Spike firing allometry in avian intrapulmonary chemoreceptors: matching neural code to body size.

Biological rates in small animals are usually higher than those in large animals, yet the maximal rate of action potential (spike) generation in sensory neurons encoding rate functions is similar in all animals, due to the conserved genetics of voltage-gated ion channels. Therefore, sensory signals that vary at rates approaching maximal spike generation rate, as might occur in animals of diminished body size, may require specialized spike coding to convey this information. To test whether spike coding scales allometrically in sensory neurons monitoring signals that change frequency with body size, we recorded action potentials from 70 avian intrapulmonary chemoreceptors (IPC), respiratory neurons that detect lung CO2 changes during breathing, in five different avian species ranging in size from body mass Mb=0.045 kg (lovebirds) to 5.23 kg (geese). Since breathing frequency scales approximately to Mb-1/4 (higher in small birds, lower in large birds), we reasoned that IPC discharge frequencies may also scale to maintain spike information transmission within each breath. We found that phasic action potential discharge pattern, as quantified by the peak discharge rate and the magnitude of spike frequency adaptation, scaled between Mb-0.22 and Mb-0.26, like breathing rate (P<0.05). Previously published values of peak discharge rate in IPC also fit this allometric relationship. We suggest that mass-dependent scaling of neural coding may be necessary for preserving information transmission with decreasing body size.

DNA from uncultured organisms as a source of 2,5-diketo-D-gluconic acid reductases.

Total DNA of a population of uncultured organisms was extracted from soil samples, and by using PCR methods, the genes encoding two different 2,5-diketo-D-gluconic acid reductases (DKGRs) were recovered. Degenerate PCR primers based on published sequence information gave internal gene fragments homologous to known DKGRs. Nested primers specific for the internal fragments were combined with random primers to amplify flanking gene fragments from the environmental DNA, and two hypothetical full-length genes were predicted from the combined sequences. Based on these predictions, specific primers were used to amplify the two complete genes in single PCRs. These genes were cloned and expressed in Escherichia coli. The purified gene products catalyzed the reduction of 2,5-diketo-D-gluconic acid to 2-keto-L-gulonic acid. Compared to previously described DKGRs isolated from Corynebacterium spp., these environmental reductases possessed some valuable properties. Both exhibited greater than 20-fold-higher kcat/Km values than those previously determined, primarily as a result of better binding of substrate. The Km values for the two new reductases were 57 and 67 microM, versus 2 and 13 mM for the Corynebacterium enzymes. Both environmental DKGRs accepted NADH as well as NADPH as a cosubstrate; other DKGRs and most related aldo-keto reductases use only NADPH. In addition, one of the new reductases was more thermostable than known DKGRs.

Effects of embryonic CO2 exposure on the adult ventilatory response in quail: does gender matter?

To test the hypothesis that the adult ventilatory response to CO2 can be modified by exposure to CO2 during development, we exposed Japanese quail (Coturnix japonica) from two populations to 2% CO2 throughout embryonic development. The ventilatory response to 6% CO2 was subsequently measured in control and CO2-exposed quail following a 6-9 week deacclimation period. In one population of quail, CO2-exposed female, but not male, quail had a reduced ventilatory response to 6% CO2 as adults (P<0.001). Although control and CO2-exposed quail had similar ventilation while breathing air, CO2-exposed females exhibited a 26% reduction in mean ventilation at 6% CO2, primarily due to a reduced tidal volume. In contrast, the identical incubation treatment had no effect on the ventilation of either gender in a second population of quail. It appears that developmental plasticity in the hypercapnic ventilatory response may generally be greater in females, although this conclusion may vary depending on genetic factors.

An improved automated immunoassay for C-reactive protein on the Dimension clinical chemistry system.

