Home About us Contact
|
Home About us Contact | ||
|
|
||
Subcutaneous Adipose Tissue (subcutaneous + adipose_tissue)
Selected AbstractsIn humans the adiponectin receptor R2 is expressed predominantly in adipose tissue and linked to the adipose tissue expression of MMIF-1DIABETES OBESITY & METABOLISM, Issue 4 2010K. Kos In this study, the regional adipose tissue-adiponectin (AT-ADN) and adiponectin receptor (R1 and R2) expression and their relation with metabolic parameters, circulating and AT-derived cytokine expressions were compared. Paired subcutaneous adipose tissue (SCAT) and visceral adipose tissue (VAT) were taken from 18 lean and 39 obese humans, AT-mRNA expression of adipokines analysed by RT-PCR and corresponding serum levels by enzyme-linked immunosorbent assay (ELISA). R1 and R2 adipocyte expression was compared with 17 other human tissues. ADN-gene expression was lower in VAT than SCAT [mean (SD) 1.54 (1.1) vs. 2.84 (0.87); p < 0.001], and lower in obese subjects (VAT : p = 0.01;SCAT : p < 0.001). SCAT-ADN correlated positively with serum ADN (r = 0.33;p = 0.036) but not VAT-ADN. AT expressions of ADN and macrophage migration inhibiting factor (MMIF), IL18 and cluster of differentiation factor 14 (CD14) in both depots showed inverse correlations. R1 and R2 were expressed ubiquitously and R2 highest in SCAT, and this is much higher (×100) than R1 (×100). R expression was similar in lean and obese subjects and unrelated to the metabolic syndrome, however, receptors correlated with VAT-MMIF (R 1: r = 0.4;p = 0.008;R 2: r = 0.35,p = 0.02) and SCAT-MMIF expression (R 2: r = 0.43;p = 0.004). Unlike ADN, its receptors are expressed in many human tissues. Human R2 expression is not highest in the liver but in AT where it is associated with MMIF expression. The adiponectin-dependent insulin-sensitizing action of thiazolidinediones is thus probably to differ amongst species with weaker effects on the human liver. [source] Assessment of different techniques for subcutaneous glucose monitoring in Type 1 diabetic patients during ,real-life' glucose excursionsDIABETIC MEDICINE, Issue 3 2010J. K. Mader Diabet. Med. 27, 332,338 (2010) Abstract Aims, To compare the accuracy of two marketed subcutaneous glucose monitoring devices (Guardian RT, GRT; GlucoDay S, GDS) and standard microdialysis (CMA60; MD) in Type 1 diabetic patients. Methods, Seven male Type diabetic patients were investigated over a period of 26 h simulating real-life meal glucose excursions. Catheters of the three systems were inserted into subcutaneous adipose tissue of the abdominal region. For MD, interstitial fluid was sampled at 30- to 60-min intervals for offline glucose determination. Reference samples were taken at 15- to 60-min intervals. All three systems were prospectively calibrated to reference. Median differences, median absolute relative differences (MARD), median absolute differences (MAD), Bland,Altman plot and Clark Error Grid were used to determine accuracy. Results, Bland,Altman analysis indicated a mean glucose difference (2 standard deviations) between reference and interstitial glucose of ,10.5 (41.8) % for GRT, 20.2 (55.9) % for GDS and 6.5 (35.2) % for MD, respectively. Overall MAD (interquartile range) was 1.07 (0.39; 2.04) mmol/l for GRT, 1.59 (0.54; 3.08) mmol/l for GDS and 0.76 (0.26; 1.58) mmol/l for MD. Overall MARD was 15.0 (5.6; 23.4) % (GRT), 19.7 (6.1; 37.6) % (GDS) and 8.7 (4.1; 18.3) % (MD), respectively. Total sensor failure occurred in two subjects using GRT and one subject using GDS. Conclusions, The three investigated technologies had comparable performance. Whereas GRT underestimated actual blood glucose, GDS and MD overestimated blood glucose. Considerable deviations during daily life meal glucose excursions from reference glucose were observed for all three investigated technologies. Present technologies may require further improvement until individual data can lead to direct and automated generation of therapeutic advice in diabetes management. [source] Glucose-induced release of tumour necrosis factor-alpha from human placental and adipose tissues in gestational diabetes mellitusDIABETIC MEDICINE, Issue 11 2001M. T. Coughlan Abstract Aims, The cytokine tumour necrosis