Altered immunoreactivity of complexin protein in prefrontal cortex in severe mental illness.

Recent imaging and postmortem studies suggest that impaired connectivity is involved in the pathophysiology of schizophrenia and major affective disorders. We investigated the presynaptic proteins complexin (Cx) I and Cx II in postmortem prefrontal cortex in schizophrenia (n = 13; six suicide, seven nonsuicide), major depression (n= 11, all suicide) and controls (n = 11) with an enzyme-linked immunoadsorbent assay (ELISA). Overall analysis indicated a significant difference between groups (F = 3.93, P = 0.007). Cx I (enriched in inhibitory terminals) was decreased 33% in schizophrenia (26% in schizophrenia/nonsuicide, 42% in schizophrenia/suicide) and 27% in major depression. Cx II (enriched in excitatory terminals) was not significantly different. Analysis of the ratio of Cx II/Cx I was carried out as an indication of the balance of excitatory to inhibitory terminals. A significant difference between groups (ANOVA, F = 6.42, P = 0.005) was observed. The mean value of Cx II/Cx I was significantly increased by 34% in schizophrenia (26% in schizophrenia/nonsuicide and 43% in schizophrenia/suicide) and by 32% in depression compared with control (Student-Newman-Keuls test, P = 0.05). Immunoreactivities of the two complexins were highly correlated in all groups. However, compared with controls and depression, samples from cases with schizophrenia appeared to have relatively less Cx I for similar amounts of Cx II. Immunocytochemical studies of rat frontal cortex after 3 weeks treatment with chlorpromazine, trifluoperazine or haloperidol revealed no differences in complexins, synaptophysin, SNAP-25, syntaxin or VAMP in comparison with animals treated with vehicle. Alterations of complexins may contribute to the molecular substrate for abnormalities of neural connectivity in severe mental disorders.

Detergent inhibits 70-90% of responses to intravenous endotoxin in awake sheep.

Sheep have reactive pulmonary intravascular macrophages, which are essential for the marked pulmonary vascular response to infusions of small quantities of endotoxin. In another species with reactive pulmonary intravascular macrophages, horses, our laboratory found that an intravenous biosafe detergent, tyloxapol, inhibited some systemic and pulmonary responses to endotoxin (Longworth KE, Smith BL, Staub NC, Steffey EP, and Serikov V. Am J Vet Res 57: 1063-1066, 1996). We determined whether the same detergent would inhibit endotoxin responses in awake sheep. In 10 awake, instrumented sheep with chronic lung lymph fistulas, we did a control experiment by intravenously infusing 1 microg/kg Escherichia coli endotoxin. One week later, we gave 40 micromol/kg tyloxapol intravenously 1-4 h before infusing the same dose of endotoxin. In these paired studies, we compared pulmonary hemodynamics, lung lymph dynamics, body temperature, circulating leukocyte concentrations, and circulating tumor necrosis factor for 6 h. In all 10 sheep, tyloxapol blocked 80-90% of the pulmonary responses and 70-90% of the systemic responses. Tyloxapol is safe, inexpensive, easy to use, and effective immediately. It may be a clinically useful approach to contravening many of the effects of endotoxemia, in humans as well as animals.

The comparative biology of pulmonary intravascular macrophages.

Pulmonary intravascular macrophages are an important part of the mononuclear phagocyte system in some species of mammals, mainly sheep and other ruminants, pigs, and horses. These cells phagocytize foreign particles, cell debris and pathogens that pass through the pulmonary circulation. Species with intravascular macrophages localize intravenously injected tracer particles and bacteria predominantly in the lung rather than the liver, and exhibit pulmonary hypertension when these cells are activated. Both in vivo and in vitro studies show that pulmonary intravascular macrophages have distinct secretory and immune capabilities. Consequently, the pulmonary intravascular macrophages play an important role in pulmonary inflammation in species that have them

Effects of road transport on indices of stress in horses.

