C-type natriuretic peptide stimulates osteoblastic proliferation and collagen-X expression but suppresses fibroblast growth factor-23 expression in vitro.

BACKGROUND: The effects of C-type natriuretic peptide (CNP) and fibroblast growth factor (FGF)-23 appear to oppose each other during the process of bone formation, whereas few studies exist on the interaction between CNP and FGF-23. The main objective of the present study is to probe whether CNP is directly responsible for the regulation of osteoblast or via antagonizing FGF-23. METHODS: Osteoblasts were cultured in the absence or presence of CNP (0, 10, and 100 pmol/L) for 24 h, 48 h and 72 h, respectively. RESULTS: The findings of the present study indicated that: (1) CNP significantly stimulated osteoblastic proliferation and collagen (Col)-X expression; (2) both osteoblastic (osteocalcin, procollagen type I carboxy-terminal propeptide, total alkaline phosphatase and bone-specific alkaline phosphatase) and osteolytic (tartrate-resistant acid phosphatase and cross-linked carboxyterminal telopeptide of type I collagen) bone turnover biomarkers were up-regulated by CNP in osteoblasts; (3) FGF-23 mRNA and protein were significantly down-regulated at 24 h by CNP in osteoblasts, but the expression of FGF receptor-1/Klotho had no significant change. CONCLUSIONS: CNP stimulates osteoblastic proliferation and Col-X expression via the down-regulation of FGF-23 possibly in vitro. However, the specific mechanisms of the interaction between CNP and FGF-23 in osteoblasts are still unclear according to our findings. A further study on osteoblasts cultured with CNP and FGF-23 inhibitor will be undertaken in our laboratory.

17ß-Estradiol improves osteoblastic cell function through the Sirt1/NF-κB/MMP-8 pathway.

Objective: This study aims to investigate the beneficial effects of 17ß-estradiol supplementation on the function of osteoblastic cells through the Sirtuin-1/nuclear transcription factor-κB/matrix metalloproteinase-8 (Sirt1/NF-κB/MMP-8) pathway.Methods: Mouse primary osteoblasts were obtained from neonatal mouse calvaria, and the cells were treated with or without 17ß-estradiol. We first detected the effect of 17ß-estradiol on the function of osteoblastic cells. Then, the changes in estrogen receptor-α (ERα), Sirt1, NF-κB, and MMP-8 were determined after the osteoblasts were treated with 17ß-estradiol. During supplementation with 17ß-estradiol, knockdown of Sirt1 in osteoblasts was used to further measure the changes of NF-κB and MMP-8 and observe the cell function.Results: In primary osteoblastic cells, exposure to 17ß-estradiol improved cell viability and increased the levels of bone formation biomarkers, including osteocalcin, osteoprotegerin (OPG), procollagen type 1 N-terminal propeptide (P1NP), and alkaline phosphatase (ALP). In addition, 17ß-estradiol supplement activated ERα and Sirt1 expression and inhibited NF-κB and MMP-8 expression. Moreover, these effects induced by 17ß-estradiol were reversed by knockdown of Sirt1 in mouse primary osteoblasts.Conclusion: 17ß-Estradiol replacement therapy may treat postmenopausal osteoporosis by improving osteoblastic cell function via the Sirt1/NF-κB/MMP-8 pathway.

Separate and Combined Effects of GIP and GLP-1 Infusions on Bone Metabolism in Overweight Men Without Diabetes.

CONTEXT: The gut-derived incretin hormones glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide 1 (GLP-1) have been suggested to play a role in bone metabolism. Exogenous administration of GIP inhibits bone resorption, but the effect of GLP-1 is less clear. Furthermore, the combined effect of exogenous GIP and GLP-1 on bone metabolism is unknown. OBJECTIVE: To investigate the effect of separate and combined infusions of the incretin hormones GIP and GLP-1 on bone resorption and formation. DESIGN: Randomized, double-blinded, placebo-controlled, crossover study including five study days. PARTICIPANTS: Seventeen overweight/obese men. INTERVENTIONS: On the first study day, a 50-g oral glucose tolerance test (OGTT) was performed. On the next four study days, isoglycemic IV glucose infusions (IIGI), mimicking the glucose excursions from the OGTT, were performed with concomitant infusions of GIP (4 pmol/kg/min), GLP-1 (1 pmol/kg/min), GIP+GLP-1 (4 and 1 pmol/kg/min, respectively), or placebo, respectively. PRIMARY OUTCOMES: Changes in bone resorption assessed by measurements of carboxy-terminal type I collagen crosslinks (CTX) and in bone formation as assessed by procollagen type 1 N-terminal propeptide (P1NP) concentrations. RESULTS: During the OGTT, CTX was significantly lowered by 54 ± 13% from baseline (mean ± SD) compared with 28 ± 12% during IIGI + saline (P < 0.0001). During IIGI+GLP-1 and IIGI+GIP, CTX was lowered by 65 ± 16% and 74 ± 9%, respectively, from baseline, whereas IGII+GIP+GLP-1 lowered CTX by 84 ± 4% from baseline. P1NP levels were unaffected by the interventions. CONCLUSIONS: Our data suggest that GLP-1, like GIP, may be involved in regulation of bone resorption and that GIP and GLP-1 together have partially additive inhibitory effects.

