Faserverteilung und Muskelfaserspezifische glykolytische und oxidative Enzymaktivität im Skelettmuskel von Patienten mit Typ 2 Diabetes

Mittels immunhistochemischer- und zyto-fotometrischer Verfahren wurden die Änderungen der glykolytischen und der oxidativen Enzymkapazität im Skelettmuskel von Patienten mit T2DM mit der Muskelfasercharakteristik untersucht. Weiterführende Analysen klären den Zusammenhang zwischen der Änderung der Muskelfaserverteilung und der Enzymkapazität. Durch eine perkutane Biopsie des M. vastus lateralis wurden 10 Patienten mit T2DM und 15 Gesunden Probanden Muskelgewebe extrahiert. In der anschließenden Zytophotometrie erfolgte die Bestimmung der glykolytischen und oxidativen Enzymaktivität in Abhängigkeit der Fasercharakteristik nach SO, FOG und FT-Fasern. Die Untersuchung verdeutlichte eine Verminderung der oxidativen Enzymaktivität des M. vastus lateralis im Homogenat bei bestehenden T2DM mit gleichzeitiger Verringerung der SO- Muskelfasern um 16 Prozent und Erhöhung der FT- Muskelfasern um 49 Prozent im Vergleich zur Kontrollpopulation. Bei T2DM ist sowohl die oxidative als auch die glykolytische Enzymaktivität in den FG-Fasern als auch in den FOGMischfasern signifikant erhöht. Zusammenfassend weisen unsere Ergebnisse darauf hin, dass die geringere oxidative Enzymaktivität im Homogenat des Skelettmuskel von Patienten mit T2DM vielmehr durch einen verminderten Anteil von SO-Fasern als durch verminderte oxidative Aktivität in einzelnen Fasern verursacht ist. Die erhöhte glykolytische und oxidative Enzymaktivität in einzelnen Muskelfasern korreliert mit dem Maß langfristiger BZHomöostase und der Insulinempfindlichkeit. Diese Adaptation der Skelettmuskulatur könnte einen kompensatorischen Mechanismus bezüglich des pathologischen Glukosestoffwechsels des T2DM darstellen.:1. Zusammenfassung der Arbeiten 1.1 Hintergrund und Ziel der Arbeit 1.2 Studiendesign und Methoden 1.3 Ergebnisse 1.3.1 Bestimmung der Faserzusammensetzung und der metabolischen Enzymaktivität des M. vastus lateralis bei Typ 2 Diabetes mellitus 1.3.2 Zusammenhang zwischen den Parametern der Hyperglykämie und der Insulinresistenz mit der faserspezifischen Stoffwechselaktivität des M. vastus lateralis bei Typ 2 Diabetes mellitus 1.3.3 Die Stickstoffoxid-Synthase-Aktivität der Skelettmuskelfasern des Typ 2 Diabetes mellitus 1.4 Schlussfolgerungen 1.5 Literaturverzeichnis 2. Publikationen 2.1 Altered Fiber Distribution and Fiber-Specific Glycolytic and Oxidative Enzyme Activity in Skeletal Muscle of Patients With Type 2 Diabetes 2.2 Metabolic profile and nitric oxide synthase expression of skeletal muscle fibers are affected by type 1 and type 2 diabetes 3. Erklärung über die eigenständige Abfassung der Arbei

Nitric oxide synthase in skeletal muscle fibres of patients with type 2 diabetes

Muscle-derived nitric oxide (NO) mediates fundamental physiological actions on skeletal muscle including glucose uptake into muscle cells. Recently, we have shown that the altered glucose metabolism in skeletal muscle of patients with type 2 diabetes (T2D) is associated with changes in the metabolic profile of individual muscle fibres, but fibre-type specific changes in NO synthase (NOS) expression in skeletal muscle of T2D patients remain to be elucidated. Here we investigated fibre-type related NOS expression in vastus lateralis muscle of T2D patients compared with healthy individuals with normal glucose tolerance (NGT). Cytophotometrical assay and Western blotting did not reveal any quantitative differences between NOS expression in muscles from NGT and T2D subjects. Positive NOS immunoreactivity in vastus lateralis of T2D patients was found to be associated with fast-oxidative glycolytic (FOG) muscle phenotype. This indicates that NOS expression in T2D patients correlates both with skeletal muscle fibre type distribution and the activity of oxidative and glycolytic enzymes

Cytokine Effects on Gap Junction Communication and Connexin Expression in Human Bladder Smooth Muscle Cells and Suburothelial Myofibroblasts

