bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression of REG Protein During Cell Growth and Differentiation of Two Human Colon Carcinoma Cell Lines
Authors: Bernard-Perrone Françoise R. , Renaud Wanda P. , Guy-Crotte Odette M. , Bernard Patrice , Figarella Catherine G. , Okamoto Hiroshi , Balas Daniel C. , Senegas-Balas Françoise O.
Published: April 2011

We localized REG protein in Paneth cells and nonmature columnar cells of the human small intestinal crypts and speculated that this protein was associated with growth and/or differentiation. The aim of this study was to determine whether REG protein is present in two human colon cancer cell lines that exhibit enterocytic differentiation after confluence and to investigate changes in the level of its expression during growth and differentiation. Results were compared to those obtained on cells that remain undifferentiated. Western immunoblotting and immunofluorescence demonstrated the presence of REG protein in the three cell lines. With the antisera against human REG protein, the staining was diffusely spread throughout the cytoplasm at Day 2, and after Days 3-4 it appeared to have migrated to cell boundaries. After confluence, we observed only a punctate staining array along cell boundaries, which disappeared at Day 15. REG mRNA expression was demonstrated by RTPCR and REG mRNA hybridization until Day 13, but not after, in the three cell types. REG protein may be involved in cellular junctions. Its presence appears to be associated with the cell growth period and the protein must be downregulated when growth is achieved and differentiation is induced.
Presence of H Type 3/4 Chains of ABO Histo-blood Group System in Serous Cells of Human Submandibular Gland and Regulation of Their Expression by the Secretor Gene (FUT2)
Authors: Liu Yuhua , Fujitani Noboru , Koda Yoshiro , Soejima Mikiko , Kimura Hiroshi
Published: April 2011

We have investigated by immunochemistry the distribution of H Type 3/4 chains of the ABO histo-blood group system in human submandibular gland using a monoclonal anti-H MBr1 antibody specific for H Type 3/4 chains, and have found the expression of H Type 3/4 chains was mainly in the serous cells. Serous cells from secretors were stained by MBr1 but not by anti-A and anti-B antibodies, whereas serous cells from nonsecretors exhibited a negative reaction with MBr1. Mucous cells were not stained by MBr1. Only a few striated duct cells showed a weak reaction with anti-H MBr1. These results suggested that the H Type 3/4 chains were distributed predominantly in the serous cells of the human submandibular gland and that secretor Type α (1,2) fucosyltransferase (Se enzyme) controlled the synthesis of H Type 3/4 chains in vivo. Saliva also contained H Type 3/4 chains, which were controlled by the secretor gene ( FUT2). The differences in the distributions of H Type 1, H Type 2, and H Type 3/4 chains of the ABO histo blood group system in the submandibular gland are discussed.
Immunocytochemical Detection of Ornithine Decarboxylase
Authors: Schipper Raymond G. , Romain Nadine , Otten Adrianus A. , Tan Jing , Lange Will P. , Verhofstad Albert A. J.
Published: April 2011

Ornithine decarboxylase (ODC), a regulatory enzyme of polyamine biosynthesis, is involved in cell growth and differentiation. Lack of information about the exact cellular and subcellular localization of ODC is one of the main obstacles to precise interpretation of the biological roles of the ODC/polyamine system. Here we describe the development and optimization of an immunocytochemical method to detect ODC in cells and tissues. For this purpose a monoclonal antibody (MP16-2) against a defined epitope of ODC protein was developed. Specificity of the antibody for ODC was substantiated by Western blotting and ELISA analysis using cell and tissue homogenates. In cultured cells, optimal staining results were obtained after fixation with crosslinking fixatives followed by permeabilization with methanol. In rat tissues, ODC immunoreactivity was best preserved in paraffin sections fixed with Bouin's fixative. Antigen retrieval using SDS and citrate buffer substantially increased ODC immunostaining and decreased background staining. Localization studies of ODC in different cell lines showed that strongest staining for ODC was found in the nucleoplasm of mitotic cells, whereas confluent cells showed moderate perinuclear staining. Immunocytochemical studies of various rat tissues showed high cytoplasmic immunostaining of ODC in epithelial cells of kidney, prostate, and adrenal medulla of testosterone-treated rats, in glandular epithelium of small intestine, and in pancreas of neonatal and adult rats.
Loss of Heterozygosity of Tumor Suppressor Genes (p16, Rb, E‐cadherin, p53) in Hypopharynx Squamous Cell Carcinoma
Authors: Lee Sang‐Hyuk , Lee Nam‐Hoon , Jin Sung‐Min , Rho Young‐Soo , Jo Sung‐Jin
Published: April 2011

Objective

Microsatellite alterations, especially those that cause loss of heterozygosity (LOH), have recently been postulated as a novel mechanism of carcinogenesis and a useful prognostic factor in many kinds of malignant tumors. However, few studies have focused on a specific site, hypopharynx. The aim of this study was to evaluate the relationship between LOH and hypopharyngeal squamous cell carcinoma (HPSCC).

Study Design

Laboratory‐based study.

Setting

Integrated health care system.

Subjects and Methods

Matched normal and cancerous tissues from 30 patients with HPSCC were examined for LOH in 4 tumor suppressor genes (TSGs) (p16, Rb, E‐cadherin, and p53) at loci 9p21, 13q21, 6q22, and 17p13, respectively, using microsatellite markers amplified by polymerase chain reaction. The results for each loci were compared with clinicopathological features.

