Quantitative analysis of receptor-mediated uptake and pro-apoptotic activity of mistletoe lectin-1 by high content imaging.

Ribosome inactivating proteins (RIPs) are highly potent cytotoxins that have potential as anticancer therapeutics. Mistletoe lectin 1 (ML1) is a heterodimeric cytotoxic protein isolated from European Mistletoe and belongs to RIP class II. The aim of this project was to systematically study ML1 cell binding, endocytosis pathway(s), subcellular processing and apoptosis activation. For this purpose, state of the art cell imaging equipment and automated image analysis algorithms were used. ML1 displayed very fast binding to sugar residues on the membrane and energy-dependent uptake in CT26 cells. The co-staining with specific antibodies and uptake blocking experiments revealed involvement of both clathrin-dependent and -independent pathways in ML1 endocytosis. Co-localization studies demonstrated the toxin transport from early endocytic vesicles to Golgi network; a retrograde road to the endoplasmic reticulum. The pro-apoptotic and antiproliferative activity of ML1 were shown in time lapse movies and subsequently quantified. ML1 cytotoxicity was less affected in multidrug resistant tumor cell line 4T1 in contrast to commonly used chemotherapeutic drug (ML1 resistance index 6.9 vs 13.4 for doxorubicin; IC50: ML1 1.4 ng/ml vs doxorubicin 24000 ng/ml). This opens new opportunities for the use of ML1 as an alternative treatment in multidrug resistant cancers.

Valores nutricionais do milho de diferentes qualidades para frangas de reposição na fase de recria / Nutritional value of corn with different qualities for egg pullets in the growing phase

O experimento objetivou determinar os valores nutricionais de diferentes frações de milho obtidas por meio de estratificação em mesa densimétrica na recria de frangas. Os milhos foram designados como: MDA - milho de densidade alta; MDI - milho de densidade intermediária; MDB - milho de densidade baixa; MDT - milho de densidade total, composto de 30% de MDA, 60% de MDI e 10% de MDB. Para a determinação da energia metabolizável corrigida (EMAn) foi utilizado o método de coleta total de excretas em frangas Hy Line de 15 semanas. Os valores de EMAn (kcal/kg na MN) foram: 3.467; 3.340; 3.217 e 3.385kcal/kg e densidade (kg/m³): 818,61; 698,13; 681,80 e 736,39kg/m³ para MDA; MDI; MDB e MDT, respectivamente. O MDB apresentou maior valor em todos os aminoácidos digestíveis, com maior intensidade para o triptofano. As frações de milho foram variáveis quanto ao EMAn e perfil de aminoácidos digestíveis, indicando a necessidade de correções nutricionais para a formulação de rações de custo mínimo...

The experiment aimed to determine the nutritional value of different corn fractions obtained by stratification in a gravity table of replacement pullets. The corn was designated as MDA - high density corn; MDI - medium density corn; MDB - low density corn; MDT - total corn density, composed of 30% MDA, 60% MDI and 10% MDB. To determine the corrected metabolizable energy (AME N) a method for total collection of excreta with pullets Hy Line was used for 15 weeks. AME N (kcal/kg in MN) were: 3467, 3340, 3217 and 3385kcal/kg and density (kg/m³): 818.61, 698.13, 681.80 and 736.39 for MDA, MDI, MDB and MDT, respectively. The MDB showed the highest value in all the digestible amino acids, with higher intensity for the tryptophan. The fractions of corn were variable as to AME N and digestible amino acid profiles, indicating the need for corrections to the nutritional feed formulation of minimum cost...

Dexamethasone-loaded poly(ε-caprolactone)/silica nanoparticles composites prepared by supercritical CO2 foaming/mixing and deposition.

A supercritical carbon dioxide (scCO2)-assisted foaming/mixing method (SFM) was implemented for preparing dexamethasone (DXMT)-loaded poly(ε-caprolactone)/silica nanoparticles (PCL/SNPs) composite materials suitable for bone regeneration. The composites were prepared from PCL and mesoporous SNPs (MCM-41/SBA-15) by means of scCO2-assisted SFM at several operational pressures, processing times and depressurization conditions. DXMT was loaded into SNPs (applying a scCO2 solvent impregnation/deposition method - SSID) and into PCL/SNPs composites (using the SFM method). The effects of the employed operational and compositional variables on the physicochemical and morphological features as well as in the in vitro release profiles of DXMT were analyzed in detail. This work demonstrates that the above-referred scCO2-based methods can be very useful for the preparation of DXMT-loaded PCL/SNPs composites with tunable physicochemical, thermomechanical, morphological and drug release properties and suitable for hard-tissue regeneration applications.

