Is MR Spectroscopy Really the Best MR-Based Method for the Evaluation of Fatty Liver in Diabetic Patients in Clinical Practice?

Objective: To investigate if magnetic resonance spectroscopy (MRS) is the best Magnetic Resonance (MR)-based method when compared to gradient-echo magnetic resonance imaging (MRI) for the detection and quantification of liver steatosis in diabetic patients in the clinical practice using liver biopsy as the reference standard, and to assess the influence of steatohepatitis and fibrosis on liver fat quantification.Methods: Institutional approval and patient consent were obtained for this prospective study. Seventy-three patients with type 2 diabetes (60 women and 13 men; mean age, 5469 years) underwent MRI and MRS at 3.0 T. the liver fat fraction was calculated from triple-and multi-echo gradient-echo sequences, and MRS data. Liver specimens were obtained in all patients. the accuracy for liver fat detection was estimated by receiver operator characteristic (ROC) analysis, and the correlation between fat quantification by imaging and histolopathology was analyzed by Spearman's correlation coefficients.Results: the prevalence of hepatic steatosis was 92%. All gradient-echo MRI and MRS findings strongly correlated with biopsy findings (triple-echo, rho = 0.819; multi-echo, rho = 0.773; MRS, rho = 0.767). Areas under the ROC curves to detect mild, moderate, and severe steatosis were: triple-echo sequences, 0.961, 0.975, and 0.962; multi-echo sequences, 0.878, 0.979, and 0.961; and MRS, 0.981, 0.980, and 0.954. the thresholds for mild, moderate, and severe steatosis were: triple-echo sequences, 4.09, 9.34, and 12.34, multi-echo sequences, 7.53, 11.75, and 15.08, and MRS, 1.71, 11.69, and 14.91. Quantification was not significantly influenced by steatohepatitis or fibrosis.Conclusions: Liver fat quantification by MR methods strongly correlates with histopathology. Due to the wide availability and easier post-processing, gradient-echo sequences may represent the best imaging method for the detection and quantification of liver fat fraction in diabetic patients in the clinical practice.D'Or Institute for Research and EducationFundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ)DOr Inst Res & Educ, Rio de Janeiro, BrazilUniv Fed Rio de Janeiro, Rio de Janeiro, BrazilUniv Estado Rio de Janeiro, Rio de Janeiro, BrazilUniv São Paulo, Inst Phys Sao Carlos, Sao Carlos, SP, BrazilUniversidade Federal de São Paulo, São Paulo, BrazilUniv Paris Diderot Sorbonne, Paris, FranceUniversidade Federal de São Paulo, São Paulo, BrazilWeb of Scienc

Expression of HMGCS2 in intestinal epithelial cells is downregulated in inflammatory bowel disease associated with endoplasmic reticulum stress.

