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In persons with dyslipidemia, a high residual risk of cardiovascular disease remains despite lipid lowering therapy. Current cardiovascular risk prediction mainly focuses on low-density lipoprotein cholesterol (LDL-c) levels, neglecting other contributing risk factors. Moreover, the efficacy of LDL-c lowering by statins resulting in reduced cardiovascular risk is only partially effective. Secondly, from a metrological viewpoint LDL-c falls short as a reliable measurand. Both direct and calculated LDL-c tests produce inaccurate test results at the low end under aggressive lipid lowering therapy. As LDL-c tests underperform both clinically and metrologically, there is an urging need for molecularly defined biomarkers. Over the years, apolipoproteins have emerged as promising biomarkers in the context of cardiovascular disease as they are the functional workhorses in lipid metabolism. Among these, apolipoprotein B (ApoB), present on all atherogenic lipoprotein particles, has demonstrated to clinically outperform LDL-c. Other apolipoproteins, such as Apo(a) - the characteristic apolipoprotein of the emerging risk factor lipoprotein(a) -, and ApoC-III - an inhibitor of triglyceride-rich lipoprotein clearance -, have attracted attention as well. To support personalized medicine, we need to move to molecularly defined risk markers, like the apolipoproteins. Molecularly defined diagnosis and molecularly targeted therapy require molecularly measured biomarkers. This review provides a summary of the scientific validity and (patho)physiological role of nine serum apolipoproteins, Apo(a), ApoB, ApoC-I, ApoC-II, ApoC-III, ApoE and its phenotypes, ApoA-I, ApoA-II, and ApoA-IV, in lipid metabolism, their association with cardiovascular disease, and their potential as cardiovascular risk markers when measured in a multiplex apolipoprotein panel. [ABSTRACT FROM AUTHOR]

Right‐sided myocardial mechanical efficiency (work output/metabolic energy input) in pulmonary hypertension can be severely reduced. We determined the contribution of intrinsic myocardial determinants of efficiency using papillary muscle preparations from monocrotaline‐induced pulmonary hypertensive (MCT‐PH) rats. The hypothesis tested was that efficiency is reduced by mitochondrial dysfunction in addition to increased activation heat reported previously. Right ventricular muscle preparations were subjected to 5 Hz sinusoidal length changes at 37°C. Work and suprabasal oxygen consumption (V̇O2${\dot{V}}_{{{\rm{O}}}_{\rm{2}}}$) were measured before and after cross‐bridge inhibition by blebbistatin. Cytosolic cytochrome c concentration, myocyte cross‐sectional area, proton permeability of the inner mitochondrial membrane and monoamine oxidase and glucose 6‐phosphate dehydrogenase activities and phosphatidylglycerol/cardiolipin contents were determined. Mechanical efficiency ranged from 23% to 11% in control (n = 6) and from 22% to 1% in MCT‐PH (n = 15) and correlated with work (r2 = 0.68, P < 0.0001) but not with V̇O2${\dot{V}}_{{{\rm{O}}}_{\rm{2}}}$ (r2 = 0.004, P = 0.7919). V̇O2${\dot{V}}_{{{\rm{O}}}_{\rm{2}}}$ for cross‐bridge cycling was proportional to work (r2 = 0.56, P = 0.0005). Blebbistatin‐resistant V̇O2${\dot{V}}_{{{\rm{O}}}_{\rm{2}}}$ (r2 = 0.32, P = 0.0167) and proton permeability of the mitochondrial inner membrane (r2 = 0.36, P = 0.0110) correlated inversely with efficiency. Together, these variables explained the variance of efficiency (coefficient of multiple determination r2 = 0.79, P = 0.0001). Cytosolic cytochrome c correlated inversely with work (r2 = 0.28, P = 0.0391), but not with efficiency (r2 = 0.20, P = 0.0867). Glucose 6‐phosphate dehydrogenase, monoamine oxidase and phosphatidylglycerol/cardiolipin increased in the right ventricular wall of MCT‐PH but did not correlate with efficiency. Reduced myocardial efficiency in MCT‐PH is a result of activation processes and mitochondrial dysfunction. The variance of work and the ratio of activation heat reported previously and blebbistatin‐resistant V̇O2${\dot{V}}_{{{\rm{O}}}_{\rm{2}}}$ are discussed. Key points: Mechanical efficiency of right ventricular myocardium is reduced in pulmonary hypertension. Increased energy use for activation processes has been demonstrated previously, but the contribution of mitochondrial dysfunction is unknown.Work and oxygen consumption are determined during work loops. Oxygen consumption for activation and cross‐bridge cycling confirm the previous heat measurements.Cytosolic cytochrome c concentration, proton permeability of the mitochondrial inner membrane and phosphatidylglycerol/cardiolipin are increased in experimental pulmonary hypertension.Reduced work and mechanical efficiency are related to mitochondrial dysfunction.Upregulation of the pentose phosphate pathway and a potential gap in the energy balance suggest mitochondrial dysfunction in right ventricular overload is a resiult of the excessive production of reactive oxygen species. [ABSTRACT FROM AUTHOR]

An elevated low-density lipoprotein cholesterol concentration is a classical risk factor for cardiovascular disease. This has led to pharmacotherapy in patients with atherosclerotic heart disease or high heart disease risk with statins to reduce serum low-density lipoprotein cholesterol. Even in patients in whom the target levels of low-density lipoprotein cholesterol are reached, there remains a significant residual cardiovascular risk; this is due, in part, to a focus on low-density lipoprotein cholesterol alone and neglect of other important aspects of lipoprotein metabolism. A more refined lipoprotein analysis will provide additional information on the accumulation of very low-density lipoproteins, intermediate density lipoproteins, chylomicrons, chylomicron-remnants and Lp(a) concentrations. Instead of measuring the cholesterol and triglyceride content of the lipoproteins, measurement of their apolipoproteins (apos) is more informative. Apos are either specific for a particular lipoprotein or for a group of lipoproteins. In particular measurement of apos in atherogenic particles is more biologically meaningful than the measurement of the cholesterol concentration contained in these particles. Applying apo profiling will not only improve characterization of the lipoprotein abnormality, but will also improve definition of therapeutic targets. Apo profiling aligns with the concept of precision medicine by which an individual patient is not treated as 'average' patient by the average (dose of) therapy. This concept of precision medicine fits the unmet clinical need for stratified cardiovascular medicine. The requirements for clinical application of proteomics, including apo profiling, can now be met using robust mass spectrometry technology which offers desirable analytical performance and standardization. [ABSTRACT FROM AUTHOR]

