual assessment of vital signs.

Distinct brain-wide presynaptic networks underlie the functional identity of individual cortical neurons.

Neuronal connections provide the scaffolding for neuronal function. Revealing the connectivity of functionally identified individual neurons is necessary to understand how activity patterns emerge and support behavior. Yet, the brain-wide presynaptic wiring rules that lay the foundation for the functional selectivity of individual neurons remain largely unexplored. Cortical neurons, even in primary sensory cortex, are heterogeneous in their selectivity, not only to sensory stimuli but also to multiple aspects of behavior. Here, to investigate presynaptic connectivity rules underlying the selectivity of pyramidal neurons to behavioral state 1-12 in primary somatosensory cortex (S1), we used two-photon calcium imaging, neuropharmacology, single-cell based monosynaptic input tracing, and optogenetics. We show that behavioral state-dependent neuronal activity patterns are stable over time. These are not determined by neuromodulatory inputs but are instead driven by glutamatergic inputs. Analysis of brain-wide presynaptic networks of individual neurons with distinct behavioral state-dependent activity profiles revealed characteristic patterns of anatomical input. While both behavioral state-related and unrelated neurons had a similar pattern of local inputs within S1, their long-range glutamatergic inputs differed. Individual cortical neurons, irrespective of their functional properties, received converging inputs from the main S1-projecting areas. Yet, neurons that tracked behavioral state received a smaller proportion of motor cortical inputs and a larger proportion of thalamic inputs. Optogenetic suppression of thalamic inputs reduced behavioral state-dependent activity in S1, but this activity was not externally driven. Our results revealed distinct long-range glutamatergic inputs as a substrate for preconfigured network dynamics associated with behavioral state.

Incidência e evitabilidade de eventos adversos no pronto atendimento: estudo retrospectivo / Incidencia y evitabilidad de eventos adversos en unidad de pronta atención: estudio retrospectivo / Incidence and avoidability of adverse events in emergency care: a retrospective study

Resumo Objetivo Analisar a incidência e a evitabilidade de eventos adversos em pacientes adultos internados em uma unidade de pronto atendimento no município do interior mineiro. Métodos Estudo de coorte, com revisão retrospectiva de 296 prontuários por meio do uso de formulários de rastreamento (fase 1) e avaliação (fase 2), preenchidos por profissionais não médicos e médico, respectivamente. Na primeira fase foram avaliados os dados demográficos, condição clínica dos pacientes, potenciais eventos adversos, e a verificação da qualidade das informações registradas nos prontuários. Na segunda fase, identificou-se e caracterizou-se os eventos adversos previamente rastreados. Resultados Observou-se a predominância de pessoas do sexo masculino, da raça branca, com 60 anos ou mais. A incidência de eventos adversos acumulada foi de 4,4% (n=13), sendo todos considerados evitáveis, e caracterizados como de gravidade leve, em sua maioria. De acordo com a natureza, 81,2% dos eventos adversos estavam relacionados ao cuidado em geral. Já em relação aos fatores intrínsecos e extrínsecos, encontrou-se uma média de 1,17 e 1,5 por paciente, respectivamente. Constatou-se que 17 pacientes foram identificados com potencial evento adverso, com 22 incidentes na análise. Conclusão Identificou-se que 100% dos eventos adversos eram evitáveis e a maioria relacionados ao cuidado em geral. Esses achados reforçam a necessidade de se avaliar e compreender a incidência e a evitabilidade dos eventos adversos em unidades de pronto atendimento, o que permite o planejamento e implementação de estratégias que visem o desenvolvimento de ações de controle e resposta a estes eventos, garantindo um atendimento seguro e de qualidade.