The medicinal mushroom Auricularia auricula-judae (Bull.) extract has antioxidant activity and promotes procollagen biosynthesis in HaCaT cells.

In this study, Auricularia auricula-judae (Bull.) extract (AAE) had potent antioxidant activity in vitro and promoted the biosynthesis of procollagen, a precursor of collagen in HaCaT cells. In addition, the expression of HAS-3 (hyaluronic acid synthase), which is a moisturizing factor, was increased in HaCaT cells in response to AAE. Therefore, this work suggests that AAE has the potential to exhibit antioxidant activity and promote procollagen biosynthesis in HaCaT cells.

Does metformin treatment influence bone formation in patients with nonalcoholic fatty liver disease?

Antidiabetic drug metformin that improves insulin sensitivity and used in the treatment of nonalcoholic fatty liver disease (NAFLD), may affect the bone health. Our study was designed to investigate a possible effect of metformin on bone formation marker, procollagen type I N-terminal propeptide (P1NP) in patients with NAFLD.In a randomized, placebo controlled study, 63 patients with NAFLD were assigned to one of 2 groups: Group 1 received daily metformin and Group 2 received placebo. Metabolic parameters, insulin resistance markers, and P1NP were determined.Although circulating P1NP levels did not differ significantly between the groups at baseline, at the end of the study, P1NP was significantly lower in patients treated with metformin than in the placebo group (p<0.007). Within-group analysis indicated that P1NP levels significantly decreased (p=0.023) in patients receiving metformin during 4-month follow-up period, while no change in P1NP was observed in placebo group (p=0.359). In general linear model metformin treatment was the only significant independent predictor of endpoint P1NP.Metformin treatment was associated with decrease in P1NP levels in patients with NAFLD. The effect on P1NP was independent of glucose lowering effect and caused from exposure to metformin per se.

Biochemical markers of bone turnover in diabetes patients--a meta-analysis, and a methodological study on the effects of glucose on bone markers.

UNLABELLED: This study examined whether markers of bone turnover differ between individuals with and without diabetes. Bone markers showed heterogeneity between studies and were discrepant for markers of bone creation and markers of bone degradation. Bone markers may be of lesser value in diabetes due to heterogeneity. INTRODUCTION: The aim of this meta-analysis was to compare existing literature regarding changes in bone markers among diabetics compared to healthy controls. To exclude that blood glucose levels among diabetes patients could influence the assays used for determining bone turnover markers, a methodological study was performed. METHODS: Medline at Pubmed Embase, Cinahl, Svemed+, Cochrane library, and Bibliotek.dk was searched in August 2012. The studies should examine biochemical bone turnover among diabetes patients in comparison to controls in an observational design. In the methodological study, fasting blood samples were drawn from two individuals. Glucose was added to the blood samples in different concentrations and OC, CTX, and procollagen type 1 amino terminal propeptide were measured after 0, 1, 2, and 3 h. RESULTS: Twenty-two papers fulfilled the criteria for the meta-analysis. From the pooled data in the meta-analysis, the bone markers osteocalcin (OC) (-1.15 ng/ml [-1.78,-0.52]) and C-terminal cross-linked telopeptide (CTX) (-0.14 ng/ml [-0.22, -0.05]) were significantly lower among diabetes patients than non-diabetes patients, however other markers did not differ. All markers displayed very high heterogeneity by I2 statistics. In the methodological study, the addition of glucose did not significantly change the bone markers neither by level of glucose nor with increasing incubation time. CONCLUSION: The dissociative pattern of biochemical bone markers of bone formation and bone resorption present in diabetes patients is thus not caused by glucose per se but may be modulated by unknown factors associated with diabetes mellitus.

