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Biomedical subjects

R E Anderson

Publications and source records attributed to R E Anderson.

At least 307 records · Page 17Linked to original sources

Rabbit photoreceptor outer segments contain high levels of docosapentaenoic acid.

Docosapentaenoic acid (22:5 omega 6), a minor constituent (less than 4%) of photoreceptor outer segment membranes in all vertebrate species examined to date, comprises 23% of the fatty acids in total lipids from rabbit outer segment membranes. This fatty acid is a significant constituent of each of the three major phospholipid classes in these membranes. The levels of docosahexaenoic acid (22:6 omega 3), the major polyunsaturated fatty acid of most other vertebrate outer segments, was 20%. The sum of 22:5 omega 6 and 22:6 omega 3 in rabbit outer segment membrane lipids is similar to the amount of 22:6 omega 3 usually found in membranes from other vertebrate species.

Animals↗

Effects of inositol-1,4,5-trisphosphate injections into salamander rods.

Solitary rods were isolated by trituration of salamander (Ambystoma tigrinum) retinas. One barrel of an intracellular, double-barreled micropipette was used to record membrane voltage; the other barrel was used to pressure-inject inositol-1,4,5-trisphosphate. The injection of inositol-1,4,5 -trisphosphate induced a reversible hyperpolarization of the rod membrane. Injections of inositol-1,4,5-trisphosphate decreased the size of receptor potentials induced by dim lights. Conversely, light decreased the responses of the rod to injections of inositol-1,4,5-trisphosphate. These results suggest that inositol-1,4,5-trisphosphate might be involved in the modulation of rod membrane voltage during phototransduction.

Animals↗

Stimulation of carbonic anhydrase in osteoclasts by parathyroid hormone.

Changes in the acidity of osteoclasts were evaluated by direct measurement of the fluorescent intensity of osteoclasts exposed to acridine orange, a fluorescent weak base which becomes concentrated in acid-containing subcellular compartments. Parathyroid hormone (PTH) produced dose-dependent increases in fluorescent intensity; maximal increases in intensity occurred at doses between 3 and 10 micrograms PTH/ml of culture medium. Acetazolamide, a potent inhibitor of carbonic anhydrase, inhibited the increase in fluorescence induced by PTH, but this drug was less effective in reducing fluorescence in maximally than in submaximally stimulated osteoclasts, indicating that either more enzyme or more resistant enzyme was present in the PTH-stimulated cells. Because increased fluorescence of acridine orange is a sign of greater acidity, these results suggest that (1) PTH stimulates the acidity of osteoclasts, (2) carbonic anhydrase activity is necessary for maximum acidity, and (3) carbonic anhydrase is activated by PTH.

Acetazolamide↗

Fraction size, dose and time dependence of X ray induced late renal injury.

Histologic quantitation of renal radiation injury based on an intact mouse model correlates well with loss of renal mass, as measured by the ratio of right to left renal weight, radiation dose, and time after irradiation. Use of the radiation dose at which 50% of animals show renal tubular changes of a specified grade (ED50), allows comparison of 1, 5 and 15 fraction exposures. The X-ray dose necessary to effect significant injury (Grade 3 change) by six months after irradiation (ED50) was 11.6, 25.7, and 44.0 Gy for single, 5, and 15 fractions, respectively. Isoeffect plots of total radiation dose as a function of fraction number are linear on logarithmic coordinates, indicating that this relationship followed a power function. When plotted as inverse total dose versus dose per fraction, the six month post-irradiation data were linear, but the 12 month data were not. Lack of linearity is not in agreement with the multi-fraction isoeffect linear quadratic model.

Animals↗

Catabolism of myo-inositol to precursors utilized for de novo glycerolipid biosynthesis.

Systemic injection of [2-3H]myo-inositol into frogs resulted in the incorporation of more than half of the label into glycerolipid classes other than phosphoinositides in retinal rod outer segment membranes. Following methanolysis and differential extraction of isolated lipid classes, radioactivity was recovered primarily in the aqueous phase. After phospholipase C hydrolysis of the total membrane lipids, 97% of the radioactivity was extractable with organic solvents, and 70% of the label in lipids was in 1,2-diglycerides. These results indicate that the label was incorporated primarily into the glyceryl moiety of the membrane glycerolipids. Intraocular injection of frog eyes or in vitro incubation of frog retinas with [2-3H]myo-inositol resulted in the incorporation of radioactivity almost exclusively into phosphoinositides in rod outer segment membranes. Incubation of retinas with [U-14C]glucuronic acid did not result in the formation of labeled retinal lipids. These results suggest that myo-inositol can be catabolized systemically to precursors utilized for glycerolipid biosynthesis in the retina.

