bluuebunny/crossref_metadata_2025 huggingface dataset:
Phosphorylation-activity relationships of AMPK and acetyl-CoA carboxylase in muscle
Authors: Park S. H. , Gammon S. R. , Knippers J. D. , Paulsen S. R. , Rubink D. S. , Winder W. W.
Published: March 2015

AMP-activated protein kinase (AMPK) is activated during muscle contraction in response to the increase in AMP and decrease in phosphocreatine (PCr). Once activated, AMPK has been proposed to phosphorylate a number of targets, resulting in increases in glucose transport, fatty acid oxidation, and gene transcription. Although it has been possible to directly observe phosphorylation of one of these targets, acetyl-CoA carboxylase (ACC) in vitro, it has been more difficult to obtain direct evidence of ACC phosphorylation in contracting skeletal muscle. In these experiments using a phosphoserine antibody to ACC and a phosphothreonine antibody to AMPK, evidence was obtained for phosphorylation and activation of ACC in vitro, in gastrocnemius muscle electrically stimulated at different frequencies, and in muscle from rats running on the treadmill. Significant negative linear correlations between phospho-ACC and ACC activity were observed in all models ( P < 0.01). The decline in ACC activity was related to the decrease in PCr and the rise in AMP. A relationship between phospho-AMPK (threonine 172) and activity of AMPK immunoprecipitated with anti-α2subunit antibody preparation was also observed. These data provide the first evidence of a direct link between extent of phosphorylation of these proteins at sites recognized by the antibodies and activity of the enzymes in electrically stimulated muscle and in muscle of rats running on the treadmill.
Selected Contribution: How does airway inflammation modulate asthmatic airway constriction? An antigen challenge study
Authors: Henderson A. C. , Ingenito E. P. , Atileh H. , Israel E. , Suki B. , Lutchen K. R.
Published: March 2015

During the late-phase (LP) response to inhaled allergen, mediators from neutrophils and eosinophils are released within the airways, resembling what occurs during an asthma attack. We compared the distribution of obstruction and degree of reversibility that follows a deep inspiration (DI) during early-phase (EP) and LP responses in nine asthmatic subjects challenged with allergen. Heterogeneity of constriction was assayed by determining frequency dependence of dynamic lung resistance and elastance, airway caliber by tracking airway resistance during a DI, and airway inflammation by measuring inflammatory cells in induced sputum postchallenge. Despite a paucity of eosinophils in the sputum at baseline (<1% of nonsquamous cells), asthmatic subjects showed a substantial EP response with highly heterogeneous constriction and reduced capacity to maximally dilate airways. The LP was associated with substantial airway inflammation in all subjects. However, five subjects showed only mild LP constriction, whereas four showed more marked LP constriction characterized by heterogeneous constriction similar to EP. Bronchoconstriction during LP was fully alleviated by administration of a bronchodilator. These findings, together with the impaired bronchodilatory response during a DI, indicate a physiological abnormality in asthma at the smooth muscle level and indicate that airway inflammation in asthma is associated with a highly nonuniform pattern of constriction. These data support the hypothesis that variability in responsiveness among asthmatic subjects derives from intrinsic differences in smooth muscle response to inflammation.
H2O2activates ryanodine receptor but has little effect on recovery of releasable Ca2+content after fatigue
Authors: Oba Toshiharu , Kurono Chieko , Nakajima Ritsuko , Takaishi Tetsuo , Ishida Kazuto , Fuller Geraldine A. , Klomkleaw Wuthichai , Yamaguchi Mamoru
Published: March 2015

We studied whether hydrogen peroxide (H2O2) at ≤10 μM activates the ryanodine receptor and decreases releasable Ca2+content in the sarcoplasmic reticulum after fatigue. Exposure of rabbit or frog skeletal muscle ryanodine receptors to 10 μM H2O2enhanced channel activity in lipid bilayers when the redox potential was defined at cis = −220 mV and trans = −180 mV. Channel activation by 10 μM H2O2was also observed when cispotential was set at −220 mV without defining transpotential, but the effect was less. Reduction of trans redox potential from −180 to −220 mV did not alter channel activity. H2O2at 500 μM failed to activate the channel when the redox potential was not controlled. Stimulation of the frog muscle fiber for 2 min (50 Hz, a duty cycle of 200 ms/s) decreased tetanus tension by ∼50%. After 1 min, tetanus recovered rapidly to ∼70% of control and thereafter slowly approached the control level. Amplitudes of caffeine- and 4-chloro- m-cresol-induced contractures were decreased after a 60-min rest. The decrease is not enhanced by exposure to 10 μM H2O2. These results suggest that H2O2markedly activates the ryanodine receptor under the redox control in vitro, but externally applied H2O2may not play an important role in the postfatigue recovery process.
Effect of puberty on coronary arteries from female pigs
Authors: Chatrath Ritu , Ronningen Karen L. , LaBreche Peter , Severson Sandra R. , Jayachandran Muthuvel , Bracamonte Margarita P. , Miller Virginia M.
Published: March 2015

