Design and synthesis of novel probes for TSPO, featuring an isoquinolinecarboxamide or quinazolinecarboxamide scaffold

TSPO is an evolutionarily well-conserved 18 kDa protein consisting of 169 amino acids,1 which is mainly located at the contact sites between the outer and inner mitochondrial membranes, although it is also expressed at low levels in other subcellular compartments, such as plasma membranes and nuclear fraction of cells.2 TSPO is involved in a variety of biological processes including cholesterol transport, steroidogenesis, calcium homeostasis, lipid metabolism, mitochondrial oxidation, cell growth and differentiation, apoptosis induction, and regulation of immune functions. TSPO is widely expressed throughout the body, with higher levels in steroid producing tissues, but also in other peripheral tissues including liver, heart, kidney, lung, immune system. TSPO is altered in different pathological conditions. Indeed it is up-regulated in various form of brain injury and inflammation, in neuropathologies and in cancer. Furthermore a decrease in TSPO levels has been found in generalized anxiety and mood disorders. Consequently, TSPO has been suggested as a promising diagnostic marker for the evaluation of the state and progression of related diseases. [11C]PK11195 was the first tracer to be consistently used in PET imaging studies of neuroinflammation, although numerous limitations have been pointed out, including a high level of non specific binding, and a poor signal-to-noise ratios which complicate its quantification.3 Subsequently, many groups worldwide were actively engaged in a search for TSPO ligands with improved capacities to quantify TSPO expression. In this thesis work it has been prepared the sulfoxide precursor of [18F](R)-PK14105. The isoquinolinecarboxamide, PK14105, has been previously labelled with fluorine-18 (t1/2 – 109.7 min) and evaluated in rat as a prospective radiotracer for imaging human TSPO 18 kDa with PET. This radiotracer initially appeared promising, but extensive evaluation was hindered by its rather difficult radiosynthesis based on substitution of a chloro-substituent with cyclotron–produced [18F]fluoride ion. Recently, it has been shown that simple p-nitroarylsulfoxides react readily with [18F]fluoride ion to produce the corresponding [18F]p-nitrofluoroarenes in good radiochemical yields. 1. Gavish, M.; Bachman, I.; Shoukrun, R.; Katz, Y.; Veenman, L.; Weisinger, G.; Weizman. Pharmacol. Rev. 1999, 51, 619-640. 2. Papadopoulos, V.; Baraldi, M.; Guilarte, T.R.; Knudsen, T.B.; Lacapère, J.; Lindemann, P.; Norenberg, M.D.; Nutt, D.; Weizman, A.; Zhang, M.-R.; Gavish, M. Trends Pharmacol. Sci. 2006, 27, 402-409. 3. Venneti, S.; Wiley, C.A. Prog. Neurobiol. 2006, 80, 308-322

La haine envers la femme en tant que mobile dans oreste d’euripide

En la tragedia Orestes de Eurípides se representan los sucesos acontecidos en la ciudad de Argos luego del asesinato de Clitemnestra. El protagonista de la obra es el matricidaOrestes, a quien se lo muestra sumamente excitado en la planificación de un nuevocrimen, el de Helena y su hija Hermíone. En el presente trabajo nos proponemos analizar el odio a la mujer como el móvil de la trama desde la teoría psicoanalítica de orientación lacaniana. El desprecio se constituye como un antecedente de la acción trágica al trasladarse de Clitemnestra hacia Helena, figura emblemática de la mitología griega por ser considerada la mujer más hermosa.In Euripides’ tragedy Orestes are represented the events occurred in the city of Argos after the murder of Clytemnestra. The main character of the play is the matricide Orestes, who is displayed extremely excited in the planning of a new crime, the one of Helena and her daughter Hermione. This paper aims to analyze hatred towards the woman as motivation of the plot from the psychoanalytic theory of Lacanian orientation. Contempt becomes an antecedent of the tragic action by moving from Clytemnestra towards Helena, emblematic figure of Greek mythology for being considered as the most beautiful woman.La tragédie Oreste d’Euripide présente les événements qui ont eu lieu à Argos après le meurtre de Clytemnestre. Le protagoniste de l’œuvre est le matricide Oreste, qui est dépeint comme quelqu’un de très excité planifiant un nouveau crime, celui d’Hélène et de sa fille Hermione. Ce texte a pour but d’analyser la haine envers la femme en tant que mobile de la trame sous l’angle de la théorie psychanalytique à orientation lacanienne. Le mépris apparaît comme antécédent de l’action tragique puisqu’il passe de Clytemnestre à Hélène, figure emblématique de la mythologie grecque car considérée comme la plus belle femme.Fil: Perczyk, Cecilia Josefina. Universidad Nacional de Hurlingham; Argentina. Universidad de Buenos Aires. Facultad de Psicología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Lombardi, Gabriel. Universidad de Buenos Aires. Facultad de Psicología; Argentin

