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Sample records for activity immunohistochemical analysis

  1. The immunohistochemical analysis of the proliferative activity and the maturity degree of lymphatic vessels in oral squamous cell carcinomas.

    PubMed

    Ciurea, Raluca Niculina; Stepan, Alex Emilian; Simionescu, Cristiana Eugenia; Mărgăritescu, Claudiu; Pirici, Daniel; Tica, Oana Sorina; Ciurea, Marius Eugen; Stepan, Desdemona; Enache, Oana Andreea; Vîlcea, Alina Maria; Stoica, Loredana Elena; Pătraşcu, Virgil

    2015-01-01

    Oral cancers still represent a major health problem; regional lymph node metastases occur in 30-40% of head and neck squamous cell carcinomas and are associated with unfavorable prognosis and decreased survival. The study included 35 cases of oral squamous cell carcinomas (OSCC), which were analyzed by double reactions to determine the proliferative activity (anti-human D2-40/Ki67) and the maturity degree (anti-human D2-40/α-SMA) of lymphatic vessels, both intratumoral (IT) and in the advancing edge (AE), and in relation to clinicopathological prognostic parameters. The mean values of D2-40 lymphatic vessel density (LVD) were higher in AE then in IT level. Poorly differentiated carcinomas, T3/T4, presented the highest LVD values, both IT and in the AE. LVD was higher in advanced stages and metastasizing carcinomas. Ki67 was positive in all cases, Ki67 proliferation index (IP) indicated higher values in poorly differentiated carcinoma, T3/T4, metastasizing ones, both IT and in the AE. LVD and IP Ki67 showed a positive linear correlation. D2-40/Ki67-positive vessels were identified only at the AE or close to it. D2-40/Ki67 LVD had highest values in advanced stages carcinoma, with metastases. D2-40/α-SMA-positive vessels were identified only in the neighborhood of the tumor and LVD highest values were present in early-stage carcinomas and without metastases. A negative linear correlation between proliferation and maturity of the lymphatic vessels was found. The study indicated a strong association between lymphatic proliferative activity and lymph node metastases, suggesting the need for targeted antilymphangiogenic therapies in OSCC.

  2. Noncollagenous proteins in heterotopic ossification. Immunohistochemical analysis in 15 paraplegies.

    PubMed

    Bosse, A; Wuisman, P; Jones, D B; Schwarz, K

    1993-12-01

    We used immunohistochemical techniques to investigate the distribution pattern of osteonectin, osteocalcin, bone sialoprotein II and the small proteoglycans decorin and PG 100 during different stages of heterotopic ossification (HO) in pressure sores of paraplegic patients. All these noncollagenous proteins (NCPs) accumulated in fibroblasts and preosteoblasts, predominantly in the activity centers of early osteogenetic areas. Mature types of HO showed a more discrete expression pattern for this protein group, with weaker reactions in the narrow osteoblastic rims. Decorin was detected predominantly in the stroma of HO. Our results indicate that the NCPs are important components during the pathogenesis of HO and that fibroblasts may serve as osteoprogenitor cells.

  3. [Immunohistochemical description of proliferative activity and apoptosis of lung squamous cell carcinoma (literature review)].

    PubMed

    Филенко, Борис Н; Ройко, Наталия В; Степанчук, Алла П; Проскурня, Сергей А

    2016-01-01

    The analysis of the publications are describe immunohistochemical study of proliferative activity and apoptosis of lung squamous cell carcinoma. Established that the imbalance between proliferation and cell death is a key process in the development of tumors. However, the value of tumor markers in histogenesis and morfogenesis of tumors and forecast their occurrence is not studied enough. Despite the significant amount of scientific literature devoted to this issue, has not yet established a clear link expression of immunohistochemical markers of proliferation and apoptosis with the degree of differentiation of squamous cell lung cancer. Analysis of the literature shows that the morphology of this histogenetics type lung cancer at the cellular, subcellular structural and functional levels are controversial and require detailed investigation. PMID:27487551

  4. [Immunohistochemical description of proliferative activity and apoptosis of lung squamous cell carcinoma (literature review)].

    PubMed

    Филенко, Борис Н; Ройко, Наталия В; Степанчук, Алла П; Проскурня, Сергей А

    2016-01-01

    The analysis of the publications are describe immunohistochemical study of proliferative activity and apoptosis of lung squamous cell carcinoma. Established that the imbalance between proliferation and cell death is a key process in the development of tumors. However, the value of tumor markers in histogenesis and morfogenesis of tumors and forecast their occurrence is not studied enough. Despite the significant amount of scientific literature devoted to this issue, has not yet established a clear link expression of immunohistochemical markers of proliferation and apoptosis with the degree of differentiation of squamous cell lung cancer. Analysis of the literature shows that the morphology of this histogenetics type lung cancer at the cellular, subcellular structural and functional levels are controversial and require detailed investigation.

  5. Apoptosis Activation in Human Carious Dentin. An Immunohistochemical Study

    PubMed Central

    Loreto, C.; Psaila, A.; Musumeci, G.; Castorina, S.; Leonardi, R.

    2015-01-01

    The exact mechanisms and enzymes involved in caries progression are largely unclear. Apoptosis plays a key role in dentin remodelling related to damage repair; however, it is unclear whether apoptosis in decayed teeth is activated through the extrinsic or the intrinsic pathway. This ex vivo immunohistochemical study explored the localization of TRAIL, DR5, Bcl-2 and Bax, the main proteins involved in apoptosis, in teeth with advanced caries. To evaluate TRAIL, DR5, Bcl-2 and Bax immunoexpressions twelve permanent carious premolars were embedded in paraffin and processed for immunohistochemistry. The results showed that TRAIL and DR5 were overexpressed in dentin and in pulp vessels and mononuclear cells; strong Bax immunostaining was detected in dilated dentinal tubules close to the lesion, and Bcl-2 staining was weak in some dentin areas under the cavity or altogether absent. These findings suggest that both apoptosis pathways are activated in dental caries. Further studies are required to gain insights into its biomolecular mechanisms. PMID:26428882

  6. Non-functioning pituitary adenoma: immunohistochemical analysis of 85 cases.

    PubMed

    Mahta, Ali; Haghpanah, Vahid; Lashkari, Anahita; Heshmat, Ramin; Larijani, Bagher; Tavangar, Seyed Mohammad

    2007-01-01

    Pituitary adenomas without clinically active hypersecretion are summarized under the term non-functioning pituitary adenoma (NFPA). Since there are no specific serum markers, the differential diagnosis and treatment imply special difficulties. By using immunohistochemical methods we will have new insight into the nature and pathogenesis of these tumours. Ki-67 is a nuclear antigen detected by the monoclonal antibody MIB-1 and its labelling index (LI) is considered a marker of normal and abnormal cell proliferation. The aim of this study was to investigate the possible role of immunohistochemistry and MIB1-LI determination in NFPAs to predict tumoural behaviour and better management. In this clinicopathological study, 85 cases of NFPAs were analysed immunohistochemically. MIB1-LI was also determined in studied cases. Clinical presentation, treatment and follow-up data were also reviewed and the correlation between clinical and pathologic findings was established. Eighteen adenomas (21.2%) were immunoreactive to one or two adenohypophysial hormones of which 4 GH positive adenomas had aggressive behaviour (2 significant juxtasellar extensions and 2 recurrences). MIB-1 LI was more than 5% in only 5 cases including 2 invasive adenomas but with no evidence of recurrence. No significant statistical difference between clinical presentations in immunoreactive and non-immunoreactive NFPAs was observed except for unilateral temporal hemianopia which was more common in immunoreactive adenomas (P=0.022). NFPAs comprise several pathologically different types of tumours, some of which are potentially hormone producing, but some defects in hormone secretion or production of biologically inactive or insufficient amount of hormone may be the culprit in the lack of evidence of rising serum hormone levels. MIB-1 LI may be indicative of invasiveness but not a predictor of recurrence. Silent somatotropinomas may have more aggressive behaviour in comparison with other NFPAs. PMID

  7. TGF-ß isoforms in cancer: Immunohistochemical expression and Smad-pathway-activity-analysis in thirteen major tumor types with a critical appraisal of antibody specificity and immunohistochemistry assay validity.

    PubMed

    Riemenschneider, Markus J; Hirblinger, Maria; Vollmann-Zwerenz, Arabel; Hau, Peter; Proescholdt, Martin A; Jaschinski, Frank; Rothhammer-Hampl, Tanja; Wosikowski, Katja; Janicot, Michel; Leo, Eugen

    2015-09-29

    The literature on TGF-ß in cancer including data on the expression or activation of TGF-ß pathway components in specific tumors types is steadily growing. However, no systematic and uniform analysis exists reporting expression levels of the main TGF-ß pathway components across the most frequent tumor types. We used a standardized immunohistochemical assay investigating TGF-ß isoform expression and pathway activation across 13 different tumor types and corresponding non-neoplastic tissues. The study was performed on tissue microarrays allowing for the parallel analysis of a total of 1638 human tumor samples. TGF-ß1, TGF-ß2 and p-Smad2/3 were substantially expressed in multiple cancers widening the options for TGF-ß isoform directed therapies. Of note, TGF-ß antigens appear to be expressed in an individual manner pointing towards a need for patient preselection for TGF-β isoform specific treatment. Yet, a thorough investigation of antibody specificity and assay validity revealed that immunohistochemistry did not correlate with other detection methods on mRNA or protein level in all instances. As such, with the currently available means (i.e. antibodies tested) a stratification of patients within clinical trials for TGF-ß directed antisense therapies based upon TGF-β immunohistochemistry alone has to be interpreted with caution and should be carefully evaluated in combination with other parameters.

  8. Immunohistochemical analysis of growth mechanisms in juvenile nasopharyngeal angiofibroma.

    PubMed

    Schuon, Robert; Brieger, Jürgen; Heinrich, Ulf R; Roth, Yeduha; Szyfter, Witold; Mann, Wolf J

    2007-04-01

    Angiogenic factors are discussed to participate in growth and promotion of juvenile nasopharyngeal angiofibroma (JNA). However, only few data are available and mechanisms remain unclear. In the presented study we analysed the expression and subcellular distribution of several angiogenic growth factors and receptors potentially involved in JNA-growth and -vascularisation. In a retrospective, descriptive, multicenter-study, we analysed 13 formalin-fixed, paraffin-embedded or cryopreserved JNA-tumors (eleven primary tumors and two recurrent ones) after immunohistochemical staining. We used monoclonal antibodies specific for transforming growth factor beta 1 (TGF-beta(1)), basic fibroblast growth factor (bFGF), the VEGF-receptors 1 and -2 (FLT-1 and FLK-1), and the hypoxia inducible factor (Hif-1alpha). Data were compared to the vessel density. Quantitative analysis of staining intensities was performed by a computer assisted quantification technique. Endothelial and stromal compartments of the samples were analysed separately. Data were compared to vessel densities and patients data. The VEGF-Receptor-2 (FLK) was frequently unregulated in the stroma and endothelium of those samples with high vessel densities. Similarly, we observed high bFGF- and TGF-beta(1) levels in the stroma of strong vascularised samples. No correlations of expression levels to patients' data were found. The reported data support the concept of JNA-growth and -vascularisation driven by factors released from stromal fibroblasts. Therefore, inhibition of these factors might be beneficial for the therapy of inoperable JNA.

  9. Immunohistochemical analysis of cartilage-derived retinoic acid-sensitive protein (CD-RAP)/melanoma inhibitory activity (MIA) in murine, canine, bovine and equine cerebrospinal tissues.

    PubMed

    Tokunaga, Satoshi; Fujiki, Makoto; Yabuki, Akira; Misumi, Kazuhiro

    2012-04-01

    Cartilage-derived retinoic acid-sensitive protein (CD-RAP)/melanoma inhibitory activity (MIA), which appears abundantly in hypertrophic cartilage at the stage of endochondral ossification, is also detected in cerebrospinal fluid (CSF) following spinal cord injury. In this study, the localization of the CD-RAP/MIA molecule in normal tissues of the spine and brain obtained from mice, rats, dogs, cattle and horses was examined using immunohistochemistry with a specific antibody. The positive signals of CD-RAP/MIA were found at nerve cells in the spinal cords of all species and were especially strong at cerebellar Purkinje cells. The results suggested that CD-RAP/MIA included in normal cerebrospinal tissues could be a biomarker associated with tissue injuries, as the molecules might flow into the CSF.

  10. Canine nail bed keratoacanthoma diagnosed by immunohistochemical analysis

    PubMed Central

    Yoo, Chang-Bum; Kim, Dae-Hyun; Lee, A-Jin; Suh, Hyun-Jung; Yoo, Saejong; Sur, Jung-Hyang; Eom, Ki Dong

    2015-01-01

    A 10-year-old, Shih Tzu dog was presented with an enlarged, curled 2nd nail in the left forelimb. Digital amputation was performed and the mass was diagnosed as a nail bed keratoacanthoma (infundibular keratinizing acanthoma) histopathologically. There was no recurrence postoperatively. This is the first case report of a canine nail bed keratoacanthoma diagnosed by histologic and immunohistochemical examination including Ki-67 and p53 expression. PMID:26538676

  11. [Use of computer-assisted image analysis for the detection of immunohistochemical receptor status in breast cancer. Immunohistochemical densitometry receptor analysis--IRDA].

    PubMed

    Mink, D; Bonkhoff, H; Herth, G; Schmidt, W

    1992-10-01

    A method for the evaluation of the immunohistochemical staining score of steroid receptors in breast cancer using computer-based image analysis is presented. The optical density of tumour cell nuclei was measured using kryostat sections and, using a control section as reference, an objective threshold for specific staining was defined. In intensively stained sections, a good correlation with visual scoring was achieved, but in 9 out of 16 sections, which were regarded as receptor-negative by visual scoring, a specific receptor-positive staining was demonstrated by image analysis. Next to the percentage of positive cells (PP) and the mean staining intensity (SI), a histogram of the receptor concentration in positive nuclei was calculated. The method allows, besides an objective evaluation and documentation, an exact definition of a threshold for specific receptor measurement and furthermore shows the pattern of receptor concentration in the tumour cell nuclei. Hence, the method could contribute towards a more widespread use of immunohistochemical receptor analysis.

  12. Immunohistochemical detection of active transforming growth factor-beta in situ using engineered tissue

    NASA Technical Reports Server (NTRS)

    Barcellos-Hoff, M. H.; Ehrhart, E. J.; Kalia, M.; Jirtle, R.; Flanders, K.; Tsang, M. L.; Chatterjee, A. (Principal Investigator)

    1995-01-01

    The biological activity of transforming growth factor-beta 1 (TGF-beta) is governed by dissociation from its latent complex. Immunohistochemical discrimination of active and latent TGF-beta could provide insight into TGF-beta activation in physiological and pathological processes. However, evaluation of immunoreactivity specificity in situ has been hindered by the lack of tissue in which TGF-beta status is known. To provide in situ analysis of antibodies to differentiate between these functional forms, we used xenografts of human tumor cells modified by transfection to overexpress latent TGF-beta or constitutively active TGF-beta. This comparison revealed that, whereas most antibodies did not differentiate between TGF-beta activation status, the immunoreactivity of some antibodies was activation dependent. Two widely used peptide antibodies to the amino-terminus of TGF-beta, LC(1-30) and CC(1-30) showed marked preferential immunoreactivity with active TGF-beta versus latent TGF-beta in cryosections. However, in formalin-fixed, paraffin-embedded tissue, discrimination of active TGF-beta by CC(1-30) was lost and immunoreactivity was distinctly extracellular, as previously reported for this antibody. Similar processing-dependent extracellular localization was found with a neutralizing antibody raised to recombinant TGF-beta. Antigen retrieval recovered cell-associated immunoreactivity of both antibodies. Two antibodies to peptides 78-109 showed mild to moderate preferential immunoreactivity with active TGF-beta only in paraffin sections. LC(1-30) was the only antibody tested that discriminated active from latent TGF-beta in both frozen and paraffin-embedded tissue. Thus, in situ discrimination of active versus latent TGF-beta depends on both the antibody and tissue preparation. We propose that tissues engineered to express a specific form of a given protein provide a physiological setting in which to evaluate antibody reactivity with specific functional forms of a

  13. Hyperplastic corneal pannus: an immunohistochemical analysis and review.

    PubMed

    Jakobiec, Frederick A; Stacy, Rebecca C; Mendoza, Pia R; Chodosh, James

    2014-01-01

    An exuberant corneal pannus usually develops in adults with a history of surgery or trauma in the anterior central stroma and appears as a glistening, vascularized, moderately elevated, well circumscribed white nodule. We describe a 78-year-old woman with such a pannus, which in the past has typically been referred to as keloidal or hypertrophic. The involved eye had only light perception, and she underwent a penetrating keratoplasty that improved her vision to 20/100. Histopathologic and immunohistochemical evaluations of a the specimen disclosed a reactive spindle cell stromal proliferation of myofibroblasts that were smooth muscle actin positive with a low Ki67 proliferation index. Desmin, caldesmon, and calponin were negative, in keeping with the incomplete myofilamentary differentiation of a myofibroblast. There was a generous admixture of CD68/163-positive histiocytes and dispersed C3/5-positive T-lymphocytes. An absence of CD138- and IgG4-positive plasma cells ruled out an IgG4-related disease. For a lesion to be keloidal, the collagen must have a thick hyaline character, sharp edges, and a sparsity of intervening cells and vessels. A hypertrophic pannus would be composed of large swollen cells not necessarily increased in number. We therefore recommend adoption of the term hyperplastic for lesions like that described here because of the obvious increase in cellularity from proliferating myofibroblasts and the lack of true keloidal collagen.

  14. Immunohistochemical analysis of steroid hormone receptors in nasopharyngeal angiofibromas.

    PubMed

    Gatalica, Z

    1998-05-15

    Juvenile nasopharyngeal angiofibroma arises almost exclusively in pubertal and adolescent men and has potentially aggressive behavior with a spread into adjoining sinuses and bone destruction. It is classically being regarded as an androgen hormone-dependent tumor, but no in situ evaluation of androgen receptors has been done. The author has examined eight nasopharyngeal angiofibromas (six primary and two recurrent tumors) for the expression of androgen receptor (AR), estrogen receptor (ER) and progesterone receptor (PR) using immunohistochemical methods and compared those results with a sex- and age-matched control group consisting of eight samples of nasal turbinates. No ER or PR were found in any of the tumor components, nor have they been detected in control nasal turbinates. Angiofibromas were characterized by variable weak (+) nuclear androgen receptor immunoreactivity found in a minority of endothelial and stromal cells, similar to the normal turbinates. These results argue against the significant role of androgen receptor in the growth of nasal angiofibromas and corroborate previous observations of an unpredictable response of these neoplasms to antiandrogen therapy.

  15. Immunohistochemical analysis of retroperitoneal Müllerian cyst.

    PubMed

    Konishi, Eiichi; Nakashima, Yasuaki; Iwasaki, Takeki

    2003-02-01

    Retroperitoneal cysts are uncommon diseases, and benign nonneoplastic Müllerian cysts are extremely rare among the known cases. We report a case of a 35-year-old woman with a retroperitoneal Müllerian cyst with the tubal type of epithelium. The patient presented with a large (20 cm in diameter), palpable abdominal mass. This multilocular cystic mass was resected from the retroperitoneum between the descending colon and the left renal fascia. Histologically, it was lined by monolayered low-cuboidal to columnar cells without atypia that resembled tubal epithelium, including cilia. Loose fibrous tissue and incomplete smooth muscle bundles were identified beneath the epithelium of the lining. Immunohistochemical tests showed that the lining cells were strongly positive for cytokeratins (CKs) (polyclonal, 7, 18, CAM 5.2, AE1/AE3), epithelial membrane antigen, cancer antigen 125, progesterone receptor, and estrogen receptor. The lining cells were also occasionally weakly positive for CK5/6. They tested negative for CK20, carcinoembryonic antigen, calretinin, and CD 10.

  16. Radiographic and immunohistochemical analysis of leukocyte adhesion deficiency in Holstein heifers.

    PubMed

    Nagahata, H; Taniyama, H; Izumisawa, Y; Kotani, T; Noda, H; Kociba, G J

    1995-10-01

    Radiographic and immunohistochemical analyses were performed in two Holstein heifers with leukocyte adhesion deficiency (BLAD). Severe bone resorption, osteolysis and severe progressive periodontitis in submandibula due to dysfunction of leukocytes in heifers affected with BLAD were demonstrated by radiographic examination. Immunohistochemical analysis of lymph nodes using anti-CD18 monoclonal antibody demonstrated that CD18-positive cells were not found on those from a heifer affected with BLAD, whereas CD18-positive cells were clearly present in lymph nodes from a clinically normal heifer. These characteristic findings support the importance of adherence-dependent leukocyte functions in host defense. PMID:8548695

  17. Immunohistochemical Analysis of the Structure of Injured Peripheral Nerve Neuroma after Electrosurgical Welding Intervention.

    PubMed

    Korsak, A V; Chaikovskii, Yu B

    2015-10-01

    Immunohistochemical analysis of changes in neuroma after surgical treatment of damaged peripheral nerve with the use of high frequency electrosurgical device for high frequency current welding of soft tissues was carried out. No adverse effects of this technology and the bipolar instrument on degeneration and regeneration of damaged nerve stem were detected.

  18. Immunohistochemical analysis of carbohydrate antigens in chronic inflammatory gastrointestinal diseases.

    PubMed

    Kobayashi, Motohiro; Nakayama, Jun

    2010-01-01

    Over the last four decades, immunohistochemistry (IHC) has become an invaluable technique to detect antigens in tissue sections. Compared to Western blotting analysis, IHC is advantageous in determining histological distribution and localization of the antigen. Another advantage, if one can access human formalin-fixed paraffin-embedded (FFPE) blocks of disease tissues, is that IHC makes it possible to analyze diseases retrospectively from archived pathological tissue specimens. In this chapter, we describe protocols used for both conventional and multiple immunostainings using FFPE tissue sections, which have been used for quantitative analysis of high endothelial venule (HEV)-like vessels and lymphocyte subsets attached to HEV-like vessels in our studies of chronic inflammatory gastrointestinal diseases. We also describe in detail a protocol using an L-selectinIgM chimera in situ binding assay on FFPE tissue sections for functional detection of L-selectin ligand carbohydrates expressed on HEV-like vessels. After presenting each protocol, we provide practical examples for its use obtained from our studies.

  19. Oncocytic lipoadenoma of the parotid gland: Immunohistochemical and cytogenetic analysis.

    PubMed

    Ilie, Marius; Hofman, Véronique; Pedeutour, Florence; Attias, Rita; Santini, Joseph; Hofman, Paul

    2010-01-15

    Salivary gland oncocytic lipoadenoma is an exceptional benign tumor composed of mature adipose tissue associated with a mixture of oncocytes. We report a case of oncocytic lipoadenoma showing sebaceous differentiation, and provide a cytogenetic analysis, which has not yet been described. A 64-year-old male developed a left parotid gland, well-encapsulated tumor measuring 3.5 x 3 cm(2), showing mature fat cells associated with oncocytic changes of epithelial components. Immunohistochemistry showed a dual epithelial population with ductal (positivity for AE1/AE3, CK19, CK7 antibodies) and basal-cell (positivity for p63, CK14, CK5,6 antibodies) differentiation in oncocytic areas. Moreover, oncocytic cells were stained with anti-alpha-1 antichymotrypsin antibody and phosphotungstic acid-hematoxylin staining. Molecular cytogenetic analysis showed a translocation t(12;14), resulting in structural rearrangement of the region framing the HMGA2 gene at 12q14.3. Such alterations in HMGA2 have been described in both lipomas and pleomorphic adenomas of the salivary glands.

  20. Lipomatous Change in Uveal Melanoma: Histopathological, Immunohistochemical and Cytogenetic Analysis

    PubMed Central

    Yavuzyigitoglu, Serdar; Kilic, Emine; Vaarwater, Jolanda; de Klein, Annelies; Paridaens, Dion; Verdijk, Robert M.

    2016-01-01

    Purpose The aim of this study was to describe a case of lipomatous change in uveal melanoma. Procedures The patient presented with a 2-year history of blurry vision. A full examination of the right eye revealed a dome-shaped pigmented subretinal mass in the choroid with a thickness of 9 mm and a diameter of 15 mm. The eye was enucleated and prepared for histopathologic, genetic and molecular investigation. Results Histopathology revealed a small circumscribed area consisting of mature adipocytic appearing cells with abundant clear cytoplasm and small peripheral flattened nuclei within a spindle-cell melanoma of the uvea. The cytoplasm of the adipocytic cells stained negative for periodic acid-Schiff and Alcian blue and positive for Melan-A, HMB-45 and tyrosinase, confirming melanocytic lineage. Fluorescence in situ hybridization analysis confirmed trisomy of chromosome 6p22 and disomy of chromosome 3p13 in the nuclei of both the tumor spindle type B cells and in the nuclei of lipomatous tumor cells. Conclusions Lipomatous change can be added to the many histopathologic faces of uveal melanoma. To our knowledge, this is the first report of lipomatous change in uveal melanoma performed with cytogenetic investigations. PMID:27239451

  1. Immunohistochemical analysis of immune response in breast cancer and melanoma patients after laser immunotherapy

    NASA Astrophysics Data System (ADS)

    Nordquist, Robert E.; Bishop, Shelly L.; Ferguson, Halie; Vaughan, Melville B.; Jose, Jessnie; Kastl, Katherine; Nguyen, Long; Li, Xiaosong; Liu, Hong; Chen, Wei R.

    2011-03-01

    Laser immunotherapy (LIT) has shown great promise in pre-clinical studies and preliminary clinical trials. It could not only eradicate treated local tumors but also cause regression and elimination of untreated metastases at distant sites. Combining a selective photothermal therapy with an active immunological stimulation, LIT can induce systemic anti-tumor immune responses. Imiquimod (IMQ), a toll-like receptor agonist, was used for the treatment of late-stage melanoma patients and glycated chitosan (GC), a biological immunological modulator, was used for the treatment of late-stage breast cancer patients, in combination of irradiation of a near-infrared laser light. To observe the immunological changes before and after LIT treatment, the pathological tissues of melanoma and breast cancer patients were processed for immunohistochemical analysis. Our results show that LIT changed the expressions of several crucial T cell types. Specifically, we observed significant decreases of CD3+ T-cells and a significant increase of CD4+,CD8+, and CD68+ T-cells in the tumor samples after LIT treatment. While not conclusive, our study could shed light on one the possible mechanisms of anti-tumor immune responses induced by LIT. Further studies will be conducted to identify immunological biomarkers associated with LIT-induced clinical response.

  2. Clinical, dermoscopic and immunohistochemical assessment of actinic keratoses and evaluation of the effectiveness of diclofenac therapy with immunohistochemical analysis.

    PubMed

    Çayirli, Mutlu; Köse, Osman; Demiriz, Murat

    2013-07-01

    Actinic keratoses (AKs) is a keratinocytic neoplasm that typically develops on the face of elderly patients. Little is known regarding the clinical, dermatoscopic and immunohistochemical assessments of AK using topical diclofenac therapy. We sought to determine these assessments and evaluate the efficacy of topical diclofenac gel in AK. In this prospective, open-label study, 44 patients with 66 AKs were treated for 12 weeks with topically applied diclofenac (3% gel in 2.5% hyaluronic acid). Immunohistopathologic analyses were performed before and after diclofenac treatment using epidermal stem cell markers such as Cytokeratin 15 (CK15), Cytokeratin 19 (CK19) and p63, in addition to proliferation markers (Bcl-2, Ki-67). Diclofenac gel was found to be effective in AK, including the hyperkeratotic type. Surprisingly, complete remission was observed at a significantly higher rate in Grade 3 lesions (p = 0.017). However, imunohistochemical and histopathologic examinations revealed that 12-week treatment periods may not be sufficient to fully cure AK. The immunohistochemical analyses revealed no change in the expression levels of CK15, CK19 and Bcl-2 following diclofenac therapy. However, the expression of Ki-67 (p = 0.042) and p63 (p = 0.030) exhibited a significant decrease after therapy. Dermatoscopy is an effective method for diagnosis of AK, and topical diclofenac sodium gel was found as an effective additional treatment modality. Since positive histopathological findings were detected in some patients even with significant remission, a 12-week treatment period should be extended even in patients presenting with positive clinical response. Importantly, anti-proliferative effects of diclofenac were demonstrated by decreased Ki-67 and p63 expression levels.

  3. Non-sebaceous lymphadenoma of the parotid gland: immunohistochemical study and DNA ploidy analysis.

    PubMed

    Gallego, Lorena; Junquera, Luis; Fresno, Manuel F

    2009-04-01

    Nonsebaceous lymphadenoma (NSL) is an unusual benign salivary gland tumor characterized by a predominant lymphoid background, dense lymphoid infiltrate, and absence of sebaceous differentiation. To our knowledge, only 10 previous cases have been reported in the literature. We report an additional case of NSL arising in the parotid gland in 58-year-old female patient. The extensive immunohistochemical investigation of the tumor revealed the presence of both luminal and myoepithelial cells. DNA analysis for flow cytometry was performed. The histogram presented a single peak in the G0-G1 area. The tumor was considered as being DNA diploid.

  4. Extracellular collagenous spherules in salivary gland tumors. Immunohistochemical analysis of laminin and various types of collagen.

    PubMed

    Skalova, A; Leivo, I

    1992-06-01

    Collagenous spherulosis is a benign breast lesion involving lobular acini and ductules and containing eosinophilic spherules measuring up to 100 microns in diameter. We present an immunohistochemical analysis of similar collagen-rich spherules that are also found in salivary gland tumors. These collagenous spherules contain varying amounts of acidic mucins, elastin, basement membrane proteins including type IV collagen and laminin, and considerable amounts of interstitial collagen types I and III. Types II and VI collagen were not detected in collagenous spherules of salivary gland tumors. The cells surrounding these collagenous spherules expressed muscle actin, S100 protein, vimentin, and cytokeratins 8, 18, and 19, indicating that these cells have myoepithelial characteristics.

  5. Reevaluation and reclassification of resected lung carcinomas originally diagnosed as squamous cell carcinoma using immunohistochemical analysis.

    PubMed

    Kadota, Kyuichi; Nitadori, Jun-ichi; Rekhtman, Natasha; Jones, David R; Adusumilli, Prasad S; Travis, William D

    2015-09-01

    Currently, non-small cell lung carcinomas are primarily classified by light microscopy. However, recent studies have shown that poorly differentiated tumors are more accurately classified by immunohistochemistry. In this study, we investigated the use of immunohistochemical analysis in reclassifying lung carcinomas that were originally diagnosed as squamous cell carcinoma. Tumor slides and blocks were available for histologic evaluation, and tissue microarrays were constructed from 480 patients with resected lung carcinomas originally diagnosed as squamous cell carcinoma between 1999 and 2009. Immunohistochemical analyses for p40, p63, thyroid transcription factor-1 (TTF-1; clones SPT24 and 8G7G3/1), napsin A, chromogranin A, synaptophysin, and CD56 were performed. Staining intensity (weak, moderate, or strong) and distribution (focal or diffuse) were also recorded. Of all, 449 (93.5%) patients were confirmed as having squamous cell carcinomas; the cases were mostly diffusely positive for p40 and negative for TTF-1 (8G7G3/1). Twenty cases (4.2%) were reclassified as adenocarcinoma, as they were positive for TTF-1 (8G7G3/1 or SPT24) with either no or focal p40 expression, and all of them were poorly differentiated with squamoid morphology. In addition, 1 case was reclassified as adenosquamous carcinoma, 4 cases as large cell carcinoma, 4 cases as large cell neuroendocrine carcinoma, and 2 cases as small cell carcinoma. In poorly differentiated non-small cell lung carcinomas, an accurate distinction between squamous cell carcinoma and adenocarcinoma cannot be reliably determined by morphology alone and requires immunohistochemical analysis, even in resected specimens. Our findings suggest that TTF-1 8G7G3/1 may be better suited as the primary antibody in differentiating adenocarcinoma from squamous cell carcinoma.

  6. Activation of Mammalian target of rapamycin in canine mammary carcinomas: an immunohistochemical study.

    PubMed

    Delgado, L; Gärtner, F; Dias Pereira, P

    2015-01-01

    Mammalian target of rapamycin (mTOR) is a serine-threonine kinase involved in cell growth, proliferation and survival. Activation of mTOR has been reported in various tumour types, including human breast cancer; however, the expression of mTOR in canine mammary tumours has not been examined. In the present study, expression of the activated form of mTOR (phospho-mTOR [p-mTOR]) was examined immunohistochemically in five normal canine mammary glands, 45 canine mammary carcinomas and their corresponding metastatic lesions (n = 15). Phospho-mTOR was not expressed in normal canine mammary tissue, but cytoplasmic labelling was observed in 78% of canine mammary carcinomas. Two carcinomas had both cytoplasmic and nuclear labelling. No significant relationship was found between p-mTOR cytoplasmic expression and histological type or grading of carcinomas, degree of tubular formation, anisokaryosis, mitotic activity or lymph node metastasis. In all except one case, the expression pattern of p-mTOR in lymph node metastases was similar or decreased when compared with the primary lesion. The findings suggest that p-mTOR is involved in mammary carcinogenesis in dogs. However, p-mTOR cytoplasmic expression does not appear to be a prognostic indicator in canine mammary carcinomas, which may be related to its subcellular location in the neoplastic cells. Canine mammary tumours may provide a model for the development of innovative medical strategies involving mTOR inhibitors in human breast cancer. PMID:25670666

  7. Immunohistochemical identification and quantitative analysis of cytoplasmic Cu/Zn superoxide dismutase in mouse organogenesis

    PubMed Central

    Yon, Jung-Min; Baek, In-Jeoung; Lee, Se-Ra; Kim, Mi-Ra; Lee, Beom Jun; Yun, Young Won

    2008-01-01

    Cytoplasmic Cu/Zn superoxide dismutase (SOD1) is an antioxidant enzyme that converts superoxide to hydrogen peroxide in cells. Its spatial distribution matches that of superoxide production, allowing it to protect cells from oxidative stress. SOD1 deficiencies result in embryonic lethality and a wide range of pathologies in mice, but little is known about normal SOD1 protein expression in developing embryos. In this study, the expression pattern of SOD1 was investigated in post-implantation mouse embryos and extraembryonic tissues, including placenta, using Western blotting and immunohistochemical analyses. SOD1 was detected in embryos and extraembryonic tissues from embryonic day (ED) 8.5 to 18.5. The signal in embryos was observed at the lowest level on ED 9.5-11.5, and the highest level on ED 17.5-18.5, while levels remained constant in the surrounding extraembryonic tissues during all developmental stages examined. Immunohistochemical analysis of SOD1 expression on ED 13.5-18.5 revealed its ubiquitous distribution throughout developing organs. In particular, high levels of SOD1 expression were observed in the ependymal epithelium of the choroid plexus, ganglia, sensory cells of the olfactory and vestibulocochlear epithelia, blood cells and vessels, hepatocytes and hematopoietic cells of the liver, lymph nodes, osteogenic tissues, and skin. Thus, SOD1 is highly expressed at late stages of embryonic development in a cell- and tissue-specific manner, and can function as an important antioxidant enzyme during organogenesis in mouse embryos. PMID:18716442

  8. Immunohistochemical analysis of myenteric ganglia and interstitial cells of Cajal in ulcerative colitis

    PubMed Central

    Bernardini, Nunzia; Segnani, Cristina; Ippolito, Chiara; De Giorgio, Roberto; Colucci, Rocchina; Faussone-Pellegrini, Maria Simonetta; Chiarugi, Massimo; Campani, Daniela; Castagna, Maura; Mattii, Letizia; Blandizzi, Corrado; Dolfi, Amelio

    2012-01-01

    Abstract Ulcerative colitis (UC) is an inflammatory bowel disease with alterations of colonic motility, which influence clinical symptoms. Although morpho-functional abnormalities in the enteric nervous system have been suggested, in UC patients scarce attention has been paid to possible changes in the cells that control colonic motility, including myenteric neurons, glial cells and interstitial cells of Cajal (ICC). This study evaluated the neural-glial components of myenteric ganglia and ICC in the colonic neuromuscular compartment of UC patients by quantitative immunohistochemical analysis. Full-thickness archival samples of the left colon were collected from 10 patients with UC (5 males, 5 females; age range 45–62 years) who underwent elective bowel resection. The colonic neuromuscular compartment was evaluated immunohistochemically in paraffin cross-sections. The distribution and number of neurons, glial cells and ICC were assessed by anti-HuC/D, -S100β and -c-Kit antibodies, respectively. Data were compared with findings on archival samples of normal left colon from 10 sex- and age-matched control patients, who underwent surgery for uncomplicated colon cancer. Compared to controls, patients with UC showed: (i) reduced density of myenteric HuC/D+ neurons and S100β+ glial cells, with a loss over 61% and 38%, respectively, and increased glial cell/neuron ratio; (ii) ICC decrease in the whole neuromuscular compartment. The quantitative variations of myenteric neuro-glial cells and ICC indicate considerable alterations of the colonic neuromuscular compartment in the setting of mucosal inflammation associated with UC, and provide a morphological basis for better understanding the motor abnormalities often observed in UC patients. PMID:21426484

  9. An immunohistochemical study of bizarre neoplastic cells in pleomorphic adenoma: its cytological nature and proliferative activity.

    PubMed

    Takeda, Y

    1999-11-01

    The cytological nature and proliferative activity of bizarre neoplastic cells, widely scattered in pleomorphic adenomas of salivary gland origin were studied. Pleomorphic adenomas containing numerous bizarre neoplastic cells were found in four cases, and were equal to 2.9% of all pleomorphic adenomas examined. All four cases presented as well-circumscribed, firm masses measuring less than 1.5 cm in size, located in the palate, and were of 7 months to 4 years duration. Histopathologically, these pleomorphic adenomas were cell rich type, and were well demarcated from surrounding tissues, although their fibrous capsules were partially defective. In addition to characteristic histopathological findings of pleomorphic adenoma, numerous neoplastic cells with bizarre appearance were scattered throughout the lesion, excepting for tubuloductal structures. These bizarre neoplastic cells had irregular-shaped and large nuclei with or without hyperchromatism, although their nucleoli were small and mitotic figures were few. Furthermore, there were many multinucleated giant cells, some of which showed multilobulated nuclei. Neither necrosis nor infarct was seen in the tumors. Immunohistochemically, bizarre neoplastic cells scattered in solid-proliferating areas and myxoid areas were neoplastic myoepithelial cells in nature. There was no statistical significance of MIB-1 labeling indices between pleomorphic adenomas with bizarre neoplastic cells and usual pleomorphic adenomas. The p53 labeling indices were quite low. Although the benign nature of pleomorphic adenomas with numerous bizarre neoplastic cells and hypercellularity, distinguishing such pleomorphic adenomas from various stages of malignant transformation in pleomorphic adenomas and other carcinomas should be made by histological section of submitted biopsy specimen or aspirated content for cytological diagnosis. The present paper suggests that the term 'bizarre cell pleomorphic adenoma' is an appropriate name for this

  10. Effect of light-emitting diode (ʎ 627 nm and 945 nm ʎ) treatment on first intention healing: immunohistochemical analysis.

    PubMed

    Kerppers, Ivo Ilvan; de Lima, Carlos José; Fernandes, Adriana Barrinha; Villaverde, Antonio Balbin

    2015-01-01

    Collagen I is not only responsible for maintaining the integrity of most tissues due to its mechanical properties, but also for its active participation in the functionality of tissues because of its interaction with cells present in the extracellular matrix. The synthesis of collagen begins with tissue injury and remains until the end of the healing process. The use of non-coherent light for healing processes is still understudied. This procedure stands out as a biostimulation method for tissue repair, which increases local circulation, cell proliferation, and collagen synthesis. This study sought to quantify collagen I in the healing process after the treatment of wounds with the light-emitting diode (LED) treatment. The histologic analysis with tissue samples stained with picrosirius red showed a statistical difference between the positive controls, LED 627 and LED 945 nm groups; the group treated with LED 627 nm showed a predominance of mature collagen. The immunohistochemical analysis showed a statistically significant high concentration of collagen I in the LED 945 nm group. The irradiation of wounds with the higher wavelength (945 nm) used in the study produced the best activity of collagen I formation in experimental model.

  11. "Mucin"-secreting papillary renal cell carcinoma: clinicopathological, immunohistochemical, and molecular genetic analysis of seven cases.

    PubMed

    Pivovarcikova, Kristyna; Peckova, Kvetoslava; Martinek, Petr; Montiel, Delia Perez; Kalusova, Kristyna; Pitra, Tomas; Hora, Milan; Skenderi, Faruk; Ulamec, Monika; Daum, Ondrej; Rotterova, Pavla; Ondic, Ondrej; Dubova, Magdalena; Curik, Romuald; Dunatov, Ana; Svoboda, Tomas; Michal, Michal; Hes, Ondrej

    2016-07-01

    Mucin and mucin-like material are features of mucinous tubular and spindle renal cell carcinoma (MTS RCC) but are rarely seen in papillary renal cell carcinoma (PRCC). We reviewed 1311 PRCC and identified 7 tumors containing extracellular and/or intracellular mucinous/mucin-like material (labeled as PRCCM). We analyzed these using morphological, histochemical, immunohistochemical, and molecular genetic methods (arrayCGH, FISH). Clinical data were available for six of the seven patients (five males and one female, age range 61-78 years). Follow-up was available for four patients (2-4 years); one patient died of widespread metastases. Tumor size ranged from 3 to 5 cm (mean 3.8). Of all cases, histological architecture showed a predominantly papillary pattern. Mucin or mucin-like was extracellular in one, intracellular in three, and both intra/extracellular in three cases. All tumors were positive for AMACR, vimentin, and OSCAR, while CK7 was positive in four. Mucicarmine stain was positive in all cases, PAS in six and Alcian blue in three cases. Five tumors were positive for MUC 1, but none were positive for MUC 2, MUC 4, or MUC 6. In only four cases, genetic analysis could be performed. Gain of chromosomes 7 and 17 was found in two cases; gain of 17 only was found in one case. Loss of heterozygosity of 3p was found in one case together with polysomy of chromosomes 7 and 17. No abnormalities of VHL, fumarate dehydrogenase, and TFE3 genes were detected. We conclude that PRCCM is a rare but challenging subtype of RCC that deserves to be further studied. In all the tumors, the mucin-like material was found in those stained with mucicarmin, but other conventional and immunohistochemical stains did not reveal consistent features of a single mucin. The molecular-genetic profile of these tumors was most consistent with that of typical papillary RCC, although one case had mixed genetic features of papillary and clear RCC. PRCCM has metastatic potential, as evidenced by

  12. Immunohistochemical expression of the oncogenic molecules active Stat3 and survivin in benign and malignant salivary gland tumors

    PubMed Central

    Nikitakis, Nikolaos G.; Scheper, Mark A.; Papanicolaou, Vasileios S.; Sklavounou, Alexandra; Sauk, John J.

    2009-01-01

    Objective Signal transducer and activator of transcription 3 (Stat3) and survivin have been shown to exert oncogenic effects in various human neoplasms. The purpose of this study was to evaluate the expression of the tyrosine phosphorylated (active) Stat3 and survivin in various benign and malignant salivary gland tumors (SGTs). Study design Eighty-six SGTs (65 malignant and 21 benign tumors of various histopathologic subtypes) were immunohistochemically stained with anti-survivin or anti-phosphorylated tyrosine-705 (p-tyr) Stat3 antibodies. Immunohistochemical reactivity was graded in a semi-quantitative manner; a combined score of immunohistochemical positivity (0–6) was calculated for each tumor by adding the individual scores for percentage of tumor cells (0–3) and intensity of staining (0–3). Results Survivin was immunohistochemically detected in all studied benign and malignant SGTs; p-tyr Stat3 was also detected in the majority (91%) of SGTs. The average combined scores for survivin and p-tyr Stat3 immunohistochemical expression in the studied malignant SGTs was 4.40 and 3.35, respectively; the corresponding combined scores for survivin and p-tyr Stat3 in the studied benign SGTs were 4.37 and 3.22, respectively. No statistically significant differences (p>0.05) in p-tyr Stat3 or survivin expression were detected between the benign and malignant groups, or among the various examined histopathological subtypes of SGTs. In contrast, normal salivary gland elements in the vicinity of the studied tumors revealed only weak and focal survivin or p-tyr Stat3 immunoreactivity, mainly localized to ductal and mucous cells. Conclusions Our data indicate an almost universal expression of activated Stat3 and survivin in benign and malignant SGTs. Considering the well-established proliferative and anti-apoptotic properties of these molecules and their functional interrelationship, selective targeting techniques against Stat3 and/or survivin may represent promising

  13. Immunohistochemical analysis of p53 and ras p21 expression in colorectal adenomas and early carcinomas.

    PubMed

    Ieda, S; Watatani, M; Yoshida, T; Kuroda, K; Inui, H; Yasutomi, M

    1996-01-01

    To further investigate whether multiple genetic changes are involved in the development of colorectal cancer, we performed an immunohistochemical analysis of p53 and ras p21 protein expression in 139 specimens of colorectal adenoma with varying degrees of dysplasia, 57 specimens of early cancer with an adenomatous component, and 12 specimens of superficial early cancer without any adenomatous component. Positive p53 staining was found in 15% of the adenomas with moderate dysplasia and in 42% of the adenomas with severe dysplasia or intramucosal carcinoma (IMCA). Positive immunostaining of p53 was observed to be significantly correlated with the degree of dysplasia and the depth of invasion, as was the expression of ras p21. However, a closer correlation was observed with the increasing size of the adenomas. Furthermore, p53 staining was positive in 42% of the 12 superficial early cancer specimens, while ras staining was positive in only 1 specimen (8%). These results indicate that p53 gene overexpression may play some biological role in both the adenoma-to-carcinoma sequence and in de novo cancer development, whereas ras p21 expression may not be as involved in de novo cancer development as in the malignant conversion of colorectal adenomas.

  14. Immunohistochemical Expression of Matrix Metalloproteinase-7 in Human Colorectal Adenomas Using Specified Automated Cellular Image Analysis System: A Clinicopathological Study

    PubMed Central

    Qasim, Ban J.; Ali, Hussam H.; Hussein, Alaa G.

    2013-01-01

    Background/Aim: To evaluate the immunohistochemical expression of matrix metalloproteinase-7 (MMP-7) in colorectal adenomas, and to correlate this expression with different clinicopathological parameters. Patients and Methods: The study was retrospectively designed. Thirty three paraffin blocks from patients with colorectal adenoma and 20 samples of non-tumerous colonic tissue taken as control group were included in the study. MMP-7 expression was assessed by immunohistochemistry method. The scoring of immunohistochemical staining was conducted utilizing a specified automated cellular image analysis system (Digimizer). Results: The frequency of positive immunohistochemical expression of MMP-7 was significantly higher in adenoma than control group (45.45% versus 10%) (P value < 0.001). Strong MMP-7 staining was mainly seen in adenoma cases (30.30%) in comparison with control (0%) the difference is significant (P < 0.001). The three digital parameters of MMP-7 immunohistochemical expression (Area (A), Number of objects (N), and intensity (I)) were significantly higher in adenoma than control. Mean (A and I) of MMP-7 showed a significant correlation with large sized adenoma (≥ 1cm) (P < 0.05), also a significant positive correlation of the three digital parameters (A, N, and I) of MMP-7 expression with villous configuration and severe dysplasia in colorectal adenoma had been identified (P < 0.05). Conclusion: MMP-7 plays an important role in the growth and malignant conversion of colorectal adenomas as it is more likely to be expressed in advanced colorectal adenomatous polyps with large size, severe dysplasia and villous histology. The use of automated cellular image analysis system (Digmizer) to quantify immunohistochemical staining yields more consistent assay results, converts semi-quantitative assay to a truly quantitative assay, and improves assay objectivity and reproducibility. PMID:23319034

  15. A Detailed Immunohistochemical Analysis of a Large Series of Cervical and Vaginal Gastric-type Adenocarcinomas.

    PubMed

    Carleton, Claire; Hoang, Lien; Sah, Shatrughan; Kiyokawa, Takako; Karamurzin, Yevgeniy S; Talia, Karen L; Park, Kay J; McCluggage, W Glenn

    2016-05-01

    Adenocarcinomas exhibiting gastric differentiation represent a recently described and uncommon subtype of non-human papillomavirus (HPV)-related cervical adenocarcinoma. They comprise a spectrum from a well-differentiated variant (adenoma malignum/mucinous variant of minimal deviation adenocarcinoma) to a more poorly differentiated overtly malignant form, generally referred to as gastric-type adenocarcinoma. Rarely, such tumors have also been described as primary vaginal neoplasms. Gastric-type adenocarcinomas exhibit considerable morphologic overlap with adenocarcinomas originating outside the female genital tract, especially mucinous adenocarcinomas arising in the pancreas and biliary tract. Moreover, they often metastasize to unusual sites, such as the ovary and peritoneum/omentum, where they can be mistaken for metastatic adenocarcinomas from other, nongynecologic sites. There is little information regarding the immunophenotype of gastric-type adenocarcinomas, and knowledge of this is important to aid in the distinction from other adenocarcinomas. In this study, we undertook a detailed immunohistochemical analysis of a large series of cervical (n=45) and vaginal (n=2) gastric-type adenocarcinomas. Markers included were cytokeratin (CK)7, CK20, CDX2, carcinoembryonic antigen, CA125, CA19.9, p16, estrogen receptor, progesterone receptor, MUC6, PAX8, PAX2, p53, hepatocyte nuclear factor 1 beta, carbonic anhydrase IX, human epidermal receptor 2 (HER2), and mismatch repair (MMR) proteins. All markers were classified as negative, focal (<50% of tumor cells positive), or diffuse (≥50% tumor cells positive) except for p53 (classified as "wild-type" or "mutation-type"), HER2 (scored using the College of American Pathologists guidelines for gastric carcinomas), and MMR proteins (categorized as retained or lost). There was positive staining with CK7 (47/47-45 diffuse, 2 focal), MUC6 (17/21-6 diffuse, 11 focal), carcinoembryonic antigen (25/31-12 diffuse, 13 focal

  16. IMMUNOHISTOCHEMICAL AND BIOCHEMICAL ANALYSIS OF MAMMARY GLAND TUMOURS OF DIFFERENT AGE PATIENTS.

    PubMed

    Lykholat, T; Lykholat, O; Antonyuk, S

    2016-01-01

    Immunohistochemical and biochemical study of infiltrative ductal breast carcinoma and tissue adjacent to the tumour revealed a particular molecular profile and characteristics of the oxidant-antioxidant status neoplasms depending on the age of the patients and the presence of metastases in regional lymph nodes. Some causes of high aggressiveness and low hormone sensitivity of tumours in premenopausal women, as well as stability and high metastatic potential of tumours in postmenopausal women have been found. PMID:27266184

  17. Immunohistochemical evaluation of stem cell markers and signal transducer and activator of transcription 6 (STAT6) in solitary fibrous tumors.

    PubMed

    Wang, Chengyan; Qi, Yan; Liu, Ruixue; Lan, Jiaojiao; Zhou, Yang; Ju, Xinxin; Chen, Dongdong; Zou, Hong; Li, Shugang; Hu, Jianming; Zhao, Jin; Shen, Yaoyuan; Sun, Zhenzhu; Pang, Lijuan; Li, Feng

    2015-01-01

    Solitary fibrous tumors (SFT) are fibroblastic, ubiquitous mesenchymal tumors. Although several SFT studies have been conducted, the cell of origin of SFT remains controversial and reliable diagnostic markers are needed for SFT identification for proper prognosis and therapeutics. To analyze the immunophenotype of SFT for the identification of specific diagnostic markers and the cell of origin of this tumor, we performed an immunohistochemical study of stem cell markers [aldehyde dehydrogenase 1 (ALDH1), CD29, CD44, CD133, and nestin] and signal transducer and activator of transcription 6 (STAT6) in 18 cases of SFT. The results demonstrated that ALDH1 was present in 16 cases (16/18), STAT6 in 13 cases (13/18), CD44 in 8 cases (8/18), and CD29 in 1 case (1/18), whereas CD133 and nestin were absent in all cases (0/18). Our results indicate that combination with ALDH1 and STAT6 can improve the diagnostic value of CD34 for SFT. The immunohistochemical findings for stem cell surface markers indicate that SFT may originate from stem cells and that ALDH1 plays an important role in the development of SFT. PMID:26617768

  18. Immunohistochemical evaluation of stem cell markers and signal transducer and activator of transcription 6 (STAT6) in solitary fibrous tumors.

    PubMed

    Wang, Chengyan; Qi, Yan; Liu, Ruixue; Lan, Jiaojiao; Zhou, Yang; Ju, Xinxin; Chen, Dongdong; Zou, Hong; Li, Shugang; Hu, Jianming; Zhao, Jin; Shen, Yaoyuan; Sun, Zhenzhu; Pang, Lijuan; Li, Feng

    2015-01-01

    Solitary fibrous tumors (SFT) are fibroblastic, ubiquitous mesenchymal tumors. Although several SFT studies have been conducted, the cell of origin of SFT remains controversial and reliable diagnostic markers are needed for SFT identification for proper prognosis and therapeutics. To analyze the immunophenotype of SFT for the identification of specific diagnostic markers and the cell of origin of this tumor, we performed an immunohistochemical study of stem cell markers [aldehyde dehydrogenase 1 (ALDH1), CD29, CD44, CD133, and nestin] and signal transducer and activator of transcription 6 (STAT6) in 18 cases of SFT. The results demonstrated that ALDH1 was present in 16 cases (16/18), STAT6 in 13 cases (13/18), CD44 in 8 cases (8/18), and CD29 in 1 case (1/18), whereas CD133 and nestin were absent in all cases (0/18). Our results indicate that combination with ALDH1 and STAT6 can improve the diagnostic value of CD34 for SFT. The immunohistochemical findings for stem cell surface markers indicate that SFT may originate from stem cells and that ALDH1 plays an important role in the development of SFT.

  19. Comparison of Myofibroblasts Between Solid/Multicystic Ameloblastoma and Unicystic Ameloblastoma: An Immunohistochemical Analysis

    PubMed Central

    Shenoy, Sadhana; Narayan, Thondikulam Venakataraman; Jayaram, Ranjita

    2016-01-01

    Introduction Microenvironment is crucial for the maintenance of cellular functions and tissue integrity suggesting that cancer-induced changes in the stroma may contribute to cancer invasion and its biological behaviour. One of the major constituent of the tumour stroma is myofibroblasts. Myofibroblasts are differentiated host fibroblasts that express α-Sma as cytoplasmic microfilaments. They are considered as one of the modified stromal component which in recent years have been thought to have a role in the invasion and aggressive behaviour of odontogenic tumours too. Aim To detect immunohistochemically the presence of myofibroblasts in solid/multicystic ameloblastoma and in unicystic ameloblastoma and to see if a relationship exists between the frequency and pattern of distribution of myofibroblasts and the behaviour of ameloblastomas. Materials and Methods Ten cases each of solid/multicystic ameloblastoma and unicystic ameloblastoma were stained immunohistochemically for vimentin, α-SMA and desmin. The frequency and pattern of distribution of myofibroblasts in the two study groups were analysed and then compared with clinical and radiographic features of pain and cortical perforation respectively. Results Immunohistochemical reaction for α-SMA (alpha Smooth Muscle Actin) showed positive cells in the stroma of both solid/multicystic and unicystic ameloblastomas. The mean number of myofibroblasts was more in unicystic ameloblastoma (UA) compared to Solid/Multicystic Ameloblastoma (SMA). Myofibroblasts expression was dense and arranged in the form of fascicles with indistinct cell borders in one case of follicular ameloblastoma, two cases of plexiform ameloblastoma and in a focal area of one case of type 1UA. In all other cases where the expression was noted, the myofibroblasts were spindle in shape with distinct cell boundaries. Conclusion The results of the study indicate that myofibroblasts alone may not play a role in the behaviour of ameloblastomas. This

  20. Basal Cell Adenoma-Clinicopathological, Immunohistochemical Analysis and Surgical Considerations of a Rare Salivary Gland Tumor with Review of Literature

    PubMed Central

    Bhagat Singh, AD; Majumdar, Swapan; Ghosh, Amal Kanti; Gandi, Lakshmi; Choudaha, Nidhi; Sharma, Ipsita; Pal, SP

    2015-01-01

    Introduction: Basal cell adenoma (BCA) of the salivary glands is a rare benign salivary gland tumour. Differentiation of BCA from varied entities involving maxillofacial area is mandatory. Aim: To analyze the clinicopathological, histopathologic features, immunohistochemcal analysis and surgical considerations of this rare entity. Materials and Methods: This study included 12 cases of BCA from archives of department reported over the period of 13 years. All the pertaining clinicopathologic features such as incidence, age, sex and site of lesions were assessed. Tissue sections were stained by using panel of immunohistochemical markers, i.e. Pan CK, CK 5/6 and S100, Calponin, p63, CD 117 and smooth muscle actin. Results: BCA was observed in 26-52 years age group (mean age, 38.75 years) with female propensity of 7:5 male to female ratio. It is seen more commonly in parotid gland, followed by upper lip, buccal mucosa and palate. Solid type is the most common histopathologic type followed by tubular, membranous and trabecular. Only one case of membranous type of BCA showed recurrence. Pan CK, CK 5/6 showed strong immunoreactivity, calponin showed moderate staining, p63 and Ki-67 mild staining, whereas CD 117 and SMA showed negative immunostaining. Conclusion: Vigilant comprehensive analysis of all the pertaining clinicopathologic and histopathologic features and immunohistochemical analysis are required for differentiating from other lesions with basaloid differentiation having varying prognosis. PMID:25838763

  1. Complete chromogen separation and analysis in double immunohistochemical stains using Photoshop-based image analysis.

    PubMed

    Lehr, H A; van der Loos, C M; Teeling, P; Gown, A M

    1999-01-01

    Simultaneous detection of two different antigens on paraffin-embedded and frozen tissues can be accomplished by double immunohistochemistry. However, many double chromogen systems suffer from signal overlap, precluding definite signal quantification. To separate and quantitatively analyze the different chromogens, we imported images into a Macintosh computer using a CCD camera attached to a diagnostic microscope and used Photoshop software for the recognition, selection, and separation of colors. We show here that Photoshop-based image analysis allows complete separation of chromogens not only on the basis of their RGB spectral characteristics, but also on the basis of information concerning saturation, hue, and luminosity intrinsic to the digitized images. We demonstrate that Photoshop-based image analysis provides superior results compared to color separation using bandpass filters. Quantification of the individual chromogens is then provided by Photoshop using the Histogram command, which supplies information on the luminosity (corresponding to gray levels of black-and-white images) and on the number of pixels as a measure of spatial distribution. (J Histochem Cytochem 47:119-125, 1999)

  2. Immunohistochemical analysis of the Wnt/β-catenin signaling pathway in pancreatic neuroendocrine neoplasms

    PubMed Central

    Weiss, Vivian; Dueber, Julie; Wright, Jesse P; Cates, Justin; Revetta, Frank; Parikh, Alexander A; Merchant, Nipun B; Shi, Chanjuan

    2016-01-01

    AIM To investigate the role of the Wnt/β-catenin pathway in pancreatic neuroendocrine neoplasms (PanNENs). METHODS Tissue microarrays containing 88 PanNENs were immunohistochemically labeled with antibodies to β-catenin, E-cadherin, adenomatous polyposis coli (APC), chromogranin and synaptophysin. One case had only metastatic tumors resected, whereas others (n = 87) received pancreatectomy with or without partial hepatectomy. Pathology slides, demographic, clinicopathologic, and follow up data were reviewed. Patients’ demographics, clinicopathologic features, and immunohistochemical results from 87 primary tumors were compared between patients with low stage (stage I/II) and high stage (stage III/IV) tumors. In addition, correlation of immunohistochemical results from primary tumors with disease-specific survival (DSS) was evaluated. RESULTS Strong membranous β-catenin staining in the primary tumor was observed in all 13 stage III/IV PanNENs as compared to 47% (35/74) of stage I/II tumors (P < 0.01). However, the strong membranous β-catenin staining was unassociated with tumor grade or DSS. Decreased membranous β-catenin staining was associated with decreased membranous E-cadherin labeling. Nuclear β-catenin staining was seen in 15% (2/13) of stage III/IV PanNENs as compared to 0% (0/74) of stage I/II tumors (P = 0.02). The case with metastasectomy only also showed nuclear β-catenin staining. Two of the three cases with nuclear β-catenin staining were familial adenomatous polyposis (FAP) patients. Lack of APC expression was seen in 70% (57/81) of the cases, including the 3 cases with nuclear β-catenin staining. Expression of E-cadherin and APC in primary tumor was not correlated with tumor grade, tumor stage, or disease specific survival. CONCLUSION The Wnt/β-catenin pathway was altered in some PanNENs, but did not Impact DSS. PanNENs in FAP patients demonstrated nuclear β-catenin accumulation and loss of APC. PMID:27574554

  3. Activation analysis

    SciTech Connect

    Alfassi, Z.B. . Dept. of Nuclear Engineering)

    1990-01-01

    This volume contains 16 chapters on the application of activation analysis in the fields of life sciences, biological materials, coal and its effluents, environmental samples, archaeology, material science, and forensics. Each chapter is processed separately for the data base.

  4. Pathological, immunohistochemical and microbiologicalal analysis of lacrimal sac biopsies in patients with chronic dacrocystitis

    PubMed Central

    Amin, Rowayda Mahmoud; Hussein, Faten Aly; Idriss, Hisham Farouk; Hanafy, Nesrine Fathy; Abdallah, Dina Mohamed

    2013-01-01

    AIM To analyze cases of obstruction of the nasolacrimal duct which creates a fertile environment for secondary bacterial infection and can result in dacryocystitis, which is a constant threat to cornea and orbital soft tissue and a potential source of endophthalmitis following intraocular surgery. The majority of obstructions of the lacrimal excretory outflow system are acquired ones occurring in adulthood and involving the distal parts of the system. Acquired obstruction may be primary/idiopathic or secondary to a wide variety of infectious, inflammatory, traumatic, mechanical, toxic or neoplastic causes mimicking idiopathic inflammation. These cases are treated by dacryocystorhinostomy (DCR). METHODS The present study was conducted to determine the histopathologic, immunohistochemical and current microbiologic characteristics of lacrimal sac specimens in patients undergoing external dacryocystorhinostomy. RESULTS Non-specific lacrimal sac pathology was present in all 33 cases and 81.8% of the cases showed moderate chronic inflammation with a chronic inflammatory score (CIS) ranging between 4 and 6, whereas 12.12% showed severe inflammatory changes with a CIS of 7. Mild degree of inflammation was seen in 6.06% with a CIS of 3. The total prevalence of gram-positive, gram-negative, and culture-negative samples were 59.4%, 37.5%, and 3% respectively. CONCLUSION Non-specific chronic inflammation with fibrosis is indeed the most commonly reported histopathological finding in lacrimal sac wall biopsy specimens. PMID:24392331

  5. Ectopic primary intrathyroidal thymoma: a clinicopathological and immunohistochemical analysis of 3 cases.

    PubMed

    Weissferdt, Annikka; Moran, Cesar A

    2016-03-01

    Thymomas are rare tumors that occasionally arise from ectopic locations. Ectopic thymomas originating within the thyroid gland are an exceedingly uncommon clinical entity that has only been described sporadically. In this study, we present the clinicopathological and immunohistochemical features of 3 primary intrathyroidal thymomas. The patients were 2 women and 1 man between the ages of 43 and 53 years (average, 48 years). Clinically, the patients presented with neck pain or enlarged thyroid glands. Physical examination and thyroid ultrasound revealed the presence of nodular masses confined to the thyroid parenchyma. No concurrent mediastinal tumors were identified in any of the cases, and none of the patients had a history of thymoma. Fine needle aspirate performed in 1 case was interpreted as possibly Hashimoto thyroiditis. Surgical resection was performed in all cases. Grossly, the lesions were circumscribed masses measuring from 1 to 4 cm in size. Histologically, the lesions showed the classic biphasic cellular proliferation of thymomas characterized by varying proportions of epithelial cells and lymphocytes. Two patients remain alive and well 1.5 to 2 years after their surgical resection, whereas the third patient was lost to follow-up. The cases herein presented highlight an unusual tumor entity that can be clinically confused for more common lesions affecting the thyroid gland. Awareness of this entity is important to avoid misdiagnosis and secure appropriate clinical management. PMID:26826412

  6. Ultrastructural and immunohistochemical analysis of the 8-20 week human fetal pancreas.

    PubMed

    Riopel, Matthew; Li, Jinming; Fellows, George F; Goodyer, Cynthia G; Wang, Rennian

    2014-01-01

    Development of the human pancreas is well-known to involve tightly controlled differentiation of pancreatic precursors to mature cells that express endocrine- or exocrine-specific protein products. However, details of human pancreatic development at the ultrastructural level are limited. The present study analyzed 8-20 week fetal age human pancreata using scanning and transmission electron microscopy (TEM), TEM immunogold and double or triple immunofluorescence staining. Primary organization of islets and acini occurred during the developmental period examined. Differentiating endocrine and exocrine cells developed from the ductal tubules and subsequently formed isolated small clusters. Extracellular matrix fibers and proteins accumulated around newly differentiated cells during their migration and cluster formation. Glycogen expression was robust in ductal cells of the pancreas from 8-15 weeks of fetal age; however, this became markedly reduced at 20 weeks, with a concomitant increase in acinar cell glycogen content. Insulin secretory granules transformed from being dense and round at 8 weeks to distinct geometric (multilobular, crystalline) structures by 14-20 weeks. Initially many of the differentiating endocrine cells were multihormonal and contained polyhormonal granules; by 20 weeks, monohormonal cells were in the majority. Interestingly, certain secretory granules in the early human fetal pancreatic cells showed positivity for both exocrine (amylase) and endocrine proteins. This combined ultrastructural and immunohistochemical study showed that, during early developmental stages, the human pancreas contains differentiating epithelial cells that associate closely with the extracellular matrix, have dynamic glycogen expression patterns and contain polyhormonal as well as mixed endocrine/exocrine granules.

  7. Calcium hydroxylapatite filler for facial rejuvenation: a histologic and immunohistochemical analysis.

    PubMed

    Berlin, Alexander L; Hussain, Mussarratt; Goldberg, David J

    2008-06-01

    BACKGROUND Soft-tissue augmentation using calcium hydroxylapatite (CaHA) filler has been shown to be a safe and effective filler agent. A previous study utilizing electron microscopy demonstrated deposition of collagen around filler microspheres with minimal inflammatory response. OBJECTIVE The purpose of this study was to further characterize the expression of collagen in response to injected CaHA filler material using conventional, special, and immunohistochemical (IHC) staining. MATERIALS AND METHODS Five subjects with mild to moderate nasolabial rhytids underwent a single treatment with CaHA filler. Additionally, 0.1 to 0.2 mL of the filler material was injected subdermally into the postauricular area. Six months following the treatment, all subjects underwent biopsy of the treated postauricular area. Biopsies were analyzed using hematoxylin and eosin staining, special staining with picrosirius red (PSR), and IHC staining for collagen Types I and III. RESULTS All subjects completed the study with no complications. Biopsy specimens revealed increased deposition of collagen around the filler material, which was confirmed and further characterized using PSR and IHC staining. CONCLUSION Soft tissue augmentation with CaHA filler leads to long-term deposition of new collagen surrounding filler microspheres, which may contribute to the overall improvement in the appearance of treated rhytids.

  8. Immunohistochemical and molecular analysis of caveolin-1 expression in canine mammary tumors.

    PubMed

    Zuccari, D A P C; Castro, R; Gavioli, A F; Mancini, U M; Frade, C S; Leonel, C

    2012-01-01

    Caveolin-1 (Cav-1) is a structural protein present in invaginations of the cell membrane. In human breast cancer, the cav-1 gene is believed to be a tumor suppressor gene associated with inhibition of tumor metastasis. However, little is known about its expression, regulation and function in canine mammary tumors. Expression levels of cav-1 were investigated using real-time PCR and immunohistochemical detection with an anti-human Cav-1 antibody. Gene expression stability of different samples was analyzed using the geNorm software. Mammary tumors from 51 female dogs were compared to normal mammary tissue from 10 female dogs. Malignant mammary cells showed a loss of Cav-1 expression by quantitative RT-PCR and weak Cav-1 staining by immunohistochemistry compared to normal mammary gland tissue. There was a significant relationship between outcome and immunostaining as well as with tumor size, indicating that caveolin subexpression has a positive predictive value and is related to higher survival and smaller tumor size. Our findings indicate that Cav-1 is a potential prognostic marker for canine mammary tumors. PMID:22370882

  9. Immunohistochemical Analysis of Paraoxonases and Chemokines in Arteries of Patients with Peripheral Artery Disease

    PubMed Central

    Hernández-Aguilera, Anna; Sepúlveda, Julio; Rodríguez-Gallego, Esther; Guirro, Maria; García-Heredia, Anabel; Cabré, Noemí; Luciano-Mateo, Fedra; Fort-Gallifa, Isabel; Martín-Paredero, Vicente; Joven, Jorge; Camps, Jordi

    2015-01-01

    Oxidative damage to lipids and lipoproteins is implicated in the development of atherosclerotic vascular diseases, including peripheral artery disease (PAD). The paraoxonases (PON) are a group of antioxidant enzymes, termed PON1, PON2, and PON3 that protect lipoproteins and cells from peroxidation and, as such, may be involved in protection against the atherosclerosis process. PON1 inhibits the production of chemokine (C–C motif) ligand 2 (CCL2) in endothelial cells incubated with oxidized lipoproteins. PON1 and CCL2 are ubiquitously distributed in tissues, and this suggests a joint localization and combined systemic effect. The aim of the present study has been to analyze the quantitative immunohistochemical localization of PON1, PON3, CCL2 and CCL2 receptors in a series of patients with severe PAD. Portions of femoral and/or popliteal arteries from 66 patients with PAD were obtained during surgical procedures for infra-inguinal limb revascularization. We used eight normal arteries from donors as controls. PON1 and PON3, CCL2 and the chemokine-binding protein 2, and Duffy antigen/chemokine receptor, were increased in PAD patients. There were no significant changes in C–C chemokine receptor type 2. Our findings suggest that paraoxonases and chemokines play an important role in the development and progression of atherosclerosis in peripheral artery disease. PMID:25993297

  10. Gingival juvenile xanthogranuloma in an adult patient: case report with immunohistochemical analysis and literature review.

    PubMed

    Consolaro, Alberto; Sant'Ana, Eduardo; Lawall, Melaine A; Consolaro, Maria Fernanda M O; Bacchi, Carlos Eduardo

    2009-02-01

    Juvenile xanthogranuloma (JXG) is a non-Langerhans cell histiocytosis (nonLCH). It is a benign and self-healing disorder that generally affects infants and children. Oral lesions in adult patients are rare, although the microscopic findings are similar to those observed in other locations. A 56-year-old white man presented with a chief complaint of a gingival mass that had appeared 6 months before and had grown slowly. An intraoral examination revealed the presence of a solitary, softened gingival mass affecting the mandibular lingual gingiva at the right central incisor area. A biopsy of the lesion showed multiple large macrophages and numerous giant cells of Touton type. The immunohistochemistry positivity for CD68, fascin, factor XIIIa, alpha-antitrypsin and negativity for S-100, beta-actin, CD1a, and desmin confirmed the diagnosis of JXG. The occurrence of adult oral JXG is extremely rare. It is a nonLCH that may present variable clinical and microscopic aspects, which leads to a diversity of clinical misdiagnoses. A precise diagnosis of these lesions requires an accurate evaluation of clinical, microscopic, and immunohistochemical features.

  11. Immunohistochemical and ultrastructural analysis of medullary carcinoma of the thyroid in relation to hormone production.

    PubMed Central

    Kameya, T.; Shimosato, Y.; Adachi, I.; Abe, K.; Kasai, N.; Kimura, K.; Baba, K.

    1977-01-01

    Eighteen cases of medullary carcinoma of the thyroid were analyzed immunohistochemically for calcitonin- and ACTH-containing cells, and tumor cells in 8 cases were examined by an electron microscope and analyzed by manual and computer procedures with particular attention paid to the size and quality of secretory granules. Calcitonin- and ACTH-containing cells were found singly or in clusters in 14 and 11 tumors, respectively. In 4 cases, calcitonin-positive cell clusters and an increase in number of singly scattered C cells were seen apart from the main tumor, suggesting a multicentric nature of certain medullary carcinomas. Some ACTH-containing cells were apparently also positive for calcitonin. In a case of familial Sipple disease, follicular lining cells were replaced in areas with ACTH-containing cells. Three to five frequency distribution curves of the size of secretory granules were obtained in all of 6 cases analyzed, and at least two different types of granule matrix were identified. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12 Figure 13 Figure 14 Figure 15 Figure 16 Figure 17 Figure 18 Figure 19 PMID:202164

  12. Immunohistochemical analysis of the hypothalamic-pituitary-adrenal axis in dogs: Sex-linked and seasonal variation.

    PubMed

    Gallelli, M F; Lombardo, D; Vissio, P; Quiroga, A; Caggiano, N; Soler, E; Meikle, A; Castillo, V A

    2016-02-01

    This study evaluated sexual dimorphism and seasonal variations in corticotrophs and adrenal zona fasciculata in dogs, as well as the expression of oestrogen receptor alpha (ERα). An immunohistochemical analysis was conducted in pituitaries for ACTH and in adrenal glands for ERα and for the melanocortin-2-receptor (MC2R) in winter and summer. Double immunofluorescence was performed to identify ERα in corticotrophs. Females had a greater proportion of corticotrophs per field (p<0.01), with a greater cellular area and optical density (p<0.001) than males. Optical density of corticotrophs was greater in winter for both sexes (p<0.001). In zona fasciculata, ERα and MC2R expression was greater in females (p<0.001) and was greater in winter (p<0.001). ERα was identified in corticotrophs. This study is the first to demonstrate ERα expression in corticotrophs and the adrenal cortex in dogs, providing evidence for sexual dimorphism and seasonal variations. PMID:26850531

  13. Immunohistochemical analysis of the hypothalamic-pituitary-adrenal axis in dogs: Sex-linked and seasonal variation.

    PubMed

    Gallelli, M F; Lombardo, D; Vissio, P; Quiroga, A; Caggiano, N; Soler, E; Meikle, A; Castillo, V A

    2016-02-01

    This study evaluated sexual dimorphism and seasonal variations in corticotrophs and adrenal zona fasciculata in dogs, as well as the expression of oestrogen receptor alpha (ERα). An immunohistochemical analysis was conducted in pituitaries for ACTH and in adrenal glands for ERα and for the melanocortin-2-receptor (MC2R) in winter and summer. Double immunofluorescence was performed to identify ERα in corticotrophs. Females had a greater proportion of corticotrophs per field (p<0.01), with a greater cellular area and optical density (p<0.001) than males. Optical density of corticotrophs was greater in winter for both sexes (p<0.001). In zona fasciculata, ERα and MC2R expression was greater in females (p<0.001) and was greater in winter (p<0.001). ERα was identified in corticotrophs. This study is the first to demonstrate ERα expression in corticotrophs and the adrenal cortex in dogs, providing evidence for sexual dimorphism and seasonal variations.

  14. A New Cell Block Method for Multiple Immunohistochemical Analysis of Circulating Tumor Cells in Patients with Liver Cancer

    PubMed Central

    Nam, Soo Jeong; Yeo, Hyun Yang; Chang, Hee Jin; Kim, Bo Hyun; Hong, Eun Kyung; Park, Joong-Won

    2016-01-01

    Purpose We developed a new method of detecting circulating tumor cells (CTCs) in liver cancer patients by constructing cell blocks from peripheral blood cells, including CTCs, followed by multiple immunohistochemical analysis. Materials and Methods Cell blockswere constructed from the nucleated cell pellets of peripheral blood afterremoval of red blood cells. The blood cell blocks were obtained from 29 patients with liver cancer, and from healthy donor blood spikedwith seven cell lines. The cell blocks and corresponding tumor tissues were immunostained with antibodies to seven markers: cytokeratin (CK), epithelial cell adhesion molecule (EpCAM), epithelial membrane antigen (EMA), CK18, α-fetoprotein (AFP), Glypican 3, and HepPar1. Results The average recovery rate of spiked SW620 cells from blood cell blocks was 91%. CTCs were detected in 14 out of 29 patients (48.3%); 11/23 hepatocellular carcinomas (HCC), 1/2 cholangiocarcinomas (CC), 1/1 combined HCC-CC, and 1/3 metastatic cancers. CTCs from 14 patients were positive for EpCAM (57.1%), EMA (42.9%), AFP (21.4%), CK18 (14.3%), Gypican3 and CK (7.1%, each), and HepPar1 (0%). Patients with HCC expressed EpCAM, EMA, CK18, and AFP in tissue and/or CTCs, whereas CK, HepPar1, and Glypican3 were expressed only in tissue. Only EMA was significantly associated with the expressions in CTC and tissue. CTC detection was associated with higher T stage and portal vein invasion in HCC patients. Conclusion This cell block method allows cytologic detection and multiple immunohistochemical analysis of CTCs. Our results show that tissue biomarkers of HCC may not be useful for the detection of CTC. EpCAM could be a candidate marker for CTCs in patients with HCC. PMID:27034142

  15. Systematic review and meta-analysis of immunohistochemical prognostic biomarkers in resected oesophageal adenocarcinoma

    PubMed Central

    McCormick Matthews, L H; Noble, F; Tod, J; Jaynes, E; Harris, S; Primrose, J N; Ottensmeier, C; Thomas, G J; Underwood, T J

    2015-01-01

    Background: Oesophageal adenocarcinoma (OAC) is one of the fastest rising malignancies with continued poor prognosis. Many studies have proposed novel biomarkers but, to date, no immunohistochemical markers of survival after oesophageal resection have entered clinical practice. Here, we systematically review and meta-analyse the published literature, to identify potential biomarkers. Methods: Relevant articles were identified via Ovid medline 1946–2013. For inclusion, studies had to conform to REporting recommendations for tumor MARKer (REMARK) prognostic study criteria. The primary end-point was a pooled hazard ratio (HR) and variance, summarising the effect of marker expression on prognosis. Results: A total of 3059 articles were identified. After exclusion of irrelevant titles and abstracts, 214 articles were reviewed in full. Nine molecules had been examined in more than one study (CD3, CD8, COX-2, EGFR, HER2, Ki67, LgR5, p53 and VEGF) and were meta-analysed. Markers with largest survival effects were COX-2 (HR=2.47, confidence interval (CI)=1.15–3.79), CD3 (HR=0.51, 95% CI=0.32–0.70), CD8 (HR=0.55, CI=0.31–0.80) and EGFR (HR=1.65, 95% CI=1.14–2.16). Discussion: Current methods have not delivered clinically useful molecular prognostic biomarkers in OAC. We have highlighted the paucity of good-quality robust studies in this field. A genome-to-protein approach would be better suited for the development and subsequent validation of biomarkers. Large collaborative projects with standardised methodology will be required to generate clinically useful biomarkers. PMID:26110972

  16. A detailed appraisal of mesocolic lymphangiology – an immunohistochemical and stereological analysis

    PubMed Central

    Culligan, Kevin; Sehgal, Rishabh; Mulligan, Daniel; Dunne, Colum; Walsh, Stewart; Quondamatteo, Fabio; Dockery, Peter; Coffey, J Calvin

    2014-01-01

    Inadequate resection of the adjoining mesentery is associated with adverse outcome for colon cancer. Disruption of the integrity of the mesenteric lymphatic package has been implicated in this, though not proven. Recent studies have determined mesenteric anatomy and histology and now provide an opportunity to determine accurately the distribution of lymphatic vessels. The aim of this study was to characterise the distribution of the lymphatic vessels (LV) within the small intestinal and colonic mesentery, and in Toldt's fascia, which lies between the mesocolon and underlying retroperitoneum. Mesenteric samples were harvested from 12 human cadavers. Samples were taken from the small bowel mesentery, ascending, transverse, descending mesocolon and from both apposed and non-apposed portions of the mesosigmoid. Serial sections were stained immunohistochemically with monoclonal antibody D2-40 (podoplanin), and Masson's Trichrome. Lymphatic vessel (LV) density and radius of diffusion were determined using a stereological approach. A lymphatic network was embedded within the mesenteric connective tissue lattice throughout each mesenteric region. LV were identifiable within the submesothelial connective tissue where they measured 10.2 ± 4.1 μm in diameter and had an average radius of diffusion of 174.72 ± 97.68 μm. Unexpectedly, LV were identified in Toldt's fascia, where they measured 4.3 ± 3.1 μm in diameter and had a radius of diffusion of 165.12 ± 66.26 μm. This is the first study systematically to determine and quantify the distribution of lymphatic vessels within the mesenteric organ and to demonstrate the presence of such vessels within Toldt's fascia. A rich lymphatic network occupies all levels of the mesenteric connective tissue lattice. Within the latter, they are found within 0.1 mm of peritonealised mesenteric surfaces and are separated by an average distance of 0.17 mm and may be particularly vulnerable during surgery. PMID:25040735

  17. Ultrastructural, immunohistochemical and biochemical analysis of glycosaminoglycans and proteoglycans in the mouse pubic symphysis during pregnancy.

    PubMed

    Pinheiro, M C; Mora, O A; Caldini, E G; Battlehner, C N; Joazeiro, P P; Toledo, O M S

    2005-06-01

    During pregnancy, an interpubic ligament is formed in the mouse pubic symphysis. In late stages, this ligament undergoes "relaxation" to allow proper delivery, which is expected on the 19th day. Proteoglycans and hyaluronic acid play an important role in the remodeling of the extracellular matrix in these tissues. Glycosaminoglycans and proteoglycans were studied by electron microscopic, immunohistochemical and biochemical methods in samples of mouse pubic symphysis from the 12th to 18th day of pregnancy. At the ultrastructural level, using cuprolinic blue and enzymatic digestion by chondroitin lyases, two types of proteoglycan filaments were observed in the fibrocartilage on the 12th day, as well as in D 15, D 17 and D 18 pubic ligaments. The only sulfated glycosaminoglycan in these filaments was chondroitin sulfate, as shown by chondroitin lyase treatment. Their electrophoretic mobility, before and after enzymatic degradation, corroborated this inference. The ratio of chondroitin sulfate/dry weight of symphysis showed two phases of increase: between D12 and D 15, and between D 17 and D 18. We suggest that the first corresponds mainly to an increase in decorin when the ligament is formed, and the second to versican, during "relaxation". Versican and hyaluronic acid, working as water holding molecules would be responsible for the hydration of the ligament at the end of pregnancy, allowing an increase in resiliency. The presence of hyaluronic acid was confirmed by labeling with HA-probe in the perichondrium, fibrocartilage and ligament. The role of collagen fibers as physical restrictors of the complete expansion of glycosaminoglycans and hyaluronic acid in tissue is discussed. PMID:15951206

  18. Immunohistochemical quantitation of oestrogen receptors and proliferative activity in oestrogen receptor positive breast cancer.

