Traffic-related air pollution, chronic stress, and changes in exhaled nitric oxide and lung function among a panel of children with asthma living in an underresourced community.

We examined associations between short-term exposure to traffic-related air pollutants (TRAP) and airway inflammation and lung function in children with asthma, and whether these associations are modified by chronic psychological stress. Residents of underresourced port-adjacent communities in New Jersey were concerned about the cumulative impacts of exposure to TRAP, particularly diesel-engine truck emissions, and stress on exacerbation of asthma among children. Children with asthma aged 9-14 (n = 35) were recruited from non-smoking households. We measured each participant's (1) continuous personal exposure to black carbon (BC, a surrogate of TRAP) at 1-min intervals, (2) 24-h integrated personal exposure to nitrogen dioxide (NO2), (3) daily fractional exhaled nitric oxide (FeNO), and (4) lung function for up to 30 consecutive days. Personal BC was recorded by micro-aethalometers. We measured daily FeNO using the NIOX MINO, forced expiratory volume in one second (FEV1), and forced vital capacity (FVC) using Easy One Frontline spirometers. Chronic stress was measured with the UCLA Life Stress Interview for Children. The association was examined using linear mixed-effect models. In the fully adjusted model, an interquartile range (IQR) increase in BC at lag 0-6 h before the FeNO measurement was associated with 8 % (95 % CI: 3 % - 12 %) increase in FeNO, whereas an IQR increase in BC at lag 7-12 h and lag 0-24 h were associated with 6 % (95 % CI: 2 % - 11 %) and 7 % (2 % - 12 %) FeNO increases, respectively. There were no significant lung function changes per IQR increase in BC. No interactions were observed between chronic stress and BC on FeNO. Chronic stress was negatively associated with individual average FeNO levels. Our findings suggest that higher levels of BC exposure within the prior 24 h increased airway inflammation levels in children with asthma, with the strongest effect observed within the first 6 h.

Thalamocortical Projections Are Significantly Impaired in the R6/2 Mouse Model of Huntington's Disease.

As Huntington's disease (HD) progresses, there is a significant loss of neurons in the striatum in addition to a distinct thinning of the cerebral cortex. Despite an early presence of sensorimotor deficits in patients with HD, electrophysiological studies designed to assess the integrity of thalamocortical circuits are sparse. Using the R6/2 mouse model of HD, we provide evidence of reduced connectivity between thalamic cells and their targeted cortical regions. Whole-cell patch clamp recordings from ventral anterolateral nucleus (VAL; motor) and ventral posteromedial nucleus (VPM; somatosensory) thalamic neurons in ex vivo brain slices of R6/2 and wild-type (WT) mice revealed that cells in both thalamic nuclei of R6/2 mice exhibited significant differences in passive and active cell membrane properties (smaller cell membrane capacitances, faster decay time constants and increased input resistances) compared with WT cells. Although only cells in the VPM of symptomatic R6/2 mice had more depolarized resting membrane potentials compared with WTs, cells in both nuclei displayed increased excitability in symptomatic, but not presymptomatic, R6/2 mice. Optical activation of VAL and VPM terminals elicited smaller magnitude current responses in cortical pyramidal neurons (CPNs) in both motor cortex (M1CTX) and somatosensory barrel cortex (BCTX) of symptomatic R6/2 mice compared with CPNs in WT mice. Furthermore, we observed a decrease in the frequency of thalamocortical excitatory quantal events in R6/2 BCTX CPNs, with no genotype-dependent differences in AMPA:NMDA response amplitude ratios. These data suggest there is a decrease in the transmission of thalamocortical information that is likely because of impaired neurotransmitter release.

[Effectiveness of peri-articular infiltration with local anesthetic and adjuvants for post-surgical pain control in total knee replacement]. / Eficacia de la infiltración periarticular con anestésico local y adyuvantes para control del dolor postquirúrgico en la artroplastía total de rodilla.

