Journal of Histochemistry & Cytochemistry

HOLTZMAN, ERIC

doi: 10.1177/13.5.318pmid: 14344249

Evidence for heterogeneity in solubility properties of the histones of fixed Necturus liver was obtained by extracting freeze-methanol-substituted tissue with 2-10% perchloric acid, ethanol-HCl, or 0.02 N HCl, postfixing in formaldehyde then staining with the Alfert-Geschwind method for basic proteins and measuring the dye bound to nuclei by microspectrophotometry.Data was obtained which can be interpreted as indicating that each of the acids extracts a discrete portion of the histone. Information from experiments based on sequential extractions with two different reagents suggests that the portions extracted may represent different subfractions, perhaps corresponding to histone subtypes differing in relative content of arginine and lysine and in other properties.

JENNINGS, BARBARA M.

doi: 10.1177/13.5.328pmid: 14344250

Aldehyde-fuchsin staining was applied to fresh frozen sections to frozen sections pre-fixed with sucrose-formol, and to freeze-dried sections of human and animal pancreas and pituitary, with positive reactions in β-cells of both tissues. Successful adaptation of aldehyde-fuchsin staining to this material was achieved largely by choice of fixative, use of a fresh, nonchilled solution of aldehyde-fuchsin, and by acidified permanganate oxidation.Certain standard fixatives (formol, Zenker-formol, and Bouin) proved satisfactory, as well as a more recently introduced one, glutaraldehyde. A fixation-staining technique requiring less than 10 minutes was evolved using stock (25%) glutaraldehyde.With some of these fixatives, inserting an aldehyde blockade step when necessary, the Periodic and Schiff reaction could be employed as a control stain for aldehyde-fuchsin, with comparable results.The methods adopted are useful in identifying β-cells in surgical specimens and in material used for enzyme reactions and fluorescent antibody techniques.

GAHAN, P. B.

doi: 10.1177/13.5.334pmid: 14344251

Using the enzyme acid phosphatase as a marker for lysosomes, a histochemical study has been made of the effects of the operative procedure for partial hepatectomy upon the lysosomes of rat liver. The procedure was considered in three stages: i) ether anaesthesia; ii) laparotomy; iii) laparotomy and handling of the left and ventral lobes. With increasing severity of procedure shorter times of incubation in test medium were required to demonstrate lysosomes in liver. This was considered to indicate an increased permeability of lysosomal membrane to the substrate, β-glycerophosphate. Response was not immediate, but increased to a maximum with time, finally returning to normal. The reversible change in lysosomes was considered to reflect a physiological response by these particles to cell damage.

COUTINHO, HÉLIO B.; PADILHA, MANOEL C. S.; GOMES, JURANDY M.; ALVES, J. J. ALMEIDA

doi: 10.1177/13.5.339pmid: 14344252

A comparative study was made of the liver, kidney, and lung esterases of 20 different mammals, using the starch gel zymogram method. Differences in the organs of each species and the particular zymogram pattern of each animal are pointed out. The results obtained cannot be correlated with the phylogenetic position of the species studied.

AVERS, C. J.; LIN, F. H.; PFEFFER, C. R.

doi: 10.1177/13.5.344pmid: 14347365

Histochemical localizations of cytochrome oxidase, succinic dehydrogenase, DPNH-tetrazolium reductase, and TPNH-tetrazolium reductase activities revealed at least two kinds of mitochondria in the intracellular population. The total chondriome in stationary phase cells contains about 45 mitochondria, all with cytochrome oxidase activity. But, only about 30 mitochondria per cell were active for dehydrogenase or reductases. The differences in mitochondrial enzyme activities persisted throughout the growth cycle, showing different numbers of active mitochondria and different rates of their increase and decrease for all four enzyme systems. Manometric data verified the differences between cytochrome oxidase and succinic dehydrogenase for the earlier phases of the growth cycle. In histochemical counts, zero values for all four enzymes occurred in late acceleration phase, but persisted into log phase only for the tetrazolium reductases. Both cytochrome oxidase- and succinic dehydrogenase-active mitochondria began to increase in numbers at the inception of log phase, but at very different rates. The demonstration of more than one kind of mitochondrion in the common nucleocytoplasmic system of a single cell was considered to be evidence of some measure of autonomous control of the mitochondrial phenotype.

BOSS, J. H.

doi: 10.1177/13.5.350pmid: 14344253

The microscopic immunofluorescence of the glomeruli differs in acute and chronic nephrotoxic serum nephritis in the distribution pattern of the injected nephrotoxic globulin and the host's gamma globulin. In acute nephritis specific glomerular fluorescence, denoting fixation of nephrotoxic and recipient's gamma globulin, is sharply limited to regular, thin and delicate lines corresponding to the capillary basement membranes. On the other hand, specific glomerular fluorescence in chronic nephritis appears as irregular, broad and tortuous loops consisting of coarse, partially coalescing specks of differing brightness. This difference is possibly related to the profound alterations in chronic nephritis resulting in reconstruction of the glomerular architecture.

HAYASHI, MASANDO

doi: 10.1177/13.5.355pmid: 14344254

The N-acetyl-β-glucosaminide of naphthol AS-BI (7-bromo-3-hydroxy-2-naphth-o-anisidide) was obtained by reacting the anisidide with acetochloroglucosamine in aqueous alkaline acetone. After removal of O-acetyl groups with methanolic ammonia and recrystallization from aqueous methanol, the compound may be used as a substrate for N-acetyl-β-glucosaminidase.Formol-calcium fixed frozen sections were incubated with 0.5 mM substrate in the presence of hexazonium pararosanilin or fast garnet GBC at pH 5.2. Staining appeared in the cytoplasm mostly as discrete granules at the presumed site of enzyme activity and was inhibited in the presence of N-acetylglucosaminolactone. In the rat kidney a number of spherical granules which stained for the enzyme were observed in the proximal portion of the proximal convoluted tubules. In the liver staining was recognized as small granules localized at the pericanalicular cytoplasm of parenchymal hepatic cells. The general cytological localization in these tissues was quite similar to that of other lysosomal enzymes. The limitations of the technique were briefly discussed.

QUINTARELLI, GIULIANO; DELLOVO, MARIE C.

doi: 10.1177/13.5.361pmid: 14347366

The authors have studied the effects of testosterone propionate on the lacrimal gland of the mouse. Repeated injections of the hormone have determined some structural and functional modifications of the gland. The former were characterized by cytoplasmic vacuoles; the latter by an increase of glycoprotein material.The androgen used seemed to stimulate the synthesis of high molecular weight carbohydrates in a tissue that per se is mainly protein secreting.