Metabolismo energético, proteico e mineral de ovelhas Santa Inês hígidas e com mastite subclínica

Este estudo teve por objetivo avaliar o metabolismo energético, proteico e mineral de ovelhas Santa Inês hígidas e com mastite subclínica acompanhadas durante o final da gestação e na lactação. Foram acompanhadas ovelhas submetidas ao mesmo sistema de criação semi-intensivo. Os animais foram avaliados conforme os momentos a seguir: 10 dias que precedeu o parto (dap) e 15 dias pós parto (dpp), 30 dpp, 60 dpp e 90 dpp. Os metabólitos sanguíneos foram avaliados a partir do momento que antecedeu ao parto e os metabólitos no soro lácteo nos momentos subsequentes. Após exame clínico e bacteriológico foi realizada a triagem das ovelhas acompanhadas neste estudo, sendo 12 hígidas e 18 com mastite subclínica. Durante a lactação, mantendo os mesmos critérios de triagem, foram selecionadas 11 glândulas mamárias sadias e 20 infectadas, das quais foi colhido o leite para obtenção do soro lácteo. Foram mensurados no soro sanguíneo os metabólitos do perfil energético (ácidos graxos não esterificados (AGNEs), β-hidroxibutirato (BHB), frutosamina, colesterol e triglicérides), do perfil proteico (proteína total, albumina, uréia e creatinina) e do perfil mineral (ferro, cobre, zinco, magnésio, cálcio total, cálcio ionizado, sódio e potássio). No soro lácteo foram mensurados os íons cálcio, sódio e potássio, bem como os AGNEs e o BHB. A bioquímica sanguínea revelou haver influência (P<0,05) do período do periparto e da lactação sobre as concentrações sanguíneas dos AGNEs, BHB, colesterol, albumina, uréia, cálcio ionizado e no soro lácteo sobre o íon potássio. As ovelhas portadoras de mastite subclínica apresentaram valores sanguíneos superiores (P<0,05) de colesterol, albumina e cobre e no soro lácteo teores superiores do íon sódio e dos AGNEs e inferiores do íon potássio. O bom escore corporal das ovelhas observado durante o estudo aliado aos achados bioquímicos permitiu concluir ter ocorrido maior requerimento energético no primeiro mês da lactação, porém não o suficiente para desencadear qualquer transtorno metabólico e o aparecimento de um quadro de cetonemia, sendo estas discretas alterações mais expressivas nas ovelhas com mastite subclínica.

Perfil sanguíneo e peso de órgãos internos de bezerros leiteiros criados em diferentes sistemas de aleitamento

Objetivou-se avaliar o perfil sanguíneo e o tamanho absoluto e relativo dos órgãos internos de bezerros em sistema de aleitamento convencional ou fracionado. Foram utilizadas 10 fêmeas, com peso corporal inicial médio de 37,53 kg (±2,36) e 12 machos com peso corporal médio de 81,6 kg (±8,6), ambos da raça Holandesa. O aleitamento convencional constituiu-se de quatro litros de leite fornecidos diariamente durante 54 dias de avaliação e o fracionado, de seis litros do 6º ao 25º dias de idade, quatro litros do 26º ao 45º dias de idade e dois litros do 46º ao 59º dias de idade, além de concentrado, feno de Cynodon sp., água e suplemento mineral fornecidos ad libitum. O delineamento experimental foi inteiramente ao acaso, com cinco repetições para os parâmetros sanguíneos e seis repetições para o desenvolvimento dos órgãos internos. As coletas de sangue foram realizadas nas fêmeas nos dias sete, 21, 28, 42, 49 e 56 de idade. Para a avaliação dos órgãos internos os machos foram abatidos aos dois meses de idade, após jejum de 16 horas. Não foram observadas diferenças para os valores de proteínas totais, albumina, globulina, creatinina, ureia, colesterol e β-hidroxibutirato nas idades avaliadas (p>0,05). Entretanto, para a concentração de glicose houve interação entre os sistemas de aleitamento e os períodos avaliados, podendo ser observado que com 21 dias de idade a concentração foi maior nas bezerras mantidas em sistema fracionado, quando comparadas aos animais do grupo convencional. O sistema de aleitamento não alterou o peso corporal final, peso corporal final vazio e o peso dos órgãos internos, exceto para o coração, demonstrando que o sistema de aleitamento fracionado não altera as avaliações dos órgãos internos.

