Effects of protected sodium butyrate (PSB) in broiler diets with reduced metabolizable energy (ME) content on production performance, intestinal histomorphometry and nutrient metabolizability were evaluated in two experiments. In one experiment, 384 one-day-old male chicks with an average weight of 44 ± 0.2 g were distributed into four treatment groups in a completely randomized design with six replications of 16 birds. The treatments consisted of i) a basal diet without reduced ME or PSB (BD); ii) BD with a 100 kcal/kg reduction in ME, without PSB; iii) BD with a 100 kcal/kg reduction in ME + 105 g/t PSB; and iv) BD with a 100 kcal/kg reduction in ME + 225 g/t PSB. Performance and carcass yield were evaluated. In the second experiment, 280 one-day-old male chicks were also distributed into four treatment groups in a completely randomized design with seven replications of 10 birds to evaluate metabolizability coefficients and intestinal development. The birds that received diet 3 had the best feed conversion ratio (FCR), final weight (FW) and weight gain (WG). Inclusion of PSB (105 and 225 g/t) in reduced-ME diets increased the metabolizability of crude protein coefficient at 35 days old. Birds that received diet 3 also had greater jejunal villus height at 21 and 35 days old. It is recommended to feed 105 g/t of PSB with a 100 kcal/kg reduction in ME until broilers were 35 days old.

Two experiments were conducted to evaluate the effect of including protected sodium butyrate (PSB) in the diets of laying hens of advanced age on productive performance and egg quality. In experiment 1, a total of 320 Dekalb White 61-week-old laying hens were distributed in a randomized block design to four treatments (0, 105, 210, and 300 g/ton of PSB) with eight replicates of 10 birds. The experiment was conducted over 112 days, divided into four periods of 28 days each. Productive performance and egg quality were evaluated. In experiment 2, a total of 58 000 Dekalb White 70-week-old laying hens were assigned to diets containing either 0 or 105 g/ton of PSB with two replicates of 14 500 birds. The experiment was conducted on a commercial poultry farm. Egg production and the percentages of broken eggs and dirty eggs were evaluated. The PSB levels did not influence the productive performance of laying hens. A quadratic effect was observed for eggshell thickness, percentage of eggshell and eggshell strength, reaching maxima at 193, 136 and 198 g/ton of PSB, respectively. A quadratic effect on yolk index reached a minimum at 181 g/ton of PSB. The percentages of dirty eggs and broken eggs were lower in laying hens fed 105 g/ton of PSB. In conclusion, the addition of PSB to the diets of old laying hens improved the eggshell quality and decreased the percentage of broken and dirty eggs.

The addition of protease to broiler diets may complement the action of endogenous enzymes and improve protein digestibility. Here, the authors evaluated the effect of adding protease to broiler diets that contained animal-based meal on bird performance, digestibility, and biometry of digestive tract organs. Four treatments, which contained animal- or plant-based meals with or without supplementary protease were compared, namely basal vegetable feed (BVF), BVF + protease (BFP), basal vegetable feed + animal byproduct meal (BFA), and BFA + protease. In the first experiment, 320 one-day-old Cobb 500® chicks were allocated to eight replicates with 10 birds per replicate. The experimental period was seven days, and nutrient metabolizability was evaluated. In the second experiment, 720 one-day-old Cobb 500® chicks were assigned to treatments in a similar manner, with six replicates and 30 birds per replicate, but the experimental period was 42 days. Significant differences (P <0.05) were observed between treatments for nutrient digestibility, weight gain, feed consumption, average final weight, food conversion, viability, and biometry of the pancreas. Treatments with animal-based meals had the highest digestibility. Birds fed these meals grew faster in the pre-starter phase and consumed less feed between 1 and 21 days. However, between 1 and 42 days old, broilers fed plant-based diets had better feed conversion, and the group that did not receive protease supplementation had a better liveability rate.