factor-alpha (TNF-,) has been implicated in the pathogenesis of insulin resistance in Type 2 diabetes mellitus, but limited data are available in relation to gestational diabetes mellitus (GDM), a disease in which similar biochemical abnormalities exist. We investigated the effect of exogenous glucose on the release of TNF-, from placental and adipose (omental and subcutaneous) tissue obtained from normal pregnant women, and women with GDM. Methods, Human tissue explants were incubated for up to 24 h and TNF-, concentration in the incubation medium quantified by ELISA. The effect of normal (5 mmol/l) and high (15 and 25 mmol/l) glucose concentrations on the release of TNF-, was assessed. Results, In placental and subcutaneous adipose tissues obtained from women with GDM (n = 6), TNF-, release was significantly greater under conditions of high glucose compared with normal glucose (placenta, 25 mmol/l 5915.7 ± 2579.6 and 15 mmol/l 4547.1 ± 2039.1 vs. 5 mmol/l 1897.1 ± 545.5; subcutaneous adipose tissue, 25 mmol/l 423.5 ± 207.0 and 15 mmol/l 278.5 ± 138.7 vs. 5 mmol/l 65.3 ± 28.5 pg/mg protein; P < 0.05). In contrast, there was no stimulatory effect of high glucose on TNF-, release by tissues obtained from normal pregnant women (n = 6) (placenta, 25 mmol/l 1542.1 ± 486.2 and 15 mmol/l 4263.3 ± 2737.7 vs. 5 mmol/l 5422.4 ± 1599.0; subcutaneous adipose tissue, 25 mmol/l 189.8 ± 120.4 and 15 mmol/l 124.5 ± 32.3 vs. 5 mmol/l 217.9 ± 103.5 pg/mg protein). Conclusions, These observations suggest that tissues from patients with GDM release greater amounts of TNF-, in response to high glucose. As TNF-, has been previously implicated in the regulation of glucose and lipid metabolism, and of insulin resistance, these data are consistent with the hypothesis that TNF-, may be involved in the pathogenesis and/or progression of GDM. Diabet. Med. 18, 921,927 (2001) [source] Quantitative model of cellulite: three-dimensional skin surface topography, biophysical characterization, and relationship to human perceptionINTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 4 2005L. K. Smalls Gynoid lipodystrophy (cellulite) is the irregular, dimpled skin surface of the thighs, abdomen, and buttocks in 85% of post-adolescent women. The distinctive surface morphology is believed to result when subcutaneous adipose tissue protrudes into the lower reticular dermis, thereby creating irregularities at the surface. The biomechanical properties of epidermal and dermal tissue may also influence severity. Cellulite-affected thigh sites were measured in 51 females with varying degrees of cellulite, in 11 non-cellulite controls, and in 10 male controls. A non-contact high-resolution three-dimensional (3D) laser surface scanner was used to quantify the skin surface morphology and determine specific roughness values. The scans were evaluated by experts and na,ve judges (n = 62). Body composition was evaluated via dual-energy X-ray absorptiometry; dermal thickness and the dermal,subcutaneous junction were evaluated via high-resolution 3D ultrasound and surface photography under compression. Biomechanical properties were also measured. The roughness parameters Svm (mean depth of the lowest valleys) and Sdr (ratio between the roughness surface area and the area of the xy plane) were highly correlated to the expert image grades and, therefore, designated as the quantitative measures of cellulite severity. The strength of the correlations among na,ve grades, expert grades, and roughness values confirmed that the data quantitatively evaluate the human perception of cellulite. Cellulite severity was correlated to BMI, thigh circumference, percent thigh fat, architecture of the dermal,subcutaneous border (ultrasound surface area, red-band SD from compressed images), compliance, and stiffness (negative correlation). Cellulite severity was predicted by the percent fat and the area of the dermal,subcutaneous border. The biomechanical properties did not significantly contribute to the prediction. Comparison of the parameters for females and males further suggests that percent thigh fat and surface area roughness deviation are the distinguishing features of cellulite. [source] Adipose tissue gene expression in obese dogs