Meropenem Prevents Levofloxacin-Induced Resistance in Penicillin-Resistant Pneumococci and Acts Synergistically with Levofloxacin in Experimental Meningitis

The aim of the present study was to investigate the potential synergy between meropenem and levofloxacin in vitro and in experimental meningitis and to determine the effect of meropenem on levofloxacin-induced resistance in vitro. Meropenem increased the efficacy of levofloxacin against the penicillin-resistant pneumococcal strain KR4 in time-killing assays in vitro and acted synergistically against a second penicillin-resistant strain WB4. In the checkerboard, only an additive effect (FIC indices: 1.0) was observed for both strains. In cycling experiments in vitro, levofloxacin alone led to a 64-fold increase in the MIC for both strains after 12 cycles. Addition of meropenem in sub-MIC concentrations (0.25×MIC) completely inhibited the selection of levofloxacin-resistant mutants in WB4 after 12 cycles. In KR4, the addition of meropenem led to just a twofold increase in the MIC for levofloxacin after 12 cycles. Mutations detected in the genes encoding for topoisomerase IV (parC) and gyrase (gyrA) confirmed the levofloxacin-induced resistance in both strains. Addition of meropenem was able to completely suppress levofloxacin-induced mutations in WB4 and led to only one mutation in parE in KR4. In experimental meningitis, meropenem, given in two doses (2×125 mg/kg), produced a good bactericidal activity (–0.45 log₁₀ cfu/ml·h) comparable to one dose (1×10 mg/kg) of levofloxacin (–0.44 log₁₀ cfu/ml·h) against the penicillin-resistant strain WB4. Meropenem combined with levofloxacin acted synergistically (–0.93 log₁₀ cfu/ml·h), sterilizing the CSF of all rabbits.     

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     

Oxygen uptake kinetics during treadmill running across exercise intensity domains

The purpose of the present study was to examine comprehensively the kinetics of oxygen uptake ( ) during treadmill running across the moderate, heavy and severe exercise intensity domains. Nine subjects [mean (SD age, 27 (7) years; mass, 69.8 (9.0) kg; maximum , , 4,137 (697) ml·min^–1] performed a series of "square-wave" rest-to-exercise transitions of 6 min duration at running speeds equivalent to 80% and 100% of the at lactate threshold (LT; moderate exercise); and at 20%, 40%, 60%, 80% and 100% of the difference between the at LT and (Δ, heavy and severe exercise). Critical velocity (CV) was also determined using four maximal treadmill runs designed to result in exhaustion in 2–15 min. The response was modelled using non-linear regression techniques. As expected, the amplitude of the primary component increased with exercise intensity [from 1,868 (136) ml·min^–1 at 80% LT to 3,296 (218) ml·min^–1 at 100% Δ, P<0.05]. However, there was a non-significant trend for the "gain" of the primary component to decrease as exercise intensity increased [181 (7) ml·kg^–1·km^–1 at 80% LT to 160 (6) ml·kg^–1·km^–1 at 100% Δ]. The time constant of the primary component was not different between supra-LT running speeds (mean value range = 17.9–19.1 s), but was significantly shorter during the 80% LT trial [12.7 (1.4) s, P<0.05]. The slow component increased with exercise intensity from 139 (39) ml·min^–1 at 20% Δ to 487 (57) ml·min^–1 at 80% Δ (P<0.05), but decreased to 317 (84) ml·min^–1 during the 100% Δ trial (P<0.05). During both the 80% Δ and 100% Δ trials, the at the end of exercise reached [4,152 (242) ml·min^–1 and 4,154 (114) ml·min^–1, respectively]. Our results suggest that the "gain" of the primary component is not constant as exercise intensity increases across the moderate, heavy and severe domains of treadmill running. These intensity-dependent changes in the amplitudes and kinetics of the response profiles may be associated with the changing patterns of muscle fibre recruitment that occur as exercise intensity increases. Electronic Publication     

Effect of water ingestion on cardiovascular and thermal responses to prolonged cycling and running in humans: a comparison