Recent clinical data indicate that the measurement of the concentration of C-reactive protein (CRP) requires a higher sensitivity and wider dynamic range than most of the current methods can offer. Our goal was to develop a totally automated and highly sensitive CRP assay with an extended range on the Dimension((R)) clinical chemistry system based on particle-enhanced turbidimetric-immunoassay (PETIA) technology. The improved method was optimized and compared to the Binding Site's radial immunodiffusion assay using disease state specimens to minimize interference. Assay performance was assessed on the Dimension((R)) system in a 12-instrument inter-laboratory comparison study. A split-sample comparison (n = 622) was performed between the improved CRP method on the Dimension system and the N Latex CRP mono method on the Behring Nephelometer, using a number of reagent and calibrator lots on multiple instruments. The method was also referenced to the standard material, CRM470, provided by the International Federation of Clinical Chemistry (IFCC). The improved CRP method was linear to 265.1 mg/l with a detection limit between 0.2 and 0.5mg/l. The method detects antigen excess from the upper assay limit to 2000 mg/l, thereby allowing users to retest the sample with dilution. Calibration was stable for 60 days. The within-run reproducibility (CV) was less than 5.1% and total reproducibility ranged from 1.1 to 6.7% between 3.3 and 265.4 mg/l CRP. Linear regression analysis of the results on the improved Dimension method (DM) versus the Behring Nephelometer (BN) yielded the following equation: DM = 0.99 x BN - 0.37; r = 0.992. Minimal interference was observed from sera of patients with elevated IgM, IgG and IgA. The recovery of the IFCC standard was within 100 +/- 7 % across multiple lots of reagent and calibrator. The improved CRP method provided a sensitive, accurate and rapid approach to quantify CRP in serum and plasma on the Dimension clinical chemistry system. The ability to detect antigen excess eliminated reporting falsely low results caused by the 'prozone effect'.

Automated homogeneous immunoassay for gentamicin on the dimension clinical chemistry system.

BACKGROUND: Monitoring of the concentration of gentamicin in serum and plasma during therapy is widely recommended and practiced in hospitals. Our aim was to develop a homogeneous immunoassay based on particle-enhanced turbidimetric inhibition immunoassay technology to quantify gentamicin on the Dimension clinical chemistry system. METHODS: Assay performance was assessed on each of the Dimension models in a 15-instrument interlaboratory comparison study. A split-sample comparison (n = 1171) was also performed between the gentamicin methods on the Dimension system and the Abbott TDx analyzer, using multiple reagent and calibrator lots on multiple instruments. RESULTS: The Dimension method was linear to 25.1 micromol/L (12.0 microg/mL) with a detection limit of 0.63 micromol/L (0.3 microg/mL). Calibration was stable for 30 days. The within-run imprecision (CV) was <1.3%, and total imprecision ranged from 1.8% to 3.2% between 4.2 micromol/L (2.0 microg/mL) and 16.7 micromol/L (8.0 microg/mL) gentamicin. Linear regression analysis of the results on the Dimension method (DM) vs the Abbott TDx yielded the following equation: DM = 0.98TDx - 0.42; r = 0.987. Minimal interference was observed from structurally related compounds such as sagamicin, netilmicin, and sisomicin. CONCLUSION: The monoclonal antibody used in this method has similar reactivities toward the individual gentamicin subspecies C1, C1a, and C2, thus providing analytical recovery not significantly dependent on relative subspecies concentrations.

In vivo blood oxygen binding in hypoxic lesser spear-nosed bats: relationship to control of breathing.

The hypoxic ventilatory threshold of many mammals correlates with their hemoglobin-oxygen affinity (P50). Yet, in some small mammals ventilation actually declines, rather than increases, with exposure to decreasing PaO2; their air convection requirement (V(E)/V(O2)), however, is elevated in hypoxia. We propose that the threshold of the hypoxic V(E)/V(O2) of small mammals coincides with the inflection ('knee') of their in vivo O2 equilibrium curve (O2EC). In vivo blood gas and pH data were obtained from normoxic and hypoxic lesser-spear nosed bats, Phyllostomus discolor; in vitro blood O2EC were also generated for normoxic bats at 32 and 37 degrees C and at three P(CO2)'s. The hypoxic V(E)/V(O2) threshold of P. discolor occurs at PaO2 = 39 Torr; the corresponding in vivo O2 saturation is 0.70, approximating the inflection of the O2EC. This animal has a high blood O2 affinity (P50 = 27.5 Torr at pH 7.40 and 37 degrees C; P50 = 30.8 Torr at in vivo pH of 7.31 and TB of 37.4 degrees C). As PaO2 is reduced, a pronounced hypoxia-induced respiratory alkalosis and hypothermia help maintain SaO2 near the O2EC inflection (0.64-0.70 S(O2)).

