Measurement of the positive muon lifetime and determination of the Fermi constant to part-per-million precision.

We report a measurement of the positive muon lifetime to a precision of 1.0 ppm; it is the most precise particle lifetime ever measured. The experiment used a time-structured, low-energy muon beam and a segmented plastic scintillator array to record more than 2×10(12) decays. Two different stopping target configurations were employed in independent data-taking periods. The combined results give τ(µ(+)) (MuLan)=2 196 980.3(2.2) ps, more than 15 times as precise as any previous experiment. The muon lifetime gives the most precise value for the Fermi constant: G(F) (MuLan)=1.166 378 8(7)×10(-5) GeV(-2) (0.6 ppm). It is also used to extract the µ(-)p singlet capture rate, which determines the proton's weak induced pseudoscalar coupling g(P).

Improved measurement of the positive-muon lifetime and determination of the Fermi constant.

The mean life of the positive muon has been measured to a precision of 11 ppm using a low-energy, pulsed muon beam stopped in a ferromagnetic target, which was surrounded by a scintillator detector array. The result, tau(micro)=2.197 013(24) micros, is in excellent agreement with the previous world average. The new world average tau(micro)=2.197 019(21) micros determines the Fermi constant G(F)=1.166 371(6)x10(-5) GeV-2 (5 ppm). Additionally, the precision measurement of the positive-muon lifetime is needed to determine the nucleon pseudoscalar coupling g(P).

Beyond U(crit): matching swimming performance tests to the physiological ecology of the animal, including a new fish 'drag strip'.

Locomotor performance of animals is of considerable interest from management, physiological, ecological and evolutionary perspectives. Yet, despite the extensive commercial exploitation of fishes and interest in the health of various fish stocks, the relationships between performance capacity, natural selection, ecology and physiology are poorly known for fishes. One reason may be the technical challenges faced when trying to measure various locomotor capacities in aquatic species, but we will argue that the slow pace of developing new species-appropriate swim tests is also hindering progress. A technique developed for anadromous salmonids (the U(crit) procedure) has dominated the fish exercise physiology field and, while accounting for major advances in the field, has often been used arbitrarily. Here we propose criteria swimming tests should adhere to and report on several attempts to match swimming tests to the physiological ecology of the animal. Sprint performance measured with a laser diode/photocell timed 'drag strip' is a new method employing new technology and is reported on in some detail. A second new test involves accelerating water past the fish at a constant rate in a traditional swim tunnel/respirometer. These two performance tests were designed to better understand the biology of a bentho-pelagic marine fish, the Atlantic cod (Gadus morhua). Finally, we report on a modified incremental velocity test that was developed to better understand the biology of the blacknose dace (Rhinichthys atratulus), a Nearctic, lotic cyprinid.

Caudal differential pressure as a predictor of swimming speed of cod (Gadus morhua).

We report the results of an experiment designed to investigate the feasibility of using differential pressure to estimate the swimming speed and metabolic rate of Atlantic cod (Gadus morhua). Seven cod were fitted with a miniature differential pressure sensor mounted on one side of the caudal peduncle immediately anterior to the base of the caudal fin rays. Relationships between differential pressure, tailbeat frequency, tailbeat amplitude, swimming speed and rate of oxygen consumption ((O(2))) were determined as a function of the swimming speed of cod swimming at 5 degrees C in a recirculating 'Brett-style' respirometer. Tailbeat differential pressure, tailbeat amplitude and tailbeat frequency were highly correlated with swimming speed. The average or integrated pressure ranged from 0 to 150 Pa for speeds up to 0.8 m s(-1) (1.1 L s(-1), where L is total body length), while the 'pressure difference' (maximum minus minimum pressure) ranged from 0 to 900 Pa. Small changes in swimming speed of less than 0.05 m s(-1) were readily detected as differences in tailbeat pressure. Burst swimming in the respirometer resulted in huge pressure 'bursts' of up to 5000 Pa 'pressure difference'. The rate of oxygen consumption increased exponentially and was highly correlated with swimming speed (r(2)=0.77). The rate of oxygen consumption was also correlated with tailbeat integrated pressure (r(2)=0.68) and with differential pressure (r(2)=0.43); regression correlations were always greater for individuals than for combined data from all cod. The results detailed in this study indicate that an ultrasonic differential pressure transmitter would enable accurate estimates of the swimming speed, rates of oxygen consumption and activity patterns of free-ranging fish in nature.

