bluuebunny/crossref_metadata_2025 huggingface dataset:
Lectin Histochemistry Reveals the Appearance of M-cells in Peyer's Patches of scid Mice After Syngeneic Normal Bone Marrow Transplantation
Authors: Sharma Ram , Schumacher Udo , Adam Elizabeth
Published: April 2011

Peyer's patches in the intestinal mucosa are characterized by the presence of several lymphatic follicles and interfollicular T-cell regions. Luminal antigens are transported across the intestinal epithelium to stimulate the Peyer's patch pre-B-cells in the follicles that proliferate and migrate to distant sites. Evidence suggests that antigen priming of B-lymphocytes is responsible for the number and location of Peyer's patches during postnatal life, but little is known about the histogenesis of Peyer's patches and their overlying membranous (M) cells. To examine whether T- and B-lymphocytes in Peyer's patches have an influence on M-cell generation, we studied the development of Peyer's patches and M-cells in severe combined immunodeficient (scid) mice reconstituted with bone marrow cells from normal syngeneic mice. Our experiments demonstrate that the donor bone marrow cells in the host scid mice repopulate to form single (primary) follicles and aggregates of lymphoid nodules, the Peyer's patches. Use of the lectins Anguilla anguilla (AAA) and Ulex europaeus I (UEA-I) as positive markers of mouse Peyer's patch M-cells revealed that M-cells develop in the dome epithelium overlying the single primary follicles and Peyer's patches of reconstituted scid mice. This experimental system therefore allows the study of the histogenesis of Peyer's patches and M-cells.
β-Dystroglycan Localization in the Photoreceptor and Müller cells in the Rat Retina Revealed by Immunoelectron Microscopy
Authors: Ueda Hideho , Gohdo Takashi , Ohno Shinichi
Published: April 2011

β-Dystroglycan (β-DG) is a dystrophin-associated glycoprotein that is expressed in skeletal muscle and other tissues. In the retina, dystrophin is present in the outer plexiform layer (OPL), where it is enriched under the photoreceptor cell membrane. In this study we determined the immunocytochemical localization of β-DG at both light and electron microscopic levels. β-DG immunoreactivity was detected at the inner limiting membrane, OPL, and around blood vessels. Immunoelectron microscopy detected β-DG immunoreactive products under the photoreceptor cell membrane, which are the same regions of dystrophin localization. In addition, β-DG was detected under the Müller cell membrane that is attached to the paravitreous or perivascular basement membrane. Our results suggest that β-DG may interact with dystrophin in photoreceptor membranes. However, β-DG-related interactions between Müller cells and basement membranes appear to be independent of dystrophin and raise the possibility that β-DG interacts with other molecules. We speculate that β-DG plays a role in maintaining the structural relationship between photoreceptor and bipolar cells or between Müller cells and basement membranes.
Pex mRNA Is Localized in Developing Mouse Osteoblasts and Odontoblasts
Authors: Frota Ruchon Andréa , Marcinkiewicz Mieczyslaw , Siegfried Géraldine , Tenenhouse Harriet S. , DesGroseillers Luc , Crine Philippe , Boileau Guy
Published: April 2011

Mutations in PEX, a phosphate-regulating gene with homology to endopeptidase on the X chromosome, were recently identified in patients with X-linked hypophosphatemia (XLH), an inherited disorder of phosphate homeostasis characterized by growth retardation and rachitic and osteomalacic bone disease. To understand the mechanism by which loss of PEX function elicits the mutant phenotype, a study of its mRNA localization and ontogenesis was undertaken. Using the reverse transcriptase-nested polymerase chain reaction (RT-nested PCR) with polyA+ RNA purified from mouse testis, a 337-bp Pex cDNA fragment was generated and cloned in the pCRII plasmid. The cDNA was used to generate sense and anti-sense Pex riboprobes for in situ hybridization (ISH) and Northern analysis. To survey a large number of different tissues, sagittal sections of embryos and newborn mice were examined. ISH showed the presence of Pex mRNA in osteoblasts and odontoblasts. Pex gene expression was detectable on Day 15 of embryonic development, which coincides with the beginning of intercellular matrix deposition in bones. Finally, Northern analysis of total RNA from calvariae and teeth of 3-day-old and adult mice showed that the abundance of the 7-kb Pex transcript is decreased in adult bones and in nongrowing teeth. The present study demonstrates that Pex mRNA is expressed in bones and teeth and suggests that this putative endopeptidase plays an important role in the development of these tissues.
Localization of Apoptotic Cells in the Human Epidermis by an In Situ DNA Nick End-labeling Method Using Confocal Reflectant Laser Microscopy
Authors: Ito Y. , Otsuki Y.
Published: April 2011