INTRODUCTION: Total knee arthroplasty (TKA) is one of the most successful orthopedic treatments, however, it has been associated with severe postsurgical pain in 30-60% of patients. We propose that infiltration of the articular capsule of the knee during surgery will decrease postsurgical pain. MATERIAL AND METHODS: Experimental, randomized, double-blind study in patients undergoing unilateral TKA between April 2018 and January 2019. Patients were divided into two groups, the first infiltration with placebo and the second with anesthetic solution and adjuvants (fentanyl, epinephrine and ketorolac). Pain was measured with the visual analog scale (VAS) at 4, 6, 8, 12, 18, 24, 36 and 48 hours postsurgical, as well as the consumption of opioid analgesics and antiemetics. RESULTS: 20 patients in each group, with a follow-up of 4 weeks. There were no significant differences in demographic characteristics between the two groups. Better control of postsurgical pain was observed in the group that received infiltration with anesthetic and adjuvant, as well as a decrease in the consumption of opioid analgesics and antiemetics. There was no difference in bleeding or in the incidence of infections between the two groups. CONCLUSION: Peri-capsular infiltration is a safe and effective method, as part of multimodal analgesia in total knee arthroplasty, as it decreases postsurgical pain, opioid and antiemetic use and does not increase postsurgical bleeding.

INTRODUCCIÓN: La artroplastía total de rodilla (ATR) es uno de los tratamientos ortopédicos más exitosos; sin embargo, se ha asociado a dolor postquirúrgico intenso en 30-60% de los pacientes. Nosotros planteamos que la infiltración de la cápsula articular de la rodilla durante la cirugía disminuirá el dolor postquirúrgico. MATERIAL Y MÉTODOS: Estudio experimental, aleatorio, doble ciego, en pacientes sometidos a ATR unilateral entre Abril de 2018 a Enero de 2019. Los pacientes fueron divididos en dos grupos, el primero infiltración con placebo y el segundo con solución anestésica y adyuvantes (fentanilo, epinefrina y ketorolaco). Se cuantificó mediante escala visual análoga (EVA) del dolor a las cuatro, seis, ocho, 12, 18, 24, 36 y 48 horas postquirúrgicas, así como del consumo de analgésicos opioides y antieméticos. RESULTADOS: Veinte pacientes en cada grupo, con un seguimiento de cuatro semanas. No hubo diferencias significativas en las características demográficas entre ambos grupos. Se observó un mejor control del dolor postquirúrgico en el grupo que recibió infiltración con anestésico y adyuvante, además de una disminución en el consumo de analgésicos opioides y antieméticos. No hubo diferencia en sangrado ni en la incidencia de infecciones entre ambos grupos. CONCLUSIÓN: La infiltración pericapsular es un método seguro y eficaz, como parte de la analgesia multimodal en la artroplastía total de rodilla, ya que disminuye el dolor postquirúrgico, el consumo de opioides y antieméticos y no incrementa el sangrado postquirúrgico.

Eficacia de la infiltración periarticular con anestésico local y adyuvantes para control del dolor postquirúrgico en la artroplastía total de rodilla / Effectiveness of peri-articular infiltration with local anesthetic and adjuvants for post-surgical pain control in total knee replacement

Resumen: Introducción: La artroplastía total de rodilla (ATR) es uno de los tratamientos ortopédicos más exitosos; sin embargo, se ha asociado a dolor postquirúrgico intenso en 30-60% de los pacientes. Nosotros planteamos que la infiltración de la cápsula articular de la rodilla durante la cirugía disminuirá el dolor postquirúrgico. Material y métodos: Estudio experimental, aleatorio, doble ciego, en pacientes sometidos a ATR unilateral entre Abril de 2018 a Enero de 2019. Los pacientes fueron divididos en dos grupos, el primero infiltración con placebo y el segundo con solución anestésica y adyuvantes (fentanilo, epinefrina y ketorolaco). Se cuantificó mediante escala visual análoga (EVA) del dolor a las cuatro, seis, ocho, 12, 18, 24, 36 y 48 horas postquirúrgicas, así como del consumo de analgésicos opioides y antieméticos. Resultados: Veinte pacientes en cada grupo, con un seguimiento de cuatro semanas. No hubo diferencias significativas en las características demográficas entre ambos grupos. Se observó un mejor control del dolor postquirúrgico en el grupo que recibió infiltración con anestésico y adyuvante, además de una disminución en el consumo de analgésicos opioides y antieméticos. No hubo diferencia en sangrado ni en la incidencia de infecciones entre ambos grupos. Conclusión: La infiltración pericapsular es un método seguro y eficaz, como parte de la analgesia multimodal en la artroplastía total de rodilla, ya que disminuye el dolor postquirúrgico, el consumo de opioides y antieméticos y no incrementa el sangrado postquirúrgico.

