Vagus Nerve Stimulation (VNS) Modulates Synaptic Plasticity in the Infralimbic Cortex via Trk-B Receptor Activation to Reduce Drug-Seeking in Male Rats.

Drugs of abuse cause changes in the prefrontal cortex (PFC) and associated regions that impair inhibitory control over drug-seeking. Breaking the contingencies between drug-associated cues and the delivery of the reward during extinction learning reduces relapse. Vagus nerve stimulation (VNS) has previously been shown to enhance extinction learning and reduce drug-seeking. Here we determined the effects of VNS-mediated release of brain-derived neurotrophic factor (BDNF) on extinction and cue-induced reinstatement in male rats trained to self-administer cocaine. Pairing 10 d of extinction training with VNS facilitated extinction and reduced drug-seeking behavior during reinstatement. Rats that received a single extinction session with VNS showed elevated BDNF levels in the medial PFC as determined via an enzyme-linked immunosorbent assay. Systemic blockade of tropomyosin receptor kinase B (TrkB) receptors during extinction, via the TrkB antagonist ANA-12, decreased the effects of VNS on extinction and reinstatement. Whole-cell recordings in brain slices showed that cocaine self-administration induced alterations in the ratio of AMPA and NMDA receptor-mediated currents in Layer 5 pyramidal neurons of the infralimbic cortex (IL). Pairing extinction with VNS reversed cocaine-induced changes in glutamatergic transmission by enhancing AMPAR currents, and this effect was blocked by ANA-12. Our study suggests that VNS consolidates the extinction of drug-seeking behavior by reversing drug-induced changes in synaptic AMPA receptors in the IL, and this effect is abolished by blocking TrkB receptors during extinction, highlighting a potential mechanism for the therapeutic effects of VNS in addiction.

Vagus nerve stimulation (VNS) modulates synaptic plasticity in the rat infralimbic cortex via Trk-B receptor activation to reduce drug-seeking.

Drugs of abuse cause changes in the prefrontal cortex (PFC) and associated regions that impair inhibitory control over drug-seeking. Breaking the contingencies between drug-associated cues and the delivery of the reward during extinction learning reduces relapse. Vagus nerve stimulation (VNS) has previously been shown to enhance extinction learning and reduce drug-seeking. Here we determined the effects of VNS-mediated release of brain-derived neurotrophic factor (BDNF) on extinction and cue-induced reinstatement in rats trained to self-administer cocaine. Pairing 10 days of extinction training with VNS facilitated extinction and reduced drug-seeking behavior during reinstatement. Rats that received a single extinction session with VNS showed elevated BDNF levels in the medial PFC as determined via an enzyme-linked immunosorbent assay (ELISA). Systemic blockade of Tropomyosin receptor kinase B (TrkB) receptors during extinction, via the TrkB antagonist ANA-12, decreased the effects of VNS on extinction and reinstatement. Whole-cell recordings in brain slices showed that cocaine self-administration induced alterations in the ratio of AMPA and NMDA receptor-mediated currents in layer 5 pyramidal neurons of the infralimbic cortex (IL). Pairing extinction with VNS reversed cocaine-induced changes in glutamatergic transmission by enhancing AMPAR currents, and this effect was blocked by ANA-12. Our study suggests that VNS consolidates extinction of drug-seeking behavior by reversing drug-induced changes in synaptic AMPA receptors in the IL, and this effect is abolished by blocking TrkB receptors during extinction, highlighting a potential mechanism for the therapeutic effects of VNS in addiction.

Phenanthrene Degradation by Photosynthetic Bacterial Consortium Dominated by Fischerella sp.

Microbial degradation of aromatic hydrocarbons is an emerging technology, and it is well recognized for its economic methods, efficiency, and safety; however, its exploration is still scarce and greater emphasis on cyanobacteria-bacterial mutualistic interactions is needed. We evaluated and characterized the phenanthrene biodegradation capacity of consortium dominated by Fischerella sp. under holoxenic conditions with aerobic heterotrophic bacteria and their molecular identification through 16S rRNA Illumina sequencing. Results indicated that our microbial consortium can degrade up to 92% of phenanthrene in five days. Bioinformatic analyses revealed that consortium was dominated by Fischerella sp., however different members of Nostocaceae and Weeksellaceae, as well as several other bacteria, such as Chryseobacterium, and Porphyrobacter, were found to be putatively involved in the biological degradation of phenanthrene. This work contributes to a better understanding of biodegradation of phenanthrene by cyanobacteria and identify the microbial diversity related.

Some phenomena of the cap-pushing response in honey bees (Apis mellifera spp.).

