Timo Takala

Role of blood flow in regulating insulin-stimulated glucose uptake in humans. Studies using bradykinin, [15O]water, and [18F]fluoro-deoxy-glucose and positron emission tomography.

Defects in insulin stimulation of blood flow have been used suggested to contribute to insulin resistance. To directly test whether glucose uptake can be altered by changing blood flow, we infused bradykinin (27 microgram over 100 min), an endothelium-dependent vasodilator, into the femoral artery of 12 normal subjects (age 25+/-1 yr, body mass index 22+/-1 kg/m2) after an overnight fast (n = 5) and during normoglycemic hyperinsulinemic (n = 7) conditions (serum insulin 465+/-11 pmol/liter, 0-100 min). Blood flow was measured simultaneously in both femoral regions using [15O]-labeled water ([15O]H2O) and positron emission tomography (PET), before and during (50 min) the bradykinin infusion. Glucose uptake was measured immediately after the blood flow measurement simultaneously in both femoral regions using [18F]-fluoro-deoxy-glucose ([18F]FDG) and PET. During hyperinsulinemia, muscle blood flow was 58% higher in the bradykinin-infused (38+/-9 ml/kg muscle x min) than in the control leg (24+/-5, P<0.01). Femoral muscle glucose uptake was identical in both legs (60.6+/-9.5 vs. 58.7+/-9.0 micromol/kg x min, bradykinin-infused vs control leg, NS). Glucose extraction by skeletal muscle was 44% higher in the control (2.6+/-0.2 mmol/liter) than the bradykinin-infused leg (1.8+/-0.2 mmol/liter, P<0.01). When bradykinin was infused in the basal state, flow was 98% higher in the bradykinin-infused (58+/-12 ml/kg muscle x min) than the control leg (28+/-6 ml/kg muscle x min, P<0.01) but rates of muscle glucose uptake were identical in both legs (10.1+/-0.9 vs. 10.6+/-0.8 micromol/kg x min). We conclude that bradykinin increases skeletal muscle blood flow but not muscle glucose uptake in vivo. These data provide direct evidence against the hypothesis that blood flow is an independent regulator of insulin-stimulated glucose uptake in humans.

Effects of power training on muscle structure and neuromuscular performance

The present study examines changes in muscle structure and neuromuscular performance induced by 15 weeks of power training with explosive muscle actions. Twenty‐three subjects, including 10 controls, volunteered for the study. Muscle biopsies were obtained from the gastrocnemius muscle before and after the training period, while maximal voluntary isometric contractions (MVC) and drop jump tests were performed once every fifth week. No statistically significant improvements in MVC of the knee extensor (KE) and plantarflexor muscles were observed during the training period. However, the maximal rate of force development (RFD) of KE increased from 18 836±4282 to 25 443±8897 N (P<0.05) during the first 10 weeks of training. In addition, vertical jump height (vertical rise of the center of body mass) in the drop jump test increased significantly (P<0.01). Simultaneously, explosive force production of KE muscles measured as knee moment and power increased significantly; however, there was no significant change (P>0.05) in muscle activity (electromyography) of KE. The mean percentage for myosin heavy chain and titin isoforms, muscle fiber‐type distributions and areas were unchanged. The enhanced performance in jumping as a result of power training can be explained, in part, by some modification in the joint control strategy and/or increased RFD capabilities of the KE.

Reduced stretch-reflex sensitivity after exhausting stretch-shortening cycle exercise

The stretch-shortening cycle (SSC) is an effective and natural form of muscle function but, when repeated with sufficient intensity or duration, it may lead to muscle damage and functional defects. A reduced tolerance to impact has been reported, which may be partly attributed to a reduced stretch-reflex potentiation. The aim of the present study was to examine the influence of SSC-induced metabolic fatigue and muscle damage on the efficacy of stretch reflexes, as judged by the electromyograph (EMG) response of two shank muscles (lateral gastrocnemius LG, soleus SOL) to controlled ramp stretches. These EMG responses were recorded before and immediately after exhausting SSC-type leg exercise and 2 h, 2 days and 4 days later. Serum concentrations of creatine kinase ([CK]), myoglobin and lactate were measured repetitively along the protocol. Two maximal vertical drop jumps and counter-movement jumps were performed after each reflex test. The exhausting SSC-type exercise induced an immediate reduction (P < 0.05) with a delayed short-term recovery of the LG peak-to-peak reflex amplitude. This was not accompanied by significant changes in the reflex latency. The drop jump performance remained slightly but significantly reduced (P < 0.05) until the 2nd day postexercise. Peak [CK] appeared for all the subjects on the 2nd day, suggesting the presence of muscle damage. The increase in [CK] between the 2nd h and the 2nd day postexercise was found to be negatively related (P < 0.001) to the relative changes in the drop jump height. Furthermore, a significant relationship (P < 0.05) was found between recovery of the stretch reflex in LG and the decrease of [CK] between the 2nd and the 4th day. hese findings support the hypothesis of a reduced stretch-reflex sensitivity. While the exact mechanisms of the reflex inhibition remain unclear, it is emphasized that the delayed recovery of the reflex sensitivity could have resulted from the progressive inflammation that develops in cases of muscle damage.

