bluuebunny/crossref_metadata_2025 huggingface dataset:
Differential Expression of Gap Junction Proteins Connexin26, 32, and 43 in Normal and Crush-injured Rat Sciatic Nerves: Close Relationship Between Connexin43 and Occludin in the Perineurium
Authors: Nagaoka Takanori , Oyamada Masahito , Takamatsu Tetsuro , Okajima Seiichiro , Takamatsu Tetsuro
Published: April 2011

We immunohistochemically and morphometrically examined the expression of gap junction protein connexin (Cx) in normal and crush-injured rat sciatic nerves using confocal laser scanning microscopy. Cx26 was localized in the perineurium and Cx43 was present in the perineurium and the epineurium, whereas Cx32 was confined to the paranodal regions of the nodes of Ranvier. Double labeling for connexins and laminin revealed that Cx43 was localized in multiple layers of the perineurium, whereas Cx26 was confined to the innermost layer. Double labeling for connexins and a tight junction protein, occludin, showed that occludin frequently coexisted with Cx43 but existed separately from Cx26 in the perineurium. After crush injury, the pattern of normal Cx32 expression was initially lost but recovered, whereas Cx43 rapidly appeared in the endoneurium and its expression was subsequently attenuated. Although crush injury produced no apparent alteration in Cx43 and occludin in the perineurium, a rapid increase and a subsequent decrease in the frequency of Cx26-positive spots during nerve regeneration were shown by morphometric analysis. These results indicate that Cx26, Cx32, and Cx43 are expressed differently in various types of cells in peripheral nerves and that their expressions are differentially regulated after injury. The expression of connexins and occludin in the perineurium suggests that perineurial cells are not uniform in type and that the regulation of gap junctions and tight junctions is closely related in the perineurium.
Immunoglobulin and Enzyme-conjugated Dextran Polymers Enhance u-PAR Staining Intensity of Carcinoma Cells in Peripheral Blood Smears
Authors: Werther Kim , Normark Michel , Hansen Birgit Fischer , Nielsen Hans Jørgen
Published: April 2011

The presence of disseminated carcinoma cells in bone marrow and peripheral blood has prognostic importance in patients with carcinomas. Much evidence indicates that dissemination of tumor cells may depend on activation of a variety of degradative enzymes. A strong positive correlation has been shown between the expression of tumor cell proteases and tumor invasion. Therefore, phenotypic characterization of disseminated carcinoma cells for expression of protease activators might define the invasive potential of the cells. We present an immunocytochemically enhanced staining method that allows phenotyping of disseminated carcinoma cells in bone marrow and peripheral blood smears. In the first step, the cells were incubated with antibodies against urokinase plasminogen activator receptor (u-PAR) and subsequently with secondary antibodies conjugated to peroxidase-labeled dextran polymers. A brown color reaction was developed with diaminobenzidine as chromogen. In the second step, the cells were incubated with alkaline phosphatase-conjugated murine monoclonal antibodies against a common cytokeratin epitope and a red color reaction was developed with new fuchsin as substrate. This method allows simultaneous and unambiguous immunolabeling of intracellular cytokeratin and of u-PAR intracellularly and on the surface of carcinoma cells. This novel approach can be used for detection and phenotyping of carcinoma cells in blood smears for u-PAR or, presumably, for any other heterogeneously expressed antigen on the surface of the detected cells.
Gastric Amylin Expression: Cellular Identity and Lack of Requirement for the Homeobox Protein PDX-1. A Study in Normal and PDX-1-Deficient Animals with a Cautionary Note on Antiserum Evaluation
Authors: Tingstedt Jens-Erik , Edlund Helena , Madsen Ole D. , Larsson Lars-Inge
Published: April 2011

The gene encoding amylin is implicated in the generation of amyloid in the islets of Langerhans of diabetics and is believed to be regulated by the homeodomain transcription factor PDX-1. Although gastric mucosa also produces amylin, studies on its cellular site of production have yielded highly divergent results, localizing this peptide to either gastrin, serotonin, or somatostatin cells or to combinations thereof. Using region-specific amylin antisera in combination with reverse transcriptase-polymerase chain reaction, we now document that the majority of cells expressing amylin correspond to somatostatin cells. Only a small subpopulation of gastrin cells contained immunoreactive amylin. Studies of PDX-1-deficient mice, which fail to develop gastrin cells while possessing normal numbers of somatostatin cells, revealed no detectable change in gastric amylin expression. These data show that neither normal gastrin cell development nor PDX-1 expression is needed for gastric amylin expression. (J Histochem Cytochem 47:973–980, 1999)
Immunohistochemical Detection of α1E Voltage-gated Ca2+ Channel Isoforms in Cerebellum, INS-1 Cells, and Neuroendocrine Cells of the Digestive System
Authors: Grabsch Heike , Pereverzev Alexei , Weiergräber Marco , Schramm Martin , Henry Margit , Vajna Rolf , Beattie Ruth E. , Volsen Steve G. , Klöckner Udo , Hescheler Jürgen , Schneider Toni
Published: April 2011

