Hexaclorobenceno y pentacloro fenol en tejido adiposo y orina de un nino con porfiria cutanea tardia: evaluacion del tratamiento(s-adenosil-L-metionina y bajas dosis de cloriquina): a cuatro anos de inicido el mismo / Hexaclorobenzene and poentachlorophenol in fatty tissue and urine of child with porfiria cutanea tarda. Treatment evaluation (S-Adenosyl-L-Methionine and low dose of Chloroquine) four years after

Se ha estudiado el contenido de hexaclorobenceno(HCB) y pentaclorofenol(PCP)en tejido adiposo y orina de un ninio con porfiria cutanea tardia (PCT) hereditaria y del producto quimico con el cual estuvo en contacto. El contenido de HCB (2,3 ug/g) en orinayPCF (5,0 ml) fueron elevados respectoa los valores normales (HCB tej. adiposo<1 ug/gyPCF urinario ,0,1 ng/ml). L presencia de PCF en orina,indicador de intoxicacion por HCB,esta sugiriendo que este policlorado fue el desencadenante de la PCT en este ninio portador de la enfermedad. La variacion del contenido de porfirinas urinarias durante los tratamientos con A)bicarbonato (10mg/dia)y B) conbinado:S adenosil-L-Metionina(SAM)(12mg/kg/diax20dias) y cloroquina(2x110mg/semana) mostraron: En A)disminucion del 50 por ciento de las porfirinas iniciales (1700 ug/24h) luego de 50 meses de tratamiento,B) disminucion hasta alcanzar los valores normales(20-250 ug/24 h) luego de 4 meses de tratamiento. Las porfirinas se mantuvieron en valores normalesaun hasta 4 anios de iniciada la terapia con SAM y cloriquina

Hexaclorobenceno y pentacloro fenol en tejido adiposo y orina de un nino con porfiria cutanea tardia: evaluacion del tratamiento(s-adenosil-L-metionina y bajas dosis de cloriquina): a cuatro anos de inicido el mismo / Hexaclorobenzene and poentachlorophenol in fatty tissue and urine of child with porfiria cutanea tarda. Treatment evaluation (S-Adenosyl-L-Methionine and low dose of Chloroquine) four years after

Se ha estudiado el contenido de hexaclorobenceno(HCB) y pentaclorofenol(PCP)en tejido adiposo y orina de un ninio con porfiria cutanea tardia (PCT) hereditaria y del producto quimico con el cual estuvo en contacto. El contenido de HCB (2,3 ug/g) en orinayPCF (5,0 ml) fueron elevados respectoa los valores normales (HCB tej. adiposo<1 ug/gyPCF urinario ,0,1 ng/ml). L presencia de PCF en orina,indicador de intoxicacion por HCB,esta sugiriendo que este policlorado fue el desencadenante de la PCT en este ninio portador de la enfermedad. La variacion del contenido de porfirinas urinarias durante los tratamientos con A)bicarbonato (10mg/dia)y B) conbinado:S adenosil-L-Metionina(SAM)(12mg/kg/diax20dias) y cloroquina(2x110mg/semana) mostraron: En A)disminucion del 50 por ciento de las porfirinas iniciales (1700 ug/24h) luego de 50 meses de tratamiento,B) disminucion hasta alcanzar los valores normales(20-250 ug/24 h) luego de 4 meses de tratamiento. Las porfirinas se mantuvieron en valores normalesaun hasta 4 anios de iniciada la terapia con SAM y cloriquina

Glucocorticoid receptor messenger ribonucleic acid in different regions of human adipose tissue.