Animals↗

Effect of light on the metabolism of lipids in the rat retina.

The effect of light on the in vitro incorporation of a variety of radioactive precursors into glycerolipids was tested in isolated retinas of albino rats. There was an increase in the incorporation of [2-3H]myo-inositol, 32Pi, [2-3H]glycerol, and [methyl-3H]choline into retinal phospholipids in light compared to that in darkness. [2-3H]myo-Inositol was incorporated primarily into phosphatidylinositol. 32Pi was incorporated primarily into the phosphoinositides, although there were significant increases in the specific activities of all retinal phospholipids in light compared to those in darkness. Likewise, [2-3H]glycerol incorporation into all retinal phospholipids and diglycerides was greater in light than in the dark. There was no effect of light on the incorporation of [2-3H]ethanolamine into phosphatidylethanolamine or of [3-3H]serine into phosphatidylserine, although these phospholipids were labeled to a greater extent in light with [2-3H]glycerol. There was no effect of light on the incorporation of [3H]palmitic acid into diglycerides and phospholipids, with the exception of phosphatidylinositol. Light also had no effect on the uptake of [2-3H]glycerol, [2-3H]inositol, or [methyl-3H]choline into the retina. We conclude from these studies that light stimulates the phosphoinositide effect in the rat retina. Although some of the results are consistent with a stimulation of de novo synthesis of all lipid classes, our studies with [3H]palmitate, [2-3H]ethanolamine, and [3-3H]serine do not support this conclusion.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effect of lipid peroxidation on rhodopsin regeneration.

We have developed an in vitro system in which lipid peroxidation can be produced in a predictable fashion and have studied the effects of peroxidation on regenerability of rhodopsin in rod outer segments (ROS). ROS isolated by sucrose flotation from dark-adapted retinas of Rana pipiens were suspended in various concentrations of FeSO4 and 1 or 2 mM ascorbic acid for 10 minutes. An increase in lipid hydroperoxides (measured as conjugated dienes) and a decrease in 22:6 omega 3 were determined for each Fe+2 concentration and used as an index of peroxidation. Following incubation with and without FeSO4, ROS were pelleted and rhodopsin was regenerated with 15 micron 11-cis-retinal. The regenerability of rhodopsin in ROS in which significant lipid peroxidation had taken place was reduced by 40-50% compared to controls. DTPA (diethylenetriamine pentaacetic acid), EDTA, and EGTA protected against lipid peroxidation by Fe+2 and allowed almost complete regeneration of rhodopsin. Incubation with DTPA also prevented the destruction of vitamin E in ROS. Other compounds containing primary amine groups did not protect against peroxidation or loss of regenerability. These experiments indicate that lipid peroxidation is associated with a loss of regenerability of the visual pigment rhodopsin.

Animals↗

Biocompatibility analysis of the Sundt-Kees booster clip and the Drake aneurysm clip. Technical note.

A combination of the Sundt-Kees reinforcing aneurysm clip applied to a Drake aneurysm clip in a piggyback fashion was studied for possible defects due to corrosion and or tissue toxicity. These two clips, which are made of similar metal (301 stainless steel), showed little or no defects when immersed in 5% saline or when implanted in rats for 6 months. This study demonstrates that clips made of similar metals can be used piggyback in patients without ill effects provided that they are carefully handled to avoid any abrasion or misbends which could conceivably lead to corrosion.

Animals↗

Manpower needs and supply in academic pathology.

