Distribution patterns of intramyocellular and extramyocellular fat by magnetic resonance imaging in subjects with diabetes, prediabetes and normoglycaemic controls.

AIM: To evaluate the distribution of intramyocellular lipids (IMCLs) and extramyocellular lipids (EMCLs) as well as total fat content in abdominal skeletal muscle by magnetic resonance imaging (MRI) using a dedicated segmentation algorithm in subjects with type 2 diabetes (T2D), prediabetes and normoglycaemic controls. MATERIALS AND METHODS: Subjects from a population-based cohort were classified with T2D, prediabetes or as normoglycaemic controls. Total myosteatosis, IMCLs and EMCLs were quantified by multiecho Dixon MRI as proton-density fat-fraction (in %) in abdominal skeletal muscle. RESULTS: Among 337 included subjects (median age 56.0 [IQR: 49.0-64.0] years, 56.4% males, median body mass index [BMI]: 27.2 kg/m2 ), 129 (38.3%) were classified with an impaired glucose metabolism (T2D: 49 [14.5%]; prediabetes: 80 [23.7%]). IMCLs were significantly higher than EMCLs in subjects without obesity (5.7% [IQR: 4.8%-7.0%] vs. 4.1% [IQR: 2.7%-5.8%], P < .001), whereas the amounts of IMCLs and EMCLs were shown to be equal and significantly higher in subjects with obesity (both 6.7%, P < .001). Subjects with prediabetes and T2D had significantly higher amounts of IMCLs and EMCLs compared with normoglycaemic controls (P < .001). In univariable analysis, prediabetes and T2D were significantly associated with both IMCLs (prediabetes: ß: 0.76, 95% CI: 0.28-1.24, P = .002; T2D: ß: 1.56, 95% CI: 0.66-2.47, P < .001) and EMCLs (prediabetes: ß: 1.54, 95% CI: 0.56-2.51, P = .002; T2D: ß: 2.15, 95% CI: 1.33-2.96, P < .001). After adjustment for age and gender, the association of IMCLs with prediabetes attenuated (P = 0.06), whereas for T2D, both IMCLs and EMCLs remained significantly and positively associated (P < .02). CONCLUSION: There are significant differences in the amount and distribution ratio of IMCLs and EMCLs between subjects with T2D, prediabetes and normoglycaemic controls. Therefore, these patterns of intramuscular fat distribution by MRI might serve as imaging biomarkers in both normal and impaired glucose metabolism.

Mitochondrial dysfunction in human immunodeficiency virus-1 transgenic mouse cardiac myocytes.

The pathophysiology of human immunodeficiency virus (HIV)-associated cardiomyopathy remains uncertain. We used HIV-1 transgenic (Tg26) mice to explore mechanisms by which HIV-related proteins impacted on myocyte function. Compared to adult ventricular myocytes isolated from nontransgenic (wild type [WT]) littermates, Tg26 myocytes had similar mitochondrial membrane potential (ΔΨ m ) under normoxic conditions but lower Δ Ψ m after hypoxia/reoxygenation (H/R). In addition, Δ Ψ m in Tg26 myocytes failed to recover after Ca 2+ challenge. Functionally, mitochondrial Ca 2+ uptake was severely impaired in Tg26 myocytes. Basal and maximal oxygen consumption rates (OCR) were lower in normoxic Tg26 myocytes, and further reduced after H/R. Complex I subunit and ATP levels were lower in Tg26 hearts. Post-H/R, mitochondrial superoxide (O 2•- ) levels were higher in Tg26 compared to WT myocytes. Overexpression of B-cell lymphoma 2-associated athanogene 3 (BAG3) reduced O 2•- levels in hypoxic WT and Tg26 myocytes back to normal. Under normoxic conditions, single myocyte contraction dynamics were similar between WT and Tg26 myocytes. Post-H/R and in the presence of isoproterenol, myocyte contraction amplitudes were lower in Tg26 myocytes. BAG3 overexpression restored Tg26 myocyte contraction amplitudes to those measured in WT myocytes post-H/R. Coimmunoprecipitation experiments demonstrated physical association of BAG3 and the HIV protein Tat. We conclude: (a) Under basal conditions, mitochondrial Ca 2+ uptake, OCR, and ATP levels were lower in Tg26 myocytes; (b) post-H/R, Δ Ψ m was lower, mitochondrial O 2•- levels were higher, and contraction amplitudes were reduced in Tg26 myocytes; and (c) BAG3 overexpression decreased O 2•- levels and restored contraction amplitudes to normal in Tg26 myocytes post-H/R in the presence of isoproterenol.

