bluuebunny/crossref_metadata_2025 huggingface dataset:
THE CONNECTIVE TISSUE HISTOCHEMISTRY OF NORMAL AND SOME PATHOLOGICAL SKIN
Authors: SAMS W. MITCHELL , SMITH J. GRAHAM , DAVIDSON EUGENE A.
Published: April 2011

A detailed histochemical study of the connective tissue of the skin has been presented. The stains used included hematoxylin and cosin, Mowry's colloidal iron and alcian blue for acid mucopolysaccharides, bromphenol blue for proteins, synthetic orcein and Verhoeff's for elastic tissue, and a combined Mowry's colloidal iron-orcein stain. Blocking procedures employed included methylation, demethylation, acetylation, deamination, and oxidation. Enzyme extractions were performed with testicular hyaluronidase.

Representative diseases of the following three types were studied: 1) those due to or influenced by the effects of solar radiation; 2) those associated with abnormal elastic fibers without particular relationship to the amount of actinic radiation; and, 3) those associated with large numbers of fibroblasts.

In chronically sun-damaged skin such as actinic elastosis, actinic keratosis, and basal cell epithelioma, markedly increased amounts of acid mucopolysaccharides were found in the areas of basophilic and orceinophilic material. Polymorphic light eruption and discoid and disseminate lupus erythematosus were found to have numerous acid mucopolysaccharide strands throughout the upper one-half of the dermis. Elastosis perforans serpiginosa displayed increased acid mucopolysaccharides associated with its orceinophilic fibers. Approximately 75% of the acid mucopolysaccharide in sun-damaged skin and elastosis perforans serpiginosa was removed by testicular hyaluronidase.

In pseudoxanthoma elasticum each curled elastic fiber was found in the center of a pool of acid mucopolysaccharide, as though, when it retracted, it brought its mucopolysaccharide sheath with it. This acid mucopolysaccharide, in contrast to that found in sun-damaged skin and elastosis perforans serpiginosa, is only slightly labile to testicular hyaluronidase.

Acid mucopolysaccharides were also found in large quantity wherever there were young fibroblasts as in dermatofibromas, neurofibromas, keloids, and fibroepithelial polyps, and is believed by the authors to represent de novo production by these cells rather than "mucinous degeneration." Hyaluronidase removed approximately 75% of the acid mucopolysaccharides in all conditions associated with young fibroblasts.

The usefulness of the combined Mowry-Haleorcein stain was stressed.

From the evidence presented, doubt was cast on the presently held concept that colloidal iron binding is by dissociated acid groups alone; iron may also be chelated by the intermolecular hydrogen bond of acid mucopolysaccharides.

Oxidation greatly increased the intensity of Mowry-Hale staining: bromine probably by direct bromination and formation of cysteic acid and permanganate probably by formation of carboxyl groups.
ADENOSINE TRIPHOSPHATASE: A NEUROHISTOCHEMICAL STUDY
Authors: NAIDOO D.
Published: April 2011

The location of adenosine triphosphatase in the brain has been studied in rapidly frozen-dried cerebral tissues of the Wistar rat.

It is found that adenosine triphosphatase is an almost exclusively nuclear enzyme.

Two tissue fractions of the cerebrum were separated, so that one sample was made up of vascular elements, and the other of neural elements. The two fractions were then studied for their adenosine triphosphatase activity, and compared with the histochemical findings.

The two tissue fractions were found not to differ in the absence of bivalent cations. When Ca++ were added to the cerebral vascular suspension, ATPase activity was increased approximately 15 times, and only 3 times in the presence of Mg++. Conversely, the addition of Mg++ increased the ATPase activity of the neural fraction 200%; whereas, Ca++ was responsible for a 60% increase.

This fact was detectable microscopically when Ca++ was found to intensify vascular nuclear staining, and Mg++ to increase the neuronal and glial nuclear staining.

The results, histochemical and biochemical, are mutually confirmatory.
Randomized Controlled Studies and Alternative Designs in Outcome Studies
Authors: Shadish William R.
Published: April 2011

This article reviews several decades of the author’s meta-analytic and experimental research on the conditions under which nonrandomized experiments can approximate the results from randomized experiments (REs). Several studies make clear that we can expect accurate effect estimates from the regression discontinuity design, though its statistical power is lower, it estimates a different parameter than the RE, and its analysis is considerably more complex. For other nonrandomized designs, the picture is more complex. They may yield accurate estimates if they are prospectively designed to include comprehensive and reliable measurement of the process by which participants selected into conditions, if they use large sample sizes, and if they carefully select control groups that are from the same location and with the same substantive characteristics. By contrast, we have little good reason to think that nonrandomized experiments using archival data without comprehensive selection measures are likely to yield accurate effect estimates.
Delirium in Palliative Care Patients: Focus on Pharmacotherapy
Authors: Zimmerman Kristin , Rudolph James , Salow Marci , Skarf L. Michal
Published: April 2011

Patients receiving palliative care often possess multiple risk factors and predisposing conditions for delirium. The impact of delirium on patient care in this population may also be far-reaching: affecting not only quality of remaining life but the dying process experienced by patients, caregivers, and the medical team as well. As palliative care focuses on comfort and symptom management, the approach to assessment and subsequent treatment of delirium in palliative care patients may prove difficult for providers to navigate. This article summarizes the multifactorial nature, numerous predisposing medical risk factors, neuropsychiatric adverse effects of palliative medications, pharmacokinetic changes, and challenges complicating delirium assessment and provides a systematic framework for assessment. The benefits, risks, and patient-specific considerations for treatment selection are also discussed.
Nipple Involvement in Breast Cancer: Retrospective Analysis of 2323 Consecutive Mastectomy Specimens
Authors: Weidong Li  , Shuling Wang  , Xiaojing Guo  , Ronggang Lang  , Yu Fan  , Feng Gu  , Xinmin Zhang  , Zhengmao Zhu  , Yun Niu  , Li Fu
Published: April 2011