Resumen Objetivo Analizar la incidencia y la evitabilidad de eventos adversos en pacientes adultos internados en una unidad de pronta atención en un municipio del interior del estado de Minas Gerais. Métodos Estudio de cohorte, con revisión retrospectiva de 296 historias clínicas mediante el uso de formularios de rastreo (fase 1) y evaluación (fase 2), completados por profesionales no médicos y médicos, respectivamente. En la primera fase se evaluaron los datos demográficos, la condición clínica de los pacientes, los potenciales eventos adversos y la verificación de la calidad de la información registrada en las historias clínicas. En la segunda fase, se identificaron y caracterizaron los eventos adversos previamente rastreados. Resultados Se observó la predominancia de personas de sexo masculino, de raza blanca, de 60 años o más. La incidencia de eventos adversos acumulada fue del 4,4 % (n=13), todos fueron considerados como evitables y caracterizados de gravedad baja, en su mayoría. De acuerdo con la naturaleza, el 81,2 % de los eventos adversos estuvo relacionado con el cuidado en general. Por otro lado, con relación a los factores intrínsecos y extrínsecos, se observó un promedio de 1,17 y 1,5 por paciente, respectivamente. Se constató que 17 pacientes fueron identificados con evento adverso potencial, con 22 incidentes en el análisis. Conclusión Se identificó que el 100 % de los eventos adversos era evitable y la mayoría relacionados con el cuidado en general. Estos resultados refuerzan la necesidad de analizar y comprender la incidencia y la evitabilidad de los eventos adversos en unidades de pronta atención, lo que permite la planificación e implementación de estrategias que busquen la elaboración de acciones de control y respuesta a estos eventos, y así garantizar una atención segura y de calidad.

Abstract Objective To analyze the incidence and avoidability of adverse events in adult patients hospitalized in an emergency care unit in the countryside of Minas Gerais. Methods A cohort study, with a retrospective review of 296 medical records using screening (phase 1) and evaluation (phase 2) forms, completed by non-medical professionals and physicians, respectively. In the first phase, the demographic data, clinical condition of the patients, potential adverse events, and the verification of the quality of the information recorded in the medical records were evaluated. In the second phase, previously tracked adverse events were identified and characterized. Results There was a predominance of men, white, aged 60 or older. The cumulative incidence of adverse events was 4.4% (n=13), all of which were considered preventable, and mostly characterized as mild in severity. According to nature, 81.2% of adverse events were related to care in general. In relation to intrinsic and extrinsic factors, an average of 1.17 and 1.5 per patient, respectively, was found. It was found that a total of 17 patients were identified with potential adverse events, with 22 incidents in the analysis. Conclusion It was identified that 100% of adverse events were preventable and most related to general care. These findings reinforce the need to assess and understand the incidence and preventability of adverse events in emergency care units, which allows for the planning and implementation of strategies aimed at the development of control and response actions to these events, ensuring safe and quality care.

Posteromedial thalamic nucleus activity significantly contributes to perceptual discrimination.

Higher-order sensory thalamic nuclei are densely connected with multiple cortical and subcortical areas, yet the role of these nuclei remains elusive. The posteromedial thalamic nucleus (POm), the higher-order thalamic nucleus in the rodent somatosensory system, is an anatomical hub broadly connected with multiple sensory and motor brain areas yet weakly responds to passive sensory stimulation and whisker movements. To understand the role of POm in sensory perception, we developed a self-initiated, two-alternative forced-choice task in freely moving mice during active sensing. Using optogenetic and chemogenetic manipulation, we show that POm plays a significant role in sensory perception and the projection from the primary somatosensory cortex to POm is critical for the contribution of POm in sensory perception during active sensing.

