Serum osmolality measured in fingertip capillary blood is comparable to serum osmolality measured in venous blood.

Blood osmolality is considered the gold standard hydration assessment, but has limited application for technical and invasive reasons. Paired antecubital-venous blood and fingertip-capillary blood were collected pre- and 30 min post-drinking 600 mL water in 55 male/female participants. No bias (0.2 mOsmo/kg, limits of agreement = -2.5 to 2.8 mOsmo/kg) was found between sampling methods, with high linear correlation (Spearman's r = 0.95, P < 0.001). Capillary blood sampling offers an accurate less-invasive method for determining serum osmolality than venous blood sampling.

Ischaemic preconditioning improves upper-body endurance performance without altering V̇O2 kinetics.

PURPOSE: Whilst pre-exercise ischaemic preconditioning (IPC) can improve lower-body exercise performance, its impact on upper-limb performance has received little attention. This study examines the influence of IPC on upper-body exercise performance and oxygen uptake (V̇O2) kinetics. METHODS: Eleven recreationally-active males (24 ± 2 years) completed an arm-crank graded exercise test to exhaustion to determine the power outputs at the ventilatory thresholds (VT1 and VT2) and V̇O2peak (40.0 ± 7.4 ml·kg-1·min-1). Four main trials were conducted, two following IPC (4 × 5-min, 220 mmHg contralateral upper-limb occlusion), the other two following SHAM (4 × 5-min, 20 mmHg). The first two trials consisted of a 15-minute constant work rate and the last two time-to-exhaustion (TTE) arm-crank tests at the power equivalents of 95% VT1 (LOW) and VT2 (HIGH), respectively. Pulmonary V̇O2 kinetics, heart rate, blood-lactate concentration, and rating of perceived exertion were recorded throughout exercise. RESULTS: TTE during HIGH was longer following IPC than SHAM (459 ± 115 vs 395 ± 102 s, p = .004). Mean response time and change in V̇O2 between 2-min and end exercise (ΔV̇O2) were not different between IPC and SHAM for arm-cranking at both LOW (80.3 ± 19.0 vs 90.3 ± 23.5 s [p = .06], 457 ± 184 vs 443 ± 245 ml [p = .83]) and HIGH (96.6 ± 31.2 vs 92.1 ± 24.4 s [p = .65], 617 ± 321 vs 649 ± 230 ml [p = .74]). Heart rate, blood-lactate concentration, and rating of perceived exertion did not differ between conditions (all p ≥ .05). CONCLUSION: TTE was longer following IPC during upper-body exercise despite unchanged V̇O2 kinetics.HighlightsWhilst pre-exercise ischaemic preconditioning can improve lower-body exercise performance and alter V̇O2 kinetics, its impact on upper-limb performance has received little attention.An acute bout of ischaemic preconditioning prior to arm-crank ergometry exercise significantly improved time to exhaustion compared to a sham control condition.V̇O2 kinetics in response to ischaemic preconditioning remained unchanged, suggesting alternative mechanisms may explain performance improvements.

A 7-day high-fat, high-calorie diet induces fibre-specific increases in intramuscular triglyceride and perilipin protein expression in human skeletal muscle.