Stress associated with road transport is believed to be a significant contributor to the pathogenesis of post transport respiratory disease in horses. To determine the effects of road transport on pulmonary function, pulmonary aerosol clearance rates were measured in 4 horses 24 h before, and immediately after, 24 h of road transport by delivering aerosolised 99mtechnetium-labelled diethylenetriaminepentacetate (99mTc-DTPA) to the lungs and monitoring its washout. Each horse was transported twice, once while the trailer was equipped with a leaf-spring suspension and bias-ply tyres (trailer's original equipment, smooth ride) and once while the trailer was equipped with a torsion-bar suspension and normal pressure radial tyres (rough ride) in order to generate different ride characteristics. Before transport, blood was drawn from each horse for haematology and measurement of serum cortisol concentration; 24 h rates of hay and water intake and faecal output were recorded for each horse. Horses were then transported, 2 at a time, over a 128 km circular route of predominantly rural freeways at a constant speed of 72 km/h for 24 h. Horses were rested by stopping the trailer every 3.75 h for 0.25 h. During transport, heart rates (continuous 1 min averages), rates of hay and water intake and rates of faecal output were measured. Ammonia (NH3) and carbon monoxide (CO) concentrations were measured within the trailer and temperatures (wet bulb [WB], dry bulb [DB] and black globe [BG]) within the trailer were recorded each minute. Immediately after each experiment blood was drawn for haematology and measurement of pulmonary aerosol clearance rates were measured. For control studies, horses were housed in their stalls while heart rates were measured for 24 h. Slopes calculated from the 99mTc-DTPA clearance curves for pretransport horses were not significantly different from post transport clearance slopes. Pretransport mean 99mTc-DTPA clearance half-lives (T50, left lung mean +/- s.d. 41.7 +/- 15.8 min, right lung 44.6 +/- 19.1 min) were not significantly different from post transport T50 (left lung 53.5 +/- 14.0 min, right lung 52.0 +/- 11.6 min). Heart rates during transport were not affected by suspension type or trip order (the horse's first or second transport experiment) and were not significantly different from stall controls after the first 120 min of the experiment. Horses had increased red blood cell count, packed cell volume, haemoglobin, plasma protein and cortisol concentrations, and decreased body weights immediately post transport, indicating slight dehydration. Water and hay intake rates were significantly lower during transport than pretransport. Temperatures within the trailer were highest in the midafternoon and lowest in the early morning hours, but all temperatures measured in the trailer were within the comfort zone for large homeotherms. Ammonia and CO concentrations in the trailer during the transport period were within acceptable limits for human exposure. However, respirable articulates in the atmosphere were elevated above safe concentrations for human exposure.

Ultrastructural quantification of pulmonary intravascular macrophages in newborn and 2-week-old lambs.

BACKGROUND: Pulmonary intravascular macrophages are resident cells in the pulmonary circulation of sheep. Sheep, unlike species without pulmonary intravascular macrophages, exhibit pulmonary hypertension in response to intravenously injected particles. We reported in lambs that pulmonary vascular reactivity to intravenous particles increases with age as the population of intravascular macrophages develops. Preliminary quantitative histologic data showed that newborn lambs are born with few intravascular macrophages, but a large population develops over 2 weeks after birth. In this study, we present a complete quantitative analysis at the ultrastructural level. METHODS: We fixed five newborn and five 2-week-old lamb lungs by vascular perfusion and examined the tissue by electron microscopy. RESULTS: The fraction of capillary lumen taken up by intravascular macrophages/ monocytes is about three times greater in the lungs of 2-week-old lambs than that in newborn lambs (16% vs. 5%; P < 0.05). The fraction of capillary surface density associated with intravascular macrophages/monocytes is about three times greater in 2-week-old lambs than that in newborn lambs (8% vs. 3%; P < 0.05). The number of macrophages more than doubles with age (16 +/- 4 vs. 7 +/- 2; P < 0.05) and the estimated size (volume-weighted mean volume) increases by more than 1.5 times (294 +/- 46 microns 3 vs. 184 +/- 29 microns 3; P < 0.05). CONCLUSIONS: These data agree closely with Monastral blue retention by the lung (reported previously); there are more than twice as many mature pulmonary intravascular macrophages at 2 weeks than at 1 day after birth, and the cells are 1.5 times larger.

Use of detergent to prevent initial responses to endotoxin in horses.

OBJECTIVE: To determine whether a detergent can prevent most of the early effects of i.v. infusion with Escherichia coli endotoxin (< 100 ng/kg of body weight) in horses: marked pulmonary hypertension, acute leukopenia, and fever. ANIMALS: 8 healthy adult horses (4 male, 4 female), 415 to 615 kg. DESIGN AND PROCEDURE: Control and detergent experiments were performed in each horse while it was awake but sedated. In control experiments, 10 to 100 ng of E coli endotoxin/kg was given. In detergent experiments, 100 mg of detergent/kg was given 1 hour before injecting endotoxin, similar to the control experiments. RESULTS: In control experiments, pulmonary arterial pressure increased transiently over 40 minutes by 33 +/- 8 mm of Hg (mean +/- SD; P < 0.001), then returned to baseline. Circulating leukocytes decreased to 47 +/- 19% (P < 0.02) of baseline by 1 hour after endotoxin, then increased above baseline by 6 hours. Rectal temperature increased by 0.7 +/- 0.4 C (P < 0.01). In detergent experiments, the increase in pulmonary arterial pressure was much less than that in the control experiments (8 +/- 7 mm of Hg; P < 0.001). Circulating leukocytes did not decrease, and the increase in rectal temperature after endotoxic was blocked. CONCLUSIONS: This attenuation of te response to endotoxin may occur because the normal steps in the response of pulmonary intravascular macrophages (ie, endocytosis of endotoxin and subsequent release of inflammatory mediators) are altered by the detergent. This low-technology, inexpensive, and safe treatment could be an important new clinical tool for veterinarians in combating endotoxemia.