Vascular function changes following loss of ovarian hormones in women at menopause and in experimental animals following surgical ovariectomy. Little is known about changes in vascular function during hormonal transition from sexual immaturity (juvenile) to sexual maturity. Therefore, experiments were designed to determine effects of natural puberty on vascular function in female pigs. Tissue was studied from eight juvenile (2-3 mo) and eight adult (5-6 mo) female pigs. Plasma nitric oxide (NO) was measured, and mRNA for endothelium-derived NO synthase (eNOS) and eNOS protein were determined in aortic endothelial cells. Rings of coronary arteries were suspended for measurement of isometric force in organ chambers. Serum 17β-estradiol levels were comparable in the two groups, whereas the arithmetic mean of progesterone levels was about two-thirds lower in adults compared with juvenile pigs. Plasma NO was significantly higher in juveniles compared with adults, but mRNA and protein for eNOS were comparable. In coronary arteries, an α2-adrenergic agonist caused greater endothelium-dependent relaxations in rings from juvenile compared with adult pigs. Relaxations to bradykinin were similar in arteries from both groups, but inhibition of NO reduced relaxations only in arteries from juvenile pigs. Relaxations from NO were greater in arteries from adult compared with juvenile female pigs. In conclusion, coronary arterial endothelial and smooth muscle responses are selectively modulated at puberty in female pigs. At maturity, plasma NO is reduced and sensitivity of the smooth muscle to exogenous NO is increased. Posttranscriptional regulation of eNOS protein may explain differences in NO bioavailability in juvenile pigs.
Lung and alveolar wall elastic and hysteretic behavior in rats: effects of in vivo elastase treatment
Authors: Brewer Kelly K. , Sakai Hiroaki , Alencar Adriano M. , Majumdar Arnab , Arold Stephen P. , Lutchen Kenneth R. , Ingenito Edward P. , Suki Béla
Published: March 2015

We investigated the relationship between the microscopic elastic and hysteretic behavior of the alveolar walls and the macroscopic mechanical properties of the whole lung in an in vivo elastase-treated rat model of emphysema. We measured the input impedance of isolated lungs at three levels of transpulmonary pressure (Ptp) and used a linear model to estimate the dynamic elastance and hysteresivity of the lungs. The elastance of the normal lungs increased steeply with Ptp, whereas this dependence diminished in the treated lungs. Hysteresivity decreased significantly with Ptp in the normal lungs, but this dependence disappeared in the treated lungs. To investigate the microscopic origins of these changes, the alveolar walls were immunofluorescently labeled in small tissue strips. By using a fluorescent microscope, the lengths and angular orientations of individual alveolar walls were followed during cyclic uniaxial stretching of the tissue strips. The microstrains (relative change in segment length) and changes in angle of the alveolar walls showed considerable heterogeneity, which was interpreted in terms of a network model. In the normal strips, the alveolar walls showed larger angular changes compared with the treated tissue, whereas the alveolar walls of the treated tissue tended to be more extensible. Hysteresis in the average angle change was also larger in the treated tissue than in the normal tissue. We conclude that the decreased Ptp dependence of elastance and the constant hysteresivity in the treated lungs are related to microstructural remodeling and network phenomena at the level of the alveolar walls.
Selected Contribution: Hypersensitivity of pulmonary C fibers induced by adenosine in anesthetized rats
Authors: Gu Qihai , Ruan Ting , Hong Ju-Lun , Burki Nausherwan , Lee Lu-Yuan
Published: March 2015

Compelling clinical evidence implicates the potential role of adenosine in development of airway hyperresponsiveness and suggests involvement of pulmonary sensory receptors. This study was carried out to determine the effect of a low dose of adenosine infusion on sensitivity of pulmonary C-fiber afferents in anesthetized open-chest rats. Infusion of adenosine (40 μg · kg-1· min-1iv for 90 s) mildly elevated baseline activity of pulmonary C fibers. However, during adenosine infusion, pulmonary C-fiber responses to chemical stimulants and lung inflation (30 cmH2O tracheal pressure) were markedly potentiated; e.g., the response to right atrial injection of capsaicin (0.25 or 0.5 μg/kg) was increased by more than fivefold (change in fiber activity = 2.64 ± 0.67 and 16.27 ± 3.11 impulses/s at control and during adenosine infusion, n = 13, P < 0.05), and this enhanced response returned to control in ∼10 min. The potentiating effect of adenosine infusion was completely blocked by pretreatment with 8-cyclopentyl-1,3-dipropylxanthine (100 μg/kg), a selective antagonist of the adenosine A1receptor, but was not affected by 3,7-dimethyl-1-propargylxanthine (1 mg/kg), an A2-receptor antagonist, or 3-ethyl-5-benzyl-2-methyl-4-phenylethynyl-6-phenyl-1,4-(±)-dihydropyridine-3,5-dicarboxylate (2 mg/kg), an A3-receptor antagonist. This potentiating effect was also mimicked by N6-cyclopentyladenosine (0.25 μg·kg-1·min-1for 90 s), a selective agonist of the adenosine A1receptor. In conclusion, our results showed that infusion of adenosine significantly elevated the sensitivity of pulmonary C-fiber afferents in rat lungs and that this potentiating effect is likely mediated through activation of the adenosine A1receptor.
Minor amounts of plasma medium-chain fatty acids and no improved time trial performance after consuming lipids
Authors: Vistisen Bodil , Nybo Lars , Xu Xuebing , Høy Carl-Erik , Kiens Bente
Published: March 2015