KEY POINTS: We have recently shown that a high-fat, high-calorie (HFHC) diet decreases whole body glucose clearance without impairing skeletal muscle insulin signalling, in healthy lean individuals. These diets are also known to increase skeletal muscle IMTG stores, but the effect on lipid metabolites leading to skeletal muscle insulin resistance has not been investigated. This study measured the effect of 7 days' HFHC diet on (1) skeletal muscle concentration of lipid metabolites, and (2) potential changes in the perilipin (PLIN) content of the lipid droplets storing intramuscular triglyceride (IMTG). The HFHC diet increased PLIN3 protein expression and redistributed PLIN2 to lipid droplet stores in type I fibres. The HFHC diet increased IMTG content in type I fibres, while lipid metabolite concentrations remained the same. The data suggest that the increases in IMTG stores assists in reducing the accumulation of lipid metabolites known to contribute to skeletal muscle insulin resistance. ABSTRACT: A high-fat, high-calorie (HFHC) diet reduces whole body glucose clearance without impairing skeletal muscle insulin signalling in healthy lean individuals. HFHC diets also increase skeletal muscle lipid stores. However, unlike certain lipid metabolites, intramuscular triglyceride (IMTG) stored within lipid droplets (LDs) does not directly contribute to skeletal muscle insulin resistance. Increased expression of perilipin (PLIN) proteins and colocalisation to LDs has been shown to assist in IMTG storage. We aimed to test the hypothesis that 7 days on a HFHC diet increases IMTG content while minimising accumulation of lipid metabolites known to disrupt skeletal muscle insulin signalling in sedentary and obese individuals. We also aimed to identify changes in expression and subcellular distribution of proteins involved in IMTG storage. Muscle biopsies were obtained from the m. vastus lateralis of 13 (11 males, 2 females) healthy lean individuals (age: 23 ± 2.5 years; body mass index: 24.5 ± 2.4 kg m-2 ), following an overnight fast, before and after consuming a high-fat (64% energy), high-calorie (+47% kcal) diet for 7 days. After the HFHC diet, IMTG content increased in type I fibres only (+101%; P < 0.001), whereas there was no change in the concentration of either total diacylglycerol (P = 0.123) or total ceramides (P = 0.150). Of the PLINs investigated, only PLIN3 content increased (+50%; P < 0.01) solely in type I fibres. LDs labelled with PLIN2 increased (+80%; P < 0.01), also in type I fibres only. We propose that these adaptations of LDs support IMTG storage and minimise accumulation of lipid metabolites to protect skeletal muscle insulin signalling following 7 days' HFHC diet.

Heterogeneous recruitment of quadriceps muscle portions and fibre types during moderate intensity knee-extensor exercise: effect of thigh occlusion.

The involvement of quadriceps femoris muscle portions and fibre type recruitment was studied during submaximal knee-extensor exercise without and with thigh occlusion (OCC) and compared with responses during intense exercise. Six healthy male subjects performed 90-s of moderate exercise without (MOD; 29+/-4 W) and with thigh OCC, and moderate exercise followed by 90-s of intense exercise (HI; 65+/-8 W). Temperatures were continuously measured in m. vastus lateralis (VL), vastus medialis (VM) and rectus femoris (RF) and successive muscle biopsies were obtained from VL. During MOD, muscle temperature increase (DeltaT(m)) in RF was 0.52+/-0.09 degrees C, which was 57% and 73% higher (P<0.05) than in VL and VM, respectively. During OCC, DeltaT(m) in RF was 0.39+/-0.05 degrees C, which was not different from VM but 54% higher (P<0.05) than in VL. After MOD, muscle CP in slow twitch (ST) and fast twitch (FT) fibres was 81% and 91% of resting levels, respectively, with lower (P<0.05) values after OCC (15% and 22%) and HI (24% and 13%). After MOD, OCC and HI, a total of 48%, 93% and 96% of the ST fibres had CP levels below mean-1 SD, respectively, with corresponding values for FT fibres being 41%, 89% and 100%, respectively. In conclusion, a heterogeneous recruitment of the quadriceps muscle portions and muscle fibres was observed during submaximal knee-extensor exercise, whereas recruitment pattern was homogenous during intense exercise. Thigh OCC caused an altered recruitment of fibres and muscle portions, suggesting a significant afferent response affecting the activation of fibres in the contracting muscles.

Effect of temperature on skeletal muscle energy turnover during dynamic knee-extensor exercise in humans.