    PubMed Central

    Jensen, V; Ladekarl, M

    1995-01-01

    AIM--To evaluate the effect of the duration of formalin fixation and of tumour heterogeneity on quantitative estimates of oestrogen receptor content (oestrogen receptor index) and proliferative activity (MIB-1 index) in breast cancer. METHODS--Two monoclonal antibodies, MIB-1 and oestrogen receptor, were applied to formalin fixed, paraffin wax embedded tissue from 25 prospectively collected oestrogen receptor positive breast carcinomas, using a microwave antigen retrieval method. Tumour tissue was allocated systematically to different periods of fixation to ensure minimal intraspecimen variation. The percentages of MIB-1 positive and oestrogen receptor positive nuclei were estimated in fields of vision sampled systematically from the entire specimen and from the whole tumour area of one "representative" cross-section. RESULTS--No correlation was found between the oestrogen receptor and MIB-1 indices and the duration of formalin fixation. The estimated MIB-1 and oestrogen receptor indices in tissue sampled systematically from the entire tumour were closely correlated with estimates obtained in a "representative" section. The intra- and interobserver correlation of the MIB-1 index was good, although a slight systematical error at the second assessment of the intraobserver study was noted. CONCLUSION--Quantitative estimates of oestrogen receptor content and proliferative activity are not significantly influenced by the period of fixation in formalin, varying from less than four hours to more than 48 hours. The MIB-1 and the oestrogen receptor indices obtained in a "representative" section do not deviate significantly from average indices determined in tissue samples from the entire tumour. Finally, the estimation of MIB-1 index is reproducible, justifying its routine use. PMID:7629289

  19. Host inflammatory response to polypropylene implants: insights from a quantitative immunohistochemical and birefringence analysis in a rat subcutaneous model

    PubMed Central

    Prudente, Alessandro; Fávaro, Wágner José; Latuf, Paulo; Riccetto, Cássio Luis Zanettini

    2016-01-01

    ABSTRACT Objectives To describe acute and sub acute aspects of histological and immunohistochemical response to PP implant in a rat subcutaneous model based on objective methods. Materials and Methods Thirty rats had a PP mesh subcutaneously implanted and the same dissection on the other side of abdomen but without mesh (sham). The animals were euthanized after 4 and 30 days. Six slides were prepared using the tissue removed: one stained with hematoxylin-eosin (inflammation assessment); one unstained (birefringence evaluation) and four slides for immunohistochemical processing: IL-1 and TNF-α (pro-inflammatory cytokines), MMP-2 (collagen metabolism) and CD-31 (angiogenesis). The area of inflammation, the birefringence index, the area of immunoreactivity and the number of vessels were objectively measured. Results A larger area of inflammatory reaction was observed in PP compared to sham on the 4th and on the 30th day (p=0.0002). After 4 days, PP presented higher TNF (p=0.0001) immunoreactivity than sham and no differences were observed in MMP-2 (p=0.06) and IL-1 (p=0.08). After 30 days, a reduction of IL-1 (p=0.010) and TNF (p=0.016) for PP and of IL-1 (p=0.010) for sham were observed. Moreover, area of MMP-2 immunoreactivity decreased over time for PP group (p=0.018). Birefringence index and vessel counting showed no differences between PP and sham (p=0.27 and p=0.58, respectively). Conclusions The implantation of monofilament and macroporous polypropylene in the subcutaneous of rats resulted in increased inflammatory activity and higher TNF production in the early post implant phase. After 30 days, PP has similar cytokines immunoreactivity, vessel density and extracellular matrix organization. PMID:27286125

  20. Immunohistochemical Analysis of Scarring Trachoma Indicates Infiltration by Natural Killer and Undefined CD45 Negative Cells

    PubMed Central

    Hu, Victor H.; Luthert, Philip J.; Pullin, James; Weiss, Helen A.; Massae, Patrick; Mtuy, Tara; Makupa, William; Essex, David; Mabey, David C. W.; Bailey, Robin L.; Holland, Martin J.; Burton, Matthew J.

    2016-01-01

    Introduction The phenotype and function of immune cells infiltrating the conjunctiva in scarring trachoma have yet to be fully characterized. We assessed tissue morphology and immunophenotype of cellular infiltrates found in trachomatous scarring compared to control participants. Methodology Clinical assessments and conjunctival biopsy samples were obtained from 34 individuals with trachomatous scarring undergoing trichiasis surgery and 33 control subjects undergoing cataract or retinal detachment surgery. Biopsy samples were fixed in buffered formalin and embedded in paraffin wax. Hematoxylin and eosin (H&E) staining was performed for assessment of the inflammatory cell infiltrate. Immunohistochemical staining of single markers on individual sections was performed to identify cells expressing CD3 (T-cells), CD4 (helper T-cells), CD8 (suppressor/cytotoxic T-cells and Natural Killer, NK, cells), NCR1 (NK cells), CD20 (B-cells), CD45 (nucleated hematopoietic cells), CD56 (NK and T-cells), CD68 (macrophages/monocytes) and CD83 (mature dendritic cells). The degree of scarring was assessed histologically using cross-polarized light to visualize collagen fibres. Principle Findings Scarring, regardless of clinical inflammation, was associated with increased inflammatory cell infiltrates on H&E and CD45 staining. Scarring was also associated with increased CD8+ and CD56+ cells, but not CD3+ cells, suggestive of a NK cell infiltrate. This was supported by the presence of NCR1+ cells. There was some increase in CD20+ cells, but no evidence for increased CD4+, CD68+ or CD83+ cells. Numerous CD45 negative cells were also seen in the population of infiltrating inflammatory cells in scarred conjunctiva. Disorganization of the normal collagen architecture was strongly associated with clinical scarring. Conclusions/Significance These data point to the infiltration of immune cells with a phenotype suggestive of NK cells in conjunctival trachomatous scarring. A large proportion of

  1. Expression of Proteinase-activated Receptor-2 in the Esophageal Mucosa of Gastroesophageal Reflux Disease Patients: A Histomorphologic and Immunohistochemical Study.

    PubMed

    Abd El-Rehim, Dalia M; Fath El-Bab, Hanaa K; Kamal, Enas M

    2015-10-01

    Data are limited regarding the role of proteinase-activated receptor-2 (PAR-2) in the esophageal mucosa in gastroesophageal reflux disease (GERD) patients. Our aim was to study PAR-2 expression and its relationship with different GERD-related clinical and pathologic parameters. Histomorphologic alterations in eosophageal mucosa in nonerosive reflux disease (NERD) and erosive reflux disease (ERD) were also, evaluated. Endoscopic biopsies of the esophageal mucosa were obtained from 94 GERD patients and 20 participants for histopathologic analysis and PAR-2 immunohistochemical staining. The present study demonstrated significantly higher PAR-2 expression in GERD patients compared with control, whereas no significant differences were seen between NERD and ERD groups. PAR-2 expression significantly correlated with histologic score (r=0.572, P<0.001) and severity of heartburn (r=0.541, P<0.001). PAR-2 expression was significantly associated with basal cell hyperplasia, and dilated intercellular spaces and inflammatory cell count (P<0.05). Histologic analysis revealed GERD-related histomorphologic alterations in the esophageal mucosa of GERD patients with significant differences (P<0.05) among groups. Total histologic score was significantly correlated with heartburn (r=0.299, P=0.025) and endoscopic severity (r=0.359, P=0.027) in NERD and ERD patients, respectively. Taken together, this study provides evidence for the major role of PAR-2 in the pathogenesis of GERD and GERD-associated mucosal alterations.

  2. Molecular cloning, immunohistochemical localization, characterization and expression analysis of caspase-8 from the blunt snout bream (Megalobrama amblycephala) exposed to ammonia.

    PubMed

    Sun, Shengming; Ge, Xianping; Zhu, Jian; Zhang, Wuxiao; Zhang, Qiong

    2015-12-01

    Caspase-8 is an initiator caspase that plays a crucial role in some cases of apoptosis by extrinsic and intrinsic pathways. Caspase-8 structure and function have been extensively studied in mammals, but in fish the characterization of that initiator caspase is still scarce. In this study, we isolated the caspase-8 gene from Megalobrama amblycephala, one of the most important industrial aquatic animals in China using rapid amplification of cDNA ends (RACE). The 2034 bp full-length M. amblycephala caspase-8 cDNA sequence contained an ORF of 1467 bp encoding a polypeptide of 489 amino acid residues, a 5'-UTR of 102 bp and a 3'-UTR of 462 bp. The caspase-8 amino acid sequences contained two highly conservative death effector domains (DEDs) at N-terminal, the caspase family domains P20 and P10, caspase-8 active-site pentapeptide and potential aspartic acid cleavage sites. Phylogenetic analysis revealed that M. amblycephala caspase-8 were clustered with the caspase-8 from other vertebrate. Real-time quantitative PCR analysis revealed that caspase-8 transcripts were detected in liver after exposure to ammonia. Meanwhile using Western blot analysis, caspase-8 cleaved fragment was detected and significant alteration of procaspase-8 level was found with the same ammonia treatment condition. Furthermore, the result of immunohistochemical detection showed that remarkable changes of immunopositive staining were observed after ammonia treatment. Accordingly, the results signify that caspase-8 of fish may play an essential role in ammonia induced apoptosis.

  3. Noninvasive device readouts validated by immunohistochemical analysis enable objective quantitative assessment of acute wound healing in human skin.

    PubMed

    Ud-Din, Sara; Greaves, Nicholas S; Sebastian, Anil; Baguneid, Mohamed; Bayat, Ardeshir

    2015-01-01

    Objective evaluation of cutaneous wounds through use of noninvasive devices has important implications for diagnosis, monitoring treatment efficacy, progression and may lead to development of improved theranostic treatment strategies. However, there is a lack of validation in the use of certain devices in wound repair, where objective measurements taken by noninvasive devices have been corroborated by immunohistochemical analysis. Thus, data from three acute wound-healing studies in healthy volunteers using three noninvasive objective devices were further evaluated by immunohistochemistry. One hundred ten participants had 5-mm diameter skin biopsies to their arms. Spectrophotometric intracutaneous analysis (SIAscopy), full-field laser perfusion imaging, and three-dimensional imaging provided quantitative measurements of melanin, hemoglobin, collagen, blood flow, and wound size; all of which were validated by immunohistochemistry. Full-field laser perfusion imaging showed blood flow increased to D7 and decreased by 40% to D14. SIAscopy showed that hemoglobin increased to D7 and reduced to D14. CD31 analysis corroborated this by showing a 76% increase in blood vessel density to D7 and a reduction by 14% to D14. Three-dimensional imaging showed that wound surface area reduced by 50% from day 7 to day 14. Alpha-smooth muscle Actin (Alpha-SMA) staining supported these trends by showing increased levels by 72% from D0 to D14 (corresponding to wound contraction). Collagen, measured by SIAscopy, decreased to D7 and increased to D14, which was validated by collagen III analysis. Additionally, collagen I increased by 14% from D0 to D14. SIAscopy measurements for melanin showed an increase at D7 and a slight reduction to D14, while melanogenesis increased by 46.7% from D0 to D14. These findings show the utility of noninvasive objective devices in the quantitative evaluation of wound-healing parameters in human skin as corroborated by immunohistochemistry. This may contribute

  4. Immunohistochemical analysis of factors related to apoptosis and cellular proliferation in relation to inflammation in dentigerous and odontogenic keratocyst

    PubMed Central

    Sreedhar, Gadiputi; Raju, M. Vijaya; Metta, Kiran Kumar; Manjunath, SM; Shetty, Sujan; Agarwal, Rakesh Kumar

    2014-01-01

    Background: The effect of inflammation on pathogenesis and biological behavior of odontogenic keratocyst (OKC) and dentigerous cyst (DC) is not completely understood. Hence, we aimed to analyze the effect of inflammation on biological behavior of OKC and DC using a proliferative and anti-apoptotic marker, i.e., proliferative cellular nuclear antigen (PCNA) and Bcl-2, respectively. Materials and Methods: Immunohistochemical staining was performed using anti-PCNA and Bcl-2 antibody in 10 cases each of classical OKC, inflamed OKC and classical DC and inflamed DC. Results: Inflamed OKC and DC showed a significant increase in PCNA expression and decrease in Bcl-2 expression when compared with non-inflamed cyst. Correlation between inflammation and proliferative and anti-apoptotic activity was found to be statistically non-significant. Conclusion: Inflammation is responsible for change in behavior of neoplastic epithelium of OKC and hence should be treated meticulously, whereas in DC it is responsible for changes in the epithelial lining. PMID:24678208

  5. p53 gene alterations in prostate cancer after radiation failure and their association with clinical outcome: a molecular and immunohistochemical analysis.

    PubMed

    Rakozy, C; Grignon, D J; Li, Y; Gheiler, E; Gururajanna, B; Pontes, J E; Sakr, W; Wood, D P; Sarkar, F H

    1999-01-01

    This study evaluates the prevalence of p53 gene mutations in prostate cancer in salvage prostatectomies after radiation failure using single strand conformational polymorphism (SSCP) and direct sequencing of the polymerase chain reaction (PCR) product. Findings were correlated with immunohistochemically (IHC) detectable p53 expression in residual prostate cancer. The usefulness of p53 as a marker of clinical outcome was evaluated. Thirty-three cases were available for molecular and immunohistochemical analysis. Immunohistochemical stains for p53 were performed with clone DO7. PCR-SSCP for mutations in the coding region of p53 DNA (exons 4-9) was performed on all immunopositive cases and 12 of 23 immunonegative cases. All samples with an SSCP shift were sequenced for the respective exon. Patients were evaluated for biochemical failure for 1-82 months (median 38 months) following surgery. Immunohistochemical p53 reactivity was noted in 10 of 33 (30%) patients. Among p53 immunopositive cases SSCP shifts were seen in 7 of 10 (70%) samples with 5 of the 7 (71%) showing p53 mutations. Univariate analysis revealed abnormal expression of p53 protein by immunohistochemistry to be a significant predictor of poorer outcome (p = 0.025, log rank), however this was not independent of pathologic stage, surgical margin status and Gleason score. The presence of p53 gene mutations by PCR-SSCP and direct sequencing did not predict for outcome. In our study 30% of prostate cancers at the time of salvage prostatectomy after radiation failure expressed immunohistochemically detectable p53. PCR-SSCP and sequencing shows that not all of these cases have detectable mutations in the most frequent mutation sites (exons 4-9). Clinical failure is more common in the group of prostate cancer patients with abnormal p53 immunoreactivity.

  6. Postnatal Growth of the Human Pons: A Morphometric and Immunohistochemical Analysis

    PubMed Central

    Tate, Matthew C.; Lindquist, Robert A.; Nguyen, Thuhien; Sanai, Nader; Barkovich, A. James; Huang, Eric J.; Rowitch, David H.; Alvarez-Buylla, Arturo

    2014-01-01

    Despite its critical importance to global brain function, the postnatal development of the human pons remains poorly understood. In the present study, we first performed MRI-based morphometric analyses of the postnatal human pons (0–18 years; n=6–14/timepoint). Pons volume increased 6-fold from birth to 5 years, followed by continued slower growth throughout childhood. The observed growth was primarily due to expansion of the basis pontis. T2-based MRI analysis suggests that this growth is linked to increased myelination, and histological analysis of myelin basic protein in human postmortem specimens confirmed a dramatic increase in myelination during infancy. Analysis of cellular proliferation revealed many Ki67+ cells during the first 7 months of life, particularly during the first month where proliferation was increased in the basis relative to tegmentum. The majority of proliferative cells in the postnatal pons expressed the transcription factor Olig2, suggesting an oligodendrocyte lineage. The proportion of proliferating cells that were Olig2+ was similar through the first 7 months of life and between basis and tegmentum. The number of Ki67+ cells declined dramatically from birth to 7 months and further decreased by 3 years, with a small number of Ki67+ cells observed throughout childhood. In addition, two populations of vimentin/nestin-expressing cells were identified: a dorsal group near the ventricular surface, which persists throughout childhood, and a parenchymal population that diminishes by 7 months and was not evident later in childhood. Together, our data reveal remarkable postnatal growth in the ventral pons, particularly during infancy when cells are most proliferative and myelination increases. PMID:25307966

  7. Immunohistochemical analysis of RTKs expression identified HER3 as a prognostic indicator of gastric cancer.

    PubMed

    Ema, Akira; Yamashita, Keishi; Ushiku, Hideki; Kojo, Ken; Minatani, Naoko; Kikuchi, Mariko; Mieno, Hiroaki; Moriya, Hiromitsu; Hosoda, Kei; Katada, Natsuya; Kikuchi, Shiro; Watanabe, Masahiko

    2014-12-01

    Standard treatment in Japan for the 13th Japanese Gastric Cancer Association stage II/III advanced gastric cancer is postoperative adjuvant S-1 administration after curative surgery. High expression of receptor type tyrosine kinases (RTKs) has repeatedly represented poor prognosis for cancers. However it has not been demonstrated whether RTKs have prognostic relevance for stage II/III gastric cancer with standard treatment. Tumor tissues were obtained from 167 stage II/III advanced gastric cancer patients who underwent curative surgery and received postoperative S-1 chemotherapy from 2000 to 2010. Expression of the RTKs including EGFR, HER2, HER3, IGF-1R, and EphA2 was analyzed using immunohistochemistry (IHC). Analysis using a multivariate proportional hazard model identified the most significant RTKs that represented independent prognostic relevance. When tumor HER3 expression was classified into IHC 1+/2+ (n = 98) and IHC 0 (n = 69), the cumulative 5-year Relapse Free Survival (5y-RFS) was 56.5 and 82.9%, respectively (P = 0.0034). Significant prognostic relevance was similarly confirmed for IGF-1R (P = 0.014), and EGFR (P = 0.030), but not for EphA2 or HER2 expression. Intriguingly, HER3 expression was closely correlated with IGF-1R (P < 0.0001, R = 0.41), and EphA2 (P < 0.0001, R = 0.34) expression. Multivariate proportional hazard model analysis identified HER3 (IHC 1+/2+) (HR; 1.53, 95% CI, 1.11-2.16, P = 0.0078) as the sole RTK that was a poor prognostic factor independent of stage. Of the 53 patients who recurred, 40 patients (75.5%) were HER3-positive. Thus, of the RTKs studied, HER3 was the only RTK identified as an independent prognostic indicator of stage II/III advanced gastric cancer with standard treatment.

  8. Immunohistochemical analysis of human brain suggests pathological synergism of Alzheimer's disease and diabetes mellitus.

    PubMed

    Valente, Tony; Gella, Alejandro; Fernàndez-Busquets, Xavier; Unzeta, Mercedes; Durany, Nuria

    2010-01-01

    It has been extensively reported that diabetes mellitus (DM) patients have a higher risk of developing Alzheimer's disease (AD), but a mechanistic connection between both pathologies has not been provided so far. Carbohydrate-derived advanced glycation endproducts (AGEs) have been implicated in the chronic complications of DM and have been reported to play an important role in the pathogenesis of AD. The earliest histopathological manifestation of AD is the apparition of extracellular aggregates of the amyloid beta peptide (Abeta). To investigate possible correlations between AGEs and Abeta aggregates with both pathologies, we have performed an immuhistochemical study in human post-mortem samples of AD, AD with diabetes (ADD), diabetic and nondemented controls. ADD brains showed increased number of Abeta dense plaques and receptor for AGEs (RAGE)-positive and Tau-positive cells, higher AGEs levels and major microglial activation, compared to AD brain. Our results indicate that ADD patients present a significant increase of cell damage through a RAGE-dependent mechanism, suggesting that AGEs may promote the generation of an oxidative stress vicious cycle, which can explain the severe progression of patients with both pathologies.

  9. Deciphering protein signatures using color, morphological, and topological analysis of immunohistochemically stained human tissues

    NASA Astrophysics Data System (ADS)

    Zerhouni, Erwan; Prisacari, Bogdan; Zhong, Qing; Wild, Peter; Gabrani, Maria

    2016-03-01

    Images of tissue specimens enable evidence-based study of disease susceptibility and stratification. Moreover, staining technologies empower the evidencing of molecular expression patterns by multicolor visualization, thus enabling personalized disease treatment and prevention. However, translating molecular expression imaging into direct health benefits has been slow. Two major factors contribute to that. On the one hand, disease susceptibility and progression is a complex, multifactorial molecular process. Diseases, such as cancer, exhibit cellular heterogeneity, impeding the differentiation between diverse grades or types of cell formations. On the other hand, the relative quantification of the stained tissue selected features is ambiguous, tedious and time consuming, prone to clerical error, leading to intra- and inter-observer variability and low throughput. Image analysis of digital histopathology images is a fast-developing and exciting area of disease research that aims to address the above limitations. We have developed a computational framework that extracts unique signatures using color, morphological and topological information and allows the combination thereof. The integration of the above information enables diagnosis of disease with AUC as high as 0.97. Multiple staining show significant improvement with respect to most proteins, and an AUC as high as 0.99.

  10. Biologic and immunohistochemical analysis of interleukin-6 expression in vivo. Constitutive and induced expression in murine polymorphonuclear and mononuclear phagocytes.

    PubMed Central

    Terebuh, P. D.; Otterness, I. G.; Strieter, R. M.; Lincoln, P. M.; Danforth, J. M.; Kunkel, S. L.; Chensue, S. W.

    1992-01-01

    Interleukin-6 (IL-6) is considered an important multifunctional cytokine involved in the regulation of a variety of cellular responses, including the induction of acute-phase protein synthesis, lymphocyte activation, and hematopoiesis. In vitro studies have identified many cells that can produce IL-6, but the cellular sources under physiologic conditions have yet to be identified. Using immunoaffinity purified goat anti-murine IL-6, the authors performed immunohistochemical studies to localize cells expressing IL-6 in selected organs of normal and endotoxin challenged NIH-Swiss outbred mice. In the blood, findings were correlated with cell-associated bioactivity using the standard B9 cell proliferation assay. In normal mice, constitutive expression was seen in granulocytes, monocytes and their precursors as well as in bone marrow and splenic stromal macrophages. Hepatic macrophages were negative, as were lymphocytes, megakaryocytes, erythroid precursors, and endothelial cells. In the absence of significant serum levels of IL-6, cell-associated IL-6 bioactivity was detected in circulating polymorphonuclear leukocytes (PMNs), but not lymphocytes. After endotoxin challenge, there was a threefold increase in PMN IL-6 content from 1 to 3 hours followed by almost complete depletion at 6 hours. This correlated well with a threefold increase of IL-6 mRNA in the bone marrow followed by a decrease at 6 hours. This pattern also correlated with serum levels of IL-6, which peaked at 3 hours and dropped significantly by 6 hours. By 24 hours, cell-associated IL-6 showed recovery with no increase in serum levels. In vivo findings showing IL-6 expression in bone marrow macrophages support in vitro studies suggesting a role for IL-6 in hematopoiesis. Furthermore, PMNs as well as macrophages are likely important sources of IL-6 during inflammatory and septic states. Images Figure 1 Figure 5 PMID:1372159

  11. Immunohistochemical Analysis of ATRX, IDH1 and p53 in Glioblastoma and Their Correlations with Patient Survival

    PubMed Central

    2016-01-01

    Glioblastoma (GBM) can be classified into molecular subgroups, on the basis of biomarker expression. Here, we classified our cohort of 163 adult GBMs into molecular subgroups according to the expression of proteins encoded by genes of alpha thalassemia/mental retardation syndrome X-linked (ATRX), isocitrate dehydrogenase (IDH) and TP53. We focused on the survival rate of molecular subgroups, depending on each and various combination of these biomarkers. ATRX, IDH1 and p53 protein expression were evaluated immunohistochemically and Kaplan-Meier analysis were carried out in each group. A total of 15.3% of enrolled GBMs demonstrated loss of ATRX expression (ATRX-), 10.4% expressed an aberrant IDH1 R132H protein (IDH1+), and 48.4% exhibited p53 overexpression (p53+). Survival differences were statistically significant when single protein expression or different combinations of expression of these proteins were analyzed. In conclusion, in the case of single protein expression, the patients with each IDH1+, or ATRX-, or p53- GBMs showed better survival than patients with counterparts protein expressed GBMs. In the case of double protein pairs, the patients with ATRX-/p53-, ATRX-/IDH1+, and IDH1+/p53- GBMs revealed better survival than the patients with GBMs with the remained pairs. In the case of triple protein combinations, the patients with ATRX-/p53-/IDH+ showed statistically significant survival gain than the patients with remained combination of proteins-expression status. Therefore, these three biomarkers, individually and as a combination, can stratify GBMs into prognostically relevant subgroups and have strong prognostic values in adult GBMs. PMID:27478330

  12. Immunohistochemical Analysis of ATRX, IDH1 and p53 in Glioblastoma and Their Correlations with Patient Survival.

    PubMed

    Chaurasia, Ajay; Park, Sung-Hye; Seo, Jeong-Wook; Park, Chul-Kee

    2016-08-01

    Glioblastoma (GBM) can be classified into molecular subgroups, on the basis of biomarker expression. Here, we classified our cohort of 163 adult GBMs into molecular subgroups according to the expression of proteins encoded by genes of alpha thalassemia/mental retardation syndrome X-linked (ATRX), isocitrate dehydrogenase (IDH) and TP53. We focused on the survival rate of molecular subgroups, depending on each and various combination of these biomarkers. ATRX, IDH1 and p53 protein expression were evaluated immunohistochemically and Kaplan-Meier analysis were carried out in each group. A total of 15.3% of enrolled GBMs demonstrated loss of ATRX expression (ATRX-), 10.4% expressed an aberrant IDH1 R132H protein (IDH1+), and 48.4% exhibited p53 overexpression (p53+). Survival differences were statistically significant when single protein expression or different combinations of expression of these proteins were analyzed. In conclusion, in the case of single protein expression, the patients with each IDH1+, or ATRX-, or p53- GBMs showed better survival than patients with counterparts protein expressed GBMs. In the case of double protein pairs, the patients with ATRX-/p53-, ATRX-/IDH1+, and IDH1+/p53- GBMs revealed better survival than the patients with GBMs with the remained pairs. In the case of triple protein combinations, the patients with ATRX-/p53-/IDH+ showed statistically significant survival gain than the patients with remained combination of proteins-expression status. Therefore, these three biomarkers, individually and as a combination, can stratify GBMs into prognostically relevant subgroups and have strong prognostic values in adult GBMs. PMID:27478330

  13. Immunohistochemical Analysis of Brainstem Lesions in the Autopsy Cases with Severe Motor and Intellectual Disabilities Showing Sudden Unexplained Death

    PubMed Central

    Hayashi, Masaharu; Sakuma, Hiroshi

    2016-01-01

    It is known that patients with severe motor and intellectual disabilities (SMID) showed sudden unexplained death (SUD), in which autopsy failed to identify causes of death. Although the involvement of brainstem dysfunction is speculated, the detailed neuropathological analysis still remains to be performed. In order to clarify pathogenesis, we investigated the brainstem functions in autopsy cases of SMID showing SUD. We immunohistochemically examined expressions of tyrosine hydroxylase, tryptophan hydroxylase, substance P, methionine-enkephalin, and c-fos in the serial sections of the midbrain, pons, and medulla oblongata in eight SUD cases and seven controls, having neither unexplained death nor pathological changes in the brain. Expressions of tyrosine hydroxylase and tryptophan hydroxylase were reduced in two of eight cases, and those of substance P and/or methionine-enkephalin were augmented in the pons and medulla oblongata in seven of eight cases, including the aforementioned two cases, when compared with those in controls. The hypoglossal nucleus and/or the dorsal vagal nucleus demonstrated increased neuronal immunoreactivity for c-fos in seven of eight cases, although there was no neuronal loss or gliosis in both the nuclei. Controls rarely showed immunoreactivity for c-fos in the medulla oblongata. These data suggest the possible involvement of brainstem dysfunction in SUD in patients with SMID, and consecutive neurophysiological evaluation of brainstem functions, such as all-night polysomnography and blink reflex, may be useful for the prevention of SUD, because some parameters in the neurophysiological examination are known to be related to the brainstem catecholamine neurons and the spinal tract nucleus of trigeminal nerve. PMID:27445960

  14. Immunohistochemical Analysis of ATRX, IDH1 and p53 in Glioblastoma and Their Correlations with Patient Survival.

    PubMed

    Chaurasia, Ajay; Park, Sung-Hye; Seo, Jeong-Wook; Park, Chul-Kee

    2016-08-01

    Glioblastoma (GBM) can be classified into molecular subgroups, on the basis of biomarker expression. Here, we classified our cohort of 163 adult GBMs into molecular subgroups according to the expression of proteins encoded by genes of alpha thalassemia/mental retardation syndrome X-linked (ATRX), isocitrate dehydrogenase (IDH) and TP53. We focused on the survival rate of molecular subgroups, depending on each and various combination of these biomarkers. ATRX, IDH1 and p53 protein expression were evaluated immunohistochemically and Kaplan-Meier analysis were carried out in each group. A total of 15.3% of enrolled GBMs demonstrated loss of ATRX expression (ATRX-), 10.4% expressed an aberrant IDH1 R132H protein (IDH1+), and 48.4% exhibited p53 overexpression (p53+). Survival differences were statistically significant when single protein expression or different combinations of expression of these proteins were analyzed. In conclusion, in the case of single protein expression, the patients with each IDH1+, or ATRX-, or p53- GBMs showed better survival than patients with counterparts protein expressed GBMs. In the case of double protein pairs, the patients with ATRX-/p53-, ATRX-/IDH1+, and IDH1+/p53- GBMs revealed better survival than the patients with GBMs with the remained pairs. In the case of triple protein combinations, the patients with ATRX-/p53-/IDH+ showed statistically significant survival gain than the patients with remained combination of proteins-expression status. Therefore, these three biomarkers, individually and as a combination, can stratify GBMs into prognostically relevant subgroups and have strong prognostic values in adult GBMs.

  15. Microscopy image analysis of p63 immunohistochemically stained laryngeal cancer lesions for predicting patient 5-year survival.

    PubMed

    Ninos, Konstantinos; Kostopoulos, Spiros; Kalatzis, Ioannis; Sidiropoulos, Konstantinos; Ravazoula, Panagiota; Sakellaropoulos, George; Panayiotakis, George; Economou, George; Cavouras, Dionisis

    2016-01-01

    The aim of the present study was to design a microscopy image analysis (MIA) system for predicting the 5-year survival of patients with laryngeal squamous cell carcinoma, employing histopathology images of lesions, which had been immunohistochemically (IHC) stained for p63 expression. Biopsy materials from 42 patients, with verified laryngeal cancer and follow-up, were selected from the archives of the University Hospital of Patras, Greece. Twenty six patients had survived more than 5 years and 16 less than 5 years after the first diagnosis. Histopathology images were IHC stained for p63 expression. Images were first processed by a segmentation method for isolating the p63-expressed nuclei. Seventy-seven features were evaluated regarding texture, shape, and physical topology of nuclei, p63 staining, and patient-specific data. Those features, the probabilistic neural network classifier, the leave-one-out (LOO), and the bootstrap cross-validation methods, were used to design the MIA-system for assessing the 5-year survival of patients with laryngeal cancer. MIA-system accuracy was about 90 % and 85 %, employing the LOO and the Bootstrap methods, respectively. The image texture of p63-expressed nuclei appeared coarser and contained more edges in the 5-year non-survivor group. These differences were at a statistically significant level (p < 0.05). In conclusion, this study has proposed an MIA-system that may be of assistance to physicians, as a second opinion tool in assessing the 5-year survival of patients with laryngeal cancer, and it has revealed useful information regarding differences in nuclei texture between 5-year survivors and non-survivors. PMID:26285779

  16. Immunohistochemical Analysis of Brainstem Lesions in the Autopsy Cases with Severe Motor and Intellectual Disabilities Showing Sudden Unexplained Death.

    PubMed

    Hayashi, Masaharu; Sakuma, Hiroshi

    2016-01-01

    It is known that patients with severe motor and intellectual disabilities (SMID) showed sudden unexplained death (SUD), in which autopsy failed to identify causes of death. Although the involvement of brainstem dysfunction is speculated, the detailed neuropathological analysis still remains to be performed. In order to clarify pathogenesis, we investigated the brainstem functions in autopsy cases of SMID showing SUD. We immunohistochemically examined expressions of tyrosine hydroxylase, tryptophan hydroxylase, substance P, methionine-enkephalin, and c-fos in the serial sections of the midbrain, pons, and medulla oblongata in eight SUD cases and seven controls, having neither unexplained death nor pathological changes in the brain. Expressions of tyrosine hydroxylase and tryptophan hydroxylase were reduced in two of eight cases, and those of substance P and/or methionine-enkephalin were augmented in the pons and medulla oblongata in seven of eight cases, including the aforementioned two cases, when compared with those in controls. The hypoglossal nucleus and/or the dorsal vagal nucleus demonstrated increased neuronal immunoreactivity for c-fos in seven of eight cases, although there was no neuronal loss or gliosis in both the nuclei. Controls rarely showed immunoreactivity for c-fos in the medulla oblongata. These data suggest the possible involvement of brainstem dysfunction in SUD in patients with SMID, and consecutive neurophysiological evaluation of brainstem functions, such as all-night polysomnography and blink reflex, may be useful for the prevention of SUD, because some parameters in the neurophysiological examination are known to be related to the brainstem catecholamine neurons and the spinal tract nucleus of trigeminal nerve. PMID:27445960

  17. Immunohistochemical analysis of a ruptured basilar top aneurysm autopsied 22 years after embolization with Guglielmi detachable coils

    PubMed Central

    Yuki, Ichiro; Spitzer, Daniel; Guglielmi, Guido; Duckwiler, Gary; Fujimoto, Motoaki; Takao, Hiroyuki; Jahan, Reza; Tateshima, Satoshi; Murayama, Yuichi; Vinuela, Fernando

    2014-01-01

    The authors report on the histologic and immunohistochemical analyses of a cerebral aneurysm embolized with platinum coils and with the longest observation period. A 58-year-old woman presenting with subarachnoid hemorrhage due to ruptured basilar top aneurysm was treated with Guglielmi detachable coils (GDC) 22 years ago. She was the 15th case since the GDC was introduced. After she died of unrelated causes, an autopsy and thorough histologic examination were performed. Gross examination revealed no adhesion between the aneurysm wall and the surrounding brain tissue. Histologic and immunohistochemical analyses demonstrated that the cavity of the aneurysm was filled with homogeneous collagenous fibrous tissue, while the neck was completely covered by a dense collagenous neointima and a smooth muscle cell layer. The unique histologic results of this case may contribute to a better understanding of the long-term evolution of the healing process in intracranial aneurysms successfully treated with the GDC. PMID:25056301

  18. Immunohistochemical analysis of a ruptured basilar top aneurysm autopsied 22 years after embolization with Guglielmi detachable coils.

    PubMed

    Yuki, Ichiro; Spitzer, Daniel; Guglielmi, Guido; Duckwiler, Gary; Fujimoto, Motoaki; Takao, Hiroyuki; Jahan, Reza; Tateshima, Satoshi; Murayama, Yuichi; Vinuela, Fernando

    2015-08-01

    The authors report on the histologic and immunohistochemical analyses of a cerebral aneurysm embolized with platinum coils and with the longest observation period. A 58-year-old woman presenting with subarachnoid hemorrhage due to ruptured basilar top aneurysm was treated with Guglielmi detachable coils (GDC) 22 years ago. She was the 15th case since the GDC was introduced. After she died of unrelated causes, an autopsy and thorough histologic examination were performed. Gross examination revealed no adhesion between the aneurysm wall and the surrounding brain tissue. Histologic and immunohistochemical analyses demonstrated that the cavity of the aneurysm was filled with homogeneous collagenous fibrous tissue, while the neck was completely covered by a dense collagenous neointima and a smooth muscle cell layer. The unique histologic results of this case may contribute to a better understanding of the long-term evolution of the healing process in intracranial aneurysms successfully treated with the GDC.

  19. Expression profiles of inhibitor of growth protein 2 in normal and cancer tissues: An immunohistochemical screening analysis.

    PubMed

    Zhao, Shuang; Yang, Xue-Feng; Gou, Wen-Feng; Lu, Hang; Li, Hua; Zhu, Zhi-Tu; Sun, Hong-Zhi; Zheng, Hua-Chuan

    2016-02-01

    Inhibitor of growth protein 2 (ING2) has an important role in the regulation of chromatin remodeling, cell proliferation, cell‑cycle arrest, senescence and apoptosis. The present study performed an immunohistochemical analysis for expression profiling of ING2 protein in an array of tissues comprising normal mouse and human tissues, as well as human hepatocellular (n=62), renal clear cell (n=62), pancreatic (n=62), esophageal squamous cell (n=45), cervical squamous cell (n=31), breast (n=144), gastric (n=196), colorectal (n=96), ovarian (n=208), endometrial (n=96) and lung (n=192) carcinoma tissues. In mouse tissues, ING2 was detected in the nuclei and cytoplasm of the glandular epithelium of breast, hepatocytes, intestine, bronchium and alveoli, as well as the squamous epithelium of skin and glomeruli, and in myocardial cells, while it was located in the cytoplasm of renal tubules and striated muscle cells. ING2 protein was scattered in the brain and spleen. In human tissues, ING2 protein was principally distributed in the cytoplasm, while in it was present in the cytoplasm and nuclei in the stomach, intestine, cervix, endometrium trachea, breast and pancreas. The nuclear location of ING2 in the stomach was more prominent than that in the cytoplasm. High ING2 immunoreactivity was detected in the tongue, stomach, skin, pancreas, cervix and breast, whereas weakly in the brain stem, thymus, thyroid, lung, striated muscle, testis, bladder and ovary. In total, 617 out of 1,194 of the tested cancer tissues (51.7%) were ING2-positive. In most cases, ING2 expression was found to be restricted to the cytoplasm of all cancer tissues, while in certain cancer types, including renal clear cell, ovarian and colorectal carcinoma, it was occasionally present in the nuclei. Among the cancer tissues examined, ING2 was most frequently expressed in breast cancer (67.4%) and gynecological cancer types, including ovarian cancer (61.5%) and endometrial cancer (57.3%). Compared with

  20. Expression profiles of inhibitor of growth protein 2 in normal and cancer tissues: An immunohistochemical screening analysis.

    PubMed

    Zhao, Shuang; Yang, Xue-Feng; Gou, Wen-Feng; Lu, Hang; Li, Hua; Zhu, Zhi-Tu; Sun, Hong-Zhi; Zheng, Hua-Chuan

    2016-02-01

    Inhibitor of growth protein 2 (ING2) has an important role in the regulation of chromatin remodeling, cell proliferation, cell‑cycle arrest, senescence and apoptosis. The present study performed an immunohistochemical analysis for expression profiling of ING2 protein in an array of tissues comprising normal mouse and human tissues, as well as human hepatocellular (n=62), renal clear cell (n=62), pancreatic (n=62), esophageal squamous cell (n=45), cervical squamous cell (n=31), breast (n=144), gastric (n=196), colorectal (n=96), ovarian (n=208), endometrial (n=96) and lung (n=192) carcinoma tissues. In mouse tissues, ING2 was detected in the nuclei and cytoplasm of the glandular epithelium of breast, hepatocytes, intestine, bronchium and alveoli, as well as the squamous epithelium of skin and glomeruli, and in myocardial cells, while it was located in the cytoplasm of renal tubules and striated muscle cells. ING2 protein was scattered in the brain and spleen. In human tissues, ING2 protein was principally distributed in the cytoplasm, while in it was present in the cytoplasm and nuclei in the stomach, intestine, cervix, endometrium trachea, breast and pancreas. The nuclear location of ING2 in the stomach was more prominent than that in the cytoplasm. High ING2 immunoreactivity was detected in the tongue, stomach, skin, pancreas, cervix and breast, whereas weakly in the brain stem, thymus, thyroid, lung, striated muscle, testis, bladder and ovary. In total, 617 out of 1,194 of the tested cancer tissues (51.7%) were ING2-positive. In most cases, ING2 expression was found to be restricted to the cytoplasm of all cancer tissues, while in certain cancer types, including renal clear cell, ovarian and colorectal carcinoma, it was occasionally present in the nuclei. Among the cancer tissues examined, ING2 was most frequently expressed in breast cancer (67.4%) and gynecological cancer types, including ovarian cancer (61.5%) and endometrial cancer (57.3%). Compared with

  1. MAGE-A is More Highly Expressed Than NY-ESO-1 in a Systematic Immunohistochemical Analysis of 3668 Cases.

    PubMed

    Kerkar, Sid P; Wang, Zeng-Feng; Lasota, Jerzy; Park, Tristen; Patel, Krishna; Groh, Eric; Rosenberg, Steven A; Miettinen, Markku M

    2016-05-01

    Two cancer testis antigens, the New York esophageal squamous cell carcinoma-1 (NY-ESO-1) and the melanoma-antigen family A (MAGE-A), represent promising immunotherapy targets due to the low expression of these antigens in nonmalignant tissue. To assess overexpression patterns in various cancers, we performed a systematic immunohistochemical analysis for NY-ESO-1 and MAGE-A on tissue array samples of 3668 common epithelial carcinomas (CA) and germ cell tumors of high prevalence and mortality. Here, we find significantly higher expression of MAGE-A (>50% on tumor cells) compared with NY-ESO-1 in several CAs including cutaneous squamous cell carcinomas (SCC) (52.8%/2.8%), esophageal SCC (50%/0%), head and neck SCC (41.1%/<1%), bladder urothelial CA (40.4%/8.3%), cervical/anal SCC (37.5%/0%), lung SCC (34%/3.8%), lung adenocarcinomas (27.6%/3.9%), ovarian CA (26.4%/3.6%), endometrial CA (26.3%/1.3%), lung small cell CA (24.4%/2.4%), gastric adenocarcinomas (20%/4%), breast mucinous CA (19.3%/0%), hepatocellular CA (18.8%/1.2%), breast infiltrating ductal CA (16.4%/1.8%), colorectal adenocarcinomas (10.7%/<1%), cholangiocarcinomas (9.8%/0%), thymic CA (9%/4.5%), and mesotheliomas (7.9%/<1%). Furthermore, high expression of MAGE-A, but not NY-ESO-1, was seen in whole slide evaluations of an independent cohort of metastatic SCC (45.5%/3.6%) and metastatic CA (13.5%/0%) of various primaries with significantly higher expression of MAGE-A in metastatic SCC compared with other metastatic CA. MAGE-A is also more highly expressed in germ cell tumors, seminomas (69%/3.5%) and nonseminomas (40.1%/4.7%). In summary, MAGE-A is more highly expressed than NY-ESO-1 in a majority of human malignancies, and targeting MAGE-A may benefit a large number of patients.

  2. Immunohistochemical Analysis of PD-L1 Expression in Canine Malignant Cancers and PD-1 Expression on Lymphocytes in Canine Oral Melanoma.

    PubMed

    Maekawa, Naoya; Konnai, Satoru; Okagawa, Tomohiro; Nishimori, Asami; Ikebuchi, Ryoyo; Izumi, Yusuke; Takagi, Satoshi; Kagawa, Yumiko; Nakajima, Chie; Suzuki, Yasuhiko; Kato, Yukinari; Murata, Shiro; Ohashi, Kazuhiko

    2016-01-01

    Spontaneous cancers are common diseases in dogs. Among these, some malignant cancers such as oral melanoma, osteosarcoma, hemangiosarcoma, and mast cell tumor are often recognized as clinical problems because, despite their high frequencies, current treatments for these cancers may not always achieve satisfying outcomes. The absence of effective systemic therapies against these cancers leads researchers to investigate novel therapeutic modalities, including immunotherapy. Programmed death 1 (PD-1) is a costimulatory receptor with immunosuppressive function. When it binds its ligands, PD-ligand 1 (PD-L1) or PD-L2, PD-1 on T cells negatively regulates activating signals from the T cell receptor, resulting in the inhibition of the effector function of cytotoxic T lymphocytes. Aberrant PD-L1 expression has been reported in many human cancers and is considered an immune escape mechanism for cancers. In clinical trials, anti-PD-1 or anti-PD-L1 antibodies induced tumor regression for several malignancies, including advanced melanoma, non-small cell lung carcinoma, and renal cell carcinoma. In this study, to assess the potential of the PD-1/PD-L1 axis as a novel therapeutic target for canine cancer immunotherapy, immunohistochemical analysis of PD-L1 expression in various malignant cancers of dogs was performed. Here, we show that dog oral melanoma, osteosarcoma, hemangiosarcoma, mast cell tumor, mammary adenocarcinoma, and prostate adenocarcinoma expressed PD-L1, whereas some other types of cancer did not. In addition, PD-1 was highly expressed on tumor-infiltrating lymphocytes obtained from oral melanoma, showing that lymphocytes in this cancer type might have been functionally exhausted. These results strongly encourage the clinical application of PD-1/PD-L1 inhibitors as novel therapeutic agents against these cancers in dogs. PMID:27276060

  3. Immunohistochemical Analysis of PD-L1 Expression in Canine Malignant Cancers and PD-1 Expression on Lymphocytes in Canine Oral Melanoma

    PubMed Central

    Maekawa, Naoya; Konnai, Satoru; Okagawa, Tomohiro; Nishimori, Asami; Ikebuchi, Ryoyo; Izumi, Yusuke; Takagi, Satoshi; Kagawa, Yumiko; Nakajima, Chie; Suzuki, Yasuhiko; Kato, Yukinari; Murata, Shiro; Ohashi, Kazuhiko

    2016-01-01

    Spontaneous cancers are common diseases in dogs. Among these, some malignant cancers such as oral melanoma, osteosarcoma, hemangiosarcoma, and mast cell tumor are often recognized as clinical problems because, despite their high frequencies, current treatments for these cancers may not always achieve satisfying outcomes. The absence of effective systemic therapies against these cancers leads researchers to investigate novel therapeutic modalities, including immunotherapy. Programmed death 1 (PD-1) is a costimulatory receptor with immunosuppressive function. When it binds its ligands, PD-ligand 1 (PD-L1) or PD-L2, PD-1 on T cells negatively regulates activating signals from the T cell receptor, resulting in the inhibition of the effector function of cytotoxic T lymphocytes. Aberrant PD-L1 expression has been reported in many human cancers and is considered an immune escape mechanism for cancers. In clinical trials, anti-PD-1 or anti-PD-L1 antibodies induced tumor regression for several malignancies, including advanced melanoma, non-small cell lung carcinoma, and renal cell carcinoma. In this study, to assess the potential of the PD-1/PD-L1 axis as a novel therapeutic target for canine cancer immunotherapy, immunohistochemical analysis of PD-L1 expression in various malignant cancers of dogs was performed. Here, we show that dog oral melanoma, osteosarcoma, hemangiosarcoma, mast cell tumor, mammary adenocarcinoma, and prostate adenocarcinoma expressed PD-L1, whereas some other types of cancer did not. In addition, PD-1 was highly expressed on tumor-infiltrating lymphocytes obtained from oral melanoma, showing that lymphocytes in this cancer type might have been functionally exhausted. These results strongly encourage the clinical application of PD-1/PD-L1 inhibitors as novel therapeutic agents against these cancers in dogs. PMID:27276060

  4. Extragonadal malignant germ cell tumors: a clinicopathological and immunohistochemical analysis of 48 cases at a single Chinese institution.

    PubMed

    Gao, Yuping; Jiang, Jiyao; Liu, Qiang

    2015-01-01

    Primary extragonadal malignant germ cell tumors (EMGCTs) are rare and characterized by the location in the midline of the body, including mediastinum, CNS, retroperitoneum and coccyx. EMGCTs present with different clinical and biologic characteristics in different tumor locations. Accurately diagnosing MEGCTs would be very difficult by performing on HE staining alone, and requires immunohistochemical verification. This study was to investigate the biological feature of EMGCTs and diagnostic value of immunohistochemical markers OCT3/4, CD117, PLAP, AFP, β-HCG and CD30 in EMGCTs. A retrospective study was performed on 48 patients with EMGCTs. EMGCTs were found to occur predominantly in males, especially for mediastinal MGCTs. The tumor locations included mediastinum, CNS and retroperitoneum. The mediastinum and CNS were the most common sites of EMGCTs. Seminoma/germinomas (64.6%) was the most common histological subtypes of EMGCTs. Chest pain, dyspnea, cough and fever were the most common clinical presentations in mediastinal MGCTs. Headache, visual disturbances, endocrine abnormalities, and signs of increased intracranial pressure were common clinical symptoms in CNS MGCTs. Abdominal mass with or without pain, backache and weight loss were common clinical presentations in retroperitoneal MGCTs. PLAP, CD117 and OCT3/4 were highly expressed in seminomas/gernimomas. CD30, EMA and CK AE1/3 staining were positive in embryonal carcinoma. AFP and β-HCG positive staining are characteristic in yolk sac tumors and choriocarcinoma, respectively. Patients with seminomas/germinomas had a better prognosis than those with NS/G-GCTs. Our finding suggests that the accurate diagnosis of EMGCTs is critical not only for predicting the tumor progression but also for patient management. Immunohistochemical markers have become an important tool in the diagnosis and differential diagnosis of EMGCTs.

  5. Immunohistochemical characterization of brain-invasive meningiomas

    PubMed Central

    Backer-Grøndahl, Thomas; Moen, Bjørnar H; Arnli, Magnus B; Torseth, Kathrin; Torp, Sverre H

    2014-01-01

    Brain-invasive meningiomas have an adverse prognosis, so it is important to detect and correctly evaluate brain invasion by light microscopy. Furthermore, the underlying biological mechanisms responsible for brain-invasive growth are incompletely understood. The primary aim of this study was to identify immunohistochemical markers that could improve identification and evaluation of brain invasion in meningiomas. A second aim was to investigate the process of brain invasion using immunohistochemical markers of proliferation, extracellular matrix modulation, and cell adhesion. From a series of 196 human meningiomas, 67 cases were selected for analysis because of the presence of brain tissue in tumor specimens. Fourteen of these 67 meningiomas were brain-invasive. Invasiveness was determined primarily by evaluation of hematoxylin-erytrosin-saffron- (HES-) stained specimens, although glial fibrillary acidic protein (GFAP), anti-collagen IV, and cluster of differentiation 44 (CD44) markers provided additional information. It was important to examine microscopic sections from various levels of the paraffin-embedded tissue block to adequately assess invasiveness. Sections stained using antibodies against Ki-67/MIB-1, phospohistone-H3 (PHH3), matrix metalloproteinase-9 (MMP-9), cathepsin D, plasminogen activator inhibitor-1 (PAI-1), and E-cadherin antigens were used to characterize brain-invasive meningiomas and to investigate the process of brain invasion. Only increased expression of the extracellular matrix modulator MMP-9 correlated with brain-invasive growth (p=0.025). Examination of HES-stained sections identified brain invasion. Use of relevant immunohistochemical markers did not contribute substantially to this evaluation. Evaluation of stepwise sections should be considered when brain-invasive growth is suspected. MMP-9 may be an important mediator of brain-invasive growth. PMID:25400818

  6. Lipomatous hemangiopericytoma: a fat-containing variant of solitary fibrous tumor? Clinicopathologic, immunohistochemical, and ultrastructural analysis of a series in favor of a unifying concept.

    PubMed

    Guillou, L; Gebhard, S; Coindre, J M

    2000-09-01

    The dinicopathologic, immunohistochemical and ultrastructural features of 13 lipomatous hemangiopericytomas are presented. There were 6 male and 7 female patients whose ages at diagnosis ranged from 27 to 75 years (median 48) all presenting with a mass of variable duration. The tumor sizes ranged from 1.7 cm to 19 cm (median 5.5 cm). The locations included the orbit (1), neck (1), mediastinum (1), epicardium (1), retroperitoneum (3), right iliac fossa (1), and upper (1) and lower (4) extremity. Histologically, the lesions were composed of a varying admixture of spindle-shaped to round cells, variably collagenous stroma, adipose tissue, and branched, often thick-walled, hemangiopericytoma-like vessels. For 11 tumors, the mitotic activity ranged from 1 to 3 mitoses per 10 high-power fields (HPF). One tumor which contained hypercellular areas showed 13 mitoses per 10 HPF, and another hypercellular lesion showed up to 43 mitoses per 10 HPF, abnormal mitoses, and necrosis. Immunohistochemically, tumor cells were invariably positive for vimentin and CD99, and mostly for CD34 but negative for desmin, keratin, CD31, CD117 (c-kit), and inhibin. About half of the tumors showed reactivity for bcl-2. Occasionally, focal reactivity was also observed for smooth muscle actin, muscle-specific actin, S100 protein, and epithelial membrane antigen. Ultrastructural examination of seven cases showed features in keeping with fibroblastic, myofibroblastic, or pericytic differentiation. Treatment consisted of simple tumorectomy in 10 cases and wide excision in 3. Follow-up information on 10 patients (range: 6 to 77 months; median: 18 months) showed no recurrence. Lipomatous hemangiopericytoma which share the clinical, pathologic, immunohistochemical, and ultrastructural features of solitary fibrous tumor (SFT) is likely to represent, in most cases, a fat-containing variant of SFT.

  7. Immunohistochemical Expression of PCNA and CD34 in Colorectal Adenomas and Carcinomas Using Specified Automated Cellular Image Analysis System: A Clinicopathologic Study

    PubMed Central

    Qasim, Ban J.; Ali, Hussam H.; Hussein, Alaa G.

    2012-01-01

    Background/Aim: To evaluate the immunohistochemical expression of proliferating cell nuclear antigen (PCNA) and CD34 in colorectal adenomas and carcinomas, and to correlate this expression with different clinicopathologic parameters. Materials and Methods: The study was retrospectively designed. A total of 86 tissue samples, including 33 paraffin blocks from patients with colorectal adenomas, 33 paraffin blocks from patients with colorectal adenocarcinomas, and a control group of 20 samples of nontumerous colonic tissue, were included in the study. From each block, 3 sections of 5 ΅m thickness were taken, 1 section was stained with hematoxylin and eosin (H and E) and the other 2 sections were stained immunohistochemically for PCNA and CD34. Scoring of the immunohistochemical staining was performed using a specified automated cellular image analysis system (Digimizer). Results: PCNA expression was significantly increased in a sequence of normal mucosa–adenoma–carcinoma. It was significantly higher in adenomas ≥ 1 cm and those with severe dysplasia, and it showed a significant positive correlation with grade and lymph node involvement in colorectal carcinoma. CD34 showed significantly higher expression in carcinoma than adenoma and in adenoma than in the control group. CD34 expression showed a significant correlation with adenomas carrying severe dysplasia and large-sized adenomas (≥1cm). It was significantly correlated with tumor grade, lymphovascular invasion, and lymph node involvement in colorectal carcinoma. Conclusion: PCNA plays an important role in colorectal neoplastic progression and can be utilized as ancillary marker for the risk of malignant transformation in colorectal adenomas as it correlates with high grade dysplasia and size. Intratumoral quantification of the mean (A and N) of CD34 in colorectal carcinoma reflects the grade of tumors and can predict lymph node involvement and lymphovascular invasion, to make a useful additional prognostic

  8. [Immunohistochemical Analysis of Krebs von den Lungen-6 (KL-6) Expression in Lung Tissue in Primary Lung Cancer Patients with High Serum KL-6 Levels].

    PubMed

    Yatsuyanagi, Eiji; Sato, Kazuhiro; Sato, Keisuke

    2015-09-01

    We investigated sialylated carbohydrate antigen( Krebs von den Lungen-6:KL-6) expression in lung tissue and correlation between the expression and serum KL-6 level in the patients with primary lung cancer. Thirty-four primary lung cancer patients with high serum KL-6 levels( >500 U/ml) were evaluated. A coexistence of interstitial pneumonia (IP) was histopathologically evaluated and an immunohistochemical staining using a mouse anti-human KL-6 antibody (mKL-6) was performed. A multiple regression analysis was also caluculated using a serum KL-6 level as a target variable and the histopathological and immunohistochemical factors (KL-6 expression in cancer tissue and IP tissue, coexistence of IP, tumor size, pathological staging) as descriptive variables. Twenty-two patients (64.7%) were histopathologically concomitant with IP. Cancer tissues were positively stained by mKL-6 in 32 patients (94.1%). Among them, 20 patients were concomitant with IP and all of their cancer tissues were more strongly stained by mKL-6 than IP tissues. Although considerable high rate of lung cancer patients might express the KL-6 in the cancer tissue, we could not reveal the relationship between the expression and serum KL-6 level by a multiple regression analysis. For revealing the mechanism of elevating serum KL-6 level in the patients with lung cancer, more detailed and powerful study is thought to be needed.

  9. Immunohistochemical analysis of oxidative DNA damage in arsenic-related human skin samples from arsenic-contaminated area of China.

    PubMed

    An, Yan; Gao, Zenglin; Wang, Zhongwen; Yang, Shaohe; Liang, Jianfen; Feng, Yan; Kato, Koichi; Nakano, Masayuki; Okada, Shoji; Yamanaka, Kenzo

    2004-10-01

    The appearance of 8-oxo-2'-deoxyguanosine (8-oxodG) was examined immunohistochemically using an 8-oxodG-monoclonal antibody in 28 cases of arsenic-related human skin tumors and in 20 cases of arsenic-unrelated human skin cancer to determine if the induction of oxidative stress participates in skin tumorigenesis caused by arsenics. The rate of 8-oxodG-positive was significantly higher in arsenic-related human skin cancer (28 of 28, 100%) than in arsenic-unrelated human skin cancer (3 of 20, 15%, P<0.01 by Chi2 test). Moreover, in all the arsenic-related skin samples, 8-oxodG was detected not only in tumor tissues but also in keratosis and normal tissues. These results suggest that the induction of oxidative stress may play an important role in arsenic carcinogenesis.

  10. A peripheral ameloblastic fibro-odontoma in a 3-year-old girl: case report, immunohistochemical analysis, and literature review.

    PubMed

    Lin, Yi-Chun; Hsu, Hsiu-Ming; Liu, Chiang-Shin; Yuan, Kuo

    2014-01-01

    Ameloblastic fibro-odontoma (AFO) predominantly occurs in the jaw bones of children and young adults. Extraosseous AFO is extremely rare. We describe a peripheral ameloblastic fibro-odontoma in the maxillary gingiva of a 3-year-old girl. The clinical appearance resembled fiery red reactive gingival lesions. The histopathological examination of the excised lesion showed small islands and cords of odontogenic epithelium with cellular myxoid stroma in the subepithelial tissue. The mass contained calcified material and an enamel-like deposit. Many small blood vessels appeared in the connective tissue surrounding the odontogenic epithelium. The immunohistochemical assays showed strong reactivity for amelogenin, β-catenin, CD44, and CD31 in the tissue sections. There was no recurrence after the 1-year follow-up. Because this lesion clinically resembles other nonneoplastic lesions and is very rare in gingiva, establishing a correct diagnosis is achieved only based on specific histological characteristics. Conservative excision of the tumor is the treatment of choice.

  11. Unusual mesenchymal and mixed tumors of the salivary gland. An immunohistochemical and flow cytometric analysis of three cases.

    PubMed

    Bocklage, T; Feddersen, R

    1995-01-01

    Histological, immunohistochemical, and flow cytometric characteristics of three unusual parotid gland tumors are described. The patients were adult white men with carcinoma ex pleomorphic adenoma, true malignant mixed tumor, and primary parotid gland chondrosarcoma. The carcinoma ex pleomorphic adenoma showed evidence of simultaneous epithelial, myoepithelial, and mesenchymal differentiation by immunohistochemistry. The true malignant mixed tumor exhibited variable positivity for two keratins, vimentin, proliferating cell nuclear antigen, Ki67, and p53. The chondrosarcoma initially stained for vimentin, S100, muscle-specific actin, proliferating cell nuclear antigen, and Ki67, but it lost actin expression in its first recurrence, accompanied by more extensive Ki67 staining. DNA ploidy varied from diploid to aneuploid with intratumoral variation in the carcinosarcoma. S-phase fractions ranged from 2.43% to 13.9%. The findings underscore the diversity of tumors that may be pathogenetically related to, and at times derived from, pleomorphic adenoma. PMID:7802557

  12. Immunohistochemical analysis of the distribution of measles related antigen in the intestinal mucosa in inflammatory bowel disease

    PubMed Central

    Iizuka, M; Chiba, M; Yukawa, M; Nakagomi, T; Fukushima, T; Watanabe, S; Nakagomi, O

    2000-01-01

    BACKGROUND—Measles virus is implicated in the aetiology of Crohn's disease. This measles hypothesis is mainly supported by immunohistochemical findings that the measles related antigen is present in the intestine of patients with Crohn's disease. Recently we isolated this antigen from the intestine of a patient with Crohn's disease using a molecular cloning technique and produced the monoclonal antibody against it (designated 4F12).
AIM—To discover whether the measles related antigen is uniquely present in Crohn's disease.
SUBJECTS/METHODS—Colonic mucosa samples from 20 patients with Crohn's disease, 20 with ulcerative colitis, 11 with non-inflammatory bowel disease (IBD) colitis, and nine controls were immunohistochemically stained with the anti-measles monoclonal antibody 4F12. The numbers of positive cells, the ratio of positive cells to nucleated cells, and the staining intensity of the positive cells were compared. Furthermore, the distribution of the measles antigen in other human organs was examined.
RESULTS—Both the number of positive cells and the ratio of positive cells to nucleated cells were significantly increased in Crohn's disease, ulcerative colitis, and non-IBD colitis compared with controls (p<0.05) but were similar among the three disease groups. The staining intensity of the positive cells was also similar among the three disease groups. Small numbers of positive cells were observed in the oesophagus, stomach, duodenum, jejunum, and lung.
CONCLUSIONS—The presence of the measles related antigen in the colonic mucosa was not unique to Crohn's disease. These results, together with the observation that such a measles related antigen was derived from host protein, do not support the hypothesis that measles virus causes Crohn's disease.


Keywords: Crohn's disease; measles virus; immunohistochemistry; ulcerative colitis; inflammatory bowel disease; molecular mimicry PMID:10644308

  13. Phenotype and cell proliferation activity of duct-like structures in human sublingual glands: a histological and immunohistochemical study

    PubMed Central

    TOLENTINO, Elen de Souza; TEIXEIRA, Cleverson Soares; AZEVEDO-ALANIS, Luciana Reis; HONÓRIO, Heitor Marques; DAMANTE, José Humberto

    2015-01-01

    There are several age-related microscopic changes in the salivary glands, including the increase in the number of duct-like structures (DLS). However, the true origin and the phenotype of the DLS are not known. Objective To evaluate the phenotype and the cell proliferation index of the DLS of human sublingual glands. Material and Methods Sixty sublingual glands obtained from human cadavers were divided into two groups - 0-30 and 61-90 years old. The phenotype was estimated by immunostaining for cytokeratin 19 (CK 19) and the S-100 protein as well as by the presence of mucin and glycogen. The cell proliferation index was determined by the Ki-67 antibody. The histochemical techniques used periodic acid-Schiff (PAS) and Alcian Blue. In each captured microscopic field, the DLS were counted to establish a percentage for the staining profile. The statistical analysis was accomplished using Student’s t-test, the Mann-Whitney test and Pearson’s correlation coefficient (p<0.05). Results Comparing both groups, only CK 19 showed a statistically significant difference (p=0.033), with the strongest expression in the elderly group. There was no significant difference between PAS and Alcian Blue (p=0.270). In both groups, the immunostaining for CK 19 was stronger than that for S-100 (p=0.004;p<0.001), but there was no correlation between the two immunomarkers (ρ=-0.163; p=0.315). There was no immunostaining for Ki-67. Conclusions DLS demonstrate a ductal phenotypic profile and do not present cell proliferation activity. DLS may represent a regressive process arising from acini or represent the result of metaplasia. PMID:26221919

  14. Immunohistochemical observations on tumor suppressor gene p53 status in mouse fibrosarcoma following in-vivo photodynamic therapy: the role of xanthine oxidase activity

    NASA Astrophysics Data System (ADS)

    Ziolkowski, Piotr P.; Symonowicz, Krzysztof; Milnerowicz, Artur; Osiecka, Beata J.

    1997-12-01

    Tumor suppressor gene p53 expression in a mouse fibrosarcoma following in-vivo photodynamic therapy has been studied using the immunohistochemical method. Photodynamic treatment involved injections of the well known sensitizer -- hematoporphyrin derivative at the doses 1.25 and 2.5 mg/kg of body weight and irradiations at the doses 25 and 50 J/sq cm. Glass slide preparations from PDT-treated tumors were obtained at different time points (15, 60 minutes, 2 and 24 hours) after therapy, subsequently stained for wild type/mutant p53, and assessed for positive reaction. High PDT doses (HpD -- 2.5 mg/kg; light dose -- 50 J/sq cm) correlated with decreased expression of p53 in tumor cells. The other part of the study was directed to measure the xanthine oxidase (XO) activity in the tumor cells. PDT included injections of HpD and light exposure at the same doses as for p53 study. We observed a complete inhibition of the enzyme activity. The slight increase in XO activity was found following treatment with either light or HpD alone.

  15. Immunohistochemical Analysis of Collagen IV and Laminin Expression in Spontaneous Melanoma Regression in the Melanoma-Bearing Libechov Minipig.

    PubMed

    Planska, Daniela; Burocziova, Monika; Strnadel, Jan; Horak, Vratislav

    2015-01-01

    Spontaneous regression (SR) of human melanoma is a rare, well-documented phenomenon that is not still fully understood. Its detailed study cannot be performed in patients due to ethical reasons. Using the Melanoma-bearing Libechov Minipig (MeLiM) animals of various ages (from 3 weeks to 8 months) we implemented a long-term monitoring of melanoma growth and SR. We focused on immunohistochemical detection of two important extracellular matrix proteins, collagen IV and laminin, which are associated with cancer. We showed that SR of melanoma is a highly dynamic process. The expression of collagen IV and laminin correlated with changes in population of melanoma cells. Tumours of 3-week-old animals consisted primarily of melanoma cells with a granular expression of collagen IV and laminin around them. Thereafter, melanoma cells were gradually destroyed and tumour tissue was rebuilt into the connective tissue. Collagen IV expression slightly increased in tumours of 10-week-old pigs showing extracellular fibrous appearance. In tumours of older animals, areas lacking melanoma cells demonstrated a low expression and areas still containing melanoma cells a high expression of both proteins. We considered the age of 10 weeks as a turning point in the transition between tumour growth and SR of the MeLiM melanoma.

  16. Really old-palaeoimmunology: immunohistochemical analysis of extracellular matrix proteins in historic and pre-historic material.

    PubMed

    Wick, G; Kalischnig, G; Maurer, H; Mayerl, C; Müller, P U

    2001-09-01

    In this review, we summarize data concerning the respective preservation and deterioration of antigenic determinants of various collagenous and non-collagenous extracellular matrix (ECM) proteins in palaeontologic material of different ages. ECM proteins are the major quantitative constituents of mammalian organisms and were, therefore, selected as important representative proteins for these analyses. The specimens, studied by immunofluorescence and immunohistochemical techniques, included the skin of 500-1500 year-old human mummies from Peru, skin and striated muscle from the 5300-year-old glacier mummy ("Iceman") from Tyrol, Austria, and a 50-million-year-old bat with preserved soft body parts from the fossil excavation site of Messel, Germany. In frozen sections of the former two sources, epitopes recognized by specific antibodies for triple-helical antigenic determinants of different types of collagen resistant against conventional proteases were preserved, while non-helical domains, as well as the non-collagenous ECM proteins, could no longer be demonstrated. The fossil bat, although showing evidence of fibrous, collagen-like structures in conventional histology, revealed no collagenous or non-collagenous ECM proteins by any technique. It later turned out that this was due to the replacement of the original soft parts in these fossils by lawns of bacteria. These studies introduced immunological techniques into palaeontology and opened new approaches for studying physiologically- and pathologically-altered structures in tissues of animals and humans of considerable historical age.

  17. Sialadenoma papilliferum: immunohistochemical study.

    PubMed

    Gomes, A P N; Sobral, A P V; Loducca, S V L; de Araújo, V C

    2004-09-01

    Sialadenoma papilliferum (SP) is a rare benign tumour of salivary gland origin, which has been included among the ductal papillomas in the latest classification of tumours by the World Health Organisation. Two SP from the minor salivary gland of the palate of middle age patients were presented and studied by immunohistochemical. Our results showed presence of cytokeratins (CKs) 13, 14, 7, 8, 19 and absence of vimentin and smooth muscle actin. This immunoprofile is similar to the excretory duct of salivary gland.

  18. Immunohistochemical analysis of the neural structures of the posterior cruciate ligament in osteoarthritis patients submitted to total knee arthroplasty: an analysis of thirty-four cases

    PubMed Central

    Martins, Glaucus Cajaty; Camanho, Gilberto; Rodrigues, Mara Ibis

    2015-01-01

    OBJECTIVES: Many authors recommend posterior cruciate ligament-retaining arthroplasty with the intention to maintain the proprioception properties of this ligament. Preservation of the neuroreceptors and nervous fibers may be essential for retaining the proprioception function of the posterior cruciate ligament. The present study was thus developed to evaluate the presence of neural structures in the posterior cruciate ligament resected during posterior stabilized arthroplasty in osteoarthritis patients. In particular, clinical, radiographic and histological parameters were correlated with the presence or absence of neural structures in the posterior cruciate ligament. METHODS: In total, 34 posterior cruciate ligament specimens were stained with hematoxylin-eosin and Gomori trichrome. An immunohistochemical analysis using antibodies against the S100 protein and neurofilaments was also performed. The presence of neural structures was correlated with parameters such as tibiofemoral angulation, histological degeneration of the posterior cruciate ligament, Ahlbäck radiological classification, age, gender and the histologic pattern of the synovial neurovascular bundle around the posterior cruciate ligament. RESULTS: In total, 67.5% of the cases presented neural structures in the posterior cruciate ligament. In 65% of the cases, the neurovascular bundle was degenerated. Nervous structures were more commonly detected in varus knees than in valgus knees (77% versus 50%). Additionally, severe histologic degeneration of the posterior cruciate ligament was related to neurovascular bundle degeneration. CONCLUSIONS: Severe posterior cruciate ligament degeneration was related to neurovascular bundle compromise. Neural structures were more commonly detected in varus knees. Intrinsic neural structures were detected in the majority of the posterior cruciate ligaments of patients submitted to knee arthroplasty for osteoarthritis. PMID:25789514

  19. The Y-organ secretory activity fluctuates in relation to seasons of molt and reproduction in the brachyuran crab, Metopograpsus messor (Grapsidae): Ultrastructural and immunohistochemical study.

    PubMed

    Shyamal, Sharmishtha; Sudha, K; Gayathri, N; Anilkumar, G

    2014-01-15

    This paper presents a first-time report on the localization, structure and seasonal secretory activity of the Y-organ of a grapsid brachyuran crab (Metopograpsus messor). Having exhibited discrete seasonality with reference to the programming of molt and reproduction, this brachyuran crab has offered us an excellent model to obtain a clear picture of the fluctuating secretory nature of the Yorgan, all the way through the reproductive (August-December) as well as the molt-reproduction active (January-May) and inactive (June-July) seasons. Ultrastructural studies revealed that the secretion of the Y-organ was at its peak in premolt crabs during molt-reproduction season (January-May). Interestingly, the Y-organs of the intermolt females that engaged in breeding activity showed higher levels of secretion than those of the molt-reproduction inactive season (June-July), implicating the gland's involvement in reproduction. Immunohistochemical studies using the antiserum raised against 2-succinyl conjugate of ecdysone have demonstrated the ecdysteroid nature of the secretion from the Y-organ, and results of the quantitative assay of ecdysteroids (through radioimmunoassay) revealed that the hormone titer fluctuates in consonance with the Y-organ's secretory activity during seasons of molt and reproduction. Pertinently, not only that the paper gives us a comprehensive understanding on the secretory activity of the Y-organ in a season-dependent fashion, it also allows us to have a better insight into the gland's function related to molting and reproduction (for the first time) in a grapsid brachyuran crab.

  20. Transferrin receptor expression in rat liver: immunohistochemical and biochemical analysis of the effect of age and iron storage.

    PubMed

    Sciot, R; Verhoeven, G; Van Eyken, P; Cailleau, J; Desmet, V J

    1990-03-01

    Hepatic transferrin receptors were studied in normal male rats at 1 to 59 wk after weaning, using immunohistochemical and biochemical techniques. The number of transferrin receptors measured and the intensity of the staining in situ decreased rapidly during the first 10 wk of life and more slowly thereafter. Immunohistochemistry further demonstrated changes in the topographical and (sub)cellular localization of the transferrin receptor. In the young rat livers, staining was almost exclusively present on hepatocytes in acinar zone 2 + 3 in a honeycomb to sinusoidal pattern. With aging, a panacinar heterogeneous and mainly sinusoidal staining of hepatocytes was more frequent. Kupffer cell positivity was more obvious as compared with the young rat livers. The observed changes in transferrin receptor expression may partly be explained by age-dependent alterations in DNA synthesis and proliferative potential of the liver cells. A series of rats were iron loaded with carbonyl iron up to 39 wk and "unloaded" by administration of a normal diet during 20 wk. In these animals, serial histochemical studies showed predominantly parenchymal (7 to 14 wk), mixed parenchymal and reticuloendothelial (39 wk) and almost exclusive reticuloendothelial siderosis (59 wk). In the siderotic livers transferrin receptor numbers tended to be lower than in the controls with significant differences after 14 and 39 wk. Immunohistochemistry showed decreased parenchymal but increased reticuloendothelial transferrin receptor expression with iron load. After the period of unloading, parenchymal transferrin receptors were virtually absent despite the negligible siderosis of these cells. In contrast, siderotic reticuloendothelial cells were intensely positive. These findings support down-regulation of parenchymal transferrin receptor resulting from iron storage. However, the positivity of siderotic reticuloendothelial cells and the absence of re-emergence of parenchymal receptors in conditions of

  1. Immunohistochemical analysis of MUC5B apomucin expression in breast cancer and non-malignant breast tissues.

    PubMed

    Sóñora, Cecilia; Mazal, Daniel; Berois, Nora; Buisine, Marie-Pierre; Ubillos, Luis; Varangot, Mario; Barrios, Enrique; Carzoglio, Julio; Aubert, Jean-Pierre; Osinaga, Eduardo

    2006-03-01

    A deregulation of several MUC genes (MUC1, MUC2, MUC3, MUC5AC, and MUC6) was previously demonstrated in breast carcinomas. Considering that recently we found the "non-mammary" MUC5B mRNA in primary breast tumors (Berois et al. 2003), we undertook the present study to evaluate the expression profile of MUC5B protein product in breast tissues, using LUM5B-2 antisera raised against sequences within the non-glycosylated regions of this apomucin. Expression of MUC5B by breast cancer cells was confirmed by immunocytochemistry, in situ hybridization, and Western blot on MCF-7 cancer cells. Using an immunohistochemical procedure, MUC5B apomucin was detected in 34/42 (81%) primary breast tumors, in 13/14 (92.8%) samples of non-malignant breast diseases, in 8/19 (42.1%) samples of normal-appearing breast epithelia adjacent to cancer, and in 0/5 normal control breast samples. The staining pattern of MUC5B was very different when comparing breast cancer cells (cytoplasmic) and non-malignant breast cells (predominantly apical and in the secretory material). We analyzed MUC5B mRNA expression using RT-PCR in bone marrow aspirates from 22/42 patients with breast cancer to compare with MUC5B protein expression in the primary tumors. Good correlation was observed because the six MUC5B-positive bone marrow samples also displayed MUC5B expression in the tumor. Our results show, for the first time at the protein level, that MUC5B apomucin is upregulated in breast cancer. Its characterization could provide new insights about the glycobiology of breast cancer cells. PMID:16148312

  2. Immunohistochemical and biochemical analysis of the oestrogen regulated protein pS2, and its relation with oestrogen receptor and progesterone receptor in breast cancer.

    PubMed Central

    Detre, S; King, N; Salter, J; MacLennan, K; McKinna, J A; Dowsett, M

    1994-01-01

    AIMS--(i) To assess the validity of an immunocytochemical technique for detecting pS2 protein in paraffin wax embedded tissue; (ii) to provide further data on the relation between pS2 protein and oestrogen receptor (ER) and progesterone receptor (PgR). METHODS--Breast cancer excision biopsy specimens were obtained from 35 previously untreated patients. An immunoradiometric assay was compared with an immunohistochemical method for measuring pS2 protein. ER and PgR were measured in cytosol fractions by enzyme immunoassay and the relation between the presence of these receptors and pS2 protein was assessed before and after subdivision of the women into groups of over or under 50 years of age. RESULTS--A good correlation was seen between the immunoradiometric and immunohistochemical methods for pS2 protein measurement (r = 0.84; p = 0.0001). Log-transformed data showed a significant correlation between increasing values of ER and pS2 protein (r = 0.45; p = 0.006) and to a lesser extent between pS2 protein and PgR (r = 0.38; p = 0.03). Correlations were also shown between pS2+ and PgR+ status (p = 0.01), and between ER and PgR positivity (p = 0.05; Fisher's exact test). pS2+ protein status was only associated with ER+ status in patients aged 50 years or less. CONCLUSIONS--The two methods for pS2 analysis are virtually interchangeable. This provides strong support for using immunohistochemistry for pS2 in paraffin wax embedded tissue. The association with ER positivity and pS2+ protein status only in the premenopausal patients may be due to the higher levels of oestrogenic stimuli in that group. Images PMID:8163696

  3. Primary breast cancer tumours contain high amounts of IgA1 immunoglobulin: an immunohistochemical analysis of a possible carrier of the tumour-associated Tn antigen.

    PubMed

    Welinder, Charlotte; Baldetorp, Bo; Blixt, Ola; Grabau, Dorthe; Jansson, Bo

    2013-01-01

    The Tn antigen (GalNAc alpha-O-Ser/Thr) as defined by the binding of the lectin, helix pomatia agglutinin (HPA) or anti-Tn monoclonal antibodies, is known to be exposed in a majority of cancers, and it has also been shown to correlate positively with the metastatic capacity in breast carcinoma. The short O-glycan that forms the antigen is carried by a number of different proteins. One potential carrier of the Tn antigen is immunoglobulin A1 (IgA1), which we surprisingly found in tumour cells of the invasive parts of primary breast carcinoma. Conventional immunohistochemical analysis of paraffin-embedded sections from primary breast cancers showed IgA1 to be present in the cytoplasm and plasma membrane of 35 out of 36 individual primary tumours. The immunohistochemical staining of HPA and anti-Tn antibody (GOD3-2C4) did to some extent overlap with the presence of IgA1 in the tumours, but differences were seen in the percentage of stained cells and in the staining pattern in the different breast cancers analysed. Anti-Tn antibody and HPA were also shown to specifically bind to a number of possible constellations of the Tn antigen in the hinge region of IgA1. Both reagents could also detect the presence of Tn positive IgA in serum. On average 51% of the tumour cells in the individual breast cancer tumour sections showed staining for IgA1. The overall amount of staining in the invasive part of the tumour with the anti Tn antibody was 67%, and 93% with HPA. The intra-expression or uptake of IgA1 in breast cancer makes it a new potential carrier of the tumour associated and immunogenic Tn antigen.

  4. Pituitary adenylate cyclase-activating peptide in the rat central nervous system: an immunohistochemical and in situ hybridization study.

    PubMed

    Hannibal, Jens

    2002-11-25

    In the present study the localization of pituitary adenylate cyclase-activating peptide (PACAP)-expressing cell bodies and PACAP projections were mapped in the adult rat brain and spinal cord by using immunohistochemistry and in situ hybridization histochemistry. A widespread occurrence of PACAP-containing cell bodies was found, with the greatest accumulation in several hypothalamic nuclei and in several brainstem nuclei, especially the habenular nuclei, the pontine nucleus, the lateral parabrachial nucleus (LPB), and the vagal complex. PACAP was also present in cell bodies in the olfactory areas, in neocortical areas, in the hippocampus, in the vestibulo- and cochlear nuclei, in cell bodies of the intermediolateral cell column of the spinal cord and in Purkinje cells of the cerebellum, in the subfornical organ, and in the organum vasculosum of the lamina terminalis. An intense accumulation of PACAP-immunoreactive (-IR) nerve fibers was observed throughout the hypothalamus, in the amydaloid and extended amygdaloid complex, in the anterior and paraventricular thalamic nuclei, in the intergeniculate leaflet, in the pretectum, and in several brainstem nuclei, such as the parabrachial nucleus, the sensory trigeminal nucleus, and the nucleus of the solitary tract. PACAP-IR nerve fibers were also found in the area postrema, the posterior pituitary and the choroid plexus, and the dorsal and ventral horn of the spinal cord. The widespread distribution of PACAP in the brain and spinal cord suggests that PACAP is involved in the control of many autonomic and sensory functions as well as higher cortical processes.

  5. Undifferentiated Carcinoma of the Endometrium: An Expanded Immunohistochemical Analysis Including PAX-8 and Basal-Like Carcinoma Surrogate Markers.

    PubMed

    Ramalingam, Preetha; Masand, Ramya P; Euscher, Elizabeth D; Malpica, Anais

    2016-09-01

    loss of this marker appears to be a more reliable discriminator than the loss of keratin expression in the differential diagnosis with endometrioid carcinoma or serous carcinoma. UCAe tends to be diffusely positive for p53, but patchy positive for p16. Although UCAe appears to share not only some histologic features with BLCB, but also some of its immunohistochemical features (loss of estrogen receptor, progesterone receptor, and Her-2/neu, a tendency to loose e-cadherin and to express CD44), UCAe appears not to be related to BLCB because it usually lacks the expression EGFR, CK5/6, and c-Kit. PMID:26598976

  6. Intravascular papillary endothelial hyperplasia: histomorphological and immunohistochemical features

    PubMed Central

    2013-01-01

    Background Intravascular papillary endothelial hyperplasia (IPEH) is a benign intravascular process with features mimicking other benign and malignant vascular proliferations. IPEH lesions predominate in the head-neck region and the extremities. The characteristic histomorphological feature of IPEH is a papillary structure covered with hyperplastic endothelial cells within the vascular lumen. It is critical that this clinically benign lesion should not be mistaken for well-differentiated vascular tumors. In addition to the characteristic histological features, other useful diagnostic features included the intra-luminal location of the lesion, an intimate association with the organizing thrombus, the absence of necrosis, cellular pleomorphism, and mitotic activity. In addition, immunohistochemistry may indicate the vascular origin and proliferative index. In this study, we evaluated histomorphological and immunohistochemical findings (CD31, CD34, FVIII, type IV collagen, SMA, MSA, CD105, and Ki-67 staining) of ten IPEH cases. Methods Ten IPEH cases were re-examined for a panel of histomorphological and immunohistochemical features. CD31, CD34, FVIII, Type IV collagen, SMA and MSA antibodies utilized for immunohistochemical analysis. The histomorphological and immunohistochemical findings were evaluated by two independent pathologists using light microscopy. Results All ten cases involved intraluminal lesions with characteristic features of IPEH. All ten cases (100%) were stained positive for CD31 and CD34. The degree of staining with FVIII, type IV collagen, SMA, and MSA was variable. Conclusion In this series of specimens, CD31 and CD34 were the most sensitive markers indicating the vascular origin of the lesion. Staining for the other vascular markers (FVIII, type IV collagen, SMA and MSA) was variable. Different maturation degrees of lesions may account for the variation in immunohistochemical staining. Few previous investigations evaluated a wide range of

  7. Ectopic hamartomatous thymoma: a clinicopathologic and immunohistochemical analysis of 21 cases with data supporting reclassification as a branchial anlage mixed tumor.