We describe an immunohistochemical method that allows the detection of apoptotic cells in human epidermis by use of confocal laser reflectance and antibody-immunogold-silver complexes. For this purpose, the site of free 3′-OH DNA ends was detected by the reflectance from heavy metal products (anti-digoxigenin antibody-immunogold-silver complexes) instead of 3, 3′ -diaminobenzidine (DAB) reaction products in the conventional in situ nick end-labeling of DNA strand breaks (ISEL) technique. Localization of double-stranded DNA was demonstrated by the autofluorescence of methyl green. The ISEL technique using confocal reflectant laser microscopy (CRLM) clearly showed the most intense reflectance in the nuclei of granular cells, in contrast to only a weaker reflectance in those of basal cells. On the other hand, the extent of autofluorescence of methyl green was significantly more intense in the nuclei of basal cells and showed a reciprocal relation to that of the reflectance. Therefore, granular cells were most prone to apoptosis and did not contain double-stranded DNA, as indicated by the lack of stainability with methyl green. In addition, this method demonstrating the simultaneous localization of both free 3′-OH DNA ends and double-stranded DNA proved to have a wide range of applications, including the study of other DNA autolytic processes.
Comparative Immunochemical Analyses of the Developmental Expression and Distribution of Ameloblastin and Amelogenin in Rat Incisors
Authors: Nanci A. , Zalzal S. , Lavoie P. , Kunikata M. , Chen W.-Y. , Krebsbach P.H. , Yamada Y. , Hammarström L. , Simmer J.P. , Fincham A.G. , Snead M.L. , Smith C.E.
Published: April 2011

SUMMARY Mineralized tissues are unique in using proteins to attract and organize calcium and phosphate ions into a structured mineral phase. A precise knowledge of the expression and extracellular distribution of matrix proteins is therefore very important in understanding their function. The purpose of this investigation was to obtain comparative information on the expression, intracellular and extracellular distribution, and dynamics of proteins representative of the two main classes of enamel matrix proteins. Amelogenins were visualized using an antibody and an mRNA probe prepared against the major alternatively spliced isoform in rodents, and nonamelogenins by antibodies and mRNA probes specific to one enamel protein referred to by three names: ameloblastin, amelin, and sheathlin. Qualitative and quantitative immunocytochemistry, in combination with immunoblotting and in situ hybridization, indicated a correlation between mRNA signal and sites of protein secretion for amelogenin, but not for ameloblastin, during the early presecretory and mid-to late maturation stages, during which mRNA signals were detected but no proteins appeared to be secreted. Extracellular amelogenin immunoreactivity was generally weak near secretory surfaces, increasing over a distance of about 1.25 μm to reach a level slightly above an amount expected if the protein were being deposited evenly across the enamel layer. Immunolabeling for ameloblastin showed an inverse pattern, with relatively more gold particles near secretory surfaces and much fewer deeper into the enamel layer. Administration of brefeldin A and cycloheximide to stop protein secretion revealed that the immunoblotting pattern of amelogenin was relatively stable, whereas ameloblastin broke down rapidly into lower molecular weight fragments. The distance from the cell surface at which immunolabeling for amelogenin stabilized generally corresponded to the point at which that for ameloblastin started to show a net reduction. These data suggest a correlation between the distribution of amelogenin and ameloblastin and that intact ameloblastin has a transient role in promoting/stabilizing crystal elongation.
Persistence of Biological Activity of Biotinylated Human Chorionic Gonadotropin and Its Use for Visualization of Rat Luteinizing Hormone Receptors in Tissue Sections
Authors: Paukku Tommi , Ahtiainen Petteri , Haavisto Anne-Maarit , Huhtaniemi Ilpo T.
Published: April 2011

Biotinylation of antibodies is an established method for producing systems for detection of antigens. We currently aim to develop liposomal targeting vectors for gene transfer into transgenic gonadal tumor cells expressing the luteinizing hormone (LH) receptor (R). We have biotinylated (B) human chorionic gonadotrophin (hCG) to obtain a selective targeting molecule to be attached to biotinylated liposomes via an avidin-streptavidin bridge. The biotinylation was performed by combining biotin isothiocyanate (BITC) and hCG in alkaline reaction buffer in a 100:1 (BITC:hCG) molar ratio. B-hCG maintained its ability to bind specifically to rat testicular membranes and was also bound to streptavidincoated polypropylene wells. cAMP production was induced in BLT-1 Leydig tumor cells in vitro after stimulation with B-hCG, as a sign of persistent bioactivity. Frozen sections of rat testicular and ovarian tissues and skeletal muscle were labeled by incubating for 2 hr at 37C with 10 ng/μl B-hCG. The binding was subsequently visualized by the avidin-biotin-peroxidase system, followed by silver enhancement of Ni-DAB staining. In rat testicular and ovarian sections, labeling was observed in structures known to strongly express the LH-R, i.e., Leydig cells, corpora lutea, and blood vessels. The labeling was blocked by preincubation with a 100-fold excess of the native hormone, and by injecting the rats SC with a high dose of hCG (1000 IU/kg) 48 hr before sacrifice. Skeletal muscle, used as negative control, was not labeled. These data demonstrate that the bioactivity of hCG is relatively well preserved after biotinylation. The biotinylated gonadotropin offers a new nonradioactive alternative for visualization of bioactive LH receptors in tissue sections.
Secreted Protein, Acidic and Rich in Cysteine (SPARC) and Thrombospondin in the Developing Follicle and Corpus Luteum of the Rat
Authors: Bagavandoss P. , Sage E. Helene , Vernon Robert B.
Published: April 2011

