Valores hematológicos y bioquímicos, y su asociación con el estado nutricional, en escolares urbanos. Cuenca

Objetivo: Determinar los valores hematológicos y bioquímicos y su asociación con el estado nutricional, en escolares urbanos de Cuenca. Materiales y métodos: Estudio transversal analítico, muestra aleatorizada por conglomerados, de 585 escolares entre 5 y 12 años de las escuelas urbanas de Cuenca, periodo 2012-2013. Se determinó el estado nutricional usando curvas de crecimiento para niños yadolescentes (z-score), índice de masa corporal por edad y sexo, según las recomendaciones de la Organización Mundial de la Salud; con los valores de los análisis hematológicos y bioquímicos practicados, se relacionaron los resultados de laboratorio con el estado nutricional. Resultados: La media de edad en la población fue de 8.87±1.9 años; promedio en valores hematológicos: glóbulos rojos 4.82±0.31 x 106/ml; glóbulos blancos 7.15±1.86 x 103/ ml; hemoglobina 13.66±1.34 g/dL; hematocrito 40.59±3.92%; volumen corpuscular medio (VCM) 83.64±4.03fl; hemoglobina corpuscular media (HCM) 38.31±1.27pg. Promedio de valores bioquímicos: proteína total 6.92±1.04g/ dL, albúmina sérica 4.37±0.7g/dL, hierro sérico 87.40±35.31μg/dL. El 19.3% presentó malnutrición (desnutrición 1.2%, sobrepeso 12.8% y obesidad 5.3%), existiendo asociación estadísticamente significativa (p<0.05) con glóbulos rojos. Existieron diferencias estadísticamente significativas, entre niños y niñas, en valores de hierro, VCM y HCM; y respecto de malnutrición, en hierro sérico, glóbulos rojos, glóbulos blancos, hematocrito y VCM. Conclusiones: Se encontró asociación entre el estado nutricional y los valores hematológicos y bioquímicos. Se determinó prevalencia alta de sobrepeso y obesidad en población escolar urbana de Cuenca, similar a la reportada en otros estudios.

Acute lethal and sublethal effects of neem leaf extract on the neotropical freshwater fish Prochilodus lineatus

The aim of this study was to determine the toxicity of the aqueous extract of neem leaves, a product extensively used in fish-farms as alternative for the control of fish parasites and fish fry predators, for the neotropical fish Prochilodus lineatus. The 24 It LC(50) of neem leaf extract for juveniles P lineatus was estimated as 4.8 g L(-1); the fish were then exposed for 24 h to 2.5, 5.0 and 7.5 g L(-1) or only clean water (control). Plasma glucose levels were higher in fish exposed to 2.5 g L(-1) and 5.0 g L(-1) neem extract, relative to control, indicating a typical stress response. Neem extract did not interfere with the osmoregulating capacity of the fish, as their plasma sodium, chloride, total protein and osmolarity did not change. The presence of the biopesticide interfered with the antioxidant defense system of P. lineatus, as there was a decrease in liver catalase activity at all neem concentrations and the detoxifying enzyme glutathione-S-transferase was activated in fish exposed to 5.0 g L(-1). Fish exposed to all neem extract concentrations exhibited damaged gill and kidney tissue. These results indicate that although neem extract is less toxic to P. lineatus than other synthetic insecticides used in fish-farming it does cause functional and morphological changes in this fish species. (c) 2006 Elsevier B.V. All rights reserved.

Sodium chloride added to transport water and physiological responses of Matrinxã Brycon amazonicus (Teleost: Characidae)

