Localization of specific erythropoietin binding sites in defined areas of the mouse brain.

The main physiological regulator of erythropoiesis is the hematopoietic growth factor erythropoietin (EPO), which is induced in response to hypoxia. Binding of EPO to the EPO receptor (EPO-R), a member of the cytokine receptor superfamily, controls the terminal maturation of red blood cells. So far, EPO has been reported to act mainly on erythroid precursor cells. However, we have detected mRNA encoding both EPO and EPO-R in mouse brain by reverse transcription-PCR. Exposure to 0.1% carbon monoxide, a procedure that causes functional anemia, resulted in a 20-fold increase of EPO mRNA in mouse brain as quantified by competitive reverse transcription-PCR, whereas the EPO-R mRNA level was not influenced by hypoxia. Binding studies on mouse brain sections revealed defined binding sites for radioiodinated EPO in distinct brain areas. The specificity of EPO binding was assessed by homologous competition with an excess of unlabeled EPO and by using two monoclonal antibodies against human EPO, one inhibitory and the other noninhibitory for binding of EPO to EPO-R. Major EPO binding sites were observed in the hippocampus, capsula interna, cortex, and midbrain areas. Functional expression of the EPO-R and hypoxic upregulation of EPO suggest a role of EPO in the brain.

Influence of different genotypes in the pattern of selenoprotein expression in response to Brazil nut supplementation

The micronutrient selenium is essential to human physiology. As the amino acid selenocysteine, it is inserted into selenoproteins with a wide range of functions including antioxidant capacity, thyroid hormone metabolism, improvement of immune system, brain function, fertility and reproduction. Low selenium status has been associated with increased risk for chronic diseases, such as cancer, type-2 diabetes and cardiovascular disease. In this context, several studies have been conducted in order to investigate if selenium supplementation could reduce the risk of such diseases. However, genetic variations may interfere in the response of individuals to a dietary intervention and must be considered as a important source of inter-individual variation. Therefore, this study was conducted was conducted to investigate the influence of genetic variations in selenoproteins genes on the response to an intervention with Brazil nuts, the richest source of selenium known in nature. The study included 130 healthy volunteers with both genders, aged 20 to 60 years old selected in University of São Paulo. They received nuts for 8 weeks, eating one nut a day, and did a washout period for more 8 weeks. All volunteers had a blood sampling collection every 4 weeks during 4 months, in a total of 5. The following analysis were done: anthropometric measurements, lipid profile, plasma malondialdehyde, plasma and erythrocyte Se, selenoprotein P, plasma and erythrocyte GPx activity, gene expression of GPX1, SEPP1, SELS and SEP15. The volunteers were also genotyped for SNPs rs1050450, rs3811699, rs1800699, rs713041, rs3877899, rs7579, rs34713741 and rs5845. Each unit of Brazil nut provided an average of 300 µg of selenium. All 130 volunteers completed the protocol. The concentrations of total cholesterol and glucose decreased after 8 weeks of supplementation. Moreover, HDL concentrations were higher for carriers of the variant T allele for GPX4_rs713041. The frequencies of the variant genotypes were 5,4% for rs1050450, rs3811699 e rs1088668, 10% for rs3877899, 19,2% for rs713041 e rs7579, 11,5% for rs5845 and 8,5% for rs34713741. The levels of the five biomarkers increased significantly after supplementation. In addition, erythrocyte GPx activity was influenced by rs1050450, rs713041 and rs5845; erythrocyte selenium was influenced by rs5845 and plasma selenium by rs3877899. Gene expression of GPX1, SEPP1 and SEP15 were higher after supplementation. The SNP rs1050450 influenced GPX1 mRNA expression and rs7579 influenced SEPP1 mRNA expression. Therefore, it can be concluded that the supplementation with one of Brazil nut for 8 weeks was efficient to reduce total cholesterol and glucose levels and to increase the concentrations of the main biomarkers of selenium status in healthy adults. Furthermore, our results suggest that GPX4_rs713041 might interfere on HDL concentrations and GPx1 activity, GPX1_rs1050450 might interfere on GPx1 activity, SEP15_rs5845 might interfere on GPx1 activity and erythrocyte selenium and SEPP1_3877899 might interfere on plasma Se levels. Therefore, the effect of genetic variations should be considered in future nutritional interventions evaluating the response to Brazil nut supplementation.