    PubMed

    Fetsch, John F; Laskin, William B; Michal, Michal; Remotti, Fabrizio; Heffner, Dennis; Ellis, Gary; Furlong, Mary; Miettinen, Markku

    2004-10-01

    This report describes the clinicopathologic and immunohistochemical findings in 21 cases of a highly distinctive tumor with a strong predilection for the lower neck region of adult males. Our study group consisted of 20 males and one female. The patients were 28 to 79 years old (mean age, 47 years; median age, 40 years), and they presented with solitary, lobular or multilobular masses ranging in size from 2.0 to 19.0 cm in greatest dimension (mean size, 5.1 cm; median, 4 cm). The tumors principally involved the lower neck region, usually in close proximity to the sternoclavicular joint. The preoperative duration of the lesions ranged from 2 months to 30 years. Histologically, the tumors were typically well marginated and composed of plump spindled cells, delicate spindled cells, mature adipose tissue, and epithelial cells, including both squamous and glandular elements. Epithelial-lined cysts were a focal finding in most cases and measured up to 2 cm in greatest dimension. Mitotic counts for the tumors ranged from 0 to 7 mitotic figures per 50 high power fields (mean mitotic count, 1.1 mitotic figures per 50 HPFs). Our immunohistochemical analysis revealed a complex immunophenotype with a diverse keratin profile. The plump spindled cells had a myoepithelial phenotype, as evidenced by the coexpression of keratins (5, 5/6, and 14), alpha-smooth muscle actin, CD10, and to a lesser extent, calponin. No compelling evidence for thymic differentiation was observed. The patients were initially managed by biopsy or partial resection (n = 4), simple local excision (n = 16), or an unspecified procedure (n = 1). Clinical follow-up of > or =3 years was available for 10 patients (48%). Two patients had recurrent disease, but there were no metastases or tumor-related deaths. A derivation from sequestered branchial epithelium is likely, but evidence for a thymic component is tenuous, at best. Our data support reclassification of this distinctive process as a branchial anlage mixed

  8. Immunohistochemical analysis of the Akt/mTOR/4E-BP1 signalling pathway in canine haemangiomas and haemangiosarcomas.

    PubMed

    Murai, A; Abou Asa, S; Kodama, A; Sakai, H; Hirata, A; Yanai, T

    2012-11-01

    The specific signalling pathways that are deregulated in canine endothelial tumours have not yet fully elucidated. Therefore, the aim of the present study was to examine activation of the Akt/mammalian target of rapamycin (mTOR)/eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) signalling pathway in spontaneously arising canine haemangiomas (HAs) and haemangiosarcomas (HSAs) in order to identify novel molecular targets for treatment. Surgically-resected samples of HA (n = 27), HSA (n = 37), granulation tissue (n = 4) and normal skin (n = 4) were investigated by immunohistochemistry. Approximately 80% of the HSA samples had moderate to intense expression of phosphorylated Akt at Ser473 (p-Akt Ser473), p-Akt Thr308, p-4E-BP1 Thr37/46 and eukaryotic initiation factor 4E, which was significantly higher than in the HAs and was similar to the expression in activated endothelial cells (ECs). Although p-mTOR complex1 (p-mTORC1) Ser2448 was expressed by most of the activated ECs, only 35% of the HSA samples had weak to moderate expression. Because mTORC2 and phosphorylates Akt Ser473 was activated in HSA samples, the present findings suggest that the mTORC2/Akt/4E-BP1 pathway, regulated independently of mTORC1, may be important for targeting therapy in canine HSAs. PMID:22789858

  9. Expression of c-kit, a proto-oncogene of the murine W locus, in cerebella of normal and neurological mutant mice: immunohistochemical and in situ hybridization analysis.

    PubMed

    Takeda, H; Yoshiki, A; Nishikawa, S; Nishikawa, S; Kunisada, T; Sakakura, T; Amanuma, H; Kusakabe, M

    1992-10-01

    The c-kit proto-oncogene encodes a receptor tyrosine kinase and is allelic with the murine white-spoting (W) locus. Although no apparent defects in the brain have been reported in W mutant mice, brain tissue, especially cerebellum, shows a high level of c-kit transcription. In the present study, sites of c-kit expression in the cerebellum were exained by immunohistochemical and in situ hybridization techniques. Immunohistochemistry with a monoclonal antibody against c-Kit protein revealed that the c-Kit protein was localized close to the Purkinje cell soma in the region facing the granular cell layer. Similar distribution of the c-Kit protein was observed in cerebella of mutant mice in which the Purkinje cell (pcd) or the granular cell layer (weaver) is missing. These data suggest that the c-Kit protein is produced not by the Purkinje cell nor by the granular cell but by the cells present in the molecular layer and that the protein is then transported to the region around the Purkinje cell soma. This interpretation was supported by in situ hybridization analysis: cells containing the c-kit transcripts were found only in the molecular layer, while the granular and Purkinje cells were negative.

  10. Immunohistochemical Determination of p53 Protein Overexpression for Predicting p53 Gene Mutations in Hepatocellular Carcinoma: A Meta-Analysis

    PubMed Central

    Deng, Miao; Liu, Dechun; Ma, Qingyong; Feng, Xiaoshan

    2016-01-01

    Background Whether increased expression of the tumor suppressor protein p53 indicates a p53 gene mutation in hepatocellular carcinoma (HCC) remains unclear. We conducted a meta-analysis to determine whether p53 protein overexpression detected by immunohistochemistry (IHC) offers a diagnostic prediction for p53 gene mutations in HCC patients. Methods Systematic literature searches were conducted with an end date of December 2015. A meta-analysis was performed to estimate the diagnostic accuracy of IHC-determined p53 protein overexpression in the prediction of p53 gene mutations in HCC. Sensitivity, subgroup, and publication bias analyses were also conducted. Results Thirty-six studies were included in the meta-analysis. The results showed that the overall sensitivity and specificity for IHC-determined p53 overexpression in the diagnostic prediction of p53 mutations in HCC were 0.83 (95% CI: 0.80–0.86) and 0.74 (95% CI: 0.71–0.76), respectively. The summary positive likelihood ratio (PLR) and negative likelihood ratio (NLR) were 2.65 (95% CI: 2.21–3.18) and 0.36 (95% CI: 0.26–0.50), respectively. The diagnostic odds ratio (DOR) of IHC-determined p53 overexpression in predicting p53 mutations ranged from 0.56 to 105.00 (pooled, 9.77; 95% CI: 6.35–15.02), with significant heterogeneity between the included studies (I2 = 40.7%, P = 0.0067). Moreover, subgroup and sensitivity analyses did not alter the results of the meta-analysis. However, potential publication bias was present in the current meta-analysis. Conclusion The upregulation of the tumor suppressor protein p53 was indeed linked to p53 gene mutations. IHC determination of p53 overexpression can predict p53 gene mutations in HCC patients. PMID:27428001

  11. Podoplanin expression in tumor-free resection margins of oral squamous cell carcinomas: an immunohistochemical and fractal analysis study.

    PubMed

    Margaritescu, C; Raica, M; Pirici, D; Simionescu, C; Mogoanta, L; Stinga, A C; Stinga, A S; Ribatti, D

    2010-06-01

    Podoplanin is involved in tumorigenesis and cancer progression in head and neck malignancies and its expression is not restricted to lymphatic vessel endothelium. The aim of this study was to establish podoplanin expression in the tumor-free resection margins of oral squamous cell carcinomas (OSCCs) and to evaluate the geometric complexity of the lymphatic vessels in oral mucosa by utilizing fractal analysis. As concerns the podoplanin expression in noncancerous tissue, forty tumor-free resection margins from OSCCs were investigated utilizing immunohistochemistry for D2-40 antibody and image densitometry analysis. Podoplanin expression was extremely low in basal cells, especially in resection margins of OSCCs developed in the lower lip regions. However, a highly variable D2-40 expression in tumor-free resection margins associated with hyperplastic or dysplastic lesions was identified. Moreover, podoplanin expression also extended to the basal layer of the lower lip skin appendages, the myoepithelial cells of acini and ducts of minor salivary glands, and other structures from the oral cavity. As concerns the study of the density and complexity of oral lymphatic vessels architecture by means of immunohistochemistry (D2-40, CD31 and Ki-67 antibodies) and fractal analysis, we demonstrated that in normal oral mucosa the geometry of the lymphatic vessels was less complex at the level of the lower lip compared to the anterior part of the oral floor mucosa or the tongue. A comparative analysis between the normal and pathological aspects revealed statistically significant differences between the fractal dimension (FD) of the vessels' outline, especially in the tongue. Fractal analysis proved an increasing lymphatic network complexity from normal to premalignant oral mucosal lesions, providing additional prognostic information in oral malignant tumors. PMID:20376776

  12. Activation of coagulation and angiogenesis in cancer: immunohistochemical localization in situ of clotting proteins and vascular endothelial growth factor in human cancer.

    PubMed Central

    Shoji, M.; Hancock, W. W.; Abe, K.; Micko, C.; Casper, K. A.; Baine, R. M.; Wilcox, J. N.; Danave, I.; Dillehay, D. L.; Matthews, E.; Contrino, J.; Morrissey, J. H.; Gordon, S.; Edgington, T. S.; Kudryk, B.; Kreutzer, D. L.; Rickles, F. R.

    1998-01-01

    Thrombin-catalyzed, cross-linked fibrin (XLF) formation is a characteristic histopathological finding in many human and experimental tumors and is thought to be of importance in the local host defense response. Although the pathogenesis of tumor-associated fibrin deposition is not entirely clear, several tumor procoagulants have been described as likely primary stimuli for the generation of thrombin (and XLF) in the tumor microenvironment (TME). In a previous study of a variety of human tumors we have shown that tissue factor (TF) is the major procoagulant. However, the relative contribution to fibrin deposition in the TME of tumor cell TF and host cell TF (eg, macrophage-derived) was not established. In addition, recent evidence has implicated TF in the regulation of the synthesis of the pro-angiogenic factor vascular endothelial growth factor (VEGF) by tumor cells. In the current study we used in situ techniques to determine the cellular localization of XLF, TF, VEGF, and an alternative tumor procoagulant, so-called cancer procoagulant (CP), a cysteine protease that activates clotting factor X. In lung cancer we have found XLF localized predominantly to the surface of tumor-associated macrophages, as well as to some endothelial cells and perivascular fibroblasts in the stromal area of the tumors co-distributed with TF at the interface of the tumor and host cells. Cancer pro-coagulant was localized to tumor cells in several cases but not in conjunction with the deposition of XLF. TF and VEGF were co-localized in both lung cancer and breast cancer cells by in situ hybridization and immunohistochemical staining. Furthermore, a strong relationship was found between the synthesis of TF and VEGF levels in human breast cancer cell lines (r2 = 0.84; P < 0.0001). Taken together, these data are consistent with a highly complex interaction between tumor cells, macrophages, and endothelial cells in the TME leading to fibrin formation and tumor angiogenesis. Images Figure 1

  13. Neurofilament protein defines regional patterns of cortical organization in the macaque monkey visual system: a quantitative immunohistochemical analysis

    NASA Technical Reports Server (NTRS)

    Hof, P. R.; Morrison, J. H.; Bloom, F. E. (Principal Investigator)

    1995-01-01

    Visual function in monkeys is subserved at the cortical level by a large number of areas defined by their specific physiological properties and connectivity patterns. For most of these cortical fields, a precise index of their degree of anatomical specialization has not yet been defined, although many regional patterns have been described using Nissl or myelin stains. In the present study, an attempt has been made to elucidate the regional characteristics, and to varying degrees boundaries, of several visual cortical areas in the macaque monkey using an antibody to neurofilament protein (SMI32). This antibody labels a subset of pyramidal neurons with highly specific regional and laminar distribution patterns in the cerebral cortex. Based on the staining patterns and regional quantitative analysis, as many as 28 cortical fields were reliably identified. Each field had a homogeneous distribution of labeled neurons, except area V1, where increases in layer IVB cell and in Meynert cell counts paralleled the increase in the degree of eccentricity in the visual field representation. Within the occipitotemporal pathway, areas V3 and V4 and fields in the inferior temporal cortex were characterized by a distinct population of neurofilament-rich neurons in layers II-IIIa, whereas areas located in the parietal cortex and part of the occipitoparietal pathway had a consistent population of large labeled neurons in layer Va. The mediotemporal areas MT and MST displayed a distinct population of densely labeled neurons in layer VI. Quantitative analysis of the laminar distribution of the labeled neurons demonstrated that the visual cortical areas could be grouped in four hierarchical levels based on the ratio of neuron counts between infragranular and supragranular layers, with the first (areas V1, V2, V3, and V3A) and third (temporal and parietal regions) levels characterized by low ratios and the second (areas MT, MST, and V4) and fourth (frontal regions) levels characterized by

  14. Composite pheochromocytoma/ganglioneuroma of the adrenal gland associated with multiple endocrine neoplasia 2A: case report with immunohistochemical analysis.

    PubMed

    Brady, S; Lechan, R M; Schwaitzberg, S D; Dayal, Y; Ziar, J; Tischler, A S

    1997-01-01

    We report a case of composite pheochromocytoma/ganglioneuroma arising in a background of diffuse and nodular medullary hyperplasia in the adrenal gland of a 34-year-old man with multiple endocrine neoplasia 2a (MEN 2a). Cells were histologically classified as chromaffin or chromaffin-like (small typical-appearing pheochromocytoma cells), neuron-like (possessing ganglion cell morphology), and intermediate. We speculate that these cell types may represent a spectrum of differentiation of a neoplastic clone, with the intermediate cells representing a transitional stage between chromaffin cells and neurons. All three cell types in the composite tumor and all chromaffin cells in both nodular and nonnodular areas of the remaining medulla were strongly immunoreactive for tyrosine hydroxylase, the rate-limiting enzyme in catecholamine synthesis. In contrast, neuron-like cells (and to a variable extent intermediate cells) displayed selective loss of expression of phenylethanolamine-N-methyltransferase (PNMT), the enzyme that synthesizes epinephrine. Proliferative activity of the composite tumor and both the nodular and nonnodular medulla was studied by staining for the endogenous cell proliferation antigen Ki-67, using monoclonal antibody MIB-1. MIB-1 labeling was highest in Schwann cell areas of the composite tumor, followed by chromaffin-like cells in the composite tumor and in the separate nodules. Labeling was absent in neuron-like cells, consistent with the cells' postulated status as terminally differentiated derivatives of a chromaffin cell precursor, and was highly variable in nonnodular areas of the medulla. The latter observation suggests topographical variation in signals that drive chromaffin cell proliferation in MEN.

  15. Immunohistochemical and morphometric analysis of intestinal full-thickness biopsy samples from cats with lymphoplasmacytic inflammatory bowel disease.

    PubMed

    Marsilio, S; Kleinschmidt, S; Nolte, I; Hewicker-Trautwein, M

    2014-05-01

    The distribution and numbers of CD3(+) T lymphocytes, immunoglobulin(+) plasma cells and calprotectin (L1)(+) macrophages was analyzed in full-thickness, formalin-fixed biopsy samples from the small intestine (duodenum, jejunum and ileum) and from the colon from nine cats with clinical signs of inflammatory bowel disease (IBD). All animals had lymphoplasmacytic enteritis or lymphoplasmacytic enterocolitis. Equivalent samples from the same intestinal regions from 12 healthy pet cats served as controls. Labelled cells in the lamina propria were counted by computer-aided morphometry. The different cell types were similarly distributed in both groups, but there were differences in their numbers. There were more CD3(+) T cells in the duodenum and jejunum of cats with IBD; however, the difference was only significant for the duodenum. There were significantly more IgA(+) cells in the duodenal crypt region. There were significantly more IgG(+) cells in the lower jejunal crypt region. Plasma cells expressing IgM were decreased in cats with IBD, but the difference was not significant. L1(+) macrophages were significantly decreased in the lower crypt area of the colon in cats with IBD and there was a trend to decreased L1(+) cells in the upper crypt area of the duodenum and jejunum. Comparison of the results of this study with previous findings on endoscopically-obtained duodenal biopsy samples from cats with IBD revealed some differences. These discrepancies might relate to differences between control cat populations, types of biopsy samples, methodological factors such as different counting techniques and the activity of the disease at the time of sampling.

  16. Significantly greater expression of ER, PR, and ECAD in advanced-stage low-grade ovarian serous carcinoma as revealed by immunohistochemical analysis.

    PubMed

    Wong, Kwong-Kwok; Lu, Karen H; Malpica, Anais; Bodurka, Diane C; Shvartsman, Hyun S; Schmandt, Rosemarie E; Thornton, Angela D; Deavers, Michael T; Silva, Elvio G; Gershenson, David M

    2007-10-01

    A 2-tier system that classifies ovarian serous carcinoma (OSC) as low grade or high grade is gaining acceptance. Women with low-grade OSC generally have higher 5-year survival rates than do women with high-grade OSC. We examined the expression of various markers to further understand the molecular differences between low-grade and high-grade OSCs: the potential therapeutic targets or prognostic markers Her-2/neu, estrogen receptor, and progesterone receptor (PR); the metastasis-associated markers cyclin D1 (BCL1), E-cadherin, matrix metalloproteinase (MMP) 2, and MMP-9; and the cell proliferation-associated markers BCL1, Ki-67 antigen (Ki-67), and p53. For this immunohistochemical analysis, we used paraffin-embedded specimens from 47 patients with advanced-stage low-grade OSC and from 49 patients with advanced-stage high-grade OSC. Our results showed that low-grade tumors expressed significantly higher levels of estrogen receptor, PR, and E-cadherin than did high-grade tumors, suggesting the involvement of gonadal steroid hormones, especially in the pathogenesis of low-grade OSC; the PR positivity was also observed in the stromal component of these low-grade tumors. On the other hand, high-grade tumors trended toward increased expression of MMP-9, BCL1, p53, and Ki-67, and robust MMP-9 positivity was observed in the stromal component of these high-grade tumors. These differences may lead to the development of different therapeutic strategies for women with either the low-grade or the high-grade form of OSC.

  17. Assessment of Epidermal Growth Factor Receptor (EGFR) signaling in paired colorectal cancer and normal colon tissue samples using computer-aided immunohistochemical analysis.

    PubMed

    Messersmith, Wells; Oppenheimer, Darin; Peralba, Josep; Sebastiani, Valeria; Amador, Maria; Jimeno, Antonio; Embuscado, Erlinda; Hidalgo, Manuel; Iacobuzio-Donahue, Christine

    2005-12-01

    The Epidermal Growth Factor Receptor (EGFR) plays a role in multiple tumor cell processes and is targeted by several anticancer therapies. Although EGFR mutations may determine tumor susceptibility in a small proportion of patients, knowledge of the EGFR signaling pathway status in tumors may help guide further drug development and hypothesis-driven combination studies. We aimed to validate and apply a novel computer-aided immunohistochemical (IHC) technique to characterize the status of EGFR signaling in matched colorectal tumor and normal colon tissue samples. Tissue Microarrays (TMA)were made from both cancerous and normal colorectal tissue in 18 patients and stained with antibodies against EGFR, phospho-EGFR (pEGFR), Akt, pAkt, MAPK, and pMAPK. TMA's were quantitatively scored using the Automated Cellular Imaging System (ACIS II, Chromavision, Inc). ACIS was compared against cell line Western blotting, ELISA, and visual scoring (0-3+) by a pathologist. We found that ACIS analysis was highly reproducible and results were well correlated with other techniques. A post-scan "image microdissection" technique of analyzing heterogeneous human samples showed good correlation between paired human samples [Pearson correlation for tumors, 0.922 (p < .001)]. Cancer samples had markedly higher staining of pEGFR, Akt, pAkt, MAPK, and pMAPK. We conclude that ACIS IHC of human tissue samples is quantitative, reproducible, and correlates with Western blots and ELISA in cell line pellets as well as pathologist's scores of human samples. Colorectal tumors show higher staining of pEGFR and downstream effectors compared to matched normal colorectal tissues.

  18. IMMUNOHISTOCHEMICAL DIFFERENTIATION OF TRIPLE NEGATIVE BREAST CANCER.

    PubMed

    Lesar, Miroslav; Stanec, Mladen; Lesar, Nikola; Vrdoljak, Danko Velimir; Zore, Zvonimir; Banović, Marija; Brozović, Gordana

    2016-03-01

    Based on immunohistochemical staining for the basal markers cytokeratin 5/6 (CK 5/6), cytokeratin 14 (CK 14) and P-cadherin, triple negative tumors (TNT) are divided into two groups: 1) basal-like (BL) positive for one or all three markers; and 2) non basal-like (NBL) negative for all three markers. Even though the different origin of the cells of these two types of tumors implies different biological properties, they had been treated as one entity until recently. This paper analyzes TNT collected from 150 patients and distributed into two groups according to the results of immunohistochemical analysis, i.e. BL 116 (77.3%) and NBL 34 (22.67%). In this study, CK 5/6, CK 14 and P-cadherin were used as markers for identifying BL tumors. The immunohistochemical reaction was positive for CK 5/6 in 37%, for CK 14 in 50.86% and for P-cadherin in 68.34% of cases. The subclassification of triple negative breast cancer using the basal markers CK 5/6, CK 14 and P-cadherin has enabled identification of BL and NBL breast cancers in a proportion that is in line with the only accurate analysis of TNT gene expression. Using the mentioned combination of markers in daily practice is easy to perform and economically affordable. PMID:27333711

  19. Canalicular adenoma--search for the cell of origin: ultrastructural and immunohistochemical analysis of 7 cases and review of the literature.

    PubMed

    Huebner, Thomas A; Almubarak, Hussain; Drachenberg, Cinthia B; Papadimitriou, John C

    2014-04-01

    Canalicular adenoma (CA) is a rare, benign epithelial neoplasm of the salivary glands. Historically considered to be a variant of basal cell adenoma, this "monomorphic" adenoma has a distinct clinical, morphologic, and immunohistochemical profile. The putative cell of origin remains a topic of debate. A combination of morphology, immunohistochemistry, and ultrastructural analyses have been employed to determine histogenesis, but the interpretations of these studies have implicated multiple different cell types along the salivary gland duct as the cell of origin. The authors sought to further characterize CA using electron microscopy, immunohistochemistry, and special and immuno-stains on 7 cases. Their morphologic, immunohistochemical, and ultrastructural findings support a cell of origin demonstrating features of both the intercalated duct cells and the striated duct luminal epithelial cells.

  20. Solitary fibrous tumour: clinicopathological, immunohistochemical, and ultrastructural analysis of 12 cases arising in soft tissues, nasal cavity and nasopharynx, urinary bladder and prostate.

    PubMed

    Mentzel, T; Bainbridge, T C; Katenkamp, D

    1997-06-01

    The clinicopathological features of 12 extraserosal solitary fibrous tumours (SFT) are described. The age of the patients ranged from 18 to 72 years (mean: 48.2 years; median: 54 years); 5 were female patients. Seven lesions arose in soft tissue (5 in perifascial, and 1 each in subcutaneous and intramuscular tissues). They were situated in the groin (2 cases) and the neck, right buttock, left scapula, upper arm, and anterior abdominal wall (1 case each). One polypoid lesion was seen in in the nasal cavity and 1 in the nasopharynx; 2 neoplasms arose in the urinary bladder and 1 was located in the prostate and periprostatic tissue. Nine lesions were excised; in 1 patient wide excision was performed and in 2 patients, transurethral resection. Limited follow-up of 3 cases revealed a benign clinical course. The size of the neoplasms ranged from 1.7 cm to 20.0 cm (mean: 5.4 cm; median: 3.5 cm). Histologically, the neoplasms were well circumscribed and composed of cytologically bland spindle cells arranged without an obvious pattern; focally storiform or fascicular growth patterns were seen. Tumour cells were separated by thick bands of collagen demonstrating foci of keloid-like hyalinization. Prominent vascularity showing a haemangiopericytoma-like vascular pattern and vessels with thick, hyalinized vessel walls were seen in all cases. Increased mitotic activity was noted in 2 soft tissue cases (4-6 mitoses in 10 high-power fields); the other cases showed fewer than 2 mitotic figures in 10 highpower fields. Immunohistochemically, all cases tested stained positively for vimentin, CD34 and CD99, and 2 cases showed focal myofibroblastic differentiation. Two cases examined ultrastructurally showed a fibroblastic phenotype; focally pinocytic vesicles and microfilaments were identified. SFT represents a distinct neoplasm that should be included in the differential diagnosis of spindle-cell neoplasms in soft tissue, nasal cavity and nasopharynx, urinary bladder, and prostate

  1. Determination of HER2 and p53 Mutations by Sequence Analysis Method and EGFR/Chromosome 7 Gene Status by Fluorescence in Situ Hybridization for the Predilection of Targeted Therapy Modalities in Immunohistochemically Triple Negative Breast Carcinomas in Turkish Population.

    PubMed

    Pala, Emel Ebru; Bayol, Umit; Keskin, Elif Usturali; Ozguzer, Alp; Kucuk, Ulku; Ozer, Ozge; Koc, Altug

    2015-09-01

    Triple negative breast cancer (TNBC), an agressive subtype accounts nearly 15 % of all breast carcinomas. Conventional chemotherapy is the only treatment modality thus new, effective targeted therapy methods have been investigated. Epidermal growth factor receptor (EGFR) inhibitors give hope according to the recent studies results. Also therapeutic agents have been tried against aberrant p53 signal activity as TNBC show high p53 mutation rates. Our aim was to detect the incidence of mutations/amplifications identified in TNBC in our population. Here we used sequence analysis to detect HER2 (exon 18-23), p53 (exon 5-8) mutations; fluorescence in situ hybridization (FISH) method to analyse EGFR/chromosome 7 centromere gene status in 82 immunohistochemically TNBC. Basaloid phenotype was identified in 49 (59.8 %) patients. EGFR amplification was noted in 5 cases (6.1 %). All EGFR amplified cases showed EGFR overexpression by immunohistochemistry (IHC). p53 mutations were identified in 33 (40.2 %) cases. Almost 60 % of the basal like breast cancer cases showed p53 mutation. Only one case showed HER2 mutation (exon 20:g.36830_3). Our results showed that gene amplification is not the unique mechanism in EGFR overexpression. IHC might be used in the decision of anti-EGFR therapy in routine practice. p53 mutation rate was lower than the rates reported in the literature probably due to ethnic differences and low sensitivity of sanger sequences in general mutation screening. We also established the rarity of HER2 mutation in TNBC. In conclusion EGFR and p53 are the major targets in TNBC also for our population.

  2. Immunohistochemical Expression of Estrogen and Progesterone Receptors in Human Colorectal Adenoma and Carcinoma Using Specified Automated Cellular Image Analysis System: A Clinicopathological Study

    PubMed Central

    Qasim, Ban Jumaa; Ali, Hussam Hasson; Hussein, Alaa Ghani

    2011-01-01

    Objectives To evaluate the immunohistochemical expression of estrogen receptors (ER) and progesterone receptors (PR) in colorectal adenoma and adenocarcinoma and to correlate this immunohistochemical expression with different clinicopathological parameters. Methods The study was retrospectively designed. A total of 86 tissue samples, including 33 paraffin blocks from patients with colorectal adenomas, 33 paraffin blocks from patients with colorectal adenocarcinomas and a control group of 20 samples of non-tumorous colonic tissue, were included in the study. Results The frequency of expression of ER and PR showed a gradual increase from control through adenoma to carcinoma. The frequencies of expression of ER in the control, adenoma and carcinoma were (10%, 15.15% and 42.42% respectively, p<0.001), while the frequency of expression for PR were (10%, 24.24% and 36.36% respectively, p<0.001). Strong ER and PR staining was mainly seen in carcinoma cases (42.42%, 36.36%, respectively) in comparison with adenoma (9.09%, 15.15%, respectively) and control (0%, 0%, respectively). The three digital parameters of ER and PR immunohistochemical expression (Area [A], Number of objects [N], and intensity [I]) were significantly increased in a sequence of normal mucosa-adenoma-carcinoma. There was a significant positive correlation between ER and PR in adenoma (r=0.312, p=0.034) and carcinoma (r=0.321, p=0.0398). Conclusion ER and PR expression increased in a sequence of; normal colonic mucosa-adenoma-carcinoma, and a positive correlation was observed between ER and PR expression in colonic adenoma and carcinoma specimen indicating that ER and PR may play a role in colorectal carcinogenesis. PMID:22125723

  3. Immunohistochemical analysis of the skin in junctional epidermolysis bullosa using laminin 5 chain specific antibodies is of limited value in predicting the underlying gene mutation.

    PubMed

    McMillan, J R; McGrath, J A; Pulkkinen, L; Kon, A; Burgeson, R E; Ortonne, J P; Meneguzzi, G; Uitto, J; Eady, R A

    1997-06-01

    The anchoring filament protein laminin 5 is composed of three polypeptide chains (alpha 3, beta 3 and gamma 2) each encoded by separate genes (LAMA3, LAMB3 and LAMC2, respectively). Mutations in any of these three genes may give rise to the autosomal recessive blistering skin disease, junctional epidermolysis bullosa. At present, there is no easy way of predicting which of these three genes might harbour the pathogenetic laminin 5 mutations in a case of junctional epidermolysis bullosa. In this study, we assessed whether immunohistochemistry might be helpful in this regard. We performed immunohistochemical labelling of the dermal-epidermal junction using alpha 3, beta 3 and gamma 2 chain-specific antibodies in 11 patients with junctional epidermolysis bullosa, in whom the laminin 5 mutations had been previously delineated. Although, labelling for the laminin 5 chain bearing the mutations was attenuated or undetectable in all cases, a complete absence of labelling or a reduction in the staining intensity for the other two chains was also seen in all cases. The results showed that immunohistochemical labelling of the dermal-epidermal junction using alpha 3, beta 3 and gamma 2 chain-specific antibodies is not a specific indicator for which of the laminin 5 chain genes contains the pathogenetic mutations, and is therefore unreliable in screening for individual laminin 5 gene mutations in cases of junctional epidermolysis bullosa. PMID:9217810

  4. Neutron activation analysis system

    DOEpatents

    Taylor, M.C.; Rhodes, J.R.

    1973-12-25

    A neutron activation analysis system for monitoring a generally fluid media, such as slurries, solutions, and fluidized powders, including two separate conduit loops for circulating fluid samples within the range of radiation sources and detectors is described. Associated with the first loop is a neutron source that emits s high flux of slow and thermal neutrons. The second loop employs a fast neutron source, the flux from which is substantially free of thermal neutrons. Adjacent to both loops are gamma counters for spectrographic determination of the fluid constituents. Other gsmma sources and detectors are arranged across a portion of each loop for deterMining the fluid density. (Official Gazette)

  5. An analysis of the sensitivity and specificity of MHC-I and MHC-II immunohistochemical staining in muscle biopsies for the diagnosis of inflammatory myopathies.

    PubMed

    Rodríguez Cruz, Pedro M; Luo, Yue-Bei; Miller, James; Junckerstorff, Reimar C; Mastaglia, Frank L; Fabian, Victoria

    2014-12-01

    Although there have been several previous reports of immunohistochemical staining for MHC antigens in muscle biopsies, there appears to be a lack of consensus about its routine use in the diagnostic evaluation of biopsies from patients with suspected inflammatory myopathy. Positive MHC-I staining is nonspecific but is widely used as a marker for inflammatory myopathy, whilst the role of MHC-II staining is not clearly defined. We investigated the sensitivity and specificity of MHC-I and MHC-II immunostaining for the diagnosis of inflammatory myopathy in a large group of biopsies from a single reference laboratory. Positive staining for MHC-I was found to have a high sensitivity in biopsies from patients with inflammatory myopathy but a very low specificity, as it was also common in other non-inflammatory myopathies and neurogenic disorders. On the other hand, MHC-II positivity had a much higher specificity in all major subgroups of inflammatory myopathy, especially inclusion body myositis. The findings indicate that the combination of MHC-I and MHC-II staining results in a higher degree of specificity for the diagnosis of inflammatory myopathy and that in biopsies with inflammation, positive MHC-II staining strongly supports the diagnosis of an immune-mediated myopathy. We recommend that immunohistochemical staining for both MHC-I and MHC-II should be included routinely in the diagnostic evaluation of muscle biopsies from patients with suspected inflammatory myopathy. However, as the sensitivity and interpretation of MHC staining may depend on the technique used, further studies are needed to compare procedures in different centres and develop standardised protocols.

  6. Rheumatoid lymphadenopathy: a morphological and immunohistochemical study.

    PubMed Central

    Kondratowicz, G M; Symmons, D P; Bacon, P A; Mageed, R A; Jones, E L

    1990-01-01

    Sixteen lymph nodes from 14 patients with rheumatoid arthritis were examined immunohistochemically and morphometrically and compared with 10 control nodes showing follicular hyperplasia from patients without rheumatoid disease. Frozen material was available from nine patients and all controls. The nodes from patients with rheumatoid arthritis seemed to share characteristic features. The most striking of these was follicular hyperplasia in which the germinal centres, in spite of being quite large, showed relatively sparse proliferative activity. The nodes often showed infiltration of germinal centres by CD8 positive T lymphocytes and contained fewer IL2R positive cells in the paracortex than controls. These and other features may have some correlation with disease activity. Lymphadenopathy in rheumatoid arthritis may not just be a manifestation of joint inflammation but an active component of this multisystem disease and may reflect a widespread immunological abnormality. Images PMID:2318986

  7. Experiment K-6-22. Growth hormone regulation, synthesis and secretion in microgravity. Part 1: Somatotroph physiology. Part 2: Immunohistochemical analysis of hypothalamic hormones. Part 3: Plasma analysis

    NASA Technical Reports Server (NTRS)

    Grindeland, R.; Vale, W.; Hymer, W.; Sawchenko, P.; Vasques, M.; Krasnov, I.; Kaplanski, A.; Victorov, I.

    1990-01-01

    The objectives of the 1887 mission were: (1) to determine if the results of the SL-3 pituitary gland experiment (1) were repeatable; and (2) to determine what effect a longer mission would have on the rat pituitary gland growth hormone (GH) system. In the 1887 experiment two issues were considered especially important. First, it was recognized that cells prepared from individual rat pituitary glands should be considered separately so that the data from the 5 glands could be analyzed in a statistically meaningful way. Second, results of the SL-3 flight involving the hollow fiber implant and HPLC GH-variant experiments suggested that the biological activity of the hormone had been negatively affected by flight. The results of the 1887 experiment documented the wisdom of addressing both issues in the protocol. Thus, the reduction in secretory capacity of flight cells during subsequent extended cell culture on Earth was documented statistically, and thereby established the validity of the SL-3 result. The results of both flight experiments thus support the contention that there is a secretory lesion in pituitary GH cells of flight animals. The primary objective of both missions was a clear definition of the effect of spaceflight on the GH cell system. There can no longer be any reasonable doubt that this system is affected in microgravity. One explanation for the reason(s) underlying the better known effects of spaceflight on organisms, viz. changes in bone, muscle and immune systems may very well rest with such changes in bGH. In spite of the fact that rats in the Cosmos 1887 flight were on Earth for two days after flight, the data show that the GH system had still not recovered from the effects of flight. Many questions remain. One of the more important concerns the GRF responsiveness of somatotrophs after flight. This will be tested in an upcoming experiment.

  8. Comparative immunohistochemical analysis of the expression of cytokeratins, vimentin and alpha-smooth muscle actin in human foetal mesonephros and metanephros.

    PubMed

    Magro, G; Perris, R; Romeo, R; Marcello, M; Lopes, M; Vasquez, E; Grasso, S

    2001-04-01

    The human mesonephros is currently regarded as a simplified version of the foetal metanephros, primarily due to the close morphological resemblance between these two structures. The aim of the present study was to define whether human mesonephric and foetal metanephric nephrons share immunophenotypical traits in their corresponding structures (glomeruli, proximal and distal tubules). For this purpose we first investigated immunohistochemically the overall expression and topographical distribution of cytokeratins 7, 8, 18, 19, and 20, vimentin and alpha-smooth muscle actin in mature mesonephric nephrons and compared the results with those obtained in maturing-stage foetal metanephric nephrons. No expression of cytokeratins 7 and 20 was found. Cytokeratins 8, 18, and 19 and vimentin showed a restricted and basically coincident expression along the different components of both mesonephric and metanephric nephrons. These findings indicate that the intermediate filament protein profile of human mature mesonephric nephrons closely recapitulates that observed in developing metanephros and thereby strengthens the concept that human mesonephros, a transient ontogenic structure, is largely similar to the foetal metanephros. The sole difference between human mesonephros and foetal metanephros was the divergent expression of alpha-smooth muscle actin. This protein exhibited an increasingly accentuated mesangial expression paralleling the morphological maturation of metanephric glomerulus, whereas it was absent from the mesonephric one. This would suggest that the mesangial cells in these two renal structures have a different function during the foetal life.

  9. Kinetics of UV light-induced cyclobutane pyrimidine dimers in human skin in vivo: an immunohistochemical analysis of both epidermis and dermis.

    PubMed

    Katiyar, S K; Matsui, M S; Mukhtar, H

    2000-12-01

    It is well known that UV exposure of human skin induces DNA damage, and the cumulative effect of such repeated damage is an important contributor to the development of skin cancer. Here, we demonstrate UV dose- and time-dependent induction of DNA damage in the form of cyclobutane pyrimidine dimers (CPD) in skin cells following a single exposure of human skin to UV radiation. CPD+ cells were identified by an immunohistochemical technique using monoclonal antibodies to thymine dimers. The percentage of CPD+ cells was UV dose-dependent, even a suberythemal (0.5 minimal erythemal dose [MED]) dose resulted in detectable level of cells that contained pyrimidine dimers. Forty-eight hours after irradiation the percent of total epidermal cells positive for CPD ranged from 19 +/- 8, 36 +/- 10, 57 +/- 12 and 80 +/- 10, and total percent dermal cells positive for CPD ranged from 1 +/- 1, 7 +/- 3, 16 +/- 3 and 20 +/- 5, respectively, following 0.5, 1.0, 2.0 and 4.0 MED. CPD were also observed in deeper reticular dermis, which suggest the penetrating ability of UV radiation into the skin. The change in CPD+ cells from 0.5 to 240 h post-UV exposure in both epidermal and dermal compartments of the skin was also quantitated. CPD+ cells were observed in skin biopsies at early time points after UV exposure which remained elevated for 48 h, then declined significantly by 3 days post-UV. A close examination of the skin at and after 3 days following UV exposure indicates the significant removal of DNA damaged cells from the epidermis. Ten days after UV exposure the levels of CPD+ cells in both epidermis and dermis were not significantly different from that in unirradiated skin.

  10. Clear Cell Renal Cell Carcinoma With Borderline Features of Clear Cell Papillary Renal Cell Carcinoma: Combined Morphologic, Immunohistochemical, and Cytogenetic Analysis.

    PubMed

    Williamson, Sean R; Gupta, Nilesh S; Eble, John N; Rogers, Craig G; Michalowski, Susan; Zhang, Shaobo; Wang, Mingsheng; Grignon, David J; Cheng, Liang

    2015-11-01

    Clear cell papillary renal cell carcinoma is increasingly recognized as a distinct tumor with unique morphology, immunohistochemistry, and cytogenetics. Histopathology often mimics clear cell renal cell carcinoma; however, metastasis has not been reported, emphasizing the clinical value of recognizing these likely nonaggressive tumors. We studied tumors with borderline morphology of clear cell papillary renal cell carcinoma, utilizing immunohistochemistry and fluorescence in situ hybridization or karyotyping. Tumors from 22 patients (ages 33 to 82 y) were analyzed. Clear cell papillary renal cell carcinoma-like morphology varied from 10% to 90% of the tumor (median 25%). Sources of resemblance included: branched glands (95%), nuclear alignment (68%), small papillary tufts (32%), focal branching papillae (27%), and prominent papillary structures (9%). Carbonic anhydrase IX uniformly revealed diffuse positivity. Staining for cytokeratin 7 (CK7) was focal (64%) or negative (18%) in most tumors (82%); however, >50% labeling was present in 4 (18%). Reactivity for both CD10 and α-methyl-acyl-CoA-racemase (AMACR) was usually present (median 80% and 60% of cells). Seven tumors showed reactivity for high-molecular weight keratin (32%). Chromosome 3p loss was confirmed in 15 tumors (68%), including 4/7 with labeling for high-molecular weight keratin or >50% reactivity for CK7. A discordant immunohistochemical pattern typically correlates with loss of material from chromosome 3p in tumors with incomplete morphology of clear cell papillary renal cell carcinoma, supporting classification as clear cell renal cell carcinoma. Diffuse labeling for CK7 can uncommonly be observed in clear cell renal cell carcinomas confirmed to have chromosome 3p loss, although these do not exhibit the expected staining pattern of clear cell papillary renal cell carcinoma, including positivity for CD10 and AMACR. PMID:26457355

  11. Immunohistochemical (IHC) Analysis of Non-Hodgkin’s Lymphoma (NHL) Spectrum According to WHO/REAL Classification: A Single Centre Experience from Punjab, India

    PubMed Central

    Sharma, Manisha; Mannan, Rahul; Madhukar, Mohit; Navani, Sanjay; Manjari, Mridu; Bhasin, Tejinder Singh; Gill, Karamjit Singh

    2014-01-01

    Introduction: The distribution of the major subtypes of non-Hodgkin’s lymphoma (NHL) differs across geographic regions. This study, from the north Indian state of Punjab, has incorporated immunophenotypic findings while investigating the distribution of NHL subtypes based on World Health Organization (WHO)/ Revised European-American Classification of Lymphoid Neoplasms (REAL) system of classification. Patients and methods: Over all seventy seven cases of lymphoma over a period of one year (between April 2012 and April 2013) were diagnosed in the Department of Pathology, Sri Guru Ram Das Institute of Medical Sciences and Research, Amritsar (Punjab). Of these 30 cases (39%) were of Hodgkin’s Lymphoma (HL) and 47 cases (61%) were of Non Hodgkins lymphoma NHL. Of the total of cases of lympho-proliferative disorders, the diagnosis of NHL was done by light microscopy alone. All the cases diagnosed provisionally as NHL were taken up for immunophenotyping with Immunohistochemical (IHC) studies. There was 100 % concordance between the light microscopy and IHC studies. The individual NHL cases were classified according to the WHO/REAL classification according to the positive or relevant negative immonophenotypic expression and tabulated to ascertain the morphological spectrum of NHL in this part of the country. Results: B-cell lymphomas formed 89.3%, whereas T-cell lymphomas formed 10.7% of the NHLs. Diffuse Large B-Cell Lymphoma (DLBCL) was the most common subtype (46.8% of all NHLs). B-cell small lymphocytic lymphoma, Mantle-Cell Lymphoma (MCL), marginal zone B-cell lymphomas (including MALT lymphomas), Diffuse, mixed small cleaved cell and large-cell type and Follicular centre-cell lymphomas amounted to 17%, 12.8%, 2.1%, 2.1% and 4.3%, respectively. Among the T-cell lymphomas, T-cell lymphoblastic lymphoma, anaplastic large-cell lymphomas of T/null-cell type, and Angioimmunoblastic T-cell lymphoma (AITL) accounted for 6.4%, 2.1%, and 2.1% of all NHL cases

  12. Immunohistochemical analysis of transporters related to clearance of amyloid-β peptides through blood-cerebrospinal fluid barrier in human brain.

    PubMed

    Matsumoto, Koichi; Chiba, Yoichi; Fujihara, Ryuji; Kubo, Hiroyuki; Sakamoto, Haruhiko; Ueno, Masaki

    2015-12-01

    A large number of previous reports have focused on the transport of amyloid-β peptides through cerebral endothelial cells via the blood-brain barrier, while fewer reports have mentioned the transport through the choroid plexus epithelium via the blood-cerebrospinal fluid barrier. Concrete roles of these two pathways remain to be clarified. In this study, we immunohistochemically examined the expression of transporters/receptors that are supposed to be related to the clearance of amyloid-β peptides in the choroid plexus epithelium, the ventricular ependymal cells and the brain microvessels, using seven autopsied human brains. In the choroid plexus epithelium, immunoreactivity for low-density lipoprotein receptor (LDLR), LDLR-related protein 1 (LRP1), LRP2, formylpeptide receptor-like 1 (FPRL1), ATP-binding cassette (ABC) transporter-A1 (ABCA1), ABCC1 and ABCG4 was seen in 7 of 7 brains, while that for ABCB1, ABCG2, RAGE and CD36 was seen in 0-2 brains. In the ventricular ependymal cells, immunoreactivity for CD36, LDLR, LRP1, LRP2, FPRL1, ABCA1, ABCC1 and ABCG4 was seen in 6-7 brains, while that for ABCB1, ABCG2 and RAGE was seen in 0-1 brain. Immunoreactivity for insulin-degrading enzyme (IDE) was seen in three and four brains in the choroid plexus epithelium and the ventricular ependymal cells, respectively. In addition, immunoreactivity for LDLR, ABCB1 and ABCG2 was seen in over 40 % of the microvessels (all seven brains), and that for FPRL1, ABCA1, ABCC1 and RAGE was seen in over 5 % of the microvessels (4-6 brains), while that for CD36, IDE, LRP1, LRP2 and ABCG4 was seen in less than 5 % of the microvessels (0-2 brains). These findings may suggest that these multiple transporters/receptors and IDE expressed on the choroid plexus epithelium, ventricular ependymal cells and brain microvessels complementarily or cooperatively contribute to the clearance of amyloid-β peptides from the brain.

  13. Immunohistochemical Studies of the Kynurenine Pathway in Morphea

    PubMed Central

    Noakes, Rowland; Mellick, Nick

    2013-01-01

    Cutaneous sclerosis, resembling that seen in subcutaneous morphea, is a feature of eosinophilic fasciitis and eosinophilia-myalgia syndrome, two conditions in which the kynurenine pathway is known to be activated. To investigate the possibility of activation of the kynurenine pathway in morphea, skin biopsies were taken from involved and non-involved sites in a series of three patients with morphea. Immunohistochemical stains for quinolinic acid and indoleamine 2,3-dioxygenase (IDO) were performed. PMID:24385714

  14. Immunohistochemical localization and biological activity of the steroidogenic enzyme cytochrome P450 17alpha-hydroxylase/C17, 20-lyase (P450C17) in the frog brain and pituitary.

    PubMed

    Do Rego, Jean Luc; Tremblay, Yves; Luu-The, Van; Repetto, Emanuela; Castel, Hélène; Vallarino, Mauro; Bélanger, Alain; Pelletier, Georges; Vaudry, Hubert

    2007-01-01

    It is now clearly established that the brain has the capability of synthesizing various biologically active steroids including 17-hydroxypregnenolone (17OH-Delta(5)P), 17-hydroxyprogesterone (17OH-P), dehydroepiandrosterone (DHEA) and androstenedione (Delta(4)). However, the presence, distribution and activity of cytochrome P450 17alpha-hydroxylase/C17, 20-lyase (P450(C17)), a key enzyme required for the conversion of pregnenolone (Delta(5)P) and progesterone (P) into these steroids, are poorly documented. Here, we show that P450(C17)-like immunoreactivity is widely distributed in the frog brain and pituitary. Prominent populations of P450(C17)-containing cells were observed in a number nuclei of the telencephalon, diencephalon, mesencephalon and metencephalon, as well as in the pars distalis and pars intermedia of the pituitary. In the brain, P450(C17)-like immunoreactivity was almost exclusively located in neurons. In several hypothalamic nuclei, P450(C17)-positive cell bodies also contained 3beta-hydroxysteroid dehydrogenase-like immunoreactivity. Incubation of telencephalon, diencephalon, mesencephalon, metencephalon or pituitary explants with [(3)H]Delta(5)P resulted in the formation of several tritiated steroids including 17OH-Delta(5)P, 17OH-P, DHEA and Delta(4). De novo synthesis of C(21) 17-hydroxysteroids and C(19) ketosteroids was reduced in a concentration-dependent manner by ketoconazole, a P450(C17) inhibitor. This is the first detailed immunohistochemical mapping of P450(C17) in the brain and pituitary of any vertebrate. Altogether, the present data provide evidence that CNS neurons and pituitary cells can synthesize androgens.

  15. Activation analysis using Cornell TRIGA

    SciTech Connect

    Hossain, Tim Z.

    1994-07-01

    A major use of the Cornell TRIGA is for activation analysis. Over the years many varieties of samples have been analyzed from a number of fields of interest ranging from geology, archaeology and textiles. More recently the analysis has been extended to high technology materials for applications in optical and semiconductor devices. Trace analysis in high purity materials like Si wafers has been the focus in many instances, while in others analysis of major/minor components were the goals. These analysis has been done using the delayed mode. Results from recent measurements in semiconductors and other materials will be presented. In addition the near future capability of using prompt gamma activation analysis using the Cornell cold neutron beam will be discussed. (author)

  16. Standardizing Activation Analysis: New Software for Photon Activation Analysis

    SciTech Connect

    Sun, Z. J.; Wells, D.; Green, J.; Segebade, C.

    2011-06-01

    Photon Activation Analysis (PAA) of environmental, archaeological and industrial samples requires extensive data analysis that is susceptible to error. For the purpose of saving time, manpower and minimizing error, a computer program was designed, built and implemented using SQL, Access 2007 and asp.net technology to automate this process. Based on the peak information of the spectrum and assisted by its PAA library, the program automatically identifies elements in the samples and calculates their concentrations and respective uncertainties. The software also could be operated in browser/server mode, which gives the possibility to use it anywhere the internet is accessible. By switching the nuclide library and the related formula behind, the new software can be easily expanded to neutron activation analysis (NAA), charged particle activation analysis (CPAA) or proton-induced X-ray emission (PIXE). Implementation of this would standardize the analysis of nuclear activation data. Results from this software were compared to standard PAA analysis with excellent agreement. With minimum input from the user, the software has proven to be fast, user-friendly and reliable.

  17. Angiotensin II: Immunohistochemical Study in Sardinian Pterygium

    PubMed Central

    Demurtas, P.; Corrias, M.; Zucca, I.; Piras, F.; Sirigu, P.; Perra, M.T.

    2014-01-01

    The Angiotensin II (Ang II) is the principal effector peptide of the RAS system. It has a pleiotropic effect and, beside its physiological role, it has the property to stimulate angiogenesis and activate multiple signalling pathways related to cell proliferation. The purpose of the study was to determinate the Ang II expression and localization in Sardinian pterygium and normal conjunctiva by immunohistochemistry, and its possible involvement in the development and progression of the disease. Twenty-three pterygiums and eleven normal conjunctiva specimens obtained from Sardinian patients, were processed for paraffin embedding and assessed for the immunohistochemi-cal revelation of Ang II. Significant Ang II expression was identified in pterygium and conjunctiva. Particularly, thirteen pterygium specimens (n=13) displayed exclusively moderate to strong nuclear staining; some specimens (n=5) showed exclusively a moderate cytoplasmic immunoreactivity, and few specimens (n=2) displayed moderate to strong immunoreactivity in both cytoplasm and nucleus. Only 3 specimens were negative. Statistical significance difference in respect of nuclear and cytoplasmic localization was observed between normal conjunctiva and pterygium (P=0.020). The results showed a predominant intranuclear localization of Ang II in pterygium epithelial cells, in spite of conjunctiva that mainly showed cytoplasmic localization. These findings suggest a possible role for Ang II in the development and/or progression of pterygium mediated by the activation of local RAS system. PMID:25308851

  18. Immunohistochemical analysis of bFGF, TGF-beta1 and catalase in rectus abdominis muscle from cattle foetuses at 180 and 260 days post-conception.

    PubMed

    Orzechowski, A; Gajkowska, B; Wojewódzka, U; Cassar-Malek, I; Picard, B; Hocquette, J F

    2002-12-01

    The potential for muscle growth depends on myoblast proliferation, which occurs essentially during the first two thirds of the foetal period in cattle. Thereafter, myofibres acquire their contractile and metabolic properties. Proliferation is regulated by molecular growth factors and by the tissue oxidative activity. The aim of this study was the quantification by immunochemistry of basic fibroblast growth factor (bFGF) and transforming growth factor beta 1 (TGF-beta1) and also of enzyme catalase (CAT) activity in rectus abdominis muscle. Samples were collected from cattle foetuses of different growth potential at 180 and 260 days post-conception (dpc). One major conclusion from this work is that protein contents of the muscle tissue bFGF and, to a lower extent, CAT activity decreased with increasing age during the foetal life. No differences were found between the different genotypes of cattle. However, the CAT to bFGF ratio tended to be lower in fast-growing cattle and increased with foetal age. TGF-beta1 did not change with age and was localised mostly at the vascular bed. CAT was detected in smooth and rough reticulum in striated muscles at 180dpc, and additionally in mitochondria at 260dpc. In conclusion, the balance between intracellular growth factors (bFGF and TGF-beta1) and the activity of antioxidant enzyme CAT may participate in the regulation of the transition from myoblast proliferation to differentiation. Thus, increased ratio of CAT to bFGF might be a good index indicating initiation of muscle maturation in cattle foetus prior to birth.

  19. Effective synaptome analysis of itch-mediating neurons in the spinal cord: A novel immunohistochemical methodology using high-voltage electron microscopy.

    PubMed

    Satoh, Keita; Takanami, Keiko; Murata, Kazuyoshi; Kawata, Mitsuhiro; Sakamoto, Tatsuya; Sakamoto, Hirotaka

    2015-07-10

    Transmission electron microscopy (TEM) is used for three-dimensional (3-D) analysis of synaptic connections in neuroscience research. However, 3-D reconstruction of the synapses using serial ultrathin sections is a powerful but tedious approach requiring advanced technical skills. High-voltage electron microscopy (HVEM) allows examination of thicker sections of biological specimens due to the increased penetration of the more accelerated electrons, which is useful to analyze the 3-D structure of biological specimens. However, it is still difficult to visualize the neural networks and synaptic connections in 3-D using HVEM because of insufficient and non uniform heavy metal staining in the membranous structures in semi-thin sections. Here, we present the successful chemical 3-D neuroanatomy of the rat spinal dorsal horn at the ultrastructural level as a first step for effective synaptome analysis by applying a high-contrast en bloc staining method to immune-HVEM tomography. Our new approach made it possible to examine many itch-mediating synaptic connections and neural networks in the spinal cord simultaneously using HVEM tomography. This novel 3-D electron microscopy is very useful for the analysis of synaptic structure and the chemical neuroanatomy at the 3-D ultrastructural level.

  20. Cartilage Canals in Newborn Dogs: Histochemical and Immunohistochemical Findings

    PubMed Central

    Di Giancamillo, A.; Andreis, M.E.; Taini, P.; Veronesi, M.C.; Di Giancamillo, M.; Modina, S.C.

    2016-01-01

    Cartilage canals (CCs) are microscopic structures involved in secondary ossification centers (SOCs) development. The features of CCs were investigated in the humeral and femoral proximal epiphyses of small-sized newborn dogs (from premature to 28 days after birth) with histochemical and immunohistochemical approaches. Masson’s Trichrome revealed a ring-shaped area around CCs, which changes in colour from green (immature collagen) to red (mature collagen) as ossification progresses; perichondrium staining always matched the ring color. Safranin-O was always negative. Immunohistochemical analysis revealed immunopositivity for both collagen type I and V around the CCs; collagen type II was negative. CCs count showed a tendency to be higher in the humerus than in the femur. This work enlightened for the first time changes in composition of CCs surrounding matrix during SOCs development in dogs, paving the way to further investigations. PMID:27734993

  1. Immunohistochemical prognostic index for breast cancer in young women

    PubMed Central

    Guerra, I; Algorta, J; Díaz de Otazu, R; Pelayo, A; Fariña, J

    2003-01-01

    Aims: Women under 35 years of age comprise a small proportion of patients with breast cancer, but determining their prognosis can be difficult. This prospective, multivariate study looked at several factors with the aim of obtaining a useful index to evaluate the prognosis of these women. Methods: In total, 108 patients below 35 years of age affected by invasive ductal carcinoma without distant metastasis were studied. The mean duration of the follow up period was six years. Histopathological (tumour size, histological grade, and lymph node stage) and immunohistochemical (c-erbB-2, p53, oestrogen receptor, and progesterone receptor) factors were measured in all patients, and the Nottingham prognostic index (NPI) was then calculated. An immunohistochemical prognostic index (IHPI) was created using the arithmetic sum of the four individual immunohistochemical factors. Results: In univariate assessment of survival, all the studied factors yielded a significant association with either overall survival or disease free survival, except for c-erbB-2 and p53 with disease free survival. In univariate calculation of risk, all the factors gave significant results; however, in multivariate analysis only tumour size, histological grade, and progesterone receptor were significant. Both NPI and IHPI correlated significantly with prognosis. In multivariate regression analysis, IHPI correlated with tumour size and there was a significant interaction between both variables. Conclusion: IHPI is very useful in determining the prognosis of tumours ⩽ 2 cm and of moderate use for tumours > 2, although it has no use in tumours > 5 cm. PMID:14645694

  2. Negative pressure wound therapy using gauze and foam: histological, immunohistochemical and ultrasonography morphological analysis of the granulation tissue and scar tissue. Preliminary report of a clinical study.

    PubMed

    Fraccalvieri, Marco; Zingarelli, Enrico; Ruka, Erind; Antoniotti, Umberto; Coda, Renato; Sarno, Antonino; Bocchiotti, Maria Alessandra; Bruschi, Stefano

    2011-08-01

    Negative pressure wound therapy (NPWT) is becoming routine for the preparation of wounds prior to grafting for wound closure. We have been using both foam- and gauze-based NPWT to prepare wounds for closure prior to skin grafting and have obtained similar proportions of closed wounds; 7/7 for wounds treated with gauze-based NPWT and 11/11 for wounds treated with foam-based NPWT. In our follow-up consultations we observed that skin grafts on the foam-treated patients were less pliable than those on the gauze-treated patients. To assess what the mechanism of this effect might be, we compared the specific details of the treatments of both 11 foam and 7 gauze patients, including depth, location, patients' age and co-morbidity; biopsies of granulation and scar tissue were taken and stained with haematoxylin-eosin and by Masson's trichrome staining and conducted ultrasound analysis of the closed wounds, to see if there were features which explained those effects. All foam patients were treated at -125 mm Hg for an average of 25·9 days before skin grafts were applied. All gauze patients were treated at -80 mm Hg for an average of 24·7 days before skin grafts were applied. Biopsies of granulation tissue prior to skin grafting from five foam and four gauze-based NPWT patients did not reveal any obvious histological differences between the treatments. Ultrasound analysis of the skin-grafted wounds showed an average depth of scar tissue of 18 mm in the wound beds of the foam-treated wounds and 7 mm in the gauze-treated ones. Biopsies taken on the scar tissue after treatment with the gauze showed a minor tissue thickness and disorganisation and less sclerotic components. The findings of this preliminary analysis suggest that foam-based NPWT may induce a thicker layer of scar tissue beneath skin grafts than gauze-based NPWT which might explain a reduced pliability of the reconstructed bed. At present it is unclear which mechanism might be responsible for the difference in

  3. Utility of immunohistochemical investigation of SDHB and molecular genetic analysis of SDH genes in the differential diagnosis of mesenchymal tumors of GIT.

    PubMed

    Dubova, M; Sedivcova, M; Michal, M; Kokoskova, B; Ryska, A; Smid, D; Daum, O

    2015-02-01

    Loss of expression of beta subunit of succinate dehydrogenase (SDHB) was proved to be present in a subgroup of KIT/PDGFRA wt gastrointestinal stromal tumors (GISTs). To evaluate possible diagnostic utility of SDHB immunohistochemistry in the differential diagnostics of mesenchymal tumors of gastrointestinal tract (GIT), 11 cases of KIT/PDGFRA wt GISTs, 12 gastric schwannomas (GSs), 20 solitary fibrous tumors (SFTs), 4 leiomyomas (LMs), 16 leiomyosarcomas (LMSs), 5 synovial sarcomas (SSs), 3 endometrioid stromal sarcomas (ESSs), and 1 ileal inflammatory myofibroblastic tumor (IMT) were investigated for SDHB immunoexpression together with molecular genetic analysis of genes encoding succinate dehydrogenase (SDH). Three recent cases of KIT/PDGFRA mutant GISTs were used as controls. Among the 11 KIT/PDGFRA wt GISTs, 6 expressed SDHB, 1 of them harboring a sequence change of SDHD. All SDHB-negative cases were SDHB-D wt. In 1 of the control GIST cases molecular genetic analysis revealed an SDHD sequence change in addition to a mutation in KIT exon 11. No SFT was truly SDHB-negative, but in 2 of them the staining was impossible to analyze. Furthermore, 1 SFT carried an SDHB and another 1 SDHD sequence change. All GSs, LMs, LMSs, SSs, ESSs, and IMT were SDHB-positive or non-analyzable, and SDHB-D wt. Additional factors may play a role in regulating expression of SDHB. Furthermore, SDHB immunohistochemistry alone may be misleading in excluding tumors other than GIST (especially SFT) in the differential diagnosis of KIT/PDGFRA wt mesenchymal tumors of GIT.

  4. Cannabinoid CB2 Receptors in a Mouse Model of Aβ Amyloidosis: Immunohistochemical Analysis and Suitability as a PET Biomarker of Neuroinflammation

    PubMed Central

    Wang, Yuchuan; Ravert, Hayden; Gao, Yongjun; Koppel, Jeremy; Lee, Deidre; Pletnikova, Olga; Cho, Eugenia; Sayyida, Nuzhat; Hiatt, Andrew; Troncoso, Juan; Davies, Peter; Dannals, Robert F.; Pomper, Martin G.; Horti, Andrew G.

    2015-01-01

    In Alzheimer’s disease (AD), one of the early responses to Aβ amyloidosis is recruitment of microglia to areas of new plaque. Microglial receptors such as cannabinoid receptor 2 (CB2) might be a suitable target for development of PET radiotracers that could serve as imaging biomarkers of Aβ-induced neuroinflammation. Mouse models of amyloidosis (J20APPswe/ind and APPswe/PS1ΔE9) were used to investigate the cellular distribution of CB2 receptors. Specificity of CB2 antibody (H60) was confirmed using J20APPswe/ind mice lacking CB2 receptors. APPswe/PS1ΔE9 mice were used in small animal PET with a CB2-targeting radiotracer, [11C]A836339. These studies revealed increased binding of [11C]A836339 in amyloid-bearing mice. Specificity of the PET signal was confirmed in a blockade study with a specific CB2 antagonist, AM630. Confocal microscopy revealed that CB2-receptor immunoreactivity was associated with astroglial (GFAP) and, predominantly, microglial (CD68) markers. CB2 receptors were observed, in particular, in microglial processes forming engulfment synapses with Aβ plaques. In contrast to glial cells, neuron (NeuN)-derived CB2 signal was equal between amyloid-bearing and control mice. The pattern of neuronal CB2 staining in amyloid-bearing mice was similar to that in human cases of AD. The data collected in this study indicate that Aβ amyloidosis without concomitant tau pathology is sufficient to activate CB2 receptors that are suitable as an imaging biomarker of neuroinflammation. The main source of enhanced CB2 PET binding in amyloid-bearing mice is increased CB2 immunoreactivity in activated microglia. The presence of CB2 immunoreactivity in neurons does not likely contribute to the enhanced CB2 PET signal in amyloid-bearing mice due to a lack of significant neuronal loss in this model. However, significant loss of neurons as seen at late stages of AD might decrease the CB2 PET signal due to loss of neuronally-derived CB2. Thus this study in mouse models

  5. Immunohistochemical analysis of MMP-9, MMP-2 and TIMP-1, TIMP-2 expression in the central nervous system following infection with viral and bacterial meningitis.

    PubMed

    Sulik, Artur; Chyczewski, Lech

    2008-01-01

    Matrix metalloproteinases (MMPs) are capable of degrading components of the basal lamina of cerebral vessels, thereby disrupting the blood-brain barrier and inducing leukocyte recruitment. This study provides comprehensive information regarding the cell specificity of matrix metalloproteinases (MMP-2, MMP-9) and their binding tissue inhibitors (TIMP-1, TIMP-2) in the central nervous system during viral and bacterial meningitis. Specifically, we evaluated the immunoreactivity of MMPs and TIMPs in various cell types in brain parenchyma and meninges obtained from autopsy tissues. We found that a higher proportion of endothelial cells were positive for MMP-9 during meningitis when compared to controls. In addition, the immunoreactivity of MMP-9 decreased and the immunoreactivity of TIMP-1 increased in astrocytes upon infection. Furthermore, the results of this study revealed that mononuclear cells were highly immunoreactive for TIMP-1, TIMP-2 and MMP-9 during viral meningitis and that the expression of TIMPs in polymorphonuclear cells was even higher during bacterial meningitis. Taken together the results of this study indicated that the central nervous system resident cells and inflammatory infiltrates contribute to MMPs activity and that the expression patterns vary between cell types and in response to viral and bacterial meningitis.

  6. The Expression Patterns of ER, PR, HER2, CK5/6, EGFR, Ki-67 and AR by Immunohistochemical Analysis in Breast Cancer Cell Lines

    PubMed Central

    Subik, Kristina; Lee, Jin-Feng; Baxter, Laurie; Strzepek, Tamera; Costello, Dawn; Crowley, Patti; Xing, Lianping; Hung, Mien-Chie; Bonfiglio, Thomas; Hicks, David G.; Tang, Ping

    2010-01-01

    The molecular classification for breast carcinomas has been used in clinical studies with a simple surrogate panel of immunohistochemistry (IHC) markers. The objective of this current project was to study the molecular classification of commonly used breast cancer cell lines by IHC analysis. Seventeen breast cancer cell lines were harvested, fixed in formalin and made into cell blocks. IHC analyses were performed on each cell block with antibodies to estrogen receptor (ER), progesterone receptor (PR), HER2, EGFR, CK5/6, Ki-67 and androgen receptor (AR). Among the 17 cell lines, MCF-7 and ZR-75-1 fell to Luminal A subtype; BT-474 to Luminal B subtype; SKBR-3, MDA-MD-435 and AU 565 to HER2 over-expression subtype; MDA-MB-231, MCF-12A, HBL 101, HS 598 T, MCF-10A, MCF-10F, BT-20, 468 and BT-483 to basal subtype. MDA-MB-453 belonged to Unclassified subtype. Since each subtype defined by this IHC-based molecular classification does show a distinct clinical outcome, attention should be paid when choosing a cell line for any study. PMID:20697531

  7. Atypical lipomatous tumor/"well-differentiated liposarcoma" of the skin clinically presenting as a skin tag: clinicopathologic, immunohistochemical, and molecular analysis of 2 cases.

    PubMed

    Paredes, Bruno E; Mentzel, Thomas

    2011-08-01

    Liposarcomas are extremely rare in the skin. When they involve the skin, it is usually by upward spread from a subcutaneous or deeper seated liposarcoma. Very rarely, liposarcoma metastasize to the skin or arise as a primary dermal lesion. We describe 2 cases of atypical lipomatous tumor "well-differentiated liposarcoma" located in dermis. Both presented clinically as a skin tag. The neoplasms arose in a 56-year-old female and a 69-year-old male patient. Both lesions were treated by excision and reexcision. In addition to classical morphology of atypical lipomatous tumor with evidence of lipoblasts and atypical adipocytes, immunohistochemistry with nuclear murine double-minute type 2 protein and cyclin-dependent kinase-4 expression as well as fluorescence in situ hybridization analysis showing an amplification of murine double-minute type 2 protein and cyclin-dependent kinase-4 were helpful to establish the diagnosis. None of the cases recurred after surgical treatment. These 2 cases show the importance of not to misdiagnose lesions which clinically may appear to be benign.

  8. Atypical lipomatous tumor/"well-differentiated liposarcoma" of the skin clinically presenting as a skin tag: clinicopathologic, immunohistochemical, and molecular analysis of 2 cases.

    PubMed

    Paredes, Bruno E; Mentzel, Thomas

    2011-08-01

    Liposarcomas are extremely rare in the skin. When they involve the skin, it is usually by upward spread from a subcutaneous or deeper seated liposarcoma. Very rarely, liposarcoma metastasize to the skin or arise as a primary dermal lesion. We describe 2 cases of atypical lipomatous tumor "well-differentiated liposarcoma" located in dermis. Both presented clinically as a skin tag. The neoplasms arose in a 56-year-old female and a 69-year-old male patient. Both lesions were treated by excision and reexcision. In addition to classical morphology of atypical lipomatous tumor with evidence of lipoblasts and atypical adipocytes, immunohistochemistry with nuclear murine double-minute type 2 protein and cyclin-dependent kinase-4 expression as well as fluorescence in situ hybridization analysis showing an amplification of murine double-minute type 2 protein and cyclin-dependent kinase-4 were helpful to establish the diagnosis. None of the cases recurred after surgical treatment. These 2 cases show the importance of not to misdiagnose lesions which clinically may appear to be benign. PMID:21358383

  9. Neutron Activation Analysis: Techniques and Applications

    NASA Astrophysics Data System (ADS)

    MacLellan, Ryan

    2011-04-01

    The role of neutron activation analysis in low-energy low-background experimentsis discussed in terms of comparible methods. Radiochemical neutron activation analysis is introduce. The procedure of instrumental neutron activation analysis is detailed especially with respect to the measurement of trace amounts of natural radioactivity. The determination of reactor neutron spectrum parameters required for neutron activation analysis is also presented.

  10. Neutron Activation Analysis: Techniques and Applications

    SciTech Connect

    MacLellan, Ryan

    2011-04-27

    The role of neutron activation analysis in low-energy low-background experimentsis discussed in terms of comparible methods. Radiochemical neutron activation analysis is introduce. The procedure of instrumental neutron activation analysis is detailed especially with respect to the measurement of trace amounts of natural radioactivity. The determination of reactor neutron spectrum parameters required for neutron activation analysis is also presented.

  11. Immunohistochemical Characterization of Large Intestinal Adenocarcinoma in the Rhesus Macaque (Macaca mulatta).

    PubMed

    Harbison, C E; Taheri, F; Knight, H; Miller, A D

    2015-07-01

    In rhesus macaques, adenocarcinomas of either the ileocecal junction or colon are common spontaneous tumors in aging populations. The macaque tumors have similar gross and histologic characteristics compared with their human counterpart, but little is known regarding the immunohistochemical expression of proteins that are commonly implicated in the pathogenesis of these tumors in humans. We performed a retrospective review of 22 cases of large intestinal carcinoma in the rhesus macaque and evaluated the expression pattern of a panel of potentially prognostically significant proteins identified from human studies. Histologic characteristics of the tumors included abundant mucin deposition, transmural spread, and lymphatic invasion. All rhesus adenocarcinomas displayed altered expression of 1 or more of CD10, β-catenin, sirtuin 1, cytokeratin 17, and p53 compared with age-matched controls. Zymographic analysis of active matrix metalloproteinases 2 and 9 in the serum from 5 animals failed to reveal statistically significant differences between adenocarcinoma cases and controls. Based on the data presented herein, large intestinal carcinomas in the macaque share many histomorphologic and immunohistochemical similarities to large intestinal tumors in humans. Further validation of this animal model is considered important for the development of novel therapeutics and a better understanding of the pathogenesis.

  12. US--ITER activation analysis

    SciTech Connect

    Attaya, H.; Gohar, Y.; Smith, D.

    1990-09-01

    Activation analysis has been made for the US ITER design. The radioactivity and the decay heat have been calculated, during operation and after shutdown for the two ITER phases, the Physics Phase and the Technology Phase. The Physics Phase operates about 24 full power days (FPDs) at fusion power level of 1100 MW and the Technology Phase has 860 MW fusion power and operates for about 1360 FPDs. The point-wise gamma sources have been calculated everywhere in the reactor at several times after shutdown of the two phases and are then used to calculate the biological dose everywhere in the reactor. Activation calculations have been made also for ITER divertor. The results are presented for different continuous operation times and for only one pulse. The effect of the pulsed operation on the radioactivity is analyzed. 6 refs., 12 figs., 1 tab.

  13. Prompt-gamma activation analysis

    SciTech Connect

    Lindstrom, R.M.

    1993-01-01

    A permenent, full-time instrument for prompt-gamma activation analysis is nearing completion as part of the Cold Neutron Research Facility (CNRF). The design of the analytical system has been optimized for high gamma detection efficiency and low background, particularly for hydrogen. Because of the purity of the neutron beam, shielding requirements are modest and the scatter-capture background is low. As a result of a compact sample-detector geometry, the sensitivity (counting rate per gram of analyte) is a factor of four better than the existing Maryland-NIST thermal-neutron instrument at the reactor. Hydrogen backgrounds of a few micrograms have already been achieved, which promises to be of value in numerous applications where quantitative nondestructive analysis of small quantities of hydrogen in materials is necessary.

  14. Malignant myoepithelioma of the salivary glands: clinicopathological and immunohistochemical features.

    PubMed

    Dean, A; Sierra, R; Alamillos, F J; Lopez-Beltran, A; Morillo, A; Arévalo, R; Rodas, J; Ruiz-Masera, J J; García-Lopez, A

    1999-02-01

    Malignant myoepitheliomas (myoepithelial carcinomas) are uncommon, and we know of only 29 reported cases. We present a new case together with its clinical, histological, and immunohistochemical features. The tumour was located in the inferior vestibular sulcus of a 64-year-old woman. She was treated by wide local resection. Malignant myoepitheliomas are distinguished from benign myoepithelial neoplasms by their infiltrating and destructive growth. The tumour cells may be spindle-shaped or more rounded (plasmacytoid cells) and contain cellular pleomorphism and mitotic activity. The clinical and biological behaviour of this tumour is not yet known and there is little information about treatment and prognosis.

  15. Histochemical and immunohistochemical characteristics of elastofibromas.

    PubMed

    Tasli, F; Vardar, E; Argon, A; Kabat, T; Deniz, S; Nart, A; Kececi, Y

    2014-06-01

    Elastofibromas are slow-growing and rare soft-tissue tumors. The etiology and pathogenetic mechanisms are still controversial and there are only a few studies in the literature investigating the histochemical, immunohistochemical, and genetic features to determine the pathogenesis. We investigated the cellular composition of lesions with a diagnosis of elastofibroma in 17 patients by using histochemical and immunohistochemical methods. There were 17 cases with a mean age of 53.5 years. Mean lesion diameter was 6.6 cm. The immunohistochemical method showed vimentin and factor XIIIa positivity in all cases. Four cases had focal myoglobin positivity in the spindle-shaped cells between the collagen fibers. Spindle cells were positive for CD34 in 8 cases. Smooth muscle actin, desmin, type 4 collagen and laminin were negative in all cases. The elastic nature of the abnormal fibers was shown histoch with Verhoeff elastin staining and aldehyde fuchsin staining in all cases. Our results have shown that the concurrent positivity of factor XIIIa and CD34 in the cells forming the lesion might show that the lesionoriginates from primitive dermal mesenchymal cells. In addition, the myoglobin positivity found in some cases indicates the possibility of a myofibroblastic origin of elastofibromas. PMID:25119171

  16. Assessment of candidate immunohistochemical prognostic markers of meningioma recurrence.

    PubMed

    Csonka, T; Murnyák, B; Szepesi, R; Bencze, J; Bognár, L; Klekner, A; Hortobágyi, T

    2016-01-01

    Although tumour recurrence is an important and not infrequent event in meningiomas, predictive immunohistochemical markers have not been identified yet. The aim of this study was to address this clinically relevant problem by systematic retrospective analysis of surgically completely resected meningiomas with and without recurrence, including tumour samples from patients who underwent repeat surgeries. Three established immunohistochemical markers of routine pathological meningioma work-up have been assessed: the proliferative marker Ki-67 (clone Mib1), the tumour suppressor gene p53 and progesterone receptor (PR). All these proteins correlate with the tumour WHO grade, however the predictive value regarding recurrence and progression in tumour grade is unknown. One hundred and fourteen surgical specimens of 70 meningioma patients (16 male and 54 female) in a 16 years' interval have been studied. All tumours had apparently complete surgical removal. On Mib1, PR and p53 immunostained sections, the percentage of labelled tumour cells, the staining intensity and the multiplied values of these parameters (the histoscore) was calculated. Results were statistically correlated with tumour WHO grade, (sub)type, recurrence and progression in WHO grade at subsequent biopsies. Our results confirmed previous findings that the WHO grade is directly proportional to Mib1 and p53 and is inversely proportional to the PR immunostain. We have demonstrated that Mib1 and p53 have a significant correlation with and predictive value of relapse/recurrence irrespective of the histological subtype of the same WHO grade. As a quantitative marker, Mib1 has the best correlation with a percentage of labelled cells, whereas p53 with intensity and histoscore. In conclusion, the immunohistochemical panel of PR, p53, Mib1 in parallel with applying standard diagnostic criteria based on H and E stained sections is sufficient and reliable to predict meningioma recurrence in surgically completely

  17. Current concepts in the immunohistochemical evaluation of liver tumors.

    PubMed

    Koehne de Gonzalez, Anne K; Salomao, Marcela A; Lagana, Stephen M

    2015-06-01

    Immunohistochemistry often plays an important role in the evaluation of liver tumors. Recent advances have established a classification system for hepatocellular adenomas (HCAs) based on morphology, molecular alterations, and immunohistochemistry. Specifically, loss of liver fatty acid binding protein is seen in HNF1α-inactivated HCA, staining with serum amyloid A is seen in inflammatory HCA, and diffuse staining with glutamine synthetase (GS) is seen in β-catenin activated HCA. A panel of immunohistochemical stains including glypican-3 (GPC-3), heat shock protein 70, and GS are useful in distinguishing HCC from non-malignant dysplastic nodules. Immunohistochemistry is also useful to determine whether a liver tumor is of primary hepatocellular or metastatic origin. Recently described markers useful for this purpose include arginase-1, GPC-3, and bile salt export pump. These newer markers may offer superior utility when compared to traditional markers of hepatocellular differentiation such as alpha-fetoprotein, hepatocyte paraffin-1, polyclonal carcinoembryonic antigen, and CD10. This paper will review recent advances in the immunohistochemical evaluation of liver tumors. PMID:26052385

  18. Histological and immunohistochemical studies on primary intracranial canine histiocytic sarcomas

    PubMed Central

    THONGTHARB, Atigan; UCHIDA, Kazuyuki; CHAMBERS, James Kenn; KAGAWA, Yumiko; NAKAYAMA, Hiroyuki

    2015-01-01

    Histiocytic sarcoma is a progressive and fatal malignant neoplasm that mainly occurs in middle- to old-aged dogs. This study describes clinicopathological, histological and immunohistochemical characteristics of intracranial histiocytic sarcomas in 23 dogs. Magnetic resonance imaging and/or computed tomography of the brains revealed that the tumors mainly located in the cerebrum, particularly the frontal lobe. Seizure was a predominant clinical sign in most of the cases. Histologically, the tumor cells were morphologically classified into round/polygonal- and spindle-shaped cell types. There was a significant association between tumor cell types and hemophagocytic activity (P<0.05). However, there was no significant difference in other clinicopathological parameters and mitotic index between the 2 types. Immunohistochemically, tumor cells were strongly positive for HLA-DR, Iba-1 and CD204 in all the 23 cases, for iNOS in 20, for CD163 in 17, for CD208 (DC-LAMP) in 9, for lysozyme in 8 and for S100 in 5 cases. In addition, the Ki67-proliferative index showed range of 0.50–64.33% (Average 26.60 ± 3.81%). These observations suggest that canine primary intracranial histiocytic sarcomas tend to exhibit both dendritic cell and macrophage phenotypes of histiocytic differentiation. PMID:26668164

  19. Immunohistochemical diagnostic and prognostic markers for melanoma.

    PubMed

    Nosrati, Mehdi; Kashani-Sabet, Mohammed

    2014-01-01

    Recent studies in our laboratory have identified novel molecular diagnostic and prognostic markers based on analyses in large cohorts of melanoma patients. These markers were initially derived from gene expression profiling analyses of distinct stages of melanoma progression. Immunohistochemical analyses confirmed the differential expression of these markers, and immunohistochemistry-based multimarker assays were developed to assess melanoma diagnosis and prognosis at the molecular level. In this chapter we review the development of these assays and the methodologies used to assess marker expression in both nevi and primary melanomas. PMID:24258983

  20. Immunohistochemical and biochemical assay of versican in human sound predentine/dentine matrix

    PubMed Central

    Ruggeri, A.; Orsini, G.; Mazzoni, A.; Nato, F.; Papa, V.; Piccirilli, M.; Putignano, A.; Mazzotti, G.; De Stefano Dorigo, E.; Breschi, L.

    2009-01-01

    Aim of this study was to investigate the distribution of versican proteoglycan within the human dentine organic matrix by means of a correlative immunohistochemical analysis with field emission in-lens scanning electron microscope (FEI-SEM), transmission electron microscope (TEM), fluorescence microscope (FM) and biochemical assay. Specimens containing dentine and predentine were obtained from non carious human teeth and divided in three groups: 1) FEI-SEM group: sections were exposed to a pre-embedding immunohistochemical procedure; 2) TEM group: specimens were fixed, demineralised, embedded and submitted to a post-embedding immunohistochemical procedure; 3) FM group: sections mineralised and submitted to a pre-embedding immunohistochemical procedure with fluorescence labelling. Specimens were exposed to two different antibodies to assay distribution of versican fragments and whole versican molecule. Western Blotting analysis of dentine and pulp extracts was also performed. The correlative FEI-SEM,TEM and FM analysis revealed positive immunoreaction for versican fragments both in predentine and dentine, while few gold particles identifying the whole versican molecule were found in predentine only under TEM. No labelling of versican whole molecule was detected by FEI-SEM and FM analysis. The immunoblotting analysis confirmed the morphological findings. This study suggests that in fully developed human teeth versican fragments are significant constituents of the human dentine and predentine organic matrix, while versican whole molecule can be visualised in scarce amount within predentine only. The role of versican fragments within human dentine organic matrix should be further elucidated.