The cap-pushing response (CPR) is a new free-flying technique used to study learning and memory in honey bees. Bees fly to a target where they push a cap to reveal a hidden food source. When combined with traditional odor and color targets, the CPR technique opens the door to additional choice preference tests in honey bees. To facilitate the use of the CPR technique, three experiments were conducted. Experiment 1 investigates the impact of extended training on the CPR response and its role in extinction. Experiment 2 explores the role of CPR in overshadowing, and Experiment 3 explores the effects of electric shock punishment on the CPR technique. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

FishShapes v1: Functionally relevant measurements of teleost shape and size on three dimensions.

Teleost fishes account for 96% of all fish species and exhibit a spectacular variety of body forms. Teleost lineages range from deep bodied to elongate (e.g., eels, needlefish), laterally compressed (e.g., ribbonfish) to globular (e.g., pufferfish), and include uniquely shaped lineages such as seahorses, flatfishes, and ocean sunfishes. Adaptive body shape convergence within fishes has long been hypothesized but the nature of the relationships between fish form and ecological and environmental variables remain largely unknown at the macroevolutionary scale. To facilitate the investigation of the interacting factors influencing teleost body shape evolution we measured eight functionally relevant linear traits on adult-sized specimens along with specimen mass. Linear measurements of standard length, maximum body depth, maximum fish width, lower jaw length, mouth width, head depth, minimum caudal peduncle depth, and minimum caudal peduncle width were taken in millimeters with calipers, or tape measures for oversized specimens. We measured these traits on a total of 16,523 specimens (1-3 specimens per species) at the Smithsonian National Museum of Natural History and took approximately 7000 person hours of data collection to complete. The data went through a three-step error-checking process to clean and validate the data and then species averages were calculated. We present the complete specimen data set, which encompasses approximately one-fifth of extant teleost species diversity, spanning ~90% of teleost families and ~96% of orders. The species and family names are compatible with the taxonomy used by FishBase and the order information with the phylogenetically informed taxonomy of Betancur-R and colleagues published in 2014. This dataset is licensed under Creative Commons CC0 1.0 Universal (CC0 1.0) but please cite this paper when using the data or a subset of it.

Intervenciones de prevención sobre el consumo de alcohol en jóvenes universitarios / Prevention interventions of alcohol consumption in young university students

Introducción: La ingesta de alcohol está condicionada por aspectos individuales y culturales. Objetivo: Identificar el efecto de intervenciones realizadas en el contexto latinoamericano sobre pautas de consumo o factores de riesgo asociados al consumo de alcohol en jóvenes universitarios. Materiales y métodos: Revisión sistemática a partir de la pregunta PICO, Se realizó búsqueda desde abril a agosto del 2020 en las bases de PubMed, CUIDEN, BVS, Scielo, Google Scholar y Repositorios Gubernamentales. Se utilizaron descriptores DeCS y MeSH, en español, inglés y portugués con los operadores AND y OR. Criterios de elegibilidad: estudios experimentales y cuasi experimentales publicados entre 2014 y 2020. Se obtuvieron 49 artículos, la lectura crítica permitió seleccionar 8 a los cuales se les aplicaron las escalas AMSTAR2, TREND y CONSORT quedando 6 artículos para análisis. Según la Resolución 008430/93, Artículo 10, se consideró como investigación sin riesgo. Resultados: Intervenciones unicomponente reportaron efectos sobre creencias conductuales, actitudes, conocimiento de la sustancia, rendimiento académico, menor frecuencia de consumo y no conducir bajo efectos del alcohol. Las intervenciones multicomponente disminuyen en 3.03% el riesgo de consumo y reportan percepción positiva respecto a la utilidad de las actividades desarrolladas, satisfacción de expectativas, satisfacción general, calidad de materiales empleados, asistencia y puntualidad. Discusión: Los estudios sugieren implementar nuevas intervenciones, estrategias y políticas en salud primaria para promover una transformación social, educativa y sanitaria que generen un impacto favorable para mitigar la problemática actual de consumo de alcohol en jóvenes universitarios. Conclusión: Esta revisión sistemática permitió sintetizar y valorar la evidencia disponible frente a intervenciones unicomponente y multicomponente realizadas en el contexto latinoamericano y del caribe que modifican positivamente factores de riesgo y pautas de consumo en jóvenes universitarios.

