AIM: Comparison of manual and automatic (MagNA Pure) isolation methods of total RNA from adipose tissue with respect to its quality and recovery factor. MATERIAL: 120 human subcutaneous adipose tissue samples (about 100 mg/sample) were collected from patients during surgical operations. The tissue sample was stabilized in RNAlater (QIAGEN GmbH, Germany). METHODS: Total RNA was extracted by the following kits: Rneasy Protect Mini, Rneasy Lipid Tissue (QIAGEN GmbH, Germany) and MagNA Pure Compact RNA Isolation (Tissue) for MagNA Pure Compact Instrument (Roche Diagnostics GmbH, Germany). RESULTS: The average RNA yields with Rneasy Lipid Tissue kits were about two-fold higher in comparison with the Rneasy Protect Mini kit. When the MagNA Pure Compact System was used, RNA yields from the same sample were more uniform compared with manual systems. It was also more convenient and less time-consuming than the manual approach. No DNA contamination of total RNA samples was detected except for samples isolated by Rneasy Protect Mini Kit. CONCLUSION: Rneasy Lipid Tissue Kit and MagNA Pure Compact RNA Isolation Kit (Tissue) provide RNA samples of high quantity, purity and PCR amplificability. RNA samples are suitable for further processing using methods of molecular biology.

• MeSH
• DNA, Complementary genetics   MeSH
• Leptin genetics   MeSH
• Humans MeSH
• Molecular Biology methods   MeSH
• Reverse Transcriptase Polymerase Chain Reaction MeSH
• RNA isolation & purification   MeSH
• Adipose Tissue metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Comparative Study MeSH
• Names of Substances
• DNA, Complementary MeSH
• Leptin MeSH
• RNA MeSH

OBJECTIVE: To study the influence of chronic malnutrition in patients with anorexia nervosa on endocrine function of adipose tissue on both circulating and subcutaneous fat mRNA expression level. PATIENTS AND DESIGN: A total of 12 patients with anorexia nervosa and 18 normal weight age-matched women underwent anthropometric examination, single blood drawing and subcutaneous adipose tissue biopsy. MEASUREMENTS: Serum concentrations of high-sensitive CRP (hsCRP), leptin, soluble leptin receptor, adiponectin, resistin, interleukin-6 and insulin were measured by Luminex, enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) kits. Subcutaneous adipose tissue mRNA expression of the same adipokines, adiponectin receptors 1 and 2 and immunocompetent cells marker CD68 was measured by real-time polymerase chain reaction (PCR). RESULTS: Decreased body fat content of patients with anorexia nervosa was accompanied by reduced hsCRP, leptin and increased adiponectin and soluble leptin receptor. Resistin, interleukin-6 and insulin levels did not differ from those of the control group. Fat mRNA adiponectin, leptin, interleukin-6 and CD68 expression was reduced, resistin mRNA expression was increased and adiponectin receptor 1 and 2 expression were unchanged as compared to the control group. CONCLUSIONS: Local perturbations in resistin, adiponectin and interleukin-6 mRNA expression in subcutaneous adipose tissue are not reflected by its circulating levels. These changes could be involved in some local metabolic disturbances in subcutaneous adipose tissue of anorexia nervosa patients.

• MeSH
• Adiponectin blood   genetics   MeSH
• Antigens, Differentiation, Myelomonocytic genetics   MeSH
• C-Reactive Protein analysis   MeSH
• Antigens, CD genetics   MeSH
• Adult MeSH
• Gene Expression MeSH
• Adaptation, Physiological MeSH
• Interleukin-6 genetics   MeSH
• Insulin blood   MeSH
• Blood Glucose analysis   MeSH
• Receptors, Leptin blood   MeSH
• Humans MeSH
• Anorexia Nervosa immunology   metabolism   MeSH
• RNA, Messenger analysis   MeSH
• Statistics, Nonparametric MeSH
• Paracrine Communication * MeSH
• Subcutaneous Fat chemistry   immunology   metabolism   MeSH
• Malnutrition immunology   metabolism   MeSH
• Resistin blood   genetics   MeSH
• Case-Control Studies MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Comparative Study MeSH
• Names of Substances
• Adiponectin MeSH
• Antigens, Differentiation, Myelomonocytic MeSH
• C-Reactive Protein MeSH
• Antigens, CD MeSH
• CD68 antigen, human MeSH Browser
• Interleukin-6 MeSH
• Insulin MeSH
• Blood Glucose MeSH
• Receptors, Leptin MeSH
• RNA, Messenger MeSH
• Resistin MeSH

CONTEXT: Hyperglycemia and insulin resistance frequently occur in critically ill patients even without a history of diabetes. OBJECTIVE: Our objective was to study the role of adipose tissue hormonal production in the development of insulin resistance in cardiac surgery patients. PARTICIPANTS, INTERVENTIONS, AND SETTINGS: Fifteen patients with elective cardiac surgery underwent blood sampling before, at the end, and 6, 12, 24, 48, and 120 h after the end of their operation. Epicardial and sc adipose tissue sampling was done at the beginning and at the end of surgery in the Department of Cardiac Surgery. MAIN OUTCOME MEASURES: We measured serum concentrations and sc and epicardial adipose tissue mRNA expression of IL-6, monocyte chemoattractant protein-1 (MCP-1), TNF-alpha, leptin, resistin, and adiponectin and sc and epicardial adipose tissue mRNA expression of CD14, CD45, and CD68. RESULTS: The rate of insulin infusion required to maintain euglycemia increased up to 7-fold 12 h after the operation, suggesting the development of insulin resistance. Serum IL-6 levels increased 43-fold 12 h after surgery. MCP-1 peaked 6-fold at the end of surgery. Smaller peaks of TNF-alpha and leptin appeared 6 and 12 h after surgery, respectively. Resistin levels peaked 4-fold 24 h after surgery, but adiponectin levels were not significantly affected. TNF-alpha and CD45 mRNA expression increased markedly during the operation in sc adipose tissue. IL-6, resistin, and MCP-1 mRNA expression increased in both sc and epicardial adipose tissue. Leptin, adiponectin, CD14, and CD68 mRNA expression did not change significantly. CONCLUSIONS: Both sc and epicardial adipose tissue is a source of proinflammatory cytokines in cardiac surgery patients and may contribute to the development of postoperative insulin resistance.

• MeSH
• Anti-Inflammatory Agents blood   metabolism   MeSH
• Adipose Tissue, White metabolism   MeSH
• Biomarkers blood   MeSH
• Cytokines biosynthesis   physiology   MeSH
• Hormones blood   metabolism   MeSH
• Thoracic Surgery * MeSH
• Hyperglycemia blood   drug therapy   etiology   MeSH
• Immunocompetence physiology   MeSH
• Infusion Pumps MeSH
• Insulin administration & dosage   blood   MeSH
• Insulin Resistance physiology   MeSH
• Blood Glucose analysis   MeSH
• Middle Aged MeSH
• Humans MeSH
• Inflammation Mediators metabolism   physiology   MeSH
• RNA, Messenger metabolism   MeSH
• Pericardium cytology   MeSH
• Subcutaneous Fat metabolism   MeSH
• Postoperative Period MeSH
• Aged MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Clinical Trial MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Anti-Inflammatory Agents MeSH
• Biomarkers MeSH
• Cytokines MeSH
• Hormones MeSH
• Insulin MeSH
• Blood Glucose MeSH
• Inflammation Mediators MeSH
• RNA, Messenger MeSH