Determinação das concentrações plasmáticas de proteínas e metabólitos de cascavéis em cativeiro

Resumo: Para determinar as concentrações plasmáticas de proteínas e metabólitos de cascavéis em cativeiro, foram utilizadas 60 serpentes adultas, sendo 30 machos e 30 fêmeas. O sangue foi coletado através de punção do seio venoso paravertebral cervical e armazenado em tubos com heparina. As análises bioquímicas foram processadas colorimetricamente em Analisador Automático de Bioquímica Chemwell (Awareness Technology®, Inc). Foram calculadas as médias e desvios padrão dos seguintes constituintes: proteínas totais, albumina, globulinas, relação albumina/globulinas, ácido úrico, creatinina, ureia, colesterol, colesterol HDL e triglicérides. Os valores obtidos foram semelhantes aos descritos na literatura para repteis e serpentes, sendo as diferenças observadas provavelmente decorrentes da diferença entre espécies, clima, estação do ano e metodologia utilizada. Não houve diferenças significativas entre machos e fêmeas para os parâmetros estudados. Estes resultados podem ser úteis no estabelecimento de valores de referência para planos de conservação destes ofídios em cativeiro.

Avaliação do perfil metabólico de vacas da raça Holandesa durante o período de transição

O período de transição é um momento de grande desafio para vacas de aptidão leiteira, uma vez que, a maioria dos problemas metabólicos ocorre nesta fase podendo prejudicar toda a expectativa de produção durante a lactação, resultando em impacto econômico significativo para fazendas de produção de leite. Este trabalho teve como objetivo avaliar o perfil metabólico de vacas da raça Holandesa durante o período de transição. Doze vacas Holandesas foram avaliadas, três semanas pré-parto até três semanas pós-parto, em sistema free-stall, localizado em Inhaúma, Minas Gerais, no período de outubro a dezembro de 2012. Avaliou-se o perfil metabólico através da concentração sérica de ácidos graxos não esterificados (AGNE), beta hidroxibutirato (BHBA), colesterol (COLES), proteína total (PT), albumina (ALB), cálcio, fósforo, magnésio bem como a atividade sérica das enzimas aspartato transaminase (AST) e lactato desidrogenase (LDH). As concentrações séricas de AGNE e BHBA foram diferentes entre o pré-parto e pós-parto (p<0,05). Observou-se diminuição na concentração de COLES com a aproximação do parto com posterior aumento (p<0,05). As concentrações séricas dos minerais, PT e ALB não apresentaram diferenças (p>0,05) no período avaliado. A atividade enzimática de AST e LDH foram maiores no período pós-parto (p<0,05). A avaliação do perfil metabólico é uma importante ferramenta de monitoramento e, na situação estudada, demonstrou alterações do perfil energético das vacas entre os períodos pré e pós-parto, relacionadas provavelmente a diminuição da ingestão de alimentos. A luz dos resultados do perfil metabólico, o rebanho avaliado possui pequeno risco para a ocorrência de enfermidades no pós-parto relacionadas ao período de transição

Efeitos de fontes orgânicas de cobre e enxofre sobre os parâmetros bioquímicos no soro de ovinos

