bluuebunny/crossref_metadata_2025_split huggingface dataset:
HISTOCHEMICAL AND ULTRASTRUCTURAL STUDIES ON HELA CELL CULTURES EXPOSED TO SPINDLE INHIBITORS WITH SPECIAL REFERENCE TO THE INTERPHASE CELL
Authors: ROBBINS ELLIOTT , GONATAS NICHOLAS K. 
Published: April 2011

Mammalian cells of the HeLa (S3) strain, when exposed to spindle inhibitors, have been found to undergo several morphological transformations during interphase as well as during mitosis. Some of these have been studied both histochemically and ultrastructurally. The lysosomes, represented by the multivesicular bodies in HeLa cells, form clusters and become circumferentially disposed instead of occupying the polarized juxtanuclear position characteristic of these organelles. In the electron microscope it is seen that they have acquired a dense osmiophilic core that is separated from the bounding unit membrane by an electron lucent halo. The Golgi apparatus fragments under the influence of spindle inhibitors and also takes up a circumferential distribution in a pattern similar to that of the lysosomes. On the ultrastructural level, no significant modifications in this organelle are seen. Also noted in the interphase cell are numerous 60-80 Å fibrils coursing through the cytoplasm as well as a paucity of spindle our microtubules. A striking similarity has been pointed out between the behavior of the lysosomes in the drugtreated interphase cell and the untreated, normal mitotic cell. A possible explanation of some of the changes noted has been given in terms of an interruption of protoplasmic flow resulting from the disappearance of microtubules.
DIAMINE METHODS FOR DIFFERENTIATING MUCOSUBSTANCES HISTOCHEMICALLY
Authors: SPICER S. S. 
Published: April 2011

A mixture of N,N-dimethyl- m-phenylenediamine (HCI)2 with the para isomer at pH 3.5-4.0 stains acid mucosubstances selectively. Various sialomucins as well as sulfomucins differ in their reactivity with this mixed diamine reagent and in the manner in which the pH and ionic strength of the solution affect their staining. Prior periodate oxidation eliminates or markedly decreases the staining of some sulfo- and sialomucins but has no influence on the reactivity of others. Both sulfated mucosaccharides and sialomucins differ also according to whether or not their affinity for azure A, colloidal iron or alcian blue persists following periodate oxidation and exposure to the meta diamine. Azurophilia of nuclei at pH 4.0 is abolished by meta diamine used after Feulgen hydrolysis. Ferric chloride added to a solution of both diamines allows selective demonstration of most acid mucosubstances. This "low iron" diamine method followed by alcian blue stains sulfomucins and many sialomucins black, but other sialomucins blue. Some epithelia contain a mixture of blue and black-stained sialomucins. A variant technique uses a higher concentration of iron and diamines. This colors most sulfomucins black while leaving nonsulfated acid mucosubstances unstained. This high iron diamine method followed by alcian blue stains sulfomucins black and sialomucins blue. Results with the "high iron" diamine-alcian blue sequence indicate that sulfomucin occurs in some sites and sialomucin in others; in many epithelia a mixture of the two mucosubstances coexists either in the same on in different cells. Similar differentiation was obtainable also in many but not all sites with either the combination aldehyde fuchsin-alcian blue procedure on by assessment of differences between the results of alcian blue pH 2.5-PAS and alcian blue pH 1.0-PAS sequences. Applied to the surface mucous layer of certain intestinal and genitourinary epithelia, the methods differentiate areas of sulfomucin content alternating with others of sialomucin content. This is believed to indicate biogenesis of this secretion in the Golgi zone of nongoblet epithelial cells since staining of the surface mucous layer parallels that in the Golgi zone and often not that of neighboring goblet cells.
ENZYME HISTOCHEMISTRY OF BONE INDUCTION BY URINARY BLADDER EPITHELIUM
Authors: KAGAWA SHOHEI 
Published: April 2011