Introduction: Individual and cultural aspects condition alcohol intake. Objetive:To identify the effect of interventions conducted in Latin America on consumption patterns or associated risk factors for alcohol consumption among young university students. Materials and Methods:Systematic review based on a PICO question. A literature search was conducted from April to August 2020 in PubMed, CUIDEN, BVS, Scielo, Google Scholar, and Governmental Repositories. DeCS and MeSH descriptors in Spanish, English, and Portuguese were used with Boolean operators AND and OR. Eligibility criteria included experimental and quasi-experimental studies published between 2014 and 2020. Forty-nine articles were identified, and critical reading allowed the selection of 8 articles to which the AMSTAR2, TREND, and CONSORT appraisal tools were used, leaving 6 articles for analysis. In accordance with Article 10 of Resolution 008430/93, this was considered risk-free research. Results: Single-component interventions reported effects on behavioral beliefs, attitudes, knowledge of the substance, academic performance, lower frequency of drinking, and not driving under the influence of alcohol. Multicomponent interventions reduced consumption risk by 3.03% and reported positive perceptions regarding the usefulness of activities, expectations fulfillment, overall satisfaction, quality of materials used, attendance, and punctuality. Discussion:The studies suggest implementing new interventions, strategies and policies in primary health to promote a social, educational and health transformation that generate a favorable impact to mitigate the current problem of alcohol consumption in university students. Conclusions: This systematic review made it possible to synthesize and evaluate the available evidence on single- and multicomponent interventions in Latin America and the Caribbean that positively modify risk factors and consumption patterns in young university students.

Introdução: A ingestão de álcool é condicionada por aspectos individuais e culturais. Objetivo: Identificar o efeito de intervenções realizadas no contexto latino-americano sobre os padrões de consumo ou fatores de risco associados ao consumo de álcool em estudantes universitários. Materiais e métodos: Revisão sistemática com base na pergunta PICO, realizada de abril a agosto de 2020 nas bases de dados PubMed, CUIDEN, BVS, Scielo, Google Scholar e Government Repositories. Foram utilizados os descritores DeCS e MeSH, em espanhol, inglês e português com operadores AND e OR. Critérios de elegibilidade: estudos experimentais e quase-experimentais publicados entre 2014 e 2020. Obtiveram-se 49 artigos, a leitura crítica permitiu selecionar 8, aos quais foram aplicadas as escalas AMSTAR2, TREND e CONSORT, restando 6 artigos para análise. De acordo com a Resolução 008430/93, artigo 10, foi considerada pesquisa livre de risco. Resultados: Intervenções unicomponentes relataram efeitos sobre crenças comportamentais, atitudes, conhecimento da substância, desempenho acadêmico, menor frequência de consumo e não dirigir sob efeito de álcool. As intervenções multicomponentes reduzem o risco de consumo em 3,03% e relatam uma percepção positiva quanto à utilidade das atividades realizadas, satisfação das expectativas, satisfação geral, qualidade dos materiais utilizados, assiduidade e pontualidade. Discussão: Os estudos sugerem a implementação de novas intervenções, estratégias e políticas na atenção primária à saúde para promover uma transformação social, educacional e sanitária que gere impacto favorável para mitigar o problema atual do consumo de álcool em estudantes universitários. Conclusões: Esta revisão sistemática permitiu sintetizar e avaliar as evidências disponíveis sobre intervenções unicomponentes e multicomponentes realizadas no contexto latino-americano e caribenho que modificam positivamente os fatores de risco e os padrões de consumo em estudantes universitários.

SynapseJ: An Automated, Synapse Identification Macro for ImageJ.

While electron microscopy represents the gold standard for detection of synapses, a number of limitations prevent its broad applicability. A key method for detecting synapses is immunostaining for markers of pre- and post-synaptic proteins, which can infer a synapse based upon the apposition of the two markers. While immunostaining and imaging techniques have improved to allow for identification of synapses in tissue, analysis and identification of these appositions are not facile, and there has been a lack of tools to accurately identify these appositions. Here, we delineate a macro that uses open-source and freely available ImageJ or FIJI for analysis of multichannel, z-stack confocal images. With use of a high magnification with a high NA objective, we outline two methods to identify puncta in either sparsely or densely labeled images. Puncta from each channel are used to eliminate non-apposed puncta and are subsequently linked with their cognate from the other channel. These methods are applied to analysis of a pre-synaptic marker, bassoon, with two different post-synaptic markers, gephyrin and N-methyl-d-aspartate (NMDA) receptor subunit 1 (NR1). Using gephyrin as an inhibitory, post-synaptic scaffolding protein, we identify inhibitory synapses in basolateral amygdala, central amygdala, arcuate and the ventromedial hypothalamus. Systematic variation of the settings identify the parameters most critical for this analysis. Identification of specifically overlapping puncta allows for correlation of morphometry data between each channel. Finally, we extend the analysis to only examine puncta overlapping with a cytoplasmic marker of specific cell types, a distinct advantage beyond electron microscopy. Bassoon puncta are restricted to virally transduced, pedunculopontine tegmental nucleus (PPN) axons expressing yellow fluorescent protein. NR1 puncta are restricted to tyrosine hydroxylase labeled dopaminergic neurons of the substantia nigra pars compacta (SNc). The macro identifies bassoon-NR1 overlap throughout the image, or those only restricted to the PPN-SNc connections. Thus, we have extended the available analysis tools that can be used to study synapses in situ. Our analysis code is freely available and open-source allowing for further innovation.

