Estudo do desenvolvimento extra-uterino de Didelphis aurita Wied, 1826, em cativeiro: investigação de critérios para estimativa de idade

The growth and development of 37 litters of Didelphis aurita were studied in order to establish a method for age estimation. The sample included 11 litters of captive females and 26 litters captured as pouch young in the State of Rio de Janeiro, Brazil, which were subsequently reared in captivity. Serial measurements of body (snout-rump) and tail lenghts and weight, together with sequential observations of development of selected external characters, were made from the first day to one year of age. The life cycle was divided into three periods: pouch life (1-100 days); weaning (100-150 days) and puberty period (150 days on). Nine dental classes were assigned to all spec1mens based on the time of molar erupt1on and replacement of third -premolar. No clear sex differences were observed regarding dental eruption sequences, but in males of Class VI body and tail lengths and weights were significantly greater than in females. Dental eruptlon sequence did not differ between captive and wild caught animals,suggesing tnat env1ronmental conditions had little or no effect on dental development. The study of weight var1ation using individuals of known age, demonstrated that its is not a good age indicator because of high variance. An empirical multiple regression equation was calculated for age estimation using the selected variables. The standard error using random samples was within eight days. Only animals older than 18 days can nave their age assessed by this equation.O crescimento e desenvolvimento de 37 ninhadas de Dídelphis aurita foram estudadas objetivando estabelecer um método para estimativa de idade. O grupo em estudo constitui-se de 11 ninhadas nascidas de fêmeas criadas em cativeiro e 26 ninhadas capturadas no marsupio de fêmeas provindas do Estado do Rio de Janeiro, Brasil, e mantidas em cativeiro. Periodicamente foram registradas medidas de corpo, cauda e peso junto com observações sequenciais de caracteres externos selecionados, durante um ano de vida, em animais vivos. O ciclo de vida desses animais foi dividido em três períodos: a vida no marsúpio (1-100 dias), desmamados (100-150 dias) e puberdade (depois de 150 dias). Para as duas amostras foram estabelecidas nove classes dentarias baseadas no tempo de erupção dos molares e troca do terceiro premolar. Não foram evidenciadas nítidas diferenças no crescimento de machos e fêmeas nas diferentes classes dentarias, porém os machos da classe VI têm suas medidas significativamente maiores do que as fêmeas. A sequência de erupção dentaria foi a ·mesma observada entre os animais de campo e cativeiro, sugerindo. que as condições ambientais tiveram um pequeno ou nenhum efeito sobre o desenvolvimento dentário. Entre os espécimes de idade conhecida, o estudo do peso demonstrou que é o parâmetro de maior variação individual, não sendo um bom índice para estimativa de idade. Baseando-se em variáveis selec1onadas, calculou-se uma equação de regressão múltipla escalonada para estimativa idade. Usando esta equação em uma amostra randômica, o erro padrão verificado foi de 8 dias. A aplicação desta equação só é possível a partir dos 18 dias de idade

Indústria 4.0 e economia circular: uma transformação digital e sustentável na engenharia, com aplicação no setor de alimentos e bebidas

O desenvolvimento econômico e o crescimento populacional acarretaram um aumento contínuo na demanda por bens e serviços, que propiciam o consumo insustentável dos recursos naturais. Dessa forma, novas estratégias para o uso eficiente desses recursos surgem a cada dia, a fim de manter uma economia sustentável a longo prazo. Neste contexto, o presente estudo apresenta uma análise de duas correntes que permeiam o setor industrial e ganham destaque na resolução desses desafios, são elas: a Iindústria 4.0 e a Economia Circular. A pesquisa realizada a baseia em uma revisão bibliográfica sobre os temas, e na confecção de um estudo de caso aplicado a uma indústria do setor de alimentos e bebidas, produtora de suco de laranja integral. Com o intuito de compreender se e como as práticas de digitalização e circularidade são coerentes quando aplicadas em conjunto, são apresentadas possíveis interferências no processo produtivo da Citrino (empresa produtora de suco de laranja), que envolvem a aplicação dessas duas correntes. Ao final, analisa-se qualitativamente se os impactos dessas interferências são harmônicos, e se elas levam o modelo de negócio da empresa a um patamar mais sustentável. No caso apresentado, as interferências não só podem ser aplicadas em conjunto, como é provável que elas se potencializem. No entanto, a convergência das correntes não é garantida, pois é fundamental que haja uma política corporativa forte no intuito de tornar a digitalização da indústria mais sustentável