Homogenous transplants of urinary bladder mucosa were made in guinea pigs, and induced bone formation was observed histochemically for alkaline phosphatase, acid phosphatase, esterase, β-glucuronidase, aminopeptidase, and oxidative enzymes, i.e., succinic dehydrogenase, diphosphopyridine nucleotide-dependent dehydrogenase (lactate, malate, glutamate, α-glycerophosphate, β-hydroxybutyrate) and triphosphopyridine nucleotide-dependent dehydrogenase (glucose-6-phosphate and isocitrate). Normal urinary bladder epithelium contained intense alkaline phosphatase and slight acid phosphatase activity throughout. There was weak esterase activity in intermediate layer and weak β-glucuronidase activity in intermediate layer. Succinic dehydrogenase was present throughout the epithelium, and was most active in the basal layer. Lactic and malic dehydrogenase activities were intense. Glutamic, α-glycerophosphate and β-hydroxybutyric dehydrogenase activities were low, but glucose-6-phosphate and isocitric dehydrogenase activities were high. In the initial stage after transplantation, alkaline phosphatase, aminopeptidase and lactic dehydrogenase appeared in the connective tissue surrounding the transplanted mucosa in association with an inflammatory infiltration. Epithelial transplants formed cysts. Lactic, malic and triphosphopyridine nucleotide-dependent dehydrogenases in cystic epithelium were as intense as in normal bladder, though other enzymes decreased. Hyaline formation occurred around the cyst. No appreciable enzyme activity was demonstrated in this hyalinized portion, but when bone appeared marked activity of alkaline and acid phosphatases was seen around it. Histochemical patterns in the induced bone were essentially the same as in normal bone.
A CYTOCHEMICAL STUDY OF THE SOLUBILITIES OF THE HISTONES OF FIXED NECTURUS LIVER
Authors: HOLTZMAN ERIC 
Published: April 2011

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.
ACTIVATION OF GLYCOPROTEIN BIOSYNTHESIS BY TESTESTERONE PROPIONATE ON MOUSE EXORBITAL GLANDS
Authors: QUINTARELLI GIULIANO , DELLOVO MARIE C. 
Published: April 2011

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.
SELECTIVE BINDING OF ZINC BY BASIC PROTEINS OF THE HELA CELL NUCLEOLUS
Authors: STUDZINSKI GEORGE P. 
Published: April 2011

Treatment of ethanol-fixed HeLa cells with solutions containing zinc ions results in marked retention of zinc by the nucleolus. Subsequent exposure to dithizone results in selective staining of the nucleolus with visualization of some of its internal structure. Free amino groups of protein are shown to be chiefly responsible for binding of the zinc, but the imidazole rings of histidine residues may also play a part in zinc retention. High concentrations of protein-bound amino groups can be demonstrated by the ninhydrin-Schiff or the hydroxynaphthaldehyde methods to have a distribution in the HeLa cell nucleolus similar to the sites of marked zinc retention, and the Pauly reaction shows that imidazole groups of histidine are concentrated in the nucleolus in a similar fashion. The zinc-binding protein is acid extractable and it is suggested that this protein may represent a product of the nucleolus with functions in other parts of the cell, where the reactive basic groups are masked by linkages to acidic groups, perhaps of nucleic acids.
THE EFFECT OF OVARIAN HORMONES ON THE SULPHOMUCOPOLYSACCHARIDES OF THE UTERUS
Authors: BO W. J. , SMITH S. , REITER R. , PIZZARELLO D. J. 
Published: April 2011

The distribution of sulphomucopolysaccharides was determined radioautographically and the distribution of acid mucopolysaccharides was determined metachromatically in the uteri of hormone-treated ovariectomized rats. The animals were autopsied 12, 90, and 204 hours after itraperitoneal treatment with sodium sulphate-S35 and a definite difference in the uptake of labelled material was noted in the uteri of the hormone-treated animals autopsied 204 hours after administration of S35. The greatest variability was observed in the endometrial stroma. Radioactive sulphur was scattered lightly throughout the stroma of the ovariectomized and ovariectomized-progesterone treated rats, while in the ovariectomized-estrogen treated animals the silver grains were extensive and localized primarily in the outer part of the stroma. In the estrogen-progesterone treated rats more grains were present than in the castrate, but less than seen following estrogen alone. The silver grains were observed over the extracellular matrix and over the connective tissue cells. Metachromasia was more intense and widespread in the endometrial stroma following estrogen treatment than in the ovariectomized and ovariectomized-progesterone treated animals. The present study indicates that uterine sulphomucopolysaccharides are influenced by the ovarian hormones and that the concentration is highest following estrogen stimulation.
THE EFFECTS OF ALDEHYDE FIXATION ON ACID PHOSPHATASE ACTIVITY IN TISSUE BLOCKS
Authors: JANIGAN DAVID T. 
Published: April 2011