after weight lossJOURNAL OF ANIMAL PHYSIOLOGY AND NUTRITION, Issue 3 2008V. Leray Summary Body weight (BW) mainly depends on a balance between fat storage (lipogenesis) and fat mobilization (lipolysis) in adipocytes. BW changes play a role in insulin resistance (IR), the inability of insulin target tissue to respond to physiological levels of insulin. This results in inhibition of lipogenesis and stimulation of lipolysis. Weight gain leads to IR whereas, weight loss improves insulin sensitivity (IS). The aim of this study was to evaluate the effect of weight loss and recovery of IS on the expression of genes involved in lipogenesis and lipolysis in weight losing dogs. Gene expression was studied in both subcutaneous and visceral adipose tissue. Obese dogs received a hypoenergetic low fat high protein diet (0.6 × NRC recommendation). Before and after weight loss, IS was assessed using the euglycaemic hyperinsulinaemic clamp. Gene expression of IRS-2, SREBP, intracellular insulin effectors, ACC, FAS, FABP, ADRP, PEPCK, lipogenesis key proteins, perilipin and HSL, lipolysis key proteins were quantified using real-time RT-PCR in subcutaneous and visceral fat. BW decreased from 15.2 ± 0.5 to 11.4 ± 0.4 kg (p < 0.05) over 78 ± 8 days. When obese, dogs were insulin resistant. After weight loss, IS was improved. In the subcutaneous adipose tissue, the expression of only the IRS-2 was increased. In the visceral adipose tissue, the expression of the genes involved in the lipogenesis was decreased whereas one of the genes implied in the lipolysis did not change. The expression profile of genes involved in lipid metabolism, as measured after weight loss, is indicative for a lower lipogenesis after weight loss than in obese dogs. Our results also confirm dramatic differences in the lipid metabolism of visceral and subcutaneous fat. They should be completed by comparing gene expression during weight losing and normal weight steady state. [source] Minocycline hyperpigmentation isolated to the subcutaneous fatJOURNAL OF CUTANEOUS PATHOLOGY, Issue 7 2005Zakia Rahman We present a 15-year-old girl with bilateral lower extremity discoloration of one-year duration while taking minocycline for acne vulgaris. The clinical characteristics best supported type II minocycline hyperpigmentation, but the histology revealed that the pigmentation was solely limited to the subcutaneous adipose tissue, completely sparing the dermis. Special stain for iron was negative. This is the first case to our knowledge with pigment exclusively located in the subcutaneous fat and with the unusual finding of a negative stain for iron. [source] Merkel cell carcinoma: a clinicopathologic study with prognostic implicationsJOURNAL OF CUTANEOUS PATHOLOGY, Issue 3 2004Ryan T. Mott Background:, Merkel cell carcinoma (MCC) is a frequently aggressive neuroendocrine malignancy of the skin that presents in sun-exposed areas on elderly patients. Although originally described over 30 years ago, many aspects of MCC remain to be defined. Of particular importance is the need to identify prognostic factors capable of predicting the biological behavior of these tumors. Knowledge of these factors may help in determining which patients require more aggressive treatment regimens. In this study, we examined 25 cases of MCC with an attempt to identify clinical, histopathological, or immunohistochemical features capable of predicting disease outcome. Methods:, Features that we evaluated in each case included age, gender, race, tumor location, tumor size, depth of invasion, growth pattern, lymphocytic infiltration, mitotic activity, ulceration, necrosis, vascular invasion, and perineural invasion. In addition, we examined neural cell adhesion molecule and cytokeratin-20 expression using immunohistochemical methods. Results:, We found that most patients were males (84%) with an average age of 74 years. The tumors were located on the head and neck (68%) and upper extremities (32%). Overall, 64% of the patients developed metastatic disease to regional lymph nodes or distant sites (average follow-up time of 21 months). Local recurrence was also common, occurring in 29% of the patients. The overall 1- and 2-year survival rates were 80 and 