A wide variety of protein post‐translational modifications (PTMs) decorate cellular proteins, regulating their structure, interactions and ultimately their function. The density of co‐occurring PTMs on proteins can be very high, where multiple PTMs can positively or negatively influence each other's actions, termed PTM crosstalk. In this review, we highlight recent progress in the area of PTM crosstalk, whereby we focus on crosstalk between protein phosphorylation and O‐GlcNAcylation. These two PTMs largely target identical (i.e., Ser and Thr) amino acids in proteins. Phosphorylation/O‐GlcNAcylation crosstalk comes in many flavors, for instance by competition for the same site/residue (reciprocal crosstalk), as well as by modifications influencing each other in proximity or even distal on the protein sequence. PTM crosstalk is observed on the writers of these modifications (i.e., kinases and O‐GlcNAc transferase), on the erasers (i.e., phosphatases and O‐GlcNAcase), and on the readers and the substrates. We describe examples of all these different flavors of crosstalk, and additionally the methods that are emerging to better investigate in particular phosphorylation/O‐GlcNAcylation crosstalk. [ABSTRACT FROM AUTHOR]

Optimizing physical performance is a major goal in current physiology. However, basic understanding of combining high sprint and endurance performance is currently lacking. This study identifies critical determinants of combined sprint and endurance performance using multiple regression analyses of physiologic determinants at different biologic levels. Cyclists, including 6 international sprint, 8 team pursuit, and 14 road cyclists, completed a Wingate test and 15-km time trial to obtain sprint and endurance performance results, respectively. Performance was normalized to lean body mass2/3 to eliminate the influence of body size. Performance determinants were obtained from whole-body oxygen consumption, blood sampling, knee-extensor maximal force, muscle oxygenation, whole-muscle morphology, and muscle fiber histochemistry of musculus vastus lateralis. Normalized sprint performance was explained by percentage of fast-type fibers and muscle volume (R² = 0.65; P < 0.001) and normalized endurance performance by performance oxygen consumption (Vo2), mean corpuscular hemoglobin concentration, and muscle oxygenation (R² = 0.92; P < 0.001). Combined sprint and endurance performance was explained by gross efficiency, performance Vo2, and likely by muscle volume and fascicle length (P = 0.056; P = 0.059). High performance Vo2 related to a high oxidative capacity, high capillarization × myoglobin, and small physiologic cross-sectional area (R² = 0.67; P < 0.001). Results suggest that fascicle length and capillarization are important targets for training to optimize sprint and endurance performance simultaneously. [ABSTRACT FROM AUTHOR]

Purpose Graphical optimization (GrO) is a common method for high-dose-rate/pulsed-dose-rate (PDR) prostate brachytherapy treatment planning. New methods performing inverse optimization of the dose distribution have been developed over the past years. The purpose is to compare GrO and two established inverse methods, inverse planning simulated annealing (IPSA) and hybrid inverse treatment planning and optimization (HIPO), and one new method, enhanced geometric optimization–interactive inverse planning (EGO–IIP), in terms of speed and dose–volume histogram (DVH) parameters. Methods and Materials For 26 prostate cancer patients treated with a PDR brachytherapy boost, an experienced treatment planner optimized the dose distributions using four different methods: GrO, IPSA, HIPO, and EGO–IIP. Relevant DVH parameters (prostate— V 100% , D 90% , V 150% ; urethra— D 0.1cm3 and D 1.0cm3 ; rectum— D 0.1cm3 and D 2.0cm3 ; bladder— D 2.0cm3 ) were evaluated and their compliance to the constraints. Treatment planning time was also recorded. Results All inverse methods resulted in shorter planning time (mean, 4–6.7 min), as compared with GrO (mean, 7.6 min). In terms of DVH parameters, none of the inverse methods outperformed the others. However, all inverse methods improved on compliance to the planning constraints as compared with GrO. On average, EGO–IIP and GrO resulted in highest D 90% , and the IPSA plans resulted in lowest bladder D 2.0cm3 and urethra D 1.0cm3 . Conclusions Inverse planning methods decrease planning time as compared with GrO for PDR/high-dose-rate prostate brachytherapy. DVH parameters are comparable for all methods. [ABSTRACT FROM AUTHOR]

Key points Skeletal muscles increase oxygen consumption to produce energy during exercise; however, the processes controlling the rate of adaptation (its kinetics) at exercise onset and offset are not well understood., Here we measure kinetics in single frog skeletal muscle fibres using a unique experimental system that allows features of intracellular control mechanisms to be elucidated., We show for the first time that at contractions onset skeletal muscle kinetics are best described by a biphasic 'activation' and 'exponential' profile, whereas at cessation recovers with a single smooth exponential., Additionally these features were dependent on oxidative capacity and the intensity of stimulated contractions., These data show that the intracellular processes that activate oxidative energy provision at the onset of contractions are far more complex than previously suggested., Abstract The mechanisms controlling skeletal muscle oxygen consumption () during exercise are not well understood. We determined whether first-order control could explain kinetics at contractions onset () and cessation () in single skeletal muscle fibres differing in oxidative capacity, and across stimulation intensities up to . Xenopus laevis fibres ( n= 21) were suspended in a sealed chamber with a fast response electrode to measure every second before, during and after stimulated isometric contractions. A first-order model did not well characterise on-transient kinetics. Including a time delay (TD) in the model provided a significantly improved characterisation than a first-order fit without TD ( F-ratio; P < 0.05), and revealed separate 'activation' and 'exponential' phases in 15/21 fibres contracting at (mean ± SD TD: 14 ± 3 s). On-transient kinetics () was weakly and linearly related to ( R2= 0.271, P= 0.015). Off-transient kinetics, however, were first-order, and was greater in low-oxidative ( < 0.05 nmol mm−3 s−1) than high-oxidative fibres ( > 0.10 nmol mm−3 s−1; 170 ± 70 vs. 29 ± 6 s, P < 0.001). was proportional to ( R2= 0.727, P < 0.001), unlike in the on-transient. The calculated oxygen deficit was larger ( P < 0.05) than the post-contraction volume of consumed oxygen at all intensities except . These data show a clear dissociation between the kinetic control of at the onset and cessation of contractions and across stimulation intensities. More complex models are therefore required to understand the activation of mitochondrial respiration in skeletal muscle at the start of exercise. [ABSTRACT FROM AUTHOR]

Background: Patients with complicated intra-abdominal infections are prone to develop multiple organ failure, including myocardial dysfunction. We hypothesized that early dysfunction during sepsis is associated with inflammation, mitochondrial injury, impaired mitochondrial function, and activation of mitochondrial biogenesis. Materials and Methods: Rats received lipopolysaccharide (LPS, n = 11) intraperitoneally. Healthy rats (n = 6) served as controls. Myocardial function was measured ex vivo in an isolated Langendorff-perfused heart set-up. Myocardial vascular cell adhesion molecule-1 (VCAM-1) expression was determined by immunofluorescence microscopy. Cytochrome c release and cytochrome c oxidase (COX IV) activity were measured by immunohistochemistry and enzyme histochemistry, respectively. Protein expression of tumor necrosis factor-α (TNF-α), B-cell lymphoma (Bcl)-2, peroxisome proliferator activated receptor γ cofactor 1α (PGC-1α), and mitochondrial transcription factor A (TFAM) were analyzed by Western blot technique. Mitochondria were studied by electron microscopy. Results: Two hours after LPS injection, developed pressure had decreased and after 4 h myocardial contractility (+dP/dt) and relaxation (–dP/dt) also had decreased. TNF-α protein expression was increased after 2 h and returned to normal at 4 h, whereas after 4 h VCAM-1 expression was higher in LPS-treated animals. At 2 h a substrate-dependent increase in COXIV-activity was seen, but no mitochondrial damage occurred as cytochrome c release, COX IV activity and Bcl-2, PGC-1α or TFAM expression were not changed. Electron microscopy did not reveal differences in myocardial mitochondrial characteristics between LPS-treated and control rats. Conclusions: Early myocardial dysfunction in sepsis is associated with myocardial inflammation but not with mitochondrial injury, impaired mitochondrial function, or activated mitochondrial biogenesis. [Copyright &y& Elsevier]