In adult mammals, growth of new vasculature from extant blood vessels (angiogenesis) is rare in the absence of pathology. However, nonpathogenic angiogenesis occurs in the cycling ovary when the avascular postovulatory follicle transforms into a highly vascularized corpus luteum (CL). To improve our understanding of molecular mechanisms that regulate nonpathogenic vascular growth, we characterized the expression of two secreted matricellular proteins associated with angiogenesis, SPARC and thrombospondin (TSP), in postovulatory preluteal follicles and CL of hormone-primed immature rats. By indirect immunofluorescence with specific antibodies, we found SPARC in the cytoplasm of granulosa cells and thecal cells of preluteal follicles, in connective tissue cells of the ovarian interstitium, and in the oocyte nucleus. Administration of a luteinizing stimulus (chorionic gonadotropin) increased the expression of SPARC in granulosa cells. TSP was prominent in the basement membranes of growing follicles. Many cells in the early vascularizing CL expressed both SPARC and TSP. Neovascularization of CL was accompanied by expression of SPARC in nascent vessels and concentration of TSP in central avascular areas. In mature CL, steroidogenic luteal cells expressed both SPARC and TSP. Luteal cells of regressing CL retained SPARC to a variable degree but did not express TSP. The observed changes in expression of SPARC and TSP during development of the CL support distinct roles for these matricellular proteins in nonpathological morphogenesis and angiogenesis.
A Rapid Procedure Suitable to Assess Quantitatively the Endocytosis of Colloidal Gold and Its Conjugates in Cultured Cells
Authors: Andreu Enrique José , Llano José Javier Martín de , Moreno Isabel , Knecht Erwin
Published: April 2011

We measured the endocytic uptake of low-density lipoproteins (LDLs) conjugated to colloidal gold in cultured cells, either by counting gold particles on electron micrographs or by inductively coupled plasma (ICP) mass spectrometry (MS). Both procedures are comparable but the latter requires a considerably shorter time and allows analysis of a much larger sample. In addition, ICP MS, compared to alternative radioactive or fluorescent procedures, offers the major advantage of using the same probe to quantify the endocytic uptake and to follow it by electron microscopy. Therefore, ICP MS analysis provides an easy, rapid, and sensitive quantification of endocytosis that complements the electron microscopic studies.
Immunoelectronmicroscopic Analysis of the Ligand-induced Internalization of the Somatostatin Receptor Subtype 2 in Cultured Human Glioma Cells
Authors: Krisch Brigitte , Feindt Janka , Mentlein Rolf
Published: April 2011

We analyzed the internalization of the receptor subtype 2 (sst2) for the neuropeptide somatostatin in glioma cells at the ultrastructural level using an antibody against an extracellular amino acid sequence. Intact cells derived from solid human gliomas or those of the human glioma cell line U343 were receptor-labeled (a) by classical gold immunocytochemistry using a 15-nm gold-labeled second antibody, (b) directly with the sst2 antibody adsorbed to 5-nm colloidal gold, and (c) with the physiological ligand somatostatin conjugated to 5-nm colloidal gold. The receptor was predominantly internalized via uncoated vesicles budding from the cell membrane but only rarely via coated pits, which has been mostly reported for G-protein-coupled, seven transmembrane-domain receptors. In the presence of ligand and sst2 antibody vesicles, tubule-like structures, and multivesicular bodies were labeled in superficial and in perinuclear portions of the cells within the first 30 min. Lysosomal labeling was observed after 30 min and especially after an hour of internalization time. This internalization route is also used to study the directly labeled sst2 antibody or the labeled ligand. However, the late endosomal compartment appears to be reached more rapidly in these latter experiments.
A Previously Unrecognized Site of Local Accumulation of Mononuclear Cells
Authors: Waltner-Romen Maria , Falkensammer Gerda , Rabl Walter , Wick Georg
Published: April 2011

In recent years our laboratory has developed an immunological hypothesis for the pathogenesis of atherosclerosis. We have shown that cellular and humoral immune reactions against heat shock proteins (Hsps) 60/65 expressed on the surface of stressed endothelial cells comprise the initial event in the pathogenesis of this disease. In the course of these studies, we also investigated normal, unaffected arteries for control purposes (carotid bifurcations from children aged 8 weeks to 10 years). This investigation led to the unexpected and previously unknown finding that mononuclear cells pre-exist in the intima at bifurcation sites. Our findings can be summarized as follows: Mononuclear cells are always found in the intima, primarily at sites subjected to major hemodynamic stress. Although the proportion of macrophages vs CD3+ T-cells differs, overall the latter clearly predominate. Most of the T-cells express the T-cell receptor (TCR)α/β, but TCRγ/δ cells are also present. We also identified dendritic cells and mast cells in the intima. Analogous to the mucosaassociated lymphoid tissue (MALT) we coined the designation “vascular-associated lymphoid tissue” (VALT) for these newly discovered cellular aggregates in the arterial intima.
Double Immunolabeling of Neuropeptides in the Human Hypothalamus as Analyzed by Confocal Laser Scanning Fluorescence Microscopy
Authors: Romijn Herms J. , van Uum Johanna F.M. , Breedijk Ingrid , Emmering Jasper , Radu Ioan , Pool Chris W.
Published: April 2011

