Prevalence, incidence and concomitant co-morbidities of type 2 diabetes mellitus in South Western Germany--a retrospective cohort and case control study in claims data of a large statutory health insurance.

BACKGROUND: Type 2 diabetes mellitus (T2DM) has become a world-wide epidemic. This chronic metabolic disease has a major impact on life expectancy and on quality of life. The burden of this disease includes a number of co-morbidities. However, estimates of prevalence, incidence and associated diseases as well as the current temporal development and regional differences are largely missing for South Western Germany. METHODS: Lifetime diagnosis-based prevalence, incidence and presence of concomitant co-morbidities were examined between the years 2007 and 2010 in the claims data set of all insured persons of the AOK Baden-Wuerttemberg, a large statutory health insurance. The analysis was based on the respective WHO-ICD-10 codes. Data were standardized for age and sex on the residential population of about 10 million inhabitants of South Western Germany. RESULTS: The total study cohort involved approximately 3.5 million persons each year. The standardized diagnosis-based prevalence (SDP) of T2DM rose from 6.6%, 7.4%, 8.0%, up to 8.6% in the years 2007 to 2010. Yearly SDP was between 14.0% and 18.9% at an age range of 60 to 64 years and between 26.7% and 31.8% at an age of 75 years or older. In the year 2010 the regional distributions of standardized diagnosis-based prevalence were between 7.6% and 11.6 %, respectively. Incidence rates were 8.3 in 2008, 7.8 in 2009, and 8.7 in 2010 (all rates per 1000). The excess disease risk (odds ratio) of T2DM was for adiposity 2.8 to 3.0, hypertension 2.4 to 3.7, coronary heart disease 1.8 to 1.9, stroke 1.7 to 1.8, renal insufficiency 2.8 to 3.4, and retinopathy 2.8 to 2.9 in the years 2007 to 2010. These co-morbidities appeared several years earlier compared to the non-diabetic population. CONCLUSIONS: T2DM is common and increasing in South Western Germany. In particular a quarter of the population in higher ages was afflicted by T2DM. Interestingly a region-specific pattern was observed as well as an increase in numbers during earlier years in life. Our data underline the need for diabetes awareness programmes including early diagnosis measures as well as structured and timely health surveys for major diseases such as T2DM and its concomitant co-morbidities.

Proteome scale turnover analysis in live animals using stable isotope metabolic labeling.

At present most quantitative proteomics investigations are focused on the analysis of protein expression differences between two or more sample specimens. With each analysis a static snapshot of a cellular state is captured with regard to protein expression. However, any information on protein turnover cannot be obtained using classic methodologies. Protein turnover, the result of protein synthesis and degradation, represents a dynamic process, which is of equal importance to understanding physiological processes. Methods employing isotopic tracers have been developed to measure protein turnover. However, applying these methods to live animals is often complicated by the fact that an assessment of precursor pool relative isotope abundance is required. Also, data analysis becomes difficult in case of low label incorporation, which results in a complex convolution of labeled and unlabeled peptide mass spectrometry signals. Here we present a protein turnover analysis method that circumvents this problem using a (15)N-labeled diet as an isotopic tracer. Mice were fed with the labeled diet for limited time periods and the resulting partially labeled proteins digested and subjected to tandem mass spectrometry. For the interpretation of the mass spectrometry data, we have developed the ProTurnyzer software that allows the determination of protein fractional synthesis rates without the need of precursor relative isotope abundance information. We present results validating ProTurnyzer with Escherichia coli protein data and apply the method to mouse brain and plasma proteomes for automated turnover studies.

Pentraxin 3 is elevated in haemodialysis patients and is associated with cardiovascular disease.