Results

Among the 30 cases, 26 (86.7%) exhibited LOH, with the most common alteration being LOH at p53 (52.6%). Significantly higher rates of LOH detection were seen in Rb, p53, and the LOH‐high group (cases where 2 or more loci with LOH were found) in cases of lymph node metastasis. Compared with stage I and II carcinoma, tumors of stages III and IV had significantly higher frequencies of LOH in Rb, p53, and the LOH‐high group. However, the presence of LOH was not significantly correlated with survival.

Conclusion

These results suggest that LOH in TSGs such as Rb and p53 may contribute to the development and progression of HPSCC. The presence of LOH in the primary tumor may also be predictive of lymph node metastasis.
Third Windows as a Cause of Failure in Hearing Gain after Exploratory Tympanotomy
Authors: Kim Sang Cheol , Lee Won‐Sang , Kim Minbum , Jeon Ju Hyun , Kim Jinna , Choi Jae Young
Published: April 2011

Objective

Exploratory tympanotomy (ET) can be performed selectively on patients who have air‐bone gaps (ABGs) without any apparent external or middle ear abnormalities for diagnosis and treatment of their conductive hearing loss. However, surgeons cannot always find middle ear problems. It has recently been reported that pathologic third windows in inner ears can cause ABGs. Therefore, the authors conducted this study to evaluate the inner ear anatomical problems of patients who had undergone ET.

Study Design

Case series with chart review.

Setting

Tertiary center.

Subjects and Methods

The authors included 51 consecutive patients with ABGs ≥20 dB who had undergone ET at Yonsei University Severance Hospital between 2005 and 2009. In the preoperative evaluations, all 51 patients showed normal drums on otoscopy, and no abnormal middle ear problems that could cause ABGs were detected on temporal bone computed tomography (TBCT). The authors retrospectively reevaluated the hearing results and the causes of their conductive hearing loss, including the inner ear problems, through a review of their TBCT.

Results

Six of the 51 patients were found to have inner ear abnormalities with possible pathologic third windows. Four had enlarged vestibular aqueducts (EVAs), and 2 had superior semicircular canal dehiscence (SSCD). All patients with pathologic third windows failed to improve their hearing.

Conclusion

About 12% of the patients had inner ear anomalies, which could possibly cause ABGs. Inner ear conductive components might lead to frustrating results. Therefore, surgeons must take inner ear conductive components into account before attempting ET in patients with inner ear anomalies.
Dynamic Interactions between Orexin and Dynorphin May Delay Onset of Functional Orexin Effects: A Modeling Study
Authors: Williams Katherine S. , Diniz Behn Cecilia G.
Published: March 2011

Orexin (also known as hypocretin) neurons play a key role in regulating sleep-wake behavior, but the links between orexin neuron electrophysiology and function have not been explored. Orexin neurons are wake-active, and spiking activity in orexin neurons may anticipate transitions to wakefulness by several seconds. However, it is suggested that while the orexin system is necessary to maintain sustained wake bouts, orexin has little effect on brief wake bouts. In vitro experiments investigating the actions of orexin and dynorphin, a colocalized neuropeptide, on orexin neurons indicate that the dynamics of desensitization to dynorphin may represent a mechanism for modulating local network activity and resolving the apparent discrepancy between the onset of firing in orexin neurons and the onset of functional orexin effects. To investigate the role of dynorphin on orexin neuron activity, a Hodgkin-Huxley—type model orexin neuron was developed in which baseline electrophysiology, orexin/dynorphin action, and dynorphin desensitization were closely tied to experimental data. In this model framework, model orexin neuron responses to orexin/dynorphin action were calibrated by simulating the physiologic effects of static orexin and dynorphin bath application on orexin neurons. Then behavior in a small network of model orexin neurons was simulated with pure orexin, pure dynorphin, or combined orexin and dynorphin coupling based on the mechanisms established in the static case. It was found that the dynamics of desensitization to dynorphin can mediate a clear shift from a network in which firing is suppressed by dynorphin-mediated inhibition to a network of neurons with high firing rates sustained by orexin-mediated excitation. The findings suggest that dynamic interactions between orexin and dynorphin at transitions from sleep to wake may delay onset of functional orexin effects.
Elimination of Short-Day Melatonin Signaling Accelerates Gonadal Recrudescence but Does Not Break Refractoriness in Male Turkish Hamsters
Authors: Jarjisian Stephan G. , Zucker Irving
Published: March 2011

Long days stimulate and short days (SDs) inhibit the reproductive axis of photoperiodic rodents. In long-day Turkish hamsters, unlike most other rodents, elimination of pineal melatonin secretion by constant light or pinealectomy initiates a cycle of gonadal involution and recrudescence outwardly similar to that induced by short days. The present study assessed whether short days and constant light induce the seasonal reproductive cycle via common or different interval timing mechanisms. Male hamsters that had undergone gonadal involution in SDs for 8 or 14 weeks were treated with LL for 14 and 8 weeks, respectively. If SDs and LL act via independent mechanisms, then gonadal quiescence of SD-regressed males, which normally lasts 10 weeks, might be extended by LL treatment; alternatively, if SDs and LL act on the same timer, or the timer cannot be retriggered, then LL will not extend the duration of reproductive quiescence. Neither of these outcomes materialized. Instead, male hamsters exposed to LL while reproductively quiescent exhibited accelerated gonadal recrudescence. Extended LL treatment did not restore responsiveness to SDs in photorefractory hamsters. In Turkish hamsters, photoperiodic history determines whether constant light inhibits or stimulates the hypothalamic-pituitary-testicular axis.
Purinergic Signals Regulate Daily S-Phase Cell Activity in the Ciliary Marginal Zone of the Zebrafish Retina
Authors: Jimena Ricatti Maria , Battista Ariadna Gabriela , Zorrilla Zubilete Maria , Faillace Maria Paula
Published: March 2011