The Utility of the CVLT-C as an Embedded Validity Indicator After Pediatric Mild Traumatic Brain Injury.

Results of neuropsychological examinations depend on valid data. Whereas clinicians previously believed that clinical skill was sufficient to identify non-credible performance by examinees on standard tests, research demonstrates otherwise. Consequently, studies on measures to detect suspect effort in adults have received tremendous attention in the previous twenty years, and incorporation of validity indicators into neuropsychological examinations is now seen as integral. Few studies exist that validate methods appropriate for the measurement of effort in pediatric populations. Of extant studies, most evaluate standalone measures originally developed for use with adults. The present study examined the utility of indices from the California Verbal Learning Test – Children's Version (CVLT-C) as embedded validity indicators in a pediatric sample. Participants were 225 outpatients aged 8 to 16 years old referred for clinical assessment after mild traumatic brain injury (mTBI). Non-credible performance (n = 39) was defined as failure of the Medical Symptom Validity Test (MSVT). Logistic regression demonstrated that only the Recognition Discriminability index was predictive of MSVT failure (OR = 2.88, p < .001). A cutoff of z ≤ -1.0 was associated with sensitivity of 51% and specificity of 91%. In the current study, CVLT-C Recognition Discriminability was useful in the identification of non-credible performance in a sample of relatively high-functioning pediatric outpatients with mTBI. Thus, this index can be added to the short list of embedded validity indicators appropriate for pediatric neuropsychological assessment.

Development of the DORI-TBI: An Investigation to Develop a New Screening Measure to Determine Consultation with a School-Based Brain Injury Resource Team

Brain injury is the leading cause of disability and death in children in the United States. Student re-entry into the school setting following a traumatic brain injury is crucial to student success. Multidisciplinary teams within the school district comprised of individuals with expertise in brain injury are ideal in implementing student specific treatment plans given their specialized training and wide range of expertise addressing student needs. Therefore, the purpose of this study is to develop and initially validate a quantitative instrument that school personnel can use to determine if a student, identified as having a traumatic brain injury, will benefit from district-level consultation from a brain injury team. Three studies were designed to investigate the research questions. In study one, the planning and construction of the DORI-TBI was completed. Study two addressed the content validity of the DORI-TBI through a comparison analysis with other referral forms, content review with experts in the field of TBI, and cognitive interviews with professionals to test the usability of the new screening tool. In study three, a field administration was conducted using vignettes to measure construct validity. Results produced a valid and reliable new screening instrument that can aid school-based teams to more efficiently utilize district level consultation with a brain injury support team.

Towards Closed-loop Deep Brain Stimulation: Behavior Recognition from Human STN

Deep brain stimulation (DBS) provides significant therapeutic benefit for movement disorders such as Parkinson’s disease (PD). Current DBS devices lack real-time feedback (thus are open loop) and stimulation parameters are adjusted during scheduled visits with a clinician. A closed-loop DBS system may reduce power consumption and side effects by adjusting stimulation parameters based on patient’s behavior. Thus behavior detection is a major step in designing such systems. Various physiological signals can be used to recognize the behaviors. Subthalamic Nucleus (STN) Local field Potential (LFP) is a great candidate signal for the neural feedback, because it can be recorded from the stimulation lead and does not require additional sensors. This thesis proposes novel detection and classification techniques for behavior recognition based on deep brain LFP. Behavior detection from such signals is the vital step in developing the next generation of closed-loop DBS devices. LFP recordings from 13 subjects are utilized in this study to design and evaluate our method. Recordings were performed during the surgery and the subjects were asked to perform various behavioral tasks. Various techniques are used understand how the behaviors modulate the STN. One method studies the time-frequency patterns in the STN LFP during the tasks. Another method measures the temporal inter-hemispheric connectivity of the STN as well as the connectivity between STN and Pre-frontal Cortex (PFC). Experimental results demonstrate that different behaviors create different m odulation patterns in STN and it’s connectivity. We use these patterns as features to classify behaviors. A method for single trial recognition of the patient’s current task is proposed. This method uses wavelet coefficients as features and support vector machine (SVM) as the classifier for recognition of a selection of behaviors: speech, motor, and random. The proposed method is 82.4% accurate for the binary classification and 73.2% for classifying three tasks. As the next step, a practical behavior detection method which asynchronously detects behaviors is proposed. This method does not use any priori knowledge of behavior onsets and is capable of asynchronously detect the finger movements of PD patients. Our study indicates that there is a motor-modulated inter-hemispheric connectivity between LFP signals recorded bilaterally from STN. We utilize a non-linear regression method to measure this inter-hemispheric connectivity and to detect the finger movements. Our experimental results using STN LFP recorded from eight patients with PD demonstrate this is a promising approach for behavior detection and developing novel closed-loop DBS systems.

