Immunohistochemical localization of periostin in human gingiva

<p>The periostin is a matricellular protein expressed in collagen-rich tissues including some dental and periodontal tissues where it is regulated by mechanical forces, growth factors and cytokines. Interestingly the expression of this protein has been found modified in different gingival pathologies although the expression of periostin in normal human gingiva was never investigated. Here we used Western blot and double immunofluorescence coupled to laser-confocal microscopy to investigated the occurrence and distribution of periostin in different segments of the human gingival in healthy subjects. By Western blot a protein band with an estimated molecular mass of 94 kDa was observed. Periostin was localized at the epithelial-connective tissue junction, or among the fibers of the periodontal ligament, and never co-localized with cytokeratin or vimentin thus suggesting it is an extracellular protein. These results demonstrate the occurrence of periostin in adult human gingiva; its localization suggests a role in the bidirectional interactions between the connective tissue and the epithelial cells, and therefore in the physiopathological conditions in which these interactions are altered. </p>

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HGG-26. H3G34V MUTATION AFFECTS GENOMIC H3K36 METHYLATION IN PEDIATRIC GLIOMA

Neuro-Oncology ◽

10.1093/neuonc/noaa222.310 ◽

2020 ◽

Vol 22 (Supplement_3) ◽

pp. iii348-iii348

Author(s):

Tina Huang ◽

Andrea Piunti ◽

Elizabeth Bartom ◽

Jin Qi ◽

Rintaro Hashizume ◽

...

Keyword(s):

Western Blot ◽

Allelic Frequency ◽

Glioma Cell Line ◽

Fluorescent Imaging ◽

Wild Type ◽

Gene Expressions ◽

Pediatric Glioma ◽

Genes Encoding ◽

Normal Human

Abstract BACKGROUND Histone H3.3 mutation (H3F3A) occurs in 50% of cortical pediatric high-grade gliomas. This mutation replaces glycine 34 with arginine or valine (G34R/V), impairing SETD2 activity (H3K36-specific trimethyltransferase), resulting in reduced H3K36me on H3G34V nucleosomes relative to wild-type. This contributes to genomic instability and drives distinct gene expressions associated with tumorigenesis. However, it is not known if this differential H3K36me3 enrichment is due to H3G34V mutant protein alone. Therefore, we set to elucidate the effect of H3G34V on genomic H3K36me3 enrichment in vitro. METHODS Doxycycline-inducible short hairpin RNA (shRNA) against H3F3A was delivered via lentivirus to established H3G34V mutant pediatric glioma cell line KNS42, and H3G34V introduced into H3.3 wild type normal human astrocytes (NHA). Transfections were confirmed by western blot, fluorescent imaging, and flow cytometry, with resulting H3.3WT and H3K36me3 expression determined by western blot. H3.3WT, H3K36me3, and H3G34V ChIP-Seq was performed to evaluate genomic enrichment. RESULTS Complete knockdown of H3G34V was achieved with DOX-induced shRNA, with no change in total H3.3, suggesting disproportionate allelic frequency of genes encoding H3.3 (H3F3A and H3F3B). Modest increase in H3K36me3 occurred after H3F3A-knockdown from KNS42, suggesting H3G34V alone impacts observed H3K36me3 levels. Distinct H3K36me3 genomic enrichment was observed with H3G34V knock-in. CONCLUSIONS We demonstrate that DOX-inducible knockdown of H3F3A in an H3G34V mutant pediatric glioma cells and H3G34V mutation transduction in wild-type astrocytes affects H3K36me3 expression. Further evaluation by ChIP-Seq analysis for restoration of wild-type genomic H3K36me3 enrichment patterns with H3G34V knockdown, and mutant H3K36me3 patterns with H3G34V transduction, is currently underway.

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Immunohistochemical localization of extracellular matrix in various types of fibrinoid of the normal human term placenta

Placenta ◽

10.1016/s0143-4004(05)80575-2 ◽

1993 ◽

Vol 14 (4) ◽

pp. A55 ◽

Cited By ~ 1

Author(s):

Azizbeck K. Nanaev ◽

Andrej P. Milovanov ◽

Serguej P. Domogatsky

Keyword(s):

Extracellular Matrix ◽

Immunohistochemical Localization ◽

Human Term Placenta ◽

Term Placenta ◽

Normal Human

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Targeted Deletion of the SPARC Gene Accelerates Disc Degeneration in the Aging Mouse

Journal of Histochemistry & Cytochemistry ◽

10.1369/jhc.5a6687.2005 ◽

2005 ◽

Vol 53 (9) ◽

pp. 1131-1138 ◽

Cited By ~ 47

Author(s):

Helen E. Gruber ◽

E. Helene Sage ◽

H. James Norton ◽

Sarah Funk ◽

Jane Ingram ◽

...