Regeneration and growth that occur in the adult teleost retina have been helpful in identifying molecular and cellular mechanisms underlying cell proliferation and differentiation. Here, it is reported that S-phase cell number, in the ciliary marginal zone (CMZ) of the adult zebrafish retina, exhibits day-night variations with a mid-light phase peak. Oscillations persist for 24 h in constant darkness (DD), suggesting control by a circadian component. However, variations in the S-phase nuclei number were rapidly dampened and not present during and after a second day in DD. An ADPβS treatment significantly enhanced S-phase activity at night to mid-light levels, as assessed by in vivo BrdU incorporation in a 2-h interval. Moreover, daylight increase in S-phase cell number was completely abolished when extracellular nucleotide levels or their extracellular hydrolysis by ectonucleoside triphosphate diphosphohydrolases (NTPDases) were significantly disrupted or when a selective antagonist of purinergic P2Y1 receptors was intraocularly injected before BrdU exposure. Extracellular nucleotides and NTPDase action were also important for maintaining nocturnal low levels of S-phase activity in the CMZ. Finally, we showed that mRNAs of NTPDases 1, 2 (3 isoforms), and 3 as well as of P2Y1 receptor are present in the neural retina of zebrafish. NTPDase mRNA expression exhibited a 2-fold increment in light versus dark conditions as assessed by quantitative RT-PCR, whereas P2Y1 receptor mRNA levels did not show significant day-night variations. This study demonstrates a key role for nucleotides, principally ADP as a paracrine signal, as well as for NTPDases, the plasma membrane—bound enzymes that control extracellular nucleotide concentration, for inducing S-phase cell entry in the CMZ—normally associated with retinal growth—throughout the light-dark cycle.
Lab Mice in the Field: Unorthodox Daily Activity and Effects of a Dysfunctional Circadian Clock Allele
Authors: Daan Serge , Spoelstra Kamiel , Albrecht Urs , Schmutz Isabelle , Daan Moritz , Daan Berte , Rienks Froukje , Poletaeva Inga , Dell'Omo Giacomo , Vyssotski Alexei , Lipp Hans-Peter
Published: March 2011

Daily patterns of animal behavior are potentially of vast functional importance. Fitness benefits have been identified in nature by the association between individual timing and survival or by the fate of individuals after experimental deletion of their circadian pacemaker. The recent advances in unraveling the molecular basis of circadian timing enable new approaches to natural selection on timing. The investigators report on the effect and fate of the mutant Per2Brdm1 allele in 4 replicate populations of house mice in a seminatural outside environment over 2 years. This allele is known to compromise circadian organization and entrainment and to cause multiple physiological disturbances. Mice ( N = 250) bred from Per2Brdm1 heterozygotes were implanted subcutaneously with transponders and released in approximately Mendelian ratios in four 400 m2 pens. An electronic system stored the times of all visits to feeders of each individual. The study first demonstrates that mice are not explicitly nocturnal in this natural environment. Feeding activity was predominantly and sometimes exclusively diurnal and spread nearly equally over day and night under the protective snow cover in winter. The effect of Per2Brdm1 on activity timing is negligible compared to seasonal changes in all genotypes. Second, the Per2Brdm1 allele did not have persistent negative effects on fitness. In the first year, the allele gradually became less frequent by reducing survival. New cohorts captured had the same Per2Brdm1 frequency as the survivors from previous cohorts, consistent with an absence of an effect on reproduction. In the second year, the allele recovered to about its initial frequency (0.54). These changes in selective advantage were primarily due to female mice, as females lived longer and the sex ratio dropped to about 25% males in the population. While it is unknown which selective advantage led to the recovery, the results caution against inferences from laboratory experiments on fitness consequences in the natural environment. It also demonstrates that the activity of mice, while strictly nocturnal in the laboratory, may be partially or completely diurnal in the field. The new method allows assessment of natural selection on specific alleles on a day-today basis.
Updating the Outcome
Authors: Anderson Eric
Published: March 2011

In this article I report findings from interviews with 26 openly gay male athletes who came out between 2008 and 2010. I compare their experiences to those of 26 gay male athletes who came out between 2000 and 2002. The athletes in the 2010 cohort have had better experiences after coming out than those in the earlier cohort, experiencing less heterosexism and maintaining better support among their teammates. I place these results in the context of inclusive masculinity theory, suggesting that local cultures of decreased homophobia created more positive experiences for the 2010 group.
The Notion of Quality in DPEP Pedagogical Interventions
Authors: Dhankar Rohit
Published: March 2011

The District Primary Education Programme (DPEP) has worked in 18 states in India with the agenda to enhance the quality of classroom interaction and the teaching-learning materials used. This paper examines how ‘quality’ is understood within DPEP, and the educational principles that guide DPEP efforts and training methods, based on reports of training workshops, examining textbooks and materials and first-hand field experience. A proposal is made for an alternative way of looking at ‘quality’ and efforts to realise it.
A CYTOLOGICAL INVESTIGATION OF THE MITOCHONDRIA OF THREE STRAINS OF SALMONELLA TYPHOSA
Authors: DAVIS JOHN C. , WINTERSCHEID LOREN C. , HARTMAN PHILIP E. , MUDD STUART
Published: April 2011

Various supravital mitochondrial indicators have been used on three strains of S. typhosa. Some variation in the pattern of staining was noted, which demonstrates the existence of a gradient of mitochondrial reactivities. The mitochondria were differentiated from nuclei and cell wall, and Showed no structural relationship to terminal flagellar spheres.
A COMPARISON OF THE STAINING OF LIPID DROPLETS IN THE MOUSE OVARY BY THE SCHIFF REACTION AND BY THE ASHBEL-SELIGMAN CARBONYL REACTION
Authors: DEANE HELEN WENDLER , ANDREWS JEAN S.
Published: April 2011

