Phylogenetic analyses of the matrix and non-structural genes of equine influenza viruses

Summary.  Matrix (M) and nonstructural (NS) genes of thirteen equine H3N8 and H7N7 influenza viruses were sequenced and analyzed from an evolutionary point of view. The M and NS genes of H3N8 viruses isolated between 1989 and 1993 evolved into two minor branch clusters, including isolates from Europe and the American continent, respectively. It was noteworthy to reveal that the nucleotide sequences of the M and NS genes of an earlier American strain showed highest homology to those of recent European viruses. “Frozen evolution” was observed in the M and NS genes of A/eq/LaPlata/1/88. It was also evident that the NS gene of an H7N7 virus from 1977 was very similar to that of a 1979-H3N 8 virus, while the M gene was closest phylogenetically to that of the earliest H7N7 virus isolated in 1956. Furthermore, the M2 protein of A/eq/Newmarket/1/77 virus contained a carboxyl terminal deletion of three amino acids. The evolutionary rates of the M and NS genes of H3N8 equine influenza viruses were estimated to be 5.4 × 10⁻⁴ and 5.1 × 10⁻⁴ substitutions per site per year, respectively, which were slower than those of human viruses. Received November 21, 1997 Accepted March 9, 1998     

Effect of Lysophosphatidylcholine on α-Adrenoreactivity of Rat Aorta Smooth Muscles

In experiments on rat aortic ring segments, lysophosphatidylcholine in concentrations of 2 × 10⁻⁶, 2 × 10⁻⁵, and 2 × 10⁻⁴ M did not suppress the tonotropic effect of phenylephrine (6 × 10⁻⁶ and 6 × 10⁻⁵ M) and in concentration of 2 × 10⁻⁵ M even potentiated it, which was noted for phenylephrine at a concentration of 6 × 10⁻⁶ M. It was concluded that the chemomodulating effect of lysophosphatidylcholine depends on the type of receptors and cells.     

Apoptosis Inhibitor Expressed by Macrophages Tempers Autoimmune Colitis and the Risk of Colitis-Based Carcinogenesis in TCRα<Superscript>−/−</Superscript> Mice

Background Crohn’s disease and ulcerative colitis (UC) are the two main entities involved in human inflammatory bowel disease (IBD). However, their precise etiologies remain unclear. To study the development of mucosal inflammation, and chronic inflammation-based dysplasia and carcinoma formation, we examined possible roles of the apoptosis inhibitor expressed by macrophages (AIM) in an experimental IBD model. Methods In this study, we used T cell receptor α deficient (TCRα^−/−) mice, a known UC-like colitis model. We generated TCRα^−/− × AIM^−/− double knockout mice by crossbreeding TCRα^−/− with AIM^−/− mice. At 24 weeks of age, mice were killed to obtain colon tissues for pathological examinations. TCRα^−/− × AIM^+/− mice, heterozygous littermates of TCRα^−/− × AIM^−/− mice, were used as controls. Results Severe colitis was observed in TCRα^−/− × AIM^−/− mice, when compared with TCRα^−/− × AIM^+/− mice. Dysplasia was detected in TCRα^−/− × AIM^−/− mice, but not in TCRα^−/− × AIM^+/− mice. Adenocarcinoma formation was observed from dysplasia only in TCRα^−/− × AIM^−/− mice. Conclusion Not only a high incidence of severe colitis but also dysplasia and adenocarcinoma formation were observed in TCRα^−/− × AIM^−/− mice only. AIM have some regulatory roles in inflammation and progression of dysplasia to carcinoma in TCRα^−/− mice.     

