A new approach to quantify visceral fat via bioelectrical impedance analysis and ultrasound compared to MRI.

BACKGROUND: Visceral adipose tissue (VAT) has been linked to systemic proinflammatory characteristics, and measuring it accurately usually requires sophisticated instruments. This study aimed to estimate VAT applying a simpler method that uses total subcutaneous fat and total body fat (BF) measurements. METHOD: As part of our experimental approach, the subcutaneous fat mass (SFT) was measured via US (SFTtotal), and VAT was quantified by assessing MRI data. Both parameters were added to obtain total body fat (BFcalc). Those results were then compared to values obtained from a bioelectrical impedance analysis (BFBIA). Multiple regression analyses were employed to develop a simplified sex-specific equation for SFT, which was subsequently used in conjunction with BFBIA to determine VAT (VATEq). RESULT: We observed excellent reliability between BFBIA and BFcalc, with no significant difference in body fat values (20.98 ± 8.36 kg vs. 21.08 ± 8.81 kg, p = 0.798, ICC 0.948). VATEq_female/male revealed excellent reliability when compared to VATMRI, and no significant difference appeared (women: 0.03 ± 0.66 kg with a 95% CI ranging from -1.26 kg to 1.32 kg, p = 0.815, ICC: 0.955.; men: -0.01 ± 0.85 kg with a 95% CI ranging from -1.69 kg to 1.66 kg, p = 0.925, ICC: 0.952). CONCLUSION: Taking an experimental approach, VAT can be determined without MRI.

C-kitpos cells in the human left atrial appendage.

Background: Subpopulations of myocardial c-kitpos cells have the ability to stimulate regeneration in ischemic heart disease by paracrine effects. The left atrial appendage (LAA), which is easy accessible during cardiac surgery, may represent a perfect source for c-kitpos cell extraction for autologous cell therapies in the living human. So far, frequency and distribution of c-kitpos cells in LAA are unknown. Methods: LAAs of patients who underwent cardiac surgery due to coronary artery disease (coronary artery bypass graft, CABG), valvular heart disease or both and of two body donors were examined. Tissue was fixed in 4 % paraformaldehyde, embedded in paraffin, dissected in consecutive sections and stained for c-kitpos cells. In parallel, grade of fibrosis, amount of fat per section and cells positive for mast cell tryptase were examined. Results: We collected 27 LAAs (37.0 % female, mean left ventricular ejection fraction 50.4 %, 63.0 % persistent atrial fibrillation (AF)). Most of the patients underwent combined CABG and valve surgery (51.9 %). C-kitpos cells were detected in 3 different regions: A) Attached to the epicardial fat layer, B) close to vascular structures and C) between cardiomyocytes. C-kitpos cells ranged from 0.05 c-kitpos cells per mm2 to 67.5 c-kitpos cells per mm2. We found no association between number of c-kitpos cells and type of AF, amount of fibrosis or amount of fat. Up to 72 % of c-kitpos cells also showed a positive staining for mast cell tryptase. Conclusion: C-kitpos cells are frequent in LAAs of cardiovascular patients with a rather homogenous distribution throughout the LAA. The LAA can therefore be considered as a source for extraction of a reasonable quantity of autologous cardiac progenitor cells in the living human patient.

Measurement of subcutaneous fat tissue: reliability and comparison of caliper and ultrasound via systematic body mapping.

