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Biomedical subjects

K R Wagner

Publications and source records attributed to K R Wagner.

At least 55 records · Page 3Linked to original sources
PubMed1987

Reconstitution of transport of vesicular stomatitis virus G protein from the endoplasmic reticulum to the Golgi complex using a cell-free system.

Transport of the vesicular stomatitis virus-encoded glycoprotein (G protein) between the endoplasmic reticulum (ER) and the cis Golgi compartment has been reconstituted in a cell-free system. Transfer is measured by the processing of the high mannose (man GlcNAc2) ER form of G protein to the man5GlcNAc5 form by the cis Golgi enzyme alpha-mannosidase I. G protein is rapidly and efficiently transported to the Golgi complex by a process resembling that observed in vivo. G protein is trimmed from the high mannose form to the man5GlcNAc2 form without the appearance of the intermediate man GlcNAc2 oligosaccharide species, as is observed in vivo. G protein is found in a sealed membrane-bound compartment before and after incubation. Processing in vitro is sensitive to detergent, and the Golgi alpha-mannosidase I inhibitor 1-deoxymannorjirimycin. Transport between the ER and Golgi complex in vitro requires the addition of a high speed supernatant (cytosol) of cell homogenates, and requires energy in the form of ATP. Efficient reconstitution of export of protein from the ER requires the preparation of homogenates from mitotic cell populations in which the nuclear envelope, ER, and Golgi compartments have been physiologically disassembled before cell homogenization. These results suggest that the high efficiency of transport observed here may require reassembly of functional organelles in vitro.

Animals↗
PubMed1986

Brain metabolic correlates of hypoxic-ischemic cerebral necrosis in mid-gestational sheep fetuses: significance of hypotension.

Mid-gestational sheep fetuses exposed to marked hypoxia for 2 h remain brain intact if MABP is maintained above 30 mm Hg. On the other hand, similarly hypoxic fetuses, if they experience reductions in MABP below 30 mm Hg, develop foci of necrosis that predominantly affect hemispheric white matter and neostriatum. Cortex damage is more restricted and is usually associated with more massive underlying white matter damage. The present study examines the brain metabolic basis for the important role of hypotension in brain injury development in marked hypoxia. Sheep fetuses rendered hypoxic by respiring their ewes with 11% oxygen (fetal PaO2 = 8-12 mm Hg) in which MABP was maintained above 30 mm Hg showed increases in brain lactic acid concentrations to 7-13 mumol/g but unaltered energy charge. In contrast, fetuses that sustained MABP reductions below 30 mm Hg showed increases in lactic acid concentrations in vulnerable structures to 16-24 mumol/g accompanied by marked decreases in energy charge. The vulnerable structures also showed reductions in fructose concentrations but a variable behavior of other brain metabolites including phosphocreatine, glycogen, and glucose. Thus, the present findings suggest a relation between hypotension during marked hypoxia, low energy charge, lactic acid accumulation in brain at high concentrations, and fetal brain injury. The ewes of hypoxic hypotensive fetuses received pentobarbital at lower doses than did those of fetuses that maintained blood pressure. This suggests that pentobarbital plays an important role in protecting the fetal brain from asphyxia by extending the hypoxic fetus's ability to maintain blood pressure in addition to reducing its brain metabolism.

Animals↗
PubMed1986

Hyperglycemia preserves brain mitochondrial respiration during anoxia.

