Treatment of esophageal perforation: A review of our experience at a tertiary referral hospital spanning the past 19 years.

INTRODUCTION AND AIMS: Esophageal perforation is an uncommon event that is a medical/surgical emergency, with a 15-30% mortality rate. The aim of the present study was to communicate our experience in the management of esophageal perforation, evaluating the different strategies utilized, in an effort to establish measures to guide decision-making in selecting treatment. MATERIALS AND METHODS: A retrospective descriptive study was conducted on patients diagnosed with esophageal perforation at our hospital center, within the time frame of 2000 and 2019. RESULTS: Over the past 19 years, 15 patients were diagnosed with esophageal perforation. Surgical treatment was carried out in 80% of the cases. Primary closure, reinforced with plasty, was performed in 67% of the patients, of whom 62.5% had early diagnosis and a 100% survival rate. Diagnosis was late in 37.5% of the cases, with a 33.3% survival rate. Esophagectomy and gastric pull-up were performed on 25% of the patients, 66.6% of whom had early diagnosis and a 100% survival rate. In the 33.3% that had late diagnosis, the mortality rate was 100%. Esophagectomy, with cervical esophagostomy and feeding jejunostomy, was performed on one of the patients (8.3%) that had early diagnosis and a 100% survival rate. CONCLUSIONS: The main survival predictor in esophageal perforation is the interval of time between the injury and its diagnosis, and in turn, the resulting treatment. Each patient with esophageal perforation should have individualized treatment to adequately manage the condition.

Genetic and cytological analyses reveal the recombination landscape of a partially differentiated plant sex chromosome in kiwifruit.

BACKGROUND: Angiosperm sex chromosomes, where present, are generally recently evolved. The key step in initiating the development of sex chromosomes from autosomes is the establishment of a sex-determining locus within a region of non-recombination. To better understand early sex chromosome evolution, it is important to determine the process by which recombination is suppressed around the sex determining genes. We have used the dioecious angiosperm kiwifruit Actinidia chinensis var. chinensis, which has an active-Y sex chromosome system, to study recombination rates around the sex locus, to better understand key events in the development of sex chromosomes. RESULTS: We have confirmed the sex-determining region (SDR) in A. chinensis var. chinensis, using a combination of high density genetic mapping and fluorescent in situ hybridisation (FISH) of Bacterial Artificial Chromosomes (BACs) linked to the sex markers onto pachytene chromosomes. The SDR is a subtelomeric non-recombining region adjacent to the nucleolar organiser region (NOR). A region of restricted recombination of around 6 Mbp in size in both male and female maps spans the SDR and covers around a third of chromosome 25. CONCLUSIONS: As recombination is suppressed over a similar region between X chromosomes and between and X and Y chromosomes, we propose that recombination is suppressed in this region because of the proximity of the NOR and the centromere, with both the NOR and centromere suppressing recombination, and this predates suppressed recombination due to differences between X and Y chromosomes. Such regions of suppressed recombination in the genome provide an opportunity for the evolution of sex chromosomes, if a sex-determining locus develops there or translocates into this region.

Cobertura Universal de Servicios Diagnósticos a través de la Telemedicina / Universal Coverage of Diagnostic Services through Telemedicine

Con la Telemedicina pueden desarrollarse sistemas de telediagnóstico ventajosos para mejorar la atención de la salud de poblaciones remotas que no tienen acceso a especialistas. Este estudio realizado por la Unidad de Telemedicina del Ministerio de Salud Pública y Bienestar Social (MSPBS) en colaboración con el Dpto. de Ingeniería Biomédica e Imágenes del Instituto de Investigaciones en Ciencias (IICS-UNA) y la Universidad del País Vasco (UPV/EHU) sirvió para evaluar la utilidad de un sistema de telediagnóstico para la cobertura universal en la salud pública. Para el efecto se analizaron los resultados obtenidos por el sistema de telediagnóstico implementado en 56 hospitales del MSPBS. En dicho sentido se analizaron 293.142 diagnósticos remotos realizados entre enero del 2014 y septiembre de 2017. Del total, el 37,29 % (109.311) correspondieron a estudios de tomografía, 61,44 % (180.108) a electrocardiografía (ECG), 1,26 % (3.704) a electroencefalografía (EEG) y 0,01 % (19) a ecografía. No se observaron diferencias significativas entre el diagnóstico remoto y el diagnóstico "cara a cara". Con el diagnostico remoto se logró una reducción del coste que supone un beneficio importante para cada ciudadano del interior del país. Los resultados obtenidos evidencian que la telemedicina puede contribuir para mejorar significativamente la cobertura universal de los servicios diagnósticos y programas de salud, maximizando el tiempo del profesional y su productividad, aumentando el acceso y la equidad, y disminuyendo los costos. Sin embargo antes de realizar su implementación sistemática se deberá realizar una contextualización con el perfil epidemiológico regional. Palabras claves: Telemedicina; Cobertura Universal; Telediagnóstico; TICs en Salud; Innovación Tecnológica.