Medium-chain triacylglycerols (MCT) have a potential glycogen-saving effect during exercise due to rapid hydrolysis and oxidation. However, studies comparing intake of carbohydrates (CHO) plus 80–90 g MCT with intake of CHO alone have revealed different results. The present study tested performance after consumption of specific structured triacylglycerol, consisting of a mixture of medium-chain fatty acids and long-chain fatty acids, to prevent the adverse effects observed by MCT (pure medium-chain fatty acids) regarding gastrointestinal distress. Seven well-trained subjects cycled 3 h at 55% of maximum O2uptake during which they ingested CHO or CHO plus specific structured triacylglycerols. Immediately after the constant-load cycling, the subjects performed a time trial of ∼50-min duration. Breath and blood samples were obtained regularly during the experiment. Fatty acid composition of plasma triacylglycerols, fatty acids, and phospholipids was determined. Performance was similar after administration of CHO plus specific structured triacylglycerol [medium-, long-, and medium-chain fatty acid (MLM)] compared with CHO (50.0 ± 1.8 and 50.8 ± 3.6 min, respectively). No plasma 8:0 was detected in the plasma lipid classes, but the amount of phospholipid fatty acids was significantly higher after CHO+MLM compared with CHO intake. The lacking time trial improvement after intake of medium-chain fatty acids might be due to no available 8:0 in the systemic circulation. A higher level of plasma phospholipid fatty acids in the CHO+MLM compared with the CHO group was probably due to endogenous phospholipid release into chylomicrons.
Indomethacin impairs LPS-induced behavioral fever in toads
Authors: Bicego K. C. , Steiner A. A. , Antunes-Rodrigues J. , Branco L. G. S.
Published: March 2015

We tested the hypothesis that PGs mediate lipopolysaccharide (LPS)-induced behavioral fever in the toad Bufo paracnemis. Measurements of preferred body temperature (Tb) were performed with a thermal gradient. Toads were injected intraperitoneally with the cyclooxygenase inhibitor indomethacin (5 mg/kg), which inhibits PG biosynthesis, or its vehicle (Tris) followed 30 min later by LPS (0.2 and 2 mg/kg) into the lymph sac. LPS at the dose of 0.2 mg/kg caused a significant increase in Tb from 7 to 10 h after injection, and then Tb returned toward baseline values. LPS at the dose of 2 mg/kg produced a different pattern of response, with a longer latency to the onset of fever (10th h) and a longer duration (until the end of the experiment at the 15th h). Tris significantly attenuated the fever induced by LPS at 0.2 mg/kg, but not at 2 mg/kg. Moreover, indomethacin completely blocked the fever evoked by LPS (2 mg/kg). These results indicate that the behavioral fever induced by LPS in toads requires the activation of the COX pathway, suggesting that the involvement of PG in fever has an ancient phylogenetic history and that endogenous PGs raise the thermoregulatory set point to produce fever, because behavioral thermoregulation seems to be related to changes in the thermoregulatory set point.
Myotoxic effects of clenbuterol in the rat heart and soleus muscle
Authors: Burniston Jatin G. , Ng Yeelan , Clark William A. , Colyer John , Tan Lip-Bun , Goldspink David F.
Published: March 2015

Myocyte-specific necrosis in the heart and soleus muscle of adult male Wistar rats was investigated in response to a single subcutaneous injection of the anabolic β2-adrenergic receptor agonist clenbuterol. Necrosis was immunohistochemically detected by administration of a myosin antibody 1 h before the clenbuterol challenge and quantified by using image analysis. Clenbuterol-induced myocyte necrosis occurred against a background of zero damage in control muscles. In the heart, the clenbuterol-induced necrosis was not uniform, being more abundant in the left subendocardium and peaking 2.4 mm from the apex. After position (2.4 mm from the apex), dose (5 mg clenbuterol/kg), and sampling time (12 h) were optimized, maximum cardiomyocyte necrosis was found to be 1.0 ± 0.2%. In response to the same parameters (i.e., 5 mg of clenbuterol and sampled at 12 h), skeletal myocyte necrosis was 4.4 ± 0.8% in the soleus. These data show significant myocyte-specific necrosis in the heart and skeletal muscle of the rat. Such irreversible damage in the heart suggests that clenbuterol may be damaging to long-term health.
Muscle activity damps the soft tissue resonance that occurs in response to pulsed and continuous vibrations
Authors: Wakeling James M. , Nigg Benno M. , Rozitis Antra I.
Published: March 2015

This study tested the hypotheses that when the excitation frequency of mechanical stimuli to the foot was close to the natural frequency of the soft tissues of the lower extremity, the muscle activity increases 1) the natural frequency and 2) the damping to minimize resonance. Soft tissue vibrations were measured with triaxial accelerometers, and muscle activity was measured by using surface electromyography from the quadriceps, hamstrings, tibialis anterior, and triceps surae groups from 20 subjects. Subjects were presented vibrations while standing on a vibrating platform. Both continuous vibrations and pulsed bursts of vibrations were presented, across the frequency range of 10–65 Hz. Elevated muscle activity and increased damping of vibration power occurred when the frequency of the input was close to the natural frequency of each soft tissue. However, the natural frequency of the soft tissues did not change in a manner that correlated with the frequency of the input. It is suggested that soft tissue damping may be the mechanism by which resonance is minimized at heel strike during running.
Blood pressure response to chronic episodic hypoxia: the renin-angiotensin system
Authors: Fletcher Eugene C. , Orolinova Natalia , Bader Michael
Published: March 2015