The aim of this investigation was to examine the effect of water ingestion on physiological responses to prolonged cycling (CYC) and running (RUN). A group of 11 men with mean (SEM) maximal oxygen uptake (V˙O_2max) 48.5 (1.8) ml·kg^–1·min^–1 on a cycle-ergometer and 52.1 (2.2) ml·kg^–1·min^–1 on a treadmill (P<0.01) exercised for 90 min on four occasions, twice on each ergometer, at 60% of mode specific V˙O_2max. No fluid was taken (D) in one trial on each ergometer, whereas 60% of fluid losses were replaced by drinking water in the other trial (W). In CYC, water ingestion attenuated the change in cardiac output ( ) and the reduction in stroke volume (ΔSV) [ΔSV: –22.7 (3.8) in D, –10.7 (2.9) ml·beat^–1 in W, P<0.01; : –1.9 (0.5) in D, –0.2 (0.4) l·min^–1 in W at 85 min, P<0.01], but did not affect rectal temperature [T _re at 90 min: 38.8 (0.1)°C in D, 38.7 (0.1)°C in W]. In contrast, fluid replacement reduced hyperthermia in RUN [T _re at 90 min: 39.6 (0.2) in D, 39.1 (0.2)°C in W, P<0.01], and this was linked with a higher skin blood flow [RUN-W 88.9 (8.5), RUN-D 70.7 (8.4)%, P<0.05]. The and ΔSV were also attenuated with water ingestion in this mode of exercise (P<0.05). It is concluded that water ingestion improves physiological function in both cycling and running, but that the underlying mechanism is different in the two modes of exercise. Electronic Publication     

Efficacy of rifampin, in monotherapy and in combinations, in an experimental murine pneumonia model caused by panresistant Acinetobacter baumannii strains

The objective of this work was to evaluate the efficacy of rifampin, and its combinations with imipenem or sulbactam, in an experimental pneumonia model caused by two panresistant Acinetobacter baumannii strains (HUVR99 and HUVR113). Minimum inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs) (μg/ml) of the strains were rifampin 128/>128 for both strains, imipenem 128/>256 and 256/>256 for HUVR99 and HUVR113, respectively, and sulbactam >256/>256 for both strains. In time–kill studies, at MICs, rifampin was bactericidal for both strains and sulbactam against the HUVR99 strain. Rifampin plus imipenem or sulbactam, at the MIC or mice C _max, were synergistic. In vivo, against HUVR99 and HUVR113, rifampin (73% and 40%) and its combinations improved the survival with respect to the control group (20% and 0%, p < 0.05), respectively. Rifampin (87% and 46%) and its combinations improved the sterilization of blood cultures with respect to the control groups (0%, p < 0.05). In regard to the bacterial clearance from lungs, rifampin (2.57 ± 2.47 and 5.35 ± 3.03 log₁₀ cfu/g) and its combinations with imipenem or sulbactam diminished the bacterial lung concentration with respect to the control group (10.89 ± 3.00 and 11.86 ± 0.49, p < 0.05) with both strains. In conclusion, rifampin alone or associated to imipenem or sulbactam were effective for the treatment of murine pneumonia caused by selected panresistant A. baumannii strains.     

Losartan antagonism of angiotensin-II-induced potassium secretion across rat colon