Ventilatory, cardiovascular and metabolic responses to hypoxia and hypercapnia in the armadillo.

Armadillos have a low resting metabolic rate and high hemoglobin affinity for their size, a rigid carapace and a semi-fossorial life style. These characteristics could contribute to unusual respiratory responses to hypoxia and hypercapnia which were investigated in this study. Ventilatory and oxygen consumption responses of six adult unanesthetized armadillos to 15, 12, 10 and 8% O2 and 1.5, 3, 5 and 7% CO2 were measured by barometric plethysmography and flow-through respirometry. A significant increase in ventilation occurred in response to 10 and 8% O2 but a decline in oxygen consumption only occurred at 8% inspired O2. The convection requirement response has a threshold at a PaO2 of approximately = 28 Torr which corresponds to a Hb saturation of approximately 70%. Ventilation increased in response to 3% and higher levels of CO2, with no change in oxygen consumption. The magnitude of the ventilatory response to CO2 was similar to other semi-fossorial mammals and less than that of nonburrowing species. However, the pattern of the response was unique in being largely a frequency response with little change in tidal volume, contrary to the tidal volume dominated response to hypercapnia typical of mammals. This feature, not shared by another Xenarthran, the sloth, who lacks a carapace, is likely attributable to the low respiratory system compliance and increased airway resistance resulting from the rigid carapace and small lungs of armadillos and emphasizes the importance of respiratory mechanics in determining breathing pattern.

How stiff is the armadillo? A comparison with the allometrics of mammalian respiratory mechanics.

Static respiratory mechanics were examined in the armadillo (Dasypus novemcinctus) and compared with allometric relationships newly derived for adult mammals from values in the literature. Normalised by body weight, chest wall compliance (Cw) in the armadillo is lower than predicted. Lung compliance (Cl) is also low in the armadillo, however it is appropriately matched to the resting lung volume (Vr) (ie. Cl/Vr infinity Mass0.0) and the ratio of Cw/Cl is appropriate for the size of the animal. Respiratory system resistance is high in the armadillo, presumably because of smaller airways associated with the small lung. The power of breathing in the armadillo is comparatively high, mainly due to the high resistive forces. Indeed, the oxidative cost of breathing is approximately double that of a mammal with similar Vr. Hypoxia or hypercapnia are known to invoke an attenuated ventilatory response in the armadillo and one that relies more on changes in frequency rather than volume. While such a breathing pattern helps to reduce the power of breathing it also compromises the degree of hyperventilation achieved.

Coordination of respiratory cycles with wingbeat cycles in the black-billed magpie (Pica pica)

Magpies fly with a variable pattern of wingstroke, including high-amplitude rapid flaps and low-amplitude slower flaps with interspersed brief glides. This allowed us to test the hypothesis that if phasic coordination between respiratory and wingbeat cycles is important mechanically and energetically, then, as a bird changes its wingbeat cycle, its respiratory cycle should change with it. We also tested the strength of the drive to coordinate respiratory to locomotor cycles by stimulating breathing with 5 % CO2 during flight. We found that magpies (N=5) do shorten their breath cycle time when they shorten their wingbeat cycle time and prolong their breath cycle time when they glide. When the coordination ratio of wingbeat cycles to breaths is 3:1, the pattern of phasic coordination ensures two upstrokes per inspiration and two downstrokes per expiration. Upstroke tends to coincide with the transition into inspiration or with early inspiration and late inspiration. Downstroke tends to coincide with the transition into expiration or with early expiration and late expiration. When magpies switch from a 3:1 ratio to a 2:1 ratio of wingbeat cycles to breaths, they shorten inspiratory time to ensure that upstroke occurs through most of inspiration and downstroke corresponds to the transition into expiration. These phasic coordination patterns ensure that the compression of the airsacs during downstroke can provide a net assistance to expiration and that the expansion of the airsacs with upstroke can provide a net assistance to inspiration. The failure of an atmosphere containing 5 % CO2 to disrupt these phasic coordination patterns between respiratory and locomotory cycles suggests that there may be a potent mechanical and energetic benefit to such coordination.

Ventilatory and metabolic responses of a bat, Phyllostomus discolor, to hypoxia and CO2: implications for the allometry of respiratory control.

