Unbalance between Excitation and Inhibition in Phenylketonuria, a Genetic Metabolic Disease Associated with Autism

Phenylketonuria (PKU) is the most common genetic metabolic disease with a well-documented association with autism spectrum disorders. It is characterized by the deficiency of the phenylalanine hydroxylase activity, causing plasmatic hyperphenylalaninemia and variable neurological and cognitive impairments. Among the potential pathophysiological mechanisms implicated in autism spectrum disorders is the excitation/inhibition (E/I) imbalance which might result from alterations in excitatory/inhibitory synapse development, synaptic transmission and plasticity, downstream signalling pathways, and intrinsic neuronal excitability. Here, we investigated functional and molecular alterations in the prefrontal cortex (pFC) of BTBR-Pah(enu2) (ENU2) mice, the animal model of PKU. Our data show higher frequency of inhibitory transmissions and significant reduced frequency of excitatory transmissions in the PKU-affected mice in comparison to wild type. Moreover, in the pFC of ENU2 mice, we reported higher levels of the post-synaptic cell-adhesion proteins neuroligin1 and 2. Altogether, our data point toward an imbalance in the E/I neurotransmission favouring inhibition in the pFC of ENU2 mice, along with alterations of the molecular components involved in the organization of cortical synapse. In addition to being the first evidence of E/I imbalance within cortical areas of a mouse model of PKU, our study provides further evidence of E/I imbalance in animal models of pathology associated with autism spectrum disorders

Pre-analytical stability of coagulation parameters in plasma stored at room temperature

Introduction: Haemostasis testing is influenced by many pre-analytical variables, such as storage time and temperature, which can affect the stability of coagulation factors and influence the results of coagulation assays. We investigated the stability of haemostasis tests after storage of aliquoted plasma at RT, including the variability of measurement principle and reagent used for determination. Methods: Blood samples from 20 healthy volunteers were obtained, processed to PPP and aliquoted. Aliquots were stored at RT for 0 hour, 2 hours, 4 hours, 6 hours, 8 hours, 12 hours, 24 hours and 48 hours. PT, aPTT, fibrinogen, D-Dimers and coagulation factors (FII, FV, FVII, FX, FVIII, FIX, FXI, FXII) were determined by STA-R Max (R) and ACL-TOP (R). VWF:Ag and vWF:RCo were determined by AcuStar (R). Clinically relevant changes, compared to the initial measurement, were denoted as a percentage change of > 10% according to the 99% CI. Results: For both analysers, a clinically relevant change of > 10% was observed for FV after 2 hours, FVIII after 4 hours and for aPTT, FII, FVII, FX and FXII after 48 hours of storage at RT. Statistically significant, but no clinically relevant differences were observed after 48-hours storage for PT, fibrinogen and FIX. D-Dimers, FXI, vWF: Ag and vWF: RCo were found stable up to 48 hours at RT. Conclusion: Overall, compared to the limits given by the current CLSI guidelines, for most coagulation parameters investigated in this study a longer storage period could be accepted. Time intervals for FVIII and FV dosage were shorter than recommended by the CLSI guidelines. For PT determination, our findings were consistent

Quality in coagulation and haemostasis testing

The essential elements of a quality program, specifically internal quality control (IQC) and external quality assurance (EQA), should be applied to each laboratory assay performed in order to ensure test result accuracy and precision. The coagulation laboratory plays an important role in the diagnosis and treatment of individuals with bleeding or clotting (i.e., thrombotic) disorders. Test methodologies used to assess common disorders or diseases of haemostasis are reviewed as well as the clinical relevance of each assay. The preanalytical phase of testing offers the greatest opportunity for introducing result error in the haemostasis laboratory and it is therefore imperative that samples are properly collected, transported and stored. Samples for haemostasis testing should be collected in 3.2% sodium citrate at a 9:1 blood to anticoagulant ratio and maintained at room temperature until processed. Some test processes such as platelet function testing have special processing and testing requirements. For plasma-based tests, centrifugation to obtain platelet poor plasma and testing should ideally be completed within 4 hours or the plasma frozen. IQC must be performed with each assay, at appropriate levels of the analyte and at appropriate time intervals as a means for assessing ongoing assay performance. EQA, a peer group assessment process that is supplementary to IQC, offers in addition the opportunity for evaluation of long-term performance of laboratories, including comparisons with like and unlike methodologies, and often serves as an educational resource. Participation in an EQA program is often a requirement of laboratory accreditation and there are a multitude of EQA organizations that offer programs specific to haemostasis testing with international programs providing assessment of the more specialized haemostasis assays. These programs provide invaluable information on assay specific diagnostic error rate, assay precision, accuracy, sensitivity and assessment of overall assay performance. The incorporation of IQC and EQA into a laboratory program can not only assist in the assurance that testing is reliable and accurate but also improve the quality of the testing

