Evidence for lack of direct causality between pain and affective disturbances in a rat peripheral neuropathy model

[Epub ahead of print]Chronic pain is frequently accompanied by the manifestation of emotional disturbances and cognitive deficits. While a causality relation between pain and emotional/cognitive disturbances is generally assumed, several observations suggest a temporal dissociation and independent mechanisms. We therefore studied Sprague-Dawley rats that presented a natural resistance to pain manifestation in a neuropathy model (spared nerve injury [SNI]) and compared their performance in a battery of behavioral paradigms-anxiety, depression and fear memory-with animals that presented a pain phenotype. Afterward, we performed an extensive volumetric analysis across prefrontal, orbitofrontal and insular cortical areas. The majority of SNI animals manifested mechanical allodynia (low threshold [LT]), but 13% were similar to Sham controls (high threshold [HT]). Readouts of spontaneous hypersensivity (paw flinches) were also significantly reduced in HT and correlated with allodynia. To increase the specificity of our findings, we segregated the SNI animals in those with left (SNI-L) and right (SNI-R) lesions and the lack of association between pain and behavior still remains. Left-lesioned animals, independent of the LT or HT phenotype, presented increased anxiety-like behaviors and decreased well-being. In contrast, we found that the insular cortex (agranular division) was significantly smaller in HT than in LT. To conclude, pain and emotional disturbances observed following nerve injury are to some extent segregated phenomena. Also, HT and LT SNI presented differences in insular volumes, an area vastly implicated in pain perception, suggesting a supraspinal involvement in the manifestation of these phenotypes.This work has been funded by the European Regional Development Fund (FEDER), through the Competitiveness Factors Operational Programme (COMPETE) and the Northern Portugal Regional Operational Programme (NORTE 2020) under the Portugal 2020 Partnership Agreement (project NORTE-01-0145-FEDER-000023). It was also funded by National and International funds, through the Foundation for Science and Technology (FCT), under the scope of the projects POCI-01-0145- FEDER-007038 and PTDC/NEU-SCC/5301/2014 and by the IASP Early Career Research Grant 2015. Researchers were supported by FCT grant numbers PD/BD/114117/2015 (MRG via Inter-University Doctoral Programme in Ageing and Chronic Disease, PhDOC), SFRH/BD/109111/2015 (AMC via PhD Program in Health Sciences), SFRH/BD/52291/2013 (ME via PhDOC), SFRH/BD/89936/2012 (SB via PhD Program in Health Sciences (Applied)), PDE/BDE/113604/2015 (RM via PhD Program in Health Sciences (Applied)), PDE/BDE/113601/2015 (PSM via PhD Program in Health Sciences (Applied)) and SFRH/BPD/80118/2011 (HA). ARS integrated the Master Programme in Health Sciences of the School of Medicine, University of Minho.info:eu-repo/semantics/acceptedVersio

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear un derstanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5–7 vast areas of the tropics remain understudied.8–11 In the American tropics, Amazonia stands out as the world’s most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepre sented in biodiversity databases.13–15 To worsen this situation, human-induced modifications16,17 may elim inate pieces of the Amazon’s biodiversity puzzle before we can use them to understand how ecological com munities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple or ganism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region’s vulnerability to environmental change. 15%–18% of the most ne glected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lostinfo:eu-repo/semantics/publishedVersio

Search for dark matter produced in association with bottom or top quarks in √s = 13 TeV pp collisions with the ATLAS detector