An overview of the pharmacokinetics of cilomilast (Ariflo), a new, orally active phosphodiesterase 4 inhibitor, in healthy young and elderly volunteers.

The oral pharmacokinetics of cilomilast (Ariflo) were investigated in five separate studies in healthy volunteers. Cilomilast was rapidly absorbed, and pharmacokinetics were dose proportional after single and repeat dosing. The elimination half-life was 7 to 8 hours; accordingly, steady state was reached on the 3rd day of dosing. The degree of accumulation following repeat twice-daily dosing was predictable from the data following a single dose. Although systemic exposure (AUC) was, on average, 21% higher in elderly (65-84 years) compared with young subjects, values for Cmax and t(1/2) were similar, and no difference in tolerability was noted. Single and repeat doses of cilomilast up to and including 15 mg (dosed before or taken between meals) were well tolerated. Dosing with food reduced the rate of absorption without affecting total bioavailability. Hence, tolerability was optimal in the fed state; repeat doses up to and including 30 mg twice daily aftermeals were well tolerated following dose titration.

Warfarin pharmacodynamics unaffected by cilomilast.

OBJECTIVE: To demonstrate a lack of effect of steady-state concentrations of cilomilast, a new oral phosphodiesterase 4 inhibitor for the treatment of chronic obstructive pulmonary disease, on warfarin-induced anticoagulation. METHODS: This 28-day, randomized, double-blind, placebo-controlled, parallel-group study involved 36 healthy men. All volunteers received warfarin once daily on days 1 through 24 of the study. After a standard 5-mg loading dose on days 1 and 2, the warfarin dose was titrated between days 3 and 10 to achieve a stable prothrombin time, expressed as international normalized ratio (INR). Volunteers received either cilomilast 15 mg twice daily or placebo on days 18 through 24. The primary end point was the INR on day 24. RESULTS: On day 24, the mean +/- SEM INR in subjects receiving concurrent warfarin and cilomilast was 1.35 +/- 0.05, compared with 1.38 +/- 0.07 in those receiving concurrent warfarin and placebo. The point estimate (90% CI) for the difference in day 24 INR values between cilomilast and placebo (adjusted for baseline) was 0.02 (90% CI-0.13 to 0.17), which translates to an INR ratio of 1.02 (90% CI 0.91 to 0.13). The 90% confidence interval for the ratio of mean INR (cilomilast:placebo) on day 24 was completely contained within the 25% equivalence range, indicating a lack of interaction between warfarin and cilomilast. The adverse event profiles of warfarin/placebo and warfarin/cilomilast were similar and favorable. CONCLUSIONS: The pharmacodynamics of warfarin are unaffected by coadministration of cilomilast at steady-state concentrations in healthy volunteers.

Costs of locomotion and vertic dynamics of cephalopods and fish.

The world's oceans are three-dimensional habitats that support high diversity and biomass. Because the densities of most of the constituents of life are greater than that of seawater, planktonic and pelagic organisms had to evolve a host of mechanisms to occupy the third dimension. Some microscopic organisms survive at the surface by dividing rapidly in vertically well mixed zones, but most organisms, small and large, have antisinking strategies and structures that maintain vertical position and mobility. All of these mechanisms have energetic costs, ranging from the "foregone metabolic benefits" and increased drag of storing high-energy, low-density lipids to direct energy consumption for dynamic lift. Defining the niches in the mesopelagic zone, understanding evolution, and applying such ecological concepts as optimal foraging require good estimates of these costs. The extreme cases above are reasonably well quantified in fishes, but the energetic costs of dynamic physiological mechanisms like swim bladders are not; nor are the costs of maintaining vertical position for the chief invertebrate competitors, the cephalopods. This article evaluates a matrix of buoyancy mechanisms in different circumstances, including vacuum systems and ammonium storage, based on new data on the metabolic cost of creating buoyancy in Sepia officinalis.

The protective effects of hypoxia-induced hypometabolism in the Nautilus.