J. M. G. Carvalho, E.M. Bonfim-Silva, T.J.A. Silva, Helon H.F. Sousa, S.L. Guimarães, and A.B. Pacheco. 2016. Nitrogen and potassium in production, nutrition and water use efficiency in wheat plants. Cien. Inv. Agr. 43(3):442-451. Wheat (Triticum aestivum L.) is a cereal used in food and feed for its nutritional properties. For nitrogen and potassium management, efficient use of water is required for greater availability and transportation of these nutrients. The objective of this study was to evaluate the effect of combinations of nitrogen and potassium doses in production, nutrition and water use efficiency in wheat. The experiment was held in a greenhouse with 8-dm³ pots using Oxisol collected at a depth of 0-0.2 m under Cerrado vegetation. Base saturation was raised to 60%. The experimental design was a randomized block in a 5x5 factorial arrangement with five doses of nitrogen (0, 100, 200, 300 and 400 mg dm-3) and potassium (0, 90, 180, 270, 360 mg dm-3) with four replications. Soil moisture was maintained by an auto-irrigation system (3 kPa pressure). The experiment was collected at 95 days after plant emergence. We evaluated the dry mass of ears, shoots and roots, the total dry matter, nitrogen and potassium in grains, and the consumption and efficiency in water use. An analysis of variance and regression test at 5% probability was conducted using SAS statistical software (SAS Institute, Inc., Cary, North Carolina). The nitrogen and potassium alone increased the dry mass of ears by 31.47% and 20.91%, respectively. There was an interaction of nutrients on the dry mass of the shoots and the total dry matter. Potassium increases the concentration of nitrogen. Nitrogen and potassium promote gains in production, nutrition and water use efficiency.

. M. G. Carvalho, E.M. Bonfim-Silva, T.J.A. Silva, H.H.F. Sousa, S.L. Guimarães y A.B. Pacheco. 2016. Nitrógeno y potasio en la producción, nutrición y eficiencia del uso de agua en plantas de trigo. Cien. Inv. Agr. 43(3):442-451. Trigo (Triticum aestivum L.) es un cereal utilizado en la alimentación humana y animal por sus propiedades nutritivas. En la gestión de nitrógeno y potasio, el uso eficiente del agua es necesaria para la mayor disponibilidad y el transporte de estos nutrientes. Este estudio tuvo como objetivo evaluar el efecto de las combinaciones de nitrógeno y potasio en la producción, la nutrición y la eficiencia en el uso del agua en trigo. El experimento se realizó en un invernadero con recipientes de 8 dm³. Usando Oxisol, recogido a una profundidad de 0-0,2 m de área bajo la vegetación del Cerrado. La saturación de bases se aumentó a 60%. El diseño experimental fue de bloques al azar en arreglo factorial 5x5 con cinco dosis de nitrógeno (0, 100, 200, 300 y 400 mg dm-3) y potasio (0, 90, 180, 270 y 360 mg dm-3) con cuatro repeticiones. La humedad del suelo se mantuvo con un sistema autoirrigante (mantenido en el potencial de -3 kPa). El experimento foe cosechó después de 95 días de la emergencia. Se evaluaron la materia seca de las espigas, brotes, raíces, y materia seca vegetal total; nitrógeno y potasio en granos; el consumo y la eficiencia en el uso del agua. Las variables se sometieron a análisis de varianza y prueba de regresión al 5% de probabilidad por el programa estadístico SAS. El nitrógeno y potasio, por separado, aumentaron el peso seco de las espigas, con incrementos de 31,47 y 20,91%, respectivamente. Hubo interacción de nutrientes a la masa seca de la parte aérea y total. El potasio aumenta la concentración de nitrógeno. El nitrógeno y el potasio promueven ganancias en la producción, la nutrición y la eficiencia en el uso del agua.