The present study examined the effect of elevated temperature on muscle energy turnover during dynamic exercise. Nine male subjects performed 10 min of dynamic knee-extensor exercise at an intensity of 43 W (SD 10) and a frequency of 60 contractions per minute. Exercise was performed under normal (C) and elevated muscle temperature (HT) through passive heating. Thigh oxygen uptake (V(O2)) was determined from measurements of thigh blood flow and femoral arterial-venous differences for oxygen content. Anaerobic energy turnover was estimated from measurements of lactate release as well as muscle lactate accumulation and phosphocreatine utilization based on analysis of muscle biopsies obtained before and after each exercise. At the start of exercise, muscle temperature was 34.5 degrees C (SD 1.7) in C compared with 37.2 degrees C (SD 0.5) during HT (P < 0.05). Thigh V(O2) after 3 min was 0.52 l/min (SD 0.11) in C and 0.63 l/min (SD 0.13) in HT, and at the end of exercise it was 0.60 l/min (SD 0.14) and 0.61 l/min (SD 0.10) in C and HT, respectively (not significant). Total lactate release was the same between the two temperature conditions, as was muscle lactate accumulation and PCr utilization. Total ATP production (aerobic + anaerobic) was the same between each temperature condition [505.0 mmol/kg (SD 107.2) vs. 527.1 mmol/kg (SD 117.6); C and HT, respectively]. In conclusion, within the range of temperatures studied, passively increasing muscle temperature before exercise has no effect on muscle energy turnover during dynamic exercise.

Muscle oxygen uptake and energy turnover during dynamic exercise at different contraction frequencies in humans.

1. It has been established that pulmonary oxygen uptake is greater during cycle exercise in humans at high compared to low contraction frequencies. However, it is unclear whether this is due to more work being performed at the high frequencies and whether the energy turnover of the working muscles is higher. The present study tested the hypothesis that human skeletal muscle oxygen uptake and energy turnover are elevated during exercise at high compared to low contraction frequency when the total power output is the same. 2. Seven subjects performed single-leg dynamic knee-extensor exercise for 10 min at contraction frequencies of 60 and 100 r.p.m. where the total power output (comprising the sum of external and internal power output) was matched between frequencies (54 +/- 5 vs. 56 +/- 5 W; mean +/- S.E.M.). Muscle oxygen uptake was determined from measurements of thigh blood flow and femoral arterial - venous differences for oxygen content (a-v O(2) diff). Anaerobic energy turnover was estimated from measurements of lactate release and muscle lactate accumulation as well as muscle ATP and phosphocreatine (PCr) utilisation based on analysis of muscle biopsies obtained before and after each exercise bout. 3. Whilst a-v O(2) diff was the same between contraction frequencies during exercise, thigh blood flow was higher (P < 0.05) at 100 compared to 60 r.p.m. Thus, muscle V(O2) was higher (P < 0.05) during exercise at 100 r.p.m. Muscle V(O2) increased (P < 0.05) by 0.06 +/- 0.03 (12 %) and 0.09 +/- 0.03 l min(-1) (14 %) from the third minute to the end of exercise at 60 and 100 r.p.m., respectively, but there was no difference between the two frequencies. 4. Muscle PCr decreased by 8.1 +/- 1.7 and 9.1 +/- 2.0 mmol (kg wet wt)(-1), and muscle lactate increased to 6.8 +/- 2.1 and 9.8 +/- 2.5 mmol (kg wet wt)(-1) during exercise at 60 and 100 r.p.m., respectively. The total release of lactate during exercise was 48.7 +/- 8.8 and 64.3 +/- 10.6 mmol at 60 and 100 r.p.m. (not significant, NS). The total anaerobic ATP production was 47 +/- 8 and 61 +/- 12 mmol kg(-1), respectively (NS). 5. Muscle temperature increased (P < 0.05) from 35.8 +/- 0.3 to 38.2 +/- 0.2 degrees C at 60 r.p.m. and from 35.9 +/- 0.3 to 38.4 +/- 0.3 degrees C at 100 r.p.m. Between 1 and 7 min muscle temperature was higher (P < 0.05) at 100 compared to 60 r.p.m. 6. The estimated mean rate of energy turnover during exercise was higher (P < 0.05) at 100 compared to 60 r.p.m. (238 +/- 16 vs. 194 +/- 11 J s(-1)). Thus, mechanical efficiency was lower (P < 0.05) at 100 r.p.m. (24 +/- 2 %) compared to 60 r.p.m. (28 +/- 3 %). Correspondingly, efficiency expressed as work per mol ATP was lower (P < 0.05) at 100 than at 60 r.p.m. (22.5 +/- 2.1 vs. 26.5 +/- 2.5 J (mmol ATP)(-1)). 7. The present study showed that muscle oxygen uptake and energy turnover are elevated during dynamic contractions at a frequency of 100 compared with 60 r.p.m. It was also observed that muscle oxygen uptake increased as exercise progressed in a manner that was not solely related to the increase in muscle temperature and lactate accumulation.