53%, respectively. Histopathological examination revealed tumors with an average size of 7.2 mm. Common features included invasion into the subcutaneous adipose tissue, solid growth pattern, tumor necrosis, and vascular and perineural invasion. Findings that had a statistically significant correlation with poor outcome included tumor size ,5 mm (p = 0.047), invasion into the subcutaneous adipose tissue (p = 0.005), diffuse growth pattern (p = 0.040), and heavy lymphocytic infiltration (p = 0.017). The remaining findings, including the immunohistochemical results, did not correlate with disease outcome. Using logistic regression models, we show that depth of invasion and degree of lymphocytic infiltration are strong predictors of disease outcome. Conclusions:, The current controversies regarding the treatment of early-stage MCC (i.e., localized disease) underscore the importance of identifying clinicopathological features capable of predicting tumor behavior. In this study, we have identified several prognostic features in MCC. Perhaps, these features may prove useful in identifying patients who require more aggressive treatment regimens. [source] Threshold values of visceral fat and waist girth in Japanese obese childrenPEDIATRICS INTERNATIONAL, Issue 5 2005Kohtaro Asayama AbstractBackground:,In order to define the diagnostic criteria for visceral adipose tissue (VAT) accumulation and abdominal obesity in Japanese youths, a cross-sectional, multicenter study was conducted. Methods:,Subjects were 194 boys and 96 girls ranging in age from 6 to 15 years. Obese youths were classified according to the occurrence of abnormal values in serum triglyceride, alanine aminotransferase or insulin level. A threshold value of each criterion was calculated, using the analysis of receiver operating characteristic (ROC) curve. The areas of total abdominal adipose tissue (AT), VAT and subcutaneous adipose tissue (SAT) were estimated by single slice computed tomography at the level of umbilicus. Results:,VAT area was greater in boys than it was in girls. The critical values for VAT area and waist circumference in all subjects were 54.8 cm2 and 83.5 cm, respectively. The values for the area under the ROC curves were VAT area > total AT area > waist circumference > SAT area > percentage overweight > percentage body fat. The sensitivity and specificity for VAT area were 90.5 and 79.5%, respectively. Those for waist circumference were high enough (> 70%) for clinical use. In the linear regression analysis assigning VAT area as an independent variable and waist circumference as a dependent variable, the expected value for the waist circumference was 82 cm. Conclusion:,In Japanese obese youths ranging in age from 6 to 15 years, the diagnostic criteria for the waist circumference was 82 cm, and that for VAT area was 55 cm2. [source] Waist-to-hip ratio and adipose tissue distribution: Contribution of subcutaneous adiposityAMERICAN JOURNAL OF HUMAN BIOLOGY, Issue 3 2003Mark Daniel The waist-to-hip ratio (WHR) reflects the relative distribution of adipose tissue in the human body. However, whether this is due to the musculoskeletal structures of the waist and hip or the overlying subcutaneous adipose tissue has been disputed. We measured waist and hip girths in 11 male and 11 female cadavers, aged 55,94 years, before and after complete removal of skin and subcutaneous adipose tissue. Girths measured following removal of subcutaneous adipose tissue were termed "waist gx" and "hip gx", and their ratio "WHRx". Masses of regional adipose tissue segments were obtained by complete dissection, and the adipose mass ratios "trunk/arm-plus-leg", "trunk/leg", "internal/arm-plus-leg", and "internal/leg" were derived. As assessed by analysis of variance, WHR accounted for significant (P < 0.05) portions of the variance in all adipose mass ratios; adjustment for internal adipose mass increased the significance of all these relationships (P < 0.005). The ratio WHRx was not related to any ratio of regional adipose masses. Waist girth was related to trunk (P < 0.001) and internal (P < 0.05) adipose masses, and hip girth was related to arm-plus-leg adipose mass (P < 0.0001) and leg adipose mass (P < 0.0001), but waist gx and hip gx