BACKGROUND: The last decade identified cytokines as one group of major local cell signaling molecules related to bladder dysfunction like interstitial cystitis (IC) and overactive bladder syndrome (OAB). Gap junctional intercellular communication (GJIC) is essential for the coordination of normal bladder function and has been found to be altered in bladder dysfunction. Connexin (Cx) 43 and Cx45 are the most important gap junction proteins in bladder smooth muscle cells (hBSMC) and suburothelial myofibroblasts (hsMF). Modulation of connexin expression by cytokines has been demonstrated in various tissues. Therefore, we investigate the effect of interleukin (IL) 4, IL6, IL10, tumor necrosis factor-alpha (TNFα) and transforming growth factor-beta1 (TGFβ1) on GJIC, and Cx43 and Cx45 expression in cultured human bladder smooth muscle cells (hBSMC) and human suburothelial myofibroblasts (hsMF). METHODOLOGY/PRINCIPAL FINDINGS: HBSMC and hsMF cultures were set up from bladder tissue of patients undergoing cystectomy. In cytokine stimulated cultured hBSMC and hsMF GJIC was analyzed via Fluorescence Recovery after Photo-bleaching (FRAP). Cx43 and Cx45 expression was assessed by quantitative PCR and confocal immunofluorescence. Membrane protein fraction of Cx43 and Cx45 was quantified by Dot Blot. Upregulation of cell-cell-communication was found after IL6 stimulation in both cell types. In hBSMC IL4 and TGFβ1 decreased both, GJIC and Cx43 protein expression, while TNFα did not alter communication in FRAP-experiments but increased Cx43 expression. GJ plaques size correlated with coupling efficacy measured, while Cx45 expression did not correlate with modulation of GJIC. CONCLUSIONS/SIGNIFICANCE: Our finding of specific cytokine effects on GJIC support the notion that cytokines play a pivotal role for pathophysiology of OAB and IC. Interestingly, the effects were independent from the classical definition of pro- and antiinflammatory cytokines. We conclude, that connexin regulation involves genomic and/or post-translational events, and that GJIC in hBSMC and hsMF depend of Cx43 rather than on Cx45

Palmitate Induced IL-6 and MCP-1 Expression in Human Bladder Smooth Muscle Cells Provides a Link between Diabetes and Urinary Tract Infections

Therefore we studied the effects of the free fatty acid palmitate and bacterial lipopolysaccharide (LPS) on interleukin-6 (IL-6) and monocyte chemotactic protein-1 (MCP-1) expression and secretion in cultured human bladder smooth muscle cells (hBSMC).Biopsies were taken from patients undergoing cystectomy due to bladder cancer. Palmitate or LPS stimulated hBSMC were analysed for the production and secretion of the IL-6, gp80, gp80soluble, gp130, MCP-1, pSTAT3, SOCS3, NF-κB and SHP2 by quantitative PCR, ELISA, Western blotting, and confocal immunofluorescence. In signal transduction inhibition experiments we evaluated the involvement of NF-κB and MEK1 in IL-6 and MCP-1 regulation. Palmitate upregulates IL-6 mRNA expression and secretion via NF-κB dependent pathways in a concentration- and time-dependent manner. MCP-1 was moderately upregulated by palmitate but was strongly upregulated by LPS involving NF-κB and MEK1 dependent pathways. Soluble IL-6 receptor (gp80soluble) was downregulated by palmitate and LPS, while membrane-bound gp80 was moderately upregulated. LPS increased SOCS3 and SHP2, whereas palmitate only induced SOCS3. Secondary finding: most of the IL-6 is secreted.Bacterial infection (LPS) or metabolic alterations (palmitate) have distinct effects on IL-6 expression in hBSMC, (i) short term LPS induced autocrine JAK/STAT signaling and (ii) long-term endocrine regulation of IL-6 by palmitate. Induction of IL-6 in human bladder smooth muscle cells by fatty acids may represent a pathogenetic factor underlying the higher frequency and persistence of urinary tract infections in patients with metabolic diseases

Hybrid Surgery for Severe Mitral Valve Calcification: Limitations and Caveats for an Open Transcatheter Approach