RESUMO: Minerais são componentes essenciais na dieta, exercendo diversas funções no organismo animal onde o uso de sais orgânicos visa aumentar a disponibilidade dos inerais no trato digestório. O perfil metabólico auxilia na avaliação de índices produtivos, para tanto, se faz análises de componentes bioquímicos do sangue. O objetivo foi estudar os efeitos que fontes orgânicas e inorgânicas de cobre e enxofre possuem nos parâmetros bioquímicos. O experimento foi realizado na FZEA/USP, para tanto 40 ovinos foram distribuídos em 10tratamentos: 1) dieta basal; 2) dieta contendo Mo; 3) dieta basal+ Cu inorg + S inorg; 4) dieta basal + Cu inorg + S org; 5) dieta basal + Cu org + S inorg; 6) dieta basal + Cu org + S org; 7) dieta com Mo + Cu inorg + S inorg; 8) dieta com Mo + Cu inorg + S org; 9) dieta com Mo + Cu org + S inorg; 10) dieta com Mo + Cu org + S org. De acordo com cada tratamento houve a inclusão de 10mgkg-1 de MS de Cu inorgânico ou orgânico ou 10 mg kg-1 de MS de Mo ou 0,2% de S inorgânico ou orgânico. Os animais receberam dieta única duas vezes ao dia com inclusão de volumoso num total de 3% do peso vivo. O experimento teve duração de 84 dias, com coletas a cada 28 dias para estudo de glicose, ureia, albumina, colesterol, triglicerídeos. Os parâmetros foram analisados com estrutura fatorial 2 x 2 x 2 (com e sem Mo, Cu orgânico e inorgânico e S orgânico e inorgânico) e uma dieta basal e uma basal mais molibdênio, em delineamento inteiramente casualizado, com um nível de significância de 5%. Os teores séricos de glicose, ureia, albumina e colesterol não apresentaram diferença significativa entre tratamentos e tempo, tão pouco foram influenciados (p>0.05) pelos tratamentos, sendo as médias 64,8mg dl-1; 30,0mg dl-1; 2,78mg L-1; 72,2mg dl-1respectivamente, apresentando-se dentro da normalidade. Os teores de triglicerídeos nos tratamentos (28.8; 34.8; 30.8; 36.9; 34.3; 27.0; 31.6; 32.1; 34.6; 31.1mg dl-1) foram influenciados pela interação Cu x S.

Structural basis for the pathophysiology of lipoprotein(a) in the athero-thrombotic process

Lipoprotein Lp(a) is a major and independent genetic risk factor for atherosclerosis and cardiovascular disease. The essential difference between Lp(a) and low density lipoproteins (LDL) is apolipoprotein apo(a), a glycoprotein structurally similar to plasminogen, the precursor of plasmin, the fibrinolytic enzyme. This structural homology endows Lp(a) with the capacity to bind to fibrin and to membrane proteins of endothelial cells and monocytes, and thereby to inhibit plasminogen binding and plasmin generation. The inhibition of plasmin generation and the accumulation of Lp(a) on the surface of fibrin and cell membranes favor fibrin and cholesterol deposition at sites of vascular injury. Moreover, insufficient activation of TGF-ß due to low plasmin activity may result in migration and proliferation of smooth muscle cells into the vascular intima. These mechanisms may constitute the basis of the athero-thrombogenic mode of action of Lp(a). It is currently accepted that this effect of Lp(a) is linked to its concentration in plasma. An inverse relationship between Lp(a) concentration and apo(a) isoform size, which is under genetic control, has been documented. Recently, it has been shown that inhibition of plasminogen binding to fibrin by apo(a) is also inversely associated with isoform size. Specific point mutations may also affect the lysine-binding function of apo(a). These results support the existence of functional heterogeneity in apolipoprotein(a) isoforms and suggest that the predictive value of Lp(a) as a risk factor for vascular occlusive disease would depend on the relative concentration of the isoform with the highest affinity for fibrin

