BACKGROUND AND OBJECTIVES: The aim of this study is to determine whether the hyperproliferative and hyperkeratotic characters of cholesteatoma are associated with differentiation of keratinocytes in cholesteatoma by examining the localization of marker proteins, such as involucrin, filaggrin, and cytokeratins. MATERIALS AND METHODS: Immunohistochemical study was carried out in 30 cholesteatoma tissues and 10 retroauricular skins to examine the expression of involucrin, filaggrin, cytokeratin 4, 10 and 16. The staining results were graded as negative, weakly positive (<10%), moderately positive (10-70%), and strongly positive (>70%). RESULTS: Involucrin was strongly expressed in upper spinous, granular, and corneal layer of cholesteatoma. Filaggrin was strongly expressed in granular and corneal layer of cholesteatoma. Cytokeratin 4 was expressed in basal layer of retroauricular skin, but occasionally expressed in suprabasal layer of cholesteatoma. Cytokeratin 10 was homogenously expressed in all suprabasal layer of retroauricular skin, whereas pattern of shift to surface layer was showed in cholesteatoma. Cytokeratin 16 was moderately expressed at suprabasal layer in cholesteatoma. CONCLUSIONS: It can be suggested that early differentiation of suprabasal layer may lead to hyperdifferentiation and hyperkeratosis. Different expression of cytokeratins possibly indicates the altered differentiation of cholesteatoma.
Cholesteatoma ; Intermediate Filament Proteins ; Keratin-16 ; Keratin-4 ; Keratinocytes ; Keratins ; Protein Precursors ; Proteins ; Skin

BACKGROUND AND OBJECTIVES: The aim of this study is to determine whether the hyperproliferative and hyperkeratotic characters of cholesteatoma are associated with differentiation of keratinocytes in cholesteatoma by examining the localization of marker proteins, such as involucrin, filaggrin, and cytokeratins. MATERIALS AND METHODS: Immunohistochemical study was carried out in 30 cholesteatoma tissues and 10 retroauricular skins to examine the expression of involucrin, filaggrin, cytokeratin 4, 10 and 16. The staining results were graded as negative, weakly positive (<10%), moderately positive (10-70%), and strongly positive (>70%). RESULTS: Involucrin was strongly expressed in upper spinous, granular, and corneal layer of cholesteatoma. Filaggrin was strongly expressed in granular and corneal layer of cholesteatoma. Cytokeratin 4 was expressed in basal layer of retroauricular skin, but occasionally expressed in suprabasal layer of cholesteatoma. Cytokeratin 10 was homogenously expressed in all suprabasal layer of retroauricular skin, whereas pattern of shift to surface layer was showed in cholesteatoma. Cytokeratin 16 was moderately expressed at suprabasal layer in cholesteatoma. CONCLUSIONS: It can be suggested that early differentiation of suprabasal layer may lead to hyperdifferentiation and hyperkeratosis. Different expression of cytokeratins possibly indicates the altered differentiation of cholesteatoma.
Cholesteatoma ; Intermediate Filament Proteins ; Keratin-16 ; Keratin-4 ; Keratinocytes ; Keratins ; Protein Precursors ; Proteins ; Skin