Abstract: Introduction: Total knee arthroplasty (TKA) is one of the most successful orthopedic treatments, however, it has been associated with severe postsurgical pain in 30-60% of patients. We propose that infiltration of the articular capsule of the knee during surgery will decrease postsurgical pain. Material and methods: Experimental, randomized, double-blind study in patients undergoing unilateral TKA between April 2018 and January 2019. Patients were divided into two groups, the first infiltration with placebo and the second with anesthetic solution and adjuvants (fentanyl, epinephrine and ketorolac). Pain was measured with the visual analog scale (VAS) at 4, 6, 8, 12, 18, 24, 36 and 48 hours postsurgical, as well as the consumption of opioid analgesics and antiemetics. Results: 20 patients in each group, with a follow-up of 4 weeks. There were no significant differences in demographic characteristics between the two groups. Better control of postsurgical pain was observed in the group that received infiltration with anesthetic and adjuvant, as well as a decrease in the consumption of opioid analgesics and antiemetics. There was no difference in bleeding or in the incidence of infections between the two groups. Conclusion: Peri-capsular infiltration is a safe and effective method, as part of multimodal analgesia in total knee arthroplasty, as it decreases postsurgical pain, opioid and antiemetic use and does not increase postsurgical bleeding.

Exposure to traffic-related air pollution and changes in exhaled nitric oxide and DNA methylation in arginase and nitric oxide synthase in children with asthma.

BACKGROUND: Traffic-related air pollution (TRAP) has been associated with increased risk of airway inflammation in children with asthma. While epigenetic changes could potentially modulate TRAP-induced inflammatory responses, few studies have assessed the temporal pattern of exposure to TRAP, epigenetic changes and inflammation in children with asthma. Our goal was to test the time-lag patterns of personal exposure to TRAP, airway inflammation (measured as fractional exhaled nitric oxide, FeNO), and DNA methylation in the promoter regions of genes involved in nitric oxide synthesis among children with asthma. METHODS: We measured personal exposure to black carbon (BC) and FeNO for up to 30 days in a panel of children with asthma. We collected 90 buccal cell samples for DNA methylation analysis from 18 children (5 per child). Methylation in promoter regions of nitric oxide synthase (NOS1, NOS2A, NOS3) and arginase (ARG1, ARG2) was assessed by bisulfite pyrosequencing. Linear-mixed effect models were used to test the associations of BC at different lag periods, percent DNA methylation at each site and FeNO level. RESULTS: Exposure to BC was positively associated with FeNO, and negatively associated with DNA methylation in NOS3. We found strongest association between FeNO and BC at lag 0-6 h while strongest associations between methylation at positions 1 and 2 in NOS3 and BC were at lag 13-24 h and lag 0-24 h, respectively. The strengths of associations were attenuated at longer lag periods. No significant associations between exposure to TRAP and methylation levels in other NOS and ARG isoforms were observed. CONCLUSIONS: Exposure to TRAP was associated with higher levels of FeNO and lower levels of DNA methylation in the promoter regions of the NOS3 gene, indicating that DNA methylation of the NOS3 gene could be an important epigenetic mechanism in physiological responses to TRAP in children with asthma.

Aided phytostabilisation reduces metal toxicity, improves soil fertility and enhances microbial activity in Cu-rich mine tailings.

