Epithelia of the ovine and bovine forestomach express basolateral maxi-anion channels permeable to the anions of short-chain fatty acids

It has long been established that the absorption of short-chain fatty acids (SCFA) across epithelia stimulates sodium proton exchange. The apically released protons are not available as countercations for the basolateral efflux of SCFA anions and a suitable transport model is lacking. Patch clamp and microelectrode techniques were used to characterize an anion conductance expressed by cultured cells of the sheep and bovine rumen and the sheep omasum and to localize the conductance in the intact tissue. Cells were filled with a Na-gluconate solution and superfused with sodium salts of acetate, propionate, butyrate, or lactate. Reversal potential rose and whole cell current at +100 mV decreased with the size of the anion. Anion-induced currents could be blocked by diisothiocyanato-stilbene-2,2′-disulfonic acid (DIDS), NPPB (200?μmol l^?1), or pCMB (1 mmol l^?1). In patches of bovine ruminal cells, single channels were observed with a conductance for chloride (327?±?11 pS), acetate (115?±?8 pS), propionate (102?±?10 pS), butyrate (81?±?2 pS), and gluconate (44?±?3 pS). Channels expressed by sheep rumen and omasum were similar. Microelectrode experiments suggest basolateral localization. In conclusion, forestomach epithelia express basolateral maxi-anion channels with a permeability sequence of chloride?>?acetate?>?propionate?>?butyrate. SCFA absorption may resemble functionally coupled transport of NaCl, with the Na⁺/K⁺-ATPase driving the basolateral efflux of the anion through a channel. Since protons are apically extruded, the model accurately predicts that influx of buffers with saliva is essential for the pH homeostasis of the ruminant forestomach.     

Pannexin 1 forms an anion-selective channel

Pannexin 1 (Panx1) is expressed in various mammalian tissues including the brain and immune cells. Here, we present evidence that Panx1 when expressed in mammalian cells, forms anion-selective channels, with a rank order of permeabilities: NO₃⁻ > I⁻ > Br⁻ > Cl⁻ > F⁻ ≫ aspartate⁻ ≈ glutamate⁻ ≈ gluconate⁻. Single-channel Panx1-mediated currents have a unitary conductance around 68 pS. Our results show that Panx1 assembles into a membrane anion channel with a relatively low single-channel conductance.     

Haematological and serum biochemical profile of the upside-down catfish, Synodontis membranacea Geoffroy Saint Hilaire from Jebba Lake, Nigeria

Haematological and serum biochemical studies of natural population of Synodontis membranacea from Jebba Lake, North Central Nigeria were investigated in order to establish their mean and reference values. Bi-monthly collection of 1,408 live fish samples was carried out between April 2002 and March 2004, using gill nets of various mesh sizes ranging from 5.08 to 10.16 cm. The mean baseline value established for species-specific haematological and serum biochemical parameters were red blood cell (RBC) 3.83 ± 1.49 × 10¹² l⁻¹, haemoglobin (HB) 8.38 ± 1.96 g dl⁻¹, and packed cell volume (PCV) 25.65 ± 5.89%; mean cell volume 78.25 ± 37.90 fl; mean cell haemoglobin (MCH) 33.04 ± 12.50 pg; mean cell haemoglobin concentration 26.53 ± 15.18 g dl⁻¹; white blood cell (WBC) 315.65 ± 95.37 × 10⁻⁹; agranulocytes (Agr) 82.07 ± 11.38%; monocytes (Mon) 6.37 ± 3.01%; lymphocytes (Lym) 76.49 ± 10.81%; granulocytes (Gran) 40.28 ± 17.48%; neutrophils (Neut) 24.42 ± 10.68%; eosinophils (Eos) 16.14 ± 8.25%; basophils 0.09 ± 0.04%; protein 40.19 ± 7.45 g l⁻¹; albumin 19.78 ± 5.67 g l⁻¹; creatinine 49.71 ± 16.15 μmol l⁻¹; urea 3.05 ± 0.67 nmol l⁻¹; uric acid 0.76 ± 0.33 nmol l⁻¹; glucose 4.24 ± 1.74 mmol l⁻¹; cholesterol 8.46 ± 2.27 mmol l⁻¹; calcium 2.35 ± 0.94 mmol l⁻¹; potassium 13.36 ± 4.45 mmol l⁻¹; sodium 139.39 ± 23.19 mmol l⁻¹; alanine aminotransferase (ALT) 11.79 ± 2.67 U l⁻¹; aspartate aminotransferase 16.80 ± 4.73 U l⁻¹; and alkaline phosphatase 63.01 ± 20.44 U l⁻¹. Only three of these parameters (i.e. neutrophil, glucose and potassium) differed significantly (P > 0.05) on gender basis. Pearson’s correlation coefficients indicated significant relationship of standard length and total weight with RBC, PCV, HB, WBC, Agr, Mon, Lym, Gran, Neut, Eos, sodium, and ALT only. The study has provided baseline haematological and biochemical data for use in health monitoring and productivity of S. membranacea, which would be of great value for future comparative surveys in this era of increased fish culture in Nigeria.     