Polyclonal antibodies were raised against a common and a specific epitope present only in longer α1E isoforms of voltage-gated Ca2+ channels, yielding an “anti-E-com” and an “anti-E-spec” serum, respectively. The specificity of both sera was established by immunocytochemistry and immunoblotting using stably transfected HEK-293 cells or membrane proteins derived from them. Cells from the insulinoma cell line INS-1, tissue sections from cerebellum, and representative regions of gastrointestinal tract were stained immunocytochemically. INS-1 cells expressed an α1E splice variant with a longer carboxy terminus, the so-called α1Ee isoform. Similarily, in rat cerebellum, which was used as a reference system, the anti-E-spec serum stained somata and dendrites of Purkinje cells. Only faint staining was seen throughout the cerebellar granule cell layer. After prolonged incubation times, neurons of the molecular layer were stained by anti-E-com, suggesting that a shorter α1E isoform is expressed at a lower protein density. In human gastrointestinal tract, endocrine cells of the antral mucosa (stomach), small and large intestine, and islets of Langerhans were stained by the anti-E-spec serum. In addition, staining by the anti-E-spec serum was observed in Paneth cells and in the smooth muscle cell layer of the lamina muscularis mucosae. We conclude that the longer α1Ee isoform is expressed in neuroendocrine cells of the digestive system and that, in pancreas, α1Ee expression is restricted to the neuroendocrine part, the islets of Langerhans. α1E therefore appears to be a common voltage-gated Ca2+ channel linked to neuroendocrine and related systems of the body. (J Histochem Cytochem 47:981–993, 1999)
Intracellular Distribution of a Biotin-labeled Ganglioside, GM1, by Immunoelectron Microscopy After Endocytosis in Fibroblasts
Authors: Möbius Wiebke , Herzog Volker , Sandhoff Konrad , Schwarzmann Günter
Published: April 2011

A radioactive and biotin-labeled analogue of GM1 (biotin-GM1) was synthesized which enabled us to analyze its intracellular distribution in the compartments of the endocytic route by electron microscopic immunocytochemistry using thin sections of human skin fibroblasts labeled with gold-conjugated antibiotin antibodies. Metabolic studies with the biotin-GM1 showed its partial degradation to the corresponding GM2 and GM3 derivatives. Further degradation was inhibited by the biotin residue. The distribution of biotin-GM1 after uptake by cells was studied by postembedding labeling techniques. On the plasma membrane the biotin-GM1 was detectable in the form of patches (0.1 μm in diameter), in caveola-like structures and, to a much lesser extent, in coated pits or vesicles. During endocytic uptake, the biotin-GM1 became detectable in organelles identified as late endosomes and lysosomes. The intracellular distribution of the biotin-GM1 was compared to the localization of the EGF receptor in EGF-stimulated fibroblasts. Both the biotin-GM1 and the EGF receptor were transported to intraendosomal and intralysosomal membranes, indicating that both membrane constituents follow the same pathway of endocytosis. Our observations show that biotin-GM1 can be successfully incorporated into the plasma membrane and be used as a tool for morphological detection of its pathway to lysosomes. (J Histochem Cytochem 47:1005–1014, 1999)
Ultrastructural Distribution of 36-kD Microfibril-associated Glycoprotein (MAGP-36) in Human and Bovine Tissues
Authors: Toyoshima Tetsuhiko , Yamashita Kayoko , Furuichi Hiromi , Shishibori Tsuyoshi , Itano Toshifumi , Kobayashi Ryoji
Published: April 2011

We observed the ultrastructural distribution of MAGP-36 by immunoelectron microscopy in human and bovine tissues. MAGP-36 was present in microfibrils associated with tropoelastin in skin, aorta, and spleen. It was not detected in microfibrils from the ocular zonule and kidney mesangium that were not associated with tropoelastin. In skin, MAGP-36 was present in both early immature elastic fibers and mature elastic fibers. In mature elastic fibers, MAGP-36 was localized around amorphous elastic cores at the elastin-microfibril interface and in electron-dense bundles. Localization of MAGP-36 in elastic fibers coincided with the distribution of lysyl oxidase, an enzyme that plays a pivotal role in the deposition of tropoelastin. These findings suggest that MAGP-36 may be involved in elastogenesis. (J Histochem Cytochem 47:1049–1056, 1999)
Immunocytochemical and In Situ Hybridization Studies of Gastrin After Cisplatin Treatment
Authors: Wang Ying , Aggarwal Surinder K. , Painter Cory L.
Published: April 2011

Cisplatin treatment (9 mg/kg) causes bloating of the stomach, an increase in gastric acid, and ulceration in rats. Gastrin, a gut peptide, plays an important role in regulating gastric acid production. To study the role of gastrin in this increased gastric acid production after cisplatin treatment, male Wistar rats (100–150 g) were treated with cisplatin (9 mg/kg) in five divided doses over 5 consecutive days. The rats were sacrificed 1, 6, 10, or 15 days after the last treatment. As measured by immunocytochemistry, in situ hybridization, Northern blot, and dot-blot techniques, gastrin was found to be below detectable limits just 1 day after cisplatin treatment. However, 10–15 days after the last injection, the levels for both gastrin and its mRNA gradually recovered to normal. Northern blot studies showed that decreased somatostatin mRNA parallels the changes of gastrin and its mRNA. These results suggest that after cisplatin treatment the increased gastric acid production in rat stomach is independent of gastrin. This decrease of gastrin production is not under the influence of somatostatin, which also decreased after cisplatin treatment. (J Histochem Cytochem 47:1057–1062, 1999)
Ultrastructural Localization of Epithelial Mucin Core Proteins in Colorectal Tissues
Authors: Winterford Clay M. , Walsh Michael D. , Leggett Barbara A. , Jass Jeremy R.
Published: April 2011