(Aided) phytostabilisation has been proposed as a suitable technique to decrease the environmental risks associated with metal(loid)-enriched mine tailings. Field scale evaluations are needed for demonstrating their effectiveness in the medium- to long-term. A field trial was implemented in spring 2011 in Cu-rich mine tailings in the NW of Spain. The tailings were amended with composted municipal solid wastes and planted with Salix spp., Populus nigra L. or Agrostis capillaris L. cv. Highland. Plant growth, nutritive status and metal accumulation, and soil physico- and bio-chemical properties, were monitored over three years (four years for plant growth). The total bacterial community, α- and ß-Proteobacteria, Actinobacteria and Streptomycetaceae were studied by DGGE of 16s rDNA fragments. Compost amendment improved soil properties such as pH, CEC and fertility, and decreased soil Cu availability, leading to the establishment of a healthy vegetation cover. Both compost-amendment and plant root activity stimulated soil enzyme activities and induced important shifts in the bacterial community structure over time. The woody plant, S. viminalis, and the grassy species, A. capillaris, showed the best results in terms of plant growth and biomass production. The beneficial effects of the phytostabilisation process were maintained at least three years after treatment.

Is "intra-operating room" thromboelastometry useful in liver transplantation? A case-control study in 303 patients.

Coagulation monitoring during liver transplantation (LT) is, even today, fundamental to reduce blood loss during surgery. Thromboelastometry (TEM) is a proven technique for controlling the various parameters that influence coagulation. However, there are no studies linking "intra-operating room" TEM (orTEM) with LT outcomes. We describe a case-control study in 303 liver graft recipients analyzing variables associated with operative complications and long-term LT outcomes. The results showed that orTEM reduced the use of blood products in patients with Model for End-Stage Liver Disease scores of ≥ 21, retransplantation, and high surgical difficulty and important intraoperative bleeding. In addition, results in survival and postoperative complications were better when orTEM was used. In conclusion, we confirm that use of orTEM is associated with less use of blood products and a lower rate of complications after LT.

Pathophysiology of Huntington's disease: time-dependent alterations in synaptic and receptor function.

Huntington's disease (HD) is a progressive, fatal neurological condition caused by an expansion of CAG (glutamine) repeats in the coding region of the Huntington gene. To date, there is no cure but great strides have been made to understand pathophysiological mechanisms. In particular, genetic animal models of HD have been instrumental in elucidating the progression of behavioral and physiological alterations, which had not been possible using classic neurotoxin models. Our groups have pioneered the use of transgenic HD mice to examine the excitotoxicity hypothesis of striatal neuronal dysfunction and degeneration, as well as alterations in excitation and inhibition in striatum and cerebral cortex. In this review, we focus on synaptic and receptor alterations of striatal medium-sized spiny (MSNs) and cortical pyramidal neurons in genetic HD mouse models. We demonstrate a complex series of alterations that are region-specific and time-dependent. In particular, many changes are bidirectional depending on the degree of disease progression, that is, early vs. late, and also on the region examined. Early synaptic dysfunction is manifested by dysregulated glutamate release in striatum followed by progressive disconnection between cortex and striatum. The differential effects of altered glutamate release on MSNs originating the direct and indirect pathways is also elucidated, with the unexpected finding that cells of the direct striatal pathway are involved early in the course of the disease. In addition, we review evidence for early N-methyl-D-aspartate receptor (NMDAR) dysfunction leading to enhanced sensitivity of extrasynaptic receptors and a critical role of GluN2B subunits. Some of the alterations in late HD could be compensatory mechanisms designed to cope with early synaptic and receptor dysfunctions. The main findings indicate that HD treatments need to be designed according to the stage of disease progression and should consider regional differences.

Behaviour of enzymatic activities and root elongation in Argiudoll soils from the Argentine Humid Pampa treated with biosolids.

