The interaction of the upper airway and thermo-metabolism on respiratory rhythm during non-REM sleep in the developing lamb.

This study investigates the role of metabolic rate and of vagal airway mechanisms in sustaining rhythmic breathing in the developing lamb. Fifteen lambs were prepared, at 2 days of age under fluothane anaesthesia, for sequential studies at 4, 14, 30, 45, and 55 days of age. At each age they were maintained at an ambient temperature of 5, 10, 15, 20, 25 and 30 degrees C for at least one hour before measurements were made during N-REM sleep. In 6 lambs at 4 days and in all lambs at older ages the upper airway was by-passed (by opening a tracheal window) for 10-15 minutes at each ambient temperature. Oxygen consumption was unaffected by upper airway by-pass and there were no consistent changes in mean breathing frequency or amplitude, with the exception of shifts to panting at warm ambient temperatures. Breathing pattern was unaffected by upper airway by-pass in lambs at 4 days of age, but at older ages loss of regularity of breathing frequently occurred (up to 47% of 30 days-old lambs at 25 degrees C). This was related to the fall in oxygen consumption with age and to basal values at thermoneutrality, and coincided with lower respiratory rates and increased use of expiratory laryngeal braking. Periodic breathing (and apnea) of a fixed cycle length (9.3 +/- 0.36 s) was a common feature (62%) of the observed breathing dysrhythmia. In young lambs high metabolic rate sustained high frequency rhythmic breathing which was unaffected by upper airway by-pass.(ABSTRACT TRUNCATED AT 250 WORDS)     

Reconstructing muscle activation during normal walking: a comparison of symbolic and connectionist machine learning techniques

One symbolic (rule-based inductive learning) and one connectionist (neural network) machine learning technique were used to reconstruct muscle activation patterns from kinematic data measured during normal human walking at several speeds. The activation patterns (or desired outputs) consisted of surface electromyographic (EMG) signals from the semitendinosus and vastus medialis muscles. The inputs consisted of flexion and extension angles measured at the hip and knee of the ipsilateral leg, their first and second derivatives, and bilateral foot contact information. The training set consisted of data from six trials, at two different speeds. The testing set consisted of data from two additional trials (one at each speed), which were not in the training set. It was possible to reconstruct the muscular activation at both speeds using both techniques. Timing of the reconstructed signals was accurate. The integrated value of the activation bursts was less accurate. The neural network gave a continuous output, whereas the rule-based inductive learning rule tree gave a quantised activation level. The advantage of rule-based inductive learning was that the rules used were both explicit and comprehensible, whilst the rules used by the neural network were implicit within its structure and not easily comprehended. The neural network was able to reconstruct the activation patterns of both muscles from one network, whereas two separate rule sets were needed for the rule-based technique. It is concluded that machine learning techniques, in comparison to explicit inverse muscular skeletal models, show good promise in modelling nearly cyclic movements such as locomotion at varying walking speeds. However, they do not provide insight into the biomechanics of the system, because they are not based on the biomechanical structure of the system.     

Growth of separated and recombined neonatal rat uterine luminal epithelium and stroma on extracellular matrix: Effects of in vivo tamoxifen exposure

Summary We have developed a system for serum-free culture of separated uterine epithelium and stroma from 11-day-old rats recombined on extracellular matrix extracted from Englebreth-Holm-Swarm tumors. Epithelium grew and, after 2 days in culture, developed into luminal epithelial spheres (LES) surrounding a fluid-filled lumen. Individual LES cells maintained epithelial cell characteristics such as basally located nuclei, apical microvilli (oriented toward the lumen), lateral membranes with interdigitations and desmosomes, secretory Golgi complexes, and abundant mitochondria and rough endoplasmic reticulum. Secretory vesicles were ubiquitous throughout the luminal fluid. Addition of 17β-estradiol to the growth medium increased the number and longevity of the LES. Prior exposure of uteri to tamoxifen via s.c. injection in vivo on postnatal Days 1 to 5 reduced or completely inhibited formation of LES in vitro. These effects occurred regardless of whether the stromal or epithelial component of the recombinant tissue was exposed to tamoxifen. These data suggest a directive property of neonatal stroma in culture resulting in the formation of highly secretory spherical epithelial structures completely enclosing a lumen. LES formation is responsive to both estrogen (positive response) and antiestrogen (negative response).     

Reduction of ribulose biphosphate carboxylase activase levels in tobacco (Nicotiana tabacum) by antisense RNA reduces ribulose biphosphate carboxylase carbamylation and impairs photosynthesis.