The effects of 3000-m swimming on subsequent 3-h cycling performance: implications for ultraendurance triathletes

The purpose of this study was to examine the physiological effects of 3000-m swimming on subsequent 3-h cycling time trial performance in ultraendurance triathletes. Eight highly trained ultraendurance triathletes [mean (SEM) age 34 (2) years, body fat 12.5 (0.8)%, maximum oxygen consumption 63.2 (2.1) ml · kg⁻¹ · min⁻¹] completed two randomly assigned trials 1 week apart. The swim/bike trial (SB) involved 3000 m of swimming [min:s 52:28 (1:48)] immediately followed by a 3-h cycling performance at a self-selected time-trial pace. The control trial (CON) consisted of an identical 3-h cycling time trial but without prior swimming. Subjects consumed an 8% carbohydrate (CHO)/electrolyte beverage during both trials at the rate of 60 g CHO · h⁻¹ and 1 l · h⁻¹. No significant differences were evident between CON and SB on the dependent measures (CON vs SB): power output [W˙, 222 (14) W vs 212 (13) W], heart rate [f _c, 147 (5) beats · min⁻¹ vs 143 (4) beats · min⁻¹; %f _cmax 80.0 (1.6)% vs 78.4 (1.5)%], oxygen uptake [3.10 (0.12) l · min⁻¹ vs 2.97 (0.15) l · min⁻¹], minute ventilation [82.5 (4.4) l · min⁻¹ vs 77.3 (3.7) l · min⁻¹], rating of perceived exertion [14.6 (0.4) vs 14.0 (0.1)], blood lactate [6.1 (0.5) mmol · l⁻¹ vs 4.8 (0.5) mmol · l⁻¹], and blood glucose [5.0 (0.2) mmol · l⁻¹ vs 5.3 (0.1) mmol · l⁻¹; all non-significant at the P > 0.05 level]. However, the CON respiratory exchange ratio was significantly greater than for SB [0.91 (0.01) vs 0.89 (0.01); P < 0.05], suggesting that the SB trial required a greater reliance on lipid as a fuel substrate. Hence, the main finding in the present study was that 3000 m of swimming had no significant performance effect (in terms of W˙) on subsequent 3-h cycling performance in ultraendurance triathletes. Accepted: 2 March 2000     

Chloride channels in the luminal membrane of rat pancreatic acini

 Pancreatic acini secrete Na⁺, Cl^–and H₂O in response to secretagogues such as acetylcholine. Cl^–channels in the luminal membrane are a prerequisite for this secretion. The properties of the corresponding conductance have previously been examined using whole-cell recordings. The present study attempts to examine the properties of the single channels in cell-attached and cell-free excised patches from the luminal membrane. To this end the pipettes were filled with an N-methyl-D-glucamine (NMDG⁺) chloride/gluconate solution. The voltage-clamp range was chosen to be pipette positive (cell negative, –60 to –130 mV) in order to increase the driving force for outward Cl^–currents. Under resting conditions cell attached luminal patches had very few single-channel currents (12 out of 45 experiments). Their incidence was sharply increased by carbachol (CCH, 1 μmol/l) in 41 out of 45 experiments. The single-channel conductance of these channels was 1.97 ± 0.05 pS. The properties of these channels in excised patches were examined further: their single-channel conductance was 2.2 ± 0.07 pS (n = 59) and their conductance selectivity was I^– > Br^– > Cl^– >> gluconate. None of the typical Cl^–channel blockers (DIDS, NPPB, glibenclamide 100 μmol/l) blocked these channels. It is concluded that the luminal membrane of the rat pancreatic acinus possesses Cl^–channels with very low conductance which are activated by carbachol. Received: 31 January 1997 / Received after revision: 26 February 1997 / Accepted: 5 March 1997     

In vivo 4-androstene-3,17-dione and 4-androstene-3β,17β-diol supplementation in young men