Background and Objectives: Mitral stenosis with extensive mitral annular calcification (MAC) remains surgically challenging in respect to clinical outcome. Prolonged surgery time with imminent ventricular rupture and systolic anterior motion can be considered as a complex of causal factors. The aim of our alternative hybrid approach was to reduce the risk of annual rupture and paravalvular leaks and to avoid obstruction of the outflow tract. A review of the current literature was also carried out. Materials and Methods: Six female patients (mean age 76 +/- 9 years) with severe mitral valve stenosis and severely calcified annulus underwent an open implantation of an Edwards Sapien 3 prosthesis on cardiopulmonary bypass. Our hybrid approach involved resection of the anterior mitral leaflet, placement of anchor sutures and the deployment of a balloon expanded prosthesis under visual control. Concomitant procedures were carried out in three patients. Results: The mean duration of cross-clamping was 95 +/- 31 min and cardiopulmonary bypass was 137 +/- 60 min. The perioperative TEE showed in three patients an inconspicuous, heart valve-typical gradient on all implanted prostheses and a clinically irrelevant paravalvular leakage occurred in the anterior annulus. In the left ventricular outflow tract, mild to moderately elevated gradients were recorded. No adverse cerebrovascular events and pacemaker implantations were observed. All but one patient survived to discharge. Survival at one year was 83.3%. Conclusions: This off label implantation of the Edwards Sapien 3 prosthesis may be considered as a suitable bail-out approach for patients at high-risk for mitral valve surgery or deemed inoperable due to extensive MAC

Hybrid Surgery for Severe Mitral Valve Calcification: Limitations and Caveats for an Open Transcatheter Approach

Background and Objectives: Mitral stenosis with extensive mitral annular calcification (MAC) remains surgically challenging in respect to clinical outcome. Prolonged surgery time with imminent ventricular rupture and systolic anterior motion can be considered as a complex of causal factors. The aim of our alternative hybrid approach was to reduce the risk of annual rupture and paravalvular leaks and to avoid obstruction of the outflow tract. A review of the current literature was also carried out. Materials and Methods: Six female patients (mean age 76 9 years) with severe mitral valve stenosis and severely calcified annulus underwent an open implantation of an Edwards Sapien 3 prosthesis on cardiopulmonary bypass. Our hybrid approach involved resection of the anterior mitral leaflet, placement of anchor sutures and the deployment of a balloon expanded prosthesis under visual control. Concomitant procedures were carried out in three patients. Results: The mean duration of cross-clamping was 95 31 min and cardiopulmonary bypass was 137 60 min. The perioperative TEE showed in three patients an inconspicuous, heart valve-typical gradient on all implanted prostheses and a clinically irrelevant paravalvular leakage occurred in the anterior annulus. In the left ventricular outflow tract, mild to moderately elevated gradients were recorded. No adverse cerebrovascular events and pacemaker implantations were observed. All but one patient survived to discharge. Survival at one year was 83.3%. Conclusions: This “off label” implantation of the Edwards Sapien 3 prosthesis may be considered as a suitable bail-out approach for patients at high-risk for mitral valve surgery or deemed inoperable due to extensive MAC

Insufficient Oral Behaviour and the High Need for Periodontal Treatment in Patients with Heart Insufficiency and after Heart Transplantation: A Need for Special Care Programs?

Background: The aim of this cross-sectional study was the assessment of dental behaviour and oral health condition of heart transplant recipients (HTx) in comparison to patients with heart insufficiency (HI). Methods: Patients attending the Department for Cardiac Surgery, Leipzig Heart Center, Germany were recruited. Standardized questionnaires regarding dental behaviour and periodontal complaints were applied. A dental (decayed-, missing- and filled-teeth index) and periodontal examination (periodontal probing depth (PPD) and clinical attachment loss (CAL)) was performed. Based on the oral findings, dental and periodontal treatment need was determined. Statistics: T-test, Mann-Whitney U test, Chi-square test, and Fisher-test (p < 0.05). Results: A total of 201 patients (HTx: 112, HI: 89) were included. HTx patients were significantly more often allocated to dentists (p < 0.01). Furthermore, the HTx patients rated feeling informed appropriately about oral health more often (p < 0.01). HTx patients used interdental cleaning (p < 0.01) and mouth rinse (p = 0.02) more often than HI patients. No differences between groups were present regarding dental status and periodontitis severity (p > 0.05). Periodontal treatment need was high, showing prevalence of 79.5% (HTx) and 87.6% (HI, p = 0.14), respectively. Conclusions: Both groups show insufficient oral behaviour and a high need for periodontal treatment. Special care programs for HTx candidates and recipients appear recommendable

Lack in Periodontal Care of Patients Suffering from Severe Heart Diseases—Results after 12 Months Follow-Up