Ir-192 wires have been used in low-dose-rate brachytherapy for many years. Commercially available treatment planning systems approximate the dose rate distribution of the straight or curved wires applying the superposition principle using one of the following methods: (i) The wire is modeled as a set of point sources, (ii) the wire is modeled as a set of small straight segment wires, (iii) the values of the parameters and functions of the American Association of Physicists in Medicine (AAPM) Task Group 43 protocol are obtained for wire lengths between 3 and 7 cm assuming some simplifications. The dose rate distributions obtained using these methods for linear wires of different lengths and U-shaped wires present significant deviations compared to those obtained by Monte Carlo. In the present study we propose a new method to model 192Ir wires of any length and shape, named the Two Lengths based Segmented method. This method uses the formalism stated in the AAPM Task Group 43 protocol for two straight wires only, 0.5 and 1 cm, to obtain the dose rate distribution around wires of any length (down to 0.3 cm and up to 10 cm) improving on the results of the aforementioned ones. This method can easily be applied to dose calculations around other wires, such as Pd-103 ones. [ABSTRACT FROM AUTHOR]

A hypothesis is presented that explains one of the mechanisms by which a heart starts to fail. The hypothesis is that myocardial function of an overloaded or otherwise stressed heart may become impaired by cellular troponin degradation in vital cardiomyocytes. The troponins (I, T and C) regulate actin–myosin interaction, thereby controlling contraction and relaxation. Troponins have been shown to be targets of activated calpain I. This enzyme, that is activated by elevated intracellular Ca2+ concentrations, such as occurs during ischemia, degrades troponins, leading to impaired interaction between actin and myosin and, thereby, less contractile force. Several reports about troponin degradation in viable myocardium support this hypothesis. Also, results are discussed that demonstrate the presence of immunoreactive troponin fragments in plasma under conditions in which myocardial necrosis can be excluded or is unlikely. The hypothesis implicates that release of troponin and/or troponin degradation products is not specific for necrotic myocardium but may occur from viable myocardium as well. To test this hypothesis, several lines of research are suggested. If the hypothesis is not rejected in the near future, the concept that a positive troponin test reflects ‘even microscopic zones of myocardial necrosis’ as used by the Joint ESC/ACC Committee for the Redefinition of Myocardial Infarction [The Joint European Society of Cardiology/American College of Cardiology Committee for the Redefinition of Myocardial Infarction. Myocardial infarction redefined—A consensus document of the Joint European Society of Cardiology/American College of Cardiology Committee for the Redefinition of Myocardial Infarction. Eur Heart J 2000;21:1502–1513], should be withdrawn. [Copyright &y& Elsevier]

Focuses on a study which measured the oxygen consumption and heat production during contraction and recovery of isolated white muscle fibers from dogfish, Scyliorhinus canicula. Materials and methods; Results of the study; Discussion.

Provides information on a study which evaluated the short-term effect of dietary counselling in patients with endogenous hypertriglyceridemia and evaluated the effects of advised nutrient changes. Design of the study; Methods used in the study; How dietary counselling in patients with endogenous hypertriglyceridemia effectively improve lipid and lipoprotein levels.

Studies the origin of the threefold variation in isometric force normalized to cross-sectional area of single fast-twitch tibialis anterior muscle fibers of the frog Rana temporaria. Stimulus protocol and variation between frogs; Effects of artificial hibernation on normalized force.

Introduction: To investigate the additive prognostic value of growth differentiation factor (GDF-15) levels in ST-segment elevation myocardial infarction (STEMI) patients treated with primary percutaneously coronary intervention (pPCI) with 10-year mortality on top of clinical characteristics and known cardiac biomarkers. Methods: Baseline serum GDF-15 levels were measured in 290 STEMI patients treated with pPCI in the MISSION! intervention trial conducted from February 1, 2004 through October 31, 2006. The incremental prognostic value of GDF-15 and NTproBNP levels was evaluated on top of clinical characteristics using Cox proportional hazards analysis, Chi-square models and C-index. Outcome was 10-year all-cause mortality. Results: Mean age was 59.0 ± 11.5 years and 65 (22.4) patients were female. A total of 37 patients died during a follow-up of 9.4 (IQR 8.8–10.0) years. Multivariable Cox regression revealed GDF-15 and NTproBNP levels above median to be independently associated with 10-year all-cause mortality [HR GDF-15, 2.453 (95% CI 1.064–5.658), P = 0.04; HR NTproBNP, 2.413 (95% CI 1.043–5.564), P = 0.04] after correction for other clinical variables. Stratified by median GDF-15 (37.78 pmol/L) and NTproBNP (11.74 pmol/L) levels, Kaplan–Meier curves showed significant better survival for patients with GDF-15 and NTproBNP levels below the median versus above the median. The likelihood ratio test showed a significant incremental value of GDF-15 (P = 0.03) as compared with a model with clinically important variables and NTproBNP. The C-statistics for this model improved from 0.82 to 0.84 when adding GDF-15. Conclusion: GDF-15 levels at admission in STEMI patients are independently associated with 10-year all-cause mortality rates and could improve risk stratification on top of clinical variables and other cardiac biomarkers. [ABSTRACT FROM AUTHOR]