Structural determinants of maximal O2 transport in muscles of exercising foxes.

The arctic blue fox (Alopex lagopus) has a specific maximal oxygen consumption (VO2 max/Mb = 3.6 ml O2.s-1.kg-1) that is approximately 1.6-fold greater than those of dogs and horses. The fox has one of the highest body mass specific skeletal muscle mitochondrial volumes (V(mt,m)/Mb = 44 cm3.kg-1) among mammalian athletic species matching its higher VO2 max/Mb. The structural components related to capillaries, such as specific capillary length density (J(c)/Mb = 348 km.kg-1) and specific capillary volume (V(c)/Mb = 4.8 ml.kg-1), are not greater in the fox than in the larger athletes. Because a greater specific muscle diffusing capacity for oxygen (DTO2/Mb) is not utilized by the fox to achieve a higher VO2 max/Mb, a higher pressure difference for diffusion in the muscle capillaries is the alternative explanation for the fox's higher VO2 max/Mb. This mechanism is suggested by the fox's higher arterial and mixed venous capillary PO2 (120 mm Hg and 37 mm Hg, respectively) and its shorter mean muscle capillary transit time for blood (tc = 0.28 sec) compared to larger species.

Capillary blood transit time in muscles in relation to body size and aerobic capacity.

The mean minimal transit time for blood in muscle capillaries (tc) was estimated in six species, spanning two orders of magnitude in body mass and aerobic capacity: horse, steer, dog, goat, fox and agouti. Arterial (CaO2) and mixed venous (CvO2) blood O2 concentrations, blood hemoglobin concentrations ([Hb]) and oxygen uptake rates were measured while the animals ran on a treadmill at a speed that elicited the maximal oxygen consumption rate (VO2max) from each animal. Blood flow to the muscles (Qm) was assumed to be 85% of cardiac output, which was calculated using the Fick relationship. Total muscle capillary blood volume (Vc) and total muscle mitochondrial volume were estimated by morphometry, using a whole-body muscle sampling scheme. The tc was computed as Vc/Qm. The tc was 0.3-0.5 s in the 4 kg foxes and agoutis, 0.7-0.8 s in the 25 kg dogs and goats, and 0.8-1.0 s in the 400 kg horses and steers. The tc was positively correlated with body mass and negatively correlated with transcapillary O2 release rate per unit capillary length. Mitochondrial content was positively correlated with VO2max and with the product of Qm and [Hb]. These data suggested that Qm, Vc, maximal hemoglobin flux, and consequently tc, are co-adjusted to result in muscle O2 supply conditions that are matched to the O2 demands of the muscles at VO2max.

Pulmonary intravascular macrophages in horses and ponies.

Seven horses (4 anesthetized and 3 awake) and 2 ponies (anesthetized) were studied to evaluate the high sensitivity of the pulmonary circulation of the horse to various blood-borne particles, and to establish the presence of intravascular macrophages in the lung. Pulmonary and systemic pressures and cardiac output before and during particle injection were measured in some animals. An anesthetized foal had a large increase in pulmonary arterial pressure (32 and 34 mm of Hg) within 1 minute of IV administration of small test doses of radioactively labeled liposomes (2.5 mumol/kg of body weight) or a 1% suspension of blue pigment (0.3 ml/kg), respectively. Quantitative real-time gamma camera imaging of the foal revealed high retention of the labeled liposomes during the first pass through the lungs; retention persisted throughout the experiment. Postmortem analysis revealed 55 and 47% lung retention of liposomes and blue pigment, respectively. The 2 anesthetized ponies had increased pulmonary artery pressure of 34 +/- 7 mm of Hg, decreased cardiac output, and 42% lung retention after administration of 1% blue pigment (0.2 ml/kg), whereas 3 awake horses had increased pressure of 28 +/- 9 mm of Hg after 1.8 x 10(8) (1.8-microns-diameter) latex microspheres/kg. None of the injected particles caused vascular obstruction, and they do not cause pulmonary vascular reactivity in species that lack pulmonary intravascular macrophages. Finally, 3 horses (1 anesthetized and 2 awake) were infused IV with small doses of the blue pigment, and their lungs were perfusion-fixed to identify specific labeling of the pulmonary intravascular macrophages.(ABSTRACT TRUNCATED AT 250 WORDS)

Development of pulmonary intravascular macrophage function in newborn lambs.