Comparative evaluation of the sensitivity of LAMP, PCR and in vitro culture methods for the diagnosis of equine piroplasmosis

The sensitivity of LAMP, PCR and in vitro culture methods for the detection of Theileria equi and Babesia caballi was evaluated using tenfold serially diluted culture parasites. On day 1 post-culture, both T. equi and B. caballi parasites could only be observed at 1% parasite dilution from the in vitro culture method, whereas LAMP could detect up to 1 × 10⁻³% of both T. equi and B. caballi parasite dilutions, whilst PCR could detect 1 × 10⁻³% T. equi and 1 × 10⁻¹% B. caballi parasite dilutions. On day 7 post-culture, the detection limit for T. equi and B. caballi in the in vitro culture increased up to 1 × 10⁻⁶%, whereas LAMP detection limit increased to 1 × 10⁻¹⁰% for both parasites, whilst the PCR detection limit increased to 1 × 10⁻¹⁰% and 1 × 10⁻⁶% for T. equi and B. caballi, respectively. Furthermore, LAMP and PCR amplified the T. equi DNA extracted from the organs of an experimentally infected horse. This study further validates LAMP as an alternative molecular diagnostic tool, which can be used in the diagnosis of early infections of equine piroplasmosis and together with PCR can also be used as supplementary methods during post-mortems.     

Echinococcus granulosus: membrane permeability of secondary hydatid cysts to albendazole sulfoxide

The objectives of the present study were, first, to establish a methodology for evaluation of the permeability in vitro of hydatid cysts to different drugs and, second, to compare the permeability to albendazole sulfoxide of cysts from untreated animals, cysts from animals treated with 50 mg/kg netobimin for 5 days, and cysts from animals treated with 50 mg/kg netobimin plus 1.1 mg/kg fenbendazole for 5 days. The drug flow follows the Fick law, i.e., the uptake occurs by simple diffusion. We calculated the permeability constant of the cyst membrane by taking into account the disappearance velocity constant, the cyst area, and the incubation solution volume. The permeability value obtained for albendazole sulfoxide was 8.06 ± 2.30 × 10⁻⁶ cm s⁻¹ in cysts from untreated animals, 5.56 ± 2.53 × 10⁻⁶ cm s⁻¹ in cysts from animals treated with netobimin, and 7.05 ± 3.04 × 10⁻⁶ cm s⁻¹ in cysts from animals treated with netobimin + fenbendazole. These permeability values show significant differences (P < 0.05). Received: 20 September 1997 / Accepted: 15 October 1997     

Mutation of the herpes simplex virus 1 KOS UL45 gene reveals dose dependent effects on central nervous system growth

Summary.  The herpes simplex virus type 1 (HSV-1) UL45 gene encodes an 18 kDa virion envelope protein whose function remains unknown. Previous studies using a UL45 null mutant, UL45Δ, demonstrated that deletion of the UL45 gene altered plaque size in Vero and HeLa cells, but was not essential for replication in these cell types. The goal of this study was to determine if mutation of the UL45 gene influenced virus growth in the CNS. Two UL45 mutants, as well as a repaired revertant virus, were constructed and tested for their ability to cause encephalitis and replicate in the CNS. The UL45 mutants were not lethal when 1 × 10³ pfu were injected intracerebrally into Balb/c mice. In contrast, at inocula greater than 1 × 10³, the UL45 mutants were lethal. In vivo growth curves derived from mice inoculated intracerebrally with 1 × 10³ pfu of virus revealed that the mutants grew poorly compared to wild type or revertant viruses. These results suggest that the 18 kDa UL45 gene product is required for efficient growth in the central nervous system at low doses. We propose that the UL45 gene may play an important role under the conditions of a naturally acquired infection. Received June 23, 2001 Accepted November 1, 2001     

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.     