By using an inspired oxygen fraction that produces oxyhemoglobin desaturation equivalent to that seen in human sleep apnea, we have demonstrated that 35 days of recurrent episodic hypoxia (every 30 s for 7 h/day) results in an 8–13 mmHg persistent increase in diurnal systemic mean arterial blood pressure (MAP) in rats. Blockade of angiotensin II receptors (AT1a) eliminates this response. Separate groups of male Sprague-Dawley rats were fed high-salt (8%), ad libitum-salt, or low-salt (0.1%) diets for 7 wk: 2 wk of wash-in for baseline blood pressure measurement and 5 wk of experimental conditions. Rats in each salt group were subjected to episodic hypoxia whereas controls remained unhandled under normoxic conditions. MAP remained at basal levels in all nonepisodic hypoxia controls as well as high-salt-diet episodic hypoxia-exposed rats. Ad lib and low-salt episodic hypoxia rats showed an increase in MAP from 106 and 104 mmHg at baseline to 112 and 113 mmHg, respectively ( P < 0.05). Whole kidney renin mRNA was suppressed in high-salt controls and episodic hypoxia rats, whereas kidney AT1a mRNA showed opposite changes. Suppression of the renin-angiotensin system with a high-salt diet blocks the increase in MAP in episodic hypoxia-challenged rats, in part by suppressing local tissue renin levels. Upregulation of the tissue angiotensin II system appears to be necessary for the chronic blood pressure changes that occur from episodic hypoxia.
Distal projection of insufflated gas during tracheal gas insufflation
Authors: Carter Christopher S. , Hotchkiss John R. , Adams Alexander B. , Stone Mary K. , Marini John J.
Published: March 2015

Tracheal gas insufflation (TGI) flushes expired gas from the ventilator circuitry and central airways, augmenting CO2 clearance. Whereas a significant portion of this washout effect may occur distal to the injection orifice, the penetration and mixing behavior of TGI gas has not been studied experimentally. We examined the behavior of 100% oxygen TGI injected at set flow rates of 1–20 l/min into a simulated trachea consisting of a smooth-walled, 14-mm-diameter tube. Models incorporating a separate coaxial TGI injector, a rough-walled trachea, and a bifurcated trachea were also studied. One-hundred percent nitrogen, representing expiratory flow, passed in the direction opposite to TGI at set flow rates of 1–25 l/min. Oxygen concentration within the “trachea” was mapped as a function of axial and radial position. Three consistent findings were observed: 1) mixing of expiratory and TGI gases occurred close to the TGI orifice; 2) the oxygenated domain extended several centimeters beyond the endotracheal tube, even at high-expiratory flows, but had a defined distal limit; and 3) more distally from the site of gas injection, the TGI gas tended to propagate along the tracheal wall, rather than as a central projection. We conclude that forward-directed TGI penetrates a substantial distance into the central airways, extending the compartment susceptible to CO2 washout.
Myofascial force transmission: muscle relative position and length determine agonist and synergist muscle force
Authors: Huijing Peter A. , Baan Guus C.
Published: March 2015

Equal proximal and distal lengthening of rat extensor digitorum longus (EDL) were studied. Tibialis anterior, extensor hallucis longus, and EDL were active maximally. The connective tissues around these muscle bellies were left intact. Proximal EDL forces differed from distal forces, indicating myofascial force transmission to structures other than the tendons. Higher EDL distal force was exerted (ratio ≈118%) after distal than after equal proximal lengthening. For proximal force, the reverse occurred (ratio ≈157%). Passive EDL force exerted at the lengthened end was 7–10 times the force exerted at the nonlengthened end. While kept at constant length, synergists (tibialis anterior + extensor hallucis longus: active muscle force difference ≈ −10%) significantly decreased in force by distal EDL lengthening, but not by proximal EDL lengthening. We conclude that force exerted at the tendon at the lengthened end of a muscle is higher because of the extra load imposed by myofascial force transmission on parts of the muscle belly. This is mediated by changes of the relative position of most parts of the lengthened muscle with respect to neighboring muscles and to compartment connective tissues. As a consequence, muscle relative position is a major codeterminant of muscle force for muscle with connectivity of its belly close to in vivo conditions.
Rate of NO scavenging alters effects of recombinant hemoglobin solutions on pulmonary vasoreactivity
Authors: Resta Thomas C. , Walker Benjimen R. , Eichinger Mark R. , Doyle Michael P.
Published: March 2015

Many hemoglobin-based oxygen carriers (HBOCs) produce systemic and pulmonary hypertension and may increase microvascular permeability as a consequence of nitric oxide (NO) scavenging. In this study, we examined the effects of two recombinant human hemoglobin solutions, rHb1.1 and rHb2.0 for injection (rHb2.0), with different rates of NO scavenging on vasoconstrictor reactivity and vascular permeability in isolated, saline-perfused rat lungs. We hypothesized that rHb1.1, a first-generation HBOC with an NO scavenging rate similar to that of native human hemoglobin, would exacerbate pulmonary vasoconstriction and permeability and that rHb2.0, a second-generation HBOC with an NO scavenging rate ∼20- to 30-fold lower than that of rHb1.1, would minimally influence these responses. Consistent with this hypothesis, rHb1.1 enhanced pulmonary vasoconstrictor reactivity to both hypoxia and thromboxane mimetic U-46619 in a dose-dependent fashion. In contrast, rHb2.0 produced little or no change in reactivity to these stimuli. Furthermore, whereas rHb1.1 abrogated pulmonary vasodilation to the NO-donor S-nitroso- N-acetyl-penicillamine (SNAP), dose-dependent responses to SNAP were preserved, albeit attenuated, in lungs treated with rHb2.0. Finally, the capillary filtration coefficient was unaltered by either rHb1.1 or rHb2.0. We conclude that pulmonary hemodynamic responses to rHb2.0 are greatly reduced compared with those observed with rHb1.1, consistent with rHb2.0 having a diminished capacity to scavenge NO. In addition, neither hemoglobin solution measurably altered microvascular permeability in this preparation.
Cluster analysis of muscle functional MRI data
Authors: Damon Bruce M. , Wigmore Danielle M. , Ding Zhaohua , Gore John C. , Kent-Braun Jane A.
Published: March 2015