A adição de sal à água tem sido utilizada para a mitigação de estresse e aumento da taxa de sobrevivência em peixes. O presente estudo avaliou o efeito do cloreto de sódio (0,0; 1,0; 3,0 e 6.0 g/l) nas concentrações de cortisol plasmático, glicemia, triglicerídios, proteínas total plasmática, hematócrito, hemoglobina, número de eritrócitos, glicogênio e lipídio hepáticos, e lipídio muscular em matrinxã Brycon amazonicum adultos após quatro horas de transporte e durante período de recuperação de 96 h. Amostras foram coletadas antes e depois do transporte, bem como 24 e 96 h após a chegada. O nível de cortisol plasmático estava mais elevado logo após o transporte quando comparado à condição inicial (pré-transporte), exceto para os peixes transportados com sal nas concentrações 3,0 e 6,0 g/l. Comportamento semelhante foi observado para a glicemia, porém os peixes dos tratamentos 0,0, 1,0 e 3,0 g/l necessitaram de período superior a 24 h para recuperar a condição inicial. Foram registrados níveis mais baixos de glicogênio hepático em peixes do tratamento controle (0,0 g/l). Os parâmetros hemoglobina, número de eritrócitos, proteína plasmática total e lipídio hepático não apresentaram alterações durante o período experimental. Os valores de hematócrito diminuíram logo após o transporte em todos os tratamentos, retornando aos níveis iniciais após 24 h. Todos os tratamentos apresentaram redução nos níveis de lipídio muscular e triglicerídios durante o período de recuperação. Os resultados sugerem que a adição de 6,0 g/l de sal na água de transporte reduz as alterações fisiológicas de estresse e que é necessário período de 96 h após o transporte para a recuperação da condição inicial de matrinxãs transportados sem a adição de sal.

Incorporation des protéines de canola dans du pain sans gluten : impact technologique et modélisation du processus de cuisson

L’objectif de cette maîtrise est de développer une matrice alimentaire sans gluten par l’incorporation des protéines de canola (PC) dans une farine de riz blanc afin d’offrir aux personnes intolérantes au gluten un produit de bonne qualité d’un point de vue organoleptique (volume massique, structure alvéolaire et couleur) et de valeur nutritionnelle. La matrice sélectionnée est du pain à base de farine de riz blanc. Cinq formulations ont été testées dans la première partie de ce travail : témoin-1 (blé tendre), témoin-2 (100% riz), pain de riz +3% PC, pain de riz + 6% PC, pain de riz + 9% PC. Les produits obtenus ont été caractérisés à toutes les étapes de fabrication en utilisant différentes techniques : poussée volumique, variation thermique au cours des étuvages et de la cuisson, pH (acidité), perte d’eau, volume massique, analyse colorimétrique, dosage des protéines et analyse du profil de la texture. Dans la deuxième partie, deux variables indépendantes ont été additionnées; soit shortening (1, 2, 3%) et gomme de xanthane (0.5, 1, 1.5%), dans le but d’améliorer le volume massique de la meilleure formulation obtenue dans l’étape précédente. Ensuite, des essais de correction ont été attribués aux produits obtenus par l’introduction du bicarbonate de sodium (0.5, 1, 1.5%) et d’huile de canola à la place du shortening (1, 2, 3%). Les tests de panification ont donné différents volumes massiques et structures alvéolaires qui étaient jugés de qualité inférieure à celle du témoin-1 (2.518 mL/g), mais largement supérieure à celle du témoin-2 (1.417 mL/g). Le meilleur volume massique obtenu est de 1.777 mL/g, correspondant à celui du pain obtenu par la combinaison 6%PC+0.5%GH+B 1.5%+ H3%. Finalement, les résultats de ce projet ont montré l’impact positif de l’incorporation des protéines de canola dans un pain sans gluten à base de farine de riz blanc. Ce travail constitue une contribution à la possibilité de substituer le gluten par d’autres protéines ayant de bonnes propriétés techno-fonctionnelles, en particulier la capacité à donner du volume au produit final.