The expression of glucocorticoid receptor (GR) messenger RNA (mRNA) was investigated in sc adipose tissue and isolated adipocytes from the abdominal and gluteal regions in men and women using a human GR complementary RNA probe. GR mRNA levels were 2-fold higher in female than in male abdominal tissue or adipocytes, whereas in gluteal tissue or adipocytes no sex differences were observed. GR mRNA levels in female abdominal adipocytes were 50% higher than in corresponding female gluteal adipocytes; the opposite was observed corresponding in males. Northern blot analysis of total cellular RNA isolated from abdominal and gluteal adipocytes showed hybridization of the human GR probe to an RNA species of approximately 7.1 kilobases in both regions. No sex or regional differences in GR mRNA stability were observed. The human metallothionein II (hMTII) mRNA, which is regulated by glucocorticoids at the transcriptional level, showed an opposite sex and regional pattern as GR mRNA. However, in gluteal adipose tissue no sex differences were observed in hMTII mRNA levels. The expression of beta-actin mRNA, which is not regulated by glucocorticoids, showed no sex or regional variation. By immunocytochemistry, using an anti-GR-monoclonal antibody, cytoplasmic as well as nuclear staining for GR was demonstrated in both sexes and both regions. In conclusion, variations in GR mRNA levels between sexes and body regions may explain the well known sex and tissue differences in effects of glucocorticoids on human adipose tissue.

Pedersen fetuin contains three adipogenic factors with distinct biochemical characteristics.

Crude Pedersen fetuin, derived from fetal bovine serum, contains adipogenic activity. Biochemical characterization was undertaken by following the differentiation of the 1246 adipogenic cell line. The present paper provides evidence that crude fetuin contains distinct proteins with adipogenic activity. By molecular sieve fractionation using Sephacryl S-300, the majority of adipogenic activity eluted in two distinct peaks, FI (molecular weight greater than 669 kDa) and FII (molecular weight ranging from 445 and 232 kDa). In addition a minor activity was found in a third peak, FIII (molecular weight around 69 kDa). Partial purification and biochemical characterization indicate that FI and FII are two distinct factors. FI has a PI higher than 9.4, is destroyed by alkaline treatment, and is stable when treated with acid. FII has a PI lower than 4.0, is alkali stable, but is destroyed completely by treatment with acid. Moreover, our data show that adipogenic factors are distinct from another protein alpha 2 macroglobulin known to be found in crude Pedersen fetuin. These results suggest that serum contains two large molecular weight proteins bearing adipogenic activity which could play an important role in the control of the adipose differentiation process.

Subcutaneous adipose-tissue fatty acids and vitamin E in humans: relation to diet and sampling site.

The influence of diet and sampling site on subcutaneous adipose-tissue fatty acid composition and vitamin E content was examined in 20 healthy subjects. A dietary history and adipose-tissue biopsies from the buttock were obtained from 14 individuals. In another six individuals, samples were taken from both waist and buttock. The relative dietary intake of both polyunsaturated and saturated fatty acids correlated with the relative content in adipose tissue (r = 0.6, p = 0.02). Adipose-tissue vitamin E was strongly associated with dietary intake (r = 0.76, p = 0.004). The content of n - 3 (omega-3) fatty acids in adipose tissue was shown to influence adipose-tissue vitamin E negatively. Waist and buttock fat differed (p less than 0.05) with regard to fatty acid composition whereas no systematic variation was seen in the vitamin E content. The use of adipose-tissue biopsies in epidemiologic studies as measures of the habitual relative intake of fatty acids and vitamin E is suggested.

Purification of an apolipoprotein A binding protein from mouse adipose cells.

A protein recognizing apolipoproteins AI, AII and AIV was purified from cultured mouse adipose cells of the Ob17MT18 clonal line. Apolipoprotein A binding sites were solubilized in the presence of proteinase inhibitors using the non-denaturating detergent CHAPS. Chromatography of the soluble extract on DEAE-Trisacryl was followed by immunoaffinity chromatography of the complex apolipoprotein AI-binding proteins on anti-(apolipoprotein AI) coupled to Sepharose 4B and then by h.p.l.c. on an RP-Select B column. A 1400-fold purification over the starting crude homogenate was achieved. The purified material contained two proteins that were both able to bind apolipoproteins AI, AII and AIV, but not low-density lipoprotein. Glycopeptidase F treatment showed the existence of a single protein bearing either N-linked high-mannose or complex oligosaccharide chains. The purified material showed an apparent molecular mass of 80 +/- 9 kDa by h.p.l.c. on a TSKG 3000 SW column. Rabbit polyclonal antibodies directed against the purified material revealed two protein bands of 80 and 92 kDa after SDS/PAGE under reducing conditions and immunoblotting. These bands were undetectable in growing Ob17PY cells previously shown not to bind the various apolipoproteins A and not to undergo cholesterol efflux, whereas they were conspicuous in growth-arrested Ob17PY cells which have recovered these properties.