Biomarker and imaging responses to spironolactone in subclinical diabetic cardiomyopathy.

BACKGROUND: Subclinical diabetic cardiomyopathy (DCM) is frequent in asymptomatic subjects with type 2 diabetes (T2DM). We sought the response of functional and fibrosis markers to therapy in a trial of aldosterone antagonism for treatment of DCM. METHODS: Biochemical, anthropometric, and echocardiographic data were measured in 225 subjects with T2DM. Myocardial function was evaluated with standard echocardiography and myocardial deformation; ischaemia was excluded by exercise echocardiography. Calibrated integrated backscatter and post-contrast T1 mapping from cardiac magnetic resonance imaging were used to assess myocardial structure. Amino-terminal propeptides of pro-collagen type I (PINP) and III (PIIINP), the carboxy-terminal propeptide of pro-collagen type I (PICP) and transforming growth factor beta-1 were measured from peripheral blood or urine to assess myocardial collagen turnover. RESULTS: Diastolic dysfunction was identified in 81 individuals, of whom 49 (25 male, age 60 ± 10 years) were randomized to spironolactone 25 mg/day or placebo therapy for 6 months. Groups were well-matched at baseline. Spironolactone therapy was associated with improvements in diastolic filling profile (Δpeak E wave velocity -4 ± 15 vs. 9 ± 10 ms, P = 0.001; ΔE/A ratio -0.1 ± 0.3 vs. 0.2 ± 0.2, P < 0.001) and cIB values (-21.2 ± 4.5 dB vs. -18.0 ± 5.2 dB, P = 0.026; ΔcIB -5.1 ± 6.8 vs. -1.3 ± 5.2, P = 0.030). ΔcIB was independently associated with spironolactone therapy (ß = 0.320, P = 0.026) but not Δblood pressure. With intervention, pro-collagen biomarkers (ΔPINP P = 0.92, ΔPICP P = 0.25, ΔPIIINP P = 0.52, and ΔTGF-ß1 P = 0.71) and T1 values (P = 0.54) remained similar between groups. CONCLUSIONS: Spironolactone-induced changes in myocardial structure and diastolic properties in DCM are small, and are unassociated with changes in collagen biomarkers or T1 values.

Antifibrotic effect of pirfenidone on human pterygium fibroblasts.

OBJECTIVE: The effects of pirfenidone were investigated on cultured human pterygium fibroblasts (HPFs). METHODS: HPFs were obtained from pterygium surgery and subjected to primary culture. After treatment with 0.5, 1.0 or 1.5 mg/mL pirfenidone, MTT and cell migration assays were performed, and procollagen secretion and TGF-ß expression were measured by Western blotting and immunofluorescence analysis. RESULTS: Pirfenidone had a significant inhibitory effect on HPF proliferation, migration and collagen synthesis. There were no differences between the cells treated with 0.5, 1.0 and 1.5 mg/mL pirfenidone and the controls in the MTT assay. After 48 h of treatment with 1.0 or 1.5 mg/mL pirfenidone, TGF-ß expression was significantly decreased. CONCLUSIONS: These findings demonstrate that pirfenidone inhibits the proliferation, migration and procollagen secretion of HPFs at nontoxic concentrations by decreasing TGF-ß expression. Thus, pirfenidone may be considered as a safe adjuvant for pterygium surgery to prevent recurrence.

Sensitivity of human dental pulp cells to eighteen chemical agents used for endodontic treatments in dentistry.

