Differences in adiponectin protein expression: effect of fat depots and type 2 diabetic status.

Adiponectin is an adipocyte-derived hormone associated with insulin sensitivity and atherosclerotic risk. As central rather than gluteofemoral fat is known to increase the risk of type 2 diabetes and cardiovascular disease, we investigated the mRNA and protein expression of adiponectin in human adipose tissue depots. RNA was extracted from 46 human adipose tissue samples from non-diabetic subjects aged 44.33 +/- 12.4 with a BMI of 28.3 +/- 6.0 (mean +/- SD). The samples were as follows: 21 abdominal subcutaneous, 13 omentum, 6 thigh; samples were also taken from diabetic subjects aged 66.6 +/- 7.5 with BMI 28.9 +/- 3.17; samples were: 6 abdominal subcutaneous; 3 thigh. Quantitative PCR and Western analysis was used to determine adiponectin content. Protein content studies determined that when compared with non-diabetic abdominal subcutaneous adipose tissue (Abd Sc AT) (values expressed as percentage relative to Abd Sc AT -100 %). Adiponectin protein content was significantly lower in non-diabetic omental AT (25 +/- 1.6 %; p < 0.0001, n = 6) and in Abd Sc AT from diabetic subjects (36 +/- 1.5 %; p < 0.0001, n = 4). In contrast, gluteal fat maintained high adiponectin protein content from non-diabetic patients compared with diabetic patients. An increase in BMI was associated with lower adiponectin protein content in obese ND Abd Sc AT (25 +/- 0.4 %; p < 0.0001). These findings were in agreement with the mRNA expression data. In summary, this study indicates that adiponectin protein content in non-diabetic subjects remains high in abdominal subcutaneous fat, including gluteal fat, explaining the high serum adiponectin levels in these subjects. Omental fat, however, expresses little adiponectin. Furthermore, abdominal and gluteal subcutaneous fat appears to express significantly less adiponectin once diabetic status is reached. In conclusion, the adipose tissue depot-specific expression of adiponectin may influence the pattern of serum adiponectin concentrations and subsequent disease risk.

[Influence of planting density and precipitation on N2O emission from a winter wheat field].

To investigate the impact of plant density on N2O emission from winter wheat field and the cause of seasonal variation in the emission, field experiment with four planting rates of 0, 90, 180 and 270 kg/ha was conducted at the Jiangning County near Nanjing during 1999-2000 winter wheat growing season. Data of the field measurements indicated that the N2O emission rates during the season from planting to overwintering were not influenced by the plant density, while the emission was positively correlated with the planting density during the season from turning green to maturity. The emissions from the field plots with planting rates of 0 and 90 kg/ha were not found to be significantly different. A further analysis suggested that the seasonal variation of N2O emission be mainly influenced by precipitation, which could be quantitatively described by an exponential function of a weighted average precipitation of 6-day period before measurement.

Quantitative determination of inositol in Hymenolepis diminuta (Cestoda).

Myoinositol and scylloinositol have been identified qualitatively and quantitatively by gas-liquid chromatography in Hymenolepis diminuta. No myoinosose-2 was detected. Myoinositol was unevenly distributed throughout the worm. The scolex and neck regions contained more free- and phosphatidyl-bound inositol. This region also contained more lipid-bound phosphorus, but less total lipid and water.

Membrane transport of inositol by Hymenolepis diminuta (Cestoda).

The absorption of myoinositol by Hymenolepis diminuta involved diffusion that occurred at all substrate concentrations tested; at low substrate concentrations the mediated component predominated. The mediated process exhibited saturation kinetics with a Vmax and Kt of 0.011 mumoles/g of the ethanol-extracted dry wt/4 min and 0.0067 mM, respectively. Myoinositol transport was sensitive to changes in temperature, pH, and sodium ion concentration. D-Glucose was a noncompetitive inhibitor of myoinositol transport, but myoinositol had no effect on D-glucose absorption. Phlorizin interacted competitively with the myoinositol transport system. Various sugar alcohols and amino acids had no effect on myoinositol transport. These results indicate that a separate transport system exists in H. diminuta for myoinositol.

Carbohydrate transport in Moniliformis dubius (Acanthocephala). III. Post-absorptive fate of fructose, mannose, and galactose.