BACKGROUND: Pentraxins are mediators of inflammation as well as markers of the acute-phase reaction. While elevation of C-reactive protein (CRP) in patients with renal failure and its association with cardiovascular disease is well described, there are no data on pentraxin 3 (PTX3) in this population. METHODS: Plasma was obtained from 44 chronic haemodialysis (HD) patients, 35 peritoneal dialysis (PD) patients, 39 patients with chronic renal failure (CRF) not on dialysis therapy and 14 age-matched normal subjects. PTX3 production in whole blood was also investigated in samples taken before and during HD. RESULTS: PTX3 plasma levels were significantly higher in HD patients (5.8 +/- 0.6 ng/ml) compared with the other three groups. There were no significant differences between PD patients (1.5 +/- 0.4 ng/ml), CRF patients (1.5 +/- 0.4 ng/ml) and normal subjects (0.76 +/- 0.2 ng/ml). In dialysis patients, PTX3 levels correlated significantly with time on renal replacement therapy (RRT) and with weekly erythropoietin dose. PTX3 levels were significantly higher in patients with coronary artery disease and peripheral artery disease compared with those without. During a single HD session, PTX3 production was higher in whole blood samples taken after 3 h HD compared with samples taken before HD. CONCLUSIONS: PTX3 levels are markedly elevated in HD patients. The increase in PTX3 production in whole blood after HD indicates that the HD procedure itself contributes to elevated PTX3 levels in HD patients. The association between PTX3 and cardiovascular morbidity suggests a possible connection of PTX3 with atherosclerosis and cardiovascular disease in HD patients.

Prevalence of silent gastric ulcer, erosions or severe acute gastritis in patients with type 2 diabetes mellitus--a cross-sectional study.

BACKGROUND/AIMS: Severe gastric inflammation or ulcer disease can alter gastric motility and influence sufficient glycemic control in patients with type 2 diabetes mellitus. However, visceral neuropathy may reduce the perception of typical gastrointestinal symptoms in these patients. The aim of the present study was to evaluate the prevalence of silent severe acute gastritis, gastric ulcers or erosions in asymptomatic patients with diabetes mellitus and to determine potential predictive parameters. METHODOLOGY: Seventy-two patients with type 2 diabetes mellitus and little or no dyspeptic symptoms were investigated by endoscopy of the upper gastrointestinal tract under screening conditions. Before endoscopy the presence of gastrointestinal symptoms and standard laboratory parameters were determined. In addition, the presence of Helicobacter pylori infection was investigated by rapid urease test and histology. RESULTS: Highly active gastric inflammation was found in 34 patients (gastric ulcers in 10, gastric erosions in 21, and histologically acute, grade two or three gastritis in 3 patients). Episodic heartburn was significantly associated with highly active gastric inflammation (odds ratio 2.96 (1.05-8.32), p = 0.036). Elevated levels of C-reactive-protein and blood leukocyte counts proved to be of positive predictive value for highly active gastric inflammation in patients without other causes of acute inflammatory diseases (odds ratio 3.52 (p = 0.026) and 7.64 (p = 0.007) respectively). No significant association was found for gender, age, duration of disease, BMI, considerably raised HbA1c (>8.5%), complications of diabetic disease, general gastrointestinal symptoms, Helicobacter pylori infections and therapy with acetylsalicylic acid on 100 mg/d. CONCLUSIONS: The results of this study indicate that severe acute gastric inflammation or ulcer disease can occur with high prevalence in patients with diabetes mellitus with little or no dyspeptic symptoms. Additional endoscopic investigations might be of particular diagnostic value in patients with inexplicable raised levels of inflammatory parameters like C-reactive-protein or blood leukocyte counts.

Decrease in circulating endothelial cell adhesion molecule and thrombomodulin levels during oral iloprost treatment in rheumatoid arthritis patients: preliminary results.