THE DISRUPTION OF MYOFIBRE STRUCTURES IN RAT SKELETAL MUSCLE AFTER FORCED LENGTHENING CONTRACTIONS

Abstract Specific antibodies against structural proteins (actin, desmin, dystrophin, fibronectin) of muscle fibres were used to study the effect of forced lengthening contractions on muscle microarchitecture. Tibialis anterior (TA) muscle of male Wistar rats were subjected to 240 forced lengthening contractions. At consecutive time points (0, and 6 h, 2, 4, and 7 days) after stimulation, the TA muscle was excised for biochemical and histological assays. β-Glucuronidase activity, a quantitative indicator of muscle damage, showed increased values 2–7 days after the lengthening, peaking on day 4 (11.7-fold increase). A typical course of histopathological changes (myofibre swelling, necrosis and regeneration) was observed. In immunohistochemistry, the earliest abnormality observed was discontinuous dystrophin staining in some swollen fibres immediately after commencement of exercise, while at the same time no alterations occurred in the staining of the other antibodies studied. Six hours later, all the swollen fibres were uniformly desmin as well as dystrophin negative. The great majority, but not all, of the swollen fibres showed disorganized actin staining and intramyocellular localization of fibronectin. The early phase disruption of myofibre structures as measured in this study provides evidence of their central role following damage in skeletal muscle. These results suggest that the sequence of structural changes in the route to muscle fibre necrosis in injury induced by forced lengthening contraction originates in the disruption of the plasma membrane and the intermediate filament, which leads to disturbances in the myofibrillar system.

Warm underwater water-jet massage improves recovery from intense physical exercise

The effects of warm underwater water-jet massage on neuromuscular functioning, selected biochemical parameters (serum creatine kinase, lactic dehydrogenase, serum carbonic anhydrase, myoglobin, urine urea and creatinine) and muscle soreness were studied among 14 junior track and field athletes. Each subject spent, in a randomized order, two identical training weeks engaged in five strength/power training sessions lasting 3 days. The training weeks differed from each other only in respect of underwater water jet massage treatments. These were used three times (20 min each) during the treatment week and not used during the control week. During the treatment week continuous jumping power decreased and ground contact time increased significantly less (P < 0.05) and serum myoglobin increased more than during the control week. It is suggested that underwater water-jet massage in connection with intense strength/power training increases the release of proteins from muscle tissue into the blood and enhances the maintenance of neuro-muscular performance capacity.

Regulation of ventricular atrial natriuretic peptide release in hypertrophied rat myocardium. Effects of exercise.

Left ventricular hypertrophy is characterized by stimulation of ventricular synthesis of atrial natriuretic peptide (ANP). This study was designed to test the hypothesis that the increased ventricular ANP levels participate in the release of ANP into the circulation. Swimming was used as a physiologic model to induce ANP release from the heart, and atrial and ventricular levels of immunoreactive ANP (IR-ANP) and ANP messenger RNA (mRNA) were measured simultaneously in the spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats at rest and after swimming. IR-ANP concentration in the left ventricle of 1-year-old SHR with severe left ventricular hypertrophy was increased in association with the augmentation of ANP mRNA levels, whereas right ventricular levels of ANP were reduced in SHR compared with normotensive controls. A 30-minute exercise in hypertensive and in normotensive rats resulted in marked increases in mean arterial pressure, heart rate, plasma catecholamine levels, blood lactate levels, and plasma IR-ANP concentration. The increased ANP secretion was associated with a decrease in left (34-39%) and right (24%) ventricular concentration of IR-ANP; transmurally, this depletion of ventricular IR-ANP was greatest (28%) in the endocardial layer of the left ventricle of SHR. No significant differences were noted in total atrial and left or right auricular IR-ANP concentration between SHR and WKY rats or between the resting and swimming rats. When studied in vitro with an isolated, perfused heart preparation, the hypertrophic ventricular tissue after atrialectomy secreted more ANP into the perfusate than did control hearts; in SHR, ventricles contributed 28% of the total ANP release to perfusate, and in normotensive control rats, ventricles contributed 8%. These studies show that stimulated release of ANP is associated with depletion of endocardial left ventricular stores. The amount of ANP released in vitro and in vivo correlated with the degree of hypertrophy of the ventricle. Finally, the phorbol ester, known to increase ANP secretion from intact perfused hearts, had only a limited effect on ANP release after atrialectomy, suggesting that the secretion of ANP from ventricular cells may be mainly of the constitutive type.

