n-3 fatty acids and rosiglitazone improve insulin sensitivity through additive stimulatory effects on muscle glycogen synthesis in mice fed a high-fat diet.

AIMS/HYPOTHESIS: Fatty acids of marine origin, i.e. docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) act as hypolipidaemics, but they do not improve glycaemic control in obese and diabetic patients. Thiazolidinediones like rosiglitazone are specific activators of peroxisome proliferator-activated receptor gamma, which improve whole-body insulin sensitivity. We hypothesised that a combined treatment with a DHA and EPA concentrate (DHA/EPA) and rosiglitazone would correct, by complementary additive mechanisms, impairments of lipid and glucose homeostasis in obesity. METHODS: Male C57BL/6 mice were fed a corn oil-based high-fat diet. The effects of DHA/EPA (replacing 15% dietary lipids), rosiglitazone (10 mg/kg diet) or a combination of both on body weight, adiposity, metabolic markers and adiponectin in plasma, as well as on liver and muscle gene expression and metabolism were analysed. Euglycaemic-hyperinsulinaemic clamps were used to characterise the changes in insulin sensitivity. The effects of the treatments were also analysed in dietary obese mice with impaired glucose tolerance (IGT). RESULTS: DHA/EPA and rosiglitazone exerted additive effects in prevention of obesity, adipocyte hypertrophy, low-grade adipose tissue inflammation, dyslipidaemia and insulin resistance, while inducing adiponectin, suppressing hepatic lipogenesis and decreasing muscle ceramide concentration. The improvement in glucose tolerance reflected a synergistic stimulatory effect of the combined treatment on muscle glycogen synthesis and its sensitivity to insulin. The combination treatment also reversed dietary obesity, dyslipidaemia and IGT. CONCLUSIONS/INTERPRETATION: DHA/EPA and rosiglitazone can be used as complementary therapies to counteract dyslipidaemia and insulin resistance. The combination treatment may reduce dose requirements and hence the incidence of adverse side effects of thiazolidinedione therapy.

Low-dose acetylsalicylic acid inhibits the secretion of interleukin-6 from white adipose tissue.

BACKGROUND: Chronically elevated interleukin-6 (IL-6) is implicated in obesity-associated pathologies, where a proportion of this cytokine is derived from adipose tissue. Proinflammatory prostaglandins, which regulate this cytokine elsewhere, are also produced by this tissue. OBJECTIVE: To investigate whether constitutively active cyclooxygenase (COX)/prostaglandin (PG) pathway in white adipose tissue (WAT) is responsible for basal IL-6 production. DESIGN: The effect of acetylsalicylic acid (ASA), an inhibitor of COX, on IL-6 was assessed in human subjects and mice. COX, downstream PG synthase (PGS) activity and PG receptor signalling were determined in subcutaneous (SC), gonadal (GN) WAT and adipocytes. METHODS AND RESULTS: In obese humans, low-dose ASA (150 mg day(-1) for 10 days) inhibited systemic IL-6 and reduced IL-6 release from SC WAT ex vivo (0.2 mM). Similarly, in mice, ASA (0.2 and 2.0 mg kg(-1)) suppressed SC WAT 6-keto-PGF(1alpha) (a stable metabolite of prostacyclin) and IL-6 release. Although both COX isoforms are comparably expressed, prostacyclin synthase expression is higher in GN WAT, with levels of activity correlating directly with IL-6. Both ASA (5 mM) and NS-398 (COX-2 selective inhibitor

Orthosilicic acid stimulates collagen type 1 synthesis and osteoblastic differentiation in human osteoblast-like cells in vitro.

Silicon deficiency in animals leads to bone defects. This element may therefore play an important role in bone metabolism. Silicon is absorbed from the diet as orthosilicic acid and concentrations in plasma are 5-20 microM. The in vitro effects of orthosilicic acid (0-50 microM) on collagen type 1 synthesis was investigated using the human osteosarcoma cell line (MG-63), primary osteoblast-like cells derived from human bone marrow stromal cells, and an immortalized human early osteoblastic cell line (HCC1). Collagen type 1 mRNA expression and prolyl hydroxylase activity were also determined in the MG-63 cells. Alkaline phosphatase and osteocalcin (osteoblastic differentiation) were assessed both at the protein and the mRNA level in MG-63 cells treated with orthosilicic acid. Collagen type 1 synthesis increased in all treated cells at orthosilicic acid concentrations of 10 and 20 microM, although the effects were more marked in the clonal cell lines (MG-63, HCCl 1.75- and 1.8-fold, respectively, P < 0.001, compared to 1.45-fold in the primary cell lines). Treatment at 50 microM resulted in a smaller increase in collagen type 1 synthesis (MG-63 1.45-fold, P = 0.004). The effect of orthosilicic acid was abolished in the presence of prolyl hydroxylase inhibitors. No change in collagen type 1 mRNA level was seen in treated MG-63 cells. Alkaline phosphatase activity and osteocalcin were significantly increased (1.5, 1.2-fold at concentrations of 10 and 20 microM, respectively, P < 0.05). Gene expression of alkaline phosphatase and osteocalcin also increased significantly following treatment. In conclusion, orthosilicic acid at physiological concentrations stimulates collagen type 1 synthesis in human osteoblast-like cells and enhances osteoblastic differentiation.

Dexamethasone and retinoic acid differentially regulate growth and differentiation in an immortalised human clonal bone marrow stromal cell line with osteoblastic characteristics.

