Electrophysiological properties of the L-type Ca(2+) current in cardiomyocytes from bluefin tuna and Pacific mackerel.

Tunas are capable of exceptionally high maximum metabolic rates; such capability requires rapid delivery of oxygen and metabolic substrate to the tissues. This requirement is met, in part, by exceptionally high maximum cardiac outputs, opening the possibility that myocardial Ca(2+) delivery is enhanced in myocytes from tuna compared with those from other fish. In this study, we investigated the electrophysiological properties of the cardiac L-type Ca(2+) channel current (I(Ca)) to test the hypothesis that Ca(2+) influx would be large and have faster kinetics in cardiomyocytes from Pacific bluefin tuna (Thunnus orientalis) than in those from its sister taxon, the Pacific mackerel (Scombe japonicus). In accordance with this hypothesis, I(Ca) in atrial myocytes from bluefin tuna had significantly greater peak current amplitudes and faster fast inactivation kinetics (-4.4 +/- 0.2 pA/pF and 25.9 +/- 1.6 ms, respectively) than those from mackerel (-2.7 +/- 0.5 pA/pF and 32.3 +/- 3.8 ms, respectively). Steady-state activation, inactivation, and recovery from inactivation were also faster in atrial myocytes from tuna than from mackerel. In ventricular myocytes, current amplitude and activation and inactivation rates were similar in both species but elevated compared with those of other teleosts. These results indicate enhanced I(Ca) in atrial myocytes from bluefin tuna compared with Pacific mackerel; this enhanced I(Ca) may be associated with elevated cardiac performance, because I(Ca) delivers the majority of Ca(2+) involved in excitation-contraction coupling in most fish hearts. Similarly, I(Ca) is enhanced in the ventricle of both species compared with other teleosts and may play a role in the robust cardiac performance of fishes of the family Scombridae.

Longitudinal variation in muscle protein expression and contraction kinetics of largemouth bass axial muscle.

The present study investigates muscle protein expression in largemouth bass Micropterus salmoides through intra- and intermyomeric comparisons of white muscle. Using denaturing SDS-polyacrylamide gel electrophoresis, muscle protein expression in the arm and cone regions of sequential myomeres was compared for three bass. Low percentage (4.75 %) polyacrylamide-SDS gels and cyanogen bromide (CNBr) peptide mapping revealed no obvious intramyomeric differences between the myosin heavy chains of the arm and cone regions. Electrophoresis of myofibrils and muscle homogenates on higher percentage gels also failed to demonstrate any significant differences between arm and cone regions in either the myosin light chains or any of the major insoluble and soluble contractile proteins. Two differences were discovered intermyomerically: (i) the ratio of two troponin T isoforms changed from head to tail and (ii) caudal muscle had a lower total parvalbumin content than rostral muscle. Since troponin T and parvalbumin have been implicated in the regulation of skeletal muscle kinetics, longitudinal variation in muscle contraction kinetics was predicted. Subsequent experiments revealed that bass rostral white muscle showed faster rates of activation and relaxation than more caudal muscle, as has been observed in white muscle of other fish species. Rostral-caudal variations in white muscle protein composition and contractile properties are predicted to affect patterns of power production during fast, unsteady swimming.

Altered expression of myosin heavy chain in the vastus lateralis muscle in patients with COPD.

This study was designed to further characterize peripheral skeletal muscle alterations in patients with chronic obstructive pulmonary disease (COPD) and to evaluate the possible relationship between myosin heavy chain (MyoHC) isoform expression and exercise tolerance in these individuals. MyoHC composition from biopsy of the vastus lateralis muscle was examined in 12 COPD patients (forced expiratory volume in one second (FEV1)=31+/-9% predicted, peak oxygen consumption (V'O2)=15+/-4 mL x kg(-1) x min(-1)) and 10 age-matched normal male subjects (peak V'O2=20+/-5 mL x kg(-1) x min(-1)). The proportion of MyoHC type I was smaller in COPD than in normals (27+/-17% versus 41+/-9%, p<0.05) with an increase in MyoHC type IIa (51+/-15% versus 39+/-9%, p<0.05) and the proportion of MyoHC type IIx being comparable between both groups. A significant relationship was found between peak V'Oo2 mL x kg(-1) x min(-1) and FEV1 % pred (r=0.91, p<0.0001) and the percentage of MyoHC type I (r=0.61, p=0.016). In stepwise multiple regression, only FEV1 % pred was found to be a significant determinant of peak V'O2 (p<0.0001). This variable explained 83% of the total variance of peak V'O2. In summary, this study showed considerable modifications in the phenotypic expression of the myosin heavy chain in the vastus lateralis muscle in patients with chronic obstructive pulmonary disease. An independent effect of myosin heavy chain expression on exercise capacity was not found. These results suggest that chronic inactivity and muscle deconditioning may not be the sole factors explaining peripheral muscle dysfunction in patients with chronic obstructive pulmonary disease.

