Subcutaneous adipose tissue classification

The developments in the technologies based on the use of autologous adipose tissue attracted attention to minor depots as possible sampling areas. Some of those depots have never been studied in detail. The present study was performed on subcutaneous adipose depots sampled in different areas with the aim of explaining their morphology, particularly as far as regards stem niches. The results demonstrated that three different types of white adipose tissue (WAT) can be differentiated on the basis of structural and ultrastructural features: deposit WAT (dWAT), structural WAT (sWAT) and fibrous WAT (fWAT). dWAT can be found essentially in large fatty depots in the abdominal area (periumbilical). In the dWAT, cells are tightly packed and linked by a weak net of isolated collagen fibers. Collagenic components are very poor, cells are large and few blood vessels are present. The deep portion appears more fibrous then the superficial one. The microcirculation is formed by thin walled capillaries with rare stem niches. Reinforcement pericyte elements are rarely evident. The sWAT is more stromal; it is located in some areas in the limbs and in the hips. The stroma is fairly well represented, with a good vascularity and adequate staminality. Cells are wrapped by a basket of collagen fibers. The fatty depots of the knees and of the trochanteric areas have quite loose meshes. The fWAT has a noteworthy fibrous component and can be found in areas where a severe mechanic stress occurs. Adipocytes have an individual thick fibrous shell. In conclusion, the present study demonstrates evident differences among subcutaneous WAT deposits, thus suggesting that in regenerative procedures based on autologous adipose tissues the sampling area should not be randomly chosen, but it should be oriented by evidence based evaluations. The structural peculiarities of the sWAT, and particularly of its microcirculation, suggest that it could represent a privileged source for regenerative procedures based on autologous adipose tissues

The trochanteric fat pad

Technological developments based on the use of autologous white adipose tissue (WAT) attracted attention to minor fat depots as possible sources of adipose tissue. In plastic surgery, the trochanteric fatty pad is one of the most used WAT depots for its location and organoleptic characteristics that make it particularly suitable for reconstructive procedures. Despite its wide use in clinic, the structure of this depot has never been studied in detail and it is not known if structural differences exist among trochanteric fat and other subcutaneous WAT depots. The present study was performed on trochanteric fat pad with the aim to clarify the morphology of its adipocytes, stroma and microcirculation, with particular reference to the stem niches. Histological and ultrastructural studies showed that the main peculiar feature of the trochanteric fat concerns its stromal component, which appears less dense than in the other subcutaneous WATs studied. The intra-parenchymal collagen stroma is poor and the extracellular compartment shows large spaces, filled with electron-light material, in which isolated collagen bundles are present. The adipocytes are wrapped in weak and easily detachable collagen baskets. These connective sheaths are very thin compared to the sheaths in other subcutaneous WAT depots. The capillaries are covered by large, long and thin elements surrounded by an external lamina; these perivascular cells are poor in organelles and mainly contain poly-ribosomes. In conclusion, when compared to other WAT deposits, the trochanteric fatty pad shows structural peculiarities in its stroma and microcirculation suggesting a high regenerative potential. Resistance, dissociability, microvascular weft and high regenerative potential make the trochanteric fatty pad a privileged source for harvesting in autologous WAT-based regenerative procedures

Engineered Sleeping Beauty Transposon as Efficient System to Optimize Chimp Adenoviral Production

Sleeping Beauty (SB) is the first DNA transposon employed for efficient transposition in vertebrate cells, opening new applications for genetic engineering and gene therapies. A transposon-based gene delivery system holds the favourable features of non-viral vectors and an attractive safety profile. Here, we employed SB to engineer HEK293 cells for optimizing the production of a chimpanzee Adenovector (chAd) belonging to the Human Mastadenovirus C species. To date, chAd vectors are employed in several clinical settings for infectious diseases, last but not least COVID-19. A robust, efficient and quick viral vector production could advance the clinical application of chAd vectors. To this aim, we firstly swapped the hAd5 E1 with chAd-C E1 gene by using the CRISPR/Cas9 system. We demonstrated that in the absence of human Ad5 E1, chimp Ad-C E1 gene did not support HEK293 survival. To improve chAd-C vector production, we engineered HEK293 cells to stably express the chAd-C precursor terminal protein (ch.pTP), which plays a crucial role in chimpanzee Adenoviral DNA replication. The results indicate that exogenous ch.pTP expression significantly ameliorate the packaging and amplification of recombinant chAd-C vectors thus, the engineered HEK293ch.pTP cells could represent a superior packaging cell line for the production of these vectors

