Journal of Histochemistry & Cytochemistry

SELYE, HANS; GABBIANI, GIULIO; SERAFIMOV, NIKOLA

doi: 10.1177/12.8.563pmid: 14209993

Experiments were performed on rats, using special histochemical stains for the demonstration of metachromatic materials, lead, calcium, phosphate and carbonate, to study calcergy, the induction of soft-tissue calcification by topical treatment with certain compounds such as lead acetate.In simple calcergy, when lead acetate is directly injected into the subcutis, it is seen to impregnate the collagen fibers in the injection site without noteworthy participation of the mast cells. The lead-treated area secondarily attracts calcium, phosphate and possibly carbonate.In mastocalcergy, the lead acetate is injected intravenously and local calcification is produced by the simultaneous subcutaneous injection of a mast-cell discharger such as polymyxin. Here, the mast cells in the treated area first show degranulation without any mineralization, but this is soon followed by lead uptake on the mast-cell granules with secondary attraction of calcium, phosphate and possibly carbonate. Finally, the discharged and calcified mast-cell granules disintegrate into a dustlike, fine precipitate which is transferred to the circumjacent collagen fibers where it initiates an intense process of mineralization, eventually leading to the complete petrification of the polymyxin-injected area.Apparently, in addition to its already well established functions of histamine, heparin and serotonin production, the mast cell also plays a role in mineralization.

ERÄNKÖ, OLAVI; HÄRKÖNEN, MATTI; KOKKO, AULIKKI; RÄISÄNEN, LIISA

doi: 10.1177/12.8.570pmid: 14209994

CORRODI, HANS; HILLARP, NILS-ÅKE; JONSSON, GÖSTA

doi: 10.1177/12.8.582pmid: 14209995

It has been shown in previous papers that catecholamines and 5-hydroxytryptamine can under certain conditions be converted to highly fluorescent 6,7 - dihydroxy - 3,4 - dihydroisoquinolines and 6-hydroxy-3,4-dihydro-β-carboline respectively, and that this can be used as a highly sensitive and specific method for the histochemical demonstration of the monoamines at the cellular level. In the present paper it is shown that the fluorescent compounds are very readily reduced by sodium borohydride to the corresponding, non-fluorescent 1,2,3,4-tetrahydro-compounds—even if they are present in a non-extractable state in dried serum albumin spots or in tissue sections—and that the fluorescence can be regenerate by renewed formaldehyde treatment. The non-specific fluorescence (e. g. autofluorescence) in tissue sections was never observed to undergo any changes on borohydride treatment. On the basis of these findings a very simple histochemical test has been worked out to check directly in the tissue section whether or not an observed fluorescence is due to the presence of the reacting monoamines.

AMBADKAR, P. M.; GEORGE, J. C.

doi: 10.1177/12.8.587pmid: 14209996

The localization and distribution of β-hydroxybutyrate dehydrogenase, succinic dehydrogenase, malic dehydrogenase, and lactic dehydrogenase in the rat testis have been studied histochemically. Intense enzyme activity in the interstitium as well as tubules was observed. However, malic dehydrogenase was found to be more active in the interstitium than in the tubules. The distribution pattern of enzyme activity appeared in four different phases in the seminiferous tubules corresponding to the gradient in the spermatogenetic wave, thereby indicating a metabolic adaptation at subcellular level.

KENT, SIDNEY P.

doi: 10.1177/12.8.591pmid: 14209997

A fluorescein labelled extract of Ulex europeus seed was found to be satisfactory for demonstrating water soluble H antigen in formalin fixed paraffin embedded tissue sections. Commercially available anti-A typing serum was labelled with fluorescein and used to demonstrate water soluble A antigen in formalin fixed paraffin embedded tissues. The reactivity of A and H antigen after neutral buffered formalin fixation was superior to that observed after Zenker's, Helly's or Bouin's fixation; reactivity in paraffin sections of formalin fixed tissues were equal to that of fresh frozen sections. Autolysis did not prevent the demonstration of blood group antigen A or H in fixed paraffin embedded tissues but was associated with some diffusion of the antigens.The distribution of H antigen in formalin fixed paraffin embedded tissue section for patients with type A, B and O blood also was studied. In secretors blood group substances were widely distributed in the epithelial mucins of the body whereas in nonsecretors the antigens are largely confined to the deep portion of the pyloric mucosa of the stomach and Brunner's glands. The distribution of A and H antigen within a gland, such as Brunner's glands, was not homogeneous. That is, some acini contained A antigen but no H antigen; others contained H antigen but no A antigen: a few contained both A and H antigen. The significance of this mosaic distribution which previously has not been described was discussed.

