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The availability of animal models with disrupted genes has increased the need for small-scale measurement devices. Recently, we developed an experimental device to assess in situ mechanical properties of isometric contractions of intact muscle complexes of the mouse. Although this apparatus provides valuable information on muscle mechanical performance, it is not appropriate for determining contractile properties during shortening and lengthening contractions. In the present study we therefore developed and evaluated an experimental apparatus for assessment of shortening and lengthening contractile properties of intact plantar and dorsal flexors of the mouse. The current through a custom-built, low-inertia servomotor was measured to assess contractile muscular torque ranging from -50 to mN.m. Evaluation of the fixation procedure of the animal to the apparatus via 3-D monitoring of the muscle-tendon complex length showed that the additional shortening in length due to a contraction with maximal torque output has only minor effects on the measured torque. Furthermore, misalignment of the axis of rotation of the apparatus relative to the axis of rotation in the ankle joint, i.e. eccentricity, during a routine experiment was estimated to be less than 1.0 mm and hence did not influence the measured torque output under our experimental conditions. Peak power per unit muscle mass (mean +/- SD) of intact dorsal and plantar flexors was 0.27 +/- 0.02 and 0.19 +/- 0.03 W.g-1, respectively. The angular velocity at maximal peak power generated by the dorsal flexor complex and the plantar flexor complex was 1100 +/- 190 and 700 +/- 90 degrees.s-1, respectively.

A mixing pump that creates an accurate mixture of three gases at predetermined fractional ratios that can be set in steps of 10 ppm is described. A nearly continuous flow of each of the three component gases is produced by pistons driven by stepping motors; the gas mixture is forwarded by a fourth piston. The flow of each component gas is adjusted by the stepping frequency of the motor and a microcomputer system is used to adjust the three frequencies according to the desired fractional concentrations. The total flow of the gas mixture is adjustable between 0.1-500 ml/min and is nearly independent of the after-load. The accuracy of the pump was tested by mixing the respiratory gases, O2 and CO2, with various carrier gases (N, He or Ar) at various fractional ratios and total flow rates. The fractions of O2 and CO2 in the mixture were analysed with the Scholander technique. In the physiological range, the mixing error in the gas fractions was less than 4%. The pump is, thus, suited for producing calibration mixtures.

A method of calibrating photodiode measurements of cell motion is described. A transparent glass plane is inserted perpendicular to the light path, in front of the photodiode. When the glass plane is rotated, changing the angle between the plane and the light path, the image is displaced. The rotatable glass plane acts as an optical lever to simulate cell motion. The optical lever is easy to construct and offers excellent linearity and accuracy.

Short duration washin and washout experiments were carried out with the gases ether (diethyl ether), ethyl acetate and acetone in order to study the excretion behaviour in the lung of gases highly soluble in blood and tissues. Carbon dioxide (CO2) was analyzed as a reference gas during the washout. The excretion values for acetone were generally the lowest, those for ether the highest, and intermediate ones were obtained for ethyl acetate. These results are in accord with the experimental data of others. Analysis of the washout curves shows that for acetone the volume of gas expired before the beginning of phase II is considerably smaller than found for CO2 and ether. During washout, the slope of the alveolar plateau for acetone is negative for the first few breaths and becomes positive thereafter; however, it remains lower than the slopes for CO2 and ether, which are always positive. These two phenomena occurring during washout clearly demonstrate that the acetone in the expired air must originate from the epithelial tissue lining the conducting airways. We conclude, therefore, that, in terms of gas transport, the conducting airways behave differently for poorly and highly soluble gases and this provides a physiological basis for the deviating excretion behaviour of highly soluble gases.

The transients of the tension and the angle of a first order diffraction line of isolated single intact muscle fibres of the M. lumbricalis IV of the foot resulting from rapid length changes have been measured. Furthermore simulations of the tension transients, taking the resonance frequency of the force transducer, the inertia of the fibre and the influence of the surrounding fluid into account, are presented. The tension transients could be simulated by a system of continuous elements with undamped elasticity - Young's modulus E1 = 1.8 X 10(8) N/m2 - in series with elasticity E2 = 5.4 X 10(7) N/m2 parallel to a damping element - coefficient of damping mu = 2,300 Ns/m2; yielding a relaxation time of 10 mus. The Young's modulus of the undamped series elasticity E1 implies that a sudden shortening of 2 nm per half sarcomere should reduce the active tension to zero. The results obtained from the diffraction pattern recording show that the displacement in the fibre is in accordance with the relative length change caused by the displacement generator. It is therefore believed that the tendon and tendon-muscle junction are sufficiently stiff to allow ascribing the above-mentioned elasticity and damping to the fibre itself.