Caliper and ultrasound (US) are used to measure subcutaneous fat tissue depth (SFT) and then to calculate total body fat. There is no evidence-based recommendation as to whether caliper or US are equally accurate. The aim of this paper was therefore to compare reliability of both methods. In this methodical study, 54 participants (BMI: 24.8 ± 3.5 kg/m2; Age: 43.2 ± 21.7 years) were included. Using systematic body mapping, the SFT of 56 areas was measured. We also analyzed 4 body sites via MRI. A comparison between caliper and US detected clear differences in mean SFT of all areas (0.83 ± 0.33 cm vs. 1.14 ± 0.54 cm; p < 0.001) showing moderate reliability (ICC 0.669, 95%CI: 0.625-0.712). US and MRI revealed in the abdominal area a SFT twice as thick as caliper (2.43 ± 1.36 cm vs. 2.26 ± 1.32 cm vs. 1.15 ± 0.66 cm; respectively). Caliper and US revealed excellent intrarater (ICC caliper: 0.944, 95%CI: 0.926-0.963; US: 0.934, 95%CI: 0.924-0.944) and good interrater reliability (ICC caliper: 0.794, 95%CI: 0.754-0.835; US: 0.825, 95%CI: 0.794-0.857). Despite the high reliability in measuring SFT that caliper and US show, our comparison of the two methods yielded clear differences in SFT, particularly in the abdominal area. In accuracy terms, US is preferable for most mapping areas.

Outcomes following cardiac sympathetic denervation in patients with structural heart disease and refractory ventricular arrhythmia.

AIM: Cardiac sympathetic denervation (CSD) has been introduced as a bailout therapy in patients with structural heart disease and refractory ventricular arrhythmias (VAs), but available data are scarce. Purpose of this study was to estimate immediate results, complications, and mid-term outcomes of CSD following recurrent VA after catheter ablation. METHODS AND RESULTS: Adult patients who underwent CSD in the Heart Center Leipzig from March 2017 to February 2021 were retrospectively analysed. Follow-up (FU) was executed via implantable cardioverter defibrillator (ICD) interrogation, telephone interviews, and reviewing medical records. Twenty-one patients (age 63.7 ± 14.4 years, all men, 71.4% non-ischaemic cardiomyopathy, left ventricular ejection fraction 31.6 ± 12.6%) received CSD via video-assisted thoracoscopic surgery (90.5% bilateral, 9.5% left-sided only). Indication for CSD was monomorphic ventricular tachycardia in 76.2% and ventricular fibrillation in 23.8 with 71.4% of patients presenting with electrical storm before index hospitalization. Procedure-related major complications occurred in 9.5% of patients. In-hospital adverse events not related to surgery were common (28.6%) and two patients died during the index hospital stay. During FU (mean duration 9.1 ± 6.5 months), five more patients died. Of the remaining patients, 38.5 and 76.9% were free from any VA or ICD shocks, respectively. CONCLUSIONS: The CSD showed additional moderate efficacy to suppress VAs, when performed as a bailout therapy after previously unsuccessful catheter ablation. At 9 months, it was associated with freedom of ICD shocks in two-thirds of patients. In a population with many comorbidities, the rate of CSD-related complications was acceptable, although there was an overall high risk of procedure unrelated adverse events and death.

Decreased exercise capacity in young athletes using self-adapted mouthguards.

PURPOSE: There is evidence of both the preventive effects and poor acceptance of mouthguards. There are various effects on performance depending on the type of mouthguard model. Hemodynamic responses to wearing a mouthguard have not been described. The aim of this study was to investigate the effects of self-adapted mouthguards with breathing channels (SAMGvent). METHODS: In this randomized crossover study, 17 healthy, active subjects (age 25.12 ± 2.19 years) underwent body plethysmography and performed two incremental exertion tests wearing a (SAMGvent) and not wearing (CON) a mouthguard. Blood lactate, spirometrics, and thoracic impedance were measured during these maximum exercise tests. RESULTS: The mean values using a SAMGvent revealed significantly greater airway resistance compared to CON (0.53 ± 0.16 kPa·L-1 vs. 0.35 ± 0.10 kPa·L-1, respectively; p = < 0.01). At maximum load, ventilation with SAMGvent was less than CON (118.4 ± 28.17 L min-1 vs. 128.2 ± 32.16 L min-1, respectively; p = < 0.01). At submaximal loads, blood lactate responses with SAMGvent were higher than CON (8.68 ± 2.20 mmol·L-1 vs. 7.89 ± 1.65 mmol·L-1, respectively; p < 0.01). Maximum performance with a SAMGvent was 265.9 ± 59.9 W, and without a mouthguard was 272.9 ± 60.8 W (p < 0.01). Maximum stroke volume was higher using a SAMGvent than without using a mouthguard (138.4 ± 29.9 mL vs. 130.2 ± 21.2 mL, respectively; p < 0.01). CONCLUSION: Use of a self-adapted mouthguard led to increased metabolic effort and a significant reduction in ventilation parameters. Unchanged oxygen uptake may be the result of cardiopulmonary compensation and increased breathing efforts, which slightly affects performance. These results and the obvious preventive effects of mouthguards support their use in sports.