Muscle functional magnetic resonance imaging (mfMRI) is frequently used to determine spatial patterns of muscle involvement in exercising humans. A frequent finding in mfMRI is that, even within synergistic muscle groups, signal intensity (SI) data from individual voxels can be quite heterogeneous. The purpose of this study was to develop a novel method for organizing heterogeneous mfMRI data into clusters whose members behave similarly to each other but distinctly from members of other clusters and apply it in studies of functional compartmentalization in the anterior compartment of the leg. An algorithm was developed that compared the SI time courses of adjacent voxels and grouped together voxels that were sufficiently similar. The algorithm's performance was verified by using simulated data sets with known regional differences in SI time courses that were then applied to experimental mfMRI data acquired from six male subjects (age 22.6 ± 0.9 yr, mean ± SE) who sustained isometric contractions of the dorsiflexors at 40% of maximum voluntary contraction. The experimental data were also characterized by using a traditional analysis (user-specified regions of interest from a single image), in which the relative change in SI and the contrast-to-noise ratio [CNR; 100%×(SIRESTING - SIACTIVE)/(noise standard deviation)] were measured. In general, clusters were found in areas in which the CNR exceeded 5. Cluster analysis made functional distinctions between regions of muscle that were not seen with traditional analysis. In conclusion, cluster analysis's use of the full SI time course provides more sensitivity to muscle functional compartmentation than traditional analysis.
Oxygen uptake kinetics for moderate exercise are speeded in older humans by prior heavy exercise
Authors: Scheuermann Barry W. , Bell Chris , Paterson Donald H. , Barstow Thomas J. , Kowalchuk John M.
Published: March 2015

This study examined the effect of heavy-intensity warm-up exercise on O2 uptake (V˙o 2) kinetics at the onset of moderate-intensity (80% ventilation threshold), constant-work rate exercise in eight older (65 ± 2 yr) and seven younger adults (26 ± 1 yr). Step increases in work rate from loadless cycling to moderate exercise (Mod1), heavy exercise, and moderate exercise (Mod2) were performed. Each exercise bout was 6 min in duration and separated by 6 min of loadless cycling.V˙o 2 kinetics were modeled from the onset of exercise by use of a two-component exponential model. Heart rate (HR) kinetics were modeled from the onset of exercise using a single exponential model. During Mod1, the time constant (τ) for the predominant rise in V˙o 2(τV˙o 2) was slower ( P < 0.05) in the older adults (50 ± 10 s) than in young adults (19 ± 5 s). The older adults demonstrated a speeding ( P < 0.05) of V˙o 2kinetics when moderate-intensity exercise (Mod2) was preceded by high-intensity warm-up exercise (τV˙o 2, 27 ± 3 s), whereas young adults showed no speeding of V˙o 2kinetics (τV˙o 2, 17 ± 3 s). In the older and younger adults, baseline HR preceding Mod2was elevated compared with Mod1, but the τ for HR kinetics was slowed ( P < 0.05) in Mod2only for the older adults. Prior heavy-intensity exercise in old, but not young, adults speeded V˙o 2 kinetics during Mod2. Despite slowed HR kinetics in Mod2in the older adults, an elevated baseline HR before the onset of Mod2 may have led to sufficient muscle perfusion and O2 delivery. These results suggest that, when muscle blood flow and O2 delivery are adequate, muscle O2consumption in both old and young adults is limited by intracellular processes within the exercising muscle.
Vascular response to infusions of a nonextravasating hemoglobin polymer
Authors: Matheson Barbara , Kwansa Herman E. , Bucci Enrico , Rebel Annette , Koehler Raymond C.
Published: March 2015

The clinical utility of cross-linked tetrameric hemoglobin solutions is limited by peripheral vasoconstriction thought to be due to scavenging of nitric oxide. In addition, transfusion of crude preparations of hemoglobin polymers can cause arterial hypertension. We tested the hypothesis that eliminating low-molecular-weight components from the polymer solution would prevent extravasation and its associated pressor response. A zero-link polymer of bovine hemoglobin was developed without chemical linkers left between the tetramers. Transfusion of unprocessed preparations of these polymers in rats resulted in appearance of the polymer in the renal hilar lymph. However, eliminating the low-molecular-weight components with a 300-kDa diafiltration resulted in an average hydrodynamic radius of 250 Å and in undetectable levels of polymer in hilar lymph. Exchange transfusion in anesthetized rats and cats and in awake cats produced no increase in arterial pressure. In anesthetized cats, exchange transfusion with an albumin solution reduced hematocrit from 30 to 18%, increased cerebral blood flow, and dilated pial arterioles. In contrast, reducing hematocrit by transfusing the diafiltered polymer did not increase cerebral blood flow as pial arterioles constricted. These results are consistent with the hypothesis that the increase in arterial pressure associated with cell-free hemoglobin transfusion depends on hemoglobin extravasation. Constriction observed in the cerebrovascular bed with a nonextravasating hemoglobin polymer at low hematocrit is presumably a regulatory response to prevent overoxygenation at low blood viscosity.
Attenuation of cardiovascular adaptations to exercise in frail octogenarians
Authors: Ehsani Ali A. , Spina Robert J. , Peterson Linda R. , Rinder Morton R. , Glover Kathryn L. , Villareal Dennis T. , Binder Ellen F. , Holloszy John O.
Published: March 2015