Apolipoprotein and lipid abnormalities in chronic liver failure

Total serum lipids, as well as apolipoproteins A-I (apo A-I) and B (apo B), were determined in 74 patients with chronic liver failure without cholestasis and in 82 normal subjects. The VLDL, LDL and HDL lipid fractions were reduced in the liver failure group by 36%, 24% and 46%, respectively (P<0.001). Apolipoproteins A-I and B were also reduced by 26% and 25%, respectively (P<0.001). However, the reduction of HDL cholesterol (HDLc) was more pronounced than that of apo A-I and the HDLc:apo A-I ratio was significantly lower in the liver failure group. After separating these patients into groups with plasma albumin lower than 3.0, between 3.0 and 3.5, and higher than 3.5 g/dl, the HDLc:apo A-I ratio was proportional to plasma albumin, but the correlation was not statistically significant. When these patients were separated by the Child classification of liver function, there was a correlation between the HDLc:apo A-I ratio and liver function. The differences in the HDLc:apo A-I ratio between the Child groups B and C, and A and C were statistically significant (P<0.05). We conclude that there is a more pronounced reduction in HDL cholesterol than in apo A-I in liver failure patients. Therefore, the HDLc:apo A-I ratio is a marker of liver function, probably because there is a decreased lecithin-cholesterol acyltransferase production by the diseased liver

The Lebanese mutation as an important cause of familial hypercholesterolemia in Brazil

Familial hypercholesterolemia (FH) is a common autosomal disorder that affects about one in 500 individuals in most Western populations and is caused by a defect in the low-density-lipoprotein receptor (LDLr) gene. In this report we determined the molecular basis of FH in 59 patients from 31 unrelated Brazilian families. All patients were screened for the Lebanese mutation, gross abnormalities of the LDLr gene, and the point mutation in the codon 3500 of the apolipoprotein B-100 gene. None of the 59 patients presented the apoB-3500 mutation, suggesting that familial defective ApoB-100 (FDB) is not a major cause of inherited hypercholesterolemia in Brazil. A novel 4-kb deletion in the LDLr gene, spanning from intron 12 to intron 14, was characterized in one family. Both 5' and 3' breakpoint regions were located within Alu repetitive sequences, which are probably involved in the crossing over that generated this rearrangement. The Lebanese mutation was detected in 9 of the 31 families, always associated with Arab ancestry. Two different LDLr gene haplotypes were demonstrated in association with the Lebanese mutation. Our results suggest the importance of the Lebanese mutation as a cause of FH in Brazil and by analogy the same feature may be expected in other countries with a large Arab population, such as North American and Western European countries.

HFE gene mutations in coronary atherothrombotic disease

Although iron can catalyze the production of free radicals involved in LDL lipid peroxidation, the contribution of iron overload to atherosclerosis remains controversial. The description of two mutations in the HFE gene (Cys282Tyr and His63Asp) related to hereditary hemochromatosis provides an opportunity to address the question of the association between iron overload and atherosclerosis. We investigated the prevalence of HFE mutations in 160 survivors of myocardial infarction with angiographically demonstrated severe coronary atherosclerotic disease, and in 160 age-, gender- and race-matched healthy control subjects. PCR amplification of genomic DNA followed by RsaI and BclI restriction enzyme digestion was used to determine the genotypes. The frequency of the mutant Cys282Tyr allele was identical among patients and controls (0.022; carrier frequency, 4.4%), whereas the mutant His63Asp allele had a frequency of 0.143 (carrier frequency, 27.5%) in controls and of 0.134 (carrier frequency, 24.5%) in patients. Compound heterozygotes were found in 2 of 160 (1.2%) controls and in 1 of 160 (0.6%) patients. The finding of a similar prevalence of Cys282Tyr and His63Asp mutations in the HFE gene among controls and patients with coronary atherothrombotic disease, indirectly questions the possibility of an association between hereditary hemochromatosis and atherosclerosis.

Eggplant (Solanum melongena) infusion has a modest and transitory effect on hypercholesterolemic subjects

Eggplant (Solanum melongena) is consumed extensively in Brazil. It has been believed that infusion of a powdered preparation of the fruit may reduce serum cholesterol. However, there are few documented reports on its effects on cholesterol metabolism and its possible hypocholesterolemic effect has not been proved by well-controlled studies. The aim of the present study was to observe the effects of S. melongena on the serum cholesterol and triglycerides of 38 hypercholesterolemic human volunteers ingesting S. melongena infusion for five weeks. Thirty-eight hypercholesterolemic subjects receiving either S. melongena infusion (N = 19) or placebo (N = 19) participated in two clinical experiments in which the effect of S. melongena infusion was studied with (N = 16) or without (N = 38) dietary orientation. Total cholesterol and its fractions, triglycerides, and apolipoproteins A and B were measured in blood at the beginning of the experiment and three and five weeks thereafter. No differences were observed compared to control. Intraindividual analysis showed that S. melongena infusion significantly reduced the blood levels of total and LDL cholesterol and of apolipoprotein B. After dietary orientation, no intra- or intergroup differences were seen for any of the parameters analyzed. The results suggest that S. melongena infusion had a modest and transitory effect, which was not different from that obtained with standard orientation for dyslipidemia patients (diet and physical activities).

