A CADEIA DE CUSTÓDIA NA COLETA DA PROVA DIGITAL DE ACORDO COM A LEI 13.964/2019, DOS SEUS ARTIGOS 158-A AO 158-F

The present study aimed to identify the procedures necessary to carry out the collection of digital evidence and report the risk factors that can harm the digital evidence obtained on the understanding of scholars about the valuation or nullity of evidence that had no chain of custody or there was violation of your chain of custody. For this, bibliographic research was used. It was found that due to the nature of digital evidence, it was necessary to standardize its treatment in order to guarantee its integrity and authenticity. Regarding the positions of the eminent authors regarding the solution of the breach of the chain of custody, they are frontally contrary, inclining the majority doctrine in the sense that any vices in the chain of custody do not, in and of themselves, entail the inadmissibility or nullity of the test.El presente estudio tuvo como objetivo identificar los procedimientos necesarios para realizar la recolección de pruebas digitales y reportar los factores de riesgo que pueden perjudicar la prueba digital obtenida al entender a los adoctrinadores sobre la valoración o nulidad de las pruebas que no tenían cadena de custodia o tenían una violación de su cadena de custodia. Para ello, se utilizó la revisión bibliográfica y la investigación documental con un enfoque inductivo. Se determinó que, debido a la naturaleza de las pruebas digitales, era necesario normalizar su tratamiento a fin de garantizar su integridad y autenticidad. Sobre las posiciones de los eminentes autores en cuanto a la solución de la ruptura de la cadena de custodia, son frontalmente contrarias, inclinando la doctrina mayoritaria en el sentido de que los eventuales vicios en la cadena de custodia no implican, en plan y por sí mismo, la inadmisibilidad o nulidad de las pruebas.O presente estudo teve objetivo de identificar os procedimentos necessários para a realização da coleta da prova digital e relatar os fatores de risco que podem prejudicar a prova digital obtida sobre entendimento dos doutrinadores sobre a valoração ou nulidade da prova que não teve cadeia de custódia ou houve violação da sua cadeia de custódia. Para tal foram utilizadas pesquisas bibliográficas. Constatou-se que em virtude da natureza da evidência digital, foi necessário padronizar o seu tratamento a fim de garantir sua integridade e autenticidade. Sobre as posições dos eminentes autores quanto à solução da quebra da cadeia de custódia, são frontalmente contrárias, inclinando a doutrina majoritária no sentido de que eventuais vícios na cadeia de custódia não acarretam, de plano e por si só, a inadmissibilidade ou a nulidade da prova

In vitro culture of ovine mammary gland cells expressing beta-lactoglobulin and beta-casein

The expression of milk proteins in vitro is essential to exploit the mammary gland cells as a biological model. Enzymatic tissue disaggregation has been widely used to establish mammary cell culture, but its effect in long-term ovine mammary cell culture is not completely elucidated. This study aimed at comparing mechanical/enzymatic and mechanical dissociation methods to establish ovine mammary cell culture. We compared cellular differentiation induced by lactating ewe sérum or fetal bovine serum based on the gene expression levels of milk proteins (beta-lactoglobulin, alpha s1-casein, and betacasein). Mechanically dissociated cells were positive immunostaining for cytokeratin 8.13, such as mammary epithelial cells. These cells are responsible for milk protein expression and they are low immunostaining for vimentin, mesenchymal marker. Mechanical/enzymatic dissociation cells were positive for vimentin. The fastest cell growth (cell/hour) was observed in the mechanical dissociation group cultured with 10% fetal bovine serum medium. Mechanically and mechanically/enzymatically derived cells were able to express beta-casein and beta-lactoglobulin, but not alpha s1-casein. The relative expression of beta-lactoglobulin was not affected by the tissue dissociation method or culture media, beta-casein relative expression was down regulated in mechanically dissociated cells cultured in the presence of lactating ewe serum, (P = 0.019). Beta-casein relative expression was also down regulated in mechanically/enzymatically dissociated cells cultured with fetal bovine sérum (P = 0.021). In the present conditions, we conclude that mechanical dissociation followed by culture with 10% of fetal bovine serum was the most efficient method to induce milk proteins’ mRNA expression by ovine mammary epithelial cells in vitro.A expressão in vitro de proteínas do leite e essencial para explorar as células da glândula mamaria como um modelo biológico. A desagregação tecidual via enzimática e amplamente utilizada para o estabelecimento cultivo de células mamarias. No entanto, seu efeito a longo prazo no cultivo de células da glândula mamaria ovina ainda não e bem elucidado. Este estudo tem como objetivo comparar dois métodos de dissociação tecidual, mecânico/enzimático e mecânico, para estabelecer cultivo celular de glândula mamaria ovina. A indução da diferenciação celular, por adição de soro de ovelha lactante ou soro fetal bovino, foi avaliada pelos níveis de expressão de proteínas do leite (beta-lactoglobulina, alpha s1-caseina e beta-caseína). Células mecanicamente dissociadas foram positivamente marcadas para a presença de citoqueratina 8.13, marcador para células epiteliais mamarias. Essas células são as responsáveis pela produção das proteínas do leite e são pouco marcadas para a presença de vimentina, marcador para células de origem mesenquimal. Já as células obtidas da dissociação mecânica/ enzimática foram positivamente marcadas para presença de vimentina. A maior velocidade de crescimento (células/hora) foi observado para o grupo com dissociação mecânica cultivado em meio com 10% de soro fetal bovino. As células obtidas tanto da dissociação mecânica quanto mecânica/enzimática foram capazes de expressar beta-lactoglobulina e beta-caseína, mas não alfa s1-caseina. A expressão relativa de beta-lactoglobulina não foi afetada pelo método de dissociação ou meio de cultivo. A expressão relativa da beta-caseína foi negativamente regulada para células mecanicamente dissociadas e cultivadas na presença de soro de ovelha lactante (P = 0,019). A expressão relativa da beta-caseína também foi negativamente regulada para células dissociadas de forma mecânica/enzimática e cultivadas com soro fetal bovino (P = 0,021). Nas condições do presente estudo, concluímos que o método de dissociação mecânica seguido pelo cultivo em meio com 10% de soro fetal bovino foi o método mais eficiente para induzir a expressão mRNA de proteínas do leite por células epiteliais mamarias ovinas in vitro