The effect of various aldehydes on phosphatase(s) of rat liver and kidney hydrolyzing the monophosphates of β-glycerol, phenol, p-nitrophenol and naphthol AS-TR at pH 5.0 was determined. Biochemical data were correlated with histochemical staining results. On the basis of enzyme recovery after fixation, the aldehydes could be divided into 3 broad groups: (1) well over 50 per cent after hydroxyadipaldehyde and glyoxal; (2) near 50 per cent after formaldehyde and methacrolein; and (3) well below 50 per cent after crotonaldehyde, glutaraldehyde, and acrolein. Acid phosphatase(s) hydrolyzing all 4 substrates showed a similar differential response to fixation by 4 aldehydes, but individual recovery values with naphthol AS-TR phosphate were distinctly lower. Tissues immersed in the first aldehyde group were soft, showed poor morphologic preservation, and a large percentage of the activity was in a soluble state; these features were reflected in the histochemical demonstration of acid phosphatase by an intense staining reaction which was judged to be unsatisfactory. The reverse held true for tissues fixed in aldehydes of the third group, using glutaraldehyde in the staining tests Evidence is presented suggesting that acid phosphatase staining results after formaldehyde fixation is dependent in part on the duration of fixation.
THE EFFECT OF NUTRITIONAL STATE ON PHOSPHORYLASE ACTIVITY IN THE LIVER: A HISTOCHEMICAL STUDY
Authors: FAGUNDE LUIZ A. , COHEN RICHARD B. 
Published: April 2011

Phosphorylase activity was studied in the hepatic lobule in the young male rat using a histochemical approach. In normal animals a diurnal variations in activity was observed characterized by low, irregularly distributed activity in the early morning with progressively increasing activity into the afternoon. In starved animals a more marked progressive increase in phosphorylase activity was demonstrated during the first 8 hr of starvation with increased activity seen throughout the lobule but accentuated in the periphery. After 12 hr of starvation there was a progressive decrease in centrilobular activity but peripheral activity remained undiminished until obout 48 hr of starvation, after which it also decreased. The increased activity in the afternoon in the normal a animal was attributed to the nocturnal feeding habits and resultant relatively poor nutritional state in the afternoon. Rise in phosphorylase activity in the early phase (4 hr) of starvation could be prevented by a single close of glucose by stomach tube. No effect of glucose was evident in intermediate term starvation until periods of 48-72 hr were reached, at which time a single dose of glucose by tube produced a marked increase in phosphorylase activity throughout the lobule within 4 hr of its administration. The basis for these findings and their possible significance is discussed.
CHOLINESTERASES AND MOTOR END-PLATES IN DEVELOPING DUCK SKELETAL MUSCLE
Authors: KHERA K. S. , LAHAM Q. N. 
Published: April 2011

End-plates in the thigh muscles of duck embryos were first recognized with myristoylcholine as substrate at the 19th day of incubation. Each appeared as a cholinesterase-positive dot surrounded by a small halo which rapidly increased in size during the 20th and 21st days. The endplates were usually oval in shape, averaging 33 µ x 25 µ with a subneural apparatus 5-12µ wide. The latter contained refringent lamellas arranged transversely in a palisade fashion. From the 21st day to the day of hatching (27-29 days) the number of end-plates progressively increased. After hatching, the myristoylcholine-reacting end-plates were difficult to locate. With acetylthiocholine as substrate, the embryonal end-plates were not demonstrable; however, the posthatched tissues showed numerous end-plates. The nerve trunks and nerve fibers gave a faintly positive myristoylcholine reactions in all stages after the 19th day of incubation. On the basis of the effects of eserine and diisopropyl fluorophosphate, the structures reacting with myristoylcholine and acetylthiocholine contained specific chohinesterase. The end-plates containing nonspecific cholinesterase also appeared on the 19th day of incubation and appeared to increase gradually in number until the 15th postembryonic day; thereafter they seemed to decrease.
HISTOCHEMICAL DEMONSTRATION OF THE SOLUBLE ENZYME, KETOSE REDUCTASE (SORBITOL DEHYDROGENASE), BY UTILIZING A NEW FIXING SOLUTION
Authors: JOHNSON ANNE B. 
Published: April 2011