Polymorphisms of the low-density lipoprotein receptor gene in Brazilian individuals with heterozygous familial hypercholesterolemia

Familial hypercholesterolemia (FH) is a metabolic disorder inherited as an autosomal dominant trait characterized by an increased plasma low-density lipoprotein (LDL) level. The disease is caused by several different mutations in the LDL receptor gene. Although early identification of individuals carrying the defective gene could be useful in reducing the risk of atherosclerosis and myocardial infarction, the techniques available for determining the number of the functional LDL receptor molecules are difficult to carry out and expensive. Polymorphisms associated with this gene may be used for unequivocal diagnosis of FH in several populations. The aim of our study was to evaluate the genotype distribution and relative allele frequencies of three polymorphisms of the LDL receptor gene, HincII1773 (exon 12), AvaII (exon 13) and PvuII (intron 15), in 50 unrelated Brazilian individuals with a diagnosis of heterozygous FH and in 130 normolipidemic controls. Genomic DNA was extracted from blood leukocytes by a modified salting-out method. The polymorphisms were detected by PCR-RFLP. The FH subjects showed a higher frequency of A+A+ (AvaII), H+H+ (HincII1773) and P1P1 (PvuII) homozygous genotypes when compared to the control group (P<0.05). In addition, FH probands presented a high frequency of A+ (0.58), H+ (0.61) and P1 (0.78) alleles when compared to normolipidemic individuals (0.45, 0.45 and 0.64, respectively). The strong association observed between these alleles and FH suggests that AvaII, HincII1773 and PvuII polymorphisms could be useful to monitor the inheritance of FH in Brazilian families.

In vitro cytotoxicity of the LDE: daunorubicin complex in acute myelogenous leukemia blast cells

Acute myelogenous leukemia (AML) blast cells show high-affinity degradation of low-density lipoprotein (LDL), suggesting an increased expression of cellular LDL receptors. LDE is a lipid microemulsion easily synthesized in vitro which is known to mimic the metabolic pathway of LDL. We used LDE as a carrier for daunorubicin and assayed the cytotoxicity of the complex using AML blast cells since RT-PCR analysis showed that AML cells express LDL receptor mRNA. The LDE:daunorubicin complex killed 46.7% of blast cells and 20.2% of normal bone marrow cells (P<0.001; Student t-test). Moreover, this complex destroyed AML blast cells as efficiently as free daunorubicin. Thus, LDE might be a suitable carrier of chemotherapeutic agents targeting these drugs to neoplastic cells and protecting normal tissues.

Estrogen replacement therapy and cardioprotection: mechanisms and controversies

Epidemiological and case-controlled studies suggest that estrogen replacement therapy might be beneficial in terms of primary prevention of coronary heart disease (CHD). This beneficial effect of estrogens was initially considered to be due to the reduction of low density lipoproteins (LDL) and to increases in high density lipoproteins (HDL). Recent studies have shown that estrogens protect against oxidative stress and decrease LDL oxidation. Estrogens have direct effects on the arterial tissue and modulate vascular reactivity through nitric oxide and prostaglandin synthesis. While many of the effects of estrogen on vascular tissue are believed to be mediated by estrogen receptors alpha and ß, there is evidence for `immediate non-genomic' effects. The role of HDL in interacting with 17ß-estradiol including its esterification and transfer of esterified estrogens to LDL is beginning to be elucidated. Despite the suggested positive effects of estrogens, two recent placebo-controlled clinical trials in women with CHD did not detect any beneficial effects on overall coronary events with estrogen therapy. In fact, there was an increase in CHD events in some women. Mutations in thrombogenic genes (factor V Leiden, prothrombin mutation, etc.) in a subset of women may play a role in this unexpected finding. Thus, the cardioprotective effect of estrogens appears to be more complicated than originally thought and requires more research.

