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PubMed2006

A combined 1H-NMR spectroscopy- and mass spectrometry-based metabolomic study of the PPAR-alpha null mutant mouse defines profound systemic changes in metabolism linked to the metabolic syndrome.

The mobilization of triacylglycerides from storage in adipocytes to the liver is a vital response to the fasting state in mammalian metabolism. This is accompanied by a rapid translational activation of genes encoding mitochondrial, microsomal, and peroxisomal beta-oxidation in the liver, in part under the regulation of peroxisome proliferator-activated receptor-alpha (PPAR-alpha). A failure to express PPAR-alpha results in profound metabolic perturbations in muscle tissue as well as the liver. These changes represent a number of deficits that accompany diabetes, dyslipidemia, and the metabolic syndrome. In this study, the metabolic role of PPAR-alpha has been investigated in heart, skeletal muscle, liver, and adipose tissue of PPAR-alpha null mice at 1 mo of age using metabolomics. To maximize the coverage of the metabolome in these tissues, (1)H-NMR spectroscopy, magic angle spinning (1)H-NMR spectroscopy, gas chromatography-mass spectrometry, and liquid chromatography-mass spectrometry were used to examine metabolites in aqueous tissue extracts and intact tissue. The data were analyzed by the multivariate approaches of principal components analysis and partial least squares. Across all tissues, there was a profound decrease in glucose and a number of amino acids, including glutamine and alanine, and an increase in lactate, demonstrating that a failure to express PPAR-alpha results in perturbations in glycolysis, the citric acid cycle, and gluconeogenesis. Furthermore, despite PPAR-alpha being weakly expressed in adipose tissue, a profound metabolic perturbation was detected in this tissue.

Adipose Tissue↗
PubMed1981

Characterization of somatostatin-like components in the tumors and plasma of a patient with a somatostatinoma.

Somatostatin-like immunoreactive polypeptides with approximate molecular weights of 1,500-2,000 (peak I), 2,500-3,500 (peak II), and 10,000-15,000 (peak III) were isolated from the peripheral plasma and from extracts of tumor tissue of a patient with a pancreatic somatostatinoma and hepatic metastases. The peak I component had approximately the same molecular charge as somatostatin, but the peak II component was appreciably more basic. The peak III component was heterogeneous with respect to charge and dissociated into smaller immunoreactive polypeptides during isoelectric focusing or after treatment with strongly denaturing solvents. In plasma, the levels of the larger molecular weight components (peak II and peak III), relative to that of the somatostatin-size component (peak I), were higher than in tissue extracts, suggesting a lower rate of clearance of the larger polypeptides from the circulation. In response to a nutrient stimulus, the peak I component increased to a greater extent than the larger components. Plasma levels of larger immunoreactive polypeptides relative to the peak I component in patient's plasma were higher after complete resection of the pancreatic tumor than preoperatively, suggesting that the rate of release of these putative precursor forms from the metastases was greater than from the primary tumor.

Chemical Phenomena↗
PubMed1989

Human placental sterylsulfatase: immunocytochemical and biochemical localization.

Human placental sterylsulfatase was localised in situ by light and electron microscope immunocytochemical techniques as well as in homogenate and tissue extract fractions by enzyme assays. Light microscope observations on frozen sections of term and preterm placenta revealed sterylsulfatase immunoactivity primarily in the syncytiotrophoblast. Electron microscope observations confirmed the light microscope findings; in addition, they showed that the sulfatase is present in the endoplasmic reticulum of endothelial cells, too. In the syncytiotrophoblast, the enzyme was detectable in the cytoplasmic membrane of the nuclear evelope, in the membranes of the rough endoplasmic reticulum, in the plasma membrane with predominant localisation in coated pits, and in the membranes of endosomes and multivesicular bodies; little or no reactivity was detectable over the membranes of the Golgi complex and of lysosomes. Sterylsulfatase immunoactivity was absent in placentas with hereditary sterylsulfatase deficiency. The observations indicate that human placental sterylsulfatase is normally present in the membranes of compartments along the secretory pathway and the endocytic route of cells lining the fetal and maternal blood. Homogenates of normal term placenta as well as membrane vesicle preparations obtained by extraction of trophoblast tissue with isotonic saline were fractionated by differential centrifugation; the fractions were assayed for specific activities of sterylsulfatase and several marker enzymes of cellular topography. In agreement with our immunocytochemical findings, the results of these biochemical localisation experiments indicate the repeatedly described association of the placental sterylsulfatase with microsomal membranes but also point to the presence of the enzyme's activity in the microvillous plasma membrane of the syncytiotrophoblast. This localisation of sterylsulfatase may have functional implications in the placental uptake of circulating steroid sulfates.

Arylsulfatases↗
PubMed2005

Evaluation of the inhibitory effect of triphala on PMN-type matrix metalloproteinase (MMP-9).