Localization of the activity of the soluble enzyme, ketose reductase, was made possible by using a high-tonicity fixative containing formalin, ethanol, and tris (hydroxymethyl)aminomethane and by maintaining high tonicity in the incubating medium. Incubations could be performed at pH 8.8, an optimal pH for the conversion of sorbitol to fructose. False localization, probably resulting from diffusion of the first reactions product, reduced nicotinamide adenine dinucleotide, was minimized by slowing the reaction rate and using less tissue in the incubating medium. When this medium was tested with phenazine methosulfate, neither reduced nicotinamide adenine dinucleotide nor soluble dehydrogenase activity was detected. Ketose reductase activity was demonstrable throughout the epithelia of the seminal vesicle and the coagulating gland of the mouse. The activity was greater in the coagulating gland. It also was present in spermatozoa, central hepatic parenchymal cells, and renal proximal tubule. It was not present in the vas deferens epithelium nor ganglion cells. Reactivity in smooth muscle was believed to be artifact.
HISTOCHEMICAL DEMONSTRATION OF SOME OXIDIZED MACROMOLECULES WITH THIOCARBOHYDRAZIDE (TCH) OR THIOSEMICARBAZIDE (TSC) AND OSMIUM TETROXIDE
Authors: SELIGMAN ARNOLD M. , HANKER JACOB S. , WASSERKRUG HANNAH , DMOCHOWSKI HERMINE , KATZOFF LIONEL 
Published: April 2011

Thiocarbohydrazide (TCH) and thiosemicarbazide (TSC) have been introduced as osmiophilic reagents for demonstrating aldehyde-containing macromolecules, produced by periodic acid oxidation of tissue sections. Because of the remarkable properties of osmium black, excellent pigment qualities are provided for light microscopy and the advantages of an amorphous, nonvolatile, electron-opaque end product are available for electron microscopy. The present communication describes the method and gives the scope of the histochemical reaction. Thiocarbohydrazide is the more reactive of the two reagents and is capable of demonstrating a wider variety of oxidized macromolecules than does thiosemicarbazide. The periodic acid-thiocarbohydrazide-osmium tetroxide method (PATCO method) was compared with those using thiosemicarbazide (PATO method), periodic acid-Schiff (PAS method), and with the method involving 3-hydroxy-2-naphthoic acid hydrazide (NAH) followed by coupling with fast blue B (PA-NAH-FBB method). All four methods demonstrated certain active aldehydes equally well, such as those produced by periodic acid oxidation of glycogen, mucopolysaccharide of gut and mucoprotein of Descemet's membranes of the cornea. The stain with the PATO method was somewhat weaker than the other three methods for glycoprotein of the cuticular border of gut and of the brush border of kidney tubules, and colloid of the thyroid gland. The PATO method was even less effective in demonstrating mucoprotein of basement membranes in kidney and no reaction for collagen and reticulin was noted. The PATCO method was almost as effective with basement membranes in kidney as the PAS method, but was not equal to the PAS method in staining collagen and reticulin. Furthermore, the aldehyde produced on Feulgen hydrolysis of deoxyribonucleic acid (DNA) of cell nuclei was not demonstrable with either PATO or PATCO methods its contrast to positive results with the PAS and PA-NAH-FBB methods. This suggests that TSC and TCH have less reactive hydrazino groups than NAH. Results with the electron microscope will be published separately.
STUDIES IN HISTOCHEMISTRY: LXXXII. DETERMINATION OF VITAMIN A IN MICROGRAM SAMPLES OF TISSUE AND ITS QUANTITATIVE HISTOLOGICAL DISTRIBUTION IN THE RAT ADRENAL
Authors: NAKANE PAUL K. , GLICK DAVID 
Published: April 2011