Fresh-frozen sections of mouse ovary, briefly fixed in formalin, fail to stain with fuchsin-sulfurous acid (the Schiff reagent) or the Ashbel-Seligman carbonyl reagents. When the sections are treated for 10 minutes with mercuric chloride and then stained, the cytoplasm of the cells acquires a faint coloration—the plasmal reaction. When the sections are washed in water or dilute alcohol for 3 or 24 hours, however, lipid droplets become increasingly stainable. The results of treatment of the sections with halogens, oxidants and reductants suggest that the aldehydic substances which react with the Schiff reagent are all derived from unsaturated lipids but that, in addition, ketonic substances are formed or unmasked which are reactive to the hydrazide alone. Whether or not the ketonic lipids are steroidal has not been demonstrated.
THE ACTION OF FORMALDEHYDE ON RAT BRAIN LIPIDS
Authors: DEIERKAUF F. A. , HESLINGA F. J. M.
Published: April 2011

The influence of formaldehyde on rat brain lipid was investigated by comparing the analysis of lipids from fresh rat brain with the analysis from rat brains fixed for 110 days in a formaldehyde solution.

A quantitative analysis of the lipids using column and paper chromatography is described.

Sphingomyclin, cerebrosides and sulphatides were not attacked. Only a slight diminution of phosphatidylserine was indicated. Phosphoinositides remain fairly constant. Of lecithin about 25% was converted to lyso-lecithin. The phosphatidylethanolamine and lyso-phosphatidylethanolamine fractions suffered a marked diminution and the reaction with formaldehyde was noticeable in a change of chromatographic behaviour. The fraction containing cholesterol, cholesterol esters, glycerides, and fatty acids increased, the increase being correlated with the decrease of other fractions by the formation of lyso compounds.

After storing tissues for months in formaldehyde solutions a reliable analysis of the lipid content is still possible.
THE REACTION OF OSMIUM TETROXIDE WITH LECITHIN: APPLICATION OF STAINING PROCEDURES
Authors: RIEMERSMA JELLE C. , BOOIJ HEIN L.
Published: April 2011

To obtain additional insight into the chemical reactions occuring furing fixation of tissue lipids by osmium tetroxide, the reaction between osmium tetroxide and lecithin has been studied. Procedures have been developed whereby quantitative information may be obtained about the nature of this reaction.

One series of experiments was designed to arrive at the stoichiometry of the reaction and its time-course. To this end air-dried lecithin spots on cover glasses or on filter paper were exposed to oxmium tetroxide vapor. The weight increase of the lecithin spots was determined, and related to time of exposure to osmium tetroxide and to the quantity of lecithin present. Under the conditions chosen the binding of osmium tetroxide rapidly reaches a limiting value; thus the reaction appears to proceed to a definite end-point. The number of double bonds in the lecithin molecule, as determined by gas chromatography, corresponds with the number of moles of osmium tetroxide bound per mole of lecithin. This supplements earlier spectrophotometric findings, and confirms part of the reaction scheme proposed by Criegee for the interaction between osmium tetroxide and unsaturated compounds generally.

The chemical properties of lecithin, and of other polar lipids, are determined not only by the presence of unsaturated linkages but also by the presence of an ampho-ionic group. Earlier research has demonstrated that this group is responsible for the staining properties of lecithin with anionic dyes, for instance, Brilliant Scarlet 3 R (Ponceau 4R or 4RS, Colour Index No. 16255). In this paper evidence is presented to show that the stainability of lecithin spots by Brilliant Searlet 3 R, under specified conditions, decreases markedly with increasing time of exposure to osmium tetroxide. It appears likely that as a result of the reaction with osmium tetroxide the lecithin polar group has changed properties.

After studying the staining properties of lecithin towards an anionic dye, in relation to time of exposure to osmium tetroxide, its staining capacity for a representative cationic dye was similarly studied. Evidence is presented demonstrating an increased binding capacity of exposed lecithin spots to Brilliant Green (Colour Index no 42040). After considering a few possible interpretations we suggest that the simplest hypothesis to account for this phenomenon is a partial removal of the positive charge of lecithin, whereby the negatively charged groups are freed for interaction with positive dye molecules.
SYMPOSIUM: SYNTHETIC PROCESSES IN THE CELL NUCLEUS. V. GLYCOGEN ACCUMULATION IN THE NUCLEUS
Authors: HIMES MARION M. , POLLISTER ARTHUR W.
Published: April 2011

Hepatic intranuclear glycogen of the larva of Rana pipiens has been studied by the periodic acid Schiff reaction (controlled by diastase digestion), by electron microscopy, and by interference and phase microscopy. The glycogen occurs sporadically as one or two masses in the central part of the nucleus. The absence of nuclear envelope and of typical non-glycogen cytoplasmic elements shows that the nuclear glycogen inclusions are not cytoplasmic areas that have been enclosed within the nucleus. The central position, difference in rate of resorption, and other evidence indicates that the nuclear glycogen has been synthesized in situ. Comparison with the nucleolus suggests that the nuclear glycogen body is formed as a result of activity at a specific chromosomal site.
How the Brain Shapes Deception
Authors: Abe Nobuhito
Published: April 2011

