Characterization of protein hydrolysates of cosmetic use by CE-MS

Protein hydrolysates have been used as active principles in cosmetic products conferring different properties to the final formulations, which are mostly controlled by the peptide size and its amino acid sequence. In this work, capillary electrophoresis coupled to mass spectrometry analyses were carried out in order to investigate such characteristics of protein hydrolysates. Samples of different origins (milk, soy and rice) were obtained from a local company, and were analyzed without a previous preparation step. The background electrolyte (BGE) and sheath liquid compositions were optimized for each sample. The best BGE composition (860 mmol/L formic acid - pH 1.8 - in 70: 30 v/v water/methanol hydro-organic solvent) was chosen based on the overall peak resolution whereas the best sheath liquid was selected based on increased sensitivity and presented different compositions to each sample (10.9-217 mmol/L formic acid in 75: 25-25: 75 v/v water/methanol hydro-organic solvent). Most of the putative peptides in the hydrolysate samples under investigation presented molecular masses of 1000 Da or less. De novo sequencing was carried out for some of the analytes, revealing the hydrophobicity/polarity of the peptides. Hence, the technique has proved to be an advantageous tool for the quality control of industrial protein hydrolysates.

Effect of heat stress or lipopolysaccharide (E-coli) injection on HSP70 levels in the liver and brain of adrenalectomized rats

Hsp70 content (ng Hsp70 mu g total protein(-1)) in the liver and brain of control and adrenalectomized male rats was investigated by Western Blotting after heat stress (40 degrees C) or endotoxin-induced fever (E. coli lipopolysaccharide injection). The increase in rectal temperature was higher after heat stress than after LPS injection, Heat stress affected Hsp70 content of the liver, but not of the brain; however adrenalectomy did not influence any parameter. These results suggest that, under these circumstances, there is no relationship between the hypothalamic-pituitary-adrenal axis and Hsp70 synthesis in liver and brain. (C) 2000 Elsevier B.V. Ltd. All rights reserved.

Performance and carcass characteristics of broiler chickens with different growth potential and submitted to heat stress

In order to evaluate the effects of broiler genotype and of heat exposure on performance, carcass characteristics, and protein and fat accretion, six hundred one-day-old male broilers were randomly assigned in a 2 x 3 factorial arrangement, according to the following factors: genetic group (selected and non-selected broilers) and pair-feeding scheme (Ad(32) - reared under heat stress and fed ad libitum; Ad(23) - reared at thermoneutrality and fed ad libitum; Pf(23) - reared at thermoneutrality and pair fed with Ad(32)), with a total of six treatments with four replicates of 25 birds each. Independent of pair-feeding scheme, selected broilers showed better feed conversion, higher carcass yield, and lower abdominal fat deposition rate. However, as compared to non-selected broilers, they reduced more intensively feed intake when heat exposed, which promoted significant breast-yield decrease, and more pronounced changes on carcass chemical composition. These findings allows concluding that, in both genetic groups, both environmental temperature and feed-intake restriction influence abdominal fat deposition rate and other carcass characteristics; however, the impact of heat exposure on broiler performance is more noticeable on the selected line.

Heat or cold chronic stress affects organ weights and Hsp70 levels in chicken embryos

This study was carried out with the objective of evaluating the effect of heat (38.8 degreesC) or cold (35.8 degreesC) stress on chicken embryo development and tissues Hsp70 levels, after the 13th day of incubation. Embryo weight (percent egg weight), organ weight (percent embryo weight) and Hsp70 levels (ng Hsp70 mug(-1) total protein) in different tissues (liver, breast muscle, heart, lungs, brain and kidney) were studied at the end of incubation. Cold stress induced a lower embryo weight and lower kidney and lungs weights, whereas heart and liver were lighter in heat-stressed embryos. An interaction between temperature and age was obtained only for Hsp70 levels in kidney and heart. Cold-stressed embryos showed higher Hsp70 levels in the brain, lungs and liver; a decrease in brain and breast muscle Hsp70 levels was seen from the 19th to 20th days in control embryos. Hsp70 levels increased with age in kidneys of control embryos and in heart of heat- and cold-stressed embryos. In conclusion, this study showed that chicken embryo organ weights are affected by incubation temperature, and that Hsp70 expression is tissue dependent (higher levels being seen in the brain) being cold-stress more effective in increasing Hsp70 levels in most studied tissues.