Keyword(s):

Extracellular Matrix ◽

Connective Tissue ◽

Matricellular Protein ◽

Secreted Protein ◽

Wild Type ◽

Variable Size ◽

Targeted Deletion ◽

Ultrastructural Studies ◽

Aging Mouse ◽

Lower Lumbar

SPARC (secreted protein, acidic, and rich in cysteine) is a matricellular protein that is present in the intervertebral disc; in man, levels of SPARC decrease with aging and degeneration. In this study, we asked whether targeted deletion of SPARC in the mouse influenced disc morphology. SPARC-null and wild-type (WT) mice were studied at 0.3–21 months of age. Radiologic examination of spines from 2-month-old SPARC-null mice revealed wedging, endplate calcification, and sclerosis, features absent in age-matched WT spines. Discs from 3-month-old SPARC-null mice had a greater number of annulus cells than those of WT animals (1884.6 ± 397.9 [mean ± SD] vs 1500.2 ± 188.2, p=0.031). By 19 months discs from SPARC-null mice contained fewer cells than WT counterparts (1383.6 ± 363.3 vs 1466.8 ± 148.0, p=0.033). Histology of midsagittal spines showed herniations of lower lumbar discs of SPARC-null mice ages 14–19 months; in contrast, no herniations were seen in WT age-matched animals. Ultrastructural studies showed uniform collagen fibril diameters in the WT annulus, whereas in SPARC-null disc fibrils were of variable size with irregular margins. Consistent with the connective tissue deficits observed in other tissues of SPARC-null mice, our findings support a fundamental role for SPARC in the production, assembly, or maintenance of the disc extracellular matrix.

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Immunohistochemical localization of acidic and basic fibroblast growth factors in normal human endometrium and endometriosis and the detection of their mRNA by polymerase chain reaction

Human Reproduction ◽

10.1093/oxfordjournals.humrep.a137856 ◽

1993 ◽

Vol 8 (1) ◽

pp. 11-16 ◽

Cited By ~ 70

Author(s):

R.A. Ferriani ◽

D.S. Charnock-Jones ◽

A. Prentice ◽

E.J. Thomas ◽

S.K. Smith

Keyword(s):

Polymerase Chain Reaction ◽

Growth Factors ◽

Fibroblast Growth Factors ◽

Immunohistochemical Localization ◽

Human Endometrium ◽

Fibroblast Growth ◽

Chain Reaction ◽

Normal Human ◽

Polymerase Chain

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Ultrastructural and morphometrical evaluations on normal human dermal connective tissue – the influence of age, sex and body region

British Journal of Dermatology ◽

10.1111/j.1365-2133.1996.tb07935.x ◽

1996 ◽

Vol 134 (6) ◽

pp. 1013-1022 ◽

Cited By ~ 25

Author(s):

D. QUAGLINO ◽

G. BERGAMINI ◽

F. BORALDI ◽

I. PASQUALI RONCHETTI

Keyword(s):

Connective Tissue ◽

Body Region ◽

Normal Human ◽

Dermal Connective Tissue

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A unique protein in normal human cerebrospinal fluid.

Clinical Chemistry ◽

10.1093/clinchem/29.10.1842 ◽

1983 ◽

Vol 29 (10) ◽

pp. 1842-1844 ◽

Cited By ~ 4

Author(s):

N M Papadopoulos ◽

P A LeWitt ◽

R P Newman ◽

M I Raphaelson ◽

T N Chase

Keyword(s):

Cerebrospinal Fluid ◽

Serum Proteins ◽

Normal Volunteer ◽

Protein Band ◽

Serum Samples ◽

Immunofixation Electrophoresis ◽

Normal Human ◽

Monospecific Antibodies ◽

Immunoprecipitation Reaction ◽

Individual Human

Abstract On analysis of cerebrospinal fluid (CSF) samples from normal volunteer donors by high-resolution zone electrophoresis on agarose gel, an electrophoretically homogeneous protein band consistently appeared in the gamma-globulin region. Application of immunofixation electrophoresis in attempts to identify the band with use of monospecific antibodies against individual human serum proteins and against heavy- and light-chain immunoglobulins as well as polyvalent antisera did not produce a positive immunoprecipitation reaction with the protein band. The serum samples from these subjects did not show similar bands. Therefore, we conclude that this protein band is a normally occurring protein that is unique to CSF.