Increased mRNAs for procollagens and key regulating enzymes in rat skeletal muscle following downhill running.

Abstract The purpose of the study was to investigate pre-translational regulation of collagen expression after a single bout of exercise. We analysed steady-state messenger ribonucleic acid (mRNA) levels for collagen types I, III and IV, α- and β-subunits of prolyl 4-hydroxylase and lysyl oxidase (enzymes modifying procollagen chains), and enzyme activity of prolyl 4-hydroxylase from rat soleus muscle (MS) and the red parts of quadriceps femoris muscle (MQF) after 12 h and after 1, 2, 4, 7 and 14 days of downhill (–13.5°) treadmill running at a speed of 17 m·min–1 for 130 min. Histological and biochemical assays revealed exercise-induced muscle damage in MQF but not MS. Steady-state mRNA levels for the α- and β-subunits of prolyl 4-hydroxylase in MQF, lysyl oxidase in MS and MQF were increased 12 h after running, whereas prolyl 4-hydroxylase activity did not increase until 2 days after exercise. The mRNA levels for the fibrillar collagens (I and III) and basement membrane type IV collagen significantly increased 1 day and 12 h after exertion, respectively. Peak mRNA levels were observed 2–4 days after running, the increases being more pronounced in MQF than in MS. No significant changes were observed in types I or III collagen at the protein level. Strenuous downhill running thus causes an increase in gene expression for collagen types I and III and their post-translational modifying enzymes in skeletal muscle in a co-ordinated manner. These changes, together with the increased gene expression of type IV collagen, may represent the regenerative response of muscle extracellular matrix to exercise-induced injury and an adaptive response to running exertion.

Biochemical composition of muscle extracellular matrix : the effect of loading

Collagen plays an important role in skeletal muscle both during muscle differentiation and normal muscle growth, and also serves a role as a supportive structure. It is the most abundant protein of the extracellular matrix and of the 19 distinct collagen types, types I, III, IV and V are the dominating ones in skeletal muscle. Both collagen synthesis as well as degradation is influenced by either physical loading or immobilization in skeletal muscle, and recent methods have allowed for greater understanding of the posttranslational processing of collagen.

Collagen synthesis and types I, III, IV, and VI collagens in an animal model of disc degeneration.

Study Design. The present study sought to elucidate the changes that occur in collagen chemistry in the early phases of disc degeneration. Objective. To monitor the healing process of the injured anulus fibrosus and the secondary degenerative reactions in the nucleus pulposus. Summary of Background Data. Despite the importance of collagen chemistry under pathologic conditions in the intervertebral disc, knowledge of this aspect is very limited. Methods. Fourteen pigs were stabbed with a scalpel blad in the anterior part of the anulus fibrosus of a lumbar disc. The animals were killed 2 weeks to 5 months after injury. The activities of prolyl 4-hydroxylase adn galactosylhydroxylysyl glucosyltransferase, the total collagen content, adn staining patterns for Types I, III, IV, and VI collagens were analyzed from different parts of the disc. Results. The most active phase of the healing process, assessed from the activities of enzymes involved in collagen biosynthesis, took place during the first month postoperatively. The anular lesion was found to cicatrize through formation of disorganized granulation tissue in which Type I, III, and, IV collagens were depostited. In the nucleus pulposus, activities of pralyl 4-hydroxylase and galactosylhydroxylysyl glucosyltransferase and total collagen content increased, and the originally rounded cells became more elongated, resembling fibroblasts. Conclusions. The results of this study suggest that the altered composition of collagens observed in the degenerate porcine nucleus pulposus results from changes in cell phenotype: Notochondral cells were replaced by fibroblast-like cells. It is likely that trauma to the anulus fibrosus can initiate a progressive degenerative process in the disc tissue.

