Fluorescence anisotropy from diphenylhexatriene in rat liver plasma membranes

Rat livers were fractionated to obtain intracellular membrane preparations and a highly purified preparation of bile canaliculi. The fraction containing bile canaliculi was homogenized and subfractionated to give fractions representing fragments of contiguous membrane and of canalicular microvilli. The relative purity and extent of contamination of each preparation was determined. When the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene was incorporated into aliquots of each fraction at the same probe: lipid ratio and the steady-state anisotropy of its fluorescence measured, it was found that the plasma membrane preparations were much more ordered than the intracellular membrane preparations. Of the plasma membrane preparations, that containing the canalicular microvilli was the most ordered, even allowing for any contribution of contaminants. Thus the microvillus membrane of the bile canaliculus appears to be the most ordered domain of the plasma membrane of the hepatocyte. The high order in this domain may be a factor in reducing the susceptibility to bile salt damage during bile secretion, since it is this region which is exposed to high concentrations of bile salts in vivo.     

Burning the engine: a time-marching computation of fat and protein consumption in a 5420-km non-stop flight by great knots, Calidris tenuirostris

Samples of great knots (Calidris tenuirostris) were collected in an earlier project, before and after a 5420‐km migration stage from Australia to China (believed to be flown non‐stop) to determine the mass of fat consumed, and also the mass of protein withdrawn from the flight muscles and other organs. The flight was simulated by a “time‐marching” computation, which calculated the fuel energy required, and allowed different hypotheses to be tried for the consumption of protein. The simulation predicted that the great knots would take about 4 days to cover the distance, in agreement with field estimates. Realistic predictions of the consumption of fat and protein were obtained by setting the conversion efficiency to 0.23 and the body drag coefficient to 0.10, withdrawing sufficient protein from the flight muscles to keep the specific work in the myofibrils constant throughout the flight, and taking enough additional protein from other tissues to bring the energy derived from oxidising protein to 5% of the total energy consumed. The same computation was applied to published data on the pre‐migration body composition of bar‐tailed godwits (Limosa lapponica), which are said to migrate over 10 000 km from Alaska to New Zealand. The computed range for a sample killed by collision with an obstruction, while actually departing from Alaska, was sufficient to reach the South Pole. A second sample, shot before departure from New Zealand, would have run out of fat before reaching Alaska, but could easily have reached northern Australia, where these godwits stage on their northbound migration. The higher range estimate for the Alaskan birds was not due to higher fat mass (only 5% difference) but to a higher fat fraction, which they had achieved by reducing the mass of other organs before departure. Some recent observations of high chemical power, observed in wind tunnel experiments, have been interpreted as being due to much lower conversion efficiency than the value of 0.23 assumed here, but this interpretation is flawed. Measurements of mechanical power from another wind tunnel project were also unexpectedly high, suggesting that unsteady flight by wind tunnel birds increases their power requirements, both mechanical and chemical, with no implications for efficiency. The calculated power is for “steady horizontal flight”, meaning that a valid test of predicted power requires birds to be trained to hold a constant position in the test section, while maintaining a steady wingbeat frequency and amplitude. This has not been achieved in recent experiments, and is hard to achieve when using physiological methods, because of the long periods of continuous flight needed. Measurements of mechanical rather than chemical power require shorter flight times, and offer better prospects for reliable power measurements.     

Interspecific dominance and the foraging behaviour of juncos

We observed mixed groups of dark-eyed juncos (Junco hyemalis) and grey-headed juncos (Junco caniceps) at baited sites in northern Arizona during the non-breeding season. In interspecific and inter-racial conflicts, J. caniceps dorsalis was dominant to J. caniceps caniceps and to two races of dark-eyed juncos. Junco caniceps dorsalis also fed significantly faster than any of the other juncos. For both species, feeding rates were approximately the same in large and small mixed-species groups, though in larger groups, individual grey-headed juncos won conflicts at a higher rate and individual dark-eyed juncos lost conflicts at a higher rate. Also, dark-eyed juncos fed at a significantly lower rate in groups comprised mostly of grey-headed juncos than in groups of similar size but composed mostly of conspecifics. Residency times and recapture probabilities were similar for the two species, suggesting little difference in over-winter survival.     

Maternal or fetal origin of rhesus monkey (Macaca mulatta) amniotic fluid leukocytes can be identified by polymerase chain reaction using the zinc finger Y gene.

Leukocytes can be found in substantial numbers within the intrauterine tissues and amniotic fluid of women, and play a central role in the pathophysiology of infection-related preterm labor by their production of proinflammatory mediators. It remains unclear whether these leukocytes represent a fetal immune response, a maternal response, or a combination of the two. The objective of this study was to develop a test in the rhesus monkey (Macaca mulatta) suitable for determining the percentage of male fetal cells present in a population of leukocytes recovered from blood or amniotic fluid. We found inadequate specificity for rhesus monkey cells using commercial human Y-chromosome paint kits (fluorescence in situ hybridization (FISH)). Human-specific primers for the repetitive Y chromosome DYZ-1 locus employed in the polymerase chain reaction (PCR) produced an unacceptable percentage of false positives. However, we successfully developed a PCR-based test using rhesus-specific primers for the zinc finger Y (ZFY) locus. Densitometry of PCR products from known ratios of male and female adult peripheral leukocytes generated a linear standard curve which provided quantitative results and required only 400 cells per sample. The rhesus beta globin (RBG) gene served as an internal control. The PCR test correctly discriminated the sex of peripheral leukocytes in 20 adult males, 20 adult females, two male fetuses, and one female fetus. Serial samples of amniotic fluid from four chronically catheterized rhesus monkeys bearing male fetuses were used to confirm the utility of this assay for quantifying fetal cells in amniotic fluid. In conclusion, we have developed a PCR test which is suitable for distinguishing male from female cells in adult and fetal blood and in amniotic fluid, which lends itself to a variety of diagnostic and biologic applications in the rhesus monkey and potentially in other nonhuman primates.     

Parasympathetic innervation of canine tracheal smooth muscle

To investigate whether efferent parasympathetic fibers to the trachealsmooth muscle course through the pararecurrent nerve rather than therecurrent or the superior laryngeal nerve, we stimulated all threenerves in anesthetized dogs. We also recorded the pararecurrentnerve activity response to bronchoconstrictor stimuli and compared itwith pressure changes inside a saline-filled cuff of an endotrachealtube. Electrical stimulation (30 s, 100 Hz, 0.1 ms, 10 mA) increasedtracheal cuff pressure by 21.0 ± 3.2 and 1.3 ± 0.7 cmH₂O for the pararecurrent and the recurrent laryngealnerve, respectively. Stimulation of the superior laryngeal nerveincreased tracheal cuff pressure before, but not after, sectioning ofthe ramus anastomoticus, which connects it to the pararecurrent nerve.Intravenous administration of sodium cyanide increased pararecurrentnerve activity by 208 ± 51% and tracheal cuff pressure by14.4 ± 3.5 cmH₂O. Elevation of end-tidalPCO₂ to 50 Torr increased pararecurrent nerveactivity by 49 ± 19% and tracheal cuff pressure by 8.4 ± 3.6 cmH₂O. Further elevation to 60 Torr increasedpararecurrent nerve activity by 101 ± 33% and tracheal cuffpressure by 11.3 ± 2.9 cmH₂O. These results lead usto the conclusion that parasympathetic efferent fibers reach the smoothmuscle of the canine trachea via the pararecurrent nerve.