The initial metabolism of fructose, mannose, and galactose in Moniliformis dubius (M. moniliformis; Acanthocephala) was examined following brief aerobic incubations in 14C-labeled substrate. The pattern of incorporation of radioactivity from 14C-fructose and 14C-mannose into intermediates of hexose metabolism was indistinguishable from that reported elsewhere for the initial post-absorptive metabolism of 14C-glucose under comparable conditions; these hexoses were phosphorylated rapidly following their absorption, and much of the radioactivity absorbed as mannose or fructose was recovered in the nonreducing disaccharide trehalose [alpha-D-glucopyranosyl-(1 leads to 1)-alpha-D-glucopyranoside]. 14C-Galactose was phosphorylated less readily than the other hexoses. More than half of the radioactivity absorbed as galactose was incorporated into disaccharide; some of the galactose-derived disaccharide had properties suggesting the presence of a galactosyl moiety. Incorporation of radioactivity from any of the hexoses into glycogen was minimal. The extensive incorporation of hexose moieties into trehalose or a trehalose-like disaccharide rather than glycogen underscores the probable importance of trehalose metabolism to carbohydrate assimilation in Moniliformis. Physiological factors which might favor trehalose biosynthesis over glycogenesis are considered.

Cytofluorograf detection of Plasmodium yoelii, Trypanosoma gambiense, and Trypanosoma equiperdum by laser excited fluorescence of stained rodent blood.

Samples of rat blood infected with Plasmodium yoelii (3% parasitized erythrocytes), Trypanosoma gambiense, or Trypanosoma equiperdum (greater than 50 trypanosomes per microscope field at 400 X) were fixed with 0.5% glutaraldehyde in phosphate buffered saline, then stained with acridine orange (AO) at 10(-4), 10(-5), or 10(-6) M for 0 to 15 min at 5 C or 25 C and/or ethidium bromide (EB) at 0.05 mg/ml for 20 min at 25 C. Stained cells were analyzed with a laser Cytofluorograf (Bio/Physics Systems, Inc.) to determine if parasites could be detected and differentiated from blood cells by their fluorescent characteristics. Samples of uninfected rat blood with and without leukocytes and P. yoelii-infected blood without leukocytes were treated similarly. In addition, suspensions of T. gambiense and T. equiperdum without all blood cells were stained with AO or EB and analyzed with the Cytofluorograf, as were mixed suspensions of both trypanosome species. EB- but not AO-stained P. yoelii-infected erythrocytes had fluorescent characteristics different from most blood cells. Neither AO- nor EB-stained T. gambiense or T. equiperdum could be differentiated from host blood cells or from each other. The results are discussed with respect to the use of laser flow systems in the detection and analysis of bloodstream dwelling protozoan parasites.

Carbohydrate effects of amino acid transport by Trypanosoma equiperdum.

Uptake of 14C-labeled alanine, glutamate, lysine, methionine, proline, and phenylalanine by Trypanosoma equiperdum during 2-minute incubations occurred by diffusion and membrane-mediated processes. Amino acid metabolism was not detected by paper chromatography of trypanosome extracts. Most of 18 carbohydrates tested for ability to alter amino acid transport neither changed nor significantly inhibited transport. Glucose, however, stimulated glutamate, lysine and proline transport; fructose stimulated lysine uptake and 2-deoxy-D-glucose increased phenylalanine and methionine absorption. No evidence was found that the carbohydrates acted by binding to amino acid transport "sites." Glucose inhibition of alanine, phenylalanine, and methionine uptake was linked to glycolysis. The rapid formation of alanine form glucose stimulated alanine release and, when glycolysis was blocked, glucose no longer inhibited alanine transport. Methionine and phenylalanine release was also stimulated by glucose. Glucose changed the ability of lysine, glutamate, and proline to inhibit each others' uptake, indicating that certain amino acids are preferentially absorbed by respiring cells. Analysis of free pool amino acid levels suggested that some amino acid transport systems in T. equiperdum are linked in such a way to glycolysis as to control the cell concentrations of these amino acids.

Carbohydrate transport in Moniliformis dubius (Acanthocephala). I. The kinetics and specificity of hexose absorption.

The uptakes of 14C-glucose, -2-deoxyglucose, -mannose, -N-acetylglucosamine, -3-0-methylglucose, -fructose, and -galactose by female Moniliformis dubius were nonlinear, saturable functions of hexose concentration. Kinetic and inhibition studies indicated that glucose and 2-deoxyglucose were absorbed via a single common transport locus. Mannose, N-acetylglucosamine, 3-0-methylglucose, fructose, and galactose (in decreasing order of effectiveness) inhibited the uptake of glucose in a completely competitive manner; their absorptions appeared to be mediated by the glucose transport locus and, to some degree, by one or more additional transport systems. Kinetic studies suggested that the apparent inhibitions of 14C-glucose uptake by maltose and glucose-6-phosphate were due to free glucose liberated through the action of surface hydrolases. The uptake of 14C-glucose was also inhibited by salicin, alpha-methylglucoside, and beta-methylglucoside, but not by pentoses, L-hexoses, sugar alcohols, disaccharides (except maltose), gluconic acid, glucuronic acid, phlorizin, or ouabain. Glucose uptake was not Na+-dependent.