To determine the adverse effects against human dental pulp tissue, the sensitivity of human dental pulp cells (D824 cells) to 18 chemical agents used for endodontic treatments in dentistry was examined. The cytotoxicity, as determined by a decrease in colony-forming ability of cells treated with the chemical agents, increased as the concentration increased. As a quantitative measure of the cytotoxic effect, LC(50), the concentration which induces a 50% lethality, was extrapolated from the concentration-response curves. The rank of the chemical agents according to their cytotoxic effect (LC(50)) was sodium arsenite > formaldehyde > hydrogen peroxide > zinc oxide > thymol ≈ iodoform ≈ eugenol > guaiacol > ethylenediaminetetraacetic acid ≈ iodine > procaine > lidocaine ≈ chloramphenicol ≈ m-cresol > calcium hydroxide ≈ sodium hypochlorite ≈ phenol ≈ p-phenolsulfonic acid. To compare the cytotoxicity and the levels of apoptosis and mRNA expression of five genes related to the function of dental pulp tissue, D824 cells treated with the LC(50) concentrations of chemical agents were assayed by the TUNEL method and quantitative reverse transcription polymerase chain reaction analysis, respectively. The inducibility of apoptotic cells and the level of mRNA expression of the genes varied with the chemical agents, indicating that both effects occurred independent of the rank of cytotoxic effect of the chemical agents. The results not only provide information concerning cytotoxicity of various chemical agents to human dental pulp cells, but also show an insight into the diversity of the pharmacodynamic action of the chemical agents.

The potential effect of botulinum toxin type A on human dermal fibroblasts: an in vitro study.

BACKGROUND: The use of botulinum toxin type A (BoNT) continues to expand. Some physicians have noted a face-lifting effect after intradermal injection of BoNT, although the effects are controversial. OBJECTIVE: To investigate the in vitro effects of BoNT on human dermal fibroblasts. METHODS: The proliferation and toxic effects of BoNT on human dermal fibroblasts were measured. To understand the mechanism of BoNT on collagen production of fibroblasts, procollagen type I carboxy-terminal peptide (PIP) was measured using enzyme-linked immunosorbent assay, and collagen production was monitored using Western blotting. To examine the effect of BoNT on collagen degradation, we evaluated matrix metalloproteinase (MMP) production using gelatin zymography. RESULTS: BoNT did not stimulate the proliferation of or show toxic effects on human dermal fibroblasts. Levels of PIP increased significantly in fibroblasts grown in the presence of BoNT, and BoNT upregulated the expression of type I collagen and decreased the production of some MMPs in fibroblasts that prevent collagen degradation. CONCLUSIONS: This study shows interesting effects of BoNT on collagen production and degradation of human dermal fibroblasts in vitro. This research provides the experimental background for using intradermal BoNT injection for remodeling of dermal tissues in aged skin.

Effects of 1-year administration of olmesartan on portal pressure and TGF-beta1 in selected patients with cirrhosis: a randomized controlled trial.

BACKGROUND: The renin-angiotensin system plays an important role in hepatic fibrosis and portal hypertension. We evaluated the long-term effects of olmesartan, an angiotensin type 1 (AT1) receptor blocker, on hemodynamics and liver fibrosis. METHODS: Forty-eight selected patients with cirrhosis were randomly divided into two groups of 24 patients each, those who received and those who did not receive olmesartan treatment for 1 year. Hepatic hemodynamic studies, and measurements of transforming growth factor-beta1 (TGF-beta1) and blood markers of hepatic fibrosis, including serum hyaluronic acid (HA), type IV collagen, and procollagen III N-terminal propeptide levels, were also performed at the beginning and end of the study. RESULTS: The median dose of the final drug administration was 20 mg (range 10-40 mg). Olmesartan reduced the hepatic venous pressure gradient (HVPG) by -12.9 ± 9.1% (p = 0.035) after 1 year. No significant changes were seen in controls. Six of the 24 patients (25%) in the olmesartan group showed a >20% reduction of HVPG from baseline values. TGF-beta1 was significantly decreased in patients who received olmesartan (7.0 ± 8.2 vs. 3.1 ± 1.6 ng/mL, p = 0.046) but there was no decrease in the controls. A significant trend was shown by correlating HA and TGF-beta1 variations in cirrhosis patients (p = 0.018, r = 0.377). Fibrosis markers were unchanged at the end of the study in both groups. CONCLUSIONS: Olmesartan induced a mild reduction of portal pressure and TGF-beta1 for 1 year, but did not suppress hepatic fibrosis markers.

Natriuretic peptides and collagen biomarkers in patients with medical treatment for hypertension.