Regulation of Oxalate Metabolism in Spinach Revealed by RNA-Seq-Based Transcriptomic Analysis.

Although spinach (Spinacia oleracea L.) is considered to be one of the most nutrient-rich leafy vegetables, it is also a potent accumulator of anti-nutritional oxalate. Reducing oxalate content would increase the nutritional value of spinach by enhancing the dietary bioavailability of calcium and other minerals. This study aimed to investigate the proposed hypothesis that a complex network of genes associated with intrinsic metabolic and physiological processes regulates oxalate homeostasis in spinach. Transcriptomic (RNA-Seq) analysis of the leaf and root tissues of two spinach genotypes with contrasting oxalate phenotypes was performed under normal physiological conditions. A total of 2308 leaf- and 1686 root-specific differentially expressed genes (DEGs) were identified in the high-oxalate spinach genotype. Gene Ontology (GO) analysis of DEGs identified molecular functions associated with various enzymatic activities, while KEGG pathway analysis revealed enrichment of the metabolic and secondary metabolite pathways. The expression profiles of genes associated with distinct physiological processes suggested that the glyoxylate cycle, ascorbate degradation, and photorespiratory pathway may collectively regulate oxalate in spinach. The data support the idea that isocitrate lyase (ICL), ascorbate catabolism-related genes, and acyl-activating enzyme 3 (AAE3) all play roles in oxalate homeostasis in spinach. The findings from this study provide the foundation for novel insights into oxalate metabolism in spinach.

Microplastic pollution on sandy beaches of Puerto Rico.

Caribbean islands, including Puerto Rico, are biodiversity hotspots threatened by microplastics (<5 mm). Little is known about the extent of microplastic pollution in coastal sandy beaches of Puerto Rico. Sand from six northern beaches was collected in the high tide line to determine microplastic abundance (0.3-4.75 mm). Península La Esperanza, the most polluted beach, exhibited higher average abundance (17 items/kg dw) and diversity. High urbanization, industrial/port activities, and riverine input are likely sources of plastic debris on this beach. The other beaches showed lower and similar average abundance (3 to 7 items/kg dw) despite having distinct potential point and non-point sources. Overall, fibers (40%), fragments (28%) and foams (27%) predominated (n = 102 particles). Results showed comparable levels to other world beaches, some classified as highly contaminated, but only when transforming units to items/m2. Preliminary ATR-FTIR analysis identified mainly polyethylene. It is imperative to have plastics source reduction through waste management.

Passive Sampling of Persistent Organic Pollutants in Four Coastal Aquatic Systems of Puerto Rico: A Pilot Study.

Little is known about the presence and effects of polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) in Puerto Rico's waters. Four coastal aquatic systems were investigated using low-density polyethylene passive sampling for PCBs and OCPs in water and its overlying air. The highest total freely dissolved and gaseous concentrations of PCBs were found in Guánica Bay, with 4000 pg/L and 270 pg/m3, respectively. Five OCPs were detected, mainly in water, with greatest concentrations (pg/L) in Guánica Bay: α-HCH (7400), p,p'-DDE (390), aldrin (2000), dieldrin (420), and endrin (77). The compound α-HCH was also measured at elevated water concentrations in Condado Lagoon (5700 pg/L) and Laguna Grande (2900 pg/L). Jobos Bay did not show values of concern for these persistence organic pollutants. Levels of PCBs and OCPs in water, particularly in Guánica Bay, exceeded USEPA ambient water quality criteria values representing a human health risk regarding consumption of aquatic organisms.