Phosphocreatine and ATP content in human single muscle fibres before and after maximum dynamic exercise.

The recovery of high-energy phosphate levels in single human skeletal muscle fibres following short-term maximal (all-out) exercise was investigated. Three male volunteers exercised maximally for 25 s on an isokinetic cycling ergometer. Muscle biopsy samples from the vastus lateralis were collected at rest, immediately post-exercise and at 1.5 min of recovery. The subjects also performed a second exercise bout 1.5 min after the first, on a separate occasion. Single muscle fibres were dissected, characterized and assigned to one of four groups according to their myosin heavy chain (MyHC) isoform content; namely, type I, IIA, IIAx and IIXa (the latter two groups containing either less or more than 50% IIX MyHC). Fibres were analysed for adenosine 5'-triphosphate (ATP), inosine-5'-monophosphate (IMP), phosphocreatine (PCr) and creatine (Cr) levels. Type I fibres had a lower Cr content than type II fibres (P<0.01). Within type II fibres resting [PCr] increased with increasing MyHC IIX isoform content (r=0.59, P<0.01). Post-exercise [PCr] was very low in all fibre groups (P<0.01 versus rest) while great reductions in ATP were also observed (P<0.01 versus rest), especially in the type II fibre groups. [PCr] at 1.5 min of recovery was still lower compared to rest for all fibre groups (P<0.01) especially in the IIAx and IIXa fibres.

Total power output generated during dynamic knee extensor exercise at different contraction frequencies.

A novel approach has been developed for the quantification of total mechanical power output produced by an isolated, well-defined muscle group during dynamic exercise in humans at different contraction frequencies. The calculation of total power output comprises the external power delivered to the ergometer (i.e., the external power output setting of the ergometer) and the "internal" power generated to overcome inertial and gravitational forces related to movement of the lower limb. Total power output was determined at contraction frequencies of 60 and 100 rpm. At 60 rpm, the internal power was 18+/- 1 W (range: 16-19 W) at external power outputs that ranged between 0 and 50 W. This was less (P<0.05) than the internal power of 33+/-2 W (27-38 W) at 100 rpm at 0-50 W. Moreover, at 100 rpm, internal power was lower (P<0.05) at the higher external power outputs. Pulmonary oxygen uptake was observed to be greater (P<0.05) at 100 than at 60 rpm at comparable total power outputs, suggesting that mechanical efficiency is lower at 100 rpm. Thus a method was developed that allowed accurate determination of the total power output during exercise generated by an isolated muscle group at different contraction frequencies.

Arterial blood pressure and forearm vascular conductance responses to sustained and rhythmic isometric exercise and arterial occlusion in trained rock climbers and untrained sedentary subjects.