Pressure-diameter curves were determined on mesometrial (uterine radial) arteries obtained from guinea pigs during different stages of ovarian cycle and pregnancy. In order to investigate changes in arterial diameter that are not caused by the vascular musculature, we studied excised arteries that were relaxed by papaverine (40 mg/l). The pressure-diameter curves were found to be shifted toward wider diameters when the arteries studied were obtained from pregnant, oestrous or oestrogen treated animals (oestradiol benzoate, 10 micrograms s.c.): the external diameter at 60 mm Hg transmural pressure on the 14th day of pregnancy exceeded the one during dioestrus by 50%. At term, there was a 7-fold increase. During oestrus and 24 h after oestrogen treatment the diameter was 30-40% wider than during dioestrus. Moreover, we found that the diameter measured on excised vessels in the presence of papaverine is the same as the one observed in situ. We conclude that the mesometrial arteries of guinea pigs possess a muscle-independent mechanism of lumen regulation. The mechanism operates in non-pregnant and pregnant animals. It may be induced by oestrogen and seems to be qualified for long-term lumen regulation during pregnancy. In contrast, dilation by smooth muscle relaxation appears to be of minor importance in mesometrial arteries.

A method permitting rapid and frequent determination of the total ionic concentration in renal tubular fluid samples, by measuring sample electrical impedance, is described. No special electrodes or sophisticated equipment are required. Since picoliter volumes are sufficient for measurement, tubular fluid dynamics are not significantly disturbed by the sampling procedure. The sample is aspirated into the tip of the standard glass micropipette and its ionic concentration, in the range of 100-1000 mmol/l, can be determined by any sensitive impedance-measuring device. In the present studies the Wheatstone bridge of the Wiederhielm apparatus for pressure measurement in micropuncture studies was used for the purpose. The obvious limitation of the method is that the measurements do not include non-electrolytes and cannot differentiate between individual ion species.

A micro cuvette is described, which allows to measure mechanical, optical and energetic parameters of single skinned muscle fibres or small fibre bundles. Two examples for possible applications are given: The measurement of ATPase activity of single muscle fibres. The detection of fluorescence changes due to Ca binding to fluorescence labeled TnC, which was incorporated into small bundles of rabbit psoas fibres before the experiment.

In ten normal subjects, series dead space was determined for six intravenously infused inert gases (SF6, ethane, cyclopropane, fluothane, ether, acetone) from their expired and alveolar concentrations. The method for sampling alveolar gas was based on the criterion of identity of mean alveolar and expired gas exchange ratios. Inert gases were analysed chromatographically. Acetone, the most soluble gas, yielded the lowest dead space, the difference to the other gases being about 4.5%. This is probably due to the non-infinite value of the series dead space ventilation-perfusion ratio (VA/Q) which was estimated at about 2,000. The diffusivity, inversely related to the molecular weight, also played a role, the heaviest gas (fluothane) having a greater dead space than the lightest (ethane). The underestimation of the dead space from acetone is expected to be greater in subjects with low tidal volume and high bronchial blood flow, i. e. in some patients with respiratory disease.

An optical system for measuring changes in cell length during unloaded contractions of cardiac myocytes is described. A one-dimensional video "image" of a cell is obtained every 4 ms with a linear photodiode array, which is aligned with the longitudinal axis of the cell. The circuit used to process the image from the photodiode array has a variety of features to aid in the accurate determination of the distance between the ends of the cell, i.e. the cell length. First, the video image of the cell is divided into two "windows", one encompassing the "front" edge of the cell, the other encompassing the "rear" edge. Other cells or debris beyond the cell edges are excluded. Changes in the general light level, for example as a result of debris floating above the cell, have little effect because within the windows the "background light level" is subtracted from the signals before they are processed further. To detect the cell edges, the system determines when the signals within the windows exceed (front edge) or drop below (rear edge) chosen thresholds, which are different for the front and rear edges. The system has "memory" and it identifies the rear edge of the cell as the last time the signal falls below the threshold; because of this "bright spots" within the cell are not mistaken for the end of the cell. The system has "hysteresis", which enables it to ignore small fluctuations in brightness around the threshold. The system is easy to use, accurate, readily calibrated, and it has good spatial and time resolution (about 0.25 micron and 4 ms respectively).

The current contributions of individual ionic channels can be measured by electrically isolating a small patch of membrane. To do this, the tip of a small pipette is brought into close contact with an enzymatically cleaned membrane of a hypersensitive amphibian or mammalian muscle fiber. Current flowing through the pipette is measured. If the pipette contains cholinergic agonist at mu-molar concentrations, square pulse current waveforms can be observed which represent the activation of individual acetylcholine-receptor channels. The square pulses have amplitudes of 1 to 3 pA and durations of 10--100ms. In order to obtain the necessary resolution, a delicate compromise had to be found between different experimental parameters. Pipettes with 1--3 micrometer internal diameter and a steep final taper had to be used, extensive enzyme treatment was necessary, and conditions had be to found in which channels open at a relatively low frequency.

