Efeito de diferentes fontes de carboidratos não-fibrosos sobre o pH ruminal e digestibilidade in vitro de forragens tropicais

O presente estudo teve como objetivo avaliar os carboidratos não-fibrosos (CNF) em dietas à base de cana-de-açúcar sobre o pH ruminal e digestibilidade da forragem, e descrever as curvas de degradação dos CNF. O estudo foi composto de dois ensaios. No primeiro, três novilhos canulados no rúmen, com peso vivo de 350 ± 15 kg (Média ± DP), foram alocados em um quadrado latino (QL) 3×3, e alimentados com dietas contendo: milho moído (MM, tamanho de partículas 0,9 mm), laminado a vapor (MLV) ou polpa cítrica peletizada (PCP). Cada período tinha 14 d, sendo os primeiros 12 para adaptação e o 13º para a medição seriada do pH e o 14º para a coleta de líquido ruminal e incubação in vitro para digestibilidade da MS e FDN (DIVMS e DIVFDN) de feno de bermudagrass (Feno) e silagens de milho (SM) e cana (SC). No segundo ensaio, coletou-se fluido ruminal de um touro canulado, alimentado com silagem de milho e concentrado padrão, para digestão in vitro dos CNF em vários tempos. Esses resultados foram utilizados para ajustar as curvas de degradação dos CNF e calcular o tempo de colonização, frações alimentares e taxa de degradação. Os resultados do primeiro ensaio foram analisados em um QL 3×3. O modelo dos parâmetros de digestibilidade incluiu efeito fixo de forragem (Alimento), dieta com CNF (Dieta) e interação (Alimento × Dieta), e efeito aleatório de animal e período. O modelo para pH incluiu efeito fixo de Dieta, Tempo como medida repetida, animal e período como aleatórios. Foi considerada a probabilidade significativa de ≤ 5% (α = 0,05). As curvas de degradação dos CNF foram ajustadas pelo PROC NLIN do SAS, e parâmetros de equação comparados por intervalo de confiança. Houve interação Dieta × Tempo no pH ruminal (P = 0,04), onde o MLV diminuiu o pH comparado com PCP e MM apenas no tempo 6 h. Não houve interação Alimento×Dieta (P > 0,05) para nenhum parâmetro de digestibilidade. Houve efeito de Alimento sobre a DIVMS e DIVFDN, após 30 e 48 h de incubação (P < 0,01). A SM teve a maior DIVMS, seguido por SC e Feno, após 30 e 48 h de incubação. A SM teve a maior DIVFDN após 30 h, comparado com SC e Feno. No entanto, para DIVFDN após 48 h, a SM teve maior média, seguida da SC e Feno. O fluido ruminal de animais alimentados com MLV diminuiu a DIVMS e DIVFDN (P < 0.05) de todas as forragens, após 48 h. Resultados do segundo ensaio mostram que PCP diminuiu o tempo de colonização, fração B e aumentou a kd comparado com os dois milhos, e MLV apresentou maior kd que o MM. Em conclusão, a dieta com MLV diminuiu o pH ruminal no tempo 6 h e, consequentemente, diminuiu a DIVFDN das forragens avaliadas. Embora PCP tenha apresentado menor tempo de colonização e maior taxa de degradação da fração B, não afetou negativamente o pH do rúmen nem a digestibilidade da fibra das forragens.The present study aimed to evaluate non-fiber carbohydrates (NFC) in sugarcane-based diets on rumen pH, and forage digestibility, and to describe NFC degradation curves. The study consisted of two trials. For the first trial, three rumen cannulated steers, BW of 350 ± 15 kg (mean ± SE), were assigned in a 3×3 Latin Square (LS) design. They were fed diets containing finely-ground (0.9 mm average particle size) corn (GC), steam-rolled corn (SRC), or pelleted citrus pulp (PCP). Each period had 14 d, with the first 12 for adaptation. The 13th d was for serial measurement of rumen pH, and the14th for rumen fluid collection and in vitro incubation for DM and NDF digestibility (IVDMD and IVNDFD) of bermudagrass hay (Hay), corn (CS), and sugarcane (SS) silages. In the second trial, rumen fluid of a cannulated bull, fed corn silage and a regular concentrate, was collected for in vitro digestion of NFC for multiple time points. The incubation results were used to adjust the NFC degradation curves, and calculate lag-time, feed fractions, and degradation rate. Data from first trial was analyzed in a 3×3 LS. The model for the digestibility parameters included fixed effects of forage (Feed), diets with NFC (Diet), and their interaction (Feed × Diet), and random effect of animal and period. The model for rumen pH included fixed effect of diet, time as repeated measures, animal and period as random effects. The significance was considered at probability ≤ 5% (α = 0.05). The NFC degradation curves were adjusted using the PROC NLIN procedure from SAS, and equation parameters compared using confidence intervals. There was a Diet × Time interaction on rumen pH (P = 0.04), where SRC decreased pH compared to PCP and GC diets at the time 6 h, only. There was no Feed × Diet interaction effect (P > 0.05) for any digestibility parameter. There was a Feed effect on both IVDMD and IVNDFD, either after 30 or 48 h incubation (P < 0.01). The CS had the greatest IVDMD, followed by SS and Hay, after 30 and 48 h of incubation. The CS had the greatest IVNDFD after 30 h, compared to SS and Hay. However, for IVNDFD after 48 h, CS presented the greatest mean, followed by SS and Hay. The rumen fluid from animals fed SRC decreased both IVDMD and IVNDFD (P < 0.05) of all roughages after 48 h. Results from the second trial showed that the PCP had lower Lag Time, B fraction and greater kd compared to both corn sources, and SRC had greater kd than GC. In conclusion, the SRC diet decreased rumen pH 6 h after feeding and, consequently, decreased fiber digestibility of the tropical forage sources evaluated. Although the PCP had lower lag time, and faster rate of degradation of B fraction, it did not negatively affect rumen pH or fiber digestibility of forage