Aplicação do Medical Office Survey on Patient Safety Culture: revisão integrativa / Aplicación del Medical Office Survey on Patient Safety Culture: revisión integradora / Application of the Medical Office Survey on Patient Safety Culture: integrative review

Resumo Objetivo Verificar como se expressa a cultura da Segurança do Paciente na visão de profissionais da Atenção Primária à Saúde, a partir da análise de produções científicas que aplicaram o instrumento Medical Office Survey on Patient Culture. Métodos Revisão integrativa, elaborada com base na metodologia scoping review, com busca em banco de dados online da Biblioteca Virtual em Saúde, da Web of Science, do PubMed®, do Cummulative Index to Nursing and Allied Health Literature e do Scopus, utilizando a palavra-chave "Medical Office Survey on Patient Safety Culture". Resultados Foram analisados 13 artigos quanto à aplicação do instrumento. Constatou-se avaliação geral positiva sobre Segurança do Paciente (32% a 83%). As dimensões "trabalho em equipe" e "pressão e ritmo de trabalho" foram vistas como as de melhor e pior escore, respectivamente. Conclusão A revisão integrativa da literatura possibilitou a análise crítica de estudos que demonstram a visão dos profissionais de saúde de diferentes países, ao avaliarem dimensões de cultura de Segurança do Paciente conforme o instrumento aplicado, indicando áreas consideradas positivas, bem como aquelas que demandam maior atenção e valorização. Tais evidências contribuem para o avanço na compreensão do multifacetado fenômeno investigado nos diversos ambientes da Atenção Primária.

Resumen Objetivo Verificar cómo se expresa la cultura de la Seguridad del Paciente bajo la mirada de profesionales de la Atención Primaria en Salud, a partir del análisis de producciones científicas que aplicaron el instrumento Medical Office Survey on Patient Culture. Métodos Revisión integradora, elaborada con base en la metodología scoping review y búsqueda en banco de datos en línea de la Biblioteca Virtual de Salud, de la Web of Science, de PubMed®, del Cummulative Index to Nursing and Allied Health Literature y de Scopus, utilizando la palabra clave "Medical Office Survey on Patient Safety Culture". Resultados Se analizaron 13 artículos con relación a la aplicación del instrumento. Se constató una evaluación general positiva sobre la Seguridad del Paciente (32 % a 83 %). Las dimensiones "trabajo en equipo" y "presión y ritmo de trabajo" fueron las que tuvieron mejor y peor puntuación, respectivamente. Conclusión La revisión integradora de la literatura posibilitó el análisis crítico de estudios que demuestran la visión de los profesionales de salud de distintos países, al evaluar las dimensiones de la cultura de la Seguridad del Paciente de acuerdo con el instrumento aplicado, indicando áreas consideradas positivas, así como las que demandan más atención y valorización. Esas evidencias contribuyen para el avance de la comprensión del multifacético fenómeno investigado en los diversos ambientes de la Atención Primaria.

Abstract Objective To assess how the Patient Safety culture is expressed in the view of Primary Health Care professionals, based on the analysis of scientific productions in which the Medical Office Survey on Patient Culture instrument was applied. Methods Integrative review based on the scoping review methodology with search in online databases of the Virtual Health Library, Web of Science, PubMed®, Cumulative Index to Nursing and Allied Health Literature and Scopus using the keyword "Medical Office Survey on Patient Safety Culture". Results Thirteen articles were analyzed in relation to the application of the instrument. There was a positive overall assessment of Patient Safety (32%-83%). The "teamwork" and "work pressure and pace" dimensions were seen as the best and worst scores, respectively. Conclusion The integrative literature review allowed the critical analysis of studies that showed the view of health professionals from different countries when evaluating dimensions of the Patient Safety culture according to the instrument applied, indicating areas considered as positive, and those demanding greater attention and appreciation. Such evidence contributes to advance the understanding of the multifaceted phenomenon investigated in different Primary Care settings.

Propuesta teórica para tratar los aspectos conductuales de la incontinencia urinaria.