The arterial blood supply of the temporomandibular joint: an anatomical study and clinical implications.

Purpose: The aim of this study was to analyze three-dimensional images of the arterial supply to the temporomandibular joint. Materials and Methods: Ten patients (five men and five women, mean age 36 years) without signs or symptoms of temporomandibular disorders, who underwent contrast-enhanced computed tomographic (CT) scanning with intravenous contrast, were studied. The direct volume rendering technique of CT images was used, and a data set of images to visualize the vasculature of the human temporomandibular joint in three dimensions was created. After elaboration of the data through post-processing, the arterial supply of the temporomandibular joint was studied. Results: The analysis revealed the superficial temporal artery, the anterior tympanic artery, the deep temporal artery, the auricular posterior artery, the transverse facial artery, the middle meningeal artery, and the maxillary artery with their branches as the main arterial sources for the lateral and medial temporomandibular joint. Conclusion: The direct volume rendering technique was found to be successful in the assessment of the arterial supply to the temporomandibular joint. The superficial temporal artery and maxillary artery ran along the lateral and medial sides of the condylar neck, suggesting that these arteries are at increased risk during soft-tissue procedures such as an elective arthroplasty of the temporomandibular joint

Preliminary evidence about the effects of meditation on interoceptive sensitivity and social cognition

Background: Interoception refers to the conscious perception of body signals. Mindfulness is a meditation practice that encourages individuals to focus on their internal experiences such as bodily sensations, thoughts, and emotions. In this study, we selected a behavioral measure of interoceptive sensitivity (heartbeat detection task, HBD) to compare the effect of meditation practice on interoceptive sensitivity among long term practitioners (LTP), short term meditators (STM, subjects that completed a Mindfulness-Based Stress Reduction (MBSR) program) and controls (non-meditators). All participants were examined with a battery of different tasks including mood state, executive function and social cognition tests (emotion recognition, empathy and theory of mind). Findings: Compared to controls, both meditators’ groups showed lower levels of anxiety and depression, but no improvement in executive function or social cognition performance was observed (except for lower scores compared to controls only in the personal distress dimension of empathy). More importantly, meditators’ performance did not differ from that of nonmeditators regarding cardiac interoceptive sensitivity. Conclusion: Results suggest no influence of meditation practice in cardiac interoception and in most related social cognition measures. These negative results could be partially due to the fact that awareness of heartbeat sensations is not emphasized during mindfulness/vipassana meditation and may not be the best index of the awareness supported by the practice of meditation.Fil: Melloni, Margherita. Instituto de Neurologia Cognitiva. Laboratorio de Psicologia Experimental y Neurociencia; Argentina. Universidad Favaloro. Facultad de Medicina. Instituto de Neurociencias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Sedeño, Lucas. Instituto de Neurologia Cognitiva. Laboratorio de Psicologia Experimental y Neurociencia; Argentina. Universidad Favaloro. Facultad de Medicina. Instituto de Neurociencias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Couto, Juan Blas Marcos. Instituto de Neurologia Cognitiva. Laboratorio de Psicologia Experimental y Neurociencia; Argentina. Universidad Favaloro. Facultad de Medicina. Instituto de Neurociencias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Reynoso, Martín. Universidad Favaloro. Facultad de Medicina. Instituto de Neurociencias; Argentina. Instituto de Neurologia Cognitiva. Laboratorio de Psicologia Experimental y Neurociencia; ArgentinaFil: Gelormini Lezama, Carlos. Instituto de Neurologia Cognitiva. Laboratorio de Psicologia Experimental y Neurociencia; Argentina. Universidad Favaloro. Facultad de Medicina. Instituto de Neurociencias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Favaloro, Roberto. Universidad Favaloro. Facultad de Medicina. Instituto de Neurociencias; Argentina. Instituto de Neurologia Cognitiva. Laboratorio de Psicologia Experimental y Neurociencia; ArgentinaFil: Canales Johnson, Andres. Universidad Diego Portales; ChileFil: Sigman, Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Física de Buenos Aires; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Física. Laboratorio de Neurociencia Integrativa; Argentina. Universidad Torcuato Di Tella; ArgentinaFil: Manes, Facundo Francisco. Instituto de Neurologia Cognitiva. Laboratorio de Psicologia Experimental y Neurociencia; Argentina. Universidad Favaloro. Facultad de Medicina. Instituto de Neurociencias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ibanez Barassi, Agustin Mariano. Universidad Favaloro. Facultad de Medicina. Instituto de Neurociencias; Argentina. Instituto de Neurologia Cognitiva. Laboratorio de Psicologia Experimental y Neurociencia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