To determine if known androgenic hormone precursors for testosterone in the androgen pathway would be readily transformed to testosterone, eight male subjects [mean age 23.8 (SEM 3) years, bodymass 83.1 (SEM 8.7) kg, height 175.6 (SEM 8.5) cm] underwent a randomized, double-blind, cross-over, placebo-controlled oral treatment with 200 mg of 4-androstene-3,17-dione (Δ4), 4-androstene-3β,17β-diol (Δ4Diol), and placebo (PL). The periods of study were separated by 7 days of washout. Blood was drawn at baseline and subsequently every 30 min for 90 min after treatment. Analysis revealed mean area-under-the-curve (AUC) serum Δ4 concentrations to be higher during Δ4 treatment [2177 (SEM 100) nmol · l⁻¹] than Δ4Diol [900 (SEM 96) nmol · l⁻¹] or PL [484 (SEM 82) nmol · l⁻¹; P < 0.0001]. The Δ4 treatment also revealed a significant effect on total testosterone with a mean AUC [1632.5 (SEM 121) nmol · l⁻¹] that was greater than PL [1418.5 (SEM 131) nmol · l⁻¹; P < 0.05] but not significantly different from those observed after Δ4Diol treatment [1602.9 (SEM 119) nmol · l⁻¹; P = 0.77]. Free testosterone concentrations followed a similar pattern where mean AUC for the Δ4 treatment [6114.0 (SEM 600) pmol · l⁻¹] was greater than after PL [4974.6 (SEM 565) pmol · l⁻¹; P < 0.06] but not significantly different from those observed after Δ4Diol [5632.0 (SEM 389) pmol · l⁻¹; P = 0.48]. The appearance and apparent conversion to total and free testosterone over 90 min was stronger for the Δ4 treatment (r = 0.91, P < 0.045) than for Δ4Diol treatment (r = 0.69, NS) and negatively correlated for PL (r = −0.90, P < 0.02). These results would suggest that Δ4, and perhaps Δ4Diol, taken by month are capable of producing in vivo increases in testosterone concentrations in apparently healthy young men as has already been observed in women after treatment with Δ4. Accepted: 26 August 1999     

Ion channels in isolated mouse jejunal crypts

 The patch-clamp technique was used to characterise the ion channels in cells located in the mid region of mouse jejunal crypts. Six different channels were seen. A large outwardly rectified K⁺ channel (BK) (conductance, g at 0 mV = 92 ± 6 pS), which was highly selective for K⁺ [P _K ⁺ (1) > P _Rb ⁺ (0.6) >> P _Cs ⁺ (0.09) ≈ P _Na ⁺ (0.07) > P _Li ⁺ (0.04)], had a low, voltage-independent open probability (P _o) in the on-cell (O/C) configuration and appeared in 66% of the patches. In inside-out (I/O) patches, this channel had a linear current/voltage (I/V) relationship (g = 132 ± 3 pS), P _o was voltage dependent and it was blocked by cytoplasmic Ba²⁺ (5 mmol/l). An intermediate K⁺ channel (IK) which was present in 49% of O/C patches, had a linear I/V (g = 38 ± 3 pS), ran-down in O/C patches, and was not seen in I/O patches. A number of smaller channels (SC) with conductances ranging from 5 to 20 pS were seen in 16% of O/C patches. Also present in the basolateral membrane were a Cl^– channel (ICOR) and a nonselective cation channel (NSCC). These channels were only seen in I/O patches. ICOR had an outwardly rectified conductance (g at 0 mV = 36 ± 2 pS), its P _o was independent of voltage and unaffected by variations in cytoplasmic Ca²⁺ (100 nmol/l to 1 mmol/l) or ATP (0–1 mmol/l). The NSCC had a linear conductance (20 ± 1 pS), its P _o increased with depolarisation and elevation of cytoplasmic [Ca²⁺] (≥ 10 μmol/l), but was reduced by cytoplasmic ATP. None of the basolateral channels described here were activated by cAMP-dependent secretagogues, although a Cl^– conductance was activated. This cAMP-dependent Cl^– conductance was distinct from the basolateral Cl^– channel and thus is most likely located in the apical membrane. Received: 25 June 1997 / Accepted: 14 October 1997     

Characterization of an outwardly rectifying chloride channel in a human submandibular gland duct cell line (HSG)