Breast cancer surgical options now include nipple-sparing mastectomy (NSM), but there has been much controversy regarding the oncologic safety of the preserved nipple. This study evaluates frequency and patterns of occult nipple involvement in a large contemporary cohort of patients, aiming to improve patient selection for NSM. A total of 2323 consecutive mastectomy specimens with grossly unremarkable nipples were evaluated by sagittal sections through the entire nipple and subareolar tissue. Sixteen different clinical and tumor parameters were examined to predict cancerous nipple involvement. Nipple involvement was noted in 331 of 2323 (14.2%) mastectomy specimens. Occult nipple involvement rate was 10.7% (248 cases). Occult nipple involvement usually occurs as ductal carcinoma in situ. In univariate analysis, patient age, tumor size, tumor-to-nipple distance, tumor central location, tumor type, lymph node status, lymphatic vascular invasion, histologic grade, estrogen receptor, progesterone receptor, human epidermal growth factor receptor 2 (HER-2) amplification, and multicentric or multifocal tumor were associated with positive nipple involvement. By multivariate logistic regression analysis with the entire selection process, tumor size, tumor-to-nipple distance, central location, lymph node status, lymphatic vascular invasion, HER-2 amplification, and multicentric or multifocal tumor were shown to be associated with nipple involvement by carcinoma. Nearly 90% women undergoing mastectomy did not have occult nipple involvement. NSM may be a viable option in appropriate patient selection and setting. NAC preservation would be appropriate in HER-2 negative, axillary lymph node, and lymphatic vascular negative patients with small, solitary tumors located on the periphery of the breast.
Epstein–Barr Virus–Associated Adrenal Smooth Muscle Tumors and Disseminated Diffuse Large B-Cell Lymphoma in a Child With Common Variable Immunodeficiency
Authors: Petrilli Giulia , Lorenzi Luisa , Paracchini Roberto , Ubiali Alessandro , Schumacher Richard Fabian , Cabassa Paolo , Facchetti Fabio
Published: April 2011

This article reports the clinical and the histological features in a 7-year-old girl affected by common variable immunodeficiency (CVID) who developed multiple Epstein–Barr virus–associated tumors, represented by bilateral adrenal smooth muscle tumors (EBV-SMT) and multifocal diffuse large B-cell lymphoma. The EBV-SMTs showed features compatible with a benign or at least a low-malignant potential neoplasm. A peculiar feature observed in both EBV-SMTs was the occurrence of numerous lymphocytes intermingled with the spindle cells, which consisted of CD3+ CD5+ T-cells, with a predominant cytotoxic CD8+ component. Interestingly, EBV status differed in the neoplasms, since the EBV-SMTs were negative for LMP1 and positive for EBER, whereas the B-cell lymphoma expressed both EBV markers. Furthermore, EBV-LMP1 deletion was positive only in the EBV-SMTs, thus indicating that these tumors were the consequence of 2 distinct, EBV-dependent transformations. Similarly, lymphocyte clonality assay also showed different clonal bands in different sites (skin and nasal cavity), suggesting the development of intratumoral mutations. Finally, the authors review all 127 previously reported EBV-SMT, with discussion of their clinical and pathological features.
The Potential Effects of IGF-1 and GH on Patients With Chronic Heart Failure
Authors: Perkel David , Naghi Jesse , Agarwal Megha , Morrissey Ryan P. , Phan Anita , Willix Robert D. , Schwarz Ernst R.
Published: April 2011

Heart failure (HF) is an important health concern with almost a quarter million deaths each year despite advances in medical therapy. Improvement of cardiac function has been shown to reduce morbidity and mortality in patients with HF. There has been recent interest in the growth hormone (GH) / insulin-like growth factor (IGF) pathway as a potential therapeutic target for patients with HF. Insulin-like growth factor 1 has been shown to augment cardiac function ex vivo and in animals. It was hypothesized that IGF-1/IGF-binding protein 3 levels might be able to provide prognostic benefits in patients with heart disease. Initial observational studies have shown significant benefits from GH supplementation including improved ejection fraction, increased exercise tolerance, and decreased New York Heart Association functional class. These results, however, were not replicated in randomized, controlled trials. Patients with advanced stages of HF might develop cachexia associated with a state of significant GH resistance. The lack of response to GH supplementation may be secondary to a deficiency in IGF-1, the effector hormone. Hypothetically, this group of patients could benefit from direct IGF-1 supplementation. Combined therapy with GH and IGF-1 is appealing; however, future trials in patients with advanced HF are warranted to prove this concept.
POLYSACCHARIDES OF CONNECTIVE TISSUE
Authors: DORFMAN ALBERT
Published: April 2011

An attempt has been made to review the current status of knowledge of the chemistry and metabolism of the acid mucopolysaccharides of connective tissues. It has been pointed out that complex polysaccharides play a prominent role in the physiology of cell surface as well as the intracellular matrix. In mammalian tissues, connective tissues contain a variety of acid mucopolysaccharides with varying structure and biological activity. It is probable that their localization is correlated with their functional capacities.

