SummaryPseudoplatystoma coruscans is a very popular species for tropical fish culture as it has boneless meat of delicate taste and firm texture. Few studies on fish reproductive biology refer to the morphological features of eggs. The goal, therefore, of this present work was to perform a structural and ultrastructural analysis of fertilization and embryonic development in P. coruscans. The incubation period, from fertilization to hatching, lasts 13 h at 28/29• C and 18 h at 27• C. The oocytes had a mean diameter of 0.95 mm and hatched larvae were 2.55 mm in diameter. Analysing their development, we observed round, yellow oocytes that bore a double chorion membrane and a single micropyle. At 10 s after fertilization, several spermatozoa were detected attached to the oocyte surface. After 1 min of development, a fertilization cone that obstructed the micropyle could be observed. Segmentation started between 20 and 30 min after fertilization, when the egg cell was then formed. The first cleavage occurred between 30 and 45 min after fertilization, prior to reaching the morula stage (75 and 90 min after fertilization). The epiboly movement started at 120 and 180 min after fertilization and ended at 360 and 480 min after fertilization. Differentiation between cephalic and caudal region was detected after 420 and 600 min after fertilization and larvae hatched between 780 and 1080 min after fertilization. Seven main embryonic development stages were identified: egg cell, cleavage, morula, blastula, gastrula, segmentation with differentiation between cephalic and caudal regions, and hatching.

Experimental autoimmune encephalomyelitis (EAE) is an animal model to study multiple sclerosis (MS). Considering the tolerogenic effects of active vitamin D, we evaluated the therapeutic effect of myelin oligodendrocyte glycoprotein (MOG) associated with active vitamin D in EAE development. EAE was induced in female C57BL/6 mice by immunization with MOG emulsified with Complete Freund’s Adjuvant plus Mycobacterium tuberculosis. Animals also received two intraperitoneal doses of Bordetella pertussis toxin. One day after immunization, mice were treated with 0,1μg of 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3) every other day during 15 days (on days 1, 3, 5, 7, 9, 11, 13 and 15). MOG (150μg) was co-administered on days 3 and 11. The administration of 1,25(OH) 2D3 or MOG determined significant reduction in EAE incidence and in clinical scores. When MOG was associated with 1,25(OH) 2D3 the animals did not develop EAE. Spleen and central nervous system (CNS) cell cultures from this group produced less IL-6 and IL-17 upon stimulation with MOG in comparison to the EAE control group. In addition, this treatment inhibited dendritic cells maturation in the spleen and reduced inflammatory infiltration in the CNS. The association of MOG with 1,25(OH) 2D3 was able to control EAE development.

In this work, we developed a solid lipid nanoparticle (SLN) formulation with (+)-limonene 1,2-epoxide and glycerol monostearate (Lim-SLNs), stabilized with Poloxamer® 188 in aqueous dispersion to modify the release profile of the loaded monoterpene derivative. We also evaluated the role of SLNs in lipid peroxidation and cytotoxicity in a spontaneously transformed aneuploid immortal keratinocyte cell line from adult human skin (the HaCaT cell line). For the cell viability assay, the colorimetric 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was used. Lim-SLNs with a loading capacity and encapsulation efficiency of 0.39% and 63%, respectively, were produced by high pressure homogenization. A mean particle size of 194 ± 3.4 nm and polydispersity index of 0.244 were recorded for the loaded Lim-SLNs, as compared to 203 ± 1.5 nm (PI 0.213) for the non-loaded (blank) SLNs. The loading of the monoterpene derivative into glycerol monostearate SLNs fitted into the zero-order kinetics, and ameliorated both lipid peroxidation and cytotoxicity in a keratinocyte cell line. A promising formulation for antioxidant and anti-tumoral activities is here proposed.