Sampling scarab beetles in tropical forests: the effect of light source and night sampling periods

Light traps have been used widely to sample insect abundance and diversity, but their performance for sampling scarab beetles in tropical forests based on light source type and sampling hours throughout the night has not been evaluated. The efficiency of mercury-vapour lamps, cool white light and ultraviolet light sources in attracting Dynastinae, Melolonthinae and Rutelinae scarab beetles, and the most adequate period of the night to carry out the sampling was tested in different forest areas of Costa Rica. Our results showed that light source wavelengths and hours of sampling influenced scarab beetle catches. No significant differences were observed in trap performance between the ultraviolet light and mercury-vapour traps, whereas these two methods caught significantly more species richness and abundance than cool white light traps. Species composition also varied between methods. Large differences appear between catches in the sampling period, with the first five hours of the night being more effective than the last five hours. Because of their high efficiency and logistic advantages, we recommend ultraviolet light traps deployed during the first hours of the night as the best sampling method for biodiversity studies of those scarab beetles in tropical forests.

The Benefits of Sampling Sports During Childhood

Sampling may promote prolonged engagement in sport by limiting physical injuries (Fraser-Thomas et al., 2005). Overtraining injuries are a concern for young athletes who specialize in one sport and engage in high volumes of deliberate practice (Hollander, Meyers, & Leunes, 1995; Law, Côté, & Ericsson, 2007). For instance, young gymnasts who practice for over 16 hours a week have been shown to have higher incidences of back injuries (Goldstein, Berger, Windier, & Jackson, 1991). A sampling approach in child-controlled play (e.g. deliberate play) rather than highly adult-controlled practice (e.g. deliberate practice) has been proposed as a strategy to limit overuse and other sport-related injuries (Micheli, Glassman, & Klein, 2000). In summary, sampling may protect against sport attrition by limiting sport related injuries and allowing children to have early experiences in sport that are enjoyable. Psychosocial Benefits of Sampling Only a small percentage of children who participate in school sports ever become elite athletes. Therefore, the psychosocial outcomes of sport participation are particularly important to consider. Recent studies with youth between the ages of 11 to 17 have found that those who are involved in a variety of extracurricular activities (e.g. sports, volunteer, arts) score more favourably on outcome measures such as Grade Point Average (GPA; Fredricks & Eccles, 2006a) and positive peer relationships (Fredricks & Eccles, 2006b) than youth who participate in fewer activities. These patterns are thought to exist due to each extracurricular activity bringing its own distinct pattern of socialization experiences that reinforce certain behaviours and/or teach various skills (Fredricks & Eccles, 2006b; Rose-Krasnor, Bussen, Willoughby, & Chambers, 2006). This contention is corroborated by studies of children and youths' experiences in extracurricular activities indicating that youth have unique experiences in each activity that contribute to their development (Hansen, Larson, & Dworkin, 2003; Larson, Hansen, & Moneta, 2006). This has led Wilkes and Côté (2007) to propose that children who sample different activities (through their own choice or by virtue of parental direction), have a greater chance of developing the following five developmental outcomes compared to children who specialize in one activity: 1) life skills, 2) prosocial behaviour, 3) healthy identity, 4) diverse peer groups and 5) social capital.

Control of neuronal signalling pathways by CYP46A1, an enzime involved in brain cholesterol homeostasis

Tese de doutoramento, Farmácia (Biologia Celular e Molecular), Universidade de Lisboa, Faculdade de Farmácia, 2016

A Three-Dimensional Stereotaxic MRI Brain Atlas of the Cichlid Fish Oreochromis mossambicus

The African cichlid Oreochromis mossambicus (Mozambique tilapia) has been used as a model system in a wide range of behavioural and neurobiological studies. The increasing number of genetic tools available for this species, together with the emerging interest in its use for neurobiological studies, increased the need for an accurate hodological mapping of the tilapia brain to supplement the available histological data. The goal of our study was to elaborate a three-dimensional, high-resolution digital atlas using magnetic resonance imaging, supported by Nissl staining. Resulting images were viewed and analysed in all orientations (transverse, sagittal, and horizontal) and manually labelled to reveal structures in the olfactory bulb, telencephalon, diencephalon, optic tectum, and cerebellum. This high resolution tilapia brain atlas is expected to become a very useful tool for neuroscientists using this fish model and will certainly expand their use in future studies regarding the central nervous system.