A fluorometric procedure is presented for the determinations of vitamin A in small tissue samples, e. g., microtome sections, to enable correlation of analytical data with histology. Measurements may be made over the range, 3 x 10–12–6 x 10–10 moles of the vitamin (0.86-17 mµg of vitamin A alcohol), the quantities per 60 µl of solution used in the microcuvettes for fluorometry. Optimal conditions for certain of the analytical steps and for extraction of the vitamin from tissue were studied. Quantitative recoveries of vitamin added to tissue were obtained, and the reproducibility of the analysis of tissue was observed to have a standard deviation of 4%. In preliminary experiments, a mean vitamin A per rat adrenal was found to be 2.3 mµmoles, and the mean concentration 317 mµmoles/g, dry weight. A study of the quantitative histological distribution of the vitamin in the adrenal revealed greatest concentrations (4.7 x 10–12 moles/µg of proten-nitrogen) in the region where the glomerulosa and fasciculata join, and the concentration fell steadily through the fasciculata and reticularis to a low (0.9 x 10–12 moles/µg of protein-nitrogen) in the medulla.
A REVIEW OF EARLY CONCEPTS OF AMYLOID IN CONTEXT WITH CONTEMPORARY CHEMICAL LITERATURE FROM 1839 TO 1859
Authors: PUCHTLER HOLDE , SWEAT FAYE 
Published: April 2011

Early concepts of amyloid are reviewed in context with chemical literature from 1839-1859. Cellulose was first described in 1839. Treatment with sulfuric acid converted cellulose into a substance which was colored blue by iodine. This cellulose derivative was called amyloid. Virchow applied the chemical tests for starch and cellulose to human tissues. The substance in waxy degeneration reacted like immature cellulose. Virchow did not coin the term amyloid for this substance but suggested it as a compromise. Virchow restricted the term amyloid to the carbohydrate moiety and explicitly excluded the protein fraction which he had described earlier. Friedreich and Kekulé (1859) studied an evidently impure sample of amyloid and concluded that it did not contain polysaccharides. Since then the protein moiety has been studied extensively, but the carbohydrate fraction has been largely neglected. It appears possible that investigations of the carbohydrates at sites of amyloid formation would aid the understanding of the pathogenesis of amyloidosis2.
FINE STRUCTURAL LOCALIZATION OF ACID AND ALKALINE PHOSPHATASES IN CELLS OF RABBIT BLOOD AND BONE MARROW
Authors: WETZEL B. K. , SPICER S. S. , HORN R. G. 
Published: April 2011

In rabbit heterophils, acid phosphatase activity occurs in primary (azurophil) granules which predominate in early cells and persist in mature cells and in tertiary granules which are seen only in mature cells. Alkaline phosphatase activity occurs in secondary granules which appear in intermediate heterophils and later predominate in mature cells. Acid phosphatase activity in heterophil Golgi zones coincides developmentally with the genesis of primary and, later, tertiary granules, whereas alkaline phosphatase in the Golgi complex coincides with secondary granulogenesis. In developing eosinophils, acid phosphatase reaction product occurs in Golgi elements, rims the spherical precursors of angular, mature granules and appears inconsistently within mature granules. Basophil myelocytes show acid phosphatase in Golgi elements but not in specific granules. Additional acid phosphatase reactive structures include: granules of mononuclear cells; phagocytic vacuoles in macrophages; autophagic vacuoles in maturing erythroid cells; small dense granules of platelets; dense bodies in lipocytes; and Golgi elements of mononuclear cells, macrophages, nucleated red cells, megakaryocytes and lipocytes. Localized deposits were absent in control specimens except for enzyme-independent nuclear staining in alkaline phosphatase preparations.
THE ROLE OF HISTOCHEMICAL AND BIOCHEMICAL PREPARATION METHODS FOR THE DETECTION OF GLYCOGEN
Authors: LESKE R. , MAYERSBACH H. v. 
Published: April 2011

Block fixation for the histochemical demonstration does not safely preserve glycogen. Even when using "glycogen fixatives" and low temperatures the staining measured intensity does not correspond to the amount of glycogen measured chemically. Adequate preservation is obtained only with quenching. Lyo- and desmoglycogen fractions in biochemical assays are obtained when coarse tissue homogenates are extracted with TCA, while from fine homogenates the same glycogen amount can be extracted with TCA as with KOH digestion of the tissues. The histochemical demonstrability of glycogen in fixed tissue blocks does not depend on the presence of lyoglycogen.