Novel image analysis tool for rapid screening of cell morphology in preclinical animal models of disease.

The field of cell biology has seen major advances in both cellular imaging modalities and the development of automated image analysis platforms that increase rigor, reproducibility, and throughput for large imaging data sets. However, there remains a need for tools that provide accurate morphometric analysis of single cells with complex, dynamic cytoarchitecture in a high-throughput and unbiased manner. We developed a fully automated image-analysis algorithm to rapidly detect and quantify changes in cellular morphology using microglia cells, an innate immune cell within the central nervous system, as representative of cells that exhibit dynamic and complex cytoarchitectural changes. We used two preclinical animal models that exhibit robust changes in microglia morphology: (1) a rat model of acute organophosphate intoxication, which was used to generate fluorescently labeled images for algorithm development; and (2) a rat model of traumatic brain injury, which was used to validate the algorithm using cells labeled using chromogenic detection methods. All ex vivo brain sections were immunolabeled for IBA-1 using fluorescence or diaminobenzidine (DAB) labeling, images were acquired using a high content imaging system and analyzed using a custom-built algorithm. The exploratory data set revealed eight statistically significant and quantitative morphometric parameters that distinguished between phenotypically distinct groups of microglia. Manual validation of single-cell morphology was strongly correlated with the automated analysis and was further supported by a comparison with traditional stereology methods. Existing image analysis pipelines rely on high-resolution images of individual cells, which limits sample size and is subject to selection bias. However, our fully automated method integrates quantification of morphology and fluorescent/chromogenic signals in images from multiple brain regions acquired using high-content imaging. In summary, our free, customizable image analysis tool provides a high-throughput, unbiased method for accurately detecting and quantifying morphological changes in cells with complex morphologies.

Percepción de la sexualidad en personas mayores de 60 años: Medellín, Colombia / The perception of sexuality after the age of 60 years: Medellin, colombia

La sexualidad es inherente y acompaña durante todo el curso de la vida a los seres humanos, trasciende el acto sexual para abordar aspectos de orden físico, emocional y cultural. Objetivo. Analizar los factores asociados a la percepción de la sexualidad según características demográficas, emocionales y psicológicas en personas mayores de 60 años. Materiales y métodos. Se realizó un estudio cuantitativo, observacional analítico, con fuente de información primaria a 665 personas de 60 años y más de la ciudad de Medellín, en el año 2018. Se determinaron las características demográficas, emocionales y psicológicas que permitieran conocer su percepción sobre sexualidad. Se calcularon medidas de asociación X2 y como medidas epide- miológicas, razones de prevalencia crudas y ajustadas. Resultados. Las personas que nunca se sintieron tristes consideraron importante la sexualidad (69,4 %), los hombres con una pareja estable, que no se sienten solos, con apoyo social también la consideraron como algo importante. Conclusiones. La importancia en la sexualidad de la persona mayor está determinada por estados anímicos, de estabilidad emocional y de integración social.

Sexuality is inherent and accompanies human beings throughout the course of life, transcends the sexual act, to address aspects of physical, emotional and cultural order. Objective. To analyze the factors associ- ated with the perception of sexuality according to demographic, emotional and psychological characteristics in people over 60 years of age. Materials and methods. A quantitative, analytical, observational study was conducted, with a primary source of information, to 665 people aged 60 and over, from the city of Medellin, in 2018. The demographic, emotional and psychological characteristics that allowed to know their percep- tion about sexuality. X2 association measures were calculated and as epidemiological measures, crude and adjusted prevalence ratios. Results. People who never felt sad considered sexuality important (69.4%), men with a stable partner, who do not feel alone, with social support also considered it important. Conclusions. The importance in the sexuality of the older person is determined by moods, emotional stability and social integration.

Diferencias en el sentimiento de soledad entre adultos institucionalizados y no institucionalizados / Differences in the Feeling of Loneliness Among Institutionalized and Non-institutionalized Elders / Diferencia no sentimento de solidão entre adultos institucionalizados e não institucionalizados