Measurement and prediction of peak shivering intensity in humans

Prediction equations of shivering metabolism are critical to the development of models of thermoregulation during cold exposure. Although the intensity of maximal shivering has not yet been predicted, a peak shivering metabolic rate (Shiv_peak) of five times the resting metabolic rate has been reported. A group of 15 subjects (including 4 women) [mean age 24.7 (SD 6) years, mean body mass 72.1 (SD 12) kg, mean height 1.76 (SD 0.1) m, mean body fat 22.3 (SD 7)% and mean maximal oxygen uptake (V˙O_2max) 53.2 (SD 9) ml O₂ · kg⁻¹ · min⁻¹] participated in the present study to measure and predict Shiv_peak. The subjects were initially immersed in water at 8°C for up to 70 min. Water temperature was then gradually increased at 0.8 °C · min⁻¹ to a value of 20 °C, which it was expected would increase shivering heat production based on the knowledge that peripheral cold receptors fire maximally at approximately this temperature. This, in combination with the relatively low core temperature at the time this water temperature was reached, was hypothesized would stimulate Shiv_peak. Prior to warming the water from 8 to 20 °C, the oxygen consumption was 15.1 (SD 5.5) ml · kg⁻¹ · min⁻¹ at core temperatures of approximately 35 °C. After the water temperature had risen to 20 °C, the observed Shiv_peak was 22.1 (SD 4.2) ml O₂ · kg⁻¹ · min⁻¹ at core and mean skin temperatures of 35.2 (SD 0.9) and 22.1 (SD 2.2) °C, respectively. The Shiv_peak corresponded to 4.9 (SD 0.8) times the resting metabolism and 41.7 (SD 5.1)% of V˙O_2max. The best fit equation predicting Shiv_peak was Shiv_peak (ml O₂ · kg⁻¹ · min⁻¹)=30.5 + 0.348 ×V˙O_2max (ml O₂ · kg⁻¹ · min⁻¹) − 0.909 × body mass index (kg · m⁻²) − 0.233 × age (years); (P=0.0001; r ²=0.872). Accepted: 7 September 2000     

Comparative Cell Shape and Diffusional Water Permeability of Red Blood Cells from Indian Elephant ( Elephas maximus ) and Man ( Homo sapiens )

Red blood cells (RBC) from an Indian elephant (Elephas maximus) were studied by light microscopy (LM), scanning electron microscopy (SEM) and a new nuclear magnetic resonance (NMR) ‘imaging’ method based on the translational diffusion of water, NMR q-space analysis. Also, the transmembrane diffusional permeability, P _d of water in RBC was measured by using a Mn²⁺-doping NMR technique, taking human RBC as a reference. The main diameter of the elephant RBC was measured as 9.3 ± 0.7 μm by LM, 9.3 ± 0.7 μm by ‘shrinkage-corrected’ SEM, and 9.3 ± 0.4 μm by q-space anlaysis. The value is ∼1.4 μm larger than that for the human RBC. The values of P _d were, in the case of elephant RBC, 3.2 × 10⁻³ cm/s at 25 °C, 3.9 × 10⁻³ cm/s at 30 °C, 5.2 × 10⁻³ cm/s at 37 °C and 6.5 × 10⁻³ cm/s at 42 °C; all values were significantly lower than the corresponding values of P _d for human RBC, namely 4.3 × 10⁻³ cm/s at 25 °C, 5.2 × 10⁻³ cm/s at 30 °C, 6.1 × 10⁻³ cm/s at 37 °C, 7.8 × 10⁻³ cm/s at 42°C. The maximal inhibition of P _d (56%) was reached in 30 min at 37 °C with 2 mm p-chloromercuribenzene sulphonate (PCMBS) for both species of RBC. The basal permeability to water at 37 °C was estimated to be 2.3 × 10⁻³ cm/s for elephant and 2.6 × 10⁻³ cm/s for human RBC. The values of the activation energy for water permeability (E _a,d ) was significantly higher for elephant RBC (31.9 kJ/mol) than for human RBC (25.9 kJ/mol). This indicated that features other than the number of transporters per cell are likely to be important in defining the differences in water permeability in the RBC from the two species.     