The main goal of this study was to develop a better light microscopic procedure for quantitative study of the cellular co-localization of neuropeptides in adult human brain tissue. To reach this goal, we opted for a method (proved to be optimal on rat brain) in which sections were double immunolabeled with two different fluorophore-conjugated secondary antibodies and analyzed with a confocal laser scanning fluorescence microscope. One of our main problems faced was a strong autofluorescence of the sections, mainly caused by lipofuscin granules normally present in adult human brain tissue, which made any analysis of specific fluorescence impossible. This problem could be solved by staining the sections after immunolabeling with the dye Sudan Black B, which completely blocked this autofluorescence. The complete optimized procedure that we eventually developed can be summarized as follows. After a relatively short fixation time (6-14 days) in 4% freshly depolymerized paraformaldehyde, the resected brain tissue can best be stored in a 30% sucrose solution supplemented with 0.05% NaN3 at 4C. Stored under these conditions, cryosections from the tissue still reveal good histology and allow successful immunocytochemical staining after a period of 6 months. Double immunolabeling is done by incubating cryo- or paraffin sections in a mixture of two primary antibodies directed against the targeted antigens, followed by incubation with two different fluorophore-conjugated secondary antibodies. Amplification with a biotinylated secondary antibody followed by fluorophore-conjugated streptavidin is possible. Finally, the sections are stained with Sudan Black B, mounted in plain 80% Tris-buffered glycerol, and studied by confocal laser scanning fluorescence microscopy. Sections processed in this way are well suited for qualitative and quantitative analyses of co-localized neuropeptides in human brain tissue.
Visualization of Axonally Transported Horseradish Peroxidase Using Enhanced Immunocytochemical Detection: A Direct Comparison with the Tetramethylbenzidine Method
Authors: Romero Mario I. , Romero Martha A. , Smith George M.
Published: April 2011

Visualization of the neuronal tract tracer horseradish peroxidase (HRP) is commonly achieved through the histochemical detection of its enzymatic activity using 3,3′,5,5′-tetramethylbenzidine (TMB) as a chromogen. However, the TMB product is unstable and is incompatible with tissue processing methods that render the enzyme inactive, or when a combination of HRP tract tracing with neuronal phenotype identification is required. In this study we evaluated the applicability of the immunocytochemical detection method for horseradish peroxidase (HRP) visualization using an enhanced detection method based on the Elite ABC peroxidase amplification protocol. The results provide evidence for the immunocytochemical visualization of both anterograde and transganglionic HRP transport in the rat spinal cord. This immunocytochemical method not only showed similar sensitivity to the TMB protocol in detecting HRP-labeled motor neuron perikarya but provided enhanced resolution in the identification of individual neuronal fibers compared to the TMB method. Immunodetection of the HRP tracer also allowed its co-localization with specific neuronal markers using double immunofluorescence techniques. These results offer the first demonstration that sensitive identification of axonally transported HRP can be achieved by immunocytochemistry and provides further support for its use in HRP tract tracing studies.
Reduction of Lipofuscin-like Autofluorescence in Fluorescently Labeled Tissue
Authors: Schnell Stephen A. , Staines William A. , Wessendorf Martin W.
Published: April 2011

The fluorescent pigment lipofuscin accumulates with age in the cytoplasm of cells of the CNS. Because of its broad excitation and emission spectra, the presence of lipofuscin-like autofluorescence complicates the use of fluorescence microscopy (e.g., fluorescent retrograde tract tracing and fluorescence immunocytochemistry). In this study we examined several chemical treatments of tissue sections for their ability to reduce or eliminate lipofuscin-like autofluorescence without adversely affecting other fluorescent labels. We found that 1-10 mM CuSO4 in 50 mM ammonium acetate buffer (pH 5) or 1% Sudan Black B (SB) in 70% ethanol reduced or eliminated lipofuscin autofluorescence in sections of monkey, human, or rat neural tissue. These treatments also slightly reduced the intensity of immunofluorescent labeling and fluorescent retrograde tract tracers. However, the reduction of these fluorophores was far less dramatic than that for the lipofuscin-like compound. We conclude that treatment of tissue with CuSO4 or SB provides a reasonable compromise between reduction of lipofuscin-like fluorescence and maintenance of specific fluorescent labels.
The Multiple Disability Implicit Association Test
Authors: Vaughn E. Daly , Thomas Adrian , Doyle Andrea L.
Published: April 2011

The authors report the psychometric properties of the Multiple Disability Implicit Association Test (MDIAT) using the updated implementation procedure and scoring algorithm provided by Greenwald and colleagues. Undergraduate students ( N = 249) completed the four disability condition IATs composing the MDIAT in addition to several self-report scales assessing constructs of interest (e.g., explicit attitude toward persons with disabilities, socially desirable responding, cognitive ability). Consistent with previous research, MDIAT effect scores indicated implicit preference for persons without a disability relative to persons with a disability for four disability conditions and a composite score. MDIAT scores were not related to socially desirable responding or cognitive ability. However, “practice” effects previously found under the conventional scoring algorithm were replicated with the improved scoring algorithm. Finally, scores on the MDIAT were not related to an explicit measure of attitude, the Interaction with Disabled Persons Scale. Although the modifications and new scoring algorithm attenuated some problems found in the previous administrations of the MDIAT (e.g., confounds from socially desirable responding), the practice effect persisted. Until more is known about the underlying processes responsible for the decline in IAT effect scores across administrations, those using the MDIAT should interpret only the composite effect score.
The Mainstream Is Not Electable: When Vision Triumphs Over Representativeness in Leader Emergence and Effectiveness
Authors: Halevy Nir , Berson Yair , Galinsky Adam D.
Published: April 2011

Theories of visionary leadership propose that groups bestow leadership on exceptional group members. In contrast, social identity perspectives claim that leadership arises, in part, from a person’s ability to be seen as representative of the group. Integrating these perspectives, the authors propose that effective leaders often share group members’ perspectives concerning the present, yet offer a unique and compelling vision for the group’s future. In addition, although intergroup contexts may increase the value of representativeness, the authors predict that vision dominates representativeness in single-group situations characterized by high levels of collective stress (e.g., a natural disaster). Five studies demonstrated that visionary leaders (those who offer novel solutions to their group’s predicament) attract more followers, promote group identification and intrinsic motivation, mobilize collective action, and effectively regulate group members’ emotions and reactions to crises compared to representative leaders. The authors discuss when, why, and how vision triumphs over representativeness in leader emergence and effectiveness.
A Change Will Do Us Good: Threats Diminish Typical Preferences for Male Leaders
Authors: Brown Elizabeth R. , Diekman Amanda B. , Schneider Monica C.
Published: April 2011