Background: To assess whether the standardized recommendation of patients with heart failure (HF), left-ventricular assist device (LVAD) and heart transplantation (HTx) to visit their dentist leads to improved oral conditions after 12 months. Methods: Patients from the Department of Cardiothoracic Surgery, Leipzig Heart Centre, Germany were examined at baseline and after 12 months. A dental (decayed-, missing-, and filled-teeth index (DMF-T)) and periodontal examination (periodontal probing depth, clinical attachment loss) was performed. At baseline, patients received a standardized recommendation to visit their dentist. At follow-up, a standardized questionnaire regarding the dental consultation was applied. Results: Eighty-eight participants (HTx: 31, LVAD: 43, HF: 14) were included. The majority of patients (79.5%) followed the recommendation to visit their dentist. Within the total cohort, periodontal treatment need was significantly reduced from 91% (baseline) to 75% (follow-up; p 0.05). Conclusions: The simple recommendation to visit the dentist appears not enough to obtain sufficient dental and periodontal conditions in patients with severe heart diseases. Thereby, a lack in periodontal treatment of patients with HF, HTx and LVAD was identified, making interdisciplinary dental special care programs recommendable

MRI assessment of changes in adipose tissue parameters after bariatric surgery

Bariatric surgery and other therapeutic options for obese patients are often evaluated by the loss of weight, reduction of comorbidities or improved quality of life. However, little is currently known about potential therapy-related changes in the adipose tissue of obese patients. The aim of this study was therefore to quantify fat fraction (FF) and T1 relaxation time by magnetic resonance imaging (MRI) after Roux-en-Y gastric bypass surgery and compare the resulting values with the preoperative ones. Corresponding MRI data were available from 23 patients (16 females and 7 males) that had undergone MRI before (M0) and one month after (M1) bariatric surgery. Patients were 22-59 years old (mean age 44.3 years) and their BMI ranged from 35.7-54.6 kg/m(2) (mean BMI 44.6 kg/m(2)) at M0. Total visceral AT volumes (VVAT-T, in L) were measured by semi-automatic segmentation of axial MRI images acquired between diaphragm and femoral heads. MRI FF and T1 relaxation times were measured in well-defined regions of visceral (VAT) and subcutaneous (SAT) adipose tissue using two custom-made analysis tools. Average BMI values were 45.4 kg/m(2) at time point M0 and 42.4 kg/m(2) at M1. Corresponding VVAT-T values were 5.94 L and 5.33 L. Intraindividual differences in both BMI and VVAT-T were highly significant (p<0.001). Average relaxation times T1 VAT were 303.7 ms at M0 and 316.9 ms at M1 (p<0.001). Corresponding T1(SAT) times were 283.2 ms and 280.7 ms (p = 0.137). Similarly, FFVAT differences (M0: 85.7%, M1: 83.4%) were significant (p <0.01) whereas FFSAT differences (M0: 86.1, M1: 85.9%) were not significant (p = 0.517). In conclusion, bariatric surgery is apparently not only related to a significant reduction in common parameters of adipose tissue distribution, here BMI and total visceral fat volume, but also significant changes in T1 relaxation time and fat fraction of visceral adipose tissue. Such quantitative MRI measures may potentially serve as independent biomarkers for longitudinal and cross-sectional measurements in obese patients

Exercise Training Prevents Diaphragm Contractile Dysfunction in Heart Failure

Purpose: Patient studies have demonstrated the efficacy of exercise training in attenuating respiratory muscle weakness in chronic heart failure (HF), yet direct assessment of muscle fiber contractile function together with data on the underlying intracellular mechanisms remains elusive. The present study, therefore, used a mouse model of HF to assess whether exercise training could prevent diaphragm contractile fiber dysfunction by potentially mediating the complex interplay between intracellular oxidative stress and proteolysis. Methods: Mice underwent sham operation (n = 10) or a ligation of the left coronary artery and were randomized to sedentary HF (n = 10) or HF with aerobic exercise training (HF + AET; n = 10). Ten weeks later, echocardiography and histological analyses confirmed HF. Results: In vitro diaphragm fiber bundles demonstrated contractile dysfunction in sedentary HF compared with sham mice that was prevented by AET, with maximal force 21.0 ± 0.7 versus 26.7 ± 1.4 and 25.4 ± 1.4 N·cm−2, respectively (P < 0.05). Xanthine oxidase enzyme activity and MuRF1 protein expression, markers of oxidative stress and protein degradation, were ~20% and ~70% higher in sedentary HF compared with sham mice (P < 0.05) but were not different when compared with the HF + AET group. Oxidative modifications to numerous contractile proteins (i.e., actin and creatine kinase) and markers of proteolysis (i.e., proteasome and calpain activity) were elevated in sedentary HF compared with HF + AET mice (P < 0.05); however, these indices were not significantly different between sedentary HF and sham mice. Antioxidative enzyme activities were also not different between groups. Conclusion: Our findings demonstrate that AET can protect against diaphragm contractile fiber dysfunction induced by HF, but it remains unclear whether alterations in oxidative stress and/or protein degradation are primarily responsible