Rapidly progressing understanding of biosynthetic mechanisms indicates the pathways of biosynthesis of some members of this class of compounds although many important questions remain unsolved. It is apparent that the biosynthesis of these compounds is under control of a number of factors, most strikingly the endocrine agents.
TRANSFERENCE OF CIRCULATING LABELED SERUM PROTEINS TO THE FOLLICLE OF THE RAT OVARY
Authors: MANCINI R. E. , VILAR O. , HEINRICH J. J. , DAVIDSON O. W. , ALVAREZ B.
Published: April 2011

Rat whole serum, albumin, globulins and fibrinogen fractions were labeled with fluorescent dyes (sulforhodamine B, CI No. 45100, and fluorescein isothiocyanate); albumin was also tagged with radioactive iodine (I131). Labeled serum fractions were intravenously injected in adult female albino rats and their decay in the circulation and histological distribution in the ovaries was studied. As a complement histoimmunological techniques using labeled fluorescent antibodies against rat serum fractions were also applied to the ovary sections of intact rats as well as of rats previously injected with unlabeled serum fractions. It was observed : 1) Labeled whole serum, albumin and globulins decay in the circulation with a half-life value of 2.8 days for the first, 2.6 days for the second and 3.1 days for the last. 2) Parallel with the fast component of this time decline curve, serum fractions diffuse extravascularly in the ovary and appear in the zona pellucida and cytoplasm of the oocyte in the growing follicles, via thecal capillaries, intercellular spaces of granulosa layer and antrum. 3) Histoimmunological techniques suggest that also endogenous circulating serum fractions may be transmitted to the follicles following the same route.
THE DEMONSTRATION OF ALKALINE PHOSPHATASE ACTIVITY IN FROZEN-DRIED MOUSE GUT IN THE ELECTRON MICROSCOPE
Authors: CHASE WILLIAM H.
Published: April 2011

Fragments of mouse gut mucosa were frozen-dried and embedded in prepolymerized methacrylate. Serial sections were examined in the light and electron microscope after staining with Gomori's method for alkaline phosphatase. The tissue showed high enzyme activity. In the electron microscope deposits of calcium phosphate were seen in the brush border and the Golgi area of epithelial cells, and in large and small apical granules.
ENZYMATIC ACTIVITY OF PANCREATIC ISLETS AND ACINI IN NORMAL AND TOLBUTAMIDE-TREATED RATS
Authors: KISSANE JOHN M. , BROLIN SVEN E.
Published: April 2011

By quantitative microchemical techniques, four enzymes—malic, lactic, and glucose-6-phosphate dehydrogenases, and glutamic-oxalacetic transaminase-were measured in pancreatic islets and acini from rats 25 hours after an injection of tolbutamide (150 mg/kg) and compared with controls from animals receiving either saline or no injections. Malic and glucose-6-phosphate dehydrogenase and transaminase were all three to four times as active in islets as in acini, while islet lactic dehydrogenase presented only one-third to one-fourth that activity in acini. No significant differences were observed among tolbutamide, saline or non-injected animals. The pancreas of the rat thus presents a distinctive enzymatic pattern susceptible to quantitative studies.
STUDY OF TISSUE MUCINS USING THE FLUORESCENT ANTIBODY TECHNIQUE: II. THE PREPARATION AND SPECIFICITY OF HUMAN SUBMAXILLARY GLAND MUCIN ANTIBODY
Authors: KENT SIDNEY P.
Published: April 2011

Mucin was extracted from human submaxillary glands and purified. Antibodies to the mucin were produced in rabbits and labelled with fluorescein isothiocyanate. The labelled antibody was used to demonstrate mucin in paraffin embedded tissue sections of human submaxillary glands. The antibody solution reacted with the mucin in human submaxillary glands but not with the submaxillary glands of other species. Cross reactions were noted with epithelial mucins in human sublingual glands, lingual glands, submucosal glands of the trachea, gallbladder, pancreatic ducts, Bartholin glands and cervical glands. The organ specificity of the antibody solution was improved by absorbing the antibody solution with sections of Bartholin glands i.e. the cross reactions noted with the Bartholin glands was eliminated but the antibody solutions continued to react with human submaxillary glands. The relation of the antibody to human submaxillary mucins to blood group factors is discussed. Possible ways of obtaining antibodies with a high degree of organ specificity are noted.
THE INFLUENCE OF TECHNIQUE AND HORMONES ON HISTOCHEMICAL DEMONSTRATION OF ENZYMES IN INTESTINAL EPITHELIUM
Authors: LIU HSI-YEN , BAKER BURTON L.
Published: April 2011

The influence of technical variations and of hypophysectomy and adrenalectomy on the demonstration of enzymes in the jejunal epithelium was studied. The application to fresh frozen sections of procedures for cytochrome c oxidase, succinic dehydrogenase, diphosphopyridine nucleotide diaphorase and lactic dehydrogenase failed to reveal a precise mitochondrial localization, basing the conclusion on comparison with mitochondria as demonstrated by staining of Regaud-fixed mucosa with the Altmann-Masson procedure. Fixation in cold formalin prior to freezing and sectioning permitted a more satisfactory localization of succinic dehydrogenase in mitochondria.

Hypophysectomy or adrenalectomy resulted in a reduction in number, size, staining and orderly arrangement of mitochondria in the villous epithelium as revealed in cytological preparations and those fixed and stained for succinic dehydrogenase. Alkaline phosphatase activity was also depressed. The intensification of cytoplasmic staining for lactic dehydrogenase, diphosphopyridine nucleotide diaphorase and succinic dehydrogenase in fresh frozen tissue probably represents concentration of formazan by reduction in cell volume and not increased enzyme activities.
A BIOCHEMICAL AND HISTOCHEMICAL STUDY OF β-d-GLUCURONIDASE ACTIVITY IN HAMSTER LIVER AND KIDNEY
Authors: MANNING J. P. , CAVAZOS L. F. , FEAGANS W. M. , MOSS R.
Published: April 2011