Cardiovascular responses to sustained and rhythmic (5 s on, 2 s off) forearm isometric exercise to fatigue at 40% maximal voluntary contraction (MVC) and to a period of arterial occlusion were investigated in elite rock climbers (CLIMB) as a trained population compared to non-climbing sedentary subjects (SED). Blood pressure (BP), monitored continuously by Finapres, and forearm blood flow, by venous occlusion plethysmography, were measured and used to calculate vascular conductance. During sustained exercise, times to fatigue were not different between CLIMB and SED. However, peak increases in systolic (S) BP were significantly lower in CLIMB [25 (13) mmHg; (3.3 (1.7) kPa] than in SED [48 (17) mmHg; (6.4 (2.3) kPa] (P < 0.05), with a similar trend for increases in diastolic (D) BP. Immediately after sustained exercise, forearm conductance was higher in CLIMB than SED (P < 0.05) for up to 2 min. During rhythmic exercise, times to fatigue were two fold longer in CLIMB than SED [853 (76) vs 420 (69) s, P < 0.05]. Increases in SBP were not different between groups except during the last quarter of exercise when they fell in CLIMB. Conductance both during and after rhythmic exercise was higher in CLIMB than in SED. Following a 10-min arterial occlusion, peak vascular conductance was significantly greater in CLIMB than SED [0.597 (0.084) vs 0.431 (0.035) ml x min(-1) x 100 ml(-1) x mmHg(-1); P < 0.05]. The attenuated BP response to sustained isometric exercise could be due in part to enhanced forearm vasodilatory capacity, which also supports greater endurance during rhythmic exercise by permitting greater functional hyperaemia in between contraction phases. Such adaptations would all facilitate the ability of rock climbers to perform their task of making repetitive sustained contractions.

Genetic markers of alcohol abuse.

In this paper, we review the current status of genetic markers for the development of alcohol abuse. Family, twin, half-sibling and adoption studies of alcoholic subjects suggest that the heritability of liability to alcoholism is at least 50%. These findings have fuelled intensive investigation in the fields of neurology, biochemistry, genetics and molecular biology aimed at the identification of markers for the risk of alcoholism. The most promising of these are discussed in detail. Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) polymorphisms, specifically the ADH3*1, ADH2*2, and ALDH2*2 genotypes appear to confer a protective effect against alcoholism, most notably in Oriental subjects. Caucasian alcohol abusers and their first-degree relatives exhibit depressed platelet monoamine oxidase activity, the degree of which is greater in Type II than Type I alcoholics. Electrophysiological characteristics of alcoholics and those at risk for developing alcoholism have also been identified, including the reduced amplitude of the event-related brain potential and, after ethanol ingestion, characteristic EEG alpha-wave activity. Lower platelet adenylate cyclase activity is seen in alcoholics compared to controls, presumably as a result of over-expression of an inhibitory G-protein. Markers related to other signal transduction pathways of the central nervous system including the serotoninergic, muscarinic and dopaminergic systems are also discussed. In this group of markers, the putative association between the inheritance of the AI allele of the D2 dopamine receptor and the susceptibility to alcoholism provides the most dramatic illustration of the challenges presently existing in this field of scientific investigation. Current limitations in the definition, diagnosis and classification of alcoholism, the confounding influences of race and gender on association studies, as well as the statistical approach of linkage studies are discussed as they relate to the endeavor to uncover valid genetic markers for the risk of alcoholism.

Ultrasensitive detection of prostate-specific antigen by a time-resolved immunofluorometric assay and the Immulite immunochemiluminescent third-generation assay: potential applications in prostate and breast cancers.

We report an ultrasensitive time-resolved immunofluorometric assay (TRIFA) for prostate-specific antigen (PSA). The assay is an improvement of our previous report (Clin Chem 1993;39:2108-14) and includes the utilization of two monoclonal antibodies and a one-step incubation period, which greatly reduces analysis time. The new method demonstrates a superior lower analytical limit of detection (< or = 1 ng/L), a wide dynamic range, absence of a hook effect at 10(6) ng/L PSA, and equimolarity for free PSA and PSA-antichymotrypsin complex. Also, we have compared several aspects of our TRIFA with a commercially available third-generation assay (Immulite). An evaluation of breast tumor cytosol extracts from 315 patients shows PSA immunoreactivity > 15 ng/g of total protein in 28% and 23% by TRIFA and Immulite analysis, respectively. Both methods demonstrate a significant association between breast tumor PSA immunoreactivity and progesterone and estrogen receptor positivity (P <0.001). Analysis of serum samples obtained for monitoring of postradical prostatectomy patients reveals significant PSA changes at concentrations undetectable by conventional methods. The significance of these results as well as the potential applications of ultrasensitive PSA assays in breast and prostate cancers are discussed.