Detection of possible factors favouring the evolution of migraine without aura into chronic migraine

In a minority of cases, the natural history of migraine without aura (MO) is characterised over time by its evolution into a form of chronic migraine (CM). In order to detect the possible factors predicting this negative evolution of MO, we searched in our Headache Centre files for all clinical records that met the following criteria: (a) first visit between 1976 and 1998; (b) diagnosis of MO or of common migraine at the first observation, with or without association with other primary headache types; (c) <15 days per month of migraine at the first observation; and (d) at least one follow-up visit at least 10 years after the first visit. The patients thus identified were then divided into two groups based on a favourable/steady evolution (Group A: n = 243, 195 women and 48 men) or an unfavourable evolution (Group B: n = 72, 62 women and 10 men) of their migraine over time. In the two groups, we compared various clinical parameters that were present at the first observation or emerged at the subsequent follow-up visits. The parameters that were statistically significantly more frequent in Group B--and can therefore be considered possible negative prognostic factors--were: (a) ≥ 10 days per month of migraine at the first observation; (b) presence of depression at the first visit in males; and (c) onset of depression or arterial hypertension after the first observation but before transformation to CM in females. Based on these findings, in MO patients the high frequency of migraine attacks, comorbidity with depression, and the tendency to develop arterial hypertension should require particular attention and careful management to prevent evolution into CM

Morphology of the interstitial cells of rat polycystic ovaries: an experimental study

PURPOSES: To evaluate the histomorphometry of ovarian interstitial cells, as well as the blood sex steroid concentrations of female rats with polycystic ovaries induced by continuous light. METHODS: Twenty female rats were divided into two groups: Control Group - in the estrous phase (CtrlG), and a group of rats with polycystic ovaries induced by continuous illumination (POG). CtrlG animals were maintained on a light period from 07:00 a.m. to 07:00 p.m., and POG animals with continuous illumination (400 Lux) for 60 days. After this period all animals were anesthetized and blood was collected for the determination of serum estradiol (E2), progesterone (P4), and testosterone (T), followed by removal of the ovaries that were fixed in 10% formalin and processed for paraffin embedding. Five-µm histological sections were stained with hematoxylin and eosin and used for histomorphometric analysis. Morphological analyses, cyst count, determination of concentration and of the nuclear volume of interstitial cells were performed with the aid of a light microscope adapted to a high resolution camera (AxioCam), whose images were transmitted to and analyzed by the computer using AxioVision Rel 4.8 software (Carl Zeiss). Data were analyzed statistically by the Student's t-test (pCtrlG=73.2±6.5, pCtrlG=80.6±3.9, pPOG=4.2±1.5, pCtrlG=63.6±16.5, pCtrlG=6.9±3.2, pGCtrl=73,2±6,5; pGCtrl=80,6±3,9; pGOP=4,2±1,5; pGCtrl=63,6±16,5; pGCtrl=6,9±3,2; p<0,05) em relação aos animais do GCtrl. CONCLUSÃO: As células intersticiais do ovário policístico da rata provavelmente provêm dos cistos ovarianos devido degeneração das células da granulosa e diferenciação das células da teca interna. As elevações dos níveis séricos de testosterona e de estradiol provavelmente provêm do aumento significativo da atividade celular e da área ocupada pelas células intersticiais.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Departamento de Morfologia e GenéticaUniversidade de São Paulo Faculdade de Medicina Departamento de Obstetrícia e GinecologiaUNIFESP, EPM, Depto. de Morfologia e GenéticaSciEL

Charcot-Marie-Tooth Type 2B: A New Phenotype Associated with a Novel RAB7A Mutation and Inhibited EGFR Degradation