Diferenciación molecular de Lemna aequinoctialis y L. Valdiviana (lemnaceae) por patrones proteicos

El estudio se llevó a cabo en el Laboratorio de Genética Molecular de la Universidad Nacional, Heredia, Costa Rica. Se utilizaron plantas acuáticas macrófitas de Lemna valdiviana y L. aequinoctialis previamente identificadas. Estas especies habitan en aguas ricas en sales disueltas y aguas servidas. Recientemente se les consideró como un indicador de la contaminación ocasionada por pesticidas y herbicidas en el agua. La composición proteica llega a niveles entre 38 y 43% de materia seca, importante para diversos usos. La extracción de proteínas totales, por medio de electroforesis discontinua en gel de poliacrilamida, se realizó en condiciones reductoras con SDS (SDS-PAGE). Los geles separadores empleados fueron al 8 y 10%, gel espaciador al 4% y la tinción se efectuó con azul commassie. Los diversos patrones proteicos reflejaron diferencias genéticas entre ambas especies que se pueden deber a las diferencias de hábitat y de reproducción.

A membrane process to extract Citrus Lemon derived extracellular vesicles

Extra cellular vesicles are membrane bound and lipid based nano particles having the size range of 30 to 1000 nm released by a plethora of cells. Their prime function is cellular communication but in the recent studies, the potential of these vesicles to maintain physiological and pathological processes as well as their nano-sized constituents opened doors to its applications in therapeutics, and diagnostics of variety of diseases such as cancer. Their main constituents include lipids, proteins, and RNAs. They are categorized into subtypes such as exosomes, micro-vesicles and apoptotic bodies In recent studies, extracellular vesicles that are derived from plants are gaining high regard due to their variety of advantages such as safety, non-toxicity, and high availability which promotes large scale production. EVs are isolated from mammalian and plant cells using multitude of techniques such as Ultracentrifugation, SEC, Precipitation and so on. Due to the variety in the sources as well as shortcomings arising from the isolation method, a scalable and inexpensive EV isolation method is yet to be designed. This study focusses on isolation of EVs from citrus lemon juice through diafiltration. Lemon is a promising source due to its biological properties to act as antioxidant, anticancer, and anti-inflammatory agents. Lemon derived vesicles was proven to have several proteins analogous to mammalian vesicles. A diafiltration could be carried out for successful removal of impurities and it is a scalable, continuous technique with potentially lower process times. The concentration of purified product and impurities are analysed using Size Exclusion Chromatography in analytical mode. It is also considered imperative to compare the results from diafiltration with gold standard UC. BCA is proposed to evaluate total protein content and DLS for size measurements. Finally, the ideal mode of storage of EVs to protect its internals and its structure is analysed with storage tests.

Inhibition of phospholipase A(2) in rat brain decreases the levels of total Tau protein

The microtubule-associated protein Tau promotes the assembly and stability of microtubules in neuronal cells. Six Tau isoforms are expressed in adult human brain. All six isoforms become abnormally hyperphosphorylated and form neurofibrillary tangles in Alzheimer disease (AD) brains. In AD, reduced activity of phospholipase A(2) (PLA(2)), specifically of calcium-dependent cytosolic PLA(2) (cPLA(2)) and calcium-independent intracellular PLA(2) (iPLA(2)), was reported in the cerebral cortex and hippocampus, which positively correlated with the density of neurofibrillary tangles. We previously demonstrated that treatment of cultured neurons with a dual cPLA(2) and iPLA(2) inhibitor, methyl arachidonyl fluorophosphonate (MAFP), decreased total Tau levels and increased Tau phosphorylation at Ser(214) site. The aim of this study was to conduct a preliminary investigation into the effects of in vivo infusion of MAFP into rat brain on PLA(2) activity and total Tau levels in the postmortem frontal cortex and dorsal hippocampus. PLA(2) activity was measured by radioenzymatic assay and Tau levels were determined by Western blotting using the anti-Tau 6 isoforms antibody. MAFP significantly inhibited PLA(2) activity in the frontal cortex and hippocampus. The reactivity to the antibody revealed three Tau protein bands with apparent molecular weight of close to 40, 43 and 46 kDa in both brain areas. MAFP decreased the 46 kDa band intensity in the frontal cortex, and the 43 and 46 kDa band intensities in the hippocampus. The results indicate that in vivo PLA(2) inhibition in rat brain decreases the levels of total (nonphosphorylated plus phosphorylated) Tau protein and corroborate our previous in vitro findings.