  1. Successful Treatment of Granuloma Faciale with Topical Tacrolimus: A Case Report and Immunohistochemical Study

    PubMed Central

    Tojo, Gen-ichi; Fujimura, Taku; Kambayashi, Yumi; Kikuchi, Katsuko; Aiba, Setsuya

    2012-01-01

    We report the case of a 55-year-old Japanese patient with granuloma faciale (GF) successfully treated with topical tacrolimus and describe the immunohistochemical study. Immunohistochemical staining revealed that the patient's granuloma contained CD3+, CD4+, CD8+, CD68+ and CD163+ cells. Interestingly, these cells contained granulysin+ T cells and lacked Foxp3high+ regulatory T cells. In addition, the macrophages were mainly CD163+, which suggested that the alternatively activated macrophage is one of the main components of GF. In summary, the present data shed light on the granuloma-composing cells and possible mechanisms in the treatment of GF with topical tacrolimus. PMID:22807901

  2. Immunohistochemical techniques for the human inner ear.

    PubMed

    Lopez, Ivan A; Ishiyama, Gail; Hosokawa, Seiji; Hosokawa, Kumiko; Acuna, Dora; Linthicum, Fred H; Ishiyama, Akira

    2016-10-01

    In this review, we provide a description of the recent methods used for immunohistochemical staining of the human inner ear using formalin-fixed frozen, paraffin and celloidin-embedded sections. We also show the application of these immunohistochemical methods in auditory and vestibular endorgans microdissected from the human temporal bone. We compare the advantages and disadvantages of immunohistochemistry (IHC) in the different types of embedding media. IHC in frozen and paraffin-embedded sections yields a robust immunoreactive signal. Both frozen and paraffin sections would be the best alternative in the case where celloidin-embedding technique is not available. IHC in whole endorgans yields excellent results and can be used when desiring to detect regional variations of protein expression in the sensory epithelia. One advantage of microdissection is that the tissue is processed immediately and IHC can be made within 1 week of temporal bone collection. A second advantage of microdissection is the excellent preservation of both morphology and antigenicity. Using celloidin-embedded inner ear sections, we were able to detect several antigens by IHC and immunofluorescence using antigen retrieval methods. These techniques, previously applied only in animal models, allow for the study of numerous important proteins expressed in the human temporal bone potentially opening up a new field for future human inner ear research. PMID:27480257

  3. Human vomeronasal epithelium development: An immunohistochemical overview.

    PubMed

    Dénes, Lóránd; Pap, Zsuzsanna; Szántó, Annamária; Gergely, István; Pop, Tudor Sorin

    2015-06-01

    The vomeronasal organ (VNO) is the receptor structure of the vomeronasal system (VNS) in vertebrates. It is found bilaterally in the submucosa of the inferior part of the nasal septum. There are ongoing controversies regarding the functionality of this organ in humans. In this study we propose the immunohistochemical evaluation of changes in components of the human vomeronasal epithelium during foetal development. We used 45 foetuses of different age, which were included in three age groups. After VNO identification immunohistochemical reactions were performed using primary antibodies against the following: neuron specific enolase, calretinin, neurofilament, chromogranin, synaptophysin, cytokeratin 7, pan-cytokeratin and S100 protein. Digital slides were obtained and following colorimetric segmentation, surface area measurements were performed. The VNO was found in less than half of the studied specimens (42.2%). Neuron specific enolase and calretinin immunoexpression showed a decreasing trend with foetal age, while the other neural/neuroendocrine markers were negative in all specimens. Cytokeratin 7 expression increased with age, while Pan-Ctk had no significant variations. S100 protein immunoexpression also decreased around the VNO. The results of the present work uphold the theory of regression of the neuroepithelium that is present during initial stages of foetal development.

  4. Histological and immunohistochemical analyses of the chronology of healing process after immediate tooth replantation in incisor rat teeth.

    PubMed

    Panzarini, Sônia Regina; Okamoto, Roberta; Poi, Wilson Roberto; Sonoda, Celso Koogi; Pedrini, Denise; da Silva, Paula Ervolino; Saito, Celia Tomiko Matida Hamata; Marão, Heloísa Fonseca; Sedlacek, Paulo

    2013-02-01

    Dental tissues have special characteristics, and its regenerative capacity is noteworthy. However, understanding the circumstances that lead to regeneration is challenging. In this study, the chronology of the healing process after immediate replantation of rat incisor teeth was examined by histological and immunohistochemical analyses within a 60-day period. Thirty-six male Wistar rats had their maxillary right incisors extracted and replanted after 15 min in saline storage. The rats were sacrificed immediately 3, 7, 15, 28, and 60 days after replantation. The histological analysis showed rupture of the periodontal ligament and formation of a blood clot, which started being replaced by a connective tissue after 3 days. At 7 days, the gingival mucosa epithelium was reinserted and areas of root resorption could be seen. At 15 days, the periodontal ligament was repaired. At 3 days, the pulp presented an absence of the odontoblast layer, which started being replaced by a connective tissue. This tissue suffered gradual calcification, filling the root canal at 28 and 60 days. The root ends were closed. The immunohistochemical analysis revealed greater expression of OP, OPG, and RANK proteins in the initial periods (0 and 3 days), while TRAP expression predominated at 28 and 60 days (P < 0.05). In conclusion, in delayed tooth replantation, there is great new bone formation activity in the earlier periods of the repair process, while a predominance of bone resorption and remodeling is observed in the more advanced periods.

  5. Total body nitrogen analysis. [neutron activation analysis

    NASA Technical Reports Server (NTRS)

    Palmer, H. E.

    1975-01-01

    Studies of two potential in vivo neutron activation methods for determining total and partial body nitrogen in animals and humans are described. A method using the CO-11 in the expired air as a measure of nitrogen content was found to be adequate for small animals such as rats, but inadequate for human measurements due to a slow excretion rate. Studies on the method of measuring the induced N-13 in the body show that with further development, this method should be adequate for measuring muscle mass changes occurring in animals or humans during space flight.

  6. Analysis of active renin heterogeneity.

    PubMed

    Katz, S A; Malvin, R L; Lee, J; Kim, S H; Murray, R D; Opsahl, J A; Abraham, P A

    1991-09-01

    Active renin is a heterogeneous enzyme that can be separated into multiple forms with high-resolution isoelectric focusing. The isoelectric heterogeneity may result from differences in glycosylation between the different forms. In order to determine the relationship between active renin heterogeneity and differences in composition or attachment of oligosaccharides, two separate experiments were performed: (i) Tunicamycin, which interferes with normal glycosylation processing, increased the proportion of relatively basic renin forms secreted into the incubation media by rat renal cortical slices. (ii) Endoglycosidase F, which enzymatically removes carbohydrate from some classes of glycoprotein, similarly increased the proportion of relatively basic forms when incubated with active human recombinant renin. In addition, further studies with inhibitors of human renin activity revealed that the heterogeneous renin forms were similarly inhibited by two separate renin inhibitors. These results are consistent with the hypothesis that renin isoelectric heterogeneity is due in part to differences in carbohydrate moiety attachment and that the heterogeneity of renin does not influence access of direct renin inhibitors to the active site of renin.

  7. Analysis of active renin heterogeneity.

    PubMed

    Katz, S A; Malvin, R L; Lee, J; Kim, S H; Murray, R D; Opsahl, J A; Abraham, P A

    1991-09-01

    Active renin is a heterogeneous enzyme that can be separated into multiple forms with high-resolution isoelectric focusing. The isoelectric heterogeneity may result from differences in glycosylation between the different forms. In order to determine the relationship between active renin heterogeneity and differences in composition or attachment of oligosaccharides, two separate experiments were performed: (i) Tunicamycin, which interferes with normal glycosylation processing, increased the proportion of relatively basic renin forms secreted into the incubation media by rat renal cortical slices. (ii) Endoglycosidase F, which enzymatically removes carbohydrate from some classes of glycoprotein, similarly increased the proportion of relatively basic forms when incubated with active human recombinant renin. In addition, further studies with inhibitors of human renin activity revealed that the heterogeneous renin forms were similarly inhibited by two separate renin inhibitors. These results are consistent with the hypothesis that renin isoelectric heterogeneity is due in part to differences in carbohydrate moiety attachment and that the heterogeneity of renin does not influence access of direct renin inhibitors to the active site of renin. PMID:1908097

  8. Neutron activation analysis of a penny

    NASA Astrophysics Data System (ADS)

    Stevens, Richard E.

    2000-04-01

    Neutron activation analysis has been used for many years as an analysis tool and as an educational tool to teach students about nuclear properties. This article presents an exercise in the neutron activation analysis of a penny which, due to the simplicity of the resulting gamma-ray spectra, is appropriate for general physics classes. Students express a great deal of interest both in seeing the reactor in use as well as determining the composition of something that is familiar to them.

  9. Primary amyloidosis A. Immunohistochemical and biochemical characterization.

    PubMed Central

    Picken, M. M.; Pelton, K.; Frangione, B.; Gallo, G.

    1987-01-01

    Primary "idiopathic" amyloidosis is usually related to immunoglobulin light chain (AL) associated with immunocytic dyscrasias, while secondary "reactive" amyloidosis (AA) is related to serum amyloid A protein (SAA) and typically occurs with chronic inflammation, malignancy, or familial Mediterranean fever. In the present study, amyloid fibril protein extracted from frozen and paraffin-embedded tissue from a patient (CAR) with primary systemic amyloidosis proved to be AA protein by immunohistochemical, immunochemical, and amino terminal sequence. Extracts from both frozen and formalin-fixed paraffin-embedded kidney and spleen yielded similar monomers and dimers of the AA protein. The additional high-molecular-weight bands and a distinct 12,000-dalton fragment in the amyloid protein extracted from the formalin-fixed paraffin-embedded lung suggest that different processing of proteins, ie, by polymerization and/or degradation, may occur in different organs. Images Figure 1 Figure 2 Figure 3 PMID:3425691

  10. LACBWR primary shield activation analysis

    SciTech Connect

    Nelson, L.L.; Lahti, G.P.; Johnson, W.J.

    1996-11-01

    Nuclear power plants in the US are required to estimate the costs of decommissioning to ensure that adequate funds are accumulated during the useful life of the plant. A major component of the decommissioning cost is the disposal of radioactive material, including material near the reactor created by neutron activation. An accurate assessment of the residual radioactivity in the reactor`s primary shield is necessary to determine this portion of the decommissioning demolition and disposal cost. This paper describes the efforts used to determine the activation levels remaining in the primary shield of the LaCrosse boiling water reactor (LACBWR), owned and operated by Dairyland Power Cooperative.

  11. Conference on Instrumental Activation Analysis: IAA 89

    NASA Astrophysics Data System (ADS)

    Vobecky, M.; Obrusnik, I.

    1989-05-01

    The proceedings contain 40 abstracts of papers all of which have been incorporated in INIS. The papers were centred on the applications of radioanalytical methods, especially on neutron activation analysis, x ray fluorescence analysis, PIXE analysis and tracer techniques in biology, medicine and metallurgy, measuring instruments including microcomputers, and data processing methods.

  12. Adrenal lymphangioma: clinicopathologic and immunohistochemical characteristics of a rare lesion.

    PubMed

    Ellis, Carla L; Banerjee, Priya; Carney, Erin; Sharma, Rajni; Netto, George J

    2011-07-01

    Adrenal lymphangiomas, also known as cystic adrenal lymphangiomas, are rare, benign vascular lesions that usually remain asymptomatic throughout life. Although previously adrenal lymphangioma lesions were primarily found at autopsy, they are currently detected during imaging work-up for unrelated causes and are likely to imitate other adrenocortical or adrenal medullary neoplasms. We aimed to retrospectively review all adrenal lymphangioma cases at our hospital and further document their lymphatic origin by immunohistochemical staining. A search of surgical pathology records (1984-2008) was conducted. All hematoxylin and eosin sections were retrieved from archives and reviewed by 2 pathologists in the study. Clinical information was gathered from electronic medical records. Representative paraffin-embedded sections from each case were selected for immunohistochemical analysis using monoclonal antibodies D2-40 and AE1/AE3. A total of 9 adrenal lymphangioma cases were identified (6 women and 3 men). All 9 patients were adults at time of diagnosis with a mean age of 42 years (range, 28-56 years). There were 7 white patients, 1 African American patient, and 1 Asian patient. The average size of an adrenal lymphangioma lesion was 4.9 cm (range, 2.0-13.5 cm). Adrenal lymphangioma was twice more frequently located on the right side (6 right-sided and 3 left-sided). Clinically, 4 (44%) of the 9 lesions presented with abdominal, flank, or back pain. One lymphangioma was found during work-up for labile hypertension. The remaining 4 lesions (44%) were asymptomatic and incidentally found during imaging studies for unrelated causes. Surgical removal was achieved by total adrenalectomy in 8 of the 9 lesions and by partial adrenalectomy in the remaining case. No evidence of recurrence or development of a contralateral lesion was encountered in any of the patients. Histologically, our adrenal lymphangiomas showed a typical multicystic architecture with dilated spaces lined by

  13. Identification of immunohistochemical markers for distinguishing lung adenocarcinoma from squamous cell carcinoma

    PubMed Central

    Zhan, Cheng; Yan, Li; Wang, Lin; Sun, Yang; Wang, Xingxing; Lin, Zongwu; Zhang, Yongxing; Wang, Qun

    2015-01-01

    Background Immunohistochemical staining has been widely used in distinguishing lung adenocarcinoma (LUAD) from lung squamous cell carcinoma (LUSC), which is of vital importance for the diagnosis and treatment of lung cancer. Due to the lack of a comprehensive analysis of different lung cancer subtypes, there may still be undiscovered markers with higher diagnostic accuracy. Methods Herein first, we systematically analyzed high-throughput data obtained from The Cancer Genome Atlas (TCGA) database. Combining differently expressed gene screening and receiver operating characteristic (ROC) curve analysis, we attempted to identify the genes which might be suitable as immunohistochemical markers in distinguishing LUAD from LUSC. Then we detected the expression of six of these genes (MLPH, TMC5, SFTA3, DSG3, DSC3 and CALML3) in lung cancer sections using immunohistochemical staining. Results A number of genes were identified as candidate immunohistochemical markers with high sensitivity and specificity in distinguishing LUAD from LUSC. Then the staining results confirmed the potentials of the six genes (MLPH, TMC5, SFTA3, DSG3, DSC3 and CALML3) in distinguishing LUAD from LUSC, and their sensitivity and specificity were not less than many commonly used markers. Conclusions The results revealed that the six genes (MLPH, TMC5, SFTA3, DSG3, DSC3 and CALML3) might be suitable markers in distinguishing LUAD from LUSC, and also validated the feasibility of our methods for identification of candidate markers from high-throughput data. PMID:26380766

  14. [Prospects for using immunohistochemical methods in forensic medical thanatology].

    PubMed

    Bogomolov, D V; Bogomolova, I N; Karavaeva, I E

    2009-01-01

    This review of Russian and foreign literature is focused on the use of immunohistochemical methods in forensic medical practice. It shows that forensic medical specialists not infrequently underestimate the value of these techniques. Recommendations are proposed for a more extensive application of immunohistochemical methods in practical and fundamental medico-legal thanatology.

  15. Consensus statement on the immunohistochemical detection of ocular lymphatic vessels.

    PubMed

    Schroedl, Falk; Kaser-Eichberger, Alexandra; Schlereth, Simona L; Bock, Felix; Regenfuss, Birgit; Reitsamer, Herbert A; Lutty, Gerard A; Maruyama, Kazuichi; Chen, Lu; Lütjen-Drecoll, Elke; Dana, Reza; Kerjaschki, Dontscho; Alitalo, Kari; De Stefano, Maria Egle; Junghans, Barbara M; Heindl, Ludwig M; Cursiefen, Claus

    2014-10-01

    There is currently considerable controversy about existence and classification of "lymphatic vessels" in the eye. Some of the confusion is certainly caused by inappropriate use (or nonuse) of the correct immunohistochemical markers. Many experts in the field expressed the need for a consensus statement, and, in this perspective, authors offer arguments and solutions to reliably continue with immunohistochemical ocular lymphatic research.

  16. Prognostic value of immunohistochemical markers in malignant thymic epithelial tumors

    PubMed Central

    Leisibach, Priska; Schneiter, Didier; Soltermann, Alex; Yamada, Yoshi; Weder, Walter

    2016-01-01

    Background Thymic epithelial tumors (TET) are rare neoplasms with inconsistent treatment strategies. When researching for molecular pathways to find new therapies, the correlation between specific molecular markers and outcome has only rarely been investigated. The aim of this study was to investigate the correlation between survival, metastatic potential and invasiveness of aggressive subtypes of TET and immunohistochemical markers. Methods Overall survival (OS), disease-free survival (DFS), progression-free survival (PFS) and metastasis-free survival (MFS) of patients with WHO type B2/B3 mixed type thymoma (MT), thymoma type B3 (B3) and thymic carcinoma (TC), undergoing surgery [1998–2013] were determined. Tumor specimens were stained using a tissue microarray (TMA) (CD117, CD5, p63, p40, p21, p27, p53, Bcl-2, Ki67, podoplanin, synaptophysin, PTEN and Pax8). Invasive behavior of primary tumors and the presence of extrathoracic metastases were assessed. Results We found in 23 patients included into this study (four MT, ten B3, nine TC) that (I) p21 expression in the cytoplasm significantly correlated with a decrease of OS (P=0.016), PFS (P=0.034) and MFS (P=0.005); (II) MFS was significantly shorter when the combination of p21-low p27-low p53-high was present (P=0.029); and (III) nuclear p27 (P=0.042), Ki-67 (P=0.024) and podoplanin (P=0.05) expression correlated with the presence of extrathoracic metastases. Conclusions The main finding of this study is that cytoplasmic p21 expression negatively influences the outcome of malignant TETs and correlates with metastatic activity. Additionally, selected immunohistochemical markers correlate with the distant metastatic potential of TETs. These results may contribute to the stratification of diagnosis and improvement of treatment strategies for thymic malignancies. PMID:27747012

  17. HPV immunohistochemical testing and cervical dysplasia

    PubMed Central

    MUREŞAN, DANIEL; ROTAR, IOANA CRISTINA; APOSTOL, SILVANA; COROIU, GEORGIANA; STAMATIAN, FLORIN

    2016-01-01

    Background and aim HPV (Human Papilloma Virus) infection represents a necessary condition for cervical carcinogenesis. The purpose of this study was to evaluate the efficiency of HPV testing using an immunohistochemical staining kit with implications upon both diagnosis and treatment of cervical intraepithelial neoplasia (CIN). Methods Seventy-nine patients and eighty-six controls were enrolled in the study. Each patient had completed a physical examination, gynecological examination with cervical sampling using a liquid-based cytology system and also colposcopy. The cervical samples were analyzed according to Bethesda terminology and HPV-HR immunohistochemical staining was performed. In all the patients with high-grade lesion a surgical excision procedure was performed followed by pathological examination of the specimen. The collected data were analyzed using statistical software. Results The colposcopic examination has detected acetowhite modifications of the cervical epithelium in 47% of patients with ASC-US (Atypical squamous cells of undetermined significance) in 71% of patients with LSIL (Low grade squamous intraepithelial lesion) and in 100% of patients with HSIL ( High grade squamous intraepithelial lesion). The biopsy confirmed the diagnosis of LSIL in 27% of biopsy specimens in patients with ASC-US and in 79.99% of patients with LSIL respectively. In all patients with HSIL the diagnosis was CIN II or higher. The percentage of HPV-HR (Human Papilloma Virus – High Risk) positivity porportionaly increased with the severity of cytological diagnosis: 30% in ASC-US, 42.86% in LSIL and 75% in HSIL patients. The sensitivity of detection of HPV-HR was 50% with CI 95% [17.45;82.55] for ASC-US, 77.77% with CI 95% [51.91;92.62] for LSIL and 81.81% with CI 95% [58.99;94.00] for HSIL. Conclusion HPV testing can be an important screening tool for cervical dysplasia. The HPV testing targeting high risk types is indicated for ASC-US and LSIL triage. The present work

  18. Pancreatic neuroendocrine tumor and solid-pseudopapillary neoplasm: Key immunohistochemical profiles for differential diagnosis

    PubMed Central

    Ohara, Yusuke; Oda, Tatsuya; Hashimoto, Shinji; Akashi, Yoshimasa; Miyamoto, Ryoichi; Enomoto, Tsuyoshi; Satomi, Kaishi; Morishita, Yukio; Ohkohchi, Nobuhiro

    2016-01-01

    AIM To reveal better diagnostic markers for differentiating neuroendocrine tumor (NET) from solid-pseudopapillary neoplasm (SPN), focusing primarily on immunohistochemical analysis. METHODS We reviewed 30 pancreatic surgical specimens of NET (24 cases) and SPN (6 cases). We carried out comprehensive immunohistochemical profiling using 9 markers: Synaptophysin, chromogranin A, pan-cytokeratin, E-cadherin, progesterone receptor, vimentin, α-1-antitrypsin, CD10, and β-catenin. RESULTS E-cadherin staining in NETs, and nuclear labeling of β-catenin in SPNs were the most sensitive and specific markers. Dot-like staining of chromogranin A might indicate the possibility of SPNs rather than NETs. The other six markers were not useful because their expression overlapped widely between NETs and SPNs. Moreover, two cases that had been initially diagnosed as NETs on the basis of their morphological features, demonstrated SPN-like immunohistochemical profiles. Careful diagnosis is crucial as we actually found two confusing cases showing disagreement between the tumor morphology and immunohistochemical profiles. CONCLUSION E-cadherin, chromogranin A, and β-catenin were the most useful markers which should be employed for differentiating between NET and SPN.

  19. Neutron Activation Analysis of Water - A Review

    NASA Technical Reports Server (NTRS)

    Buchanan, John D.

    1971-01-01

    Recent developments in this field are emphasized. After a brief review of basic principles, topics discussed include sources of neutrons, pre-irradiation physical and chemical treatment of samples, neutron capture and gamma-ray analysis, and selected applications. Applications of neutron activation analysis of water have increased rapidly within the last few years and may be expected to increase in the future.

  20. Immunohistochemical diagnosis of eastern equine encephalomyelitis.

    PubMed

    Patterson, J S; Maes, R K; Mullaney, T P; Benson, C L

    1996-04-01

    An immunohistochemical (IHC) assay was developed for the detection of eastern equine encephalomyelitis (EEE) virus antigen in formalin-fixed, paraffin-embedded tissues. All cases of EEE diagnosed at the Michigan State University Animal Health Diagnostic Laboratory from 1991 through 1994 were evaluated. The diagnosis was based on histopathologic examination of the brain and confirmatory virus, isolation. Sections of cerebrum from 26 equids and 5 birds were assessed by IHC. Histologically normal brain tissues from 2 horses and 1 pheasant and brain tissues from 2 cases of equine neurologic disease with diagnoses other than EEE served as negative controls. The IHC assay was based on standard streptavidin-biotin technology, using a commercially available kit and a monospecific polyclonal primary antibody preparation derived from murine ascitic fluid. Nineteen of 20 equids and all 5 birds positive by histopathology and virus isolation were positive for EEE virus antigen by IHC. Three equids with histologic lesions compatible with a diagnosis of EEE but negative by virus isolation also were negative for virus antigen by IHC. In 3 other equids, histopathology and IHC were positive for EEE, but virus isolation was not attempted because of contamination of the brain specimen. The IHC assay of formalin-fixed, paraffin-embedded brain tissues for EEE virus antigen is a rapid, effective test for confirming a histopathologic diagnosis of EEE, and assay results correlate well with virus isolation results.

  1. Ultrastructural and immunohistochemical studies of rat epididymis.

    PubMed

    Francavilla, S; De Martino, C; Scorza Barcellona, P; Natali, P G

    1983-01-01

    The anatomical distribution of smooth muscle actin, myosin, fibronectin and basement membrane has been investigated immunohistochemically, using the indirect immunofluorescence technique, in the rat epididymis. The findings were correlated with the ultrastructural organization of the organ. Actin was found to be distributed in the stereociliary region of the epithelial principal cells and in the terminal web region. Actin was also visible along the base of the epithelium. Myosin was detected in the terminal web and in the terminal bar regions of the epithelium. The contractile cells showed a strong stain for both proteins. Basement membrane immunoreactivity was distributed along the epithelial basement membrane and around the contractile cells of the wall. In the cauda, between the epithelium and the contractile cell layers, the lamina propria, containing blood vessels and a thin layer of cells, was negative for all antigens investigated. Fibronectin showed a granular distribution around the contractile cells, mainly in the cauda. The ultrastructural study showed only thin (5-6 nm in diameter) filaments in the stereocilia and terminal web region. Thin filaments were also visible in the cytoplasm of the basal cells, thus suggesting a contractile function of this cell type. The heterogeneous appearance of the contractile cells of the wall seemed to support the different contractile pattern of the epididymal regions: caput, corpus and cauda. The cells present in the lamina propria showed cytoplasmic vesicles with dark granules resembling the "A" cell granules of the endocrine pancreas and gut mucosa cells.

  2. Ultrastructural and immunohistochemical studies of rat epididymis.

    PubMed

    Francavilla, S; De Martino, C; Scorza Barcellona, P; Natali, P G

    1983-01-01

    The anatomical distribution of smooth muscle actin, myosin, fibronectin and basement membrane has been investigated immunohistochemically, using the indirect immunofluorescence technique, in the rat epididymis. The findings were correlated with the ultrastructural organization of the organ. Actin was found to be distributed in the stereociliary region of the epithelial principal cells and in the terminal web region. Actin was also visible along the base of the epithelium. Myosin was detected in the terminal web and in the terminal bar regions of the epithelium. The contractile cells showed a strong stain for both proteins. Basement membrane immunoreactivity was distributed along the epithelial basement membrane and around the contractile cells of the wall. In the cauda, between the epithelium and the contractile cell layers, the lamina propria, containing blood vessels and a thin layer of cells, was negative for all antigens investigated. Fibronectin showed a granular distribution around the contractile cells, mainly in the cauda. The ultrastructural study showed only thin (5-6 nm in diameter) filaments in the stereocilia and terminal web region. Thin filaments were also visible in the cytoplasm of the basal cells, thus suggesting a contractile function of this cell type. The heterogeneous appearance of the contractile cells of the wall seemed to support the different contractile pattern of the epididymal regions: caput, corpus and cauda. The cells present in the lamina propria showed cytoplasmic vesicles with dark granules resembling the "A" cell granules of the endocrine pancreas and gut mucosa cells. PMID:6354463

  3. Immunohistochemical diagnosis of eastern equine encephalomyelitis.

    PubMed

    Patterson, J S; Maes, R K; Mullaney, T P; Benson, C L

    1996-04-01

    An immunohistochemical (IHC) assay was developed for the detection of eastern equine encephalomyelitis (EEE) virus antigen in formalin-fixed, paraffin-embedded tissues. All cases of EEE diagnosed at the Michigan State University Animal Health Diagnostic Laboratory from 1991 through 1994 were evaluated. The diagnosis was based on histopathologic examination of the brain and confirmatory virus, isolation. Sections of cerebrum from 26 equids and 5 birds were assessed by IHC. Histologically normal brain tissues from 2 horses and 1 pheasant and brain tissues from 2 cases of equine neurologic disease with diagnoses other than EEE served as negative controls. The IHC assay was based on standard streptavidin-biotin technology, using a commercially available kit and a monospecific polyclonal primary antibody preparation derived from murine ascitic fluid. Nineteen of 20 equids and all 5 birds positive by histopathology and virus isolation were positive for EEE virus antigen by IHC. Three equids with histologic lesions compatible with a diagnosis of EEE but negative by virus isolation also were negative for virus antigen by IHC. In 3 other equids, histopathology and IHC were positive for EEE, but virus isolation was not attempted because of contamination of the brain specimen. The IHC assay of formalin-fixed, paraffin-embedded brain tissues for EEE virus antigen is a rapid, effective test for confirming a histopathologic diagnosis of EEE, and assay results correlate well with virus isolation results. PMID:8744735

  4. Human hair neutron activation analysis: Analysis on population level, mapping

    NASA Astrophysics Data System (ADS)

    Zhuk, L. I.; Kist, A. A.

    1999-01-01

    Neutron activation analysis is an outstanding analytical method having very wide applications in various fields. Analysis of human hair within last decades mostly based on neutron activation analysis is a very attractive illustration of the application of nuclear analytical techniques. Very interesting question is how the elemental composition differs in different areas or cities. In this connection the present paper gives average data and maps of various localities in the vicinity of drying-out Aral Sea and of various industrial cities in Central Asia.

  5. An Immunohistochemical Algorithm for Ovarian Carcinoma Typing

    PubMed Central

    Rahimi, Kurosh; Rambau, Peter F.; Naugler, Christopher; Le Page, Cécile; Meunier, Liliane; de Ladurantaye, Manon; Lee, Sandra; Leung, Samuel; Goode, Ellen L.; Ramus, Susan J.; Carlson, Joseph W.; Li, Xiaodong; Ewanowich, Carol A.; Kelemen, Linda E.; Vanderhyden, Barbara; Provencher, Diane; Huntsman, David; Lee, Cheng-Han; Gilks, C. Blake; Mes Masson, Anne-Marie

    2016-01-01

    There are 5 major histotypes of ovarian carcinomas. Diagnostic typing criteria have evolved over time, and past cohorts may be misclassified by current standards. Our objective was to reclassify the recently assembled Canadian Ovarian Experimental Unified Resource and the Alberta Ovarian Tumor Type cohorts using immunohistochemical (IHC) biomarkers and to develop an IHC algorithm for ovarian carcinoma histotyping. A total of 1626 ovarian carcinoma samples from the Canadian Ovarian Experimental Unified Resource and the Alberta Ovarian Tumor Type were subjected to a reclassification by comparing the original with the predicted histotype. Histotype prediction was derived from a nominal logistic regression modeling using a previously reclassified cohort (N=784) with the binary input of 8 IHC markers. Cases with discordant original or predicted histotypes were subjected to arbitration. After reclassification, 1762 cases from all cohorts were subjected to prediction models (χ2 Automatic Interaction Detection, recursive partitioning, and nominal logistic regression) with a variable IHC marker input. The histologic type was confirmed in 1521/1626 (93.5%) cases of the Canadian Ovarian Experimental Unified Resource and the Alberta Ovarian Tumor Type cohorts. The highest misclassification occurred in the endometrioid type, where most of the changes involved reclassification from endometrioid to high-grade serous carcinoma, which was additionally supported by mutational data and outcome. Using the reclassified histotype as the endpoint, a 4-marker prediction model correctly classified 88%, a 6-marker 91%, and an 8-marker 93% of the 1762 cases. This study provides statistically validated, inexpensive IHC algorithms, which have versatile applications in research, clinical practice, and clinical trials. PMID:26974996

  6. [Immunohistochemical studies with middle ear mucosal remnants in cholesteatoma].

    PubMed

    Sudhoff, H; Borkowski, G; Bujia, J; Hildmann, H; Fisseler-Eckhoff, A

    1997-08-01

    The development of a middle ear cholesteatoma is usually associated with chronic inflammation and displacement of the mucosa present by the invading squamous epithelium. To analyze the clinically different behaviors of both epithelia, we used immunohistochemical methods to study the distribution and expression of interleukin-1 (Il-1), transforming growth factor-alpha (TGF-alpha), epidermal growth factor (EGF), epidermal growth factor-receptor (EGF-R), the proliferation marker MIB 1, c-myc proto-oncogene product and activation marker 4F2. Results stromal that keratinocytes in a cholesteatoma exhibited a much higher activation and proliferation rate when compared to middle ear mucosa cells. Middle ear epithelial cells showed no immunoreactivity for TGF-alpha, EGF-R, Il-1 and c-myc in contrast to the markedly positive immunoreactivity found in cholesteatoma matrix. The local release of cytokines and growth factors, such as TGF-alpha, EGF and Il-1 by inflammatory cells seems to be an important factor for the hyper-proliferative behavior of cholesteatoma epithelium. Our findings could contribute to the pathogenesis of middle ear cholesteatoma and give a possible explanation for the sustained progression of its growth leading to displacement of the middle ear mucosa.

  7. Comparative analysis of Napsin A, alpha-methylacyl-coenzyme A racemase (AMACR, P504S), and hepatocyte nuclear factor 1 beta as diagnostic markers of ovarian clear cell carcinoma: an immunohistochemical study of 279 ovarian tumours.

    PubMed

    Fadare, Oluwole; Zhao, Chengquan; Khabele, Dineo; Parkash, Vinita; Quick, Charles M; Gwin, Katja; Desouki, Mohamed M

    2015-02-01

    Napsin A and α-methylacyl-coenzyme A racemase (AMACR, P504S) have recently been described as being frequently expressed in clear cell carcinomas (CCC) of the gynecological tract. The present study was conducted to assess the test performance of these newer markers relative to the more traditional marker, hepatocyte nuclear factor 1β (HNF1β), in a large and histotypically diverse dataset. A total of 279 ovarian tumours in tissue microarrays were immunohistochemically assessed for the expression of Napsin A, AMACR and HNF1β. HNF1β, Napsin A and AMACR were expressed in 92%, 82% and 63% of 65 CCC, 7%, 1% and 1% of 101 serous carcinomas, 37%, 5.3% and 0% of 19 endometrioid carcinomas, 60%, 0% and 0% of 45 mucinous tumours, 100%, 0% and 0% of seven yolk sac tumours, and 0%, 16.7% and 16.7% of six steroid cell tumours NOS, respectively. All other tumours, including 18 adult-type granulosa cell tumours, eight dysgerminomas and nine other miscellaneous tumour types were negative for all three markers. Using a benchmark of ≥1% of tumour cells for positivity and CCC as the diagnostic end-point, the sensitivity, specificity, negative predictive value and positive predictive value of Napsin A expression were 0.82, 0.99, 0.94, and 0.98, respectively (odds ratio 439, p < 0.0001). Respective parameters were 0.92, 0.79, 0.97, and 0.58 (odds ratio 44, p < 0.0001) for HNF1β and 0.63, 0.99, 0.89, and 0.5 (odds ratio 112, p < 0.0001) for AMACR. The combination of any two positive markers, irrespective of the staining pattern of the third, significantly predicted the CCC histotype in every analytic scenario. In summary, HNF1β is highly sensitive but is suboptimally specific in isolation, whereas AMACR is highly specific but is suboptimally sensitive. Napsin A is specific but of intermediate sensitivity. Napsin A, AMACR and HNF1β are all viable markers of CCC that can be deployed as components of larger panels when CCC is a diagnostic consideration.

  8. Comparative analysis of Napsin A, alpha-methylacyl-coenzyme A racemase (AMACR, P504S), and hepatocyte nuclear factor 1 beta as diagnostic markers of ovarian clear cell carcinoma: an immunohistochemical study of 279 ovarian tumours.

    PubMed

    Fadare, Oluwole; Zhao, Chengquan; Khabele, Dineo; Parkash, Vinita; Quick, Charles M; Gwin, Katja; Desouki, Mohamed M

    2015-02-01

    Napsin A and α-methylacyl-coenzyme A racemase (AMACR, P504S) have recently been described as being frequently expressed in clear cell carcinomas (CCC) of the gynecological tract. The present study was conducted to assess the test performance of these newer markers relative to the more traditional marker, hepatocyte nuclear factor 1β (HNF1β), in a large and histotypically diverse dataset. A total of 279 ovarian tumours in tissue microarrays were immunohistochemically assessed for the expression of Napsin A, AMACR and HNF1β. HNF1β, Napsin A and AMACR were expressed in 92%, 82% and 63% of 65 CCC, 7%, 1% and 1% of 101 serous carcinomas, 37%, 5.3% and 0% of 19 endometrioid carcinomas, 60%, 0% and 0% of 45 mucinous tumours, 100%, 0% and 0% of seven yolk sac tumours, and 0%, 16.7% and 16.7% of six steroid cell tumours NOS, respectively. All other tumours, including 18 adult-type granulosa cell tumours, eight dysgerminomas and nine other miscellaneous tumour types were negative for all three markers. Using a benchmark of ≥1% of tumour cells for positivity and CCC as the diagnostic end-point, the sensitivity, specificity, negative predictive value and positive predictive value of Napsin A expression were 0.82, 0.99, 0.94, and 0.98, respectively (odds ratio 439, p < 0.0001). Respective parameters were 0.92, 0.79, 0.97, and 0.58 (odds ratio 44, p < 0.0001) for HNF1β and 0.63, 0.99, 0.89, and 0.5 (odds ratio 112, p < 0.0001) for AMACR. The combination of any two positive markers, irrespective of the staining pattern of the third, significantly predicted the CCC histotype in every analytic scenario. In summary, HNF1β is highly sensitive but is suboptimally specific in isolation, whereas AMACR is highly specific but is suboptimally sensitive. Napsin A is specific but of intermediate sensitivity. Napsin A, AMACR and HNF1β are all viable markers of CCC that can be deployed as components of larger panels when CCC is a diagnostic consideration. PMID:25551297

  9. Radioactivity analysis in niobium activation foils

    SciTech Connect

    Mueller, G.E.

    1995-06-01

    The motivation for this study was to measure and analyze the activity of six (6) niobium (Nb) foils (the x-rays from an internal transition in Nb-93m) and apply this information with previously obtained activation foil data. The niobium data was used to determine the epithermal to MeV range for the neutron spectrum and fluence. The foil activation data was re-evaluated in a spectrum analysis code (STAY`SL) to provide new estimates of the exposure at the Los Alamos Spallation Radiation Effect Facility (LASREF). The activity of the niobium foils was measured and analyzed at the University of Missouri-Columbia (UMC) under the direction of Professor William Miller. The spectrum analysis was performed at the University of Missouri-Rolla (UMR) by Professor Gary Mueller.

  10. Immunohistochemical study of pancreatic neuroendocrine tumor in Panthera tigris tigris.

    PubMed

    Nyska, A; Goldstein, J; Eshkar, G; Klein, B

    1996-07-01

    The histological and immunohistochemical characteristics of a case of pancreatic neuroendocrine tumor are described in a 14-yr-old female Bengal tiger (Panthera tigris tigris) housed at the New Biblical Zoo of Jerusalem, Jerusalem, Israel, 1994. The neoplastic cells were immunohistochemically negative for insulin and glucagon, slightly positive for neuron-specific enolase, moderately positive for serotonin and somatostatin, and markedly positive for chromogranine A and gastrin. This is the first documentation of a pancreatic neuroendocrine tumor in the tiger.

  11. Immunohistochemical study of pancreatic neuroendocrine tumor in Panthera tigris tigris.

    PubMed

    Nyska, A; Goldstein, J; Eshkar, G; Klein, B

    1996-07-01

    The histological and immunohistochemical characteristics of a case of pancreatic neuroendocrine tumor are described in a 14-yr-old female Bengal tiger (Panthera tigris tigris) housed at the New Biblical Zoo of Jerusalem, Jerusalem, Israel, 1994. The neoplastic cells were immunohistochemically negative for insulin and glucagon, slightly positive for neuron-specific enolase, moderately positive for serotonin and somatostatin, and markedly positive for chromogranine A and gastrin. This is the first documentation of a pancreatic neuroendocrine tumor in the tiger. PMID:8827685

  12. GABA distribution in the central vestibular system after retroauricular galvanic stimulation. An immunohistochemical study.

    PubMed

    Okami, K; Sekitani, T; Ogata, M; Matsuda, Y; Ogata, Y; Kanaya, K; Tahara, T

    1991-01-01

    The changes of the neurotransmitter (GABA) distribution in the brain stem of rats by retroauricular galvanic stimulation were investigated using the immunohistochemical method. In the lateral vestibular nucleus GABA-like immunoreactivity was more intensive on the side ipsilateral to the anodal stimulation than on the other side. It is concluded that retroauricular galvanic stimulation causes some changes in the inhibitory activity of the lateral vestibulo-spinal tract and of the spinal motor neuron.

  13. Magnetoreception in birds: I. Immunohistochemical studies concerning the cryptochrome cycle.

    PubMed

    Nießner, Christine; Denzau, Susanne; Peichl, Leo; Wiltschko, Wolfgang; Wiltschko, Roswitha

    2014-12-01

    Cryptochrome 1a, located in the UV/violet-sensitive cones in the avian retina, is discussed as receptor molecule for the magnetic compass of birds. Our previous immunohistochemical studies of chicken retinae with an antiserum that labelled only activated cryptochrome 1a had shown activation of cryptochrome 1a under 373 nm UV, 424 nm blue, 502 nm turquoise and 565 nm green light. Green light, however, does not allow the first step of photoreduction of oxidized cryptochromes to the semiquinone. As the chickens had been kept under 'white' light before, we suggested that there was a supply of the semiquinone present at the beginning of the exposure to green light, which could be further reduced and then re-oxidized. To test this hypothesis, we exposed chickens to various wavelengths (1) for 30 min after being kept in daylight, (2) for 30 min after a 30 min pre-exposure to total darkness, and (3) for 1 h after being kept in daylight. In the first case, we found activated cryptochrome 1a under UV, blue, turquoise and green light; in the second two cases we found activated cryptochrome 1a only under UV to turquoise light, where the complete redox cycle of cryptochrome can run, but not under green light. This observation is in agreement with the hypothesis that activated cryptochrome 1a is found as long as there is some of the semiquinone left, but not when the supply is depleted. It supports the idea that the crucial radical pair for magnetoreception is generated during re-oxidation.

  14. Magnetoreception in birds: I. Immunohistochemical studies concerning the cryptochrome cycle.

    PubMed

    Nießner, Christine; Denzau, Susanne; Peichl, Leo; Wiltschko, Wolfgang; Wiltschko, Roswitha

    2014-12-01

    Cryptochrome 1a, located in the UV/violet-sensitive cones in the avian retina, is discussed as receptor molecule for the magnetic compass of birds. Our previous immunohistochemical studies of chicken retinae with an antiserum that labelled only activated cryptochrome 1a had shown activation of cryptochrome 1a under 373 nm UV, 424 nm blue, 502 nm turquoise and 565 nm green light. Green light, however, does not allow the first step of photoreduction of oxidized cryptochromes to the semiquinone. As the chickens had been kept under 'white' light before, we suggested that there was a supply of the semiquinone present at the beginning of the exposure to green light, which could be further reduced and then re-oxidized. To test this hypothesis, we exposed chickens to various wavelengths (1) for 30 min after being kept in daylight, (2) for 30 min after a 30 min pre-exposure to total darkness, and (3) for 1 h after being kept in daylight. In the first case, we found activated cryptochrome 1a under UV, blue, turquoise and green light; in the second two cases we found activated cryptochrome 1a only under UV to turquoise light, where the complete redox cycle of cryptochrome can run, but not under green light. This observation is in agreement with the hypothesis that activated cryptochrome 1a is found as long as there is some of the semiquinone left, but not when the supply is depleted. It supports the idea that the crucial radical pair for magnetoreception is generated during re-oxidation. PMID:25472972

  15. Transforming Teacher Education, An Activity Theory Analysis

    ERIC Educational Resources Information Center

    McNicholl, Jane; Blake, Allan

    2013-01-01

    This paper explores the work of teacher education in England and Scotland. It seeks to locate this work within conflicting sociocultural views of professional practice and academic work. Drawing on an activity theory framework that integrates the analysis of these seemingly contradictory discourses with a study of teacher educators' practical…

  16. Computer-automated neutron activation analysis system

    SciTech Connect

    Minor, M.M.; Garcia, S.R.

    1983-01-01

    An automated delayed neutron counting and instrumental neutron activation analysis system has been developed at Los Alamos National Laboratory's Omega West Reactor (OWR) to analyze samples for uranium and 31 additional elements with a maximum throughput of 400 samples per day. 5 references.

  17. Immunohistochemical Characterization of Leukocytic Subpopulations in Chronic Endometritis

    PubMed Central

    Venuti, Susan; Brown, Sharla; Collins, Julie

    1996-01-01

    Objective: We analyzed the histologic and immunohistochemical changes in the endometrial leukocytic subpopulations to determine which of them are characteristic of chronic endometritis. Results: Endometrial biopsies from 25 cases of chronic endometritis and 35 controls were studied. Characteristic morphologic findings included the presence of a plasma cell infiltrate, and a prominent, albeit non-specific, lymphocytic infiltrate in all patients with endometritis. A neutrophilic infiltrate was also noted in 90% of the patients. Other non-specific histologic findings included occasional prominent lymphoid aggregates, stromal edema, hemorrhage, and necrosis and cystic dilation of some glands in endometria of patients with chronic endometritis. Endometrial immune cells were investigated immunohistochemically using antibodies to CD3 (pan-T), CD20 (pan-B, L26), and Ham-56 (macrophage). In control patients, endometrial immune cells were predominantly composed of CD3 and Ham-56 positive cells. CD20 positive cells comprised <2% of immune cells in control patients [mean: <2 cells/high power field (HPF)]. Large numbers of CD20 and CD3 lymphocytes were seen in endometria of patients with chronic endometritis. A semiquantitative analysis showed that the numbers of CD20 and CD3 positive cells in patients with chronic endometritis were increased 50- and 3-fold, respectively, when compared to those of control patients (mean B cells: 49 vs. 2 cells/HPF; mean T cells: 149 vs. 45 cells/HPF). CD20 positive cells were predominantly seen in subepithelial and periglandular aggregates. CD3 positive cells had a predominant stromal distribution and an occasional intra- or subepithelial localization. There was no difference in the number and distribution of Ham-56 positive cells in patients with or without endometritis. Conclusions: These findings suggest that CD20 cells may have a significant role in the pathogenesis of chronic endometritis and that immunostaining for B and T lymphocytes

  18. Intracholecystic papillary-tubular neoplasms (ICPN) of the gallbladder (neoplastic polyps, adenomas, and papillary neoplasms that are ≥1.0 cm): clinicopathologic and immunohistochemical analysis of 123 cases.

    PubMed

    Adsay, Volkan; Jang, Kee-Taek; Roa, Juan Carlos; Dursun, Nevra; Ohike, Nobuyuki; Bagci, Pelin; Basturk, Olca; Bandyopadhyay, Sudeshna; Cheng, Jeanette D; Sarmiento, Juan M; Escalona, Oscar Tapia; Goodman, Michael; Kong, So Yeon; Terry, Paul

    2012-09-01

    The literature on the clinicopathologic characteristics of tumoral intraepithelial neoplasms (neoplastic polyps) of the gallbladder (GB) is fairly limited, due in part to the variability in definition and terminology. Most reported adenomas (pyloric gland type and others) were microscopic and thus regarded as clinically inconsequential, whereas papillary in situ carcinomas have been largely considered a type of invasive adenocarcinoma under the heading of "papillary adenocarcinomas." In this study, 123 GB cases that have a well-defined exophytic preinvasive neoplasm measuring ≥1 cm were analyzed. The patients were predominantly female (F/M=2:1) with a mean age of 61 y and a median tumor size of 2.2 cm. Half of the patients presented with pain, and in the other half the neoplasm was detected incidentally. Other neoplasms, most being gastrointestinal tract malignancies, were present in 22% of cases. Gallstones were identified in only 20% of cases. Radiologically, almost half were diagnosed as "cancer," roughly half with polypoid tumor, and in 10% the lesion was missed. Pathologic findings: (1) The predominant configuration was papillary in 43%, tubulopapillary in 31%, tubular in 26%. (2) Each case was assigned a final lineage type on the basis of the predominant pattern (>75% of the lesion) on morphology, and supported with specific immunohistochemical cell lineage markers. The predominant cell lineage could be identified as biliary in 50% (66% of which were MUC1), gastric foveolar in 16% (all were MUC5AC), gastric pyloric in 20% (92% MUC6), intestinal in 8% (100% CK20; 75% CDX2; 50%, MUC2), and oncocytic in 6% (17% HepPar and 17% MUC6); however, 90% of cases had some amount of secondary or unclassifiable pattern and hybrid immunophenotypes. (3) Of the cases that would have qualified as "pyloric gland adenoma," 21/24 (88%) had at least focal high-grade dysplasia and 18% had associated invasive carcinoma. Conversely, 8 of 47 "papillary adenocarcinoma"-type cases

  19. Sonic hedgehog in oral squamous cell carcinoma: An immunohistochemical study

    PubMed Central

    Srinath, Sahana; Iyengar, Asha R; Mysorekar, Vijaya

    2016-01-01

    Background: Recent studies have revealed the involvement of hedgehog (Hh) signaling component in proliferation and invasive behavior of many carcinomas. Aim: This study aims to identify the expression of sonic Hh (SHH) protein of SHH pathway in oral epithelial dysplasia and oral squamous cell carcinoma (OSCC) using SHH (H-160) (Santa Cruz, sc-9042) which could have therapeutic implication in future. Materials and Methods: A total of 250 cases comprising 50 normal oral mucosa, 50 cases of oral epithelial dysplasia, 50 well, 50 moderate and 50 poorly differentiated OSCCs were included in the study. Immunohistochemical evaluation of SHH protein expression was conducted using monoclonal antibody. Interpretation of the expression was done by immunoreactive score of Remmele and Stegner (IRS) scoring method. Statistical Analysis: Chi-Square test was used to analyze the results. Results: The study showed that SHH signaling molecules are highly expressed in OSCC, and their expression was mainly in the cytoplasm of epithelial cells. Conclusion: The SHH signaling component is associated with the pathological parameter in OSCC and oral epithelial dysplasia. PMID:27721600

  20. Tenascin-C in chronic canine hepatitis: immunohistochemical localization and correlation with necro-inflammatory activity, fibrotic stage, and expression of alpha-smooth muscle actin, cytokeratin 7, and CD3+ cells.

    PubMed

    Mekonnen, G A; Ijzer, J; Nederbragt, H

    2007-11-01

    During fibrosis, the extracellular matrix (ECM) is continuously remodeled and increases in volume due to the production of various proteins. We studied the distribution of tenascin-C (TN-C) and the correlation of TN-C with the necro-inflammatory activity and expression of alpha-smooth muscle actin (alpha-SMA), cytokeratin 7 (CK7), and CD3+ T-lymphocytes in canine chronic hepatitis. This was analyzed using immunohistochemistry and semiquantitative scoring. We used 3 groups (n = 19) of dogs: group 1 (n = 5) with neonatal hepatitis/lobular dissecting hepatitis (NH/LDH), group 2 (n = 8) with chronic hepatitis/cirrhosis (CH/CIRR), and group 3 (n = 6) consisting of healthy animals. In normal livers, TN-C was localized in Disse's space and around bile ducts and blood vessels. In CH/CIRR livers, TN-C was localized at the periphery of the regenerating nodules and was conspicuous in the bridging fibrous bands. In NH/LDH, TN-C was diffusely distributed along the reticular fibers that dissected between single cells or groups of hepatocytes. alpha-SMA in the normal hepatic parenchyma showed an irregular distribution along the perisinusoidal linings. In other groups, alpha-SMA was increased in fibrotic septa and perisinusoidal linings. In normal livers, CK7 was positive in bile ducts. In other groups, CK7-expressing cells were conspicuous in the portal-parenchymal interface, the periphery of the regenerative nodules, and the degenerated parenchyma. The pattern of CD3+ lymphocytes was inversely proportional to that of TN-C. These results also showed that TN-C is strongly correlated with increased fibrotic stage, inflammatory activity, and expression of CK7 and alpha-SMA. TN-C, CK7, and CD3 expression did not differ between diagnostic groups.

  1. Enzyme catalysis enhanced dark-field imaging as a novel immunohistochemical method.

    PubMed

    Fan, Lin; Tian, Yanyan; Yin, Rong; Lou, Doudou; Zhang, Xizhi; Wang, Meng; Ma, Ming; Luo, Shouhua; Li, Suyi; Gu, Ning; Zhang, Yu

    2016-04-28

    Conventional immunohistochemistry is limited to subjective judgment based on human experience and thus it is clinically required to develop a quantitative immunohistochemical detection. 3,3'-Diaminobenzidin (DAB) aggregates, a type of staining product formed by conventional immunohistochemistry, were found to have a special optical property of dark-field imaging for the first time, and the mechanism was explored. On this basis, a novel immunohistochemical method based on dark-field imaging for detecting HER2 overexpressed in breast cancer was established, and the quantitative analysis standard and relevant software for measuring the scattering intensity was developed. In order to achieve a more sensitive detection, the HRP (horseradish peroxidase)-labeled secondary antibodies conjugated gold nanoparticles were constructed as nanoprobes to load more HRP enzymes, resulting in an enhanced DAB deposition as a dark-field label. Simultaneously, gold nanoparticles also act as a synergistically enhanced agent due to their mimicry of enzyme catalysis and dark-field scattering properties. PMID:26786242

  2. Immunohistochemical expression of types I and III collagen antibodies in the temporomandibular joint disc of human foetuses

    PubMed Central

    de Moraes, L.O.C.; Lodi, F.R.; Gomes, T.S.; Marques, S.R.; Oshima, C.T.F.; Lancellotti, C.L.P.; Rodríguez-Vázquez, J.F.; Mérida-Velasco, J.R.; Alonso, L.G.

    2011-01-01

    The objective was to study the morphology of the articular disc and analyse the immunohistochemical expression of types I and III collagen markers in the temporomandibular joint (TMJ) disc of human foetuses of different gestational ages. Twenty TMJ from human foetuses supplied by Universidade Federal de Uberaba with gestational ages from 17 to 24 weeks were studied. The gestational age of the foetuses was determined by measuring the crown-rump (CR) length. Macroscopically, the foetuses were fixed in 10% formalin solution and dissected by removing the skin and subcutaneous tissue and exposing the deep structures. Immunohistochemical markers of type I and III were used to characterize the existence of collagen fibres. Analysis of the immunohistochemical markers of types I and III collagen revealed the presence of heterotypical fibril networks. PMID:22073371

  3. Immunohistochemical expression of types I and III collagen antibodies in the temporomandibular joint disc of human foetuses.

    PubMed

    de Moraes, L O C; Lodi, F R; Gomes, T S; Marques, S R; Oshima, C T F; Lancellotti, C L P; Rodríguez-Vázquez, J F; Mérida-Velasco, J R; Alonso, L G

    2011-01-01

    The objective was to study the morphology of the articular disc and analyse the immunohistochemical expression of types I and III collagen markers in the temporomandibular joint (TMJ) disc of human foetuses of different gestational ages. Twenty TMJ from human foetuses supplied by Universidade Federal de Uberaba with gestational ages from 17 to 24 weeks were studied. The gestational age of the foetuses was determined by measuring the crown-rump (CR) length. Macroscopically, the foetuses were fixed in 10% formalin solution and dissected by removing the skin and subcutaneous tissue and exposing the deep structures. Immunohistochemical markers of type I and III were used to characterize the existence of collagen fibres. Analysis of the immunohistochemical markers of types I and III collagen revealed the presence of heterotypical fibril networks.

  4. Neutron activation analysis of Etruscan pottery

    SciTech Connect

    Whitehead, J.; Silverman, A.; Ouellet, C.G.; Clark, D.D.; Hossain, T.Z

    1992-07-01

    Neutron activation analysis (NAA) has been widely used in archaeology for compositional analysis of pottery samples taken from sites of archaeological importance. Elemental profiles can determine the place of manufacture. At Cornell, samples from an Etruscan site near Siena, Italy, are being studied. The goal of this study is to compile a trace element concentration profile for a large number of samples. These profiles will be matched with an existing data bank in an attempt to understand the place of origin for these samples. The 500 kW TRIGA reactor at the Ward Laboratory is used to collect NAA data for these samples. Experiments were done to set a procedure for the neutron activation analysis with respect to sample preparation, selection of irradiation container, definition of activation and counting parameters and data reduction. Currently, we are able to analyze some 27 elements in samples of mass 500 mg with a single irradiation of 4 hours and two sequences of counting. Our sensitivity for many of the trace elements is better than 1 ppm by weight under the conditions chosen. In this talk, details of our procedure, including quality assurance as measured by NIST standard reference materials, will be discussed. In addition, preliminary results from data treatment using cluster analysis will be presented. (author)

  5. Activity Analysis and Cost Analysis in Medical Schools.

    ERIC Educational Resources Information Center

    Koehler, John E.; Slighton, Robert L.

    There is no unique answer to the question of what an ongoing program costs in medical schools. The estimates of program costs generated by classical methods of cost accounting are unsatisfactory because such accounting cannot deal with the joint production or joint cost problem. Activity analysis models aim at calculating the impact of alternative…

  6. Histopathologic spectrum and immunohistochemical diagnosis of amebic meningoencephalitis.

    PubMed

    Guarner, Jeannette; Bartlett, Jeanine; Shieh, Wun-Ju; Paddock, Christopher D; Visvesvara, Govinda S; Zaki, Sherif R

    2007-12-01

    Traditionally, Naegleria fowleri infections are labeled primary amebic meningoencephalitis because of prominent meningeal neutrophilic inflammation. Acanthamoeba spp. and Balamuthia mandrillaris are labeled granulomatous amebic encephalitis because of parenchymal granulomatous inflammation. We compared histopathologic and immunohistochemical features of 18 cases with central nervous system free-living ameba infections. Immunohistochemical assays using polyclonal antibodies that reacted specifically against each genus identified 11 patients with Balamuthia infection, four with N. fowleri, and three with Acanthamoeba. Immunohistochemical assays highlighted the presence of trophozoites that were difficult to identify with hematoxylin and eosin stains in areas of necrosis or where macrophages were abundant. Immunohistochemical assays also demonstrated the presence of granular antigens inside macrophages and blood vessel walls. Amebic cysts were observed in three patients with Acanthamoeba infection and in three with Balamuthia. Patients with Acanthamoeba infection showed granulomatous inflammation. Patients with Naegleria infection had neutrophilic inflammation. Balamuthia infections showed a spectrum of inflammation that ranged from primarily neutrophils to granulomas. Meningitis was present in 88% of cases. Immunohistochemical assays were useful to demonstrate the presence of granular antigens and confirmed the genus of the ameba. The spectrum of inflammation in cases of Balamuthia meningoencephalitis is broader than previously described. The term amebic meningoencephalitis describes better the histopathologic findings than the currently used classification of primary amebic meningoencephalitis and granulomatous amebic encephalitis. PMID:17932496

  7. Histopathologic spectrum and immunohistochemical diagnosis of amebic meningoencephalitis.

    PubMed

    Guarner, Jeannette; Bartlett, Jeanine; Shieh, Wun-Ju; Paddock, Christopher D; Visvesvara, Govinda S; Zaki, Sherif R

    2007-12-01

    Traditionally, Naegleria fowleri infections are labeled primary amebic meningoencephalitis because of prominent meningeal neutrophilic inflammation. Acanthamoeba spp. and Balamuthia mandrillaris are labeled granulomatous amebic encephalitis because of parenchymal granulomatous inflammation. We compared histopathologic and immunohistochemical features of 18 cases with central nervous system free-living ameba infections. Immunohistochemical assays using polyclonal antibodies that reacted specifically against each genus identified 11 patients with Balamuthia infection, four with N. fowleri, and three with Acanthamoeba. Immunohistochemical assays highlighted the presence of trophozoites that were difficult to identify with hematoxylin and eosin stains in areas of necrosis or where macrophages were abundant. Immunohistochemical assays also demonstrated the presence of granular antigens inside macrophages and blood vessel walls. Amebic cysts were observed in three patients with Acanthamoeba infection and in three with Balamuthia. Patients with Acanthamoeba infection showed granulomatous inflammation. Patients with Naegleria infection had neutrophilic inflammation. Balamuthia infections showed a spectrum of inflammation that ranged from primarily neutrophils to granulomas. Meningitis was present in 88% of cases. Immunohistochemical assays were useful to demonstrate the presence of granular antigens and confirmed the genus of the ameba. The spectrum of inflammation in cases of Balamuthia meningoencephalitis is broader than previously described. The term amebic meningoencephalitis describes better the histopathologic findings than the currently used classification of primary amebic meningoencephalitis and granulomatous amebic encephalitis.

  8. MAGI-2 in prostate cancer: an immunohistochemical study.

    PubMed

    Goldstein, Jeffery; Borowsky, Alexander D; Goyal, Rajen; Roland, Joseph T; Arnold, Shanna A; Gellert, Lan L; Clark, Peter E; Hameed, Omar; Giannico, Giovanna A

    2016-06-01

    Membrane-associated guanylate kinase, WW and PDZ domain-containing protein 2 (MAGI-2) is a scaffolding protein that links cell adhesion molecules, receptors, and signaling molecules to the cytoskeleton and maintains the architecture of cell junctions. MAGI-2 gene rearrangements have recently been described in prostate cancer. We studied the immunohistochemical expression of MAGI-2 protein in prostate tissue. Seventy-eight radical prostatectomies were used to construct 3 tissue microarrays consisting of 512 cores, including benign tissue, benign prostatic hyperplasia, high-grade prostatic intraepithelial neoplasia (HGPIN), and adenocarcinoma, Gleason patterns 3 to 5. Immunohistochemistry for phosphatase and tensin homologue (PTEN) and double-stain MAGI-2/p63 was performed and analyzed by visual and image analysis, the latter as percent of analyzed area (%AREA), and mean optical density multiplied by %AREA (STAIN). By visual and image analysis, MAGI-2 was significantly higher in adenocarcinoma and HGPIN compared with benign (benign versus HGPIN P < .001; benign versus adenocarcinoma, P < .001). HGPIN and adenocarcinoma did not significantly differ by either modality. Using visual intensity to distinguish benign tissue and adenocarcinoma, a receiver operating curve yielded an area under the curve of 0.902. A STAIN threshold of 1470 yielded a sensitivity of 0.66 and specificity of 0.96. There was a significant correlation between PTEN and MAGI-2 staining for normal and benign prostatic hyperplasia, but this was lost in HGPIN and cancer. We conclude that MAGI-2 immunoreactivity is elevated in prostate cancer and HGPIN compared with normal tissue, and suggest that MAGI-2 may contribute to prostate carcinogenesis. This is the first report of MAGI-2 staining by immunohistochemistry in prostate cancer.

  9. Clinical significance of immunohistochemically detected extracellular matrix proteins and their spatial distribution in primary cancer.

    PubMed

    Čunderlíková, B

    2016-09-01

    Our understanding of cancer has evolved mainly from results of studies utilizing experimental models. Simplification inherent to in vitro cell culture models enabled potential ways of cell behaviour in response to various external stimuli to be described, but it has led also to disappointments in clinical trials, presumably due to the lack of crucial tissue components, including extracellular matrix (ECM). ECM and its role in healthy and diseased tissues are being explored extensively and significance of ECM for cell behaviour has been evidenced experimentally. Part of the information gathered in such research that is relevant for natural conditions of a human body can be identified by carefully designed analyses of human tissue samples. This review summarizes published information on clinical significance of ECM in cancer and examines whether effects of ECM on cell behaviour evidenced in vitro, could be supported by clinically based data acquired from analysis of tissue samples. Based on current approaches of clinical immunohistochemical analyses, impact of ECM components on tumour cell behaviour is vague. Except of traditionally considered limitations, other reasons may include lack of stratification of analyzed cases based on clinicopathologic parameters, inclusion of patients treated postoperatively by different treatments or neglecting complexity of interactions among tumour constituents. Nevertheless, reliable immunohistochemical studies represent a source of crucial information for design of tumour models comprising ECM corresponding to real clinical situation. Knowledge gathered from such immunohistochemical studies combined with achievements in tissue engineering hold promise for reversal of the unfavourable trends in the current translational oncologic research. PMID:27443915

  10. A possible complementary tool for diagnosing tuberculosis: a feasibility test of immunohistochemical markers.

    PubMed

    Seo, Kyung-Jin; Yoo, Chang-Young; Im, So-Young; Yeo, Chang-Dong; Jung, Ji-Han; Choi, Hyun-Ju; Yoo, Jin-Young

    2015-01-01

    Differentiation of tuberculous granuloma (TG) from non-tuberculous granuloma (NG) is histopathologically difficult. We evaluated the usefulness of selected immunohistochemical markers to differentiate tuberculous granuloma (TG) and non-tuberculous granuloma (NG). We selected six biomarkers (FoxP3, TNF-beta, E-selectin [ESEL], indoleamine 2,3-dioxygenase [IDO], lactoferrin [LACT], and tartrate-resistant acid phosphatase [TRAP]) and immunohistochemically analyzed their expression in the presence of two types of granulomatous tissue samples, TG (n = 36) and NG (n = 31), using a microarray format. Three of those six biomarkers (LACT, IDO, and TNF-beta) were moderately accurate in discriminating TG from NG, individually and in combination, according to ROC analysis (AUC = 0.7-0.89, sensitivity = 55.6-77.8%, specificity = 71.0-100%). Our data indicate that selected immunohistochemical markers (LACT, IDO, and TNF-beta) can be used in ancillary tests to differentiate TG from NG in tissue samples. Further large-scale studies are required to validate our results. PMID:26823702

  11. Immunohistochemical evaluation of experimental Vagococcus salmoninarum infection in rainbow trout (Oncorhynchus mykiss, Walbaum 1792).

    PubMed

    Yardimci, B; Didinen, B I; Onuk, E E; Metin, S; Ciftci, A; Kubilay, A; Pekmezci, G Z; Eralp, H

    2016-05-01

    The aim of this study was to investigate the pathogenesis and histopathological and immunohistochemical findings in rainbow trout (Oncorhynchus mykiss) following experimental vagococcosis. For this purpose, 60 rainbow trout were used. The experimental study used the pathogen Vagococcus salmoninarum. The fish were intraperitoneally (IP) administered with an inoculate containing 0.1 mL of the bacteria, resulting in a dose of 1.2 × 10(9) cfu mL(-1) per fish. For histopathological observations, tissue samples were taken from fish that died during the experiment and fish that survived until the end of the trial (60th day). All the tissue samples were immunohistochemically stained by the avidin-biotin-peroxidase complex and immunofluorescence methods using polyclonal antibody to detect V. salmoninarum antigens. In immunoperoxidase staining, positive reactions to bacterial antigens were most commonly seen in the kidney, heart and liver. In the immunofluorescence analysis, the distribution of antigens in the tissue and organs was similar to that observed with the immunoperoxidase staining. The results reveal an important correlation between histochemical and immunohistochemical staining in demonstrating the distribution of V. salmoninarum antigens in the affected tissues.

  12. Immunohistochemical expression pattern of MMR protein can specifically identify patients with colorectal cancer microsatellite instability.

    PubMed

    Amira, Arfaoui Toumi; Mouna, Trabelsi; Ahlem, Blel; Raoudha, Aloui; Majid, Ben Hmida; Amel, Hamza; Rachida, Zermani; Nadia, Kourdaa

    2014-07-01

    The microsatellite instability (MSI) pathway is found in most cases of hereditary nonpolyposis colorectal cancer (HNPCC) and in 12 % of sporadic colorectal cancer (CRC). It involves inactivation of deoxyribonucleic acid mismatch repair (MMR) genes MLH1, MSH2, PMS2, and MSH6. MMR germline mutation detections are an important supplement to HNPCC clinical diagnosis. It enables at-risk and mutation-positive relatives to be informed about their cancer risks and to benefit from intensive surveillance programs that have been proven to reduce the incidence of CRC. In this study, we analyzed for the first time in Tunisia the potential value of immunohistochemical assessment of MMR protein to identify microsatellite instability in CRC. We evaluate by immunohistochemistry MMR protein expression loss in tumoral tissue compared to positive expression in normal mucosa. Immunohistochemistry revealed loss of expression for MLH1, MSH2, MSH6, and PMS2 in 15, 21, 13, and 15 % of cases, respectively. Here, we report a more elevated frequency of MSI compared to data of the literature. In fact, by immunohistochemistry, 70 % of cases were shown to be MSS phenotype, whereas 30 % of cases, in our set, were instable. Moreover, according to molecular investigation, 71 % of cases were instable (MSI-H) and remaining cases were stable (29 %). Thus, we found a perfect association between MMR immunohistochemical analyses and MSI molecular investigation. Immunohistochemical analysis of MMR gene product expression may allow one to specifically identify MSI phenotype of patients with colorectal carcinomas.

  13. Comparative Sensitivity Analysis of Muscle Activation Dynamics.

    PubMed

    Rockenfeller, Robert; Günther, Michael; Schmitt, Syn; Götz, Thomas

    2015-01-01

    We mathematically compared two models of mammalian striated muscle activation dynamics proposed by Hatze and Zajac. Both models are representative for a broad variety of biomechanical models formulated as ordinary differential equations (ODEs). These models incorporate parameters that directly represent known physiological properties. Other parameters have been introduced to reproduce empirical observations. We used sensitivity analysis to investigate the influence of model parameters on the ODE solutions. In addition, we expanded an existing approach to treating initial conditions as parameters and to calculating second-order sensitivities. Furthermore, we used a global sensitivity analysis approach to include finite ranges of parameter values. Hence, a theoretician striving for model reduction could use the method for identifying particularly low sensitivities to detect superfluous parameters. An experimenter could use it for identifying particularly high sensitivities to improve parameter estimation. Hatze's nonlinear model incorporates some parameters to which activation dynamics is clearly more sensitive than to any parameter in Zajac's linear model. Other than Zajac's model, Hatze's model can, however, reproduce measured shifts in optimal muscle length with varied muscle activity. Accordingly we extracted a specific parameter set for Hatze's model that combines best with a particular muscle force-length relation. PMID:26417379

  14. Neutron activation analysis in archaeological chemistry

    SciTech Connect

    Harbottle, G.

    1987-01-01

    Neutron activation analysis has proven to be a convenient way of performing the chemical analysis of archaeologically-excavated artifacts and materials. It is fast and does not require tedious laboratory operations. It is multielement, sensitive, and can be made nondestructive. Neutron activation analysis in its instrumental form, i.e., involving no chemical separation, is ideally suited to automation and conveniently takes the first step in data flow patterns that are appropriate for many taxonomic and statistical operations. The future will doubtless see improvements in the practice of NAA in general, but in connection with archaeological science the greatest change will be the filling, interchange and widespread use of data banks based on compilations of analytical data. Since provenience-oriented data banks deal with materials (obsidian, ceramics, metals, semiprecious stones, building materials and sculptural media) that participated in trade networks, the analytical data is certain to be of interest to a rather broad group of archaeologists. It is to meet the needs of the whole archaeological community that archaeological chemistry must now turn.

  15. Perforin expression in plaque psoriasis: an immunohistochemical study.

    PubMed

    Samaka, Rehab Monir; Gaber, Mohamed A; Metwe, Nermin A

    2015-04-01

    Psoriasis (PsO) is T-cell-mediated disease resulting from aberrant activation of both innate and adaptive immunity. Perforin is a multi-domain, pore-forming protein. It is located within the cytoplasm of CD 8 cytotoxic T cells (CTLs) and natural killer cells (NK). The aim of this study was to evaluate the immunohistochemical (IHC) expression of perforin in lesional and perilesional skin of chronic plaque psoriatic patient and correlate its expression with the standard clinico-pathological variables. This prospective case-control study was conducted on 50 PsO patients and 30 age- and gender-matched healthy subjects as a control group. There were high-significant differences between lesional and perilesional skin of plaque PsO patients as regards to IHC perforin status and localization (p < 0.001 for both). There was a high-significant difference between positive and negative perforin cases as regards to psoriasis area severity index (PASI) (p < 0.000). There were significant differences between mild and moderate-to-severe intensity of IHC perforin expression as regards to triggering factors and PASI (p = 0.02 and 0.03, respectively). Localization of IHC perforin positive lymphocytes in both epidermis and dermis was significantly associated with higher degree of acanthosis and higher degree of inflammatory infiltrates in comparison with positive cells located in dermis (p = 0.001 for both). Perforin might have a putative signaling in early and late plaque PsO. Plaque psoriatic patients with positive perforin expression could be a candidate for a future target therapy to stop the proposed scenario and achieve a therapeutic response.

  16. Helicobacter pylori in Cholecystectomy Specimens-Morphological and Immunohistochemical Assessment

    PubMed Central

    Reddy, Venkatarami; Jena, Amitabh; Gavini, Siva; Thota, Asha; Nandyala, Rukamangadha; Chowhan, Amit Kumar

    2016-01-01

    Introduction Helicobacter pylori (H.pylori) is associated with gastritis, peptic ulcer, gastric carcinoma and gastric lymphoma. Current literature describes presence of H.pylori in various extra-gastric locations and its association with many diseases. Apart from the conventional location of gastric and duodenal mucosa, H.pylori have been isolated and cultured from gallbladder. Aim Analysis of cholecystectomy specimens to detect H.pylori by means of immunohistochemical staining. Materials and Methods There were a total of 118 cholecystectomy specimens received in the Department of Pathology in three months duration. We have performed immunostaining for H.pylori in 45 consecutive cases of cholecystectomy specimen. Clinical and other investigational information were retrieved from the medical records department. For each case, routine Haematoxylin and Eosin stain was studied. Immunohistochemistry (IHC) was done using purified polyclonal Helicobacter pylori antiserum. Results Majority of the patients had undergone laparoscopic cholecystectomy for the presenting complaint of right hypochondrial pain. Multiple pigmented stones were present in majority (27/45) of them. Immunostain for H.pylori was positive in ten cases. Six of these cases had pigmented gall stones, two had stones not specified and in two of the cases there were no stones. Conclusion Helicobacter pylori is present in gall bladder and is commonly seen in association with stones. A more detailed study of cholecystectomy cases (both neoplastic and non-neoplastic) with serological, culture and molecular data of H.pylori is desirable to study the pathogenesis of cholecystitis, its association with gall stones and other gall bladder disorders. PMID:27437221

  17. Analysis of DOE international environmental management activities

    SciTech Connect

    Ragaini, R.C.

    1995-09-01

    The Department of Energy`s (DOE) Strategic Plan (April 1994) states that DOE`s long-term vision includes world leadership in environmental restoration and waste management activities. The activities of the DOE Office of Environmental Management (EM) can play a key role in DOE`s goals of maintaining U.S. global competitiveness and ensuring the continuation of a world class science and technology community. DOE`s interest in attaining these goals stems partly from its participation in organizations like the Trade Policy Coordinating Committee (TPCC), with its National Environmental Export Promotion Strategy, which seeks to strengthen U.S. competitiveness and the building of public-private partnerships as part of U.S. industrial policy. The International Interactions Field Office task will build a communication network which will facilitate the efficient and effective communication between DOE Headquarters, Field Offices, and contractors. Under this network, Headquarters will provide the Field Offices with information on the Administration`s policies and activities (such as the DOE Strategic Plan), interagency activities, as well as relevant information from other field offices. Lawrence Livermore National Laboratory (LLNL) will, in turn, provide Headquarters with information on various international activities which, when appropriate, will be included in reports to groups like the TPCC and the EM Focus Areas. This task provides for the collection, review, and analysis of information on the more significant international environmental restoration and waste management initiatives and activities which have been used or are being considered at LLNL. Information gathering will focus on efforts and accomplishments in meeting the challenges of providing timely and cost effective cleanup of its environmentally damaged sites and facilities, especially through international technical exchanges and/or the implementation of foreign-development technologies.

  18. Scalable histopathological image analysis via active learning.

    PubMed

    Zhu, Yan; Zhang, Shaoting; Liu, Wei; Metaxas, Dimitris N

    2014-01-01

    Training an effective and scalable system for medical image analysis usually requires a large amount of labeled data, which incurs a tremendous annotation burden for pathologists. Recent progress in active learning can alleviate this issue, leading to a great reduction on the labeling cost without sacrificing the predicting accuracy too much. However, most existing active learning methods disregard the "structured information" that may exist in medical images (e.g., data from individual patients), and make a simplifying assumption that unlabeled data is independently and identically distributed. Both may not be suitable for real-world medical images. In this paper, we propose a novel batch-mode active learning method which explores and leverages such structured information in annotations of medical images to enforce diversity among the selected data, therefore maximizing the information gain. We formulate the active learning problem as an adaptive submodular function maximization problem subject to a partition matroid constraint, and further present an efficient greedy algorithm to achieve a good solution with a theoretically proven bound. We demonstrate the efficacy of our algorithm on thousands of histopathological images of breast microscopic tissues. PMID:25320821

  19. Evaluation of a direct, rapid immunohistochemical test for rabies diagnosis.

    PubMed

    Lembo, Tiziana; Niezgoda, Michael; Velasco-Villa, Andrés; Cleaveland, Sarah; Ernest, Eblate; Rupprecht, Charles E

    2006-02-01

    A direct rapid immunohistochemical test (dRIT) was evaluated under field and laboratory conditions to detect rabies virus antigen in frozen and glycerol-preserved field brain samples from northwestern Tanzania. Compared to the direct fluorescent antibody test, the traditional standard in rabies diagnosis, the dRIT was 100% sensitive and specific.

  20. Evaluation of a Direct, Rapid Immunohistochemical Test for Rabies Diagnosis

    PubMed Central

    Lembo, Tiziana; Velasco-Villa, Andrés; Cleaveland, Sarah; Ernest, Eblate; Rupprecht, Charles E.

    2006-01-01

    A direct rapid immunohistochemical test (dRIT) was evaluated under field and laboratory conditions to detect rabies virus antigen in frozen and glycerol-preserved field brain samples from northwestern Tanzania. Compared to the direct fluorescent antibody test, the traditional standard in rabies diagnosis, the dRIT was 100% sensitive and specific. PMID:16494761

  1. Canine aural cholesteatoma: a histological and immunohistochemical study.

    PubMed

    Banco, Barbara; Grieco, Valeria; Di Giancamillo, Mauro; Greci, Valentina; Travetti, Olga; Martino, Pieranna; Mortellaro, Carlo M; Giudice, Chiara

    2014-06-01

    Canine aural cholesteatoma is an epidermoid cyst that forms in the middle ear cavity as a rare complication of otitis media but the aetiopathogenesis remains controversial. In the present study, 13 cases of canine aural cholesteatoma were investigated histologically and immunohistochemically and compared with cases of chronic otitis. The immunohistochemical investigation was performed using the following monoclonal antibodies: anti-cytokeratins (CK) 14, 16, 8/18, and 19, and anti-Ki67. The proliferative indexes (PIs) of cholesteatomata and otitis epithelium were calculated as the percentage of Ki67 positive nuclei/total nuclei. Histologically, the cholesteatomata were composed of a hyperplastic, hyperkeratotic epithelium (matrix) resting on a fibrous perimatrix, infiltrated by inflammatory cells and devoid of cutaneous adnexa. Immunohistochemically, the cholesteatoma epithelium was CK14- and CK16-positive, and CK8/18- and CK19-negative. A similar pattern of CK expression was found in otitis externa. In otitis media, ciliated epithelium stained CK8/18- and CK19-positive in all layers, CK14-positive in the basal layers, and CK16-negative. The mean PIs in cholesteatomata and otitides were 18.8 and 17.8, respectively. The immunohistochemical pattern of CK expression in cholesteatomata, when compared with chronic otitis, was suggestive of hyperproliferative epithelium, but its origin could not be demonstrated. Comparable PI values were obtained in cholesteatoma and in chronic otitis, which confirmed that Ki67 is a valuable indicator of a hyperproliferative state, but not a predictor of aggressiveness. PMID:24775276

  2. Immunohistochemical characterization of 13 canine renal cell carcinomas.

    PubMed

    Gil da Costa, R M; Oliveira, J P; Saraiva, A L; Seixas, F; Faria, F; Gärtner, F; Pires, M A; Lopes, C

    2011-03-01

    Canine renal cell carcinomas (RCCs) are uncommon aggressive tumors that occur mainly in middle-aged male dogs. Their histologic classification bears no relationship with prognosis, and little information is available concerning their immunohistochemical properties. In this retrospective study, formalin-fixed, paraffin-embedded tissues from 13 canine RCCs were retrieved from the archives, classified histologically, and evaluated immunohistochemically. The dogs were 7 males and 6 females (1 spayed) of 10 different breeds, averaging 8 years in age. The tumors were classified as papillary, tubulopapillary, papillary-cystic, solid, or sarcomatoid. All 13 tumors were immunohistochemically positive for uromodulin, 12 for c-KIT, 11 for vimentin, 9 for wide-spectrum-screening cytokeratins, 7 for cytokeratins AE1/AE3 and carcinoembryonic antigen, 4 for cytokeratins CAM 5.2, and 3 for CD10. All 3 solid RCCs expressed vimentin, c-KIT, and carcinoembryonic antigen and were negative for cytokeratins. All 7 papillary and tubulopapillary tumors expressed vimentin; 6 (86%), cytokeratins; and 6 (86%), c-KIT. Both papillary-cystic RCCs were positive for cytokeratins and c-KIT and negative for vimentin. These results indicate that the different histologic types of RCC have characteristic immunohistochemical profiles and that c-KIT may be involved in the pathogenesis of canine RCC.

  3. Electrodermal activity analysis during affective haptic elicitation.

    PubMed

    Greco, Alberto; Valenza, Gaetano; Nardelli, Mimma; Bianchi, Matteo; Lanata, Antonio; Scilingo, Enzo Pasquale

    2015-08-01

    This paper investigates how the autonomic nervous system dynamics, quantified through the analysis of the electrodermal activity (EDA), is modulated according to affective haptic stimuli. Specifically, a haptic display able to convey caress-like stimuli is presented to 32 healthy subjects (16 female). Each stimulus is changed according to six combinations of three velocities and two forces levels of two motors stretching a strip of fabric. Subjects were also asked to score each stimulus in terms of arousal (high/low activation) and valence (pleasant/unpleasant), in agreement with the circumplex model of affect. EDA was processed using a deconvolutive method, separating tonic and phasic components. A statistical analysis was performed in order to identify significant differences in EDA features among force and velocity levels, as well as in their valence and arousal scores. Results show that the simulated caress induced by the haptic display significantly affects the EDA. In detail, the phasic component seems to be inversely related to the valence score. This finding is new and promising, since it can be used, e.g., as an additional cue for haptics design. PMID:26737605

  4. Analysis of drug metabolism activities in a miniaturized liver cell bioreactor for use in pharmacological studies.

    PubMed

    Hoffmann, Stefan A; Müller-Vieira, Ursula; Biemel, Klaus; Knobeloch, Daniel; Heydel, Sandra; Lübberstedt, Marc; Nüssler, Andreas K; Andersson, Tommy B; Gerlach, Jörg C; Zeilinger, Katrin

    2012-12-01

    Based on a hollow fiber perfusion technology with internal oxygenation, a miniaturized bioreactor with a volume of 0.5 mL for in vitro studies was recently developed. Here, the suitability of this novel culture system for pharmacological studies was investigated, focusing on the model drug diclofenac. Primary human liver cells were cultivated in bioreactors and in conventional monolayer cultures in parallel over 10 days. From day 3 on, diclofenac was continuously applied at a therapeutic concentration (6.4 µM) for analysis of its metabolism. In addition, the activity and gene expression of the cytochrome P450 (CYP) isoforms CYP1A2, CYP2B6, CYP2C9, CYP2D6, and CYP3A4 were assessed. Diclofenac was metabolized in bioreactor cultures with an initial conversion rate of 230 ± 57 pmol/h/10(6) cells followed by a period of stable conversion of about 100 pmol/h/10(6) cells. All CYP activities tested were maintained until day 10 of bioreactor culture. The expression of corresponding mRNAs correlated well with the degree of preservation. Immunohistochemical characterization showed the formation of neo-tissue with expression of CYP2C9 and CYP3A4 and the drug transporters breast cancer resistance protein (BCRP) and multidrug resistance protein 2 (MRP2) in the bioreactor. In contrast, monolayer cultures showed a rapid decline of diclofenac conversion and cells had largely lost activity and mRNA expression of the assessed CYP isoforms at the end of the culture period. In conclusion, diclofenac metabolism, CYP activities and gene expression levels were considerably more stable in bioreactor cultures, making the novel bioreactor a useful tool for pharmacological or toxicological investigations requiring a highly physiological in vitro representation of the liver.

  5. Immunohistochemical and molecular profiling of histologically defined apocrine carcinomas of the breast.

    PubMed

    Vranic, Semir; Marchiò, Caterina; Castellano, Isabella; Botta, Cristina; Scalzo, Maria Stella; Bender, Ryan P; Payan-Gomez, Cesar; di Cantogno, Ludovica Verdun; Gugliotta, Patrizia; Tondat, Fabrizio; di Celle, Paola Francia; Mariani, Sara; Gatalica, Zoran; Sapino, Anna

    2015-09-01

    Despite the marked improvement in the understanding of molecular mechanisms and classification of apocrine carcinoma, little is known about its specific molecular genetic alterations and potentially targetable biomarkers. In this study, we explored immunohistochemical and molecular genetic characteristics of 37 invasive apocrine carcinomas using immunohistochemistry (IHC), fluorescent in situ hybridization (FISH), multiplex ligation-dependent probe amplification (MLPA), and next-generation sequencing (NGS) assays. IHC revealed frequent E-cadherin expression (89%), moderate (16%) proliferation activity [Ki-67, phosphohistone H3], infrequent (~10%) expression of basal cell markers [CK5/6, CK14, p63, caveolin-1], loss of PTEN (83%), and overexpression of HER2 (32%), EGFR (41%), cyclin D1 (50%), and MUC-1 (88%). MLPA assay revealed gene copy gains of MYC, CCND1, ZNF703, CDH1, and TRAF4 in 50% or greater of the apocrine carcinomas, whereas gene copy losses frequently affected BRCA2 (75%), ADAM9 (54%), and BRCA1 (46%). HER2 gain, detected by MLPA in 38% of the cases, was in excellent concordance with HER2 results obtained by IHC/FISH (κ = 0.915, P < .001). TOP2A gain was observed in one case, while five cases (21%) exhibited TOP2A loss. Unsupervised hierarchical cluster analysis revealed two distinct clusters: HER2-positive and HER2-negative (P = .03 and .04, respectively). NGS assay revealed mutations of the TP53 (2 of 7, 29%), BRAF/KRAS (2 of 7, 29%), and PI3KCA/PTEN genes (7 of 7, 100%). We conclude that morphologically defined apocrine carcinomas exhibit complex molecular genetic alterations that are consistent with the "luminal-complex" phenotype. Some of the identified molecular targets are promising biomarkers; however, functional studies are needed to prove these observations.