Anteproyecto de construccion de una embarcacion tipica de Chiloe

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The Retroviral Restriction Ability of SAMHD1, but Not Its Deoxynucleotide Triphosphohydrolase Activity, Is Regulated by Phosphorylation

SummarySAMHD1 is a cellular enzyme that depletes intracellular deoxynucleoside triphosphates (dNTPs) and inhibits the ability of retroviruses, notably HIV-1, to infect myeloid cells. Although SAMHD1 is expressed in both cycling and noncycling cells, the antiviral activity of SAMHD1 is limited to noncycling cells. We determined that SAMHD1 is phosphorylated on residue T592 in cycling cells but that this phosphorylation is lost when cells are in a noncycling state. Reverse genetic experiments revealed that SAMHD1 phosphorylated on residue T592 is unable to block retroviral infection, but this modification does not affect the ability of SAMHD1 to decrease cellular dNTP levels. SAMHD1 contains a target motif for cyclin-dependent kinase 1 (cdk1) (592TPQK595), and cdk1 activity is required for SAMHD1 phosphorylation. Collectively, these findings indicate that phosphorylation modulates the ability of SAMHD1 to block retroviral infection without affecting its ability to decrease cellular dNTP levels

Oxidative Damage in Lymphocytes of Copper Smelter Workers Correlated to Higher Levels of Excreted Arsenic

Arsenic has been associated with multiple harmful effects at the cellular level. Indirectly these defects could be related to impairment of the integrity of the immune system, in particular in lymphoid population. To characterize the effect of Arsenic on redox status on this population, copper smelter workers and arsenic unexposed donors were recruited for this study. We analyzed urine samples and lymphocyte enriched fractions from donors to determinate arsenic levels and lymphocyte proliferation. Moreover, we studied the presence of oxidative markers MDA, vitamin E and SOD activity in donor plasma. Here we demonstrated that in human beings exposed to high arsenic concentrations, lymphocyte MDA and arsenic urinary levels showed a positive correlation with SOD activity, and a negative correlation with vitamin E serum levels. Strikingly, lymphocytes from the arsenic exposed population respond to a polyclonal stimulator, phytohemaglutinin, with higher rates of thymidine incorporation than lymphocytes of a control population. As well, similar in vitro responses to arsenic were observed using a T cell line. Our results suggest that chronic human exposure to arsenic induces oxidative damage in lymphocytes and could be considered more relevant than evaluation of T cell surveillance