Myocardial efficiency of pulmonary hypertensive (PH) patients and isolated right ventricular papillary muscles from PH rats can be severely reduced. We aimed to determine the causes of the efficiency reduction in isolated papillary muscles of monocrotaline-injected PH rats (MCT, 60mg/kg; 8 control and 16 MCT male Wistar rats). After 3 weeks PH was confirmed by echocardiography and right ventricular papillary muscles were isolated. Work and suprabasal oxygen consumption were determined using the work loop technique: 4 min 5 Hz sinusoidal length changes at 92.5% of optimum length (L0), amplitude 0.15 L0 in 1.5 mM Ca2+ Tyrode solution at 37°C. Morphological and enzymehistochemical characteristics of the muscle and free wall were also quantified. Cardiolipin content was determined by HPLC-MS. Passive tension (overall m±SD: 23.9±9.3 mN/mm2) and active tension at 0.5Hz (75±28 mN/mm2 myocyte) were similar in control and MCT muscles at L0. Net efficiency of control muscles was 15.5±4.9 % and correlated with cross-sectional area of myocytes (273±88 μm2) in the muscle (r=0.84, P=0.017), indicating that larger myocytes are more efficient. However, net efficiency of MCT muscles was lower (9.4±7.4%, P=0.027) and did not correlate with myocyte cross-sectional area (448±88μm2). The stimulus phase for maximum work was 40±12° in control and differed (P=0.019) from 27±9° in MCT muscles, indicating slower force development in MCT muscles. Power output per loop tended to be lower in MCT muscles (1.24±0.56 versus 0.83±0.75 μJ/mm3 myocyte, P=0.16). Work loops after crossbridge inhibition by blebbistatin (10μM) were -0.38±0.22 μJ/mm3, indicating similar viscoelastic properties of control and MCT muscles. The stimulus phase for maximum work correlated with work (r=0.72, P=0.002) and efficiency (r=0.65, P=0.006) in MCT muscles, but not with myofibrillar CaATPase activity. Oxygen consumption during stimulation after blebbistatin correlated negatively with efficiency (r=-0.60, P=0.006), indicating increased oxygen consumption for non-contractile reactions in MCT muscles. Oligomycin-resistant MgATPase activity increased (P=0.004) whereas cardiolipin 72:8 decreased (P=0.023) in right-sided myocardium of MCT, indicating changes of the mitochondrial inner membrane. Monoamine oxidase A activity (P=0.004) and glucose-6-phosphate dehydrogenase activity (P=0.0004) in myocytes increased, indicating oxidative stress and increased oxygen consumption other than oxidative phosphorylation. These results demonstrate that metabolic rather than sarcomeric changes cause reduced myocardial efficiency in experimental pulmonary hypertension. [ABSTRACT FROM PUBLISHER]

The function of the right ventricle (RV) determines the prognosis of patients with pulmonary hypertension. While much progress has been made in the treatment of pulmonary hypertension, therapies for the RV are less well established. In this review of treatment strategies for the RV, first we focus on ways to reduce wall stress since this is the main determinant of changes to the ventricle. Secondly, we discuss treatment strategies targeting the detrimental consequences of increased RV wall stress. To reduce wall stress, afterload reduction is the essential. Additionally, preload to the ventricle can be reduced by diuretics, by atrial septostomy, and potentially by mechanical ventricular support. Secondary to ventricular wall stress, left-to-right asynchrony, altered myocardial energy metabolism, and neurohumoral activation will occur. These may be targeted by optimising RV contraction with pacing, by iron supplement, by angiogenesis and improving mitochondrial function, and by neurohumoral modulation, respectively. We conclude that several treatment strategies for the right heart are available; however, evidence is still limited and further research is needed before clinical application can be recommended. [ABSTRACT FROM AUTHOR]

Myoglobin is essential for oxygen transport to the muscle fibers. However, measurements of myoglobin (Mb) protein concentrations within individual human muscle fibers are scarce. Recent observations have revealed surprisingly low Mb concentrations in elite cyclists, however it remains unclear whether this relates to Mb translation, transcription and/or myonuclear content. The aim was to compare Mb concentration, Mb messenger RNA (mRNA) expression levels and myonuclear content within muscle fibers of these elite cyclists with those of physically-active controls. Muscle biopsies were obtained from m. vastus lateralis in 29 cyclists and 20 physically-active subjects. Mb concentration was determined by peroxidase staining for both type I and type II fibers, Mb mRNA expression level was determined by quantitative PCR and myonuclear domain size (MDS) was obtained by immunofluorescence staining. Average Mb concentrations (mean ± SD: 0.38 ± 0.04 mM vs. 0.48 ± 0.19 mM; P = 0.014) and Mb mRNA expression levels (0.067 ± 0.019 vs. 0.088 ± 0.027; P = 0.002) were lower in cyclists compared to controls. In contrast, MDS and total RNA per mg muscle were not different between groups. Interestingly, in cyclists compared to controls, Mb concentration was only lower for type I fibers (P < 0.001), but not for type II fibers (P > 0.05). In conclusion, the lower Mb concentration in muscle fibers of elite cyclists is partly explained by lower Mb mRNA expression levels per myonucleus and not by a lower myonuclear content. It remains to be determined whether cyclists may benefit from strategies that upregulate Mb mRNA expression levels, particularly in type I fibers, to enhance their oxygen supply. [ABSTRACT FROM AUTHOR]

A letter to the editor is presented in response to the article "Reduced Systemic Oxygen Extraction Does Not Prove Muscle Dysfunction in PAH," by James J. Tolle and colleagues.

Examines the basic energetic characteristics of heart muscle activation and contraction in mouse. Proportion of mouse genes without a human homologue; Oxygen consumption of the mouse heart; Way to prevent hypoxia.

Purpose: The aims of this study were (i) to design a new high-dose-rate (HDR) brachytherapy applicator for treating surface lesions with planning target volumes larger than 3 cm in diameter and up to 5 cm in size, using the microSelectron-HDR or Flexitron afterloader (Elekta Brachytherapy) with a 192Ir source; (ii) to calculate by means of the Monte Carlo(MC) method the dose distribution for the new applicator when it is placed against a water phantom; and (iii) to validate experimentally the dose distributions in water. Methods: The PENELOPE2008MC code was used to optimize dwell positions and dwell times. Next, the dose distribution in a water phantom and the leakage dose distribution around the applicator were calculated. Finally, MC data were validated experimentally for a 192Ir mHDR-v2 source by measuring (i) dose distributions with radiochromic EBT3 films (ISP); (ii) percentage depth-dose (PDD) curve with the parallel-plate ionization chamber Advanced Markus (PTW); and (iii) absolute dose rate with EBT3 films and the PinPoint T31016 (PTW) ionization chamber. Results: The new applicator is made of tungsten alloy (Densimet) and consists of a set of interchangeable collimators. Three catheters are used to allocate the source at prefixed dwell positions with preset weights to produce a homogenous dose distribution at the typical prescription depth of 3 mm in water. The same plan is used for all available collimators. PDD, absolute dose rate per unit of air kerma strength, and off-axis profiles in a cylindrical water phantom are reported. These data can be used for treatment planning. Leakage around the applicator was also scored. The dose distributions, PDD, and absolute dose rate calculated agree within experimental uncertainties with the doses measured: differences of MC data with chamber measurements are up to 0.8% and with radiochromic films are up to 3.5%. Conclusions: The new applicator and the dosimetric data provided here will be a valuable tool in clinical practice, making treatment of large skin lesions simpler, faster, and safer. Also the dose to surrounding healthy tissues is minimal. [ABSTRACT FROM AUTHOR]