Determination of crude protein, crude fiber and crude fat in twenty-five genotypes of kale (B-oleracea var. acephala)

The aim of the study was to determine the percentage of crude protein, crude fiber and crude fat (ether extract) of 25 genotypes of kale from the Germplasm Bank of Instituto Agronomico de Campinas and of one genotype grown in the region of Jaboticabal-SP. The plants were cultivated in the field, and the leaves after collection were pre-dried in a convection oven at 65 degrees C for 72 h. Afterward, the leaves were analyzed for crude protein, crude fiber and crude fat (ether-soluble materials). Significant differences were detected among the different genotypes for all the characteristics examined. of the genotypes studied, six showed more than 30% crude protein: HS-20 (32.56%), Comum (31.70%), Couve de Arthur Nogueira 2 (31.16%), Pires 2 de Campinas (30.63%), Manteiga 1-916 (30.36%), and Manteiga de Ribeirao Pires I-2446 (30.03%). In relation to crude fiber, the highest percentage was seen in the genotype Manteiga de Mococa (10.92%), differing significantly from the other genotypes studied. With regard to crude fat, the highest percentage was found in the genotype HS-20 (3.72%), and Pires 1 de Campinas (3.34%). of the genotypes tested, HS-20 stood out among the others, showing both the highest percentage of protein and fat.

Effects of energy and protein levels on egg quality and performance of laying hens at early second production cycle

One hundred sixty-two commercial 70-wk-old ISA Brown laying hens, previously subjected to induced molting by feed restriction, were distributed in a completely randomized design with 3 x 3 factorial arrangement (i.e., 3 metabolizable energy levels: 2,850; 2,950, and 3,050 kcal of ME/kg) and 3 protein levels (16, 18, and 20% CP), which totaled 9 treatments with 3 replicates of 6 birds each. Experimental diets were offered to birds after the feed restriction period. Performance and egg quality parameters were evaluated in 14-d intervals from the 4th to 12th weeks after forced molting for a total of 4 evaluation periods. Increases in dietary energy and protein levels did not improve performance or egg quality. The levels of 2,850 kcal of ME and 16% protein were sufficient for laying hens starting the second production cycle without decreasing their performance or egg quality.

Composição corporal e exigências nutricionais de proteína e energia para cordeiros Morada Nova

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 (animais-referência) 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/kg0,75/dia. A exigência líquida de energia para mantença, estimada pela produção de calor em jejum, foi de 52,49 kcal/kg0,75 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.

Estudo da reidratação do feijão verde (Vigna unguiculata L. Walp) desidratado por micro-ondas com e sem pré-tratamento osmótico

The green bean has organoleptic and nutritional characteristics that make it an important food source in tropical regions such as the Northeast of Brazil. It is a cheap source of protein and important for nutrition of rural population contributing significantly in subsistence farming of the families from Brazil s northeast. It is consumed in entire region and together with the dry meat and other products composes the menu of typical restaurants, being characterized as an important product for economy of Northeast. The green bean is consumed freshly harvested and has short cycle, being characterized as a very perishable food, which hampers your market. The drying method is an alternative to increase the lifetime and provide a reduction volume of this product making easier your transportation and storage. However is necessary to search ways of drying which keep the product quality not only from the nutritional standpoint but also organoleptic. Some characteristics may change with the drying process such as the coloring, the rehydration capacity and the grains cooking time. The decrease of drying time or of exposure of the grains to high temperature minimizes the effects related with the product quality loss. Among the techniques used to reduce the drying time and improve some characteristics of the product, stands out the osmotic dehydration, widely used in combined processes such as the pretreatment in drying food. Currently the use of the microwaves has been considered an alternative for drying food. The microwave energy generates heat inside of materials processed and the heating is practically instantaneous, resulting in shorter processing times and product quality higher to that obtained by conventional methods. Considering the importance of the green beans for the Northeast region, the wastefulness of production due to seasonality of the crop and your high perishability, the proposal of this thesis is the study of drying grain by microwaves with and without osmotic pretreatment, focusing on the search of conditions of processes which favor the rehydration of the product preserving your organoleptic characteristics. Based on the analysis of the results of osmotic dehydration and dielectric properties was defined the operating condition to be used in pretreatment of the green bean, with osmotic concentration in saline solution containing 12,5% of sodium chloride, at 40°C for 20 minutes. The drying of green bean by microwave was performed with and without osmotic pretreatment on the optimized condition. The osmotic predehydration favored the additional drying, reducing the process time. The rehydration of dehydrated green bean with and without osmotic pretreatment was accomplished in different temperature conditions and immersion time according to a factorial design 22, with 3 repetitions at the central point. According to results the better condition was obtained with the osmotically pretreated bean and rehydrated at a temperature of 60°C for 90 minutes. Sensory analysis was performed comparing the sample of the green bean in natura and rehydrated in optimized conditions, with and without osmotic pretreatment. All samples showed a good acceptance rate regarding the analyzed attributes (appearance, texture, color, odor and taste), with all values above 70%. Is possible conclude that the drying of green bean by microwave with osmotic pretreatment is feasible both in respect to technical aspects and rehydration rates and sensory quality of the product

Nutritional Responses of Rats to Diets Based on Chickpea (Cicer arietinum L.) Seed Meal or Its Protein Fractions

The aim of this study was to isolate the protein fractions from chickpea, var. IAC-Marrocos, as well as to evaluate its in vivo nutritional protein quality. Among the proteins, albumins showed better nutritional value in the in vivo assays and amino acid contents, despite their higher trypsin inhibitor contents. Trypsin inhibitors were found to be heat labile in all samples, but the digestibility results for unheated and heated flour and albumins suggest that their contents are not very decisive. The PER values for casein (not supplemented) were very similar to those of heated flour and unheated or heated albumin and total globulins. The albumin and glutelin fractions showed the best results for PDCAAS, however, lower than those of casein. Despite the high digestibility of the globulin the very low essential amino acid content lowered its PDCAAS, and it had the lowest values.