Capillary density of skeletal muscle: a contributing mechanism for exercise intolerance in class II-III chronic heart failure independent of other peripheral alterations.

OBJECTIVES: The study was conducted to determine if the capillary density of skeletal muscle is a potential contributor to exercise intolerance in class II-III chronic heart failure (CHF). BACKGROUND: Previous studies suggest that abnormalities in skeletal muscle histology, contractile protein content and enzymology contribute to exercise intolerance in CHF. METHODS: The present study examined skeletal muscle biopsies from 22 male patients with CHF compared with 10 age-matched normal male control patients. Aerobic capacities, myosin heavy chain (MHC) isoforms, enzymes, and capillary density were measured. RESULTS: The patients with CHF demonstrated a reduced peak oxygen consumption when compared to controls (15.0+/-2.5 vs. 19.8+/-5.0 ml x kg(-1) x min(-1), p <0.05). Using cell-specific antibodies to directly assess vascular density, there was a reduction in capillary density in CHF measured as the number of endothelial cells/fiber (1.42+/-0.28 vs. 1.74+/-0.35, p = 0.02). In CHF, capillary density was inversely related to maximal oxygen consumption (r = 0.479, p = 0.02). The MHC IIx isoform was found to be higher in patients with CHF versus normal subjects (28.5+/-13.6 vs. 19.5+/-9.4, p <0.05). CONCLUSIONS: There was a significant reduction in microvascular density in patients with CHF compared with the control group, without major differences in other usual histologic and biochemical aerobic markers. The inverse relationship with peak oxygen consumption seen in the CHF group suggests that a reduction in microvascular density of skeletal muscle may precede other skeletal muscle alterations and play a critical role in the exercise intolerance characteristic of patients with CHF.

Rostral-caudal variation in troponin T and parvalbumin correlates with differences in relaxation rates of cod axial muscle.

Relaxation rate is an important determinant of axial muscle power production during the oscillatory contractions of undulatory locomotion. Recently, significant differences have been reported in the relaxation rates of rostral versus caudal white muscle fibers of the Atlantic cod Gadus morhua L. The present study investigates the biochemical correlates of this rostral-caudal physiological variation. Using denaturing gel electrophoresis, a series of fresh muscle samples from the dorsal epaxial muscle region was analyzed and several differences were detected. First, a gradual shift occurs in the expression of two troponin T isoforms along the length of the body. Second, rostral muscles were found to contain significantly greater amounts of parvalbumin than caudal muscles. Third, two soluble Ca(2+)-binding proteins, in addition to parvalbumin, were also detected in the rostral muscle samples yet were absent from the caudal samples. This suite of rostral-caudal variations provides a strong biochemical basis for regional differences in the relaxation rates of cod white muscle.

Acute hemolysis and renal failure with rifampicin-dependent antibodies after discontinuous administration.

Acute hemolysis as a reaction to rifampicin is extremely rare; case reports number less than 15. We recently evaluated a 65-year-old Cambodian refugee who self-regulated the use of rifampicin and isoniazid for pulmonary tuberculosis. Fifteen minutes after a single discontinuous oral dose, he developed flank pain, chills, rigors, vomiting, diarrhea, fever, and brown turbid urine. Laboratory tests at presentation showed acute intravascular hemolysis. Nonoliguric renal failure ensured, and he was transferred to our institution 2 days later. The patient was group A, Rh (D) positive, P1 negative with a cold autoantibody and cold anti-P1 alloantibody. The direct antiglobulin test was negative at the time of transfer. To evaluate the hemolysis, studies were done to test for rifampicin- or isoniazid-dependent antibodies. Rifampicin-dependent antibodies were detected in the antiglobulin phase with broad spectrum anti-human globulin, monospecific anti-gamma chain, and anti-complement antisera. Agglutination titers did not change after dithiothreitol reduction of the patient's serum. We conclude that this patient developed rifampicin-dependent IgG antibodies with complement-fixing capability. The presence of rifampicin-dependent antibodies should be suspected in a patient with hemolysis and/or renal failure taking rifampicin.