Effects of Custom-made Mouthguards on Cardiopulmonary Exercise Capacity.

The importance of using mouthguards as well as their low acceptance rate have been demonstrated. The aim of this study was to investigate the influence of customized mouthguards on hemodynamics.. This randomized crossover study used data from 13 subjects (23.5±1.4 years). The cardiopulmonary and metabolic parameters were observed during ergometer tests without mouthguard (control) in comparison to two types of mouthguards (with and normal without breathing channels). Maximum ventilation was significantly decreased with the normal mouthguard (113.3±30.00 l ∙ min-1) in contrast to the mouthguard with breathing channels (122.5±22.9 l ∙ min-1) and control (121.9±30.8 l ∙ min-1). Also the inspiration time was longer when using the normal mouthguard (0.70±0.11 s) compared to the mouthguard with breathing channels (0.63±0.11 s) and control (Co 0.64±0.10 s). Lactate was also increased under the influence of the mouthguard with breathing channels (10.72±1.4 mmol ∙ l-1) compared to the control (9.40±1.77 mmol ∙ l-1) and the normal mouthguard (9.02±1.67 mmol ∙ l-1). In addition, stroke volume kinetics (p=0.048) and maximum heart rates (p=0.01) show changes. Despite equal levels of oxygen uptake and performances under all three conditions, the use of mouthguards showed differences in cardiopulmonary parameters. The use of mouthguards during exercise does not affect physical performance and can be recommended for injury prevention.

[Verbesserung der Lebensqualität durch integrative onkologische Rehabilitation]. / Sustained Improvement of Quality of Life following Integrative Oncological Rehabilitation.

Hintergrund: Die onkologische Rehabilitation ist integraler Bestandteil der Versorgung krebskranker Menschen. Nach einer dreiwöchigen stationären Rehabilitation mit multimodalem und integrativem Ansatz wurden die Effekte auf Belastungen und Lebensqualität der Patienten überprüft. Patienten und Methoden: 74 Krebspatienten erhielten ein komplexes Therapieprogramm, das Therapien zur Verbesserung der funktionalen Gesundheit, zur Reduktion psychosozialer Belastungen und komplementäre Massnahmen beinhaltete. Der Erfolg der Therapie wurde mit validierten Fragebögen am Abschluss der Rehabilitation (T2) und 3 Monate danach (T3) bestimmt. Ergebnisse: Es zeigte sich eine signifikante Besserung von Distress, Angst, Depression, Fatigue und Lebensqualitätsfunktionsskalen zum Zeitpunkt T2 und T3. Von T2 nach T3 war der Therapieeffekt rückläufig, ohne die Werte von T1 zu erreichen. Schlussfolgerungen: Eine multimodale, integrative onkologische Rehabilitation führt zu einer über 3 Monate anhaltenden Besserung des subjektiven Befindens der Patienten. Dieses Therapiekonzept sollte in einer Folgestudie mit einer Standardrehabilitation verglichen werden. BACKGROUND: Oncological rehabilitation is an integral part in the care of cancer patients. Following an inpatient rehabilitation of 3 weeks' duration with multidimensional and integrative components, the effects on distress and quality of life were measured. PATIENTS AND METHODS: 74 cancer patients received a complex treatment program, including treatments for improvement of functional health, reduction of psychosocial distress and complementary therapies. The treatment outcome was evaluated with validated questionnaires at the end of the rehabilitation (T2) and 3 months thereafter (T3). RESULTS: We observed significant improvement of distress, anxiety, depression, fatigue and quality of life at T2 and T3. In the interval from T2 to T3, the treatment effect was declining, without reaching the values of T1. CONCLUSIONS: A multidimensional integrative oncological rehabilitation improves the subjective condition of the patients over a 3-month period. This treatment concept should be tested in a comparative study against standard rehabilitation.