Abstract: Background: Despite international standardization programs for LDLc and HDLc measurements, results vary significantly with methods from different manufacturers. We aimed to simulate the impact of analytical error and hypertriglyceridemia on HDLc- and LDLc-based cardiovascular risk classification. Methods: From the Dutch National EQA-2012 external quality assessment of 200 clinical laboratories, we examined data from normotriglyceridemic (∼1 mmol/l) and hypertriglyceridemic (∼7 mmol/l) serum pools with lipid target values assigned by the Lipid Reference Laboratory in Rotterdam. HDLc and LDLc were measured using direct methods of Abbott, Beckman, Siemens, Roche, Olympus, or Ortho Clinical Diagnostics. We simulated risk reclassification using HDL- and sex-specific SCORE multipliers considering two fictitious moderate-risk patients with initial SCORE 4% (man) and 3% (woman). Classification into high-risk treatment groups (LDLc >2.50 mmol/l) was compared between calculated LDLc and direct LDLc methods. Results: Overall HDLc measurements in hypertriglyceridemic serum showed negative mean bias of −15%. HDL-multipliers falsely reclassified 70% of women and 43% of men to a high-risk (SCORE >5%) in hypertriglyceridemic serum (P < 0.0001 vs. normotriglyceridemic serum) with method-dependent risk reclassifications. Direct LDLc in hypertriglyceridemic serum showed positive mean bias with Abbott (+16%) and Beckman (+14%) and negative mean bias with Roche (−7%). In hypertriglyceridemic serum, 57% of direct LDLc measurements were above high-risk treatment goal (2.50 mmol/l) vs. 29% of direct LDLc (33% of calculated LDLc) in normotriglyceridemic sera. Conclusion: LDLc and HDLc measurements are unreliable in severe hypertriglyceridemia, and should be applied with caution in SCORE risk classification and therapeutic strategies. [Copyright &y& Elsevier]

We investigated whether the lower fatigue resistance in smokers than in nonsmokers is caused by a compromised muscle oxidative metabolism. Using calibrated histochemistry, we found no differences in succinate dehydrogenase (SDH) activity, myoglobin concentration, or capillarization in sections of the vastus lateralis muscle between smokers and nonsmokers. The relationship between fatigue resistance and SDH activity in nonsmokers (r = 0.93; p = 0.02) is absent in smokers. This indicates that the lower muscle fatigue resistance of smokers can likely be attributed to causes other than differences in oxidative metabolism and capillarization. [ABSTRACT FROM AUTHOR]

Objectives: We evaluated whether human adult bone marrow-derived mesenchymal stem cells (hMSCs) could repair an experimentally induced conduction block in cardiomyocyte cultures. Background: Autologous stem cell therapy is a novel treatment option for patients with heart disease. However, detailed electrophysiological characterization of hMSCs is still lacking. Methods: Neonatal rat cardiomyocytes were seeded on multi-electrode arrays. After 48 h, abrasion of a 200- to 450-μm–wide channel caused conduction block. Next, we applied adult hMSCs (hMSC group, n = 8), human skeletal myoblasts (myoblast group, n = 7), rat cardiac fibroblasts (fibroblast group, n = 7), or no cells (control group, n = 7) in a channel-crossing pattern. Cross-channel electrical conduction was analyzed after 24 and 48 h. Intracellular action potentials of hMSCs and cardiomyocytes were recorded. Immunostaining for connexins and intercellular dye transfer (calcein) assessed the presence of functional gap junctions. Results: After creation of conduction block, two asynchronously beating fields of cardiomyocytes were present. Application of hMSCs restored synchronization between the two fields in five of eight cultures after 24 h. Conduction velocity across hMSCs (0.9 ± 0.4 cm/s) was approximately 11-fold slower than across cardiomyocytes (10.4 ± 5.8 cm/s). No resynchronization occurred in the myoblast, fibroblast, or control group. Intracellular action potential recordings indicated that conduction across the channel presumably occurred by electrotonic impulse propagation. Connexin-43 was present along regions of hMSC-to-cardiomyocyte contact, but not along regions of cardiomyocyte-to-myoblast or cardiomyocyte-to-fibroblast contact. Calcein transfer from cardiomyocytes to hMSCs was observed within 24 h after co-culture initiation. Conclusions: Human mesenchymal stem cells are able to repair conduction block in cardiomyocyte cultures, probably through connexin-mediated coupling. [Copyright &y& Elsevier]

Background: Raman spectroscopy has the unique potential to detect and quantify cholesterol and calcification in an atherosclerotic plaque in vivo.Objective: To evaluate the sensitivity and specificity of this technique for detecting cholesterol or calcification in human coronary artery and aorta specimens ex vivo, using a compact clinical fibreoptic based Raman system developed for in vivo applications.Design: From nine coronary arteries and four aorta specimens, 114 sites were evaluated for the presence of cholesterol and calcification by Raman spectroscopy and standard histology. Raman spectra were acquired and evaluated on-line in around five seconds.Results: The correlation between Raman spectroscopy and histology was r = 0.68 for cholesterol and r = 0.71 calcification in the plaque (p < 0.0001). Sensitivity and specificity for detecting cholesterol and calcification were excellent: receiver operating characteristic (ROC) analysis for each of the components revealed areas under the curves of > 0.92 (p < 0.0001). At the optimal cut-off values determined by ROC analysis, positive predictive values of > 80% and negative predictive values of > 90% were obtained.Conclusions: On-line real time catheter based Raman spectroscopy detects accumulation of cholesterol and calcification in atherosclerotic plaque with high sensitivity and specificity. [ABSTRACT FROM AUTHOR]

Background: Oxidation susceptibility of lipids in vitro is considered to reflect the exposure of lipids to oxidation stress in vivo which is related to cardiovascular morbidity. This study examined the effect of pravastatin therapy on serum oxidation susceptibility, particularly in relation to endothelial function of coronary arteries. Methods: The participants were recruited from the Pravastatin-Related Effects Following Angioplasty on Coronary Endothelium trial, a double-blinded, placebo-controlled, randomized, multi-center study designed to analyze the effect of parvastatin treatment on endothelial function in previously dilated and normal coronary arteries. Serial, graded, intra-coronary acetylcholine infusions were used to assess endothelial function. In vitro, copper-induced, serum oxidation parameters were determined at randomization and at time of coronary endothelial function assessment. Results: Oxidation parameters were determined in 45 patients (pravastatin 23, placebo 22). Pravastatin therapy significantly improved serum oxidation lag time (+8%, P<0.05), maximal diene formation rate (−22%, P<0.01) and total amount of dienes formed after 5 h (−16%, P<0.01). These parameters remained essentially unchanged in the placebo group. Acetylcholine-evoked responses were positively correlated to therapy-induced change in serum oxidation susceptibility in the dilated segment group (r2=0.56, P=0.006). Conclusion: Pravastatin''s beneficial effect on endothelial dysfunction of dilated coronary segments may be secondary to pravastatin''s improvement of oxidation susceptibility. [Copyright &y& Elsevier]

We have examined whether melanin affects Ca2+ homeostasis in cultured normal human melanocytes. Intracellular Ca2+ concentrations ([Ca2+ ]i ), were measured in four Caucasian and in three Negroid melanocyte cultures. Under resting conditions [Ca2+ ]i was around 100 nM in all cultures, but differences between cells within cultures were observed. All cultures responded to endothelin-1 (ET-1) with increases in [Ca2+ ]i and there were no differences between Caucasian and Negroid cultures. However, large differences in responses between cells within cultures were observed, indicating that melanocyte cultures are very heterogeneous. The addition of 2.5 mM CaCl2 to melanocytes kept in Ca2+ -free medium resulted in rapid and transient increases in [Ca2+ ]i of up to 1500 nM. These increases were on average more than two times smaller in melanocyte cultures established from Negroid donors compared with Caucasian cultures. In addition, well melanized Caucasian melanocytes, cultured in the presence of 400 μM tyrosine and 10 mM NH4 Cl, showed a reduced increase in cytoplasmic Ca2+ concentration upon the addition of extracellular Ca2+ . The difference in maintaining Ca2+ homeostasis between poorly and well melanized melanocytes may be the result of the clearance of cytoplasmic Ca2+ into melanosomes and the greater capacity for this in the more pigmented melanocytes. [ABSTRACT FROM AUTHOR]