A cassette ligation strategy with thioether replacement of three Gly-Gly peptide bonds: Total chemical synthesis of the 101 residue protein early pregnancy factor [psi(CH2S)(28-29,56-57,76-77)]

The 101 residue protein early pregnancy factor (EPF), also known as human chaperonin 10, was synthesized from four functionalized, but unprotected, peptide segments by a sequential thioether ligation strategy. The approach exploits the differential reactivity of a peptide-NHCH2CH2SH thiolate with XCH2CO-peptides, where X = Cl or I/Br. Initial model studies with short functionalized (but unprotected) peptides showed a significantly faster reaction of a peptide-NHCH2CH2SH thiolate with a BrCH2CO-peptide than with a CICH2CO-peptide, where thiolate displacement of the halide leads to chemoselective formation of a thioether surrogate for the Gly-Gly peptide bond. This rate difference was used as the basis of a novel sequential ligation approach to the synthesis of large polypeptide chains. Thus, ligation of a model bifunctional N-alpha-chloroacetyl, C-terminal thiolated peptide with a second N-alpha-bromoacetyl peptide demonstrated chemoselective bromide displacement by the thiol group. Further investigations showed that the relatively unreactive N-alpha-chloroacetyl peptides could be activated by halide exchange using saturated KI solutions to yield the highly reactive No-iodoacetyl peptides. These findings were used to formulate a sequential thioether ligation strategy for the synthesis of EPF, a 101 amino acid protein containing three Gly-Gly sites approximately equidistantly spaced within the peptide chain. Four peptide segments or cassettes comprising the EPF protein sequence (BrAc-[EPF 78-101] 12, ClAc-[EPF 58-75]-[NHCH2CH2SH] 13, ClAc-[EPF 30-55]-[NHCH2CH2SH] 14, and Ac-[EPF 1-27]-[NHCH2CH2SH] 15) of EPF were synthesized in high yield and purity using Boc SPPS chemistry. In the stepwise sequential ligation strategy, reaction of peptides 12 and 13 was followed by conversion of the N-terminal chloroacetyl functional group to an iodoacetyl, thus activating the product peptide for further ligation with peptide 14. The process of ligation followed by iodoacetyl activation was repeated to yield an analogue of EPF (EPF psi(CH2S)(28-29,56-57,76-77)) 19 in 19% overall yield.

Influence of ageing, heat shock treatment and in vivo total antioxidant status on gene-expression profile and protein synthesis in human peripheral lymphocytes

Ageing results in a progressive, intrinsic and generalised imbalance of the control of regulatory systems. A key manifestation of this complex biological process includes the attenuation of the universal stress response. Here we provide the first global assessment of the ageing process as it affects the heat shock response, utilising human peripheral lymphocytes and cDNA microarray analysis. The genomic approach employed in our preliminary study was supplemented with a proteomic approach. In addition, the current study correlates the in vivo total antioxidant status with the age-related differential gene expression as well as the translational kinetics of heat shock proteins (hsps). Most of the genes encoding stress response proteins on the 4224 element microarray used in this study were significantly elevated after heat shock treatment of lymphocytes obtained from both young and old individuals albeit to a greater extent in the young. Cell signaling and signal transduction genes as well as some oxidoreductases showed varied response. Results from translational kinetics of induction of major hsps, from 0 to 24 It recovery period were broadly consistent with the differential expression of HSC 70 and HSP 40 genes. Total antioxidant levels in plasma from old individuals were found to be significantly lower by comparison with young, in agreement with the widely acknowledged role of oxidant homeostasis in the ageing process. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.

Adverse effects of high dose carnitine supplementation of total parenteral nutrition on protein and fat oxidation in the critically ill.