BACKGROUND: This study evaluated the inhibitory activity of triphala on PMN-type matrix metalloproteinase (MMP-9) expressed in adult periodontitis patients and compared its activity with another ayurvedic drug, kamillosan, and doxycycline, which has known inhibitory activity. METHODS: Matrix metalloproteinases (MMPs) were extracted from gingival tissue samples from 10 patients (six males, four females) with chronic periodontitis. Tissue extracts were treated with the drug solutions, the inhibition was analyzed by gelatin zymography, and the percentage of inhibition was determined by a gel documentation system. RESULTS: The activity of MMPs was significantly decreased with the use of the drugs. Triphala showed a 76.6% reduction of MMP-9 activity, whereas kamillosan showed a 46.36% reduction at a concentration of 1,500 microg/ml (crude extract) and doxycycline showed a 58.7% reduction at a concentration of 300 microg/ml (pure drug). CONCLUSION: The present study showed the strong inhibitory activity of triphala on PMN-type MMPs involved in the extracellular matrix (ECM) degradation during periodontitis.

Adult↗
PubMed2000

Acute oxygen supplementation restores markers of hepatocyte energy status and hypoxia in cirrhotic rats.

The oxygen limitation hypothesis states that hepatocyte hypoxia is the mechanism determining metabolic restriction in the cirrhotic liver. Therefore we studied markers of hepatocyte energy state and cellular hypoxia in livers of normal and cirrhotic rats before and after oxygen supplementation. Rats with carbon tetrachloride-induced cirrhosis and procedural control rats were exposed to either room air or a hyperoxic gas mixture for 1 h immediately before freeze clamping and perchloric acid extraction of liver tissue. Extracts were assessed by (31)P NMR and enzymatic assays. Livers from cirrhotic rats breathing room air showed a reduced ratio of ATP/ADP, an increased ratio of inorganic phosphate/ATP, and a trend toward an increased ratio of lactate/pyruvate compared with procedural control livers (ATP/ADP 1.73 +/- 0.35 versus 2.68 +/- 0.61, P <.05; P(i)/ATP 2.74 +/- 0.48 versus 1.56 +/- 0.26, P <.05; lactate/pyruvate 29.3 +/- 6.4 versus 22.5 +/- 7.4, P =.18). After supplementation with oxygen for 1 h, these ratios in cirrhotic livers approached control values. A variety of other metabolic markers affected by cirrhosis showed variable trends toward normal in response to oxygen supplementation, whereas minor trends toward an increase in ATP levels in control animals suggest the possibility of marginal oxygen limitation in normal livers. The data are consistent with the hypothesis that hepatocytes in cirrhotic livers have normal metabolic capacity but are constrained by a deficit in oxygen supply. Interventions aimed at increasing oxygen supply to the liver may have both short- and long-term therapeutic value in the management of cirrhosis.

Animals↗
PubMed1975

A screening method for sulfonamides extracted from animal tissues.

A screening method for the estimation of possible residues of sulfonamides in poultry tissues is described. The method utilizes thin layer chromatography (TLC) to separate Bratton-Marshall positive reactants. In the absence of interference and the identification of 1 sulfonamide by TLC, the colorimetric method is recommended for quantitation. When interferences are present, TLC should be used for both qualitative and quantitative analysis. The screening method has a sensitivity of less than 0.05 ppm and a recovery of greater than 80%.

Animals↗
PubMed1990

[The mechanism of natural resistance in the rat to Entamoeba histolytica].

The data presented in this paper may be summarized as follows: 1) intraportal injection of a virulent strain (HM-1) of Entamoeba histolytica in Wistar rats, of both sexes, gives way to non-progressive microscopic changes, characterized by a rapid leukocytic reaction surrounding the amebas, disappearance of the parasites within 5 hours, and total lack of hepatic damage; 2) leucopenia only modifies the previous description by the fact that there are no leucocytes around the amebas, although these disappear in the same time, showing, at this moment, an early and prominent vacuolar degeneration; 3) hypocomplementaemia shows the same results as leucopenia; 4) fragments and extracts from various tissues from the rat and hamster show variable degrees of interference with the viability of axenic amebas of Entamoeba histolytica conserved in culture; only a minimal part of the interference to be due to the activity of the complement which is present in the tissue extracts.

Animals↗
PubMed1990

Lactate dehydrogenase and creatine phosphokinase isoenzymes in tissues of laboratory animals.

The lactate dehydrogenase (LDH) and creatine phosphokinase (CPK) isoenzyme distributions in tissues of the ICR mouse, Wistar rat, guinea pig and golden hamster were analyzed by histoelectrophoresis. Tissues obtained were as follows: liver, pancreas, stomach, small intestine, heart, femoral muscle, uterus, kidney, spleen, lymph node, cerebrum, spinal cord and erythrocyte. Histoelectrophoresis was for the direct analysis of LDH and CPK isoenzymes in the tissues and had high practical value compared with previous tissue-extraction methods. In tissues of the mouse, guinea pig and golden hamster, LDH isoenzymes showed five bands. In the rat, LDH isoenzyme was separated into four fractions. CPK isoenzyme showed three bands; BB, MB and MM. In some tissues, the MM band was separated into two sub fractions.

Animals↗