We sought to determine whether pulmonary intravascular macrophages are involved in pulmonary vascular sensitivity to intravenously injected particles in sheep. We estimated that newborn lambs have few of these macrophages at birth but develop a 10-fold greater density within 2 wk. Awake, chronically instrumented newborn lambs showed no change in pulmonary vascular driving pressure (pulmonary arterial minus left atrial pressure) after injection of either liposomes [2 +/- 3 (SD) cmH2O; n = 5] or Monastral blue particles (3 +/- 2 cmH2O; n = 6) and showed no net pulmonary production of thromboxane B2, the stable metabolite of the vasoconstrictor thromboxane A2. In contrast, five of those lambs 2 wk later showed both an increase in pulmonary vascular driving pressure after injection of liposomes and Monastral blue (20 +/- 16 and 25 +/- 15 cmH2O, respectively; P < 0.05) and net pulmonary production of thromboxane B2 (171 +/- 103 and 429 +/- 419 pg/ml plasma, respectively; P < 0.05). Older lambs (n = 5) had higher pulmonary uptakes than newborn lambs (n = 6) of radioactive liposomes (47 +/- 13 vs. 12 +/- 10%; P < 0.01) and Monastral blue (53 +/- 6 vs. 21 +/- 10%; P < 0.05). We conclude that pulmonary intravascular macrophages are responsible for the sensitivity of sheep to intravenous foreign particles and are essential for a cascade of processes leading to microvascular injury.

O2 delivery at VO2max and oxidative capacity in muscles of standardbred horses.

The purpose of this study was to describe the relationships between 16 physiological, biochemical, and morphological variables presumed to relate to the oxidative capacity in quadriceps muscles or muscle parts in Standardbred horses. The variables included O2 delivery (blood flow) and mean capillary transit time (MTT) during treadmill locomotion at whole animal maximal O2 consumption (VO2max, 134 +/- 2 ml.min-1 x kg-1), capillary density and capillary-to-fiber ratio, myoglobin concentration, oxidative enzyme activities, glycolytic enzyme activities, fiber type populations, and fiber size. These components of muscle metabolic capacity were found to be interrelated to varying degrees using correlation matrix analysis, with lactate dehydrogenase activity showing the most significant correlations (n = 14) with other variables. Most of the "oxidative" variables occurred in the highest quantities in the deepest muscle of the group (vastus intermedius) and in the deepest parts of the other quadriceps muscles where the highest proportions of type I fibers were localized. The highest blood flow measured with microspheres in the muscle group during exercise was in vastus intermedius muscle (145 ml.min-1 x 100 g-1), and the lowest was in the superficial part of rectus femoris muscle (32 ml.min-1 x 100 g-1). Average muscle blood flow during exercise at whole animal VO2max was 116 ml.min-1 x 100 g-1. Because skeletal muscle comprised 43% of total body mass (453 +/- 34 kg), total muscle blood flow was estimated at 226 l/min, which was approximately 78% of total cardiac output (288 l/min).(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of interstitial edema on lung lymph flow in goats in the absence of filtration.

We tested the effect of interstitial edema on lung lymph flow when no filtration occurred. In 16 anesthetized open-thorax ventilated supine goats, we set pulmonary arterial and left atrial pressures to nearly zero and measured lymph flow for 3 h from six lungs without edema and ten with edema. Lymph flow decreased exponentially in all experiments as soon as filtration ceased. In the normal lungs the mean half time of the lymph flow decrease was 12.7 +/- 4.8 (SD) min, which was significantly shorter (P less than 0.05) than the 29.1 +/- 14.8 min half time in the edematous lungs. When ventilation was stopped, lymph flow in the edematous lungs decreased as rapidly as in the normal lungs. The total quantity of lymph after filtration ceased was 2.7 +/- 0.8 ml in normal lungs and 9.5 +/- 6.3 ml in edematous lungs, even though extravascular lung water was doubled in the latter (8.4 +/- 2.4 vs. 3.3 +/- 0.4 g/g dry lung, P less than 0.01). Thus the maximum possible clearance of the interstitial edema liquid by the lymphatics was 6.3 +/- 4.8%. When we restarted pulmonary blood flow after 1-2 h in four additional goats, lymph flow recovered within 30 min to the baseline level. These findings support the hypothesis that lung lymph flow originates mainly from alveolar wall perimicrovascular interstitial liquid and that the contribution of the lung lymphatic system to the clearance of interstitial edema (bronchovascular cuffs, interlobular septa) is small.