RESUMO.-[Análise hematológica comparativa de ovelhas Morada Nova e Santa Inês em todos os estágios reprodutivos.] A análise hematológica fornece informação confiável sobre a saúde dos animais. É uma variável importante na avaliação da capacidade adaptativa e produtiva das raças sob condições ambientais desfavoráveis. É uma avaliação que aliada a outros fatores genéticos, ambientais e de manejo pode se tornar cada vez mais útil para a sustentabilidade futura da agricultura convencional em um ambiente quente. A capacidade adaptativa é afetada por alguns fatores como raça, gestação e lactação. It is an important variable in the assessment of adaptive and productive capacity of breeds under unfavorable environmental conditions. It is an assessment that combined with other genetic and environmental factors and management can become useful for the future sustainability of mainstream agriculture in a hot environment. Adaptive capacity is affected by some factors such as breed, pregnancy and lactation. Therefore, hematological profile is an important indicator of animal health and production. The objective of this study was to perform the comparative hematological analysis of Santa Inês and Morada Nova breeds (native ewes from Brazil) in all different reproductive stages. Twenty Santa Ines and 20 Morada Nova sheep distributed in a completely randomized design in a split-plot arrangement over time were used. To obtain blood counts, blood samples were collected by jugular venipuncture every 14 days, always in the morning, before the animals were released into the pasture. The Morada Nova breed had higher hemoglobin and total leukocyte count than Santa Inês breed. Regarding the influence of pregnancy and puerperium on the eritrogram, there was an increment in red blood cell, hemoglobin and packed cell volume of both breeds in middle pregnancy. However, had a reduction in late pregnancy. There were a reestablishment of the blood cell counts during the puerperium period. It was verified an increase on neutrophil: lymphocyte ratio indicated that ewes in the late pregnancy, postpartum and puerperium were under stress conditions compared to others reproductive stages. The total plasma proteins also increased during this period to compensate for the high nutritional requirements of the fetus, and these levels remained high until the end of the puerperal phase, when the lambs were weaned. The pregnancy and puerperium influenced all erythrocyte indices and changed the total leukocyte count. perfil hematológico é um indicador importante da saúde e produção animal. O objetivo deste estudo foi realizar a análise hematológica comparativa de ovelhas Morada Nova e Santa Inês (ovelhas nativas do Brasil) em todos os está-gios reprodutivos. Foram utilizadas 20 ovelhas Santa Inês e 20 Morada Nova distribuídas em um delineamento inteiramente casualizado, em parcelas subdivididas e medidas repetidas no tempo. Para a análise do sangue, foram coletadas amostras por venopunção jugular a cada 14 dias, sempre colhidas pela manhã, antes que os anim...

No abstract

We have developed a new cationic solid lipid nanoparticle (SLN) formulation, composed of Compritol ATO 888, poloxamer 188 and cetyltrimethylammonium bromide (CTAB), to load perillaldehyde 1,2-epoxide, and surface-tailored with a monoclonal antibody for site-specific targeting of human epithelial growth receptor 2 (HER2). Perillaldehyde 1,2-epoxide-loaded cationic SLN (cPa-SLN), with a mean particle size (z-Ave) of 275.31 ± 4.78 nm and polydispersity index (PI) of 0.303 ± 0.081, were produced by high shear homogenization. An encapsulation efficiency of cPa-SLN above 80% was achieved. The release of perillaldehyde 1,2-epoxide from cationic SLN followed the Korsemeyer–Peppas kinetic model, which is typically seen in nanoparticle formulations. The lipid peroxidation of cPa-SLN was assessed by the capacity to produce thiobarbituric acid-reactive substances, while the antioxidant activity was determined by the capacity to scavenge the stable radical DPPH. The surface functionalization of cPa-SLN with the antibody was done via streptavidin-biotin interaction, monitoring z-Ave, PI and ZP of the obtained assembly (cPa-SLN-SAb), as well as its stability in phosphate buffer. The effect of plain cationic SLN (c-SLN, monoterpene free), cPa-SLN and cPa-SLN-SAb onto the MCF-7 cell lines was evaluated in a concentration range from 0.01 to 0.1 mg/mL, confirming that streptavidin adsorption onto cPa-SLN-SAb improved the cell viability in comparison to the cationic cPa-SLN.