Mucins are high molecular weight glycoproteins with a variety of postulated biological functions, including physicochemical protection from toxins and mutagens, adhesion modulation, signal transduction, and regulation of cell growth. Mucins are widely and differentially expressed in the gastrointestinal tract. To date, studies of cellular expression have relied on light microscopy using in situ hybridization and immunohistochemistry. Although informative, it has been difficult with these techniques to ascertain exactly which cell types are producing a given mucin. We studied expression of MUC1, MUC2, and MUC4 apomucins in a series of normal colon biopsies using a combination of immunoelectron microscopy and light microscopy. MUC1 mucin was localized to both goblet and columnar cells, where it was seen in secretory vesicles, microvilli, and in cytoplasmic remnants in goblet cell thecae. MUC2 expression was restricted to goblet cells, in which reactivity was concentrated in the rough endoplasmic reticulum (RER). MUC4 expression was seen in both columnar and goblet cells, localized to the RER. The inability to detect MUC2 and MUC4 apomucins in the Golgi complex and the mature mucous gel probably represents masking of peptide epitopes following O-glycosylation. This study has helped clarify lineage-specific mucin synthesis in the normal colon. (J Histochem Cytochem 47:1063–1074, 1999)
Nuclear β-catenin in Colorectal Tumors: To Freeze or Not To Freeze?
Authors: Muriné Assumpta , Fabre Myriam , Mariñoso M. Luisa , Gallén Manel , Real Francisco X.
Published: April 2011

β-Catenin mediates the interaction of E-cadherin with α-catenin and the actin cytoskeleton. Recent evidence indicates that when the tumor suppressor gene APC is inactivated, β-catenin can translocate to the nucleus, where it acts as a transcriptional regulator. Because APC is inactivated in most colorectal cancers, β-catenin nuclear localization would be expected in these tumors. In a study of adhesion molecule expression in frozen colorectal cancer tissues, we were surprised by failure to detect nuclear β-catenin. Here we compared the reactivity of an anti-β-catenin monoclonal antibody with 11 colorectal cancers using immunohistochemistry on sections of frozen or paraffin-embedded samples. β-Catenin was never detected in the nuclei of normal or tumor cells in frozen tissue sections. By contrast, in 8/11 cases it was detected in the nuclei of tumor cells but not of normal cells in paraffin-embedded tissue sections. These results were confirmed with an independent rabbit polyclonal anti-β-catenin serum. We also examined β-catenin distribution in SW480 colon cancer cells, in which its nuclear accumulation has been reported. As in tissues, nuclear β-catenin was detected in paraffin-embedded but not in frozen samples. These findings are relevant because of the increasing interest in the study of β-catenin in tumors, based on its dual role in cell adhesion and transcriptional regulation. (J Histochem Cytochem 47:1089–1094, 1999)
Degradation of Overexpressed Wild-type and Mutant Uricase Proteins in Cultured Cells
Authors: Yokota Sadaki , Kamijo Keiju , Oda Toshiaki
Published: April 2011

Wild-type and mutated urate oxidase (UO) proteins were overexpressed in Cos-1 and HEK293 cells and were analyzed by Western blotting and several morphological methods. By immunoelectron microscopy, wild-type UO formed large aggregates in the cytoplasm and nucleoplasm and exhibited a crystalloid structure. Mutated UO (UOdC), from which 28 amino acids, including peroxisomal targeting signal at the C-terminus, were deleted, formed dispersed aggregates in the cytoplasm and nucleus. Chimeric UO (MUOdC), which was made by addition of the mitochondrial targeting signal of serine:pyruvate/glyoxylate aminotransferase to the N-terminus of UOdC, attached to ER to form a complicated MUOdC-ER complex. These three structures were immunostained for ubiquitin- and p32-subunits of proteasomes. Western blotting showed strong signal for UO and UOdC but very weak signal for MUOdC. The results suggest that overexpressed UO and UOdC accumulate in the cells because their synthesis rate is higher than the degradation rate, whereas MUOdC forming a complex with ER is degraded very rapidly. The ubiquitin-proteasome pathway may be involved in the degradation of these proteins.
Localization of MCC (Mutated in Colorectal Cancer) in Various Tissues of Mice and Its Involvement in Cell Differentiation
Authors: Senda Takao , Matsumine Akihiko , Yanai Hiroyuki , Akiyama Tetsu
Published: April 2011

Localization of the MCC (mutated in colorectal cancer) gene product, a cell cycle-regulating protein mutated in several colorectal tumors, in various mouse tissues was examined by immunohistochemistry and immunoelectron microscopy. MCC was localized on microvilli and in the apical cytoplasm in renal proximal tubule epithelial cells and pancreatic acinar cells. In hepatocytes, MCC was exclusively detected on microvilli. MCC was highly expressed in the cerebral cortex and the molecular layer of the cerebellar cortex and was partially associated with membrane organelles in neuronal elements. Adrenal chromaffin cells showed little expression of MCC. MCC was localized to the cell margins of ependymal cells, thyroid follicular cells, and anterior pituitary cells. In parotid acinar cells, only the apical surface was immunopositive. MCC was not expressed in skeletal and cardiac muscle. MCC was present at lateral cell borders in the duodenum and colon epithelium. In addition, the apical cytoplasm of colon epithelial cells exhibited intense immunoreactivity. The amount of MCC increased during differentiation of NGF-treated PC12 cells. In conclusion, MCC was expressed in differentiated cells and was associated with the plasma membrane and membrane organelles. In addition to the negative regulation of the cell cycle, MCC may be involved in cell differentiation.
Expression of Adrenomedullin and Proadrenomedullin N-terminal 20 Peptide in Human and Rat Prostate
Authors: Jiménez Nuria , Calvo Alfonso , Martínez Alfredo , Rosell David , Cuttitta Frank , Montuenga Luis M.
Published: April 2011