Isolation of Native Soluble and Membrane-Bound Protein Complexes from Yeast Saccharomyces cerevisiae.

Immunoprecipitation is a traditional approach to isolate single proteins or native protein complexes from a complex sample mixture. The original method makes use of specific antibodies against endogenous proteins or epitope tags, which are first bound to the target protein and then isolated with protein A beads. An advancement of this method is the application of a protein A tag fused to the target protein and the affinity-purification of the tagged protein with human Immunoglobulin G chemically cross-linked to a sepharose matrix. This method will be described exemplified by the purification of protein complexes of the peroxisomal membrane from yeast Saccharomyces cerevisiae.

Pex17p-dependent assembly of Pex14p/Dyn2p-subcomplexes of the peroxisomal protein import machinery.

Peroxisomal matrix protein import is facilitated by cycling receptors that recognize their cargo proteins in the cytosol by peroxisomal targeting sequences (PTS). In the following, the assembled receptor-cargo complex is targeted to the peroxisomal membrane where it docks to the docking-complex as part of the peroxisomal translocation machinery. The docking-complex is composed of Pex13p, Pex14p and in yeast also Pex17p, whose function is still elusive. In order to characterize the function of Pex17p, we compared the composition and size of peroxisomal receptor-docking complexes from wild-type and pex17Δ cells. Our data demonstrate that the deficiency of Pex17p affects the stoichiometry of the constituents of an isolated 600kDa complex and that pex17Δ cells lack a high molecular weight complex (>900kDa) of unknown function. We identified the dynein light chain protein Dyn2p as an additional core component of the Pex14p/Pex17p-complex. Both, Pex14p and Pex17p interact directly with Dyn2p, but in vivo, Pex17p turned out to be prerequisite for an association of Dyn2p with Pex14p. Finally, like pex17Δ also dyn2Δ cells lack the high molecular weight complex. As dyn2Δ cells also display reduced peroxisomal function, our data indicate that Dyn2p-dependent formation of the high molecular weight Pex14p-complex is required to maintain peroxisomal function on wild-type level.

Intra-Aortic Balloon Pump Malposition Reduces Visceral Artery Perfusion in an Acute Animal Model.

Visceral artery perfusion can be potentially affected by intra-aortic balloon pump (IABP) catheters. We utilized an animal model to quantify the acute impact of a low balloon position on mesenteric artery perfusion. In six pigs (78 ± 7 kg), a 30-cc IABP was placed in the descending aorta in a transfemoral procedure. The celiac artery (CA) and the cranial mesenteric artery (CMA) were surgically dissected. Transit time blood flow was measured for (i) baseline, (ii) 1:1 augmentation with the balloon proximal to the visceral arteries, and (iii) 1:1 augmentation with the balloon covering the visceral arteries. Blood flow in the CMA and CA was reduced by 17 and 24%, respectively, when the balloon compromised visceral arteries compared with a position above the visceral arteries (flow in mL/min: CMA: (i) 1281 ± 512, (ii) 1389 ± 287, (iii) 1064 ± 276, P < 0.05 for 3 vs. 1 and 3 vs. 2; CA: (i) 885 ± 370, (ii) 819 ± 297, (iii) 673 ± 315; P < 0.05 for 3 vs. 1). The covering of visceral arteries by an IABP balloon causes a significant reduction of visceral artery perfusion; thus, the positioning of this device during implantation is critical for obtaining a satisfactory outcome.