To determine the mechanisms underlying increased aerobic power in response to exercise training in octogenarians, we studied mildly frail elderly men and women randomly assigned to an exercise group ( n = 22) who participated in a training program of 6 mo of physical therapy, strength training, and walking followed by 3 mo of more intense endurance exercise at 78% of peak heart rate or a control sedentary group ( n = 24). Peak O2 consumption (V̇o2 peak) increased 14% in the exercise group ( P < 0.0001) but decreased slightly in controls. Training induced 14% increase ( P = 0.027) in peak exercise cardiac output (Q̇), determined via acetylene re-breathing, and no change in arteriovenous O2 content difference. The increase in Q̇ was mediated by increases in heart rate ( P = 0.009) and probably stroke volume ( P = 0.096). Left ventricular stroke work also increased significantly. In the men, the increase in V̇o2 peak was exclusively due to a large increase in peak Q̇ (22%). In the women, the gain in V̇o2 peak was due to small increases in Q̇ and O2 extraction from skeletal muscles. Pulse pressure normalized for stroke volume and arterial elastance during peak effort did not change with training. Controls showed no changes. The results suggest that, although frail octogenarians have a diminished capacity for improvement in aerobic power in response to exercise training, this adaptation is mediated mostly by an increase in Q̇ during peak effort. Furthermore, Q̇ likely plays a greater role in the adaptive increase in V̇o2 peak in old men than old women.
Strenuous but not moderate exercise increases the thrombotic tendency in healthy sedentary male volunteers
Authors: Cadroy Yves , Pillard Fabien , Sakariassen Kjell S. , Thalamas Claire , Boneu Bernard , Riviere Daniel
Published: March 2015

We have investigated the effect of moderate and strenuous exercise on experimental arterial thrombus formation in men. Thrombogenesis was measured in 15 sedentary healthy male volunteers at rest or immediately after two standardized exercise tests performed for 30 min on a bicycle ergometer. The exercises were performed at a constant load corresponding to either 50 or 70% maximal oxygen uptake. Thrombus formation was induced ex vivo by exposing a collagen-coated coverslip in a parallel plate perfusion chamber to native nonanticoagulated blood for 3 min. The shear rate at the collagen surface was 2,600 s−1. Platelet and fibrin deposition was quantified by immunoenzymatic methods. The results show that moderate exercise did not affect arterial thrombus formation. In contrast, platelet thrombus formation on collagen was increased on the average by 20% after 30 min at 70% maximal oxygen uptake ( P = 0.03). Fibrin deposition on collagen remained unchanged with exercise, regardless of its intensity. Thus, with the use of a clinically relevant human experimental model of thrombosis, the present study suggests that exercise of heavy intensity may increase the risk for arterial thrombogenesis in sedentary young healthy male volunteers.
Stimulation of regional lymphatic and blood flow by epicutaneous oxazolone
Authors: He Chufa , Young Alan J. , West Charles A. , Su Mei , Konerding Moritz A. , Mentzer Steven J.
Published: March 2015

The application of the epicutaneous antigen oxazolone results in persistent induration and erythema; however, the relative changes in lymph and blood flow in the inflammatory skin are largely unknown. To define the contribution of lymph and blood flow to the clinical appearance of cutaneous inflammation, we studied the sheep ear after the application of oxazolone. As a model for the study of these changes, the sheep ear had several experimental advantages: 1) a simplified superficial vascular network, 2) defined lymphatic drainage, and 3) an avascular and alymphatic cartilaginous barrier. Lymph flow was continuously monitored by cannulation of the prescapular efferent lymph duct. Blood flow, as reflected by cutaneous erythema, was noninvasively measured by use of a visible-spectrum spectrophotometer. The application of the epicutaneous oxazolone resulted in increased ear thickness for >7 days. The lymph flow from the oxazolone-stimulated ear peaked between 24 and 48 h after oxazolone stimulation. Spectrophotometric evaluation indicated that the cutaneous erythema peaked 72–96 h after application of oxazolone. Corrosion casting and scanning electron microscopy of the microcirculation at 96 h after antigen stimulation demonstrated significant dilatation of the superficial vascular network. These results suggest a biphasic response to oxazolone stimulation: 1) an early increase in vascular permeability associated with increased lymph flow and 2) a subsequent increase in relative blood flow associated with a dilated inflammatory microcirculation.
A mouse-based strategy for cyclophosphamide pharmacogenomic discovery
Authors: Watters James W. , Kloss Ellen F. , Link Daniel C. , Graubert Timothy A. , McLeod Howard L.
Published: March 2015

Genome-wide mapping approaches are needed to more fully understand the genetic basis of chemotherapy response. Because of technical and ethical limitations, cancer pharmacogenomics has not yet benefited from traditional robust familial genetic strategies. We have therefore explored the use of the inbred mouse as a genetic model system in which to study response to the cytotoxic agent cyclophosphamide. Multiple phenotypes have been assessed in response to cyclophosphamide in up to 19 inbred mouse strains, including in vitro hematopoietic progenitor cell toxicity and the mobilization of hematopoietic progenitor cells into peripheral blood. Hematopoietic progenitor cell toxicity in vitro varied 2-fold among strains, whereas in vivo progenitor cell mobilization varied almost 75-fold among strains. Males mobilized more hematopoietic progenitor cells than did females, and the low-mobilization phenotype was dominant to the high-mobilization phenotype in F1 hybrid animals. In an initial attempt to analyze candidate genes, genetic variation was assessed in three cytochrome P-450 genes involved in the metabolism of cyclophosphamide. Resequencing of eight strains identified 26 polymorphisms in these genes that may influence response to cyclophosphamide. Distinct regions of high- and low-polymorphism rates were identified, and two common haplotypes were shared among the strains for each gene that exhibited variation. This phenotypic and genotypic variation among inbred strains provides a framework for cyclophosphamide pharmacogenomic discovery.
Mechanical properties of the latissimus dorsi muscle after cyclic training
Authors: Askew Graham N. , Cox Valerie M. , Altringham John D. , Goldspink David F.
Published: March 2015