Adrenomedullin (AM) and proadrenomedullin N-terminal 20 peptide (PAMP) are two recently discovered hypotensive peptides translated from the same message transcript (preproAM mRNA). In this article we report the presence of AM, PAMP, and their mRNA in human and rat prostate and of AM receptor mRNA in rat prostate. PreproAM mRNA was found in the epithelium of normal human and rat prostate glands by in situ hybridization. In humans, it was mainly expressed in the basal cells. In rat, its expression was higher in the ducts than in the acini of all the prostate lobes. Immunocytochemistry identified a similar distribution pattern for AM compared with its mRNA but showed different locations for AM and PAMP immunoreactivity. The former was widespread in the epithelia, whereas the latter was almost exclusively found in neuroendocrine cells. In rat, Western blot analysis confirmed the presence of high levels of AM peptide in the ventral lobe and of its precursor in the ventral and dorsolateral lobes. Immunoreactivity for serotonin, chromogranin A, PAMP, and AM defined four subpopulations of prostate neuroendocrine-like cells in rat, a cell type that has not been previously described.
Selective but Nonspecific Immunolabeling of Enamel Protein-associated Compartments by a Monoclonal Antibody Against Vimentin
Authors: Josephsen Kaj , Smith Charles E. , Nanci Antonio
Published: April 2011

Vimentin, an intermediate filament component, has been identified in many mesenchymal cells by a variety of LM and EM immunolabeling techniques. In our study, several tissue-processing conditions and monoclonal and polyclonal antibodies against vimentin were screened for immunostaining of rat incisor odontoblasts. Using postembedding colloidal gold immunocytochemistry, we were unable to detect any convincing vimentin antigenicity in these cells, but one of the monoclonal antibodies (V9-S) unexpectedly resulted in intense labeling over intra- and extracellular compartments that normally are strongly immunoreactive with anti-amelogenin antibodies. Blocking experiments showed that V9-S binding was competed by anti-amelogenin antibody. Immunoblots indicated that enamel proteins reacted with this anti-vimentin antibody after fixation with glutaraldehyde. These data suggest that the observed immunoreaction is directed against an epitope apparently created by crosslinking of enamel proteins during fixation. Although the labeling cannot be considered specific, it is nevertheless selective because it is very precisely localized over compartments containing enamel proteins and shows no binding to other calcified dental tissues, including dentin and bone. The V9-S antibody can therefore be used as a reliable probe to identify the presence and distribution of amelogenins in fixed tissues.
Stage-dependent Redistribution of the V-ATPase During Bovine Implantation
Authors: Skinner Mhairi A. , MacLaren Leslie A. , Wildeman Alan G.
Published: April 2011

The 16-kD subunit of the vacuolar H+ -ATPase (V-ATPase), or ductin, is essential for the activity of this proton pump and has roles in intercellular communication and control of cell growth and differentiation. The V-ATPase is important for acidification-dependent degradation of tissue matrices through which some cell types move, and for pH regulation across some epithelial cell layers. Placentation involves intricate signaling, cell proliferation, and controlled invasion. We examined the distribution of three subunits of the V-ATPase in bovine trophoblast and endometrium at the time of implantation to determine the relationship of ductin expression to that of two other subunits, A (approximately 73 kD) and B (approximately 58 kD). Epithelial expression of all three subunits was observed, and in nonpregnant animals this expression was apical. As pregnancy proceeded, expression of all subunits became pericellular in luminal but not glandular epithelium, suggesting a redistribution of V-ATPase activity. The trophoblast expressed all three subunits during initial contact with the epithelium. In the stroma, ductin expression was reduced after implantation, and we discuss the possibility that ductin plays a role in the shifting communication between stromal and epithelial cells induced by embryo attachment.
Microtubule Nucleating Capacity of Centrosomes in Tissue Sections
Authors: Salisbury Jeffrey L. , Lingle Wilma L. , White Richard A. , Cordes Lynn E.M. , Barrett Susan
Published: April 2011

We used a novel adaptation of methods for microtubule polymerization in vitro to assess the MTOC activity of centrosomes in frozen-sectioned tissues. Remarkably, centrosomes of tissue sections retain the ability to nucleate microtubules even after several years of storage as frozen tissue blocks. Adaptations of these methods allow accurate counts of microtubules from individual cells and the quantitative estimation the MTOC activity of the intact tissue. These methods can be utilized to characterize MTOC activity in normal and diseased tissues and in particular tissues at different stages of development.
Localization of the Mixed-lineage Kinase DLK/MUK/ZPK to the Golgi Apparatus in NIH 3T3 Cells
Authors: Douziech Mélanie , Laberge Gino , Grondin Gilles , Daigle Nathalie , Blouin Richard
Published: April 2011