The rare autosomal dominant Charcot-Marie-Tooth type 2B (CMT2B) is associated with mutations in the RAB7A gene, involved in the late endocytic pathway. CMT2B is characterized by predominant sensory loss, ulceromutilating features, with lesser-to-absent motor deficits. We characterized clinically and genetically a family harboring a novel pathogenic RAB7A variant and performed structural and functional analysis of the mutant protein. A 39-year-old woman presented with early-onset walking diculties, progressive distal muscle wasting and weakness in lower limbs and only mild sensory signs. Electrophysiology demonstrated an axonal sensorimotor neuropathy. Nerve biopsy showed a chronic axonal neuropathy with moderate loss of all caliber myelinated fibers. Next-generation sequencing (NGS) technology revealed in the proband and in her similarly affected father the novel c.377A&gt;G (p.K126R) heterozygous variant predicted to be deleterious. The mutation affects the biochemical properties of RAB7 GTPase, causes altered interaction with peripherin, and inhibition of neurite outgrowth, as for previously reported CMT2B mutants. However, it also shows differences, particularly in the epidermal growth factor receptor degradation process. Altogether, our findings indicate that this RAB7A variant is pathogenic and widens the phenotypic spectrum of CMT2B to include predominantly motor CMT2. Alteration of the receptor degradation process might explain the different clinical presentations in this family

Deletion of Porcn in Mice Leads to Multiple Developmental Defects and Models Human Focal Dermal Hypoplasia (Goltz Syndrome)

Focal Dermal Hypoplasia (FDH) is a genetic disorder characterized by developmental defects in skin, skeleton and ectodermal appendages. FDH is caused by dominant loss-of-function mutations in X-linked PORCN. PORCN orthologues in Drosophila and mice encode endoplasmic reticulum proteins required for secretion and function of Wnt proteins. Wnt proteins play important roles in embryo development, tissue homeostasis and stem cell maintenance. Since features of FDH overlap with those seen in mouse Wnt pathway mutants, FDH likely results from defective Wnt signaling but molecular mechanisms by which inactivation of PORCN affects Wnt signaling and manifestations of FDH remain to be elucidated.We introduced intronic loxP sites and a neomycin gene in the mouse Porcn locus for conditional inactivation. Porcn-ex3-7flox mice have no apparent developmental defects, but chimeric mice retaining the neomycin gene (Porcn-ex3-7Neo-flox) have limb, skin, and urogenital abnormalities. Conditional Porcn inactivation by EIIa-driven or Hprt-driven Cre recombinase results in increased early embryonic lethality. Mesenchyme-specific Prx-Cre-driven inactivation of Porcn produces FDH-like limb defects, while ectodermal Krt14-Cre-driven inactivation produces thin skin, alopecia, and abnormal dentition. Furthermore, cell-based assays confirm that human PORCN mutations reduce WNT3A secretion.These data indicate that Porcn inactivation in the mouse produces a model for human FDH and that phenotypic features result from defective WNT signaling in ectodermal- and mesenchymal-derived structures

Susceptibility of Human Lymphoid Tissue Cultured ex vivo to Xenotropic Murine Leukemia Virus-Related Virus (XMRV) Infection

BACKGROUND: Xenotropic murine leukemia virus-related virus (XMRV) was generated after a recombination event between two endogenous murine leukemia viruses during the production of a prostate cancer cell line. Although the associations of the XMRV infection with human diseases appear unlikely, the XMRV is a retrovirus of undefined pathogenic potential, able to replicate in human cells in vitro. Since recent studies using animal models for infection have yielded conflicting results, we set out an ex vivo model for XMRV infection of human tonsillar tissue to determine whether XMRV produced by 22Rv1 cells is able to replicate in human lymphoid organs. Tonsil blocks were infected and infection kinetics and its pathogenic effects were monitored RESULTS: XMRV, though restricted by APOBEC, enters and integrates into the tissue cells. The infection did not result in changes of T or B-cells, immune activation, nor inflammatory chemokines. Infectious viruses could be recovered from supernatants of infected tonsils by reinfecting DERSE XMRV indicator cell line, although these supernatants could not establish a new infection in fresh tonsil culture, indicating that in our model, the viral replication is controlled by innate antiviral restriction factors. CONCLUSIONS: Overall, the replication-competent retrovirus XMRV, present in a high number of laboratories, is able to infect human lymphoid tissue and produce infectious viruses, even though they were unable to establish a new infection in fresh tonsillar tissue. Hereby, laboratories working with cell lines producing XMRV should have knowledge and understanding of the potential biological biohazardous risks of this virus

Mass testing of the JUNO experiment 20-inch PMTs readout electronics

The Jiangmen Underground Neutrino Observatory (JUNO) is a multi-purpose, large size, liquid scintillator experiment under construction in China. JUNO will perform leading measurements detecting neutrinos from different sources (reactor, terrestrial and astrophysical neutrinos) covering a wide energy range (from 200 keV to several GeV). This paper focuses on the design and development of a test protocol for the 20-inch PMT underwater readout electronics, performed in parallel to the mass production line. In a time period of about ten months, a total number of 6950 electronic boards were tested with an acceptance yield of 99.1%