The ventilatory and metabolic responses of lesser spear-nosed bats to hypoxia and hypercapnia were measured to determine whether these corresponded to preliminary allometries and a positive relationship between hypoxic ventilatory threshold and P50. Ventilatory responses of lesser spear-nosed bats to 3, 5 and 7% CO2 differed significantly from ventilation on air and each other. The magnitude of their ventilatory response to CO2 is consistent with the prediction of a smaller ventilatory response to hypercapnia in small compared to large mammals [% delta V varies MB0.130; Williams et al. (1994)]. Among 12, 10 and 8% O2 treatments only the ventilatory response to 8% O2 differed significantly from ventilation on air or the other treatments. Metabolic rate was significantly reduced at both 10 and 8% O2. The hypoxic ventilatory response of these bats does not support the prediction of a greater response in small compared to large mammals [% delta V veries MB0.273; Boggs and Tenney (1984)]. Their metabolic response is consistent with the hypoxic hypometabolism typical of small mammals, though not of comparable magnitude. The response, expressed as percent change in convection requirement (V/VO2), is also less than that observed in other small mammals. This relative insensitivity to hypoxia may be associated with this bat's unusually high affinity hemoglobin (P50 = 27.5 torr).

Scaling of hypercapnic ventilatory responsiveness in birds and mammals.

The possible relationship between CO2 responsiveness and body mass in birds was explored using newly acquired ventilatory data from the barn swallow, Hirundo rustica, and the pigeon, Columbia livia, and that from the literature on four other species. Ventilatory responsiveness (% delta V) of birds to 5% inspired CO2 is scaled to body mass to the 0.145 power (% delta V alpha Mb 0.145). A similar allometric relationship exists for data on 7 species of eutherian mammals taken from the literature (% delta V alpha Mb0.130). The The reduced responsiveness to CO2 in small birds and mammals may be related to an elevated hypoxic ventilatory sensitivity, as demonstrated in mammals (Boggs and Tenney, Respir. Physiol. 58: 245-251, 1984). These scaling relationships may reflect a mechanism for minimizing the inhibition of ventilation resulting from excessive loss of CO2 which thereby permits a higher hypoxic ventilatory response in small species. Other mechanisms, however, could include size related differences in mechanics or alveolar ventilation.

Ontogenetic modification of the hypercapnic ventilatory response in the zebra finch.

We investigated the effect of exposure to CO2 during development on the adult ventilatory response to CO2 of zebra finches. Developing zebra finches were exposed to chronic hypercapnic respiratory environments as embryos, as nestlings or during both developmental periods. Their responsiveness to CO2 was then measured following a 125-135 day post-fledging deacclimation period. At a FICO2 of 0.06, mean ventilation of adult finches in the three groups was 74.8, 61.3 and 65.8%, respectively, of that of finches reared under normocapnic conditions. These data indicate that adult acute ventilatory responsiveness to CO2 in birds may be in part determined by CO2 exposure during early development and may help explain the observation that fossorial and semifossorial species of birds and mammals, that naturally encounter high CO2 conditions in their burrows, have a blunted hypercapnic ventilatory response.

Chemosensitivity and breathing pattern regulation of the coatimundi and woodchuck.

An analysis of breathing pattern regulation was carried out on the coatimundi and woodchuck who represent two different volume-time patterns. It was found that the coati, with a short expiratory time as a fraction of total breath time, TE/TTOT, has a greater sensitivity to CO2 as represented by the slope and threshold of its ventilatory response. Breathing air the coati maintains post-inspiratory inspiratory activity (PIIA) of the posterior cricoarytenoid (PCA) through 51% of expiration, while the woodchuck, who is less sensitive to CO2 and has a long TE/TTOT, exhibits no PIIA of the PCA. The woodchuck also has a greater incidence and duration of end-expiratory pauses (or delayed inspiratory onset). The woodchuck does not demonstrate the usual inverse relationship between VT and TE in response to 5% CO2 and does not recruit PIIA of the PCA at this level of CO2. These data confirm the importance of CO2 chemosensitivity in regulation of TE. It is further demonstrated that interspecific differences in chemosensitivity among three mammals of the same size are reflected in regulation of TE but not in inspiratory 'drive' (as indicated by mean inspiratory flow, VT/TI).

The effect of diazepam enema on the rectal mucosa of rats.