Through the telemedicine, advantageous telediagnostic systems can be developed to improve the health care of remote populations that don`t have access to specialists. This study was carried out by the Telemedicine Unit of the Ministry of Public Health and Social Welfare (MSPBS) in collaboration with the Department of Biomedical Engineering and Imaging of the Health Science Research Institute (IICS-UNA) and the University of the Basque Country (UPV / EHU) to evaluate the utility of a telediagnostic system for universal coverage in public health. For this purpose, the results obtained by the telediagnosis system implemented in 56 MSPBS hospitals were analyzed. In that sense, 293,142 remote diagnoses were performed between January 2014 and September 2017. Of the total, 37.29% (109,311) corresponded to tomography studies, 61.44% (180.108) to electrocardiography (ECG), 1.26% (3,704) to electroencephalography (EEG) and 0.01% (19) to ultrasound. There were no significant differences between the remote and the "face to face" diagnosis. With the remote diagnosis a reduction of the cost was obtained, that supposes an important benefit for each citizen of the interior of the country. The results show that the telemedicine can contribute to improve significantly the universal coverage of diagnostic services and health programs, maximizing professional time and productivity, increasing access and equity, and reducing costs. However, before carrying out its systematic implementation, a contextualization with the regional epidemiological profile must be performed. Key words: Telemedicine; Universal Coverage; Telediagnosis; ICT in Health; Technological Innovation.

Characterization of Gene Expression in the Rat Brainstem After Neonatal Hypoxic-Ischemic Injury and Antioxidant Treatment.

The perinatal brainstem is known to be very vulnerable to hypoxic-ischemic events which can lead to deafness, swallowing dysfunction, and defective respiratory control. The aim of the present work was to evaluate the potential neuroprotective effects of nicotine, melatonin, resveratrol, and docosahexaenoic acid on the expression of a panel of genes in the brainstem following hypoxic-ischemic damage. Quantitative PCR was used to examine gene expression 3 and 12 h after the damage, and immunohistochemistry was employed to evaluate neurons, astrocytes, and synaptic vesicles 24 h post insult. We found that the expression of some immediate-early genes, as well as that of inflammatory genes TNF-α, COX2, and caspase 3, was upregulated in response to the insult. Twenty-four hours after the damage, the percentage of NeuN and synaptophysin immunolabeled cells was found to be reduced while GFAP expression was upregulated. No differences were observed in ROS gene expression following treatments.

Docosahexaenoic Acid Reduces Cerebral Damage and Ameliorates Long-Term Cognitive Impairments Caused by Neonatal Hypoxia-Ischemia in Rats.

As the interest in the neuroprotective possibilities of docosahexaenoic acid (DHA) for brain injury has grown in the recent years, we aimed to investigate the long-term effects of this fatty acid in an experimental model of perinatal hypoxia-ischemia in rats. To this end, motor activity, aspects of learning, and memory function and anxiety, as well as corticofugal connections visualized by using tracer injections, were evaluated at adulthood. We found that in the hours immediately following the insult, DHA maintained mitochondrial inner membrane integrity and transmembrane potential, as well as the integrity of synaptic processes. Seven days later, morphological damage at the level of the middle hippocampus was reduced, since neurons and myelin were preserved and the astroglial reactive response and microglial activation were seen to be diminished. At adulthood, the behavioral tests revealed that treated animals presented better long-term working memory and less anxiety than non-treated hypoxic-ischemic animals, while no difference was found in the spontaneous locomotor activity. Interestingly, hypoxic-ischemic injury caused alterations in the anterograde corticofugal neuronal connections which were not so evident in rats treated with DHA. Thus, our results indicate that DHA treatment can lead to long-lasting neuroprotective effects in this experimental model of neonatal hypoxia-ischemic brain injury, not only by mitigating axonal changes but also by enhancing cognitive performance at adulthood.