Allele-specific editing ameliorates dominant retinitis pigmentosa in a transgenic mouse model

Retinitis pigmentosa (RP) is a group of progressive retinal degenerations of mostly monogenic inheritance, which cause blindness in about 1:3,500 individuals worldwide. Heterozygous variants in the rhodopsin (RHO) gene are the most common cause of autosomal dominant RP (adRP). Among these, missense variants at C-terminal proline 347, such as p.Pro347Ser, cause severe adRP recurrently in European affected individuals. Here, for the first time, we use CRISPR/Cas9 to selectively target the p.Pro347Ser variant while preserving the wild-type RHO allele in vitro and in a mouse model of adRP. Detailed in vitro, genomic, and biochemical characterization of the rhodopsin C-terminal editing demonstrates a safe downregulation of p.Pro347Ser expression leading to partial recovery of photoreceptor function in a transgenic mouse model treated with adeno-associated viral vectors. This study supports the safety and efficacy of CRISPR/Cas9-mediated allele-specific editing and paves the way for a permanent and precise correction of heterozygous variants in dominantly inherited retinal diseases

Perfil hormonal em mudas Bordô/VR043-43 pós-forçagem a partir de estacas com diferentes épocas de coleta e tempos de armazenagem.

Híbridos Vitis rotundifolia (ex.: VR043-43) têm sido empregados como porta-enxertos em vinhedos com problemas de estresses bióticos, principalmente no estado de Santa Catarina. Entretanto, a produção de mudas é dificultada com estacas lignificadas, pela restrição em calogênese de enxertia e enraizamento desses genótipos.CLAUDIO DE ANDRADE BARROS, CNPUV, 29742

Dark chocolate modulates platelet function with a mechanism mediated by flavan-3-ol metabolites

Cocoa is a rich source bioactive compounds, i.e., flavan-3-ols, and its consumption has been associated with several beneficial effects, such as the positive modulation of the hemostasis targeted by the platelet function. However, these phenolic compounds have a very low bioavailability and extensively undergo phase I and II metabolism, with the appearing into the bloodstream of (epi) catechin conjugates and phenyl-g-valerolactones and their conjugates, at different times. The aims of this study were to explore the effect of dark chocolate on platelet function and to investigate the relationship between this interplay and flavan-3-ol derived metabolites. Eighteen healthy male volunteers ingested 50 g of 90% cocoa chocolate within 5 minutes. Blood samples were collected immediately before chocolate ingestion (T0) and 4 hours afterwards (T1). Platelet function analyzer (PFA)-100 closure time was assessed using collagen/adenosine-50-diphosphate (COL/ADP) and collagen/epinephrine (COL/EPI) cartridges. Plasma flavan-3-ol metabolites were identified and quantified by means of liquid chromatography coupled to a triple quadrupole mass spectrometer (UHPLC-ESI-MS/MS). Results evidenced a significant increase of COL/ADP-induced PFA-100 closure time, but not COL/EPI, 4 hours after ingestion of dark chocolate. Total plasma structurally-related (epi)catechin metabolite (SREM) concentration significantly increased at T1, together with 4 out of the 6 detected metabolites. Total phenyl-g-valerolactone concentrations remained unchanged. Spearman correlations evidenced a strong correlation between COL/ADP closure time and SREMs, mainly led by (epi)catechin-sulfate isomers. These data confirm that the potential beneficial effect of dark chocolate on primary hemostasis may be mediated by flavan-3-ol circulating metabolites

Qualidade de mudas de videira em pós-forçagem submetidas a aplicação de etefom, diferentes épocas de coleta e tempo de câmara fria.

A procura por mudas de videira com qualidade superior tem se intensificado ultimamente, exigindo aprimoramento no processo de produção pelos viveiristas. A cultivar Bordô (B) enxertada nos porta-enxertos P1103 (P) e VR043-43 (VR) são as combinações tradicionais utilizadas no Rio Grande do Sul e em Santa Catarina, respectivamente.Daniel Antunes Souza, 333734, CNPU

Avaliação de duas técnicas de translocação de serrapilheira sobre as assembleias de aranhas (Arachnida: Araneae) e formigas (Hymenoptera: Formicidae)