Periodontal ligament(PDL) cells have been known as playing an important roles in periodontal regeneration and gingival fibroblasts are also important to periodontal regeneration by forming connective tissue attachment. There were rare studies about the gene expression patterns of PDL cells and gingival fibroblasts, therefore in this study, we tried cDNA microarray-based gene expression monitoring to explain the functional differences of PDL cells and gingival fibroblasts in vivo and to confirm the characteristics of PDL cells. Total RNA were extracted from PDL cells and gingival fibroblasts of same person and same passages, and mRNA were isolated from the total RNA using Oligotex mRNA midi kit(Qiagen) and then fluorescent cDNA probe were prepared. And microarray hybridization were performed. The gene expression patterns of PDL cells and gingival fibroblasts were quite different. About 400 genes were expressed more highly in the PDL cells than gingival fibroblasts and about 300 genes were more highly expressed in the gingival fibroblasts than PDL cells. Compared growth factor- and growth factor receptor-related gene expression patterns of PDL cells with gingival fibroblasts, IGF-2, IGF-2 associated protein, nerve growth factor, placental bone morphogenic protein, neuron-specific growth- associated protein, FGF receptor, EGF receptor-related gene and PDGF receptor were more highly expressed in the PDL cells than gingival fibroblasts. The results of collagen gene expression patterns showed that collagen type I, type III, type VI and type VII were more highly expressed in the PDL cells than gingival fibroblasts, and in the gingival fibroblasts collagen type V, XII were more highly expressed than PDL cells. The results of osteoblast-related gene expression patterns showed that osteoblast specific cysteine-rich protein were more highly expressed in the PDL cells than gingival fibroblasts. The results of cytoskeletal proteins gene expression patterns showed that alpha-smooth muscle actin, actin binding protein, smooth muscle myosin heavy chain homolog and myosin light chain were more highly expressed in the PDL cells than gingival fibrobalsts, and beta-actin, actin-capping protein(beta subunit), actin- related protein Arp3(ARP) and myosin class I(myh-1c) were more highly expressed in the gingival fibroblasts than PDL cells. Osteoprotegerin/osteoclastogenesis inhibitory factor(OPG/OCIF) was more highly expressed in the PDL cells than gingival fibroblasts. According to the results of this study, PDL cells and gingival fibroblasts were quite different gene expression patterns though they are the fibroblast which have similar shape. Therefore PDL cells & gingival fibroblasts are heterogeneous populations which represent distinct characteristics. If more studies about genes that were differently expressed in each PDL cells & gingival fibroblasts would be performed in the future, it would be expected that the characteristics of PDL cells would be more clear.
Actins ; Carrier Proteins ; Collagen ; Collagen Type I ; Collagen Type V ; Connective Tissue ; Cytoskeletal Proteins ; DNA, Complementary* ; Epidermal Growth Factor ; Fibroblasts* ; Gene Expression Profiling ; Gene Expression* ; Humans ; Insulin-Like Growth Factor II ; Muscle, Smooth ; Myosin Heavy Chains ; Myosin Light Chains ; Myosins ; Nerve Growth Factor ; Oligonucleotide Array Sequence Analysis* ; Osteoblasts ; Periodontal Ligament* ; Receptors, Fibroblast Growth Factor ; Receptors, Platelet-Derived Growth Factor ; Regeneration ; RNA ; RNA, Messenger

OBJECTIVETo study the clinicopathologic features of papillary tumor of the pineal region (PTPR).

METHODThree hundred and eighty six cases of pineal region and posterior third ventricle tumors, two newborn and two adult pineal glands were analyzed by HE, PAS and immunohistochemistry of 16 antibodies (EnVision method).

RESULTSFive cases of PTPR were diagnosed with mixed papillary features and densely cellular areas, and included one recurrent case. In the papillary areas, the vessels were lined by one or several layers of cuboidal/columnar cells; the vessel wall was hyalinized. In the densely cellular areas, sheets or nests of tumor cells were seen. The tumor cells of these five cases were immunoreactive to CK, CK8/18, synaptophysin, MAP2, nestin, S-100, and vimentin. Four cases were immunoreactive to NSE and CgA; and 2 cases were immunoreactive to NF. All five cases were negative for EMA, CK5/6, CEA, and NeuN. Ki-67 labeling index ranged from 1% to 6%.Three patients were alive, and the recurrent one died.

CONCLUSIONSPTPR occurs in patients with over a wide age range, from children to adults, and is more commonly found in male than female. PTPR is composed of both papillary and solid areas, characterized by epithelial cytology, and needs to be differentiated from ependymoma. PTPR may originate from the specialized ependymocytes of the subcommissural organ. The prognostic factors are early diagnosis, complete surgical resection and radiotherapy.