OBJECTIVE: Rheumatoid arthritis is a chronic inflammatory autoimmune disease with proinflammatory cytokines involved in its pathogenesis. Recently in vitro as well as in vivo studies have shown that iloprost, a stable prostacyclin analogue, can reduce the release of these cytokines. This study was performed to further investigate the anti-inflammatory effects of iloprost by determining plasma adhesion molecules as markers of endothelial cell activation, and plasma thrombomodulin as a parameter of endothelial cell injury in patients with rheumatoid arthritis receiving oral iloprost therapy. METHODS: Plasma thrombomodulin levels and the values of the plasma adhesion molecules VCAM-1 (vascular cell adhesion molecule 1), E-selectin (CD62E), and ICAM-1 (intercellular adhesion molecule 1, CD 54) were measured by ELISA during a 7-day period of treatment with orally-administered iloprost in 14 patients with active rheumatoid arthritis. Finally, the same parameters were determined at the end of the observation period (1 week after the end of therapy). In addition, the disease activity was measured using the DAS (disease activity score) as well as the patients' self-assessed pain severity, and correlated with the changes of plasma adhesion molecule and thrombomodulin levels. RESULTS: The plasma levels of all three adhesion molecules as well as of thrombomodulin significantly decreased under therapy with oral iloprost. After 1 week (day 7 of therapy), the mean percent changes from day 0 were -20.1% for VCAM-1 (p = 0.008), -21.2 for ICAM-1 (p = 0.003), -24.6% for E-selectin (p = 0.001), and -21.7% for thrombomodulin (p = 0.003). This decrease lasted up to 1 week after the end of therapy in the case of VCAM-1 (p = 0.023) and ICAM-1 (p = 0.001). Further analysis of the results revealed additional significant correlations between different parameters of clinical disease activity, thrombomodulin and adhesion molecules. CONCLUSION: This study showed hints towards clinical effects in patients with rheumatoid arthritis receiving oral iloprost therapy. Pathophysiologically, the decrease of adhesion molecules points at an immunomodulating effect of iloprost. The observed thrombomodulin-lowering effect of iloprost may indicate stabilisation of endothelial cell function by diminishing endothelial cell injury.

Comparison of three commercially available thrombomodulin ELISA kits.

Thrombomodulin is a transmembranous glycoprotein of endothelial cells. In vitro it is a marker of endothelial cell injury. In vivo the levels of soluble serum thrombomodulin are regarded as parameters of disease activity in vasculitides and vasculopathies. However, the mean thrombomodulin values of different studies show marked concentration differences of the control values. The purpose of this study was to further investigate these differences. We examined 60 sera of patients with systemic lupus erythematosus (SLE) and 10 of healthy controls with three commercially available thrombomodulin ELISA kits for determination of their thrombomodulin concentration and correlation to disease activity. The disease activity of the SLE patients was determined with the SLAM-score. Raised thrombomodulin values were found in 58% (test A), 55% (test B) and 61.6% (test C). The thrombomodulin values significantly correlated with the SLE disease activity independently of the ELISA kit used (correlation coefficients: r=0.84 (test A), r=0.80 (test B), and r=0.65 (test C)). In addition, the correlation coefficients between the respective thrombomodulin values of the three tests were r=0.86 (test A to B), r=0.73 (test A to C) and r=0.79 (test B to C). However, significant differences between the results of the three ELISA kits were found between the detected thrombomodulin concentrations. The mean thrombomodulin concentrations of the controls were 25.6 ng/ml (test A), 3.53 ng/ml (test B), and 2.52 ng/ml (test C). Our results reveal that the soluble thrombomodulin values of all three commercially available ELISA kits significantly correlate with the disease activity of SLE patients. However, the results show significant differences in the determined thrombomodulin concentrations. A calibration would be required of the different ELISA kits in order to permit a direct comparison of the results of these thrombomodulin ELISAs. A general reference standard would be desirable for this calibration of all thrombomodulin ELISAs. However, this general reference standard has to be adapted to the distinct test conditions of all test kits as well as including all epitopes of thrombomodulin which are recognised by the different antibodies used in the respective test kits. At present, only ELISA kits from the same manufacturer should be used during a single study including any follow-up investigations.

Comparison of a 48-hour infusion of 5-fluorouracil without folinic acid with 24-hour folinic acid/5-fluorouracil in patients with metastatic colorectal cancer refractory to bolus folinic acid/5-fluorouracil. A prospective cohort study.

BACKGROUND: Out of various high-dose 5-fluorouracil (5-FU) regimens given with or without folinic acid (FA), the optimal 5-FU schedule has still to be defined as treatment for metastatic colorectal cancer (CRC). Consequently, we compared toxicity, response and survival following two FA/5-FU regimens in 55 CRC patients refractory to bolus FA/5-FU. METHODS: Twenty-eight patients (group A) received 5-FU (60 mg/kg body weight) for 48 h, and 27 (group B) received 2-hour infusions of FA (500 mg/m(2)) and 24-hour infusions of 5-FU (2600 mg/m(2)) until disease progression. RESULTS: Both groups were adequately matched with respect to patient characteristics. While overall toxicities were rare, hand-foot syndrome was more common in A. Tumor control was achieved in 57 and 44%, for A and B, respectively. Survival times were 16 months in A and 9 months in B. CONCLUSIONS: Since both 5-FU infusion protocols showed equivalent palliative effects, FA may be questioned in second-line 5-FU regimens.