How do people tell a lie? One useful approach to addressing this question is to elucidate the neural substrates for deception. Recent conceptual and technical advances in functional neuroimaging have enabled exploration of the psychology of deception more precisely in terms of the specific neuroanatomical mechanisms involved. A growing body of evidence suggests that the prefrontal cortex plays a key role in deception, and some researchers have recently emphasized the importance of other brain regions, such as those responsible for emotion and reward. However, it is still unclear how these regions play a role in making effective decisions to tell a lie. To provide a framework for considering this issue, the present article reviews current accomplishments in the study of the neural basis of deception. First, evolutionary and developmental perspectives are provided to better understand how and when people can make use of deception. The ensuing section introduces several findings on pathological lying and its neural correlate. Next, recent findings in the cognitive neuroscience of deception based on functional neuroimaging and loss-of-function studies are summarized, and possible neural mechanisms underlying deception are proposed. Finally, the priority areas of future neuroscience research—human honesty and dishonesty—are discussed.
Multisectorial Afghan Perspectives on Girl Child Marriage
Authors: Raj Anita , Gomez Charlemagne S. , Silverman Jay G.
Published: March 2011

The objective of this study was to identify Afghan perspectives on the causes of and potential solutions to child and forced marriage in the country. Open-ended interviews ( N = 102 interviews) were conducted with religious leaders, police, teachers, and nongovernmental organizations (NGO) and government officials in Kabul, Jalalabad, and Mazar. Informants reported recognition of the poor social and health consequences of these practices for mothers and infants, citing poverty, tradition, conflict-related insecurity, low status of women, and ignorance of religious and civil laws as causes of these practices. Recommended solutions centered on child marriage prevention; most informants felt little can be done for married girls.
OXIDATIVE AND HYDROLYTIC ENZYMES OF HOMOZYGOUS DYSTROPHIC AND HETEROZYGOUS MUSCLE OF THE HOUSE MOUSE
Authors: FENNELL RICHARD A. , WEST WILLIAM T.
Published: April 2011

Histochemical procedures were used for the identification of oxidative and hydrolytic enzymes of homozygous dystrophic (genotype, dydy) and heterozygous (genotype, Dydy) muscles of the house mouse.

In adult heterozygous muscle and muscle of a Swiss albino mouse there was usually a reciprocal relationship between phosphorylase and lactate dehydrogenase, i.e., weak lactate dehydrogenase reactions in the majority of fibers of the vastus lateralis and a stronger phosphorylase reaction. Weak to strong lactate dehydrogenase reactions were identified in fibers of the rectus femoris. In heterozygous muscle of young mice 20 days of age, a reciprocal relationship between lactate dehydrogenase and phosphorylase was not observed.

Reciprocity between phosphorylase and lactate dehydrogenase was not always evident in fibers of homozygous muscles, e.g., groups of homozygous fibers frequently exhibited both strong phosphorylase and lactate dehydrogenase reactions whereas other fibers reacted weakly for phosphorylase or not at all and strongly for lactate dehydrogenase. Small fibers reacted either weakly or strongly for dehydrogenases.

One of the striking features which homozygous fibers usually showed was uniformly strong dehydrogenase reactions (e.g. lactate, glutamate, succinate) and diphosphopyridine nucleotide linked diaphorase reactions. Strong glucose-6-phosphate dehydrogenase reactions were frequently identified in homozygous dystrophic fibers whereas weak reactions or no reactions at all were seems in heterozygous fibers.

Atrophic fibers of homozygous dystrophic muscles exhibited intense indoxyl acetate esterase and acid phosphatase reactions. Weak adenosine triphosphatase reactions were seen in fibers of both heterozygous and homozygous dystrophic muscle, and strong reactions in connective tissues and blood vessels.

Interfibrillar connective tissue of homozygous muscle was abundant and exhibited intense acid phosphatase reactions, whereas connective tissues of heterozygous muscle were less concentrated and exhibited weak acid phosphatase reactions.
ANALYTICAL CYTOLOGY OF MUCIN PRODUCTION: MUCINOUS CYTOPLASMIC GRANULES IN A MYXOMATOUS TUMOR OF HAMSTER
Authors: ROSAN ROBERT C. , SPRING ELINOR J.
Published: April 2011

Cell counts and differential stains indicate that about 90% of cells in viable areas of a homologous myxomatous tumor of hamster check-pouch are probably of one single type. The cytoplasm of these cells contains granules which stain with periodic acid Schiff reaction and are different from granules of mast cells in the same sections. Most of the parenchymal ground substance gives the alcian blue reaction for acid mucopolysaccharides, but some stains with the periodic acid Schiff reaction in the same manner as the cytoplasmic granules.
FURTHER OBSERVATIONS ON HISTOCHEMICAL ESTERASE AND AMIDASE ACTIVITIES WITH SIMILARITIES TO TRYPSIN
Authors: HOPSU VÄINÖ K. , GLENNER GEORGE G.
Published: April 2011

When the histochemical substrates benzoyl-dl-arginine β-naphthylamide, benzoyl-dl-lysine β-naphthylamide, Naphthol AS and AS-D ε-aminocaproate, and Naphthol AS β-alanyl glycinate were tested with tissue sections and as substrates for purified enzymes, a species-limited enzyme activity having substrate specificity and enzymic kinetic and affector characteristics strikingly similar to those of bovine trypsin was noted in human mast cells. The ester substrates were hydrolyzed in several other tissue sites, and, in those tested, hydrolysis appeared to be related to the activity of an aliesterase(s) with substrate specificity and affector characteristics markedly different from those of bovine trypsin. Although Naphthol AS chloroacetate has been defined as a substrate for a chymotrypsin-like enzyme in rat mast cells, the definite possibility exists that its hydrolysis in human mast cells in fresh frozen tissue sections may be due at least its part to the activity of a trypsin-like enzyme.
THE ROLE OF IRON IN THE MECHANISM OF EXPERIMENTAL CALCIFICATION
Authors: GABBIANI G. , TUCHWEBER B.
Published: April 2011

Calcification of different organs in the rat, induced by challenge with various iron compounds after sensitization with dihydrotachysterol, was studied histochemically. Iron is demonstrable in the target organs before calcification but apparently, in the described experimental conditions, it can attract calcium only when it is extracellular.