Measuring affiliation in group therapy for substance use disorders in the Women's Recovery Group study: Does it matter whether the group is all-women or mixed-gender?

BACKGROUND AND OBJECTIVES: A Stage II, two-site randomized clinical trial compared the manualized, single-gender Women's Recovery Group (WRG) to mixed-gender group therapy (Group Drug Counseling; GDC) and demonstrated efficacy. Enhanced affiliation and support in the WRG is a hypothesized mechanism of efficacy. This study sought to extend results of the previous small Stage I trial that showed the rate of supportive affiliative statements occurred more frequently in WRG than GDC. METHODS: Participants (N = 158; 100 women, 58 men) were 18 years or older, substance dependent, and had used substances within the past 60 days. Women were randomized to WRG (n = 52) or GDC (n = 48). Group therapy videos were coded by two independent raters; Rater 1 coded 20% of videos (n = 74); Rater 2 coded 25% of videos coded by Rater 1 (n = 19). RESULTS: The number of affiliative statements made in WRG was 66% higher than in GDC. Three of eight affiliative statement categories occurred more frequently in WRG than GDC: supportive, shared experience, and strategy statements. DISCUSSION AND CONCLUSIONS: This larger Stage II trial provided a greater number of group therapy tapes available for analysis. Results extended our previous findings, demonstrating both greater frequency of all affiliative statements, as well as specific categories of statements, made in single-gender WRG than mixed-gender GDC. SCIENTIFIC SIGNIFICANCE: Greater frequency of affiliative statements among group members may be one mechanism of enhanced support and efficacy in women-only WRG compared with standard mixed-gender group therapy for substance use disorders. (Am J Addict 2016;25:573-580).

Costo-Beneficio de un Sistema de Telediagnóstico para Hospitales Regionales y Distritales del Paraguay / Cost-Effectiveness of a Remote Telediagnostic System for Regional and District Hospitals in Paraguay

En el contexto de una evaluación económica y tecnológica de las alternativas metodológicas que facilite un sistema de cobertura universal y el uso eficiente de los recursos disponibles en la salud pública, existen argumentos de costo-beneficio para que un sistema de telediagnóstico sea considerado ventajoso en los países en vías de desarrollo como una herramienta para mejorar la atención de la salud de poblaciones remotas que no tienen acceso a los especialistas. Este estudio observacional y descriptivo realizado por la Unidad de Telemedicina del Ministerio de Salud Pública y Bienestar Social (MSPBS) en colaboración con el Dpto. de Ingeniería Biomédica e Imágenes del Instituto de Investigaciones en Ciencias de la Salud de la Universidad Nacional de Asunción (IICS-UNA) y la Universidad del País Vasco (UPV/EHU) sirvió como un proyecto piloto para evaluar el costo-beneficio de un sistema de telediagnóstico en la salud pública. Para el efecto fueron analizados los resultados obtenidos en un proyecto piloto de telediagnóstico implementado en 25 hospitales regionales y distritales del MSPBS. En dicho sentido, en el marco del proyecto piloto fueron realizados 34.096 diagnósticos remotos entre enero del 2014 y mayo de 2015 a través del sistema. Del total de telediagnósticos realizados, el 38,0 % (12.966) correspondieron a estudios de tomografía, 61,1 % (21.111) a electrocardiografía (ECG) y 0,1 % (19) a ecografía. Se observó una diferencia importante en el coste de diagnóstico remoto en relación al diagnóstico "cara a cara", en el análisis se incorporaron los costos de implantación y mantenimiento de la TIC para el diagnóstico remoto y los costos de transporte, alimentación y oportunidad para el diagnóstico "cara a cara". La reducción del coste a través del diagnóstico remoto fue de 4,5 veces para ECG, 26,4 veces para tomografía y de 8,3 veces para ecografía, lo que supone un beneficio importante para cada ciudadano del interior del país toda vez que el coste promedio de diagnóstico remoto para cada hospital sea igual o inferior al coste total del diagnóstico cara a cara. Los resultados obtenidos en este estudio contribuirán en forma significativa al fortalecimiento de la cobertura universal de servicios diagnósticos, capacidad de innovación y aseguramiento de la sostenibilidad económica del sistema de telediagnóstico público. Además ayudará a mejorar sustancialmente la capacidad resolutiva local de los hospitales regionales y distritales en el interior del país. Sin embargo, antes de recomendar su utilización masiva se deberá analizar los costos para su implementación y la sostenibilidad técnico-económica del sistema acorde a los recursos disponibles. Palabras claves: Telediagnóstico, Telesalud, Telemedicina, TICs en salud, Telemática en salud.