KELLY, JOHN W.; CLABAUGH, W. A.; HAWKINS, H. K.

doi: 10.1177/12.8.600pmid: 14209998

Improved apparatus for dual-microscope photographic cytophotometry is described. The major component is a new comparison microscope, available commercially, whose binocular system is optically corrected for performance equivalent to that expected of one microscope. While current applications involve only standard bright-field and interference optics for absorption and dry-mass studies, a feature of the assembly is ready interchangeability of optics for fluorescence, polarization, phase, and dark-field microscopy.A method for extraction of color-film dyes into dimethylsulfoxide is reported. The dye solutions are used for spectrophotometric measurement of amount of dye in a given area of film and, indirectly, for estimating the amount of chromophore in the corresponding area of a microscopic object.

VAN WIJHE, M.; BLANCHAER, M. C.; ST. GEORGE-STUBBS, S.

doi: 10.1177/12.8.608pmid: 14209999

A study of the distribution of lactate dehydrogenase isozymes in single fibers from normal human skeletal muscle is presented. The fibers were classified into red, intermediate and white types on histochemical grounds and their lactate dehydrogenase isozyme content assessed by electrophoretic separation in veronal buffered agar. The results generally agreed with previous homogenate studies on animal skeletal muscle, in that the white fibers contained almost exclusively isozymes IV and V, whereas red fibers were rich in isozymes I, II and III, but IV and V also appeared indigenous to these fibers. The intermediate fibers had an isozyme pattern combining the features of red and white fibers. The metabolic implications of these findings are discussed.

O'STEEN, W. KEITH; DILL, RUSSELL E.

doi: 10.1177/12.8.615pmid: 14210000

Outgrowth of rat pineal tissue in diffusion chambers began with a migration of connective tissue cells from the implant, and this was followed by a spreading of the parenchymal cells to the margins of the chambers.Diffusion chamber implants and fresh slices of pineal organ were incubated for monoamine oxidase (MAO) in a medium containing nitro-blue tetrazolium and tryptamine hydrochloride. The MAO reaction was visualized as black formazan granules in parenchymal cells of all cultures throughout the entire 21 day observation period. These cytoplasmic granules appeared to increase in number and in size during the later time periods. A positive enzyme reaction also was present in slices of fresh pineal organ treated in the same medium and in a medium without the substrate, tryptamine hydrochloride. Pineal tissues with prior treatment in a MAO inhibitor gave a negative reaction.Some parenchymal cells in diffusion chamber implants accumulated many large lipid droplets. There were fewer of these cells than MAO positive cells, and they appeared to migrate from the implant at a slower rate.

VILAR, O.; ALVAREZ, B.; DAVIDSON, O.; MANCINI, R. E.

doi: 10.1177/12.8.621pmid: 14210001

Several Armour and NIH pituitary hormone preparations as well as other homologous serum protein fractions were labeled with fluorescent dyes (sulphorhodamine B, C.I. 45100, and fluorescein isothiocyanate). Rats were injected intravenously with the different labeled proteins and the animals killed at various intervals from 3 minutes to 10 days. Sections of the kidneys were studied under ultraviolet light (3,900 Å). It was observed that all the labeled proteins passed the glomerular filter and were accumulated by the cells of the proximal convoluted tubules. Labeled pituitary hormones were, in addition, reabsorbed by the thin limb and collecting tubules and appeared in the lumen of bladder. Filtration and reabsorption of proteins by the kidney can proceed independently of their nature and origin. All the proteins were reabsorbed with the same sequence, but pituitary hormones appeared faster and in larger amounts in the same sites of the nephron. They were also detected in the lumen of the ureter and in the bladder. It is assumed that the molecular weight of the proteins may play an important role, since the hormones' molecules were significantly smaller. It is remarkable that although the different hormones showed a preferential accumulation in the vessels of the corresponding target glands, they behave similarly in the kidney.

MARTAN, JAN; ALLEN, JOHN M.

doi: 10.1177/12.8.628pmid: 14210002

Holocrine secretory cells have been identified in the epithelium of the epididymal canal of the mouse. These cells develop from basal cells. During their differentiation they grow toward the lumen of the epididymal canal and come to form club-shaped structures with an expanded apical portion, a central nucleus and a thin stalk-like connection to the basement membrane. Mature holocrine cells are characterized by their high acid phosphatase and aliesterase activity. They also are highly active for succinic dehydrogenase, nicotinamide adenine dinucleotide diaphorase, and nicotinamide adenine dinucleotide phosphate diaphorase. Nucleoside diphosphatase, thiamine pyrophosphatase, adenosine triphosphatase, and alkaline nucleoside phosphatase are also found in these cells. These cells are also characterized by their reactivity with the Aoyama and periodic acid-Schiff reactions. They react moderately with the molybdate and Luxol Fast Blue MBS reactions for choline containing compounds. Mature holocrine cells may disintegrate in situ or may be discharged in toto into the lumen of the epididymal canal. Glycerylphosphorylcholine was identified in extracts prepared from sperm-free epididymides of mice. Glycerylphosphorylcholine reacts with Aoyama and periodic acid-Schiff reactions as do mature holocrine cells. This fact coupled with the identification of choline containing material in holocrine cells suggests that they may be one site for the formation of glycerylphosphorylcholine.