Protein synthesis in the isolated perfused rat heart. Effects of mechanical work load, diastolic ventricular pressure and coronary pressure on amino acid incorporation and its transmural distribution into left ventricular protein.

SummaryThe mechanisms involved in hypertrophy of the left ventricle were studied in Langendorff-perfused rat hearts by measuring the ventricular protein synthesis and its transmural distribution and by differentiating between the effects of changes in mechanical work load, intraventricular and coronary pressures. An increase in the aortic pressure from 7.85 kPa (80 cm of water) to 19.6 kPa (200 cm of water) in beating hearts increased phenylalanine incorporation into left ventricular protein from 1.4 to 2.0 μmol/g protein (p<0.02) during a two-hour perfusion. The protein synthesis was transmurally evenly distributed. A similar elevation in the perfusion pressure in potassium-arrested hearts caused an increase in phenylalanine incorporation from 1.5 to 1.9 μmol/ (p<0.05) when the intraventricular pressure was adjusted to zero, indicating that the increase in aortic (coronary) pressure and not the work loadper se was the reason for increased protein synthesis. Elevation of the end-diastolic pressure from zero to ∼ 2 kPa in beating hearts at an aortic pressure of 7.85 kPa, or from 7.85 kPa to 17.3 kPa in K+-arrested hearts, at an aortic pressure of 19.6 kPa caused a significant reduction in subendocardial protein synthesis, whereas subepicardial phenylalanine incorporation was at most only slightly affected. The energetic parameters, oxygen consumption, output of vasoactive purine compounds and distribution of coronary flow indicate that the increase in protein synthesis via the elevation in aortic pressure was not due to the abolition of partial anoxia, whereas the same parameters indicate that the transmural gradient in protein synthesis observed under certain conditions was due to subendocardial ischemia when the intraventricular pressure approached the aortic pressure in arrested hearts, which are evidently of restricted use for extended periods without special measures to limit the build-up of intraventricular pressure.ZusammenfassungDie Mechanismen der Hypertrophie der linken Herzkammer und die Unterschiede zwischen der Wirkung der mechanischen Arbeit, des intraventrikulären und koronararteriellen Druckes wurden in Langendorff-perfundierten Rattenherzen untersucht. Bei einer Erhöhung des Perfusionsdruckes von 7,85 kPA (80 cm Wasser) auf 19,6 kPa (200 cm Wasser) bei schlagenden Herzen vermehrte sich die Inkorporation der Aminosäure Phenylalanin von 1,4 auf 2,0 μmol/g Protein (p<0,02) während einer zweistündigen Perfusion. Die Proteinsynthese zeigte eine gleichmäßige transmurale Verteilung. Eine gleiche Erhöhung des Perfusionsdruckes in mit Kalium stillgestellten Herzen vermehrte die Inkorporation von Phenylalanin von 1,5 auf 1,9 μmol/g (p<0,05) wenn der intraventrikuläre Druck auf Null festgestellt wurde, woraus hervorgeht, daß die Erhöhung des Perfusionsdruckes und nicht die mechanische Arbeit per se die Ursache der vergrößerten Proteinsynthese war. Erhöhung des enddiastolischen Druckes schlagender Herzen von Null auf 2,1 kPa bei einem Perfusionsdruck von 7,85 kPa oder Steigerung des Kammerdrucks K-arretierter Herzen von 7,85 kPa auf 17,3 kPa bei einem Perfusionsdruck von 19,6 kPa verursachte eine wesentliche Verminderung der subendokardialen Proteinsynthese, während sich die subepikardiale Phenylalanininkorporation nur wenig veränderte.Die Messung der energetischen Parameter, des Sauerstoffverbrauches und der Produktion der vasoaktiven Purinverbindungen sowie der transmuralen Verteilung der Myokarddurchblutung zeigten, daß die von dem erhöhten Perfusionsdruck verursachte Steigerung der Proteinsynthese nicht durch Beseitigung eines partiellen Sauerstoffmangels resultierte. Die gleichen Parameter sprechen andererseits dafür, daß der transmurale Gradient der Proteinsynthese unter bestimmten Bedingungen durch eine subendokardiale Ischämie bedingt war, wenn sich nämlich beim stillstehenden Herzen der Ventrikelinnendruck dem Aortendruck (Perfusionsdruck) nähert; stillstehende Herzen dürfen daher nicht ohne spezielle Regulierung des intraventrikulären Druckes über längere Zeit perfundiert werden.