were not related to dependent variables. The results indicate that the ability of WHR and waist and hip girths to reflect the regional distribution of adipose tissue in the body is dependent upon the subcutaneous adipose tissue mass of the waist hip area, not its musculoskeletal constituency. Am. J. Hum. Biol. 15:428,432, 2003. © 2003 Wiley-Liss, Inc. [source] Breed difference and regulation of the porcine adipose triglyceride lipase and hormone sensitive lipase by TNF,ANIMAL GENETICS, Issue 6 2009T. Shan Summary Adipose triglyceride lipase (ATGL) and hormone sensitive lipase (HSL) are major novel triglyceride lipases in animals. The aim of this study was to determine if there are differences in the porcine ATGL (pATGL) and HSL genes between Jinhua pigs (a fatty breed) and Landrace pigs (a leaner breed). In addition, the effect of TNF, and pATGL-specific siRNA (pATGL-siRNA) on the expression of pATGL and HSL in porcine adipocytes was also examined. Compared with Landrace pigs, the body weight (BW) of Jinhua pigs was lower (P < 0.01), while intramuscular fat content (in the longissimus dorsi muscle), as well as the back fat thickness and body fat content were higher (P < 0.01). The expression of pATGL and HSL mRNA in Jinhua pigs was lower (P < 0.01) in subcutaneous adipose tissue, and greater (P < 0.01) in longissimus dorsi muscle compared with Landrace pigs. In vitro treatment of porcine adipocytes with TNF, decreased (P < 0.01) the glycerol release and the gene expression of pATGL, HSL and PPAR, in porcine adipocytes. Furthermore, transfection with pATGL-siRNA significantly decreased (P < 0.01) the expression of pATGL, while it had no effect on the expression of HSL. Treatment with 25 ng/ml TNF, in conjunction with pATGL-siRNA significantly decreased (P < 0.01) the expression of pATGL and HSL in cultured porcine adipocytes. These results provide useful information to further the understanding of the function of pATGL and HSL in porcine lipid metabolism, which should be applicable to the regulation of fat deposition and improvement of meat quality. [source] Slowed progression in models of huntington disease by adipose stem cell transplantation,ANNALS OF NEUROLOGY, Issue 5 2009Soon-Tae Lee MD Objective Adipose-derived stem cells (ASCs) are readily accessible and secrete multiple growth factors. Here, we show that ASC transplantation rescues the striatal pathology of Huntington disease (HD) models. Methods ASCs were isolated from human subcutaneous adipose tissue. In a quinolinic acid (QA)-induced rat model of striatal degeneration, human ASCs (1 million cells) were transplanted into the ipsilateral striatal border immediately after the QA injection. In 60-day-old R6/2 mice transgenic for HD, ASCs (0.5 million cells) were transplanted into each bilateral striata. In in vitro experiments, we treated mutant huntingtin gene-transfected cerebral neurons with ASC-conditioned media. Results In the QA model, human ASCs reduced apomorphine-induced rotation behavior, lesion volume, and striatal apoptosis. In R6/2 transgenic mice, transplantation of ASCs improved Rota-Rod performance and limb clasping, increased survival, attenuated the loss of striatal neurons, and reduced the huntingtin aggregates. ASC-transplanted R6/2 mice expressed elevated levels of peroxisome proliferator-activated receptor , coactivator-1, (PGC-1,) and reactive oxygen defense enzymes and showed activation of the Akt/cAMP-response element-binding proteins. ASC-conditioned media decreased the level of N-terminal fragments of mutant huntingtin and associated apoptosis, and increased PGC-1, expression. Interpretation Collectively, ASC transplantation slowed striatal degeneration and behavioral deterioration of HD models, possibly via secreted factors. Ann Neurol 2009;66:671,681 [source] Visceral adiposity is closely correlated with neck circumference and represents a significant indicator of insulin resistance in WHO grade III obesityCLINICAL ENDOCRINOLOGY, Issue 2 2010L. Yang Summary Objective, Although associations between visceral adiposity (intra-abdominal fat mass) and insulin resistance are well established, previous data include few subjects with WHO grade III obesity [body mass index (BMI) > 40 kg/m2]. We have