The incorporation of biosolids to soil is a strategy aiming at the re-location of these materials in the environment with a useful end: soil fertilization. In this work, the response of two Argiudoll soils (one with more than 100 years of agriculture and the other, a virgin one) to biosolid incorporation was studied under laboratory conditions. To measure this response, soil enzymatic biodescriptors, such as dehydrogenase and urease activities, and tests related to plant physiology (the root elongation test) were employed. The addition of the biosolid to both soils had a stimulating effect though different on each soil according to the added dose. Adjustment of the regression line for dehydrogenase activity with root elongation was positive and statistically significant (p<0.001). Results suggest that biodescriptors employed were suitable for studying the impact of amended biosolids on different soils.

Evaluation of various tests for the diagnosis of soil contamination by 2,4,5-trichlorophenol (2,4,5-TCP).

Soil response to contamination with 2,4,5-triclorophenol was studied to test the validity of the concept of Generic Reference Levels (GRL), the main criterion used to define soil contamination. Soil samples were artificially contaminated with doses between 0 and 5000 mg kg(-1) of 2,4,5-triclorophenol, and analysed by various tests. Where possible, the response of soils to the contaminant was modelled by a sigmoidal dose-response curve in order to estimate the ED(50) values. The tests provided different responses, but only microbial biomass-C and dehydrogenase and urease activities demonstrated soil deterioration in response to contamination. The results suggest that the diagnosis of soil contamination has been greatly simplified in the legislation by the provision of a single figure for each compound, and that the GRL concept could perhaps be substituted by measurement of ED(50) values, which better reflect the alteration of a soil due to the presence of a xenobiotic substance.

Behaviour of enzymatic activities and root elongation in Argiudoll soils from the Argentine Humid Pampa treated with biosolids / Comportamiento de actividades enzimáticas y elongación de raíces en suelos Argiudoles de la Pampa Húmeda, Argentina, tratados con biosólidos

The incorporation of biosolids to soil is a strategy aiming at the re-location of these materials in the environment with a useful end: soil fertilization. In this work, the response of two Argiudoll soils (one with more than 100 years of agriculture and the other, a virgin one) to biosolid incorporation was studied under laboratory conditions. To measure this response, soil enzymatic biodescriptors, such as dehydrogenase and urease activities, and tests related to plant physiology (the root elongation test) were employed. The addition of the biosolid to both soils had a stimulating effect though different on each soil according to the added dose. Adjustment of the regression line for dehydrogenase activity with root elongation was positive and statistically significant (p<0.001). Results suggest that biodescriptors employed were suitable for studying the impact of amended biosolids on different soils.

La incorporación de biosólidos al suelo es una estrategia que tiene como objetivo la reubicación de estos materiales en el ambiente con un fin útil, como es la fertilización del suelo. En este trabajo se estudió, en condiciones controladas de laboratorio, la respuesta de dos suelos Argiudoles (uno con más de 100 años de agricultura y otro virgen) frente a la perturbación físico-química y biótica que genera la incorporación de un biosólido. Para medir esta respuesta se emplearon dos biodescriptores edáficos (las actividades deshidrogenasa y ureasa) y un tercero referido a la fisiología vegetal, la prueba de elongación de raíces. La incorporación del biosólido en ambos suelos, en general no deprimió el funcionamiento de las actividades enzimáticas estudiadas; contrariamente, según la dosis aportada tuvo un efecto estimulante, aunque diferente, entre ambos suelos. El ajuste de la recta de regresión de la actividad deshidrogenasa con la elongación de las plántulas fue positivo y altamente significativo, lo que indica la complementaridad de ambos descriptores. Los resultados obtenidos sugieren que los biodescriptores empleados resultaron aptos para estudiar el impacto que produce la incorporación de biosólidos a suelos agrícolas.

Prevalence of M. tuberculosis infection and tuberculosis disease among HIV-infected people in Spain.