A simple and cheap method is devised for controlling the fluid level in an open chamber for physiological or other experiments. Fluid is pumped into the chamber at variable flow rate using a conventional pump. Autoregulated drainage is achieved through a capillary tube whose bevelled opening is covered with a net with fine meshes. Part of the opening dips into the fluid surface. The correct amount of fluid is sucked out of the chamber by a constant negative pressure at the back of the tube. The 1 mm depth of a 2 microliters volume stays within +/- 2% when the normal flow of 1 microliters/s is doubled and halved, respectively. A 100-fold increase of flow, as needed for flushing the chamber, causes a rise of the surface level by 0.5 mm only.

A method is described here for making multiple fast external solution changes at the tip of a patch pipette. The time for the change, 0.2 ms, has been established by measuring changes in liquid junction potential at the tip of an open patch pipette. This technique of producing an abrupt change in solution allows agonist/receptor reactions to be studied under non-equilibrium conditions. We have applied this technique to the nicotinic receptors in outside-out patches from skeletal muscle cell line C2 (Jaffe and Saxel 1979) and from bovine adrenal chromaffin cells. The application of step changes in acetylcholine concentration produces current traces with a characteristic shape, which may be compared with the predictions of established models for the activation and desensitisation of the nicotinic receptor. The results of making single steps and also short pulses in acetylcholine concentration are demonstrated. The direct comparison of two different cholinergic agonists is demonstrated.

The construction of a new type of vascular cuff occluder for use in conscious animals is described. This occluder has the following features: 1. Safe complete vessel occlusion. 2. Occlusion at a definite cuff pressure with extrathoracally pressure control behind the cuff. 3. Long term implantability. 4. Possibility of multiple usage and sterilization.

In order to determine the tension response of a stimulated intact muscle fibre to a very fast length change a servo system has been developed capable of imposing a displacement of 50 micrometers i.e. about 1% of the fibre length, within 40 microseconds. The system is designed so as to limit the displacement overshoot to less then 1%. This paper presents the performance of the system: both unloaded and loaded with a muscle fibre.

An instrument and technique have been devised for the production of complete atrioventricular heart block in anesthetized, closed-chest animals. The instrument is inserted into the right atrium via the right external jugular vein. Localization of the atrioventricular node region by the visualization of intracardiac landmarks with fluoroscopy makes electrical recording unnecessary. Discrete injections of a radiopaque necrosing solution produces the heart block. Procedure time is under 30 min a success rate of 90%.

Tympanic membrane temperature (Ttymp) and deep esophageal temperature (Tes) were measured in 8 subjects during normal outdoor bicycling. Metabolic rate (VO2) was determined by the Douglas bag method. Heart rate was sampled continuously. Skin surface temperatures were measured at the forehead, chest and shoulder, and core temperatures in the deep esophagus and at the tympanic membrane using a radio telemetry system. For each outdoor experiment an indoor experiment in a climatic chamber, adjusted to the same air temperature but in still air, was performed. The subjects exercised at the same VO2 as in the outdoor trial on a stationary bicycle ergometer. Measurements were taken with the same equipment as in the outdoor experiments. O2-consumption (l.min-1) and heart rates (beats.min-1) were similar during outdoor and indoor bicycling, averaging 2.38 +/- 0.018 (SE) and 2.26 +/- 0.07, 141 +/- 7 and 147 +/- 8, respectively. During steady state Tes was the same during outdoor and indoor bicycling (37.95 degrees C), while Ttymp was significantly lower during outdoor bicycling. delta (Tes-Ttymp) was 1.25 degrees C during outdoor and 0.5 degrees C during indoor exercise. It is concluded that, if tympanic temperature is lowered by counter-current cooling of its arterial supply, then cooling of the brain may also take place in humans during physical activity under normal outdoor conditions with convective air movements. But the magnitude of a possible brain cooling cannot be deduced from the fall in tympanic temperature.

The construction of a microscope stage temperature controller is described that works equally well with upright or inverted microscopes. The control circuit directly regulates the bath temperature near the physiological preparation from 0.0 degree C to 40.0 degrees C with a stability of +/- 0.1 degrees C; it allows biophysical studies of whole-cell or single channel currents to be carried out at different temperatures without introducing additional electrical noise into the measurements. The device may also be of interest to neurobiologists who work with tissue-culture preparations.