AVALIAÇÃO DISTRIBUIÇÃO MITOCONDRIAL NA MATURAÇÃO IN VITRO DE OÓCITOS BOVINOS DE OVÁRIOS COM BAIXA E ALTA CONTAGEM DE FOLÍCULOS ANTRAIS.

In vitro embryo production (PIVE) depends on the stages of maturation, fertilization and in vitro cultivation, being the decisive stage for the oocyte to reach the morula and blastocyst stage. This study aimed to evaluate the quality of in vitro maturation, according to the mitochondrial distribution of oocytes from ovaries of slaughter females with low or high antral follicle count (CFA). A total of 243 ovaries were aspirated from which 334 oocytes were selected for five replicates for each group. The ovaries were separated into two groups: 1- low CFA (n = 19) 2- high CFA (n = 53) and stained with the Mito Traker Red probe. Data obtained were analyzed by Fisher's exact nonparametric test ( α = 5%) using the Statistical Analysis System for Windows software (SAS Inst., Inc., Cary, NC). Regarding the location of mitochondria, the present study showed significant differences (p <0.001) between immature and mature oocytes, which is desirable. Mature oocytes had a higher percentage of dispersed mitochondria in the cytoplasm and immature oocytes in the periphery, regardless of the experimental group (low and high CFA). In immature oocytes, there was no difference (p = 0.1542) in the location of mitochondria between the low and high CFA groups. However, the oocytes of the low CFA group, after maturation, presented smaller number of oocytes with mitochondria in the periphery compared to the high CFA (p = 0.0355), indicating for this parameter a better competence of these oocytes for PIVE.A Produção in vitro de embriões (PIVE) depende das etapas de maturação, fertilização e cultivo in vitro, sendo a maturação a etapa decisiva para o oócito atingir o estádio de mórula e blastocisto. Neste trabalho objetivou-se avaliar a qualidade de maturação in vitro, de acordo com a distribuição mitocondrial de oócitos provenientes de ovários de fêmeas de abatedouro, com baixa ou alta contagem de folículos antrais (CFA). Foram aspirados 243 ovários dos quais selecionou-se 334 oócitos perante de cinco repetições para cada grupo. Os ovários foram separados em dois grupos: 1- baixa CFA (n=19), 2- alta CFA (n= 53) e os oócitos recuperados corados com a sonda MitoTracker Red. Os dados obtidos foram analisados pelo teste não paramétrico exato de Fisher (α=5%) mediante o software Statistical Analysis System for Windows (SAS Inst., Inc., Cary, NC). Em relação a localização das mitocôndrias o presente trabalho apresentou diferenças (p<0,001) entre oócitos imaturos e maturados, o que é desejável. Oócitos maturados apresentaram maior porcentagem de mitocôndrias dispersas no citoplasma e oócitos imaturos na periferia, independente do grupo experimental (baixa e alta CFA). Em oócitos imaturos, não houve diferença (p=0,1542) na localização das mitocôndrias entre os grupos baixa e alta CFA. No entanto, o grupo baixa CFA, após a maturação, apresentou menor quantidade de oócitos com mitocôndrias na periferia em relação aos de alta CFA (p = 0,0355), indicando melhor aquisição de competência desses oócitos para PIVE