A search for weakly interacting massive particle dark matter produced in association with bottom or top quarks is presented. Final states containing third-generation quarks and miss- ing transverse momentum are considered. The analysis uses 36.1 fb−1 of proton–proton collision data recorded by the ATLAS experiment at √s = 13 TeV in 2015 and 2016. No significant excess of events above the estimated backgrounds is observed. The results are in- terpreted in the framework of simplified models of spin-0 dark-matter mediators. For colour- neutral spin-0 mediators produced in association with top quarks and decaying into a pair of dark-matter particles, mediator masses below 50 GeV are excluded assuming a dark-matter candidate mass of 1 GeV and unitary couplings. For scalar and pseudoscalar mediators produced in association with bottom quarks, the search sets limits on the production cross- section of 300 times the predicted rate for mediators with masses between 10 and 50 GeV and assuming a dark-matter mass of 1 GeV and unitary coupling. Constraints on colour- charged scalar simplified models are also presented. Assuming a dark-matter particle mass of 35 GeV, mediator particles with mass below 1.1 TeV are excluded for couplings yielding a dark-matter relic density consistent with measurements

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear understanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5,6,7 vast areas of the tropics remain understudied.8,9,10,11 In the American tropics, Amazonia stands out as the world's most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepresented in biodiversity databases.13,14,15 To worsen this situation, human-induced modifications16,17 may eliminate pieces of the Amazon's biodiversity puzzle before we can use them to understand how ecological communities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple organism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region's vulnerability to environmental change. 15%–18% of the most neglected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lost

Leucine-Rich repeat kinase 2 (LRRK2) in Parkinson’s Disease - a role in endocytosis of synaptic vesicles

Dissertação de mestrado em Biologia Celular e Molecular. apresentada ao Departamento Ciências da Vida da Faculdade de Ciências e Tecnologia da Universidade de Coimbra.A doença de Parkinson (PD) é uma perturbação cerebral e prolongada que se caracteriza pelo inclusão de corpos de Lewy (LB) e pela degeneração dos neurónios dopaminérgicos da substância nigra pars compacta (SNpc) até ao estriado. É a segunda doença neurodegenerativa mais comum e tem como principais sintomas a dificuldade em controlar os movimentos voluntários, movimentos lentos, tremor, rigidez, instabilidade postural e demência. Actualmente, a patogênese da PD não está totalmente clarificada mas alguns estudos têm sugerido que o envelhecimento, factores ambientais e genéticos desempenham um papel preponderante para o desenvolvimento da doença. Mais recentemente, estudos genéticos têm descrito várias mutações no gene Leucine- Rich Repeat Kinase 2 (LRRK2) que causam PD do tipo autossómico dominante com início tardio sendo clinicamente impossível diferenciar da PD idiopática. De facto, as mutações no gene LRRK2 são responsáveis por 5-10% dos casos familiares e 1-2% dos casos esporádicos da PD. A proteína LRRK2 é uma ampla e complexa proteína com diversos domínios exibindo duas principais actividades enzimáticas, GTPase e cinase. A mutação mais conhecida, G2019S, leva ao aumento da actividade cinase, enquanto mutações no domínio GTPase, tal como R1441C/G, também afectam a actividade cinase. A função biológica/patológica da proteína LRRK2 permanece desconhecida, mas várias evidências descrevem que esta proteína desempenha papéis no encurtamento das dendrites, disfunção mitocondrial, tradução proteica, autofagia, libertação de neurotransmissores e na endocitose de vesículas sinápticas. De facto, a proteína LRRK2 tem sido descrita como uma proteína reguladora devido às suas interacções com elementos chave para o tráfego das vesículas sinápticas dentro da célula pré-sináptica tais como a proteína Rab5b, subunidades do complexo AP-2, glicoproteína 2A da vesícula sináptica (SV2A), NEM-sensitive factor (NSF) e a clathrin coat assembly protein, AP180. Neste projecto, culturas primárias de hipocampo de ratinhos Wild Type e LRRK2 Knockout foram estabelecidas para estudar, via whole-cell patch clamp, o papel da LRRK2 na reciclagem de vesículas sinápticas e na libertação de neurotransmissores. A ausência de LRRK2 e a inibição farmacológica da actividade cinase da proteína LRRK2 prejudica a libertação de neurotransmissores, ao nível da célula pré-sináptica, devido a perturbações na endocitose de vesículas sinápticas. Juntamente com estas observações, o nosso estudo mostra que existe uma conservação evolucionária desta função fisiológica em Drosophila melanogaster e em neurónios de mamíferos. Perante isto, a LRRK2 também regula, em neurónios de mamíferos, a endocitose de vesículas sinápticas na célula pré-sináptica através de um ciclo de fosforilação da endofilina.Parkinson’s disease (PD), the second most common neurodegenerative disorder, is characterized by progressive degeneration of dopaminergic projections from the substantia nigra pars compacta (SNpc) to the striatum and presence of Lewy bodies (LBs). PD affects the control of voluntary movement leading to tremor, postural imbalance, rigidity, and slowness of movement, depression and dementia. The pathogenesis of PD is not fully understood but many studies have suggested that aging, environmental factors and genetic susceptibility play an important role. Human genetics has defined several mutations in the Leucine-rich repeat kinase 2 (LRRK2) gene as a cause of late-onset autosomal dominant PD that is clinically indistinguishable from idiopathic PD. LRRK2 mutations account for 5-10% of familial and 1– 2% of sporadic PD cases. LRRK2 is a large, complex, multidomain protein displaying kinase and GTPase enzymatic activities and multiple protein–protein interaction domains. The best characterized mutation, G2019S, leads to increased kinase activity, while mutations in the GTPase domain, such as R1441C/G, have been reported to influence kinase activity as well. The biological/ pathological function of LRRK2 remains to be established, but several lines of evidence describe a role in decreased neurite outgrowth, mitochondrial dysfunction, increased protein translation, altered synaptic vesicles endocytosis and autophagy. LRRK2 has been suggested as a regulatory protein by its interaction with key elements of synaptic vesicle trafficking within the recycling pool such as Rab5b, AP-2 complex subunits, synaptic vesicle glycoprotein 2A (SV2A), NEM-sensitive factor (NSF) and clathrin coat assembly protein AP180. In this project different primary neuronal cultures of Wild Type (WT) and LRRK2 Knockout (KO) mouse were established to study, via whole-cell patch clamp recordings, the role of LRRK2 in synaptic vesicle recycling and neurotransmitter release. The absence of LRRK2 and the pharmacologic inhibition of LRRK2 kinase activity impair the neurotransmission, at level of pre-synaptic cell by disrupting the endocytosis of synaptic vesicles. Together, our work shows that there is evolutionary conservation of this previously identified physiologic function of LRRK2 from fly to mammalian neurons. Also in mammalian neurons, LRRK2 regulates the endocytosis of synaptic vesicles in the presynaptic cell, via an endophilin phosphorylation cycle