 The effect of angiotensin II (ANG II) on potassium transport across the short-circuited rat distal colon was investigated using ⁸⁶Rb⁺ as a tracer for unidirectional K⁺ fluxes. The addition of high concentrations of ANG II (≥10^–6 M) to the serosal bathing solution had no effect on the mucosal to serosal flux of Rb⁺, but significantly increased the serosal to mucosal flux and abolished the basal net absorptive Rb⁺ flux. These ANG-II-induced alterations in Rb⁺ transport were blocked by the AT₁ receptor antagonist losartan and its metabolite EXP3174, which is also known to have AT₁ receptor antagonistic activity. In contrast, an AT₂ receptor antagonist, PD123319, did not prevent the alterations in colonic Rb⁺ transport induced by ANG II in vitro. At lower bath concentrations (10^–7 M to 10^–10 M ), ANG II had no measurable effects on Rb⁺ transport across this tissue but ANG II did have a bimodal effect on short-circuit current (I _sc), indicating additional effects on the electrogenic transport of other ions. Dose-dependent reductions in I _sc were observed between 10^–7 M (↓ΔI _sc=1.96±0.49 μEq·cm^–2·h^–1, n=6) and 10^–10 M (↓ΔI _sc=0.16±0.19 μEq·cm^–2·h^–1, n=7) ANG II, whereas I _sc was increased at the higher concentrations (↑ΔI _sc= 3.36±0.52 μEq·cm^–2·h^–1, n=7, at 10^–4 M). The ANG-II-induced increases and decreases in I _sc were both blocked by losartan but not by PD123319. These studies are the first to demonstrate an effect of ANG II on K⁺ transport across rat colon that is independent of aldosterone and mediated by AT₁ receptors. Received: 13 March 1998 Received after revision and accepted: 26 May 1998     

Blood flow to the brown adipose tissue of conscious young rabbits during hypoxia in cold and warm conditions

 Brown adipose tissue (BAT) non-shivering thermogenesis is stimulated by cold temperature and depressed by hypoxia. We investigated the extent to which changes in metabolic rate during cold and hypoxia, singly or combined, were accompanied by changes in BAT perfusion. One-month-old rabbits were instrumented for measurements of regional blood flow by the coloured microsphere technique. One group of rabbits was tested in warm (24 °C, n=17), and the other in cold (13 °C, n=9) conditions, first in normoxia (inspired oxygen concentration FIO₂ about 21%, arterial oxygen saturation S _aO₂ approximately 88%) followed by hypoxia (FIO₂ approximately 10%, S _aO₂ approximately 54%). In warm conditions, oxygen consumption (V·O₂, measured by an open-flow method) averaged 22 ml·kg^–1·min^–1 (STPD), and BAT blood flow 98 ml·100g^–1·min^–1. In hypoxia, V·O₂ dropped on average to 87%, whereas BAT flow dropped to 43% of the normoxic values. In the cold during normoxia, V·O₂ averaged 31 ml·kg^–1·min^–1 (STPD), and BAT blood flow was 155 ml·100g^–1·min^–1. In cold and hypoxia V·O₂ dropped to 19 ml·kg^–1·min^–1 (STPD) (i.e. 60% of the normoxic value), whereas BAT blood flow was not altered significantly (148 ml·100g^–1·min^–1). Hence, BAT blood flow decreased in hypoxia in absence of cold stimuli, whereas it remained high when hypoxia occurred during cold, despite the major drop in V·O₂. We conclude that cold is more important than hypoxia in determining BAT perfusion, and that changes in BAT blood flow are not a mechanism for the hypoxic control of V·O₂. Received: 24 June 1998 / Received after revision: 21 September 1998 / Accepted: 29 September 1998     

The contribution of refractoriness to arrhythmic substrate in hypokalemic Langendorff-perfused murine hearts

