The last decade has seen a sharp increase in the number of scientific publications describing physiological and pathological functions of extracellular vesicles (EVs), a collective term covering various subtypes of cell-released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, and many other names. However, specific issues arise when working with these entities, whose size and amount often make them difficult to obtain as relatively pure preparations, and to characterize properly. The International Society for Extracellular Vesicles (ISEV) proposed Minimal Information for Studies of Extracellular Vesicles (“MISEV”) guidelines for the field in 2014. We now update these “MISEV2014” guidelines based on evolution of the collective knowledge in the last four years. An important point to consider is that ascribing a specific function to EVs in general, or to subtypes of EVs, requires reporting of specific information beyond mere description of function in a crude, potentially contaminated, and heterogeneous preparation. For example, claims that exosomes are endowed with exquisite and specific activities remain difficult to support experimentally, given our still limited knowledge of their specific molecular machineries of biogenesis and release, as compared with other biophysically similar EVs. The MISEV2018 guidelines include tables and outlines of suggested protocols and steps to follow to document specific EV-associated functional activities. Finally, a checklist is provided with summaries of key points.
Retroviruses have been invading mammalian germlines for millions of years, accumulating in the form of endogenous retroviruses (ERVs) that account for nearly one-tenth of the mouse and human genomes. ERVs are epigenetically silenced during development, yet the cellular factors recognizing ERVs in a sequence-specific manner remain elusive. Here we demonstrate that ZFP809, a member of the Kr€ uppel-associated box zinc finger protein (KRAB-ZFP) family, initiates the silencing of ERVs in a sequence-specific manner via recruitment of heterochromatin-inducing complexes. ZFP809 knockout mice display highly elevated levels of ZFP809-targeted ERVs in somatic tissues. ERV reactivation is accompanied by an epigenetic shift from repressive to active histone modifications but only slight destabilization of DNA methylation. Importantly, using conditional alleles and rescue experiments, we demonstrate that ZFP809 is required to initiate ERV silencing during embryonic development but becomes largely dispensable in somatic tissues. Finally, we show that the DNA-binding specificity of ZFP809 is evolutionarily conserved in the Muroidea superfamily of rodents and predates the endogenization of retroviruses presently targeted by ZFP809 in Mus musculus. In sum, these data provide compelling evidence that ZFP809 evolved to recognize foreign DNA and establish histone modification-based epigenetic silencing of ERVs.
-Data from 359 Holstein and 54 Jersey cows (total of 1359 observations) collected in commercial herds in the northeast of Brazil were used to evaluate six environmental stress indexes. The selection criterion was the correlation between the index value with the rectal temperature (RT) and the respiratory rate (RR) of cows. Both Temperature-Humidity Index (THI) and Black Globe-Humidity Index (BGHI) had the lowest correlations with animal RT and RR. The selected indexes were the Equivalent Temperature Index (r = 0.293 and 0.520 with RT and RR, respectively) and the Heat Load Index (0.286 and 0.542, respectively).
RESUMO -As características do pelame (espessura da capa, comprimento médio dos pêlos, número de pêlos por unidade de área, densidade de massa dos pêlos, ângulo de inclinação dos pêlos com respeito a superfície da epiderme e diâmetro médio do pêlos) foram avaliadas em 973 vacas da raça Holandesa, entre novembro de 2000 e abril de 2001, numa área localizada 20cm abaixo da coluna vertebral, no centro do tronco, tanto nas malhas brancas como nas negras. As amostras de pêlos foram obtidas com um alicate comum adaptado. O método da Máxima Verossimilhança Restrita (REML) foi usado para estimar os componentes de variância e covariância sob modelo animal, sendo empregado o sistema MTDFREML. Os resultados mostraram que as características do pelame preto são diferentes das do branco, quando os animais são criados em ambiente tropical. O pelame preto apresentou-se menos denso, com pêlos mais curtos e grossos devido à maior necessidade de perder calor, enquanto que, o pelame branco é mais denso e com pêlos mais compridos, oferecendo uma melhor proteção contra à radiação solar direta. A seleção de vacas predominantemente negras pode ser uma boa escolha para aumentar a resistência do gado Holandês às condições do ambiente tropical, principalmente à radiação solar, quando esses animais são criados a campo, devido a que a epiderme sob esse tipo de malhas é altamente pigmentada. Tal seleção pode ser facilmente realizada, considerando a alta herdabilidade (h 2 =0,75) para a proporção de malhas negras. Esta seleção deve ser realizada no sentido de um pelame menos denso, com pêlos curtos e grossos favorecendo as perdas de calor sensível e calor latente.Palavras-chaves: adaptação, bovinos, raça Holandesa, pelame, seleção Haircoat Traits in Holstein Cows in Tropical Environments: a Genetic and Adaptative StudyABSTRACT -The traits of the haircoat (haircoat thickness, average hair lenght with respect skin surface, number of hair per unit area, hair mass density, hair angle and average hair diameter) were estimated in 973 Holstein cattle in the period between November 2000 and April of 2001. These traits were measured in an area 20cm below the dorsal line in the center of the thorax, for the black as for the white haircoat. Hair samplings were performed with adapted pliers. The method of Restricted Maximum Likelihood (REML) was used to estimate the variance and covariance components under the Animal model, the program used being the MTDFREML. The results showed that traits of black haircoat were different from that of the white haircoat, when the animals are breed in a tropical environment. Black haircoat was less dense, with shorter and ticker hair fibres in order to ease the heat dissipation, while the white haircoat was denser with longer hairs, which favored protection against direct solar radiation. The selection of predominantly black cows would be a good choice under the strong solar radiation in tropical environment, because the pigmented skin under this type coat. Such a selection could be easily performed considering the high h...