investigated the relationship between visceral adiposity and insulin resistance using computed tomography (CT)-quantified fat mass and the homeostasis model assessment for insulin resistance (HOMA-IR) in patients with severe obesity. Patients and methods, Eighteen nondiabetic subjects with BMI > 40 kg/m2 were recruited. BMI, and waist, hip and neck circumferences were measured. Fasting plasma insulin and glucose were measured to calculate HOMA-IR. A single slice CT scan was taken at L4 and visceral and abdominal subcutaneous adipose tissue (VAT and ASAT, respectively) quantified using ,SliceOmatic' image analysis software. Results, A close correlation was demonstrated between VAT and HOMA-IR (r2 = 0·46, P = 0·002), whereas ASAT showed no relationship. Neck circumference correlated with both VAT (r2 = 0·67, P < 0·0001) and HOMA-IR (r2 = 0·35, P = 0·01). Waist circumference only correlated significantly with VAT (r2 = 0·25, P = 0·03). Conclusions, Visceral adiposity remains a strongly significant indicator of insulin resistance in WHO grade III obesity. Neck circumference surpasses other anthropometric measurements as a powerful marker of both VAT and insulin resistance. [source] Abnormal expression of PPAR gamma isoforms in the subcutaneous adipose tissue of patients with Cushing's diseaseCLINICAL ENDOCRINOLOGY, Issue 1 2007Fausto Bogazzi Summary Background, Obesity is a clinical feature of patients with Cushing's disease. Peroxisome proliferators-activated receptor (PPAR), is the master regulator of adipogenesis; however, the expression of PPAR, isoforms in the subcutaneous adipose tissue (SAT) of patients with Cushing's disease is unknown. Aim and methods, The expression of PPAR,1 and PPAR,2 was evaluated by real-time reverse transcription polymerase chain reaction (RT-PCR) and immunofluorescence (PPAR,2 only) in SAT samples of 7 patients with untreated active Cushing's disease (CushingUNTR), 8 with Cushing's disease in remission (CushingREM) after pituitary adenomectomy, 15 normal lean subjects (ControlLEAN), and 15 obese patients (ControlOBE). Results, ControlLEAN had a higher degree of PPAR,1 than PPAR,2 (PPAR,2/PPAR,1 ratio, 0·55 ± 0·35). PPAR,2/PPAR,1 ratio decreased in CushingUNTR (0·10 ± 0·043, P < 0·03 vs. ControlLEAN and ControlOBE), because of either increased PPAR,1 or reduced PPAR,2 expression. PPAR,2/PPAR,1 ratio was 0·48 ± 0·07 in CushingREM patients (P < 0·04 vs. CushingUNTR, P < 0·03 vs. ControlOBE). PPAR,2/PPAR,1 ratio was higher in ControlOBE 0·90 ± 0·38 than in ControlLEAN (P < 0·005 vs. ControlLEAN, P < 0·03 vs. CushingREM, P < 0·009 vs. CushingUNTR). PPAR,2/PPAR,1 ratio was related to serum cortisol levels only in patients with Cushing'disease (r = 0·688, P < 0·02). Conclusions, CushingUNTR patients had an abnormal expression of PPAR, isoforms in SAT related to serum cortisol levels. Although further studies are necessary, it is conceivable that variations in the expression of PPAR, isoforms might have a role in the abnormal adipogenesis of patients with Cushing's disease. [source] Lipid mobilization from human abdominal, subcutaneous adipose tissue is independent of sex during steady-state exerciseCLINICAL PHYSIOLOGY AND FUNCTIONAL IMAGING, Issue 4 2006Jens Bülow Summary The aim of the study was to elucidate whether there are sex differences of significant biological importance in the human abdominal, subcutaneous adipose tissue lipid metabolism when studied by Fick's Principle during rest and exercise in steady-state conditions. The net mobilization of fatty acids and glycerol from the abdominal, subcutaneous adipose tissue was measured by arterio-venous catheterizations and simultaneous measurements of adipose tissue blood flow with the local Xe-clearance technique in 16 healthy, young normal weight men and women during rest, during 1 h of exercise at moderate intensity, and for another 60 min during post-exercise recovery. The results show that there are not significant sex differences with respect to the steady-state fatty acid and glycerol mobilizations neither during resting condition nor during exercise. [source] Glucose-induced release of tumour necrosis factor-alpha from human placental and adipose tissues in gestational diabetes mellitusDIABETIC