The clinical effects of hypokalemia including action potential prolongation and arrhythmogenicity suppressible by lidocaine were reproduced in hypokalemic (3.0 mM K⁺) Langendorff-perfused murine hearts before and after exposure to lidocaine (10 μM). Novel limiting criteria for local and transmural, epicardial, and endocardial re-excitation involving action potential duration (at 90% repolarization, APD₉₀), ventricular effective refractory period (VERP), and transmural conduction time (Δlatency), where appropriate, were applied to normokalemic (5.2 mM K⁺) and hypokalemic hearts. Hypokalemia increased epicardial APD₉₀ from 46.6 ± 1.2 to 53.1 ± 0.7 ms yet decreased epicardial VERP from 41 ± 4 to 29 ± 1 ms, left endocardial APD₉₀ unchanged (58.2 ± 3.7 to 56.9 ± 4.0 ms) yet decreased endocardial VERP from 48 ± 4 to 29 ± 2 ms, and left Δlatency unchanged (1.6 ± 1.4 to 1.1 ± 1.1 ms; eight normokalemic and five hypokalemic hearts). These findings precisely matched computational predictions based on previous reports of altered ion channel gating and membrane hyperpolarization. Hypokalemia thus shifted all re-excitation criteria in the positive direction. In contrast, hypokalemia spared epicardial APD₉₀ (54.8 ± 2.7 to 60.6 ± 2.7 ms), epicardial VERP (84 ± 5 to 81 ± 7 ms), endocardial APD₉₀ (56.6 ± 4.2 to 63.7 ± 6.4 ms), endocardial VERP (80 ± 2 to 84 ± 4 ms), and Δlatency (12.5 ± 6.2 to 7.6 ± 3.4 ms; five hearts in each case) in lidocaine-treated hearts. Exposure to lidocaine thus consistently shifted all re-excitation criteria in the negative direction, again precisely agreeing with the arrhythmogenic findings. In contrast, established analyses invoking transmural dispersion of repolarization failed to account for any of these findings. We thus establish novel, more general, criteria predictive of arrhythmogenicity that may be particularly useful where APD₉₀ might diverge sharply from VERP.     

Carbon monoxide inhibits hypoxic pulmonary vasoconstriction in rats by a cGMP-independent mechanism

 Hypoxia activates erythropoietin-producing cells, chemoreceptor cells of the carotid body and pulmonary artery smooth muscle cells (PSMC) with a comparable arterial PO₂ threshold of some 70 mmHg. The inhibition by CO of the hypoxic responses in the two former cell types has led to the proposal that a haemoprotein is involved in the detection of the PO₂ levels. Here, we report the effect of CO on the hypoxic pulmonary vasoconstriction (HPV). Pulmonary arterial pressure (PAP) was measured in an in situ, blood-perfused lung preparation. PAP in normoxia (20% O₂, 5% CO₂) was 15.2±1.8 mmHg, and hypoxia (2% O₂, 5% CO₂) produced a ΔPAP of 6.3±0.4 mmHg. Addition of 8% or 15% CO to the hypoxic gas mixture reduced the ΔPAP by 88.3±2.7% and 78.2±6.1% respectively. The same levels of CO did not affect normoxic PAP nor reduced the ΔPAP produced by angiotensin II. The effect of CO was studied after inhibition of the NO-cyclic guanosine monophosphate (cGMP) cascade with N-methyl-l-arginine (5·10^–5 M) or methylene blue (1.4·10^–4 M). It was found that both inhibitors more than doubled the hypoxic ΔPAP without altering the effectiveness of CO to inhibit the HPV. In in vitro experiments we verified the inhibition of guanylate cyclase by measuring the levels of cGMP in segments of the pulmonary artery. Cyclic GMP levels were 1.4±0.2 (normoxia), 2.5±0.3 (hypoxia) and 3.3±0.5 pmole/mg tissue (hypoxia plus 8% CO); sodium nitroprusside increased normoxic cGMP levels about fourfold. Methylene blue reduced cGMP levels to less than 10% in all cases, and abolished the differences among normoxic, hypoxic and hypoxic plus CO groups. It is concluded that CO inhibits HPV by a NO-cGMP independent mechanism and it is proposed that a haemoprotein could be involved in O₂-sensing in PSMC. Received: 17 March 1997 / Received after revision: 10 June 1997 / Accepted: 11 July 1997     

High-intensity exercise decreases muscle buffer capacity via a decrease in protein buffering in human skeletal muscle