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Expression Profile of the Matricellular Protein Osteopontin in Primary Open-Angle Glaucoma and the Normal Human Eye

Investigative Opthalmology & Visual Science ◽

10.1167/iovs.11-7409 ◽

2011 ◽

Vol 52 (9) ◽

pp. 6443 ◽

Cited By ~ 13

Author(s):

Uttio Roy Chowdhury ◽

Seung-Youn Jea ◽

Dong-Jin Oh ◽

Douglas J. Rhee ◽

Michael P. Fautsch

Keyword(s):

Expression Profile ◽

Primary Open Angle Glaucoma ◽

Open Angle Glaucoma ◽

Matricellular Protein ◽

Open Angle ◽

Human Eye ◽

Normal Human

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Multiple isoforms of the ryanodine receptor are expressed in rat pancreatic acinar cells

Biochemical Journal ◽

10.1042/bj3510265 ◽

2000 ◽

Vol 351 (1) ◽

pp. 265-271 ◽

Cited By ~ 22

Author(s):

Timothy J. FITZSIMMONS ◽

Ilya GUKOVSKY ◽

James A. McROBERTS ◽

Edward RODRIGUEZ ◽

F. Anthony LAI ◽

...

Keyword(s):

Western Blot ◽

Ryanodine Receptor ◽

Acinar Cell ◽

Acinar Cells ◽

Protein Band ◽

Pancreatic Acinar Cell ◽

Pancreatic Acinar Cells ◽

Rt Pcr ◽

Pancreatic Acini ◽

Cell Functions

Regulation of cytosolic Ca2+ is important for a variety of cell functions. The ryanodine receptor (RyR) is a Ca2+ channel that conducts Ca2+ from internal pools to the cytoplasm. To demonstrate the presence of the RyR in the pancreatic acinar cell, we performed reverse transcriptase (RT)-PCR, Western blot, immunocytochemistry and microscopic Ca2+-release measurements on these cells. RT-PCR showed the presence of mRNA for RyR isoforms 1, 2 and 3 in both rat pancreas and dispersed pancreatic acini. Furthermore, mRNA expression for RyR isoforms 1 and 2 was demonstrated by RT-PCR in individual pancreatic acinar cells selected under the microscope. Western-blot analysis of acinar cell immunoprecipitates, using antibodies against RyR1 and RyR2, showed a high-molecular-mass (> 250kDa) protein band that was much less intense when immunoprecipitated in the presence of RyR peptide. Functionally, permeablized acinar cells stimulated with the RyR activator, palmitoyl-CoA, released Ca2+ from both basolateral and apical regions. These data show that pancreatic acinar cells express multiple isoforms of the RyR and that there are functional receptors throughout the cell.

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A Novel CD44 v3 Isoform is Involved in Head and Neck Squamous Cell Carcinoma Progression

Otolaryngology ◽

10.1067/mhn.2001.114674 ◽

2001 ◽

Vol 124 (4) ◽

pp. 426-432 ◽

Cited By ~ 31

Author(s):

Elizabeth J. Franzmann ◽

Donald T. Weed ◽

Francisco J. Civantos ◽

W. Jarrard Goodwin ◽

Lilly Y.W. Bourguignon

Keyword(s):

Squamous Cell Carcinoma ◽

Western Blot ◽

Cell Carcinoma ◽

Head And Neck ◽

Southern Blot ◽

Squamous Cell ◽

Blot Analysis ◽

Western Blot Analysis ◽

Southern Blot Analysis ◽

Double Immunofluorescence

OBJECTIVES: CD44 comprises a family of isoforms involved in tumorigenesis. Here we investigate the role of CD44 isoforms in head and neck squamous cell carcinoma (HNSCC) progression. MATERIALS AND METHODS: HNSCC specimens underwent reverse transcriptase-polymerase chain reaction (RT-PCR) followed by Southern blot analysis. After surface biotinylation, FaDu (hypopharyngeal HNSCC) and CD44v3-transfected COS-7 cells were CD44 antibody-precipitated and compared by Western blot analysis. FaDu cells underwent double immunofluorescence staining and growth assays. RESULTS: Southern blot analysis suggested differential CD44v3 isoform expression in tumor and normal tissue. Cloning and sequencing revealed 2 novel CD44v isoforms. Western blot analysis suggested CD44v3 expression in COS-7 transfectants and FaDu. Double immunofluorescence staining revealed colocalization of CD44v3 and actin in FaDu projections. Anti-CD44v3 antibody decreased FaDu growth. CONCLUSION: HNSCC tissue and FaDu appear to express CD44v3 isoforms. These isoforms may promote tumorigenesis. CLINICAL SIGNIFICANCE: CD44v3 isoforms may be effective tumor markers and targets for HNSCC therapy.

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