Resumen Objetivo: Comparar los niveles de soledad entre la población adulta institucionalizada y la no institucionalizada. Metodología: Investigación exploratoria de corte. Se midió el nivel de soledad reportada, usando una escala previamente validada (ESTE) en un grupo de adultos no institucionalizados, conformado por 869 mayores de 55 años, asistentes a los grupos de la tercera edad de las diferentes comunas de Medellín, captados mediante un muestreo aleatorio por conglomerados; asimismo, se aplicó en un grupo de adultos institucionalizados de 55 y más años de edad, residentes en centros gerontológicos públicos y privados, para un total de 379 adultos. Se analizaron las variables sociodemográficas: género, estado civil, escolaridad y edad, usando las pruebas U de Mann-Whitney y Kruskal-Wallis, con una significancia de 0,05. Resultados: Se observó mayor puntaje de soledad familiar, social y conyugal en los adultos institucionalizados. La crisis de adaptación fue similar en ambos grupos. El puntaje de soledad fue significativamente diferente en los adultos no institucionalizados en las variables de estado civil, escolaridad y edad, mientras que en los institucionalizados solo se observó diferencias con el estado civil. El puntaje total de soledad fue mayor en quienes no tenían pareja y estaban institucionalizados. Conclusiones: El sentimiento de soledad fue mayor en adultos institucionalizados. La variable estado civil estuvo asociada con la soledad en ambos grupos.

Abstract Objective: to compare levels of loneliness between institutionalized and non-institutionalized populations. Methodology: exploratory research to measure the level of loneliness using a previously validated scale (este) in a group of non-institutionalized elders made up of 869 people over the age of 55 who attended elderly groups at one of Medellin´s neighborhoods, selected by random cluster sampling. The scale was also applied to a group of 379 institutionalized elders over the age of 55 residing at public or private gerontological centers. The sociodemographic variables analyzed were: gender, marital status, education and age. U tests by Mann-Whitney and Kruskal-Wallis, with significance of 0,05 were used for this research. Results: a higher score in family, social and marital loneliness was observed in institutionalized elders. The adaptation crisis was equal in both groups. The loneliness score of non-institutionalized adults was significantly different in the variables of marital status, education and age. Meanwhile, for the institutionalized adults, differences were only observed in the marital status variable. The total loneliness score was higher in institutionalized elders without a sentimental partner. Conclusions: the feeling of loneliness was overall higher in institutionalized elders. The marital status variable was associated with loneliness in both groups.

Resumo Objetivo: comparar os níveis de solidão entre a população institucionalizada e a não institucionalizada. Metodologia: pesquisa exploratória de corte. Foi possível medir a solidão, com uma escala previamente validada (este), em um grupo de adultos não institucionalizados, conformado por 869 pessoas com mais de 55 anos, participantes dos grupos da terceira idade de várias comunas de Medellín, captados através de amostragem aleatória por conglomerados. Da mesma maneira, aplicou-se em um grupo de adultos institucionalizados de 55 ou mais anos de idade, residentes de centros gerontológicos públicos e particulares, para um total de 379 adultos. Analisaram-se variáveis sociodemográficas: género, estado civil, educação e idade, mediante provas U de Mann-Whitney e Kruskal-Wallis, com significância de 0,05. Resultados: observer am-se mais pontos de solidão familiar, social e conjugal nos idosos institucionalizados. A crise de adaptação foi similar em ambos os grupos. Os pontos de solidão foram significativamente diferentes nos adultos não institucionalizados nas variáveis de estado civil, educação e idade. Enquanto aos institucionalizados, só observou-se diferença no estado civil. O total de pontos de solidão foi mais alto nas pessoas sem companheiros sentimentais e estavam institucionalizadas. Conclusões: o sentimento de solidão foi mais alto em adultos institucionalizados. A variável de estado civil esteve associada com a solidão em ambos os grupos.

Stimulation of the medial septum improves performance in spatial learning following pilocarpine-induced status epilepticus.

Temporal lobe epilepsy often leads to hippocampal sclerosis and persistent cognitive deficits, including difficulty with learning and memory. Hippocampal theta oscillations are critical in optimizing hippocampal function and facilitating plasticity. We hypothesized that pilocarpine-induced status epilepticus would disrupt oscillations and behavioral performance and that electrical neuromodulation to entrain theta would improve cognition specifically in injured rats. Rats received a pilocarpine (n=30) or saline injection (n=27) and unilateral bi-polar electrodes were implanted into the medial septum and hippocampus the following day. Hippocampal and septal theta were recorded in a Plexiglas box over the first week following implantation. Control and pilocarpine-treated rats were split into stimulation (continuous 7.7Hz, 80µA, 1ms pulse width) and non-stimulation groups for behavioral analysis. Continuous stimulation was initiated one-minute prior to and throughout an object exploration task (post-injury day seven) and again for each of six trials on the Barnes maze (post-injury days 12-14). There was a significant reduction in hippocampal theta power (p<0.05) and percentage of time oscillating in theta (p<0.05). In addition there was a significant decrease in object exploration in rats post-pilocarpine (p<0.05) and an impairment in spatial learning. Specifically, pilocarpine-treated rats were more likely to use random search strategies (p<0.001) and had an increase in latency to find the hidden platform (p<0.05) on the Barnes maze. Stimulation of the medial septum at 7.7Hz in pilocarpine-treated rats resulted in performance similar to shams in both the object recognition and Barnes maze tasks. Stimulation of sham rats resulted in impaired object exploration (p<0.05) with no difference in Barnes maze latency or strategy. In conclusion, pilocarpine-induced seizures diminished hippocampal oscillations and impaired performance in both an object exploration and a spatial memory task in pilocarpine-treated rats. Theta stimulation at 7.7Hz improved behavioral outcome on the Barnes maze task; this improvement in function was not related to a general cognitive enhancement, as shams did not benefit from stimulation. Therefore, stimulation of the medial septum represents an exciting target to improve behavioral outcome in patients with epilepsy.