High rate of O2 consumption in exercising foxes: large PO2 difference drives diffusion across the lung.

The fox has one of the highest mass specific rates of maximal oxygen consumption (VO2max/Mb) that has been measured, yet its specific pulmonary diffusing capacity (DLO2/Mb, measured morphometrically) is similar to that of most mammalian species. It achieves a high O2 flux per unit DLO2 with a large partial pressure difference driving O2 diffusion from alveolar gas to capillary blood (PAO2-PbO2). This paper explores the mechanisms that the fox utilizes to achieve this large pressure difference and the extent to which it exploits its structural diffusing capacity. Foxes were exercised on a treadmill at maximal rates of O2 uptake. The following parameters were measured or calculated: arterial and mixed venous PO2, PCO2, pH and O2 concentration of the blood, cardiac output, hemoglobin concentration and O2 equilibrium curve of the blood, and morphometric estimates of pulmonary capillary volume and pulmonary diffusing capacity for O2. These data were used to calculate pulmonary capillary transit time and the time course of the change in O2 concentration and PO2 of the blood as it transits the lung. The fox has a morphometric pulmonary diffusing capacity of 0.098 ml O2.sec-1.mm Hg-1.kg-1. At VO2 max (3.6 ml O2.sec-1.kg-1) the fox hyperventilates, resulting in a high PAO2 (124 mm Hg); it also maintains a low PbO2 (88 mm Hg) by having a short transit time (0.13 sec) due to a high specific cardiac output (25 ml.sec-1.kg-1). Our calculations indicate that at VO2max the fox uses almost all of the pulmonary capillary transit time for O2 equilibration, in contrast to other species.

Oxygen transport during exercise in large mammals. I. Adaptive variation in oxygen demand.

This study investigated mechanisms used by horses and steers to increase O2 uptake and delivery (VO2) from resting to maximal rates and identified the mechanisms that enable horses to achieve higher maximal rates of O2 consumption (VO2max) than steers. VO2 and circulatory variables were measured while Standardbred trotting horses and steers (450-kg body mass) stood quietly and ran on a treadmill at speeds up to those eliciting VO2max. As VO2 increased in both species, heart rate and circulating hemoglobin (Hb) concentration increased, thereby increasing O2 delivery by the circulation, while cardiac stroke volume remained unchanged. At VO2max arterial PCO2 increased from its resting value in horses but was unchanged in steers, and arterial PO2 decreased in both species. Although the horses hypoventilated and were hypoxemic at VO2max, no significant decrease in arterial Hb saturation occurred. VO2max of the horses was 2.6 times higher than that of the steers and was associated with a 100% larger cardiac output, 100% larger stroke volume, and 40% higher Hb concentration, whereas heart rates at VO2max were identical in the two species. The higher cardiac output of the horses at VO2max resulted from a 1.2-fold higher mean arterial pressure and 1.6-fold lower peripheral tissue resistance (associated with a larger skeletal muscle capillary bed). Both the magnitude of the difference in VO2max between horses and steers and the mechanisms used to achieve it are the same as observed in smaller pairs of mammalian species with large variation in aerobic capacity.

Blood gas measurements during exercise: errors due to temperature correction.

This study assessed the degree to which correcting blood gas measurements to rectal temperature (Tre) rather than to the temperatures at which gas exchange occurs [pulmonary arterial (Tpa) or intramuscular (Tm)] introduces errors into blood gas analysis of exercising mammals. Horses and steers weighing 450 kg were run on a treadmill at speeds up to those eliciting maximal rates of O2 consumption (VO2max), and temperatures were measured in various body compartments. In both species Tpa rose faster than Tre during the run, the degree of dissociation being a function of exercise intensity and duration. Tm was measured only in horses, and it rose faster than Tpa during the run and decreased more slowly postrun. Correcting blood gas values measured at an analyzer temperature of 37 degrees C to Tre without accounting for transient increases during the run of Tpa and Tm that were never reflected in Tre significantly biased estimates of blood gases. The biased estimates erroneously indicated that both species experienced more severe hypoxemia than they actually did at VO2max and masked the hypercapnia experienced by the horses at VO2max.