OBJECTIVE: To study the prevalence of Mycobacterium tuberculosis infection (MTBI) and past/current tuberculosis (TB) among human immunodeficiency virus (HIV) infected persons in Spain. DESIGN: Longitudinal study conducted between 2000 and 2003 at 10 HIV hospital-based clinics. Data were drawn from clinical records. Associations were measured using odds ratios (ORs) and their 95% confidence intervals (95%CI). RESULTS: Of the 1242 persons who met the eligibility criteria, most were male (75%), aged <40 years (75%) and unemployed (40%). HIV infection occurred through intravenous drug use (53%), heterosexual sex (29%) and sex between men (16%). In the initial evaluation, 315 subjects had evidence of MTBI: 84 (6.8%) had a history of TB, 23 (1.8%) current TB and 208 (16.8%) latent tuberculosis infection (LTBI). MTBI was associated with male sex, age 30-49 years, contact with a TB case, homelessness, poor education, and negatively with CD4 <100 cells/mm(3). Among subjects with MTBI, past/current TB was associated with retirement/disability (OR 6, 95%CI 1.6-22.5), CD4 <200 cells/mm(3) (OR 9.7, 95%CI 3.8-24.6), viral load >55,000 copies (OR 5.3, 95%CI 1.4-20.0), and negatively, with skilled work (OR 0.4, 95%CI 0.1-1.0) or administrative/managerial/professional work (OR 0.05, 95%CI 0.01-0.4). CONCLUSION: Social context has an impact on the effectiveness of HIV and TB control programmes even in industrialised countries with free access to health care.

Infantile spasm-associated microencephaly in tuberous sclerosis complex and cortical dysplasia.

OBJECTIVE: In children with and without infantile spasms, this study determined brain volumes and cell densities in epilepsy surgery patients with tuberous sclerosis complex (TSC) and cortical dysplasia with balloon cells (CD). METHODS: We compared TSC (n = 18) and CD (n = 17) patients with normal/autopsy controls (n = 20) for MRI gray and white matter volumes and neuronal nuclei (NeuN) cell densities. RESULTS: In patients without a history of infantile spasms, TSC cases showed decreased gray and white matter volumes (-16%). In cases with a history of infantile spasms, both CD (-25%) and TSC (-35%) patients showed microencephaly. This was confirmed in monozygotic twins with TSC, where the twin with a history of spasms had cerebral volumes less (-16%) than the twin without a history of seizures. Regardless of seizure history, TSC patients showed decreased NeuN cell densities in lower gray matter (-36%), whereas CD patients had increased densities in upper cortical (+52%) and white matter regions (+65%). For TSC patients, decreased lower gray matter NeuN densities correlated with reduced MRI volumes. CONCLUSIONS: Patients with tuberous sclerosis without spasms showed microencephaly associated with decreased cortical neuronal densities. In contrast, cortical dysplasia patients without spasms were normocephalic with increased cell densities. This supports the concept that tuberous sclerosis and cortical dysplasia have different pathogenetic mechanisms despite similarities in refractory epilepsy and postnatal histopathology. Furthermore, a history of infantile spasms was associated with reduced cerebral volumes in both cortical dysplasia and tuberous sclerosis patients, suggesting that spasms or their treatment may contribute to microencephaly independent of etiology.

Modification of the degradative capacity of a soil artificially contaminated with diesel.

Samples of an Ah horizon from a Cambisol under oakwood vegetation were artificially contaminated with diesel (at doses of 20, 40, 80, 160 and 400 microl g(-1)) under laboratory conditions. The presence of the contaminant caused a decrease in the microbial biomass and in phosphomonoesterase, beta-glucosidase and particularly, urease activities. In contrast, the basal respiration and mineralization of nitrogen, specifically ammonification, both increased. The microorganisms that survived in the contaminated soil showed increased metabolic activity, as a result of their use of dead microorganisms as a substrate and of a lack of competition. Although the results indicate that the properties related to the degradative capacity of the soils varied differently in response to the contamination, the use of a biochemical quality index (including different biochemical properties), which was designed for estimating the degree of deterioration of the soil, revealed a clear decrease in the biochemical quality of the diesel-contaminated soils.