We have used the single-channel patch-clamp technique to study ion channels in the plasma membrane of the HSG human submandibular gland duct cell line. In cell-attached and excised inside-out patches, at least six channel types were observed. When the pipette contained an isotonic KCl-rich solution and the bath an isotonic NaCl-rich solution, the predominant channel type seen in excised inside-out patches was a Cl⁻ channel with an outwardly rectifying current/voltage (I/V) relation that had a conductance of 12 pS at positive pipette potentials and 43 pS at negative pipette potentials. The channel was only seen in excised patches and its open probability was not significantly increased by membrane depolarization. The channel selectivity sequence (relative to Cl⁻) was estimated from reversal potential measurements to be: SCN⁻ (1.8)>NO ₃ ⁻ (1.4)> I⁻ (1.1) ∼ Cl⁻ (1) ∼ Br⁻ (0.8) > acetate (0.35). In inside-out patches the channel was blocked by addition of 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB) (100 μmol/l) to the bath but not by 9-anthracene carboxylic acid (9-AC) (100 μmol/l). The channel was not activated by increases in the free Ca²⁺ concentration on the cytosolic surface. This is the first report of an outwardly rectifying Cl⁻ channel in a salivary epithelium. The properties of this channel are not in accordance with the properties of the Cl⁻ conductances in the acinar or duct tissues which have been studied so far and its physiological role is unclear.     

A stretch-activated K+ channel in the basolateral membrane ofXenopus kidney proximal tubule cells

The present study examined whether a basolateral potassium ion (K⁺) channel is activated by membrane-stretching in the cell-attached patch. A K⁺ channel of conductance of 27.5 pS was most commonly observed in the basolateral membrane ofXenopus kidney proximal tubule cells. Channel activity increased with hyperpolarizing membrane potentials [at more positive pipette potentials (V _p)]. Open probability (P _o) was 0.03, 0.13, and 0.21 atV _p values of 0, 40, and 80 mV, respectively. Barium (0.1 mM) in the pipette reducedP _o by 79% at aV _p of 40 mV. Application of negative hydraulic pressure (−16 to −32 cm H₂O) to the pipette markedly activated outward currents (fromP _o=0.01 to 0.75) at aV _p of −80 mV, but not inward currents at aV _p of 80 mV. The size of the activated outward currents (from cell to pipette) did not change by replacing chloride with gluconate in the pipette. These results indicate that a stretch-activated K⁺ channel exists in the basolateral membrane of proximal tubule cells. It may play an important role as a K⁺ exit pathway when the cell membrane is stretched (for example, by cell swelling).     

Association of recreational physical activity with homocysteine,folate and lipid markers in young women

We assessed the influence of recreational physical activity in young healthy women on homocysteine, a potential risk factor for cardiovascular disease (CVD). Participants were 124 23-year-old normal-weight Italian recreational athletes (performing 8.7 ± 2.46 h week⁻¹ exercise) and 116 controls. Median blood homocysteine, folate and lipid markers did not differ between athletes and controls. Elevated homocysteine levels at CVD risk ≥12.0 and ≥15.0 μmol l⁻¹ were not different between groups. Continuous homocysteine was inversely related to folate (P < 0.001), positively associated with age (P = 0.009) and creatinine (P = 0.033), but not associated with hours of exercise, body mass index, and lipid markers. Women with folate depletion (<3.0 μg l⁻¹) were 4.5-fold more likely to have homocysteine ≥15.0 μmol l⁻¹. Recreational physical exercise does not adversely impact homocysteine levels among young women. Only low folate significantly increases the risk for hyperhomocysteinemia in young women.     

Physiological and metabolic responses of female games and endurance athletes to prolonged, intermittent, high-intensity running at 30° and 16°C ambient temperatures