A abrangência dos estudos sobre a toxemia da prenhez (TP) tem sido constante e inovadora em alguns conceitos, porém o estudo dos casos clínicos naturais em cabras é inexistente no Brasil. Diante disso, realizou-se análise das alterações clínicas, do perfil bioquímico, eletrolítico e hormonal com o objetivo de compreender a fisiopatogenia da enfermidade. Foram avaliadas 22 cabras atendidas na Clínica de Bovinos, Campus Garanhuns/UFRPE, diagnosticadas com TP, no período de 2007 a 2012. Realizou-se exame clínico e coleta de sangue e urina, para análise do perfil bioquímico e hormonal. Acentuadas alterações clínicas foram observadas, como decúbito, apatia, dispneia, aumento da temperatura corpórea, mucosas congestas, vasos episclerais injetados, desidratação, anorexia, hipomotilidade ou atonia ruminal, edema nos membros. Os resultados laboratoriais revelaram valores elevados para ureia, creatinina, ácidos graxos não esterificados (AGNE), β-hidroxibutirato, amilase e cortisol, enquanto as variáveis: proteína total, albumina, cálcio total, cálcio ionizado, fósforo, a relação cálcio e fósforo, sódio e insulina encontraram-se diminuídos. Outros parâmetros bioquímicos e eletrolíticos, como frutosamina, globulina, cloro, potássio e magnésio mantiveram-se dentro da faixa de normalidade. Quanto a glicemia, foram constatados três condições nas cabras com TP, normoglicêmicas (50%), hiperglicêmicas (27,28%) e hipoglicêmicas (22,72%). Dos 17 (77,27%) animais que obtiveram alta 10 (58,82%) eram normoglicêmicos. Vieram a óbito 22,8 % (5/22), no qual 60% (3/5) delas estavam hiperglicêmicas. A média de nascimento por cabra foi superior a duas crias. Verificou-se existir associação fortemente positiva dos AGNE's com a albumina (r=0,60), fraca com o β-hidroxibutirato (r=0,10) e glicose (r=0,03). Porém, existiu relação fortemente negativa dos AGNE's com a insulina (r= -0,70), moderada com o cortisol (r= -0,52) e amilase (r= -0,30). Marcadas alterações clínicas e metabólicas são observadas em cabras com TP, em que o aumento de certos metabólitos do perfil energético e hormonal se torna importante ferramenta para o diagnóstico e constitui um bom instrumento para avaliar a magnitude da condição clínica dos animais enfermos. Os dados encontrados servem de referência para estudos sobre nutrição e metabolismo em cabras com TP.

Studies on pregnancy toxemia (PT) have been constant and innovative with regard to some concepts. However, no studies have been carried out on natural clinical cases in Brazil. Thus, the aim of the present study was to perform an analysis of clinical, biochemical, electrolytic and hormonal findings in goats with PT to gain an understanding of the physiopathology of this metabolic disease. Twenty-two goats with a diagnosis of PT were analyzed at the Bovine Clinic of the Federal Rural University of Pernambuco (Garanhuns Campus, Brazil) between 2007 and 2012. Clinical exams were performed and both blood and urine were collected for the biochemical and hormonal analyses. Accentuated clinical changes were found, such as decubitus, apathy, dyspnea, increased body temperature, congested mucous membranes, dilated episcleral vessels, dehydration, anorexia, ruminal stasis and edema in the limbs. The laboratory findings revealed high concentrations of urea, creatinine, non-esterified fatty acids, β-hydroxybutyrate, amylase and cortisol as well as reductions in total proteins, albumin, total calcium, ionized calcium, phosphorus the calcium to phosphorus ratio, sodium and insulin. Fructosamine, globulin, chlorine, potassium and magnesium were within the range of normality. Regarding blood sugar, 50% of the goats were within the normal range, 27.28% were hyperglycemic and 22.72% were hypoglycemic; among the 17 animals (77.27%) discharged with life, ten (58.82%) had normal blood sugar levels. Five goats (22.8%) died, 60% (3/5) of which were hyperglycemic. The mean birth rate per goat was more than two offspring. Non-esterified fatty acids were strongly associated with albumin (r = 0.60) and weakly associated with both β-hydroxybutyrate (r=0.10) and glucose (r=0.03). Moreover, non-esterified fatty acids were negatively associated with insulin (r= -0.70; strong association), cortisol (r= -0.52; moderate association) and amylase (r= -0.30; moderate association). Striking clinical and metabolic alterations are found in goats with PT. The increase in certain metabolites of the energy and hormonal profile constitutes an important tool for the diagnosis and determination of the magnitude of the clinical condition in affected animals. The present data can serve as reference for future studies on nutrition and metabolism in goats with pregnancy toxemia.