Conocimiento, experimentación y uso habitual de las terapias complementarias en función del nivel de estudios en la ciudad de Burgos / Knowledge, experience and regular use of complementary therapies based on the level of education in the city of Burgos

Objetivo: describir las terapias complementarias (TC) que la población refiere como más conocidas, experimentadas ocasionalmente o llevadas a cabo de manera habitual, y analizar la relación de su uso con el nivel de estudios de los burgaleses. Método: se realizó un estudio descriptivo trasversal usando un muestreo no probabilístico por cuotas en función del nivel de educación (n= 278). Se administró un cuestionario de 13 preguntas diseñado ad hoc y previamente pilotado, que incluyó variables sociodemográficas y relacionadas con el grado de experimentación de terapias complementarias (conocimiento, experimentación ocasional o utilización habitual). Se realizaron índices descriptivos y análisis bivariantes. Resultados: se obtuvieron 293 cuestionarios válidos. Las TC más conocidas y experimentadas fueron yoga, acupuntura y homeopatía. Un 39,9% de los burgaleses era usuario habitual. Se encontraron diferencias estadísticamente significativas en el uso habitual de TC en función del nivel de estudios, tanto en el dato global como para los grupos de terapia "sistemas integrales complejos", "prácticas de manipulación y basadas en el cuerpo" , "técnicas de la mente y el cuerpo", siendo este mayor entre quienes tienen estudios superiores (p< 0,05). Conclusiones: las TC se llevan a cabo en menor proporción de lo que se conocen; sin embargo, debido a la posible repercusión sobre la salud, se recomienda diseñar intervenciones educativas dirigidas a la población para fomentar la confianza en los cuidados basados en evidencias

Objective: to describe those complementary therapies (CTs) reported by the population as more widely known, occasionally experienced, or regularly used, and to analyze the association between their use and the level of education of the Burgos population. Method: a cross-sectional descriptive study was conducted, using non-probability quota sampling, based on level of education (n= 278). A 13-question questionnaire was designed ad hoc and administered; there was a previous pilot test, and it included sociodemographical variables and those associated with the level of experience with complementary therapies (knowledge, occasional experience or regular use). Descriptive indexes and bivariate analysis were conducted. Results: in total, 293 valid questionnaires were obtained. The most widely known and experienced CTs were yoga, acupuncture and homeopathy; 39.9% of the Burgos population were regular users. Statistically significant differences were found regarding the regular use of CT based on level of education, both in the overall data and for the following therapy groups: "complex comprehensive systems", "handling and body-based practices", "mind and body techniques"; this was higher among those with higher education (p< 0.05). Conclusions: CTs are experienced in a lower proportion than they are known; however, due to their potential impact on health, it is recommended to design educational interventions targeted to the population, in order to promote trust in evidence-based care

Systematized biosynthesis and catabolism regulate citrulline accumulation in watermelon.

Citrulline, a non-protein amino acid, is present in large amounts in watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai Cucurbitaceae) fruits. Amino acid profiling of various tissues of cv. Charleston Gray during plant development confirmed progressive accumulation of citrulline only in the fruit flesh and rind tissues. Citrulline content was positively correlated with precursor (ornithine) and by-product (arginine) amino acids during fruit ripening. Genetic variation in the partitioning of citrulline and related amino acids in the flesh and rind tissues was confirmed in a sub-set of watermelon cultivars. No correlation was established between morphological fruit traits (size and rind properties) and citrulline content. To understand the regulation of citrulline accumulation, we investigated the expression of genes associated with its biosynthesis and catabolism in flesh and rind tissues during fruit development. The expression of ornithine carbamoyltransferase (OTC) involved in the ultimate step of citrulline synthesis remained steady in both tissues. The expression of N-acetylornithine aminotransferase (N-AOA) involved in the production of N-acetylornithine and N-acetylornithine deacetylase (AOD-3) involved in ornithine synthesis coincided with increasing accumulation of citrulline in flesh and rind tissues during fruit development. Down-regulation N-acetylornithine-glutamate acetyltransferase (N-AOGA) suggests the subordinate role of the non-cyclic pathway in citrulline synthesis. Eccentricity between citrulline accumulation and expression of carbamoyl phosphate synthases (CPS-1, CPS-2) during fruit development suggest that the localized synthesis of carbamoyl phosphates may not be required for citrulline synthesis. Most genes involved in citrulline break-down (Argininosuccinate synthases - ASS-1, ASS-2, and ASS-3, Argininosuccinate lyases - ASL-1, Ornithine decarboxylase - ODC, Arginine decarboxylase - ADC) were consistently down-regulated during fruit development.

The Promise and Pitfalls of Using Crowdsourcing in Research Prioritization for Back Pain: Cross-Sectional Surveys.