The in vivo activity of ribulose-1,5-biphosphate carboxylase/oxygenase (Rubisco) is modulated in response to light intensity by carbamylation of the active site and by the binding of sugar phosphate inhibitors such as 2'-carboxyarabinitol-1-phosphate (CA 1P). These changes are influenced by the regulatory protein Rubisco activase, which facilitates the release of sugar phosphates from Rubisco's catalytic site. Activase levels in Nicotiana tabacum were reduced by transformation with an antisense gene directed against the mRNA for Rubisco activase. Activase-deficient plants were photosynthetically impaired, and their Rubisco carbamylation levels declined upon illumination. Such plants needed high CO2 concentrations to sustain reasonable growth rates, but the level of carbamylation was not increased by high CO2. The antisense plants had, on average, approximately twice as much Rubisco as the control plants. The maximum catalytic turnover rate (k cat) of Rubisco decreases in darkened tobacco leaves because of the binding of CA 1P. The dark-to-light increase in k cat that accompanies CA 1P release occurred to similar extents in antisense and control plants, indicating that normal levels of activase were not essential for CA 1P release from Rubisco in the antisense plants. However, CA 1P was released in the antisense plants at less than one-quarter of the rate that it was released in the control plants, indicating a role for activase in accelerating the release of CA 1P.     

Amino-terminal truncations of the ribulose-bisphosphate carboxylase small subunit influence catalysis and subunit interactions.

The first 20 residues at the amino terminus of the small subunit of spinach ribulose-1,5-bisphosphate carboxylase form an irregular arm that makes extensive contacts with the large subunit and also with another small subunit (S. Knight, I. Andersson, and C.-I. Brändén [1990] J Mol Biol 215: 113-160). The influence of these contacts on subunit binding and, indirectly, on catalysis was investigated by constructing truncations from the amino terminus of the small subunit of the highly homologous enzyme from Synechococcus PCC 6301 expressed in Escherichia coli. Removal of the first six residues (and thus the region of contact with a neighboring small subunit) affected neither the affinity with which the small subunits bound to the large subunits nor the catalytic properties of the assembled holoenzyme. Extending the truncation to include the first 12 residues (which encroaches into a highly conserved region that interacts with the large subunit) also did not weaken intersubunit binding appreciably, but it reduced the catalytic activity of the holoenzyme nearly 5-fold. Removal of an additional single residue (i.e. removal of a total of 13 residues) weakened intersubunit binding approximately 80-fold. Paradoxically, this partially restored catalytic activity to approximately 40% of that of the wild-type holoenzyme. None of these truncations materially affected the Km values for ribulose-1,5-bisphosphate or CO2. Removal of all 20 residues of the irregular arm (thereby deleting the conserved region of contact with large subunits) totally abolished the small subunit's ability to bind to large subunits to form a stable holoenzyme. However, this truncated small subunit was still synthesized by the E. coli cells. These data are interpreted in terms of the role of the amino-terminal arm of the small subunit in maintaining the structure of the holoenzyme.     

Killer whale (Orcinus orca) reproduction at Sea World

Sea World has maintained killer whales (Orcinus orca) since 1965. The total killer whale inventory (1965–1993) has included 39 whales (25 females, 14 males); 28 were wild-caught and 11 captive-born, including one second-generation calf. As of September, 1993, there were 19 whales in the breeding program. Ten of these whales (53%) were captive-born, either at Sea World or other facilities in North America. The live wild-caught whales ranged in estimated age from 12–27 years (x? ± sd = 17.6 ± 4.2 years). The captive-born whales ranged in age from <1 to 8 years. In the Sea World breeding program (through September, 1993), there have been nine live births and one stillbirth, with eight calves part of the current inventory. Births occurred from July to February. Calving intervals ranged from 32–58 months. Female age at birth of first calves ranged from 8 years to an estimated 17 years (x? ± sd = 12.7 ± 3.0 years). Gestation, based on conception estimates from serum progesterone analysis, averaged 17 months (x? ± sd = 517 ± 20 days), but successful pregnancies with viable calves occurred from 15–18 months (468–539 days). Females, in the presence and absence of males, were polyestrus with periods of cycling interspersed with individually variable noncycling (presumed anestrous) periods ranging from 3–16 months. Mean serum progesterone levels (±se) were as follows: noncycling periods = 121 ± 20 pg/ml; peak elevations during nonconceptive ovulatory (estrous) cycles = 3,962 ± 2,280 pg/ml; first pregnancies = 14,592 ± 3,854 pg/ml; second pregnancies = 8,389 ± 395 pg/ml; and third pregnancy = 8,180 ± 4,556. © 1995 Wiley-Liss, Inc.     

Signaling and host cell invasion by Trypanosoma cruzi

Signal transduction events triggered in mammalian host cells by the obligate intracellular parasite Trypanosoma cruzi are required for invasion. Infective T. cruzi trypomastigotes elicit Ca2+ signaling in mammalian host cells and activate transforming growth factor-beta receptor signaling pathways. The elevation of Ca2+ in T. cruzi, induced by host-cell contact, is also required for invasion, extending the concept of host-pathogen 'cross-talk' to invasive protozoan pathogens.