The aim of the present study was to investigate the effects of continuous and acute L-carnitine supplementation of total parenteral nutrition (TPN) on protein and fat oxidation in severe catabolism. A critically ill and severely malnourished male patient received TPN (non protein energy = 41 kcal/kg/day, provided equally as fat and glucose) over 38 days, without L-carnitine for 23 days and with carnitine supplements (15 mg/kg/day) for the following 15 days. Subsequently, he was given carnitine-free enteral nutrition for 60 more days. A four-hour infusion of 100 mg L-carnitine was given on day 11 of each TPN period. Indirect calorimetry was carried out after 11 days of either carnitine-free or supplemented TPN and at the initiation of enteral nutrition. Additional measurements were performed 4 hours and 24 hours after the acute infusions of carnitine. The rate of protein oxidation and the respiratory quotient were found to be higher, and the rate of fat oxidation to be lower, with carnitine-supplemented TPN, than with either carnitine-free TPN or enteral nutrition. Acute infusion of carnitine resulted in an increased rate of protein oxidation and a reduced rate of fat oxidation on both TPN-regimens. These unfavourable effects on protein metabolism may be due to an impairment of fat oxidation by excess amounts of carnitine.

Clinical, hematological, total plasma protein and fibrinogen parameters of magellanic penguins ( Spheniscus magellanicus ) pre- and post-rehabilitation

Abstract: Magellanic penguins (Spheniscus magellanicus) usually arrive in poor body conditions at Brazilian beaches during the winter. Hematology provides valuable information about clinical and immunity status of the animals. The aims of this study were to determine the hematologic, total plasma protein (TPP) and fibrinogen profiles of young and adult magellanic penguins in PROAMAR and CETAS-SC, relating these results with the state of health and survival possibility of the animals. In Paraná 14 animals were evaluated in pre and eight in post-rehabilitation and 29 animals were evaluated in Santa Catarina after rehabilitation. Before rehabilitation, all animals showed weakness. In hematological exams of these animals, we found that anemia was present in 83% of the penguins that died and 50% of those which survived. The heterophils/lymphocytes (H/L) ratio was 3.87±0.57 in animals that died, significantly higher than the average of 2.20±0.30 for animals that survived. These two parameters are useful to assess the survival possibility of animals to rehabilitation. The body condition score was positively correlated with hematocrit and TPP, and negatively correlated with H/L ratio. After rehabilitation, the values were similar to other animals of the family Spheniscidae, with averages ranging from 1.64 to 1.90x106 erythrocytes/μL; 43.38 to 48.80% of hematocrit; 12.45 to 13.52g/dL of hemoglobin; 8,684 to 14,011 leukocytes/μL; 4,767 to 8,041 heterophils/μL; 3,215 to 4,951 lymphocytes/μL; 95 to 655 eosinophils/μl; 179.8 to 277.9 monocytes/μL; 141 to 184.9 basophils/μL; and 1.26 to 1.74 of H/L ratio. These parameters can therefore be used as reference values and release parameters for young and adult Magellanic penguins in captivity on the rehabilitation centers.

Experimental infection of commercial layers with wild or attenuated Salmonella Gallinarum mutant strains: Anatomic pathology, total blood cell count and serum protein levels

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

Isolated total RNA and protein are preserved after thawing for more than twenty-four hours

OBJECTIVE: The preservation of biological samples at a low temperature is important for later biochemical and/or histological analyses. However, the molecular viability of thawed samples has not been studied sufficiently in depth. The present study was undertaken to evaluate the viability of intact tissues, tissue homogenates, and isolated total RNA after defrosting for more than twenty-four hours. METHODS: The molecular viability of the thawed samples (n = 82) was assessed using the A260/A280 ratio, the RNA concentration, the RNA integrity, the level of intact mRNA determined by reverse transcriptase polymerase chain reaction, the protein level determined by Western blotting, and an examination of the histological structure. RESULTS: The integrity of the total RNA was not preserved in the thawed intact tissue, but the RNA integrity and level of mRNA were perfectly preserved in isolated defrosted samples of total RNA. Additionally, the level of beta-actin protein was preserved in both thawed intact tissue and homogenates. CONCLUSION: Isolated total RNA does not undergo degradation due to thawing for at least 24 hours, and it is recommended to isolate the total RNA as soon as possible after tissue collection. Moreover, the protein level is preserved in defrosted tissues.

Influence of total feed and protein intake on reproductive performance in the beef female through second calving /

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