We have previously reported an acute decrease in muscle buffer capacity (βm_{in vitro}) following high-intensity exercise. The aim of this study was to identify which muscle buffers are affected by acute exercise and the effects of exercise type and a training intervention on these changes. Whole muscle and non-protein βm_{in vitro} were measured in male endurance athletes (VO_2max = 59.8 ± 5.8 mL kg⁻¹ min⁻¹), and before and after training in male, team-sport athletes (VO_2max = 55.6 ± 5.5 mL kg⁻¹ min⁻¹). Biopsies were obtained at rest and immediately after either time-to-fatigue at 120% VO_2max (endurance athletes) or repeated sprints (team-sport athletes). High-intensity exercise was associated with a significant decrease in βm_{in vitro} in endurance-trained males (146 ± 9 to 138 ± 7 mmol H⁺·kg d.w.⁻¹·pH⁻¹), and in male team-sport athletes both before (139 ± 9 to 131 ± 7 mmol H⁺·kg d.w.⁻¹·pH⁻¹) and after training (152 ± 11 to 142 ± 9 mmol H⁺·kg d.w.⁻¹·pH⁻¹). There were no acute changes in non-protein buffering capacity. There was a significant increase in βm_{in vitro} following training, but this did not alter the post-exercise decrease in βm_{in vitro}. In conclusion, high-intensity exercise decreased βm_{in vitro} independent of exercise type or an interval-training intervention; this was largely explained by a decrease in protein buffering. These findings have important implications when examining training-induced changes in βm_{in vitro}. Resting and post-exercise muscle samples cannot be used interchangeably to determine βm_{in vitro}, and researchers must ensure that post-training measurements of βm_{in vitro} are not influenced by an acute decrease caused by the final training bout.     

Effect of a 1 year combined aerobic- and weight-training exercise programme on aerobic capacity and ventilatory threshold in patients suffering from coronary artery disease

Weight-training is recommended as a complement to conventional aerobic-training for most low to moderate risk patients suffering from coronary artery disease (CAD). The purpose of this study was to evaluate the effect of a 1 year exercise programme combining weight- and aerobic-training on peak oxygen uptake (VO_2,peak) and ventilatory threshold (VT). We studied 40 men suffering CAD who were divided into three groups: 14 subjects to weight-training plus aerobic-training [mean (SD] [combined exercise group, age 55 (10) years], 14 to aerobic-training only [aerobic-training group, age 57 (11) years], and 12 to a control group [standard care, age 57 (11) years]. A symptom-limited graded exercise test using the standard Bruce protocol was performed using a 12-lead electrocardiogram, and gas analysis techniques. Muscle strength was determined only in the combined exercise group using the one-repetition maximum method on each of eight weight exercises. Arm and leg strength increased by 21.9% and 27.8% respectively (P<0.0001) from pre to post-tests. The VO_2,peak did not differ between the combined and aerobic-training groups but their adjusted means were greater than those of the control group [39 (1.8) and 35.3 (1.8) compared to 26.2 (2.7) ml·kg^–1·min^–1 (P<0.001)]. The oxygen uptake at VT was higher in the combined group [24.7 (1.4) ml·kg^–1·min^–1] compared to aerobic [18.7 (1.4) ml·kg^–1·min^–1] and control [13.6 (1.7) ml·kg^–1min^–1] groups (P<0.001). Similar results were found for exercise tolerance (treadmill time to peak and at VT). Combined exercise training increased the VT more than aerobic-training alone. Combined exercise training did not improve the VO_2,peak or the functional capacity more than aerobic-training alone. Electronic Publication     

Breath-by-breath fluctuations of pulmonary gas exchange and ventilation in COPD patients

The purpose of the study was to characterise statistically the inherent fluctuations in breath-by-breath measurements of pulmonary gas exchange (oxygen uptake and carbon dioxide output, VO₂ and VCO₂, respectively) and pulmonary ventilation (V_E) in patients with chronic obstructive pulmonary disease (COPD) and to compare them with those of healthy control subjects. Thirty subjects with COPD [mean (SD): 67 (6) years old; forced expiratory volume in 1 min, FEV₁ 1.25( 0.18) l; 42 (6)% predicted FEV₁] and 12 healthy subjects [31 (3) years old; FEV₁ 3.62 (0.54) l; 99 (8)% predicted FEV₁] performed exercise tests on a cycle ergometer at a constant work rate of moderate intensity. Steady-state exercise values for VO₂, VCO₂ and V_E were 905 (96) ml·min^–1, 847(90) ml·min^–1 and 23 (3) l·min^–1, respectively for the COPD patients and 1239(89) ml·min^–1, 1191(84) ml·min^–1 and 37(3) l·min^–1, respectively, for the healthy controls. The breath-by-breath fluctuations were well characterised by a Gaussian density-probability function with breath-to-breath autocorrelations that were not significantly different from 0, up to four subsequent breaths. Its magnitude varied among variables, but was independent of the signal amplitude for the same subject and variable. With ratios of amplitude of fluctuation:signal of around 10%, typical of the patients studied, the resolution of time constants and amplitude were ≅9 s and ≅100 ml·min^–1, respectively for VO₂ or VCO₂ with one repetition. Electronic Publication     