Six years of treatment with the HELP system of a patient with familial hypercholesterolemia

The purpose of the present report is to demonstrate the long-term efficacy and safety of heparin-induced extracorporeal lipoprotein precipitation (HELP) of LDL-c and fibrinogen in the management of familial hypercholesterolemia. From June 1992 to June 1998 a 22-year-old young male patient with familial hypercholesterolemia (double heterozygote for C660X and S305C) resistant to medication and diet and with symptomatic coronary artery disease (angina) was treated weekly with 90-min sessions of the HELP system. The patient had also been previously submitted to right coronary artery angioplasty. The efficacy of the method was evaluated by comparing the reduction of total cholesterol, LDL-c and fibrinogen before and after the sessions and before and after initiation of the study (data are reported as averages for each year). During the study, angina episodes disappeared and there were no detectable adverse effects of the treatment. Total cholesterol (TC), fibrinogen, and LDL-c decreased significantly after each session by 59.6, 66.1 and 64%, respectively. HDL-c showed a nonsignificant reduction of 20.4%. Comparative mean values pre- and post-treatment values in the study showed significant differences: TC (488 vs 188 mg/dl), LDL-c (416.4 vs 145 mg/dl), and fibrinogen (144.2 vs 57.4 mg/dl). There was no significant change in HDL-c level: 29.4 vs 23 mg/dl. These data show that the HELP system, even for a long period of time, is a safe and efficient mode of treatment of familial hypercholesterolemia and is associated with disappearance of angina symptoms.

Nitric oxide, cholesterol oxides and endothelium-dependent vasodilation in plasma of patients with essential hypertension

The objective of the present study was to identify disturbances of nitric oxide radical (·NO) metabolism and the formation of cholesterol oxidation products in human essential hypertension. The concentrations of·NO derivatives (nitrite, nitrate, S-nitrosothiols and nitrotyrosine), water and lipid-soluble antioxidants and cholesterol oxides were measured in plasma of 11 patients with mild essential hypertension (H: 57.8 ± 9.7 years; blood pressure, 148.3 ± 24.8/90.8 ± 10.2 mmHg) and in 11 healthy subjects (N: 48.4 ± 7.0 years; blood pressure, 119.4 ± 9.4/75.0 ± 8.0 mmHg).Nitrite, nitrate and S-nitrosothiols were measured by chemiluminescence and nitrotyrosine was determined by ELISA. Antioxidants were determined by reverse-phase HPLC and cholesterol oxides by gas chromatography. Hypertensive patients had reduced endothelium-dependent vasodilation in response to reactive hyperemia (H: 9.3 and N: 15.1% increase of diameter 90 s after hyperemia), and lower levels of ascorbate (H: 29.2 ± 26.0, N: 54.2 ± 24.9 µM), urate (H: 108.5 ± 18.9, N: 156.4 ± 26.3 µM), ß-carotene (H: 1.1 ± 0.8, N: 2.5 ± 1.2 nmol/mg cholesterol), and lycopene (H: 0.4 ± 0.2, N: 0.7 ± 0.2 nmol/mg cholesterol), in plasma, compared to normotensive subjects. The content of 7-ketocholesterol, 5alpha-cholestane-3ß,5,6ß-triol and 5,6alpha-epoxy-5alpha-cholestan-3alpha-ol in LDL, and the concentration of endothelin-1 (H: 0.9 ± 0.2, N: 0.7 ± 0.1 ng/ml) in plasma were increased in hypertensive patients. No differences were found for ·NO derivatives between groups. These data suggest that an increase in cholesterol oxidation is associated with endothelium dysfunction in essential hypertension and oxidative stress, although ·NO metabolite levels in plasma are not modified in the presence of elevated cholesterol oxides.