Eight female games players (GP) and eight female endurance athletes (EA) ran intermittently at high-intensity and for prolonged periods in hot (30°C) and moderate (16°C) ambient temperatures. The subjects performed a two-part (A, B) test based on repeated 20-m shuttle runs. Part A comprised 60 m of walking, a maximal 15-m sprint, 60 m of cruising (90% maximal oxygen uptake, V˙O_2max) and 60 m of jogging (45% V˙O_2max) repeated for 75 min with a 3-min rest every 15 min. Part B involved an exercise and rest pattern of 60-s running at 100% V˙O_2max and 60-s rest which was continued until fatigue. Although the GP and EA did not respond differently in terms of distances completed, performance was 25 (SEM 4)% less (main effect trial, P < 0.01) in the hot (HT) compared with the moderate trial (MT). Sprints of 15 m took longer to complete in the heat (main effect, trial, P < 0.01), and sprint performance declined during HT but not MT (interaction, trial × time, P < 0.01). A very high correlation was found between the rate of rise in rectal temperature in HT and the distance completed [GP, r =−0.94, P < 0.01; EA (n = 7), r = −0.93, P < 0.01]. Blood lactate [La⁻ ]_b and plasma ammonia [NH₃]_p1 concentrations were higher for GP than EA, but were similar in HT and MT [La⁻ ]_b, HT: GP vs EA, 8.0 (SEM 0.9) vs 4.9 (SEM 1.1) mmol · l⁻¹; MT: GP vs EA, 8.0 (SEM 1.3) vs 4.4 (SEM 1.2) mmol · l⁻¹; interaction, group × time, P < 0.01; [NH₃]_p1, HT: GP vs EA, 70.1 (SEM 12.7) vs 43.2 (SEM 6.1) mmol · l⁻¹; MT: GP vs EA, 76.8 (SEM 8.8) vs 32.5 (SEM 3.8) μmol · l⁻¹; interaction, group × time, P < 0.01. Ad libitum water consumption was higher in HT [HT: GP vs EA, 18.9 (SEM 2.9) vs 13.5 (SEM 1.7) ml · kg⁻¹ · h⁻¹; MT: GP vs EA, 12.7 (SEM 3.7) vs 8.5 (SEM 1.5) ml · kg⁻¹ · h⁻¹; main effect, group, n.s.; main effect, trial, P < 0.01]. These results would suggest that elevated body temperature is probably the key factor limiting performance of prolonged, intermittent, high-intensity running when the ambient temperature is high, but not because of its effect on the metabolic responses to exercise. Accepted: 19 July 1999     

Ca2+ regulated K+ and non-selective cation channels in the basolateral membrane of rat colonic crypt base cells

 We have previously shown that a new type of K⁺ channel, present in the basolateral membrane of the colonic crypt base (blm), is necessary for cAMP-activated Cl^- secretion. Under basal conditions, and when stimulated by carbachol (CCH) alone, this channel is absent. In the present patch clamp-study we examined the ion channels present in the blm under cell-attached and in cell-excised conditions. In cell-attached recordings with NaCl-type solution in the pipette we measured activity of a K⁺ channel of 16 ± 0.3 pS (n = 168). The activity of this channel was sharply increased by CCH (0.1 mmol/l, n = 26). Reduction of extracellular Ca²⁺ to 0.1 mmol/l (n = 34) led to a reversible reduction of activity of this small channel (SK_Ca). It was also inactivated by forskolin (5 μmol/l, n = 38), whilst the K⁺ channel noise caused by the very small K⁺ channel increased. Activity of non-selective cation channels (NS_cat) was rarely observed immediately prior to the loss of attached basolateral patches and routinely in excised patches. The NS_cat, with a mean conductance of 49 ± 1.0 pS (n = 96), was Ca²⁺ activated and required >10 μmol/l Ca²⁺ (cytosolic side = cs). It was reversibly inhibited by ATP (<1 mmol/l, n = 13) and by 3′,5-dichloro-diphenylamine-2-carboxylate (10–100 μmol/l, n = 5). SK_Ca was also Ca²⁺ dependent in excised inside-out basolateral patches. Its activity stayed almost unaltered down to 1 μmol/l (cs) and then fell sharply to almost zero at 0.1 μmol/l Ca²⁺ (cs, n = 12). SK_Ca was inhibited by Ba²⁺ (n = 31) and was charybdotoxin sensitive (1 nmol/l) in outside-out basolateral patches (n = 3). Measurements of the Ca²⁺ activity ([Ca²⁺]_i) in these cells using fura-2 indicated that forskolin and depolarization, induced by an increase in bath K⁺ concentration to 30 mmol/l, reduced [Ca²⁺]_i markedly (n = 8–10). Hyperpolarization had the opposite effect. The present data indicate that the blm of these cells contains a small-conductance Ca²⁺-sensitive K⁺ channel. This channel is activated promptly by very small increments in [Ca²⁺]_i and is inactivated by a fall in [Ca²⁺]_i induced by forskolin. Received: 15 April 1996 / Received after revision and accepted: 17 June 1996     

Exercise training induces similar elevations in the activity of oxoglutarate dehydrogenase and peak oxygen uptake in the human quadriceps muscle