In the context of an assessment of alternative economic and technological methods that would facilitate a universal health coverage system and the effcient use of available resources in public health, there is debate about the cost-effectiveness and advantages of a remote telediagnostic system in developing countries as a tool to improve healthcare for remote populations without access to specialists. This observational and descriptive study by the Telemedicine Unit of the Ministry of Public Health and Social Welfare (MSPBS) in collaboration with the Dept. Of Biomedical Engineering and Imaging Research of the Institute in Health Sciences of the National University of Asuncion (IICS -A) and the University of the Basque Country (UPV / EHU) served as a pilot project to assess the cost-effectiveness of a system of remote telediagnostics in public health. For this purpose, we analyzed the results of a telediagnosis pilot project implemented in 25 regional and district MSPBS hospitals MSPBS. During the pilot project timeframe, 34,096 remote telediagnoses were made between January 2014 and May 2015 throughout the system. Of the total remote telediagnoses performed, 38.0% (12,966) were of tomography studies, 61.1% (21,111) were of electrocardiography (ECG) and 0.1% (19) were ultrasound interpretations. A signifcant difference was observed in the cost of remote telediagnosis compared to"face to face" diagnosis; the analysis included implementation and maintenance costs of information technology for remote telediagnoses vs. transportation costs, meals and access options for "face to face" diagnoses. The cost reduction via remote diagnosis was 4.5 times for ECG, 26.4 times for CT scans and 8.3 times for ultrasound, which would be a major beneft for patients living in remote areas of the country, since the average cost for remote telediagnosis at each hospital is equal to or less than the total cost of "fact to face" diagnosis. The results obtained in this study are a signifcant contribution to providing universal coverage for diagnostic services, improve the capacity to innovate in the public health system and reassure policymakers regarding the economic sustainability of a public health remote telediagnosis system. It will also help to substantially improve the resolution ability of regional and district hospitals in the countryside. However, before recommending its extensive use, the implementation costs, as well as the technical and economic sustainability considering local resource availability, should be analyzed. Keywords: Remote diagnostics, telehealth, telemedicine, health information technology, health Telematics.

Initial response as a predictor of 12-week buprenorphine-naloxone treatment response in a prescription opioid-dependent population.

OBJECTIVE: Initial medication response has been shown to predict treatment outcome across a variety of substance use disorders, but no studies have examined the predictive power of initial response to buprenorphine-naloxone in the treatment of prescription opioid dependence. We therefore conducted a secondary analysis of data from the Prescription Opioid Addiction Treatment Study to determine whether initial response to buprenorphine-naloxone predicted 12-week treatment outcome in a prescription opioid-dependent population. METHOD: Using data from a multisite, randomized controlled trial of buprenorphine-naloxone plus counseling for DSM-IV prescription opioid dependence (June 2006-July 2009), we conducted a secondary analysis to investigate the relationship between initial medication response and 12-week treatment outcome to establish how soon the efficacy of buprenorphine-naloxone could be predicted (N = 360). Outcomes were determined from the Substance Use Report, a self-report measure of substance use, and confirmatory urinalysis. Predictive values were calculated to determine the importance of abstinence versus use at various time points within the first month of treatment (week 1, weeks 1-2, 1-3, or 1-4) in predicting successful versus unsuccessful treatment outcome (based on abstinence or near-abstinence from opioids) in the last 4 weeks of buprenorphine-naloxone treatment (weeks 9-12). RESULTS: Outcome was best predicted by medication response after 2 weeks of treatment. Two weeks of initial abstinence was moderately predictive of treatment success (positive predictive value = 71%), while opioid use in both of the first 2 weeks was strongly predictive of unsuccessful treatment outcome (negative predictive value [NPV] = 84%), especially when successful outcome was defined as total abstinence from opioids in weeks 9-12 (NPV = 94%). CONCLUSIONS: Evaluating prescription opioid-dependent patients after 2 weeks of buprenorphine-naloxone treatment may help determine the likelihood of successful outcome at completion of the current treatment regimen. TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT00316277.