Specimens of Nautilus pompilius were trapped at depths of 225-300 m off the sunken barrier reef southeast of Port Moresby, Papua New Guinea. Animals transported to the Motupore Island laboratory were acclimated to normal habitat temperatures of 18 degrees C and then cannulated for arterial and venous blood sampling. When animals were forced to undergo a period of progressive hypoxia eventually to encounter ambient partial pressure of oxygen (PO2) levels of approximately 10 mmHg (and corresponding arterial PO2's of approximately 5 mmHg), they responded by lowering their aerobic metabolic rates to 5-10% of those seen in resting normoxic animals. Coincident with this profound metabolic suppression was an overall decrease in activity, with brief periods of jet propulsion punctuating long periods of rest. Below ambient PO2 levels of 30-40 mmHg, ventilatory movements became highly periodic and at the lowest PO2 levels encountered, ventilation occasionally ceased altogether. Cardiac output estimated by the Fick equation decreased during progressive hypoxia by as much as 75 80%, and in the deepest hypometabolic states heart rates slowed to one to two cycles of very low amplitude per minute. By the end of 500 min exposure to ambient PO2 levels of 10 mmHg or less, the anaerobic end products octopine and succinate had increased significantly in adductor muscle and heart, respectively. Increased concentrations of octopine in adductor muscle apparently contributed to a small intracellular acidosis and to the development of a combined respiratory and metabolic acidosis in the extracellular compartment. On the other hand, increases in succinate in heart muscle occurred in the absence of any change in cardiac pHi. Taken together, we estimate that these anaerobic end products would make up less than 2% of the energy deficit arising from the decrease in aerobic metabolism. Thus, metabolic suppression is combined with a massive downregulation of systemic O2 delivery to match metabolic supply to demand.

Simple box model for dense-gas dispersion in a straight sloping channel.

A box model for instantaneous release and subsequent one-dimensional spreading of isothermal dense gases on sloping surfaces is presented. A numerical solution and an approximate analytical solution of the model equations are compared to the experimental data obtained in a sloping heavy-gas channel of the Institute of Fluid Dynamics at ETH-Zürich. The influence of the rear wall of the containment from where the cloud is released is analysed. Different entrainment assumptions, in particular the scaling of the entrainment parameters, are discussed. The numerical values of the entrainment parameters are tuned by computer optimization in order to obtain best agreement of the theoretical results with experimental data.

Choreography of the squid's "nuptial dance".

A mass spawning of squid resembles, at first glance, a chaotic "nuptial dance" (1). But for the first time, we have applied 3-D, radio-linked acoustic positioning (RAP) to this confusing process, and our early results now reveal a choreography that is, in fact, well organized in time and space. Remote tracking with RAP of individual Loligo vulgaris reynaudii off South Africa has provided insights into the daily sequence of behaviours that lead these animals to aggregate for sexual selection. Each dawn, the squid navigate for several kilometers, towards the shore, to small, well-defined zones near egg beds on the substrate. After several hours of circling above the egg beds, a pelagic, 3-D lek-like aggregation of large males forms: females are drawn in, and the aggregation condenses as the females and males pair, mate, and lay eggs. Smaller "sneaker males" remain on the periphery of the mating arena and, from this station, attempt extra-pair copulations (EPCs). The mating system of squids is thus unexpectedly complex, rivaling those of mammals and birds (2, 3). Commercial squid-jigging fishermen in South Africa have recently been attracted to the spawning grounds, and they have been successful. Moreover, their activities may be selective for large males. Thus, attention should be devoted to ensuring that such targeted fishing does not alter the characteristics of squid population genetics. Remote tracking and video observations, in combination with genetic analyses, may offer a new opportunity to monitor mating effort and reproductive success, and thus to manage the fishery.

Squid (Lolliguncula brevis) life in shallow waters: oxygen limitation of metabolism and swimming performance.