DLK/MUK/ZPK is a serine/threonine kinase that belongs to the mixed-lineage (MLK) subfamily of protein kinases. As is the case for most members of this family, relatively little is known about the physiological role of DLK/MUK/ZPK in mammalian cells. Because analysis of subcellular distribution may provide important clues concerning the potential in vivo function of a protein, an antiserum was generated against the amino terminal region of murine DLK/MUK/ZPK and used for localization studies in wild-type NIH 3T3 cells. Light microscopic immunocytochemistry experiments performed with the antiserum revealed that DLK/MUK/ZPK was specifically localized in a juxtanuclear structure characteristic of the Golgi complex. In support of this, treatment of cells with brefeldin A, a drug known to disintegrate the Golgi apparatus, caused disruption of DLK/MUK/ZPK peri-nuclear staining. Ultrastructural observation of NIH 3T3 cells also confirmed this localization, showing that most of the immunoreactivity was detected on membranes of the stacked Golgi cisternae. Consistent with localization studies, biochemical analyses revealed that DLK/MUK/ZPK was predominantly associated with Golgi membranes on fractionation of cellular extracts and was entirely partitioned into the aqueous phase when membranes were subjected to Triton X-114 extraction. On the basis of these findings, we suggest that DLK/MUK/ZPK is a peripheral membrane protein tightly associated with the cytoplasmic face of the Golgi apparatus.
Analysis of Stained Objects in Histological Sections by Spectral Imaging and Differential Absorption
Authors: Ornberg Richard L. , Woerner B. Mark , Edwards Dorothy A.
Published: April 2011

We describe a new light microscopic imaging system and method to perform high through put color image analysis on histological tissue sections. The system features a computer-controlled, random-access liquid crystal tunable filter and high-resolution digital camera on a conventional brightfield microscope. For any combination of stains, the method determines the spectral transmittance of each stain on the slide and selects two or more wavelengths at which the differential absorption between stain and counterstain is greatest and the exposure time is reasonably short. Flatfield corrected digital images at these wavelengths are acquired and divided to produce a gray scale ratio image. The ratio image is calculated such that the stained features of interest are highlighted above a uniform background and the counterstained features are highlighted below background. Image threshold procedures using either visual inspection or a threshold value determined by the image mean intensity and standard deviation are used to segment the stained features of interest for subsequent morphometry. Results are presented for peroxidase-AEC-labeled tumor tissue and trichrome-stained biomaterial implant tissues. In principle, the method should work for any combination of colored stains.
High Expression of MDM2 Protein and Low Rate of p21WAF1/CIP1 Expression in SCID Mice Epstein Barr Virus-induced Lymphoproliferation
Authors: Mansouri Saïd El , Martin Antoine , Mercadier Anne , Capoulade Corrine , Maréchal Vincent , Wiels Joëlle , Feuillard Jean , Raphaël Martine
Published: April 2011

To study the prevalence of p53 inactivation and MDM2/p21WAFI/CIP1 expression in severe combined immunodeficient (SCID) mice Epstein-Barr virus (EBV)-induced lymphoproliferation, 19 samples obtained after IP injection of peripheral blood mononuclear cells (PBMCs) from EBV-seropositive donors or lymphoblastoid cell lines (LCL) were analyzed. In all samples tested, overexpression of Ki-67 antigen was shown by immunohistochemistry, indicating a high proliferative index of SCID mice EBV-induced lymphoproliferation. P53 mutations were screened by functional assay in yeast in 14 samples. With this test, a p53-inactivating mutation was found in only one case; the remaining cases exhibited a wild-type p53 pattern. However, an accumulation of p53 protein was detected by immunohistochemistry in six of 19 samples. P21 expression was found in seven of 19 samples but was not correlated with the rate of p53 protein in tumors. In contrast, high levels of nuclear accumulation of MDM2 were found in all samples by immunohistochemistry. These results suggest that a high Ki-67 proliferative index in SCID mice EBV-induced lymphoproliferation is not due to the inactivation of p53 by mutation, but could be associated with an overexpression of MDM2, which would act by a p53-independent mechanism.
Immunohistochemical Evidence for the NO cGMP Signaling Pathway In Respiratory Ciliated Epithelia of Rat
Authors: Zhan Xinhua , Li Dechun , Johns Roger A.
Published: April 2011

Airway epithelia play a crucial role in protecting the lung from the external environment. Ciliated airway epithelial cells contribute to mucociliary transport systems via ciliary beating and electrolyte transport mechanisms to defend against respiratory tract infection. Both of these activities are regulated by nitric oxide (NO)-dependent mechanisms. To better understand the role of the NO-cGMP signal transduction cascade in these responses, we investigated the localization of endothelial nitric oxide synthase (eNOS), soluble guanylyl cyclase (sGC), cGMP-dependent protein kinase (PKG) I-α, and PKG I-β in the tracheas and lungs of normal rats by immunohistochemistry. Mouse anti-eNOS, rabbit anti-sGC, PKG I-α, and PKG I-β antibodies were used. Strong immunostaining for eNOS was detected in ciliated tracheal, bronchial, and bronchiolar epithelia, in Clara cells, and in Type II alveolar cells. The pattern of sGC and PKG I-β immunostaining showed striking parallels with that of eNOS staining. No staining was detectable in ciliated epithelium with the anti-PKG I-α antibody. Taken together, these observations suggest that PKG I-β might transduce NO-sGC signaling into biological responses in ciliated respiratory epithelia.
Differentiation of Separated Mouse Mammary Luminal Epithelial and Myoepithelial Cells Cultured on EHS Matrix Analyzed by Indirect Immunofluorescence of Cytoskeletal Antigens
Authors: Smalley Matthew J. , Titley Jenny , Paterson Hugh , Perusinghe Nina , Clarke Catherine , O'Hare Michael J.
Published: April 2011

