Native homing endonucleases can target conserved genes in humans and in animal models

In recent years, both homing endonucleases (HEases) and zinc-finger nucleases (ZFNs) have been engineered and selected for the targeting of desired human loci for gene therapy. However, enzyme engineering is lengthy and expensive and the off-target effect of the manufactured endonucleases is difficult to predict. Moreover, enzymes selected to cleave a human DNA locus may not cleave the homologous locus in the genome of animal models because of sequence divergence, thus hampering attempts to assess the in vivo efficacy and safety of any engineered enzyme prior to its application in human trials. Here, we show that naturally occurring HEases can be found, that cleave desirable human targets. Some of these enzymes are also shown to cleave the homologous sequence in the genome of animal models. In addition, the distribution of off-target effects may be more predictable for native HEases. Based on our experimental observations, we present the HomeBase algorithm, database and web server that allow a high-throughput computational search and assignment of HEases for the targeting of specific loci in the human and other genomes. We validate experimentally the predicted target specificity of candidate fungal, bacterial and archaeal HEases using cell free, yeast and archaeal assay

Variability in the Effect of 5-HTTLPR on Depression in a Large European Population: The Role of Age, Symptom Profile, Type and Intensity of Life Stressors.

BACKGROUND: Although 5-HTTLPR has been shown to influence the risk of life stress-induced depression in the majority of studies, others have produced contradictory results, possibly due to weak effects and/or sample heterogeneity. METHODS: In the present study we investigated how age, type and intensity of life-stressors modulate the effect of 5-HTTLPR on depression and anxiety in a European population cohort of over 2300 subjects. Recent negative life events (RLE), childhood adversity (CHA), lifetime depression, Brief Symptoms Inventory (BSI) depression and anxiety scores were determined in each subject. Besides traditional statistical analysis we calculated Bayesian effect strength and relevance of 5-HTTLPR genotypes in specified models. RESULTS: The short (s) low expressing allele showed association with increased risk of depression related phenotypes, but all nominally significant effects would turn to non-significant after correction for multiple testing in the traditional analysis. Bayesian effect strength and relevance analysis, however, confirmed the role of 5-HTTLPR. Regarding current (BSI) and lifetime depression 5-HTTLPR-by-RLE interactions were confirmed. Main effect, with other words direct association, was supported with BSI anxiety. With more frequent RLE the prevalence or symptoms of depression increased in ss carriers. Although CHA failed to show an interaction with 5-HTTLPR, in young subjects CHA sensitized towards the depression promoting effect of even mild RLE. Furthermore, the direct association of anxiety with the s allele was driven by young (</=30) individuals. LIMITATIONS: Our study is cross-sectional and applies self-report questionnaires. CONCLUSIONS: Albeit 5-HTTLPR has only weak/moderate effects, the s allele is directly associated with anxiety and modulates development of depression in homogeneous subgroups

Critical Review of the State-of-the-Art on Lumbar Percutaneous Cement Discoplasty

Interbody fusion is the gold standard surgery to treat lumbar disc degeneration disease but can be a high-risk procedure in elderly and polymorbid patients. Percutaneous Cement Discoplasty (PCD) is a minimally invasive technique developed to treat advanced stage of disc degeneration exhibiting a vacuum phenomenon. A patient-specificstand-alone spacer is created by filling the disc with polymethylmethacrylate cement, allowing to recover the disc height and improve the patient’s conditions. As it has recently been introduced in the lumbar spine, this review aims to present a transversal state-of-the-art of the surgery from its clinical practice and outcome to biomechanical and engineering topics. The literature was searched across multiple databases using predefined keywords over no limited period of time. Papers about vertebroplasty were excluded. Among 466 identified papers, the relevant ones included twelve clinical papers reporting the variations of the surgical technique, follow-up and complications, four papers reporting biomechanical ex vivo and numerical tests, and four letters related to published clinical papers. Papers presenting the operative practice are reported, as well as follow-ups up to four years. The papers found, consistently reported that PCD significantly improved the clinical status of the patients and maintained it after two years. Spine alignment was impacted by PCD: the sacral slope was significantly reduced, and disc height increased. The foramen opening correlated to the volume of injected cement. Substitutes to the acrylic cement exhibited better osteointegration and mechanical properties closer to bone tissue. Finally, limitations and risks of the surgery are discussed as well as potential improvements such as the development of new filling materials with better mechanical properties and biological integration or the investigation of the inner disc