β-d-Glucuronidase activity was studied histochemically in male hamster liver and kidney under varied conditions of nonfixation and fixation. Results were evaluated according to degrees of reaction, localization and diffusion. The reference point for total enzyme activity in these organs was determined biochemically in similarly nonfixed and fixed tissues. Consistent histochemical and biochemical results were obtained on nonfixed tissues and with material fixed in formalin-gum-sucrose. Organs fixed in formol-chloral hydrate and neutral buffered formalin were histochemically and biochemically negative for glucuronidase activity. Levels of β-d-glucuronidase in hamster liver and kidneys are low in comparison to known activity in the rat. Apparently, in the hamster, this enzyme is not present in sufficient quantity to give a positive reaction following partial inactivation by formol-chloral hydrate or neutral buffered formalin fixations. It is suggested that studies on tissues low in β-d-glucuronidase activity should incorporate both nonfixed and fixed tissues.
A HISTOCHEMICAL STUDY OF THE DERMO-EPIDERMAL MEMBRANE IN COWHIDE
Authors: BOTAN E. A. , CHOUINARD A. E.
Published: April 2011

The dermo-epidermal membrane was demonstrated to have fibrous elements, a few scattered cellular elements, and amorphous elements. The fiber elements were reticular fibers (the principal fiber) and elastin. A few scattered fibroblasts were demonstrated close to the area of the corium minor. The amorphous material is suggested to be mucoprotein and/or glycoprotein.
ELECTRO- AND IMMUNOELECTROPHORETIC PATTERNS IN THE LARVAL SALIVARY SECRETION OF CHIRONOMUS THUMMI
Authors: KATO K.-I. , PERKOWSKA E. , SIRLIN J. L.
Published: April 2011

The salivary secretion of chironomid larvae was studied electro- and immunoelectrophoretically.

The secretion could be resolved into 14 nigrosin-positive (protein) bands and 10 metachromatic (mucopolysaccharide) bands in the pherograms. Much less stainability was found in some of these bands in the pherograms of the gland cell components. The polysaccharide bands roughly corresponded with an equal number of protein bands at the same positions in the pherogram.

An antiserum against a cell extract of whole larvae (excluding the salivary glands) yielded 6 groups of precipitation crescents on the pherogram of the cell extract. The same antiserum which had been previously absorbed with the salivary secretion yields a reduced number of crescents, specially on the anodic side of the pherogram.
A MODEL DARK ROOM UNIT FOR RADIOAUTOGRAPHY
Authors: KOPRIWA BEATRIX MARKUS
Published: April 2011

The layout and equipment of a dark room unit for radioautography are described. In addition to an entrance hall and a small corridor, the unit includes two main parts: a temperature- and humidity-controlled coating room, where the histological sections are coated with photographic emulsion and dried; and a temperature-controlled processing room, where the preparations are develped and fixed after exposure.
CHARACTERISTICS OF THE ESTERASES OF HUMAN CELLS GROWN IN VITRO
Authors: KOMMA DONALD J.
Published: April 2011

When extracts of cultured human cells are subjected to starch-gel electrophoresis, as many as 17 bands of esterase activity can be observed. In addition to their different migratory characteristics in an electric field, the various bands can be shown to differ in their substrate preferences, their stability during pronlonged storage and during heating, and their interactions with inhibitors and activators. The abundance of types of esterase in these undifferentiated cells would seem to indicate a very important role for esterase reactions in cellular function.
DEHYDROGENASES IN DEVELOPING TEETH OF RATS
Authors: FULLMER HAROLD M.
Published: April 2011

Odontogenesis was studied in unfixed cryostatcut sections of 5-day old rats with histochemical methods for the demonstration of succinic, malic, isocitric, glutamic, lactic, β-hydroxybutyrate, α-glycerophosphate, glucose-6-phosphate and 6-phosphogluconate dehydrogenases and DPN and TPN diaphorases. Except for weak glucose-6-phosphate and 6-phosphogluconate dehydrogenases, odontogenic tissues (particularly ameloblasts, odontoblasts and stratum intermedium cells) exhibited moderate activity indicating a broad metabolic behaviour of these cells.

Cells of the stratum intermedium and the stellate reticulum display an enzymatic activity parallel to ameloblasts which suggests that they participate actively in amelogenesis.

Tetranitro-BT and Nitro BT appear to give definitive localization of dehydrogenase activity.
A HISTOCHEMICAL STUDY OF HYDROLYTIC ENZYMES IN MUSCLE FIBERS OF VARIOUS ANIMALS
Authors: OGATA TAKURO , MORI MASAHIKO
Published: April 2011

A histochemical study of hydrolytic enzymes was carried out by azo coupling methods. Alkaline phosphatase, acid phosphatase, esterase, aminopeptidase, β-glucuronidase, β-galactosidase, sulfatase and phosphorylase were histochemically demonstrated in muscles of various kinds of animals. Mammalian, bird, reptile and amphibian muscles contained esterase but no other enzymes, and a difference was observed in esterase activity in red, white and medium muscle fibers. Muscle fibers in mammals and birds showed the reciprocal proportion between phosphorylase and oxidative enzymes. Fish white muscle fibers showed a strong reaction of esterase, while the red muscle fiber a weak reaction. In the fish white muscle showed a slightly positive reaction of alkaline and acid phosphatase; aminopeptidase, β-glucuronidase and β-galaetosidase were slight positive in white fiber. Invertebrate muscles showed hydrolytic enzymes. These findings suggest that there is a difference in the metabolism in the muscle depending upon each species.
APPLICATION OF NAPHTHOL YELLOW S CYTOPHOTOMETRY IN DEOXYRIBONUCLEIC ACID AND PROTEIN DETERMINATIONS ON LARVAL MATERIAL OF D. MELANOGASTER
Authors: WELCH ROBERT M. , RESCH KATHLEEN
Published: April 2011