MEDICINE, Issue 11 2001M. T. Coughlan Abstract Aims, The cytokine tumour necrosis factor-alpha (TNF-,) has been implicated in the pathogenesis of insulin resistance in Type 2 diabetes mellitus, but limited data are available in relation to gestational diabetes mellitus (GDM), a disease in which similar biochemical abnormalities exist. We investigated the effect of exogenous glucose on the release of TNF-, from placental and adipose (omental and subcutaneous) tissue obtained from normal pregnant women, and women with GDM. Methods, Human tissue explants were incubated for up to 24 h and TNF-, concentration in the incubation medium quantified by ELISA. The effect of normal (5 mmol/l) and high (15 and 25 mmol/l) glucose concentrations on the release of TNF-, was assessed. Results, In placental and subcutaneous adipose tissues obtained from women with GDM (n = 6), TNF-, release was significantly greater under conditions of high glucose compared with normal glucose (placenta, 25 mmol/l 5915.7 ± 2579.6 and 15 mmol/l 4547.1 ± 2039.1 vs. 5 mmol/l 1897.1 ± 545.5; subcutaneous adipose tissue, 25 mmol/l 423.5 ± 207.0 and 15 mmol/l 278.5 ± 138.7 vs. 5 mmol/l 65.3 ± 28.5 pg/mg protein; P < 0.05). In contrast, there was no stimulatory effect of high glucose on TNF-, release by tissues obtained from normal pregnant women (n = 6) (placenta, 25 mmol/l 1542.1 ± 486.2 and 15 mmol/l 4263.3 ± 2737.7 vs. 5 mmol/l 5422.4 ± 1599.0; subcutaneous adipose tissue, 25 mmol/l 189.8 ± 120.4 and 15 mmol/l 124.5 ± 32.3 vs. 5 mmol/l 217.9 ± 103.5 pg/mg protein). Conclusions, These observations suggest that tissues from patients with GDM release greater amounts of TNF-, in response to high glucose. As TNF-, has been previously implicated in the regulation of glucose and lipid metabolism, and of insulin resistance, these data are consistent with the hypothesis that TNF-, may be involved in the pathogenesis and/or progression of GDM. Diabet. Med. 18, 921,927 (2001) [source] Quantitative analysis of human mitochondrial DNA using a real-time PCR assayHIV MEDICINE, Issue 3 2003K Gourlain Objectives Known for their ability to inhibit the human DNA polymerase-,, nucleoside analogues induce toxic effects on mitochondria ranging from increased serum lactate levels to fatal lactic acidosis. DNA polymerase-, ensures the mitochondrial DNA (mtDNA) replication and, thus, its inhibition leads to the decrease of the mtDNA. We describe a real-time PCR assay for mtDNA quantification associating DNA extraction procedures applied on peripheral blood mononuclear cells (PBMCs) and subcutaneous adipose tissues and to study the antiretroviral effect on mitochondria. Methods Total DNA was extracted from PBMCs and subcutaneous adipose tissues. Nuclear and mitochondrial genes were amplified to determine the number of copies of mtDNA per cell using a cyt-b recombinant plasmid as standard control. We analysed eight HIV-infected asymptomatic patients never treated, four patients who had been treated for 6 months with highly active antiretroviral therapy (HAART) and six non-infected donors. Results The mtDNA quantification gave rise to reproducible results as the mean coefficients of variation were 1.09% for replicates of samples undertaken 10 times within the same run, and 5.78% and 3.7% for replicates tested in five different runs at 1:100 and 1:1000 dilutions, respectively. Median levels of mtDNA in PBMCs of healthy donors, naive and treated HIV-infected patients were 2.94, 2.78 and 1.93 log HIV-1 RNA copies/mL, respectively. Whereas DNA from PBMCs was shown to be devoid of inhibitors, subcutaneous adipose tissues needed an extra treatment as they were found to be highly inhibited. Conclusions The method generated consistent and reproducible results and was successfully applied to DNAs extracted from PBMCs and subcutaneous adipose tissues with adapted extraction. The mtDNA changes in PBMCs were found to be fast as they fall off after 6 months' therapy, decreasing from 2.78 to 1.93 log copies/mL. [source] Hyperthermic injury to adipocyte cells by selective heating of subcutaneous fat with a novel radiofrequency device: Feasibility studiesLASERS IN SURGERY AND MEDICINE, Issue 5 2010Walfre Franco PhD Abstract Background and Objective