Associations of myosteatosis with disc degeneration: A 3T magnetic resonance imaging study in individuals with impaired glycaemia.

BACKGROUND: Intervertebral disc degeneration (IVDD) may be linked to dysregulations of skeletal muscle glucose metabolism and fatty alterations of muscle composition (Myosteatosis). Our aim was to evaluate the different associations of magnetic resonance imaging (MRI)-based paravertebral myosteatosis with lumbar disc degeneration in individuals with impaired glucose metabolism and normoglycaemic controls. METHODS: In total, 304 individuals (mean age: 56.3 ± 9.1 years, 53.6% male sex, mean body mass index [BMI]: 27.6 ± 4.7 kg/m2 ) from a population-based cohort study who underwent 3-Tesla whole-body chemical-shift-encoded (six echo times) and T2-weighted single-shot-fast-spin-echo MRI were included. Lumbar disc degeneration was assessed at motion segments L1 to L5, categorized according to the Pfirrmann score and defined as Pfirrmann grade > 2 and/or disc bulging/herniation on at least one segment. Fat content of the autochthonous back muscles and the quadratus lumborum muscle was quantified as proton density fat fraction (PDFFmuscle ). Logistic regression models adjusted for age, sex, BMI and regular physical activity were calculated to evaluate the association between PDFFmuscle and outcome IVDD. RESULTS: The overall prevalence of IVDD was 79.6%. There was no significant difference in the prevalence or severity distribution of IVDD between participants with or without impaired glucose metabolism (77.7% vs. 80.7%, P = 0.63 and P = 0.71, respectively). PDFFmuscle was significantly and positively associated with an increased risk for the presence of IVDD in participants with impaired glycaemia when adjusted for age, sex and BMI (PDFFautochthonous back muscles : odds ratio [OR] 2.16, 95% confidence interval [CI] [1.09, 4.3], P = 0.03; PDFFquadratus lumborum : OR 2.01, 95% CI [1.04, 3.85], P = 0.04). After further adjustment for regular physical activity, the results attenuated, albeit approaching statistical significance (PDFFautochthonous back muscles : OR 1.97, 95% CI [0.97, 3.99], P = 0.06; PDFFquadratus lumborum : OR 1.86, 95% CI [0.92, 3.76], P = 0.09). No significant associations were shown in healthy controls (PDFFautochthonous back muscles : OR 0.62, 95% CI [0.34, 1.14], P = 0.13; PDFFquadratus lumborum : OR 1.06, 95% CI [0.6, 1.89], P = 0.83). CONCLUSIONS: Paravertebral myosteatosis is positively associated with intervertebral disc disease in individuals with impaired glucose metabolism, independent of age, sex and BMI. Regular physical activity may confound these associations. Longitudinal studies will help to better understand the pathophysiological role of skeletal muscle in those with concomitant disturbed glucose haemostasis and intervertebral disc disease, as well as possible underlying causal relationships.

Population-based cohort imaging: skeletal muscle mass by magnetic resonance imaging in correlation to bioelectrical-impedance analysis.