A modification of the Deitch "standard" naphthol yellow S technique is described and validated for use in deoxyribonucleic acid and protein determinations in drosophila larval material. By the use of this technique in conjunction with cytophotometry relative amounts of protein have been determined in 436 salivary gland nuclei from 37 individuals of wild-type, lgl lethal, and experimentally treated larvae of D. melanogaster and in 294 nuclei from 44 individuals by photometric interferometry. Altogether, 8 comparisons were made by comparing the ratio between two kinds of material obtained by naphthol yellow S cytophotometry with that obtained on the same kind of material by photometric interferometry. In 6 of the first 7 comparisons the cytophotometric ratio could be reconciled with the interferometric ratio by an adjustment well within statistical probability. In the seventh comparison, the ratios could not be reconciled by any adjustment within statistical probability but could be so reconciled when repeated on additional material of the same kind. Inconsistency of naphthol yellow results between the original and repeat comparisons indicated the probability of cytophotometric error due to nonproportionality of staining in the material used. Extensive additional tests revealed no further instance of inconsistency and implicated the material rather than the method in the one aberrancy discovered. It is concluded that the cytophotometric and interferometric data are in mutual support and indicate that either technique is a reliable method for protein determination; but that the naphthol yellow S technique may result in an occasional aberrancy in drosophila material that must be guarded against. Advantages and disadvantages of each technique are discussed.
VARIATIONS IN THE ELECTROPHORETICALLY SEPARATED ACID PHOSPHATASES OF TETRAHYMENA
Authors: ALLEN SALLY LYMAN , MISCH MARGARET SEGUR , MORRISON BARBARA M.
Published: April 2011

The acid phosphatases of variety 1 of Tetrahymena pyriformis can be separated into 17 zones by electrophoresis in starch gels of pH 7.5. All of these acid phosphatases have an optimal pH of about 5.0 and are inhibited by 10 mM sodium fluoride on d-tartaric acid. With one exception, all hydrolyze sodium α-naphthyl acid phosphate. Differences between the acid phosphatases are observed in their ability to hydrolyze other substrates using either the coupling technique or a modification of the Gomori-lead method. Inhibition of 2 of the acid phosphatases occurs in the presence of 1 mM of Mn++ or Zn++; 0.1 mM of p-chloromercurobenzoic acid inhibits these and others. Variations between the acid phosphatases were observed under different growth conditions and in their distribution in various cell fractions. A major variation in the acid phosphatases that is under genetic control occurs in extracts of different genotypes.

The results suggest that the electrophoretically separated acid phosphatases are a family of enzymes that vary in their degree of relationship. Some are different enzymes. Others are more closely related and represent variations of a single enzyme; i.e., mutant forms produced by different alleles, hybrid forms produced by interaction of alleles, and isozymes produced by a single gene.

Most of the acid phosphatases are probably associated with lysosomes, although one appears to be associated with microsomes. This acid phosphatase is much more active in synthetically grown cells than in cells grown in proteose-peptone or in bacterized medium. It does not hydrolyze sodium β-glycerophosphate.
HISTOCHEMICAL EVIDENCE OF PROTEOLYSIS IN BONE: THE INFLUENCE OF PARATHORMONE
Authors: BÉLANGER LEONARD F. , MIGICOVSKY B. B.
Published: April 2011

Proteolytic activity has been revealed at the level of the large osteocytes of young chick and rat bone by digestion of gelatin in a photographic emulsion substrate.

The reaction was enhanced by pretreatment of the animals with parathyroid extract.

The reaction was inhibited by addition to the incubating medium of sodium fluoride or potassium ferrocyanide. It was also inhibited by fixation in ethanol-formaldehyde acetic acid.

The present results seem to confirm and explain further the resorptive potential of the mature osteocytes.
REGIONAL UTILIZATION OF LEUCINE-H3 BY NORMAL RAT BRAIN: MICRODENSITOMETRIC EVALUATION OF AUTORADIOGRAMS
Authors: ALTMAN JOSEPH
Published: April 2011

Fine-resolution autoradiography was used to study the regional uptake of intraperitoneally administered leucine-H3 in the rat brain. Microdensitometric measurements were made of the differential rates of autoradiographic blackening over different brain regions, and low- and high-power photomicrographs were prepared of some representative sections of the neuraxis. Evidence of satisfactory consistency in regional density scores was obtained by comparing the grain density of several bilateral structures on the right and left side in single sections, and of homologous structures in different sections. It was concluded that fine-resolution autoradiography is a suitable technique for studying alterations in protein metabolism of homologous brain structures as a consequence of pathological changes in the brain and possible shifts due to physiological and behavioral manipulation of experimental animals.
NUCLEOSIDE PHOSPHATASE ACTIVITY IN MEMBRANOUS FINE STRUCTURES OF NEURONS AND GLIA
Authors: TORACK RICHARD M. , BARRNETT RUSSELL J.
Published: April 2011

The reaction product resulting from the hydrolysis of various nucleoside phosphate esters in the Wachstein-Meisel medium at a neutral pH has been localized by means of electron microscopy to various glial and neuronal membranous fine structures. Enzyme activity following the use of adenosine triphosphate as a substrate was localized to the interspace between the plasma membranes of neurons and glial dendrites adjacent to the neuronal cell body, the proximal axon and synaptic terminals. The relationship of this enzymatic activity to the transfer of cations, especially sodium and potassium, as well as to the synthesis of acetylcholine at synapses is discussed. A reaction product was also found in the Golgi apparatus of neurons and although it was most prominent following the use of inosine diphosphate as a substrate, it was also noted with other nucleoside phosphates. In addition, enzymatic activity was noted on the plasma membrane of oligodendroglia, particularly when adenosine triphosphate and cytidine triphosphate were used as substrates.
THE HISTOCHEMICAL LOCALIZATION OF HISTIDINE IN THE HUMAN EPIDERMIS AND ITS RELATIONSHIP TO ZINC BINDING
Authors: REAVEN EVE P. , CON ALVIN J.
Published: April 2011