A survey of manpower supply and need in academic pathology departments was conducted by the Joint Task Force on Pathology Manpower of the College of American Pathologists, the American Society of Clinical Pathologists, and the Association of Pathology Chairmen by means of questionnaires distributed to chairmen and faculty of all departments of pathology and laboratory medicine in the United States. Of the 136 departments, responses were obtained from 131 (96%) chairmen and from 1,561 (45%) faculty members. The current total need for faculty members (funded vacant positions) is 272, and the projected aggregate need over the next five years is 693 members for new and replacement positions. The greatest needs are for research MD pathologists and surgical pathologists, followed by autopsy and chemical pathologists. Based on the estimate of current pathology trainees planning academic careers, there is a strong indication that the projected supply of MD pathologists is significantly less than the anticipated demand.

Academic Medical Centers↗

Phosphatidylinositol 4,5-bisphosphate: light-mediated breakdown in the vertebrate retina.

Isolated frog (Rana Pipiens) retinas were labeled in the dark with either [32P] PO4-orthophosphate or myo-[2-3H]inositol for 2.5-4 hrs. After washing the retinas with cold buffer, they were exposed to brief flashes of light (5 secs or 15 secs) and their rod outer segments isolated. Upon separation of labeled phospholipids, a specific decrease in label in phosphatidylinositol 4,5-bisphosphate was observed, whereas there was no significant effect on the labeling of phosphatidylinositol 4-phosphate, phosphatidylinositol, or phosphatidic acid. These results are indicative of a light-activated phosphatidylinositol 4,5-bisphosphate-specific phospholipase C in frog rod outer segments.

Animals↗

Ion-transporting ATPases and matrix mineralization in cultured osteoblastlike cells.

Cultures of osteoblastlike cells obtained from the endosteal surfaces of rabbit long bones formed and mineralized an extracellular matrix when they were supplied daily with medium containing fresh ascorbate. No matrix formed without this supplementation. The matrix mineralized whether or not beta-glycerophosphate, a substrate of alkaline phosphatase, was added to the medium. The ion-transporting ATPase activities of untreated, ascorbate-treated, and ascorbate plus beta-glycerophosphate-treated cells were measured. Ascorbate-treated and ascorbate plus beta-glycerophosphate-treated cells had similar enzyme activities. The activities of the Ca2+-ATPase; Ca2+,Mg2+-ATPase; and alkaline phosphatase in treated cells were elevated over the activities in untreated cells. Na+,K+-ATPase activity was lower in treated than in untreated cells. HCO3--ATPase activity was not changed by treatment. Alkaline phosphatase activity was 20 times higher in freshly isolated osteoblastlike cells than in cells grown to confluence in primary culture. In addition, subculturing further reduced the activity of this osteoblast-marker enzyme. The activities of the ion-transporting ATPases and alkaline phosphatase in second passage cells were similar to the activities of these enzymes in fresh, noncalcifying tissues. Nevertheless, second passage cells retain the ability to mineralize an extracellular matrix, and their ion-transporting ATPase and alkaline phosphatase activities are altered when the cells mineralize a matrix.

Adenosine Triphosphatases↗

Effects of cortisol and fluoride on ion-transporting ATPase activities in cultured osteoblastlike cells.