During exercise involving a small muscle mass, peak oxygen uptake is thought to be limited by peripheral factors, such as the degree of oxygen extraction from the blood and/or mitochondrial oxidative capacity. Previously, the maximal activity of the Krebs cycle enzyme oxoglutarate dehydrogenase has been shown to provide a quantitative measure of maximal oxidative metabolism, but it is not known whether the increase in this activity after a period of training reflects the elevation in peak oxygen consumption. Fourteen subjects performed one-legged knee extension exercise for 5–7 weeks, while the other leg remained untrained. Thereafter, the peak oxygen uptake by the quadriceps muscle was determined for both legs, and muscle biopsies were taken for assays of maximal enzyme activities (at 25°C). The peak oxygen uptake was 26% higher in the trained than in the untrained muscle (395 vs. 315 ml min⁻¹ kg⁻¹, respectively; P < 0.01). The maximal activities of the Krebs cycle enzymes in the trained and untrained muscle were as follows: citrate synthase, 22.4 vs. 18.2 μmol min⁻¹ g⁻¹ (23%, P < 0.05); oxoglutarate dehydrogenase, 1.88 vs. 1.54 μmol min⁻¹ g⁻¹ (22%, P < 0.05); and succinate dehydrogenase, 3.88 vs. 3.28 μmol min⁻¹ g⁻¹ (18%, P < 0.05). The difference between the trained and untrained muscles with respect to peak oxygen uptake (80 ml min⁻¹ kg⁻¹) corresponded to a flux through the Krebs cycle of 1.05 μmol min⁻¹ g⁻¹, and the corresponding difference in oxoglutarate dehydrogenase activity (at 38°C) was 0.83 μmol min⁻¹ g⁻¹. These parallel increases suggest that there is no excess mitochondrial capacity during maximal exercise with a small muscle mass.     

Muscle triacylglycerol and hormone-sensitive lipase activity in untrained and trained human muscles

During exercise, triacylglycerol (TG) is recruited in skeletal muscles. We hypothesized that both muscle hormone-sensitive lipase (HSL) activity and TG recruitment would be higher in trained than in untrained subjects in response to prolonged exercise. Healthy male subjects (26 ± 1 years, body moss index 23.3 ± 0.5 kg m⁻²), either untrained (N = 8, VO_2max 3.8 ± 0.2 l min⁻¹) or trained (N = 8, VO_2max 5.1 ± 0.1 l min⁻¹), were studied. Before and after 3-h exercise (58 ± 1% VO_2max), a biopsy was taken. Muscle citrate synthase (32 ± 2 vs. 47 ± 6 μmol g⁻¹ min⁻¹ d.w.) and β-hydroxy-acyl-CoA-dehydrogenase (38 ± 3 vs. 52 ± 5 μmol g⁻¹ min⁻¹ d.w.) activities were lower in untrained than in trained subjects (p < 0.05). Throughout the exercise, fat oxidation was higher in trained than in untrained subjects (p < 0.05). Muscle HSL activity was similar at rest (0.72 ± 0.08 and 0.74 ± 0.03 mU mg⁻¹ protein) and after exercise (0.71 ± 0.1 and 0.68 ± 0.03 mU mg⁻¹ protein) in untrained and trained subjects. At rest, the chemically determined muscle TG content (37 ± 8 and 26 ± 5 mmol g⁻¹ d.w.) was similar (p > 0.05), and after exercise it was unchanged in untrained and lower (p < 0.05) in trained subjects (41 ± 9 and 10 ± 2 mmol g⁽¹ d.w.). Determined histochemically, TG was decreased (p < 0.05) after exercise in type I and II fibres. Depletion of TG was not different between fibre types in untrained, but tended to be higher (p = 0.07) in type I compared with type II fibres in trained muscles. In conclusion, HSL activity is similar in untrained and trained skeletal muscles both before and after prolonged exercise. However, the tendency to higher muscle TG recruitment during exercise in the trained subjects suggests a difference in the regulation of HSL or other lipases during exercise in trained compared with untrained subjects.     