Introduction: Hypercholesterolemia is a well-known risk factor for developing atherosclerosis and subsequently for the risk of a myocardial infarction (MI). Moreover, it might also be related to the extent of damaged myocardium in the event of a MI. The aim of this study was to evaluate the association of baseline low density lipoprotein-cholesterol (LDL-c) level with infarct size in patients with ST-segment elevation myocardial infarction (STEMI) after primary percutaneously coronary intervention (pPCI). Methods: Baseline blood samples were obtained from all patients admitted between 2004 and 2014 with STEMI who underwent pPCI. Patients were excluded in case of out of hospital cardiac arrest, treatment delay of at least 10 h or no complete reperfusion after pPCI in the culprit vessel. Peak creatine kinase (CK) level was used for infarct size estimation, defined as the maximal value during admission. Results: A total of 2248 patients were included in this study (mean age 61.8 ± 12.2 years; 25.0% female). Mean LDL-c level was 3.6 ± 1.1 mmol/L and median peak CK level was 1275 U/L (IQR 564–2590 U/L). Baseline LDL-c level [β = 0.041; (95% CI 0.019–0.062); p < 0.001] was independently associated with peak CK level. Furthermore, left anterior descending artery as culprit vessel, initial TIMI 0–1 flow in the culprit vessel, male gender, and treatment delay were also correlated with high peak CK level (p < 0.05). Prior aspirin therapy was associated with lower peak CK level [β = − 0.073 (95% CI − 0.146 to 0.000), p = 0.050]. Conclusion: This study demonstrates that besides the more established predictors of infarct size, elevated LDL-c is associated with augmented infarct size in patients with STEMI treated with pPCI. [ABSTRACT FROM AUTHOR]

Uncoupling of mitochondrial proton pumping and adenosine triphosphate (ATP) production lowers mitochondrial efficiency. Current methods to determine mitochondrial efficiency require substantial amounts of tissue and permeabilization or isolation procedures. A simple histochemical method has been described by Meijer and Vloedman (Histochemistry 69:217-232, 1980, 10.1007/BF00489769), but this was not quantitative. We found linear correlations between (1) absorbance and sections thickness and (2) absorbance and incubation time. Because the method obeys Lambert-Beer's law, we can estimate ATP/O2 ratios for healthy and overloaded right-sided rat myocardium. We related mitochondrial efficiency to the ratio between cardiolipin and its precursor phosphatidylglycerol. We found a non-linear relationship between mitochondrial efficiency and this ratio, indicating that lower mitochondrial efficiency as found in experimental pulmonary hypertension may be due to altered composition of the mitochondrial inner membrane. We conclude that the histochemical method of Meijer and Vloedman can be applied to quantify mitochondrial efficiency. [ABSTRACT FROM AUTHOR]

Background: Antithrombin (AT) is a critical regulator of coagulation, and its overall activity is typically measured using functional tests. A large number of molecular forms of AT have been identified and each individual carries multiple molecular proteoforms representing variable activities. Conventional functional tests are completely blind for these proteoforms. A method that ensures properly defined measurands for AT is therefore needed. We here assess whether mass spectrometry technology, in particular multiple reaction monitoring (MRM), is suitable for the quantification of AT and the qualitative detection of its molecular proteoforms. Methods: Plasma proteins were denatured, reduced and alkylated prior to enzymatic digestion. MRM transitions were developed towards tryptic peptides and glycopeptides using AT purified from human plasma. For each peptide, three transitions were measured, and stable isotope-labeled peptides were used for quantitation. Completeness of digestion was assessed using digestion time curves. Results: MRM transitions were developed for 19 tryptic peptides and 4 glycopeptides. Two peptides, FDTISEK and FATTFYQHLADSK, were used for quantitation, and using a calibration curve of isolated AT in 40 g/L human serum albumin, CVs below 3.5% were obtained for FDTISEK, whereas CVs below 8% were obtained for FATTFYQHLADSK. Of the 26 important AT mutations, 20 can be identified using this method, while altered glycosylation profiles can also be detected. Conclusions: We here show the feasibility of the liquid chromatography multiple reaction monitoring mass spectrometry (LC-MRM-MS) technique for the quantitation of AT and the qualitative analysis of most of its molecular proteoforms. Knowing the measurands will enable standardization of AT tests by providing in-depth information on the molecular proteoforms of AT. [ABSTRACT FROM AUTHOR]

Rowers need to combine high sprint and endurance capacities. Muscle morphology largely explains muscle power generating capacity, however, little is known on how muscle morphology relates to rowing performance measures. The aim was to determine how muscle morphology of the vastus lateralis relates to rowing ergometer performance, sprint and endurance capacity of Olympic rowers. Eighteen rowers (12♂, 6♀, who competed at 2016 Olympics) performed an incremental rowing test to obtain maximal oxygen consumption, reflecting endurance capacity. Sprint capacity was assessed by Wingate cycling peak power. M. vastus lateralis morphology (volume, physiological cross-sectional area, fascicle length and pennation angle) was derived from 3-dimensional ultrasound imaging. Thirteen rowers (7♂, 6♀) completed a 2000-m rowing ergometer time trial. Muscle volume largely explained variance in 2000-m rowing performance (R2 = 0.85), maximal oxygen consumption (R2 = 0.65), and Wingate peak power (R2 = 0.82). When normalized for differences in body size, maximal oxygen consumption and Wingate peak power were negatively related in males (r = −0.94). Fascicle length, not physiological cross-sectional area, attributed to normalized peak power. In conclusion, vastus lateralis volume largely explains variance in rowing ergometer performance, sprint and endurance capacity. For a high normalized sprint capacity, athletes may benefit from long fascicles rather than a large physiological cross-sectional area. [ABSTRACT FROM AUTHOR]

Chronic hypoxia is associated with muscle wasting and decreased oxidative capacity. By contrast, training under hypoxia may enhance hypertrophy and increase oxidative capacity as well as oxygen transport to the mitochondria, by increasing myoglobin (Mb) expression. The latter may be a feasible strategy to prevent atrophy under hypoxia and enhance an eventual hypertrophic response to anabolic stimulation. Mb expression may be further enhanced by lipid supplementation. We investigated individual and combined effects of hypoxia, insulin-like growth factor (IGF)-1 and lipids, in mouse skeletal muscle C2C12 myotubes. Differentiated C2C12 myotubes were cultured for 24 h under 20%, 5% and 2% oxygen with or without IGF-1 and/or lipid treatment. In culture under 20% oxygen, IGF-1 induced 51% hypertrophy. Hypertrophy was only 32% under 5% and abrogated under 2% oxygen. This was not explained by changes in expression of genes involved in contractile protein synthesis or degradation, suggesting a reduced rate of translation rather than of transcription. Myoglobin mRNA expression increased by 75% under 5% O2 but decreased by 50% upon IGF-1 treatment under 20% O2, compared to control. Inhibition of mammalian target of rapamycin (mTOR) activation using rapamycin restored Mb mRNA expression to control levels. Lipid supplementation had no effect on Mb gene expression. Thus, IGF-1-induced anabolic signaling can be a strategy to improve muscle size under mild hypoxia, but lowers Mb gene expression. [ABSTRACT FROM AUTHOR]