The role of the heart team in complicated transcatheter aortic valve implantation: a 7-year single-centre experience.

OBJECTIVES: European guidelines recommend to perform transcatheter aortic valve implantation (TAVI) within a multidisciplinary heart team. However, there is a strong drive--despite existing guidelines--to perform TAVI outside of specialized centres. The aim of this study was to clarify the necessity of on-site cardiac surgery by providing a clear insight into the complications during/after TAVI that needed surgical management. METHODS: A total of 2287 (1523 transfemoral, 752 transapical and 12 transaortic) patients, with a mean age of 84.5 ± 5.3 years, and a mean log EuroSCORE of 21.7 ± 16.3, of which 205 were female (84%), underwent TAVI since February 2006 at our institution. All procedure-related complications that required surgical interventions, whether immediate or delayed but within the initial hospital stay, were recorded and retrospectively analysed. RESULTS: Out of this cohort, 245 (10.7%) patients required surgical treatment due to major complications. A total of 42 patients (1.8%) underwent conversion to full sternotomy and 27 (1.2%) were dependent on the short-term use of the heart-lung machine. Vascular complications with surgical intervention were seen in 85 patients (3.7%), 54 patients (2.4%) had to have a rethoracotomy within their initial stay and 15 (0.7%) required a cardiac reoperation. CONCLUSIONS: Severe complications during TAVI that can only be resolved surgically will continue to occur. Therefore, each TAVI procedure should be conducted or accompanied by a cardiac surgeon and an experienced team within a specialized centre.

Synthesis and biological evaluation of urea derivatives as highly potent and selective rho kinase inhibitors.

RhoA and its downstream effector ROCK mediate stress fiber formation and cell contraction through their effects on the phosphorylation of myosin light chain (MLC). Inhibition of the RhoA/ROCK pathway has proven to be a promising strategy for several indications such as cardiovascular disease, glaucoma, and inflammatory disease. In 2010, our group reported urea-based ROCK inhibitors as potential antiglaucoma agents. These compounds showed potent IC50 values in enzymatic and cell-based assays and significant intraocular pressure (IOP)-lowering effects in rats (∼7 mmHg). (22) To develop more advanced ROCK inhibitors targeting various potential applications (such as myocardial infarction, erectile dysfunction, multiple sclerosis, etc.) in addition to glaucoma, a thorough SAR for this urea-based scaffold was studied. The detailed optimization process, counter-screening, and in vitro and in vivo DMPK studies are discussed. Potent and selective ROCK inhibitors with various in vivo pharmacokinetic properties were discovered.

Amino acid derived quinazolines as Rock/PKA inhibitors.

SAR and lead optimization studies for Rock inhibitors based on amino acid-derived quinazolines are described. Studies demonstrated that these amino acid derived quinazolinones were mainly pan-Rock (I & II) inhibitors. While selectivity against other kinases could be achieved, selectivity for most of these compounds against PKA was not achieved. This is distinct from Rock inhibitors based on non-amino acid derived quinazolinones, where high selectivity against PKA could be obtained.(22) The inhibitors presented here in some cases possessed sub-nanomolar inhibition of Rock, nanomolar potency in ppMLC cell based assays, low to fair cytochrome P-450 inhibition, and good human microsomal stability.

Outcome and incidence of re-intervention after surgical repair of tetralogy of fallot.