PCB reduction and clinical improvement by detoxification: an unexploited approach?

1. A detoxification trial was administered to a female worker from a capacitor factory who had been exposed to polychlorinated biphenyls (PCBs) and other lipophilic industrial chemicals. 2. The patient presented with severe abdominal complaints, chloracne, liver abnormalities, and a spontaneous nipple discharge of approximately 50 ml d-1. 3. PCB levels were high in adipose tissue (102 mg kg-1), serum, (512 micrograms l-1), skin lipids (66.3 mg kg-1), and in the nipple discharge (712 micrograms l-1). 4. The patient's history, the medical evaluation and prior unsuccessful symptomatic treatments were indicative of consequences elicited by occupational exposure to chemicals. 5. Detoxification treatment reduced the PCB levels in adipose tissue to 37.4 mg kg-1 and in serum to 261 micrograms l-1, a 63% and 49% reduction, respectively. 6. The nipple discharge ceased and the symptoms improved. 7. Excretion of intact PCBs in sebum was appreciable before treatment and was enhanced by up to five-fold during detoxification. 8. This therapeutic approach appears promising for cases involving occupational exposure to lipophilic chemicals.

Reduced adipsin mRNA and circulating adipsin protein are modulated by adrenal steroids in obese Zucker rats.

We investigated expression of the adipose-specific serine protease adipsin in genetically obese Zucker rats and whether adrenalectomy modifies expression. Adipsin mRNA levels were determined by slot blot and Northern blot analysis of total RNA samples extracted from epididymal adipose tissue and isolated retroperitoneal adipocytes of obese (fa/fa) and homozygous lean (Fa/Fa) Zucker rats. Both 30-day-old and 4-mo-old animals were analyzed in experiment 1. In experiment 2, 10-wk-old obese and lean rats were either bilaterally adrenalectomized or sham operated, and adipsin mRNA levels were determined on tissue and cell samples 2 wk postsurgery. In both experiments, serum adipsin protein was determined by Western blot analysis and plasma insulin by radioimmunoassay. The data show that both adipsin mRNA and adipsin protein are reduced in obese compared with lean rats and that adrenalectomy restores these values toward normal in obese rats. The data thus suggest that adrenal steroids are involved in modulating adipsin expression in obese Zucker rats and that insulin may be an intermediary factor in such modulation.

Essential fatty acid nutrition of the American alligator (Alligator mississippiensis).

The essential fatty acid (EFA) nutrition of young American alligators (Alligator mississippiensis) was examined by feeding a variety of fats/oils with potential EFA activity. Over a 12-wk period, alligators fed diets containing 2.5 or 5.0% chicken liver oil grew longer and heavier and converted feed to body mass more efficiently than alligators fed other fat/oil combinations that lacked or contained only trace amounts of arachidonic acid [20:4(n-6)]. Alligators fed an EFA-deficient diet (containing only coconut fat as the dietary fat) were the slowest-growing animals and converted feed to body mass least efficiently. However, over a 41-wk feeding period, alligators fed this diet showed no obvious external signs of deficiency other than being reduced in size and unthrifty. Fatty acid composition of heart, liver, muscle, skin and adipose tissue lipids was influenced markedly by dietary fat composition. Tissues varied significantly in response to dietary fat composition. Heart lipids contained the lowest levels of short- and medium-chain fatty acids and the highest levels of arachidonic acid. Arachidonic acid levels were less influenced by diet than were levels of other 20- and 22-carbon polyunsaturated fatty acids. Radiotracer studies indicated that linoleic acid was converted to arachidonic acid in the liver. Nevertheless, tissue arachidonic acid levels also appeared to be maintained by concentration from dietary sources and selective conservation. It appears that a dietary source of arachidonic acid may be required for a maximum rate of growth.