Squid (Lolliguncula brevis) were exercised in a tunnel respirometer during a stepwise increase in water velocity in order to evaluate the anaerobic threshold, i.e. the critical swimming speed above which anaerobic metabolism contributes to energy production. The average anaerobic threshold was found at speeds of 1.5-2 mantle lengths s-1. Above this velocity, alpha-glycerophosphate, succinate and octopine started to accumulate in the mantle tissue. ATP levels fell and phospho-L-arginine was progressively depleted, while the levels of glucose 6-phosphate and inorganic phosphate rose. The finding of a simultaneous onset of anaerobic metabolism in the cytosol and the mitochondria indicates that a limited oxygen supply to the mitochondria elicits anaerobic energy production. This finding is opposite to the situation found in many other vertebrate and invertebrate species, in which energy requirements in excess of aerobic energy production are covered by anaerobic metabolism, with mitochondria remaining aerobic. In L. brevis, swimming at higher speeds is associated with a small factorial increase in metabolic rate based on a high resting rate of oxygen consumption. Pressure recordings in the mantle cavity support this finding, indicating a high basal level of spontaneous activity at rest and a small rise in mean pressure at higher swimming velocity. Bursts of higher pressures from the jet support elevated swimming speeds and may explain the early transition to anaerobic energy production which occurs when pressure amplitudes exceed 1.2-1.5 kPa or when mean pressure rises above 0.22-0.25 kPa. The finding of mitochondrial hypoxia at a low critical speed in these squid is interpreted to be related to their life in shallow coastal and bay waters, which limits the necessity to maintain high swimming velocities. At increased swimming velocities, the animals oscillate between periods of high and low muscular activity. This behaviour is interpreted to reduce transport cost and to permit a longer-term net use of anaerobic resources when speed exceeds the critical value or when the squid dive into hypoxic waters. The simultaneous onset of anaerobic metabolism in the cytosol and the mitochondria emphasizes that squid generally make maximal use of available oxygen under resting conditions, when their energy requirements are the highest among marine invertebrates.

Metabolism and energetics in squid (Illex illecebrosus, Loligo pealei) during muscular fatigue and recovery.

The concentrations of intermediate and end products of anaerobic energy metabolism and of free amino acids were determined in mantle musculature and blood sampled from cannulated, unrestrained squid (Loligo pealei, Illex illecebrosus) under control conditions, after fatigue from increasing levels of exercise, and during postexercise recovery. Phosphagen depletion, accumulation of octopine (more so in Illex than in Loligo), and accumulation of succinate indicate that anaerobic metabolism contributes to energy production before fatigue. Proline was a substrate of metabolism in Loligo, as indicated by its depletion in the mantle. In both species, there was no evidence of catabolism of ATP beyond AMP. A comparison of the changes in the free and total levels of adenylates and the phosphagen indicates an earlier detrimental effect of fatigue on the energy status in Loligo. The acidosis provoked by octopine formation in Illex was demonstrated to promote the use of the phosphagen and to protect the free energy change of ATP such that the anaerobic scope of metabolism during swimming is extended and expressed more in Illex than in Loligo. In both species, there was no decrease in the sum of phospho-L-arginine, octopine, and L-arginine, and thus no release of octopine from the mantle, thereby supporting our earlier claim that octopine and associated protons are recycled in the mantle tissue. Overall, the metabolic strategy of Loligo is much less disturbing for the acid-base status. This strategy and the alternative strategy of Illex to keep acidifying protons in the tissue may be important for the protection of hemocyanin function in the two species.

Detection of canine distemper virus in bone cells in the metaphyses of distemper-infected dogs.

In the light of recent evidence implicating canine distemper virus (CDV) as a possible etiologic agent in Paget's disease of bone, we thought that it would be of interest to examine distemper-infected bone in the natural host. Samples from the long bones, spleen, and bladder of four distemper-infected and three uninfected dogs were examined for the presence of CDV nucleocapsid and phosphoprotein genes and the measles virus (MV) nucleocapsid gene using the technique of in situ hybridization with radioactively labeled riboprobes. Two of the four distemper-infected dogs showed strongly positive hybridization with both of the CDV probes. The signal was present in marrow cells, in osteoblasts, in osteocytes, and particularly in osteoclasts. No hybridization was seen over the cartilage cells of the growth plate, and there was a clear line of demarcation at the point of invasion of osteoclasts and vascularization. The spleen and bladder samples from infected dogs also showed positive hybridization. There was no hybridization with the MV probe in any of the distemper-infected tissue. Samples from the uninfected dogs showed no evidence of hybridization with either the CDV or MV probes. These results show that CDV can infect bone cells of the natural host and provide further support for the theory that CDV may play a role in human Paget's disease of bone.

Acid-base regulation in exercising squid (Illex illecebrosus, Loligo pealei).