Infestation status of gnathiid isopod juveniles parasitic on Dusky grouper (<Emphasis Type="Italic">Epinephelus marginatus</Emphasis>) from the northeast Mediterranean Sea

This is the first detailed documented record of Gnathiid isopod praniza larvae infestating dusky grouper, (Epinephelus marginatus Lowe 1834) in the northeast Mediterranean Sea (36°36′N–36°07′E, 35°52′N–36°25′E). Fish were sampled monthly from Iskenderun Bay during a 3-year period from 2000 to 2003 [N = 468, W ± SD (range) = 503.69 ± 342.35 g (177–2,832 g), TL ± SD (range) = 32.39 ± 9.22 cm (16.1–67.0 cm), W _total = 0.213L _total ^2.19, r _total ² = 0.85]. Juveniles of the Gnathia sp. were only extracted from the epithelium of the buccal cavity. The monthly and seasonal patterns in infestation rates (mean prevalence, P = 27.35% and mean intensity, MI ± SD = 21.35 ± 16.19), and the relationship between length–weight and infested/non-infested fish were calculated. This study suggests that gnathiid parasite has no effect on the growth and general health condition of infested fish, although high intensities were observed in fish.     

Substrate utilization in boys during exercise with [13C]-glucose ingestion

The influence of glucose ingestion on substrate utilization during prolonged exercise in children and adolescents is currently unknown. In the present study we determined the effect of intermittent exogenous glucose (GLU_exo) ingestion on substrate utilization during prolonged exercise, in adolescent boys ages 13–17 years. Healthy untrained volunteers performed four 30-min exercise bouts on a cycle ergometer, separated by 5-min rest periods (≅60% maximum O₂ consumption), on two occasions spaced 1–4 weeks apart. Two trials were performed, a control trial (CT), in which subjects ingested water intermittently during the exercise, and a glucose trial (GT), in which subjects ingested a ¹³C-enriched GLU_exo drink (≅3 g glucose · kg body mass⁻¹), also intermittently during the exercise. Total free fatty acids (FAT_total), glucose (GLU_total) and carbohydrate (CHO_total) oxidation was determined from indirect calorimetry, while GLU_exo oxidation was calculated from the ¹³C/¹²C ratio in expired air after 5–10 min and 25–30 min of exercise in each bout. Heart rate and rating of perceived exertion (RPE) were determined at the same time intervals. The oxidation of CHO_total was 169.1 (12.9) g · 120 min⁻¹ and 203.1 (15.9) g · 120 min⁻¹ (P < 0.01) and that of FAT_total was 31.0 (4.2) g · 120 min⁻¹ and 17.1 (2.5) g · 120 min⁻¹ (P < 0.01) in CT and GT, respectively. GLU_exo oxidation in GT was 57.8 (4.3) g · 120 min⁻¹, or 34.2 (2.2)% of that ingested. Endogenous glucose oxidation was 169.1 (12.9) g · 120 min⁻¹ and 145.3 (11.9) g · 120 min⁻¹ (P < 0.01) in CT and GT, respectively. Insulin and glucose concentrations were higher in GT than in CT by 226% and 37%, respectively (both P < 0.05). Free fatty acids and glycerol concentrations were lower in GT than in CT, by 27% and 79%, respectively (both P < 0.05). Heart rate was similar between trials, but RPE was lower in GT vs CT at both 115 and 135 min. Thus, under these experimental conditions, GLU_exo intake spares endogenous carbohydrate and fat by 16% and 45%, respectively, contributes to approximately 25% of the total energy demand of exercise, and lowers the RPE. Accepted: 21 May 2000     