BACKGROUND: The involvement of patients in research better aligns evidence generation to the gaps that patients themselves face when making decisions about health care. However, obtaining patients' perspectives is challenging. Amazon's Mechanical Turk (MTurk) has gained popularity over the past decade as a crowdsourcing platform to reach large numbers of individuals to perform tasks for a small reward for the respondent, at small cost to the investigator. The appropriateness of such crowdsourcing methods in medical research has yet to be clarified. OBJECTIVE: The goals of this study were to (1) understand how those on MTurk who screen positive for back pain prioritize research topics compared with those who screen negative for back pain, and (2) determine the qualitative differences in open-ended comments between groups. METHODS: We conducted cross-sectional surveys on MTurk to assess participants' back pain and allow them to prioritize research topics. We paid respondents US $0.10 to complete the 24-point Roland Morris Disability Questionnaire (RMDQ) to categorize participants as those "with back pain" and those "without back pain," then offered both those with (RMDQ score ≥7) and those without back pain (RMDQ <7) an opportunity to rank their top 5 (of 18) research topics for an additional US $0.75. We compared demographic information and research priorities between the 2 groups and performed qualitative analyses on free-text commentary that participants provided. RESULTS: We conducted 2 screening waves. We first screened 2189 individuals for back pain over 33 days and invited 480 (21.93%) who screened positive to complete the prioritization, of whom 350 (72.9% of eligible) did. We later screened 664 individuals over 7 days and invited 474 (71.4%) without back pain to complete the prioritization, of whom 397 (83.7% of eligible) did. Those with back pain who prioritized were comparable with those without in terms of age, education, marital status, and employment. The group with back pain had a higher proportion of women (234, 67.2% vs 229, 57.8%, P=.02). The groups' rank lists of research priorities were highly correlated: Spearman correlation coefficient was .88 when considering topics ranked in the top 5. The 2 groups agreed on 4 of the top 5 and 9 of the top 10 research priorities. CONCLUSIONS: Crowdsourcing platforms such as MTurk support efforts to efficiently reach large groups of individuals to obtain input on research activities. In the context of back pain, a prevalent and easily understood condition, the rank list of those with back pain was highly correlated with that of those without back pain. However, subtle differences in the content and quality of free-text comments suggest supplemental efforts may be needed to augment the reach of crowdsourcing in obtaining perspectives from patients, especially from specific populations.

In Vivo Evaluation of a Pneumatic Extracorporeal Ventricular Assist Device for up to 90 Day Support.

In a previous study, we showed that the Vitalmex Extracorporeal Ventricular Assist Device-Pneumatic (EVAD-P)-a low-cost, pneumatically actuated, pulsatile blood pump-is easy to implant and safe for short-term in vivo support (30 ± 5 days). In the current study, we included additional 30 day experiments and assessed the safety and durability of the EVAD-P for up to 90 days of support. Using the same surgical procedure as in the previous study, we implanted the device into 14 healthy sheep. Group I subjects (n = 7) were evaluated for up to 30 days, and group II (n = 2) and group III (n = 5) subjects were evaluated for up to 49 and 93 days, respectively. After a system redesign, two of the five sheep in group III reached the scheduled end-point without device-related problems at a fixed beat rate of 56 bpm, a stroke volume (SV) of 58.0 ± 2.3 ml, and a flow of 3.5 ± 0.2 L/min. This study shows that the EVAD-P can provide safe pulsatile mechanical circulatory support (MCS) for up to 93 days. To further confirm that the system can consistently provide MCS for this duration, additional studies are recommended.

Altered responsiveness of BNST and amygdala neurons in trauma-induced anxiety.

A highly conserved network of brain structures regulates the expression of fear and anxiety in mammals. Many of these structures display abnormal activity levels in post-traumatic stress disorder (PTSD). However, some of them, like the bed nucleus of the stria terminalis (BNST) and amygdala, are comprised of several small sub-regions or nuclei that cannot be resolved with human neuroimaging techniques. Therefore, we used a well-characterized rat model of PTSD to compare neuronal properties in resilient vs PTSD-like rats using patch recordings obtained from different BNST and amygdala regions in vitro. In this model, a persistent state of extreme anxiety is induced in a subset of susceptible rats following predatory threat. Previous animal studies have revealed that the central amygdala (CeA) and BNST are differentially involved in the genesis of fear and anxiety-like states, respectively. Consistent with these earlier findings, we found that between resilient and PTSD-like rats were marked differences in the synaptic responsiveness of neurons in different sectors of BNST and CeA, but whose polarity was region specific. In light of prior data about the role of these regions, our results suggest that control of fear/anxiety expression is altered in PTSD-like rats such that the influence of CeA is minimized whereas that of BNST is enhanced. A model of the amygdalo-BNST interactions supporting the PTSD-like state is proposed.

Serum biomarkers in uncontrolled no heart-beating donors may identify kidneys that will never work after transplantation.