Gluconeogenesis in trout (Oncorhynchus mykiss) white muscle: purification and characterization of fructose-1,6-bisphosphatase activity in vitro.

Studies of the enzyme fructose-1,6-bisphosphatase (FBPase) of rainbow trout (Oncorhynchus mykiss) have been undertaken in order to illuminate aspects of skeletal muscle gluconeogenesis in these animals. Maximal activities in crude homogenates of several organs suggest that the liver possesses the greatest FBPase activity on a unit g(-1) tissue basis but that the white muscle, owing to its bulk, contributes substantially to whole body FBPase activity. Studies of fructose-6-phosphate-1-kinase (PFK) and FBPase in crude homogenates of several organs suggests an important role for intracellular pH in regulating the relative carbon flux through the FBPase/PFK locus in vivo. Furthermore, a three-step purification scheme is described for trout white muscle FBPase by which a stable and homogeneous (by SDS PAGE) enzyme preparation (isoelectric point = 7.2; molecular weight = 37.6 kd) was obtained. Kinetic studies of the purified enzyme were undertaken at 20°C under conditions reflective of "rest" and "exercise/recovery" intramuscular pH in vivo. Affinity for substrate (F-1,6-P2) was increased (Km = 6.88 versus 2.44 µmol 1-(-1) as was enzyme activity when pH was lowered from 7.0 to 6.5. Various inhibitor metabolites are identified including F-2,6-P2 (mixed-type inhibitor, Ki = 0.201 µmol 1(-1), pH 7.0) and AMP (non-competitive inhibitor, Ki = 0.438 µmol 1(-1), pH 7.0). Inhibition by F-2,6-P2 was strongly alleviated by a reduction in pH from 7.0 to 6.5 (I50 increased from 0.14 to 0.32 µmol 1(-1)). AMP on the other hand was a more potent inhibitor at pH 6.5 but this inhibition was totally reversed under conditions of citrate, NH4 (+) and AMP typical of muscle during recovery from exercise in vivo. In purified white muscle enzyme preparations, FBPase demonstrated maximal activity at pH 6.5 whereas the optimal pH of PFK was 7.0 or greater. Indeed, it appears from these in vitro data that regulation by metabolite levels as well as pH are required for net FBPase flux in vivo. It is concluded, therefore that trout white muscle FBPase demonstrates the potential to play an important enzymatic role in the control of intramuscular gluconeogenesis in these animals. The results are discussed in relation to present knowledge regarding the metabolic responses of trout white muscle to, and its subsequent recovery from, exhaustive exercise.

Energy metabolism in trout red cells: consequences of adrenergic stimulation in vivo and in vitro.

beta-Adrenergic stimulation of salmonid red cells results in a rapid decrease (within 5 min) in the nucleotide triphosphate:haemoglobin ratio (NTP:Hb), which is thereafter maintained at a constant level, presumably through increased ATP turnover via matched aerobic metabolism and energy-consuming processes. Addition of the beta-adrenergic agonist isoproterenol to rainbow trout red cells in vitro leads to a rise in intracellular pH (pHi), a corresponding decrease in extracellular pH (pHe) and an increase in red cell oxygen consumption (MO2). Moreover, the extent to which red cell pHi is maintained constant in the face of an acute extracellular acidosis in vitro or in vivo is proportional to the adrenergically stimulated increase in red cell MO2. In the absence of oxygen, these red cells remain capable of pH regulation, but cannot maintain NTP:Hb constant. As a result, membrane and metabolic functions become uncoupled in the stimulated deoxygenated cells.