Kinetics of thrombomodulin release and endothelial cell injury by neutrophil-derived proteases and oxygen radicals.

Thrombomodulin is a transmembranous glycoprotein of endothelial cells. In vitro it is a marker of endothelial cell injury. In vivo the levels of serum thrombomodulin are regarded as a parameter of activity in vasculitides. The latter are pathophysiologically determined by neutrophil-derived inflammation and endothelial cell injury caused by secretion of proteases and hydrogen peroxide. It was the objective of this study to determine whether thrombomodulin is only a late marker of advanced endothelial cell injury or whether it indicates also earlier stages of cell alterations. Over 24 hr endothelial cell cultures were incubated with hydrogen peroxide or the neutrophil proteases proteinase-3, elastase and cathepsin G. The time-dependent increase of thrombomodulin in the supernatant was determined by enzyme-linked immunosorbent assay and immunoblot. In addition the viability (eosin, tetrazolium dye assay), detachment (crystal-violet assay), and apoptosis (4',6-diamine-2'-phenylindole-dihydrochloride assay) of the respective endothelial cells were determined for adherent and non-adherent cells. A rapid thrombomodulin increase was found under all experimental conditions. The additional immunoblotting analysis showed the pattern of proteolytic cleavage caused by the protease reactivity. In case of hydrogen peroxide the thrombomodulin increase was closely correlated with the loss of cell viability and lysis. The incubation of endothelial cells with the different proteases resulted in a time-dependent detachment of primarily viable cells. In addition to cell necrosis apoptotic cell death was found in the subgroup of detached endothelial cells after prolonged incubation over 24 hr with proteinase-3 (23%), elastase (31%), and cathepsin G (19%). In contrast, still adhering cells did not show any signs of necrosis or apoptosis. In summary these studies confirm in vitro that soluble thrombomodulin is not only a parameter of advanced endothelial cell destruction itself but also in addition an early marker of initial endothelial cell membrane changes induced by neutrophil derived proteases and oxygen radicals.

A 7-day oral treatment of patients with active rheumatoid arthritis using the prostacyclin analog iloprost: cytokine modulation, safety, and clinical effects.

OBJECTIVE: The purpose of this study was to investigate the plasma levels of tumor necrosis factor (TNF)-alpha, its soluble p55 and p75 receptors, ex vivo lipopolysaccharide (LPS)-stimulated TNF-alpha production, and plasma levels of interleukin 6, interleukin 1, and interleukin 10 during a 7-day oral administration of iloprost in patients with active rheumatoid arthritis. METHODS: During oral 7-day administration of the prostacyclin analog iloprost, the plasma levels of TNF-alpha, soluble p55, and p75 TNF-alpha receptors, IL-1, IL-6, and IL-10 and C-reactive protein (CRP) in serum were determined on days -1, 1, 3, 4, 6, and 8 and after a treatment-free follow-up on day 15. In addition, the ex vivo TNF-alpha production in whole blood under LPS-stimulated and -unstimulated conditions were measured. Fifteen patients with active rheumatoid arthritis and baseline TNF-alpha plasma levels of > or =2 pg/ml were included in independent groups receiving 50 microg, 100 microg, or 150 microg iloprost per day in addition to their conventional antirheumatic therapy. The respective dose was given once daily from days 1 to 3 and doubled from days 4 to 7. The tender and swollen joint count (28 joints) and the patients' assessments of pain severity and general feeling (10-cm visual analog scale) were performed on days -1, 4, and 8 (end of treatment) and after a 7-day follow-up. RESULTS: The patients showed decreased TNF-alpha levels during iloprost administration. The decrease in the ex vivo LPS-stimulated TNF-alpha production and plasma levels of the p75 TNF receptor were found to be associated with a decrease in the number of tender joints. Additionally, IL-6 was downregulated. CONCLUSION: A 7-day oral administration of iloprost resulted in a change of in vivo and ex vivo cellular cytokine production, with reductions in TNF-alpha and p75 TNF receptor plasma levels. These changes were associated with clinical improvements in active rheumatoid arthritis.