We have previously demonstrated that purified virgin mouse mammary luminal epithelial and myoepithelial cells promiscuously express cell type-specific cytokeratins when they are cloned in vitro. Changes in cytokeratin expression may be indicators of the loss or change of the differentiated identity of a cell. To investigate the factors that may be responsible for the maintenance of differentiated cellular identity, specifically cell-cell and cell-matrix interactions, we cloned flow-sorted mouse mammary epithelial cells on the extracellular matrix (ECM) derived from the Engelbreth-Holm-Swarm murine sarcoma (EHS matrix). Changes in cell differentiation on EHS, compared with culture on glass, were analyzed by comparing patterns of cytokeratin expression. The results indicate that ECM is responsible for maintenance of the differentiated identity of basal/myoepithelial cells and prevents the inappropriate expression of luminal antigens seen on glass or plastic. Luminal cell identity in the form of retention of luminal markers and absence of basal/myoepithelial antigens, on the contrary, appears to depend on homotypic cell-cell contacts and interactions. The results also show that luminal cells (or a subpopulation of them) can generate a cell layer that expresses only basal cytokeratin markers (and no luminal cytokeratin markers) and may form a pluripotent compartment.
Differential Expression and Activity of Tissue-nonspecific Alkaline Phosphatase (TNAP) in Rat Odontogenic Cells In Vivo
Authors: Hotton Dominique , Mauro Nicole , Lézot Frédéric , Forest Nadine , Berdal Ariane
Published: April 2011

Among the four existing isoforms of alkaline phosphatase (AP), the present study is devoted to tissue-nonspecific alkaline phosphatase (TNAP) in mineralized dental tissues. Northern blot analysis and measurements of phosphohydrolase activity on micro-dissected epithelium and ectomesenchyme, in situ hybridization, and immunolabeling on incisors confirmed that the AP active in rodent teeth is TNAP. Whereas the developmental pattern of TNAP mRNA and protein and the previously described activity were similar in supra-ameloblastic and mesenchymal cells, they differed in enamel-secreting cells, the ameloblasts. As previously shown for other proteins involved in calcium and phosphate handling in ameloblasts, a biphasic pattern of steady-state TNAP mRNA levels was associated with additional variations in ameloblast TNAP protein levels during the cyclic modulation process. Although the association of TNAP upregulation and the initial phase of bio-mineralization appeared to be a basic feature of all mineralized tissues, ameloblasts (and to a lesser extent, odontoblasts) showed a second selectively prominent upregulation of TNAP mRNA/protein/activity during terminal growth of large enamel crystals only, i.e., the maturation stage. This differential expression/activity for TNAP in teeth vs bone may explain the striking dental phenotype vs bone reported in hypophosphatasia, a hereditary disorder related to TNAP mutation.
Expression of N-acetyllactosamine and β1,4-Galactosyltransferase (β4GalT-I) During Adenoma-Carcinoma Sequence in the Human Colorectum
Authors: Ichikawa Tetsuro , Nakayama Jun , Sakura Naoki , Hashimoto Tadashi , Fukuda Minoru , Fukuda Michiko N. , Taki Takao
Published: April 2011

We set out to determine the expression profiles of glycoproteins possessing N-acetyllactosamine, a precursor carbohydrate of sialyl Lex, during colorectal cancer development. We immunohistochemically analyzed the distribution of N-acetyllactosamine as well as of β4GalT-I, a member of the β1,4-galactosyltransferase family responsible for N-acetyllactosamine biosynthesis, in normal mucosa and in adenoma and carcinoma of the human colorectum. Using monoclonal antibody H11, N-acetyllactosamine was barely detectable in the normal mucosa. In low-grade adenoma, however, N-acetyllactosamine was weakly but definitely expressed on the cell surface, and its expression level was moderately increased in high-grade adenoma and markedly increased in carcinoma in situ as well as in advanced carcinoma. To detect β4GalT-I, we used a newly developed polyclonal antibody (designated A18G), which is specific for the stem region of human β4GalT-I. Faint expression of β4GalT-I was detectable in normal mucosa, and the expression level was moderately increased in low-grade adenoma and in high-grade adenoma and markedly increased in carcinoma in situ and advanced carcinoma. The expression of N-acetyllactosamine was highly correlated with the expression of β4GalT-I in these tumor cells. These results indicate that the expression level of β4GalT-I is apparently enhanced during tumorigenesis in the colorectum and that β4GalT-I mostly directs the carcinoma-associated expression of N-acetyllactosamine on the colorectal tumor cell surface.
Development and Aging of the Articular Cartilage of the Rabbit Knee Joint: Distribution of Biglycan, Decorin, and Matrilin-1
Authors: Kavanagh Emma , Ashhurst Doreen E.
Published: April 2011

We determined the distributions of the small proteoglycans biglycan and decorin and the glycoprotein matrilin-1 (cartilage matrix protein) during development and aging of articular cartilage in the rabbit knee joint. Before cavitation, the matrices of the interzone and the adjacent epiphyseal cartilage do not contain biglycan or decorin, but some chondrocytes express their mRNAs. Matrilin-1 is found only in the deeper epiphyseal cartilage. After cavitation, biglycan and decorin are detected in the presumptive articular cartilage, but there is no matrilin-1. All are present in the underlying epiphyseal cartilage. In the neonate, the epiphyseal cartilage is ossified and the articular cartilage becomes a discrete layer. Biglycan and decorin accumulate in the articular cartilage, but matrilin-1 remains confined to the residual epiphyseal cartilage. In the adult, the distributions of biglycan and decorin are highly variable. Decorin tends to be confined to the central region; matrilin-1 is absent. The findings indicate that the articular and epiphyseal cartilages are different from the earliest developmental stages. The epiphyseal cartilage can be identified by its possession of matrilin-1. Epiphyseal cartilage is removed during development to leave the articular cartilage. The relationships between the distributions of decorin and matrilin-1 and the fibrillar collagens are discussed.
100 Horsemen and the empty city: A game theoretic examination of deception in Chinese military legend
Authors: Cotton Christopher , Chang Liu
Published: April 2011