INTRODUCTION The Unfolded Protein Response, a mechanism triggered by the cell in response to Endoplasmic reticulum stress, is linked to inflammatory responses. Our aim was to identify novel Unfolded Protein Response-mechanisms that might be involved in triggering or perpetuating the inflammatory response carried out by the Intestinal Epithelial Cells in the context of Inflammatory Bowel Disease. METHODS We analyzed the transcriptional profile of human Intestinal Epithelial Cell lines treated with an Endoplasmic Reticulum stress inducer (thapsigargin) and/or proinflammatory stimuli. Several genes were further analyzed in colonic biopsies from Ulcerative Colitis patients and healthy controls. Lastly, we generated Caco-2 cells lacking HMGCS2 by CRISPR Cas-9 and analyzed the functional implications of its absence in Intestinal Epithelial Cells. RESULTS Exposure to a TLR ligand after thapsigargin treatment resulted in a powerful synergistic modulation of gene expression, which led us to identify new genes and pathways that could be involved in inflammatory responses linked to the Unfolded Protein Response. Key differentially expressed genes in the array also exhibited transcriptional alterations in colonic biopsies from active Ulcerative Colitis patients, including NKG2D ligands and the enzyme HMGCS2. Moreover, functional studies showed altered metabolic responses and epithelial barrier integrity in HMGCS2 deficient cell lines. CONCLUSION We have identified new genes and pathways that are regulated by the Unfolded Protein Response in the context of Inflammatory Bowel Disease including HMGCS2, a gene involved in the metabolism of Short Chain Fatty Acids that may have an important role in intestinal inflammation linked to Endoplasmic Reticulum stress and the resolution of the epithelial damage.This work was supported by grants from Ministerio de Ciencia e Innovación (MCIN) from Spain [SAF2016-78711R and PID202-11794 to EM-N and FJC]; Comunidad de Madrid [B2017/BMD-3727 to EMN and FJC]; Comunidad de Madrid (REACT-UE, ANTICIPA-CM Ref. PR38/21-24) to E.M-N and HORIZON-HLTH-2022-STAYHLTH-02 under agreement No 101095679 to FJC the European Union’s Horizon 2020 research and innovation program [ERC-2016- Consolidator Grant 725091 to DS]; MCIN/AEI/10.13039/ 501100011033 [PID2019-108157RB to DS]; la Caixa Foundation (ID 100010434) [LCF/BQ/PR20/11770008 to SW]; Instituto de Salud Carlos III (ISCIII) [PI18/00348 to VE]; ISCIII [PI21/01641 to RT-R]; Spanish National Research and Development Plan, ISCIII and FEDER [PI17/02303 and PI20/01837 to SR-P]; Proyecto Desarrollo Tecnológico [DTS19/00111 to SR-P], AEI/MICIU EXPLORA Project [BIO2017-91272-EXP to SR-P]; Programa Estratégico Instituto de Biologıa y Gene ́ ́ tica Molecular (IBGM), Junta de Castilla y León (CCVC8485) [PID2019-104218RB-I00 to DB]; NIH [DK088199 to RB] and Universidad Complutense de Madrid (UCM 920631) [CT42/ 18-CT43/18 and EB15/21 to BM-A].S

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

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

Development of a digital image analysis technique for the evaluation of fibrosis stage in liver biopsies of HBV and HCV patients