Comparative Cell Shape and Diffusional Water Permeability of Red Blood Cells from Indian Elephant (Elephas maximus) and Man (Homo sapiens)

Red blood cells (RBC) from an Indian elephant (Elephas maximus) were studied by light microscopy (LM), scanning electron microscopy (SEM) and a new nuclear magnetic resonance (NMR) ‘imaging’ method based on the translational diffusion of water, NMR q-space analysis. Also, the transmembrane diffusional permeability, P _d of water in RBC was measured by using a Mn²⁺-doping NMR technique, taking human RBC as a reference. The main diameter of the elephant RBC was measured as 9.3 ± 0.7 μm by LM, 9.3 ± 0.7 μm by ‘shrinkage-corrected’ SEM, and 9.3 ± 0.4 μm by q-space anlaysis. The value is ∼1.4 μm larger than that for the human RBC. The values of P _d were, in the case of elephant RBC, 3.2 × 10⁻³ cm/s at 25 °C, 3.9 × 10⁻³ cm/s at 30 °C, 5.2 × 10⁻³ cm/s at 37 °C and 6.5 × 10⁻³ cm/s at 42 °C; all values were significantly lower than the corresponding values of P _d for human RBC, namely 4.3 × 10⁻³ cm/s at 25 °C, 5.2 × 10⁻³ cm/s at 30 °C, 6.1 × 10⁻³ cm/s at 37 °C, 7.8 × 10⁻³ cm/s at 42°C. The maximal inhibition of P _d (56%) was reached in 30 min at 37 °C with 2 mm p-chloromercuribenzene sulphonate (PCMBS) for both species of RBC. The basal permeability to water at 37 °C was estimated to be 2.3 × 10⁻³ cm/s for elephant and 2.6 × 10⁻³ cm/s for human RBC. The values of the activation energy for water permeability (E _a,d ) was significantly higher for elephant RBC (31.9 kJ/mol) than for human RBC (25.9 kJ/mol). This indicated that features other than the number of transporters per cell are likely to be important in defining the differences in water permeability in the RBC from the two species.     

Total energy expenditure of elite synchronized swimmers measured by the doubly labeled water method

To determine the daily energy requirement of elite synchronized swimmers during moderate-intensity training, the average daily energy expenditure measured by the doubly labeled water method, was calculated for nine female Japanese national team synchronized swimmers [four senior; mean (SD) 22.5 (1.0) years old, 52.2 (3.6) kg, and five junior; 17.6 (1.1) years old, 52.8 (2.3) kg]. Their total energy expenditure (TEE) was 11.5 (2.8) MJ · day⁻¹ [2738 (672) kcal · day⁻¹]. When compared with estimated energy requirements derived from “Recommended Dietary Allowances for the Japanese”, 12.1 (0.6) MJ · day⁻¹ [2897 (139) kcal · day⁻¹], there was no difference between mean actual and estimated energy requirements. However, there were considerable differences observed on an individual basis. Their energy intake, estimated from 7- day self-reported dietary records, was 8.9 (1.7) MJ · day⁻¹ [2128 (395) kcal · day⁻¹], which was significantly lower than their TEE (P < 0.05). Resting energy expenditure (REE), as determined by indirect calorimetry, was 5.2 (0.3) MJ · day⁻¹ [1247 (75) kcal · day⁻¹]. Their physical activity level (TEE/REE) was 2.18 (0.43). These results demonstrate that the TEE values of elite female synchronized swimmers are not dissimilar to those reported for athletes participating in other sports, especially competitive swimmers during moderate-intensity training. Accepted: 26 May 2000     

Genome-wide Association with C-Reactive Protein Levels in CLHNS: Evidence for the CRP and HNF1A Loci and their Interaction with Exposure to a Pathogenic Environment