Theoretical model for treating behavioural aspects of urinary incontinence SINOPSIS This reflective article presents a theoretical model of behavioural evaluation and intervention for women with urinary incontinence. Nola Pender's health promotion model (HPM) was used as a reference to develop this proposal. Behavioural measures were identified to build the model: constipation control, bladder training, urination position, reduction of irritating drinks, water intake, and pelvic floor muscle training. Next, specific actions were defined to investigate and intervene on certain behavioural aspects: previous related behaviour, personal factors, benefits, and obstacles regarding the proposed action, perceived self-efficacy, feelings related to the expected behaviour and the proposed action, interpersonal and situational influences, commitment to the action plan, competitive demands, and health promotion behaviour. The model proposed by the authors could help nurses to guide the conversation between them and the patient to modify certain behavioural aspects, to develop an achievable plan and, thus, enhance the achievement of the goals established for the treatment of urinary incontinence.

Sinopsis Este artículo de reflexión presenta un modelo teórico de evaluación e intervención conductual para mujeres con incontinencia urinaria (IU). El Modelo de Promoción de Salud de Nola Pender, enfermera estadounidense, fue usado como referencia para desarrollar la propuesta. Se identificaron medidas conductuales para construir el modelo: control del estreñimiento, entrenamiento vesical, posición para orinar, reducción de bebidas irritantes, ingesta de agua, y entrenamiento muscular del suelo pélvico. Luego, se definieron acciones determinadas para investigar e intervenir sobre ciertos aspectos conductuales: comportamiento previo relacionado, factores personales, beneficios y obstáculos percibidos acerca de la acción propuesta, autoeficacia percibida, sentimientos relacionados con el comportamiento esperado y la acción propuesta, influencias interpersonales y situacionales, compromiso con el plan de acción, exigencias competitivas, y comportamiento de la promoción de la salud. El modelo propuesto por los autores podría ayudar a enfermeros y enfermeras a orientar la conversación entre ellos y el paciente para modificar determinados aspectos conductuales, con el fin de elaborar un plan alcanzable y, así, potenciar el logro de las metas establecidas para el tratamiento de la IU.

Sharp force fatalities: Differentiating homicide from suicide through a retrospective review (2012-2019) of autopsy findings in Lisbon (Portugal).

Sharp force fatalities may have a homicidal, suicidal or accidental manner of death. To aid in such differentiation this study aimed to identify medico-legal elements which were predictors of a given manner of death as well as to describe the characteristics of these deaths. A retrospective review was performed on all homicides and suicides due to sharp force injury admitted at the South Branch of the National Institute of Legal Medicine and Forensic Sciences between January 2012 and December 2019. Deaths with a performed external examination or forensic autopsy and with available demographic, circumstantial or necroscopic information were included. Each case was reviewed to collect said information and inferential analysis was employed with both parametric and non-parametric tests as well as binary logistic regression to identify independent predictors, with significance defined at α = 0.05. A total of 57 homicides and 20 suicides were identified, with the obtained demographic and circumstantial profile of the homicide victim being that of a young foreign male whose body was found outside home, with no weapon nearby and without a known psychiatric background. Homicides presented more prominently stab wounds, with these being conspicuous on the thorax and neck. Conversely suicides notably presented cut wounds, being critically present in the neck and upper limbs. Oblique thoracic stab wounds conveyed a homicidal death. Other findings that suggested homicide included the presence of clothing damage, additional traumatic lesions and injured lungs or bone/cartilage. Toxicologically, alcohol presence was associated with homicides while psychiatric drugs suggested suicide. The logistic regression identified the presence of additional traumatic lesions (OR 14.8, p = 0.032) and the absence of lethal neck (OR 0.109, p = 0.043) and lethal upper limb (OR 0.022, p = 0.015) wounds as independent autopsy predictors of a homicidal death. However, no single feature is infallible in establishing manner of death. To achieve a cogent conclusion, all investigative elements must be considered while attending to the specifics of each case.