Aldosterone-induced changes in the cardiac L-type Ca2+ current can be prevented by antioxidants in vitro and are absent in rats on low salt diet

Mineralocorticoid receptor (MR) activation modulates cardiac L-type Ca²⁺ current (I _CaL) and transient outward K⁺ current (I _to). The exact circumstances of MR activation, however, remain elusive. Here, we investigate the influence of corticosteroids on MR-mediated changes in cellular electrophysiology. In vitro incubation of adult rat ventricular myocytes with the MR agonist aldosterone (100 nM, 24 h) increased I _CaL density by 34% (n = 16; p < 0.01). This effect was abrogated by co-incubation with the MR antagonist spironolactone (10 μM). To investigate whether an increase in serum aldosterone concentration is sufficient for an increase in I _CaL in vivo, rats were subjected to low Na⁺ diet (LSD, 0.013% Na⁺) for 28 days. This increased serum aldosterone concentration from 0.19 ± 0.04 nM (n = 6) in control animals (0.3% Na⁺, CSD) to 16.1 ± 2.1 nM (n = 6; p < 0.0001). Strikingly, I _CaL density was similar in both CSD and LSD rats (−12.9 ± 0.9 pA pF⁻¹, n = 18 and −13.7 ± 1.1 pA pF⁻¹, n = 16, respectively), as was I _to density. In vitro, the glucocorticoid corticosterone (1 μM) also increased I _CaL and this effect was blocked by spironolactone (10 μM). Co-incubation with corticosterone (1 μM, the normal serum concentration) and aldosterone (100 nM, mimicking low Na⁺ intake) did not further increase I _CaL compared to corticosterone alone. Moreover, co-incubation of myocytes with N-acetylcysteine (10 mM) prevented the aldosterone (100 nM) or corticosterone (1 μM)-induced increase in I _CaL. In conclusion, an increase in serum aldosterone concentration in response to LSD is not sufficient for an increase in I _CaL density in cardiomyocytes in vivo. This is supported in vitro by the absence of an effect of aldosterone on I _CaL in the presence of a physiological concentration of corticosterone. Moreover, the cellular redox state may modulate MR activation. Michael Wagner and Elena Rudakova contributed equally to this work.     

Energy cost and mechanical efficiency of riding a four-wheeled, human-powered, recumbent vehicle

Oxygen consumption at steady state (V˙O ₂, l · min⁻¹) and mechanical power (W˙, W) were measured in five subjects riding a human-powered vehicle (HPV, the Karbyk, a four-wheeled recumbent cycle) on a flat concrete road at constant sub-maximal speeds. The external mechanical work spent per unit of distance (W, J · m⁻¹), as calculated from the ratio of W˙ to the speed (v, m · s⁻¹), was found to increase with the square of v: W˙=8.12+(0.262 ·v ²) (r=0.986, n=31), where the first term represents the mechanical energy wasted, over a unit of distance, against frictional forces (rolling resistance, R_r), and the second term (k · v ²) is the work performed, per unit distance, to overcome the air drag. The rolling coefficient (C_r, obtained dividing R_r by m · g, where m is the overall mass and g is the acceleration of gravity) amounted to [mean (SD)] 0.0084 (0.0008), that is about 60% higher than that of a racing bicycle. The drag coefficient was calculated from the measured values of k, air density (ρ) and frontal area (A) [C_x=k · (0.5 · A · ρ)⁻¹], and amounted to 1.067 (0.029), that is about 20% higher than that of a racing bicycle. The energy cost of riding the HPV (C_k, J · m⁻¹) was measured from the ratio of metabolic power above rest (net V˙O ₂, expressed in J · s⁻¹) to the speed (v, m · s⁻¹); the value of this parameter increased with the square of v, as described by: C_k=61.45 + (0.675 · v ²) (r=0.711, n=23). The net mechanical efficiency (η) was calculated from the ratio of W to C_k: over the investigated speed range this turned out to be 0.22 (0.021). Best performance times (BPTs) of a “typical”élite athlete riding the Karbyk were calculated over the distances of 1, 5 and 10 km: these were about 8% longer than the BPTs calculated, on the same subjects, when riding a conventional racing bicycle. Accepted: 7 August 2000     

Effects of repeated muscle contractions on the tendon structures in humans

The purpose of this study was to investigate the changes in the elastic properties of tendons in humans in relation to fatigue of knee extensor muscles. The muscle fatigue test (MFT) consisted of maximal isometric contractions performed 50 times. The decline in peak moment was 43.6 (SD 19.5)%. After MFT, the muscle thickness and pennation angle of the vastus lateralis muscle (VL) significantly increased 1.5 (SD 0.7) mm (5%) and 1.7 (SD 1.8)° (11%), respectively. Before and after MFT, the elongation (l) of the tendon and aponeurosis of VL was directly measured by ultrasonography, while the subjects performed ramp isometric knee extensions up to maximal voluntary contraction . The l tended to be greater after MFT than before MFT. This difference in the l was statistically significant (P < 0.05) at force developments beyond 220 N. Furthermore, the compliance increased significantly from 2.0 (SD 0.6) · 10⁻² mm · N⁻¹ before MFT to 2.6 (SD 0.7) · 10⁻² mm · N⁻¹ after MFT (22.7%). In addition, the electromechanical delay was significantly increased from 60.6 (SD 5.9) ms before to 70.0 (SD 4.4) ms after MFT. These results suggested that the repeated muscle contractions made the tendon structures more compliant. Accepted: 15 August 2000     