The current research explores role congruity processes from a new vantage point by investigating how the need for change might shift gender-based leadership preferences. According to role congruity theory, favorability toward leaders results from alignment between what is desired in a leadership role and the characteristics stereotypically ascribed to the leader. Generally speaking, these processes lead to baseline preferences for male over female leaders. In this research, the authors propose that a shift in gender-based leadership preferences will emerge under conditions of threat. Because the psychological experience of threat signals a need for change, individuals will favor candidates who represent new directions in leadership rather than consistency with past directions in leadership. Specifically, they find that threat evokes an implicit preference for change over stability (Experiment 1) and gender stereotypes align women with change but men with stability (Experiments 2a and 2b). Consequently, the typical preference for male leaders is diminished, or even reversed, under threat (Experiments 3 and 4). Moreover, the shift away from typical gender-based leadership preferences occurs especially among individuals who highly legitimize the sociopolitical system (Experiment 4), suggesting that these preference shifts might serve to protect the underlying system.
Lessons of History Part II
Authors: Bunkers Sandra Schmidt
Published: April 2011

This column is an introduction to a manuscript depicting a story of historical change in nursing.
Hidden Treasures in Co-shaping Community Environment
Authors: Condon Barbara Backer
Published: April 2011

In this column the author focuses on the concepts of growing community, cultivating trust, and keeping passion as part of co-shaping community. These concepts surfaced in the story of Mae Gonzalez, a 90 year-old nurse who is still active in her nursing practice. Her story, intermingled throughout this column, demonstrates the importance of historical consciousness to current nursing. Three of the processes from Parse’s description of community as well as her concepts of true presence and human dignity are highlighted as Mae shared her treasures of growing community, cultivating trust, and keeping passion.
Applicant Quality: Exploring the Differences Between Organizational and Third-Party Websites
Authors: Talmage Craig
Published: April 2011

Few studies have assessed the differences between organizational and third-party websites, especially with regards to applicant quality. Organizational websites are hypothesized to bring in higher quality applicants than third-party websites because of the amount of organization-specific information they contain. This study assesses perceived differences between web recruitment sources in applicant quality using a convenience sample of resumes from job applicants from a midsize company. Resumes were rated by human resource personnel in terms of applicant quality. The organizational website contained higher quality applicants on average compared to four third-party website sources.
CYTOCHEMICAL OBSERVATIONS ON CHICKEN MONOCYTES MACROPHAGES AND GIANT CELLS IN TISSUE CULTURE
Authors: WEISS LEON PAUL , FAWCETT DON W.
Published: April 2011

Cultures of leukocytes from chicken blood grown in fluid medium in roller flasks were stained for lipids, carbohydrates, phosphatase, esterase, and succinic dehydrogenase in order to study the cytochemical changes associated with the transformation of monocytes to macrophages, epithelioid cells and multinucleate giant cells.

The great variability in cell form observed by previous investigators in such cultures was confirmed. The cells were usually round but not uncommonly were stellate or fusiform, sometimes bearing a striking similarity to fibroblasts, at other times resembling sheets of mesothelium. The factors determining their size and shape were obscure, but low pH seemed to favor the formation of giant cells.

Monocytes of chicken blood were ordinarily devoid of stainable lipid, carbohydrate and enzymes. Macrophages and epithelioid cells which developed from them were negative for esterase, succinic dehydrogenase and alkaline phosphatase but gave a strong reaction for acid phosphatase, localized in their enlarged centrosphere or Golgi zone. This region also contained a saliva-resistant, periodic acid-Schiff positive material, probably a carbohydrate-protein complex and its relation to the enzyme acid phosphatase has been considered. Numerous small lipid droplets were disposed around the periphery of the Golgi zone. Giant cells occasionally contained glycogen.

The acquisition of acid phosphatase by monocytes during their transformation to macrophages appeared to be related to their enhanced phagocytic activity.
STUDIES IN HISTOCHEMISTRY: XXVII. THE DETERMINATION OF L-ASCORBIC ACID, AND DEHYDRO-L-ASCORBIC ACID PLUS DIKETO-L-GULONIC ACID IN MICROGRAM QUANTITIES OF TISSUE
Authors: GLICK DAVID , ALPERT MORTON , STECKLEIN HELEN R.
Published: April 2011

The determination of l-ascorbic, and dehydro-l-ascorbic plus diketo-l-gulonic acids in microgram quantities of tissue was described. The procedure used is an adaptation to the micro scale of macro methods based on spectrophotometric measurements of reactions involving 2,6-dichlorophenol indophenol and 2,4-dinitrophenylhydrazine, and it is well suited to serial analyses on large numbers of samples because of its relatively great simplicity and speed. The standard deviation in the ascorbic acid method is about 2% corresponding to about 3 mµg. of ascorbic acid for the equipment used. With pipettes and cuvettes of smaller capacity a reduction in the latter value by about one-third can be obtained without inconvenience.
A CHEMICAL AND HISTOCHEMICAL INVESTIGATION OF GLYCOGEN IN RAT LIVER AND PALATE FOLLOWING TREATMENT WITH VARIOUS FIXATIVES AND ETHYLENEDIAMINE TETRAACETIC ACID
Authors: TROTT J. R. , GORENSTEIN S. L. , PEIKOFF M. D.
Published: April 2011

The loss of glycogen after fixation in acetic acid-alcohol-formalin as compared to immediate determination was found by chemical analysis to be from 5.2% to 13.8%. No appreciable difference could be seen histochemically.