Cardiomyoplasty is a procedure developed to improve heart performance in patients suffering from congestive heart failure. The latissimus dorsi (LD) muscle is surgically wrapped around the failing ventricles and stimulated to contract in synchrony with the heart. The LD muscle is easily fatigued and as a result is unsuitable for cardiomyoplasty. For useful operation as a cardiac-assist device, the fatigue resistance of the LD muscle must be improved while retaining a high power output. The LD muscle of rabbits was subjected to a training regime in which cyclic work was performed. Training transformed the fiber-type composition from approximately equal proportions of fast oxidative glycolytic (FOG) and fast glycolytic (FG) fibers to one composed of almost entirely of FOG with no FG, which increased fatigue resistance while retaining rapid contraction kinetics. Muscle mass and cross-sectional area increased but power output decreased, relative to control muscles. This training regime represents a significant improvement in terms of preserving muscle mass and power compared with other training regimes, while enhancing fatigue resistance, although some fiber damage occurred. The power output of the trained LD muscle was calculated to be sufficient to deliver a significant level of assistance to a failing heart during cardiomyoplasty.
Interaction between fascicles and tendinous structures during counter movement jumping investigated in vivo
Authors: Kurokawa Sadao , Fukunaga Tetsuo , Nagano Akinori , Fukashiro Senshi
Published: March 2015

Behavior of fascicles and tendinous structures of the m. gastrocnemius medialis (MG) was quantitatively examined during human jumping in vivo. Eight male subjects performed maximal-effort counter movement jumping (CMJ) motions. Kinematic and kinetic data were obtained using a high-speed camera and a force platform. Behavior of fascicles and tendinous structures was determined using ultrasonography and electromyography. Although the muscle-tendon complex (MTC) shortened by only 1.6% during the downward phase of the counter movement, fascicles shortened as much as 10.4%. This shortening of fascicles caused elongation of tendinous structures by 2.2%. Although the MTC remained at almost constant length during the upward-I phase (–250 to –100 ms before toe-off), fascicles shortened by 19.2% of the initial length with an elongation of tendinous structures by 4.4%. The MTC shortened rapidly by 5.3% of the initial length during the upward-II phase (–100 to 0 ms), whereas fascicles shortened slightly during the first half of this phase and contracted in a quasi-isometric manner during the latter half of this phase. These findings implied that elastic energy was stored in tendinous structures throughout the latter half of the downward phase (1.0 J) and upward-I phase (5.6 J), which was thereafter rapidly released during the upward-II phase (3.8 J). It was found that muscle fibers of the MG were not stretched during counter movement; therefore, stretch reflex and potentiation of the contractile component of the MG might not contribute to the work enhancement in CMJ. It was suggested that the interaction between fascicles and tendinous structures was essential in a generation of higher joint power during the late push-off phase. This behavior of the MTC of the MG in CMJ was quite similar to what was observed in squat jumping performed without counter movement.
Invited Review: Role of insulin in translational control of protein synthesis in skeletal muscle by amino acids or exercise
Authors: Kimball Scot R. , Farrell Peter A. , Jefferson Leonard S.
Published: March 2015

Protein synthesis in skeletal muscle is modulated in response to a variety of stimuli. Two stimuli receiving a great deal of recent attention are increased amino acid availability and exercise. Both of these effectors stimulate protein synthesis in part through activation of translation initiation. However, the full response of translation initiation and protein synthesis to either effector is not observed in the absence of a minimal concentration of insulin. The combination of insulin and either increased amino acid availability or endurance exercise stimulates translation initiation and protein synthesis in part through activation of the ribosomal protein S6 protein kinase S6K1 as well as through enhanced association of eukaryotic initiation factor eIF4G with eIF4E, an event that promotes binding of mRNA to the ribosome. In contrast, insulin in combination with resistance exercise stimulates translation initiation and protein synthesis through enhanced activity of a guanine nucleotide exchange protein referred to as eIF2B. In both cases, the amount of insulin required for the effects is low, and a concentration of the hormone that approximates that observed in fasting animals is sufficient for maximal stimulation. This review summarizes the results of a number of recent studies that have helped to establish our present understanding of the interactions of insulin, amino acids, and exercise in the regulation of protein synthesis in skeletal muscle.
Rho inhibition decreases TNF-induced endothelial MAPK activation and monolayer permeability
Authors: Nwariaku Fiemu E. , Rothenbach Patricia , Liu Zijuan , Zhu Xudong , Turnage Richard H. , Terada Lance S.
Published: March 2015