This study aimed to test different protocols for the extraction of microbial DNA from the coral Mussismilia harttii. Four different commercial kits were tested, three of them based on methods for DNA extraction from soil (FastDNA SPIN Kit for soil, MP Bio, PowerSoil DNA Isolation Kit, MoBio, and ZR Soil Microbe DNA Kit, Zymo Research) and one kit for DNA extraction from plants (UltraClean Plant DNA Isolation Kit, MoBio). Five polyps of the same colony of M. harttii were macerated and aliquots were submitted to DNA extraction by the different kits. After extraction, the DNA was quantified and PCR-DGGE was used to study the molecular fingerprint of Bacteria and Eukarya. Among the four kits tested, the ZR Soil Microbe DNA Kit was the most efficient with respect to the amount of DNA extracted, yielding about three times more DNA than the other kits. Also, we observed a higher number and intensities of DGGE bands for both Bacteria and Eukarya with the same kit. Considering these results, we suggested that the ZR Soil Microbe DNA Kit is the best adapted for the study of the microbial communities of corals.

The aim of this study was to determine if bone marrow mononuclear cell (BMMC) transplantation is safe for moderate to severe idiopathic dilated cardiomyopathy (IDC). Clinical trials have shown that this procedure is safe and effective for ischemic patients, but little information is available regarding non-ischemic patients. Twenty-four patients with IDC, optimized therapy, age 46 ± 11.6 years, 17 males, NYHA classes II-IV, and left ventricular ejection fraction <35% were enrolled in the study. Clinical evaluation at baseline and 6 months after stem cell therapy to assess heart function included echocardiogram, magnetic resonance imaging, cardiopulmonary test, Minnesota Quality of Life Questionnaire, and NYHA classification. After cell transplantation 1 patient showed a transient increase in enzyme levels and 2 patients presented arrhythmias that were reversed within 72 h. Four patients died during follow-up, between 6 and 12 weeks after therapy. Clinical evaluation showed improvement in most patients as reflected by statistically significant decreases in Minnesota Quality of Life Questionnaire (63 ± 17.9 baseline vs 28.8 ± 16.75 at 6 months) and in class III-IV NYHA patients (18/24 baseline vs 2/20 at 6 months). Cardiopulmonary exercise tests demonstrated increased peak oxygen consumption (12.2 ± 2.4 at baseline vs 15.8 ± 7.1 mL·kg-1·min-1 at 6 months) and walked distance (377.2 ± 85.4 vs 444.1 ± 77.9 m at 6 months) in the 6-min walk test, which was not accompanied by increased left ventricular ejection fraction. Our findings indicate that BMMC therapy in IDC patients with severe ventricular dysfunction is feasible and that larger, randomized and placebo-controlled trials are warranted.

The effectiveness of the caspase-9-based artificial "death switch" as a safety measure for gene therapy based on the erythropoietin (Epo) hormone was tested in vitro and in vivo using the chemical inducer of dimerization, AP20187. Plasmids encoding the dimeric murine Epo, the tetracycline-controlled transactivator and inducible caspase 9 (ptet-mEpoD, ptet-tTAk and pSH1/Sn-E-Fv’-Fvls-casp9-E, respectively) were used in this study. AP20187 induced apoptosis of iCasp9-modified C2C12 myoblasts. In vivo, two groups of male C57BI/6 mice, 8-12 weeks old, were injected intramuscularly with 5 µg/50 g ptet-mEpoD and 0.5 µg/50 g ptet-tTAk. There were 20 animals in group 1 and 36 animals in group 2. Animals from group 2 were also injected with the 6 µg/50 g iCasp9 plasmid. Seventy percent of the animals showed an increase in hematocrit of more than 65% for more than 15 weeks. AP20187 administration significantly reduced hematocrit and plasma Epo levels in 30% of the animals belonging to group 2. TUNEL-positive cells were detected in the muscle of at least 50% of the animals treated with AP20187. Doxycycline administration was efficient in controlling Epo secretion in both groups. We conclude that inducible caspase 9 did not interfere with gene transfer, gene expression or tetracycline control and may be used as a safety mechanism for gene therapy. However, more studies are necessary to improve the efficacy of this technique, for example, the use of lentivirus vector.