MOBILIDADES, ESPAÇOS E RELAÇÕES SOCIAIS: UMA BREVE ANÁLISE DO FILME “UP IN THE AIR”

Este trabalho é fruto das discussões realizadas em uma das sessões de Cineclube organizadas pelo Grupo de pesquisa “Turismo e Cultura” (UFF) no ano de 2010, dos debates ocorridos em 2011 na disciplina “Turismos, Migrações, Exílios e as Teorias Contemporâneas da Mobilidade” do curso de Doutorado em História, Política e Bens Culturais (Cpdoc/FGV) e das trocas sobrevindas da disciplina optativa que ministrei no curso de graduação em Turismo da UFF em 2011 e em 2012, intitulada “Mobilidades, Lazer e Turismo”. A partir do filme Up in the Air, este artigo pretende discutir os significados que as mobilidades, em seus mais diferentes e interdependentes modos, fornecem às relações sociais construídas nos (inter) espaços e nas redes de sociabilidade contemporâneas

Stereophotogrammetric approaches to multi-segmental kinematics of the thoracolumbar spine: a systematic review

Spine disorders are becoming more prevalent in today's ageing society. Motion abnormalities have been linked to the prevalence and recurrence of these disorders. Various protocols exist to measure thoracolumbar spine motion, but a standard multi-segmental approach is still missing. This study aims to systematically evaluate the literature on stereophotogrammetric motion analysis approaches to quantify thoracolumbar spine kinematics in terms of measurement reliability, suitability of protocols for clinical application and clinical significance of the resulting functional assessment

Decreased Openness to Experience Is Associated with Migraine-Type Headaches in Subjects with Lifetime Depression

Migraine and depression frequently occur as comorbid conditions, and it has been hypothesized that migraine with and without depression may have a different genetic background. A distinct personality trait constellation has been described in migraineurs. Less attention, however, was paid to personality differences in migraineurs with and without depression which may also shed light on differences in the neurobiological, background. The aim of our study was to investigate big five personality traits, headaches, and lifetime depression (DEP) in a large European general population sample. METHODS: Relationship between DEP, Big Five Inventory personality traits, and headaches identified by the ID-Migraine Questionnaire were investigated in 3,026 individuals from Budapest and Manchester with multivariate and logistic regression analyses. RESULTS: Both DEP and migraine(ID) showed differences in personality traits. Neuroticism was an independent risk factor for both conditions while a significant interaction effect appeared between the two in the case of openness. Namely, subjects with migraine(ID) and without DEP scored higher on openness compared to those who had depression. CONCLUSION: While we confirmed previous results that high neuroticism is a risk factor for both depression and migraine, openness to experience was significantly lower in the co-occurrence of migraine and depression. Our results suggest that increased openness, possibly manifested in optimal or advantageous cognitive processing of pain experience in migraine may decrease the risk of co-occurrence of depression and migraine and thus may provide valuable insight for newer prevention and intervention approaches in the treatment of these conditions

Assessment of foraminal decompression following discoplasty using a combination of ex vivo testing and numerical tools

: Percutaneous Cement Discoplasty (PCD) is a minimally invasive surgical technique to treat degenerated intervertebral discs. When the disc is severely degenerated, the vacuum observed in place of the nucleus pulposus can be filled with bone cement to restore the disc height, open the foramen space, and relieve pain. This study aimed to evaluate the foramen geometry change due to PCD, in the loaded spine. Cadaveric spines (n = 25) were tested in flexion and extension while Digital Image Correlation (DIC)&nbsp;measured displacements and deformations. Tests were performed on simulated pre-operative condition (nucleotomy) and after PCD. Registering DIC images and the 3D specimen geometry from CT scans, a 3D model of the specimens aligned in the experimental pose was obtained for nucleotomy and PCD. Foramen space volume was geometrically measured for both conditions. The volume of cement injected was measured to explore correlation with the change of foramen space. PCD induced a significant overall foraminal decompression in both flexion (foramen space increased by 835 ± 1289&nbsp;mm3, p = 0.001) and extension (1205 ± 1106&nbsp;mm3, p &lt; 0.001), confirming that the expected improvements of PCD show also during spine motion. Furthermore, in extension when the foramen is the most challenged, the impact of PCD on the foramen correlated with the injected cement volume