Experimental autoimmune encephalomyelitis (EAE) is an artificially induced demyelination of the central nervous system (CNS) that resembles multiple sclerosis in its clinical, histopathological, and immunological features. Activated Th1 and Th17 cells are thought to be the main immunological players during EAE development. This study was designed to evaluate peripheral and local contribution of IL-17 to acute and chronic EAE stages. C57BL/6 mice were immunized with MOG plus complete Freund's adjuvant followed by pertussis toxin. Mice presented an initial acute phase characterized by accentuated weight loss and high clinical score, followed by a partial recovery when the animals reached normal body weight and smaller clinical scores. Spleen cells stimulated with MOG produced significantly higher levels of IFN-γ during the acute period whereas similar IL-17 levels were produced during both disease stages. CNS-infiltrating cells stimulated with MOG produced similar amounts of IFN-γ but, IL-17 was produced only at the acute phase of EAE. The percentage of Foxp3+ Treg cells, at the spleen and CNS, was elevated during both phases. The degree of inflammation was similar at both disease stages. Partial clinical recovery observed during chronic EAE was associated with no IL-17 production and presence of Foxp3+ Treg cells in the CNS.

RESUMO -Este trabalho foi realizado objetivando-se determinar a composição corporal de proteína, gordura e energia e as exigências nutricionais de proteína e energia para mantença e ganho de peso de ovinos Morada Nova. Foram utilizados 30 cordeiros com peso vivo (PV) médio inicial de 15 kg. Para determinação da composição corporal, pela metodologia do abate comparativo, seis cordeiros (animaisreferência) foram abatidos aos 15 kg, seis aos 20 kg (abate intermediário) e os demais foram distribuídos em seis grupos (um para cada dieta) de três animais, de acordo com a relação volumoso(V):concentrado(C): 40:60, 55:45 e 70:30. Os cordeiros em cada grupo foram abatidos quando o que recebia a dieta com maior teor de concentrado atingiu 25 kg de PV. A composição corporal variou de 181,53 a 178,74 g de proteína; 72,37 a 131,11 g de gordura e 1,81 a 2,34 Mcal de energia por kg de peso de corpo vazio (PCV). As exigências líquidas de ganho variaram de 222,30 a 218,6 g de proteína e de 3,30 a 4,28 Mcal de energia por kg de PV ganho. As perdas endógenas de nitrogênio foram de 0,332 g/kg 0,75 /dia. A exigência líquida de energia para mantença, estimada pela produção de calor em jejum, foi de 52,49 kcal/ kg 0,75 de PCV. As exigências de proteína e energia metabolizável para cordeiros dos 15 aos 25 kg de PV, com ganho de peso diário de 100 g, oscilaram de 53,46 a 60,19 g/dia e de 1,47 a 2,00 Mcal/dia, respectivamente.Palavras-chave: exigências de ganho, mantença, nitrogênio, perdas endógenas, produção de calor, ovinos Body Composition and Nutritional Requirements of Protein and Energy for Morada Nova LambsABSTRACT -This investigation was carried out to determine body composition of protein, fat, and energy as well as nutritional requirements of protein and energy for maintenance and weight gain of Morada Nova Lambs. Thirty lambs averaging 15 kg of initial body weight (BW) were used in this trial. To determine the body composition using the comparative slaughter approach six lambs were slaughtered at 15 kg (reference animals), six at 20 kg (intermediary slaughter), and the remaining were distributed in one of six groups of three animals (one animal per diet) receiving the following forage (F):concentrate(C) ratios: 1) 40F:60C; 2) 55F:45C, and 3) 70F:30C. Lambs in each group were slaughtered when the animal fed the highest amount of concentrate reached 25 kg of BW. Body composition varied from 181.53 to 178.74 g of protein, 72.37 to 131.11 g of fat, and 1.81 to 2.34 Mcal of energy per kg of empty body weight (EBW). The net requirements of gain varied from 222.30 to 218.6 g of protein and from 3.30 to 4.28 Mcal of energy per kg of gained BW. The endogenous losses of nitrogen were of 0.332 g/kg 0.75 /day. The net requirement of energy for maintenance, estimated by heat production in fasted animals, was of 52.49 Kcal/kg 0.75 of EBW. The requirements of metabolizable protein and energy for lambs from 15 to 25 kg of BW with daily weight gain of 100 g varied from 53.46 to 60.19 g/day and from 1.47 a 2.00 Mcal/day, respectively.