Denial of urinalysis-confirmed opioid use in prescription opioid dependence.

Although research has generally supported the validity of substance use self-reports, some patients deny urine-verified substance use. We examined the prevalence and patterns of denying urinalysis-confirmed opioid use in a sample of prescription opioid dependent patients. We also identified characteristics associated with denial in this population of increasing public health concern. Opioid use self-reports were compared with weekly urinalysis results in a 12-week multi-site treatment study for prescription opioid dependence. Among those who used opioids during the trial (n=246/360), 44.3% (n=109) denied urinalysis-confirmed opioid use, although usually only once (78%). Overall, 22.9% of opioid-positive urine tests (149/650) were denied on self-report. Multivariable analysis found that initially using opioids to relieve pain was associated with denying opioid use. These findings support the use of both self-reports and urine testing in treating prescription opioid dependence.

Perspectivas de un Sistema de Telemedicina en la Salud Pública del Paraguay. Estudio Piloto / Perspectives on a Telemedicine System in Public Health in Paraguay. A Pilot Study

En el contexto de una cobertura universal y el uso eficiente de los recursos disponibles en la salud pública que deben estar orientados hacia una mayor equidad en la prestación de servicios, mayor preocupación por la efectividad y utilidad de las tecnologías para la salud, existe una perspectiva favorable para que la telemedicina sea considerada en los países industrializados y en vías de desarrollo como una herramienta para mejorar la atención de la salud de poblaciones remotas que no tienen acceso a los especialistas. Este estudio observacional y descriptivo realizado por la Unidad de Telemedicina del Ministerio de Salud Pública y Bienestar Social (MSPBS) en colaboración con el Dpto. de Ingeniería Biomédica e Imágenes del Instituto de Investigaciones en Ciencias de la Salud de la Universidad Nacional de Asunción (IICSUNA) y la Universidad del País Vasco (UPV/EHU) sirvió como un proyecto piloto para evaluar las potencialidades de un sistema de telemedicina en la salud pública. Para el efecto fueron analizados los resultados preliminares de un proyecto piloto de telediagnóstico implementado en algunos hospitales regionales y distritales del MSPBS. En dicho sentido, en el marco del proyecto piloto fueron realizados 15968 diagnósticos remotos entre enero y noviembre de 2014 a través del sistema. Del total de telediagnósticos realizados, el 43,9 % (7008) correspondieron a estudios de tomografía, 56,0 % (8941) a electrocardiografía (ECG) y 0,1 % (19) a ecografía. Las dificultades observadas se relacionaron: al recurso humano (capacitación de asistente técnico, reticencia a aceptar totalmente el telediagnóstico como una nueva herramienta) y el tecnológico (identificación de las señales disponibles, baja velocidad de la red interna y los modelos de aplicación referidos a standalone o web). Los resultados obtenidos a través de este estudio piloto son de vital importancia para la formulación de metodologías prácticas y viables para la implantación de un sistema de telemedicina que ayudará a mejorar sustancialmente la capacidad resolutiva local de los centros asistenciales a las poblaciones remotas y dispersas e intercambiar con mayor efectividad informaciones clínicas, administrativas y de capacitación del personal. Este estudio piloto basado en las tecnologías disponibles, las experiencias previas propias y foráneas muestra las potencialidades de un sistema de telemedicina en la salud pública. Pero, antes de recomendar su utilización masiva se deberá realizar un estudio exhaustivo y pormenorizado de los sistemas de salud, los costos para su implementación y la sustentabilidad del sistema acorde a las metodologías vigentes. Palabras claves: Telemedicina, Telesalud, Teleeducación, Telecuidado, Telemática en salud.