Sepse em pacientes com lesão renal aguda severa / Sepsis in patients with severe acute renal injury

Objetivo: analisar a prevalência de sepse e mortalidade dos pacientes com lesão renal aguda em tratamentorenal substitutivo hemodialítico. Método: estudo quantitativo, transversal, com 44 pacientes. Os dados foramcoletados com questionários e em prontuários, analisados por meio de medidas descritivas (média, mediana,desvio padrão e porcentagens) e apresentados em tabelas. Resultados: as variáveis sociodemográficasrevelaram que 63,6% são do sexo masculino, com idade média 63,3 anos. A sepse foi a principal causa de lesãorenal aguda em 65,9% dos pacientes e a mortalidade foi de 79,5%. O tempo de internação na UTI foi 22,45 ±13,5 dias e o tempo total de hospitalização foi 31,30 ± 21,4 dias. A hemodiálise intermitente foi a maisutilizada em 61,4% dos casos. Conclusão: apesar de campanhas internacionais, a sepse e a sua mortalidadepermanecem altas.

Study of InDel genetic markers with forensic and ancestry informative interest in PALOP's immigrant populations in Lisboa.

The migratory phenomenon in Portugal has become one of the main factors for the genetic variability. In the last few years, a new class of autosomal insertion/deletion markers-InDel-has attracted interest in forensic genetics. Since there is no data for InDel markers of Portuguese-speaking African countries (PALOP) immigrants living in Lisboa, our aim is the characterization of those groups of individuals by typing them with at least 30 InDel markers and to compare different groups of individuals/populations. We studied 454 bloodstain samples belonging to immigrant individuals from Angola, Guinea-Bissau, and Mozambique. DNA extraction was performed with the Chelex® 100 method. After extraction, all samples were typed with the Investigator® DIPplex method. Through the obtained results, allelic frequencies show that all markers are at Hardy-Weinberg equilibrium, and we can confirm that those populations show significant genetic distances between themselves, between them, and the host Lisboa population. Because of this, they introduce genetic variability in Lisboa population.

Ruptura esplênica secundária à infeção por Plasmodium falciparum / Splenic rupture secondary to Plasmodium falciparum infection

A ruptura esplênica é uma complicação possível da malária. É importante pela dificuldade diagnóstica, pois um elevado índice de suspeição é necessário para um diagnóstico atempado. Pode condicionar uma hemorragia intraperitoneal e deve ser considerada no diagnóstico diferencial de quadros de dor abdominal, hipotensão e diminuição do hematócrito. Os autores descrevem o caso de um homem de 59 anos, com ruptura esplênica secundária à malária por Plasmodium falciparum, tendo realizado esplenectomia urgente. Com a apresentação do caso, os autores pretendem chamar a atenção para a necessidade de incluir esta afecção no diagnóstico diferencial dos doentes com malária e hipotensão refractária.

Splenic rupture is a possible complication of malaria. Due to its difficult diagnosis, it is important, because a high level of suspicion is needed for a timely diagnosis. It results in intraperitoneal bleeding and should be considered in the differential diagnosis of abdominal pain, hypotension and low hematocrit. The authors report the case of a 59-year old man with splenic rupture secondary to malarial infection by Plasmodium Falciparum, who was treated with urgent splenectomy. This case is presented to remind the clinicians of include this entity in the differential diagnosis of patients with malaria and refractory hypotension.

Sensory feedback synchronizes motor and sensory neuronal networks in the neonatal rat spinal cord.

Early stages of sensorimotor system development in mammals are characterized by the occurrence of spontaneous movements. Whether and how these movements support correlated activity in developing sensorimotor spinal cord circuits remains unknown. Here we show highly correlated activity in sensory and motor zones in the spinal cord of neonatal rats in vivo. Both during twitches and complex movements, movement-generating bursts in motor zones are followed by bursts in sensory zones. Deafferentation does not affect activity in motor zones and movements, but profoundly suppresses activity bursts in sensory laminae and results in sensorimotor uncoupling, implying a primary role of sensory feedback in sensorimotor synchronization. This is further supported by largely dissociated activity in sensory and motor zones observed in the isolated spinal cord in vitro. Thus, sensory feedback resulting from spontaneous movements is instrumental for coordination of activity in developing sensorimotor spinal cord circuits.