Recent genome-wide association studies have related several genetic loci, including C-reactive protein (CRP), hepatocyte nuclear factor 1 homeobox (HNF1A), and genetic variations in the leptin receptor (LEPR), to circulating CRP levels in populations of European ancestry. The genetic effects in other populations and across varying levels of exposure to a pathogenic environment, an important environmental factor associated with CRP, remain to be determined. We tested 2,073,674 single-nucleotide polymorphisms (SNPs) for association with plasma CRP (limited to ≤10 mg/L) in 1,709 unrelated Filipino women from the Cebu Longitudinal Health and Nutrition Survey. The strongest evidence of association was observed with variants at CRP (rs876537, P = 1.4 × 10⁻⁹) and HNF1A (rs7305618, P = 1.0 × 10⁻⁸). Among other previously reported CRP-associated loci, the apolipoprotein E ε4 haplotype was associated with decreased CRP level (P = 7.1 × 10⁻⁴), and modest association was observed with LEPR (rs1892534, P = 0.076), with direction of effects consistent with previous studies. The strongest signal at a locus not previously reported mapped to a gene desert region on chromosome 6q16.1 (rs1408282, P = 2.9 × 10⁻⁶). Finally, we observed nominal evidence of interaction with exposure to a pathogenic environment for top main effect SNPs at HNF1A (rs7305618, P = 0.031), LEPR (rs1892535, P = 0.030) and 6q16.1 (rs1408282, P = 0.046). Our findings demonstrate convincing evidence that genetic variants in CRP and HNF1A contribute to plasma CRP in Filipino women and provide the first evidence that exposure to a pathogenic environment may modify the genetic influence at the HNF1A, LEPR, and 6q16.1 loci on plasma CRP level.     

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     

Muscle glycogen resynthesis rate in humans after supplementation of drinks containing carbohydrates with low and high molecular masses

The rate of muscle glycogen synthesis during 2 and 4 h of recovery after depletion by exercise was studied using two energy equivalent carbohydrate drinks, one containing a polyglucoside with a mean molecular mass of 500 000–700 000 (C drink), and one containing monomers and oligomers of glucose with a mean molecular mass of approximately 500 (G drink). The osmolality was 84 and 350 mosmol · l⁻¹, respectively. A group of 13 healthy well-trained men ingested the drinks after glycogen depleting exercise, one drink at each test occasion. The total amount of carbohydrates consumed was 300 g (4.2 g · kg⁻¹) body mass given as 75 g in 500 ml water immediately after exercise and again 30, 60 ad 90-min post exercise. Blood glucose and insulin concentrations were recorded at rest and every 30 min throughout the 4-h recovery period. Muscle biopsies were obtained at the end of exercise and after 2 and 4 h of recovery. Mean muscle glycogen contents after exercise were 52.9 (SD 27.4) mmol glycosyl units · kg⁻¹ (dry mass) in the C group and 58.3 (SD 35.4) mmol glycosyl units · kg⁻¹ (dry mass) in the G group. Mean glycogen synthesis rate was significantly higher during the initial 2 h for the C drink compared to the G drink: 50.2 (SD 13.7) mmol · kg⁻¹ (dry mass) · h⁻¹ in the C group and 29.9 (SD 12.5) mmol · kg⁻¹ (dry mass) · h⁻¹ in the G group. During the last 2 h the mean synthesis rate was 18.8 (SD 33.3) and 23.3 (SD 22.4) mmol · kg⁻¹ (dry mass) · h⁻¹ in the C and G group, respectively (n.s.). Mean blood glucose and insulin concentrations did not differ between the two drinks. Our data indicted that the osmolality of the carbohydrate drink may influence the rate of resynthesis of glycogen in muscle after its depletion by exercise. Accepted: 9 September 1999     

Effect of creatine supplementation on metabolism and performance in humans during intermittent sprint cycling