Activity of ethanol and daptomycin lock on biofilm generated by an in vitro dynamic model using real subcutaneous injection ports

Vancomycin lock solution (LS) is recommended for the conservative treatment of subcutaneous injection port (SIP)-related infections, but may be associated with failure. We used an in vitro dynamic model of biofilm formation in an SIP, based on a continuous flow circulating via a real SIP, to assess the effectiveness of vancomycin (5 mg/ml), daptomycin (5 mg/ml) and ethanol 40 % LS in eradicating a pre-established Staphylococcus epidermidis biofilm. Heparin, Ringer’s lactate and enoxaparin sodium LS were used as controls. The logarithmic reductions of colony-forming units (CFU) were compared by Student’s t-test. After 24 h of exposure, the vancomycin LS did not exert a greater bactericidal effect than the heparin LS control (mean logarithmic reduction: 2.27?±?0.58 vs. 1.34?±?0.22, respectively, p?=?0.3). The mean logarithmic reduction was greater with daptomycin LS (5.45?±?0.14 vs. 0.39?±?0.12, p?<?0.01) and ethanol LS (6.79?±?1.03 vs. 1.43?±?0.54, p?=?0.02). Bacterial revival after exposure to 24 h of LS was assessed. The mean viable bacteria count was significantly higher for vancomycin LS (9.36?±?0.10 log₁₀CFU) and daptomycin LS (9.16?±?0.02 log₁₀CFU) than for ethanol LS (2.95?±?1.65 log₁₀CFU). Ethanol appeared to be the most attractive option to treat SIP-related infection, but its poor ability to entirely disrupt the biofilm structure may require its use in association with a dispersal agent to avoid renewal of the biofilm.     

Sympathetic control of the forearm blood flow in man during brief isometric contractions

Summary Experiments were performed to assess the possible neurally mediated constriction in active skeletal muscle during isometric hand-grip contractions. Forearm blood flow was measured by venous occlusion plethysmography on 5 volunteers who exerted a series of repeated contractions of 4 s duration every 12 s at 60% of their maximum strength of fatigue. The blood flows increased initially, but then remained constant at 20–24 ml·min⁻¹·100 ml⁻¹ throughout the exercise even though mean arterial blood pressure reached 21–23 kPa (160–170 mm Hg). When the same exercise was performed after arterial infusion of phentolamine, forearm blood flow increased steadily to near maximal levels of 38.7±1.4 ml·min⁻¹·100 ml⁻¹. Venous catecholamines, principally norepinephrine, increased throughout exercise, reaching peak values of 983±258 pg·ml⁻¹ at fatigue. Of the vasoactive substances measured, the concentration of K⁺ and osmolarity in venous plasma also increased initially and reached a steady-state during the exercise but ATP increased steadily throughout the exercise. These data indicate a continually increasing α-adrenergic constriction to the vascular beds in active muscles in the human forearm during isometric exercise, that is only partially counteracted by vasoactive metabolites.     

Epac activation, altered calcium homeostasis and ventricular arrhythmogenesis in the murine heart