Chemical analysis of the glycogen content of liver fixed in acetic acid-alcohol-formalin and then treated in ethylenediamine tetracetic acid showed a total drop of 20.7 to 33.7%. Glycogen could still be adequately shown histochemically.

Sections taken from liver fixed in 1% periodic acid in 10% formalin show greater amounts of glycogen, than those taken from acetic-acid-alcohol-formalin fixed tissue.

After fixation in 1% periodic acid in 10% formalin and following treatment with ethylenediamine tetraacetic acid, very little glycogen could be demonstrated histochemically.

In the palate, no glycogen could be demonstrated after fixation in acetic acid-alcohol-formalin, whereas quite heavy amounts could be seen in the prickle cell layers of the epithelium following fixation in 1% periodic acid in 10% formalin.

Neither acetic acid-alcohol-formalin nor 1% periodic acid in 10% formalin are satisfactory fixatives if the tissue is to be further decalcified in ethylenediamine tetraacetic acid, if there are only small amounts of glycogen present.

No adequate method of retaining glycogen in small amounts in tissue requiring decalcification can be suggested.
HISTOCHEMICAL DEMONSTRATION OF A GAMMA-GLUTAMYL TRANSPEPTIDASE-LIKE ACTIVITY
Authors: GLENNER G. G. , FOLK J. E. , MCMILLAN P. J.
Published: April 2011

A histochemical method for the demonstration of a γ-glutamyl transpeptidase-like enzymic activity using N(γ-glutamyl)β-naphthylamide as substrate in a simultaneous diazonium coupling reaction has revealed many tissue localizations distinct from those noted with other peptidase substrates. The biochemical and histochemical evidence presented suggests that at the pH of the histochemical reaction both a transpeptidase and hydrolytic reaction mediated by the same enzyme occurs and, as indicated by histochemical inhibitor studies, the characteristics of the activity appear to be identical with those of γ-glutamyl transpeptidase as defined biochemically.
A HISTOCHEMICAL STUDY OF THE INCORPORATION OF GLUCOSE C14 INTO GLYCOGEN BY LIVER SLICES IN VITRO
Authors: KUGLER J. H. , WILKINSON W. J. C.
Published: April 2011

The in vitro formation of glycogen from d-glucose-C14 has been studied by histochemical and autoradiographic techniques. The incorporation of the radioactive glucose into acid soluble glycogen lends support to the suggestion that this is the only glycogen fraction that is demonstrated histochemically. These experiments also confirm the validity of the histochemical tests for glycogen.
Predictors of Depressive Symptoms in Parents of Chronically Ill Children Admitted to the Pediatric Intensive Care Unit
Authors: Fauman Karen R. , Pituch Kenneth J. , Han Yong Y. , Niedner Matthew F. , Reske Janella , LeVine Ann Marie
Published: April 2011

Objective: To identify factors in the pediatric intensive care unit (PICU) patient population that may result in increased risk of depressive symptoms in their parents. Design: Six-month, prospective, observational study in a tertiary-level PICU on parents of chronically ill children admitted to PICU. Parents were assessed by background questionnaire and standardized depression scale. Results: Data was compared to various markers such as child’s diagnosis, admission reason, palliative care diagnosis type (ACT code), and course/length of disease. Incidence of depressive symptoms in parents was inversely correlated with duration of child’s chronic illness. Parents of children admitted for planned postoperative management were more likely to report depressive symptoms compared to parents of children admitted for acute changes in health. Conclusion: Parents of certain chronically ill children may benefit from routine screening for depression.
My Child is Diagnosed With Asthma, Now What?
Authors: Stepney Cesalie , Kane Katelyn , Bruzzese Jean-Marie
Published: April 2011

Pediatric asthma is often undiagnosed, and therefore untreated. It negatively impacts children’s functioning, including school attendance and performance, as well as quality of life. Schoolwide screening for asthma is becoming increasingly common, making identification of possible asthma particularly relevant for school nurses. Nurses may need to help parents cope with the new diagnosis, and teach them skills to manage the illness. The aim of this article is to present a three-phase model of how parents cope with a newly diagnosed pediatric chronic illness. Using asthma as an example, we describe these phases (Emotional Crisis, Facing Reality, and Reclaiming Life), illustrate how parents progress through the phases, and discuss situations associated with possible regression. Next, we offer strategies framed around a theory of asthma self-management to assist school nurses and other medical providers to motivate parents to develop successful disease management skills.
Validation of the Social Appearance Anxiety Scale: Factor, Convergent, and Divergent Validity
Authors: Levinson Cheri A. , Rodebaugh Thomas L.
Published: April 2011