Derivation Of Functional Human Astrocytes From Cerebral Organoids

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Astrocytes play a critical role in the development and homeostasis of the central nervous system (CNS). Astrocyte dysfunction results in several neurological and degenerative diseases. However, a major challenge to our understanding of astrocyte physiology and pathology is the restriction of studies to animal models, human post-mortem brain tissues, or samples obtained from invasive surgical procedures. Here, we report a protocol to generate human functional astrocytes from cerebral organoids derived from human pluripotent stem cells. The cellular isolation of cerebral organoids yielded cells that were morphologically and functionally like astrocytes. Immunolabelling and proteomic assays revealed that human organoid-derived astrocytes express the main astrocytic molecular markers, including glutamate transporters, specific enzymes and cytoskeletal proteins. We found that organoid-derived astrocytes strongly supported neuronal survival and neurite outgrowth and responded to ATP through transient calcium wave elevations, which are hallmarks of astrocyte physiology. Additionally, these astrocytes presented similar functional pathways to those isolated from adult human cortex by surgical procedures. This is the first study to provide proteomic and functional analyses of astrocytes isolated from human cerebral organoids. The isolation of these astrocytes holds great potential for the investigation of developmental and evolutionary features of the human brain and provides a useful approach to drug screening and neurodegenerative disease modelling.7Fundacao Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de JaneiroFAPERJConselho Nacional para o Desenvolvimento Cientifico e TecnologicoCNPqCoordenacao de Aperfeicoamento de Pessoal de Nivel SuperiorCAPESFundacao de Amparo a Pesquisa do Estado de Sao PauloFAPESP [13/08711-3, 14/10068-4, 14/21035-0, 16/07332-7]Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Thyroid hormone treated astrocytes induce maturation of cerebral cortical neurons through modulation of proteoglycan levels

Proper brain neuronal circuitry formation and synapse development is dependent on specific cues, either genetic or epigenetic, provided by the surrounding neural environment. Within the sesignals, thyroid hormones (T3 and T4) play crucial role in several steps of brain morphogenesis including proliferation of progenitor cells, neuronal differentiation, maturation, migration, and synapse formation. the lack of thyroid hormones during childhood is associated with several impair neuronal connections, cognitive deficits, and mental disorders. Many of the thyroid hormones effects are mediated by astrocytes, although the mechanisms underlying these events are still unknown. in this work, we investigated the effect of 3,5,3'-triiodothyronine-treated (T3-treated) astrocytes on cerebral cortex neuronal differentiation. Culture of neural progenitors from embryonic cerebral cortex mice onto T3-treated astrocyte monolayers yielded an increment in neuronal population, followed by enhancement of neuronal maturation, arborization and neurite outgrowth. in addition, real time PCR assays revealed an increase in the levels of the heparan sulfate proteoglycans, Glypican 1(GPC-1) and Syndecans 3 e 4 (SDC-3 e SDC-4), followed by a decrease in the levels of the chondroitin sulfate proteoglycan, Versican. Disruption of glycosaminoglycan chains by chondroitinase AC or heparanase III completely abolished the effects of T3-treated astrocytes on neuronal morphogenesis. Our work provides evidence that astrocytes are key mediators of T3 actions on cerebral cortex neuronal development and identified potential molecules and pathways involved in neurite extension; which might eventually contribute to a better understanding of axonal regeneration, synapse formation, and neuronal circuitry recover.Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ)Conselho Nacional para o Desenvolvimento Cientifico e TecnologicoCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Univ Fed Rio de Janeiro, Inst Ciencias Biomed, BR-21949590 Rio de Janeiro, RJ, BrazilUniv Fed Rio de Janeiro, Inst Bioquim Med, BR-21949590 Rio de Janeiro, RJ, BrazilUniv Fed Rio de Janeiro, Hosp Univ Clementino Fraga Filho, BR-21949590 Rio de Janeiro, RJ, BrazilUniversidade Federal de São Paulo, Dept Bioquim, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Bioquim, São Paulo, BrazilWeb of Scienc