The impact of maternal post-partum depression on the language development of children at 12 months.

BACKGROUND: Language is one of the most important acquisitions made during childhood. Before verbal language, a child develops a range of skills and behaviours that allow the child to acquire all communication skills. Factors such as environmental factors, socio-economic status and interaction with parents can affect the acquisition of vocabulary in children. Post-partum depression can negatively affect the first interactions with the child and, consequently, the emotional, social and cognitive development of the child. OBJECTIVE: To analyse the effect of the duration of the mother's depression on the language development of children at 12 months old. METHODS: This was a longitudinal study. The participants of this study were mothers who had received prenatal care from the Brazilian National System of Public Health in Pelotas city, State of Rio Grande do Sul, Brazil. The mothers were interviewed at two different time points: from 30 to 90 days after delivery and at 12 months after delivery; the children were also evaluated at this later time point. To diagnose maternal depression, we used the Mini International Neuropsychiatric Interview, and to assess child development, we used the language scale of the Bayley Scales of Infant Development III. RESULTS: We followed 296 dyads. Maternal depression at both time points (post partum and at 12 months) was significantly associated with the language development of infants at 12 months of age. This impact was accentuated when related to the duration of the disorder. Older women and women with more than two children were more likely to have children with poorer language development, while women who were the primary caregiver had children with higher scores on the language test. CONCLUSION: The findings indicate that maternal age, parity, primary caregiver status and duration of post-partum depression are associated with the language development of the child.

Balanço eletrolítico da dieta e desempenho de frangos em condições naturais de estresse calórico / Electrolyte balance of diets under performance of broiler in naturals conditions of heat stress

Avaliou-se o efeito da suplementação de eletrólitos sobre o desempenho de frangos, utilizando bicarbonato de sódio (NaHCO3) e cloreto de potássio (KCl) para ajuste do balanço eletrolítico da dieta (BED) com duas porcentagens de proteína bruta (PB). Utilizaram-se 800 frangos machos, alojados em 20 boxes, distribuídos em delineamento inteiramente ao acaso, com quatro tratamentos, cinco repetições e 40 aves por unidade experimental. Os tratamentos foram: dieta basal; dieta sem redução de PB suplementada com eletrólitos para ajuste do BED para 250mEq/kg; dieta com redução de PB, sem suplementação de eletrólitos; e dieta com redução de PB, com eletrólitos para ajuste do BED para 250mEq/kg. Foram avaliados: consumo da dieta, peso final, ganho de peso, ganho de peso médio diário, conversão alimentar e calórica e eficiência de utilização de nitrogênio e lisina nas fases de sete a 21 e de sete a 42 dias. Não se observaram diferenças (P>0,05) no consumo da dieta. A correção do BED com menor teor de PB melhorou (P<0,05) o peso final, na fase inicial. Houve piora (P<0,05) no ganho de peso e na conversão alimentar nas duas fases. A redução de três pontos percentuais na PB da dieta piorou o desempenho dos frangos.

The effect of the supplementation of electrolytes on de performance of broilers using sodium bicarbonate (NaHCO3) and potassium chloride (KC1) to adjust the electrolytic balance of the diet (BED) with two percentages of raw protein (PB) was studied. 800 male broiler chickens, housed in 20 boxes, distributed in random lineation, with four treatments, five repetitions and 40 birds per experimental unit were used. The treatments were: base diet; diet with no reduction of PB supplemented with electrolytes to adjust BED to 250mEq/kg; diet with a reduction of PB, with no electrolyte supplementation; and diet with the reduction of PB, with electrolytes to adjust the BED to 250mEq/kg. What was evaluated was: diet consumption, final weight, weight gain, average daily weight gain, food conversion and caloric efficiency of the use of nitrogen and lysine in phases from 7 to 21 and 7 to 42 days. No differences were observed (P>0.05) in diet consumption. The correlation of BED with the lower percentage of PB improved (P<0.05) the final weight in the initial phase. Weight gain and food conversion worsened (P<0.05) in both phases. The reduction of three percentage points in the PB in the diet worsened the broiler chicken's performance.