OBJECTIVES: Timing of primary repair of tetralogy of Fallot (TOF) remains controversial. We evaluated the long-term outcome of early primary treatment strategy in a patient cohort with TOF less than eight months of age. METHODS: A group of 120 patients with TOF less than eight months of age (5 ± 2.4 months) underwent early primary repair of TOF between October 1998 and August 2009. Sixty-one patients received a transanular (TAN) repair, and 59 patients received a right ventricular outflow tract (RVOT) + main pulmonary artery (MPA) double patch repair with concomitant pulmonary valve reconstruction. RESULT: There was no early or late mortality. The follow-up was 100% completed. There were eight reoperations and eight patients underwent catheter intervention for severe pulmonary valve insufficiency or stenosis, obstruction of right ventricular outflow tract, and stenosis of pulmonary arteries. Actuarial survival was 100% at ten years. At latest follow-up 80 patients were in NYHA Class I without any antiarrhythmic medications. On latest echocardiography, 90 (75%) patients had mild to moderate pulmonary regurgitation, and 10 had a right ventricular outflow tract gradient more than 40 mmHg. CONCLUSIONS: These data strongly support the concept of early primary repair of TOF in patients with well developed pulmonary arteries. Early primary repair is associated with an excellent early and late outcomes, an acceptable risk of reoperation and re-intervention, and a low incidence of significant right ventricular dysfunction.

Discovery and optimization of indole and 7-azaindoles as Rho kinase (ROCK) inhibitors (part-II).

Therapeutic interventions with Rho kinase (ROCK) inhibitors may effectively treat several disorders such as hypertension, stroke, cancer, and glaucoma. Herein we disclose the optimization and biological evaluation of potent novel ROCK inhibitors based on substituted indole and 7-azaindole core scaffolds. Substitutions on the indole C3 position and on the indole NH and/or amide NH positions all yielded potent and selective ROCK inhibitors (25, 42, and 50). Improvement of aqueous solubility and tailoring of in vitro and in vivo DMPK properties could be achieved through these substitutions.

Discovery and optimization of indoles and 7-azaindoles as Rho kinase (ROCK) inhibitors (part-I).

Rho kinase (ROCK) inhibitors are potential therapeutic agents to treat disorders such as hypertension, multiple sclerosis, cancers, and glaucoma. Here, we disclose the synthesis, optimization, biological evaluation of potent indole and 7-azaindole based ROCK inhibitors that have high potency on ROCK (IC(50)=1 nM) with 740-fold selectivity over PKA (47). Moreover, 47 showed very good DMPK properties making it a good candidate for further development. Finally, docking studies with a homology model of ROCK-II were performed to rationalize the binding mode of these compounds and showed the compounds bound in both orientations to take advantage to H-bonds with Lys-121 of ROCK-II.

Synthesis and biological evaluation of 4-quinazolinones as Rho kinase inhibitors.

Rho kinase (ROCK) is an attractive therapeutic target for various diseases including glaucoma, hypertension, and spinal cord injury. Herein, we report the development of a series of ROCK-II inhibitors based on 4-quinazolinone and quinazoline scaffolds. SAR studies at three positions of the quinazoline core led to the identification of analogs with high potency against ROCK-II and good selectivity over protein kinase A (PKA).

Tetrahydroisoquinoline derivatives as highly selective and potent Rho kinase inhibitors.

Rho kinase (ROCK) is a promising drug target for the treatment of many diseases including hypertension, multiple sclerosis, cancer, and glaucoma. The structure-activity relationships (SAR) around a series of tetrahydroisoquinolines were evaluated utilizing biochemical and cell-based assays to measure ROCK inhibition. These novel ROCK inhibitors possess high potency, high selectivity, and appropriate pharmacokinetic properties for glaucoma applications. The lead compound, 35, had subnanomolar potency in enzyme ROCK-II assays as well as excellent cell-based potency (IC(50) = 51 nM). In a kinase panel profiling, 35 had an off-target hit rate of only 1.6% against 442 kinases. Pharmacology studies showed that compound 35 was efficacious in reducing intraocular pressure (IOP) in rats with reasonably long duration of action. These results suggest that compound 35 may serve as a promising agent for further development in the treatment of glaucoma.