Septohippocampal Neuromodulation Improves Cognition after Traumatic Brain Injury.

Traumatic brain injury (TBI) often results in persistent attention and memory deficits that are associated with hippocampal dysfunction. Although deep brain stimulation (DBS) is used to treat neurological disorders related to motor dysfunction, the effectiveness of stimulation to treat cognition remains largely unknown. In this study, adult male Harlan Sprague-Dawley rats underwent a lateral fluid percussion or sham injury followed by implantation of bipolar electrodes in the medial septal nucleus (MSN) and ipsilateral hippocampus. In the first week after injury, there was a significant decrease in hippocampal theta oscillations that correlated with decreased object exploration and impaired performance in the Barnes maze spatial learning task. Continuous 7.7 Hz theta stimulation of the medial septum significantly increased hippocampal theta oscillations, restored normal object exploration, and improved spatial learning in injured animals. There were no benefits with 100 Hz gamma stimulation, and stimulation of sham animals at either frequency did not enhance performance. We conclude, therefore, that there was a theta frequency-specific benefit of DBS that restored cognitive function in brain-injured rats. These data suggest that septal theta stimulation may be an effective and novel neuromodulatory therapy for treatment of persistent cognitive deficits following TBI.

A clinical decision rule to predict adult patients with traumatic intracranial haemorrhage who do not require intensive care unit admission.

OBJECTIVE: To derive a clinical decision rule to identify adult emergency department (ED) patients with traumatic intracranial haemorrhage (tICH) who are at low risk for requiring critical care resources during hospitalization. METHODS: This is a retrospective cohort study of patients (≥ 18 years) with tICH presenting to the ED. The need for intensive care unit (ICU) admission was defined as the presence of a critical care intervention including: intubation, neurosurgical intervention, blood product transfusion, vasopressor or inotrope administration, invasive monitoring for haemodynamic instability, emergent treatment for arrhythmia, therapeutic angiography, and cardiopulmonary resuscitation. The decision rule was derived using binary recursive partitioning. RESULTS: A total of 432 patients were identified (median age 48 years) of which 174 patients (40%) had a critical care intervention. We performed binary recursive partitioning with Classification and Regression Trees (CART) software to develop the clinical decision rule. Patients with a normal mental status (Glasgow Coma Score=15), isolated head injury, and age<65 were considered low risk for a critical care intervention. The derived rule had a sensitivity of 98% (95% confidence interval [CI] 94-99), a specificity of 50% (95% CI 44-56), a positive predictive value of 57% (95% CI 51-62), and a negative predictive value of 97% (95% CI 93-99). The area under the curve for the decision rule was 0.74 (95% CI 0.70-0.77). CONCLUSIONS: This clinical decision rule identifies low risk adult ED patients with tICH who do not need ICU admission. Further validation and refinement of these findings would allow for more appropriate ICU resource utilisation.

Validación de la escala ESTE para medir la soledad de la población adulta / Validação da escala ESTE para medir a solidão da população adulta / ESTE scale validation to measure loneliness in adult population

Objetivo. Validar la escala de medición de la soledad ESTE en población adulta que asiste a los grupos de la tercera edad del municipio de Bello (Colombia). Metodología. Estudio descriptivo de corte transversal. Se evaluó su consistencia interna, reproducibilidad y comportamiento con variables sociodemográficas. La muestra estuvo conformada por 180 adultos de 55 o más años pertenecientes a 15 grupos de la tercera edad elegidos aleatoriamente de los 100 registrados en Bello a la fecha de realización del estudio. Resultados. El análisis factorial de los cuatro componentes principales de la escala explican el 55.9% de la varianza. Los factores que más varianza explican son: crisis de adaptación (17.2%), soledad conyugal (14.0%) y soledad social (13,9%). La soledad familiar fue el factor que menos explicó (10.9%). La confiabilidad de la escala Alfa de Cronbach de 0.9, siendo en cada uno de los factores superior a 0.7. La reproducibilidad test– retest medido por el coeficiente de correlación de Spearman fue 0.88. Conclusión. La validación de la escala ESTE indica que se puede utilizar como instrumento de medición de la soledad para la población adulta colombiana.