Adolescent ; Adult ; Brain Neoplasms ; diagnostic imaging ; metabolism ; pathology ; radiotherapy ; surgery ; Carcinoma, Papillary ; diagnostic imaging ; metabolism ; pathology ; radiotherapy ; surgery ; Child ; Diagnosis, Differential ; Ependymoma ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Keratin-18 ; metabolism ; Keratin-8 ; metabolism ; Keratins ; metabolism ; Male ; Microtubule-Associated Proteins ; metabolism ; Nestin ; metabolism ; Pineal Gland ; Pinealoma ; metabolism ; pathology ; S100 Proteins ; metabolism ; Synaptophysin ; metabolism ; Tomography, X-Ray Computed ; Vimentin ; metabolism ; Young Adult

To evaluate the clinical significance of autoantibodies to three major epithelial cytokeratins (CK) -- CK8, CK18, and CK19 -- we compared 66 patients with toluene diisocyanate (TDI)-induced asthma (group I) with three control groups: 169 asymptomatic exposed subjects (group II), 64 patients with allergic asthma (group III), and 123 unexposed healthy subjects (group IV). Serum IgG, specific for human recombinant CKs, were measured by ELISA (enzyme linked immunosorbent assay), and ELISA inhibition tests were performed. The existence of these antibodies was confirmed by IgG immunoblot analysis. Anti-TDI-HSA (human serum albumin) IgE and IgG antibodies were measured by ELISA in the same set of the patients. The prevalence of CK8, CK18, and CK19 auotantibodies in group I was significantly higher than in the other three groups. Results of the ELISA inhibition test showed significant inhibition with the addition of three CKs in a dose-dependent manner. No significant association was found between CK autoantibodies and the prevalence of anti- TDI-HSA IgG and IgE antibodies. These results suggest that autoantibodies to CK18 and CK19 can be used as serologic markers for identifying patients with TDI-induced asthma among exposed workers.
Toluene 2,4-Diisocyanate/*toxicity ; Sensitivity and Specificity ; Occupational Diseases/chemically induced/*diagnosis ; Middle Aged ; Male ; Keratins/*immunology ; Keratin-8/immunology ; Keratin-19/immunology ; Keratin-18/immunology ; Immunoblotting ; Humans ; Female ; Enzyme-Linked Immunosorbent Assay ; Biological Markers/blood ; Autoantibodies/*blood ; Asthma/chemically induced/*diagnosis ; Adult

OBJECTIVETo investigate the role of microtubule-associated protein 2 (MAP-2) and nestin in gastric development in human embryos and fetuses.

METHODSImmunohistochemistry was used to detect the expressions of MAP-2 and nestin proteins in the gastric cardia, pyloric and gastric tissues of human embryos and fetuses during the second, third and fourth month of development.

RESULTSIn the second to fourth months of gestation, MAP-2 and nestin expressions were detected in the neural cells and neural fibers of the intermuscular nerve plexus and submucosal plexus in the gastric cardia, pyloric and gastric tissues. As the gestational age increased, the number of MAP-2- and nestin-positive cells and the expression intensity all increased in the myenteric plexus, but MAP-2 and nestin expressions were negative in the glandular and mucosal tissues of human embryonic and fetal gastric cardia, pylorus or gastric walls.

CONCLUSIONMAP-2 and nestin participate in the regulation of the development of gastric tissues in human embryos.

Fetus ; metabolism ; Humans ; Intermediate Filament Proteins ; metabolism ; Microtubule-Associated Proteins ; metabolism ; Nerve Tissue Proteins ; metabolism ; Nestin ; Stomach ; embryology ; metabolism

Carcinoma, Renal Cell ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Humans ; Keratin-18 ; metabolism ; Keratin-19 ; metabolism ; Keratin-8 ; metabolism ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Nephrectomy ; Neprilysin ; metabolism ; Racemases and Epimerases ; metabolism