  6. Immunohistochemical localization and quantification of the 3-(cystein-S-yl)-acetaminophen protein adduct in acetaminophen hepatotoxicity.

    PubMed Central

    Roberts, D. W.; Bucci, T. J.; Benson, R. W.; Warbritton, A. R.; McRae, T. A.; Pumford, N. R.; Hinson, J. A.

    1991-01-01

    Acetaminophen overdose causes severe hepatotoxicity in humans and laboratory animals, presumably by metabolism to N-acetyl-p-benzoquinone imine: and binding to cysteine groups as 3-(cystein-S-yl)acetaminophen-protein adduct. Antiserum specific for the adduct was used immunohistochemically to demonstrate the formation, distribution, and concentration of this specific adduct in livers of treated mice and was correlated with cell injury as a function of dose and time. Within the liver lobule, immunohistochemically demonstrable adduct occurred in a temporally progressive, central-to-peripheral pattern. There was concordance between immunohistochemical staining and quantification of the adduct in hepatic 10,000g supernate, using a quantitative particle concentration fluorescence immunoassay. Findings include: 1) immunochemically detectable adduct before the appearance of centrilobular necrosis, 2) distinctive lobular zones of adduct localization with subsequent depletion during the progression of toxicity, 3) drug-protein binding in hepatocytes at subhepatotoxic doses and before depletion of total hepatic glutathione, 4) immunohistochemical evidence of drug binding in the nucleus, and 5) adduct in metabolically active and dividing hepatocytes and in macrophagelike cells in the regenerating liver. Images Figure 2 Figure 4 PMID:1992763

  7. Intranasal Location and Immunohistochemical Characterization of the Equine Olfactory Epithelium

    PubMed Central

    Kupke, Alexandra; Wenisch, Sabine; Failing, Klaus; Herden, Christiane

    2016-01-01

    The olfactory epithelium (OE) is the only body site where neurons contact directly the environment and are therefore exposed to a broad variation of substances and insults. It can serve as portal of entry for neurotropic viruses which spread via the olfactory pathway to the central nervous system. For horses, it has been proposed and concluded mainly from rodent studies that different viruses, e.g., Borna disease virus, equine herpesvirus 1 (EHV-1), hendra virus, influenza virus, rabies virus, vesicular stomatitis virus can use this route. However, little is yet known about cytoarchitecture, protein expression and the intranasal location of the equine OE. Revealing differences in cytoarchitecture or protein expression pattern in comparison to rodents, canines, or humans might help to explain varying susceptibility to certain intranasal virus infections. On the other hand, disclosing similarities especially between rodents and other species, e.g., horses would help to underscore transferability of rodent models. Analysis of the complete noses of five adult horses revealed that in the equine OE two epithelial subtypes with distinct marker expression exist, designated as types a and b which resemble those previously described in dogs. Detailed statistical analysis was carried out to confirm the results obtained on the descriptive level. The equine OE was predominantly located in caudodorsal areas of the nasal turbinates with a significant decline in rostroventral direction, especially for type a. Immunohistochemically, olfactory marker protein and doublecortin (DCX) expression was found in more cells of OE type a, whereas expression of proliferating cell nuclear antigen and tropomyosin receptor kinase A was present in more cells of type b. Accordingly, type a resembles the mature epithelium, in contrast to the more juvenile type b. Protein expression profile was comparable to canine and rodent OE but equine types a and b were located differently within the nose and

  8. Analysis of physical activities in Taekwondo Pumsae.

    PubMed

    Lee, Sang-Bock; Cha, Eun-Jong; Lee, Tae-Soo

    2008-01-01

    Exercise is very important element for successful aging. Among many sports events, Korea is the suzerain of Taekwondo. When competing (Taekwondo Free Fighting) after learning Poomse as basic movements and inuring them, people compete with movements depending on situation. Among Poomses of Taekwondo, Taegeuk Poomse consists of very basic movements from 1 Jang to 8 Jang and they are for inuring to body. In order to prescribe Taegeuk Jang, which is the basic movement of Taekwondo that Korea is the suzerain, as an exercise for successful aging, it is necessary to analyze physical activity level of each Taegeuk Jang (From 1 Jang through 8 Jang) and suggest the same. Therefore, in this study, I analyzed physical activity level of each Jang of Taegeuk Poomse by attaching Armband made by Body Media Company on brachia and legs below knee of Taekwondo trainees. The result of the analysis of the whole momentum from Taegeuk 1 Jang to 8 Jang is as follows: First, the average absolute value of acceleration variation of vertical direction signal (L-MAD): 5.15. Second, the average absolute value of acceleration variation of horizontal direction signal (T-MAD): 3.44. Finally, the average of calorie consumption per minute (AEE/Min): 5.06 Cal. The obtained result corresponds to proper exercise condition for successful aging and it can be utilized as data for exercise prescription for the young and the old.

  9. Active polarimeter optical system laser hazard analysis.

    SciTech Connect

    Augustoni, Arnold L.

    2005-07-01

    A laser hazard analysis was performed for the SNL Active Polarimeter Optical System based on the ANSI Standard Z136.1-2000, American National Standard for Safe Use of Lasers and the ANSI Standard Z136.6-2000, American National Standard for Safe Use of Lasers Outdoors. The Active Polarimeter Optical System (APOS) uses a pulsed, near-infrared, chromium doped lithium strontium aluminum fluoride (Cr:LiSAF) crystal laser in conjunction with a holographic diffuser and lens to illuminate a scene of interest. The APOS is intended for outdoor operations. The system is mounted on a height adjustable platform (6 feet to 40 feet) and sits atop a tripod that points the beam downward. The beam can be pointed from nadir to as much as 60 degrees off of nadir producing an illuminating spot geometry that can vary from circular (at nadir) to elliptical in shape (off of nadir). The JP Innovations crystal Cr:LiSAF laser parameters are presented in section II. The illuminating laser spot size is variable and can be adjusted by adjusting the separation distance between the lens and the holographic diffuser. The system is adjusted while platform is at the lowest level. The laser spot is adjusted for a particular spot size at a particular distance (elevation) from the laser by adjusting the separation distance (d{sub diffuser}) to predetermined values. The downward pointing angle is also adjusted before the platform is raised to the selected operation elevation.

  10. Immunohistochemical study of apical periodontal cysts.

    PubMed

    Cury, V C; Sette, P S; da Silva, J V; de Araújo, V C; Gomez, R S

    1998-01-01

    Periapical lesions, as well as periodontal disease, seems to have cyclic patterns of evolution. Periods of burst may be intercalated with periods of quiescence. Because keratinocyte division must occur during cyst growth, it is presumed that epithelium status could indicate the biological activity of radicular cysts (RCs). Th1 and Th2 are important heterogeneous stages of lymphocyte differentiation. Delayed-type hypersensitivity immune reactions are mediated by Th1 lymphocytes, whereas Th2 populations mediate some types of humoral immune response. Th2 lymphocytes are characterized by high expression of CD30 glycoprotein. Th1 and Th2 lymphocytes have not been evaluated in periapical inflammatory lesions. The purpose of this study was to evaluate CD30+ cells in RCs with atrophic and hyperplastic epithelium. A biotin-streptavidin amplified system was used for identification of CD30 receptor. Results demonstrate increased proportions of Th2 cells in cysts with hyperplastic epithelium. Our results suggest that Th2 cells could be associated with RC growth.

  11. Analysis of retrotransposon activity in plants.

    PubMed

    Defraia, Christopher; Slotkin, R Keith

    2014-01-01

    Retrotransposons are transposable elements that duplicate themselves by converting their transcribed RNA genome into cDNA, which is then integrated back into the genome. Retrotransposons can be divided into two major classes based on their mechanism of transposition and the presence or absence of long terminal repeats (LTRs). In contrast to mammalian genomes, in which non-LTR retrotransposons have proliferated, plant genomes show evolutionary evidence of an explosion in LTR retrotransposon copy number. These retrotransposons can comprise a large fraction of the genome (75 % in maize). Although often viewed as molecular parasites, retrotransposons have been shown to influence neighboring gene expression and play a structural and potential regulatory role in the centromere. To prevent retrotransposon activity, eukaryotic cells have evolved overlapping mechanisms to repress transposition. Plants are an excellent system for studying the mechanisms of LTR retrotransposon inhibition such as DNA methylation and small RNA-mediated degradation of retrotransposon transcripts. However, analysis of these multi-copy, mobile elements is considerably more difficult than analysis of single-copy genes located in stable regions of the genome. In this chapter we outline methods for analyzing the progress of LTR retrotransposons through their replication cycle in plants. We describe a mixture of traditional molecular biology experiments, such as Southern, Northern, and Western blotting, in addition to nontraditional techniques designed to take advantage of the specific mechanism of LTR retrotransposition.

  12. Molecular and Immunohistochemical Characterization of Historical Long-Term Preserved Fixed Tissues from Different Human Organs

    PubMed Central

    Hühns, Maja; Röpenack, Paula; Erbersdobler, Andreas

    2015-01-01

    University and museum collections are very important sources of biological samples that can be used to asses the past and present genetic diversity of many species. Modern genetic and immunohistochemical techniques can be used on long-term preserved fixed tissues from museum specimens to answer epidemiological questions. A proof of principle was established to apply modern molecular genetics and immunohistochemical methods to these old specimens and to verify the original diagnosis. We analysed 19 specimens from our university collection including human organs that had been in fixative for more than 80 years. The tissues originated from lung, colon, brain, heart, adrenal gland, uterus and skin. We isolated amplifiable DNA from these wet preparations and performed mutational analysis of BRAF, KRAS and EGFR. The tissues were also embedded in paraffin and used for modern histology and immunohistochemistry. Our data show that amplifiable DNA is extractable and ranged from 0.25 to 22.77 μg of total DNA. In three specimens BRAFV600E or KRASG12D mutations were found. Additionally, expression of different proteins like vimentin and GFAP was detected immunohistochemical in six investigated specimens. On the basis of our results the original diagnosis was altered in three specimens. Our work showed that it is possible to extract amplifiable DNA suitable for sequence analysis from long-term fixed tissue. Furthermore, histology and immunohistochemistry is feasible in specimens fixed long time ago. We conclude that these old preparations are suitable for further epidemiological research and that our methods open up new opportunities for future studies. PMID:26252375

  13. Molecular and Immunohistochemical Characterization of Historical Long-Term Preserved Fixed Tissues from Different Human Organs.

    PubMed

    Hühns, Maja; Röpenack, Paula; Erbersdobler, Andreas

    2015-01-01

    University and museum collections are very important sources of biological samples that can be used to asses the past and present genetic diversity of many species. Modern genetic and immunohistochemical techniques can be used on long-term preserved fixed tissues from museum specimens to answer epidemiological questions. A proof of principle was established to apply modern molecular genetics and immunohistochemical methods to these old specimens and to verify the original diagnosis. We analysed 19 specimens from our university collection including human organs that had been in fixative for more than 80 years. The tissues originated from lung, colon, brain, heart, adrenal gland, uterus and skin. We isolated amplifiable DNA from these wet preparations and performed mutational analysis of BRAF, KRAS and EGFR. The tissues were also embedded in paraffin and used for modern histology and immunohistochemistry. Our data show that amplifiable DNA is extractable and ranged from 0.25 to 22.77 μg of total DNA. In three specimens BRAFV600E or KRASG12D mutations were found. Additionally, expression of different proteins like vimentin and GFAP was detected immunohistochemical in six investigated specimens. On the basis of our results the original diagnosis was altered in three specimens. Our work showed that it is possible to extract amplifiable DNA suitable for sequence analysis from long-term fixed tissue. Furthermore, histology and immunohistochemistry is feasible in specimens fixed long time ago. We conclude that these old preparations are suitable for further epidemiological research and that our methods open up new opportunities for future studies. PMID:26252375

  14. Ultra-small particles of iron oxide as peroxidase for immunohistochemical detection

    NASA Astrophysics Data System (ADS)

    Wu, Yihang; Song, Mengjie; Xin, Zhuang; Zhang, Xiaoqing; Zhang, Yu; Wang, Chunyu; Li, Suyi; Gu, Ning

    2011-06-01

    Dimercaptosuccinic acid (DMSA) modified ultra-small particles of iron oxide (USPIO) were synthesized through a two-step process. The first step: oleic acid (OA) capped Fe3O4 (OA-USPIO) were synthesized by a novel oxidation coprecipitation method in H2O/DMSO mixing system, where DMSO acts as an oxidant simultaneously. The second step: OA was replaced by DMSA to obtain water-soluble nanoparticles. The as-synthesized nanoparticles were characterized by TEM, FTIR, TGA, VSM, DLS, EDS and UV-vis. Hydrodynamic sizes and Peroxidase-like catalytic activity of the nanoparticles were investigated. The hydrodynamic sizes of the nanoparticles (around 24.4 nm) were well suited to developing stable nanoprobes for bio-detection. The kinetic studies were performed to quantitatively evaluate the catalytic ability of the peroxidase-like nanoparticles. The calculated kinetic parameters indicated that the DMSA-USPIO possesses high catalytic activity. Based on the high activity, immunohistochemical experiments were established: using low-cost nanoparticles as the enzyme instead of expensive HRP, Nimotuzumab was conjugated onto the surface of the nanoparticles to construct a kind of ultra-small nanoprobe which was employed to detect epidermal growth factor receptor (EGFR) over-expressed on the membrane of esophageal cancer cell. The proper sizes of the probes and the result of membranous immunohistochemical staining suggest that the probes can be served as a useful diagnostic reagent for bio-detection.

  15. Assessing physical activity intensity by video analysis.

    PubMed

    Silva, P; Santiago, C; Reis, L P; Sousa, A; Mota, J; Welk, G

    2015-05-01

    Assessing physical activity (PA) is a challenging task and many different approaches have been proposed. Direct observation (DO) techniques can objectively code both the behavior and the context in which it occurred, however, they have significant limitations such as the cost and burden associated with collecting and processing data. Therefore, this study evaluated the utility of an automated video analysis system (CAM) designed to record and discriminate the intensity of PA using a subject tracking methodology. The relative utility of the CAM system and DO were compared with criterion data from an objective accelerometry-based device (Actigraph GT3X+). Eight 10 year old children (three girls and five boys) wore the GT3X+ during a standard basketball session. PA was analyzed by two observers using the SOPLAY instrument and by the CAM system. The GT3X+ and the CAM were both set up to collect data at 30 Hz while the DO was performed every two minutes, with 10 s of observation for each gender. The GT3X+ was processed using cut points by Evanson and the outcome measure was the percentage of time spent in different intensities of PA. The CAM data were processed similarly using the same speed thresholds as were used in establishing the Evenson cut-off points (light: <2 mph; walking: 2-4 mph; very active: >4 mph). Similar outcomes were computed from the SOPLAY default analyses. A chi-square test was used to test differences in the percentage of time at the three intensity zones (light, walking and very active). The Yates' correction was used to prevent overestimation of statistical significance for small data. When compared with GT3X+, the CAM had better results than the SOPLAY. The chi-square test yielded the following pairwise comparisons: CAM versus GT3x+ was χ(2) (5) = 24.18, p < .001; SOPLAY2 versus GT3x+ was χ(2) (5) = 144.44, p < .001; SOPLAY1 versus GT3x+ was χ(2) (5) = 119.55, p < .001. The differences were smaller between CAM and GT3x

  16. Solitary fibrous tumors of the pleura: clinicopathological and immunohistochemical examination.

    PubMed

    Hiraoka, Kei; Morikawa, Toshiaki; Ohbuchi, Toshiro; Katoh, Hiroyuki

    2003-03-01

    The purpose of this work was to study clinical and biological characteristics of solitary fibrous tumor (SFT) of the pleura. We reviewed the clinicopathological and immunohistochemical features of 12 patients who underwent surgical resection for SFT. Ten cases were histologically defined as benign; two were found to be malignant. CD34 negativity and strong expression of p53 could be observed in a patient with fatal outcome. Ki-67 expression was increased in malignant cases, as compared with benign. We also found that Bcl-2 expression inversely correlated with a tumor diameter. As the development of malignant SFT might be associated with these molecular statuses, immunohistochemical staining should be performed in all cases to identify the biological characteristics of the tumor.

  17. Comparison between histochemical and immunohistochemical methods for diagnosis of sporotrichosis.

    PubMed Central

    Marques, M E; Coelho, K I; Sotto, M N; Bacchi, C E

    1992-01-01

    AIMS: To compare the efficacy of histochemical and immunohistochemical methods in detecting forms of Sporothrix schenckii in tissue. METHODS: Thirty five cutaneous biopsy specimens from 27 patients with sporotrichosis were stained by histochemical haematoxylin and eosin, periodic acid Schiff, and Gomori's methenamine silver methods and an immunohistochemical (avidin-biotin complex immunoperoxidase) (ABC) technique associated with a newly produced rabbit polyclonal antibody anti-Sporothrix schenckii. RESULTS: A total of 29 (83%) cases were positive by the ABC method used in association with anti-Sporothrix schenckii rabbit polyclonal antibodies. Histochemical methods, using silver staining, periodic acid Schiff, and conventional haematoxylin and eosin detected 37%, 23%, and 23% of forms of S schenckii, respectively. The ABC technique was significantly more reliable than periodic acid Schiff and silver staining techniques. CONCLUSIONS: It is concluded that immunostaining is an easy and rapid method which can efficiently increase the accuracy of the diagnosis of sporotrichosis in human tissue. Images PMID:1479036

  18. Immunohistochemical localization of carcinoembryonic antigen (CEA) in salivary gland tumors.

    PubMed

    Alfaro, M; Carrozza, M

    1990-04-01

    Carcinoembryonic antigen (CEA) was first isolated from colonic carcinoma and has been used as a diagnostic marker. CEA has also been observed in a variety of epithelial tumors and normal tissues. In this study, CEA was localized by means of immunohistochemical procedures in benign and malignant salivary gland tumors, as well as in normal parotid gland, indicating that CEA is not a reliable marker for differentiation between benign and malignant salivary gland neoplasms.

  19. Immunohistochemical investigation of the coma blister and its pathogenesis.

    PubMed

    Kashiwagi, Masayuki; Ishigami, Akiko; Hara, Kenji; Matsusue, Aya; Waters, Brian; Takayama, Mio; Tokunaga, Itsuo; Nishimura, Akiyoshi; Kubo, Shin-ichi

    2013-01-01

    The erythematous patches and vesicles that are observed in coma patients, usually from an overdose of medication, are known as coma blisters. However, it is unknown whether the degenerated sweat gland is a necrosis or apoptosis. We immunohistochemically examined such skin lesions to investigate the characteristics and pathogenesis of the coma blister. Skin lesions were obtained from a forensic autopsy case, a woman in her thirties, of caffeine intoxication. Those lesions were observed in the left femoral, the lower left thigh, and the right knee. Histologically, the skin lesions showed that the keratinocytes had necrosed and the epidermis was thin in some areas. Eccrine sweat gland degeneration was observed. Obvious inflammatory cell infiltrations were not detected. Immunohistochemically, we stained each skin lesion against CD3, CD8, CD45RO, cytokeratin, 70 kD heat shock protein, ubiquitin, 150 kD oxygen regulated protein, and caspase-cleaved keratin 18 neo-epitope M30. They were also stained with an in situ apoptosis detection kit. Degenerated sweat glands featured CD45RO and M30 immunoreactivity. Immunohistochemical staining for CD45RO, CK-L, and M30 might be useful to observe sweat gland degeneration in the coma blister. Therefore, the apoptosis might be related to coma blisters and sweat gland degenerations.

  20. Prostatic tissue in testicular teratoma. A clinicopathologic and immunohistochemical study.

    PubMed

    Roma, Andres A; Humphrey, Peter A

    2013-02-01

    The presence of prostatic differentiation as part of teratoma is very unusual and has been reported less than 20 times in the literature; however, all but 1 case were described in ovarian teratomas. We reviewed 45 specimens of germ cell tumors with teratoma component in postpuberal male patients. Original hematoxylin and eosin review failed to identify glands morphologically consistent with prostatic differentiation. Immunohistochemical stains performed on 10 specimens from 10 patients with small glandular and/or tubular structures revealed 1 case with glands positive for prostatic-specific antigen, prostatic-specific acid phosphatase, and prostein/P501S, whereas high-molecular-weight cytokeratin and p63 highlighted only basal cells. The glands were irregular in size and shape and contained mostly cuboidal to columnar luminal-type cells with occasional basal-type cells. Re-review of all the specimens revealed a second block from the same testis as well as 1 retroperitoneal lymph node with metastatic teratoma in the same patient, also immunohistochemically confirmed. These glands were seen in a smooth muscle stromal background, adjacent to classic gastrointestinal and tracheobronchial teratoma components. Our findings show immunohistochemically confirmed prostatic differentiation in 2 specimens from 1 patient with teratoma. This study raises the possibility that prostatic differentiation, difficult to recognize on morphology alone, might not be that unusual and that immunostains can help detect it over the several different epithelial components of teratoma.

  1. Immunohistochemical phenotype and molecular pathological characteristics of metanephric adenoma.

    PubMed

    Sun, Zhulei; Kan, Shihai; Zhang, Leilei; Zhang, Yan; Jing, Hong; Huang, Gui; Yu, Qichun; Wu, Jiang

    2015-01-01

    To assess the clinicopathological, immunohistochemical and molecular features of metanephric adenoma (MA). Clinicopathologic data were obtained for 5 cases of MA with follow-up information. Specimens from these patients were stained by HE and immunohistochemistry for the detection of WT1, vimentin, S-100 protein, CK7, P504s, CD10 and renal cell carcinoma marker (RCC). Fluorescence in situ hybridization (FISH) was performed on 4 tumors. The patients included 1 male and 4 females, aged from 30 to 49 (mean=39) years. Tumor diameters ranged from 3 to 5.5 cm. Histologically, the tumors had tubular, papillary, or glomeruloid architectures, and were composed of cells with uniform and round nuclei, inconspicuous nucleoli, and high ratio of nucleus to cytoplasm. Nuclear polymorphism and mitotic figures were not observed. Immunohistochemically, they expressed WT1 (5/5), vimentin (5/5), S-100 (4/5), CK7 (2/5), P504s (2/5), and CD10 (1/5) and not RCC. FISH study was carried out on 4 metanephric adenoma cases, and no abnormalities were observed in chromosomes 3, 7, 17, and P16 gene of chromosomes 9. MA is an uncommon renal tumor. Its diagnosis depends on morphological, immunohistochemical and molecular features. PMID:26261480

  2. Immunohistochemical Analysis of Matrix Metalloproteinase-13 in Human Caries Dentin

    PubMed Central

    Loreto, C.; Galanti, C.; Musumeci, G.; Rusu, M.C.; Leonardi, R.

    2014-01-01

    The immunoexpression profile of matrix metalloproteinase-13 was investigated for the first time in dentin of human caries and healthy teeth. Twelve permanent premolars (10 caries and 2 sound) were decalcified in ethylenediaminetetraacetic acid and processed for embedding in paraffin wax. Sections 3-4 µm in thickness were cut and processed for immunohistochemistry. A mouse monoclonal anti-metalloproteinase-13 antibody was used for localisation using an immunoperoxidase technique. Dentinal immunoreactivity was detected in all teeth; it was weak in sound teeth and strong close to the caries area. These in vivo findings suggest a role for metalloproteinase-13 in the development and progression of adult human dental tissue disorders. PMID:24704999

  3. Malignant dermatofibroma: clinicopathological, immunohistochemical, and molecular analysis of seven cases.

    PubMed

    Mentzel, Thomas; Wiesner, Thomas; Cerroni, Lorenzo; Hantschke, Markus; Kutzner, Heinz; Rütten, Arno; Häberle, Michael; Bisceglia, Michele; Chibon, Frederic; Coindre, Jean-Michel

    2013-02-01

    Dermatofibroma (cutaneous fibrous histiocytoma) represents a common benign mesenchymal tumor, and numerous morphological variants have been described. Some variants of dermatofibroma are characterized by an increased risk of local recurrences, and there are a few reported metastasizing cases. Unfortunately, an aggressive behavior cannot be predicted reliably by morphology at the moment, and we evaluated the value of array-comparative genomic hybridization (CGH) in this setting. Seven cases of clinically aggressive dermatofibromas were identified, and pathological and molecular features were evaluated. The neoplasms occurred in four female and in three male patients (mean age was 33 years, range 2-65 years), and arose on the shoulder, buttock, temple, lateral neck, thigh, ankle, and cheek. The size of the neoplasms ranged from 1 to 9 cm (mean: 3 cm). An infiltration of the subcutis was seen in five cases. Two neoplasms were completely excised, whereas an incomplete or marginal excision was reported in the remaining cases. Local recurrences were seen in six cases (time to the first recurrence ranged from 8 months to 9 years). Metastases were noted between 3 months and 8 years after diagnosis in six patients. Two patients died of disease, and two patients are alive with disease. Histologically, the primary tumors showed features of cellular dermatofibroma (four cases), cellular/aneurysmal dermatofibroma (one case), atypical/cellular dermatofibroma (one case), and classical dermatofibroma (one case). Mitotic figures ranged from 3 to 25 per 10 high-power fields, and focal necrosis was present in five cases. Interestingly, malignant transformation from cellular dermatofibroma to an obvious spindle cell/pleomorphic sarcoma was seen in one primary and in one recurrent neoplasm. Five neoplasms showed chromosomal aberrations by array-CGH, suggesting that these changes may represent an additional diagnostic tool in the recognition of cases of dermatofibroma with a metastatic potential. PMID:22996372

  4. Activation Likelihood Estimation meta-analysis revisited

    PubMed Central

    Eickhoff, Simon B.; Bzdok, Danilo; Laird, Angela R.; Kurth, Florian; Fox, Peter T.

    2011-01-01

    A widely used technique for coordinate-based meta-analysis of neuroimaging data is activation likelihood estimation (ALE), which determines the convergence of foci reported from different experiments. ALE analysis involves modelling these foci as probability distributions whose width is based on empirical estimates of the spatial uncertainty due to the between-subject and between-template variability of neuroimaging data. ALE results are assessed against a null-distribution of random spatial association between experiments, resulting in random-effects inference. In the present revision of this algorithm, we address two remaining drawbacks of the previous algorithm. First, the assessment of spatial association between experiments was based on a highly time-consuming permutation test, which nevertheless entailed the danger of underestimating the right tail of the null-distribution. In this report, we outline how this previous approach may be replaced by a faster and more precise analytical method. Second, the previously applied correction procedure, i.e. controlling the false discovery rate (FDR), is supplemented by new approaches for correcting the family-wise error rate and the cluster-level significance. The different alternatives for drawing inference on meta-analytic results are evaluated on an exemplary dataset on face perception as well as discussed with respect to their methodological limitations and advantages. In summary, we thus replaced the previous permutation algorithm with a faster and more rigorous analytical solution for the null-distribution and comprehensively address the issue of multiple-comparison corrections. The proposed revision of the ALE-algorithm should provide an improved tool for conducting coordinate-based meta-analyses on functional imaging data. PMID:21963913

  5. Enzyme catalysis enhanced dark-field imaging as a novel immunohistochemical method

    NASA Astrophysics Data System (ADS)

    Fan, Lin; Tian, Yanyan; Yin, Rong; Lou, Doudou; Zhang, Xizhi; Wang, Meng; Ma, Ming; Luo, Shouhua; Li, Suyi; Gu, Ning; Zhang, Yu

    2016-04-01

    Conventional immunohistochemistry is limited to subjective judgment based on human experience and thus it is clinically required to develop a quantitative immunohistochemical detection. 3,3'-Diaminobenzidin (DAB) aggregates, a type of staining product formed by conventional immunohistochemistry, were found to have a special optical property of dark-field imaging for the first time, and the mechanism was explored. On this basis, a novel immunohistochemical method based on dark-field imaging for detecting HER2 overexpressed in breast cancer was established, and the quantitative analysis standard and relevant software for measuring the scattering intensity was developed. In order to achieve a more sensitive detection, the HRP (horseradish peroxidase)-labeled secondary antibodies conjugated gold nanoparticles were constructed as nanoprobes to load more HRP enzymes, resulting in an enhanced DAB deposition as a dark-field label. Simultaneously, gold nanoparticles also act as a synergistically enhanced agent due to their mimicry of enzyme catalysis and dark-field scattering properties.Conventional immunohistochemistry is limited to subjective judgment based on human experience and thus it is clinically required to develop a quantitative immunohistochemical detection. 3,3'-Diaminobenzidin (DAB) aggregates, a type of staining product formed by conventional immunohistochemistry, were found to have a special optical property of dark-field imaging for the first time, and the mechanism was explored. On this basis, a novel immunohistochemical method based on dark-field imaging for detecting HER2 overexpressed in breast cancer was established, and the quantitative analysis standard and relevant software for measuring the scattering intensity was developed. In order to achieve a more sensitive detection, the HRP (horseradish peroxidase)-labeled secondary antibodies conjugated gold nanoparticles were constructed as nanoprobes to load more HRP enzymes, resulting in an enhanced DAB

  6. Development of an immunohistochemical protein quantification system in conjunction with tissue microarray technology for identifying predictive biomarkers for phosphatidylinositol 3-kinase inhibitors.

    PubMed

    Isoyama, Sho; Yoshimi, Hisashi; Dan, Shingo; Okamura, Mutsumi; Seki, Mariko; Irimura, Tatsuro; Yamori, Takao

    2012-01-01

    The phosphatidylinositol 3-kinase (PI3K) pathway is frequently activated in human cancers by gain-of-function mutations of phosphoinositide-3-kinase, catalytic, alpha polypeptide (PIK3CA) or dysfunction of phosphatase and tensin homolog deleted on chromosome 10 (PTEN). Therefore PI3K is thought to be a promising target for cancer therapy. Many agents targeting PI3K have been developed and some of them have been evaluated in clinical trials. In recent years, development of predictive biomarkers as companion diagnostics for molecular targeted drugs has become an important requirement for clinical development; however, no clinically established biomarkers that predict the efficacy of PI3K inhibitors have been found. We previously reported that expression of phosphorylated Akt determined by immunoblot analysis correlated with the antitumor efficacy of a PI3K inhibitor ZSTK474 in vitro and in vivo, suggesting that it might be used as a predictive biomarker. In this study, to evaluate biomarker candidates in in vivo tumor samples, we developed an immunohistochemical protein detection/quantification system in conjunction with the tissue microarray technology using a panel of 24 human tumor xenografts (JFCR24). We have clearly demonstrated that expression levels of phosphorylated v-akt murine thymoma viral oncogene homolog (Akt) and mitogen-activated protein kinase (MAPK) determined by this system significantly correlated with those determined by immunoblot analysis. As expected, PTEN status correlated with expression of phosphorylated Akt but not MAPK. Finally, we confirmed that phosphorylated Akt levels determined using this system correlated with the in vivo efficacy of ZSTK474. The present results indicate that the immunohistochemical protein detection/quantification system could be used to quantify expression of biomarker proteins in xenografted tumor tissues as well as in human tumor specimens to predict drug efficacy in future clinical trials. PMID:22975517

  7. Cystic Renal Oncocytoma and Tubulocystic Renal Cell Carcinoma: Morphologic and Immunohistochemical Comparative Study.

    PubMed

    Skenderi, Faruk; Ulamec, Monika; Vranic, Semir; Bilalovic, Nurija; Peckova, Kvetoslava; Rotterova, Pavla; Kokoskova, Bohuslava; Trpkov, Kiril; Vesela, Pavla; Hora, Milan; Kalusova, Kristyna; Sperga, Maris; Perez Montiel, Delia; Alvarado Cabrero, Isabel; Bulimbasic, Stela; Branzovsky, Jindrich; Michal, Michal; Hes, Ondrej

    2016-02-01

    Renal oncocytoma (RO) may present with a tubulocystic growth in 3% to 7% of cases, and in such cases its morphology may significantly overlap with tubulocystic renal cell carcinoma (TCRCC). We compared the morphologic and immunohistochemical characteristics of these tumors, aiming to clarify the differential diagnostic criteria, which facilitate the discrimination of RO from TCRCC. Twenty-four cystic ROs and 15 TCRCCs were selected and analyzed for: architectural growth patterns, stromal features, cytomorphology, ISUP nucleolar grade, necrosis, and mitotic activity. Immunohistochemical panel included various cytokeratins (AE1-AE3, OSCAR, CAM5.2, CK7), vimentin, CD10, CD117, AMACR, CA-IX, antimitochondrial antigen (MIA), EMA, and Ki-67. The presence of at least focal solid growth and islands of tumor cells interspersed with loose stroma, lower ISUP nucleolar grade, absence of necrosis, and absence of mitotic figures were strongly suggestive of a cystic RO. In contrast, the absence of solid and island growth patterns and presence of more compact, fibrous stroma, accompanied by higher ISUP nucleolar grade, focal necrosis, and mitotic figures were all associated with TCRCC. TCRCC marked more frequently for vimentin, CD10, AMACR, and CK7 and had a higher proliferative index by Ki-67 (>15%). CD117 was negative in 14/15 cases. One case was weakly CD117 reactive with cytoplasmic positivity. All cystic RO cases were strongly positive for CD117. The remaining markers (AE1-AE3, CAM5.2, OSCAR, CA-IX, MIA, EMA) were of limited utility. Presence of tumor cell islands and solid growth areas and the type of stroma may be major morphologic criteria in differentiating cystic RO from TCRCC. In difficult cases, or when a limited tissue precludes full morphologic assessment, immunohistochemical pattern of vimentin, CD10, CD117, AMACR, CK7, and Ki-67 could help in establishing the correct diagnosis. PMID:26180933

  8. Small bowel transplantation in children: an immunohistochemical study of intestinal grafts.

    PubMed Central

    Fromont, G; Cerf-Bensussan, N; Patey, N; Canioni, D; Rambaud, C; Goulet, O; Jan, D; Révillon, Y; Ricour, C; Brousse, N

    1995-01-01

    Seven children with short bowel syndrome underwent small bowel allografting. Episodes of early rejection were observed in five patients who received a graft from paediatric or adult donors but not in two patients who received a neonatal graft. This study aimed, firstly, to define immunohistochemical parameters accompanying rejection and, secondly, to compare immunohistochemical parameters in neonatal grafts with those in grafts from older donors. An immunohistochemical analysis was performed on 85 intestinal biopsy specimens taken for monitoring the transplant. Acute histological rejection was associated with pericryptic infiltrates of CD3+TcR alpha beta + T cells containing clusters of CD8+ cells, numerous CD25+ cells, and increased numbers of CD68+ macrophages. These changes were associated with the appearance of major histocompatibility (MHC) class II antigens on crypt enterocytes and with an appreciable increase in the expression of E-selectin on mucosal endothelial cells. Immunohistochemistry was useful in predicting rejection by showing the appearance of pericryptic CD25+ T cells 48 hours before the first histological lesions of crypt necrosis. Comparison of neonatal grafts with grafts from older donors did not show any significant difference in the density of CD68+ macrophages or in the endothelial expression of intercellular adhesion molecule-1, vascular cell adhesion molecule-1, or E-selectin. In contrast to grafts from older donors, however, neonatal grafts did not express MHC class II antigens on epithelial cells and contained very low numbers of intraepithelial lymphocytes. These data indicate, firstly, that immunohistochemistry is useful for monitoring intestinal transplants and, secondly, that the better clinical tolerance of neonatal allografts may be related to the lower immunogenicity of the neonatal epithelium. Images Figure 3 Figure 4 PMID:8537048

  9. Immunohistochemical prediction of lapatinib efficacy in advanced HER2-positive breast cancer patients

    PubMed Central

    Duchnowska, Renata; Wysocki, Piotr J.; Korski, Konstanty; Czartoryska-Arłukowicz, Bogumiła; Niwińska, Anna; Orlikowska, Marlena; Radecka, Barbara; Studziński, Maciej; Demlova, Regina; Ziółkowska, Barbara; Merdalska, Monika; Hajac, Łukasz; Myśliwiec, Paulina; Zuziak, Dorota; Dębska-Szmich, Sylwia; Lang, Istvan; Foszczyńska-Kłoda, Małgorzata; Karczmarek-Borowska, Bożenna; Żawrocki, Anton; Kowalczyk, Anna; Biernat, Wojciech; Jassem, Jacek

    2016-01-01

    Molecular mechanisms of lapatinib resistance in breast cancer are not well understood. The aim of this study was to correlate expression of selected proteins involved in ErbB family signaling pathways with clinical efficacy of lapatinib. Study group included 270 HER2-positive advanced breast cancer patients treated with lapatinib and capecitabine. Immunohistochemical expression of phosphorylated adenosine monophosphate-activated protein (p-AMPK), mitogen-activated protein kinase (p-MAPK), phospho (p)-p70S6K, cyclin E, phosphatase and tensin homolog were analyzed in primary breast cancer samples. The best discriminative value for progression-free survival (PFS) was established for each biomarker and subjected to multivariate analysis. At least one biomarker was determined in 199 patients. Expression of p-p70S6K was independently associated with longer (HR 0.45; 95% CI: 0.25–0.81; p = 0.009), and cyclin E with shorter PFS (HR 1.83; 95% CI: 1.06–3.14; p = 0.029). Expression of p-MAPK (HR 1.61; 95% CI 1.13–2.29; p = 0.009) and cyclin E (HR 2.99; 95% CI: 1.29–6.94; p = 0.011) was correlated with shorter, and expression of estrogen receptor (HR 0.65; 95% CI 0.43–0.98; p = 0.041) with longer overall survival. Expression of p-AMPK negatively impacted response to treatment (HR 3.31; 95% CI 1.48–7.44; p = 0.004) and disease control (HR 3.07; 95% CI 1.25–7.58; p = 0.015). In conclusion: the efficacy of lapatinib seems to be associated with the activity of downstream signaling pathways – AMPK/mTOR and Ras/Raf/MAPK. Further research is warranted to assess the clinical utility of these data and to determine a potential role of combining lapatinib with MAPK pathway inhibitors. PMID:26623720

  10. Commercial horseradish peroxidase degrades myelin encephalitogenic protein during coupling for immunohistochemical studies.

    PubMed

    Johnson, A B; Cammer, W

    1977-05-01

    Conjugates of myelin encephalitogenic basic protein (EP) and commercial horseradish peroxidase (HRP) have been used for immunohistochemical demonstrations of anti-EP antibody in animals with experimental allergic encephalomyelitis. We performed gel electrophoresis studies on EP-HRP conjugates prepared with glutaraldehyde and on mixtures of EP and HRP incubated without glutaraldehyde. The results show that under conditions of one-and two-step coupling HRP causes rapid loss of the native EP band, apparently due to EP degradation. The EP-HRP mixtures are not encephalitogenic in rabbits, or encephalitogenic activity is lost during processing. The immunohistochemical reactivity of conjugates, however, signals some preservation of antibody-combining sites. The mechanism of the HRP effect on EP is unknown. The possibilities of a contaminating proteinase or direct peroxidatic attack are suggested. Until this action of HRP can be overcome, the effect of coupling procedures on the biological activities of EP will be difficult to assess, and EP-HRP conjugates cannot be expected to reveal sites that may bind encephalitogenic portions of the EP molecule.

  11. An immunohistochemical localization of neuropeptide Y (NPY) in its amidated form in human frontal cortex.

    PubMed

    Blinkenberg, M; Kruse-Larsen, C; Mikkelsen, J D

    1990-01-01

    The distribution of neuropeptide Y (NPY)-immunoreactive neurons was studied in human frontal cerebral cortex from surgical biopsy specimens by immunohistochemical techniques. NPY-containing neurons were identified in all cortical sublayers except sublayer I. The stained neurons were of the multipolar, bitufted, round or triangular form with dendritic and axonal processes. The immunoreactive neurons were considered to be cortical interneurons, due to their nonpyramidal form, and since their processes could be followed intracortically particularly in direction to superficial cortical layers. The NPY precursor molecule is processed to NPY by a dibasic cleavage, and NPY is further enzymatically amidated before release and receptor activation can be achieved. Antisera raised against Cys-NPY(32-36)amide recognize amidated NPY not cross-reacting with nonamidated NPY. These antisera and immunohistochemistry revealed the presence of a population of NPYamide-immunoreactive cells morphologically indistinguishable from the NPY-immunoreactive cells in the human frontal cortex. By comparing the number of immunoreactive cells in adjacent sections, it appears that the number of NPY-immunoreactive cells was higher than those immunoreactive to NPYamide. Also, the density of NPY fibers was much higher compared with the number stained with NPYamide antiserum. The present immunohistochemical study indicates that NPY in its amidated form is contained in a subpopulation of human cortical NPY-immunoreactive neurons and may participate as an active neurotransmitter/modulator within the human cerebral cortex.

  12. Biochemical and immunohistochemical comparison of collagen in granuloma annulare and skin sarcoidosis.

    PubMed

    Oikarinen, A; Kinnunen, T; Kallioinen, M

    1989-01-01

    Collagen was studied by biochemical and immunohistochemical means in 5 patients with granuloma annulare (GA) and 3 with cutaneous sarcoidosis (SA). The solubility of collagen from the lesional skin in acetic acid was higher than that of collagen from unaffected skin from both patients and control subjects. Collagen concentration in the skin lesions, measured in terms of hydroxyproline content, was reduced in 3 patients with granuloma annulare and one with sarcoidosis, but the ratio of type III/I collagen was unchanged vis-à-vis non-affected skin. The collagen concentration in non-affected skin of both GA and SA-patients was also lower than in controls. The most typical immunohistochemical finding was the association of type III procollagen and fibronectin with granulomas in the lesional skin of both GA and SA cases. The activity of prolyl hydroxylase, a key enzyme in collagen biosynthesis, was markedly increased in the lesional skin, indicating that collagen synthesis in vivo was also increased. Surprisingly, collagen synthesis was not increased in cell culture studies. This could be due to cell selection as observed previously in scleroderma. Another possibility could be that various mediators released in vivo from inflammatory cells activate fibroblasts. However, when cells are subcultivated, this effect is not maintained. In conclusion, marked changes in collagen could be observed in granuloma annulare and skin sarcoidosis, reflecting increased turnover of collagen in vivo.

  13. Langerhans cells in lichen planus and lichenoid mucositis an immunohistochemical study

    PubMed Central

    Devi, M.; Saraswathi, T. R.; Ranganathan, K.; Vijayalakshmi, D.; Sreeja, C.; Fathima, S. Shabana

    2014-01-01

    Aim: The aim of this study is to identify and evaluate Langerhans cell (LC) in lichen planus (LP), lichenoid mucositis (LM) and normal mucosa (NM) using CD1a monoclonal antibody immunohistochemically. Materials and Methods: A total of 15 cases of oral lichen planus and 15 cases of LM were selected based on clinical examination and confirmed by histopathological analysis. The biopsies from the 10 patients were taken from normal buccal mucosa as control. Paraffin blocks of tissue were made, which are used for routine hematoxylin and eosin staining and immunohistochemical staining using biotin streptavidin methods (CD1a monoclonal antibody). Analysis of CD1a expression was performed by evaluating the labeling index (LI) for each slide. Results: The mean CD1a LI for LP was significantly higher than that of LM and NM in the basal and supra basal layer. The mean CD1a positive cells in the connective tissues for LP were higher than that of LM and NM. Conclusion: This study clearly demonstrates a statistically significant increase in number of LC in LP than in LM, indicating the possible different immunopathogenic mechanisms. PMID:25210358

  14. Effects of Image Compression on Automatic Count of Immunohistochemically Stained Nuclei in Digital Images

    PubMed Central

    López, Carlos; Lejeune, Marylène; Escrivà, Patricia; Bosch, Ramón; Salvadó, Maria Teresa; Pons, Lluis E.; Baucells, Jordi; Cugat, Xavier; Álvaro, Tomás; Jaén, Joaquín

    2008-01-01

    This study investigates the effects of digital image compression on automatic quantification of immunohistochemical nuclear markers. We examined 188 images with a previously validated computer-assisted analysis system. A first group was composed of 47 images captured in TIFF format, and other three contained the same images converted from TIFF to JPEG format with 3×, 23× and 46× compression. Counts of TIFF format images were compared with the other three groups. Overall, differences in the count of the images increased with the percentage of compression. Low-complexity images (≤100 cells/field, without clusters or with small-area clusters) had small differences (<5 cells/field in 95–100% of cases) and high-complexity images showed substantial differences (<35–50 cells/field in 95–100% of cases). Compression does not compromise the accuracy of immunohistochemical nuclear marker counts obtained by computer-assisted analysis systems for digital images with low complexity and could be an efficient method for storing these images. PMID:18755997

  15. Effect of hydroalcoholic extract of Anethum graveolens leaves on the dentate gyrus of the hippocampus in the epileptic mice: a histopathological and immunohistochemical study

    PubMed Central

    Golmohammadi, Rahim; Sabaghzadeh, Fatemeh; Mojadadi, Mohammad Shafi

    2016-01-01

    Anethum graveolens or Dill (local name: Shevid) belongs to the family of Apiaceae (Umbelliferae) and is used traditionally for the treatment of convulsion and diabetes in Iran. This study aimed to investigate the effect of hydroalcoholic extract of A. graveolens leaves on the histology of the dentate gyrus of the hippocampus in the epileptic mice kindled by Pentylenetetrazole (PTZ). In this experimental study, the epileptic BALB/c mice kindled by PTZ were randomly divided into four groups of 10 animals each. Three experimental groups received 250, 500 and 750 mg/kg/day of A. graveolens extract for 21 days. The control group received phosphate-buffered saline (PBS). After the treatment period, the mice were anesthetized, and their hippocampi were dissected for the histopathological analysis, and immunohistochemical analysis for caspase-3 activity. Histopathological examinations showed that the mean numbers of the healthy neuronal cells in the dentate gyrus of the mice received 500 mg/kg/day of A. graveolens extracts were significantly higher than those of the mice received 250 and 750 mg/kg/day of the extracts as well as the control group (P < 0.05 and P < 0.001, respectively). In addition, the results of immunohistochemical analysis revealed that in mice treated with 500 mg/kg/day of A. graveolens; the numbers of caspase-3-positive cells in the dentate gyrus were significantly lower than those of the two other test and the control groups. The findings of this study suggest that 500 mg/kg/day of the A. graveolens extract could have protective effect on the dentate gyrus of the hippocampus in the epileptic mice. PMID:27499792

  16. Effect of hydroalcoholic extract of Anethum graveolens leaves on the dentate gyrus of the hippocampus in the epileptic mice: a histopathological and immunohistochemical study.

    PubMed

    Golmohammadi, Rahim; Sabaghzadeh, Fatemeh; Mojadadi, Mohammad Shafi

    2016-01-01

    Anethum graveolens or Dill (local name: Shevid) belongs to the family of Apiaceae (Umbelliferae) and is used traditionally for the treatment of convulsion and diabetes in Iran. This study aimed to investigate the effect of hydroalcoholic extract of A. graveolens leaves on the histology of the dentate gyrus of the hippocampus in the epileptic mice kindled by Pentylenetetrazole (PTZ). In this experimental study, the epileptic BALB/c mice kindled by PTZ were randomly divided into four groups of 10 animals each. Three experimental groups received 250, 500 and 750 mg/kg/day of A. graveolens extract for 21 days. The control group received phosphate-buffered saline (PBS). After the treatment period, the mice were anesthetized, and their hippocampi were dissected for the histopathological analysis, and immunohistochemical analysis for caspase-3 activity. Histopathological examinations showed that the mean numbers of the healthy neuronal cells in the dentate gyrus of the mice received 500 mg/kg/day of A. graveolens extracts were significantly higher than those of the mice received 250 and 750 mg/kg/day of the extracts as well as the control group (P < 0.05 and P < 0.001, respectively). In addition, the results of immunohistochemical analysis revealed that in mice treated with 500 mg/kg/day of A. graveolens; the numbers of caspase-3-positive cells in the dentate gyrus were significantly lower than those of the two other test and the control groups. The findings of this study suggest that 500 mg/kg/day of the A. graveolens extract could have protective effect on the dentate gyrus of the hippocampus in the epileptic mice. PMID:27499792

  17. Analysis of mitogen-activated protein kinase activity in yeast.

    PubMed

    Elion, Elaine A; Sahoo, Rupam

    2010-01-01

    Mitogen-activated protein (MAP) kinases play central roles in transmitting extracellular and intracellular information in a wide variety of situations in eukaryotic cells. Their activities are perturbed in a large number of diseases, and their activating kinases are currently therapeutic targets in cancer. MAPKs are highly conserved among all eukaryotes. MAPKs were first cloned from the yeast Saccharomyces cerevisiae. Yeast has five MAPKs and one MAPK-like kinase. The mating MAPK Fus3 is the best characterized yeast MAPK. Members of all subfamilies of human MAPKs can functionally substitute S. cerevisiae MAPKs, providing systems to use genetic approaches to study the functions of either yeast or human MAPKs and to identify functionally relevant amino acid residues that enhance or reduce the effects of therapeutically relevant inhibitors and regulatory proteins. Here, we describe an assay to measure Fus3 activity in immune complexes prepared from S. cerevisiae extracts. The assay conditions are applicable to other MAPKs, as well. PMID:20811996

  18. Single Cell Analysis of Transcriptional Activation Dynamics

    PubMed Central

    Rafalska-Metcalf, Ilona U.; Powers, Sara Lawrence; Joo, Lucy M.; LeRoy, Gary; Janicki, Susan M.

    2010-01-01

    Background Gene activation is thought to occur through a series of temporally defined regulatory steps. However, this process has not been completely evaluated in single living mammalian cells. Methodology/Principal Findings To investigate the timing and coordination of gene activation events, we tracked the recruitment of GCN5 (histone acetyltransferase), RNA polymerase II, Brd2 and Brd4 (acetyl-lysine binding proteins), in relation to a VP16-transcriptional activator, to a transcription site that can be visualized in single living cells. All accumulated rapidly with the VP16 activator as did the transcribed RNA. RNA was also detected at significantly more transcription sites in cells expressing the VP16-activator compared to a p53-activator. After α-amanitin pre-treatment, the VP16-activator, GCN5, and Brd2 are still recruited to the transcription site but the chromatin does not decondense. Conclusions/Significance This study demonstrates that a strong activator can rapidly overcome the condensed chromatin structure of an inactive transcription site and supercede the expected requirement for regulatory events to proceed in a temporally defined order. Additionally, activator strength determines the number of cells in which transcription is induced as well as the extent of chromatin decondensation. As chromatin decondensation is significantly reduced after α-amanitin pre-treatment, despite the recruitment of transcriptional activation factors, this provides further evidence that transcription drives large-scale chromatin decondensation. PMID:20422051

  19. [Effect of retroauricular galvanic stimulation on the central vestibular system--immunohistochemical evaluation of GABA].

    PubMed

    Okami, K; Sekitani, T

    1990-03-01

    The changes of the neurotransmitter (GABA) distribution in the brain stem of the rats by retroauricular galvanic stimulation were investigated using immunohistochemical method. Twenty-one rats were divided into two groups: the control group which received no galvanic stimulation, and the galvanically stimulated group which received anodal galvanic stimulation (unipolar monoauricular, 5 mA in intensity, 500 msec of duration, 1 Hz in frequency) for 30 minutes. The specimens obtained as usual strict procedure for histological investigation were stained immunohistochemically using antisera against GABA. The results were as follows: 1. In the control group, GABA-like immunoreactivity was observed in all four main vestibular nuclei. In the superior, medial, and descending vestibular nuclei GABA-like immunoreactivity was found in the small cells and the terminals. Giant cells in the lateral vestibular nucleus were surrounded by GABA immunoreactive terminals. 2. In the galvanically stimulated group GABA-like immunoreactivity showed recognizable laterality in the lateral vestibular nucleus where GABA-like immunoreactivity surrounding giant cells showed more intensive on the side ipsilateral to the stimulation compared with the opposite side. On the other hand GABA-like immunoreactivity showed no laterality in the superior, medial, and descending vestibular nuclei. 3. It can be concluded that the retroauricular galvanic stimulation cause some changes in the inhibitory activity of the lateral vestibulo-spinal tract and of the spinal motor neuron.

  20. Immunohistochemical localization of caffeine in young Camellia sinensis (L.) O. Kuntze (tea) leaves.

    PubMed

    van Breda, Shane V; van der Merwe, Chris F; Robbertse, Hannes; Apostolides, Zeno

    2013-03-01

    The anatomical localization of caffeine within young Camellia sinensis leaves was investigated using immunohistochemical methods and confocal scanning laser microscopy. Preliminary fixation experiments were conducted with young C. sinensis leaves to determine which fixation procedure retained caffeine the best as determined by high-performance liquid chromatography analysis. High pressure freezing, freeze substitution, and embedding in resin was deemed the best protocol as it retained most of the caffeine and allowed for the samples to be sectioned with ease. Immunohistochemical localization with primary anti-caffeine antibodies and conjugated secondary antibodies on leaf sections proved at the tissue level that caffeine was localized and accumulated within vascular bundles, mainly the precursor phloem. With the use of a pressure bomb, xylem sap was collected using a micro syringe. The xylem sap was analyzed by thin-layer chromatography and the presence of caffeine was determined. We hypothesize that caffeine is synthesized in the chloroplasts of photosynthetic cells and transported to vascular bundles where it acts as a chemical defense against various pathogens and predators. Complex formation of caffeine with chlorogenic acid is also discussed as this may also help explain caffeine's localization.

  1. Immunohistochemical localization of caffeine in young Camellia sinensis (L.) O. Kuntze (tea) leaves.

    PubMed

    van Breda, Shane V; van der Merwe, Chris F; Robbertse, Hannes; Apostolides, Zeno

    2013-03-01

    The anatomical localization of caffeine within young Camellia sinensis leaves was investigated using immunohistochemical methods and confocal scanning laser microscopy. Preliminary fixation experiments were conducted with young C. sinensis leaves to determine which fixation procedure retained caffeine the best as determined by high-performance liquid chromatography analysis. High pressure freezing, freeze substitution, and embedding in resin was deemed the best protocol as it retained most of the caffeine and allowed for the samples to be sectioned with ease. Immunohistochemical localization with primary anti-caffeine antibodies and conjugated secondary antibodies on leaf sections proved at the tissue level that caffeine was localized and accumulated within vascular bundles, mainly the precursor phloem. With the use of a pressure bomb, xylem sap was collected using a micro syringe. The xylem sap was analyzed by thin-layer chromatography and the presence of caffeine was determined. We hypothesize that caffeine is synthesized in the chloroplasts of photosynthetic cells and transported to vascular bundles where it acts as a chemical defense against various pathogens and predators. Complex formation of caffeine with chlorogenic acid is also discussed as this may also help explain caffeine's localization. PMID:23143222

  2. Plasmacytoid myoepithelioma of minor salivary glands: report of case with emphasis in the immunohistochemical findings

    PubMed Central

    2011-01-01

    Myoepithelioma is a rare benign tumor of the salivary glands and is usually seen in the parotid gland and the minor salivary glands. It was once considered to be a type of pleomorphic adenoma (PA), but myoepitheliomas are today believed to be relatively aggressive tumors. Myoepitheliomas are most common in young adults between the ages of 30 and 50 and there are very few cases reported in individuals less than 18 years of age. We report a case of myoepithelioma located in the hard palate in a 15-year-old Brazilian male. The tumor was composed of plasmacytoid myoepithelial cells. An analysis of the immunohistochemical profile of the tumor cells showed positivity for vimentin, S-100 protein, and glial fibrillary acidic protein (GFAP), but not for smooth muscle actin (α-SMA) and cytokeratin 14 (CK14). We report this case because of the rarity of this tumor, especially in adolescents. We also discuss the histological parameters of the differential diagnosis of this tumor as well as its immunohistochemical profile. PMID:22152025

  3. The distinction of bronchogenic cyst from metastatic testicular teratoma: a light microscopic and immunohistochemical study.

    PubMed

    Roma, Andres; Varsegi, Mary; Magi-Galluzzi, Cristina; Ulbright, Thomas; Zhou, Ming

    2008-08-01

    Bronchogenic cyst (BC), a rare developmental abnormality of the primitive foregut and usually found above the diaphragm, can occur in the retroperitoneum and other unusual locations where distinction from metastatic teratoma is essential. We studied 22 BCs and 34 teratomas metastatic to the retroperitoneum in men. Of 22 BCs, 17 (77%) showed well or moderate architectural differentiation toward normal tracheobronchial structures, whereas none of the teratomas did. In addition, 20 (91%) of the BCs had respiratory-type epithelium and none had enteric-type epithelium. Immature elements, atypia, and tumor necrosis were absent in BCs. All BCs expressed cytokeratin (CK) 7, and none expressed CDX-2 in immunohistochemical analysis. Teratomas had mixed enteric and respiratory-type epithelium. The majority of the glands were positive for CK7 (33 [97%]), CK20 (34 [100%]), CDX-2 (34 [100%]), and thyroid transcription factor (TTF)-1 (17 [50%]). Coexpression of CDX-2 and TTF-1 was found in 11 teratomas (32%) but none of the BCs. BCs can be reliably distinguished from teratomas based on the gross, histologic, and immunohistochemical features.

  4. Non-sebaceous lymphadenoma of the salivary gland: case report with immunohistochemical investigation.

    PubMed

    Yang, Shaodong; Chen, Xinming; Wang, Li; Zhang, Jiali

    2007-05-01

    Non-sebaceous lymphadenoma (NSL) is a rare, recently described, benign salivary gland tumor characterized by a dense lymphoid infiltrate and absence of sebaceous differentiation. To our knowledge, only seven previous cases have been reported. In this paper, we describe an additional example of NSL along with an extensive analysis of its keratin (CK) profile. The patient was a 50-year-old woman presenting with a slowly growing painless mass in the right parotid gland. The tumor was encapsulated and measured 3 x 2 x 2 cm. Microscopically, the tumor comprised islands of epithelial cells with centrally located duct-like structures within a dense lymphoid stroma. Immunohistochemically, the tumor regularly expressed CKs 7, 8/18, and 19, which are typical for columnar differentiation and CKs 17 and 5/6, which are most typically expressed in basal cells of complex epithelia. CK14 was only expressed in rare scattered cells and eventually in groups of cells. The expression of CK10/13, which correlates with squamous differentiation, was negative. Additionally, immunostaining for smooth muscle actin, vimentin, and S-100 was also performed. The immunohistochemical findings in the neoplastic epithelial component of our case suggest a differentiation of "intercalated duct phenotype" without myoepithelial cell participation.

  5. Intruder Activity Analysis under Unreliable Sensor Networks

    SciTech Connect

    Tae-Sic Yoo; Humberto E. Garcia

    2007-09-01

    This paper addresses the problem of counting intruder activities within a monitored domain by a sensor network. The deployed sensors are unreliable. We characterize imperfect sensors with misdetection and false-alarm probabilities. We model intruder activities with Markov Chains. A set of Hidden Markov Models (HMM) models the imperfect sensors and intruder activities to be monitored. A novel sequential change detection/isolation algorithm is developed to detect and isolate a change from an HMM representing no intruder activity to another HMM representing some intruder activities. Procedures for estimating the entry time and the trace of intruder activities are developed. A domain monitoring example is given to illustrate the presented concepts and computational procedures.

  6. Metaplastic carcinoma of the breast: an immunohistochemical study

    PubMed Central

    2014-01-01

    Background Metaplastic breast carcinoma is a rare entity of breast cancer expressing epithelial and/or mesenchymal tissue within the same tumor. The aim of this study is to evaluate the clinicopathological features of metaplastic breast carcinoma and to confirm the triple negative, basal-like and/or luminal phenotype of this type of tumor by using immunohistochemical staining. Methods Seven cases of MBC were evaluated for clinico-pathological features including follow up data. Cases were studied immunohistochemically by CK-Pan, Vimentin, ER, PR, HER2, basal markers (CK5/6, p63, EGFR, SMA and S-100), luminal cytokeratins (CK8, CK18 and CK19), markers for syncytial cells (β-HCG and PLAP), as well as prognostic markers (p53, ki-67 and calretinin). Results The mean age of the patients was 36 years. Three cases showed choriocarcinomatous features. All of our cases were negative for ER, PR and HER2. Six out of the 7 cases showed basal-like differentiation by demonstrating positivity with at least one of the basal/myoepithelial markers. Also 6 out of the 7 cases expressed luminal type cytokeratins (CK8, CK18 and/or CK19). P53 was positive in 3 cases, ki-67 was strongly expressed in only one case, while calretinin was expressed in 6 cases. Conclusion Metaplastic breast carcinoma presents in our population at a younger age group than other international studies. All cases are categorized immunohistochemically under the triple negative group of breast cancer and 86% of them exhibited basal-like and luminal phenotype. Majority of cases developed local recurrence and distant metastasis in a relatively short period of time. Virtual Slides The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1101289295115804 PMID:25030022

  7. C4d staining as immunohistochemical marker in inflammatory myopathies.

    PubMed

    Pytel, Peter

    2014-10-01

    The diagnosis of an inflammatory myopathy is often established based on basic histologic studies. Additional immunohistochemical studies are sometimes required to support the diagnosis and the classification of inflammatory myopathies. Staining for major histocompatibility complex 1 (MHC1) often shows increased sarcolemmal labeling in inflammatory myopathies. Endomysial capillary staining C5b-9 (membrane attack complex) is a feature that is reported as frequently associated with dermatomyositis. Immunohistochemical staining for C4d is widely used for various applications including the assessment of antibody-mediated rejection after solid organ transplantation. In the context of dermatomyositis, C4d staining has been described in skin biopsies but not in muscle biopsies. A total of 32 muscle biopsy specimens were examined. The hematoxylin and eosin-stained slides were reviewed, and immunohistochemical studies for MHC1, C5b-9, and C4d were conducted. The staining observed for C5b-9 and C4d was compared. Overall, the staining pattern for C4d mirrored the one observed for C5b-9 in the examined muscle biopsy specimens. There was high and statistically significant (P<0.0001) correlation between the staining seen with these 2 antibodies. Both antibodies labeled the cytoplasm of degenerating necrotic myofibers. In addition, both antibodies showed distinct endomysial capillary labeling in a subset of dermatomyositis. Areas with perifascicular atrophy often exhibited the most prominent vascular labeling for C4d and C5b-9. In conclusion, C4d and C5b-9 show similar expression patterns in muscle biopsies of patients with inflammatory myopathies and both highlight the presence of vascular labeling associated with dermatomyositis. C4d antibodies are widely used and may offer an alternative for C5b-9 staining.

  8. High-Throughput Analysis of Enzyme Activities

    SciTech Connect

    Lu, Guoxin

    2007-01-01

    High-throughput screening (HTS) techniques have been applied to many research fields nowadays. Robot microarray printing technique and automation microtiter handling technique allows HTS performing in both heterogeneous and homogeneous formats, with minimal sample required for each assay element. In this dissertation, new HTS techniques for enzyme activity analysis were developed. First, patterns of immobilized enzyme on nylon screen were detected by multiplexed capillary system. The imaging resolution is limited by the outer diameter of the capillaries. In order to get finer images, capillaries with smaller outer diameters can be used to form the imaging probe. Application of capillary electrophoresis allows separation of the product from the substrate in the reaction mixture, so that the product doesn't have to have different optical properties with the substrate. UV absorption detection allows almost universal detection for organic molecules. Thus, no modifications of either the substrate or the product molecules are necessary. This technique has the potential to be used in screening of local distribution variations of specific bio-molecules in a tissue or in screening of multiple immobilized catalysts. Another high-throughput screening technique is developed by directly monitoring the light intensity of the immobilized-catalyst surface using a scientific charge-coupled device (CCD). Briefly, the surface of enzyme microarray is focused onto a scientific CCD using an objective lens. By carefully choosing the detection wavelength, generation of product on an enzyme spot can be seen by the CCD. Analyzing the light intensity change over time on an enzyme spot can give information of reaction rate. The same microarray can be used for many times. Thus, high-throughput kinetic studies of hundreds of catalytic reactions are made possible. At last, we studied the fluorescence emission spectra of ADP and obtained the detection limits for ADP under three different

  9. Cartilaginous choristoma of the tongue with an immunohistochemical study.

    PubMed

    Pereira, Gregório Wrublevski; Pereira, Volnei David; Pereira Junior, José Antonio de Castro; da Silva, Rosemeri Maurici

    2012-12-06

    By definition, choristomas are normal tissues found in anomalous topography. The cartilaginous features of these lesions are rare in the soft tissues of the oral cavity. The majority of cartilaginous choristomas of the tongue--the primary site of emergence of the oropharynx--are associated with adipose, fibrous or bone tissues--apart from that, only a few of these were confirmed by an immunohistochemical study. The neoplasm exclusively composed of chondromatous tissue is extremely rare in the tongue. This paper reports the clinical, surgical and pathological characteristics of a cartilaginous choristoma of the tongue diagnosed in a 64-year-old woman.

  10. Morphology and Immunohistochemical Phenotype of the Thymus in Secondary Immunodeficiency.

    PubMed

    Struchko, G Yu; Merkulova, L M; Moskvichev, E V; Kostrova, O Yu; Mikhailova, M N; Drandrova, E G

    2015-10-01

    The thymus of outbred male rats 5 months after splenectomy (experimental secondary immunodeficiency) was studied by common histological and immunohistochemical methods using monoclonal and polyclonal antibodies to CD3, CD30, CD68, synaptophysin, to S100, p53, bcl-2, and Ki-67 proteins. Removal of the spleen led to acute involution of the thymic parenchyma, which was replaced by the adipose tissue and was associated with restructuring of the thymopoietic and nonthymopoietic components of the gland, changes in cellular composition and antigenic phenotype of the lobular cortical and medullary matter, and by reduction of cell proliferation.

  11. Massive retinal gliosis: An unusual case with immunohistochemical study

    PubMed Central

    Deshmukh, Sanjay D; Ashturkar, Amrut V; Babanagare, Shridhar V; Gokhale, Suvarna K; Deshpande, Anand A

    2011-01-01

    Massive retinal gliosis (MRG) is a rare, benign intraocular condition that results from the proliferation of well-differentiated glial cells. Immunohistochemically, these cells show positivity for glial fibrillary acid protein (GFAP), neuron specific enolase (NSE), and S-100 protein. We encountered a case of a 45-year-old female with loss of vision in the left eye. She had a history of trauma to that eye two years ago. Enucleation was carried out, because malignancy was suspected due to retinal calcification. On the basis of light microscopy and immunohistochemistry (IHC) performed on the enucleated eye, it was diagnosed as massive retinal gliosis. PMID:21586853

  12. Karyotype Analysis Activity: A Constructivist Learning Design

    ERIC Educational Resources Information Center

    Ahmed, Noveera T.

    2015-01-01

    This classroom activity is based on a constructivist learning design and engages students in physically constructing a karyotype of three mock patients. Students then diagnose the chromosomal aneuploidy based on the karyotype, list the symptoms associated with the disorder, and discuss the implications of the diagnosis. This activity is targeted…

  13. [Immunohistochemical study of CART-peptide in striato-nigral projections at dopamine loss].

    PubMed

    Romanova, I V; Chesnokova, A Iu; Mikhrina, A L

    2012-08-01

    The increase of CART-peptide optical density was found immunohistochemically in nucleus accumbens neurons and in their terminals in substantia nigra in Wistar rats after 28% reduction of dopaminergic neurons in a substantia nigra (in the model of lactacystin induced proteo some disfunction). At the same time after in vitro incubation of nigro-accumbal brain slice with AMPT (alpha-methyl-paratirosine--dopamine inhibitor) for 4 h the reduction of tyrosine hydroxylase optical density (the enzyme limiting dopamine synthesis) in substantia nigr neurons was found and optical density of CART-peptide in nucleus accumbens and substantia nigra was also revealed. In both experiments data about activation of CARTergic neurons in stria to-nigral projections testifies on participation of CART-peptide in compensatory brain mechanisms at dopamine loss and its role as modulator of dopaminergic brain neurons functional activity.

  14. Comprehensive immunohistochemical analysis of Her-2/neu oncoprotein overexpression in breast cancer: HercepTest (Dako) for manual testing and Her-2/neuTest 4B5 (Ventana) for Ventana BenchMark automatic staining system with correlation to results of fluorescence in situ hybridization (FISH).

    PubMed

    Mayr, Doris; Heim, Sibylle; Werhan, Cedric; Zeindl-Eberhart, Evelyn; Kirchner, Thomas

    2009-03-01

    Overexpression of Her-2/neu-oncoprotein is used as marker for Herceptin therapy. To investigate the sensitivity and specificity of automatic immunohistochemistry (Benchmark, Ventana), we compared the results to the manual testing (Dako) in 130 breast carcinomas and validated the results by fluorescence in situ hybridization (FISH). Manual and automatic immunohistochemistry of Her-2/neu-oncoprotein using two different antibodies (HercepTest, Her-2/neuTest 4B5) was analyzed. FISH was performed in all cases with uncertain or strong overexpression in either immunohistochemical stainings or with different immunohistochemical results. Same immunohistochemical results were seen in 73.8%. Two cases with overexpression, detected with Her-2/neuTest 4B5 and confirmed by FISH, showed no overexpression using HercepTest. From 21 cases with 2+ by Her-2/neuTest 4B5, 15 cases had no gene amplification (two of them with 3+ HercepTest); three cases showed a gene amplification (one of them with failing overexpression by HercepTest); two other cases were polysomic; one could not be analyzed. Ventana immunohistochemistry seems to be of same reliability like Dako with a little better concordance to FISH in our study.

  15. Oral soft-tissue angiolipoma: report of two cases of rare oral lipomatous lesion with emphasis on morphological and immunohistochemical features.

    PubMed

    Silva-Junior, Geraldo O; Picciani, Bruna L; Costa, Raphael C; Barbosa, Saulo M; Silvares, Marcelo G; Souza, Ramiro B; Cantisano, Marília H; Pires, Fabio R

    2013-03-01

    Oral angiolipomas are exceedingly rare and little is known about their morphological and etiological features. Here, we report two cases of oral angiolipoma and discuss their clinicopathological and immunohistochemical features, focusing on endothelial markers. Both lesions presented mature adipocytes interspersed by small blood vessels containing fibrin thrombi. Immunohistochemical analysis showed numerous mast cells and expression of CD34, vascular endothelial growth factor, intercellular adhesion molecule-1, interferon-γ and interleukin 6 in most endothelial and stromal cells. Mast cell-endothelial cell interaction may be responsible for the reactive or neoplastic origin of the vascular proliferation of these entities.

  16. Histological and immunohistochemical study of Wischnewsky spots in fatal hypothermia.

    PubMed

    Tsokos, Michael; Rothschild, Markus A; Madea, Burkhard; Rie, Manfred; Sperhake, Jan P

    2006-03-01

    Wischnewsky spots in the gastric mucosa are considered an important finding for the diagnosis of hypothermia-related deaths. In the present prospective histological and immunohistochemical investigation, 14 cases of fatal hypothermia presenting Wischnewsky spots at autopsy were studied. Macromorphologically, the lesions, varying in diameter from 0.1 to 0.4 cm, had a blackish-brownish color and appeared partly lofty, especially on the apex of gastric folds. Histologically, no erosions or ulcers were observed in the gastric mucosa. In some cases, hemorrhages in conjunction with infarctions of the mucosa were observed in the mucosal glands. Those regions, however, did not represent the lesions visible as Wischnewsky spots at the macroscopical level. Immunohistochemical stains were done with a specific antibody against hemoglobin (Dako, Glostrup, Denmark). Wischnewsky spots expressed immunopositivity with antihemoglobin. Concerning the pathogenesis and underlying pathophysiologic mechanisms contributing to the development of Wischnewsky spots, we hypothesize that cooling of the body in the setting of cold ambient temperatures primarily leads to circumscribed hemorrhages of the gastric glands in vivo or in the agonal period, respectively. Subsequently, due to autolysis, erythrocytes are destroyed and hemoglobin is released. Following exposure to gastric acid, hemoglobin is hematinized, leading to the typical blackish-brownish appearance of Wischnewsky spots seen at gross examination. Wischnewsky spots are not equivalent to erosions in terms of histopathological diagnosis but rather represent epiphenomena generated in vivo or in the agonal period of fatal hypothermia. PMID:16501354

  17. Pleomorphic Carcinoma of the Colon: Morphological and Immunohistochemical Findings

    PubMed Central

    Branca, Giovanni; Barresi, Valeria; Ieni, Antonio; Irato, Eleonora; Caruso, Rosario Alberto

    2016-01-01

    Pleomorphic carcinoma is an aggressive neoplasm defined by the World Health Organization (WHO) as a poorly differentiated (squamous cell carcinoma or adenocarcinoma) or undifferentiated carcinoma in which at least 10% spindle and/or giant cells are identified, or as a carcinoma constituted purely of spindle and giant cells. Although this entity has initially been shown in the lung, it has been described also in extrapulmonary locations, with only one report for a colonic site. A 65-year-old woman developed a caecal tumour. Gross examination revealed an endophytic/ulcerative mass 7 cm in length. Microscopically, the tumour was a poorly differentiated adenocarcinoma with a pleomorphic component that occupied more than 10% of the specimen. The tumour shared these histopathological findings with pulmonary giant cell carcinoma but differed in other clinicopathological features such as a pushing growth pattern, stage pT3N1, and an uneventful outcome 24 months after operation. The pleomorphic component showed morphological and immunohistochemical features compatible with mitotic catastrophe, a non-apoptotic cell death occurring in cycling cells after aberrant mitosis. These features included multinucleation, micronucleation, atypical mitoses, foci of geographic necrosis, as well as immunohistochemical overexpression of p53 and Ki-67. The interpretation of the pleomorphic component as morphological expression of mitotic catastrophe may be useful in comprehending the pathogenesis of this rare neoplasm, and it may have practical implications as a potential cancer therapeutic target. PMID:27462191

  18. Radiological, histological and immunohistochemical evaluation of periapical inflammatory lesions.

    PubMed

    Berar, Antonela Marcela; Bondor, Cosmina Ioana; Matroş, LuminiŢa; Câmpian, Radu Septimiu

    2016-01-01

    The loss of teeth is largely caused by supporting tissue damage, because of bacterial invasion from the infected root canals. Sixty patients with periapical lesions (PLs) of endodontic origin were included in the study. Clinical and radiological examination was performed. Periapical radiographs were analyzed by two independent observers to determine the size and severity of PLs, using Periapical Index (PAI) scores. The tissue samples collected by periapical curettage during apicoectomy or after dental extractions by alveolar curettage were histologically and immunohistochemically analyzed. The PLs were histologically diagnosed as: periapical granulomas (PGs), granulomas with cystic potential and radicular cysts (RCs) with various degrees of inflammation. Capillary density was evaluated using the angiogenic index after immunohistochemical staining with CD34 monoclonal antibody. A statistically significant correlation was observed between PAI scores and the size of the lesions. 68.33% of cases were PGs, 18.33% PGs with cystic potential and 18.33% RCs with different degrees of inflammation. Seventy-five percent PLs had an angiogenic index 1 and 25% had an angiogenic index 2. Statistically significant differences were obtained between the angiogenic index and lesion size (p<0.05). Capillary density within PLs did not influence the severity scores of lesions detected on radiographs. The angiogenic index appeared not to be associated with the histological lesion type and the intensity of inflammation, but was more likely correlated with the degree of granulation tissue maturation and the size of PLs. PMID:27516014

  19. Immunohistochemical patterns in the differential diagnosis of rhinopharyngeal granulocytic sarcoma

    PubMed Central

    Cantone, Elena; Cavaliere, Michele; Di Lullo, Antonella Miriam; Guadagno, Elia; Iengo, Maurizio

    2016-01-01

    Granulocytic sarcoma (GS) is a rare extramedullary manifestation of acute myeloid leukemia (AML). GS may develop simultaneously to AML or as a relapse of leukemia, particularly following allogeneic hematopoietic stem cell transplant. Subperiosteal bone, lymph nodes and skin are commonly involved, whereas rhinopharyngeal involvement is less common, with only 14 cases reported in the literature. Due to its rarity, rhinopharyngeal GS may lead to diagnostic pitfalls, particularly when it is poorly differentiated or is without concomitant marrow involvement. Thus, immunohistochemical findings play a key role in diagnosis. The current report describes a case of a 53-year-old female suffering from rhinopharyngeal GS and with a history of AML treated with chemotherapy and radiotherapy, focusing on the importance of the immunohistochemical pattern to assess the right diagnosis. Recent studies have demonstrated that the immunophenotype is of utmost importance for the diagnosis of GS. The high expression of myeloperoxidase (MPO) is common in GS; however, ~30% of GSs do not contain MPO. Therefore, the presence of other markers is required to confirm the diagnosis of GS.

  20. Immunohistochemical expression of type VI collagen in superficial fibromatoses.

    PubMed

    Magro, G; Colombatti, A; Lanzafame, S

    1995-10-01

    The expression of type VI collagen was studied immunohistochemically in 26 cases of superficial fibromatoses (palmar, plantar and penile) using an immunoperoxidase method for light microscopic visualization. The polyclonal antibody against type VI collagen used in this study was isolated from human placenta and its specifity was tested by immunoblotting assay. All cases consisted of multiple nodules showing a variable degree of cellularity and fibrosis. Depending on the predominant histological appearance of these nodules, each case was assigned to the three following phases: proliferative, involutional and residual. Morphologically normal palmar and plantar aponeuroses were included as controls. Immunohistochemical findings showed that type VI collagen was present as longitudinal thin fibers in normal palmar and plantar aponeuroses. A differential expression of this collagen was found in the different stages of superficial fibromatoses. Type VI collagen was markedly expressed as a distinct fibrillar network in the extracellular matrix (ECM) surrounding proliferating stromal cells in proliferative and involutional phases. Its expression completely disappeared from the connective tissue undergoing fibrotic transformation during involutional and residual phases. The results of the present study suggest that type VI collagen is an extracellular marker of stromal tissue proliferation and is involved in the early phases of tissue remodelling occurring in the superficial fibromatoses.

  1. Collagenase-3 expression in periapical lesions: an immunohistochemical study.

    PubMed

    Bhalla, G; Astekar, M S; Ramesh, G; Kaur, P; Sowmya, G V

    2014-08-01

    Collagenase-3 (matrix metalloproteinase-13) is a metalloproteinase (MMP) that is associated with bone lesions and exhibits variable expression patterns in odontogenic cysts; it may play a role in regulating focal proliferation and maturation of jaw cyst epithelium. We studied the localization, staining intensity and distribution of collagenase-3 in 13 periapical granulomas with epithelium, 16 periapical granulomas without epithelium and 10 radicular cysts using archived formalin fixed, paraffin embedded tissues. A monoclonal antibody against human collagenase-3 was used to evaluate its expression. Immunohistochemical staining intensities of collagenase-3 in all periapical lesions were (-), 4 (10%); (+), 1 (3%); (++), 22 (56%) and (+++), 12 (31%); differences were not statistically significant. Immunohistochemical distribution of collagenase-3 in epithelial cells was (-), 17 (44%); (+), 17 (44%); (++), 5 (13%); in fibroblasts it was (-), 8 (20%); (+), 23 (59%); (++), 8 (21%); in plasma cells it was (-), 7 (18%); (+), 22 (56%); (++), 10 (26%); in macrophages it was (-), 7 (18%); (+), and 15 (38%); and (++), 17 (44%). Statistically significant differences were found in epithelial cells (p = 0.00) and fibroblasts (p = 0.02), whereas differences were not statistically significant for plasma cells and macrophages. Collagenase-3 may play a role in the conversion of a periapical granuloma with epithelium to radicular cyst. MMP's influence not only epithelial rest cell migration, but also invasion of various stromal cells into granulomatous tissue.

  2. Oral mucoceles: a clinical, histopathological and immunohistochemical study.

    PubMed

    Conceição, Jamile Gomes; Gurgel, Clarissa Araújo; Ramos, Eduardo Antônio Gonçalves; De Aquino Xavier, Flávia Caló; Schlaepfer-Sales, Caroline Brandi; Cangussu, Maria Cristina Teixeira; Cury, Patrícia Ramos; Ramalho, Luciana Maria Pedreira; Dos Santos, Jean Nunes

    2014-01-01

    The aim of study was to evaluate the clinicopathological features of oral mucoceles and the immunohistochemical expression of cellular and extracellular matrix components in these lesions. One hundred cases of oral mucoceles were examined for clinicopathological features. The expression of mast cell tryptase, CD68, MMP-1 (matrix metalloproteinase-1), MMP-9 (matrix metalloproteinase-9) and CD34 was investigated immunohistochemically in 32 cases. The lesions arose as nodules or blisters of variable color. The mean age was 23.2 years and a higher male frequency was observed. The most common locations were the lower lip (92%), followed by the floor of the mouth (7%), and palate (1%). The lesion size ranged from 0.4 to 3.0cm. Unusual histopathological findings as superficial mucoceles (n=16, 16%), pseudopapillary projections (n=3, 3%), epithelioid histiocytes (n=4, 4%), multinucleated giant cells (n=1, 1%) and myxoglobulosis (n=9, 9%) were also seen. Mast cells and CD68-positive macrophages, MMP-1, MMP-9 and CD34-positive blood vessels were seen in all cases. A significant association was seen between mast cells and MMP-1 (p=0.03) and between macrophages and MMP-1 (p=0.01). This study provided important insight into the demographic and histopathological occurrence of oral mucoceles. The tissue remodeling seen in these lesions mainly involved the migration and interaction of mast cells, macrophages and MMP-1. PMID:23726142

  3. Immunohistochemical and Molecular Characterization of the Human Periosteum

    PubMed Central

    Frey, Sönke Percy; Jansen, Hendrik; Doht, Stefanie; Filgueira, Luis; Zellweger, Rene

    2013-01-01

    Purpose. The aim of the present study was to characterize the cell of the human periosteum using immunohistological and molecular methods. Methods. Phenotypic properties and the distribution of the cells within the different layers were investigated with immunohistochemical staining techniques and RT-PCR, focussing on markers for stromal stem cells, osteoblasts, osteoclasts and immune cells. Results. Immunohistochemical results revealed that all stained cells were located in the cambium layer and that most cells were positive for vimentin. The majority of cells consisted of stromal stem cells and osteoblastic precursor cells. The density increased towards the deeper layers of the cambium. In addition, cells positive for markers of the osteoblast, chondrocyte, and osteoclast lineages were found. Interestingly, there were MHC class II-expressing immune cells suggesting the presence of dendritic cells. Using lineage-specific primer pairs RT-PCR confirmed the immunofluorescence microscopy results, supporting that human periosteum serves as a reservoir of stromal stem cells, as well as cells of the osteoblastic, and the chondroblastic lineage, osteoclasts, and dendritic cells. Conclusion. Our work elucidates the role of periosteum as a source of cells with a high regenerative capacity. Undifferentiated stromal stem cells as well as osteoblastic precursor cells are dominating in the cambium layer. A new outlook is given towards an immune response coming from the periosteum as MHC II positive immune cells were detected. PMID:23737713

  4. Characterization of equine hyalocytes: their immunohistochemical properties, morphologies and distribution.

    PubMed

    Sano, Yuto; Matsuda, Kazuya; Okamoto, Minoru; Takehana, Kazushige; Hirayama, Kazuko; Taniyama, Hiroyuki

    2016-07-01

    In horse, the characterizations of hyalocytes under the steady state are still unclear. Therefore, we investigated characterizations of hyalocytes in normal equine eyes by their immunohistochemical phenotype, histomorphology and distribution. Thirty-one eyes from 18 horses, divided into 4 groups (G) by age, were used: early (G1) and late gestation (G2) fetuses, 1- to 3-year-old (G3) and 8- to 24-year-old (G4) horses. Equine hyalocytes were histologically classified into 4 types, and they immunohistochemically expressed MHC II and CD163. Hyalocytes were detected on and/or around ciliary processes and pars plana in G2, G3 and G4, but were not located on retina and optic papilla. A significant increase in distribution was found between G2 and both G3 and G4, and the largest distribution was found at ciliary processes in these groups. Equine hyalocytes were characterized as residential ocular macrophage and MHC II antigen-bearing cell, accompanied by a pleomorphic appearance and located in the contiguous ciliary body. Our data provided characterizations of hyalocytes in normal equine eyes and may well contribute to improving the understanding of pathogenesis of equine ocular disease.