We present game theoretic models of two of the most famous military bluffs from history. These include the legend of Li Guang and his 100 horsemen (144 BC), and the legend of Zhuge Liang and the Empty City (228 AD). In both legends, the military commander faces a much stronger opposing army, but instead of ordering his men to retreat, he orders them to act in a manner consistent with baiting the enemy into an ambush. The stronger opposing army, uncertain whether it is facing a weak opponent or an ambush, then decides to flee and avoid battle. Military scholars refer to both stories to illustrate the importance of deception in warfare, often highlighting the creativity of the generals’ strategies. We model both situations as signaling games in which the opponent is uncertain whether the general is weak (i.e. has few soldiers) or strong (i.e. has a larger army waiting to ambush his opponent if they engage in combat). We then derive the unique Perfect Bayesian Equilibrium of the games. When the probability of a weak general is high enough, the equilibrium involves mixed strategies, with weak generals sometimes fleeing and sometimes bluffing about their strength. The equilibrium always involves the generals and their opponents acting as they did in the historical examples with at least a positive probability. When the probability of a weak general is lower (which is reasonable given the reputations of Li Guang and Zhuge Liang), then the unique equilibrium always involves bluffing by the general and retreat by his opponent.
“Making Space” for Ourselves
Authors: Venzant Chambers Terah T. , McCready Lance T.
Published: April 2011

Drawing from two separate case studies, one on lower track African American students and another on gay and gender nonconforming African American male students, this article explores how students with multiple stigmatized identities make sense of and respond to their marginalization, a process we term making space. In particular, we consider how making space can support students’ psychosocial needs and at the same time work against school engagement and academic striving. We describe types of “making space” strategies: sociospatial, performative, and political/institutional, and use these categories to describe the ways students in our projects responded to their perceived marginalization. Institutional processes that make these responses necessary are addressed as well as how schools can either mediate or intensify students’ feelings of marginalization and therefore their perceived need to “make space.”
‘One Size Fits All’? An Empirical Study into the Multidimensionality of Social Cohesion Indicators in Belgian Local Communities
Authors: Botterman Sarah , Hooghe Marc , Reeskens Tim
Published: April 2011

Recently, various attempts have been undertaken to develop a comprehensive social cohesion index to monitor the social progress of society. However, classical sociological theories predict that contemporary modern communities function according to a different type of social cohesion than traditional communities. From both a theoretical and a methodological perspective, it can be questioned whether social cohesion can be summarised into one single index. In this article, an extensive dataset of social cohesion indicators is analysed, including indicators of religious involvement, social inclusion, crime and voter turnout, for 308 local communities in the Flemish region of Belgium. Based on first- and second-order factor analyses, it is concluded that it is impossible to construct one single indicator for social cohesion when taking the multidimensionality of the concept into account. While the traditional form prevails in rural areas, it does not coincide with a distinct modern form that prevails in urban areas.
Multiple Transformations
Authors: Sýkora Luděk , Bouzarovski Stefan
Published: April 2011

This paper develops a conceptual framework for interpreting the process of urban change in post-communist cities. The departure from the legacies of the communist past has been effected through multiple transformation dynamics of institutional, social and urban change. While institutional reforms have been largely accomplished, the adjustment of urban land use patterns to new societal conditions is still ongoing. Hence, post-communist cities are still cities in transition. Using this interpretative framework and referring to a wide spectrum of academic work, the paper provides an overview of urban restructuring in post-communist countries over the past two decades with a specific focus on the examples of mutual integration of the three fields of transformation.
A New Institutionalism? The English School as International Sociological Theory
Authors: Schouenborg Laust
Published: April 2011

In this article I engage with the theoretical opening provided by Barry Buzan’s From International to World Society? I present an argument for five functional categories, which should be able to encompass all the institutions identified by English School scholars throughout history. Their introduction should point the way towards a sounder analytical framework for the study of what Buzan believes should be the new subject of the discipline of International Relations (IR). This subject is defined as second-order societies, meaning societies ‘where the members are not individual human beings, but durable collectivities of humans possessed of identities and actor qualities that are more than the sum of their parts’, and where the content of these societies, and the key object of analysis, is primary institutions. The purpose of the five functional categories is to break down this ‘social whole’ and provide a set of lenses through which to potentially analyse international societies throughout history.
Implementation of a Diabetes Self-management Education Program in Primary Care for Adults Using Shared Medical Appointments
Authors: Sanchez Iris
Published: April 2011

Purpose

The purpose of this study was to implement diabetes self-management education in primary care using the Chronic Care Model and shared medical appointments (SMA) to provide evidence-based interventions to improve process and measure outcomes.

Methods

A quality improvement project using the Plan-Do-Check-Act cycle was implemented in a primarycare setting in South Texas to provide diabetes self-management education for adults. Biological measures were evaluated in 70 patients at initiation of the project and thereafter based on current practice guidelines.

