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Biomedical subjects

A B Johnson

Publications and source records attributed to A B Johnson.

101 records · Page 6Linked to original sources
PubMedDate not supplied

On-grid immunogold labeling of glial intermediate filaments in epoxy-embedded tissue.

On-grid immunogold labeling of structures like intermediate filaments has been difficult to achieve. Presumably this is because such structures are thinner than the thin sections themselves and because gold-labeled reagents remain on the surface and do not penetrate epoxy resins. Many pathologic and other tissues, however, are primarily available as epoxy-embedded blocks, and a postembedding gold procedure capable of detecting such thin structures would be useful. This study aimed to investigate the astrocytic intermediate filament antigen glial fibrillary acidic protein (GFAP) in glutaraldehyde-fixed, epoxy-embedded brain biopsy tissue from a child with Alexander's disease. A protocol was developed for performing on-grid immunogold labeling which minimized nonspecific deposition of gold reagent. The method utilized ovalbumin and skim milk in the washes and diluent for the gold reagent and the same solution with added Tween-20 and high sodium chloride in the diluent for antibodies and normal serum. In grids etched with metaperiodate and hydrogen peroxide, the astrocytic intermediate filaments were only occasionally and sparsely labeled. When an etching procedure with sodium ethoxide was employed, however, extensive labeling was obtained on the astrocytic intermediate filaments. In contrast, the larger, pathological Rosenthal fibers characteristic of Alexander's disease were labeled after both etching procedures, but labeling was enhanced after ethoxide etching. Postosmicated material showed much less labeling. The findings demonstrate that postembedding procedures can be used with epoxy-embedded material to immunolabel thin structures like intermediate filaments.

Cytoskeleton↗
PubMedDate not supplied

Detection of reovirus type 3 in the porta hepatis of an infant with extrahepatic biliary atresia: ultrastructural and immunocytochemical study.

This report describes immunocytochemical and ultrastructural methods which led to the identification of Reovirus type 3 (Reo-3) in the porta hepatis of a patient with extrahepatic biliary atresia. The study indicates that Reo-3 antigenic sites are demonstrable by the avidin-biotinylated complex peroxidase method following formalin fixation and paraffin embedding, but are destroyed by freezing and thawing prior to fixation. Deparaffinization of the block and subsequent rembedding in epon-araldite did not alter immunoperoxidase staining. This procedure offered the advantage of higher light microscopic resolution of semithin (1 micron) sections and assisted in the selection of specific areas for ultrastructural studies. Localization of Reo-3 in extrahepatic biliary atresia was confined to a biliary remnant in which there were acutely inflammed, partially necrotic microscopic ducts. Electron microscopic examination of the immunoreactive sites revealed virus-like particles similar in appearance to Reo-3 particles in infected tissue culture cells. The observations presented here support previously reported serologic data which have shown an association between Reo-3 infection and extrahepatic biliary atresia.

Animals↗
PubMedDate not supplied

Exercise augments the absorption of isophane (NPH) insulin.

In order to determine the effect of exercise on the rate of absorption of an isophane (NPH) insulin, 7 normal men were studied on two separate occasions using the euglycaemic clamp technique. On one day subjects undertook 60 min of exercise on a treadmill (5 km h-1, 5 degrees slope) 180 min after injection of human isophane (NPH) insulin (0.25 U kg-1), while on the control day they remained at rest throughout the 420 min study. Serum insulin concentrations rose in parallel after injection achieving similar values at 180 min on exercise and control days (13.7 +/- 1.2 (+/- SE) vs 12.0 +/- 1.5 mU l-1; NS), respectively. After the onset of exercise, insulin concentrations rose markedly to a peak of 29.8 +/- 2.2 mU l-1 after 50 min of exercise (p less than 0.001), compared with a slight fall on the control day to 10.4 +/- 1.7 mU l-1 (NS). At the end of the exercise period, serum insulin concentrations returned rapidly to control day values. Glucose infusion requirements rose significantly during the exercise period from 2.8 +/- 0.5 mg kg-1 min-1 to a peak at 235 min of 11.1 +/- 1.2 mg kg-1 min-1 (p less than 0.001), compared with an increase on the control day from 2.0 +/- 0.6 to 2.5 +/- 0.6 mg kg-1 min-1 (NS) over the same period. These results demonstrate an enhanced rate of absorption of an isophane (NPH) insulin during exercise in normal subjects.

Adult↗
PubMedDate not supplied

The effect of intravenous metformin on glucose metabolism during hyperglycaemia in type 2 diabetes.

A stepped intravenous metformin infusion was used in conjunction with the hyperglycaemic clamp technique to study the dose-response relationship of plasma metformin concentration with hepatic glucose production and peripheral glucose disposal in nine patients with Type 2 diabetes. The study was of double-blind crossover design, using NaCl infusion as control. Plasma metformin concentrations spanning the therapeutic range (1.64 +/- 0.13 mg l-1 and 6.57 +/- 0.61 mg l-1) were achieved. No differences in peripheral glucose disposal were demonstrated when compared with NaCl infusion (3.4 +/- 0.1 vs 3.6 +/- 0.2 (+/- SE) mg kg-1 min-1 and 3.4 +/- 0.2 vs 3.3 +/- 0.2 mg kg-1 min-1, respectively). There was also no difference in basal hepatic glucose production during metformin and NaCl infusion (2.7 +/- 0.3 vs 2.8 +/- 0.2 mg kg-1 min-1). No acute effect of metformin on hepatic glucose production or peripheral glucose disposal was observed, implying that a chronic persistent effect is more important in these respects than immediate effects consequent upon changes in plasma drug level.

Blood Glucose↗
PubMedDate not supplied

A correlative study of the adrenal cortex in adreno-leukodystrophy--evidence for a fatal intoxication with very long chain saturated fatty acids.

Thirty adrenal glands from patients with adreno-leukodystrophy (ALD) have been studied by light microscopy, three by enzyme histochemistry, three by electron microscopy and two by tissue culture. Cytoplasmic ballooning and striations result from proliferation of smooth endoplasmic reticulum and accumulations of lamellar-lipid profiles and clear clefts (crystalloids). Striated adrenocortical cells, the only pathognomonic adrenal lesion in ALD, display cytoplasmic lamellae, decreased amounts of rough endoplasmic reticulum and depression of several enzymes (alpha-glycerophosphate dehydrogenase, 3 beta-hydroxysteroid dehydrogenase and TPNH diaphorase). The striated cells also demonstrate decreased ability to adapt to changes in microenvironment, both in vivo and in vitro. A blunted response by striated cells to focal peripheral cytolysis leads to cytoplasmic erosion, atrophy and macrovacuoles. ACTH has a pivotal role in the evolution of these lesions. We propose that the pathognomonic lamellae of ALD basically represent bilayers or bimolecular leaflets of very long chain saturated fatty acids, while lamellar-lipid profiles and clefts contain cholesterol esterified to these abnormal fatty acids. The similarity of lamellar-lipid profiles of ALD to cytoplasmic lesions induced by long chain saturated fatty acids suggests that the very long chain saturated fatty acids isolated in ALD are cytotoxic and are responsible for adrenocortical cell dysfunction in this disease.

Adrenal Cortex↗