Objective. To validate the loneliness measurement scale ESTE in adult population who assist to third age groups in Bello (Colombia). Methodology. Cross sectional descriptive study. Internal consistency, reproducibility and behavior were assessed using demographic variables. The sample consisted of 180 adults aged 55 or over belonging to 15 senior groups selected randomly from the 100 registered in Bello at the time of the study execution. Results. The factorial analysis of the scales’ four main components explained 55.9% of the variance. Factors that explain greater variance are: adaptation crisis (17.2%), marriage loneliness (14.0%), and social loneliness (13.9%). Family loneliness was the factor that Invest Educ Enferm 2010;28(3) / 417 less explained (10.9%). Cronbach Alfa scale reliability of 0.9, being in each of the factors above 0.7. The test-retest reproducibility measured by the Sperman correlation coefficient was 0.88. Conclusion. ESTE scale validation indicates that it can be used as an adult loneliness measurement instrument in Colombia’s adult population.

Objetivo. Validar a escala de medição da solidão ESTE em população adulta que assiste aos grupos da terceira idade do município de Bello (Colômbia). Metodologia. Estudo descritivo de corte transversal. Avaliou-se sua consistência interna, reprodutibilidade e comportamento com variáveis sócio-demográficas. A mostra esteve conformadas por 180 adultos de 55 ou mais anos pertencentes a 15 grupos da terceira idade eleitos aleatoriamente dos 100 registrados em Belo à data de realização do estudo. Resultados. A análise fatorial dos quatro componentes principais da escala explica o 55.9% da variância. Os fatores que mais variancia explicam são: crise de adaptação (17.2%), solidão conjugal (14.0%) e solidão social (13,9%). A solidão familiar foi o fator que menos explicou (10.9%). A confiança da escala Alfa de Cronbach de 0.9, sendo em cada um dos fatores superior a 0.7. A reprodutibilidade teste–reteste medido pelo coeficiente de correlação de Spearman foi 0.88. Conclusão. A validação da escala ESTE indica que se pode utilizar como instrumento de medição da solidão para a população adulta colombiana.

Regulation of poliovirus 3C protease by the 2C polypeptide.

Poliovirus-encoded nonstructural polypeptide 2C is a multifunctional protein that plays an important role in viral RNA replication. 2C interacts with both intracellular membranes and virus-specific RNAs and has ATPase and GTPase activities. Extensive computer analysis of the 2C sequence revealed that in addition to the known ATPase-, GTPase-, membrane-, and RNA-binding domains it also contains several "serpin" (serine protease inhibitor) motifs. We provide experimental evidence suggesting that 2C is indeed capable of regulating virus-encoded proteases. The purified 2C protein inhibits 3C(pro)-catalyzed cleavage of cellular transcription factors at Q-G sites in vitro. It also inhibits cleavage of a viral precursor by the other viral protease, 2A(pro). However, at least three cellular proteases appear not to be inhibited by 2C in vitro. The 2C-associated protease inhibitory activity can be depleted by anti-2C antibody. A physical interaction between 2C and His-tagged 3C(pro) can be demonstrated in vitro by coimmunoprecipitation of 2C with anti-His antibody. Deletion analysis suggests that the 2C central and C-terminal domains that include several serpin motifs are important for 3C(pro)-inhibitory activity. To examine the 2C protease inhibitory activity in vivo, stable HeLa cell lines were made that express 2C in an inducible fashion. Infection of 2C-expressing cells with poliovirus led to incomplete (or inefficient) processing of viral precursor polypeptides compared to control cell lines containing the vector alone. These results suggest that 2C can negatively regulate the viral protease 3C(pro). The possible role of the 2C protease inhibitory activity in viral RNA replication is discussed.

Immune dysfunction and the pathogenesis of AIDS-associated non-Hodgkin's lymphoma

Much has been learned about how HIV-induced immune dysfunction contributes to B cell hyperactivation, and potentially, to the pathogenesis of AIDS-lymphoma. However, further studies are needed to fully understand how HIV infection and immune dysfunction promote B cell hyperactivation and the development/growth of AIDS-lymphoma. In particular, studies are needed to define the role of HHV8 vIL6, IL6 receptor-expression, and lymphocyte surface stimulatory molecules, in promoting B cell hyperactivation or lymphoma cell growth.