Squid (Illex illecebrosus, Loligo pealei) were cannulated in the vena cava and swum in a Beamish-type respirometer. Gas tensions and acid-base variables as well as octopine levels were estimated in samples of the mantle and of venous blood collected from quiescent, exercised, and recovered animals. When exhausted, both species exhibited a decrease in vena cava oxygen tensions and a slight alkalosis. With high swimming speeds prior to exhaustion in Illex a slight acidosis developed in the blood, which was linked to a severe intracellular acidosis. Generally, the drop in intracellular pH was linearly correlated with octopine accumulation in this species. Metabolic proton (and end-product) release from the mantle, however, was minimal, thus protecting arterial oxygen binding. High PCO2 values in the mantle of both species lead to the conclusion that the vena cava values analyzed in this and all literature studies on unrestrained cephalopods may not reflect the scope of respiratory acid-base changes in venous blood. Although metabolic changes in blood acid-base status are negligible, the respiratory acidification of venous mantle blood may allow for a classical function of Bohr and Haldane effects in these animals.

An in vivo model system for the study of avian osteoclast recruitment and activity.

We have developed a model system for the study of osteoclast recruitment and activity using devitalized bovine cortical bone slices implanted onto the chorioallantoic membrane (CAM) of chicken embryos. Bone slices were examined after 3, 6, and 8 days of incubation on the CAM. A marked cellular reaction to the bone was observed, characterized by a prominent angiogenic response. Upon histological examination, numerous multinucleated giant cells were associated with the undersurface of the bone slice and concentrated towards its periphery. These multinucleated cells were often associated with resorption lacunae and demonstrated ruffled borders when viewed by transmission electron microscopy. Removal of the cells and examination of the bone surface by scanning electron microscopy revealed numerous resorption pits characteristic of osteoclastic activity. These pits were evident on day 3 of incubation and appeared to be more extensive by day 8. This work demonstrates that the cells recruited to such ectopically implanted devitalized bone slices are functional osteoclasts, and that this system may provide a useful model for the study of osteoclast recruitment and activity.

Location of osteoclast precursors in fetal rat calvaria cultured on collagen gels.

Although osteoclasts are derived from hematopoietic cells, the exact identity of their precursors and the mechanism for their recruitment onto bone surfaces remain unclear. We wished to study their differentiation in the fetal rat calvaria and to locate its source of osteoclast precursor cells. Osteoclasts were detected by neutral red staining or cytochemical reaction for acid phosphatase of intact bone (cell number and area measured by computerized image analysis) or in cryostat sections of bone (enzyme activity measured by quantitative cytochemistry). Histology of semithin sections of fixed bones was also examined. The 19 day calvariae contained few mature osteoclasts. After 48 h culture on gels of type 1 collagen (1.5 mg/ml) supplemented with 5 mM calcium beta-glycerophosphate, 10 mM proline, and 2 micrograms/ml ascorbic acid, numerous large osteoclasts were seen on their endocranial surfaces. In contrast, cell morphology and enzyme activity deteriorated in bones cultured in liquid medium. The cells that formed in vitro rapidly responded to calcitonin by contraction. Stripping of endocranial membranes from the calvariae prevented osteoclast formation in culture, but these cells were seen when "stripped" bones had been cocultured with their membranes for 48 h or with intact 16 day calvariae (well before the onset of osteogenesis). Few osteoclasts were found when an 0.22 micron filter was inserted between the stripped calvaria and the endocranial membranes. We conclude that the endocranial membranes, which contain the meningeal blood vessels, are a major source of osteoclast precursors and that these cells are present in calvarial tissue even before the onset of osteogenesis.

A quantitative cytochemical assay for osteoclast acid phosphatase activity in foetal rat calvaria.

Acid phosphatase activity is prominent in osteoclasts (bone resorbing cells) and has been implicated in the process of bone resorption, although its precise role is not understood. To study the distribution and activity of this enzyme, a quantitative cytochemical method has been developed using undecalcified fresh frozen sections of foetal rat calvariae. Sections were allowed to react with 3 mM naphthol ASBI phosphate at pH 5.0, and the reaction was stopped by rinsing in ice-cold tap water containing 50 mM sodium fluoride. The reaction product was post-coupled to Fast Garnet at 4 degrees C. The absorbance of areas of reaction product in the cytoplasm was measured using scanning and integrating microdensitometry. The initial velocity rate was maintained for up to 2 min at pH 5.0 with a substrate concentration of 3 mM and a section thickness of 5 micron. Under these conditions reaction product was localized to osteoclasts and the surface of bone matrix beneath these cells. Activities in osteoblasts and chondrocytes were negligible. Osteoclastic acid phosphatase was almost totally inhibited by 10 mM fluoride and reduced by 70% by 100 mM tartrate.