We examined brain mitochondrial function in normo- (5 mM) and hyperglycemic (50 mM) cats after 8 min of anoxia. In anoxic normoglycemic cats, mitochondrial state 3 respiration with NAD-linked substrates glutamate or pyruvate (both plus malate) was inhibited 30-50%. The uncoupler carbonylcyanide p-trifluoromethoxyphenylhydrazone (FCCP) maximally stimulated respiration, indicating that inhibition of phosphorylation, not impairment of electron transport, substrate transport, or oxidation was present. State 3 respiration with succinate (plus rotenone) was unaffected. Mitochondrial respiratory control ratios trended toward reductions whereas ADP/O ratios remained unchanged. In contrast, brain mitochondria from anoxic hyperglycemic cats showed no such inhibition of state 3 respiration and no differences in function from normo- and hyperglycemic control animals except for trends toward loose coupling. Significantly higher brain tissue glucose concentrations were present in hyperglycemic controls as the only metabolite difference compared to normoglycemic controls. At the end of anoxia, hyperglycemic cats exhibited significantly higher cortical lactate and glucose levels but similarly reduced high-energy phosphate concentrations compared to normoglycemic cats. These results demonstrate that increased availability of glucose to gray matter as a consequence of hyperglycemia maintains normal mitochondrial state 3 respiration during exposure to anoxia. Previous survival studies have shown that lower serum glucose concentrations during anoxia are relatively brain protective. This result indicates that the presently described alterations in mitochondrial respiration must be fully reversible.

Animals↗
PubMed1985

Topography of brain metabolites: rhesus monkey, goat, and cat.

This report describes the detailed topographic distribution of metabolites (ATP, phosphocreatine, lactate, glycogen, and glucose) in the brains of three large animal species: rhesus monkey, goat, and cat. The findings demonstrated that adequate fixation of tissue metabolites can be achieved for most brain structures in those animals using in situ freezing. This method, which enables studies of metabolite concentrations in hemispheral and deep brain structures in large animals, can be used to examine correlations between changes in brain metabolism and brain injury in a variety of models of neurologic disease.

Adenosine Triphosphate↗
PubMed1985

Acute changes in regional cerebral metabolite values following experimental blunt head trauma.

A Remington humane stunner was used to deliver a blow to the left side of the surgically-exposed skull in ketamine-anesthetized cats. At 15 minutes after the trauma, brain tissue was frozen in situ. In animals without visible tissue hemorrhage (Grade 0) and in those with unilateral cerebral contusions involving the cerebral cortex and white matter (Grade 2), regional cerebral metabolite concentrations were measured by enzymatic-fluorometric techniques and edema was tested with an organic gradient. No substantial changes in cerebral metabolite concentrations were observed in head-injured animals without cerebral contusions. In animals with unilateral contusions, the white matter neighboring the tissue hemorrhage had an increase in lactic acid and a decrease in phosphocreatine as compared to values from corresponding areas on the contralateral side, and in control and Grade 0 animals. The cerebral cortex adjacent to tissue hemorrhage had a variable response that ranged from metabolite concentrations within normal ranges to marked decreases in high-energy phosphates and increases in lactic acid. Metabolites of the cortex and white matter contralateral as well as distant to contusion were not statistically different from values of control animals. Changes in several metabolites correlated well with the magnitude of edema. It is concluded that focal metabolic alterations can occur shortly after severe blunt head injury, and that these events may contribute to acute traumatic cerebral edema.

Adenosine Diphosphate↗
PubMed1984

A new procedure to analyze free fatty acids. Application to 20-mg brain tissue samples.

Fatty acids were analyzed by a new method which involved their isolation from hexane extracts of serum or brain tissue in aqueous potassium hydroxide (10 microliter) and methylation directly in this solution with methyl iodide. The resulting fatty acid methyl esters were partitioned into ethylene chloride (25 microliter) and were quantitated by gas-liquid chromatography. The procedure was documented by comparison with conventional methylation reactions on serum fatty acids. This method, which avoids thin-layer chromatography and which measures individual free fatty acid concentrations in 20-mg brain tissue samples, should be of particular value for examining regional free fatty acids in brain following ischemia and trauma.

Animals↗
PubMed1983

Staphylococcus aureus meningitis: 26 years' experience at Vancouver General Hospital.

