No abstract

Two nonaqueous fixatives, composed of fluorocarbon solvents with dissolved osmium tetroxide, were used to determine the feasibility of preserving the mucous coat in bovine and rat trachea for light and electron microscopy. Aqueous fixatives, while providing excellent cytological preservation, wash away the mucous lining, precluding ultrastructural analysis. Inclusion of ruthenium red or alcian blue within aqueous fixative improved retention of mucus, but provided incomplete, patchy results. Fixation with nonaqueous fluorocarbon solvent and dissolved osmium tetroxide preserved a continuous mucous epiphase layer above a clear hypophase layer. Subcomponents of the mucus included an electron dense surface layer, interrupted patches of mucus above the surface layer and electron dense membrane-like material within the mucus. This method of fixation will preserve mucus for light, scanning and transmission electron microscopy, using either intratracheal or immersion methods of fixation. The latter would enable use of materials from large animal models, autopsy or an abattoir.

Experimental conditions for measuring the effect of the noncaloric fat substitute olestra on the availability of dietary nutrients were established in the weanling domestic pig. To evaluate the tolerance of the pig for dietary fat levels similar to those in the human diet, groups were fed a standard corn-soy-based swine feed with and without 14% (30% of energy) added fat for 4 wk. To evaluate the adequacy of a purified diet to produce good growth, groups of pigs were fed purified diets providing 30% of energy from fat and micronutrients at 1, 1.3 or 1.6 times the NRC's requirements for 5- to 10-kg swine. Cumulative body weight gain, digestible feed efficiency and a lack of adverse effects showed that the pig can tolerate diets providing 30% of energy from fat and that a purified diet providing the NRC's requirements for micronutrients produces growth comparable to a nutritionally complete swine feed. To determine whether tissue concentrations of vitamins A, D, E and K in the pig respond to olestra and dietary concentrations of the vitamins, two groups were fed purified diet providing 1 or 1.6 times the NRC's requirements for micronutrients and 4.8% olestra. Significant increases occurred in the serum concentration of 25-hydroxyergocalciferol and liver concentrations of retinol and alpha-tocopherol with increasing dietary concentrations of the vitamins. Olestra reduced the tissue concentrations of vitamins A, D and E. Prothrombin time was not affected by dietary concentration of either phylloquinone or olestra. To determine the amount of UV light exposure required to produce 50-80% of vitamin D status from vitamin D3, a range typical of humans, two groups of pigs were fed the NRC requirement for vitamin D and exposed to 15 or 45 min/d of UV light. Serum concentration of 25-hydroxycholecalciferol increased with increased exposure time. UV exposure of 1-2 min/d was calculated to be sufficient to produce 50-80% of total vitamin D status from vitamin D3. No antemortem observations indicated an adverse olestra effect.

A B S T R A C T Once ventilatory acclinmatizationi beginsin sea level residents sojourning at high altitude, abrupt restoration of normal oxygen tensions will not restore ventilation to normal. We have investigated the role of cerebrospinal fluid (CSF) [He] in this sustained hyperventilation by measurinig CSF acid-base status in seven imien (lumbar) and five ponies (cisterinal) in normoxia, first at sea level anid then periodically over 13-24 h of"deacclimatization" after 3-5 d in hypoxia (PB = 440 mm Hg). After 1 h deaccliimcatization, hyperventilation continued at a level only slightly less than that obtained in chronic hypoxia (+ 1-2 mm Hg PA,O2), whereas CSF pH was either e(lual (in man) or alkaline (in pony, +0.02, P < 0.01) to sea level values. Between 1 and 12-13 h deacclimatization in all humans and ponies VA fell progressively (PA(,0( increased 4-7 mm Hg) and CSF pH became increasingly more acid (-0.02 to -0.05, P < 0.01). Between 12 acnd 24 h of normoxic deacclimatization in ponies, PAco, rose further toward norinal, coincideint with an increasing acidity in CSF (-0.02 pH). Similar negative correlations were found between changes in arterial pH and VA throughout normoxic deaccliimiatization. We conclude that [H+] in the lumbar or cisternal CSF is not the mediator of the continued hyperventilation and its gradual dissipation with time during normoxic deacclimatization from chronic hypoxia. These negative relationships of VA to CSF [H+] in normoxia are analogous to those previously shown during acclimatization to hypoxia.

Groups of weanling pigs (5 castrated males, 5 females per group) were fed purified diets containing the NRC's requirements for nutrients and 0, 1.1, 4.4 or 7.7% olestra for 12 wk. Graded concentrations of vitamins A, D2 and E were added at each olestra concentration. The primary purpose of the study was to establish relationships between dietary concentration of olestra and the amounts of vitamins A, D2 and E needed to restore tissue concentrations of these vitamins to control concentrations. A secondary purpose was to confirm that olestra does not affect the status of vitamin K or water-soluble nutrients. Liver concentrations of vitamins A, E and B12, iron and zinc and bone concentrations of ash, zinc, calcium and phosphorus, were measured in a group of pigs killed at the start of the study and in all pigs killed at wk 12. Growth, feed efficiency, hematology, clinical chemistry, blood concentrations of retinol, alpha-tocopherol, 25-hydroxyergocalciferol, 25-hydroxycholecalciferol, 1,25-dihydroxyvitamin D, folate, iron, total iron-binding capacity, zinc and calcium and adipose concentration of vitamin E were measured at 4-wk intervals. Prothrombin time was measured weekly for the control and 7.7% olestra groups, monthly for others. Relationships derived from measured tissue concentrations of vitamins A and E showed that constant amounts of the vitamins were required per unit mass of olestra in the diet to restore tissue concentrations to control values. Such a relationship could not be determined for vitamin D because exposure of the pigs to UV light resulted in an apparent interaction between vitamin D2 and vitamin D3. Olestra did not affect growth, digestible feed efficiency, vitamin K status or the status of the water-soluble micronutrients, in agreement with other studies in the pig.