To study the cellular changes and the potential role of CD2-associated protein (CD2AP) in podocyte differentiation, conditionally immortalized murine podocyte cell line was cultured in RPMI 1640 medium under permissive condition at 33 °C. After transfection with CD2AP small interfering RNA (siRNA) the cells were shifted to non-permissive condition at 37 °C. Simultaneously, untransfected cells were taken as differentiation control. The podocyte proliferation rate was determined by MTT method. The expressions of CD2AP, WT1, synaptopodin and nephrin mRNAs were examined by RT-PCR. CD2AP, WT1 and nephrin protein expressions were examined by Western blot. The distribution of CD2AP, nephrin, F-actin and tubulin in differentiated and undifferentiated podocytes was detected by laser scanning confocal microscopy. The results showed: (1) CD2AP, WT1 and nephrin were stably expressed in differentiated and undifferentiated podocytes while synaptopodin was only expressed in differentiated podocytes. (2) CD2AP and nephrin mRNA and protein expressions were up-regulated during podocyte differentiation (P<0.05). (3) CD2AP and tubulin were distributed in the cytoplasm and perinulcear region in undifferentiated podocytes, and F-actin was predominantly localized to a cortical belt and paralleled to the cell axis. Under differentiation condition, CD2AP distribution profile was presented as peripheral accumulation, tubulin took on fascicular style and F-actin extended into foot processes in podocytes. CD2AP colocalized with nephrin and F-actin in undifferentiated podocytes. (4) After transfection with CD2AP siRNA, the expression of CD2AP was partially inhibited and cell growth was arrested; Synaptopodin, the differentiation podocyte marker, was apparently down-regulated; The differentiation of podocytes was delayed. The results demonstrate that podocyte differentiation is accompanied by cytoskeleton rearrangement and cell morphology change. CD2AP might play an essential role in podocyte differentiation.
Actins ; metabolism ; Adaptor Proteins, Signal Transducing ; physiology ; Animals ; Cell Differentiation ; Cell Line ; Cytoskeletal Proteins ; physiology ; Cytoskeleton ; metabolism ; Membrane Proteins ; metabolism ; Mice ; Microfilament Proteins ; metabolism ; Podocytes ; cytology ; RNA, Small Interfering ; Transfection ; WT1 Proteins ; metabolism

17, cytokeratin 18 antibody staining were used. There was significant difference of positive reaction between non-recurred ameloblastoma and recurred ameloblastoma in PCNA and cytokeratin 17. There were no significant difference of positive reaction between non-recurred ameloblastoma and recurred ameloblastoma in p53, Ki-67 and cytokeratin 18. From the above results, it is suggested that the recurrence of ameloblastoma is related to positive reactions of PCNA and cytokeratin 17 and the progonsis of the recurrence of ameloblastoma is able to be predicted by using PCNA and cytokeratin 17.]]>
Ameloblastoma ; Jaw ; Keratin-17 ; Keratin-18 ; Keratins ; Proliferating Cell Nuclear Antigen ; Recurrence

PURPOSE: the most important biological behavior of breast cancer is its invasive potential and many efforts was made to reveal the factors related with the invasiveness of breast cancer cells. Some researchers reported that intermediate filament biology could represent an emerging and exciting field in tumor biology with respect to tumor aggressiveness and invasiveness. There are some experimental evidences that co-expression of vimentin, a interfilament marker indicative of mesenchymal lineage, and cytokeratin interfilaments can be correlated with invasiveness and metastatic deposits. So, we tried to determine the role of intermediate filaments such as cytokeratins and vimentin with respect of bone marrow micrometastases. METHODS: Expression of cytokeratins 8, 18, 19 and viementin were immunohistochemically evaluated. Detection of bone marrow micrometastases was preformed through RT-PCR targeting mRNA of cytokeratin 19. In order to compare the study group by the expression extent of cytokeratins, the case expressing 50% or more of observed cells was classified into the case with high expression and the case expressing 49% or less was classified into the case with low expression. RESULTS: The only cytokeratin of high expression representing the risk of bone marrow micrometastases was cytokeratin 8. Vimentin expression by itself did not show any significance indicating bone marrow micrometastases. However, The cases possesing cytokeratin 8, 18, and 19 expression, altogether 75% or more showed a significantly high risk to bone marrow micrometastases. In that cases, addition of vimentin expression allowed a more higher possibility of bone marrow micrometastases. CONCLUSION: A high expression of cytokeratin 8 among cytokeratins was related with bone marrow metastases. However, vimentin expression by itself did not show any realtionship with bone marrow metastases. So, a further study is needed in order to reveal the role of vimentin expression in progression and metastases of breast cancer.
Biology ; Bone Marrow ; Breast Neoplasms* ; Breast* ; Intermediate Filaments ; Keratin-19 ; Keratin-8 ; Keratins ; Neoplasm Metastasis* ; Neoplasm Micrometastasis ; RNA, Messenger ; Vimentin*