In the context of universal coverage and the efficient use of available resources in public health which should be directed towards greater equity in the provision of services, greater concern for the effectiveness and usefulness of health technologies, there is a favorable opportunity to develop telemedicine in both developing and industrialized countries as a tool to improve health care in remote locations without access to specialists. This observational and descriptive study, performed by the Telemedicine Unit of the Ministry of Public Health and Social Welfare (MOHSW) in collaboration with the Dept. Of Biomedical Engineering & Imaging Research Institute in Health Sciences of the National University of Asunción (IICS-UNA) and the University of the Basque Country (UPV / EHU) served as a pilot project to evaluate the potential of a telemedicine system in public health. For these purposes, we analyzed preliminary results of a pilot project using telemedicine for diagnosis implemented in some remote regional and district hospitals MSPBS. During the pilot project time period, 15968 remote diagnostics were conducted between January and November 2014 using the system. Of all remote diagnostics modalities performed, 43.9% (7008) corresponded to tomography studies, 56.0% (8941) to electrocardiography (ECG) and 0.1% (19) to ultrasound studies. The observed difficulties were: human resources (training of technical assistants, reluctance to fully accept remote diagnostics as a new tool) and technological (difficult identification of signals available, low-speed internet network and computer applications that were "standalone" or web-based). The results obtained from this pilot study are vital for the development of practical and workable methodologies for the implementation of a telemedicine system that will help to substantially improve local response capacity of health facilities to remote and dispersed populations and to more effectively exchange clinical information, and train administrative and staff personnel. This pilot study, which was based on available technology and previous experiences in other countries, shows the potential of a telemedicine system in public health. But, before recommending its widespread use there will need to be a thorough and detailed study of health systems, costs for implementation and sustainability of the system using current methodologies. Keywords: Telemedicine, Telehealth, Tele-education, telecare, Telematics health

Changes in sleep disruption in the treatment of co-occurring posttraumatic stress disorder and substance use disorders.

Sleep disruption appears not only to reflect a symptom of posttraumatic stress disorder (PTSD), but also a unique vulnerability for its development and maintenance. Studies examining the impact of psychosocial treatments for PTSD on sleep symptoms are few and no studies to date of which we are aware have examined this question in samples with co-occurring substance use disorders. The current study is a secondary analysis of a large clinical trial comparing 2 psychological treatments for co-occurring PTSD and substance use disorders. Women (N = 353) completed measures of PTSD at baseline, end of treatment, and 3-, 6-, and 12-month follow-ups. Results indicated that the prevalence of insomnia, but not nightmares, decreased during treatment, and that 63.8% of participants reported at least 1 clinical-level sleep symptom at the end of treatment. Improvement in sleep symptoms during treatment was associated with better overall PTSD outcomes over time, χ(2) (1) = 33.81, p < .001. These results extend the existing literature to suggest that residual sleep disruption following PTSD treatment is common in women with co-occurring PTSD and substance use disorders. Research on the benefits of adding sleep-specific intervention for those with residual sleep disruption in this population may be a promising future direction.

Effect of belimumab on vaccine antigen antibodies to influenza, pneumococcal, and tetanus vaccines in patients with systemic lupus erythematosus in the BLISS-76 trial.

OBJECTIVE: In patients with systemic lupus erythematosus (SLE), evidence suggests that most vaccines (except live-virus vaccines) are safe, although antibody response may be reduced. This substudy from the phase III, randomized, double-blind, placebo-controlled BLISS-76 trial evaluated the effects of belimumab on preexisting antibody levels against pneumococcal, tetanus, and influenza antigens in patients with SLE. METHODS: In BLISS-76, patients with autoantibody-positive, active SLE were treated with placebo or belimumab 1 or 10 mg/kg every 2 weeks for 28 days and every 28 days thereafter, plus standard SLE therapy, for 76 weeks. This analysis included a subset of patients who had received pneumococcal or tetanus vaccine within 5 years or influenza vaccine within 1 year of study participation. Antibodies to vaccine antigens were tested at baseline and Week 52, and percentage changes in antibody levels from baseline and proportions of patients maintaining levels at Week 52 were assessed. Antibody titers were also assessed in a small number of patients vaccinated during the study. RESULTS: Consistent with preservation of the memory B cell compartment with belimumab treatment, the proportions of patients maintaining antibody responses to pneumococcal, tetanus, and influenza antigens were not reduced. In a small group receiving influenza vaccine on study, antibody responses were frequently lower with belimumab, although titer levels were > 1:10 in all patients treated with 10 mg/kg and in the majority treated with 1 mg/kg. CONCLUSION: Treatment with belimumab did not affect the ability of patients with SLE to maintain antibody titers to previous pneumococcal, tetanus, and influenza immunizations. [ClinicalTrials.gov registration number NCT 00410384].