The use of undiluted intravenous diazepam solution administered as an enema has been recommended for the acute management of seizures when intravenous access cannot be obtained. It is well absorbed, rapidly effective, and generally accepted as being safe when used in this manner. The effect of intravenous diazepam solution on the rectal mucosa has not been studied. This study demonstrates that there are no acute changes in the rectal mucosa of adult rats after administration of 10 to 20 times the recommended human dose.

Temperature regulation and basal metabolic rate in the spotted skunk, Spilogale putorius.

1. Metabolic rate, body temperature, and evaporative water loss of six spotted skunks were measured at air temperatures between 8 and 40 degrees C. 2. The mean metabolic rate of spotted skunks at thermoneutral air temperatures was 30.5% below that predicted by body mass. 3. Thermal conductance, body temperature, and rates of evaporative water loss were like those of similar sized mammals. 4. The non-elongate body form, omnivorous diet, and low level of activity of spotted skunks distinguish them from other mustelids and may account for their lower-than-expected basal metabolism.

Blood oxygen binding properties for the burrowing owl, Athene cunicularia.

Isocapnic O2 equilibrium curves (O2ECs) were generated for whole blood of 4 adult burrowing owls (Athene cunicularia) using thin film techniques. At in vivo pHa (7.49 +/- 0.02; mean +/- 1 SEM) and 41 degrees C, the PO2 at half saturation (P50) was 42.3 +/- 0.8 Torr. CO2 and fixed acid (H+) Bohr slopes (delta log P50/delta pH) were -0.46 +/- 0.01 and -0.42 +/- 0.02, respectively, demonstrating a small specific CO2 effect. CO2 and H+ Bohr slopes were saturation-independent between 0.1 and 0.9 S. Hill plots for Athene blood were non-linear; the Hill coefficient (n) increased from 2.6 below 0.4 S to 3.4 above 0.6 S. Owl equilibrium data were accurately described by the equation: S = [(7.7 x 10(6]/(P4 + 44P3 - 108P2 + 3.5 x 10(4)P) + 1]-1. This complex O2EC shape may result from Hb heterogeneity; isoelectric focusing showed 4 isoHbs with a molar ratio of 9:1:1:1. This study revealed no apparent adaptations of Athene blood for hypoxic and hypercapnic conditions. We conclude that the observed blood O2 binding properties promote tissue O2 delivery during periods of surface activity. While occupying its burrow, the owl compensates for moderate alterations in inspired gas composition partly through increased ventilation.

Effects of hypoxia and hypercapnia on VT, f, and VI of nestling and adult bank swallows.

The effects of hypoxia, hypercapnia, and hypoxic hypercapnia on ventilation, and breathing pattern in adult and nestling bank swallows (Riparia riparia) were assessed. The CO2 threshold above which inhaled minute volume (VI) increased significantly in adults and nestlings was 0.045. At each level of fractional concentration of inspired CO2 (FICO2), ventilation in nestlings was lower than that in adults. At a FICO2 of 0.09, VI of adults increased by 284%, whereas VI in nestlings changed 238%. Adult bank swallows also showed a blunted ventilatory response to hypoxia, and the nestling's response was similar to other birds. Adults exhibited greater changes in VI at all levels of hypoxic hypercapnia compared with nestlings. Combined hypoxic and hypercapnic stimuli had an additive effect on ventilation in both groups. Chronic exposure of nestlings to the hypercapnia and hypoxia within burrows seems to significantly alter their ventilatory response to these respiratory stimuli.

Differentiation of intramedullary neoplasms and cysts by MR.

To determine the MR criteria that are effective for differentiating intramedullary neoplasms from syringo- or hydromyelia, we reviewed MR scans made on prototype and commercial imagers of 33 patients with surgically confirmed cord abnormalities, including nine intramedullary neoplasms and 20 cysts (syringo- or hydromyelia). Two radiologists who did not know the clinical and radiologic diagnoses were asked to evaluate the scans with respect to cord expansion, distinctness of the disease margin, homogeneity, and signal intensity. These observations were correlated with the proved diagnoses. The combination of distinct margins and uniform signal intensity equal to that of CSF correlated consistently (88%) with spinal cord cysts. Other combinations were less reliable for diagnosing a cyst or tumor.