The Mastery of Space in Early Modern Political Thought Giovanni Botero, the Gallery of Maps in the Vatican and the fusion of the civitas and urbs in sixteenth-century Italy

Before the sixteenth century there was a strong emphasis in Renaissance political thought on the difference between the political city (the civitas) and the physical city (the urbs). The body politic ‘floated’ above the landscape—it was not rooted in a specific territory as we understand states to be today. In 1588 the philosopher-priest Giovanni Botero argued that wealth underpinned political power; challenging humanist narratives about the corrupting forces of wealth for the civitas. These narratives were rooted in a classical ‘politics’, which saw the civitas as a morally oriented body. Botero conceived his arguments in the context of ‘reason of state’, where politics became the art of maintaining one’s ‘stato’. While arguing that wealth was the source of civitas’ power he grounded it in the economic capacity of the urbs. Physical space became an object of political power. I argue in this thesis that the Gallery of Maps in the Vatican was a potent visual exposition of these ideas—ideas which had important implications for the rise of the territorial mercantilist ‘state’

Methods and methodologies for organic synthesis

Methods and methodologies for organic synthesi

Neurolign 3 misfolding mutations and activation of the unfolded protein response

Several forms of monogenic heritable autism spectrum disorders are associated with mutations in the neuroligin genes. The autism-linked substitution R451C in neuroligin3 induces local misfolding of its extracellular domain, causing partial retention in the ER (endoplasmic reticulum) of expressing cells. We have generated a PC12 Tet-On cell model system with inducible expression of wild-type or R451C neuroligin3 to investigate whether there is activation of the UPR (unfolded protein response) as a result of misfolded protein retention. As a positive control for protein misfolding, we also expressed the mutant G221R neuroligin3, which is known to be completely retained within the ER. Our data show that overexpression of either R451C or G221R mutant proteins leads to the activation of all three signalling branches of the UPR downstream of the stress sensors ATF6 (activating transcription factor 6), IRE1 (inositol-requiring enzyme 1) and PERK [PKR (dsRNA-dependent protein kinase)-like endoplasmic reticulum kinase]. Each branch displayed different activation profiles that partially correlated with the degree of misfolding caused by each mutation. We also show that up-regulation of BiP (immunoglobulin heavy-chain-binding protein) and CHOP [C/EBP (CCAAT/enhancer-binding protein)-homologous protein] was induced by both mutant proteins but not by wild-type neuroligin3, both in proliferative cells and cells differentiated to a neuron-like phenotype. Collectively, our data show that mutant R451C neuroligin3 activates the UPR in a novel cell model system, suggesting that this cellular response may have a role in monogenic forms of autism characterized by misfolding mutations.This work was supported by: Compagnia San Paolo, Sapienza University of Rome and Pasteur Institute - Cenci Bolognetti Foundation grants to ADJ. SJM is a MRC Senior Clinical Research Fellow [MRC Ref G1002610]. This work was also supported by National Institutes of Health grants (MH092906), from the Robert Wood Johnson Foundation to the Child Health Institute of New Jersey [grant #67038] and to the Governor's Council for Medical Research and Treatment of Autism [CAUT14APL028] to D.C.This is the final version of the article. It first appeared from Portland Press via https://doi.org/http://dx.doi.org/10.1042/BJ2015027