Optimization of universal allogeneic CAR-T cells combining CRISPR and transposon-based technologies for treatment of acute myeloid leukemia

Despite the potential of CAR-T therapies for hematological malignancies, their efficacy in patients with relapse and refractory Acute Myeloid Leukemia has been limited. The aim of our study has been to develop and manufacture a CAR-T cell product that addresses some of the current limitations. We initially compared the phenotype of T cells from AML patients and healthy young and elderly controls. This analysis showed that T cells from AML patients displayed a predominantly effector phenotype, with increased expression of activation (CD69 and HLA-DR) and exhaustion markers (PD1 and LAG3), in contrast to the enriched memory phenotype observed in healthy donors. This differentiated and more exhausted phenotype was also observed, and corroborated by transcriptomic analyses, in CAR-T cells from AML patients engineered with an optimized CAR construct targeting CD33, resulting in a decreased in vivo antitumoral efficacy evaluated in xenograft AML models. To overcome some of these limitations we have combined CRISPR-based genome editing technologies with virus-free gene-transfer strategies using Sleeping Beauty transposons, to generate CAR-T cells depleted of HLA-I and TCR complexes (HLA-IKO/TCRKO CAR-T cells) for allogeneic approaches. Our optimized protocol allows one-step generation of edited CAR-T cells that show a similar phenotypic profile to non-edited CAR-T cells, with equivalent in vitro and in vivo antitumoral efficacy. Moreover, genomic analysis of edited CAR-T cells revealed a safe integration profile of the vector, with no preferences for specific genomic regions, with highly specific editing of the HLA-I and TCR, without significant off-target sites. Finally, the production of edited CAR-T cells at a larger scale allowed the generation and selection of enough HLA-IKO/TCRKO CAR-T cells that would be compatible with clinical applications. In summary, our results demonstrate that CAR-T cells from AML patients, although functional, present phenotypic and functional features that could compromise their antitumoral efficacy, compared to CAR-T cells from healthy donors. The combination of CRISPR technologies with transposon-based delivery strategies allows the generation of HLA-IKO/TCRKO CAR-T cells, compatible with allogeneic approaches, that would represent a promising option for AML treatment

Removing Systemic Barriers to Equity, Diversity, and Inclusion: Report of the 2019 Plant Science Research Network Workshop “Inclusivity in the Plant Sciences”

A future in which scientific discoveries are valued and trusted by the general public cannot be achieved without greater inclusion and participation of diverse communities. To envision a path towards this future, in January 2019 a diverse group of researchers, educators, students, and administrators gathered to hear and share personal perspectives on equity, diversity, and inclusion (EDI) in the plant sciences. From these broad perspectives, the group developed strategies and identified tactics to facilitate and support EDI within and beyond the plant science community. The workshop leveraged scenario planning and the richness of its participants to develop recommendations aimed at promoting systemic change at the institutional level through the actions of scientific societies, universities, and individuals and through new funding models to support research and training. While these initiatives were formulated specifically for the plant science community, they can also serve as a model to advance EDI in other disciplines. The proposed actions are thematically broad, integrating into discovery, applied and translational science, requiring and embracing multidisciplinarity, and giving voice to previously unheard perspectives. We offer a vision of barrier-free access to participation in science, and a plant science community that reflects the diversity of our rapidly changing nation, and supports and invests in the training and well-being of all its members. The relevance and robustness of our recommendations has been tested by dramatic and global events since the workshop. The time to act upon them is now