Clonogenic immortalised human pre-osteoblastic cell lines provide useful species-specific experimental tools for the study of the regulation of osteoblastic proliferation and differentiation. Steroid hormones are major regulators of bone formation. Although much is known about the effects of dexamethasone on osteoblastic growth and differentiation in vitro, there is less information on the effects of trans-retinoic acid (RA), particularly in human cultures. We have established a clonal adult human cell line (C1) derived from a bone marrow aspirate. The cell line appeared to be bi-potential. The cells were able to differentiate into an adipocytic phenotype under appropriate culture conditions. When grown in osteogenic medium, the cells expressed alkaline phosphatase (ALP) and osteocalcin mRNA. The C1 cells also expressed several other osteoblastic markers such as collagen type 1 (COL 1), PTH/PTH-rp receptor constitutively. Transcripts for the osteoblast transcription factor Cbfa1 was also detected under basal conditions. In addition treatment with 1,25(OH)(2)D(3) (10(-7)M) led to a marked increase in osteocalcin mRNA expression suggesting that this cell line represents a pre-osteoblastic population. We compared the effects of Dex and RA on osteoblastic function. For the assessment of PTH/PTH-rp receptor, osteocalcin and Cbfa1 mRNA expression and PTH-stimulated adenylate cyclase responsiveness, the cells were grown in the presence of Dex and RA and harvested on Days 1, 3, 7 and 14. RA (10(-7)M) had a mitogenic effect on the C1 cells. In contrast, Dex (10(-7)M) inhibited proliferation. A similar effect was observed with primary human bone marrow stromal cultures. Both Dex and RA inhibited COL 1 synthesis and decreased COL1 mRNA. Dex stimulated ALP activity and increased ALP mRNA expression whilst RA had an inhibitory effect. Dex treatment led to an increase in PTH/PTH-rp receptor mRNA and PTH-induced cAMP accumulation with a peak response at 24 h and this effect was sustained for up to 14 days. In contrast, long-term culture with RA resulted in a reduction in the cAMP response to PTH (Days 7 and 14) with no effect on PTH/PTH-rp receptor mRNA expression. Osteocalcin and Cbfa1 mRNA expression did not alter in the presence of Dex and RA at these time points. This study shows that Dex and RA have differential effects on the expression of the phenotypic markers and genes associated with osteoblast maturation. This homogeneous cell line can therefore be used further to elucidate the cellular and molecular mechanisms of action of Dex and RA at the different developmental stages of human osteoblastic differentiation. This cell line may thus provide a useful species-specific in vitro model for the evaluation of key genes and signalling molecules involved in osteogenesis. This would be of help in the design of 'in vivo' studies.

Computerized tomography of the distal radioulnar joint: correlation with ligamentous pathology in a cadaveric model.

The kinematics of the normal distal radioulnar joint (DRUJ) and the stabilizing function of various structures about the DRUJ were investigated in a study involving six fresh frozen cadavers. Sequential division of the supporting structures was correlated with abnormalities detected by computerized tomography (CT). The infratendinous portion of the extensor carpi ulnaris is a major restraint against dorsal and palmar subluxation. Division of the radioulnar ligaments and triangular fibrocartilage alone produced only minor changes. Lateral displacement was controlled by the interosseous membrane and the pronator quadratus. CT is a useful method of gaining objective and quantifiable information regarding incongruity of the DRUJ. Three scans--one each in pronation, neutral, and supination--would be optimal. The pronation scan is likely to detect palmar subluxation, while the neutral scan is sensitive for dorsal subluxation and DRUJ diastasis. The supination view should confirm reduction of any subluxation. An important observation of this study was the spontaneous reduction of palmar, dorsal, and lateral displacement in supination. This suggests that immobilization in supination may be indicated in cases of acute DRUJ injury.

Centrode characteristics of the lumbar spine as a function of segmental instability.

A cadaveric study provides a quantitative measurement of early degenerative disc disease before significant radiographic changes are present. Accurate assessment of spinal motion is hampered by the inaccessibility of the lumbar joints and because of the small range of motion at each motion segment. Using Moiré fringes and, more recently, computerized digitization of spinal motion, the authors have developed a technique of measuring centers of rotation for small ranges of motion. The centers of rotation when joined form a locus that has characteristics that allow one to identify spines with degenerative disc disease. These loci, or centrodes, are longest in the earliest stages of degeneration, but maintain their length through moderate degenerative disc disease. Furthermore, radiographic changes consistent with moderate disc disease are associated with inferior migration of the centrode.

Centrode patterns in the lumbar spine. Baseline studies in normal subjects.

Segmental spinal instability is a known cause of back pain, but no method of accurately quantifying instability exists. The movement of complex joints with rotational and translational components (such as the lumbar motion segment) is tracked by a pathway of instantaneous centers of rotation, or a centrode. Instability, ie, excessive and/or erratic movement, is reflected by increased centrode length in cadaver studies. This study describes an in vivo method that precisely determines the centrode pattern and reports the results for 21 normal male volunteers who were studied at the L4-5 and L5-S1 levels. Each volunteer underwent lateral radiographs of the lumbar spine. High speed films were used and six positions, from full extension to full flexion, were recorded. Acetate tracings and contour matching techniques recorded the relative positions of the vertebral bodies on each film. Multiple tracings of each radiograph, combined with a digitizer and computer, were used to improve precision in the calculated centrode patterns. Centrode lengths measured 43.7 mm and 55.9 mm, respectively, for the L4-5 and L5-S1 levels. This study demonstrates that precise centrode pattern analysis for sagittal plane motion of the lumbar spine is possible in vivo. Studies are under way to determine whether this technique will be useful as a clinical test in diagnosing early segmental instability of the lumbar spine in patients with low-back pain.