  5. Immunohistochemical patterns in the differential diagnosis of rhinopharyngeal granulocytic sarcoma

    PubMed Central

    Cantone, Elena; Cavaliere, Michele; Di Lullo, Antonella Miriam; Guadagno, Elia; Iengo, Maurizio

    2016-01-01

    Granulocytic sarcoma (GS) is a rare extramedullary manifestation of acute myeloid leukemia (AML). GS may develop simultaneously to AML or as a relapse of leukemia, particularly following allogeneic hematopoietic stem cell transplant. Subperiosteal bone, lymph nodes and skin are commonly involved, whereas rhinopharyngeal involvement is less common, with only 14 cases reported in the literature. Due to its rarity, rhinopharyngeal GS may lead to diagnostic pitfalls, particularly when it is poorly differentiated or is without concomitant marrow involvement. Thus, immunohistochemical findings play a key role in diagnosis. The current report describes a case of a 53-year-old female suffering from rhinopharyngeal GS and with a history of AML treated with chemotherapy and radiotherapy, focusing on the importance of the immunohistochemical pattern to assess the right diagnosis. Recent studies have demonstrated that the immunophenotype is of utmost importance for the diagnosis of GS. The high expression of myeloperoxidase (MPO) is common in GS; however, ~30% of GSs do not contain MPO. Therefore, the presence of other markers is required to confirm the diagnosis of GS. PMID:27698857

  6. Characterization of equine hyalocytes: their immunohistochemical properties, morphologies and distribution

    PubMed Central

    SANO, Yuto; MATSUDA, Kazuya; OKAMOTO, Minoru; TAKEHANA, Kazushige; HIRAYAMA, Kazuko; TANIYAMA, Hiroyuki

    2016-01-01

    In horse, the characterizations of hyalocytes under the steady state are still unclear. Therefore, we investigated characterizations of hyalocytes in normal equine eyes by their immunohistochemical phenotype, histomorphology and distribution. Thirty-one eyes from 18 horses, divided into 4 groups (G) by age, were used: early (G1) and late gestation (G2) fetuses, 1- to 3-year-old (G3) and 8- to 24-year-old (G4) horses. Equine hyalocytes were histologically classified into 4 types, and they immunohistochemically expressed MHC II and CD163. Hyalocytes were detected on and/or around ciliary processes and pars plana in G2, G3 and G4, but were not located on retina and optic papilla. A significant increase in distribution was found between G2 and both G3 and G4, and the largest distribution was found at ciliary processes in these groups. Equine hyalocytes were characterized as residential ocular macrophage and MHC II antigen-bearing cell, accompanied by a pleomorphic appearance and located in the contiguous ciliary body. Our data provided characterizations of hyalocytes in normal equine eyes and may well contribute to improving the understanding of pathogenesis of equine ocular disease. PMID:26888584

  7. Occurrence of trichoepithelioma in a cat: Histopathologic and immunohistochemical study

    PubMed Central

    Tavasoli, Abbas; Golshahi, Hannaneh; Rad, Maryam Nobakht; Taymouri, Afra

    2013-01-01

    Trichoepitheliomas are benign follicular appendage tumors with differentiation to all three segments of the hair follicle. A 2 years old female domestic short hair cat presented with a mass on the tail. The mass was surgically excised and for histopathologic and immunohistochemical studies, was sent to Department of Pathology. Histologically, the tumor was encapsulated and consisted of many islands of follicular epithelium and also cysts structures which varied in size and shape. The cells of epithelium islands were round to oval and had variable amounts of slightly, eosinophilic cytoplasm and euchromatic nuclei. The cystic structures were lined by a complex layer of squamous epithelium. Often, cells under went an abrupt transition between basal layers and keratinization without the development of a granular cell layer. No tendency of malignancy was seen in this case. According to mentioned characteristics, trichoepithelioma was diagnosed. By immunohistochemical study it was confirmed that this tumor had epithelial origin because squamous tumor cells reacted with the pan-cytokeratin antibody. The expression of β-catenin was predominately cytoplasmic and also together with numerous positive nuclei but membranous expression was inconsistenet. Distribution of neoplastic cells with β-catenin expression was more than 75% and labeling intensity was strong in both cytoplasm and nuclei. According to author's knowledge, this is the first report of trichoepithelioma in cat in Iran and also investigation of β-catenin expression in feline trichoepithelioma in veterinary literature. PMID:23646307

  8. Histiocytoid breast carcinoma: a case report showing immunohistochemical profiles.

    PubMed

    Li, Peifeng; Zheng, Jinfeng; Zhang, Tingguo; Cao, Ruixue; Chen, Xin; Geng, Ming

    2013-01-01

    Histiocytoid breast carcinoma (HBC) is a rare type of breast cancer with a controversial histogenesis. Here we describe a case report of a 65-year old woman with HBC. The patient presented with two masses in the right breast. Histopathologically, the tumors consisted of a diffuse infiltration of large tumor cells and histological components of carcinoma in situ and atypical lobular hyperplasia were also observed. The infiltration pattern was similar to that of invasive lobular carcinoma with targetoid and Indian file arrangements. The invasive histiocytoid cells had finely granular, eosinophilic to vesicular cytoplasm and nuclei with a bland uniform appearance, a single small eosinophilic nucleolus and finely granular chromatin. We compared the immunohistochemical profiles of 17 breast cancer markers between invasive carcinoma, carcinoma in situ, atypical lobular hyperplasia and normal breast epithelium. Although they all shared the same reactivity for many of the proteins, they exhibited differences in GCDFP-15, E-cadherin, P120, CEA, HER-2, ER and PR expression, and these are discussed. This is the first case study of two HBC masses occurring in one breast simultaneously. By analyzing and comparing their morphologic characteristics and spectrum of immunohistochemical expression, our study supports the view that HBC is a variant of lobular carcinoma and our findings may assist in future diagnoses of HBC. PMID:24228128

  9. Molecular and Immunohistochemical Study of Platelet Derived Growth Factor Receptor Alpha in KIT Negative Gastrointestinal Stromal Tumors; the First Report from Iran

    PubMed Central

    Geramizadeh, Bita; Jowkar, Zahra; Mousavi, Seyed-Javad

    2016-01-01

    BACKGROUND Gastrointestinal stromal tumors (GISTs) are potentially malignant tumors; however their behavior and response to treatment is dependent on the type of mutation and immunohistochemical expression of antigens. It is recommended to perform routine molecular and immunohistochemical tests in KIT and platelet derived growth factor receptor alpha (PDGFRA) molecules for making decision regarding targeted therapy and prediction of the behavior of the tumor. Objectives: There has been no study from Iran regarding the PDGFRA mutational analysis in GISTs. In this study, for the first time from Iran, we performed immunohistochemical and molecular analysis of PDGFRA molecule on GISTs. METHODS In a cross-sectional study during 7 years (2008-2014) on 50 untreated non-recurrent non-metastatic newly diagnosed GISTs, molecular analysis and immunohistochemical staining for PDGFRA were performed and findings were compared with different clinicopathological characteristics.. RESULTS During the 7 years, 50 cases of GISTs according to the above mentioned criteria were found. 17 cases were negative for KIT mutation. Of them, 15 (30%) were positive for either exon 12 or 18 mutation of PDGFRA. These cases showed more epithelioid morphology and the number of mitotic figures were lower than PDFRA negative GISTs. Also according to the criteria for risk assessment, it seems that PDGFRA mutant GISTs are rarely in the high risk category. CONCLUSION PDGFRA mutant GISTs are common in Iranian population and it is recommended to perform mutation analysis for PDGFRA in every GIST with wild type KIT and epithelioid morphology. PMID:27698973

  10. Molecular and Immunohistochemical Study of Platelet Derived Growth Factor Receptor Alpha in KIT Negative Gastrointestinal Stromal Tumors; the First Report from Iran

    PubMed Central

    Geramizadeh, Bita; Jowkar, Zahra; Mousavi, Seyed-Javad

    2016-01-01

    BACKGROUND Gastrointestinal stromal tumors (GISTs) are potentially malignant tumors; however their behavior and response to treatment is dependent on the type of mutation and immunohistochemical expression of antigens. It is recommended to perform routine molecular and immunohistochemical tests in KIT and platelet derived growth factor receptor alpha (PDGFRA) molecules for making decision regarding targeted therapy and prediction of the behavior of the tumor. Objectives: There has been no study from Iran regarding the PDGFRA mutational analysis in GISTs. In this study, for the first time from Iran, we performed immunohistochemical and molecular analysis of PDGFRA molecule on GISTs. METHODS In a cross-sectional study during 7 years (2008-2014) on 50 untreated non-recurrent non-metastatic newly diagnosed GISTs, molecular analysis and immunohistochemical staining for PDGFRA were performed and findings were compared with different clinicopathological characteristics.. RESULTS During the 7 years, 50 cases of GISTs according to the above mentioned criteria were found. 17 cases were negative for KIT mutation. Of them, 15 (30%) were positive for either exon 12 or 18 mutation of PDGFRA. These cases showed more epithelioid morphology and the number of mitotic figures were lower than PDFRA negative GISTs. Also according to the criteria for risk assessment, it seems that PDGFRA mutant GISTs are rarely in the high risk category. CONCLUSION PDGFRA mutant GISTs are common in Iranian population and it is recommended to perform mutation analysis for PDGFRA in every GIST with wild type KIT and epithelioid morphology.

  11. Overview of MSFC's Applied Fluid Dynamics Analysis Group Activities

    NASA Technical Reports Server (NTRS)

    Garcia, Roberto; Griffin, Lisa; Williams, Robert

    2002-01-01

    This viewgraph report presents an overview of activities and accomplishments of NASA's Marshall Space Flight Center's Applied Fluid Dynamics Analysis Group. Expertise in this group focuses on high-fidelity fluids design and analysis with application to space shuttle propulsion and next generation launch technologies. Topics covered include: computational fluid dynamics research and goals, turbomachinery research and activities, nozzle research and activities, combustion devices, engine systems, MDA development and CFD process improvements.

  12. Acceleration of reverse analysis method using hyperbolic activation function

    NASA Astrophysics Data System (ADS)

    Pwasong, Augustine; Sathasivam, Saratha

    2015-10-01

    Hyperbolic activation function is examined for its ability to accelerate the performance of doing data mining by using a technique named as Reverse Analysis method. In this paper, we describe how Hopfield network perform better with hyperbolic activation function and able to induce logical rules from large database by using reverse analysis method: given the values of the connections of a network, we can hope to know what logical rules are entrenched in the database. We limit our analysis to Horn clauses.

  13. An Analysis of Solar Global Activity

    NASA Astrophysics Data System (ADS)

    Mouradian, Zadig

    2013-02-01

    This article proposes a unified observational model of solar activity based on sunspot number and the solar global activity in the rotation of the structures, both per 11-year cycle. The rotation rates show a variation of a half-century period and the same period is also associated to the sunspot amplitude variation. The global solar rotation interweaves with the observed global organisation of solar activity. An important role for this assembly is played by the Grand Cycle formed by the merging of five sunspot cycles: a forgotten discovery by R. Wolf. On the basis of these elements, the nature of the Dalton Minimum, the Maunder Minimum, the Gleissberg Cycle, and the Grand Minima are presented.

  14. Method for photon activation positron annihilation analysis

    DOEpatents

    Akers, Douglas W.

    2006-06-06

    A non-destructive testing method comprises providing a specimen having at least one positron emitter therein; determining a threshold energy for activating the positron emitter; and determining whether a half-life of the positron emitter is less than a selected half-life. If the half-life of the positron emitter is greater than or equal to the selected half-life, then activating the positron emitter by bombarding the specimen with photons having energies greater than the threshold energy and detecting gamma rays produced by annihilation of positrons in the specimen. If the half-life of the positron emitter is less then the selected half-life, then alternately activating the positron emitter by bombarding the specimen with photons having energies greater then the threshold energy and detecting gamma rays produced by positron annihilation within the specimen.

  15. Faculty Activity Analysis in the Universidad Tecnica Del Estado Campuses.

    ERIC Educational Resources Information Center

    Karadima, Oscar

    An analysis of academic activities of college faculty at the eight campuses of Chile's Universidad Tecnica del Estado was conducted. Activities were grouped into seven categories: direct teaching, indirect teaching, research, community services, faculty development, academic administration, and other activities. Following the narrative…

  16. Clinicopathological and immunohistochemical features of pulmonary artery sarcoma: A report of three cases and review of the literature

    PubMed Central

    CHEN, DONG; ZHU, GUANGFA; WANG, DIJIA; ZHANG, ZHE; FANG, WEI; QU, ZHENG

    2016-01-01

    Pulmonary artery sarcoma (PAS) is an extremely rare and highly malignant tumor that originates in the pulmonary artery. The majority of reported cases of PAS are confirmed by pathological examination subsequent to surgery or by autopsy. The present study reports the clinicopathological characteristics and immunohistochemical phenotypes of three cases of PAS, and aims to facilitate the identification of this lethal disease. In the present study, the data from clinical, histopathological and immunohistochemical examinations of three patients with PAS, whose diagnoses were confirmed by surgical biopsy conducted at the Beijing Anzhen Hospital (Beijing, China) between 2008 and 2012, were retrospectively analyzed. The patients (two women and one man; average age, 41.3 years old) presented with dyspnea on exertion. In addition, two of the patients experienced chest tightness, and one patient experienced intermittent syncope. Computed tomography pulmonary angiography revealed that two of the patients possessed a filling defect in the main, left and right pulmonary arteries, and one patient possessed a filling defect in the right upper pulmonary artery. Macroscopically, the PAS appeared as a mucoid intraluminal or nodular sessile mass spreading along the pulmonary artery. Microscopically, the tumor consisted of spindle cells with fascicular and storiform patterns, and was accompanied by necrosis and stromal myxoid changes. Immunohistochemically, vimentin, desmin and cluster of differentiation 34 were highly expressed in the patient that was diagnosed with intimal sarcoma, while vimentin and α-smooth muscle actin were highly expressed in the other two patients, who were diagnosed with leiomyosarcoma. PAS is often misdiagnosed due to nonspecific clinical manifestations and radiological features. Therefore, the diagnosis of PAS should be based on typical morphological features and immunohistochemical analysis of the tumor tissue. PMID:27073558

  17. Microscopic Analysis of Activated Sludge. Training Manual.

    ERIC Educational Resources Information Center

    Office of Water Program Operations (EPA), Cincinnati, OH. National Training and Operational Technology Center.

    This training manual presents material on the use of a compound microscope to analyze microscope communities, present in wastewater treatment processes, for operational control. Course topics include: sampling techniques, sample handling, laboratory analysis, identification of organisms, data interpretation, and use of the compound microscope.…

  18. An immunohistochemical study of Na+/I- symporter in human thyroid tissues and salivary gland tissues.

    PubMed

    Jhiang, S M; Cho, J Y; Ryu, K Y; DeYoung, B R; Smanik, P A; McGaughy, V R; Fischer, A H; Mazzaferri, E L

    1998-10-01

    The human Na+/I- symporter (hNIS) is the plasma membrane protein that mediates active iodide uptake into several tissues, such as the thyroid and salivary glands. To study the distribution and cellular localization of the hNIS protein, we have generated a polyclonal antibody that could detect the hNIS protein by immunohistochemical staining on tissue sections. In normal thyroids, hNIS expression is heterogeneous, and it is only detected in sporadic thyrocytes of a given follicle. The hNIS protein was not detected in thyroid carcinomas, yet it was detected in the majority of thyrocytes in Graves' thyroids. In salivary glands, hNIS protein was not detected in acinar cells, but it was detected in ductal cells. The hNIS proteins are clustered in the basal and lateral membranes in cells stained positive for hNIS.

  19. Immunohistochemical characteristics of epithelial cell rests of Malassez during cementum repair.

    PubMed

    Hasegawa, Naohiko; Kawaguchi, Hiroyuki; Ogawa, Tetsuji; Uchida, Takashi; Kurihara, Hidemi

    2003-02-01

    To clarify the roles of epithelial cell rests of Malassez (ECRM) during periodontal repair, experimental root resorption was induced in rats and then the ECRM that existed in periodontal ligament during cementum repair was investigated using morphological and immunohistochemical approaches. At day 7, after mechanical injury, root resorption was observed and ECRM were present adjacent to the site of resorption lacunae. They were observed in periodontal ligament adjacent to site of the resorption lacunae. These ECRM were immunoreactive for bone morphogenetic protein-2. During the stage of early cementum repair, the ECRM were immunoreactive for osteopontin and ameloblastin. They strongly reacted to proliferating cell nuclear antigen. In uninjured control sections, ECRM located in the periodontal ligament adjacent to cementum were not immunoreactive for any antibodies. These findings suggested that ECRM may be related to cementum repair by activating their potential to secrete matrix proteins which have been expressed in tooth development. PMID:12558937

  20. Gravity stress elevates the nociceptive threshold level with immunohistochemical changes in the rat brain

    NASA Astrophysics Data System (ADS)

    Kumei, Yasuhiro; Shimokawa, Reiko; Kimoto, Mari; Kawauchi, Yasuko; Shimokawa, Hitoyata; Makita, Koshi; Ohya, Keiichi; Toda, Kazuo

    2001-08-01

    Young Wistar male rats were exposed to 2G hypergravity by continuous centrifugation for 15 minutes. The nociceptive threshold was measured by using the von Frey type filament on the rat skin surfaces after hypergravity exposure. Following the hypergravity exposure, rats were sacrificed with anesthesia, then perfused and fixed for immunohistochemical examination. The 2G hypergravity elevated the nociceptive threshold up to 2-fold and induced analgesic effects on rats that remained for 2 hours after termination of centrifugation. Expression of Fos-immunoreactive proteins was prominently induced by 2G hypergravity in the arcuate nucleas and the paraventricular nucleus of the hypothalamus. The 15-minute flash exposure to 2G hypergravity induced pain suppression in rats, which might be attributed to change of neuronal activity in rat hypothalamus.

  1. Immunohistochemical and molecular biomarkers in Coris julis exposed to environmental contaminants.

    PubMed

    Fasulo, Salvatore; Mauceri, Angela; Maisano, Maria; Giannetto, Alessia; Parrino, Vincenzo; Gennuso, Florinda; D'Agata, Alessia

    2010-07-01

    When a contaminant interacts with biotic components of a marine ecosystem, it causes a series of changes that can compromise an entire community (Stebbing, 1985). This present study wants to focus on changes in the gills of a bioindicator benthic organism, Coris julis, collected in Milazzo (Messina, Italy), characterized by a strong anthropical impact), compared with individuals from the control site (Marinello, Messina). RT-PCR has been used for both MT and HSP70, and the respective mRNAs have been visualized by FISH. MT and HSP70 expression levels increased in individuals collected in Milazzo. The presence of numerous apoptotic and proliferating cells and the analysis of several neuronal markers by immunohistochemical method give information about the adaptation to a heavy metal mixture. The obtained results show that, in stressed fishes, defensive processes increase to maintain the normal functions of the organs more exposed to the action of polluted substances. PMID:20129668

  2. Intrapulmonary solitary fibrous tumor diagnosed by immunohistochemical and genetic approaches: report of a case.

    PubMed

    Sagawa, Motoyasu; Ueda, Yoshimichi; Matsubara, Fujitsugu; Sakuma, Hiroshi; Yoshimitsu, Yutaka; Aikawa, Hirokazu; Usuda, Katsuo; Minato, Hiroshi; Sakuma, Tsutomu

    2007-01-01

    Although solitary fibrous tumors (SFTs) of the pleura are not uncommon, intrapulmonary SFTs are extremely rare. A 72-year-old woman was admitted to our hospital for an investigation of an enlarging intrapulmonary tumor. Because a definitive diagnosis could not be readily established, a pulmonary wedge resection under video-assisted thoracic surgery was performed. Grossly, the tumor was white, well circumscribed, and separate from the pleural surface. Histologically, it consisted of spindle cells proliferating in a vague fascicular pattern, with many dilated capillaries, and intermingled glandular components. These findings suggested a differential diagnosis that included SFT and nonchondromatous pulmonary hamartoma. On immunohistochemical analysis, the spindle cells showed a strong positive reaction to the CD34 antigen. Interphase fluorescent in situ hybridization revealed an absence of HMGA-1 and -2 translocations. These results supported a diagnosis of SFT. A genetic approach may therefore be useful in the differentiation of SFT from nonchondromatous hamartoma.

  3. Spindle cell carcinoma of the mandible: Clinicopathological and immunohistochemical characteristics.

    PubMed

    Al-Bayaty, Haytham; Balkaran, Ramaa L

    2016-01-01

    Spindle cell carcinoma, a rare variant of squamous cell carcinoma, has propensity to occur in the upper aero digestive tract, including the oral mucosa. In this oral pathology communication, we report the occurrence of this neoplasm in the left mandible as a large fleshy growth with destruction of bone in a 73-year-old Afro-Trinidadian female. The distinction of this tumor from other malignant spindle cell mesenchymal tumors is important. Selective sampling of this specimen for possible transitional areas of squamous and spindle cell appearance, immunohistochemical staining for cytokeratin, vimentin, and S-100 protein are helpful in establishing the diagnosis. According to the patient's insistence, debulking of the tumor was performed under general anesthesia. Eight months later the patient succumbed to the disease.

  4. [Bone marrow involvement in ovarian cancer determined by immunohistochemical methods].

    PubMed

    Gabriel, M; Obrebowska, A; Spaczyński, M

    2000-01-01

    Atypical epithelial cells in the bone marrow of patients with ovarian cancer were evaluated using immunohistochemical techniques. We investigated cytospin preparations of bone marrow taken from 9 women with benign ovarian tumors and 59 women with malignant ovarian tumors. Two monoclonal antibodies (NCL-C11 and NCL-CA 125) were used. With both antibodies we were able to detect keratin and CA 125 antigen expression in the bone marrow of 9 (18.4%) of the patients with ovarian cancer. With regard to the wide histological differentiation of ovarian carcinomas, the presence of atypical epithelial cells in the bone marrow was required as a prognostic factor for survival and relapses. This should be investigated in a larger study group. PMID:11326158

  5. Classification and immunohistochemical scoring of breast tissue microarray spots.

    PubMed

    Amaral, Telmo; McKenna, Stephen J; Robertson, Katherine; Thompson, Alastair

    2013-10-01

    Tissue microarrays (TMAs) facilitate the survey of very large numbers of tumors. However, the manual assessment of stained TMA sections constitutes a bottleneck in the pathologist's work flow. This paper presents a computational pipeline for automatically classifying and scoring breast cancer TMA spots that have been subjected to nuclear immunostaining. Spots are classified based on a bag of visual words approach. Immunohistochemical scoring is performed by computing spot features reflecting the proportion of epithelial nuclei that are stained and the strength of that staining. These are then mapped onto an ordinal scale used by pathologists. Multilayer perceptron classifiers are compared with latent topic models and support vector machines for spot classification, and with Gaussian process ordinal regression and linear models for scoring. Intraobserver variation is also reported. The use of posterior entropy to identify uncertain cases is demonstrated. Evaluation is performed using TMA images stained for progesterone receptor.

  6. Parvovirus infection: an immunohistochemical study using fetal and placental tissue.

    PubMed

    Li, Jing Jing; Henwood, Tony; Van Hal, Sebastian; Charlton, Amanda

    2015-01-01

    Parvovirus B19 infection causes 5% to 15% of cases of nonimmune hydrops fetalis. The aim of our study was to evaluate the use of immunohistochemistry in diagnosing parvovirus infection in fetal and placental tissue during routine fetal and perinatal autopsies. Histology slides of 20 cases of confirmed parvovirus infection were reviewed, and immunohistochemistry was applied to selected blocks of fetal and placental tissue. Immunohistochemistry was positive in all 20 cases, and histologic viral inclusions were seen in 19 cases. Immunohistochemical staining was closely correlated with histology and was more sensitive than histology in detecting virally infected cells, especially in autolyzed tissue. All cases also had confirmatory evidence of parvovirus infection by polymerase chain reaction of fetal liver and positive maternal serology, where it was available. We conclude that parvovirus immunohistochemistry is a reliable method for diagnosing parvovirus infection, especially in autolyzed tissue where histologic assessment may be suboptimal.

  7. Immunohistochemical investigations of genital ulcers caused by Haemophilus ducreyi.

    PubMed

    Abeck, D; Freinkel, A L; Korting, H C; Szeimis, R M; Ballard, R C

    1997-09-01

    To gain information on the specific composition of the inflammatory infiltrate of genital ulcers caused by Haemophilus ducreyi, biopsies of 6 genital ulcers which were diagnosed as chancroid on clinical and microbiological grounds were subjected to immunohistochemical investigations after conventional haematoxylineosin staining. A variety of antibodies reactive against B- and T-cells, plasma cells and granulocytes were used with each tissue sections. The lymphocytic infiltrate of chancroid ulcers consisted of both B- and T-lymphocytes and showed a cluster-like formation. B-lymphocytes were preferentially localized perivascularly in the middle layer, T-lymphocytes mainly in the deep layer of the inflamed oedematous tissue. Results stress the importance of both B- and T-cell mediated immune responses in Haemophilus ducreyi infection.

  8. COTA (colon-ovarian tumor antigen). An immunohistochemical study.

    PubMed

    Pant, K D; Fenoglio-Preiser, C M; Berry, C O; Zamora, P O; Ram, M D; Fulks, R M; Rhodes, B A

    1986-07-01

    A goat anti-serum was prepared against mucinous ovarian cyst fluid and absorbed with normal colon and a variety of normal tissues until the only residual immunoreactivity was directed against colon cancer and ovarian tumor mucin. The set of antigenic determinants defined by this anti-serum has been called COTA, standing for colon-ovarian-tumor-antigen. This highly absorbed anti-serum (anti-COTA) was used for immunohistochemical staining of 42 different tissues in parallel with staining with a goat anti-CEA, which was also highly absorbed. The results suggest that COTA is a highly sensitive and specific antigen for colon carcinoma and may have potential for the early detection of malignant changes predictive of cancer of the colon.

  9. Carcinosarcoma of the submandibular salivary gland. Immunohistochemical findings.

    PubMed

    Bleiweiss, I J; Huvos, A G; Lara, J; Strong, E W

    1992-04-15

    Carcinosarcomas of the salivary glands are rare lesions that generally have been associated with benign mixed tumors. The authors report a case of a submandibular gland lesion, which occurred in a 64-year-old man, that was composed of intermingled ductal type adenocarcinoma and osteogenic sarcoma with a large component of osteoclast-like giant cells. The local recurrence of the tumor was entirely sarcomatous with no epithelial component observed. There was no histologic evidence of a preexisting or coexisting pleomorphic adenoma. Immunohistochemical studies confirmed two separate populations of tumor cells, corresponding to the histologic growth pattern. The authors review the literature and discuss histogenetic implications of distinction between de novo carcinosarcoma and carcinosarcoma in association with pleomorphic adenoma.

  10. Active vision in satellite scene analysis

    NASA Technical Reports Server (NTRS)

    Naillon, Martine

    1994-01-01

    In earth observation or planetary exploration it is necessary to have more and, more autonomous systems, able to adapt to unpredictable situations. This imposes the use, in artificial systems, of new concepts in cognition, based on the fact that perception should not be separated from recognition and decision making levels. This means that low level signal processing (perception level) should interact with symbolic and high level processing (decision level). This paper is going to describe the new concept of active vision, implemented in Distributed Artificial Intelligence by Dassault Aviation following a 'structuralist' principle. An application to spatial image interpretation is given, oriented toward flexible robotics.

  11. Merkel cell carcinoma with an unusual immunohistochemical profile.

    PubMed

    Pilloni, L; Manieli, C; Senes, G; Ribuffo, D; Faa, G

    2009-12-29

    The clinical and morphological picture of Merkel cell carcinoma (MCC) may be rather challenging; therefore, the immunohistochemical profile plays a relevant role in confirming the microscopic diagnosis. A panel of antibodies including cytokeratins 20, 7 and epithelial membrane antigen, and neuron-specific enolase is used in confirming the morphological diagnosis of MCC. The majority of MCCs express CK20 and are CK7-negative. Herein, we present a case of primary cutaneous neuroendocrine carcinoma with an atypical immunohistochemical pattern. A 83-years old woman presented with a painless plaque, red to violaceous in colour, located in the leg. The skin tumor was excided, formalin-fixed and paraffinembedded. Tissue sections were immunostained with a panel of antibodies routinely utilized in complex primary skin tumors for evidencing epithelial and neuroendocrine differentiation of tumor cells. The neuroendocrine differentiation of tumor cells was evidenced by their immunoreactivity for synaptophysin, chromograninA and neuron-specific enolase. Tumor cells also showed diffuse cytoplasmic staining for CK7. No immunoreactivity was detected for CK20 and thyroid transcription factor-1. Our data, together with previous rare reports of CK20-/CK7+ MCCs, lay stress on the importance of routinely utilizing a panel of antibodies in the differential diagnosis of complex primary carcinomas of the skin and may have important implications in expanding the differential diagnosis of skin tumors. In particular, caution should be taken in excluding the diagnosis of MCC only on the basis of the absence of reactivity of tumor cells for CK20, favouring the wrong diagnosis of less aggressive skin tumors. PMID:22073365

  12. Passive versus active mitigation cost analysis

    SciTech Connect

    Parazin, R.J.; Galbraith, J.D.

    1995-04-01

    The scope of this task is to assess the impact of mitigation alternatives for Tanks 241-SY-101 and 241-SY-103 on the Project W-236A Multi-Function Waste Tank Facility. This assessment and other related tasks are part of an Action Plan Path Forward prepared by the Tank Waste Remediation System (TWRS) Life Extension and Transition Program. Task 3.7 of the Action Plan for Project W-236A MWTF analyzed the comparative cost/risk of two hydrogen gas mitigation alternatives (active versus passive) to recommend the most appropriate course of action to resolve the hydrogen gas safety issue. The qualitative success of active mitigation has been demonstrated through Tank 241-SY-101 testing. Passive mitigation has not been demonstrated but will be validated by laboratory test work performed under Task 3.1 of the Action Plan. It is assumed for this assessment that the uncertainties associated with the performance of either alternative is comparable. Determining alternative specific performance measures beyond those noted are not in the scope of this effort.

  13. Activation analysis of the compact ignition tokamak

    SciTech Connect

    Selcow, E.C.

    1986-01-01

    The US fusion program has completed the conceptual design of a compact tokamak device that achieves ignition. The high neutron wall loadings associated with this compact deuterium-tritium-burning device indicate that radiation-related issues may be significant considerations in the overall system design. Sufficient shielding will be requied for the radiation protection of both reactor components and occupational personnel. A close-in igloo shield has been designed around the periphery of the tokamak structure to permit personnel access into the test cell after shutdown and limit the total activation of the test cell components. This paper describes the conceptual design of the igloo shield system and discusses the major neutronic concerns related to the design of the Compact Ignition Tokamak.

  14. Phosphorus Determination by Derivative Activation Analysis: A Multifaceted Radiochemical Application.

    ERIC Educational Resources Information Center

    Kleppinger, E. W.; And Others

    1984-01-01

    Although determination of phosphorus is important in biology, physiology, and environmental science, traditional gravimetric and colorimetric methods are cumbersome and lack the requisite sensitivity. Therefore, a derivative activation analysis method is suggested. Background information, procedures, and results are provided. (JN)

  15. Prompt Gamma-Ray Neutron Activation Analysis (PGNAA) for Elemental Analysis

    SciTech Connect

    Robin P. Gardner

    2006-04-11

    This research project was to improve the prompt gamma-ray neutron activation analysis (PGNAA) measurement approach for bulk analysis, oil well logging, and small sample thermal enutron bean applications.

  16. Analysis of body calcium (regional changes in body calcium by in vivo neutron activation analysis)

    NASA Technical Reports Server (NTRS)

    Suki, W.; Johnson, P. C.; Leblanc, A.; Evans, H. J.

    1981-01-01

    The effect of space flight on urine and fecal calcium loss was documented during the three long-term Skylab flights. Neutron activation analysis was used to determine regional calcium loss. Various designs for regional analysis were investigated.

  17. Analysis of supercooling activities of surfactants.

    PubMed

    Kuwabara, Chikako; Terauchi, Ryuji; Tochigi, Hiroshi; Takaoka, Hisao; Arakawa, Keita; Fujikawa, Seizo

    2014-08-01

    Supercooling-promoting activities (SCAs) of 25 kinds of surfactants including non-ionic, anionic, cationic and amphoteric types were examined in solutions (buffered Milli-Q water, BMQW) containing the ice nucleation bacterium (INB) Erwinia ananas, silver iodide (AgI) or BMQW alone, which unintentionally contained unidentified ice nucleators, by a droplet freezing assay. Most of the surfactants exhibited SCA in solutions containing AgI but not in solutions containing the INB E. ananas or BMQW alone. SCAs of many surfactants in solutions containing AgI were very high compared with those of previously reported supercooling-promoting substances. Cationic surfactants, hexadecyltrimethylammonium bromide (C16TAB) and hexadecyltrimethylammonium chloride (C16TAC), at concentrations of 0.01% (w/v) exhibited SCA of 11.8 °C, which is the highest SCA so far reported. These surfactants also showed high SCAs at very low concentrations in solutions containing AgI. C16TAB exhibited SCA of 5.7 °C at a concentration of 0.0005% (w/v). PMID:24792543

  18. Analysis of supercooling activities of surfactants.

    PubMed

    Kuwabara, Chikako; Terauchi, Ryuji; Tochigi, Hiroshi; Takaoka, Hisao; Arakawa, Keita; Fujikawa, Seizo

    2014-08-01

    Supercooling-promoting activities (SCAs) of 25 kinds of surfactants including non-ionic, anionic, cationic and amphoteric types were examined in solutions (buffered Milli-Q water, BMQW) containing the ice nucleation bacterium (INB) Erwinia ananas, silver iodide (AgI) or BMQW alone, which unintentionally contained unidentified ice nucleators, by a droplet freezing assay. Most of the surfactants exhibited SCA in solutions containing AgI but not in solutions containing the INB E. ananas or BMQW alone. SCAs of many surfactants in solutions containing AgI were very high compared with those of previously reported supercooling-promoting substances. Cationic surfactants, hexadecyltrimethylammonium bromide (C16TAB) and hexadecyltrimethylammonium chloride (C16TAC), at concentrations of 0.01% (w/v) exhibited SCA of 11.8 °C, which is the highest SCA so far reported. These surfactants also showed high SCAs at very low concentrations in solutions containing AgI. C16TAB exhibited SCA of 5.7 °C at a concentration of 0.0005% (w/v).

  19. Overview af MSFC's Applied Fluid Dynamics Analysis Group Activities

    NASA Technical Reports Server (NTRS)

    Garcia, Roberto; Griffin, Lisa; Williams, Robert

    2004-01-01

    This paper presents viewgraphs on NASA Marshall Space Flight Center's Applied Fluid Dynamics Analysis Group Activities. The topics include: 1) Status of programs at MSFC; 2) Fluid Mechanics at MSFC; 3) Relevant Fluid Dynamics Activities at MSFC; and 4) Shuttle Return to Flight.

  20. Spatiotemporal Data Mining, Analysis, and Visualization of Human Activity Data

    ERIC Educational Resources Information Center

    Li, Xun

    2012-01-01

    This dissertation addresses the research challenge of developing efficient new methods for discovering useful patterns and knowledge in large volumes of electronically collected spatiotemporal activity data. I propose to analyze three types of such spatiotemporal activity data in a methodological framework that integrates spatial analysis, data…

  1. Understanding Tensions: Activity Systems Analysis of Cross-Continental Collaboration

    ERIC Educational Resources Information Center

    Ryder, LanHui Zhang; Yamagata-Lynch, Lisa

    2014-01-01

    Using the lens of Vygotsky's sociocultural theory, activity theory, and Engeström's activity systems analysis, this qualitative study explores students' experiences in the context of a sixteen-week transpacific collaboration between seven students at Northern Illinois University (NIU) and seven students from Shandong Normal…

  2. Project-Based Language Learning: An Activity Theory Analysis

    ERIC Educational Resources Information Center

    Gibbes, Marina; Carson, Lorna

    2014-01-01

    This paper reports on an investigation of project-based language learning (PBLL) in a university language programme. Learner reflections of project work were analysed through Activity Theory, where tool-mediated activity is understood as the central unit of analysis for human interaction. Data were categorised according to the components of human…

  3. Pediatric intracranial ependymomas: prognostic relevance of histological, immunohistochemical, and flow cytometric factors.

    PubMed

    Zamecnik, Josef; Snuderl, Matija; Eckschlager, Tomas; Chanova, Marketa; Hladikova, Marie; Tichy, Michal; Kodet, Roman

    2003-10-01

    The correlation between the histological features and clinical outcome remains poor in pediatric intracranial ependymomas. We performed a retrospective study of a group of 31 patients (diagnosed from 1985 to 1995) to assess prognostic implications of the current grading system, of histological and immunohistochemical features, and of ploidy status estimated by flow cytometry. Immunoexpression of a broad spectrum of antigens was evaluated, including MIB-1, topoisomerase-IIalpha, cyclin D1, glial and epithelial proteins (GFAP, EMA, cytokeratins), molecules involved in controlling apoptosis (bcl-2, caspase-3/CPP32), and p53 oncoprotein. Univariate and multivariate statistical analyses were performed to evaluate the influence of each variable on both the progression free survival (PFS) and the overall survival (OS) with at least 7-year follow up. Although we showed a significant correlation between histological grade and prognosis, the current grading system failed in predicting outcome in nearly one third of individual cases. Problems with interpathologist reproducibility were also demonstrated. The extent of surgical resection was the only clinical factor that was associated with survival. Both the PFS and the OS were significantly decreased for the following pathological variables: increased cellularity (>300 nuclei per HPF), mitotic activity of >7 per 10 HPF, increased MIB-1 labeling index (LI), topoisomerase-IIalpha LI, S-phase fraction, and p53 and bcl-2 positivity. Increased cyclin D1 LI was demonstrated to have only a marginally significant impact on PFS. A flow chart modeling was further performed to formulate a scheme for discriminating of prognostic subgroups. Based on that, p53 immunopositivity and/or MIB-1 LI of >5% (after subtotal resection) or MIB-1 LI of >15% (after complete resection) were the strongest indicators of the tumor's aggressive behavior and of a poor prognosis of the disease. Foci of hypercellularity should be specifically looked for in

  4. EGFR and gastrointestinal stromal tumor: an immunohistochemical and FISH study of 82 cases.

    PubMed

    Lopes, Lisandro F; Bacchi, Carlos E

    2007-09-01

    Gastrointestinal stromal tumor is the most common mesenchymal neoplasm of the gastrointestinal tract. Mutually exclusive KIT or platelet-derived growth factor receptor-alpha mutations are key events in gastrointestinal stromal tumor pathogenesis, and specific treatment targeting KIT/platelet-derived growth factor receptor-alpha activation is available. Epidermal growth factor receptor plays an important role in cancer biology and also constitutes a promising molecular target of therapy. Very few reports have been published in the literature about the relationship between gastrointestinal stromal tumor and epidermal growth factor receptor. The aim of this study was to investigate epidermal growth factor receptor immunohistochemical expression and epidermal growth factor receptor gene amplification in 82 consecutive gastrointestinal stromal tumor cases using tissue microarray technique. Hematoxylin- and eosin-stained sections and clinical information were reviewed, and expression of CD117 (KIT), CD34 and epidermal growth factor receptor was investigated by immunohistochemistry. Epidermal growth factor receptor gene copy number was determined using fluorescence in situ hybridization. Immunohistochemistry revealed that CD117 and CD34 were expressed in 96 and 57% of tumors, respectively. Variable epidermal growth factor receptor protein immunohistochemical overexpression was detected in 96% of gastrointestinal stromal tumor cases, but none of the 75 cases with represented tumor tissue cores and countable fluorescence signals exhibited epidermal growth factor receptor gene amplification by fluorescence in situ hybridization. These results show that there is no correlation between epidermal growth factor receptor protein overexpression by immunohistochemistry and epidermal growth factor receptor gene amplification by fluorescence in situ hybridization. Considering that the mechanisms of epidermal growth factor receptor protein overexpression are not well understood and

  5. Nipple eczema, an indicative manifestation of atopic dermatitis? A clinical, histological, and immunohistochemical study.

    PubMed

    Song, Hyo Sang; Jung, Soo-Eun; Kim, You Chan; Lee, Eun-So

    2015-04-01

    Nipple eczema exhibits as a minor manifestation of atopic dermatitis (AD) or occurs as a single skin symptom on the nipple. To characterize the relationship between nipple eczema and AD, a clinical evaluation and an immunohistochemical study were performed. All cases of nipple eczema were confirmed histopathologically. We divided the patients with nipple eczema into 2 groups, namely, those with AD and those without AD, and compared several clinical features. Upon histological examination, the degree of inflammation was subjectively graded as mild, moderate, or severe by 2 separate investigators. Immunohistochemical stainings were performed by using antiinterleukin (IL)-4, anti-IL-13, anti-CD4, and anti-CD8 antibodies, and the results were scored semiquantitatively. In 43 cases evaluated, 12 were nipple eczema with AD. The clinical analysis and histological examination showed no significant differences between the groups. There were consistent findings of IL-4 expressions throughout the epidermis and IL-13 expression mainly in the perivascular area of the dermis. Although CD4 and CD8 were expressed in the cells in the dermis, CD8 expression was detected in the serocrusts of the epidermis. Expression levels of IL-4, IL-13, CD4, and CD8 exhibited no significant differences between the nipple eczema group with AD and the nipple eczema group without AD. Although nipple eczema may accompany AD, we found no definite differences in the degree or pattern of inflammation and cytokine expression level regardless of whether AD was present or not. Serocrust formation seemed to be mainly a collection of CD8-positive cells.

  6. Spatial heterogeneity analysis of brain activation in fMRI

    PubMed Central

    Gupta, Lalit; Besseling, René M.H.; Overvliet, Geke M.; Hofman, Paul A.M.; de Louw, Anton; Vaessen, Maarten J.; Aldenkamp, Albert P.; Ulman, Shrutin; Jansen, Jacobus F.A.; Backes, Walter H.

    2014-01-01

    In many brain diseases it can be qualitatively observed that spatial patterns in blood oxygenation level dependent (BOLD) activation maps appear more (diffusively) distributed than in healthy controls. However, measures that can quantitatively characterize this spatial distributiveness in individual subjects are lacking. In this study, we propose a number of spatial heterogeneity measures to characterize brain activation maps. The proposed methods focus on different aspects of heterogeneity, including the shape (compactness), complexity in the distribution of activated regions (fractal dimension and co-occurrence matrix), and gappiness between activated regions (lacunarity). To this end, functional MRI derived activation maps of a language and a motor task were obtained in language impaired children with (Rolandic) epilepsy and compared to age-matched healthy controls. Group analysis of the activation maps revealed no significant differences between patients and controls for both tasks. However, for the language task the activation maps in patients appeared more heterogeneous than in controls. Lacunarity was the best measure to discriminate activation patterns of patients from controls (sensitivity 74%, specificity 70%) and illustrates the increased irregularity of gaps between activated regions in patients. The combination of heterogeneity measures and a support vector machine approach yielded further increase in sensitivity and specificity to 78% and 80%, respectively. This illustrates that activation distributions in impaired brains can be complex and more heterogeneous than in normal brains and cannot be captured fully by a single quantity. In conclusion, heterogeneity analysis has potential to robustly characterize the increased distributiveness of brain activation in individual patients. PMID:25161893

  7. Neutron activation analysis for antimetabolites. [in food samples

    NASA Technical Reports Server (NTRS)

    1973-01-01

    Determination of metal ion contaminants in food samples is studied. A weighed quantity of each sample was digested in a concentrated mixture of nitric, hydrochloric and perchloric acids to affect complete solution of the food products. The samples were diluted with water and the pH adjusted according to the specific analysis performed. The samples were analyzed by neutron activation analysis, polarography, and atomic absorption spectrophotometry. The solid food samples were also analyzed by neutron activation analysis for increased sensitivity and lower levels of detectability. The results are presented in tabular form.

  8. Immunohistochemical study and mRNA cytokine profile of the local immune response in cattle naturally infected with Calicophoron daubneyi.

    PubMed

    Fuertes, Miguel; Manga-González, Yolanda; Benavides, Julio; González-Lanza, M Camino; Giráldez, Francisco Javier; Mezo, Mercedes; González-Warleta, Marta; Fernández, Miguel; Regidor-Cerrillo, Javier; Castaño, Pablo; Royo, Marcos; Ortega-Mora, Luis M; Pérez, Valentín; Ferreras, M Carmen

    2015-11-30

    In order to recognize the local immune response of the definitive host to Calicophoron daubneyi natural infection, an immunohistochemical study was carried out in the reticulum and rumen in 49 naturally infected cattle. The role of cytokines (IL-4 and IL-10 interleukins and IFN-γ) in the activation of specific defence mechanisms was evaluated by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) assays to study cytokine mRNA expression. In all infected animals, CD3+ T lymphocytes seemed to be the main element of the inflammatory infiltrate in the reticular and ruminal lamina propria at the point of the parasite adhesion. Intraepithelial globule leukocytes also showed immunolabelling for CD3. Most CD3+ cells also expressed CD4 (T cell helper) antigen although sporadic CD8+-cytotoxic lymphocytes were observed. Local expression of IFN-γ was observed in damaged papillae at the site of parasite attachment and in scattered cells in the lamina propria. B cells (CD79αcy+, CD45+ and IgG+) were found constantly in relation to lymphoid aggregates. MAC387 was expressed in squamous epithelium and in macrophages of the lamina propria of affected papillae. Macrophages in this location also stained positively for CD163 and CD68. Intraepithelial Langerhans cells and macrophages located in the lamina propria showed immunopositivity for MHCII in the affected areas. RT-qPCR analysis confirmed a statistical significant increase of IFN-γ, and IL-10 expression (p<0.01) in the rumen associated with the presence of flukes. These findings suggest a predominant Th1 polarized local immune response with the probable involvement of Th regulatory cells in cattle C. daubneyi natural infection. PMID:26508417

  9. Cutaneous pseudolymphoma: a case report with an immunohistochemical study

    PubMed Central

    Terada, Tadashi

    2013-01-01

    Cutaneous pseudolymphoma (C-PSL) is defined as reactive polyclonal benign lymphoproliferative process predominantly composed of either B-cells or T-cells, localized or disseminated. It heals spontaneously after cessation of the causative factor (e.g. drugs) or after non-aggressive treatment. The author herein presents a case of C-PSL with an immunohistochemical study. A 78-year-old man consulted our hospital because of slightly itching skin swelling on the arm. He denied insect bite and traumatic injury. His usual intake drugs were drugs of hypertension, hyperlipidemia, diabetes mellitus, and emotional disorders. Physical examination showed mildly erosive swelling of the am. The lesion measured 1 x 1 x 0.2 cm. Biopsy of the lesion was taken, and it revealed excessive proliferation of small lymphoid cells. The lymphoid cells lacked apparent atypical features and appeared matures. Lymphoblastic cells with nucleoli were scattered. Nodular structures were also seen in the lower dermis. Immunohistochemically, the lymphoid cells were positive for vimentin, CD3, CD4, CD5, CD8, CD10, CD15, CD20, CD23, CD30, CD43, CD38, CD138, CD45RO, CD79α, bcl-2, bcl-6, κ-chain, λ-chain, and Ki-67 (labeling index=7%). No light chain restriction is seen. The lymphoblastic cells were positively labeled for CD15 and CD30. Plasma cells positive for CD38, CK79α and CD138 were seen in a significant amounts. They were negative for cytokeratin (CK) CAM5.2, CKAE1/3, CK34BE12, CK5/6, CK7, CK8, CK18, CK19, CK20, EMA, CEA, CD56, CD57, p53, KIT, PDGFRA, and cyclin D1. Because the constituent cells were both B-cells including plasm cells and T-cells, no light-chain restriction was seen, and no histological atypia was seen, a diagnosis of cutaneous pseudolymphoma was made. The low Ki-67 labeling and negative p53 also suggested the diagnosis. The lesion slightly reduced in size (from 1 cm to 0.7 cm), the causative agent was still unknown 11 months after the biopsy. PMID:23638232

  10. Lubricin expression in human osteoarthritic knee meniscus and synovial fluid: a morphological, immunohistochemical and biochemical study.

    PubMed

    Musumeci, Giuseppe; Trovato, Francesca Maria; Loreto, Carla; Leonardi, Rosalia; Szychlinska, Marta Anna; Castorina, Sergio; Mobasheri, Ali

    2014-06-01

    The purpose of this study was to investigate the expression of lubricin, the product of the human PRG4 (proteoglycan 4) gene, in menisci and synovial fluid from normal donors and patients with osteoarthritis (OA), using a combination of histology, immunohistochemistry, ELISA and Western blotting analysis, to provide further insight on the role of this protein in the progression of OA and pathological processes in the meniscus. Lubricin expression was studied in samples from 40 patients and in 9 normal donors after arthroscopic partial meniscectomy. Histological analysis confirmed normal microanatomy and the absence of structural changes in control samples. Menisci derived from OA patients showed evidence of structural alterations, fibrillations and clefts. Immunohistochemical analysis revealed very strong lubricin immunostaining in normal menisci in contrast to weak/moderate staining seen in osteoarthritic menisci. Quantitative ELISA and Western blot analysis confirmed the above results. The findings of this study support the notion that changes in lubricin expression and boundary-lubricating ability of cartilage is followed by the development of OA. This study could provide the biological foundation for the development of novel therapeutic treatments, to be applied before the surgery, for the prevention of post-traumatic cartilage damage.

  11. Immunohistochemical localization of Nox in mouse circumvallate papillae.

    PubMed

    Kashiwabara, Yoshiaki; Ambe, Kimiharu; Nakagawa, Toshihiro; Watanabe, Hiroki

    2015-12-01

    Recently it has been reported that reactive oxygen species plays an important role in several physiological processes. Reactive oxygen species are generated by reactive oxygen-synthesizing enzymes (Nox). We immunohistochemically investigated expression and localization of the Nox family in a mouse circumvallate papillae. In the epithelium of the circumvallate papilla, Nox 1, 2, 3, and 4, Noxo1, and Noxa1 were expressed. In the circumvallate papilla, Nox2 was more weakly expressed in the lateral than in the upper part, and Nox3 was not expressed. In the taste buds, Nox 1, 2, 3, and 4, Noxo1, and Noxa1 were expressed; the Nox expression pattern varied with the cell type. In type II cells, Nox 1, 3, and 4, Noxo1, and Noxa1 were expressed. In type III cells, Nox2, besides Nox 1, 3, and 4, Noxo1, and Noxa1, were specifically expressed, unlike in other taste bud cells. In the mouse circumvallate papilla, Nox is always expressed, suggesting the generation of reactive oxygen species. Of note, all cells comprising taste buds expressed Nox, with each showing a specific Nox expression pattern.

  12. Immunohistochemical demonstration of lumbar intervertebral disc innervation in the dog.

    PubMed

    Willenegger, S; Friess, A E; Lang, J; Stoffel, M H

    2005-04-01

    Low back pain is a common ailment in dogs, particularly in specific breeds such as the German shepherd dog. A number of structures such as facet joint capsules, ligaments, dorsal root ganglia, periosteum, vertebral endplates and meninges have been associated with this condition. Yet, in spite of all diagnostic efforts, the origin of pain remains obscure in a substantial proportion of all cases. A further structure often being involved in vertebral column disorders is the intervertebral disc. The presence of nerves, however, is a precondition for pain sensation and, consequently, structures lacking innervation can be left out of consideration as a cause for low back pain. Nerve fibres have been demonstrated at the periphery of the intervertebral disc in man, rabbit and rat. With regard to the dog, however, the extent of intervertebral disc innervation is still being disputed. The goal of the present study, therefore, was to substantiate and expand current knowledge of intervertebral disc innervation. Protein gene product (PGP) 9.5 was used for immunohistochemical examination of serial transversal and sagittal paraffin sections of lumbar discs from adult dogs. This general marker revealed nerve fibres to be confined to the periphery of the intervertebral discs. These results indicate that even limited pathological processes affecting the outer layers of the intervertebral disc are prone to cause low back pain.

  13. Immunohistochemical demonstration of vitamin D receptor distribution in goat intestines.

    PubMed

    Boos, Alois; Riner, Katharina; Hässig, Michael; Liesegang, Annette

    2007-01-01

    Vitamin D (VD) plays an important role in calcium homeostasis. 1,25-Dihydroxycholecalciferol or calcitriol modulates gene transcription via nuclear VD receptors (VDR). In the intestines, VD promotes calcium resorption via VDR. VDR has not been systematically assessed within the intestine in any species. We therefore present a semiquantitative immunohistochemical study of the distribution patterns of VDR in goat intestines. Intestinal tissue probes were collected from 5 lambs and 5 non-lactating non-pregnant dams, fixed in formalin, embedded in paraffin and assessed for VDR. Nuclear VDR immunoreaction was scored semiquantitatively. VDR exhibited a segment-specific distribution pattern. Goblet cells were always devoid of VDR. Enterocytes within the surface epithelium and the superficial crypts generally demonstrated only a weak immunoreaction along the length of the intestine, while basally and/or intermediately located crypt epithelial cells exhibited strong VDR immunoreactions in the duodenum, jejunum and colon descendens. The difference in VDR staining between deep and superficial locations was most prominent in the duodenum and less evident in the jejunum, ileum and colon descendens. Results demonstrate that VDR distribution exhibits cell type-, segment- and location-specific patterns in the goat. Data may serve as a basis for future experiments on the role of VDR in Ca metabolism.

  14. Immunohistochemical localization of endothelial cell markers in solitary fibrous tumor.

    PubMed

    Sawada, Namie; Ishiwata, Toshiyuki; Naito, Zenya; Maeda, Shotaro; Sugisaki, Yuichi; Asano, Goro

    2002-12-01

    Solitary fibrous tumor (SFT) is an uncommon tumor first reported in the pleura, but recently described in other tissues. CD34, which is expressed in hematopoietic stem cells, endothelial progenitor cells and vascular endothelial cells, is observed in most SFT and some investigators believe that its expression is a definitive marker of this tumor. In the present study, the expression of vascular endothelial cell markers, such as vascular endothelial growth factor receptor (VEGFR)-1 (flt-1), VEGFR-2 (flk-1/KDR), Tie-2 and c-Met, was examined in SFT to clarify the relationship between SFT and endothelial cells. By immunohistochemical staining of tumor cells from 26 patients, VEGFR-1 was detected in 24 (92%), VEGFR-2 in five (19%), Tie-2 in 14 (54%), and c-Met, a specific receptor of hepatocyte growth factor (HGF) in 23 patients (88%). Furthermore, VEGFR-3 (flt-4) immunoreactivity was detected in eight of 26 patients (31%). In contrast, VEGF, VEGF-C and HGF, which are ligands for the receptors, were not localized in the SFT cells. These findings indicate that most SFT may closely relate to vascular or lymphatic endothelial cells and the endothelial growth factors may contribute to the growth of SFT in a paracrine manner.

  15. Immunohistochemical investigation of wound healing in response to fractional photothermolysis

    NASA Astrophysics Data System (ADS)

    Helbig, Doris; Bodendorf, Marc Oliver; Grunewald, Sonja; Kendler, Michael; Simon, Jan C.; Paasch, Uwe

    2009-11-01

    Despite growing clinical evidence of ablative fractional photothermolysis (AFP), little is known about the spatiotemporal molecular changes within the targeted compartments. Six subjects received three different single AFP treatments using a scanned 250 μm CO2-laser beam. Spatiotemporal changes of skin regeneration were estimated by immunohistochemical investigation (HSP70, HSP72, HSP47, TGFβ, procollagen III, CD3, CD20, and CD68) in skin samples 1 h, 3 days, and 14 days postintervention. The remodeling was uniformly started by regrowth of the epidermal compartment followed by partial to complete replacement of the microscopic ablation zones (MAZ) by newly synthesized condensed procollagen III. From day 3 to 14, the number of macrophages as well as giant cells surrounding the MAZ increased. TGFβ expression was highest 1 h to 3 days following AFP. HSP70 and HSP72 expressions were highest 3-14 days postintervention in the spinocellular layer leading to an upregulation of HSP47. AFP performed by a scanned CO2-laser results in an early epidermal remodeling, which is followed by a dermal remodeling leading to a replacement of the MAZ with newly synthesized (pro)-collagen. During this, an inflammatory infiltrate with CD3+ and CD20+ cells surrounds the MAZ. The count of macrophages and giant cells involved in the replacement of the necrotic zones seems to be crucial for wound healing.

  16. Simple cysts of testis with immunohistochemical evidence of mesothelial origin.

    PubMed

    Kuwamoto, Satoshi; Okuno, Keisuke; Kato, Masako; Hayashi, Kazuhiko

    2009-09-01

    Simple cysts of the testis are an uncommon lesion found in adults as well as infants. Only a few histological and immunohistochemical studies for this lesion have been reported because the lesion have rarely undergone surgical excision or histological confirmation. Herein is reported an autopsy case of simple cysts of the right testis found incidentally in an 84-year-old man who died of hepatocellular carcinoma. The cysts were multilocular, 12 x 8 mm in size, separated from the tunica albuginea and filled with transparent and colorless serous fluid. Histology and immunohistochemistry showed that the cystic lesion had flattened or cuboidal single-layered lining cells that were positive for cytokeratin AE1/AE3, calretinin, D2-40, HBME-1, mesothelin and thrombomodulin, indicating its mesothelial origin. To the authors' knowledge this is the first English-language report describing the origin of simple cysts of the testis. Although the exact pathogenesis of simple cysts of the testis is as yet unknown and there seems to be more than one etiology, at least in the present case the cysts were confirmed to correspond to mesothelial inclusions.

  17. Delayed postburn blisters: an immunohistochemical and ultrastructural study.

    PubMed

    Bergman, R; David, R; Ramon, Y; Ramon, M; Kerner, H; Kilim, S; Peled, I; Friedman-Birnbaum, R

    1997-08-01

    This study was performed in an attempt to further elucidate the pathogenesis of delayed postburn blistering. Two cases were studied ultrastructurally and immunohistochemically, 1 with blisters on the recipient site of autologous split-thickness skin grafts and the other on the donor site. Ultrastructurally, the basement membrane was on the roof of the blisters in both cases, except for a single small blister in the first case where it was on the dermal floor. In the blister roofs, the basement membrane showed small or marked segments of discontinuity. In the adjacent non-blistered healed skin, the basement membrane was usually continuous, and anchoring fibrils were present. Immunoperoxidase staining on frozen sections, using antibodies to laminin, laminin 5, collagen IV, and collagen VII, showed a mostly continuous linear pattern in the adjacent non-blistered skin, which often became discontinuous near the blisters and markedly discontinuous in the blister roofs. In the blister floors, weakly stained linear or granular deposits of some of these components were sometimes also present. The results of this study support discontinuity of the basement membrane as the main anomaly in delayed postburn blistering. Disturbance in the reassembly or local breakdown of the basement membrane components might be the underlying defect.

  18. Ulcerative Granular Cell Tumor: A Clinicopathological and Immunohistochemical Study

    PubMed Central

    El-Khalawany, Mohamed; Mosbeh, Al-Sadat; Abd-Al Salam, Fatma; Abou-Bakr, Amany

    2011-01-01

    Granular cell tumor (GCT) is uncommonly presented with cutaneous ulcer. We examined the clinicopathological and immunohistochemical features of this ulcerative form in fourteen cases that may raise the awareness of this variant. The study included 11 males and 3 females with a mean age 31.5 ± 7.42 years. All cases were presented with large solitary ulcer with indurated base, elevated border, skin colored margin, and necrotic floor. Twelve lesions were located on the extremities and two lesions on the genital region. Histologically, the lesions showed dermal infiltrate composed of large polygonal cells with granular cytoplasm and characteristic infiltration of the dermal muscles in all cases. Immunostaining showed positive reaction for S100 (14/14), NSE (14/14), CD68 (5/14), and Vimentin (7/14) while HMB45, CK, EMA, and Desmin were negative. We hope that this paper increases the awareness of ulcerative GCT and consider it in the differential diagnosis of ulcerative lesions. PMID:22132340

  19. Immunohistochemical and ultrastructural detection of intestinal spirochetes in Thoroughbred horses.

    PubMed

    Shibahara, Tomoyuki; Kuwano, Atsutoshi; Ueno, Takanori; Katayama, Yoshinari; Ohya, Tatsuo; Taharaguchi, Sadao; Yamamoto, Shinji; Umemura, Takashi; Ishikawa, Yoshiharu; Kadota, Koichi

    2005-03-01

    Studies of equine intestinal spirochetes have long focused on intestinal contents alone, but intestinal spirochetosis has been reported recently in a 21-month-old Thoroughbred colt in Japan. To define the clinical and pathological significances of intestinal spirochetosis in several horses, an epizootiologic survey with histologic, immunohistochemical, and ultrastructural methods was conducted for Brachyspira antigen-containing intestinal spirochetes in 12 diseased or injured Thoroughbred horses, aged from 35 days to 17 years. Brachyspira antigen-containing spirochetes were found in 7 of 12 horses (58.3%) and were more frequent in the cecum than in other parts of the bowel. It was not clear whether the infection was clinically related to diarrhea or dysentery, but histopathology revealed a close association between the bacterial infection and epithelial hyperplasia. Crypt epithelium consisted mainly of goblet cells and showed frequent mitosis throughout its length. Inflammatory cells and congestion were also present. There were numerous spirochetes in the crypts, and some invaded the cecal and colonic epithelia and underlying lamina propria. Ultrastructurally, the spirochetes were divided into 4 types. Three types were identified in degenerative epithelial cells or intracellularly. Brachyspira antigen-containing intestinal spirochetes invading the mucosa were capable of causing epithelial hyperplasia in the cecum and colon in the horses. The findings in this study will increase awareness of the importance of intestinal spirochetosis and may also be helpful for diagnosis and treatment of this condition. PMID:15825495

  20. Hidradenoma Papilliferum With Oncocytic Metaplasia: A Histopathological and Immunohistochemical Study.

    PubMed

    Elbendary, Amira; Cochran, Eric; Xie, Qiang; Kabigting, Filamer; Pereira, Leanne; Elston, Dirk M; Heilman, Edward

    2016-06-01

    Hidradenoma papilliferum is a benign cutaneous adnexal neoplasm, commonly occurring in the vulva and perianal region of adult women. It has characteristic histopathological features composed of anastomosing and branching tubules, lined by columnar cells, and a basal layer of myoepithelial cells. A 39-year-old woman was evaluated for 2 asymptomatic labial masses. The histopathological examination revealed a Bartholin's cyst and a hidradenoma papilliferum. The latter contains a distinct area of oncocytic/oxyphilic metaplasia. Immunohistochemical stains revealed positive staining for gross cystic disease fluid protein (GCDFP)-15 and androgen receptor. GATA-3, a protein expressed in sweat glands, highlights a similar positive staining pattern with weaker staining in areas of oncocytic metaplasia. P63 highlighted the myoepithelial differentiation. In situ hybridization for Human Papilloma Virus 6, 11, 16, and 18 was negative. P53 was negative and Ki-67 was low, confirming its benign nature. Oncocytes are enlarged epithelial cells with voluminous eosinophilic granular cytoplasm resulting from staining of nonribosomal cytoplasmic components. Few reports documented it in hidradenoma papilliferum. Our case demonstrated a florid distinct appearance of this metaplasia. The immunoprofiles of this oncocytic metaplasia such as p53 negativity and positivity for androgen receptor and GCDFP-15 demonstrates similarity to apocrine metaplasia in the breast. The authors' case demonstrates the benign nature of oncocytic metaplasia and supports the common origin of oncocytic cells and columnar cells in hidradenoma papilliferum. PMID:27097337

  1. Immunohistochemical delineation of the conduction system. I: The sinoatrial node.

    PubMed

    Oosthoek, P W; Virágh, S; Mayen, A E; van Kempen, M J; Lamers, W H; Moorman, A F

    1993-09-01

    We have raised a mouse monoclonal antibody that reacts specifically with the myocytes of the sinoatrial node of the bovine heart. By use of this antibody (445-6E10) and antibodies against the gap junction protein connexin43, the periphery of the sinoatrial node and the distribution of gap junctions in the nodal region were studied. The reaction patterns of 445-6E10 and anti-connexin43 are exactly complementary; ie, connexin43 was not detected in the nodal myocytes but was clearly present in the atrial myocytes. Both reaction patterns demonstrate that nodal myocytes and atrial myocytes can unambiguously be distinguished by their characteristic molecular phenotype. The transitional nodal myocytes at the periphery of the node that have intermediate morphological and electrophysiological characteristics could now clearly be defined as nodal by our immunohistochemical criteria. The center of the node is surrounded by a region of interdigitating nodal and atrial bundles. Nodal bundles, coming from the center of the node, penetrate the atrial myocardium aligned at atrial bundles, forming histological connections between nodal and atrial myocytes at regular distances. This interdigitating arrangement of bundles of connexin43-negative nodal and connexin43-positive atrial myocytes is also found in the human and rat heart. We hypothesize that the architecture of the periphery of the node is important to prevent silencing of the pacemaking nodal myocytes by the atrium while ensuring a sufficient source loading of the nodal myocytes. PMID:8394223

  2. Postmortem Diagnosis of Anorexia Nervosa: An Endocrinological and Immunohistochemical Approach.

    PubMed

    Ishigami, Akiko; Tokunaga, Itsuo; Nishimura, Akiyoshi; Kubo, Shin-Ichi

    2016-01-01

    A female in her 30s was found dead after a fire. She was severely emaciated and had been diagnosed with anorexia nervosa (AN) about 5 years ago, but had not been treated recently. Therefore, we investigated not only her cause of death but also her condition of AN. Some of her organs weighed less than normal although no clear lesions were observed. In the pituitary gland, the number of follicle-stimulating hormone-immunopositive cells was markedly decreased although a normal number of thyroid-stimulating hormone-positive cells were detected. A histological examination of the ovary suggested that she had been suffering from amenorrhea. The thyroid gland was atrophic, and marked variations in follicle size were observed. Because we could not obtain enough volume of her blood for endocrinological examinations, we tried to investigate her endocrinological condition by immunohistochemistry. Immunohistochemical staining detected decreased triiodothyronine immunoreactivity and normal thyroxine immunoreactivity. The adrenal glands were also atrophic. Based on these findings, it was considered that she had been suffering from AN at the time of her death. The autopsy and other findings revealed that she had died of burning with carbon monoxide intoxication. J. Med. Invest. 63: 305-309, August, 2016. PMID:27644577

  3. Immunohistochemical vascular factor expression in canine inflammatory mammary carcinoma.

    PubMed

    Camacho, L; Peña, L; Gil, A González; Martín-Ruiz, A; Dunner, S; Illera, J C

    2014-07-01

    Human inflammatory breast carcinoma (IBC) and canine inflammatory mammary carcinoma (IMC) are considered the most malignant types of breast cancer. IMC has similar characteristics to IBC; hence, IMC has been suggested as a model to study the human disease. To compare the angiogenic and angioinvasive features of IMC with non-IMC, 3 canine mammary tumor xenograft models in female SCID mice were developed: IMC, comedocarcinoma, and osteosarcoma. Histopathological and immunohistochemical characterization of both primary canine tumors and xenografts using cellular markers pancytokeratin, cytokeratin 14, vimentin, and α-smooth muscle actin and vascular factors (VEGF-A, VEGF-D, VEGFR-3, and COX-2) was performed. Tumor cell proliferation index was measured by the Ki-67 marker. The xenograft models reproduced histological features found in the primary canine tumor and preserved the original immunophenotype. IMC xenografts showed a high invasive character with tumor emboli in the dermis, edema, and occasional observations of ulceration. In addition, compared with osteosarcoma and comedocarcinoma, the IMC model showed the highest vascular factor expression associated with a high proliferation index. Likewise, IMC xenografts showed higher COX-2 expression associated with VEGF-D and VEGFR-3, as well as a higher presence of dermal lymphatic tumor emboli, suggesting COX-2 participation in IMC lymphangiogenesis. These results provide additional evidence to consider vascular factors, their receptors, and COX-2 as therapeutic targets for IBC.

  4. Male breast carcinoma: a clinicopathological and immunohistochemical characterization study.

    PubMed

    Zhou, Ruoji; Yu, Lin; Zhou, Shuling; Bi, Rui; Shui, Ruohong; Yu, Baohua; Lu, Hongfen; Cai, Xu; Yang, Wentao

    2014-01-01

    Male breast carcinoma is a relatively rare disease. This study retrospectively investigated the clinicopathological features of 73 cases of male breast carcinoma in Chinese population, and classified the molecular subtype based on surrogate immunohistochemical definitions. The expression of GCDFP15, MGB, AR and FOXP1 were evaluated. Invasive carcinoma of no special type was the most common histological type in the study group (71.2%, 52/73). The luminal A and B subtypes were the major types of male breast carcinoma (60.9%, 34.8% respectively). AR and FOXP1 are expressed in 84.2% (48/57) and 71.9% (41/57) of the studied cases. Carcinoma of the luminal A subtype expressed GCDFP15 (73.5%, 25/34) and MGB (58.8%, 20/34) more frequently than cases of the luminal B subtypes (34.8%, 8/23 and 43.5%, 10/23, respectively; P = 0.004, P = 0.255, respectively). In conclusion, invasive carcinoma of no special type was the most common histological type in male breast carcinoma among Chinese population. Our study revealed that the luminal A and B subtypes were the major types of male breast carcinoma. AR and FOXP1 are highly expressed in male breast cancer. The luminal A subtype tends to express GCDFP15 and MGB more frequently than the luminal B subtype.

  5. Atypical fibroxanthoma: a histological and immunohistochemical review of 171 cases.

    PubMed

    Beer, Trevor W; Drury, Paul; Heenan, Peter J

    2010-08-01

    The clinical and histological features of 171 atypical fibroxanthomas (AFX) from a single institution in Western Australia are outlined. This area experiences high levels of solar radiation, and all assessable biopsies showed solar elastosis. Patients were aged between 41 and 97 years (median age 74), with 76% of tumors occurring in men (male to female ratio approximately 3 to 1). Most tumors were small, with a median diameter of 10 mm and a range of 4-35 mm. Only 5% exceeded 20 mm in diameter. Most AFX were well-circumscribed dermal lesions, with limited invasion of subcutis in a minority. Histological variants identified included keloidal (n = 8), clear cell (n = 3), and granular cell (n = 3), plaque like (n = 4), and myxoid (n = 1). Bland cytological appearances (spindle cell nonpleomorphic AFX) were noted in 5 tumors, with osteoclast-like giant cells in 2. Features suggesting regression were present in 22 cases. Two cases recurred locally, none metastasized. No tumors expressed melanocytic or epithelial markers. Seventy-four percent of cases expressed smooth muscle actin, typically strongly and diffusely. No AFX stained with desmin. Only 1 of 50 cases was CD117 positive. In conclusion, AFX may show a wide range of histological appearances, and a panel of immunohistochemical markers is essential to make the correct diagnosis. Histological mimics, such as poorly differentiated squamous cell carcinoma, must be carefully excluded. Specific diagnosis is important because there seems to be a very low risk of recurrence or metastasis despite the frequently alarming histology. PMID:20526171

  6. Myoepithelioma arising from the buccal gland: histopathological and immunohistochemical studies.

    PubMed

    Sugiura, R; Kuyama, K; Utsunomiya, T; Morikawa, M; Fukumoto, M; Yamamoto, H

    2000-03-01

    A rare case of myoepithelioma of the buccal gland in a 54-year-old Japanese woman is reported. As the swelling exhibited a normal mucosal color and was relatively well defined, showing no ulcers, a benign salivary gland tumor was suspected upon clinical inspection. Microscopically, the parenchyma of the present case mainly consisted of plasmacytoid cells with round nuclei and eosinophilic cytoplasm, and partial spindle cells with eccentric nuclei. The stroma was composed of fibro-hyalinized or myxoid connective tissue that separated from the parenchyma. Immunohistochemically, the cytoplasm of the plasmacytoid and spindle cells was moderately positive for vimentin and GFAP, whereas the buccal gland adjacent to the tumor was negative for these antibodies. S-100 protein reactivity is strong for both types tumor cells. Actin reactivity was negative for both types of tumor cells, notwithstanding the fact that myoepithelial cells of the buccal gland were positively stained. Anti-cytokeratin reactivity was weak for both types of tumor cells in portions of the plexiform and solid areas; nevertheless, the buccal glands were moderately positive. These results suggest that neoplasmic myoepithelial cells exhibit abnormal differentiation and modification. There have been only two published reports of myoepithelioma arising from the buccal gland in the literature to date.

  7. Quantification of immunohistochemical findings of neurofibrillary tangles and senile plaques for a diagnosis of dementia in forensic autopsy cases.

    PubMed

    Takayama, Mio; Kashiwagi, Masayuki; Matsusue, Aya; Waters, Brian; Hara, Kenji; Ikematsu, Natsuki; Kubo, Shin-Ichi

    2016-09-01

    We report the quantification of immunohistochemical findings for a diagnosis of dementia in autopsy cases among older decedents. Autopsy cases were selected with the following requirements: >65yo; no head injuries, thermal injuries, or heat stroke; no intracranial lesions; and within 48h of death. Among cases that met all requirements, 10 had a clinical diagnosis of dementia were included in dementia group. Non-dementia group consisted of 38 cases without any record of dementia. To compare these groups, immunohistochemically, beta-amyloid, tau protein, gephyrin, and IL-33 were examined in five regions. Quantitative analysis was performed by collecting with image data analyzed using analysis software. Image data on tau-immunopositive neurofibrillary tangles (NFT) and beta-amyloid-positive senile plaques (SP) were photographed. Criteria for dementia were made by counting and measuring NFT and SP from image data using software. Differences in SP and NFT were effective for discriminating between the two groups. These criteria may reveal the presence and progression of dementia. Total of tau-positive NFT in Ammon's horn (AH) may be useful for diagnosing dementia. When the total is more than 41 in approximately 6mm(2) of AH, the possibility of dementia is considered. Total of beta-amyloid-positive SP in the parahippocampal gyrus (PHG) may be useful for diagnosing dementia. When the total in approximately 5mm(2) of PHG is more than 47, the possibility of dementia is considered. Immunohistochemical staining may be more useful for obtaining image data for quantification than conventional staining techniques, such as Bielschowsky-Hirano's silver staining. PMID:27591545

  8. Trace element analysis of K, U and Th in high purity materials by neutron activation analysis

    SciTech Connect

    Pillalamarri, Ila

    2005-09-08

    The concept and usage of 'high purity' are explained. Trace element analysis of K, U and Th by neutron activation analysis is described, the radio-isotopes and their corresponding gamma-rays used to identify the elements are listed. The interfering elements are described. The advantages and disadvantages of using neutron activation analysis are discussed. Some examples of trace impurity determinations in high purity materials are provided.

  9. Immunohistochemical localization of D-aspartate oxidase in porcine peripheral tissues.

    PubMed

    Yamamoto, Atsushi; Tanaka, Hiroyuki; Ishida, Tetsuo; Horiike, Kihachiro

    2011-07-01

    D-Aspartate (D-Asp) is an endogenous substance in mammals. Degradation of D-Asp is carried out only by D-aspartate oxidase (DDO). We measured DDO activity in porcine tissues, and produced an anti-porcine DDO antibody to examine the cellular localization of DDO. All the tissues examined showed DDO activities, whereas the substrate D-Asp was not detected in kidney cortex, liver, heart, and gastric mucosa. In the kidney, intensive immunohistochemical staining for DDO was found in the epithelial cells of the proximal tubules. In the liver, the epithelial cells of interlobular bile ducts, liver sinusoid-lining cells with cytoplasmic processes, and the smooth muscle cells of arterioles were strongly stained for DDO. In the heart, cardiomyocytes and the smooth muscle cells of arterioles showed DDO-immunoreactivity. In the gastric mucosa, only the chief cells were DDO-positive. These newly identified DDO-positive cells seem to actively degrade D-Asp to prevent an excess of D-Asp from exerting harmful effects on the respective functions of porcine tissues.

  10. Visual Analysis of Biological Activity Data with Scaffold Hunter.

    PubMed

    Klein, Karsten; Koch, Oliver; Kriege, Nils; Mutzel, Petra; Schäfer, Till

    2013-12-01

    The growing interest in chemogenomics approaches over the last years has led to an increasing amount of data regarding chemical and the corresponding biological activity space. The resulting data, collected in either in-house or public databases, need to be analyzed efficiently to speed-up the increasingly difficult task of drug discovery. Unfortunately, the discovery of new chemical entities or new targets for known drugs ('drug repurposing') is not suitable to a fully automated analysis or a simple drill down process. Visual interactive interfaces that allow to explore chemical space in a systematic manner and facilitate analytical reasoning can help to overcome these problems. Scaffold Hunter is a tool for the visual analysis of chemical compound databases that provides integrated visualization and analysis of biological activity data and fosters the interactive exploration of data imported from a variety of sources. We describe the features and illustrate the use by means of an exemplary analysis workflow.

  11. INDUSTRIAL/MILITARY ACTIVITY-INITIATED ACCIDENT SCREENING ANALYSIS

    SciTech Connect

    D.A. Kalinich

    1999-09-27

    Impacts due to nearby installations and operations were determined in the Preliminary MGDS Hazards Analysis (CRWMS M&O 1996) to be potentially applicable to the proposed repository at Yucca Mountain. This determination was conservatively based on limited knowledge of the potential activities ongoing on or off the Nevada Test Site (NTS). It is intended that the Industrial/Military Activity-Initiated Accident Screening Analysis provided herein will meet the requirements of the ''Standard Review Plan for the Review of Safety Analysis Reports for Nuclear Power Plants'' (NRC 1987) in establishing whether this external event can be screened from further consideration or must be included as a design basis event (DBE) in the development of accident scenarios for the Monitored Geologic Repository (MGR). This analysis only considers issues related to preclosure radiological safety. Issues important to waste isolation as related to impact from nearby installations will be covered in the MGR performance assessment.

  12. Analysis of initial in-plant active neutron multiplicity measurements

    SciTech Connect

    Ensslin, N.; Krick, M.S.; Harker, W.C.

    1993-08-01

    This paper analyzes initial in-plant measurements made by active neutron multiplicity counting, a new technique currently under development for the assay of bulk uranium containing kilograms of {sup 235}U. The measurements were made at Savannah River and Y-12 using active well coincidence counters and prototype multiplicity electronics and software from Los Alamos. For one of the sets of highly enriched uranium samples measured to data, we improved the assay accuracy by an order-of-magnitude by adding the multiplicity analysis to the conventional coincidence analysis. This paper summarizes our results and describes areas where further work is needed.

  13. Physical basis for prompt-neutron activation analysis

    SciTech Connect

    Chrien, R.E.

    1982-01-01

    The technique called prompt ..gamma..-ray neutron activation analysis has been applied to rapid materials analysis. The radiation following the neutron radiation capture is prompt in the sense that the nuclear decay time is on the order of 10/sup -15/ second, and thus the technique is not strictly activation, but should be called radiation neutron capture spectroscopy or neutron capture ..gamma..-ray spectroscopy. This paper reviews the following: sources and detectors, theory of radiative capture, nonstatistical capture, giant dipole resonance, fast neutron capture, and thermal neutron capture ..gamma..-ray spectra. 14 figures.

  14. Overview of MSFC's Applied Fluid Dynamics Analysis Group Activities

    NASA Technical Reports Server (NTRS)

    Garcia, Roberto; Wang, Tee-See; Griffin, Lisa; Turner, James E. (Technical Monitor)

    2001-01-01

    This document is a presentation graphic which reviews the activities of the Applied Fluid Dynamics Analysis Group at Marshall Space Flight Center (i.e., Code TD64). The work of this group focused on supporting the space transportation programs. The work of the group is in Computational Fluid Dynamic tool development. This development is driven by hardware design needs. The major applications for the design and analysis tools are: turbines, pumps, propulsion-to-airframe integration, and combustion devices.

  15. Active Desiccant-Based Preconditioning Market Analysis and Product Development

    SciTech Connect

    Fischer, J.