As hepatites crônicas por vírus são as mais frequentes, destacando-se os vírus das hepatites B (VHB) e C (VHC). O estudo anatomopatológico da biópsia hepática é considerado o padrão ouro para avaliar com precisão a distorção arquitetural e o grau de fibrose do parênquima do fígado, importantes fatores prognósticos para os pacientes portadores de hepatites crônicas virais. Na avaliação histopatológica atual, em adição aos relatos subjetivos das alterações histológicas, escores semiquantitativos que correlacionam achados morfológicos com graus numéricos são usados, tais como os reconhecidos escores de Ishak e METAVIR. Entretanto, em todos estes sistemas há a desvantagem da subjetividade do examinador e da incorporação de alterações categóricas, sem referências às mudanças quantitativas do colágeno hepático. Técnicas de análise de imagens digitais (AID) que fornecem quantificação objetiva dos graus de fibrose em amostras histológicas têm sido desenvolvidas. Todavia, o alto custo e dificuldade ao acesso das tecnologias descritas restringem seu uso a poucos centros especializados. Este estudo visa o desenvolvimento de uma técnica de custo acessível para a análise de imagens digitais da fibrose hepática em hepatites crônicas virais. Foram estudadas 304 biópsias de pacientes com hepatite crônica por vírus B e C, obtidas através de agulhas Menghini. Todas as amostras tinham pelo menos 15 mm de comprimento ou cinco espaços-porta completos e foram coradas pelo método Tricrômico de Masson. O estadiamento foi feito por um único hepatopatologista experiente, sem o conhecimento dos dados clínicos dos pacientes. Os escores de Ishak e METAVIR foram aplicados. As imagens microscópicas foram digitalizadas. Os índices de fibrose foram determinados de forma automatizada, em técnica desenvolvida no programa Adobe Photoshop. Para o escore de Ishak, observamos os seguintes índices de Fibrose (IF) médios: 0,8% 0,0 (estágio 0), 2.4% 0,6 (estágio 1), 4,7% 1,6 (estágio 2), 7,4% 1,4 (estágio 3), 14,9% 3,7 (estágio 4), 23,4% 2,9 (estágio 5) e 34,5% 1,5 (estágio 6). Para a classificação METAVIR: 0,8% 0,1 (estágio F0), 3,8% 1,8 (estágio F1), 7,4% 1,4 (estágio F2), 20,4% 5,2 (estágio F3) e 34,5% 1,5 (estágio F4). Observamos uma excelente correlação entre os índices de fibrose da AID e os escores de Ishak (r=0,94; p<0,001) e METAVIR (r=0,92; p<0,001). Em relação à indicação de tratamento antiviral, foi observado IF médio de 16,4%. Em relação ao diagnóstico de cirrose, foi observado IF médio de 26,9%, para o escore de Ishak, e 34,5% para a classificação METAVIR. A reprodutibilidade intra-observador foi excelente. Este novo método de análise de imagens digitais para a quantificação de fibrose hepática tem custo acessível e foi desenvolvido com tecnologia que está disponível em todo o mundo, permitindo identificar com precisão todos os estágios de fibrose, com excelente reprodutibilidade intra-observador.The hepatitis caused by viruses are the most frequent, principally the virus B (HBV) and C (HCV) hepatitis. The anatomopathologic study of the liver biopsy is considered the gold standard for the evaluation of architectural distortion and degree o fibrosis of the livers parenchyma, important prognostic factors for the patients with chronic viral hepatitis. In the actual histopathological evaluation, in addition to subjective reports of histological alterations, semiquantitative scores that correlate morphological findings with numeric degrees are used, such as the renowned Ishak and METAVIR scores. However, in all of these systems there is the disadvantage of the subjectiveness of the examinator and the incorporation of categorical description of architectural changes without reference to quantitative changes in liver collagen. Digital Image Analysis techniques (DIA) that provide objective quantification of the degrees of liver fibrosis in histological samples have been developed. But the high cost and worldwide unavailability of the technologies described restrained their use to a few specialized centers. This study aims at developing an affordable technique for livers fibrosis DIA in chronic viral hepatitis. Three hundred and four core needle biopsies obtained by Menghini needles from patients with virus B and virus C hepatitis were studied. All samples had at least 15 mm in length or five complete portal tracts and were stained with the Massons Trichrome Method. The staging by Ishaks and METAVIRs scores was done by a single experienced hepatopathologist, without the knowledge patients clinical data. The microscopic images were digitalized. The fibrosis indexes were calculated in an automated way by a new technique developed in the Adobe Photoshop software. For the Ishak score the following median Fibrosis Indexes (FI) were observed: 0,8% 0,0 (stage 0), 2.4% 0,6 (stage 1), 4,7% 1,6 (stage 2), 7,4% 1.4 (stage 3), 14,9% 3.7 (stage 4), 23,4% 2.9 (stage 5) e 34,5% 1,5 (stage 6). For the METAVIR classification: 0,8% 0,1 (stage F0), 3,8% 1.8 (stage F1), 7,4% 1.4 (stage F2), 20,4% 5,2 (stage F3) e 34,5% 1,5 (stage F4). We found an excellent correlation between the DIA fibrosis indexes and the Ishaks (r=0.94; p<0.001) and METAVIR (r=0.92; p<0.001) stages. In relation to the indication for antiviral treatment, the mean FI observed was 16.4% for both classifications. In relation to cirrhosiss diagnosis, the mean FI observed was 26.9% for Ishaks score, and 34.5% for METAVIRs classification. There was excellent intraobserver reproducibility. This new method of digital image analysis for liver fibrosis quantification is affordable and was developed using technology that is available worldwide, allowing the precise identification of all the stages of fibrosis in the group of patients studied, with excellent intraobserver reproducibility

Nascimento de bezerros normais após inseminação artificial utilizando espermatozóides criopreservados obtidos de epidídimos refrigerados de bovinos após a morte Birth of normal calves after artificial insemination using cryopreserved spermatozoa obtained from refrigerated epididymides of death bovine