OBJECTIVE: The Anglo-Scandinavian Cardiac Outcomes Trial (ASCOT-BPLA) showed that an amlodipine-based regimen prevented more cardiovascular events than an atenolol-based regimen in patients at high risk of hypertension.The basis of this difference is partly unknown and may be due to their divergent effects on the remodelling process of hypertensive heart disease. METHODS AND RESULTS: We measured plasma levels of aminoterminal propeptide of atrial natriuretic peptide (NT-proANP) and aminoterminal propeptide of B-type natriuretic peptide and serum levels of the aminoterminal propeptide of type I procollagen (PINP), aminoterminal propeptide of type III procollagen and type I collagen telopeptide in 93 patients randomized in the ASCOT study at baseline and after two and four years and compared them with echocardiographic parameters and blood pressure. NT-proANP decreased at two years by 22 (-484 - 153) pmol/l in the amlodipine-based regimen and increased by 109 (-297 - 1545) pmol/l in the atenolol-based regimen (P < 0.001), whereas no significant difference in NT-proBNP between the arms was found. PINP levels increased by 1.8 (-29 -31) microg/l in the amlodipine-based regimen and decreased by 4.7 (-27- 31) microg/I in the atenolol-based regimen, whereas no differences were found in other collagen markers between the arms. Major echocardiographic changes were not found. CONCLUSIONS: Our results show that the two treatment regimens of ASCOT-BPLA had different effects on plasma natriuretic peptides and serological markers of collagen turnover, probably reflecting divergent effects in cardiac remodelling.

Bone turnover markers as predictive tools for skeletal complications in men with metastatic prostate cancer treated with zoledronic acid.

BACKGROUND: Bone turnover markers are helpful to diagnose bone metastases. The aim of this study was to evaluate the usefulness of these markers in prostate cancer patients with bone metastases before and during the treatment with zoledronic acid as predictive and monitoring tools of skeletal-related events (SRE). METHODS: One hundred seventeen prostate cancer patients with bone metastases and treated with zoledronic acid (4 mg every 4 weeks) were examined. Fifty-six patients were with and 61 patients without SRE during a 60-week study. Total and bone-specific alkaline phosphatase, and amino-terminal procollagen propeptides of type-I-collagen (PINP), cross-linked N-terminal (NTx), cross-linked C-terminal telopeptides of type-I-collagen (ICTP), and C-terminal telopeptides of type-I-collagen as well as prostate-specific antigen (PSA) were measured before and 12, 24, 36, 48, and 60 weeks after starting treatment. RESULTS: Higher baseline concentrations were observed in the SRE group. The bone markers except for ICTP and tALP decreased to 20-80% of the baseline values at week 12 after the drug administration showing a generally higher decline in the non-SRE group except for NTx. At all time points during treatment higher and increasing concentrations of bone markers were observed in the SRE group compared with non-SRE group. Cox regression models with clinical data and bone markers showed the baseline NTx concentration as predictor of SREs. During the study, percentage changes of PINP and ICTP were most indicative for SREs. CONCLUSIONS: Bone markers are useful tools to predict and diagnose SRE in prostate cancer patients with bone metastases under receiving zoledronic acid therapy.

Subantimicrobial-dose doxycycline modulates gingival crevicular fluid biomarkers of periodontitis in postmenopausal osteopenic women.

BACKGROUND: We recently demonstrated that a 2-year subantimicrobial-dose doxycycline (SDD) regimen (double-masked, placebo-controlled clinical trial) in postmenopausal (PM) women exhibiting mild systemic bone loss (osteopenia) and local bone loss (periodontitis) reduced the progression of periodontal attachment loss (intent-to-treat analysis) and the severity of gingival inflammation and alveolar bone loss (subgroups) without producing antibiotic side effects. We now describe SDD effects on biomarkers of collagen degradation and bone resorption in the gingival crevicular fluid (GCF) of the same vulnerable subjects. METHODS: GCF was collected from SDD- and placebo-treated PM subjects (n=64 each) at the baseline and 1- and 2-year appointments; the volume was determined; and the samples were analyzed for collagenase activity (using a synthetic peptide as substrate), relative levels of three genetically distinct collagenases (Western blot), a type-1 collagen breakdown product/bone resorption marker (a carboxyterminal telopeptide cross-link fragment of type I collagen [ICTP]; radioimmunoassay), and interleukin-1beta (enzyme-linked immunosorbent assay). Statistical analyses were performed using generalized estimating equations; primary analyses were intent-to-treat. RESULTS: Collagenase activity was significantly reduced by SDD treatment relative to placebo based on intent-to-treat (P=0.01). ICTP showed a similar pattern of change during SDD treatment, and GCF collagenase activity and ICTP were positively correlated at all time periods (P<0.001). Matrix metalloproteinase (MMP)-8 accounted for approximately 80% of total collagenase in GCF, with much less MMP-1 and -13, and SDD reduced the odds of elevated MMP-8 by 60% compared to placebo (P=0.006). CONCLUSION: These observations support the therapeutic potential of long-term SDD therapy to reduce periodontal collagen breakdown and alveolar bone resorption in PM women; effects on serum biomarkers of systemic bone loss in these subjects are being analyzed.