BACKGROUND: Skeletal muscle mass is subjected to constant changes and is considered a good predictor for outcome in various diseases. Bioelectrical-impedance analysis (BIA) and magnetic resonance imaging (MRI) are approved methodologies for its assessment. However, muscle mass estimations by BIA may be influenced by excess intramuscular lipids and adipose tissue in obesity. The objective of this study was to evaluate the feasibility of quantitative assessment of skeletal muscle mass by MRI as compared with BIA. METHODS: Subjects from a population-based cohort underwent BIA (50 kHz, 0.8 mA) and whole-body MRI including chemical-shift encoded MRI (six echo times). Abdominal muscle mass by MRI was quantified as total and fat-free cross-sectional area by a standardized manual segmentation-algorithm and normalized to subjects' body height2 (abdominal muscle mass indices: AMMIMRI ). RESULTS: Among 335 included subjects (56.3 ± 9.1 years, 56.1% male), 95 (28.4%) were obese (BMI ≥ 30 kg/m2 ). MRI-based and BIA-based measures of muscle mass were strongly correlated, particularly in non-obese subjects [r < 0.74 in non-obese (P < 0.001) vs. r < 0.56 in obese (P < 0.001)]. Median AMMITotal(MRI) was significantly higher in obese as compared with non-obese subjects (3246.7 ± 606.1 mm2 /m2 vs. 2839.0 ± 535.8 mm2 /m2 , P < 0.001, respectively), whereas the ratio AMMIFat-free /AMMITotal (by MRI) was significantly higher in non-obese individuals (59.3 ± 10.1% vs. 53.5 ± 10.6%, P < 0.001, respectively). No significant difference was found regarding AMMIFat-free(MRI) (P = 0.424). In analyses adjusted for age and sex, impaired glucose tolerance and measures of obesity were significantly and positively associated with AMMITotal(MRI) and significantly and inversely with the ratio AMMIFat-free(MRI) /AMMITotal(MRI) (P < 0.001). CONCLUSIONS: MRI-based assessment of muscle mass is feasible in population-based imaging and strongly correlated with BIA. However, the observed weaker correlation in obese subjects may explain the known limitation of BIA in obesity and promote MRI-based assessments. Thus, skeletal muscle mass parameters by MRI may serve as practical imaging biomarkers independent of subjects' body weight.

Healthcare and forensic medical aspects of police detainees, suspects and complainants in Europe.

Death and harm is well-recognised in detainees in police custody worldwide. Based on the results of previous global surveys and the CPT (European Committee for the Prevention of Torture) recommendations a questionnaire was developed to summarise the current medical aspects of police custody in European countries. The survey was distributed to named contacts in all European countries. Data from 25 European countries was obtained. The results reveal significant differences in the regulations among the different countries, with nothing close to a harmonised European standard in place at present. This study has identified interesting variations in the methods and standards of healthcare and forensic medical services to detainees in police custody (e.g. quantitative mode of monitoring, qualification of the doctors, maximum time allowed for holding a detainee in police custody, body or an organisation that investigates complaints against the police). There are both very detailed legal regulations in some countries while in others there are only generally observed provisions that sometimes are only given in the form of recommendations. A multinational, multiprofessional expert group is required to identify best practices, recommend basic standards of care and identify qualifications which would be appropriate for healthcare professionals working in this field.

Inter- and intra-observer variability of an anatomical landmark-based, manual segmentation method by MRI for the assessment of skeletal muscle fat content and area in subjects from the general population.

OBJECTIVES: Changes in skeletal muscle composition, such as fat content and mass, may exert unique metabolic and musculoskeletal risks; however, the reproducibility of their assessment is unknown. We determined the variability of the assessment of skeletal muscle fat content and area by MRI in a population-based sample. METHODS: A random sample from a prospective, community-based cohort study (KORA-FF4) was included. Skeletal muscle fat content was quantified as proton-density fat fraction (PDFF) and area as cross-sectional area (CSA) in multi-echo Dixon sequences (TR 8.90 ms, six echo times, flip angle 4°) by a standardized, anatomical landmark-based, manual skeletal muscle segmentation at level L3 vertebra by two independent observers. Reproducibility was assessed by intraclass correlation coefficients (ICC), scatter and Bland-Altman plots. RESULTS: From 50 subjects included (mean age 56.1 ± 8.8 years, 60.0% males, mean body mass index 28.3 ± 5.2) 2'400 measurements were obtained. Interobserver agreement was excellent for all muscle compartments (PDFF: ICC0.99, CSA: ICC0.98) with only minor absolute and relative differences (-0.2 ± 0.5%, 31 ± 44.7 mm2; -2.6 ± 6.4% and 2.7 ± 3.9%, respectively). Intra-observer reproducibility was similarly excellent (PDFF: ICC1.0, 0.0 ± 0.4%, 0.4%; CSA: ICC1.0, 5.5 ± 25.3 mm2, 0.5%, absolute and relative differences, respectively). All agreement was independent of age, gender, body mass index, body height and visceral adipose tissue (ICC0.96-1.0). Furthermore, PDFF reproducibility was independent of CSA (ICC0.93-0.99).  Conclusions:  Quantification of skeletal muscle fat content and area by MRI using an anatomical landmark-based, manual skeletal muscle segmentation is highly reproducible. Advances in knowledge: An anatomical landmark-based, manual skeletal muscle segmentation provides high reproducibility of skeletal muscle fat content and area and may therefore serve as a robust proxy for myosteatosis and sarcopenia in large cohort studies.