The recently described exchange protein directly activated by cAMP (Epac) has been implicated in distinct protein kinase A-independent cellular signalling pathways. We investigated the role of Epac activation in adrenergically mediated ventricular arrhythmogenesis. In contrast to observations in control conditions (n = 20), monophasic action potentials recorded in 2 of 10 intrinsically beating and 5 of 20 extrinsically paced Langendorff-perfused wild-type murine hearts perfused with the Epac activator 8-pCPT-2′-O-Me-cAMP (8-CPT, 1 μM) showed spontaneous triggered activity. Three of 20 such extrinsically paced hearts showed spontaneous ventricular tachycardia (VT). Programmed electrical stimulation provoked VT in 10 of 20 similarly treated hearts (P < 0.001; n = 20). However, there were no statistically significant accompanying changes (P > 0.05) in left ventricular epicardial (40.7 ± 1.2 versus 44.0 ± 1.7 ms; n = 10) or endocardial action potential durations (APD₉₀; 51.8 ± 2.3 versus 51.9 ± 2.2 ms; n = 10), transmural (ΔAPD₉₀) (11.1 ± 2.6 versus 7.9 ± 2.8 ms; n = 10) or apico-basal repolarisation gradients, ventricular effective refractory periods (29.1 ± 1.7 versus 31.2 ± 2.4 ms in control and 8-CPT-treated hearts, respectively; n = 10) and APD₉₀ restitution characteristics. Nevertheless, fluorescence imaging of cytosolic Ca²⁺ levels demonstrated abnormal Ca²⁺ homeostasis in paced and resting isolated ventricular myocytes. Epac activation using isoproterenol in the presence of H-89 was also arrhythmogenic and similarly altered cellular Ca²⁺ homeostasis. Epac-dependent effects were reduced by Ca²⁺/calmodulin-dependent protein kinase II (CaMKII) inhibition with 1 μM KN-93. These findings associate VT in an intact cardiac preparation with altered cellular Ca²⁺ homeostasis and Epac activation for the first time, in the absence of altered repolarisation gradients previously implicated in reentrant arrhythmias through a mechanism dependent on CaMKII activity.     

Mechanisms of reduced mitochondrial Ca2+ accumulation in failing hamster heart

Mitochondrial Ca²⁺ plays important roles in the regulation of energy metabolism and cellular Ca²⁺ homeostasis. In this study, we characterized mitochondrial Ca²⁺ accumulation in Syrian hamster hearts with hereditary cardiomyopathy (strain BIO 14.6). Exposure of isolated mitochondria from 70 nM to 30 μM Ca²⁺ ([Ca²⁺]_o) caused a concentration-dependent increase in intramitochondrial Ca²⁺ concentrations ([Ca²⁺]_m). The [Ca²⁺]_m was significantly lower in cardiomyopathic (CMP) hamsters than in healthy hamsters when [Ca²⁺]_o was higher than 1 μM and a decrease of about 52% was detected at [Ca²⁺]_o of 30 μM (916 ± 67 nM vs 1,932 ± 132 nM in control). A possible mechanism responsible for the decreased mitochondrial Ca²⁺ uptake in CMP hamsters is the depolarization of mitochondrial membrane potential (Δψ _m). Using a tetraphenylphosphonium (TPP⁺) electrode, the measured Δψ _m in failing heart mitochondria was −136 ± 1.5 mV compared with −159 ± 1.3 mV in controls. Analyses of mitochondrial respiratory chain demonstrated a significant impairment of complex I and complex IV activities in failing heart mitochondria. In summary, a less negative Δψ _m resulting from defects in the respiratory chain may lead to attenuated mitochondrial Ca²⁺ accumulation, which in turn may contribute to the depressed energy production and myocardial contractility in this model of heart failure. In addition to other known impairments of ion transport in sarcoplasmic reticulum and plasma membrane, results from this paper on mitochondrial dysfunctions expand our understanding of the molecular mechanisms leading to heart failure.     

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     

Evaluation of meropenem alone and combined with rifampin in the guinea pig model of pneumococcal meningitis

Meropenem is a broad-spectrum carbapenem antibiotic that is highly active against the pathogens causing meningitis. The aims of this study was to determine the efficacies of meropenem alone and combined with rifampin against two Streptococcus pneumoniae strains with different susceptibility to β-lactams using the guinea pig meningitis model and compare them with the standard ceftriaxone plus vancomycin therapy. All treatments except rifampin were bactericidal from 6 h. The addition of rifampin did not improve the activity of meropenem alone. Our results provide good evidence of the efficacy of meropenem in the treatment of penicillin- and cephalosporin-susceptible and -resistant pneumococcal meningitis similar to that of ceftriaxone plus vancomycin, suggesting that meropenem might be a good option in the management of this infection.