The records of all patients with Staphylococcus aureus meningitis admitted to Vancouver General Hospital between 1956 and 1981 were reviewed. All the patients had clinical and laboratory features of meningitis, and in all cases S. aureus was isolated from the cerebrospinal fluid. S. aureus was responsible for 21 (3%) of the 710 cases of acute bacterial meningitis. Therapy with cloxacillin or methicillin, or both, with or without other agents, was successful in 14 of the 21 patients. Three of the 14 patients without ventricular shunts died, 2 with fulminating septicemia and 1 with a postoperative brain abscess treated with cloxacillin. Following shunt removal and antibiotic therapy all seven patients with ventricular shunts survived the infection. Shunt removal may therefore be essential in appropriate cases.

Cerebrospinal Fluid Shunts↗
PubMed1978

The pentose phosphate pathway in regenerating skeletal muscle.

1. The activities of the oxidative enzymes (glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase) and of the non-oxidative enzymes (transaldolase, tranketolase, ribose 5 phosphate isomerase and ribulose 5-phosphate 3-epimerase) of the pentose phosphate pathway were measured at various times during the first 24h of skeletal-muscle regeneration after administration of Marcaine, a mytoxic local anesthetic. 2. The activities of the oxidative enzymes increased after Marcaine injection and rose to 9 times control activities by 24h. 3. The activities of all non-oxidative enzymes were increased after Marcaine administration, but to a much smaller extent than the oxidative enzymes (1.1-1.7-fold). 4. Histochemical analysis localized glucose 6-phosphate dehydrogenase activity within muscle fibres of control and Marcaine-treated muscles. 5. Cycloheximide or actinomycin D prevented the increase in oxidative enzyme activities, suggesting a requirement for synthesis of protein and RNA.

Animals↗
PubMed1977

Developmental patterns of glycolytic enzymes in regenerating skeletal muscle after autogenous free grafting.

Extensor digitorum longus muscles of rats were removed and injected with a solution of Marcaine plus hyaluronidase. After incubation in Marcaine solution for 10 min, the muscles were grafted into their original beds. The grafts and the contralateral control muscles were removed from the rats at 0, 1-5, 7, 11, 36, and 69 days postoperatively. The muscles were then frozen in dry ice and isopentane and subsequently homogenized and centrifuged. The supernatant was analyzed for a number of enzymes, the regenerative patterns of which can be classified into 3 groups: (1) early increase in activity: hexokinase, glucose-6-phosphate dehydrogenase; (2) early decrease in activity with failure to recover to control levels: phosphorylase, phosphofructokinase, alpha-glycerophosphate dehydrogenase; and (3) early decrease followed by return to control levels: lactate dehydrogenase, pyruvate kinase, creatine phosphokinase, adenylate kinase. These patterns are not identical to those reported for embryogenesis of muscle. The data are discussed with regard to correlative histological studies of muscle regeneration.

Adenylate Kinase↗
PubMedDate not supplied

Reinnervation of rat extensor digitorum longus muscles after free grafting.

Free grafts of the extensor digitorum longus (EDL) muscle of the rat are invaded by regenerating nerve fibers during the second postoperative week. Neuromuscular junctions are established during the third week. Levels of choline acetyltransferase (CAT) activity parallel the invasion of the graft by nerve fibers, but this activity never returns to control levels. We conclude that incomplete reinnervation of free muscle grafts may be a factor in the failure of such grafts to reattain the mass of control muscles.

Animals↗
PubMedDate not supplied

Stroke models: strengths and pitfalls.

Recent editorials and reviews express disillusionment and sharp criticism with the contribution of animal experimental studies to stroke prevention and treatment. The basis for these comments appears to be a frustration with the absence of pharmacologic agents to effectively treat stroke patients despite considerable research efforts. In response to these nihilistic views, Zivin and Grotta (Stroke, 21 (1990) 981-983) have written a poignant rebuttle and Goldstein (Stroke, 21 (1990) 373-374) has outlined the considerable progress which has been achieved in experimental cerebral stroke research. It is our goal in the present discussion to highlight the strengths and to examine the potential pitfalls of stroke models, some of which may have contributed to the equivocal results obtained in testing pharmacologic agents. In addition, based on our own experience with a model of middle cerebral artery (MCA) occlusion in cats, we will describe the benefits of large versus small animal models for experimental stroke studies.

Animals↗