The possible relationship between ferrification and calcium deposition is briefly discussed.
ELECTRON MICROSCOPIC CYTOCHEMISTRY OF MELANOSOMES AND MITOCHONDRIA
Authors: MISHIMA YUTAKA
Published: April 2011

The electron histochemical properties characteristic of melanosomes as distinguished from mitochondria are presented. Our modified ammoniated silver nitrate reaction for electron microscopy reveals an essential chemical property of melanosomes which is also possessed by melanin granules, but is not possessed by any mitochondria. This property is the reduction of ammoniated silver nitrate to the metallic state.

Furthermore, melanosomes, and not mitochondria, are the only organelles which acquire the appearance of typically mature melanin granules after the electron microscopic dopa reaction.
DEMONSTRATION OF PEROXIDASE ACTIVITY IN TISSUE SECTIONS
Authors: WACHSTEIN MAX , MEISEL ELIZABETH
Published: April 2011

By using an improved benzidine technique, peroxidase activity can be demonstrated in various locations in mammalian tissues. A relatively formalin resistant enzyme is found in hemoglobin and is also associated with mitochondria of striated muscle and heart. A somewhat less formalin resistant peroxidase occurs in the granules of myeloid and mast cells. A relatively formalin sensitive peroxidase is present in a number of additional locations, e.g. the acinar cells in thyroid and salivary gland, the medulla of the kidney, in hair follicles of the guinea pig skin and Kupffer cells of the liver.
A METHOD FOR THE CYTOCHEMICAL DEMONSTRATION OF SODIUM-ACTIVATED ADENOSINE TRIPHOSPHATASE
Authors: MCCLURKIN lOLA T.
Published: April 2011

(1) In the histochemical method for adenosine triphosphatase, when magnesium and ATP are in equivalent concentrations, and sodium is present in the medium, staining occurs in the tissues. When sodium is omitted from this medium some structures—previously stained in the presence of sodium—fail to stain.

(2) Ouabain inhibits staining when magnesium and ATP are equivalent and sodium is present in the medium.

(3) The effects of calcium, potassium and sodium on staining with the proposed medium parallel the effects of these ions on activity of sodium-activated ATPase in vitro.

(4) When magnesium ions and ATP are not equivalent in the substrate, sodium and ouabain fail to affect staining.

(5) Certain tissue structures—such as the axonic nerve ending—which have been shown to contain sodium-activated ATPase in vitro, stain in the presence of sodium when magnesium and ATP are equivalent, but fail to stain when sodium is omitted from this medium.
THE ENZYMATIC HYDROLYSIS OF AMINO ACID β-NAPHTHYLAMIDES: I. PREPARATION OF AMINO ACID AND DIPEPTIDE β-NAPHTHYLAMIDES
Authors: GLENNER G. G. , COHEN L. A. , FOLK J. E.
Published: April 2011

The acid salts of amino acid β-naphthylamides have been found to have greater stability and solubility than their corresponding free bases. For the purpose of applying these substrates in subsequent studies in histochemical and biochemical systems, the syntheses and characteristics of a large variety of previously unreported intermediates, acid salts and free bases of amino acid β-naphthylamides and their analogues are described.
INDOPHENOL OXIDASE AND PEROXIDASE ACTIVITIES IN MAMMALIAN ERYTHROCYTES
Authors: SIEGEL ISRAEL
Published: April 2011

Cytochrome oxidase and peroxidase activities in mammalian red cells are suggested by G- and M-Nadi reactions in rabbit, mouse, and human erythrocytes. Peroxidase activity was present in both the hemoglobin and the nonhemoglobin constituents of the red cells. Removal of oxygen stopped the G-Nadi reaction but not the M-Nadi reaction. Potassium cyanide and sodium azide at a concentration of 10–3-10–4 M partially inhibited G- and M-Nadi reactions. Microscopic examination of reacting cells revealed accumulation of reaction products at the cell periphery. This was not evident when the cells were incubated in a suspension of indophenol blue.

Mammalian erythrocytes reduced the tetrazolium salt nitro-BT. Reduction was augmented by removal of oxygen and by cytochrome oxidase inhibitors. Accumulation of formazan at the surface of reacting cells was evident upon microscopic examination.

The Nadi reactions were similar whether Ehrlich ascites tumor cells or stroma of hemolyzed erythrocytes were used. Reactions of stroma or whole erythrocytes with nitro-BT were essentially similar. Fixation or heating slowed or abolished all the reactions described.
LOCALIZATION OF PROLACTIN IN RAT PITUITARY AND IN A TRANSPLANTABLE MAMMOTROPIC PITUITARY TUMOR USING FLUORESCENT ANTIBODY
Authors: EMMART E. W. , BATES R. W. , TURNER W. A.
Published: April 2011