Our laboratory previously demonstrated that MAPK activation is an important signal during cytokine-induced endothelial permeability (Nwariaku FE, Liu Z, Terada L, Duffy S, Sarosi G, and Turnage R. Shock 18: 82-85, 2002). Because GTP-binding proteins have been implicated in MAPK activation, we now hypothesize that the GTP-binding protein Rho is a mediator of TNF-induced MAPK activation and increased endothelial permeability. Transmonolayer permeability was assessed in human lung microvascular cells by measuring transmonolayer electrical resistance. MAPK activity was assessed by using a phospho-specific immunoprecipitation kinase assay and by comparing Western blots for phospho-MAPK with total MAPK. MAPK inhibitors used were SB-202190 and PD-098059, whereas Clostridium botulinum C3 transferase was used as a Rho inactivator. Rho-associated coiled-coil kinase was inhibited with Y-27632. TNF increased pulmonary endothelial permeability in vitro and caused a rapid, sustained increase in endothelial p38 and extracellular signal-regulated kinase MAPK activity. Inhibition of p38 and extracellular signal-regulated kinase MAPK with SB-202190 and PD-098059, respectively, decreased TNF-induced endothelial permeability. C3 transferase attenuated TNF-induced MAPK activation and blocked TNF-induced endothelial permeability. Finally, inhibition of Rho-associated coiled-coil kinase with Y-27632 prevented both MAPK activation and TNF-induced decreases in transmonolayer resistance. Rho acts upstream of mitogen-activated protein kinases in mediating TNF-induced pulmonary endothelial leak.
Sleep apnea and effect of chemostimulation on breathing instability in mice
Authors: Nakamura Akira , Fukuda Yasuichiro , Kuwaki Tomoyuki
Published: March 2015

Sleep apnea occurs in humans and experimental animals. We examined whether it also arises in adult mice. Ventilation in male adult 129/Sv mice was recorded concomitantly by electroencephalograms and electromyograms for 6 h by use of body plethysmography. Apnea was defined as cessation of plethysmographic signals for longer than two respiratory cycles. While mice breathed room air, 32.3 ± 6.9 (mean ± SE, n = 5) apneas were observed during sleep but not in quiet awake periods. Sleep apneas were further classified into two types. Postsigh apneas occurred exclusively during slow-wave sleep (SWS), whereas spontaneous apneas arose during both SWS and rapid eye movement sleep. Compared with room air (9.1 ± 1.4/h of SWS), postsigh apneas were more frequent in hypoxia (13.7 ± 2.1) and less frequent in hyperoxia (3.6 ± 1.7) and hypercapnia (2.8 ± 2.1). Our data indicated that significant sleep apnea occurs in normal adult mice and suggested that the mouse could be a promising experimental model with which to study the genetic and molecular basis of respiratory regulation during sleep.
Effect of contraction frequency on the contractile and noncontractile phases of muscle venous blood flow
Authors: Hogan Michael C. , Grassi Bruno , Samaja Michele , Stary Creed M. , Gladden L. B.
Published: March 2015

The purpose of this study was to test the hypothesis that increasing muscle contraction frequency, which alters the duty cycle and metabolic rate, would increase the contribution of the contractile phase to mean venous blood flow in isolated skeletal muscle during rhythmic contractions. Canine gastrocnemius muscle ( n = 5) was isolated, and 3-min stimulation periods of isometric, tetanic contractions were elicited sequentially at rates of 0.25, 0.33, and 0.5 contractions/s. The O2 uptake, tension-time integral, and mean venous blood flow increased significantly ( P < 0.05) with each contraction frequency. Venous blood flow during both the contractile (106 ± 6, 139 ± 8, and 145 ± 8 ml·100 g-1·min-1) and noncontractile phases (64 ± 3, 78 ± 4, and 91 ± 5 ml·100 g-1·min-1) increased with contraction frequency. Although developed force and duration of the contractile phase were never significantly different for a single contraction during the three contraction frequencies, the amount of blood expelled from the muscle during an individual contraction increased significantly with contraction frequency (0.24 ± 0.03, 0.32 ± 0.02, and 0.36 ± 0.03 ml·N-1·min-1, respectively). This increased blood expulsion per contraction, coupled with the decreased time in the noncontractile phase as contraction frequency increased, resulted in the contractile phase contribution to mean venous blood flow becoming significantly greater (21 ± 4, 30 ± 4, and 38 ± 6%) as contraction frequency increased. These results demonstrate that the percent contribution of the muscle contractile phase to mean venous blood flow becomes significantly greater as contraction frequency (and thereby duty cycle and metabolic rate) increases and that this is in part due to increased blood expulsion per contraction.
Efficient design of the diaphragm: distribution of blood flow relative to mechanical advantage
Authors: Johnson Robert L. , Hsia Connie C. W. , Takeda Shin-Ichi , Wait Juliette L. , Glenny Robb W.
Published: March 2015

The mammalian diaphragm is composed of two separate muscles (costal and crural) connected by a central tendon that serves as a piston head for drawing air into the lungs. The two muscles are described as having different embryological origins, segmental innervations, and mechanical functions [De Troyer A, Sampson M, Sigrist S, and Macklem PT. Science 213: 237–238, 1981; De Troyer A, Sampson M, Sigrist S, and Macklem PT. J Appl Physiol 53: 30–39, 1982]. On the basis of regional blood flow measurements, the two muscles appear to be nonuniformly recruited at rest, but we anticipated that the two muscles would become uniformly recruited at heavy exercise to efficiently support the high energy requirements of ventilation. We used fluorescent microspheres to measure regional blood flow within the two muscles as an index of muscle recruitment from rest to heavy treadmill exercise in well-trained foxhounds. However, the heterogeneity of blood flow at rest persisted as exercise workloads were increased. Blood flow per gram of muscle remained twofold greater in ventral than dorsal regions of both muscles from rest to heavy exercise. This pattern was matched by a twofold greater regional mechanical advantage in ventral than dorsal regions of the two muscles measured anatomically. Hence blood flow was preferentially and efficiently distributed to those regions capable of generating the greatest inspiratory power independent of muscle mass. The two muscles were recruited from rest to heavy exercise as a single functional unit, not as two muscles under separate control.