O objetivo deste estudo foi avaliar as características morfológicas e funcionais dos espermatozóides bovinos recuperados de epidídimos resfriados por longos períodos e posteriormente criopreservados. Testículos bovinos foram coletados no abatedouro, transportados ao laboratório e armazenados a 5&deg;C por 0, 24, 48 e 72 horas (n=10 para cada tratamento). Os espermatozóides foram extraídos de cada epidídimo, avaliados e diluídos em meio tris-gema-glicerol a 7% e criopreservados em nitrogênio líquido. As características morfológicas e funcionais dos espermatozóides foram avaliadas in vitro por análise microscópica e in vivo, por meio de inseminação artificial. Foram observadas alterações morfológicas características da imaturidade dos espermatozóides e redução da motilidade após 72 horas de refrigeração dos epidídimos. Esses parâmetros também foram alterados após o descongelamento, em todos os tratamentos. A manutenção dos espermatozoides a 5&deg;C por 72h reduziu a motilidade espermática. Em todos os tratamentos foram observadas alterações morfológicas características da imaturidade dos espermatozoides e redução da motilidade após o descongelamento. A integridade de membrana plasmática e acrossoma somente foram afetadas pós criopreservação nos grupos mantidos a 5&deg;C durante 48 ou 72h antes da criopreservação. Contudo, a capacidade de fecundação dos espermatozóides mantidos a 5&deg;C durante 24 ou 72h antes da criopreservação foi suficiente para promover duas gestações e nascimento de bezerros saudáveis. Esses resultados indicam que a recuperação e a criopreservação de espermatozóides obtidos de epidídimos mantidos a 5&deg;C, até 72h, provenientes de animais mortos é uma opção viável para preservar gametas masculinos para compor um banco de germoplasma.The objective of this study was to evaluate the morphological and functional characteristics of bovine spermatozoa retrieved from chilled epidydimides for long periods and cryopreserved. Bovine testicles were collected in abattoir, transported to the laboratory and stored at 5&deg;C for 0, 24, 48h e 72 hours (n=10 for each storage time treatment group). The spermatozoa were retrieved from each epidydimides, evaluated and diluted in tris-egg yolk-glycerol 7% medium and cryopreserved in liquid nitrogen. The morphological and functional characteristics of the spermatozoa were analyzed in vitro, by microscopic evaluation and in vivo, using artificial insemination. Morphological alterations as sperm immaturity and motility reduction decreased after 72h of epididymides refrigeration and after thaw sperm were observed. The membrane and acrosome integrity were only affected in G48 and G72 groups after cryopreservation. However, the sperm capacity of fertilization post-cryopreservation was sufficient to promote two pregnancies and birth of healthy calves from G24 h and G72h groups. These results indicated that recovery and cryopreservation of chilled epididymal sperm until 72h from dead animals is a viable option to preserve male gametes to compose a germplasm bank

Intravoxel incoherent motion diffusion weighted MR imaging at 3.0 T: assessment of steatohepatitis and fibrosis compared with liver biopsy in type 2 diabetic patients

Pedro Emmanuel Americano Alvarenga do Brasil. Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Documento produzido em parceria ou por autor vinculado à Fiocruz, mas não consta a informação no documento.Submitted by Repositório Arca ([email protected]) on 2019-04-24T17:38:45Z No. of bitstreams: 1 license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5)Approved for entry into archive by Janaína Nascimento ([email protected]) on 2019-08-23T13:37:24Z (GMT) No. of bitstreams: 2 ve_Parente_Daniella_etal_INI_2015.pdf: 1034965 bytes, checksum: 800fda7bd7b6baf2ec28618797467eda (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5)Made available in DSpace on 2019-08-23T13:37:24Z (GMT). No. of bitstreams: 2 ve_Parente_Daniella_etal_INI_2015.pdf: 1034965 bytes, checksum: 800fda7bd7b6baf2ec28618797467eda (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2015D’Or Institute for Research and Education. Rio de Janeiro, RJ, Brazil / Federal University of Rio de Janeiro. Rio de Janeiro, RJ, Brazil.D’Or Institute for Research and Education. Rio de Janeiro, RJ, Brazil / University of São Paulo. Institute of Physics of São Carlos. São Carlos, SP, Brazil.D’Or Institute for Research and Education. Rio de Janeiro, RJ, Brazil.University of the State of Rio de Janeiro. Rio de Janeiro, RJ, Brazil.Federal University of Rio de Janeiro. Rio de Janeiro, RJ, Brazil / University of the State of Rio de Janeiro. Rio de Janeiro, RJ, Brazil.Federal University of São Paulo. São Paulo, SP, Brazil.University of the State of Rio de Janeiro. Rio de Janeiro, RJ, Brazil.D’Or Institute for Research and Education. Rio de Janeiro, RJ, Brazil.University of the State of Rio de Janeiro. Rio de Janeiro, RJ, Brazil.D’Or Institute for Research and Education. Rio de Janeiro, RJ, Brazil / Federal University of Rio de Janeiro. Rio de Janeiro, RJ, Brazil / University of the State of Rio de Janeiro. Rio de Janeiro, RJ, Brazil.D’Or Institute for Research and Education. Rio de Janeiro, RJ, Brazil / Federal University of Rio de Janeiro. Rio de Janeiro, RJ, Brazil