By means of fluorescent antiprolactin globulin and with appropriate controls, lactogenic hormone has been localized in granules and mitochondnia of acidophils of normal rat pituitary and in a transplantable rat pituitary tumor. Cells containing prolactin have been identified as acidophils by their reaction with orange C and by positive staining of their cytoplasmic granules with acid fuchsin in the Russell technique. Immunochemical data is presented reaffirming the heterogenous character of purified ovine prolactin. Evidence that this is associated with polymerization of prolactin is discussed.
COMPARATIVE STUDY OF MAMMALIAN ESTERASES BY THE ZYMOGRAM METHOD
Authors: COUTINHO HÉLIO B. , PADILHA MANOEL C. S. , GOMES JURANDY M. , ALVES J. J. ALMEIDA
Published: April 2011

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.
VARIATIONS IN DEHYDROGENASE AND NEOTETRAZOLIUM REDUCTASE (DIAPHORASE) ACTIVITIES IN VARIOUS BUFFERS STUDIED QUANTITATIVELY IN FROZEN SECTIONS
Authors: MCCABE MARY , MAPLE A. J. , JONES G. R. N.
Published: April 2011

A quantitative study has been made in frozen sections of rat liver of the behavior of certain NAD+- and NADP+-linked dehydrogenases and the corresponding neotetrazolium reductases under histochemical conditions in phosphate, Tris and metaborate buffers. The rates of reaction were considerably decreased, though to variable extents, its metaborate buffer, with the exception of NADH-neotetrazolium reductase and the NADP+-linked glucose 6-phosphate and 6-phosphogluconate dehydrogenases. Glyceraldehyde 3-phosphate dehydrogenase could not be demonstrated with neotetrazolium as electron acceptor. Formazan production by both lactate and β-hydroxybutyrate dehydrogenases in phosphate and Tris buffers was almost as rapid as by NADH-neotetrazolium reductase, suggesting that in parts of the section the latter reaction was the rate-limiting step in these two cases.
Peer interaction patterns among adolescents with autistic spectrum disorders (ASDs) in mainstream school settings
Authors: Humphrey Neil , Symes Wendy
Published: April 2011

The aim of the current study was to document the peer interaction patterns of students with autistic spectrum disorders in mainstream settings. Structured observations of a group of 38 adolescents with ASD drawn from 12 mainstream secondary schools were conducted over a two-day period and data compared with those of school, age, and gender matched comparison groups of 35 adolescents with dyslexia and 38 with no identified special educational needs (the ASD and dyslexia groups were also matched on SEN provision). Frequency and duration of peer interaction behaviours were coded. In terms of duration, multivariate analyses of variance (MANOVAs) indicated that participants with ASD spent more time engaged in solitary behaviours, less time engaged in co-operative interaction with peers, and more time engaging in reactive aggression towards peers than either comparison group. In terms of frequency, similar patterns emerged, but additionally participants with ASD engaged in fewer instances of rough/vigorous play, and were subject to more instances of social initiation and instrumental verbal aggression by peers than either comparison group. The findings of the current study support the authors’ theoretical model of peer group interaction processes for individuals with ASD, and have implications for both social skills training and the development of peer awareness and sensitivity. Limitations are noted.
A NEW ION ASSOCIATION TECHNIQUE FOR DEMONSTRATING POLYANIONS IN TISSUE SECTIONS
Authors: ZUGIBE FREDERICK T. , FINK MARILYN L.
Published: April 2011

A new ion association technique was developed in our laboratory to demonstrate polyanions histochemically. The identification of the polyanions was effected in two steps: (1) the formation of a polyanion-quaternary ammonium complex following treatment with a quaternary salt and (2) subsequent formation of a red ion association complex with the quaternary moiety following treatment with ferric thiocyanate solution. Chemical studies indicated that the ion association complexes are cetyltrimethylammonium-hexathiocyanatoferrate (iii) and cetylpyridinium-hexathiocyanatoferrate (iii). These are red in color, very soluble in organic solvents such as chloroform, ethanol, methanol, acetone and xylene and insoluble in water.
LOCALIZATION BY IMMUNOFLUORESCENCE OF AMYLASE, TRYPSINOGEN AND CHYMOTRYPSINOGEN IN THE ACINAR CELLS OF THE PIG PANCREAS
Authors: YASUDA KENJIRO , COONS ALBERT H.
Published: April 2011

The localization of α-amylase in the pig pancreas, and of chymotrypsinogen and trypsinogen in the beef pancreas, has been investigated by immunofluorescence, using antisera prepared in rabbits against commercial crystalline enzyme preparations. Porcine α-amylase was abundant in almost every acinar cell, being distributed as fine granular material in the basal part of the cell, and also in some of the zymogen granules in the juxta-nuclear region. It was visible also in the Golgi zone. Bovine chymotrypsinogen and trypsinogen were present in the presumably less mature zymogen granules in the cytoplasm around the nucleus, but the granules near the lumen failed to react with the antisera, perhaps because they were hidden by superficial material. The specificity of the anti-amylase serum was high, that of the anti-chymotrypsinogen and anti-trypsinogen lower because of cross reactions with each other. In both porcine and bovine pancreas, the acinar cells were virtually all similar, indicating functional similarity to the extent of these observations.
STUDIES IN HISTOCHEMISTRY. LXXXV HISTOCHEMICAL DEMONSTRATION OF PROTEIN WITH BROMSULFALEIN
Authors: SILVERMAN LLOYD , GLICK DAVID
Published: April 2011

Bromsulfalein has been shown to provide an intense blue stain for protein with some advantages over other acid dyes that have been used similarly. The procedure is simple and requires no unusual materials. The previously established stoichiometric nature of the reaction between the anionic dye and the cationic groups of protein offer the potential use of the stain for measurement.
THE KINETICS OF CHOLINESTERASES MEASURED FLUOROMETRICALLY
Authors: SIEGEL GEORGE J. , LEHRER GERARD M. , SILIDES DEMETRA
Published: April 2011