The Social Appearance Anxiety Scale (SAAS) was created to assess fear of overall appearance evaluation. Initial psychometric work indicated that the measure had a single-factor structure and exhibited excellent internal consistency, test—retest reliability, and convergent validity. In the current study, the authors further examined the factor, convergent, and divergent validity of the SAAS in two samples of undergraduates. In Study 1 ( N = 323), the authors tested the factor structure, convergent, and divergent validity of the SAAS with measures of the Big Five personality traits, negative affect, fear of negative evaluation, and social interaction anxiety. In Study 2 ( N = 118), participants completed a body evaluation that included measurements of height, weight, and body fat content. The SAAS exhibited excellent convergent and divergent validity with self-report measures (i.e., self-esteem, trait anxiety, ethnic identity, and sympathy), predicted state anxiety experienced during the body evaluation, and predicted body fat content. In both studies, results confirmed a single-factor structure as the best fit to the data. These results lend additional support for the use of the SAAS as a valid measure of social appearance anxiety.
Kindergarten Reading Skill Level and Change as Risk Factors for Chronic Problem Behavior
Authors: McIntosh Kent , Sadler Carol , Brown Jacqueline A.
Published: April 2011

In this study, the authors explored the effect of prereading skills at the start of kindergarten and change in skills during kindergarten on response to Tier I (universal) Schoolwide Positive Behavior Support in Grade 5. A longitudinal data set of 473 students, including Dynamic Indicators of Basic Early Literacy Skills measures at the start, middle, and end of kindergarten and office discipline referrals in Grade 5, was used to determine whether reading skills at school entry or change in reading skills over the course of kindergarten were more predictive of chronic problem behavior in Grade 5. Results of logistic regression analyses indicated that low initial phonological awareness predicted problem behavior, but including skill growth in the model resulted in significantly improved and more accurate prediction. Results are discussed in terms of early screening and intervention and reducing risk for problem behavior through quality Tier I reading instruction in kindergarten.
STUDIES IN HISTOCHEMISTRY: LXX. QUANTITATIVE HISTOLOGICAL DISTRIBUTION OF TOTAL LIPID IN THE ADRENAL GLAND OF THE RAT MEASURED BY X-RAY MICRORADIOGRAPHY
Authors: HIRAOKA TOSISUKE , GLICK DAVID
Published: April 2011

Change in total dry mass following extraction with petroleums ether, as measured by X-ray absorption, was used to determine lipid in each histological zone of dried fresh-frozen microtome cross-sections of adrenal glands of male albino rats. Although the area of measurement was 1.1 µ2, and the data were given in units of 10–12 gm/µ2, the differences in dry mass per unit area in this material were not great enough to permit useful analytical resolution below the histological level. The data were in general agreement with those obtained from differences in mass by direct weighing of a series of whole dried fresh-frozen microtome sections which had been cut to segregate the various zones. The particular advantage of the X-ray method is that it permits determination of the lipid in situ in each of the histological zones of the gland in a single microtome cross-section, and the pair of microradiograms taken, before and after extraction, provides a permanent record from which measurements can be obtained subsequently at any time. Mean values found for total lipid, as percent of total dry mass, in adrenal zones of untreated rats were 12, capsule; 17, glomerulosa; 36, fasciculata; 36, reticularis; and 19, medulla.
THE LOCALIZATION AND SIGNIFICANCE OF ARGININE AND CITRULLINE IN PROTEINS OF THE HAIR FOLLICLE
Authors: ROGERS G. E.
Published: April 2011

The Sakaguchi reaction for arginine and the Ehrlich reagent for citrulline, which was modified to lessen tissue swelling, have been used to localize these amino acids in the inner sheath and the medulla layers in sections of actively growing hair follicles of the rat. The trichohyalin droplets are rich in arginine but gave no detectable reaction for citrulline. Conversely, the hardened substance of the inner sheath and of the medulla gave a relatively weak reaction for arginine but an intense one for citrulline. The presence of high concentrations of citrulline in the proteins of the hardened inner sheath and medulla has only recently been reported from chemical studies.

The results support biochemical and electron microscope studies indicating a conversion of arginine residues into citrulline residues without protein breakdown, concomitant with the morphological transformation of trichohyalin droplets into the hardened fibrous protein of the inner sheath or the hardened amorphous material of the medulla. Other possible biochemical functions for the inner sheath and the medulla are also discussed.
THE DISTRIBUTION OF LACTATE DEHYDROGENASE ISOZYMES IN HUMAN SKELETAL MUSCLE FIBERS
Authors: VAN WIJHE M. , BLANCHAER M. C. , ST. GEORGE-STUBBS S.
Published: April 2011

A study of the distribution of lactate dehydrogenase isozymes in single fibers from normal human skeletal muscle is presented. The fibers were classified into red, intermediate and white types on histochemical grounds and their lactate dehydrogenase isozyme content assessed by electrophoretic separation in veronal buffered agar. The results generally agreed with previous homogenate studies on animal skeletal muscle, in that the white fibers contained almost exclusively isozymes IV and V, whereas red fibers were rich in isozymes I, II and III, but IV and V also appeared indigenous to these fibers. The intermediate fibers had an isozyme pattern combining the features of red and white fibers. The metabolic implications of these findings are discussed.
COMPLEX FORMATION BETWEEN HEAVY METALS AND THIOLACETIC ACID IN ELECTRON MICROSCOPIC HISTOCHEMISTRY
Authors: HAUGAARD NIELS , HORN ROBERT S. , KOELLE GEORGE B.
Published: April 2011