This double blind study investigated the effect of oral creatine supplementation (CrS) on 4 × 20 s of maximal sprinting on an air-braked cycle ergometer. Each sprint was separated by 20 s of recovery. A group of 16 triathletes [mean age 26.6 (SD 5.1) years. mean body mass 77.0 (SD 5.8) kg, mean body fat 12.9 (SD 4.6)%, maximal oxygen uptake 4.86 (SD 0.7) l · min⁻¹] performed an initial 4 × 20 s trial after a muscle biopsy sample had been taken at rest. The subjects were then matched on their total intramuscular creatine content (TCr) before being randomly assigned to groups to take by mouth either a creatine supplement (CRE) or a placebo (CON) before a second 4 × 20 s trial. A muscle biopsy sample was also taken immediately before this second trial. The CrS of 100 g comprised 4 × 5 g for 5 days. The initial mean TCr were 112.5 (SD 8.7) and 112.5 (SD 10.7) mmol · kg⁻¹ dry mass for CRE and CON, respectively. After creatine loading and placebo ingestion respectively, CRE [128.7 (SD 11.8) mmol · kg⁻¹ dry mass] had a greater (P=0.01) TCr than CON [112.0 (SD 10.0) mmol · kg⁻¹ dry mass]. While the increase in free creatine for CRE was statistically significant (P=0.034), this was not so for the changes in phosphocreatine content [trial 1: 75.7 (SD 6.9), trial 2: 84.7 (SD 11.0) mmol · kg⁻¹ dry mass, P=0.091]. There were no significant differences between CRE and CON for citrate synthase activity (P=0.163). There was a tendency towards improved performance in terms of 1 s peak power (in watts P=0.07; in watts per kilogram P=0.05), 5 s peak power (in watts P=0.08) and fatigue index (P=0.08) after CrS for sprint 1 of the second trial. However, there was no improvement for mean power (in watts P=0.15; in watts per kilogram P=0.1) in sprint 1 or for any performance values in subsequent sprints. Our results suggest that, while CrS elevates the intramuscular stores of free creatine, this does not have an ergogenic effect on 4 × 20 s all-out cycle sprints with intervening 20-s rest periods. Accepted: 2 October 2000     

Specificity of treadmill and cycle ergometer tests in triathletes, runners and cyclists

The objective of this study was to evaluate the viability of using a single test in which cardiorespiratory variables are measured, to establish training guidelines in running and/or cycling training activities. Six triathletes (two females and four males), six runners (two females and four males) and six males cyclists, all with 5.5 years of serious training and still involved in racing, were tested on a treadmill and cycle ergometer. Cardiorespiratory variables [e.g., heart rate (HR), minute ventilation, carbon dioxide output (V˙CO₂)] were calculated relative to fixed percentages of maximal oxygen uptake (V˙O_2max; from 50 to 100%). The entire group of subjects had significantly (P < 0.05) higher values of V˙O_2max on the treadmill compared with the cycle ergometer [mean (SEM) 4.7 (0.8) and 4.4 (0.9) l · min⁻¹, respectively], and differences between tests averaged 10.5% for runners, 6.1% for triathletes and 2.8% for cyclists. A three-way analysis of variance using a 3 × 2 × 6 design (groups × tests × intensities) demonstrated that all factors yielded highly significant F-ratios (P < 0.05) for all variables between tests, even though differences in HR were only 4 beats · min⁻¹. When HR was plotted against a fixed percentage of V˙O_2max, a high correlation was found between tests. These results demonstrate that for triathletes, cyclists and runners, the relationship between HR and percentage of V˙O_2max, obtained in either a treadmill or a cycle ergometer test, may be used independently of absolute V˙O_2max to obtain reference HR values that can be used to monitor their running and/or cycling training bouts. Received: 3 November 1998 / Accepted: 29 July 1999     

Tamoxifen treatment of myocardial infarcted female rats exacerbates scar formation

Hormonal replacement therapy in postmenopausal women was associated with an increased incidence of nonfatal myocardial infarction. Selective estrogen receptor modulators were considered an alternative pharmacological approach. However, selective estrogen receptor modulators acting via estrogen receptor-dependent and receptor-independent mechanisms may negatively influence cardiac remodeling. The present study tested the hypothesis that tamoxifen (TAM) treatment after coronary artery ligation compromised scar formation. TAM administration (10 mg kg⁻¹ day⁻¹ for 3 weeks) to postmyocardial infarcted (MI) female adult rats significantly increased scar surface area (TAM+MI = 0.67 ± 0.08 vs MI = 0.45 ± 0.06 cm²) and weight (TAM+MI = 0.071 ± 0.007 vs MI = 0.050 ± 0.006 grams). In the infarct region, a significant decrease (p < 0.05) of small calibre vessels (lumen diameter <50 μm) was observed in TAM treated post-MI rats (4.5 ± 0.8 vessels/mm²), as compared to untreated MI rats (7 ± 0.7 vessels/mm²). Consistent with the latter finding, 4-OH TAM caused a dose-dependent suppression of vascular endothelial growth factor (VEGF)-stimulated (10⁻⁹ mol/l) capillarity-like tubule formation by rat aortic endothelial cells in vitro via an estrogen receptor-independent mechanism. These data have demonstrated that TAM treatment of post-MI female rats exacerbated scar formation and may have occurred at least in part via the attenuation of new vessel formation in the infarct region.     