BACKGROUND/AIMS: Kidneys from uncontrolled non heart-beating donors achieve a good level of renal function after transplantation. However, a number of them will never function in the recipient. Our aim was to determine if serum biomarkers associated with platelet activity, inflammation and the nitric oxide system in uncontrolled non heart-beating donors may help to predict no renal function recovery after renal transplantation. METHODS: Serum levels of interleukin (IL)-6, IL-10, intercellular cell adhesion molecule-1 (ICAM-1), cyclic guanosine monophosphate (cGMP), nitrite + nitrate and platelet factor-4 (PF4) were measured using enzyme-linked immunosorbent assay (ELISA) kits in 88 uncontrolled non heart-beating donors divided according to the renal functionality achieved in the recipients into functional (n = 76) and non functional (n = 12). RESULTS: Kidneys from donors with higher IL-6 levels (>900 pg/ml) were functional after transplantation. Serum cGMP levels below 372.3 fmol/l were also associated with kidneys that recovered the renal function. However, serum levels of PF4 showed the best correlation with recovery of renal functional in the recipients since they were significantly lower in the donors whose kidneys functioned after transplantation. CONCLUSIONS: Serum PF4 levels in uncontrolled non heart-beating donors may be a good predictor for kidneys that never will reach functional recovery. Some serum cGMP, IL-6 and IL-10 levels may simply help identify kidneys that will function after transplantation.

Direct differentiation of adult ocular progenitors into striatal dopaminergic neurons.

Parkinson's disease, characterized by motor dysfunction due to the loss of nigrostriatal dopaminergic neurons, is one of the most prevalent age-related neurodegenerative disorders. Given there is no current cure, the stem cell approach has emerged as a viable therapeutic option to replace the dopaminergic neurons that are progressively lost to the disease. The success of the approach is likely to depend upon accessible, renewable, immune compatible, and non-tumorigenic sources of neural progenitors from which stable dopaminergic neurons can be generated efficaciously. Here, we demonstrate that neural progenitors derived from limbus, a regenerative and accessible ocular tissue, represent a safe source of dopaminergic neurons. When the limbus-derived neural progenitors were subjected to a well-established protocol of directed differentiation under the influence of Shh and FGF8, they acquired the biochemical and functional phenotype of dopaminergic neurons that included the ability to synthesize dopamine. Their intrastriatal transplantation in the rat model of hemi-Parkinsonism was associated with a reduction in the amphetamine-induced rotation. No tumor formation was observed 6 weeks post-transplantation. Together, these observations posit limbus-derived neural progenitors as an accessible and safe source of dopaminergic neurons for a potential autologous ex-vivo stem cell approach to Parkinson's disease.

Efectos del bosentán sobre la función vascular e inflamación de pacientes diabéticos con enfermedad vascular periférica / Bosentan effects on vascular function and inflammation in diabetic patients with peripheral vascular disease

INTRODUCCIÓN: La endotelina-1 (ET-1) interviene en la regulación del tono vasomotor y remodelado vascular y en la disfunción endotelial. El bloqueante de ET-1, bosentán, podría tener un efecto beneficioso en la enfermedad arterial. OBJETIVO: Analizar la expresión de ET-1, óxido nítrico (NO) e interleucinas 6 y 10 (IL-6, IL-10), en diabéticos con arteriopatía periférica y en controles no diabéticos. Analizar el efecto de bosentán en la expresión de mediadores inflamatorios y en la liberación de NO. PACIENTES: Un total de 3 grupos de sujetos; G1: controles no diabéticos (n = 15), G2: diabéticos con claudicación (n = 15), G3: diabéticos con lesiones tróficas (n = 15). MATERIAL Y MÉTODOS: Análisis de expresión plasmática de ET-1 mediante dot-blot, de concentraciones plasmáticas de IL-6 e IL-10 (kits de ELISA). Capacidad de liberar NO mediante kit de nitratos+nitritos. RESULTADOS: Se observa un aumento de los niveles de ET-1 (G1: 172,9; G2: 277,1; G3: 367,3; p < 0,05, unidades arbitrarias) y un descenso de IL-6 en las formas más avanzadas de la enfermedad (G1: 45,8; G2: 16,4; G3: 9,8; p < 0,05 pg/ml). Bosentán elevó los niveles de IL-6 en el grupo de lesiones tróficas hasta igualarla con el grupo control. Se observó una reducción significativa de la capacidad de liberar NO por los leucocitos en G3 (G1: 16,7; G3: 12; p < 0,05 μmol/L). Este efecto se revirtió significativamente en presencia de bosentán. CONCLUSIONES: Los niveles elevados de ET-1 podrían influir en la progresión de la EAP. En la arteriopatía avanzada parece existir un descenso de la actividad inflamatoria (disminución de IL-6), lo que podría disminuir la vasodilatación (reducción de NO). Bosentán tiene un efecto antagonista sobre estos efectos, fundamentalmente en los estadios más avanzados de la enfermedad