    2001-01-11

    The Phase 1 report (ORNL/Sub/94-SVO44/1), completed earlier in this program, involved a comprehensive field survey and market analysis comparing various specialized outdoor air handling units. This initial investigation included conventional cooling and reheat, conventional cooling with sensible recovery, total energy recovery systems (passive desiccant technology) and various active desiccant systems. The report concluded that several markets do promise a significant sales opportunity for a Climate Changer-based active desiccant system offering. (Climate Changer is a registered trademark of Trane Company.) This initial market analysis defined the wants and needs of the end customers (design engineers and building owners), which, along with subsequent information included in this report, have been used to guide the determination of the most promising active desiccant system configurations. This Phase 2 report begins with a summary of a more thorough investigation of those specific markets identified as most promising for active desiccant systems. Table 1 estimates the annual sales potential for a cost-effective product line of active desiccant systems, such as that built from Climate Changer modules. The Product Development Strategy section describes the active desiccant system configurations chosen to best fit the needs of the marketplace while minimizing system options. Key design objectives based on market research are listed in this report for these active desiccant systems. Corresponding performance goals for the dehumidification wheel required to meet the overall system design objectives are also defined. The Performance Modeling section describes the strategy used by SEMCO to design the dehumidification wheels integrated into the prototype systems currently being tested as part of the U.S. Department of Energy's Advanced Desiccant Technology Program. Actual performance data from wheel testing was used to revise the system performance and energy analysis

  16. Histochemical and immunohistochemical characterization of chordoma in ferrets

    PubMed Central

    YUI, Takeshi; OHMACHI, Tetsuo; MATSUDA, Kazuya; OKAMOTO, Minoru; TANIYAMA, Hiroyuki

    2015-01-01

    Chordomas of the tip of the tail in 6 ferrets were examined using histopathological, histochemical and immunohistochemical procedures. Histopathologically, round neoplastic cells containing numerous cytoplasmic vacuoles of varying sizes, categorized as “physaliphorous cells”, were observed in the amorphous eosinophilic or pale basophilic myxoid stroma. Physaliphorous cells were arranged in lobules and in a “chordoid” or “cobblestone” manner. The neoplasms were diagnosed as benign chordoma without local invasion and metastasis. Histochemically, the cytoplasm of small neoplastic cells was positive for periodic acid-Schiff stain and alcian blue (AB) pH 2.5 and pH 1.0 stains, but negative for hyaluronidase digestion-AB pH 2.5 stain. All neoplastic cells were strongly stained with colloidal ion, negative for high iron diamine AB pH 2.5 and toluidine blue pH 2.5 stains, and positive for Mayer’s mucicarmine stain. Immunohistochemistry using antibodies directed against low-molecular-weight cytokeratins (CK18, CK19 and CK20), vimentin and mucin core protein (MUC5AC) revealed that neoplastic cells had both epithelial and mesenchymal elements. The expression of low-molecular-weight cytokeratins suggests that neoplastic cells acquired the properties of glandular epithelial cells and produced epithelial mucus. Furthermore, the expression of cytokeratins, vimentin, S100 protein, brachyury and epithelial membrane antigen indicates that the neoplasms were equivalent to the classic type of human chordoma. Therefore, immunohistochemistry using these antibodies can be useful for the characterization of ferret chordoma. PMID:25648567

  17. Immunohistochemical evaluation of myofibroblast density in odontogenic cysts and tumors.

    PubMed

    Kouhsoltani, Maryam; Halimi, Monireh; Jabbari, Golchin

    2016-01-01

    Background. The aim of this study was to investigate myofibroblast (MF) density in a broad spectrum of odontogenic cysts and tumors and the relation between the density of MFs and the clinical behavior of these lesions. Methods. A total of 105 cases of odontogenic lesions, including unicystic ameloblastoma (UAM), solid ameloblastoma (SA), odontogenic keratocyst (OKC), dentigerous cyst (DC), radicular cyst (RC) (15 for each category), and odontogenic myxoma (OM), adenomatoid odontogenic tumor (AOT), calcifying odontogenic cyst (COC) (10 for each category), were immunohistochemically stained with anti-α-smooth muscle actin antibody. The mean percentage of positive cells in 10 high-power fields was considered as MF density for each case. Results. A statistically significant difference was observed in the mean scores between the study groups (P < 0.001). The intensity of MFs was significantly higher in odontogenic tumors compared to odontogenic cysts (P < 0.001). There was no statistically significant difference between odontogenic tumors, except between UAM and OM (P = 0.041). The difference between OKC and odontogenic tumors was not statistically significant (P > 0.05). The number of MFs was significantly higher in OKC and lower in COC compared to other odontogenic cysts (P = 0.007 and P = 0.045, respectively). Conclusion. The results of the present study suggest a role for MFs in the aggressive behavior of odontogenic lesions. MFs may represent an important target of therapy, especially for aggressive odontogenic lesions. Our findings support the classification of OKC in the category of odontogenic tumors. PMID:27092213

  18. Immunohistochemical evaluation of myofibroblast density in odontogenic cysts and tumors

    PubMed Central

    Kouhsoltani, Maryam; Halimi, Monireh; Jabbari, Golchin

    2016-01-01

    Background. The aim of this study was to investigate myofibroblast (MF) density in a broad spectrum of odontogenic cysts and tumors and the relation between the density of MFs and the clinical behavior of these lesions. Methods. A total of 105 cases of odontogenic lesions, including unicystic ameloblastoma (UAM), solid ameloblastoma (SA), odontogenic keratocyst (OKC), dentigerous cyst (DC), radicular cyst (RC) (15 for each category), and odontogenic myxoma (OM), adenomatoid odontogenic tumor (AOT), calcifying odontogenic cyst (COC) (10 for each category), were immunohistochemically stained with anti-α-smooth muscle actin antibody. The mean percentage of positive cells in 10 high-power fields was considered as MF density for each case. Results. A statistically significant difference was observed in the mean scores between the study groups (P < 0.001). The intensity of MFs was significantly higher in odontogenic tumors compared to odontogenic cysts (P < 0.001). There was no statistically significant difference between odontogenic tumors, except between UAM and OM (P = 0.041). The difference between OKC and odontogenic tumors was not statistically significant (P > 0.05). The number of MFs was significantly higher in OKC and lower in COC compared to other odontogenic cysts (P = 0.007 and P = 0.045, respectively). Conclusion. The results of the present study suggest a role for MFs in the aggressive behavior of odontogenic lesions. MFs may represent an important target of therapy, especially for aggressive odontogenic lesions. Our findings support the classification of OKC in the category of odontogenic tumors. PMID:27092213

  19. Immunohistochemical Expression of Leptin in Non Melanoma Skin Cancer

    PubMed Central

    Farag, Azza G.A.; El-Dien, Marwa Mohammed Serag

    2016-01-01

    Introduction Obesity in adults is associated with numerous health disorders including some forms of cancer. Various epidemiological studies have found a link between excess adiposity and malignant melanoma; however, the association with non melanoma skin cancer is questionable. Leptin is a hormone produced mainly by the adipose tissue and its serum level may reflect body mass index. Leptin is reported to promote proliferation and angiogenesis and deregulate apoptosis, therefore facilitates the process of carcinogenesis. Aim The current study tried to assess leptin localization and expression in non melanoma skin cancer to verify its possible role in pathogenesis of this cancer. Materials and Methods This study was carried out on 13 Basal Cell Carcinoma (BCC) cases and 14 Squamous Cell Carcinoma (SCC) cases together with 19 normal skin biopsies as a control group using immunohistochemical method. Results Leptin was expressed in 52.6% of the normal epidermis with pure cytoplasmic and both cytoplasmic and nuclear staining patterns. All cases of SCC (100%) and two cases of BCC (15.4%) showed leptin expression in tumour cells whereas nuclear expression was in favour of SCC. Stromal expression of leptin was seen in both SCC (57.1%) and BCC (38.5%) without significant differences. Percentage of leptin expression by tumour cells in SCC showed positive linear correlation with tumour size (p=0.02) and microvessel density (p=0.000). Stromal expression of leptin in SCC was associated with large tumour size (p=0.04), advanced stage (p=0.01) and tumours arising in sites other than head and neck (p=0.01). Conclusion Leptin could have a more important role in pathogenesis of cutaneous SCC rather than BCC that may reflect the trivial role of obesity in induction of BCC. The expression of leptin by tumour and stromal cells of SCC could co-operate in its progression by promoting angiogenesis with subsequently acquiring large tumour size and then advanced stage. PMID:27656540

  20. Collagenous gastritis: a morphologic and immunohistochemical study of 40 patients.

    PubMed

    Arnason, Thomas; Brown, Ian S; Goldsmith, Jeffrey D; Anderson, William; O'Brien, Blake H; Wilson, Claire; Winter, Harland; Lauwers, Gregory Y

    2015-04-01

    Collagenous gastritis is a rare condition defined histologically by a superficial subepithelial collagen layer. This study further characterizes the morphologic spectrum of collagenous gastritis by evaluating a multi-institutional series of 40 patients (26 female and 14 male). The median age at onset was 16 years (range 3-89 years), including 24 patients (60%) under age 18. Twelve patients (30%) had associated celiac disease, collagenous sprue, or collagenous colitis. Hematoxylin and eosin slides were reviewed in biopsies from all patients and tenascin, gastrin, eotaxin, and IgG4/IgG immunohistochemical stains were applied to a subset. The distribution of subepithelial collagen favored the body/fundus in pediatric patients and the antrum in adults. There were increased surface intraepithelial lymphocytes (>25 lymphocytes/100 epithelial cells) in five patients. Three of these patients had associated celiac and/or collagenous sprue/colitis, while the remaining two had increased duodenal lymphocytosis without specific etiology. An eosinophil-rich pattern (>30 eosinophils/high power field) was seen in 21/40 (52%) patients. Seven patients' biopsies demonstrated atrophy of the gastric corpus mucosa. Tenascin immunohistochemistry highlighted the subepithelial collagen in all 21 specimens evaluated and was a more sensitive method of collagen detection in biopsies from two patients with subtle subepithelial collagen. No increased eotaxin expression was identified in 16 specimens evaluated. One of the twenty-three biopsies tested had increased IgG4-positive cells (100/high power field) with an IgG4/IgG ratio of 55%. In summary, collagenous gastritis presents three distinct histologic patterns including a lymphocytic gastritis-like pattern, an eosinophil-rich pattern, and an atrophic pattern. Eotaxin and IgG4 were not elevated enough to implicate these pathways in the pathogenesis. Tenascin immunohistochemistry can be used as a sensitive method of collagen detection. PMID

  1. Immunohistochemical detection of gastric mucin in normal and disease states.

    PubMed

    Taylor, K L; Mall, A S; Barnard, R A; Ho, S B; Cruse, J P

    1998-01-01

    At least seven human mucin genes have been described, which express glycoproteins MUC1-7 in various tissues. It has been shown that different mucins are expressed in various gastric disease states compared to the normal. In this study we used histochemical and immunohistochemical methods to determine the type and pattern of mucin in 54 patients with a variety of gastric conditions [i.e., normal controls, fetal stomachs, gastritis, low-grade dysplasia, intestinal metaplasia (associated with gastritis, benign ulcers, dysplasia, and cancer), early and advanced intestinal type adenocarcinoma, and diffuse adenocarcinoma]. We report for the first time the use of all seven MUC antibodies in the various conditions. Normal controls were immunoreactive for MUC4, 5, and 6 , and gastritis specimens showed similar results, although the latter showed more MUC1 immunoreactivity. Whereas early fetal stomach showed no MUC immunoreactivity, MUC4, 5, and 6 were present from the early second trimester onwards. There was no significant difference between dysplasia and intestinal metaplasia, both categories showing the presence of MUC2 and 3 predominantly. Early intestinal type adenocarcinomas did not show any mucins in the majority of cases. Advanced intestinal type adenocarcinomas showed immunoreactivity predominantly for MUC1, 5, and 6, as well as MUC2 in some cases. Diffuse adenocarcinomas showed strong positive MUC2 and 6 staining, and in some cases MUC5 and 7. In conclusion, we have shown different patterns of mucin immunoreactivity in various gastric disease states. Specimens with dysplasia, intestinal metaplasia, late intestinal type adenocarcinoma, and diffuse gastric cancer were characterized by increased diversity of mucin types, whereas early intestinal cancer showed loss of mucin immunoreactivity.

  2. Oncocytoma of the salivary glands: a clinicopathologic and immunohistochemical study.

    PubMed

    Zhou, Chuan-Xiang; Gao, Yan

    2009-12-01

    We present a clinicopathologic and immunohistochemical study of 21 Chinese patients with oncocytoma of salivary gland origin, a rare benign tumour composed exclusively of large epithelial cells with eosinophilic granular cytoplasm (oncocytes). The median age was 60.1 years with a male predominance (67%). All the tumours occurred in the parotid except one in the palate. A painless mass was the most common feature, although intermittent pain was complained of in four cases. All the patients were treated by superficial parotidectomy, with no recurrence or metastasis. Histologically, most tumours displayed an encapsulated nodular growth pattern, but one case presented with an aggressive growing tendency. Typical oncocytes were observed in all cases, with one clear cell variant found. The oncocytes were arranged in solid sheets, trabecular or duct-like structures. Rarely, small foci of hemorrhage or lymphoid stroma were observed, but germinal centres were always absent. Phosphotungstic acid hematoxylin staining illustrated dark-blue cytoplasmic granules, demonstrated as mitochondria by electron microscopy. All the tumours showed immunoreactivity for CK5/6, CK8/18, CK10/13, CK19 and EMA, but were negative for SMA or S-100. MIB-1 antibody, used to identify the dividing cells by staining of the nucleus, was found to stain the cytoplasm of the oncocytes. In summary, clinical diagnosis for an oncocytoma is challenging for its similar features to other benign tumours. Histopathological diagnosis is reliable with histochemical and electron microscopic conformation of the oncocytes, but differential diagnosis is still challenging. MIB-1 immunostaining might be considered as a diagnostic aid.

  3. GATA3 immunohistochemical expression in salivary gland neoplasms.

    PubMed

    Schwartz, Lauren E; Begum, Shahnaz; Westra, William H; Bishop, Justin A

    2013-12-01

    GATA3 is a zinc finger transcription factor that regulates the normal development of many tissues and cell types. Recent studies have shown that immunohistochemical nuclear staining for GATA3 among tumors is highly restricted to carcinomas of breast and urothelial origin; however salivary gland tumors have not been tested. Given that breast and salivary gland tissues are very similar with respect to embryologic development and structure, we performed GATA3 staining on a spectrum of salivary gland neoplasms. GATA3 immunohistochemistry was performed on a diverse collection of 180 benign and malignant salivary gland neoplasms including 10 acinic cell carcinomas, 2 adenocarcinomas not otherwise specified, 41 adenoid cystic carcinomas, 2 epithelial-myoepithelial carcinomas, 1 low grade cribriform cystadenocarcinoma, 15 mammary analogue secretory carcinomas, 7 metastatic squamous cell carcinomas, 27 mucoepidermoid carcinomas, 2 oncocytic carcinomas, 5 oncocytomas, 34 pleomorphic adenomas, 4 polymorphous low grade adenocarcinomas, 25 salivary duct carcinomas, and 5 Warthin tumors. Staining for GATA3 was observed in 92/180 (51 %) of salivary gland tumors. GATA3 staining was observed in most of the tumor types, but diffuse immunolabeling was consistently seen in salivary duct carcinoma (25 of 25) and mammary analogue secretory carcinoma (15 of 15)-the two tumor types that most closely resemble breast neoplasia. Background benign salivary gland tissue was also usually weakly positive in both acini and ducts. GATA3 immunostaining is not restricted to tumors of breast and urothelial origin. Rather, it is expressed across many different types of salivary gland neoplasms. As a result, salivary gland origin should be considered in the differential diagnosis of a GATA3-positive carcinoma, particularly in the head and neck. Although GATA3 immunohistochemistry is not helpful in resolving the differential diagnosis between a primary salivary gland neoplasm and metastatic breast

  4. Sialoblastoma: a clinicopathologic and immunohistochemical study of 7 cases.

    PubMed

    Williams, Stephen B; Ellis, Gary L; Warnock, Gary R

    2006-12-01

    Sialoblastoma is a rare congenital or perinatal salivary tumor that varies in histologic features and biologic potential. Seven cases from the files of the Armed Forces Institute of Pathology are presented. These tumors occurred in 4 males and 3 females with ages ranging from prenatal to 6 months at the time of discovery. Five lesions originated from the parotid gland; 2 lesions were from the submandibular gland. All lesions presented as nodular to multinodular swellings and ranged in size from 2.0 to 7.0 cm. The principal sign or symptom was rapid growth. Two histologic patterns with differing behavior predominated: (1) a favorable pattern had semiencapsulation of cytologically benign basaloid tumor cells with intervening stroma; and (2) an unfavorable histology of anaplastic basaloid tumor cells, minimal stroma, and broad pushing to infiltrative periphery. Four and three tumors had favorable and unfavorable growth patterns, respectively. One unfavorable lesion had vascular invasion, and another demonstrated perineural invasion. All 3 tumors with unfavorable histology recurred. Tumor cells in 3 cases were immunohistochemically reactive for keratin, S-100, smooth muscle actin, and calponin to varying degrees. All 3 tumors were reactive for p63. alpha-Fetoprotein was expressed in 2 unfavorable tumors. Ki67 was expressed at 3% in a favorable tumor and 40% and 80% in the 2 unfavorable lesions. Treatment involved surgical excision. One patient received adjuvant chemotherapy. Two sialoblastomas resulted in recurrences within a year and another developed a recurrence after 4 years. One sialoblastoma developed lung metastasis within 1 month of the original biopsy. Although a clinical correlation is suggested by a favorable/unfavorable histologic grading system the biologic behavior is nonetheless considered unpredictable.

  5. Preliminary Work Domain Analysis for Human Extravehicular Activity

    NASA Technical Reports Server (NTRS)

    McGuire, Kerry; Miller, Matthew; Feigh, Karen

    2015-01-01

    A work domain analysis (WDA) of human extravehicular activity (EVA) is presented in this study. A formative methodology such as Cognitive Work Analysis (CWA) offers a new perspective to the knowledge gained from the past 50 years of living and working in space for the development of future EVA support systems. EVA is a vital component of human spaceflight and provides a case study example of applying a work domain analysis (WDA) to a complex sociotechnical system. The WDA presented here illustrates how the physical characteristics of the environment, hardware, and life support systems of the domain guide the potential avenues and functional needs of future EVA decision support system development.

  6. Validation of in vivo pharmacodynamic activity of a novel PDGF receptor tyrosine kinase inhibitor using immunohistochemistry and quantitative image analysis.

    PubMed

    D'Andrea, Michael R; Mei, Jay M; Tuman, Robert W; Galemmo, Robert A; Johnson, Dana L

    2005-08-01

    With the advent of agents directed against specific molecular targets in drug discovery, it has become imperative to show a compound's cellular impact on the intended biomolecule in vivo. The objective of the present study was to determine if we could develop an assay to validate the in vivo effects of a compound. Hence, we investigated the in vivo pharmacodynamic activity of JNJ-10198409, a relatively selective inhibitor of platelet-derived growth factor receptor tyrosine kinase (PDGF-RTK), in tumor tissues after administering the compound orally in a nude mouse xenograft model of human LoVo colon cancer. We developed a novel assay to quantify the in vivo anti-PDGF-RTK activity of the inhibitor in tumor tissue by determining the phosphorylation status of phospholipase Cgamma1 (PLCgamma1), a key downstream cellular molecule in the PDGF-RTK signaling cascade. We used two antibodies, one specific for the total (phosphorylated and unphosphorylated forms) PLCgamma1 (pan-PLCgamma1) and the other, specific for phosphorylated form of PLCgamma1 (ph-PLCgamma1) to immunohistochemically detect their expression in tumor tissues. Computer-assisted image analysis was then used to directly compare the ratio of ph-PLCgamma1 to pan-PLCgamma1 immunolabeling intensities in serial sections (5 mum) of tumors obtained from vehicle- and JNJ-10198409-treated tumor-bearing mice. Our data showed statistically significant, dose-dependent differences in the ph-PLC/pan-PLC ratio among the four treatment groups (vehicle, 25, 50, and 100 mg/kg b.i.d.). These results confirmed this compound's ability to suppress PDGF-RTK downstream signaling in tumor tissues in vivo. In addition to this specific application of this in vivo validation approach to those targets that use PLCgamma as a downstream signaling partner, these methods may also benefit other drug discovery targets. PMID:16093435

  7. Optimizing human activity patterns using global sensitivity analysis

    PubMed Central

    Hickmann, Kyle S.; Mniszewski, Susan M.; Del Valle, Sara Y.; Hyman, James M.

    2014-01-01

    Implementing realistic activity patterns for a population is crucial for modeling, for example, disease spread, supply and demand, and disaster response. Using the dynamic activity simulation engine, DASim, we generate schedules for a population that capture regular (e.g., working, eating, and sleeping) and irregular activities (e.g., shopping or going to the doctor). We use the sample entropy (SampEn) statistic to quantify a schedule’s regularity for a population. We show how to tune an activity’s regularity by adjusting SampEn, thereby making it possible to realistically design activities when creating a schedule. The tuning process sets up a computationally intractable high-dimensional optimization problem. To reduce the computational demand, we use Bayesian Gaussian process regression to compute global sensitivity indices and identify the parameters that have the greatest effect on the variance of SampEn. We use the harmony search (HS) global optimization algorithm to locate global optima. Our results show that HS combined with global sensitivity analysis can efficiently tune the SampEn statistic with few search iterations. We demonstrate how global sensitivity analysis can guide statistical emulation and global optimization algorithms to efficiently tune activities and generate realistic activity patterns. Though our tuning methods are applied to dynamic activity schedule generation, they are general and represent a significant step in the direction of automated tuning and optimization of high-dimensional computer simulations. PMID:25580080

  8. Optimizing human activity patterns using global sensitivity analysis

    SciTech Connect

    Fairchild, Geoffrey; Hickmann, Kyle S.; Mniszewski, Susan M.; Del Valle, Sara Y.; Hyman, James M.

    2013-12-10

    Implementing realistic activity patterns for a population is crucial for modeling, for example, disease spread, supply and demand, and disaster response. Using the dynamic activity simulation engine, DASim, we generate schedules for a population that capture regular (e.g., working, eating, and sleeping) and irregular activities (e.g., shopping or going to the doctor). We use the sample entropy (SampEn) statistic to quantify a schedule’s regularity for a population. We show how to tune an activity’s regularity by adjusting SampEn, thereby making it possible to realistically design activities when creating a schedule. The tuning process sets up a computationally intractable high-dimensional optimization problem. To reduce the computational demand, we use Bayesian Gaussian process regression to compute global sensitivity indices and identify the parameters that have the greatest effect on the variance of SampEn. Here we use the harmony search (HS) global optimization algorithm to locate global optima. Our results show that HS combined with global sensitivity analysis can efficiently tune the SampEn statistic with few search iterations. We demonstrate how global sensitivity analysis can guide statistical emulation and global optimization algorithms to efficiently tune activities and generate realistic activity patterns. Finally, though our tuning methods are applied to dynamic activity schedule generation, they are general and represent a significant step in the direction of automated tuning and optimization of high-dimensional computer simulations.

  9. Optimizing human activity patterns using global sensitivity analysis

    DOE PAGES

    Fairchild, Geoffrey; Hickmann, Kyle S.; Mniszewski, Susan M.; Del Valle, Sara Y.; Hyman, James M.

    2013-12-10

    Implementing realistic activity patterns for a population is crucial for modeling, for example, disease spread, supply and demand, and disaster response. Using the dynamic activity simulation engine, DASim, we generate schedules for a population that capture regular (e.g., working, eating, and sleeping) and irregular activities (e.g., shopping or going to the doctor). We use the sample entropy (SampEn) statistic to quantify a schedule’s regularity for a population. We show how to tune an activity’s regularity by adjusting SampEn, thereby making it possible to realistically design activities when creating a schedule. The tuning process sets up a computationally intractable high-dimensional optimizationmore » problem. To reduce the computational demand, we use Bayesian Gaussian process regression to compute global sensitivity indices and identify the parameters that have the greatest effect on the variance of SampEn. Here we use the harmony search (HS) global optimization algorithm to locate global optima. Our results show that HS combined with global sensitivity analysis can efficiently tune the SampEn statistic with few search iterations. We demonstrate how global sensitivity analysis can guide statistical emulation and global optimization algorithms to efficiently tune activities and generate realistic activity patterns. Finally, though our tuning methods are applied to dynamic activity schedule generation, they are general and represent a significant step in the direction of automated tuning and optimization of high-dimensional computer simulations.« less

  10. Aberrant PLAG1 expression in pleomorphic adenomas of the salivary gland: a molecular genetic and immunohistochemical study.

    PubMed

    Matsuyama, Atsuji; Hisaoka, Masanori; Nagao, Yuichi; Hashimoto, Hiroshi

    2011-05-01

    The morphologic distinction of pleomorphic adenoma from other benign or low-grade salivary gland tumors is sometimes difficult and problematic because of their potentially overlapping histological patterns. A subset of pleomorphic adenoma harbors specific gene alterations involving PLAG1 or HMGA2, and the detection of these fusion genes and their products using formalin-fixed, paraffin-embedded (FFPE) tumor specimens may be a useful diagnostic adjunct. In the present study, gene fusions involving PLAG1 or HMGA2 were examined by reverse transcription-polymerase chain reaction (RT-PCR) analysis, with FFPE tumor tissues and immunohistochemical expression of PLAG1 in 45 pleomorphic adenomas, using a commercially available antibody. RT-PCR analyses identified the CTNNB1-PLAG1, LIFR-PLAG1, CHCHD7-PLAG1, and HMGA2-WIF1 fusion transcripts in eight, two, one, and one case, respectively. The TCEA1-PLAG1, HMGA2-FHIT, and HMGA2-NFIB fusion transcripts were not detected. Immunohistochemically, tumor cells in all 45 pleomorphic adenomas were positive for PLAG1, irrespective of PLAG1 rearrangements, even in the case with the HMGA2-WIF1 fusion transcript. Tumor cells displaying myoepithelial or cartilaginous differentiation were almost constantly positive for PLAG1, whereas a limited expression was observed in glandular or keratinizing cells. Among the 46 tumors other than pleomorphic adenoma, 4 carcinomatous components of carcinomas ex pleomorphic adenoma were positive for PLAG1, the other 39 were negative for PLAG1, and the remaining 3 were only faintly and/or focally stained, indicating that the immunohistochemical detection of PLAG1 is diagnostically useful. The present results also suggest that overexpression of PLAG1 is essential for the tumorigenesis of pleomorphic adenomas, although the mechanisms mediating PLAG1 overexpression seem to be variable.

  11. Characterization and immunohistochemical localization of the glycoconjugates of the rabbit bladder mucosa.

    PubMed

    Buckley, M S; Washington, S; Laurent, C; Erickson, D R; Bhavananadan, V P

    1996-06-01

    An impairment of the mucosal glycoconjugates could be an important factor in the development of bladder disorders such as interstitial cystitis. However, very little definitive biochemical information is available on the glycoco-jugate components of the mammalian bladder mucosa. In this-study, the mucosa from metabolically radiolabeled rabbit bladder was separated, delipidated, and digested with protease, and the released glycosaminoglycans and glycopeptides were fractionated. About 80 and 36% of the nondialyzable tritium and 35S activities, respectively, was associated with the sialoglycopeptide fractions. The balance of the total tritium activity in the protease digest was in glycosaminoglycans identified as hyaluronan, chondroitin sulfates, and heparan sulfate. Immunohistochemical examination using anti-heparan sulfate antibodies, including one against mouse syndecan-1, indicated the presence of heparan sulfate proteoglycan in the epithelium. In contrast, there was no significant staining of the bladder epithelium with anti-chondroitin-4- and 6-sulfate antibodies or hyaluronan-binding protein. The lamina propria and muscle layers showed strong staining with anti-chondroitin-4-sulfate antibody and hyaluronan-binding protein and weak staining with anti-chondroitin-6-sulfate antibody. The insignificant levels of glycosaminoglycans in the glycocalyx of bladder mucosa epithelium suggest that glycosaminoglycans may be less important than other glycoconjugates in maintaining normal epithelial function and in bladder disorders such as interstitial cystitis.

  12. Statistical Analysis of Acoustic Wave Parameters Near Solar Active Regions

    NASA Astrophysics Data System (ADS)

    Rabello-Soares, M. Cristina; Bogart, Richard S.; Scherrer, Philip H.

    2016-08-01

    In order to quantify the influence of magnetic fields on acoustic mode parameters and flows in and around active regions, we analyze the differences in the parameters in magnetically quiet regions nearby an active region (which we call “nearby regions”), compared with those of quiet regions at the same disk locations for which there are no neighboring active regions. We also compare the mode parameters in active regions with those in comparably located quiet regions. Our analysis is based on ring-diagram analysis of all active regions observed by the Helioseismic and Magnetic Imager (HMI) during almost five years. We find that the frequency at which the mode amplitude changes from attenuation to amplification in the quiet nearby regions is around 4.2 mHz, in contrast to the active regions, for which it is about 5.1 mHz. This amplitude enhacement (the “acoustic halo effect”) is as large as that observed in the active regions, and has a very weak dependence on the wave propagation direction. The mode energy difference in nearby regions also changes from a deficit to an excess at around 4.2 mHz, but averages to zero over all modes. The frequency difference in nearby regions increases with increasing frequency until a point at which the frequency shifts turn over sharply, as in active regions. However, this turnover occurs around 4.9 mHz, which is significantly below the acoustic cutoff frequency. Inverting the horizontal flow parameters in the direction of the neigboring active regions, we find flows that are consistent with a model of the thermal energy flow being blocked directly below the active region.

  13. Immunohistochemical characterisation of molecular subtypes in endometrial cancer

    PubMed Central

    Łapińska-Szumczyk, Sylwia M; Supernat, Anna M; Majewska, Hanna I; Gulczyński, Jacek; Biernat, Wojciech; Wydra, Dariusz; Żaczek, Anna J

    2015-01-01

    Four molecular subtypes have lately been established in endometrial cancer basing on estrogen receptor (ER), progesterone receptor (PR) and HER2 status: ER+/PR+/HER2+, ER+/PR+/HER2-, ER-/PR-/HER2+ and ER-/PR-/HER2-. The subtypes have shown diversity in terms of prognosis, clinicopathological and molecular characteristics, with ER+/PR+/HER2- and ER-/PR-/HER2+ group exhibiting exceptionally benign and aggressive behavior, respectively. We have further characterized the subtypes in the context of pathways known to drive endometrial carcinogenesis: phosphatidylinositol 3-kinase (PI3K)-AKT pathway (ERBB/PI3K pathway), TP53 system, and the mismatch repair (MMR) mechanism. Analysis of tumor heterogeneity was also included. ER+/PR+/HER2+ was characterized by active ERBB/PI3K pathway occurring in 58% of cases. Subtype ER-/PR-/HER2+ was characterized by the most frequent TP53 mutations (83% of cases). Triple negative phenotype utterly lacked active ERBB/PI3K pathway. Analyzed major pathways rarely correlated with clinicopathologial data but mutated TP53 and retained MMR did correlate with shorter overall survival (both P<0.01). The presence of tumor heterogeneity was most frequent in ER-/PR-/HER2+ subtype (53% of all cases). The presented results further emphasize that the molecular subtype distinction, along with MMR and TP53 status, could be a useful diagnostic tool in guiding individualized therapy. PMID:26885170

  14. Lipomas of the oral cavity: clinicopathological and immunohistochemical study of 24 cases and review of the literature.

    PubMed

    Naruse, Tomofumi; Yanamoto, Souichi; Yamada, Shin-Ichi; Rokutanda, Satoshi; Kawakita, Akiko; Takahashi, Hidenori; Matsushita, Yuki; Hayashida, Saki; Imayama, Naomi; Morishita, Kota; Yamashita, Kentaro; Kawasaki, Goro; Umeda, Masahiro

    2015-03-01

    Although lipomas are common soft tissue tumors, few cases of lipoma or its variants have been reported in the oral cavity. We here described the clinical, histological, and immunohistochemical features of 24 cases of oral lipoma obtained from medical records at Nagasaki University Hospital between 1977 and 2010, and also retrospectively reviewed 603 cases of oral lipoma reported in the English literatures. The patients examined comprised 11 men and 13 women with a mean age of 59 years, ranging from 31 to 90 years. The main sites involved were the buccal mucosa (n = 9), followed by the tongue (n = 4), lip and retromolar area (n = 3), floor of the mouth (n = 2), and gingiva (n = 1). The mean tumor size was 2.0 cm, ranging from 0.2 to 5 cm. Histological analysis revealed 20 cases of lipoma, 2 cases of fibrolipoma, and one case each of intramuscular lipoma and spindle cell lipoma. Twenty-three cases were treated surgically while one case underwent biopsy and follow-up. Recurrence was not observed in any case. We reviewed the English literatures, and similar results were obtained. In immunohistochemical analysis, PCNA and ki-67 expression indices were higher in intramuscular lipoma cases than in its variants. Especially, it showed that a long time follow-up may be necessary in ki-67 positive cases.

  15. Quantitative Assessment of Tumor Associated Macrophages in Head and Neck Squamous Cell Carcinoma Using CD68 Marker: An Immunohistochemical Study

    PubMed Central

    Bagul, Neeta; Roy, Souparna; Ganjre, Anjali; Meher, Aishwarya; Singh, Pratibha

    2016-01-01

    Introduction Oral Squamous Cell Carcinoma (OSCC) is one of the most prevalent cancers in India. Clear evidence regarding inflammation being an etiological factor of cancer was found only in the last few decades. A major inflammatory component in the tumor tissue is Tumor-Associated Macrophages (TAMs). The CD68 antibody is a marker for staining TAMs. Aim The aim of this study is to quantify the macrophage count in healthy oral mucosa and OSCC and comparing TAMs in different histopathological grades of OSCC immunohistochemically. Materials and Methods Thirty archival specimens of OSCC patients and 10 healthy biopsy samples were collected. Immunohistochemical staining was done using a CD68 marker. Statistical analysis was done using Kruskal-Wallis ANOVA and Mann-Whitney U test. Results Comparing CD68 expression in various study groups showed a significant difference (p=0.000). The pair-wise analysis showed different grades of OSCC, which differed significantly for CD68 expression from the normal oral mucosa. Conclusion The most significant cells present in tumor stroma are TAMs, which remain in close proximity to neoplastic cells and interact with them via several chemical mediators, which may serve to increase the invasiveness of the malignant epithelium. Dense infiltration of TAMs adjacent to tumor cells and islands vividly implies their role in tumor progression. PMID:27190959

  16. The Analysis and Reconciliation of Students' Rebuttals in Argumentation Activities

    ERIC Educational Resources Information Center

    Lin, Yu-Ren; Hung, Jeng-Fung

    2016-01-01

    The present study investigated the guidance provided by science teachers to resolve conflicts during socioscientific issue-based argumentation activities. A graphical representation (GR) was developed as a tool to code and analyze the dialogue interaction process. Through the GR and qualitative analysis, we identified three types of dialogue…

  17. Making Sense of Total VET Activity: An Initial Market Analysis

    ERIC Educational Resources Information Center

    National Centre for Vocational Education Research (NCVER), 2016

    2016-01-01

    Following the successful first national publication of total vocational education and training (VET) activity and presentation of various informative data products, NCVER has continued to undertake further analysis of the submitted data. This paper is the first in a suite of the National Centre for Vocational Education Research (NCVER) authored…

  18. Pupil Inquiry Behavior Analysis and Change Activity. Interim Project Report.

    ERIC Educational Resources Information Center

    Manion, Raymond C.

    This interim report discusses progress toward three major goals of the Pupil Inquiry Behavior Analysis and Change Activity: increased pupil inquiry, changed teacher behavior to facilitate pupil inquiry, and the development of a 32-week course of instruction to provide for these behavioral changes. Data currently available deals with the emotional…

  19. Concurrent Validity For an Activity Vector Analysis Index of Anxiety.

    ERIC Educational Resources Information Center

    Plante, Thomas G.

    1982-01-01

    Utilizing 182 subjects, this study determined the concurrent validity between the Activity Vector Analysis (AVA) index of anxiety and scores on the IPAT Anxiety Scale. The IPAT and the AVA index of anxiety seem essentially to measure the same construct of basic anxiety. (Author/PN)

  20. Automated Ki-67 Quantification of Immunohistochemical Staining Image of Human Nasopharyngeal Carcinoma Xenografts

    PubMed Central

    Shi, Peng; Zhong, Jing; Hong, Jinsheng; Huang, Rongfang; Wang, Kaijun; Chen, Yunbin

    2016-01-01

    Nasopharyngeal carcinoma is one of the malignant neoplasm with high incidence in China and south-east Asia. Ki-67 protein is strictly associated with cell proliferation and malignant degree. Cells with higher Ki-67 expression are always sensitive to chemotherapy and radiotherapy, the assessment of which is beneficial to NPC treatment. It is still challenging to automatically analyze immunohistochemical Ki-67 staining nasopharyngeal carcinoma images due to the uneven color distributions in different cell types. In order to solve the problem, an automated image processing pipeline based on clustering of local correlation features is proposed in this paper. Unlike traditional morphology-based methods, our algorithm segments cells by classifying image pixels on the basis of local pixel correlations from particularly selected color spaces, then characterizes cells with a set of grading criteria for the reference of pathological analysis. Experimental results showed high accuracy and robustness in nucleus segmentation despite image data variance. Quantitative indicators obtained in this essay provide a reliable evidence for the analysis of Ki-67 staining nasopharyngeal carcinoma microscopic images, which would be helpful in relevant histopathological researches. PMID:27562647

  1. Quantitative comparison of angiogenesis and lymphangiogenesis in cutaneous lichen planus and psoriasis: immunohistochemical assessment.

    PubMed

    Výbohová, Desanka; Mellová, Yvetta; Adamicová, Katarína; Adamkov, Marián; Hešková, Gabriela

    2015-01-01

    Recent experimental studies revealed that angiogenesis and lymphangiogenesis are closely related to chronic inflammation. The present study aims to evaluate quantitative changes of blood and lymphatic microcirculatory beds in cutaneous lichen planus (CLP) and psoriatic lesions using immunohistochemical analysis with antibodies to CD34, D2-40 and VEGF. Morphometric software was used to determine the area of blood and lymphatic vessels (BVA and LVA) and also the VEGF positive area. Statistical analysis of these parameters confirmed a significant enlargement of both the blood and lymphatic microcirculatory beds in psoriatic and CLP lesions. BVA in CLP lesions was increased by 56% however this augmentation was not as great as in psoriatic lesions where BVA was increased by 123%. Interestingly, LVA in psoriatic and CLP lesions was increased equally by 85%. The strongest VEGF expression was detected in psoriatic lesions, with lower, but still significant, overexpression in CLP lesions. VEGF-C was significantly increased in both psoriatic and CLP lesions in comparable level. Noticeably higher VEGF and VEGF-C expression was observed in the epidermis than in the dermis. Finally, our results indicate that the level of angiogenesis is considerably greater in psoriatic lesions than in CLP lesions, but the level of lymphangiogenesis is equal in both psoriatic and CLP lesions. PMID:25466990

  2. Quantitative immunohistochemical assessment of blood and lymphatic microcirculation in cutaneous lichen planus lesions.

    PubMed

    Výbohová, Desanka; Mellová, Yvetta; Adamicová, Katarína; Adamkov, Marián; Hešková, Gabriela

    2015-06-01

    Latest advances have brought to light the hypothesis that angiogenesis and lymphangiogenesis are tightly connected to some chronic inflammatory diseases. The present study focuses on immunohistochemical assessment of the quantitative changes in the blood and lymphatic microcirculatory bed in common chronic dermatosis - cutaneous lichen planus. Double immunohistochemistry with CD34 and podoplanin antibodies was used to detect blood and lymphatic endothelium, while anti-human VEGF was used for the observation of a key angiogenesis and lymphangiogenesis inducer. Morphometric analysis was performed with QuickPhoto Micro image analysis software. Results confirmed statistically significant enlargement of both the blood and lymphatic microcirculatory beds. Compared to healthy skin, cutaneous lichen planus lesions revealed 1.6 times enlarged blood microcirculatory bed and 1.8 times enlarged lymphatic microcirculatory bed. Vascular endothelial growth factor (VEGF) expression in lesional skin was significantly higher in the epidermis (19.1 times increase) than in the dermis (10.3 times increase). These findings indicate a tight association of angiogenesis and lymphangiogenesis with the pathogenesis of cutaneous lichen planus. PMID:25504638

  3. Estrogen and progesterone receptors in breast cancer. Immunohistochemical assay on scraping material.

    PubMed

    Frigo, B; Pilotti, S; Coradini, D; La Malfa, G; Rilke, F

    1992-04-01

    In order to demonstrate the reliability of immunocytochemical results on cytologic specimens for receptor analysis, the expression of estrogen and progesterone receptors was investigated using immunohistochemistry on frozen sections and on scraping material from the same samples of 50 breast carcinomas. The level of agreement between the two procedures was evaluated by the kappa statistic, as was that between each immunohistochemical procedure and the dextran-coated-charcoal assay since the latter is still the assay employed most frequently for steroid receptor determination and is used for official reports. Statistical results revealed very good agreement regarding the estrogen receptor analysis, with kappa values of .910 and .952 for the comparison of the dextran-coated-charcoal assay with immunocytochemistry on frozen sections and on scrapes, respectively, and .950 for the comparison between the two immunocytochemical procedures. As to progesterone receptors, the kappa values were .795 and .712 for the comparison between the biochemical and immunocytochemical results and .915 for agreement evaluation between the two immunocytochemical procedures. The study showed that the scraping procedure is a valuable tool for the immunocytochemical assessment of steroid receptors in small mammary tumors; it yields representative cellular samples, thus permitting the investigation of heterogeneously distributed substances in tissues.

  4. Acute injury affects lubricin expression in knee menisci: an immunohistochemical study.

    PubMed

    Musumeci, Giuseppe; Loreto, Carla; Carnazza, Maria Luisa; Cardile, Venera; Leonardi, Rosalia

    2013-03-01

    The aim of this study was to investigate for the first time lubricin expression in intact menisci and in menisci from patients with recent knee joint injury using histology, immunohistochemistry, Western blotting and gene expression analysis, to provide insights into pathological processes affecting meniscal tissue. Lubricin expression was studied in vivo in 20 patients (14 males and 6 females) with recent joint injury subjected to arthroscopic partial meniscectomy and in vitro in fibroblast-like cells from meniscus tissue to establish whether it is down-regulated following acute traumatic knee injury. The control group consisted of cadaver donors with normal menisci. Histology demonstrated a normal tissue without structural changes in control samples and structural alterations and clefts in injured menisci. Very strong lubricin immunohistochemical staining was observed in intact menisci; in contrast weak staining was seen in injured menisci. Western blot and mRNA expression analysis also demonstrated strong lubricin expression in control cells and a negligible amount of lubricin in injured fibroblast-like cells. Our data provide information concerning the immediate in vivo response to injury of human knee menisci by documenting early changes in the boundary-lubricating ability of synovial fluid and articular cartilage integrity. These findings may provide the biological basis for developing novel medical therapies to be applied before surgical treatment to preserve tissue function and prevent cartilage damage.

  5. Severe necrotizing encephalitis in a Yorkshire terrier: topographic and immunohistochemical study.

    PubMed

    Lezmi, S; Toussaint, Y; Prata, D; Lejeune, T; Ferreira-Neves, P; Rakotovao, F; Fontaine, J J; Marchal, T; Cordonnier, N

    2007-05-01

    Necrotizing encephalitis of the Yorkshire terrier is a chronic non-suppurative encephalitis that was reported in approximately 15 cases worldwide. We report the case of a 10-year-old female Yorkshire terrier with gross evidence of severe cortical degeneration and necrosis. Microscopically, affected areas were mainly located in the cortical white matter and in the mesencephalon without implication of the cerebellum. Cavitation necrosis, demyelination, gemistocytic astrocytosis, marked perivascular lymphocytic cuffing with a diffuse lymphocytic/histiocytic/gitter cell infiltration characterized the lesions. Immunohistochemical analysis identified the major infiltration of T lymphocytes and macrophages with implication of some cytotoxic lymphocytes and IgG-producing plasma cells; depositions of IgG in the affected white matter were also observed. Specific stains did not reveal fungal, protozoal or bacterial organisms and reverse transcriptase-polymerase chain reaction analysis for distemper virus was also negative. The lympho-histiocytic inflammation suggests a T-cell-mediated and a delayed-type immune reaction as a possible pathogenic mechanism for this brain disorder.

  6. Automated Ki-67 Quantification of Immunohistochemical Staining Image of Human Nasopharyngeal Carcinoma Xenografts.

    PubMed

    Shi, Peng; Zhong, Jing; Hong, Jinsheng; Huang, Rongfang; Wang, Kaijun; Chen, Yunbin

    2016-01-01

    Nasopharyngeal carcinoma is one of the malignant neoplasm with high incidence in China and south-east Asia. Ki-67 protein is strictly associated with cell proliferation and malignant degree. Cells with higher Ki-67 expression are always sensitive to chemotherapy and radiotherapy, the assessment of which is beneficial to NPC treatment. It is still challenging to automatically analyze immunohistochemical Ki-67 staining nasopharyngeal carcinoma images due to the uneven color distributions in different cell types. In order to solve the problem, an automated image processing pipeline based on clustering of local correlation features is proposed in this paper. Unlike traditional morphology-based methods, our algorithm segments cells by classifying image pixels on the basis of local pixel correlations from particularly selected color spaces, then characterizes cells with a set of grading criteria for the reference of pathological analysis. Experimental results showed high accuracy and robustness in nucleus segmentation despite image data variance. Quantitative indicators obtained in this essay provide a reliable evidence for the analysis of Ki-67 staining nasopharyngeal carcinoma microscopic images, which would be helpful in relevant histopathological researches. PMID:27562647

  7. Histopathological and immunohistochemical findings of primary and metastatic medullary thyroid carcinoma in a young dog

    PubMed Central

    Vieson, Miranda D.; Ramos-Vara, José A.; Moon-Larson, Martha; Saunders, Geoffrey

    2014-01-01

    This report describes the gross, histological, and immunohistochemical features of medullary thyroid carcinoma (MTC) with pulmonary metastases in a young dog. Sheets of pleomorphic cells supported by fibrous stroma characterized the primary mass, while metastatic nodules had a neuroendocrine pattern. Despite differing histologic features, all masses showed marked immunoreactivity against calcitonin and multiple neuroendocrine markers consistent with MTC. Although MTC is a well-recognized entity, it may be difficult to distinguish this mass from other thyroid neoplasms, necessitating immunohistochemical characterization. PMID:24690600

  8. neutron activation analysis using thermochromatography. II. thermochromatographic separation of elements in the analysis of geological samples

    SciTech Connect

    Sattarov, G.; Davydov, A.V.; Khamatov, S.; Kist, A.A.

    1986-07-01

    The use of gas thermochromatography (GTC) in the radioactivation analysis of difficulty soluble samples with a strongly activating substrate is discussed. The effect of sample coarseness and ore type on the rate of extraction of gold and accompanying elements was studied. The limits of detection of 22 elements were compared using neutron activation analysis with GTC and INAA. The analytical parameters of the procedure were estimated.

  9. Quantitative analysis of cardiovascular modulation in respiratory neural activity.

    PubMed

    Dick, Thomas E; Morris, Kendall F

    2004-05-01

    We propose the 'delta(2)-statistic' for assessing the magnitude and statistical significance of arterial pulse-modulated activity of single neurones and present the results of applying this tool to medullary respiratory-modulated units. This analytical tool is a modification of the eta(2)-statistic and, consequently, based on the analysis of variance. The eta(2)-statistic reflects the consistency of respiratory-modulated activity on a cycle-by-cycle basis. However, directly applying this test to activity during the cardiac cycle proved ineffective because subjects-by-treatments matrices did not contain enough 'information'. We increased information by dividing the cardiac cycle into fewer bins, excluding cycles without activity and summing activity over multiple cycles. The analysed neuronal activity was an existing data set examining the neural control of respiration and cough. Neurones were recorded in the nuclei of the solitary tracts, and in the rostral and caudal ventral respiratory groups of decerebrate, neuromuscularly blocked, ventilated cats (n= 19). Two hundred of 246 spike trains were respiratory modulated; of these 53% were inspiratory (I), 36.5% expiratory (E), 6% IE phase spanning and 4.5% EI phase spanning and responsive to airway stimulation. Nearly half (n= 96/200) of the respiratory-modulated units were significantly pulse modulated and 13 were highly modulated with delta(2) values exceeding 0.3. In 10 of these highly modulated units, eta(2) values were greater than 0.3 and all 13 had, at least, a portion of their activity during expiration. We conclude that cardiorespiratory interaction is reciprocal; in addition to respiratory-modulated activity in a subset of neuronal activity patterns controlling the cardiovascular system, pulse-modulated activity exists in a subset of neuronal activity patterns controlling the respiratory system. Thus, cardio-ventilatory coupling apparent in respiratory motor output is evident and, perhaps, derived from the

  10. Activity anorexia: An interplay between basic and applied behavior analysis

    PubMed Central

    Pierce, W. David; Epling, W. Frank; Dews, Peter B.; Estes, William K.; Morse, William H.; Van Orman, Willard; Herrnstein, Richard J.

    1994-01-01

    The relationship between basic research with nonhumans and applied behavior analysis is illustrated by our work on activity anorexia. When rats are fed one meal a day and allowed to run on an activity wheel, they run excessively, stop eating, and die of starvation. Convergent evidence, from several different research areas, indicates that the behavior of these animals and humans who self-starve is functionally similar. A biobehavioral theory of activity anorexia is presented that details the cultural contingencies, behavioral processes, and physiology of anorexia. Diagnostic criteria and a three-stage treatment program for activity-based anorexia are outlined. The animal model permits basic research on anorexia that for practical and ethical reasons cannot be conducted with humans. Thus, basic research can have applied importance. PMID:22478169

  11. Application of activation techniques to biological analysis. [813 references

    SciTech Connect

    Bowen, H.J.M.

    1981-12-01

    Applications of activation analysis in the biological sciences are reviewed for the period of 1970 to 1979. The stages and characteristics of activation analysis are described, and its advantages and disadvantages enumerated. Most applications involve activation by thermal neutrons followed by either radiochemical or instrumental determination. Relatively little use has been made of activation by fast neutrons, photons, or charged particles. In vivo analyses are included, but those based on prompt gamma or x-ray emission are not. Major applications include studies of reference materials, and the elemental analysis of plants, marine biota, animal and human tissues, diets, and excreta. Relatively little use of it has been made in biochemistry, microbiology, and entomology, but it has become important in toxicology and environmental science. The elements most often determined are Ag, As, Au, Br, Ca, Cd, Cl, Co, Cr, Cs, Cu, Fe, Hg, I, K, Mn, Mo, Na, Rb, Sb, Sc, Se, and Zn, while few or no determinations of B, Be, Bi, Ga, Gd, Ge, H, In, Ir, Li, Nd, Os, Pd, Pr, Pt, Re, Rh, Ru, Te, Tl, or Y have been made in biological materials.

  12. Differential Immunohistochemical Profiles for Distinguishing Prostate Carcinoma and Urothelial Carcinoma

    PubMed Central

    Oh, Woo Jin; Chung, Arthur Minwoo; Kim, Jee Soon; Han, Ji Heun; Hong, Sung Hoo; Lee, Ji Yeol; Choi, Yeong Jin

    2016-01-01

    Background The pathologic distinction between high-grade prostate adenocarcinoma (PAC) involving the urinary bladder and high-grade urothelial carcinoma (UC) infiltrating the prostate can be difficult. However, making this distinction is clinically important because of the different treatment modalities for these two entities. Methods A total of 249 patient cases (PAC, 111 cases; UC, 138 cases) collected between June 1995 and July 2009 at Seoul St. Mary’s Hospital were studied. An immunohistochemical evaluation of prostatic markers (prostate-specific antigen [PSA], prostate-specific membrane antigen [PSMA], prostate acid phosphatase [PAP], P501s, NKX3.1, and α-methylacyl coenzyme A racemase [AMACR]) and urothelial markers (CK34βE12, p63, thrombomodulin, S100P, and GATA binding protein 3 [GATA3]) was performed using tissue microarrays from each tumor. Results The sensitivities of prostatic markers in PAC were 100% for PSA, 83.8% for PSMA, 91.9% for PAP, 93.7% for P501s, 88.3% for NKX 3.1, and 66.7% for AMACR. However, the urothelial markers CK34βE12, p63, thrombomodulin, S100P, and GATA3 were also positive in 1.8%, 0%, 0%, 3.6%, and 0% of PAC, respectively. The sensitivities of urothelial markers in UC were 75.4% for CK34βE12, 73.9% for p63, 45.7% for thrombomodulin, 22.5% for S100P, and 84.8% for GATA3. Conversely, the prostatic markers PSA, PSMA, PAP, P501s, NKX3.1, and AMACR were also positive in 9.4%, 0.7%, 18.8%, 0.7%, 0%, and 8.7% of UCs, respectively. Conclusions Prostatic and urothelial markers, including PSA, NKX3.1, p63, thrombomodulin, and GATA3 are very useful for differentiating PAC from UC. The optimal combination of prostatic and urothelial markers could improve the ability to differentiate PAC from UC pathologically. PMID:27498545

  13. Multifractal detrended fluctuation analysis of optogenetic modulation of neural activity

    NASA Astrophysics Data System (ADS)

    Kumar, S.; Gu, L.; Ghosh, N.; Mohanty, S. K.

    2013-02-01

    Here, we introduce a computational procedure to examine whether optogenetically activated neuronal firing recordings could be characterized as multifractal series. Optogenetics is emerging as a valuable experimental tool and a promising approach for studying a variety of neurological disorders in animal models. The spiking patterns from cortical region of the brain of optogenetically-stimulated transgenic mice were analyzed using a sophisticated fluctuation analysis method known as multifractal detrended fluctuation analysis (MFDFA). We observed that the optogenetically-stimulated neural firings are consistent with a multifractal process. Further, we used MFDFA to monitor the effect of chemically induced pain (formalin injection) and optogenetic treatment used to relieve the pain. In this case, dramatic changes in parameters characterizing a multifractal series were observed. Both the generalized Hurst exponent and width of singularity spectrum effectively differentiates the neural activities during control and pain induction phases. The quantitative nature of the analysis equips us with better measures to quantify pain. Further, it provided a measure for effectiveness of the optogenetic stimulation in inhibiting pain. MFDFA-analysis of spiking data from other deep regions of the brain also turned out to be multifractal in nature, with subtle differences in the parameters during pain-induction by formalin injection and inhibition by optogenetic stimulation. Characterization of neuronal firing patterns using MFDFA will lead to better understanding of neuronal response to optogenetic activation and overall circuitry involved in the process.

  14. neutron activation analysis using thermochromatography. III. analysis of samples of biological origin

    SciTech Connect

    Sattarov, G.; Davydov, A.V.; Khamatov, S.; Kist, A.A.

    1986-07-01

    The use of gas thermochromatography (GTC) in the radioactivation analysis of biological materials is discussed. A group separation of a number of highly volatile elements from sodium and bromine radionuclides has been achieved. The limit of detection of the elements by INAA and neutron activation analysis was estimated using GTC. The advantages of the procedure and the analytical parameters are discussed.

  15. Intrapulmonary solitary fibrous tumors: clinicopathologic and immunohistochemical study of 24 cases.

    PubMed

    Rao, Nagarjun; Colby, Thomas V; Falconieri, Giovanni; Cohen, Hector; Moran, Cesar A; Suster, Saul

    2013-02-01

    Solitary fibrous tumor (SFT) is a ubiquitous neoplasm that arises most commonly from the pleura. SFT arising within lung parenchyma (intrapulmonary SFT) has been rarely reported and is therefore not well recognized. We present a clinicopathologic and immunohistochemical study of 24 cases of primary intrapulmonary SFT. Patients' ages ranged from 44 to 83 years (mean, 58 y). None of the patients had evidence or history of a similar tumor elsewhere. Tumor size ranged from 2.3 to 22 cm (mean, 8.5 cm). On the basis of the degree of cytologic atypia, cellularity, mitotic activity, and areas of necrosis, the lesions were divided into low-grade, intermediate-grade, and high-grade histology. Twenty-one tumors showed the conventional features of SFT of low-grade histology (<5 mitoses per 10 high-power fields), with alternating bands of rope-like collagen flanked by a bland-appearing spindle cell proliferation. Hemangiopericytic, angiofibromatous, and a neural-like plexiform growth pattern were also observed. Five of 21 cases showed an "adenofibromatous" appearance imparted by entrapped normal airspaces at the advancing edges of the lesion. One intermediate-grade tumor showed overall increased cellularity with plump, pleomorphic nuclei, 5 to 10 mitoses per 10 high-power fields, and focal areas of classic SFT. Two cases showed high-grade features at initial presentation, with areas resembling a pleomorphic high-grade sarcoma admixed with foci of conventional, low-grade SFT. Immunohistochemical staining analyses performed in 13 cases showed positivity of the tumor cells for CD34, bcl-2, and CD99 in the majority of cases tested. Clinical follow-up was available in 18 patients, with long-term follow-up (>5 y) in 6. Fourteen (14/18) patients were alive and well without evidence of disease 1 month to 14 years after initial diagnosis. Three patients died of their tumors after 4, 5, and 7 years; in 2 of them the initial tumor was of low-grade histology, but the recurrence

  16. Myoepithelial tumors of salivary glands: a clinicopathologic, immunohistochemical, ultrastructural, and flow-cytometric study.

    PubMed

    Alós, L; Cardesa, A; Bombí, J A; Mallofré, C; Cuchi, A; Traserra, J

    1996-05-01

    Myoepitheliomas of the salivary glands remain a controversial entity. To contribute to the knowledge of this entity, 16 myoepithelial tumors of the salivary glands were studied: 12 benign myoepitheliomas (BME) and 4 malignant myoepitheliomas (MME). The clinical and the histologic findings of each case were studied Immunohistochemistry and flow-cytometry analysis were performed from the paraffin-embedded material in 15 cases. An electron-microscopy study was performed in 8 cases. The myoepithelial tumors affected patients of both sexes equally. The mean age of the patients with BME was 54 years, and the mean age of patients with MME was 62 years. Eight cases of BME originated in the parotid gland and 4 cases originated in the minor salivary glands. All the MME developed from a benign preexistent tumor: two developed from a pleomorphic adenoma in the parotid gland, and the other two MME developed in the minor salivary gland from a BME. The myoepithelial tumors were composed of epithelioid, plasmacytoid, spindle, or clear cell types, and they showed a solid or a myxoid pattern of growth. Immunohistochemical studies revealed marked and diffuse positivity to cytokeratins, vimentin, and S-100 protein in all cases. Glial fibrillary acidic protein was positive in 8 cases (53%), and muscle-specific actin and smooth-muscle actin were positive in only 3 cases (20%); they were all cases of BME. Desmin was negative in all tumors. Ultrastructural studies showed the presence of basal membrane, tight junctions, intermediate filaments, and microvilli as well as actin-like filaments lacking focal densities in all cases. But actin-like filaments with focal densities were not identified. Flow cytometry determined that all BME were diploid with a mean proliferative index of 7.73%. Two of the MME were diploid and the other two MME were aneuploid. The mean proliferative index of MME was 11.93%. In conclusion, BME and MME originated in major and minor salivary glands can display different

  17. Finite element analysis of lightweight active primary mirror

    NASA Astrophysics Data System (ADS)

    Lu, Wei Xin; Guan, Chun Lin; Rao, Chang Hui

    2012-09-01

    With the increasing requirement on spatial resolution to achieve ideal performance in space-based optical imaging system, there is a need to enlarge primary apertures. However, primary mirrors of such systems cannot maintain its optical tolerances across the mirror surface after sending to space, because of gravity change and varying ambient temperature. It necessitates active optics technology of primary mirror surface correction. Since mass-to-orbit is expensive and limited, lightweight primary mirror is needed. The paper investigates a lightweight, active primary mirror. This primary mirror structure includes lightweight face sheet and substrate with surface-parallel actuators embedded in the recess of web support ribs. Finite element models of lightweight, active primary mirror structures with different structural parameters are established and simulated. Using the response function matrixes acquired from finite element analysis, the fitting errors for Zernike polynomials are computed by MATLAB. Correctability comparisons of lightweight, active primary mirror structures with different parameters are carried out. To get best correctability, the mirrors should have small recess depth, high and thin ribs, thick face sheets and long actuators. The structural analysis result will be valuable for the design of lightweight, active primary mirror.

  18. Task Analysis of Shuttle Entry and Landing Activities

    NASA Technical Reports Server (NTRS)

    Holland, Albert W.; Vanderark, Stephen T.

    1993-01-01

    The Task Analysis of Shuttle Entry and Landing (E/L) Activities documents all tasks required to land the Orbiter following an STS mission. In addition to analysis of tasks performed, task conditions are described, including estimated time for completion, altitude, relative velocity, normal and lateral acceleration, location of controls operated or monitored, and level of g's experienced. This analysis precedes further investigations into potential effects of zero g on piloting capabilities for landing the Orbiter following long-duration missions. This includes, but is not limited to, researching the effects of extended duration missions on piloting capabilities. Four primary constraints of the analysis must be clarified: (1) the analysis depicts E/L in a static manner--the actual process is dynamic; (2) the task analysis was limited to a paper analysis, since it was not feasible to conduct research in the actual setting (i.e., observing or filming duration an actual E/L); (3) the tasks included are those required for E/L during nominal, daylight conditions; and (4) certain E/L tasks will vary according to the flying style of each commander.

  19. Analysis of alternatives for immobilized low activity waste disposal

    SciTech Connect

    Burbank, D.A.

    1997-10-28

    This report presents a study of alternative disposal system architectures and implementation strategies to provide onsite near-surface disposal capacity to receive the immobilized low-activity waste produced by the private vendors. The analysis shows that a flexible unit strategy that provides a suite of design solutions tailored to the characteristics of the immobilized low-activity waste will provide a disposal system that best meets the program goals of reducing the environmental, health, and safety impacts; meeting the schedule milestones; and minimizing the life-cycle cost of the program.

  20. Multi-scale statistical analysis of coronal solar activity

    DOE PAGES

    Gamborino, Diana; del-Castillo-Negrete, Diego; Martinell, Julio J.

    2016-07-08

    Multi-filter images from the solar corona are used to obtain temperature maps that are analyzed using techniques based on proper orthogonal decomposition (POD) in order to extract dynamical and structural information at various scales. Exploring active regions before and after a solar flare and comparing them with quiet regions, we show that the multi-scale behavior presents distinct statistical properties for each case that can be used to characterize the level of activity in a region. Information about the nature of heat transport is also to be extracted from the analysis.

  1. Analysis of exposure due to work on activated components

    SciTech Connect

    Cossairt, J.D.

    1987-09-01

    In this brief note the author summarized analysis of the exposure incurred in various maintenance jobs involving activated accelerator and beam line components at Fermilab. A tabulation was made of parameters associated with each job. Included are rather terse descriptions of the various tasks. The author presented various plots of the quantities in the table. All exposure rates are mR/hr while all exposures accumulated are mR. The exposure rates were generally measured at the Fermilab standard one foot distance from the activated component. Accumulated exposures are taken from the self-reading pocket dosimeter records maintained by the radiation control technicians.

  2. Dentin matrix protein 1 and dentin sialophosphoprotein in human sound and carious teeth: an immunohistochemical and colorimetric assay.

    PubMed

    Martini, D; Trirè, A; Breschi, L; Mazzoni, A; Teti, G; Falconi, M; Ruggeri, A

    2013-10-29

    Dentin matrix protein 1 (DMP1) and dentin sialophosphoprotein (DSPP) are extracellular matrix proteins produced by odontoblasts involved in the dentin mineralization. The aim this study was to compare the distribution of DMP1 and DSPP in human sound dentin vs human sclerotic dentin. Sixteen sound and sixteen carious human molars were selected, fixed in paraformaldehyde and processed for immunohistochemical detection of DMP1 and DSPP by means of light microscopy, transmission electron microscopy (TEM) and high-resolution field emission in-lens scanning electron microscopy (FEI-SEM). Specimens were submitted to a pre-embedding or a post-embedding immunolabeling technique using primary antibodies anti DMP1 and anti-DSPP and gold-conjugated secondary antibodies. Other samples were processed for the detection of DMP1 and DSPP levels. Dentin from these samples was mechanically fractured to powder, then a protein extraction and a protein level detection assay were performed. DMP1 and DSPP were more abundant in carious than in sound samples. Immunohistochemical analyses in sclerotic dentin disclosed a high expression of DMP1 and DSPP inside the tubules, suggesting an active biomineralization of dentin by odontoblasts. Furthermore, the detection of small amounts of these proteins inside the tubules far from the carious lesion, as shown in the present study, is consistent with the hypothesis of a preventive defense of all dentin after a noxious stimulus has undermined the tooth.

  3. Geographic tongue and psoriasis: clinical, histopathological, immunohistochemical and genetic correlation - a literature review.

    PubMed

    Picciani, Bruna Lavinas Sayed; Domingos, Tábata Alves; Teixeira-Souza, Thays; Santos, Vanessa de Carla Batista Dos; Gonzaga, Heron Fernando de Sousa; Cardoso-Oliveira, Juliana; Gripp, Alexandre Carlos; Dias, Eliane Pedra; Carneiro, Sueli

    2016-01-01

    Geographic tongue is a chronic, inflammatory, and immune-mediated oral lesion of unknown etiology. It is characterized by serpiginous white areas around the atrophic mucosa, which alternation between activity, remission and reactivation at various locations gave the names benign migratory glossitis and wandering rash of the tongue. Psoriasis is a chronic inflammatory disease with frequent cutaneous involvement and an immunogenetic basis of great importance in clinical practice. The association between geographic tongue and psoriasis has been demonstrated in various studies, based on observation of its fundamental lesions, microscopic similarity between the two conditions and the presence of a common genetic marker, human leukocyte antigen (HLA) HLA-C*06. The difficulty however in accepting the diagnosis of geographic tongue as oral psoriasis is the fact that not all patients with geographic tongue present psoriasis. Some authors believe that the prevalence of geographic tongue would be much greater if psoriatic patients underwent thorough oral examination. This study aimed to develop a literature review performed between 1980 and 2014, in which consultation of theses, dissertations and selected scientific articles were conducted through search in Scielo and Bireme databases, from Medline and Lilacs sources, relating the common characteristics between geographic tongue and psoriasis. We observed that the frequency of oral lesions is relatively common, but to establish a correct diagnosis of oral psoriasis, immunohistochemical and genetic histopathological analyzes are necessary, thus highlighting the importance of oral examination in psoriatic patients and cutaneous examination in patients with geographic tongue. PMID:27579734

  4. Geographic tongue and psoriasis: clinical, histopathological, immunohistochemical and genetic correlation - a literature review.

    PubMed

    Picciani, Bruna Lavinas Sayed; Domingos, Tábata Alves; Teixeira-Souza, Thays; Santos, Vanessa de Carla Batista Dos; Gonzaga, Heron Fernando de Sousa; Cardoso-Oliveira, Juliana; Gripp, Alexandre Carlos; Dias, Eliane Pedra; Carneiro, Sueli

    2016-01-01

    Geographic tongue is a chronic, inflammatory, and immune-mediated oral lesion of unknown etiology. It is characterized by serpiginous white areas around the atrophic mucosa, which alternation between activity, remission and reactivation at various locations gave the names benign migratory glossitis and wandering rash of the tongue. Psoriasis is a chronic inflammatory disease with frequent cutaneous involvement and an immunogenetic basis of great importance in clinical practice. The association between geographic tongue and psoriasis has been demonstrated in various studies, based on observation of its fundamental lesions, microscopic similarity between the two conditions and the presence of a common genetic marker, human leukocyte antigen (HLA) HLA-C*06. The difficulty however in accepting the diagnosis of geographic tongue as oral psoriasis is the fact that not all patients with geographic tongue present psoriasis. Some authors believe that the prevalence of geographic tongue would be much greater if psoriatic patients underwent thorough oral examination. This study aimed to develop a literature review performed between 1980 and 2014, in which consultation of theses, dissertations and selected scientific articles were conducted through search in Scielo and Bireme databases, from Medline and Lilacs sources, relating the common characteristics between geographic tongue and psoriasis. We observed that the frequency of oral lesions is relatively common, but to establish a correct diagnosis of oral psoriasis, immunohistochemical and genetic histopathological analyzes are necessary, thus highlighting the importance of oral examination in psoriatic patients and cutaneous examination in patients with geographic tongue.

  5. Immunohistochemical localization of androgen and progesterone receptors in the uterus of the camel (Camelus dromedarius).

    PubMed

    Emam, Mahmoud Abdelghaffar

    2014-10-01

    Ten adult, cyclic female camels (Camelus dromedarius) were used to describe the distribution of androgen (AR) and progesterone (PR) receptors in the uterus using immunohistochemistry. Both AR and PR were distributed throughout the different compartments of the uterus with nuclear staining for AR and PR seen in the cells of epithelia (luminal and glandular), stroma and myometrial smooth muscles. AR immunostaining was not uniform in distribution and intensity; the surface epithelium and the glandular epithelium in the adluminal region of the endometrium showed lower AR immunoreactivity than other compartments of the uterus. PR immunostaining showed uniformity in both distribution and intensity strong PR immunostaining intensity in almost all cells of the different uterine compartments. The intensity and distribution of PR immunostaining in epithelia of lumen and glands in the adluminal regions of endometrium was higher (P<0.05) than that of AR. In conclusion, immunohistochemical localization of AR and PR in the uterus of the cyclic dromedary camel indicates the important roles of androgen and progesterone in controlling the uterine activity during the follicular phase.

  6. Solitary fibrous tumor: histological and immunohistochemical spectrum of benign and malignant variants presenting at different sites.

    PubMed

    Hanau, C A; Miettinen, M

    1995-04-01

    Twenty-nine tumors (from 26 patients, including two with recurrent disease) diagnosed as solitary fibrous tumor (SFT) of the pleura (n = 23), mediastinum (n = 4), abdominal cavity (n = 1), and parotid gland (n = 1) were studied immunohistochemically. Three histologically malignant tumors showed areas of high cellularity and mitotic activity (more than 4 mitoses/10 high-power fields) with features resembling malignant fibrous histiocytoma, malignant hemangiopericytoma, or fibrosarcoma, together with areas typical of benign solitary fibrous tumor. Formaldehyde-fixed, paraffin-embedded tissues and avidin-biotin-complex immunostaining were used. All of the tumors showed vimentin positivity and did not stain for cytokeratin, glial fibrillary acidic protein, or muscle cell markers, except for focal desmin reactivity in seven tumors, mostly seen in frozen sections, and focal keratin reactivity in one histologically malignant tumor. The neoplastic cells were positive for CD34 and negative for CD31; these patterns also were seen in the three histologically malignant cases. In nine of the cases acetone-fixed frozen sections showed variable focal positivity for neurofilament proteins of 68 kd. We conclude that SFT is a neoplasm of fibroblasts/primitive mesenchymal cells with features of multidirectional differentiation. We also report the finding of a novel site for SFT, the parotid gland.

  7. [T lymphocyte populations of the intestinal mucosa in celiac disease in children. Immunohistochemical study].

    PubMed

    Olives, J P; Voigt, J J; al Saati, T; Nonnenmacher, L; Brousset, P; Delsol, G; Ghisolfi, J

    1990-01-01

    In order to study the distribution of lymphocyte subpopulations in a pathologic intestinal mucosa, the authors, instead of using the classic method by counting the number of lymphocytes, present an original method permitting the exploitation of quantified data from labelled surface cells by texture analyser coupled with a computerized system. We investigated 25 children presenting with chronic diarrhea and villous atrophy and 5 control subjects. Fifteen of the 25 children had celiac disease (10 active with total villous atrophy and 5, celiac disease in remission with healing mucosa), 5 cow's milk protein intolerance with total or partial villous atrophy and 5, chronic diarrhea with partial villous atrophy. Immunohistochemical study with monoclonal antibodies was carried out on frozen sections using a three-step immunoperoxidase technique. Compared with the 5 controls, patients with food intolerance (celiac disease and cow's milk protein intolerance) showed a significant increase of T suppressor lymphocytes (p less than 0.01 and p less than 0.05) in the epithelium, whereas there were more T helper lymphocytes in the lamina propria (p less than 0.05 and p less than 0.01). Non-treated celiac disease was distinguished from treated celiac disease by a marked increase in intra-epithelial T cytotoxic-suppressors. These results suggest that T cytotoxic-suppressors may be the mediators of the lesions observed in celiac disease. PMID:2179007

  8. Immunohistochemical Expression of GLUT-1 and HIF-1α in Tooth Germ, Ameloblastoma, and Ameloblastic Carcinoma.

    PubMed

    Sánchez-Romero, Celeste; Bologna-Molina, Ronell; Mosqueda-Taylor, Adalberto; Paes de Almeida, Oslei

    2016-08-01

    Hypoxia-inducible factor-1α (HIF-1α) promotes proteins that enable cell survival during hypoxia, such as glucose transporter 1 (GLUT-1). Their coexpression has been associated with aggressiveness in malignancies and has not been studied in odontogenic tumors. Immunohistochemical expression of HIF-1α and GLUT-1 was analyzed in 13 tooth germs (TGs), 55 ameloblastomas (AMs), and 3 ameloblastic carcinomas (ACs). HIF-1α was negative in all TGs, and just 1 case of AM and 1 of AC had nuclear positivity. GLUT-1 expressed in ameloblastic cells of all TGs, AMs, and ACs, with an increasing intensity, respectively, and was significantly higher in solid AM than in unicystic AM (P = .041). Absence of nuclear HIF-1α in TGs and most AMs suggest that GLUT-1 may be induced by alternative pathways to hypoxia. However, in ACs, HIF-1α may be activated; however, to confirm this, additional cases are needed. GLUT-1 overexpression could be related to aggressiveness in AMs and ACs and must represent a normal metabolite in TGs. PMID:27020375

  9. A reappraisal of macrophage polarization in glioblastoma: Histopathological and immunohistochemical findings and review of the literature.

    PubMed

    Mignogna, Chiara; Signorelli, Francesco; Vismara, Marco Flavio Michele; Zeppa, Pio; Camastra, Caterina; Barni, Tullio; Donato, Giuseppe; Di Vito, Anna

    2016-06-01

    The survival rate in glioblastoma multiforme patients has scarcely improved in the last decades; however, many new therapeutic strategies have been theorized or developed for these neoplasias. Recently, the inverse correlation observed between patient prognosis and tumor-associated macrophages (TAMs) density in solid tumors has encouraged the development of anti-tumor strategies aiming to target TAMs. As expected, TAMs polarization is influenced by both macrophage localization and tumor microenvironment signals, resulting in a more complex scenario than the simple M1/M2 activation status. Macrophage polarization in glioblastoma has not yet been fully elucidated, and most results have been obtained in experimental non-human settings, with some apparent contradiction. The authors performed a histopathological and immunohistochemical study of 37 cases of glioblastoma in order to characterize the M1 and M2 macrophage populations within TAMs. A high prevalence of CD163+ M2-polarized macrophages was detected in this cohort, whereas iNOS+ macrophages were rarely found. The down-regulation of CD68 expression in microglia/macrophage infiltrating glioblastomas is also reported for the first time. Such a finding is associated with a specific location of TAMs within the lesion, as confirmed by the fact that CD68 staining was lower than CD163, mainly in perivascular areas. The authors discuss the recent literature about the global scenario of macrophage plasticity and polarization in glioblastoma, and suggest some pivotal points for therapeutic applications.

  10. Sarcoglycan complex in masseter and sternocleidomastoid muscles of baboons: an immunohistochemical study.

    PubMed

    Cutroneo, G; Centofanti, A; Speciale, F; Rizzo, G; Favaloro, A; Santoro, G; Bruschetta, D; Milardi, D; Micali, A; Di Mauro, D; Vermiglio, G; Anastasi, G; Trimarchi, F

    2015-01-01

    The sarcoglycan complex consists of a group of single-pass transmembrane glycoproteins that are essential to maintain the integrity of muscle membranes. Any mutation in each sarcoglycan gene causes a series of recessive autosomal dystrophin-positive muscular dystrophies. Negative fibres for sarcoglycans have never been found in healthy humans and animals. In this study, we have investigated whether the social ranking has an influence on the expression of sarcoglycans in the skeletal muscles of healthy baboons. Biopsies of masseter and sternocleidomastoid muscles were processed for confocal immunohistochemical detection of sarcoglycans. Our findings showed that baboons from different social rankings exhibited different sarcoglycan expression profiles. While in dominant baboons almost all muscles were stained for sarcoglycans, only 55% of muscle fibres showed a significant staining. This different expression pattern is likely to be due to the living conditions of these primates. Sarcoglycans which play a key role in muscle activity by controlling contractile forces may influence the phenotype of muscle fibres, thus determining an adaptation to functional conditions. We hypothesize that this intraspecies variation reflects an epigenetic modification of the muscular protein network that allows baboons to adapt progressively to a different social status.

  11. Sarcoglycan Complex in Masseter and Sternocleidomastoid Muscles of Baboons: An Immunohistochemical Study

    PubMed Central

    Cutroneo, G.; Centofanti, A.; Speciale, F.; Rizzo, G.; Favaloro, A.; Santoro, G.; Bruschetta, D.; Milardi, D.; Micali, A.; Di Mauro, D.; Vermiglio, G.; Anastasi, G.; Trimarchi, F.

    2015-01-01

    The sarcoglycan complex consists of a group of single-pass transmembrane glycoproteins that are essential to maintain the integrity of muscle membranes. Any mutation in each sarcoglycan gene causes a series of recessive autosomal dystrophin-positive muscular dystrophies. Negative fibres for sarcoglycans have never been found in healthy humans and animals. In this study, we have investigated whether the social ranking has an influence on the expression of sarcoglycans in the skeletal muscles of healthy baboons. Biopsies of masseter and sternocleidomastoid muscles were processed for confocal immunohistochemical detection of sarcoglycans. Our findings showed that baboons from different social rankings exhibited different sarcoglycan expression profiles. While in dominant baboons almost all muscles were stained for sarcoglycans, only 55% of muscle fibres showed a significant staining. This different expression pattern is likely to be due to the living conditions of these primates. Sarcoglycans which play a key role in muscle activity by controlling contractile forces may influence the phenotype of muscle fibres, thus determining an adaptation to functional conditions. We hypothesize that this intraspecies variation reflects an epigenetic modification of the muscular protein network that allows baboons to adapt progressively to a different social status. PMID:26150161

  12. Localization of molecules with restricted patterns of expression in morphogenesis: an immunohistochemical approach.

    PubMed

    Thibodeau, A; Duchaine, J; Simard, J L; Vincent, M

    1989-06-01

    In a search for molecules with restricted patterns of expression during development, monoclonal antibodies were raised against different transitory structures of the chick embryo. Mice were immunized with cell suspensions from lightly homogenized embryonic tissues explanted from morphogenetically active regions. A convenient immunohistochemical assay was used to screen the hybridoma supernatants on a large scale. It relied on the use of poly(ethylene glycol) as embedding medium. Its water miscibility allowed, in a one-step incubation with antibody-containing supernatants, the dewaxing and rehydration of the tissue sections as well as antibody binding. We report here the usefulness of this approach in selecting monoclonals with unique patterns of immunoreactivity. In this study, cephalic neural crest cells in early or late phase of migration, together with their surrounding tissues, were used as immunogens. The monoclonal antibodies obtained have been classified into regional, cell-lineage, cell-cycle or extracellular material-associated markers. The information provided by the direct visualization of the immunoreactivity of the various monoclonal antibodies on tissue sections, as early as the first round of screening, allows rapid determination of the subsequent strategy to be followed for further characterization of the individual markers.

  13. Geographic tongue and psoriasis: clinical, histopathological, immunohistochemical and genetic correlation - a literature review*

    PubMed Central

    Picciani, Bruna Lavinas Sayed; Domingos, Tábata Alves; Teixeira-Souza, Thays; dos Santos, Vanessa de Carla Batista; Gonzaga, Heron Fernando de Sousa; Cardoso-Oliveira, Juliana; Gripp, Alexandre Carlos; Dias, Eliane Pedra; Carneiro, Sueli

    2016-01-01

    Geographic tongue is a chronic, inflammatory, and immune-mediated oral lesion of unknown etiology. It is characterized by serpiginous white areas around the atrophic mucosa, which alternation between activity, remission and reactivation at various locations gave the names benign migratory glossitis and wandering rash of the tongue. Psoriasis is a chronic inflammatory disease with frequent cutaneous involvement and an immunogenetic basis of great importance in clinical practice. The association between geographic tongue and psoriasis has been demonstrated in various studies, based on observation of its fundamental lesions, microscopic similarity between the two conditions and the presence of a common genetic marker, human leukocyte antigen (HLA) HLA-C*06. The difficulty however in accepting the diagnosis of geographic tongue as oral psoriasis is the fact that not all patients with geographic tongue present psoriasis. Some authors believe that the prevalence of geographic tongue would be much greater if psoriatic patients underwent thorough oral examination. This study aimed to develop a literature review performed between 1980 and 2014, in which consultation of theses, dissertations and selected scientific articles were conducted through search in Scielo and Bireme databases, from Medline and Lilacs sources, relating the common characteristics between geographic tongue and psoriasis. We observed that the frequency of oral lesions is relatively common, but to establish a correct diagnosis of oral psoriasis, immunohistochemical and genetic histopathological analyzes are necessary, thus highlighting the importance of oral examination in psoriatic patients and cutaneous examination in patients with geographic tongue. PMID:27579734

  14. Immunohistochemical Studies on Galectin Expression in Colectomised Patients with Ulcerative Colitis

    PubMed Central

    Block, Mattias; Mölne, Johan; Leffler, Hakon; Börjesson, Lars; Breimer, Michael E.

    2016-01-01

    Introduction. The aetiology and pathogenesis of ulcerative colitis (UC) are essentially unknown. Galectins are carbohydrate-binding lectins involved in a large number of physiological and pathophysiological processes. Little is known about the role of galectins in human UC. In this immunohistochemical exploratory study, both epithelial and inflammatory cell galectin expression were studied in patients with a thoroughly documented clinical history and were correlated with inflammatory activity. Material and Methods. Surgical whole intestinal wall colon specimens from UC patients (n = 22) and controls (n = 10) were studied. Clinical history, pharmacological treatment, and modified Mayo-score were recorded. Tissue inflammation was graded, and sections were stained with antibodies recognizing galectin-1, galectin-2, galectin-3, and galectin-4. Results. Galectin-1 was undetectable in normal and UC colonic epithelium, while galectin-2, galectin-3, and galectin-4 were strongly expressed. A tendency towards diminished epithelial expression with increased inflammatory grade for galectin-2, galectin-3, and galectin-4 was also found. In the inflammatory cells, a strong expression of galectin-2 and a weak expression of galectin-3 were seen. No clear-cut correlation between epithelial galectin expression and severity of the disease was found. Conclusion. Galectin expression in patients with UC seems to be more dependent on disease focality and individual variation than on degree of tissue inflammation. PMID:26885508

  15. Sebaceous lymphadenoma of the thymus: A clinicopathologic and immunohistochemical study of 2 cases.

    PubMed

    Weissferdt, Annikka; Kalhor, Neda; Moran, Cesar A

    2016-10-01

    Two cases of primary sebaceous lymphadenoma of the thymus are presented. The patients were a man and a woman 58 and 77 years old, respectively. The female patient had a history of breast carcinoma and on follow-up was identified to have an anterior mediastinal mass; the male patient did not have any history of malignancy, and the tumor was discovered during a chest radiographic evaluation when the patient presented with symptoms of fatigue, chest pain, and dyspnea. Histologically, both lesions were characterized by the presence of solid-cystic epithelial islands in a prominent lymphocytic background. The epithelial islands were haphazardly distributed in the form of small tubular structures with focal keratinization and groups of epithelial cells with clear cytoplasm, round nuclei and lack of mitotic activity in keeping with sebaceous cells. The presence of germinal centers in the lymphoid background was seen in both cases. Immunohistochemically, the epithelial component was positive for cytokeratin 8 (CAM5.2), cytokeratin 5/6, and for adipophilin in the sebaceous component. B- and T-cell markers were positive in the lymphoid component. Clinical follow-up in both patients showed that the 2 patients were well and alive 3 years after diagnosis. The cases herein presented expand the spectrum of salivary gland-type tumors in the mediastinum and raise awareness of lesions which are easily confused with other more common thymic tumors that have different prognosis and treatment implications. PMID:27374279

  16. Overlapping Morphologic and Immunohistochemical Features of Hashimoto Thyroiditis and IgG4-Related Thyroid Disease.

    PubMed

    Raess, Philipp W; Habashi, Arlette; El Rassi, Edward; Milas, Mira; Sauer, David A; Troxell, Megan L

    2015-05-01

    Immunoglobulin G4-related disease (IgG4-RD) is an emerging clinicopathologic entity characterized by both IgG4+ plasma cell infiltration and fibrosis in one or more organs, prototypically pancreas or salivary/lacrimal glands. IgG4-RD in the thyroid (IgG4-RTD) is an area of active study, and the relationship between IgG4-RTD and Hashimoto thyroiditis is not fully delineated due to their overlapping histologic features. Retrospective review was performed of all thyroidectomy cases demonstrating lymphocytic inflammation at a single institution over a 4-year period. Approximately half (23/38) of patients had a clinical diagnosis of Hashimoto thyroiditis (HT). Nine of the 38 patients had increased absolute and relative numbers of IgG4+ plasma cells. Patients with a clinical diagnosis of HT had increased lymphoplasmacytic inflammation, but the relative proportion of IgG4+ plasma cells was not increased compared to patients without HT. There was no correlation between IgG4 levels and the amount of fibrosis in patients with or without HT. Patients identified as having the fibrosing variant of HT were not more likely to have increased levels of IgG4+ plasma cells than those without. There is significant morphologic and immunohistochemical overlap between HT and IgG4-RTD. Future studies to identify specific characteristics of IgG4-RTD involving the thyroid are necessary to accurately define this entity.