The oxygen consumption with unloaded walking and load carriage using two different backpack designs

The purpose of this study was to assess the energy expenditure associated with load carriage using both a traditional rucksack and a new rucksack design, the AARN rucksack, which incorporates front balance pockets. Nine volunteers walked at 3 km h⁻¹ at various uphill and downhill gradients on a treadmill without a load and carrying a load of 25.6 kg in each of the rucksacks. The oxygen consumption associated with both of the loading conditions was significantly (P < 0.001) higher than that associated with unloaded walking at all downhill gradients tested, although there was no significant difference between the two loading conditions. During the uphill gradients the oxygen consumption associated with the AARN pack was significantly (P < 0.05) lower than that associated with the traditional pack at the 0%, 5%, 10% and 20% gradients. The mean (%) differences at these gradients, expressed in ml · kg⁻¹ · min⁻¹ were 1.18 (9%), 1.45 (8%), 1.76 (8%) and 1.88 (6%), respectively. On average for the whole protocol, the oxygen consumption associated with the AARN rucksack was 5% lower than that associated with the traditional rucksack [mean (SD) 17.28 (7.46) ml · kg⁻¹ · min⁻¹ for the AARN pack and 18.20 (7.84) ml · kg⁻¹ · min⁻¹ for the traditional pack]. The findings of the present study suggest that a load carriage system that allows the load to be distributed between the back and font of the trunk is more appropriate for carrying relatively heavy loads than a system that loads the back only. Accepted: 22 November 1999     

Endothelial function in aorta segments of apolipoprotein E-deficient mice before development of atherosclerotic lesions

Acetylcholine (ACh)-induced relaxation declines in apolipoprotein E-deficient (apoE^−/−) mouse aortas, but only after atherosclerotic plaque formation. This study investigated intracellular calcium concentrations [Ca²⁺]_i and changes in phenylephrine-induced contractions as index of baseline nitric oxide (NO) bioavailability before plaque development. Isometric contractions of thoracic aorta rings of young (4 months) apoE^−/− and C57BL/6J (WT) mice were evoked by phenylephrine (3 × 10⁻⁹–3 × 10⁻⁵ M) in the presence and absence of endothelial cells (ECs) or NO synthase (NOS) inhibitors. [Ca²⁺]_i (Fura-2 AM) and endothelium-dependent relaxation were measured at baseline and after ACh stimulation. Segments of apoE^−/− mice were significantly more sensitive and developed more tension than WT segments in response to phenylephrine. The differences disappeared after NOS inhibition or EC removal or upon increasing [Ca²⁺]_i in apoE^−/− strips with 10⁻⁶ M cyclopiazonic acid or 10⁻⁷ M Ca²⁺-ionophore A23187. Expression of endothelial NOS (eNOS) mRNA was similar in apoE^−/− and WT aorta segments. Basal [Ca²⁺]_i was significantly lower in apoE^−/− than in WT strips. Relaxation by ACh (3 × 10⁻⁹–10⁻⁵ M) was time- and dose-dependently related to [Ca²⁺]_i, but neither ACh-induced relaxation nor Ca²⁺ mobilization were diminished in apoE^−/− strips. In conclusion, basal, but not ACh-induced NO bioavailability, was compromised in lesion-free aorta of apoE^−/− mice. Decreased basal NO bioavailability was not related to lower eNOS expression, but most likely related to lower basal [Ca²⁺]_i. These findings further point to important differences between basal and stimulated eNOS activity.     

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