Histological study of the protective effect of melatonin on neural cells after neonatal hypoxia-ischemia.

To minimize as much as possible the neurological consequences from hypoxic-ischemic (HI) brain injury, neuroprotective strategies are urgently required. In this sense, there is growing interest in the neuroprotective potential of melatonin after perinatal asphyxia, due to its high efficacy, low toxicity and ready cross through the blood-brain barrier. Twenty six Wistar rats at postnatal day 7 were randomly assigned to: two hypoxic-ischemic groups: pups with the left common carotid artery ligated and then submitted to hypoxia (HI group) and animals that received a dose of 15 mg/kg melatonin just after the hypoxic-ischemic event and repeated twice with an interval of 24 hours (HI+MEL group). Pups without ischemia or hypoxia were used as controls (Sham group). Seven days after surgery, brains were collected and coronal sections Nissl-stained, TUNEL-labeled, or MBP- and GFAP-immunolabeled prior to determining brain infarct area, quantify surviving neurons and evaluate oligodendroglial injury and reactive astrogliosis. The number of surviving neurons showing a well preserved architecture in HI+MEL group was similar to that observed in the Sham group. Moreover, TUNEL-positive cells only appeared in the HI group. The ratio of left-to-right hemispheric MBP immunostaining showed a significant decrease in the HI group in comparison with Sham pups, which was restored after melatonin administration. Melatonin also reduced reactive gliosis. Thus, our results suggest that treatment with melatonin after neonatal hypoxia-ischemia led to a neuroprotective effect reducing cell death, white matter demyelination and reactive astrogliosis.

The cannabinoid WIN 55212-2 mitigates apoptosis and mitochondrial dysfunction after hypoxia ischemia.

Perinatal hypoxia-ischemia has significant mortality and morbidity due to there is still no specific treatment as a consequence of the complexities of hypoxic-ischemic pathophysiology. The aim of this work was to evaluate the effects of the cannabinoid agonist WIN 55212-2 on apoptotic cell death and mitochondrial dysfunction after perinatal asphyxia in fetal lambs. Animals were assigned to: one SHAM group and two hypoxic-ischemic groups that received a dose of 0.01 µg/kg WIN 55,212-2 (HI + WIN) or not (HI + VEH) after 60 min of partial occlusion of the umbilical cord, and sacrificed 3 h later. Different brain regions were separated for morphological studies, and the same territories were dissociated and analyzed by flow cytometry to quantify apoptosis, to determine mitochondrial integrity and transmembrane potential and to analyze intracellular calcium levels. Our results showed that WIN 55,212-2 reduced apoptotic cell death in all regions studied through the maintenance of mitochondrial integrity and functionality.

Hypoxic-ischemic injury in the immature brain--key vascular and cellular players.

Over the past decade, much has been learned about the cellular and molecular mechanisms underlying hypoxic-ischemic (H-I) injury in the preterm human brain. The pathogenesis of H-I brain injury is now understood to be multifactorial and quite complex, depending on (i) the severity, intensity and timing of asphyxia, (ii) selective ischemic vulnerability, (iii) the degree of maturity of the brain, and (iv) the characteristics of the ensuing reoxygenation/reperfusion phase. Each of these factors has differential effects on the distinct cell populations in the brain, with certain specific cell types being particularly vulnerable in the developing brain. In this review, we discuss the role of the blood vessels and the distinct cell populations, which are the mayor constitutive elements of the immature brain, in the pathophysiology of H-I lesion. The presence of fragile and poorly anastomosed blood vessels and the existence of disturbances in the blood-brain barrier alter blood flow, vascular tone and nutrient delivery. Brain cells are sensitive to the overstimulation of neurotransmitter receptors, particularly glutamate receptors, which can provoke excitotoxicity leading to the death of neurons and other cells such as astrocytes and oligodendrocyte progenitors. Microglial activation by means of excitatory amino acids and by leukocyte migration initiates the inflammatory response giving rise to an increase in regional cerebral blood flow and promoting astrocyte and oligodendrocyte injuries. A better understanding of these aspects of H-I injury will contribute to more efficient strategies for the management of the associated damage.