The main objective of the present study is to demonstrate the feasibility of utilizing a novel non-invasive radiofrequency (RF) device to induce lethal thermal damage to subcutaneous adipose tissue only by establishing a controlled electric field that heats up fat preferentially. Study Design/Materials and Methods Adipocyte cells in six-well plates were subjected to hyperthermic conditions: 45, 50, 55, 60, and 65°C during 1, 2, and 3,minutes. Cell viability was assessed 72,hours after exposure. Two groups of abdominoplasty patients were treated with the RF device during and days before their surgical procedure. Temperatures of cutaneous and subcutaneous tissues were measured during treatment (3,minutes) of the first group. The immediate tissue response to heating was assessed by acute histology. The delayed tissue response was assessed by histology analysis of the second group, 4, 9, 10, 17, and 24 days after treatment (22,minutes). A mathematical model was used to estimate treatment temperatures of the second group. The model uses patient-based diagnostic measurements as input and was validated with in vivo clinical temperature measurements. Results Cell viability dropped from 89% to 20% when temperature increased from 45 to 50°C during 1,minute exposures. Three minutes at 45°C resulted in 40% viability. In vivo, the temperature of adipose tissue at 7,12,mm depth from the surface increased to 50°C while the temperature of cutaneous tissues was <30°C during RF exposure. Acute and longitudinal histology evaluations show normal epidermal and dermal layers. Subcutaneous tissues were also normal acutely. Subcutaneous vascular alterations, starting at day 4, and fat necrosis, starting at day 9, were consistently observed within 4.5,19,mm depth from the skin surface. Subcutaneous tissue temperatures were estimated to be 43,45°C for 15,minutes. Conclusions A controlled internal electric field perpendicular to the skin,fat interface is selective in heating up fat and, consequently, has the ability to induce lethal thermal damage to subcutaneous adipose tissues while sparing overlying and underlying tissues. In vitro adipocyte cells are heat sensitive to thermal exposures of 50 and 45°C on the order of minutes, 1 and 3,minutes, respectively. In vivo, 15,minutes thermal exposures to 43,45°C result in a delayed adipocyte cellular death response,in this study, 9 days. The novel RF device presented herein effectively delivers therapeutic thermal exposures to subcutaneous adipose tissues while protecting epidermal and dermal layers. Lasers Surg. Med. 42:361,370, 2010. © 2010 Wiley,Liss, Inc. [source] Organotypic culture of human bone marrow adipose tissuePATHOLOGY INTERNATIONAL, Issue 4 2010Kazuyoshi Uchihashi The precise role of bone marrow adipose tissue (BMAT) in the marrow remains unknown. The purpose of the present study was therefore to describe a novel method for studying BMAT using 3-D collagen gel culture of BMAT fragments, immunohistochemistry, ELISA and real-time reverse transcription,polymerase chain reaction. Mature adipocytes and CD45+ leukocytes were retained for >3 weeks. Bone marrow stromal cells (BMSC) including a small number of lipid-laden preadipocytes and CD44+/CD105+ mesenchymal stem cell (MSC)-like cells, developed from BMAT. Dexamethasone (10 µmol/L), but not insulin (20 mU/mL), significantly increased the number of preadipocytes. Dexamethasone and insulin also promoted leptin production and gene expression in BMAT. Adiponectin production by BMAT was <0.8 ng/mL under all culture conditions. Dexamethasone promoted adiponectin gene expression, while insulin inhibited it. This finding suggests that dexamethasone, but not insulin, may serve as a powerful adipogenic factor for BMAT, in which adiponectin protein secretion is normally very low, and that BMAT may exhibit a different phenotype from that of the visceral and subcutaneous adipose tissues. BMAT,osteoblast interactions were also examined, and it was found that osteoblasts inhibited the development of BMSC and reduced leptin production, while BMAT inhibited the growth and differentiation of osteoblasts. The present novel method proved to be useful for the study of BMAT biology. [source] | |||||||||||||||||||||||||