Trichobezoar - A Rare Cause of Abdominal Mass and Gastric Outlet Obstruction.

The authors present the clinical case of a 14-year old girl with weight loss, anorexia, epigastric abdominal pain and postprandial vomiting with 5 months duration. There was a background of trichophagia for 2 years without evidence of alopecia or psychiatric history. The physical examination revealed an epigastric mass motionless, stony, with poorly defined limits, painful on palpation and about 7 cm diameter. Abdominal ultrasonography showed thickening of the gastric wall and antrum with gastric distension. The abdominal tomography scan and endoscopic examination revealed the presence of a bulky trichobezoar occupying almost the entire gastric lumen. It was decided to undergo gastrotomy and extraction of the bezoar. The postoperative period was uneventful.

Apresentamos o caso clínico de uma adolescente de 14 anos com emagrecimento, anorexia, epigastralgia e vómitos pós-prandiais com 5 meses de evolução. Apurou-se história de tricofagia com 2 anos de evolução mas sem áreas de alopécia ou antecedentes psiquiátricos. Apresentava massa epigástrica imóvel, pétrea, de limites mal definidos, dolorosa à palpação e com cerca de 7 cm de diâmetro. A ecografia abdominal mostrou espessamento da parede gástrica e do antro com distensão gástrica. Realizou tomografia abdominal e endoscopia digestiva alta que revelaram a presença de volumoso tricobezoar ocupando quase a totalidade do lúmen gástrico. Foi decidido realizar gastrotomia e extração do bezoar. O pós-operatório decorreu sem incidentes.

Endogenous IFN-ß signaling exerts anti-inflammatory actions in experimentally induced focal cerebral ischemia.

BACKGROUND: Interferon (IFN)-ß exerts anti-inflammatory effects, coupled to remarkable neurological improvements in multiple sclerosis, a neuroinflammatory condition of the central nervous system. Analogously, it has been hypothesized that IFN-ß, by limiting inflammation, decreases neuronal death and promotes functional recovery after stroke. However, the core actions of endogenous IFN-ß signaling in stroke are unclear. METHODS: To address this question, we used two clinically relevant models of focal cerebral ischemia, transient and permanent middle cerebral artery occlusion, and two genetically modified mouse lines, lacking either IFN-ß or its receptor, the IFN-α/ß receptor. Subsets of inflammatory and immune cells isolated from the brain, blood, and spleen were studied using flow cytometry. Sensorimotor deficits were assessed by a modified composite neuroscore, the rotating pole and grip strength tests, and cerebral infarct volumes were given by lack of neuronal nuclei immunoreactivity. RESULTS: Here, we report alterations in local and systemic inflammation in IFN-ß knockout (IFN-ßKO) mice over 8 days after induction of focal cerebral ischemia. Notably, IFN-ßKO mice showed a higher number of infiltrating leukocytes in the brain 2 days after stroke. Concomitantly, in the blood of IFN-ßKO mice, we found a higher percentage of total B cells but a similar percentage of mature and activated B cells, collectively indicating a higher proliferation rate. The additional differential regulation of circulating cytokines and splenic immune cell populations in wild-type and IFN-ßKO mice further supports an important immunoregulatory function of IFN-ß in stroke. Moreover, we observed a significant weight loss 2-3 days and a reduction in grip strength 2 days after stroke in the IFN-ßKO group, while endogenous IFN-ß signaling did not affect the infarct volume. CONCLUSIONS: We conclude that endogenous IFN-ß signaling attenuates local inflammation, regulates peripheral immune cells, and, thereby, may contribute positively to stroke outcome.

Isoflurane suppresses early cortical activity.