Reduced pancreatic β-cell mass is associated with decreased FoxO1 and Erk1/2 protein phosphorylation in low-protein malnourished rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Interleukin-6 and procalcitonin in children with sepsis and septic shock

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Amino acid availability and protein digestibility of several protein sources for Nile tilapia, Oreochromis niloticus

Apparent amino acid availability coefficients and protein digestibility of four animal products [fish meal (FM), meat and bone meal (MBM), poultry by-product and feather meal] and four plant protein-rich products [soybean meal (SBM), cottonseed meal-28, cottonseed meal-38 and corn gluten meal (CGM)] were determined for Nile tilapia, Oreochromis niloticus. Ingredients were incorporated to a practical reference diet at a 7 : 3 ratio (70% of reference diet and 30% of test ingredient). Chromic oxide was used as external digestibility marker. Among animal products poultry by-product meal (PBM; 89.7%) and FM (88.6%) presented the highest apparent protein digestibility (APD) while MBM (78.4%) and feather meal (78.5%) presented the lowest APD. Among plant protein-rich products CGM (91.4%) and SBM (92.4%) presented the highest APD values while cottonseed meal-28 presented the lowest APD (78.6%). Average apparent amino acid availability of feed ingredients was similar to protein digestibility with 92.3%, 89.6%, 73.4%, 80.7%, 88.9%, 84.4%, 91.2% and 79.7% values for SBM, CGM, cottonseed meal-28 and 38, FM, MBM, PBM and feather meal respectively. These results indicate that O. niloticus is able to utilize efficiently different feedstuffs.

Identification of the SP22 Sperm Protein in Santa Ines and Dorper Rams

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Actinobacillus actinomycetemcomitans lipopolysaccharide induces interleukin-6 expression through multiple mitogen-activated protein kinase pathways in periodontal ligament fibroblasts

Actinobacillus actinomycetemcomitans plays a major role in the pathogenesis of aggressive periodontitis. Lipopolysaccharide (LPS) derived from A. actinomycetemcomitans is a key factor in inflammatory cytokine generation within periodontal tissues. In this study, we identify major mitogen-activated protein kinase (MAPK) signaling pathways induced by A. actinomycetemcomitans LPS, Escherichia coli LPS and interleukin-1 beta (IL-1 beta) in a murine periodontal ligament (mPDL) fibroblast cell line. Immunoblot analysis was used to assess the phosphorylated forms of p38, extracellular-regulated kinase (ERK) and c-jun N-terminal kinase (JNK) MAPK following stimulation with A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta. IL-6 mRNA induction was detected via reverse transcription-polymerase chain reaction, while protein levels were quantified via enzyme-linked immunosorbent assays (ELISA). We utilized biochemical inhibitors of p38, ERK and JNK MAPK to identify the MAPK signaling pathways needed for IL-6 expression. Additional use of stable mPDL cell lines containing dominant negative mutant constructs of MAPK kinase-3 and -6 (MKK-3/6) and p38 null mutant mouse embryonic fibroblast (MEF) cells were used to substantiate the biochemical inhibitor data. Blocking p38 MAPK with SB203580 reduced the induction of IL-6 mRNA by A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta by > 70%, > 95% and similar to 60%, respectively. IL-6 ELISA indicated that blocking p38 MAPK reduced the IL-6 protein levels induced by A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta by similar to 60%, similar to 50% and similar to 70%, respectively. All MAPK inhibitors significantly reduced the IL-6 protein levels induced by A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta whereas only p38 inhibitors consistently reduced the A. actinomycetemcomitans LPS, E. coli LPS and IL-1 beta induction of IL-6 mRNA steady-state levels. The contribution of p38 MAPK LPS-induced IL-6 expression was confirmed using MKK-3/6 dominant negative stable mPDL cell lines. Wild-type and p38 alpha(-/-) MEF cells provided additional evidence to support the role of p38 alpha MAPK in A. actinomycetemcomitans LPS-stimulated IL-6. Our results indicate that induction of IL-6 by E. coli LPS, IL-1 beta and A. actinomycetemcomitans LPS requires signaling through MKK-3-p38 alpha ERK, JNK and p38 MAPK in mPDL cells.

Growth performance of Nile tilapia, Oreochromis niloticus, fed diets containing levels of pyridoxine and haematological response under heat stress

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)