Myocardial twitch duration and the dependence of oxygen consumption on pressure-volume area: experiments and modelling.

We tested the proposition that linear length dependence of twitch duration underlies the well-characterised linear dependence of oxygen consumption (V(O(2)) ) on pressure­volume area (PVA) in the heart. By way of experimental simplification, we reduced the problem from three dimensions to one by substituting cardiac trabeculae for the classically investigated whole-heart. This allowed adoption of stress­length area (SLA) as a surrogate for PVA, and heat as a proxy for V(O(2)) . Heat and stress (force per cross-sectional area), at a range of muscle lengths and at both 1 mM and 2 mM [Ca(2+)](o), were recorded from continuously superfused rat right-ventricular trabeculae undergoing fixed-end contractions. The heat­SLA relations of trabeculae (reported here, for the first time) are linear. Twitch duration increases monotonically (but not strictly linearly) with muscle length. We probed the cellular mechanisms of this phenomenon by determining: (i) the length dependence of the duration of the Ca(2+) transient, (ii) the length dependence of the rate of force redevelopment following a length impulse (an index of Ca(2+) binding to troponin-C), (iii) the effect on the simulated time course of the twitch of progressive deletion of length and Ca(2+)-dependent mechanisms of crossbridge cooperativity, using a detailed mathematical model of the crossbridge cycle, and (iv) the conditions required to achieve these multiple length dependencies, using a greatly simplified model of twitch mechano-energetics. From the results of these four independent investigations, we infer that the linearity of the heat­SLA relation (and, by analogy, the V(O(2))­PVA relation) is remarkably robust in the face of departures from linearity of length-dependent twitch duration.

Assessment of dispersion of airborne particles of oral/nasal fluid by high flow nasal cannula therapy.

BACKGROUND: Nasal High Flow (NHF) therapy delivers flows of heated humidified gases up to 60 LPM (litres per minute) via a nasal cannula. Particles of oral/nasal fluid released by patients undergoing NHF therapy may pose a cross-infection risk, which is a potential concern for treating COVID-19 patients. METHODS: Liquid particles within the exhaled breath of healthy participants were measured with two protocols: (1) high speed camera imaging and counting exhaled particles under high magnification (6 participants) and (2) measuring the deposition of a chemical marker (riboflavin-5-monophosphate) at a distance of 100 and 500 mm on filter papers through which air was drawn (10 participants). The filter papers were assayed with HPLC. Breathing conditions tested included quiet (resting) breathing and vigorous breathing (which here means nasal snorting, voluntary coughing and voluntary sneezing). Unsupported (natural) breathing and NHF at 30 and 60 LPM were compared. RESULTS: Imaging: During quiet breathing, no particles were recorded with unsupported breathing or 30 LPM NHF (detection limit for single particles 33 µm). Particles were detected from 2 of 6 participants at 60 LPM quiet breathing at approximately 10% of the rate caused by unsupported vigorous breathing. Unsupported vigorous breathing released the greatest numbers of particles. Vigorous breathing with NHF at 60 LPM, released half the number of particles compared to vigorous breathing without NHF.Chemical marker tests: No oral/nasal fluid was detected in quiet breathing without NHF (detection limit 0.28 µL/m3). In quiet breathing with NHF at 60 LPM, small quantities were detected in 4 out of 29 quiet breathing tests, not exceeding 17 µL/m3. Vigorous breathing released 200-1000 times more fluid than the quiet breathing with NHF. The quantities detected in vigorous breathing were similar whether using NHF or not. CONCLUSION: During quiet breathing, 60 LPM NHF therapy may cause oral/nasal fluid to be released as particles, at levels of tens of µL per cubic metre of air. Vigorous breathing (snort, cough or sneeze) releases 200 to 1000 times more oral/nasal fluid than quiet breathing (p < 0.001 with both imaging and chemical marker methods). During vigorous breathing, 60 LPM NHF therapy caused no statistically significant difference in the quantity of oral/nasal fluid released compared to unsupported breathing. NHF use does not increase the risk of dispersing infectious aerosols above the risk of unsupported vigorous breathing. Standard infection prevention and control measures should apply when dealing with a patient who has an acute respiratory infection, independent of which, if any, respiratory support is being used. CLINICAL TRIAL REGISTRATION: ACTRN12614000924651.

Ultrasonographic assessment of the rate of solid-phase gastric emptying in dogs.

Twelve healthy dogs were used in an ultrasonographic assessment of the effect of the composition of a solid meal on the rate of gastric emptying. The dogs were fasted for nine hours before they were fed either a standard or a high energy density test meal, in a cross-over study design. The gastric antrum was visualised with a 6.5 MHz microconvex transducer, and the area inside the elliptical shape defined by the craniocaudal and ventrodorsal diameters of the stomach was measured. Antral images were acquired at regular intervals for six hours after the ingestion of the test meal. Three indices to describe the rate of gastric emptying were computed: the gastric half-emptying time (t1/2), the time to 50 per cent maximal antral area (t50%), and the time of maximal antral area (tmax). The values of t50% and t1/2 calculated for the high energy density meals were significantly longer than for the standard meals, but there was no significant difference between the tmax values.

Effects of BDM, [Ca2+]o, and temperature on the dynamic stiffness of quiescent cardiac trabeculae from rat.