Accessible or not? That is the question! Analyzing the accessibility of the historic touristic city center of Paraty (RJ)

Paraty is a city in Rio de Janeiro, known for its natural, historical and cultural heritage, which has been recently inscribed on the World Heritage List as a Mixed Site, but it seems that it does not allow all kinds of people to visit it safely and autonomously when it comes to accessibility. Thus, this article was aimed to investigate the barriers to mobility in the Historical Center and its surroundings, specially of disabled people, as well as to analyse if there is a paradox between heritage and accessibility. From a qualitative, exploratory and descriptive research, which took place in 2018 and 2019: some techniques of Ethnographic Based Methodology were used, such as direct observations of researchers in field work, semi-structured interviews conducted with some representatives from the National Historic and Artistic Heritage Institute (IPHAN), from the Tourism and Urbanism Departments of Paraty, and the monitoring of the tourist experience of a disabled person in the Historical Center (participant observation). Furthermore, there have been examined documents, federal and municipal legislation about the accessibility in built environment and in cultural heritage sites. The results showed that there are different types of barriers in the city, though there is no legal prohibition to implement accessibility in heritage sites. It was concluded that in Paraty there is lack of effective actions by the government to reduce or extinguish several obstacles which impede full physical and spatial mobility of disabled people, mainly in the Historical Center and its surroundings.Paraty é um município do Rio de Janeiro, conhecido por seus patrimônios naturais, históricos e culturais que, recentemente, lhe renderam o título de Patrimônio Mundial na categoria Sítio Misto, apesar de parecer não possibilitar que todas as pessoas possam visitá-lo de forma autônoma e segura em termos de acessibilidade. A investigação que originou este artigo teve como objetivos reconhecer os obstáculos que prejudicam a mobilidade no Centro Histórico (CH) de Paraty e arredores, em especial de pessoas com deficiência (PCD), bem como analisar se há um paradoxo entre tombamento de patrimônio e acessibilidade. A partir de uma pesquisa qualitativo-exploratória e descritiva, foram adotados os seguintes procedimentos de base etnográfica, realizados entre 2018 e 2019: observações diretas dos pesquisadores em campo; entrevistas a partir de roteiros semiestruturados com representantes do Instituto do Patrimônio Histórico e Artístico Nacional (IPHAN) e das Secretarias de Turismo e de Urbanismo de Paraty; e o acompanhamento da experiência turística in loco de uma pessoa com deficiência física (método da observação participante). Além disso, foram examinados documentos, legislações federais e municipais que tratam da acessibilidade em patrimônios culturais. Os resultados mostraram que há muitos tipos de barreiras naquela cidade, embora não exista impedimento legal para a implementação da acessibilidade em bens imóveis e sítios históricos tombados. Portanto, conclui-se que no caso de Paraty, há ausência de ações efetivas do poder público para reduzir ou extinguir os diversos obstáculos que impedem a mobilidade físico-espacial plena das PCD, sobretudo no CH e adjacências.Paraty es una ciudad del Rio de Janeiro, reconocida por su patrimonio natural, histórico y cultural, que le ha dado, recientemente, el título de Patrimonio Mundial como Sitio Mixto, aunque parece que ella no permite que todas las personas la visiten de forma autónoma y segura, en términos de accesibilidad. Por lo tanto, el estudio que condujo este artículo tuvo como objetivos mapear las barreras que obstaculizan la movilidad en el Centro Histórico de Paraty y sus alrededores, en especial de las personas con discapacidad (PcD), así como analizar si hay una paradoja entre el patrimonio y la accesibilidad. A partir de una investigación cualitativa, exploratória y descriptiva, algunos procedimientos basados en la etnografía, ocurridos en 2018 y 2019 fueron usados: observaciones directas de los investigadores en campo, entrevistas, con un guión semiestructurado, con representantes del Instituto Nacional del Patrimonio Histórico y Artístico (IPHAN), de las Secretarías de Turismo y Urbanismo de Paraty, y también el monitoreo de la experiencia turística de una persona con discapacidad física en el Centro Histórico y cercanías (observación participante). Además, se examinaron documentos, legislación federal y municipal sobre la accesibilidad en patrimonios culturales. Los resultados mostraron que hay diferentes tipos de barreras en la ciudad, aunque no hay impedimento legal para implementar la accesibilidad en bienes inmuebles y en un sitio que es patrimonio. La conclusión es que en Paraty hay la ausencia de acciones efectivas por parte del poder público para reducir o extinguir los diversos obstáculos que impiden la movilidad físico-espacial total de las personas con discapacidad, sobre todo en Centro Histórico de Paraty y sus alrededores