A major problem in chronic heart failure is the inability of hypertrophied cardiomyocytes to maintain the required power output. A Hill-type oxygen diffusion model predicts that oxygen supply is limiting in hypertrophied cardiomyocytes at maximal rates of oxygen consumption and that this limitation can be reduced by increasing the myoglobin (Mb) concentration. We explored how cardiac hypertrophy, oxidative capacity, and Mb expression in right ventricular cardiomyocytes are regulated at the transcriptional and translational levels in an early stage of experimental pulmonary hypertension, in order to identify targets to improve the oxygen supply/demand ratio. Male Wistar rats were injected with monocrotaline to induce pulmonary hypertension (PH) and right ventricular heart failure. The messenger RNA (mRNA) expression levels per nucleus of growth factors insulin-like growth factor-1Ea (IGF-1Ea) and mechano growth factor (MGF) were higher in PH than in healthy controls, consistent with a doubling in cardiomyocyte cross-sectional area (CSA). Succinate dehydrogenase (SDH) activity was unaltered, indicating that oxidative capacity per cell increased. Although the Mb protein concentration was unchanged, Mb mRNA concentration was reduced. However, total RNA per nucleus was about threefold higher in PH rats versus controls, and Mb mRNA content expressed per nucleus was similar in the two groups. The increase in oxidative capacity without an increase in oxygen supply via Mb-facilitated diffusion caused a doubling of the critical extracellular oxygen tension required to prevent hypoxia (PO). We conclude that Mb mRNA expression is not increased during pressure overload-induced right ventricular hypertrophy and that the increase in myoglobin content per myocyte is likely due to increased translation. We conclude that increasing Mb mRNA expression may be beneficial in the treatment of experimental PH. [ABSTRACT FROM AUTHOR]

Sodium alginate is an effective biomaterial for tissue engineering applications. Non-purified alginate is contaminated with protein, lipopolysaccharide, DNA, and RNA, which could elicit adverse immunological reactions. We developed a purification protocol to generate biocompatible alginate based on (a) activated charcoal treatment, (b) use of hydrophobic membrane filtration (we used hydrophobic polyvinylidene difluoride membranes to remove organic contaminants), (c) dialysis, and finally (d) ethanol precipitation. Using this approach, we could omit pre-treatment with chloroform and significantly reduce the quantities of reagents used. Purification resulted in reduction of residual protein by 70% down to 0.315 mg/g, DNA by 62% down to 1.28 µg/g, and RNA by 61% down to less than 10 µg/g, respectively. Lipopolysaccharide levels were reduced by >90% to less than 125 EU/g. Purified alginate did not induce splenocyte proliferation in vitro. Three-dimensional scaffolds generated from purified alginate did not elicit a significant foreign body reaction, fibrotic overgrowth, or macrophage infiltration 4 weeks after implantation. This study describes a simplified and economical alginate purification method that results in alginate purity, which meets clinically useful criteria. [ABSTRACT FROM AUTHOR]

Background: Near-infrared spectroscopy (NIRS) measurements of oxygenation reflect O2 delivery and utilization in exercising muscle and may improve detection of a critical exercise threshold. Purpose: First, to detect an oxygenation breakpoint (Δ[O2HbMb-HHbMb]-BP) and compare this breakpoint to ventilatory thresholds during a maximal incremental test across sexes and training status. Second, to assess reproducibility of NIRS signals and exercise thresholds and investigate confounding effects of adipose tissue thickness on NIRS measurements. Methods: Forty subjects (10 trained male cyclists, 10 trained female cyclists, 11 endurance trained males and 9 recreationally trained males) performed maximal incremental cycling exercise to determine Δ[O2HbMb-HHbMb]-BP and ventilatory thresholds (VT1 and VT2). Muscle haemoglobin and myoglobin O2 oxygenation ([HHbMb], [O2HbMb], SmO2) was determined in m. vastus lateralis. Δ[O2HbMb-HHbMb]-BP was determined by double linear regression. Trained cyclists performed the maximal incremental test twice to assess reproducibility. Adipose tissue thickness (ATT) was determined by skinfold measurements. Results: Δ[O2HbMb-HHbMb]-BP was not different from VT1, but only moderately related (r = 0.58–0.63, p<0.001). VT1 was different across sexes and training status, whereas Δ[O2HbMb-HHbMb]-BP differed only across sexes. Reproducibility was high for SmO2 (ICC = 0.69–0.97), Δ[O2HbMb-HHbMb]-BP (ICC = 0.80–0.88) and ventilatory thresholds (ICC = 0.96–0.99). SmO2 at peak exercise and at occlusion were strongly related to adipose tissue thickness (r2 = 0.81, p<0.001; r2 = 0.79, p<0.001). Moreover, ATT was related to asymmetric changes in Δ[HHbMb] and Δ[O2HbMb] during incremental exercise (r = -0.64, p<0.001) and during occlusion (r = -0.50, p<0.05). Conclusion: Although the oxygenation threshold is reproducible and potentially a suitable exercise threshold, VT1 discriminates better across sexes and training status during maximal stepwise incremental exercise. Continuous-wave NIRS measurements are reproducible, but strongly affected by adipose tissue thickness. [ABSTRACT FROM AUTHOR]

An abstract of the study "Body weight changes explained by lung weight and myocardial HIF 1 alpha expression in monocrotaline-treated rats," by B. J. Van Beek-Harmsen et al, is presented.