Results

The results of the project were consistent with the literature regarding the benefits, sustainability, and viability of SMA. As compared with that in studies presented in the literature, the patient population who participated in SMA had similar outcomes regarding improvement in A1C, self-management skills, and satisfaction. SMA are an innovative system redesign concept with the potential to provide comprehensive and coordinated care for patients with multiple and chronic health conditions while still being an efficient, effective, financially viable, and sustainable program.

Conclusions

As the incidence and prevalence of diabetes increase, innovative models of care can meet the growing demand for access and utilization of diabetes self-management education programs. Programs focusing on chronic conditions to improve outcomes can be replicated by health care providers in primary care settings. SMA can increase revenue and productivity, improve disease management, and increase provider and patient satisfaction.
Surveying Nurses
Authors: VanGeest Jonathan , Johnson Timothy P.
Published: April 2011

While surveys of nurses are an important tool in health services and policy research, they are often characterized by low response rates. The authors conducted a systematic review of 22 published reports of efforts to improve response rates to nurse surveys. Two general strategies were explored in this literature: incentive and design-based approaches. Even small financial incentives were found to be effective in improving nurse survey participation. Token nonmonetary incentives, in contrast, were much less effective. In terms of design strategies, postal and telephone strategies have generally been more successful than have fax or web-based approaches, with evidence also supporting use of mixed-mode surveys in this population. In addition, use of first-class stamps on return envelopes as well as questionnaires personalized and endorsed by legitimizing professional associations were also more likely to be successful. Researchers should continue to implement evidence-based strategies in order to improve the survey response of nurses.
Envoy Esteem Totally Implantable Hearing System
Authors: Kraus Eric M. , Shohet Jack A. , Catalano Peter J.
Published: April 2011

Objectives

(1) To assess outcomes of the Envoy Esteem Totally Implantable Hearing System as measured by hearing results compared with preimplant baseline unaided (BLU) and best‐fit aided conditions (BLA) and (2) to determine safety of the device.

Study Design

Prospective, nonrandomized, multicenter, subject‐as‐own‐control, US Food and Drug Administration (FDA) trial.

Setting

Private practice and hospital‐based.

Subjects and Methods

Between January 2008 and August 2009, an FDA trial was performed at 3 sites. Fifty‐seven subjects with bilateral, mild to severe sensorineural hearing loss, with discrimination greater than 40%, were implanted. Implanted components were (1) a sound processor and (2) 2 piezoelectric transducers (a sensor and a driver). A sound processor was implanted in the temporal bone. Transducers were coupled to the ossicles. Devices were activated 2 months postimplant. Hearing results were compared with ipsilateral BLU and BLA.

Results

Speech reception thresholds (SRTs) improved from BLA of 41.2 dB to 29.4 dB with the Esteem (P ≤. 001). Word recognition score (WRS) at 50 dB hearing level (HL) improved from BLA of 46.3% to 68.9% with the Esteem. Pure tone averages improved by 27 ± 1 dB (confidence interval, 30‐25). There were no changes in bone conduction. QuickSIN results showed no change. There were 6 serious adverse device effects: 2 wound infections (1 resolved medically, 1 required explantation), 1 delayed facial paralysis that resolved with medication, and 3 revisions due to limited benefit.

Conclusion

Phase 2 results at 12 months post implant demonstrated that (1) hearing results with the device are statistically superior to baseline best‐fit hearing aids for SRT and WRS and (2) the device is safe.
Topographic Gene Expression in the Sinonasal Cavity of Patients with Chronic Sinusitis with Polyps
Authors: Platt Michael P. , Soler Zachary M. , Kao Shyan‐Yuan , Metson Ralph , Stankovic Konstantina M.
Published: April 2011

Objective

To determine whether variations in gene expression exist at multiple subsites along the sinonasal tract in patients with chronic sinusitis with polyps and in healthy controls.

Study Design

Prospective, controlled study.

Setting

Academic medical center.

Subjects and Methods

Tissue expression levels of 5 genes, previously found to be characteristic of ethmoid polyps, were measured using real‐time quantitative polymerase chain reaction in 100 sinonasal tissue samples. Specimens harvested from 5 regions—the ethmoid sinus, septum, inferior turbinate, middle turbinate, and lateral nasal wall—in 10 patients with chronic sinusitis and ethmoid polyps were compared to tissue from similar regions in 10 control patients without sinusitis. Western blot analysis was performed to validate differential gene expression at the protein level.

Results

Gene expression levels of ethmoid polyps differed significantly from those of healthy ethmoid mucosa, as well as tissue from 4 surrounding anatomical sites in both patients with chronic sinusitis and controls. Alterations specific to the polyp tissue included downregulated genes, prolactin‐induced protein (fold change 377.2 ± 169.0, P <. 0001), and zinc α2‐glycoprotein (fold change 72.1 ± 26.5, P <. 0001), as well as upregulated genes, met proto‐oncogene (fold change 2.5 ± 0.7, P =. 029), and periostin (fold change 7.5 ± 3.4, P =. 003). No significant differences in gene expression was found for neurabin 2 (fold change 1.0, P =. 99).

Conclusion

The transcriptional pattern of ethmoid polyps appears to be unique compared with other subsites in the sinonasal cavity of patients with chronic sinusitis. Care must be taken when collecting specimens for molecular studies of the sinonasal tract to differentiate polyp from nonpolyp tissue in chronic sinusitis.