A concentration of histidine in the transitional zone of the human epidermis has been identified by the diazonium coupling reaction. It has been concluded that histidine may be largely responsible for the selective in vitro zinc-binding property of the transitional zone of keratin formation in the epidermis.
HISTOCHEMICAL AND QUANTITATIVE CHANGES IN GLYCOGEN AND PHOSPHORYLASE DURING DISUSE ATROPHY OF THE PIGEON PECTORALIS
Authors: VALLYATHAN N. V. , CHERIAN K. M. , GEORGE J. C.
Published: April 2011

Histochemical and quantitative biochemical changes in glycogen content and phosphorylase activity of pigeon breast muscle were studied during development of disuse atrophy from one day to sixty days. Of the two types of muscle fibers present in the pigeon breast muscle the large white glycogen-loaded fibers, which are well adapted for an anaerobic metabolism, were found to lose their glycogen store and phosphorylase activity during the early days of atrophy. On the other hand, the smaller, red, fat-loaded fibers, which are well adapted for an aerobic metabolism, were found to acquire more glycogen and phosphorylase activity during the same period. The staining reaction of these fibers for phosphorylase thus, as with biochemically different branching enzymes, presented a mosaic. However, a few small fibers showed no increase in glycogen content or phosphorylase activity.

The histochemical observations also revealed that the majority of the large white fibers were reduced in size, filled with fat, and separated by connective tissue during the later stages of atrophy. Fibers of this sort disappeared or were completely engulfed by connective tissue. Very few small fibers changed in this way. In certain regions an entire fasciculus containing both types of fiber was found to be replaced by fat and connective tissue, leaving a few narrow red fibers intact. There was no phosphorylase activity in the connective tissue.

The quantitative studies showed a decrease in glycogen and phosphorylase levels in the muscle early in disuse. After 60 days of atrophy the phosphorylase level recovered somewhat but the glycogen level remained low.
NUCLEOSIDE PHOSPHATASE AND THIAMINE PYROPHOSPHATASE ACTIVITY OF RABBIT GOLGI APPARATUS
Authors: LAZARUS SYDNEY S. , WALLACE BARBARA J.
Published: April 2011

The Golgi associated nucleoside phosphatase which is demonstrable in rabbit pancreatic islet cells at alkaline pH using briefly postfixed cryostat sections and a modified Padykula-Herman incubation solution rapidly hydrolyzes uridine triphosphate (UTP) as well as uridine (UDP) and inosine diphosphate (IDP), but not thiamine pyrophosphate (TPP). Staining in the Golgi apparatus of islet cells was markedly attenuated when a lead incubation solution was employed at pH 7.2 with these same nucleoside-phosphates as substrates. The enzyme was not demonstrable in sections cut from tissue blocks fixed for 24 hours in formalin. This enzyme was distributed in the Golgi apparatus of some cellular elements of many other rabbit tissues. In rabbit cortical neurons a different Golgi enzyme was demonstrated which at alkaline pH hydrolyzed TPP but not UTP, IDP or UDP. This enzyme was also demonstrable with the lead method at pH 7.2 and under these conditions hydrolyzed IDP and UDP as well as TPP, but not UTP. The neuronal enzymatic activity was much less formalin-sensitive than that in pancreatic islet Golgi apparatus surviving in 24-hr formalin-fixed tissue blocks. This latter enzyme could be demonstrated in addition to brain only in the Golgi apparatus of hypophysis and epididymis. Differences in substrate specificity and of response to adjuvants as well as the differences in distribution confirm that these are two distinct enzymes.
THE HISTOCHEMICAL DEMONSTRATION OF ADENOSINE TRIPHOSPHATASE ACTIVITY IN MYOCARDIUM
Authors: NILES N. R. , CHAYEN J. , CUNNINGHAM G. J. , BITENSKY LUCILLE
Published: April 2011

Adenosine triphosphatase has been demonstrated histochemically in rat and human myocardium. To obtain its precise localization in discrete bands, apparently corresponding to the concentration of myosin, it was necessary to modify the existing technique to obtain better preservation of unfixed tissue and maximal enzymatic activity. Thus it was necessary to increase the concentration of calcium and to effect the reaction at pH 9.4 after treatment with 2:4-dinitrophenol. The specificity of the reaction was shown by these factors, by testing with phosphate esters other than adenosine triphosphate, and by the inhibitory effect of magnesium.
THE MECHANISM OF THE SCHIFF REACTION AS STUDIED WITH HISTOCHEMICAL MODEL SYSTEMS
Authors: HARDONK M. J. , VAN DUIJN P.
Published: April 2011

This report describes results obtained by staining several histochemical model systems and tissue sections by a number of Schiff-type reactions. From the results it may be concluded that the generally accepted hypothesis put forward by Wieland and Scheuing (14) about the reaction between aldehydes and Schiff's reagent must be rejected Agreement with the alkylsulfonic acid theory introduced by Rumpf (12) and Hörmann et al. (7) was found.
CYTOCHEMICAL DEMONSTRATION OF ACID PHOSPHATASE IN HEMATOPOIETIC CELLS IN HEALTH AND IN VARIOUS HEMATOLOGICAL DISORDERS USING AZO DYE TECHNIQUES
Authors: KAPLOW L. S. , BURSTONE M. S.
Published: April 2011

Acid phosphatase activity in the formed elements of blood and bone marrow has been demonstrated using unfixed smears and azo dye coupling techniques with naphthol AS-MX or AS-TR phosphates as substrates. A similar staining pattern was also obtained using a new substrate, hydroxycarbazole phosphate. When this compound was used, hydrolysis was more rapid and good staining was obtained using smears fixed at room temperature (25°C) in 60% acetone buffered to pH 4.2-4.5 with citrate. Such preparations showed good cytological detail; however, the azo dye is less chromogenic than that formed with the substituted naphthol phosphates and nonspecific staining due to free hydroxycarbazole was observed. The use of ethylene diamine tetraacetic acid (EDTA) as an anticoagulant did not interfere with the reaction. Moderate reduction in staining intensity was observed using smears stored at room temperature longer than 24 hours prior to incubation.