Amla (Emblica officinalis Gaertn.) extract promotes procollagen production and inhibits matrix metalloproteinase-1 in human skin fibroblasts.

AIM OF THE STUDY: Emblica officinalis Gaertn., commonly known as amla, is a rich dietary source of vitamin C, minerals and amino acids, and also contains various phenolic compounds. Amla extract is also known to exhibits potent antioxidant properties and to provide protection for human dermal fibroblasts against oxidative stress, and therefore it is thought to be useful for natural skin care. In this study, we investigated the effects of amla extract on human skin fibroblasts, especially for production of procollagen and matrix metalloproteinases (MMPs), in vitro. MATERIALS AND METHODS: Mitochondrial activity of human skin fibroblasts were measured by WST-8 assay. Quantification of procollagen, MMPs, and Tissue inhibitor of metalloproteinase-1 (TIMP-1) released from human skin fibroblasts were performed by immunoassay technique. RESULTS AND CONCLUSIONS: Amla extract stimulated proliferation of fibroblasts in a concentration-dependent manner, and also induced production of procollagen in a concentration- and time-dependent manner. Conversely, MMP-1 production from fibroblasts was dramatically decreased, but there was no evident effect on MMP-2. TIMP-1 was significantly increased by amla extract. From these results, it appears that amla extract works effectively in mitigative, therapeutic and cosmetic applications through control of collagen metabolism.

Effects of etanercept on serum biochemical markers of cartilage metabolism in patients with spondyloarthropathy.

OBJECTIVE: Anti-tumor necrosis factor (TNF) therapies provide symptomatic benefit in patients with spondyloarthropathy (SpA). Their effect on structural lesions has not yet been assessed. Biochemical markers of cartilage turnover revealing type II collagen degradation and synthesis are associated with joint damage in rheumatoid arthritis; their role in SpA is unknown. We describe the effects of etanercept on biochemical markers of type II collagen synthesis and degradation in patients with SpA followed for 2 years. METHODS: A total of 29 patients with SpA aged 22-68 years were included in a prospective 2-year study. Each patient received etanercept (25 mg twice a week) because of active disease despite optimal treatment. Cartilage degradation was investigated by measuring serum levels of the type II collagen fragments Helix-II and C2C, whereas the C-terminal propeptide of type II collagen (PIICP) was used as a marker of type II collagen synthesis. These markers were measured at baseline and after 1, 3, 6, 12, and 24 months of treatment. RESULTS: Over 2 years, there was a significant decrease of serum C2C (p = 0.0035 by repeated Friedman's test) and serum Helix-II (p = 0.004). Compared to baseline, the decrease of serum C2C was significant at Month 12 (-12.1%; p = 0.004), whereas the decrease of serum Helix-II was observed as early as 1 month (-18.1%; p = 0.015) after start of therapy, reaching a maximum decrease of -33.4% (p = 0.0079) at Month 12. Conversely, PIICP increased significantly by 17% (p = 0.006) at 24 months. CONCLUSION: These data suggest that etanercept may have beneficial effects on cartilage metabolism in patients with SpA.

Steroid treatment in ARDS: a critical appraisal of the ARDS network trial and the recent literature.