Os valores hematológicos e bioquímicos, incluindo os eletrólitos plasmáticos e os hormônios da tireoide, foram determinados em 73 ovelhas, clinicamente saudáveis, da raça Churra da Terra Quente, raça ovina característica do nordeste de Portugal. Os valores obtidos para o hemograma foram: eritrócitos 9,8±1,5×10(12) /L; hemoglobina 118,1±19,1g/L; hematócrito 40,8±5,9%; leucócitos 5,7±1,8×10(9) /L e plaquetas 544,3±177,2×10(9)/L. O tempo de trombina foi de 17,3±1,7 segundos. Os valores dos parâmetros bioquímicos avaliados foram: proteínas totais 76,4±6,1g/L; glicose 2,87±0,60mmol/L; colesterol total 1,65±0,33mmol/L; aspartato amino transferase 155,9±49,2U/L; alanina amino transferase 23,2±9,6U/L; gama-glutamil transferase 48,0±18,7U/L; fosfatase alcalina total 121,6±76,1U/L; glutamato desidrogenase 6,4±3,7U/L; ureia 7,32±2,22mmol/L; creatinina 123,0±54,1μmol/L; cálcio total 2,53±0,25mmol/L; fósforo 2,10±0,46mmol/L e magnésio 1,01±0,09mmol/L; sódio 152,04±3,65mmol/L; potássio 4,7±0,4mmol/L e cálcio ionizado 1,32±0,07mmol/L; tiroxina total 111,75±42,29nmol/L; tri-iodotironina total 1,01±0,28nmol/L; T4 livre 11,93±1,78pmol/L; T3 livre 4,22±1,33pmol/L e hormônio estimulante da tireoide 0,18±0,19μIU/mL. Apesar de terem sido observadas algumas diferenças entre a raça Churra da Terra Quente e outras raças, os valores hematológicos e bioquímicos, eletrólitos plasmáticos e hormônios da tireóide, desta raça autóctone apresentam-se no geral situados dentro dos intervalos de referência publicados para a espécie ovina.

Hematological and biochemical parameters, including plasma electrolytes and thyroid hormones, were determined in 73 clinically healthy Churra-da-Terra-Quente ewes, a typical breed from the northeast of Portugal. The hemogram values were: erythrocytes 9.8±1.5×10(12)/L; haemoglobin 118.1±19.1g/L; haematocrit 40.8±5.9%; leukocytes 5.7±1.8×10(9)/L; and platelets 544.3±177.2×10(9)/L. The thrombin time was 17.3±1.7 seconds. The values of biochemical parameters were: total protein 76.4±6.1g/L; glucose 2.87±0.60mmol/L; total cholesterol 1.65±0.33mmol/L; aspartate aminotransferase 155.9±49.2U/L; alanine aminotransferase 23.2±9.6U/L; γ-glutamyl transferase 48.0±18.7U/L; total alkaline phosphatase 121.6±76.1U/L; glutamate dehydrogenase 6.4±3.7U/L; urea 7.32±2.22mmol/L; creatinine 123.0±54.1μmol/L; total calcium 2.53±0.25mmol/L; phosphorus 2.10±0.46mmol/L; magnesium 1.01±0.09mmol/L; sodium 152.04±3.65mmol/L; potassium 4.7±0.4mmol/L; ionized calcium 1.32±0.07mmol/L; total thyroxine 111.75±42.29nmol/L; total triiodothyronine 1.01±0.28nmol/L; free T4 11.93±1.78pmol/L; free T3 4.22±1.33pmol/L; and thyroid-stimulating hormone 0.18±0.19μIU/mL. Although differences among the Churra-da-Terra-Quente breed and other breeds may occur, the hematological and biochemical parameters, plasma electrolytes, and thyroid hormones, for this indigenous breed, were generally situated within the reference intervals previously reported for sheep.

This review focuses on the mechanisms of DNA methylation, DNA methylation pattern formation and their involvement in gene regulation. Association of DNA methylation with imprinting, embryonic development and human diseases is discussed. Furthermore, besides considering changes in DNA methylation as mechanisms of disease, the role of epigenetics in general and DNA methylation in particular in transgenerational carcinogenesis, in memory formation and behavior establishment are brought about as mechanisms based on the cellular memory of gene expression patterns.