Transcriptomics reveal an integrative role for maternal thyroid hormones during zebrafish embryogenesis

Thyroid hormones (THs) are essential for embryonic brain development but the genetic mechanisms involved in the action of maternal THs (MTHs) are still largely unknown. As the basis for understanding the underlying genetic mechanisms of MTHs regulation we used an established zebrafish monocarboxylic acid transporter 8 (MCT8) knock-down model and characterised the transcriptome in 25hpf zebrafish embryos. Subsequent mapping of differentially expressed genes using Reactome pathway analysis together with in situ expression analysis and immunohistochemistry revealed the genetic networks and cells under MTHs regulation during zebrafish embryogenesis. We found 4,343 differentially expressed genes and the Reactome pathway analysis revealed that TH is involved in 1681 of these pathways. MTHs regulated the expression of core developmental pathways, such as NOTCH and WNT in a cell specific context. The cellular distribution of neural MTH-target genes demonstrated their cell specific action on neural stem cells and differentiated neuron classes. Taken together our data show that MTHs have a role in zebrafish neurogenesis and suggest they may be involved in cross talk between key pathways in neural development. Given that the observed MCT8 zebrafish knockdown phenotype resembles the symptoms in human patients with Allan-Herndon-Dudley syndrome our data open a window into understanding the genetics of this human congenital condition.Portuguese Fundacao para Ciencia e Tecnologia (FCT) [PTDC/EXPL/MARBIO/0430/2013]; CCMAR FCT Plurianual financing [UID/Multi/04326/2013]; FCT [SFRH/BD/111226/2015, SFRH/BD/108842/2015, SFRH/BPD/89889/2012]; FCT-IF Starting Grant [IF/01274/2014]info:eu-repo/semantics/publishedVersio

Organs to Cells and Cells to Organoids: The Evolution of in vitro Central Nervous System Modelling

With 100 billion neurons and 100 trillion synapses, the human brain is not just the most complex organ in the human body, but has also been described as “the most complex thing in the universe.” The limited availability of human living brain tissue for the study of neurogenesis, neural processes and neurological disorders has resulted in more than a century-long strive from researchers worldwide to model the central nervous system (CNS) and dissect both its striking physiology and enigmatic pathophysiology. The invaluable knowledge gained with the use of animal models and post mortem human tissue remains limited to cross-species similarities and structural features, respectively. The advent of human induced pluripotent stem cell (hiPSC) and 3-D organoid technologies has revolutionised the approach to the study of human brain and CNS in vitro, presenting great potential for disease modelling and translational adoption in drug screening and regenerative medicine, also contributing beneficially to clinical research. We have surveyed more than 100 years of research in CNS modelling and provide in this review an historical excursus of its evolution, from early neural tissue explants and organotypic cultures, to 2-D patient-derived cell monolayers, to the latest development of 3-D cerebral organoids. We have generated a comprehensive summary of CNS modelling techniques and approaches, protocol refinements throughout the course of decades and developments in the study of specific neuropathologies. Current limitations and caveats such as clonal variation, developmental stage, validation of pluripotency and chromosomal stability, functional assessment, reproducibility, accuracy and scalability of these models are also discussed