Utilization of ileal digestible amino acids by growing pigs: effect of dietary lysine concentration on efficiency of lysine retention.

Diets were formulated using sugar, soya-bean meal and free amino acids to contain 0.1-0.8 g lysine/MJ digestible energy (DE) and offered at three times maintenance to male and female pigs from 20 to 45 kg live weight. Growth responses and retentions of protein, fat, energy and lysine were assessed. Increasing the dietary lysine concentration resulted in significant (P less than 0.001) linear and curvilinear increases in growth rates and decreases in food conversion ratios. There was only a small effect of lysine concentration on total energy retention, but a substantial effect on the partitioning of energy deposition, with increases in the rate of protein deposition and decreases in fat retention. There was no difference in the efficiency of protein deposition between male and female pigs but males responded more to higher lysine concentrations than females (estimated 0.93 and 0.74 g lysine/MJ DE for males and females respectively). Lysine concentration in the protein deposited by the pigs increased linearly and curvilinearly (P less than 0.01) from 5.8 to 6.6 g lysine/16 g N with increasing dietary lysine concentration. There was a linear and quadratic response (P less than 0.001) in retention of ileal digestible lysine, with the minimum retention of 0.16 occurring at 0.1 g lysine/MJ DE and increasing to a maximum retention of 0.73 at a dietary concentration of 0.47 g lysine/MJ DE. The efficiency of lysine retained/ileal digestible lysine intake was 0.86 and the endogenous lysine loss was estimated at 0.94 g/d.

Relationships between adipose tissue characteristics of newborn pigs and subsequent performance: III. Histological and chemical characteristics of backfat.

Genetic and phenotypic relationships between adipose tissue characteristics of 253 piglets aged 8 d and qualitative characteristics of their backfat at 95 and 145 kg live weight were studied. Heritability of these qualitative aspects was estimated as well. Backfat samples were tested for cellularity, moisture, lipid content and fatty acid composition of the outer and inner layers. Adipose cellularity at birth proved to be correlated with cell volume, moisture and total unsaturated fatty acid, C18:0 and C18:2 contents present in backfat at slaughter. Estimates of heritability calculated for the outer layer of backfat indicated that progress can be made by selecting for water content and fatty acid composition. Neonatal cellularity furnishes a useful indicator not only of the backfat cell size at slaughter but also of the chemical factors important in determining the physical and organoleptic characteristics of porcine fat.

Norepinephrine turnover in brown adipose tissue of young rats: effects of rearing temperature.

Brown adipose tissue (BAT) has an important role in the thermogenesis of newborn mammals and cold-acclimated rodents. Heat production within the tissue is regulated by the sympathetic nervous system (SNS) via norepinephrine (NE) release. As only little is known of the progressive role of SNS during the developmental period, NE content and turnover were determined in BAT of infant rats 1-33 days of age. The dams and the pups were kept at either 28 or 16 degrees C. It was observed that the NE level in the tissue and the mediator turnover had an almost parallel evolution during the experimental period. In the first week of life, level and turnover of NE were higher in the 28 degrees C group than in the 16 degrees C group. During the second week, the level remained constant in rats kept at 28 degrees C, but the NE turnover slightly decreased. In contrast, a large increase of both parameters was observed in rats kept at 16 degrees C. Then a decrease occurred in both groups until weaning. Subsequently the values remained almost constant. It must be noted that, from the end of the first week, NE content and turnover were always significantly higher in the 16 degrees C group than in the 28 degrees C group. At the end of the first month, the level was 50% higher in cold-acclimated rats and turnover was four times as high as in controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Sex difference in inhalation toxicity of p-dichlorobenzene (p-DCB) in rats.