Metabolic-membrane coupling in red blood cells of trout: the effects of anoxia and adrenergic stimulation.

Under oxygenated conditions, in vitro, the highly aerobic red cells of the rainbow trout (Salmo gairdneri) exhibit tight coupling between energy (i.e. nucleotide triphosphate, NTP)-consuming and NTP-producing metabolic activity, as shown by strict maintenance of red cell NTP:haemoglobin ratios. This coupling is maintained following adrenergic stimulation of oxygenated red cells when the increased NTP demands of ion transporting systems are met by enhanced energy production via aerobic metabolism. In unstimulated anoxic red cells, membrane-metabolic coupling is preserved via the arrest of NTP-consuming processes. Adrenergic stimulation of anoxic red cells, however, leads to a functional uncoupling of membrane metabolism with the result that NTP levels decline rapidly. At this time, cellular [NTP] is negatively correlated with [Na+]i and [Cl-]i and positively correlated with [K+]i. This, in addition to the fact that the pH of the intracellular compartment is also highly dependent on cellular NTP levels, provides evidence for the integration of energy and membrane metabolisms.

Metabolic energy production during adrenergic pH regulation in red cells of the Atlantic salmon, Salmo salar.

Whole blood from Atlantic salmon was incubated anaerobically at 10 degrees C so as to measure the metabolic activity of the nucleated erythrocytes. An acute extracellular acidosis was produced by adding either an acid solution (sham) or an acid solution with adrenaline (final concentration, 5 x 10(-4) M). The extracellular acidosis produced by the sham solution was transferred to the erythrocytes, whereas with adrenaline, intracellular pH actually increased in the face of a plasma acidosis. Indeed, the extracellular acidosis in the adrenaline-treated blood was significantly higher than that of the sham as a result of net H+ excretion from the erythrocyte. This pH response of the erythrocyte was accompanied by a proportional increase in the O2 consumption of the blood, with no change in lactate production. In comparison to sham-treated cells, the content of erythrocytic nucleotide triphosphates initially decreased upon addition of adrenaline but was thereafter maintained at a constant NTP/Hb ratio presumably due to an increased ATP turnover. In conclusion, it appears that the aerobic rather than anaerobic metabolism of erythrocytes is accelerated upon addition of adrenaline to blood, and that this increased metabolism is involved in fueling the membrane transport processes involved in adrenergic pH regulation of salmonid red cells.

Absolute bioavailability study in the dog with Visacor, a new cardioselective beta blocking drug with intrinsic sympathomimetic activity.

The cardioselective beta-adrenergic blocking drug Visacor (ICI 141,292) was dosed to six beagle dogs in a randomized cross-over manner. Five formulations were examined i.e. a 15 mg/kg intravenous solution, a 50 mg/kg oral solution, and 50, 100 and 200 mg/kg oral powder formulations. Whole blood and urine samples were collected at various times after each dose and analysed for parent drug concentration by a high pressure liquid chromatography procedure. The urine samples were also analysed for parent drug content after hydrolysis with beta-glucuronidase. The normalised intravenous blood levels of ICI 141,292 were found to decay tri-exponentially with a final phase elimination half-life of about 10 h. The computer fitted data showed the drug to possess a high volume of distribution for both the central compartment (54% body weight) and whole body (1384% body weight) indicating the possibility of a high degree of metabolism. The drug clearance following i.v. administration was 196 ml/min and the urinary recovery rate of parent drug was 24% (unhydrolysed) and 40% following hydrolysis with beta-glucuronidase. Following oral dosing at 50 mg/kg (as both powder (C) and solution (B], 100 (D) and 200 (E) mg/kg (as powder) the systemic blood profiles were found to increase with dose. The mean peak blood level attained was 6 +/- 1, 5 +/- 1, 8 +/- 1 and 14 +/- 1 micrograms/ml for formulations, B, C, D and E respectively. The systemic bioavailability of ICI 141,292 was only about 40%. The areas under the curves increased linearly with dose and the elimination phase half-life was unchanged with dose. The calculated half-life (7 h) was apparently shorter after oral administration than after intravenous administration (10 h) but this is probably an artefact dependent on the limit of detection of the assay procedure. At 50 mg/kg there were no significant differences in blood profiles or in the urinary excretion of drug between the solution and powder formulations. However the overall systemic bioavailability was marginally higher with the powder. These observations are consistent for a drug which is cleared by both renal and hepatic elimination processes, which undergoes "first-pass" metabolism on oral dosing and, over the oral dose range studied, obeys linear pharmacokinetics. The significant increase in recovery of parent drug, after hydrolysis of the urine with beta-glucuronidase, indicates that the ICI 141,292 glucuronide conjugate is present to a significant extent. The results also demonstrate that absorption of parent drug from the gastrointestinal tract may not be complete.