Variaciones Morfométricas de la Próstata de Chinchilla laniger (MOLINA, 1982) y de la Concentración de Testosterona Plasmática Durante un Ciclo Reproductivo Anual / Morphometric Variations of Chinchilla laniger Prostate and Plasmatic Testosterone Concentration During its Annual Reproductive Cycle

Durante un año, se estudió la próstata de individuos adultos de la especie Chinchilla laniger (MOLINA, 1982), mantenidos en cautiverio, para detectar posibles variaciones estacionales en el peso glandular y corporal, índice órgano-somático (IOS), grosor de la pared y altura del epitelio glandular y concentración de testosterona plasmática. Además, se realizó un análisis histoquímico con técnicas de PAS, diastasa-PAS y Azul de Alcian pH1 y pH 2,5. Se describió una Zona A, periférica y una Zona B, central. En ambas, las células epiteliales siempre fueron negativas al Azul de Alcian. En la Zona A, se detectó la presencia de glicoproteínas neutras con intensa reacción PAS (+++) entre octubre y abril y leve a moderada (+/++) entre mayo y septiembre, y ausencia de glicógeno. La Zona B siempre fue PAS negativa. Los valores morfométricos máximos se registraron entre mayo y julio y los mínimos entre noviembre y febrero, presentando diferencias estacionales significativas (p 0,05), con excepción del peso corporal, que no mostró variaciones durante el año. La concentración de testosterona plasmática presentó los valores mínimos en enero, febrero y junio y los máximos en abril, mayo, julio y agosto. Se concluye, que la próstata de la Chinchilla laniger en cautiverio, presenta la mayor actividad secretora durante el período otoño-invierno, respondiendo positivamente a la concentración de testosterona y coincidiendo con la máxima actividad reproductiva descrita para la especie en el mismo periodo.

The prostate of adult individuals of the species Chinchilla laniger (MOLINA, 1982) kept in captivity was studied during a one year period, in order to detect possible seasonal variations in its glandular and corporal weight, organo-somatic index (OSI), wall thickness and glandular epithelium height, and in plasmatic testosterone concentration. In addition, a histochemical analysis was performed with PAS and diastase-PAS and Blue of Alcian pH 1 and pH 2.5. A periphery Zone A and a central Zone B are described. In both zones the epithelial cells always gave negative results to Blue of Alcian. The presence of neutral glicoproteins was detected in Zone A, with high PAS (+++) reaction between October and April and slight to moderate (+/++) between May and September, and absence of glycogen in all cases. The B zone gave always negative PAS. The maximum morphometric values were recorded between May and July, while the minimum values were recorded in November and February, showing significant seasonal differences (p 0.05), excepting of corporal weight, which did not show variations during the one year period. The concentration of plasmatic testosterone showed minimum values in January, February and June, and maximum values in April, May, July, and August.

Pediatric cortical dysplasia: correlations between neuroimaging, electrophysiology and location of cytomegalic neurons and balloon cells and glutamate/GABA synaptic circuits.

Seizures in cortical dysplasia (CD) could be from cytomegalic neurons and balloon cells acting as epileptic 'pacemakers', or abnormal neurotransmission. This study examined these hypotheses using in vitro electrophysiological techniques to determine intrinsic membrane properties and spontaneous glutamatergic and GABAergic synaptic activity for normal-pyramidal neurons, cytomegalic neurons and balloon cells from 67 neocortical sites originating from 43 CD patients (ages 0.2-14 years). Magnetic resonance imaging (MRI), (18)fluoro-2-deoxyglucose positron emission tomography (FDG-PET) and electrocorticography graded cortical sample sites from least to worst CD abnormality. Results found that cytomegalic neurons and balloon cells were observed more frequently in areas of severe CD compared with mild or normal CD regions as assessed by FDG-PET/MRI. Cytomegalic neurons (but not balloon cells) correlated with the worst electrocorticography scores. Electrophysiological recordings demonstrated that cytomegalic and normal-pyramidal neurons displayed similar firing properties without intrinsic bursting. By contrast, balloon cells were electrically silent. Normal-pyramidal and cytomegalic neurons displayed decreased spontaneous glutamatergic synaptic activity in areas of severe FDG-PET/MRI abnormalities compared with normal regions, while GABAergic activity was unaltered. In CD, these findings indicate that cytomegalic neurons (but not balloon cells) might contribute to epileptogenesis, but are not likely to be 'pacemaker' cells capable of spontaneous paroxysmal depolarizations. Furthermore, there was more GABA relative to glutamate synaptic neurotransmission in areas of severe CD. Thus, in CD tissue alternate mechanisms of epileptogenesis should be considered, and we suggest that GABAergic synaptic circuits interacting with cytomegalic and normal-pyramidal neurons with immature receptor properties might contribute to seizure generation.