BackgroundVarious studies showed that chemotherapy can control schistosomiasis morbidity, but association of measures (water supply, sewage disposal and increase of socioeconomic conditions) is necessary for transmission control.Methodology/Principal FindingsA survey dealing with socioeconomic conditions, snail survey, contact with natural waters, and clinical and stool examinations was undertaken at an endemic area in the State of Minas Gerais, Brazil. The methodology used was the same for both evaluations (1981 and 2005). Four hundred and seventy-five out of 1,474 individuals studied in 1981 could be contacted. From these, 358 were submitted to stool examination, and 231 of them were clinically examined. Patients eliminating S. mansoni eggs in their stools were treated. The results showed that the prevalence rate in Comercinho, a municipality of the State of Minas Gerais, Brazil, was substantially reduced to 70.4% and 1.7% in 1981 and 2005, respectively, as well as the frequency of the hepatosplenic form (7% to 1.3%) after five treatments effectuated between 1981 and 1992. No other new case of this form was detected from 1981 onwards. Another important aspect to be considered was the improvement of people's living standard that occurred in the region after more than two decades' efforts (better housing, professional skill and adequate basic sanitation).Conclusion/SignificanceThe control of morbidity and very significant decrease of schistosomiasis transmission in an area until then considered as hyperendemic was possible by means of association of successive specific treatments of the local population, together with the construction of privies, water supply in the houses and improvement of socioeconomic conditions.
RESUMO -É apresentado um método para a estimativa do balanço térmico por radiação em vacas da raça Holandesa expostas ao sol e sob a sombra em uma pastagem, com o objetivo de estabelecer um padrão morfológico mais adequado para esses animais em região tropical. O método envolve a determinação das características do pelame (área de superfície de malhas negras, espessura da capa, comprimento e diâmetro dos pêlos, número de pêlos por unidade de área), temperatura da superfície da capa e da epiderme e variáveis ambientais (irradiância solar, temperatura do ar e de globo, vento). Foi determinada a quantidade de radiação de ondas curtas efetivamente transmitida através do pelame e que atinge as camadas profundas da epiderme. Demonstrou-se, por intermédio de um exemplo prático, que as áreas de pelame negro apresentaram balanço radiante mais elevado que as áreas brancas, mas estas últimas possuiam maior transmissividade para a radiação de ondas curtas. Animais predominantemente negros foram melhor protegidos contra a radiação de ondas curtas que os predominantemente brancos, especialmente quando a capa de pelame é pouco espessa (até 4 a 7 mm), mais adequada para ambientes tropicais.Palavras-chave: balanço térmico, pelame, radiação, vacas Holandesas Estimate of Radiation Heat Balance of Holstein Cows in the Sun and under the Shade in a Tropical EnvironmentABSTRACT -A method is presented to estimate the radiation heat balance of Holstein cows exposed to sun and under shade of a pasture, to contribute with a morphological characteristics better adequate for these animals in a tropical environment. The methods involved the haircoat characteristics (surface area of black spots, coat thickness, hair length, hair diameter, number of hairs per unit of area), skin and coat surface temperatures and environmental variables (solar irradiance, wind speed, air and globe temperatures). The amount of shortwave energy which effectively is transmitted through the haircoat and that reach deply the epiderms layers was obtained. It was demonstrated by a practical example that the black areas of the coat presented higher radiation heat balance than that of the white areas, but those had greater transmissivity for shortwave radiation. Predominantly black cows were better protected against shortwave radiation than the white ones, especially if the animals presented thin (4 to 7 mm or less) haircoats, the most suitable for tropical environments.Key Words: haircoat, Holstein cows, radiation balance Rev. bras. zootec., v.28, n.6, p.1403-1411, 1999 1 DSc, Livre-Docente, Professor Titular de Bioclimatologia, Departamento de Zootecnia, Faculdade de Ciências Agrárias e Veterinárias, UNESP, 14870-000 Jaboticabal, SP. E-mail: rgomes@fcav.unesp.br IntroduçãoA termorregulação nos animais tem sido extensivamente estudada em câmaras climáticas e calorímetros, nos quais as condições ambientes podem ser facilmente controladas. Entretanto, o ambiente externo, no qual os animais vivem normalmente, é muito mais complexo, pois fatores como velocidade do vento, r...
RNA-based therapies have great potential to treat many undruggable human diseases. However, their efficacy, in particular for mRNA, remains hampered by poor cellular delivery and limited endosomal escape. Development and optimisation of delivery vectors, such as lipid nanoparticles (LNPs), are impeded by limited screening methods to probe the intracellular processing of LNPs in sufficient detail. We have developed a high-throughput imaging-based endosomal escape assay utilising a Galectin-9 reporter and fluorescently labelled mRNA to probe correlations between nanoparticle-mediated uptake, endosomal escape frequency, and mRNA translation. Furthermore, this assay has been integrated within a screening platform for optimisation of lipid nanoparticle formulations. We show that Galectin-9 recruitment is a robust, quantitative reporter of endosomal escape events induced by different mRNA delivery nanoparticles and small molecules. We identify nanoparticles with superior escape properties and demonstrate cell line variances in endosomal escape response, highlighting the need for fine-tuning of delivery formulations for specific applications.
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