Evidence for the formations of complexes of aurous and lead ions with thiolacetic acid was obtained by two methods. (1) It was demonstrated that the addition of the metal ions to solutions of thiolacetic acid at pH 6.2 caused a bathochromic shift of the ultraviolet spectrum of this compound characteristic of those observed on combination of metal ions with an organic resonator. The experiments indicated that aurous ions form a coordinate complex with thiolacetic acid containing one metal ion and two thiolacetate residues. (2) It was observed that aurous and lead ions inhibited the reaction of thiolacetic acid with N-ethylmaleimide. Complex formation of the heavy metals with thiolacetic acid may be of importance in preventing their binding at nonspecific sites and in facilitating their penetration into tissue sections in the adaptation for electron microscopy of the thiolacetic acid method for cholinesterases.
HISTOCHEMICAL ACYLATION OF ALDEHYDES PRODUCED BY PERIODIC ACID OXIDATION
Authors: LILLIE R. D.
Published: April 2011

Aldehydes produced in tissue sections by periodic acid oxidation are readily acetylated or benzoylated so as to weaken or completely prevent the Schiff and other chromogenic reactions. The reactivity of acylated aldehydes to Schiff reagent is promptly restored by saponification in alcoholic potassium hydroxide solution (10-20 min, 1% KOH, 70% ethanol). Benzoylation with 5-10% benzoyl chloride in pyridine gave the promptest, most complete and consistent aldehyde blockade, which was somewhat inferior on addition of 0.5% sulfuric acid. Pyridine acetic anhydride mixtures, 25, 40 and 50% gave partial to complete blockade. Addition of 0.25-0.5% sulfuric acid did not give consistent effects. Acetic anhydride at 60°C gave partial blockade at ½-5 hr, on addition of 0.01-0.25% sulfuric acid total or subtotal blockade was achieved. Acetylation in alcohol gave inferior resutlts. Use of 25% acetic anhydride ims glacial acetic acid gave inferior results; addition of 0.25% sulfuric acid produced total to subtotal blockade at 4-5 hr. Glacial acetic acid was without appreciable blockade effect. Sulfation in 10% and 25% H2SO4/glacial acetic acid failed to blockade aldehydes. Experiments with the peracetic acid Schiff reaction for ethylene groups indicate that some proportion of enol monobenzoate may be formed, and that with the various acetylation techniques a smaller or negligible proportion of enol acetate is formed. Acetic anhydride with 0.25% H2S04 at 60° appears to form only aldehyde diacetate. Experiments with the bromination silver techniques did not give satisfactory results.
On marketing theory and service-dominant logic: Connecting some dots
Authors: Vargo Stephen L.
Published: April 2011

This article serves as an introduction to a special section on ‘Extending Service-dominant Logic’, which includes articles developed from manuscripts presented at the Forum on Markets and Marketing held in Sydney, Australia. This special section is, in turn, part of a combined ‘special issue’, with related articles published in the Australasian Marketing Journal, the European Journal of Marketing, and the Journal of Macromarketing. Together with the articles in these journals, the four articles presented here move S-D logic closer to a theory of the market and marketing by further connecting conceptual dots and exploring the role of theory in relation to S-D logic, and S-D logic in relation to theoretical orientations.
CYTOCHEMICAL DEMONSTRATION OF β-GLUCURONIDASE ACTIVITY IN BLOOD AND BONE MARROW CELLS
Authors: LORBACHER PETER , YAM LUNG T. , MITUS WLADYSLAW J.
Published: April 2011

A method for the cytochemical demonstration of β-glucuronidase activity in blood and bone marrow is described using naphthol AS-BI-β-d-glucosiduronic acid as substrate and hexazotized pararosanilin as coupling reagent. β-Glucuronidase was found to be present in most of the cells of blood and marrow. The reaction was strongly positive in macrophages, monocytes, plasma cells and some of the lymphocytes. It was moderately positive in granulocytes and thrombocytes and weakly positive in cells of the erythroid series. β-Glucuronidase is known to be a lysosomal enzyme, but certain discrepancies exist between the content of this enzyme and the number of lysosomal structures as seen electron microscopically. It appears that not all of the lysosomes contain β-glucuronidase and that sites of β-glucuronidase other than lysosomal exist in certain cell lines, i.e., the lymphocytes and the plasma cells. For this reason, it is suggested that cytochemical investigations of lysosomes should always include the simultaneous demonstration of different lysosomal hydrolases.
DISTRIBUTION OF ESTERASES IN THE MYONEURAL JUNCTION OF THE STRIATED MUSCLE OF THE RAT
Authors: ERÄNKÖ O. , TERÄVÄINEN H.
Published: April 2011

Distribution of esterases in the myoneural junction of the striated muscle of the rat was studied using acetylthiocholine, butyrylthiochobine and α-naphthyl acetate as substrates, together with selective inhibitors.

Acetylcholinesterase activity was observed in the peripheral complex of synaptic folds. Nonspecific cholinesterase was detected in the peripheral complex of synaptic folds and the teloglia with approximately equal activities. Nonspecific esterase activity, present in tissues incubated with eserine, was marked in the terminal axon and was also present in the teloglia and synaptic folds.
THIN LAYER CHROMATOGRAPHIC STUDY OF LIPID EXTRACTION FROM CRYOSTAT SECTIONS OF RAT HYPOTHALAMUS BY SOME FIXATIVES
Authors: ROOZEMOND R. C.
Published: April 2011

Lipids of different classes are extracted considerably from cryostat sections of rat hypothalamus by water. The use of 4% formol, but especially of 1% calcium chloride, and a mixture of 4% formol and 1% calcium chloride decreases the amount of lipids extracted. The fact that even the latter fixative extracts significant amounts of lipids may explain the failure of the Baker acid hematein test in demonstrating the dark staining cells in the supraoptic and paraventricular nuclei when applied to fresh frozen sections of rat hypothalamus.