Extracellular pH defense against lactic acid in normoxia and hypoxia before and after a Himalayan expedition

The extracellular pH defense against the lactic acidosis resulting from exercise can be estimated from the ratios −Δ[La] · ΔpH⁻¹ (where Δ[La] is change in lactic acid concentration and ΔpH is change in pH) and Δ[HCO₃ ⁻] · ΔpH⁻¹ (where Δ[HCO₃ ⁻] is change in bicarbonate concentration) in blood plasma. The difference between −Δ[La] · ΔpH⁻¹ and Δ[HCO₃ ⁻] · ΔpH⁻¹ yields the capacity of available non-bicarbonate buffers (mainly hemoglobin). In turn, Δ[HCO₃ ⁻] · ΔpH⁻¹ can be separated into a pure bicarbonate buffering (as calculated at constant carbon dioxide tension) and a hyperventilation effect. These quantities were measured in 12 mountaineers during incremental exercise tests before, and 7–8 days (group 1) or 11–12 days (group 2) after their return from a Himalayan expedition (2800–7600 m altitude) under conditions of normoxia and acute hypoxia. In normoxia −Δ[La] · ΔpH⁻¹ amounted to [mean (SEM)] 92 (6) mmol · l⁻¹ before altitude, of which 19 (4), 48 (1) and 25 (3) mmol · l⁻¹ were due to hyperventilation, bicarbonate and non-bicarbonate buffering, respectively. After altitude −Δ[La] · ΔpH⁻¹ was increased to 128 (12) mmol · l⁻¹ (P < 0.01) in group 1 and decreased to 72 (5) mmol · l⁻¹ in group 2 (P < 0.05), resulting mainly from apparent large changes of non-bicarbonate buffer capacity, which amounted to 49 (14) mmol · l⁻¹ in group 1 and to 10 (2) mmol · l⁻¹ in group 2. In acute hypoxia the apparent increase in non-bicarbonate buffers of group 1 was even larger [140 (18) mmol · l⁻¹]. Since the hemoglobin mass was only modestly elevated after descent, other factors must play a role. It is proposed here that the transport of La⁻ and H⁺ across cell membranes is differently influenced by high-altitude acclimatization. Accepted: 14 September 2000     

Gender differences in sweat lactate

Sweat rate may affect sweat lactate concentration. The current study examined potential gender differences in sweat lactate concentrations because of varying sweat rates. Males (n=6) and females (n=6) of similar age, percentage body fat, and maximal oxygen consumption (VO_2max) completed constant load (CON) cycling (30 min – approximately 40% VO_2max) and interval cycling (INT) (15 1-min intervals each separated by 1 min of rest) trials at 32 (1) °C wet bulb globe temperature (WBGT). Trials were preceded by 15 min of warm-up (0.5 kp, 60 rpms) and followed by 15 min of rest. Blood and sweat samples were collected at 15, 25, 35, 45, and 60 min during each trial. Total body water loss was used to calculate sweat rate. Blood lactate concentrations (CON ≅ 2 mmol · l⁻¹, INT ≅ 6 mmol · l⁻¹) and sweat lactate concentrations (CON and INT ≅ 12 mmol · l⁻¹) were not significantly different (P > 0.05) at any time between genders for CON or INT. Overall sweat rates (ml · h⁻¹) were not significantly different (P > 0.05) between trials but were significantly greater (P ≤ 0.05) for males than for females for CON [779.7 (292.6) versus 450.3 (84.6) ml · h⁻¹] and INT [798.0 (268.3) versus 503.0 (41.4) ml · h⁻¹]. However, correcting for surface area diminished the difference [CON: 390.7 (134.4) versus 277.7 (44.4) ml · h⁻¹, INT: 401.5 (124.1) versus 310.6 (23.4) ml · h⁻¹ (P ≤ 0.07)]. Estimated total lactate secretion was significantly greater (P ≤ 0.05) in males for CON and INT. Results suggest that sweat rate differences do not affect sweat lactate concentrations between genders. Accepted: 7 February 2000