Calcium modulates dopamine potentiation of N-methyl-D-aspartate responses: electrophysiological and imaging evidence.

In the striatum, dopamine (DA) exerts a major modulatory influence on voltage- and ligand-gated currents. Previously we have shown that DA modulates glutamatergic neurotransmission and that the direction of this modulation depends on, among other factors, the glutamate and DA receptor subtypes activated. These effects also involve DA-induced alterations in voltage-gated Ca(2+) currents. In the present experiments, the effects of Ca(2+) channel blockers on DA and D1 receptor-dependent potentiation of N-methyl-D-aspartate (NMDA) responses were examined in vitro in striatal slices using current clamp recording techniques. DA or D1 receptor agonists consistently enhanced NMDA responses. Cadmium and the more selective L-type Ca(2+) channel antagonists nifedipine and methoxyverapamil reduced the potentiation of NMDA responses by DA or D1 receptor activation. Furthermore, studies using Ca(2+) imaging with Fluo-3 in cultured cortical or dissociated striatal neurons demonstrated that DA and D1 agonists increased intracellular Ca(2+) transients induced by NMDA. These as well as previous findings indicate that in striatal neurons at least two mechanisms contribute to the enhancement of NMDA responses by DA receptor activation, facilitation of voltage-gated Ca(2+) currents and D1 receptor activation of the cAMP-protein kinase A cascade. The existence of multiple mechanisms leading to a similar outcome allows a certain degree of redundancy in the consequences of DA modulation.

Detection of Flexibacter maritimus in fish tissue using nested PCR amplification.

A nested polymerase chain reaction (PCR) system was developed for the detection of Flexibacter maritimus from fish tissue. The total procedure for the diagnosis of marine flexibacteriosis, from the point of DNA extraction to the electrophoretic analysis, can be performed in < 4 h. This was achieved by the combination of a short thermal cycling programme with a rapid DNA extraction procedure. The assay was extremely sensitive, capable of detecting as few as 75 cfu mg(-1) fish tissue. The accuracy of the nested PCR was confirmed under field conditions using tissue samples recovered during 1993-2002 from fish suffering marine flexibacteriosis. The nested PCR method proved to be efficient for the rapid and sensitive detection of F. maritimus from fish tissues and can be used for routine diagnosis of the disease caused by this pathogen.

Ionotropic glutamate receptor expression and dopaminergic modulation in the developing subthalamic nucleus of the rat: an immunohistochemical and electrophysiological analysis.

Using standard immunohistochemical procedures, we investigated the changes in the expression of ionotropic glutamate receptor (GluR) subunits, GluRl, GluR5/6/7, and NMDAR1, in the subthalamic nucleus of developing rats. The general sequence of development for each subunit was the same. At early postnatal ages, there was dense neuropil staining and cellular clustering which progressed to decreased neuropil staining and an even distribution of conspicuous cells in the later postnatal ages and in the adult. GluR5/6/7 displayed the earliest maturation, while GluR1 exhibited the slowest maturation. These morphological changes suggest a different time course for the functionality of GluR subtypes in the developing subthalamic nucleus. Correlative electrophysiological studies demonstrated functional GluRs as early as 16 days of age. All neurons tested displayed robust responses to kainate and N-methyl-D-aspartate, and these responses were modulated by dopamine.