Na+,K+-ATPase, HCO3(-)-ATPase, Ca2+,Mg2+,-ATPase, Ca2+-ATPase, and alkaline phosphatase activities were measured in cultures of osteoblastlike cells treated with fluoride and cortisol separately and in combinations. Low concentrations of cortisol increased HCO3- -ATPase (10(-11) to 10(-18) M cortisol) and alkaline phosphatase (10(-11) to 10(-9) M cortisol) activities, but higher cortisol concentrations reduced these activities. Na+,K+-ATPase, Ca2+,Mg2+-ATPase, and Ca2+-ATPase activities tended only to be reduced by cortisol. Fluoride (10(-6) and 5 X 10(-6) M) increased HCO3(-)-ATPase and alkaline phosphatase activities, but these activities were similar to controls in the presence of 10(-5) M fluoride. Ca2+,Mg2+-ATPase activity was decreased and Na+,K+-ATPase activity was increased as the concentration of fluoride increased (10(-6) to 10(-5) M). Preliminary experiments with fluoride indicated that lower concentrations (10(-7) M) were without effect. Cortisol concentrations of 10(-9) and 10(-8) M were chosen for studies with combinations of cortisol and fluoride because the effects of these concentrations on alkaline phosphatase activity were opposite, i.e. 10(-9) M increased whereas 10(-8) M decreased activity. Fluoride concentrations of 10(-6), 5 X 10(-6), and 10(-5) M were chosen because a peak of alkaline phosphatase activity occurred at 5 X 10(-6) M fluoride. Higher (10(-4) M) and lower (10(-7) M) fluoride concentrations were without effect. The effects of combinations of cortisol and fluoride depend on the enzyme activity measured. Fluoride (10(-6) M) combined with cortisol (10(-9) M) produced a peak of Na+,K+-ATPase activity. The increased activity obtained with all concentrations of fluoride alone was preserved when fluoride was combined with 10(-8) M cortisol, although the activity tended to be reduced at 5 X 10(-6) and 10(-5) M fluoride. HCO3(-)-ATPase activity was increased by fluoride combined with 10(-8) M cortisol and decreased by fluoride combined with 10(-9) M cortisol compared to the activities obtained with fluoride alone. The decrease in Ca2+,Mg2+-ATPase activity caused by fluoride alone was prevented by 10(-9) and enhanced by 10(-8) M cortisol, although all treatments produced the same activity at 10(-5) M fluoride. Ca2+-ATPase activity tended to be increased by combinations of fluoride and cortisol, but significantly so only at 10(-5) M fluoride in combinations with 10(-8) and 10(-9) M cortisol.(ABSTRACT TRUNCATED AT 400 WORDS)

Adenosine Triphosphatases↗

Inositol incorporation into phosphoinositides in retinal horizontal cells of Xenopus laevis: enhancement by acetylcholine, inhibition by glycine.

The absorption of light by photoreceptor cells leads to an increased incorporation of [2-3H]inositol into phosphoinositides of horizontal cells in the retina of Xenopus laevis in vitro. We have identified several retinal neurotransmitters that are involved in regulating this response. Incubation with glycine, the neurotransmitter of an interplexiform cell that has direct synaptic input onto horizontal cells, abolishes the light effect. This inhibition is reversed by preincubation with strychnine. Acetylcholine added to the culture medium enhances the incorporation of [2-3H]inositol into phosphoinositides in horizontal cells when retinas are incubated in the dark. This effect is inhibited by preincubation with atropine. However, atropine alone does not inhibit the light-enhanced incorporation of [2-3H]inositol into phosphoinositides in the retina. gamma-Aminobutyric acid, the neurotransmitter of retinal horizontal cells in X. laevis, as well as dopamine and norepinephrine, have no effect on the incorporation of [2-3H]inositol into phosphoinositides. These studies demonstrate that the light-enhanced incorporation of [2-3H]inositol into phosphoinositides of retinal horizontal cells is regulated by specific neurotransmitters, and that there are probably several synaptic inputs into horizontal cells which control this process.

Acetylcholine↗

Ethanolamine accumulation by photoreceptor cells of the rabbit retina.

The rabbit retina accumulates ethanolamine by an overall process that has a high affinity for ethanolamine. This process is different from the choline uptake, since ethanolamine accumulation was unaffected by high choline concentrations. Autoradiography identified the major site of high-affinity uptake as the perinuclear region of the photoreceptor cells. Ethanolamine accumulated by the high-affinity uptake was not used for neurotransmission by photoreceptor cells but was used to synthesize phosphatidylethanolamine. However, only a small percentage of the accumulated ethanolamine was converted into phospholipid. The rate of phosphorylation may contribute to control of phospholipid synthesis, since choline kinase activity is much greater than ethanolamine kinase activity in the rabbit retina.

Animals↗

Direct effects of diphenylhydantoin (phenytoin) on the ion-transporting ATPases of cultured osteoblast-like cells.

Diphenylhydantoin (phenytoin) added to cultures of osteoblast-like cells at a concentration of 10 microM, at the high end of the therapeutic range in plasma of unbound phenytoin (4-8 microM), caused reductions in Na+, K+-ATPase and alkaline phosphatase activities and increases in Ca2+-ATPase and HCO3--ATPase activities in homogenates of whole cells and in subcellular fractions of cultured osteoblast-like cells. These data suggest that, if these changes occur in vivo, the osteomalacia associated with treatment with diphenylhydantoin may be mediated in part by direct effects on the ability of bone cells to transport ions.

Adenosine Triphosphatases↗