Relatamos o caso de uma gestante de 26 anos apresentando episódios de sangramento vaginal, perda ponderal e dispnéia, que recebeu o diagnóstico de coriocarcinoma com metástases pulmonares. O nível de gonadotrofina coriônica (hCG) era >2,5 x 10(6)mU/mL. O TSH era de 0,037mU/L (0,49 - 4,67), o T4 de 18,1ug/dL (4,9 - 10,7), e o T3 de 136ng/dL (45 - 137), confirmando o quadro de hipertireoidismo subclínico induzido pela hCG. A paciente foi submetida a um regime combinado de quimioterapia com etoposídeo, metotrexate e dactiomicina. A evolução inicial foi complicada por um quadro de urosepsis com insuficiência respiratória, necessitando entubação endotraqueal por 3 dias. Após, houve melhora progressiva com normalização dos testes de função tireoideana no 12º dia de internamento. Após 6 meses e 10 ciclos de quimioterapia, a paciente estava em remissão e sem sinais de tumor residual ou de atividade paraneoplásica dependente de hCG.

A 26-year-old pregnant woman presenting with repeated episodes of vaginal bleeding, weight loss, and shortness of breath was diagnosed with choriocarcinoma with metastases to both lungs. Chorionic gonadotropin levels (hCG) were >2.5 x 10(6)mU/mL. Consistent with hCG-induced subclinical hyperthyroidism, she had a suppressed TSH of 0.037mU/L (0.49 - 4.67), a T4 of 18.1µg/dL (4.9 - 10.7), and T3 of 136ng/dL (45 - 137). Chemotherapy with a combined regimen with etoposide, methotrexate, and dactinomycine was started. The initial course was complicated by urosepsis with respiratory distress requiring endotracheal intubation for 3 days. She then improved rapidly, and her thyroid function tests were within normal limits by day 12. Six months later, after ten cycles of chemotherapy, the patient was in remission without signs of residual tumor or hCG-induced paraneoplastic activity.

Complex interactions between androgen and estrogen (E2) regulate prostatic development and physiology. We analyzed the early effects of a high single dose of E2 (25 mg/kg body weight) and castration (separately or combined) on the adult 90-day-old male Wistar rat ventral prostate. Androgen levels, prostate weight, and the variation in the relative and absolute volume of tissue compartments and apoptotic indices were determined for 7 days. Castration and exogenous E2 markedly reduced ventral prostate weight (about 50% of the control), with a significant reduction in the epithelial compartment and increased stroma. The final volume of the epithelium was identical at day 7 for all treatments (58.5% of the control). However, E2 had an immediate effect, causing a reduction in epithelial volume as early as day 1. An increase in smooth muscle cell volume resulted from the concentration of these cells around the regressing epithelium. The treatments resulted in differential kinetics in epithelial cell apoptosis. Castration led to a peak in apoptosis at day 3, with 5% of the epithelial cells presenting signs of apoptosis, whereas E2 caused an immediate increase (observed on day 1) and a sustained (up to day 7) effect. E2 administration to castrated rats significantly increased the level of apoptosis by day 3, reaching 9% of the epithelial cells. The divergent kinetics between treatments resulted in the same levels of epithelial regression after 7 days (~30% of control). These results show that E2 has an immediate and possibly direct effect on the prostate, and anticipates epithelial cell death before reducing testosterone to levels as low as those of castrated rats. In addition, E2 and androgen deprivation apparently cause epithelial cell death by distinct and independent pathways.

The structure of the large proteoglycan present in the bullfrog epiphyseal cartilage was studied by immunochemical and biochemical methods. The isolated monomer showed a polydisperse behavior on Sepharose CL2B, with a peak at Kav = 0.14. Chondroitin sulfate chains were identified by HPLC analysis of the products formed by chondroitinase digestion and mercuric acetate treatment. These chains have approximately 38 disaccharides, a Di45:Di68 ratio of 1.6 and GalNAc4S + GalNAc4,6S are the main non-reducing terminals. Keratan sulfate was identified by the use of two monoclonal antibodies in Western blots after chondroitinase ABC treatment. A keratan sulfate-rich region (~110 kDa) was isolated by sequential treatment with chondroitinase ABC and proteases. We also employed antibodies in Western blotting experiments and showed that the full length deglycosylated core protein is about 300 kDa after SDS-PAGE. Domain-specific antibodies revealed the presence of immunoreactive sites corresponding to G1/G2 and G3 globular domains and the characterization of this large proteoglycan as aggrecan. The results indicate the high conservation of the aggrecan domain structure in this lower vertebrate.