Background: Serum troponin assays, widely used to detect acute cardiac ischemia, might be useful biomarkers to detect chronic cardiovascular disease (CVD). Cardiac-specific troponin-I (cTnI) and troponin-T (cTnT) generally detect myocardial necrosis equally well. In dialysis patients however, serum cTnT levels are often elevated, unlike cTnI levels. The present study aims to elucidate the associations of cTnI and cTnT with CVD in clinically stable dialysis patients. Methods: Troponin levels were measured using 5th generation hs-cTnT assays (Roche) and STAT hs-cTnI assays (Abbott) in a cohort of dialysis patients. Serum troponin levels were divided into tertiles with the lowest tertile as a reference value. Serum troponins were associated with indicators of CVD such as left ventricular mass index (LVMI), left ventricular ejection fraction (LVEF) and the presence of coronary artery disease (CAD). Associations were explored using regression analysis. Results: We included 154 consecutive patients, 68±7 years old, 77% male, 70% hemodialysis. Median serum cTnT was 51ng/L (exceeding the 99th percentile of the healthy population in 98%) and median serum cTnI was 13ng/L (elevated in 20%). A high cTnI (T3) was significantly associated with a higher LVMI (Beta 31.60; p=0.001) and LVEF (Beta -4.78; p=0.005) after adjusting for confounders whereas a high serum cTnT was not. CAD was significantly associated with a high cTnT (OR 4.70 p=0.02) but not with a high cTnI. Unlike cTnI, cTnT was associated with residual renal function (Beta:-0.09; p=0.006). Conclusion: In the present cohort, serum cTnI levels showed a stronger association with LVMI and LVEF than cTnT. However, cTnT was significantly associated with CAD and residual renal function, unlike cTnI. Therefore, cTnI seems to be superior to cTnT as a marker of left ventricular dysfunction in asymptomatic dialysis patients, while cTnT might be better suited to detect CAD in these patients. [ABSTRACT FROM AUTHOR]

Background: The leading cause of mortality due to pulmonary arterial hypertension (PAH) is failure of the cardiac right ventricle. It has long been hypothesized that during the development of chronic cardiac failure the heart becomes energy deprived, possibly due to shortage of oxygen at the level of cardiomyocyte mitochondria. However, direct evaluation of oxygen tension levels within the in vivo right ventricle during PAH is currently lacking. Here we directly evaluated this hypothesis by using a recently reported technique of oxygen-dependent quenching of delayed fluorescence of mitochondrial protoprophyrin IX, to determine the distribution of mitochondrial oxygen tension (mitoPO2) within the right ventricle (RV) subjected to progressive PAH. Methods: PAH was induced through a single injection of monocrotaline (MCT). Control (saline-injected), compensated RV hypertrophy (30 mg/kg MCT; MCT30), and RV failure (60 mg/kg MCT; MCT60) rats were compared 4 wk after treatment. The distribution of mitoPO2 within the RV was determined in mechanically-ventilated, anaesthetized animals, applying different inspired oxygen (FiO2) levels and two increment dosages of dobutamine. Results: MCT60 resulted in RV failure (increased mortality, weight loss, increased lung weight), MCT30 resulted in compensated RV hypertrophy. At 30% or 40% FiO2, necessary to obtain physiological arterial PO2 in the diseased animals, RV failure rats had significantly less mitochondria (15% of total mitochondria) in the 0-20 mmHg mitoPO2 range than hypertrophied RV rats (48%) or control rats (54%). Only when oxygen supply was reduced to 21% FiO2, resulting in low arterial PO2 for the MCT60 animals, or when oxygen demand increased with high dose dobutamine, the number of failing RV mitochondria with low oxygen became similar to control RV. In addition, metabolic enzyme analysis revealed similar mitochondrial mass, increased glycolytic hexokinase activity following MCT, with increased lactate dehydrogenase activity only in compensated hypertrophied RV. Conclusions: Our novel observation of increased mitochondrial oxygenation suggests down-regulation of in vivo mitochondrial oxygen consumption, in the absence of hypoxia, with transition towards right ventricular failure induced by pulmonary arterial hypertension. [ABSTRACT FROM AUTHOR]

Background In pulsed-dose rate prostate brachytherapy the dose is delivered during 48 hours after implantation, making the treatment sensitive to oedematic effects possibly affecting dose delivery. The aim was to study changes in prostate volume during treatment by analysing catheter configurations on three subsequent scans. Methods Prostate expansion was determined for 19 patients from the change in spatial distribution of the implanted catheters, using three CT-scans: a planning CT (CT1) and two CTs after 24 and 48 hours (CT2, CT3). An additional 4 patients only received one repeat CT (after 24 hours). The mean radial distance (MRD) of all dwell positions to the geometric centre of all dwell positions used was calculated to evaluate volume changes. From three implanted markers changes in inter-marker distances were assessed. The relative shifts of all dwell positions were determined using catheter- and marker-based registrations. Wilcoxon signed-rank tests were performed to compare the results from the different time points. The MRDs measured on the two repeat CTs were significantly different from CT1. The mean prostate volume change derived from the difference in MRD was +4.3% (range −9.3% to +15.6%) for CT1-CT2 (p < .05) and +4.4% (range −7.5% to +16.3%) for CT1-CT3 (p < .05). These values represented a mean increase of 1.2 cm3 in the first 24 hours and 1.5 cm3 in the subsequent 24 hours. There was no clear sign of prostate expansion from the change in intermarker distance (CT1-CT2: 0.2 ± 1.8 mm; CT1-CT3: 0.6 ± 2.2 mm). Catheter configuration remained stable; shifts in catheter positions were largest in the C-C direction: 0 ± 1.8 mm for CT1-CT2 and 0 ± 1.4 mm for CT2-CT3. Conclusions The volume changes derived from catheter displacements were small and therefore considered clinically insignificant. Implant configuration remains stable during 2 days of treatment, confirming the safety of this technique. [ABSTRACT FROM AUTHOR]

In this study, we have followed up on previous liquid chromatography (LC) multiple reaction monitoring (MRM) mass spectrometry (MS) approaches for measurement of apolipoprotein (apo) A-I and apo B100 in serum aiming for implementation of a multiplexed assay in a clinical chemistry laboratory with full metrological traceability. Signature peptides were selected and detected by dynamic MRM, and stable isotope labeled (SIL)-peptides were used as internal standards. Five apo A-I and four apo B100 peptides were measured in serum digests with linearity (R 2 > 0.992) in the physiologically relevant concentration ranges. Linearity with regard to protein concentration was ascertained at five concentration levels (R 2 > 0.926 and R 2 > 0.965, for the apo A-I and apo B100 peptides, respectively). Three native value-assigned sera were used as external calibrators for further method verification. Imprecision values on sample preparation and LC-MS/MS acquisition were below the established minimal specifications for apo A-I and apo B100 (5.0% and 5.3%, respectively). Correlation of LC-MS/MS results with immunoturbidimetric assay results, for normo- and hypertriglyceridemic samples, showed R 2 > 0.944 for apo A-I and R 2 > 0.964 for apo B100. This LC-MS/MS method has potential for clinical application in normo- and dyslipidemic patients. Biological significance Measurement of apo A-I and apo B100 may offer an alternative to high and low density lipoprotein cholesterol (HDL-c and LDL-c) methods for cardiovascular disease risk assessment in dyslipidemic patients [1]. An LC-MS/MS method for apo A-I and apo B100 has the advantage of antibody independent and specific detection of protein signature peptides. The introduction of an LC-MS/MS method for apo A-I and apo B100 can serve as an example for many existing and newly developed (multiplex) biomarker methods in quantitative clinical chemistry proteomics (qCCP). Such LC-MS/MS methods should meet basic clinical chemistry principles with regard to test evaluation [2] . Criteria for imprecision should be pre-defined, e.g., based on biological variation. The use of commutable and traceable serum-based calibrators will improve inter-laboratory reproducibility of LC-MS/MS methods and may contribute to a more rapid transition of biomarker discovery to clinical utility with benefit for the patient treatment and improvement of general health care. [ABSTRACT FROM AUTHOR]