Due to the current rhythm of ecosystem destruction, the development of new techniques aiming at the conservation of forest remnants and their biota is urgent. Our objective was to compare two techniques of leaf-litter translocation (with and without  withdrawing the pre-extent litter layer), through observations on spider and ant richness and assemblage composition, in order to define the best way to perform translocation. The study was carried out in two Brazilian Atlantic rain forest remnants  (called releaser and receptor fragments) in Salvador, State of Bahia, Brazil. A comparison between the faunas of both fragments was performed before translocation and a significant difference was found both in their richness and composition, which is an essential condition to carry out the experiment. A small scale environmental characterization] was made in order to choose similar units capable of  minimizing the stress on leaf-litter organisms living there. Both for spiders and ants, the species richness increased in units that received leaf-litter and it was higherfor spiders in units where the pre-extent litter had not been removed. For the ants, the two forms of translocation seem to be efficient. Nevertheless, taking into account the two groups of organisms, it is suggested that the translocation must be done maintaining the pre-extent leaf-litter.Key words: conservation, litter, fauna manipulation, spiders, ants.Com o ritmo atual de destruição dos ecossistemas surge a necessidade de se investir em técnicas que visem à conservação dos remanescentes florestais e das suas espécies. Nosso objetivo foi comparar duas técnicas para translocação da serrapilheira (com e sem a retirada da camada de serrapilheira preexistente), observando como varia a riqueza e composição de aranhas e formigas no local da translocação, a fim de definir a melhor forma de realizá-la. O estudo foi realizado em dois remanescentes de Mata Atlântica (fragmento doador e receptor) localizados em Salvador (BA). Antes da translocação foi realizada uma comparação de ambas as faunas, entre os fragmentos, e foi  encontrada diferença significativa tanto na riqueza quanto na composição. Assim pôde-se realizar o experimento. Para tanto, foi realizada a  caracterização ambiental a fim de translocar a serrapilheira para as unidades mais similares na  tentativa de minimizar o estresse dos organismos. A riqueza em espécies tanto de aranhas quanto de  formigas aumentou nas unidades que receberam a serrapilheira, sendo para aranhas mais elevada nas unidades onde não foi retirada a serrapilheira preexistente. Já para as formigas as duas formas parecem eficientes. Contudo, levando em consideração os dois grupos, propõe-se que a translocação deve ser feita mantendo a serrapilheira preexistente.Palavras-chave: conservação, serrapilheira, manipulação da fauna, aranhas, formigas

Zoneamento das condições atuais e futuras de frio para o controle natural da endodormência em clones de Moscato Branco e cvs Vitis labrusca no sul do Brasil.

Como uma planta de clima temperado, a videira necessita de um acúmulo de horas frio (HF, T<7,2°C) para superar o estado de endodormência e ter uma brotação regular, o que é variável entre genótipos. Confrontando essa demanda com as previsões de mudanças climáticas e diminuição de frio, o presente trabalho objetivou contabilizar e delimitar o zoneamento atual e futuro para o cultivo de diferentes clones de Moscato Branco (Vitis vinifera) e cultivares Vitis labrusca

Synchrotron oscillation effects on an rf-solenoid spin resonance

New measurements are reported for the time dependence of the vertical polarization of a 0: 97 GeV/c deuteron beam circulating in a storage ring and perturbed by an rf solenoid. The storage ring is the cooler synchrotron (COSY) located at the Forschungszentrum Julich. The beam polarization was measured continuously using a 1.5 cm thick carbon target located at the edge of the circulating deuteron beam and the scintillators of the EDDA detector. An rf solenoid mounted on the ring was used to generate fields at and near the frequency of the 1 - G gamma spin resonance. Measurements were made of the vertical beam polarization as a function of time with the operation of the rf solenoid in either fixed or continuously variable frequency mode. Using rf-solenoid strengths as large as 2.66 x 10(-5) revolutions/turn, slow oscillations (similar to 1 Hz) were observed in the vertical beam polarization. When the circulating beam was continuously electron cooled, these oscillations completely reversed the polarization and showed no sign of diminishing in amplitude. But for the uncooled beam, the oscillation amplitude was damped to nearly zero within a few seconds. A simple spin-tracking model without the details of the COSY ring lattice was successful in reproducing these oscillations and demonstrating the sensitivity of the damping to the magnitude of the synchrotron motion of the beam particles. The model demonstrates that the characteristic features of measurements made in the presence of large synchrotron oscillations are distinct from the features of such measurements when made off resonance. These data were collected in preparation for a study of the spin coherence time, a beam property that needs to become long to enable a search for an electric dipole moment using a storage ring