OBJECTIVE: Isoflurane and other volatile anesthetics are widely used in children to induce deep and reversible coma, but they may also exert neurotoxic actions. The effects of volatile anesthetics on the immature brain activity remain elusive, however. METHODS: The effects of isoflurane on spontaneous and sensory-evoked activity were explored using intracortical extracellular field potential and multiple unit recordings in the rat barrel cortex from birth to adulthood. RESULTS: During the first postnatal week, isoflurane suppressed cortical activity in a concentration-dependent manner. At surgical anesthesia levels (1.5-2%), isoflurane completely suppressed the electroencephalogram and silenced cortical neurons. Although sensory potentials evoked by the principal whisker deflection persisted, sensory-evoked early gamma and spindle-burst oscillations were completely suppressed by isoflurane. Isoflurane-induced burst-suppression pattern emerged during the second postnatal week and matured through the first postnatal month. Bursts in adolescent and adult rats were characterized by activation of entire cortical columns with a leading firing of infragranular neurons, and were triggered by principal and adjacent whiskers stimulation, and by auditory and visual stimuli, indicating an involvement of horizontal connections in their generation and horizontal spread. INTERPRETATION: The effects of isoflurane on cortical activity shift from total suppression of activity to burst-suppression pattern at the end of the first postnatal week. Developmental emergence of bursts likely involves a development of the intracortical short-and long-range connections. We hypothesize that complete suppression of cortical activity under isoflurane anesthesia during the first postnatal week may explain neuronal apoptosis stimulated by volatile anesthetics in the neonatal rats.

GABA(A) receptor dephosphorylation followed by internalization is coupled to neuronal death in in vitro ischemia.

Cerebral ischemia is characterized by an early disruption of GABAergic neurotransmission contributing to an imbalance of the excitatory/inhibitory equilibrium and neuronal death, but the molecular mechanisms involved are not fully understood. Here we report a downregulation of GABA(A) receptor (GABA(A)R) expression, affecting both mRNA and protein levels of GABA(A)R subunits, in hippocampal neurons subjected to oxygen-glucose deprivation (OGD), an in vitro model of ischemia. Similar alterations in the abundance of GABA(A)R subunits were observed in in vivo brain ischemia. OGD reduced the interaction of surface GABA(A)R with the scaffold protein gephyrin, followed by clathrin-dependent receptor internalization. Internalization of GABA(A)R was dependent on glutamate receptor activation and mediated by dephosphorylation of the ß3 subunit at serine 408/409. Expression of phospho-mimetic mutant GABA(A)R ß3 subunits prevented receptor internalization and protected hippocampal neurons from ischemic cell death. The results show a key role for ß3 GABA(A)R subunit dephosphorylation in the downregulation of GABAergic synaptic transmission in brain ischemia, contributing to neuronal death. GABA(A)R phosphorylation might be a therapeutic target to preserve synaptic inhibition in brain ischemia.

Effects of the sigma-1 receptor agonist 1-(3,4-dimethoxyphenethyl)-4-(3-phenylpropyl)-piperazine dihydro-chloride on inflammation after stroke.

Activation of the sigma-1 receptor (Sig-1R) improves functional recovery in models of experimental stroke and is known to modulate microglia function. The present study was conducted to investigate if Sig-1R activation after experimental stroke affects mediators of the inflammatory response in the ischemic hemisphere. Male Wistar rats were subjected to transient occlusion of the middle cerebral artery (MCAO) and injected with the specific Sig-1R agonist 1-(3,4-dimethoxyphenethyl)-4-(3-phenylpropyl)piperazine dihydrochloride (SA4503) or saline for 5 days starting on day 2 after MCAO. Treatment did not affect the increased levels of the pro-inflammatory cytokines interleukin 1 beta (IL-1ß), tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), interleukin 4 (IL-4), interleukin 5 (IL-5), and interleukin 13 (IL-13) in the infarct core and peri-infarct area after MCAO. In addition, treatment with SA4503 did not affect elevated levels of nitrite, TNF-α and IL-1ß observed in primary cultures of microglia exposed to combined Hypoxia/Aglycemia, while the unspecific sigma receptor ligand 1,3-di-o-tolylguanidine (DTG) significantly decreased the production of nitrite and levels of TNF-α. Analysis of the ischemic hemisphere also revealed increased levels of ionized calcium binding adaptor molecule 1 (Iba1) levels in the infarct core of SA4503 treated animals. However, no difference in Iba1 immunoreactivity was detected in the infarct core. Also, levels of the proliferation marker proliferating cell nuclear antigen (PCNA) and OX-42 were not increased in the infarct core in rats treated with SA4503. Together, our results suggest that sigma-1 receptor activation affects Iba1 expression in microglia/macrophages of the ischemic hemisphere after experimental stroke but does not affect post-stroke inflammatory mediators.