Assessment of the degree of abdominal myosteatosis by magnetic resonance imaging in subjects with diabetes, prediabetes and healthy controls from the general population.

OBJECTIVES: Intra- and intermyocellular lipid deposition and adipose tissue are part of glucose homeostasis and insulin resistance; however, their role in type 2 diabetes mellitus (T2DM) remains unclear. We assessed differences in the degree of abdominal myosteatosis among subjects with T2DM and prediabetes. MATERIALS AND METHODS: Asymptomatic subjects from the general population were classified as subjects with T2DM, prediabetes or healthy controls and underwent multi-echo Dixon magnetic resonance imaging (MRI) (TR 8.90 ms, six echo times, flip-angle 4°). Abdominal myosteatosis was quantified as proton-density fat-fraction (PDFFmuscle) by a standardized segmentation-algorithm. Cardiometabolic risk factors were prospectively obtained in a comprehensive health assessment and visceral and subcutaneous adipose tissue (VAT and SAT) were quantified semi-automatically. Uni- and multivariate quantile regression were used to examine associations. RESULTS: Among 349 included subjects (mean age: 56.0 ±â€¯8.0years, 56.7% males), 45 were classified as subjects with T2DM and 84 with prediabetes (12.9% and 24.1%; respectively). Median PDFFmuscle was significantly higher in subjects with T2DM and prediabetes compared to healthy controls (13.1% (IQR10.5-16.6%); 11.1% (IQR8.9-15.0%) and 10.1% (IQR7.5-13.3%); respectively, p < 0.001). The observed differences were independent of age and gender (all p < 0.002) but attenuated after adjustment for BMI (ß: -0.02, 95%CI: -1.49 to 1.44, p = 0.974; ß: 0.47, 95%CI: -0.91 to 1.86, p = 0.506; prediabetes and T2DM, respectively). This effect was attributable to VAT, which remained independently associated with PDFFmuscle after full adjustment (ß: 0.01, 95%CI: 0.01-0.02, p = 0.002). CONCLUSIONS: There are significant differences in the degree of abdominal myosteatosis between subjects with T2DM, prediabetes and healthy controls, that may be confounded by VAT. However, abdominal myosteatosis by MRI might serve as a cardiometabolic imaging-biomarker, specifically in the setting of impaired glucose metabolism.

Methylene Blue Counteracts H2S-Induced Cardiac Ion Channel Dysfunction and ATP Reduction.

We have previously demonstrated that methylene blue (MB) counteracts the effects of hydrogen sulfide (H2S) cardiotoxicity by improving cardiomyocyte contractility and intracellular Ca2+ homeostasis disrupted by H2S poisoning. In vivo, MB restores cardiac contractility severely depressed by sulfide and protects against arrhythmias, ranging from bundle branch block to ventricular tachycardia or fibrillation. To dissect the cellular mechanisms by which MB reduces arrhythmogenesis and improves bioenergetics in myocytes intoxicated with H2S, we evaluated the effects of H2S on resting membrane potential (Em), action potential (AP), Na+/Ca2+ exchange current (INaCa), depolarization-activated K+ currents and ATP levels in adult mouse cardiac myocytes and determined whether MB could counteract the toxic effects of H2S on myocyte electrophysiology and ATP. Exposure to toxic concentrations of H2S (100 µM) significantly depolarized Em, reduced AP amplitude, prolonged AP duration at 90% repolarization (APD90), suppressed INaCa and depolarization-activated K+ currents, and reduced ATP levels in adult mouse cardiac myocytes. Treating cardiomyocytes with MB (20 µg/ml) 3 min after H2S exposure restored Em, APD90, INaCa, depolarization-activated K+ currents, and ATP levels toward normal. MB improved mitochondrial membrane potential (∆ψm) and oxygen consumption rate in myocytes in which Complex I was blocked by rotenone. We conclude that MB ameliorated H2S-induced cardiomyocyte toxicity at multiple levels: (1) reversing excitation-contraction coupling defects (Ca2+ homeostasis and L-type Ca2+ channels); (2) reducing risks of arrhythmias (Em, APD, INaCa and depolarization-activated K+ currents); and (3) improving cellular bioenergetics (ATP, ∆ψm).