The organ distribution and toxicity of p-dichlorobenzene (p-DCB) were compared in male and female rats after inhalation of 500 ppm of p-DCB for 24 h in a whole-body chamber. Concentrations of p-DCB in the serum, liver, kidney and fatty tissues were measured by gas chromatography at intervals during and up to 24 h after the treatment. Though no significant differences in the serum levels were observed between male and female rats, the p-DCB values in the livers of female rats were significantly higher than those of male rats. Conversely, significantly higher levels were found in the kidneys of male than of female rats. The distribution results thus appeared to correlate with the fact that nephrotoxic changes were observed only in male rats and that the appearance of minor hepatotoxic changes was limited to females.

Body composition by photon absorptiometry.

Whole-body dual photon measurements of body composition were made in 9 controls, 15 athletes and 5 patients with eating disorders. Measurements of lean body mass, mineral mass and fat mass were accurate. Lean body mass and fat mass were measured with reliabilities of about 0.8 kg while for total body bone mineral mass, the reliability was about 60 g. Women had a smaller lean body mass and a greater fat mass than men. Women with eating disorders had the same body composition as males. Measurements of body composition by dual photon absorptiometry are safe and reliable.

Dual-energy x-ray absorptiometry for total-body and regional bone-mineral and soft-tissue composition.

Bone mineral density (BMD) and soft-tissue composition of the total body and major subregions were measured with dual-energy x-ray absorptiometry (DEXA). Total body scans were made in 12 young adults (6 male, 6 female) on five occasions at both a medium speed (20 min) and a fast speed (10 min). There were no significant differences in mean results or in precision errors between the two speeds. The precision errors (1 SD) for total body BMD, percent fat in soft tissue (% Fat), fat mass, and lean tissue mass were less than 0.01 g/cm2, 1.4%, 1.0 kg, and 0.8 kg, respectively. These results corresponded to a relative error of 0.8% for total body BMD and 1.5% for lean body mass. Regional BMD and soft-tissue values (arms, legs, trunk) were determined with slightly higher precision errors. Skeletal mineral was 5.8 +/- 0.5% of lean tissue mass (r = 0.96, p less than 0.001). DEXA provides precise composition analysis with a low radiation exposure (less than 0.1 microGy).

Determination of the alteration in fatty acid profiles, sensory characteristics and carcass traits of swine fed elevated levels of monounsaturated fats in the diet.

The present study was designed to determine the effects of supplemental fat or oil rich in oleic acid on the fatty acid profiles (FAP) and physical and sensory traits of pork carcasses. Sixty barrows and gilts were equally distributed among five dietary treatments consisting of a control diet of corn and soybean meal and four similar test diets that contained 10% animal fat (45.3 oleic), safflower oil (72.1 oleic), sunflower oil (80.9 oleic) or canola oil (57.7 oleic). The pigs were slaughtered after being fed these diets for 90 d at about 100 kg live weight. Carcass traits, FAP and sensory properties were evaluated for each treatment. First-rib fat thickness, ham muscling score and longissimus muscle areas were not different (P less than .05), but last-rib fat thickness was increased (P less than .05) with the supplemental dietary fat or oils. No differences existed for marbling scores, lean color, firmness or texture scores between the controls and pigs supplemented with either animal fat or safflower oil. However, pigs supplemented with sunflower or canola oil had lower marbling scores, lean color, firmness and texture scores. Fat became softer and more oily (P less than .05) with the supplemental dietary safflower, sunflower and canola oils. Sensory evaluation (loin chops) showed no differences (P less than .05) in sustained juiciness, tenderness or flavor intensity evaluations among treatments. However, the pigs fed canola oil had lower (P less than .05) flavor quality scores or overall palatability evaluations. Chops from the pigs fed canola oil also had 46% more off-flavors than all other treatments.(ABSTRACT TRUNCATED AT 250 WORDS)

Apolipoprotein E polymorphism alters the association between body fatness and plasma lipoproteins in women.