Dye precipitation was observed in most cells and was strongest in bone marrow osteoclasts and phagocytic reticulum cells. Immature myeloid cells showed moderately strong activity and this was particularly evident in peripheral blood smears of some patients with untreated chronic myelogenous leukemia. Increased staining was also observed in the plasma cells of 2 patients with untreated multiple myeloma. No consistent deviation from the normal was observed in the remaining disorders studied.
SOME HISTOCHEMICAL REACTIONS FOR LIPIDS IN AVIAN NERVOUS TISSUES. I. CENTRAL NERVOUS TISSUES
Authors: SINGH RAMESHWAR
Published: April 2011

The histochemical localization of lipids, as obtained by the application of the acid hematein test of Baker (1946), the controlled chromation method of Elftman (1958), the copper phthalocyanin method of Pearse (1955) and the phosphomolybdic acid method of Landing, Uzman and Whipple (1952), in avian central nervous tissues has been given. For the localization of intraneuronal phospholipids, good results were obtained only with the controlled chromation method of Elftman (1958). The intraneuronal phospholipid appears to be present in the form of granular bodies almost uniformly distributed in the cell perikaryon. The other methods given above failed to demonstrate intraneuronal lipids. However, all these work for the phospholipids of myelin. The results obtained have been discussed and compared with those available in the literature on neurohistochemistry in general.
TETRACYCLINE FLUORESCENCE IN MOUSE GASTRIC CARCINOMA
Authors: YABE Y. , WARREN I. A. , BERK J. E. , KOBERNICK S. D.
Published: April 2011

Tetracycline was administered to a strain of C57BL mouse harboring an experimental and transmissible tumor, gastric adenocarcinoma No. 328. Using fluorescent and phase microscopy, the fluorescence induced by the drug was localized to the cytoplasm of the tumor cells and to the cytoplasm of normal gastric mucosal and liver parenchymal cells. None was observed in the stroma. The fluorescence appeared rapidly but was retained by the tumor and normal cells for only 48 to 72 hours in contrast to the long lasting fluorescence present in bone and teeth, and that observed in some patients with carcinoma.
ELECTROPHORETIC EVIDENCE FOR ISOZYMES OF ARYLAMIDASE ("AMINOPEPTIDASE") IN THE RAT. I. RENAL, SERUM AND URINARY ENZYMES SPLITTING dl-ALANYL-2-NAPHTHYLAMIDE AND l-LEUCYL-2-NAPHTHYLAMIDE
Authors: MONIS BENITO
Published: April 2011

Bilateral nephrectomy elicited no changes of serum blood levels of arylamidase assayed with two chromogenic substrates (l-leucyl-2-naphthylamide and dl-alanyl-2-naphthylamide) two days after the operation. Since rat urine contains similar enzymes it was postulated that the urinary enzyme is of renal origin and distinct from serum arylamidase. For this purpose, electrophoretic studies were undertaken. Starch block (for quantitative determinations) and starch gel for zymograms of arylamidase were used. It was shown that both procedures demonstrated the identity of urinary and renal arylamidase, which was distinct from the serum enzyme. The renal and urinary enzyme showed two distinct isozymes: one remaining at the origin and the other migrating somewhat less than the isozyme in serum. It is postulated that the isozyme remaining at the origin corresponds to a membrane-bound form, whereas the electrophoretically-mobile isozyme is found in the cellular supernatant fraction. The arylamidase from serum was represented by a single enzyme which migrated farthest towards the anode.

Histochemical procedures for the demonstration of arylamidase activity in tissues at the light microscopic level permit the localization of enzyme(s) that can hydrolyze synthetic chromogenic naphthylamides containing l-leucyl and dl-alanyl groups (1).

Rat kidneys have a high concentration of histochemically demonstrable arylamidase in the proximal convoluted tubules (2). Blood levels of arylamidase in the normal adult rat vary within a narrow range (3). The observation of arylamidase activity in rat urine raised several questions that led to the studies which are the basis of the present report.

The relationship of serum and urinary arylamidase was the starting point of this investigation. It was speculated that if the urinary enzyme had its origin in the serum, bilateral nephrectomy should alter blood levels of the enzyme. If no such changes occurred, this would suggest that the urinary enzyme was released from the proximal convoluted tubule. In fact, total renal ablation led to no significant variation of serum arylamidase activity.