Cortical Abnormalities in Adults and Adolescents with Major Depression based on Brain Scans from 20 Cohorts Worldwide in the ENIGMA Major Depressive Disorder Working Group

The neuro-anatomical substrates of major depressive disorder (MDD) are still not well understood, despite many neuroimaging studies over the past few decades. Here we present the largest ever worldwide study by the ENIGMA (Enhancing Neuro Imaging Genetics through Meta-Analysis) Major Depressive Disorder Working Group on cortical structural alterations in MDD. Structural T1-weighted brain magnetic resonance imaging (MRI) scans from 2148 MDD patients and 7957 healthy controls were analysed with harmonized protocols at 20 sites around the world. To detect consistent effects of MDD and its modulators on cortical thickness and surface area estimates derived from MRI, statistical effects from sites were meta-analysed separately for adults and adolescents. Adults with MDD had thinner cortical gray matter than controls in the orbitofrontal cortex (OFC), anterior and posterior cingulate, insula and temporal lobes (Cohen’s d effect sizes: −0.10 to −0.14). These effects were most pronounced in first episode and adult-onset patients (>21 years). Compared to matched controls, adolescents with MDD had lower total surface area (but no differences in cortical thickness) and regional reductions in frontal regions (medial OFC and superior frontal gyrus) and primary and higher-order visual, somatosensory and motor areas (d: −0.26 to −0.57). The strongest effects were found in recurrent adolescent patients. This highly powered global effort to identify consistent brain abnormalities showed widespread cortical alterations in MDD patients as compared to controls and suggests that MDD may impact brain structure in a highly dynamic way, with different patterns of alterations at different stages of life.

New insights into the pharmacokinetics and pharmacodynamics of natalizumab treatment for patients with multiple sclerosis, obtained from clinical and in vitro studies.

BACKGROUND The monoclonal antibody natalizumab (NAT) inhibits the migration of lymphocytes throughout the blood-brain barrier by blocking very late antigen (VLA)-4 interactions, thereby reducing inflammatory central nervous system (CNS) activity in patients with multiple sclerosis (MS). We evaluated the effects of different NAT treatment regimens. METHODS We developed and optimised a NAT assay to measure free NAT, cell-bound NAT and VLA-4 expression levels in blood and cerebrospinal fluid (CSF) of patients using standard and prolonged treatment intervals and after the cessation of therapy. RESULTS In paired CSF and blood samples of NAT-treated MS patients, NAT concentrations in CSF were approximately 100-fold lower than those in serum. Cell-bound NAT and mean VLA-4 expression levels in CSF were comparable with those in blood. After the cessation of therapy, the kinetics of free NAT, cell-bound NAT and VLA-4 expression levels differed. Prolonged intervals greater than 4 weeks between infusions caused a gradual reduction of free and cell-bound NAT concentrations. Sera from patients with and without NAT-neutralising antibodies could be identified in a blinded assessment. The NAT-neutralising antibodies removed NAT from the cell surface in vivo and in vitro. Intercellular NAT exchange was detected in vitro. CONCLUSIONS Incorporating assays to measure free and cell-bound NAT into clinical practice can help to determine the optimal individual NAT dosing regimen for patients with MS.

Oil Shale Symposium : Sampling Analysis and Quality Assurance, March 1979 /

Mode of access: Internet.

Beryllium sampling methods: Comparison of two personal sample collection methods with the AEC sample collection method as used for one year in a beryllium production facility. /

Item 499-F-2

National Center for Statistics and Analysis collected technical studies. Volume I: accident data analysis of vehicle crashworthiness - ten papers.

Mode of access: Internet.

National Center for Statistics and Analysis collected technical studies. Volume II: accident data analysis of occupant injuries and crash characteristics - eight papers.

Mode of access: Internet.