Studies of the passive mechanical properties of cardiac tissue have traditionally been conducted at subphysiological temperatures and various concentrations of extracellular Ca(2+) ([Ca(2+)](o)). More recently, the negative inotropic agent 2,3-butanedione monoxime (BDM) has been used. However, there remains a lack of data regarding the influence of temperature, Ca(2+), and BDM on the passive mechanical properties of cardiac tissue. We have used the dynamic stiffness technique, a sensitive measurement of cross-bridge activity, in which minute (approximately 0.2% of muscle length) sinusoidal perturbations are applied at various frequencies (0.2-100 Hz) to quiescent, viable right ventricular rat trabeculae at two temperatures (20 degrees C and 26 degrees C) and at two [Ca(2+)](o) (0.5 and 1.25 mM) in the presence and absence of BDM (20 mM). The stiffness spectra (amplitude and phase) were sensitive to temperature and [Ca(2+)](o) in the absence of BDM but insensitive in the presence of BDM. From the index of cross-bridge cycling (the ratio of high- to low-frequency stiffness amplitude), we infer that BDM inhibits a small degree of spontaneous sarcomere activity, thereby allowing the true passive properties of trabeculae to be determined. In the absence of BDM, the extent of spontaneous sarcomere activity decreases with increasing temperature. We caution that the measured mechanical properties of passive cardiac tissue are critically dependent on the experimental conditions under which they are measured. Experiments must be performed at sufficiently high temperatures (>25 degrees C) to ensure a low resting concentration of intracellular Ca(2+) or in the presence of an inhibitor of cross-bridge cycling.

Sensitivity of mitomycin-C treated McCoy cells for isolation of Chlamydia trachomatis from genital specimens.

A new cell culture method was developed for the diagnostic isolation of Chlamydia trachomatis using McCoy cells pretreated with mitomycin-C. This drug acts on the cell by producing cross links between strands of DNA and results in the production of large, flat cells, which are similar to irradiated cells, and large chlamydial inclusions. In tests using a laboratory isolate of Chlamydia trachomatis, there were significantly more inclusions produced in mitomycin-treated monolayers than in cycloheximide-treated monolayers. Using clinical specimens, significantly more isolates were obtained in mitomycin-treated cells than in cycloheximide-treated cells. The new cell culture technique therefore offers a cell culture method which has the advantages of producing large inclusions similar to irradiated cells but with the convenience of drug treatment.

A sensitive flow-through microcalorimeter for measuring the heat production of cardiac trabeculae.

Measurement of the energy consumption of isolated cardiac muscle requires a flow-through microcalorimeter with sensitivity in the microW range. In this paper we describe and characterize a sensitive flow-through microcalorimeter, designed and constructed for measuring the heat output of cardiac trabeculae. The device exploits a non-contact, temperature-sensing technique utilizing infra-red-sensitive, thin-film thermopile sensors. The microcalorimeter achieves a sensitivity of 1.8-1.9 V/W at a flow rate of 1 microl/s, with a time constant of approximately 3.5 s. The typical power signal-to-noise ratio is better than 200. Predictions of a finite element model of the calorimeter's characteristics compare favourably with measured data.

Strain softening is not present during axial extensions of rat intact right ventricular trabeculae in the presence or absence of 2,3-butanedione monoxime.

Recent studies of passive myocardial mechanics have shown that strain softening behavior is present during both inflation of isolated whole rat hearts and shearing of tissue blocks taken from the left ventricular free wall in pigs. Strain softening is typically manifested by a stiffer force-extension relation in the first deformation cycle relative to subsequent cycles and is distinguished from viscoelasticity by a lack of recovery of stiffness, even after several hours of rest. The causes of this behaviour are unknown. We investigated whether strain softening is observed in uniaxial extensions of intact, viable, rat right ventricular (RV) cardiac trabeculae. Stretch and release cycles of 5%, 10%, and 15% muscle length were applied at a constant velocity at 26 degrees C. Muscles were tested in random order in the presence and absence of 50 mM 2,3-butanedione monoxime (BDM). Whereas strain softening was displayed by nonviable trabeculae, it was not observed in viable preparations undergoing physiologically relevant extensions whether in the presence or absence of BDM. BDM also had no effect on passive compliance. There was a reversible increase of muscle compliance between the first and subsequent cycles, with recovery after 30 s of rest, independent of the presence of BDM. We conclude that strain softening is neither intrinsic to viable rat RV trabeculae nor influenced by BDM and that passive trabeculae compliance is not altered by the addition of BDM.

Strain softening behaviour in nonviable rat right-ventricular trabeculae, in the presence and the absence of butanedione monoxime.

Strain softening is commonly reported during mechanical testing of passive whole hearts. It is typically manifested as a stiffer force-extension relationship in the first deformation cycle relative to subsequent cycles and is distinguished from viscoelasticity by a lack of recovery of stiffness, even after several hours of rest. The cause of this behaviour is presently unknown. In order to investigate its origins, we have subjected trabeculae to physiologically realistic extensions (5-15% of muscle length at 26 degrees C and 0.5 mm Ca(2+)), while measuring passive force and dynamic stiffness. While we did not observe strain softening in viable trabeculae, we found that it was readily apparent in nonviable (electrically inexcitable) trabeculae undergoing the same extensions. This result was obtained in both the presence and absence of 2,3-butanedione monoxime (BDM). Furthermore, BDM had no effect on the passive compliance of viable specimens, while its presence partly inhibited, but could not prevent, stiffening of nonviable specimens. Loss of viability was accompanied by a uniform increase of dynamic stiffness over all frequencies examined (0.2-100 Hz). The presence of strain softening during length extensions of nonviable tissue resulted in a comparable uniform decrease of dynamic stiffness. It is therefore concluded that strain softening is neither intrinsic to viable rat right ventricular trabeculae nor influenced by BDM but, rather, reflects irreversible damage of tissue in partial, or full, rigor.