INTRODUCTION: Endothelin-1 (ET-1) is involved in the regulation of vasomotor tone, vascular remodeling, and endothelial dysfunction. The ET-1 blocker, osentan, could have a beneficial effect on vascular disease. OBJECTIVE: To analyze the expression of ET-1, nitric oxide (NO), and interleukins 6 and 10 (IL-6, IL-10), in diabetics with peripheral arterial disease (PAD) and non-diabetic controls. To analyze the effects of bosentan on the expression of inflammatory mediators and the release of NO. PATIENTS: G1: non-diabetic controls (n = 15), G2: diabetic patients with claudication (n = 15), G3: diabetics patients with trophic lesions (n = 15). MATERIAL AND METHODS: Analysis of plasma ET-1 expression by dot-blot, plasma concentrations of IL-6 and IL-10 (ELISA kits). Ability to release NO by nitrate + nitrite kit. RESULTS: Increased levels of ET-1 (G1: 172.9; G2: 277.1; G3: 367.3, P<.05, arbitrary units) and a decrease in IL-6 (G1: 45.8; G2: 16.4; G3: 9.8; P<.05 pg/ml) are observed in the most advanced forms of disease. Bosentan increased levels of IL-6 in the group of trophic lesions when compared with the control group. A significant reduction in the ability of NO release by leukocytes in G3 (G1: 16.7; G3: 12; P<.05 μmol/L) was observed. This effect was significantly reversed in the presence of bosentan. CONCLUSIONS: Elevated levels of ET-1 may influence the progression of PAD. In advanced artery disease, there appears to be a reduction in inflammatory activity (decrease IL-6), which could reduce the vasodilation (NO reduction). Bosentan has an antagonistic effect on these effects, mainly in the more advanced stages of the disease

Metabolic differences between white and brown fat from fasting rabbits at physiological temperature.

It has been suggested that activated brown adipose tissue (BAT) shows increased glucose metabolic activity. However, less is known about metabolic activity of BAT under conditions of fasting and normal temperature. The aim of this study was to compare the possible differences in energetic metabolism between BAT and white adipose tissue (WAT) obtained from rabbits under the conditions of physiological temperature and 24 h after fasting conditions. The study was carried out on New Zealand rabbits (n=10) maintained for a period of 8 weeks at 23±2 °C. Food was removed 24 h before BAT and WAT were obtained. Protein expression levels of the glycolytic-related protein, glyceraldehyde-3-phosphate dehydrogenase, and pyruvate dehydrogenase were higher in WAT than that in BAT. The expression level of carnitine palmitoyltransferase 1 (CPT1) and CPT2, two fatty acid mitochondrial transporters, and the fatty acid ß-oxidation-related enzyme, acyl CoA dehydrogenase, was higher in BAT than in WAT. Cytosolic malate dehydrogenase expression and malate dehydrogenase activity were higher in WAT than in BAT. However, lactate dehydrogenase expression and lactate content were significantly higher in BAT than in WAT. In summary, this study for the first time, to our knowledge, has described how under fasting and normal temperature conditions rabbit BAT seems to use anaerobic metabolism to provide energetic fuel, as opposed to WAT, where the malate-aspartate shuttle and, therefore, the gluconeogenic pathway seem to be potentiated.

Direct Differentiation of Adult Ocular Progenitors into Striatal Dopaminergic Neurons

Parkinson's disease, characterized by motor dysfunction due to the loss of nigrostriatal dopaminergic neurons, is one of the most prevalent age-related neurodegenerative disorders. Given there is no current cure, the stem cell approach has emerged as a viable therapeutic option to replace the dopaminergic neurons that are progressively lost to the disease. The success of the approach is likely to depend upon accessible, renewable, immune compatible, and non-tumorigenic sources of neural progenitors from which stable dopaminergic neurons can be generated efficaciously. Here, we demonstrate that neural progenitors derived from limbus, a regenerative and accessible ocular tissue, represent a safe source of dopaminergic neurons. When the limbus-derived neural progenitors were subjected to a well-established protocol of directed differentiation under the influence of Shh and FGF8, they acquired the biochemical and functional phenotype of dopaminergic neurons that included the ability to synthesize dopamine. Their intrastriatal transplantation in the rat model of hemi-Parkinsonism was associated with a reduction in the amphetamine-induced rotation. No tumor formation was observed 6 weeks post-transplantation. Together, these observations posit limbus-derived neural progenitors as an accessible and safe source of dopaminergic neurons for a potential autologous ex-vivo stem cell approach to Parkinson's disease.