EDUCAÇÃO BRASILEIRA parâmetros contemporâneos e suas garantias

Nesses meandros projetistas da nossa sociedade dessas incipientes décadas do século XXI, passa-se a pensar a contemplação educacional e suas garantias na constituição brasileira. Esse projeto se dá a partir e com a disciplina do Projeto Integrador e, certamente, por ser um trabalho de pesquisa no ensino superior – na graduação de Direito – ancorado por uma instituição de certame pedagógico não tradicional, permite-se ressaltar a correspondência da temática para compreensões históricas e a materialização das expectativas para, pelo menos, até a metade deste vigente século. Você leitor(a) inicia esta obra atravessando a previsibilidade da educação em matéria legal na constituição brasileira. Preciso destacar como orientador e ministrante da disciplina de Projeto Integrador e coordenador do GPESC – Grupo de Pesquisa em Saberes Contemporâneos apoiado pela RECAM – Revista Camalotes, periódico científico desta instituição, que as integralidades presentes aqui são diversas e estão para além da interação entre os(as) nossos(as) acadêmicos(as). O conteúdo é caríssimo à sociedade brasileira, pois, pensar as garantias educacionais e, principalmente, as suas progressões nos faz pensar que ainda não atingimos e não atingiremos o ideal, mas certamente, caminhamos para uma contemplativa missão de corresponder ao ideal, isso nos conforta, isso nos motiva enquanto operadores(as) da pesquisa na bancada da educação, sobretudo, no ensino superior