A bifactor structural model of the Hungarian Pain Catastrophizing Scale and latent classes of a clinical sample

Pain catastrophizing is an exaggerated cognitive-affective response to actual or anticipated pain, usually measured by the Pain Catastrophizing Scale (PCS). Our study aimed to test the bifactor measurement model of the Hungarian PCS and to identify a catastrophizing risk group with a clinically meaningful cut-off score. The data of 404 chronic spine-related (neck, back and low-back) pain patients (mean age: 58.61 (SD = 14.34)) were used in our cross-sectional study. Besides pain-related and demographic data, pain catastrophizing and depressive symptoms were measured with questionnaires. Confirmatory factor analyses confirmed that the bifactor model outperformed the other tested measurement models, and the general catastrophizing factor was responsible for 81.5% of the explained variance. Using latent class analysis, we found that even moderately elevated pain catastrophizing score was related to more depressive symptoms and higher perceived pain intensity, and 22 score could be used as a cut-off score. Our results support the concept of global pain catastrophizing and the validity of the Hungarian PCS. Further studies are needed to evaluate the bifactor structure of this scale and the predictive value of the proposed cut-off score

Gene expression analysis indicates CB1 receptor upregulation in the hippocampus and neurotoxic effects in the frontal cortex 3 weeks after single-dose MDMA administration in Dark Agouti rats.

BACKGROUND: 3,4-methylenedioxymethamphetamine (MDMA, "ecstasy") is a widely used recreational drug known to impair cognitive functions on the long-run. Both hippocampal and frontal cortical regions have well established roles in behavior, memory formation and other cognitive tasks and damage of these regions is associated with altered behavior and cognitive functions, impairments frequently described in heavy MDMA users. The aim of this study was to examine the hippocampus, frontal cortex and dorsal raphe of Dark Agouti rats with gene expression arrays (Illumina RatRef bead arrays) looking for possible mechanisms and new candidates contributing to the effects of a single dose of MDMA (15 mg/kg) 3 weeks earlier. RESULTS: The number of differentially expressed genes in the hippocampus, frontal cortex and the dorsal raphe were 481, 155, and 15, respectively. Gene set enrichment analysis of the microarray data revealed reduced expression of 'memory' and 'cognition', 'dendrite development' and 'regulation of synaptic plasticity' gene sets in the hippocampus, parallel to the upregulation of the CB1 cannabinoid- and Epha4, Epha5, Epha6 ephrin receptors. Downregulated gene sets in the frontal cortex were related to protein synthesis, chromatin organization, transmembrane transport processes, while 'dendrite development', 'regulation of synaptic plasticity' and 'positive regulation of synapse assembly' gene sets were upregulated. Changes in the dorsal raphe region were mild and in most cases not significant. CONCLUSION: The present data raise the possibility of new synapse formation/synaptic reorganization in the frontal cortex three weeks after a single neurotoxic dose of MDMA. In contrast, a prolonged depression of new neurite formation in the hippocampus is suggested by the data, which underlines the particular vulnerability of this brain region after the drug treatment. Finally, our results also suggest the substantial contribution of CB1 receptor and endocannabinoid mediated pathways in the hippocampal impairments. Taken together the present study provides evidence for the participation of new molecular candidates in the long-term effects of MDMA