Otoliths-composed gelatin/sodium alginate scaffolds for bone regeneration

Evidence that otoliths, mineral-rich limestone concrescences present in the inner ear of bone fishes, can accelerate bone formation in vivo has been previously reported. The goal of this work was the development, characterization, and evaluation of the cytocompatibility of otoliths-incorporated sodium alginate and gelatin scaffolds. Cynoscion acoupaderived otoliths were characterized by X-ray fluorescence spectrometry (FRX), particle size, free lime, and weight loss by calcination. Furthermore, otoliths were incorporated into sodium alginate (ALG/OTL-s) or gelatin (GEL/OTL-s) scaffolds, previously developed by freeze-drying. Then, the scaffolds were characterized by thermogravimetric analysis (TGA/DTG), differential scanning calorimetry (DSC), infrared spectroscopy with Fourier transform (FTIR), swelling tests, and scanning electron microscopy (SEM). Cytotoxicity assays were run against J774.G8 macrophages and MC3T3-E1 osteoblasts. Data obtained from TGA/DTG, DSC, and FTIR analyses confirmed the interaction between otoliths and the polymeric scaffolds. SEM showed the homogeneous porous 3D structure rich in otolith micro-fragments in both scaffolds. Swelling of the GEL/OTL-s (63.54±3.0%) was greater than of ALG/OTL-s (13.36±9.9%) (p0.05) and significantly higher than that treated with Triton-X (p0.05). However, by 48 h, only ALG/OTL-s showed growth similar to control (p>0.05), whereas GEL/OTL showed a significantly lower growth index (p<0.05). In conclusion, the physicochemical profiles suggest proper interaction between the otoliths and the two developed polymeric 3D scaffolds. Moreover, both materials showed cytocompatibility with J774.G8 macrophages but the growth of MC3T3-E1 osteoblasts was higher when exposed to ALG/OTL-s. These data suggest that sodium alginate/otoliths scaffolds are potential biomaterials to be used in bone regeneration applications.We would like to thank the National Council for Scientific and Technological Development (CNPq) and the Foundation for Research and Technological Innovation Support of the State of Sergipe for the financial support in this study. EMBS acknowledges the sponsorship of the projects M-ERA-NET-0004/2015-PAIRED and UIDB/04469/2020 (strategic fund), received support from the Portuguese Science and Technology Foundation, Ministry of Science and Education (FCT/MEC) through national funds, and was co-financed by FEDER, under the Partnership Agreement PT2020.info:eu-repo/semantics/publishedVersio

Photography-based taxonomy is inadequate, unnecessary, and potentially harmful for biological sciences

The question whether taxonomic descriptions naming new animal species without type specimen(s) deposited in collections should be accepted for publication by scientific journals and allowed by the Code has already been discussed in Zootaxa (Dubois & Nemésio 2007; Donegan 2008, 2009; Nemésio 2009a–b; Dubois 2009; Gentile & Snell 2009; Minelli 2009; Cianferoni & Bartolozzi 2016; Amorim et al. 2016). This question was again raised in a letter supported by 35 signatories published in the journal Nature (Pape et al. 2016) on 15 September 2016. On 25 September 2016, the following rebuttal (strictly limited to 300 words as per the editorial rules of Nature) was submitted to Nature, which on 18 October 2016 refused to publish it. As we think this problem is a very important one for zoological taxonomy, this text is published here exactly as submitted to Nature, followed by the list of the 493 taxonomists and collection-based researchers who signed it in the short time span from 20 September to 6 October 2016

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

Measurement of the top quark forward-backward production asymmetry and the anomalous chromoelectric and chromomagnetic moments in pp collisions at √s = 13 TeV

Abstract The parton-level top quark (t) forward-backward asymmetry and the anomalous chromoelectric (d̂ t) and chromomagnetic (μ̂ t) moments have been measured using LHC pp collisions at a center-of-mass energy of 13 TeV, collected in the CMS detector in a data sample corresponding to an integrated luminosity of 35.9 fb−1. The linearized variable AFB(1) is used to approximate the asymmetry. Candidate t t ¯ events decaying to a muon or electron and jets in final states with low and high Lorentz boosts are selected and reconstructed using a fit of the kinematic distributions of the decay products to those expected for t t ¯ final states. The values found for the parameters are AFB(1)=0.048−0.087+0.095(stat)−0.029+0.020(syst),μ̂t=−0.024−0.009+0.013(stat)−0.011+0.016(syst), and a limit is placed on the magnitude of | d̂ t| &lt; 0.03 at 95% confidence level. [Figure not available: see fulltext.