Cells from the inner mass of blastocyst as a source of neural derivates for differentiation studies.

Our results show that cells derived from the inner cell mass (ICM) show a clear tendency to differentiate into the neural lineage, showing both cells and structures in different degrees of differentiation. Among the experimental paradigms used to learn about neural differentiation, there have been several lines of investigation on stem cells, including embryonic stem (ES) cells isolated from the inner cell mass of embryo and also stem cells derived from embryonic carcinoma (EC). In this work, we have used a cellular line obtained from the inner cell mass of a blastocyst. The cells were cultured and after inoculated subcutaneously in syngenic mice. The neural differentiation was predominant, and could be observed both by morphological and immunohistochemical methods. It was represented by neural-tubes, neurons and glial cells, as expressed by the presence of Microtubule-associated protein-2 (MAP-2) and glial fibrilary acidic protein. Moreover, tyrosine hydroxilase positive labelling was found in neuron-like cells, which suggest the chatecolaminergic differentiation. These results show that isolation of cells from the inner mass of blastocyst represents an easy, reproducible and cheap source of neural derivates suitable for both in vivo and in vitro differentiation studies.

The intein of the Thermoplasma A-ATPase A subunit: structure, evolution and expression in E. coli.

BACKGROUND: Inteins are selfish genetic elements that excise themselves from the host protein during post translational processing, and religate the host protein with a peptide bond. In addition to this splicing activity, most reported inteins also contain an endonuclease domain that is important in intein propagation. RESULTS: The gene encoding the Thermoplasma acidophilum A-ATPase catalytic subunit A is the only one in the entire T. acidophilum genome that has been identified to contain an intein. This intein is inserted in the same position as the inteins found in the ATPase A-subunits encoding gene in Pyrococcus abyssi, P. furiosus and P. horikoshii and is found 20 amino acids upstream of the intein in the homologous vma-1 gene in Saccharomyces cerevisiae. In contrast to the other inteins in catalytic ATPase subunits, the T. acidophilum intein does not contain an endonuclease domain.T. acidophilum has different codon usage frequencies as compared to Escherichia coli. Initially, the low abundance of rare tRNAs prevented expression of the T. acidophilum A-ATPase A subunit in E. coli. Using a strain of E. coli that expresses additional tRNAs for rare codons, the T. acidophilum A-ATPase A subunit was successfully expressed in E. coli. CONCLUSIONS: Despite differences in pH and temperature between the E. coli and the T. acidophilum cytoplasms, the T. acidophilum intein retains efficient self-splicing activity when expressed in E. coli. The small intein in the Thermoplasma A-ATPase is closely related to the endonuclease containing intein in the Pyrococcus A-ATPase. Phylogenetic analyses suggest that this intein was horizontally transferred between Pyrococcus and Thermoplasma, and that the small intein has persisted in Thermoplasma apparently without homing.

Presence of four stem cell populations in monolayer cultures derived from teratocarcinoma embryoid bodies.

In the present work, flow cytometry techniques together with morphologic studies were used to perform multiparametric analyses in cell cultures derived from CE44 teratocarcinoma embryoid bodies. The intrinsic cell parameters studied by flow cytometry were size (FALS), cytoplasmic complexity (ISS) and autofluorescence, expressed as LIGFL/FALS (green fluorescence intensity on a logarithmic scale/FALS). Our results showed that CE44 teratocarcinoma yields monolayers whose cells show a marked morphological heterogeneity and can be grouped according to flow cytometric criteria into four populations that remain stable throughout the entire time of culture. Moreover, these populations showed a different immunolabelling with the differentiation markers SSEA-1, TROMA-1 and anti-vimentin.