OBJECTIVES: To compare the design and results of randomized trials investigating prolonged glucocorticoid treatment (> or =7 days) in patients with acute lung injury-acute respiratory distress syndrome (ALI-ARDS), and review factors affecting response to therapy, including the role of secondary prevention. DESIGN: Trials were retrieved from the Cochrane Central Register of Controlled Trials (CENTRAL). Two investigators collected data on study characteristics, treatment intervention, and outcomes. The methodological quality of trials was determined and data were analyzed with Review Manager 4.2.3. MEASUREMENTS AND RESULTS: Five selected trials (n=518) consistently reported significant improvement in gas exchange, reduction in markers of inflammation, and decreased duration of mechanical ventilation and intensive care unit stay (all p<0.05). Two early small clinical trials showed marked reductions in the relative risk (RR) of death with glucocorticoid therapy (RR=0.14, 95% CI 0.04-0.53; p=0.004, I2=0%). Three subsequent larger trials, when combined, although nominally beneficial, did not reproduce the marked reductions observed in the earlier trials (RR=0.84; 95% CI 0.68-1.03; p=0.09, I2=9.1%), but achieved a distinct reduction in the RR of death in the larger subgroup of patients (n=400) treated before day 14 of ARDS [82/214 (38%) vs. 98/186 (52.5%), RR=0.78; 95% CI 0.64-0.96; p=0.02, I2=0%]. CONCLUSIONS: Prolonged glucocorticoid treatment substantially and significantly improves meaningful patient-centered outcome variables, and has a distinct survival benefit when initiated before day 14 of ARDS.

French maritime pine bark extract inhibits viral replication and prevents development of viral myocarditis.

BACKGROUND: French maritime pine bark extract (Pycnogenol) revealed diverse anti-inflammatory actions by an inhibition of NF-kappaB-dependent gene expression. The aim of this study was to determine whether Pycnogenol had a beneficial effect on viral myocarditis in mice. METHODS AND MATERIALS: Four-week-old inbred male DBA/2 mice were inoculated intraperitoneally with 10 plaque-forming units (pfu) of the encephalomyocarditis (EMC) virus. Pycnogenol was administered orally at a dose of 1 or 10 mg/kg per day for the histologic study, and 10 or 100 mg/kg for the gene expression study, beginning on the day of viral inoculation. RESULTS: The area of myocardial infiltration and necrosis on day 7 was significantly smaller in the hearts of mice treated with Pycnogenol 10 mg/kg (16.2 +/- 8.9% and 19.2 +/- 9.7%, respectively, n = 10, mean +/- SEM) compared with controls (27.6 +/- 15.0% and 30.1 +/- 15.7%, respectively, n = 10, P < .05). There was a nonsignificant trend for less myocardial infiltration in the Pycnogenol 1 mg/kg group. Myocardial virus concentration on day 7 was 8.4 +/- 0.3 x 10(3) pfu/mg in mice treated with 1 mg/kg of Pycnogenol, and 2.7 +/- 0.6 x 10(4) pfu/mg in control mice, and the difference was statistically significant (P < .05). Gene expressions of tumor necrosis factor, type-I procollagen, stem cell factor, and mast cell tryptase were significantly suppressed in the hearts of mice treated with Pycnogenol 100 mg/kg. CONCLUSIONS: These results suggest that Pycnogenol exerts its beneficial effects on viral myocarditis by decreasing virus replication, and by suppressing expression of pro-inflammatory cytokines, genes related to cardiac remodeling, and mast cell-related genes in the hearts of mice.

The effect of prior bisphosphonate exposure on the treatment response to teriparatide in clinical practice.

Our objective was to determine the effect of prior bisphosphonate exposure on the treatment response to teriparatide. All patients started on teriparatide in our hospital are entered into a database. All patients who had at least 12 months' treatment were identified. Patients were divided into two groups depending on whether or not they had prior bisphosphonate exposure, and the response to teriparatide was compared using procollagen of type 1 N-terminal propeptide (P1NP) and bone mineral density (BMD). Fifty-two patients had been treated for at least 12 months, 38 with prior bisphosphonate exposure and 14 without. The mean duration of bisphosphonate treatment was 67 months, discontinued a mean of 1 month previously. P1NP increased significantly at 3 and 6 months in both groups. However, those without previous bisphosphonate treatment had a higher baseline P1NP (49 vs. 30 microg/L, P<0.01), and this remained higher at 3 months (109 vs. 71 microg/L, P=0.10) and 6 months (183 vs. 126 microg/L, P=0.06), although the difference was not significant. In the prior bisphosphonate and bisphosphonate naive groups, respectively, the change in spinal BMD was 9.0% and 7.8% (P=0.54) at 12 months and 9.8% and 6.1% (P=0.30) at 18 months. The respective change in hip BMD was 1.0% and -0.3% (P=0.36) at 12 months and 2.8% and 1.3% (P=0.44) at 18 months. There was a trend toward a smaller but still significant increase in P1NP in response to teriparatide in bisphosphonate-treated patients. Although this suggests a blunting of the anabolic effects, in our clinic population this did not result in a reduction in BMD gain.