A new, simple, sensitive method is described for the fluorometric assay of cholinesterase activity based upon the hydrolysis of 1-naphthyl esters and the measurement of the fluorescence of 1-naphthol. This allows the study of the kinetics of cholinesterases and inhibitors with histochemical substrates and permits assessment of the parameters of the enzyme reaction under conditions approximating those in the histochemical system. 1-Naphthylacetate is a substrate for AChE3 and ChE, while 1-naphthyl butyrate is selective for ChE. The application of the procedure to the study of inhibition by hydrolyzable as well as nonhydrolyzable, nonfluorogenic, inhibitors is demonstrated. ACh was found to be a mixed inhibitor of eel AChE in this system. Edrophonium was found to be a more potent competitive inhibitor of AChE than either physostigmine or pyridostigmine, but a much weaker inihibitor of ChE than the latter two. Ambenonium behaves as a noncompetitive inhibitor of ChE; it is at least 10,000 times more effective on AChE and is 300 times more potent an inhibitor of AChE than is physostigmine. The use of edrophonium and ambenonium as selective inhibitors of AChE is suggested.
CYTOLOGICAL LOCALIZATION OF NORADRENALINE, MONOAMINE OXIDASE AND ACETYLCHOLINESTERASE IN SALIVARY GLANDS OF DOG
Authors: FUJIWARA MOTOHATSU , TANAKA CHIKAKO , HIKOSAKA HIROSHI , OKEGAWA TADAO
Published: April 2011

The cytological localization of noradrenaline, monoamine oxidase and acetylcholinesterase was examined in the salivary glands of the dog. The noradrenaline-fluorescent nerve fibers surrounded the secretory acini of the submaxillary, sublingual and parotid glands, but the density in the sublingual gland was much less than in the other two glands. Noradrenaline fluorescence was not seen in the cytoplasms of the acinar cells nor in the vicinity of the excretory ducts. Intense fluorescence was present outside the smooth muscle layer in various sized arteries. The distribution of monoamine oxidase activity was different from that of noradrenaline fluoresence. All the cells of the secretory acini and the excretory ducts were evenly stained. The distribution of fibers showing acetylcholinesterase activity was different from that of noradrenaline-fluorescent fibers only in that the former fibers were present around the excretory ducts as well. The composition of secretory cells in the submaxillary and sublingual glands is different in dogs and rats. However, the cytological localization of noradrenaline, monoamine oxidase and acetylcholinesterase activities in the salivary glands of the dog was essentially similar to those of the rat. It was concluded, therefore, that the autonomic nerve supply to the salivary glands does not correlate with the type of secretory cells.
Dissenting in reflective conversations: Critical components of doing action research
Authors: Bjørn Pernille , Boulus Nina
Published: April 2011

Reflective monitoring of research practices is essential. However, we often lack formal training in the practices of doing action research, and descriptions of actual inquiry practice are seldom included in publications. Our aim is to provide a glimpse of self-reflective practices based on our experience of enacting action research. Our engagement in a large action research project brought about many dilemmas and uncertainties related to our roles and interventions. We focus on these uncertainties as a way of opening the ‘black box’ of action research ‘in the making’. We conceptualize a methodological reflective approach that provides space for taking seriously uncertainties experienced in the field as these can be a catalyst for learning and sharpening our theoretical and empirical skills as action researchers. Through first-person inquiry, we investigate how our reflective conversations gradually evolved into second-person inquiry. We argue that enacting second-person reflective conversations renders alternative strategies for handling uncertainties through articulation of the tacit assumptions within particular empirical situations. Finally, we argue that reflective conversations should be understood and handled as iterative, open, and inquiring processes within a trust relationship, and should not be limited to any particular value schemes of action research.
ELECTRON MICROSCOPIC DEMONSTRATION OF DEHYDROGENASE ACTIVITY WITH A NEW OSMIOPHILIC DITETRAZOLIUM SALT (TC-NBT)
Authors: SELIGMAN ARNOLD M. , UENO HIROMI , MORIZONO YOSHIHISA , WASSERKRUG HANNAH L. , KATZOFF LIONEL , HANKER JACOB S.
Published: April 2011

Dissatisfaction with the degree of contrast produced by currently available tetrazolium salts in electron microscopy of the dehydrogenases stimulated us to design and prepare an osmiophilic ditetrazolium salt (TC-NBT)3 containing thiocarbamyl groups in innocuous positions of the nitro-BT molecule. The new reagent has the same favorable properties as nitro-BT for light microscopy and, in addition, is convertible to osmium black when tissue sections containing the diformazan are exposed to osmium tetroxide vapor. The stained tissue sections withstand dehydration and embedding in Araldite. Fresh and formalin-fixed rat heart, cut in thin slices without freezing, were stained for SDH and NADH2-D activity. In fresh heart, both dehydrogenases produced dense and thickened cristae in mitochondria that were active. Heterogeneity of mitochondria ris à vis dehydrogenase activity was confirmed. In fresh preparations stained mitochondria were contracted with narrowing of intercristal spaces as compared to inactive mitochondria. This was not the case in formalin-fixed heart. The dehydrogenase activity of individual mitochondria appeared to be an all-or-none phenomenon in these preliminary studies and the diformazan deposits were quite uniformly distributed along both surfaces of the cristae with encroachment on intracristal spaces in heavily stained preparations. The outer limiting membranes of the active mitochondria did not have diformazan deposits in contrast to the heavily stained cristae. Unreactive mitochondria revealed the usual contrast seen in control sections exposed to osmium tetroxide or in deep parts of the blocks to which TC-NBT did not penetrate.