The aim of the present study was to investigate the associations between total adiposity, body fat distribution, and plasma lipoprotein levels within groups of women defined on the basis of apolipoprotein E phenotypes, in order to verify whether apoE polymorphism could modify these associations. In women having only apolipoprotein E3 isoforms (n = 24), body fat mass, the waist: hip circumference ratio, and computed tomography-derived total and intra-abdominal fat areas were all positively correlated with very low density lipoprotein (VLDL) and low density lipoprotein (LDL) lipids and apolipoprotein B concentrations. These body fatness variables were also negatively correlated with plasma high density lipoprotein (HDL) cholesterol concentration. These associations were, however, altered in the groups of women carrying either apoE2 or E4 isoforms. Indeed, in women carrying the apoE2 isoform (n = 22), body fatness variables were predominantly associated with VLDL components concentration (0.05 greater than P less than 0.01) and with LDL triglyceride content. No association was found between adiposity and LDL cholesterol or apolipoprotein B levels in these women. In contrast, no relationship was found between total adiposity, regional fat accumulation, and VLDL fraction in women carrying the apolipoprotein E4 isoform (n = 17). In this latter group, computed tomography-measured total abdominal fat accumulation was positively correlated with LDL apolipoprotein B (r = 0.58, P less than 0.05) concentration, whereas intra-abdominal fat accumulation was positively correlated with both LDL cholesterol and apolipoprotein B concentrations (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

A rapid and highly sensitive method for the quantitative determination of dexamethasone in plasma, synovial fluid and tissues by combined gas chromatography/negative ion chemical ionization mass spectrometry.

A new, simple and highly sensitive assay is developed for the quantitative measurement of very low levels of dexamethasone in human plasma, synovial fluid and tissues following a topical administration of the drug. Dexamethasone and the internal standard, flumethasone, are measured by gas chromatography/negative ion chemical ionization mass spectrometry with methane as the reagent gas. After a three-step extraction procedure, the two compounds of interest are converted to their trimethylsilyl ether derivatives using trimethylsilylimidazole and formamide as the base catalyst. Under soft derivatization conditions only one chromatographic peak corresponding to the trisubstituted derivative is observed. The mass spectrometer is focused to monitor abundant and stable characteristic high-mass ions (m/z 446 and 464) which are generated in the ion source by an electron capture process. This assay requires only 1 ml of plasma or 0.5 ml of synovial fluid and the detection limit of the method is equal to 0.1 ng ml-1 with a relative standard deviation lower than 6%.

Quantification of the alpha and beta subunits of the transducing elements (Gs and Gi) of adenylate cyclase in adipocyte membranes from lean and obese (ob/ob) mice.

The abundance of the alpha and beta subunits of the GTP-binding proteins (G-proteins) that transduce hormonal messages to adenylate cyclase was assessed in adipocyte membranes from lean (+/+) and obese (ob/ob) mice, using ADP-ribosylation with bacterial toxin and immunodetection. Both methods revealed two Gs alpha species (48 and 42 kDa) in the membranes. Compared with those of lean mice, the membranes from obese mice contained substantially less of the 48 kDa species of Gs alpha, as assessed by both methods. ADP-ribosylation by pertussis toxin showed that only half as much ADP-ribose was incorporated into Gi alpha in the membranes from obese as compared with lean mice. Immunodetection revealed two separate Gi alpha peptides (39 and 40 kDa) and showed that the 40 kDa species was less abundant in the membranes from obese mice, whereas the amount of the 39 kDa species was similar in membranes from both lean and obese animals. Based on ADP-ribosylation assays, in membranes from lean mice the ratio Gs alpha/Gi alpha was 1:16, whereas in the membranes from obese mice it was 1:10. Similar amounts of immunodetectable beta peptide were found in both types of membranes. On the basis of the currently accepted dissociation model of adenylate cyclase activation, the decrease in the abundance of the Gi alpha subunit in adipocyte membranes from obese mice could account for the abnormal kinetics of the enzyme in these membranes.