Differentation of rat embryo cells in culture: response following acute maternal exposure to teratogens and non-teratogens.

An in vivo-in vitro test system with high sensitivity to teratogens has been developed and validated. A single acute intra-peritoneal injection of teratogens (18) and non-teratogens (13) was administered to pregnant rats on the 12th day after fertilisation, and uteri were removed after 16 h by laparotomy. 34-36 Embryos somites were selected, and mid-brain (CNS) and fore-limb buds (LB) were dissected free and dispersed as single-cell suspensions in Ham's F12 culture medium. The cells were cultured as micromass cell islands for 5 days, and discrete foci of neuronal cells differentiated in CNS cultures and chondrocytes in LB cultures. After 5 days, differentiation as determined by number of stainable foci of differentiated cells and 3H-GABA incorporation in CNS or 35SO4 incorporation in LB and growth (as determined by total protein) were measured. Both differentiation and growth of CNS and LB cultures were markedly reduced following exposure of the dam to teratogens, whereas no significant effect was observed with non-teratogens. One teratogen (amaranth) and one non-teratogen (nitrilotriacetic acid) were classified as false negative and positive, respectively; the sensitivity of the test (proportion of teratogens correct) was therefore 92% and the specificity (proportion of non-teratogens correct) was 94%. Inhibition of growth and differentiation in the rat embryo cell cultures following maternal exposure forms the basis of a short-term in vitro test for teratogens.

Haematological, coagulation and blood chemistry data in red-bellied tamarins Saguinus labiatus.

Haematology, coagulation and clinical chemistry data are reported for a group of male and female red-bellied tamarins (Saguinus labiatus). The tamarins were juvenile and young adults and were bred in captivity. High mean values for activities of alkaline phosphatase, alanine amino-transferase, aspartate aminotransferase and creatine kinase were noted. The findings are compared with data obtained from other members of the family Callitrichidae.

An application of receptor models to the analysis of data on beta-adrenoceptor blockade.

1. The classical single receptor competitive occupancy model accurately describes the joint action of an agonist (isoprenaline) and a beta-adrenoceptor antagonist (propranolol) or some partial agonists (dichlorisoprenaline, practolol) on the positive chronotropic response in rats which have been depleted of catacholamines. 2. The mathematical form of the model suggests that the dissociation constants of classical competitive partial agonists may be assessed using dose ratios by exactly the same method as that currently used for agonist-antagonist interactions, provided that the log dose-response curves are first suitably normalized. 3. Close agreement between the theoretical mathematical models and the experimental data can be demonstrated by statistical fitting for certain beta-adrenoceptor antagonists (propranolol, dichlorisoprenaline, practolol). 4. The model fails to describe the behaviour of other beta-adrenoceptor antagonists (oxprenolol, pindolol). A possible extension of the model to include these drugs is proposed.