Excitotoxicity downregulates TrkB.FL signaling and upregulates the neuroprotective truncated TrkB receptors in cultured hippocampal and striatal neurons.

Brain-derived neurotrophic factor (BDNF) plays an important role in neuronal survival through activation of TrkB receptors. The trkB gene encodes a full-length receptor tyrosine kinase (TrkB.FL) and its truncated (T1/T2) isoforms. We investigated the changes in TrkB protein levels and signaling activity under excitotoxic conditions, which are characteristic of brain ischemia, traumatic brain injury, and neurodegenerative disorders. Excitotoxic stimulation of cultured rat hippocampal or striatal neurons downregulated TrkB.FL and upregulated a truncated form of the receptor (TrkB.T). Downregulation of TrkB.FL was mediated by calpains, whereas the increase in TrkB.T protein levels required transcription and translation activities. Downregulation of TrkB.FL receptors in hippocampal neurons correlated with a decrease in BDNF-induced activation of the Ras/ERK and PLCγ pathways. However, calpain inhibition, which prevents TrkB.FL degradation, did not preclude the decrease in signaling activity of these receptors. On the other hand, incubation with anisomycin, to prevent the upregulation of TrkB.T, protected to a large extent the TrkB.FL signaling activity, suggesting that truncated receptors may act as dominant-negatives. The upregulation of TrkB.T under excitotoxic conditions was correlated with an increase in BDNF-induced inhibition of RhoA, a mediator of excitotoxic neuronal death. BDNF fully protected hippocampal neurons transduced with TrkB.T when present during excitotoxic stimulation with glutamate, in contrast with the partial protection observed in cells overexpressing TrkB.FL or expressing GFP. These results indicate that BDNF protects hippocampal neurons by two distinct mechanisms: through the neurotrophic effects of TrkB.FL receptors and by activation of TrkB.T receptors coupled to inhibition of the excitotoxic signaling.

Cleavage of the vesicular glutamate transporters under excitotoxic conditions.

Glutamate is loaded into synaptic vesicles by vesicular glutamate transporters (VGLUTs), and alterations in the transporters expression directly regulate neurotransmitter release. We investigated changes in VGLUT1 and VGLUT2 protein levels after ischemic and excitotoxic insults. The results show that VGLUT2 is cleaved by calpains after excitotoxic stimulation of hippocampal neurons with glutamate, whereas VGLUT1 is downregulated to a lower extent. VGLUT2 was also cleaved by calpains after oxygen/glucose deprivation (OGD), and downregulated after middle cerebral artery occlusion (MCAO) and intrahippocampal injection of kainate. In contrast, VGLUT1 was not affected after OGD. Incubation of isolated synaptic vesicles with recombinant calpain also induced VGLUT2 cleavage, with a little effect observed for VGLUT1. N-terminal sequencing analysis showed that calpain cleaves VGLUT2 in the C-terminus, at Asn(534) and Lys(542). The truncated GFP-VGLUT2 forms were found to a great extent in non-synaptic regions along neurites, when compared to GFP-VGLUT2. These findings show that excitotoxic and ischemic insults downregulate VGLUT2, which is likely to affect glutamatergic transmission and cell death, especially in the neonatal period when the transporter is expressed at higher levels.