It was therefore postulated that urinary arylamidase originated in the kidneys and that both are distinct from serum arylamidase. To test this hypothesis, zone electrophoretic studies were undertaken using two different supporting media. It will be shown that distinct molecular forms or isozymes of arylamidase can be separated from rat tissues and fluids by differential electrophoretic mobility. The electrophoretic identity of urinary and renal arylamidase, both of which are distinct from the serum arylamidase, is demonstrated in this study.
A SILVER STAIN FOR DEOXYRIBONUCLEIC ACID
Authors: KORSON ROY
Published: April 2011

By hydrolyzing tissue sections with 1 M citric acid for 30 minutes at 60°C and subsequently treating the sections with methenamine silver for 60 minutes at 60°C (5% silver nitrate in 3% aqueous methenamine), a stain has been developed which is extremely intense and specific for nuclear deoxyribonucleic acid. This has been tested in a number of ways including deoxyribonuclease, trichloroacetic and perchloric acid extraction. The reaction has been successful with fresh frozen cryostat sections as well as with paraffin-embedded material. The stain is likewise satisfactory with tissue fixed in neutral formalin, Carnoy's solution, or formalin-phosphate-sucrose mixture. Because the silver precipitate is precisely localized it may be useful in visualizing chromatin with the electron microscope.
ON THE BINDING OF DIRECT COTTON DYES BY AMYLOID
Authors: PUCHTLER HOLDE , SWEAT FAYE , KUHNS JAMES G.
Published: April 2011

Application of 30 direct cotton dyes in aqueous salt solution to amyloid in human tissues yielded different staining patterns. Only dyes with good wool reserve without reactive metal atoms, i.e., dyes which stain cellulose but not proteins or polyamides, colored amyloid selectively without differentiation. Direct dyes for union fabrics, i.e., dyes which color cellulose and protein fibers, stained all tissue structures.

The effects of deamination and acetylation on the binding of direct dyes with good wool reserve indicate interaction of these dyes with hydroxyl groups of amyloid. The increase in intensity of staining upon addition of NaCl to the dye solution suggests a nonionic type linkage.

According to chemical data on the direct dyeing of cellulose, direct dyes are adsorbed by hydrogen bonding between hydroxyl groups of the polysaccharide chains and suitable groups in the dye. It seems probable that a similar mechanism is involved in the binding of these dyes by amyloid.

The selective staining of amyloid with direct dyes with good wool reserve, which do not color proteins, suggests that the polysaccharide moiety of amyloid may play a major role in the binding of direct dyes.
HISTOCHEMICAL STUDY OF CONTAMINANTS IN COMMERCIALLY AVAILABLE DIASTASES AND AMYLASES
Authors: PAEGLE ROLAND D.
Published: April 2011

No contaminants were detected with PAS, pyronin-Y and other staining methods in twice-crystallized alpha-amylase but most other commercially available diastases and amylases tested contained peptidase, ribonuclease, or both. Very small amounts of twice-crystallized alpha-amylase hydrolyzed glycogen in rat liver sections even at room temperature.
SEVERAL BIOCHEMICAL CRITERIA FOR EVALUATING β-GLUCURONIDASE LOCALIZATION
Authors: FISHMAN WILLIAM H. , GOLDMAN STEPHAN S. , GREEN SIDNEY
Published: April 2011

Biochemical evidence has been collected which supports the proposition that the appearance of the staining reaction in tissue sections incubated in Fishman-Baker substrate solution is dependent on the activity of β-glucuronidase. It was first shown by quantitative analysis that the glucuronic acid is liberated into the medium simultaneously with the deposition of ferric-hydroxyquinoline as the hydrolysis of the substrate, 8-hydroxyquinoline glucosiduronic acid, proceeded. This demonstration was made both in a gel-β-glucuronidase model system and in incubation mixtures containing tissue sections. Next, it was established that the staining reaction was dependent on pH in a manner indistinguishable from the pH dependence of purified rat liver β-glucuronidase acting on 8-hydroxyquinoline glucosiduronic acid. Finally, the staining reaction was invariably prevented by adding to the Fishman-Baker substrate solution an appropriate amount of a second glucosiduronic acid beside that of 8-hydroxyquinoline. This inhibition was produced by naphthol AS-BI-β-glucosiduronic acid, a histochemical substrate for β-glucuronidase. Next, it was observed that the addition of p-nitrophenyl β-glucosiduronic acid to the digest prepared for each staining reaction inhibited the staining reactions of both. The use of the mixed substrate test suggests itself for application to the critical study of other enzymes.
HISTOCHEMICAL RECOGNITION OF CALCIUM OXALATE
Authors: PIZZOLATO PHILIP
Published: April 2011

Of the workers applying the von Kóssa test to calcium oxalate crystals in animal tissue, about half reported blackening in silver nitrate on exposure to light. We found that "pure" calcium oxalate crystals, produced in rats by the administration of ammonium oxalate or ethylene glycol, gave a negative von Kóssa reaction. These crystals, however, were blackened when treated with a solution of silver nitrate in concentrated hydrogen peroxide. Other von Kóssa negative salts, such as CaF2 or CaSO4·2H2O, do not respond to the H2O2 and AgNO3 test.
THE STAINING OF TRIPHOSPHATASES BY THE CHELATE REMOVAL METHOD
Authors: BERG GEORGE G.
Published: April 2011

In the histochemical staining of enzymes by the chelate removal method, the substrate is a chelate, and the enzyme unmasks bound ions which are precipitated by suitable capturing ions in the incubating mixture. The new method allows the staining of phosphatases which act on substrates that are insoluble in the Gomori mixtures.

Three inorganic polyphosphatases were stained in the first application of the chelate-removal principle: neutral triphosphatase, alkaline triphosphatase, and alkaline pyrophosphatase. The incubating media had lead-polyphosphate chelates as substrates, with "tris" or barbital buffers. Orthophosphate was added to some solutions as an auxiliary capturing anion.

Alkaline triphosphatase as well as alkaline pyrophosphatase was demonstrated in the columnar epithelium of duodenal mucosa of mouse and rat, alkaline triphosphatase in the convoluted tubules of kidney, and neutral triphosphatase in bile canaliculi of liver. The properties of the three enzymes were examined, and the relation of the stains to biochemically defined enzymes of the inorganic polyphosphatase group was discussed.

The chelate removal method was also used for the histochemical demonstration of adenosine tri- and diphosphatase. Evidence for this mechanism of ATP-ase staining is presented as an addendum.