Avaliação genética e fenotípica de características de conformação em potros de três raças eqüinas

Foram ,utilizadas mensurações de 418 potros das raças Quarto de Milha, Mangalarga e Árabe, com o objetivo de estimar herdabilidades e correlações genéticas e fenotípicas entre características de conformação por meio da metodologia da máxima verossimilhança restrita. O arquivo de pedigree era composto de 3.408 animais. Os efeitos das covariáveis grau de endogamia do potro, idade da égua ao parto, duração do período de gestação e idade à mensuração sobre estas estimativas foram avaliados. Consideraram- se também como efeitos fixos rebanho, estação, ano de nascimento, grupo genético e sexo. Nos animais com idade média de 194 dias, foram feitas as seguintes mensurações: altura na cernelha, pedmetro torácico, perímetro da canela, comprimento do corpo e peso corporal. As médias, os erros-padrão e as respectivas herdabilidades (ú ), sem o ajuste de covariável, foram: 125,11 i 0,16 cm e 0,43 para altura da cernelha; 130,34 i 0,30 cm e 0,54 para perímetro torácico; 16,68 .+_ 0,04 cm e 0,38 para perímetro da canela; 121,91 i 0,20 cm e 0,20 para comprimento do corpo e 190,25 i 1,06 kg e 0,61 para peso corporal. Foram observadas pequenas variações nos valores estimados de ú de todas as características, com a inclusão de covariáveis no modelo. Comportamento similar foi observado na análise bicaracterística. As correlações genéticas (Fg ) e fenotípicas (íªp entre as característiças foram, respectivamente, altura da eemelha e perímer torácico (0,58; 0,69), altura ,da cemelha e perímetro da canela (0,87; 0,64), altura da cernclha e comprimento do corpo (0,97; 0,79), altura da cemelha e peso corporal (0,37; 0,63), perimetro torácico e perímetro da canela ( l ,00; 0,66), perímetro torácico e comprimento do corpo (0,61; 0,70), perímetro torácico e peso corporal (1,00; 0,99), perímetro da canela e comprimento do corpo (0,86; 0,60), perímetro da canela e peso corporal (1,00; 0,68) e comprimento do corpo e peso corporal (0,61; 0,67). Pouca variação foi encontrada nas estimativas de Fg e Fp com a inclusão de covariáveis no modelo.Conformation trait measurements of 418 foals, averaging 194 days of age, were used to estimate heritability and genetic and phenotypic correlations, by Restricted Maximum Likelihood methodology. The effects of the covariates inbreeding coefficient, parturition age, gestation length and age of the animal on these estimates were considered in the analyses. Herd, season, year of birth, genetic group and sex were considered fixed effects. The observed mean i standard error and heritability estimates were, respectively, 125.11 i 0.16 cm and 0.43 for wither height; 130.34 i 0.30 cm and 0.54 for toraxic circumference, 16.68 i 0.04 cm and 0.38 for cannon circurnference; 121 .91 i 0.20 cm and 0.20 for body length; and 190.25 i 1.06 kg and 0.61 for body weight. Small changes in the heritability estimates were observed when the covariates were introduced in the A models. Similar results for the heritability estimates of the conformation traits were observed in the single and bitrait analyses. Genetic and phenotypic correlation estimates were obtained for wither height and toraxic circumference (0.58; 0.69), wither height and cannon circumference (0.87; 0.64), wither height and body length (0.97; 0.79), wither height and body weight (0.37; 0.63), toraxic circumference and cannon circumference (1,00; 0.66), toraxic circnmference and body length (0.61; 0.70), toraxic circumference and body weight (1 .00; 0.99), cannon circumference and boby length (0.86;0.66), cannon circuference and body weigth (1.00;0.68), and body length and body weigth (0.61; 0.67). Small variations in the correlation coefficient were observed when the covariantes were included in the models

Pure molecular diffusion (D), perfusion-related diffusion (D*), and perfusion fraction (f) in histological fibrosis stage groups (F0 to F4).

<p>D, pure molecular diffusion; D*, perfusion-related diffusion; f, vascular fraction; SD; Standard deviation; IQR; interquartil range; NA, not assigned</p><p>Pure molecular diffusion (D), perfusion-related diffusion (D*), and perfusion fraction (f) in histological fibrosis stage groups (F0 to F4).</p