  17. Immunohistochemical localization of androgen and progesterone receptors in the uterus of the camel (Camelus dromedarius).

    PubMed

    Emam, Mahmoud Abdelghaffar

    2014-10-01

    Ten adult, cyclic female camels (Camelus dromedarius) were used to describe the distribution of androgen (AR) and progesterone (PR) receptors in the uterus using immunohistochemistry. Both AR and PR were distributed throughout the different compartments of the uterus with nuclear staining for AR and PR seen in the cells of epithelia (luminal and glandular), stroma and myometrial smooth muscles. AR immunostaining was not uniform in distribution and intensity; the surface epithelium and the glandular epithelium in the adluminal region of the endometrium showed lower AR immunoreactivity than other compartments of the uterus. PR immunostaining showed uniformity in both distribution and intensity strong PR immunostaining intensity in almost all cells of the different uterine compartments. The intensity and distribution of PR immunostaining in epithelia of lumen and glands in the adluminal regions of endometrium was higher (P<0.05) than that of AR. In conclusion, immunohistochemical localization of AR and PR in the uterus of the cyclic dromedary camel indicates the important roles of androgen and progesterone in controlling the uterine activity during the follicular phase. PMID:25081941

  18. Immunohistochemical studies of the epididymal duct in Egyptian water buffalo (Bubalus bubalis).

    PubMed

    Alkafafy, Mohamed; Elnasharty, Mohamed; Sayed-Ahmed, Ahmed; Abdrabou, Mohamed

    2011-02-01

    Using immunohistochemistry (IHC), this study aimed to evaluate the regional distribution pattern of some biologically active proteins in the epididymis of Egyptian water buffalo and to determine the structural-functional relationships of the different epididymal structures. Wax-embedded sections from different regions of the epididymal duct from adult, clinically healthy, buffalo bulls were used. Primary antibodies against angiotensin converting enzyme (ACE), S-100, galactosyltransferase (GalTase), alpha smooth muscle actin (α-SMA), connexin 43 (Cx43) and vascular endothelial growth factor (VEGF) were used for immunohistochemical studies. The results showed that, in addition to the well-known principal and basal cells, the epididymal epithelium, similar to that of other species, possessed apical cells and intraepithelial leukocytes. IHC showed that, with the exception of VEGF which reacted negatively, all antibodies used displayed variable reactivity in the different epididymal structures. Apical cells expressed a strong reaction with ACE along the entire length of the duct. The principal cells in the caput epididymis exhibited a distinct reactivity with S-100 and GalTase. The peritubular muscular coat displayed a marked immunostaining for α-SMA and for Cx43. In conclusion these findings showed a regional-specific distribution pattern, distinct from that in bovine bulls. Some potential functional capacities, especially absorptive and secretory ones, are discussed in relation to the different epididymal regions.

  19. Carcinoma ex pleomorphic adenoma of the salivary gland: an immunohistochemical study.

    PubMed

    Matsubayashi, Saori; Yoshihara, Toshio

    2007-07-01

    The proliferative activity of the tumor cells and the expression of tumor-associated genes and sex steroid hormone receptors were investigated immunohistochemically in ten cases of carcinoma ex pleomorphic adenoma (Ca-ex-PA) of the salivary glands. These were analyzed in benign and malignant components separately, and then were compared with ten cases of the other malignant tumors [adenocarcinomas, not otherwise specified (ACN) and salivary duct carcinomas (SDC)] and ten cases of pleomorphic adenomas (PA). The results obtained in this study were as follows: (1) malignant component of Ca-ex-PA showed a higher incidence of PCNA and Ki67 than benign component of Ca-ex-PA. A significant difference between benign component of Ca-ex-PA and PA was not observed. (2) A significant difference in the incidence of p53, c-erbB-2, EGFR overexpression was observed only between malignant component of Ca-ex-PA and benign component of Ca-ex-PA. (3) The incidence of PCNA, Ki67, p53, c-erbB-2 overexpression in malignant component of Ca-ex-PA showed the highest data among the four groups. These results suggest that Ca-ex-PA acquired the particular biological behavior in contrast to the other salivary neoplasms in the long-standing process while PA undergoes malignant transformation.

  20. Immunohistochemical localization of T-lymphocyte subsets in the developing lymphoid tissues of the tammar wallaby (Macropus eugenii).

    PubMed

    Duncan, Louise G; Nair, Sham V; Deane, Elizabeth M

    2012-12-01

    Research into marsupial adaptive immunity during ontogeny has been hampered by the lack of antibodies that react to marsupial immunological cell populations. In this study, newly synthesised polyclonal antibodies to the T cell marker, CD8, have been developed and used to investigate the ontogeny and distribution of this T cell population in the tammar wallaby. Immunohistochemical analysis indicated that the distribution of the CD8 lymphocytes in the lymphoid tissues of tammar neonates during the first 144 days of pouch life was similar to that of the eutherian mammals. However, CD8α(+) lymphocytes were observed in the intestines of tammar neonates prior to their first appearance in the cervical thymus, an observation that has not been found in eutherians. A dual labelling immunohistochemical approach was used for the indirect demonstration of CD4 and enabled the simultaneous detection in the tammar wallaby tissues of the two major T-lymphocyte populations, CD4 and CD8 that are associated with adaptive immunity. As in eutherian mammals, CD4(+) cells were the predominant T cell lymphocyte subset observed in the spleen while in the nodal tissues, an age-related decrease in the CD4(+)/CD8(+) ratio was noted. These antibodies provide a new immunological tool to study the role of T cell subsets in marsupial immunity and disease pathogenesis studies. PMID:22929957

  1. Immunohistochemical localization of progesterone receptor isoforms and estrogen receptor alpha in the chicken oviduct magnum during development.

    PubMed

    González-Morán, María Genoveva

    2015-10-01

    In this work, the immunohistochemical expression of progesterone receptor (PR) isoforms and estrogen receptor alpha (ER-α), as well as the histomorphometric changes of the magnum region of the left oviduct from 8-day-old chicken embryos to one-month-old chickens were evaluated. Results indicate evident histological changes in the oviduct magnum during development mainly in the magnum's mucosa. Immunohistochemical analysis showed that the oviduct magnum from 8-day-old chicken embryos to one-day-old chickens did not present any PR isoform, but the oviduct magnum of one-week and one-month-old chickens expressed PR in the nuclei of all cell types. In epithelial cells, PR-B was the only isoform expressed; in muscle and serosa cells, PR-A isoform was the only isoform expressed; and stromal cells expressed both isoforms. The results also demonstrate positive ER-α immunostaining in the nuclei of different cells from embryonic life to later developmental stages of the oviduct magnum. Data indicate that the variations of ER-α or PR expression or dominance of either PR expression is differentially regulated depending on the cell type, the development of the oviduct, and in an age-specific manner. These variations in sex steroids hormone receptors are related with histological changes of the oviduct magnum through development.

  2. Comparative histological and immunohistochemical study of sea star tube feet (Echinodermata, Asteroidea).

    PubMed

    Santos, Romana; Haesaerts, Delphine; Jangoux, Michel; Flammang, Patrick

    2005-03-01

    Adhesion in sea stars is the function of specialized structures, the tube feet or podia, which are the external appendages of the water-vascular system. Adhesive secretions allow asteroid tube feet to perform multiple functions. Indeed, according to the sea star species considered, the tube feet may be involved in locomotion, fixation, or burrowing. Different tube foot shapes usually correspond to this variety of function. In this study, we investigated the variability of the morphology of sea star tube feet as well as the variability of the composition of their adhesive secretions. This second aspect was addressed by a comparative immunohistochemical study using antibodies raised against the adhesive material of the forcipulatid Asterias rubens. The tube feet from 14 sea star species representing five orders and 10 families of the Class Asteroidea were examined. The histological study revealed three main tube foot morphotypes, i.e., knob-ending, simple disc-ending, and reinforced disc-ending. Analysis of the results suggests that tube foot morphology is influenced by species habitat, but within limits imposed by the evolutionary lineage. In immunohistochemistry, on the other hand, the results were very homogeneous. In every species investigated there was a very strong immunolabeling of the adhesive cells, independently of the taxon considered, of the tube foot morphotype or function, or of the species habitat. This indicates that the adhesives in all the species considered are closely related, probably sharing many identical molecules or, at least, many identical epitopes on their constituents.

  3. Immunohistochemical location of serotonin and serotonin 2B receptor in the small intestine of pigs.

    PubMed

    Zhang, Han; Zhang, Tao; Wang, Lei; Teng, Kedao

    2009-01-01

    The distribution of serotonin and serotonin 2B receptor in the small intestines of pigs newborn, 5, 15 and 100 days of age were examined qualitatively and quantitatively by immunohistochemical labeling, microscopic observation and image analysis. The results showed serotonin immunopositive cells distributed diffusely among the epithelial cells of the middle and more basal parts of villi and intestinal glands in all segments of all pigs examined. Serotonin 2B receptor was first localized in the duodenum of 15-day-old pigs, whereas in 100-day-old pigs, serotonin 2B receptor was immunolabeled abundantly in all segments. Serotonin 2B receptor was distributed in the connective tissue of the small intestinal mucosa, lamina propria and in some myenteric neurons. The density of serotonin 2B receptor immunopositive cells in the duodenum of 100-day-old pigs was higher than that of 15-day-old pigs. The density of serotonin 2B receptor immunopositive cells in the duodenum was the highest among the three segments of the 100-day-old pigs. The study indicates that the distribution of serotonin 2B receptor is species different in the pig small intestine and the intensity of serotonin 2B receptor becomes stronger as the small intestine matures.

  4. Tape transfer sectioning of tissue microarrays introduces nonspecific immunohistochemical staining artifacts.

    PubMed

    Catchpoole, D; Mackie, N; McIver, S; Chetcuti, A; Henwood, A; Graf, N; Arbuckle, S

    2011-12-01

    Tissue microarrays place tens to hundreds of formalin fixed, paraffin embedded tissue cores into a paraffin block in a systematic grid pattern that permits their simultaneous evaluation in a single section. The fragmented nature of the tissue cores often makes sectioning of tissue microarrays difficult so that the resulting disks of tissue lose their shape, fracture or fall out of the paraffin section altogether. We have evaluated an alternative sectioning protocol for stabilizing the tissue microarray surface by placing an adhesive tape "window" over the face of the paraffin block prior to sectioning. Once sectioned, the tape/sections are transferred directly onto coated microscope slides, thereby avoiding routine floating of sections on a water bath. After sectioning with either the tape transfer or standard protocols, slides were stained either using hematoxylin and eosin or immunohistochemistry using antibodies to S-100 protein and the tissue specific antigens, keratin (AE1/3) and the leukocyte common antigen CD45. We found that the tape method produced thicker sections that were darker and more densely packed with loss of tissue definition compared to sections prepared using water bath flotation. Quantitative image analysis of immunohistochemical staining demonstrated that the tape method produced a higher incidence of nonspecific staining, which raised the potential for false positive staining.

  5. Quantitative Evaluation of Macrophage Expression Using CD68 in Oral Submucous Fibrosis: An Immunohistochemical Study

    PubMed Central

    Pereira, T; Naik, S; Tamgadge, A

    2015-01-01

    Background: Macrophages are important cells for the innate immunity. Circulating monocytes are attracted to tissues by chemotactic factors and become macrophages under the influence of their microenvironment. Several studies suggested that local and systemic upregulation of fibrogenic cytokines and downregulation of antifibrotic cytokine are central to the pathogenesis of oral submucous fibrosis (OSMF). Currently, there have been no attempts made to elucidate the presence and role of macrophages in OSMF. Aim: Our aim was to study the expression of CD68 in OSMF patients and to investigate the possible correlation of macrophages using CD68 in various histopathological grades of OSMF. Subjects and Methods: A prospective case–control study which included 40 patients was conducted after obtaining informed consent and Ethical Committee clearance. Ten cases were normal control and thirty cases had OSMF. Biopsy was performed and a quantitative study of macrophages was done using CD68 antigen and was immunohistochemically localized. Statistical analysis was carried out using the Statistical Package for Social Sciences (SPSS) 17.0 version (SPSS Inc., Chicago, IL, USA). Results: OSMF was observed in male patients of a younger age group. The macrophage number in the patients of intermediate and advanced stage of OSMF was higher than that of the controls which was statistically significant (P < 0.05). Conclusion: The findings of this study suggest that CD68 plays a vital role in the pathogenesis of OSMF and can be regarded as a useful marker for assessing the progress of the disease. PMID:27057383

  6. Prognostic value of clinical, morphologic, and immunohistochemical factors in patients with bronchiolitis obliterans-organizing pneumonia.

    PubMed

    Pardo, Javier; Panizo, Angel; Sola, Iosu; Queipo, Francisco; Martinez-Peñuela, Ana; Carias, Rafael

    2013-05-01

    Bronchiolitis obliterans-organizing pneumonia (BOOP) is an inflammatory and fibrosing disease involving the distal bronchioles, bronchiolar ducts, and alveoli. We studied 91 patients with BOOP. Univariate analysis was used to relate age, sex, smoking, morphology, and expression of immunohistochemical markers CD68, D2-40, CD31, CD34, collagen IV, collagen III, platelet-derived growth factor receptor, and vascular endothelial growth factor (VEGF) with the response to corticosteroid therapy. Seventy-two patients had idiopathic BOOP and 19 secondary BOOP. The median age of the patients was 59.54 years. Most patients were current or former smokers. All cases had a patchy lesion consisting of small buds of fibromyxoid tissue in small bronchioles, bronchiolar ducts, and alveoli. The buds contained collagen and reticulin fibers, fibroblasts, macrophages, mononuclear inflammatory cells, and vessels in different proportions. We found no morphologic differences between primary and secondary BOOP. Patients younger than 38 years and nonsmokers had a significant good response to corticosteroid therapy. Favorable morphologic predictors were the presence of large bronchial plugs and mild inflammatory reaction (P = .093). By immunohistochemistry, the presence of collagen IV with the absence of collagen III, CD68-positive cells and positive VEGF were associated with a good response to corticosteroid therapy. We conclude that age, smoking, localization, and extension of proliferative intrabronchiolar plugs and positive immunostains for CD68, VEGF, and collagen IV with negative collagen III were useful to predict response to corticosteroid therapy and relapse.

  7. Immunohistochemical expression of p53 and its clinicopathological correlation with modified Anneroth's histological grading system

    PubMed Central

    Dave, Kajal V; Chalishazar, Monali; Dave, Vishal R; Panja, Pritam; Singh, Manisha; Modi, Tapan G

    2016-01-01

    Introduction and Objectives: Oral squamous cell carcinoma (OSCC) is an epithelial neoplasm generally beginning as focal overgrowth of altered stem cells near the basement membrane, moving upward and laterally, replacing the normal epithelium. Histopathological grading has been used for many decades in an attempt to predict the clinical behavior of oral squamous cell carcinoma. In the present study, Forty biopsies were studied for histological grading and p53 expression. The p53 expression was studied in relation to clinical parameters such as age, sex of patient and site of tumors. Relation between histological grade of malignancy and p53 protein expression was analysed. All cases were classified according to Anneroth's histological malignancy grading system (1987). Materials and Methods: 40 cases of OSCC were assessed for clinical parameters, Anneroth's histological grading and immunohistochemically stained with p53 protien. Statistical Analysis: The results obtained were analyzed using Spearman's Co-relation. Observations and Results: The positive expression of p53 was found in 62% of carcinomas studied. Positivity of p53 showed correlation with histological grade of malignancy and with individual parameters like degree of keratinization, nuclear polymorphism, number of mitoses and lymphoplasmacytic infiltration while showed a negative correlation with pattern of invasion. Conclusion: Our study showed a significant correlation between parameters of tumor cell population, lymphoplasmacytic infiltration and p53 expression. A significant association between high grade of malignancy and p53 overexpression and insignificant correlation of p53 with age, sex of the patient and site of the tumor was found. PMID:27194859

  8. Detection of oral squamous cell carcinoma metastasis with cathepsin D: An immunohistochemical approach

    PubMed Central

    Kapoor, Seema; Kaur, Geet Priya; Sikka, Pranav

    2014-01-01

    Background: The lysosomal protease cathepsin D (CD) has been associated with tumor progression in malignant tumors including oral squamous cell carcinoma (OSCC). The purpose of this study was to find out any association between the CD and lymph node metastasis and to study the correlation of CD with various clinicopathological parameters to aid in assessment of its role as a prognostic indicator. Materials and Methods: Immunohistochemical staining was performed on 20 OSCC samples with polyclonal antibody against CD. Positive results indicative of the presence of CD were further analyzed to determine any correlation between the CD and other clinicopathological parameters. Pearson Chi-square analyses, Spearsman correlation coefficient, Mann-Whitney test, Kruskal Wallis test and student t test were used for statistical analysis (P < 0.05). Results: Patients with lymph node metastasis showed statistically significant increase in CD expression (P < 0.01). Increasing tumor size seemed to correlate with increased CD expression (P < 0.05). Conclusion: Based on its association with other clinicopathological variables, CD expression can be used for the assessment of patient survival in cases of OSCC. PMID:24932191

  9. Adult Active Transport in the Netherlands: An Analysis of Its Contribution to Physical Activity Requirements

    PubMed Central

    Fishman, Elliot; Böcker, Lars; Helbich, Marco

    2015-01-01

    Introduction Modern, urban lifestyles have engineered physical activity out of everyday life and this presents a major threat to human health. The Netherlands is a world leader in active travel, particularly cycling, but little research has sought to quantify the cumulative amount of physical activity through everyday walking and cycling. Methods Using data collected as part of the Dutch National Travel Survey (2010 – 2012), this paper determines the degree to which Dutch walking and cycling contributes to meeting minimum level of physical activity of 150 minutes of moderate intensity aerobic activity throughout the week. The sample includes 74,465 individuals who recorded at least some travel on the day surveyed. As physical activity benefits are cumulative, all walking and cycling trips are analysed, including those to and from public transport. These trips are then converted into an established measure of physical activity intensity, known as metabolic equivalents of tasks. Multivariate Tobit regression models were performed on a range of socio-demographic, transport resources, urban form and meteorological characteristics. Results The results reveal that Dutch men and women participate in 24 and 28 minutes of daily physical activity through walking and cycling, which is 41% and 55% more than the minimum recommended level. It should be noted however that some 57% of the entire sample failed to record any walking or cycling, and an investigation of this particular group serves as an important topic of future research. Active transport was positively related with age, income, bicycle ownership, urban density and air temperature. Car ownership had a strong negative relationship with physically active travel. Conclusion The results of this analysis demonstrate the significance of active transport to counter the emerging issue of sedentary lifestyle disease. The Dutch experience provides other countries with a highly relevant case study in the creation of

  10. Diagnostic Application of Absolute Neutron Activation Analysis in Hematology

    SciTech Connect

    Zamboni, C.B.; Oliveira, L.C.; Dalaqua, L. Jr.

    2004-10-03

    The Absolute Neutron Activation Analysis (ANAA) technique was used to determine element concentrations of Cl and Na in blood of healthy group (male and female blood donators), select from Blood Banks at Sao Paulo city, to provide information which can help in diagnosis of patients. This study permitted to perform a discussion about the advantages and limitations of using this nuclear methodology in hematological examinations.

  11. Analyses of Oxyanion Materials by Prompt Gamma Activation Analysis

    SciTech Connect

    Firestone, Richard B; Perry, D.L.; English, G.A.; Firestone, R.B.; Leung, K.-N.; Garabedian, G.; Molnar, G.L.; Revay, Zs.

    2008-03-24

    Prompt gamma activation analysis (PGAA) has been used to analyze metal ion oxyanion materials that have multiple applications, including medicine, materials, catalysts, and electronics. The significance for the need for accurate, highly sensitive analyses for the materials is discussed in the context of quality control of end products containing the parent element in each material. Applications of the analytical data for input to models and theoretical calculations related to the electronic and other properties of the materials are discussed.

  12. Obsidian sources characterized by neutron-activation analysis.

    PubMed

    Gordus, A A; Wright, G A; Griffin, J B

    1968-07-26

    Concentrations of elements such as manganese, scandium, lanthanum, rubidium, samarium, barium, and zirconium in obsidian samples from different flows show ranges of 1000 percent or more, whereas the variation in element content in obsidian samples from a single flow appears to be less than 40 percent. Neutron-activation analysis of these elements, as well as of sodium and iron, provides a means of identifying the geologic source of an archeological artifact of obsidian.

  13. HPTLC Analysis, Antioxidant and Antigout Activity of Indian Plants.

    PubMed

    Nile, Shivraj Hariram; Park, Se Won

    2014-01-01

    The HPTLC analysis, antioxidant, and antigout activity of Asparagus racemosus, Withania somnifera, Vitex negundo, Plumbago zeylanica, Butea monosperma and Tephrosia purpurea extracts were investigated. The chemical fingerprinting were carried out by high performance thin layer chromatography (HPTLC), antioxidant activity by ABTS, DPPH, FRAP radical scavenging assays, and antiogout activity by cow milk xanthine oxidase. The HPTLC fingerprint qualitatively revealed predominant amount of flavonoids. The TEAC values ranged from 45.80 to 140 µM trolox/100 g dry weight for ABTS, from 85 to 430 µM trolox/ 100 g dw DPPH, and 185 to 560 µM trolox/100 g dw for FRAP respectively. Plants used in this study was found to inhibit the toxicity, as seen from the decreased LPO and increased GSH, SOD and CAT levels. The total phenolic and flavonoid content ranged from 10.21 to 28.17 and 5.80 to 10.1 mg of gallic acid equivalents (GAE)/100 gdw respectively. The plant extracts demonstrated significant xanthine oxidase inhibitory activity at 100 g/mL and revealed an inhibition greater than 50 % and IC50 values below the standard. This effect was almost similar to the activity of allopurinol (Standard drug) against xanthine oxidase (90.2 ± 0.4 %). These plant root extract will be subjected for further extensive studies to isolate and identify their active constituents which are useful for against inflammation and gout. PMID:25237348

  14. Genetic analysis of lipolytic activities in Thermus thermophilus HB27.

    PubMed

    Leis, Benedikt; Angelov, Angel; Li, Haijuan; Liebl, Wolfgang

    2014-12-10

    The extremely thermophilic bacterium Thermus thermophilus HB27 displays lipolytic activity for the hydrolysis of triglycerides. In this study we performed a mutational in vivo analysis of esterases and lipases that confer growth on tributyrin. We interrupted 10 ORFs suspected to encode lipolytic enzymes. Two chromosomal loci were identified that resulted in reduced hydrolysis capabilities against tributyrin and various para-nitrophenyl acyl esters. By implementation of a convenient new one-step method which abstains from the use of selectable markers, a mutant strain with multiple scar-less deletions was constructed by sequentially deleting ORFs TT_C1787, TT_C0340, TT_C0341 and TT_C0904. The quadruple deletion mutant of T. thermophilus exhibited significantly lower lipolytic activity (approximately 25% residual activity compared to wild type strain) over a broad range of fatty acyl esters and had lost the ability to grow on agar plates containing tributyrin as the sole carbon source. Furthermore, we were able to determine the impact of each gene disruption on the lipolytic activity profile in this model organism and show that the esterase activity in T. thermophilus HB27 is due to a concerted action of several hydrolases having different substrate preferences and activities. The esterase-less T. thermophilus multi-deletion mutant from this study can be used as a screening and expression host for esterase genes from thermophiles or metagenomes. PMID:25102235

  15. Genetic analysis of lipolytic activities in Thermus thermophilus HB27.

    PubMed

    Leis, Benedikt; Angelov, Angel; Li, Haijuan; Liebl, Wolfgang

    2014-12-10

    The extremely thermophilic bacterium Thermus thermophilus HB27 displays lipolytic activity for the hydrolysis of triglycerides. In this study we performed a mutational in vivo analysis of esterases and lipases that confer growth on tributyrin. We interrupted 10 ORFs suspected to encode lipolytic enzymes. Two chromosomal loci were identified that resulted in reduced hydrolysis capabilities against tributyrin and various para-nitrophenyl acyl esters. By implementation of a convenient new one-step method which abstains from the use of selectable markers, a mutant strain with multiple scar-less deletions was constructed by sequentially deleting ORFs TT_C1787, TT_C0340, TT_C0341 and TT_C0904. The quadruple deletion mutant of T. thermophilus exhibited significantly lower lipolytic activity (approximately 25% residual activity compared to wild type strain) over a broad range of fatty acyl esters and had lost the ability to grow on agar plates containing tributyrin as the sole carbon source. Furthermore, we were able to determine the impact of each gene disruption on the lipolytic activity profile in this model organism and show that the esterase activity in T. thermophilus HB27 is due to a concerted action of several hydrolases having different substrate preferences and activities. The esterase-less T. thermophilus multi-deletion mutant from this study can be used as a screening and expression host for esterase genes from thermophiles or metagenomes.

  16. Transition path theory analysis of c-Src kinase activation.

    PubMed

    Meng, Yilin; Shukla, Diwakar; Pande, Vijay S; Roux, Benoît

    2016-08-16

    Nonreceptor tyrosine kinases of the Src family are large multidomain allosteric proteins that are crucial to cellular signaling pathways. In a previous study, we generated a Markov state model (MSM) to simulate the activation of c-Src catalytic domain, used as a prototypical tyrosine kinase. The long-time kinetics of transition predicted by the MSM was in agreement with experimental observations. In the present study, we apply the framework of transition path theory (TPT) to the previously constructed MSM to characterize the main features of the activation pathway. The analysis indicates that the activating transition, in which the activation loop first opens up followed by an inward rotation of the αC-helix, takes place via a dense set of intermediate microstates distributed within a fairly broad "transition tube" in a multidimensional conformational subspace connecting the two end-point conformations. Multiple microstates with negligible equilibrium probabilities carry a large transition flux associated with the activating transition, which explains why extensive conformational sampling is necessary to accurately determine the kinetics of activation. Our results suggest that the combination of MSM with TPT provides an effective framework to represent conformational transitions in complex biomolecular systems. PMID:27482115

  17. Protozoal Meningoencephalitis in Sea Otters (Enhydra lutris): a Histopathological and Immunohistochemical Study of Naturally Occuring Cases

    USGS Publications Warehouse

    Thomas, N.J.; Dubey, J.P.; Lindsay, D.S.; Cole, R.A.; Meteyer, C.U.

    2007-01-01

    Protozoal meningoencephalitis is considered to be an important cause of mortality in the California sea otter (Enhydra lutris). Thirty nine of 344 (11.3%) California (CA) and Washington state (WA) sea otters examined from 1985 to 2004 had histopathological evidence of significant protozoal meningoencephalitis. The aetiological agents and histopathological changes associated with these protozoal infections are described. The morphology of the actively multiplicative life stages of the organisms (tachyzoites for Toxoplasma gondii and merozoites for Sarcocystis neurona) and immunohistochemical labelling were used to identify infection with S. neurona (n=22, 56.4%), T. gondii (n=5, 12.8%) or dual infection with both organisms (n=12, 30.8%). Active S. neurona was present in all dual infections, while most had only the latent form of T. gondii. In S. neurona meningoencephalitis, multifocal to diffuse gliosis was widespread in grey matter and consistently present in the molecular layer of the cerebellum. In T. gondii meningoencephalitis, discrete foci of gliosis and malacia were more widely separated, sometimes incorporated pigment-laden macrophages and mineral, and were found predominantly in the cerebral cortex. Quiescent tissue cysts of T. gondii were considered to be incidental and not a cause of clinical disease and mortality. Protozoal meningoencephalitis was diagnosed more frequently in the expanding population of WA sea otters (10 of 31, 32.3%) than in the declining CA population (29 of 313, 9.3%). Among sea otters with protozoal meningoencephalitis, those that had displayed neurological signs prior to death had active S. neurona encephalitis, supporting the conclusion that S. neurona is the most significant protozoal pathogen in the central nervous system of sea otters.

  18. Multidisciplinary analysis of actively controlled large flexible spacecraft

    NASA Technical Reports Server (NTRS)

    Cooper, Paul A.; Young, John W.; Sutter, Thomas R.

    1986-01-01

    The control of Flexible Structures (COFS) program has supported the development of an analysis capability at the Langley Research Center called the Integrated Multidisciplinary Analysis Tool (IMAT) which provides an efficient data storage and transfer capability among commercial computer codes to aid in the dynamic analysis of actively controlled structures. IMAT is a system of computer programs which transfers Computer-Aided-Design (CAD) configurations, structural finite element models, material property and stress information, structural and rigid-body dynamic model information, and linear system matrices for control law formulation among various commercial applications programs through a common database. Although general in its formulation, IMAT was developed specifically to aid in the evaluation of the structures. A description of the IMAT system and results of an application of the system are given.

  19. Neutron Activation Analysis for the Demonstration of Amphibolite Rock-Weathering Activity of a Yeast

    PubMed Central

    Rades-Rohkohl, E.; Hirsch, P.; Fränzle, O.

    1979-01-01

    Neutron activation analysis was employed in a survey of weathering abilities of rock surface microorganisms. A yeast isolated from an amphibolite of a megalithic grave was found actively to concentrate, in media and in or on cells, iron and other elements when grown in the presence of ground rock. This was demonstrated by comparing a spectrum of neutron-activated amphibolite powder (particle size, 50 to 100 μm) with the spectra of neutron-activated, lyophilized yeast cells which had grown with or without amphibolite powder added to different media. The most active yeast (IFAM 1171) did not only solubilize Fe from the rock powder, but significant amounts of Co, Eu, Yb, Ca, Ba, Sc, Lu, Cr, Th, and U were also mobilized. The latter two elements occurred as natural radioactive isotopes in this amphibolite. When the yeast cells were grown with neutron-activated amphibolite, the cells contained the same elements. Furthermore, the growth medium contained Fe, Co, and Eu which had been solubilized from the amphibolite. This indicates the presence, in this yeast strain, of active rockweathering abilities as well as of uptake mechanisms for solubilized rock components. PMID:16345472

  20. Currently available useful immunohistochemical markers of renal pathology for the diagnosis of renal allograft rejection.

    PubMed

    Kanzaki, Go; Shimizu, Akira

    2015-07-01

    Renal allograft dysfunction may be induced by various causes, including alloimmune rejection, viral infection, urinary tract obstruction, calcineurin inhibitor nephrotoxicity and/or recurrent renal disease. In order to determine the underlying cause, a renal biopsy is performed and the renal transplant pathology is diagnosed using the internationally consensus Banff classification. Although a progressive understanding of allograft rejection has provided numerous immunohistochemical markers, only the C4d is regarded to be a sufficiently useful marker for antibody-mediated allograft rejection according to the Banff classification. This review summarizes currently available useful immunohistochemical markers of renal transplant pathology, including C4d, with diagnostic implications for human renal allograft rejection. In particular, we discuss immunohistochemical markers in the following three categories: immunohistochemical markers of renal pathology used to (i) analyze the mechanisms of alloimmune rejection, (ii) monitor cell injury and/or inflammation associated with rejection and (iii) identify renal components in order to improve the diagnosis of rejection. In addition, recent progress in the field of renal transplant pathology includes the development of a new method for assessing molecular pathology using OMICS analyses. As the recent findings of various studies in patients undergoing renal transplantation are very encouraging, novel immunohistochemical markers must be also developed and combined with new technologies for the diagnosis of human renal allograft rejection.

  1. Nonlinear analysis of brain activity in magnetic influenced Parkinson patients.

    PubMed

    Anninos, P A; Adamopoulos, A V; Kotini, A; Tsagas, N

    2000-01-01

    Magnetoencephalogram (MEG) recordings were obtained from the brain of patients suffering from Parkinson's disease (PD) using the Superconductive Quantum Interference Device (SQUID). For each patient the magnetic activity was recorded from a total of 64 points of the skull (32 points from each temporal lobe) as defined by a recording reference system, which is based on the 10-20 Electrode Placement System. Some of the recorded points were observed to exhibit abnormal rhythmic activity, characterized by high amplitudes and low frequencies. External magnetic stimulation (EMS) with intensity 1-7.5pT, and frequency the alpha-rhythm of the patient (8-13 Hz) was applied in the left-right temporal, frontal-occipital and vertex (2 minutes over each of the above regions) and the brain magnetic activity was recorded again. The application of the EMS resulted in rapid attenuation of the MEG activity of PD patients. Furthermore, chaotic dynamic methods were used, in order to estimate the correlation dimension D of the reconstructed phase spaces. The estimated values of D, in conjunction with the results derived from the other data analysis methods, strongly support the existence of low dimension chaotic structures in the dynamics of cortical activity of PD patients. In addition, the increased values of D of the MEG after the application of EMS when compared with the corresponding ones obtained from the MEGs prior to the EMS, suggest that the neural dynamics are strongly influenced by the application of EMS. PMID:11154103

  2. Eye movement analysis for activity recognition using electrooculography.

    PubMed

    Bulling, Andreas; Ward, Jamie A; Gellersen, Hans; Tröster, Gerhard

    2011-04-01

    In this work, we investigate eye movement analysis as a new sensing modality for activity recognition. Eye movement data were recorded using an electrooculography (EOG) system. We first describe and evaluate algorithms for detecting three eye movement characteristics from EOG signals-saccades, fixations, and blinks-and propose a method for assessing repetitive patterns of eye movements. We then devise 90 different features based on these characteristics and select a subset of them using minimum redundancy maximum relevance (mRMR) feature selection. We validate the method using an eight participant study in an office environment using an example set of five activity classes: copying a text, reading a printed paper, taking handwritten notes, watching a video, and browsing the Web. We also include periods with no specific activity (the NULL class). Using a support vector machine (SVM) classifier and person-independent (leave-one-person-out) training, we obtain an average precision of 76.1 percent and recall of 70.5 percent over all classes and participants. The work demonstrates the promise of eye-based activity recognition (EAR) and opens up discussion on the wider applicability of EAR to other activities that are difficult, or even impossible, to detect using common sensing modalities.

  3. Analysis of sprinting activities of professional soccer players.

    PubMed

    Andrzejewski, Marcin; Chmura, Jan; Pluta, Beata; Strzelczyk, Ryszard; Kasprzak, Andrzej

    2013-08-01

    The aim of the study was a detailed analysis of the sprinting activity of professional soccer players. The study involved 147 players who played in 10 matches of the 2008-09 and 2010-11 UEFA Europa League seasons. The number of performed sprints and total sprint distances covered by the players were examined using collected statistical material. Two types of sprints were distinguished based on their duration: S, short-duration sprint (below 5 seconds) and L, long-duration sprint (above 5 seconds). Additionally, sprints were classified according to their distance: 0-10, 10.1-20.0, and >20 m, respectively. The analysis of the sprinting activity of soccer players also involved their respective positions of play. The study was carried out using Amisco Pro (version. 1.0.2), one of the most comprehensive up-to-date computer systems for match analysis. The statistical analysis revealed that the mean total sprint distance covered by players (≥24 km·h) amounted to 237 ± 123 m. With regard to the position of play, the forwards covered the longest sprint distance (345 ± 129 m), that is, 9% longer than midfielders (313 ± 119 m) and over 100% longer than central midfielders (167 ± 87 m). The average number of sprints performed by the soccer players was 11.2 ± 5.3. It should also be emphasized that about 90% of sprints performed by professional soccer players were shorter than 5 seconds, whereas only 10% were longer than 5 seconds. Analysis of physical loads of soccer players during matches can be useful for individualization of training of soccer players' speed capabilities. It is an essential instrument of modern planning and application of training loads.

  4. Mineral exploration and soil analysis using in situ neutron activation

    USGS Publications Warehouse

    Senftle, F.E.; Hoyte, A.F.

    1966-01-01

    A feasibility study has been made to operate by remote control an unshielded portable positive-ion accelerator type neutron source to induce activities in the ground or rock by "in situ" neutron irradiation. Selective activation techniques make it possible to detect some thirty or more elements by irradiating the ground for periods of a few minutes with either 3-MeV or 14-MeV neutrons. The depth of penetration of neutrons, the effect of water content of the soil on neutron moderation, gamma ray attenuation in the soil and other problems are considered. The analysis shows that, when exploring for most elements of economic interest, the reaction 2H(d,n)3He yielding ??? 3-MeV neutrons is most practical to produce a relatively uniform flux of neutrons of less than 1 keV to a depth of 19???-20???. Irradiation with high energy neutrons (??? 14 MeV) can also be used and may be better suited for certain problems. However, due to higher background and lower sensitivity for the heavy minerals, it is not a recommended neutron source for general exploration use. Preliminary experiments have been made which indicate that neutron activation in situ is feasible for a mineral exploration or qualititative soil analysis. ?? 1976.

  5. Geometric Nonlinear Finite Element Analysis of Active Fibre Composite Bimorphs

    NASA Astrophysics Data System (ADS)

    Kernaghan, Robert

    Active fibre composite-actuated bimorphic actuators were studied in order to measure deflection performance. The deflection of the actuators was a function of the actuating electric potential applied to the active material as well as the magnitude of the axial preload applied to the bimorphic structure. This problem required the use of geometric nonlinear modeling techniques. Geometric nonlinear finite element analysis was undertaken to determine the deflection performance of Macro Fibre Composite (MFC)- and Hollow Active Fibre (HAFC)-actuated bimorphic structures. A physical prototype MFC-actuated bimorphic structure was manufactured in order to verify the results obtained by the finite element analysis. Theses analyses determined that the bimorphic actuators were capable of significant deflection. The analyses determined that the axial preload of the bimorphic actuators significantly amplified the deflection performance of the bimorphic actuators. The deflection performance of the bimorphic actuators suggest that they could be candidates to act as actuators for the morphing wing of a micro unmanned air vehicle.

  6. Detection of MYB Alterations and Other Immunohistochemical Markers in Primary Cutaneous Adenoid Cystic Carcinoma.

    PubMed

    North, Jeffrey P; McCalmont, Timothy H; Fehr, André; van Zante, Annemieke; Stenman, Göran; LeBoit, Philip E

    2015-10-01

    Adenoid cystic carcinoma (ACC) can arise in several organs, and prognosis is highly dependent on the primary tumor site. Primary cutaneous ACC has an excellent prognosis compared with salivary or lacrimal ACC. Activation of MYB by gene fusion or other mechanisms has been found in salivary, breast, and lacrimal ACCs but has not been described in cutaneous ACC. We analyzed the histopathologic and immunohistochemical features of 19 primary cutaneous ACCs, 2 periorbital ACCs, and 12 salivary gland ACCs and assessed for MYB activation in primary cutaneous ACC by immunohistochemistry and molecular methods. The presence of perineural invasion differed significantly among ACCs of various sites (83% salivary, 50% eyelid, 11% skin, P=0.0002). Over 90% of all ACCs were grade 1 or 2 and exhibited diffuse (>50%) positivity with CD117, SOX-10, and smooth muscle actin immunostains. CK15 and vimentin showed diffuse positivity in 36% and 57% of cutaneous ACCs, respectively, and were negative or only focally positive in all salivary ACCs (P=0.04 and 0.002). Six of the 11 cutaneous and periorbital ACCs tested with reverse transcriptase polymerase chain reaction and/or fluorescence in situ hybridization had MYB rearrangements including 2 cases that expressed MYB-NFIB fusion transcripts. Diffuse expression of MYB protein assessed by immunostaining was present in 8 of 9 cutaneous ACCs, including cases both with and without MYB rearrangements. These results indicate that cutaneous ACCs possess the same types of MYB alterations as ACCs of other anatomic sites. Vimentin and CK15 appear to have some discriminatory value in differentiating between primary cutaneous and salivary gland ACCs. PMID:26076064

  7. Immunohistochemical localisation and molecular expression of the steroidogenic enzyme cytochrome P450 17α-hydroxylase /C(17,20)-lyase in the vestibular nuclei of adult male rats.

    PubMed

    Manca, P; Mulliri, G; Burrai, G P; Pirino, S; Mameli, O

    2011-05-01

    Many biologically active neurosteroids, including dehydroepiandrosterone (DHEA), are synthesised in the brain. DHEA is a potent endogenous modulator of several neuronal functions, and alterations of DHEA are correlated with various neurobiological deficits. The cytochrome P450 17α-hydroxylase/C(17,20)-lyase (P450C(17) ) plays a pivotal role in the synthesis of DHEA from pregnenolone and progesterone. We investigated the immunohistochemical localisation and molecular expression of P450C(17) in the superior, lateral, medial and inferior vestibular nuclei (VCN) of adult male rats by western blotting and indirect immunofluorescence analysis. Immunoreactive P450C(17) was widely distributed in all VCN and the expression of P450C(17) was confirmed by western blot analysis. The present study demonstrates, for the first time, the presence and anatomical distribution of P450C(17) in the VCN. Given that neurosteroids can modulate neuronal activities in the medial vestibular nucleus, DHEA synthesised in the VCN may play an important role in the control of specific activities at this level.

  8. Immunohistochemical evaluation of myofibroblasts in odontogenic cysts and tumors: A comparative study

    PubMed Central

    Syamala, Deepa; Suresh, Rakesh; Janardhanan, Mahija; Savithri, Vindhya; Anand, Prem P; Jose, Amrutha

    2016-01-01

    Context: Myofibroblasts are fibroblasts with smooth muscle-like features characterized by the presence of a contractile apparatus and found in the connective tissue stroma of normal tissues such as blood vessels and lymph nodes. They are now thought to play a role in the synthesis and reorganization of extracellular matrix, which could contribute to the aggressive biologic behavior of the lesions. Aims: To compare the mean number of stromal myofibroblasts in dentigerous cysts (DCs), keratocystic odontogenic tumor (KCOT) and ameloblastoma; and to derive a correlation between the stromal myofibroblasts and the known biologic behavior of the lesions. Settings and Design: A cross-sectional immunohistochemical analysis of cases of DC, KCOT and ameloblastoma. Materials and Methods: Twenty paraffin-embedded tissue blocks each of DC, KCOT and multicystic ameloblastoma were selected for the study and diagnosis confirmed through hematoxylin and eosin staining. Tissue sections were analyzed for the number of myofibroblasts using alpha smooth muscle actin (α-SMA) immunostaining. Statistical Analysis: Differences in the mean number of α-SMA positive cells in each group were analyzed using one-way ANOVA test. Intergroup comparisons of mean values of α-SMA positive cells were performed using Mann-Whitney U-test. Results: Ameloblastoma showed the highest number of myofibroblasts, whereas DC showed the lowest. Among the groups, there were significant differences between the myofibroblast counts among DC and KCOT and between DC and ameloblastoma, whereas the difference in counts was not statistically significant between KCOT and ameloblastoma. A positive correlation was observed between the myofibroblast count and the known biologic behavior of the lesions. Conclusion: Myofibroblasts may act in close association with the epithelial cells to bring about changes in stromal microenvironment, favorable to the growth and progression of the lesion. They may be of great value in

  9. Quantitative evaluation of microvessel density using CD34 in clinical variants of ameloblastoma: An immunohistochemical study

    PubMed Central

    Pereira, Treville; Dodal, Shashibhushan; Tamgadge, Avinash; Bhalerao, Sudhir; Tamgadge, Sandhya

    2016-01-01

    Background: Odontogenic epithelium plays an important role in the histogenesis of odontogenic tumors of the jaws. Ameloblastomas, which arise from odontogenic epithelium, are considered benign with little tendency to metastasize. Tumors require an adequate supply of oxygen and a way to remove their waste products. This can be achieved by angiogenesis. In situ quantification of the microvessel density (MVD) is a usual method for assessing angiogenesis. Moreover, angiogenesis may differ in subtypes of ameloblastomas and could play a role in determining the pattern of tumor growth. Aim: The aim of the present study was to demonstrate the expression of cluster of differentiation (CD34) in variants of ameloblastomas and to correlate and compare their expression to the aggressive behavior. Materials and Methods: A retrospective cross-sectional study which included forty paraffin blocks was conducted after obtaining ethical committee clearance. Ten cases of pyogenic granuloma were used as a positive control and thirty cases were of solid multicystic ameloblastoma (SMA), unicystic ameloblastoma (UA) and desmoplastic ameloblastomas. Angiogenesis was assessed using CD34 antigen and was immunohistochemically localized. Statistical analysis was carried out for comparative analysis with the help of ANOVA test, Kolmogorov–Smirnov test and least significance difference test. Results: A significant correlation was obtained between the MVD of all the three variants, i.e., SMA, UA and desmoplastic ameloblastomas which was statistically significant (P < 0.05). Conclusion: Increased MVD in the three variants, i.e., SMA, UA and desmoplastic ameloblastoma seen in the present study could suggest that the angiogenesis has an important role in tumor progression and aggressiveness of ameloblastomas. PMID:27194862

  10. Immunohistochemical evaluation of myofibroblasts in odontogenic cysts and tumors: A comparative study

    PubMed Central

    Syamala, Deepa; Suresh, Rakesh; Janardhanan, Mahija; Savithri, Vindhya; Anand, Prem P; Jose, Amrutha

    2016-01-01

    Context: Myofibroblasts are fibroblasts with smooth muscle-like features characterized by the presence of a contractile apparatus and found in the connective tissue stroma of normal tissues such as blood vessels and lymph nodes. They are now thought to play a role in the synthesis and reorganization of extracellular matrix, which could contribute to the aggressive biologic behavior of the lesions. Aims: To compare the mean number of stromal myofibroblasts in dentigerous cysts (DCs), keratocystic odontogenic tumor (KCOT) and ameloblastoma; and to derive a correlation between the stromal myofibroblasts and the known biologic behavior of the lesions. Settings and Design: A cross-sectional immunohistochemical analysis of cases of DC, KCOT and ameloblastoma. Materials and Methods: Twenty paraffin-embedded tissue blocks each of DC, KCOT and multicystic ameloblastoma were selected for the study and diagnosis confirmed through hematoxylin and eosin staining. Tissue sections were analyzed for the number of myofibroblasts using alpha smooth muscle actin (α-SMA) immunostaining. Statistical Analysis: Differences in the mean number of α-SMA positive cells in each group were analyzed using one-way ANOVA test. Intergroup comparisons of mean values of α-SMA positive cells were performed using Mann-Whitney U-test. Results: Ameloblastoma showed the highest number of myofibroblasts, whereas DC showed the lowest. Among the groups, there were significant differences between the myofibroblast counts among DC and KCOT and between DC and ameloblastoma, whereas the difference in counts was not statistically significant between KCOT and ameloblastoma. A positive correlation was observed between the myofibroblast count and the known biologic behavior of the lesions. Conclusion: Myofibroblasts may act in close association with the epithelial cells to bring about changes in stromal microenvironment, favorable to the growth and progression of the lesion. They may be of great value in

  11. Immunohistochemical (Ki-67) study of endometrial maturation in mice after use of phosphodiesterase type 5 inhibitor

    PubMed Central

    Rashidi, Bahman; Rad, Jafar Soleimani; Rad, Leila Roshangar

    2015-01-01

    Background: Uterine receptivity for the implantation is a complicated process, that ovarian factors (hormonal), endometrium and embryo simultaneously are involved in this phenomenon. A successful implantation needs appropriate development of the endometrium. Furthermore, embryo must be capable of reacting with the endometrium and producing suitable adhesion molecules. This study aimed to examine one of endometrial maturation indices in mice before implantation, i.e., proliferation of stromal cells. Materials and Methods: A total of 40 adult female mice were divided into four groups: Control, gonadotropin, gonadotropin + progesterone, and gonadotropin + sildenafil citrate. The three experimental groups were first injected 7.5 IU of human chorionic gonadotropin (HCG) and then 7.5 IU of human menopausal gonadotropin (HMG). Then, every two female mice were placed in a cage with a male mouse for mating. Two groups were injected 1 mg of progesterone and 3 mg/kg of sildenafil citrate at intervals of 24, 48, and 72 h after injection of HMG. After 96 h, all the mice were killed, and their uterine samples subjected to tissue passage and prepared for analysis. Immunohistochemical method, Ki-67, and stromal mitotic cell count were used in this study. Results: Our observations in all groups showed changes in the luminal epithelium. ANOVA analysis Ki-67-positive stromal cells among all groups were not statistically significant. Conclusion: The results showed that administration of HMG and HCG following that of progesterone and sildenafil citrate could change the indices of endometrial maturation, and they were not involved in the phase immediately before implantation in stromal mitotic index. PMID:26380239

  12. Immunohistochemical Comparison of IL-36 and the IL-23/Th17 Axis of Generalized Pustular Psoriasis and Acute Generalized Exanthematous Pustulosis

    PubMed Central

    Song, Hyo Sang; Kim, Sang Jin; Park, Tae-In; Jang, Yong Hyun

    2016-01-01

    Background Cutaneous pustular disorders include generalized pustular psoriasis (GPP) and acute generalized exanthematous pustulosis (AGEP). Objective To identify differences between GPP and AGEP, here we immunohistochemically evaluated interleukin (IL)-36 and the IL-23/Th17 axis. Methods This retrospective comparative immunohistochemical study was completed using 11 biopsies of 11 cases of GPP and 11 biopsies of 11 cases of AGEP. Through staining with the anti-IL-36-alpha (IL-36α), anti-IL-36 receptor antagonist (IL-36Ra), anti-nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), anti-IL-23, anti-IL-17, and anti-IL-8 antibodies, main expression location and intensity were visualized in the epidermis and dermis. Results In both diseases, diffuse IL-36α expression was observed in the epidermis. IL-36Ra expression was observed in the dermal perivascular area as well as in the epidermis. NF-κB expression was observed in the epidermis and perivascular dermal area. Diffuse IL-23 and IL-17 expression was seen in the whole epidermis and the perivascular dermal area. IL-8 was expressed in the subcorneal pustules and parakeratotic area. Contrary to other cytokines, IL-23 expression in the epidermis of patients with GPP was more intense than only that in patients with AGEP. Conclusion Common pathomechanisms might exist in the development of GPP and AGEP based on these immunohistochemical results, but further studies are needed. PMID:27489427

  13. Analysis of antifreeze protein activity using colorimetric gold nanosensors

    NASA Astrophysics Data System (ADS)

    Jing, Xu; Choi, Ho-seok; Park, Ji-In; Kim, Young-Pil

    2015-07-01

    High activity and long stability of antifreeze proteins (AFPs), also known as ice-binding proteins (IBPs), are necessary for exerting their physiological functions in biotechnology and cryomedicine. Here we report a simple analysis of antifreeze protein activity and stability based on self-assembly of gold nanoparticles (AuNPs) via freezing and thawing cycles. While the mercaptosuccinic acid-capped AuNP (MSA-AuNP) was easily self-assembled after a freezing/thawing cycle, due to the mechanical attack of ice crystal on the MSA-AuNP surface, the presence of AFP impeded the self-assembly of MSA-AuNP via the interaction of AFP with ice crystals via freezing and thawing cycles, which led to a strong color in the MSA-AuNP solution. As a result, the aggregation parameter (E520/E650) of MSA-AuNP showed the rapid detection of both activity and stability of AFPs. We suggest that our newly developed method is very suitable for measuring antifreeze activity and stability in a simple and rapid manner with reliable quantification.

  14. [Kinetic analysis of additive effect on desulfurization activity].

    PubMed

    Han, Kui-hua; Zhao, Jian-li; Lu, Chun-mei; Wang, Yong-zheng; Zhao, Gai-ju; Cheng, Shi-qing

    2006-02-01

    The additive effects of A12O3, Fe2O3 and MnCO3 on CaO sulfation kinetics were investigated by thermogravimetic analysis method and modified grain model. The activation energy (Ea) and the pre-exponential factor (k0) of surface reaction, the activation energy (Ep) and the pre-exponential factor (D0) of product layer diffusion reaction were calculated according to the model. Additions of MnCO3 can enhance the initial reaction rate, product layer diffusion and the final CaO conversion of sorbents, the effect mechanism of which is similar to that of Fe2O3. The method based isokinetic temperature Ts and activation energy can not estimate the contribution of additive to the sulfation reactivity, the rate constant of the surface reaction (k), and the effective diffusivity of reactant in the product layer (Ds) under certain experimental conditions can reflect the effect of additives on the activation. Unstoichiometric metal oxide may catalyze the surface reaction and promote the diffusivity of reactant in the product layer by the crystal defect and distinct diffusion of cation and anion. According to the mechanism and effect of additive on the sulfation, the effective temperature and the stoichiometric relation of reaction, it is possible to improve the utilization of sorbent by compounding more additives to the calcium-based sorbent.

  15. CMS dashboard for monitoring of the user analysis activities

    NASA Astrophysics Data System (ADS)

    Karavakis, Edward; Andreeva, Julia; Maier, Gerhild; Khan, Akram

    2012-12-01

    The CMS Virtual Organisation (VO) uses various fully distributed job submission methods and execution backends. The CMS jobs are processed on several middleware platforms such as the gLite, the ARC and the OSG. Up to 200,000 CMS jobs are submitted daily to the Worldwide LHC Computing Grid (WLCG) infrastructure and this number is steadily growing. These mentioned factors increase the complexity of the monitoring of the user analysis activities within the CMS VO. Reliable monitoring is an aspect of particular importance; it is a vital factor for the overall improvement of the quality of the CMS VO infrastructure.

  16. Fractal analysis of granular activated carbons using isotherm data

    SciTech Connect

    Khalili, N.R.; Pan, M.; Sandi, G.

    1997-08-01

    Utilization of adsorption on solid surfaces was exercised for the first time in 1785. Practical application of unactivated carbon filters, and powdered carbon were first demonstrated in the American water treatment plant, and a municipal treatment plant in New Jersey, in 1883 and 1930, respectively. The use of activated carbon became widespread in the next few decades. At present, adsorption on carbons has a wide spread application in water treatment and removal of taste, odor, removal of synthetic organic chemicals, color-forming organics, and desinfection by-products and their naturally occurring precursors. This paper presents an analysis of the surface fractal dimension and adsorption capacity of a group of carbons.

  17. Activation analysis of admixtures in certain semiconductive materials

    NASA Technical Reports Server (NTRS)

    Artyukhin, P. I.; Gilbert, E. P.; Pronin, V. A.

    1978-01-01

    The use of extractions and chromatographic operations to separate macrobases, and to divide elements into groups convenient for gamma-spectrometric analysis is discussed. Methods are described for the activation detection of some impurities in silicon, arsenic, thallium, and trichloromethylsilane, on the basis of the extraction properties of bis(2-chlorethyl ether) and dimethylbenzylalkylammonium chloride. A schematic diagram of the extraction separation of elements-admixture is presented showing the aqueous and organic phases. The content percentage of the various elements are given in tables.

  18. Toxicogenomic analysis of pharmacological active coumarins isolated from Calophyllum brasiliense

    PubMed Central

    Gomez-Verjan, J.C.; Estrella-Parra, E.A.; Gonzalez-Sanchez, I.; Rivero-Segura, N.A.; Vazquez-Martinez, R.; Magos-Guerrero, G.; Mendoza-Villanueva, D.; Cerbón-Cervantes, M.A.; Reyes-Chilpa, R.

    2015-01-01

    Calophyllum brasiliense (Calophyllaceae) is a tropical rain forest tree, mainly distributed in South and Central America. It is an important source of bioactive natural products like, for instance soulatrolide, and mammea type coumarins. Soulatrolide is a tetracyclic dipyranocoumarins and a potent inhibitor of HIV-1 reverse transcriptase and Mycobacterium tuberculosis. Mammea A/BA and A/BB coumarins, pure or as a mixture, are highly active against several leukemia cell lines, Trypanosoma cruzi and Leishmania amazonensis. In the present work, a toxicogenomic analysis of Soulatrolide and Mammea A/BA + A/BB (3:1) mixture was performed in order to validate the toxicological potential of this type of compounds. Soulatrolide or mixture of mammea A/BA + A/BB (3:1) was administered orally to male mice (CD-1) at dose of 100 mg/kg/daily, for 1 week. After this time, mice were sacrificed, and RNA extracted from the liver of treated animals. Transcriptomic analysis was performed using Affymetrix Mouse Gene 1.0 ST Array. Robust microarray analysis (RMA) and two way ANOVA test revealed for mammea mixture treatment 46 genes upregulated and 72 downregulated genes; meanwhile, for soulatrolide 665 were upregulated and 1077 downregulated genes. Enrichment analysis for such genes revealed that in both type of treatments genetic expression were mainly involved in drug metabolism. Overall results indicate a safety profile. The microarray data complies with MIAME guidelines and are deposited in GEO under accession number GSE72755. PMID:26697389

  19. Toxicogenomic analysis of pharmacological active coumarins isolated from Calophyllum brasiliense.

    PubMed

    Gomez-Verjan, J C; Estrella-Parra, E A; Gonzalez-Sanchez, I; Rivero-Segura, N A; Vazquez-Martinez, R; Magos-Guerrero, G; Mendoza-Villanueva, D; Cerbón-Cervantes, M A; Reyes-Chilpa, R

    2015-12-01

    Calophyllum brasiliense (Calophyllaceae) is a tropical rain forest tree, mainly distributed in South and Central America. It is an important source of bioactive natural products like, for instance soulatrolide, and mammea type coumarins. Soulatrolide is a tetracyclic dipyranocoumarins and a potent inhibitor of HIV-1 reverse transcriptase and Mycobacterium tuberculosis. Mammea A/BA and A/BB coumarins, pure or as a mixture, are highly active against several leukemia cell lines, Trypanosoma cruzi and Leishmania amazonensis. In the present work, a toxicogenomic analysis of Soulatrolide and Mammea A/BA + A/BB (3:1) mixture was performed in order to validate the toxicological potential of this type of compounds. Soulatrolide or mixture of mammea A/BA + A/BB (3:1) was administered orally to male mice (CD-1) at dose of 100 mg/kg/daily, for 1 week. After this time, mice were sacrificed, and RNA extracted from the liver of treated animals. Transcriptomic analysis was performed using Affymetrix Mouse Gene 1.0 ST Array. Robust microarray analysis (RMA) and two way ANOVA test revealed for mammea mixture treatment 46 genes upregulated and 72 downregulated genes; meanwhile, for soulatrolide 665 were upregulated and 1077 downregulated genes. Enrichment analysis for such genes revealed that in both type of treatments genetic expression were mainly involved in drug metabolism. Overall results indicate a safety profile. The microarray data complies with MIAME guidelines and are deposited in GEO under accession number GSE72755.

  20. Detection of estrogen receptors in the trigonum and urinary bladder with an immunohistochemical technique.

    PubMed

    Bussolati, G; Tizzani, A; Casetta, G; Cassoni, P; Pacchioni, D; Piana, P; Revelli, A; Massobrio, M

    1990-09-01

    In order to detect estrogen receptors (ER) in the female bladder, 5 premenopausal and 10 postmenopausal women affected by gynecological diseases were submitted to cystoscopy, during which both the trigonum and the bladder lateral wall were biopsied. A new, simple, cost-effective immunohistochemical technique was employed to stain the estrogen specific binding sites. ER were found in the trigonum of 3 premenopausal and 4 postmenopausal subjects, while the bladder lateral wall was always ER-negative. A comparison with previously used ER detection methods and a discussion of further hypothetical applications of the immunohistochemical technique in the study of the lower female urogenital tract are reported.

  1. Variability in Immunohistochemical Detection of Programmed Death Ligand 1 (PD-L1) in Cancer Tissue Types

    PubMed Central

    Scognamiglio, Giosuè; De Chiara, Anna; Di Bonito, Maurizio; Tatangelo, Fabiana; Losito, Nunzia Simona; Anniciello, Annamaria; De Cecio, Rossella; D’Alterio, Crescenzo; Scala, Stefania; Cantile, Monica; Botti, Gerardo

    2016-01-01

    In normal cell physiology, programmed death 1 (PD-1) and its ligand, PD-L1, play an immunoregulatory role in T-cell activation, tolerance, and immune-mediated tissue damage. The PD-1/PD-L1 pathway also plays a critical role in immune escape of tumor cells and has been demonstrated to correlate with a poor prognosis of patients with several types of cancer. However, recent reports have revealed that the immunohistochemical (IHC) expression of the PD-L1 in tumor cells is not uniform for the use of different antibodies clones, with variable specificity, often doubtful topographical localization, and with a score not uniquely defined. The purpose of this study was to analyze the IHC expression of PD-L1 on a large series of several human tumors to correctly define its staining in different tumor tissues. PMID:27213372

  2. Physical Workload Analysis Among Small Industry Activities Using Postural Data.

    PubMed

    Ahasan; Väyrynen; Kirvesoja

    1996-01-01

    Small industry workers are often involved in manual handling operations that require awkward body postures, so musculoskeletal disorders and occupational injuries are a major problem. In this study, various types of tasks were recorded with a video camera to chart and analyze different postures by computerized OWAS (Ovako Working Posture Analysing System). Collected data showed that poor postures were adopted, not only for lifting or hammering operation, but also for other tasks; mostly with bent and twisted back. The main aim was to determine the physical workload by identifying harmful postures and to develop recommendations for improving the existing situation. Forty-eight male workers from 8 different units (mean age: 37) participated. The performed activities were then divided into 26 sub-tasks. Altogether 1534 postures were selected for analysis. Then they were classified into different OAC (OWAS Action Categories). From all the observation, unhealthy postures, for which corrective measures had to be considered immediately (i.e., 10.6% classified as OAC III and 3.3%--as OAC IV) were found. The applied method was useful in determining the physical workload by locating potential activities due to harmful postures, providing a detailed description with analysis, and suggesting successful means to reduce postural load.

  3. Extravehicular Activity System Sizing Analysis Tool (EVAS_SAT)

    NASA Technical Reports Server (NTRS)

    Brown, Cheryl B.; Conger, Bruce C.; Miranda, Bruno M.; Bue, Grant C.; Rouen, Michael N.

    2007-01-01

    An effort was initiated by NASA/JSC in 2001 to develop an Extravehicular Activity System Sizing Analysis Tool (EVAS_SAT) for the sizing of Extravehicular Activity System (EVAS) architecture and studies. Its intent was to support space suit development efforts and to aid in conceptual designs for future human exploration missions. Its basis was the Life Support Options Performance Program (LSOPP), a spacesuit and portable life support system (PLSS) sizing program developed for NASA/JSC circa 1990. EVAS_SAT estimates the mass, power, and volume characteristics for user-defined EVAS architectures, including Suit Systems, Airlock Systems, Tools and Translation Aids, and Vehicle Support equipment. The tool has undergone annual changes and has been updated as new data have become available. Certain sizing algorithms have been developed based on industry standards, while others are based on the LSOPP sizing routines. The sizing algorithms used by EVAS_SAT are preliminary. Because EVAS_SAT was designed for use by members of the EVA community, subsystem familiarity on the part of the intended user group and in the analysis of results is assumed. The current EVAS_SAT is operated within Microsoft Excel 2003 using a Visual Basic interface system.

  4. Modeling place field activity with hierarchical slow feature analysis

    PubMed Central

    Schönfeld, Fabian; Wiskott, Laurenz

    2015-01-01

    What are the computational laws of hippocampal activity? In this paper we argue for the slowness principle as a fundamental processing paradigm behind hippocampal place cell firing. We present six different studies from the experimental literature, performed with real-life rats, that we replicated in computer simulations. Each of the chosen studies allows rodents to develop stable place fields and then examines a distinct property of the established spatial encoding: adaptation to cue relocation and removal; directional dependent firing in the linear track and open field; and morphing and scaling the environment itself. Simulations are based on a hierarchical Slow Feature Analysis (SFA) network topped by a principal component analysis (ICA) output layer. The slowness principle is shown to account for the main findings of the presented experimental studies. The SFA network generates its responses using raw visual input only, which adds to its biological plausibility but requires experiments performed in light conditions. Future iterations of the model will thus have to incorporate additional information, such as path integration and grid cell activity, in order to be able to also replicate studies that take place during darkness. PMID:26052279

  5. Rational Manual and Automated Scoring Thresholds for the Immunohistochemical Detection of TP53 Missense Mutations in Human Breast Carcinomas.

    PubMed

    Taylor, Nicholas J; Nikolaishvili-Feinberg, Nana; Midkiff, Bentley R; Conway, Kathleen; Millikan, Robert C; Geradts, Joseph

    2016-07-01

    Missense mutations in TP53 are common in human breast cancer, have been associated with worse prognosis, and may predict therapy effect. TP53 missense mutations are associated with aberrant accumulation of p53 protein in tumor cell nuclei. Previous studies have used relatively arbitrary cutoffs to characterize breast tumors as positive for p53 staining by immunohistochemical assays. This study aimed to objectively determine optimal thresholds for p53 positivity by manual and automated scoring methods using whole tissue sections from the Carolina Breast Cancer Study. p53-immunostained slides were available for 564 breast tumors previously assayed for TP53 mutations. Average nuclear p53 staining intensity was manually scored as negative, borderline, weak, moderate, or strong and percentage of positive tumor cells was estimated. Automated p53 signal intensity was measured using the Aperio nuclear v9 algorithm combined with the Genie histology pattern recognition tool and tuned to achieve optimal nuclear segmentation. Receiver operating characteristic curve analysis was performed to determine optimal cutoffs for average staining intensity and percent cells positive to distinguish between tumors with and without a missense mutation. Receiver operating characteristic curve analysis demonstrated a threshold of moderate average nuclear staining intensity as a good surrogate for TP53 missense mutations in both manual (area under the curve=0.87) and automated (area under the curve=0.84) scoring systems. Both manual and automated immunohistochemical scoring methods predicted missense mutations in breast carcinomas with high accuracy. Validation of the automated intensity scoring threshold suggests a role for such algorithms in detecting TP53 missense mutations in high throughput studies.

  6. Behavioural Activation for Depression; An Update of Meta-Analysis of Effectiveness and Sub Group Analysis

    PubMed Central

    Ekers, David; Webster, Lisa; Van Straten, Annemieke; Cuijpers, Pim; Richards, David; Gilbody, Simon

    2014-01-01

    Background Depression is a common, disabling condition for which psychological treatments are recommended. Behavioural activation has attracted increased interest in recent years. It has been over 5 years since our meta-analyses summarised the evidence supporting and this systematic review updates those findings and examines moderators of treatment effect. Method Randomised trials of behavioural activation for depression versus controls or anti-depressant medication were identified using electronic database searches, previous reviews and reference lists. Data on symptom level and study level moderators were extracted and analysed using meta-analysis, sub-group analysis and meta-regression respectively. Results Twenty six randomised controlled trials including 1524 subjects were included in this meta-analysis. A random effects meta-analysis of symptom level post treatment showed behavioural activation to be superior to controls (SMD −0.74 CI −0.91 to −0.56, k = 25, N = 1088) and medication (SMD −0.42 CI −0.83 to-0.00, k = 4, N = 283). Study quality was low in the majority of studies and follow- up time periods short. There was no indication of publication bias and subgroup analysis showed limited association between moderators and effect size. Conclusions The results in this meta-analysis support and strengthen the evidence base indicating Behavioural Activation is an effective treatment for depression. Further high quality research with longer term follow-up is needed to strengthen the evidence base. PMID:24936656

  7. Ultrastructural immunohistochemical localization of virus in acute and chronic demyelinating Theiler's virus infection.

    PubMed Central

    Dal Canto, M. C.; Lipton, H. L.

    1982-01-01

    Mice experimentally infected with Theiler's murine encephalomyelitis virus (TMEV) develop a persistent infection of the central nervous system (CNS). The most striking feature of this infection is the occurrence of inflammatory primary demyelination in the spinal cord white matter. The pathogenesis of myelin degeneration in this model has not been clarified, but morphologic and immunologic data suggest that the host immune response plays a major role in the production of myelin injury. Because of low virus titers in infected adult mice and of the small size of TMEV, virus particles have never been observed in this demyelinating model. Yet elucidation of the types of cells in the CNS supporting virus replication would be important for a better understanding of both virus persistence and virus-induced demyelinating pathology. The present paper is a sequential study of the localization of TMEV in the spinal cord in infected mice by ultrastructural immunohistochemical techniques. Results indicate that virus replication is mainly in neurons during the acute phase of the disease, while in the chronic phase viral inclusions are mainly found in macrophages in and around demyelinating lesions. Other cells are also infected, but to a lesser degree. In the neuronal system both axoplasmic and dendritic flow appear to facilitate the spread of virus in the CNS. In macrophages, the presence of virus particles and the association of virus with altered components of the cytoskeleton support active virus production rather than simple internalization. The macrophage appears to play an important role in both the establishment of virus persistence and in the process of demyelination in this animal model. Images Figure 1 and 2 Figure 3 and 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 Figure 10-12 Figure 13 Figure 14 Figure 15 and 16 PMID:6275708

  8. Betacellulin in Chronic Periodontitis Patients With and Without Type 2 Diabetes Mellitus: An Immunohistochemical Study

    PubMed Central

    Kalburgi, Nagaraj B.; Bilichodmath, Shivaprasad; Warad, Shivaraj B.; Ugale, Mahesh S.; Koregol, Arati C.; Bijjargi, Shobha C.

    2015-01-01

    Background The host immune response to bacterial dental plaque determines periodontal disease susceptibility by increasing the secretion of inflammatory cytokines. The Epidermal Growth Factor family cytokines stimulate proliferation and keratinization of cells in dermis and oral epithelium. Epidermal Growth Factor family consists of Amphiregulin, Betacellulin, Epiregulin, Epigen, Heparin binding Epidermal Growth Factor like growth factor and transforming Growth Factor-alpha. Aim The current study aimed to investigate expression of Betacellulin in chronic periodontitis patients with and without type 2 diabetes mellitus and thereby assessing role of betacellulin in periodontal health and disease. Materials and Methods Present study comprised of 90 participants, age ranges from 18 to 60-year-old, for the period of March 2010 to May 2011. Participants were categorized into three groups based Gingival index (GI), probing depth (PD) and clinical attachment loss (CA Loss). Group 1 consisted 30 individuals with clinically healthy periodontium, Group-2 consisted 30 individuals with GI>1, PD≥5 mm, and CA Loss>3 mm. Group-3 (Chronic Periodontitis with type 2 diabetes mellitus) consisted 30 with GI >1, PD≥5 mm, and CA Loss>3 mm. Immunohistochemical localization and quantification of Betacellulin was done in gingival tissue samples from all groups. Results Data showed expression of Betacellulin were higher in chronic periodontitis as compared to healthy. A positive correlation found in Betacellulin expression and Probing Depth in chronic periodontitis. Conclusion This footmark study impacts the role of Betacellulin in pathogenesis and progression of periodontal disease which will help in exploration of novel immunotherapeutic strategies and immunological research activity in this field. PMID:26673371

  9. Comparison of immunohistochemical expression of CD10 in keratocystic odontogenic tumor and ameloblastoma

    PubMed Central

    Hormozi, Elham; Fard, Vahid Nourollahi; Naseri, Mohammad Ali; Jahromi, Nima Haghighat; Keshani, Forooz

    2016-01-01

    Background: Odontogenic keratocyst (OKC), also called keratocystic odontogenic tumor (KCOT), is a developmental lesion which should be carefully monitored and it exhibits development mechanisms and biologic behaviors different from those of other more common lesions such as dentigerous and radicular cysts. CD10 antigen is a cell surface metalloendopeptidase, which inactivates various peptides that are physiologically active. Studies have shown that increase in the expression of CD10 in the stromal cells helps the progression of the tumor. Ameloblastoma (AB) is a local invasive tumor and given the role of supporting connective tissue stroma in the aggression and progression. The aim of the present study was to comparatively evaluate the expression of CD10 in the connective tissue stroma of AB and OKC as a KCOT. Materials and Methods: In this retrospective, cross-sectional study, 14 paraffin blocks of KCOT and 9 of AB (7 multicystic and 2 unicystic) were evaluated with CD10 immunohistochemical expression in the connective tissue stroma of AB and the connective tissue wall of KCOT. The data were analyzed with Fisher's exact test (P < 0.05). Results: In 8 samples of 9 AB and in 13 samples of 14 KOT lesions, expression of CD10 was shown. Fisher's exact test did not reveal any significant differences between these two lesions in the expression of CD10 (P = 0.64). Conclusion: The results of this study propose that high expression rate of CD10 might be one of the reasons for the aggressive behavior of AB and high recurrence rate of OKC and reinforce the classification of OKC as an odontogenic tumor. PMID:27076824

  10. Ultrastructural immunohistochemical localization of virus in acute and chronic demyelinating Theiler's virus infection.

    PubMed

    Dal Canto, M C; Lipton, H L

    1982-01-01

    Mice experimentally infected with Theiler's murine encephalomyelitis virus (TMEV) develop a persistent infection of the central nervous system (CNS). The most striking feature of this infection is the occurrence of inflammatory primary demyelination in the spinal cord white matter. The pathogenesis of myelin degeneration in this model has not been clarified, but morphologic and immunologic data suggest that the host immune response plays a major role in the production of myelin injury. Because of low virus titers in infected adult mice and of the small size of TMEV, virus particles have never been observed in this demyelinating model. Yet elucidation of the types of cells in the CNS supporting virus replication would be important for a better understanding of both virus persistence and virus-induced demyelinating pathology. The present paper is a sequential study of the localization of TMEV in the spinal cord in infected mice by ultrastructural immunohistochemical techniques. Results indicate that virus replication is mainly in neurons during the acute phase of the disease, while in the chronic phase viral inclusions are mainly found in macrophages in and around demyelinating lesions. Other cells are also infected, but to a lesser degree. In the neuronal system both axoplasmic and dendritic flow appear to facilitate the spread of virus in the CNS. In macrophages, the presence of virus particles and the association of virus with altered components of the cytoskeleton support active virus production rather than simple internalization. The macrophage appears to play an important role in both the establishment of virus persistence and in the process of demyelination in this animal model.

  11. Immunohistochemical expression of aberrant Notch-1 signaling in vitiligo: an implication for pathogenesis.

    PubMed

    Seleit, Iman; Bakry, Ola Ahmed; Abdou, Asmaa Gaber; Dawoud, Noha Mohammed

    2014-06-01

    The etiopathogenetic mechanisms leading to pigment loss in vitiligo are not fully understood. Notch signaling is required for development and maintenance of melanocyte lineage and acts as a key component among keratinocyte-melanocyte interactions. The current study aimed to investigate the possible role of Notch signaling and its effect on the whole melanocyte lineage in vitiligo and correlating it with the different clinicopathologic parameters. Using immunohistochemical technique, Notch-1 expression was evaluated in 50 lesional and 20 perilesional biopsies of patients with vitiligo in comparison with 20 normal skin biopsies as a control group. Lesional biopsies were stained with human melanoma black-45 and tyrosinase-related protein-2 to demonstrate the melanocyte lineage. Membranous and/or nuclear expression of Notch-1 was in favor of control and perilesional skin, whereas cytoplasmic expression appeared only in vitiliginous lesions (P < .05). Membranous and/or nuclear expression of Notch-1 was significantly associated with epidermal human melanoma black-45 positivity (P = .01) and percentage of expression in both epidermis (P = .02) and hair follicles (P = .03) of lesional skin. Cytoplasmic pattern of Notch-1 expression in epidermis was significantly found in lesions with white hair (P = .04) and in cases with marked keratinocyte vacuolization (P = .03). Segmental and acrofacial vitiligo were associated with mild to moderate Notch-1 intensity, whereas generalized vitiligo was associated with strong intensity of expression (P = .02). In conclusion, Notch-1 signaling is inactivated in vitiligo with consequent loss of epidermal and/or follicular active melanocytes. Aberrant Notch signaling in vitiliginous white hair and acral and segmental vitiligo may be the cause of their treatment resistance.

  12. Immunohistochemical determination of ETS-1 oncoprotein expression in urothelial carcinomas of the urinary bladder.

    PubMed

    Sari, Aysegul; Calli, Aylin; Gorgel, Sacit Nuri; Altinboga, Aysegul Aksoy; Kara, Cengiz; Dincel, Cetin; Cakalagaoglu, Fulya

    2012-03-01

    ETS-1 protooncogene is an important transcription factor that plays a role in the regulation of physiological processes, such as cell proliferation and differentiation. ETS-1 is thought to be related to the growth of carcinoma cells by its regulation of the transcription of matrix metalloproteinases and urokinase-type plasminogen activator. In this study, we aimed to investigate the expression pattern of ETS-1 oncoprotein in urothelial carcinomas of the urinary bladder and determine its relationship with histopathologic parameters, including tumor grade and stage. One hundred six specimens of urothelial carcinoma and a total of 14 normal urothelium were analyzed immunohistochemically with anti-ETS-1 monoclonal antibody. The normal urothelium showed positive ETS-1 immunostaining. ETS-1 expression remained high in low-grade and noninvasive tumors, whereas it frequently decreased in high-grade or invasive carcinomas. Interestingly, ETS-1 was highly expressed in the basal cell layer of the noninvasive urothelial carcinomas. ETS-1 expression showed a strong negative correlation with the tumor grade (P<0.001; r, -0.67) and stage (P<0.001; r, -0.75). The nonmuscle-invasive tumors (pTa+pT1) and noninvasive tumors (pTa) had significantly higher ETS-1 expression than the muscle-invasive tumors (pT2; P<0.001) and invasive tumors (pT1+pT2; P<0.001), respectively. Results of our study show that decreased ETS-1 expression is significantly associated with high grade and advanced stage in urothelial carcinomas of the urinary bladder, and that the downregulation of ETS-1 expression may be a marker of the aggressiveness of such malignancies.

  13. Cancer stem cells in hepatocellular carcinoma - an immunohistochemical study with histopathological association

    PubMed Central

    Matthai, Smita Mary; Ramakrishna, Banumathi

    2015-01-01

    Background & objectives: Cancer stem cells (CSCs) may be responsible for tumour recurrence and resistance to chemotherapy in hepatocellular carcinoma (HCC). This study was carried out to evaluate the association between histological parameters and liver CSCs (LCSC) in HCC, and to compare distribution of liver CSCs in HCC associated with and without hepatitis B virus (HBV) infection. Methods: Seventy nine tumours (49 surgical resections from 46 patients, and 30 from autopsy) were reviewed. Immunohistochemical staining for the LCSC marker EpCAM (epithelial cell adhesion molecule), liver progenitor cell (LPC) markers CK19 (cytokeratin 19) and neural cell adhesion molecule (NCAM) were performed and were associated with histological features of tumour behaviour. Results: Thirty three tumours (41.8%) showed positive staining for EpCAM. CK19 and NCAM expression were seen in 26 (32.9%) and four (5.1%) tumours, respectively. The expression of EpCAM and CK19 was significantly associated with each other (P<0.001). EpCAM expression was significantly associated with clinical and histological features indicating aggressive tumour behaviour, including younger age of onset, higher serum alpha foetoprotein (AFP) levels, tumour cell dedifferentiation, increased mitotic activity, and vascular invasiveness. There was no significant difference in expression of EpCAM, CK19 and NCAM between HBV positive and negative HCC. Interpretation & conclusions: The LCSC marker EpCAM was expressed in less than half of HCC, was independent of HBV aetiology, and was strongly associated with clinical and histological features of aggressive tumour behaviour. Positive staining for CK19 suggests a possible LPC origin of the EpCAM positive HCCs. PMID:26609030

  14. Physical activity and sedentary activity patterns among children and adolescents: a latent class analysis approach

    PubMed Central

    Heitzler, Carrie; Lytle, Leslie; Erickson, Darin; Sirard, John; Barr-Anderson, Daheia; Story, Marry

    2010-01-01

    Background While much is known about the overall levels of physical activity and sedentary activity among youth, few studies have attempted to define clusters of such behaviors. The purpose of this study was to identify and describe unique classes of youth based on their participation in a variety of physical activity and sedentary behaviors. Methods Latent class analysis was used to characterize segments of youth based on patterns of self-reported and accelerometer-measured participation in 12 behaviors. Children and adolescents (N=720) from 6th–11th grade were included in the analysis. Differences in class membership were examined using multinomial logistic regression. Results Three distinct classes emerged for boys and girls. Among boys, the three classes were characterized as: (1) “Active” (42.1%), (2) “Sedentary” (24.9%), and (3) “Low Media/Moderate Activity” (33.0%). For girls, classes were: (1) “Active” (18.7%), (2) “Sedentary” (47.6%), and (3) “Low Media/Functional Activity” (33.7%). Significant differences were found between the classes for a number of demographic indicators including the proportion in each class who were classified as overweight or obese. Conclusions The behavioral profiles of the classes identified in this study can be used to suggest possible audience segments for intervention and to tailor strategies appropriately. PMID:21597117

  15. Automated extraction of precise protein expression patterns in lymphoma by text mining abstracts of immunohistochemical studies

    PubMed Central

    Chang, Jia-Fu; Popescu, Mihail; Arthur, Gerald L.

    2013-01-01

    Background: In general, surgical pathology reviews report protein expression by tumors in a semi-quantitative manner, that is, -, -/+, +/-, +. At the same time, the experimental pathology literature provides multiple examples of precise expression levels determined by immunohistochemical (IHC) tissue examination of populations of tumors. Natural language processing (NLP) techniques enable the automated extraction of such information through text mining. We propose establishing a database linking quantitative protein expression levels with specific tumor classifications through NLP. Materials and Methods: Our method takes advantage of typical forms of representing experimental findings in terms of percentages of protein expression manifest by the tumor population under study. Characteristically, percentages are represented straightforwardly with the % symbol or as the number of positive findings of the total population. Such text is readily recognized using regular expressions and templates permitting extraction of sentences containing these forms for further analysis using grammatical structures and rule-based algorithms. Results: Our pilot study is limited to the extraction of such information related to lymphomas. We achieved a satisfactory level of retrieval as reflected in scores of 69.91% precision and 57.25% recall with an F-score of 62.95%. In addition, we demonstrate the utility of a web-based curation tool for confirming and correcting our findings. Conclusions: The experimental pathology literature represents a rich source of pathobiological information, which has been relatively underutilized. There has been a combinatorial explosion of knowledge within the pathology domain as represented by increasing numbers of immunophenotypes and disease subclassifications. NLP techniques support practical text mining techniques for extracting this knowledge and organizing it in forms appropriate for pathology decision support systems. PMID:23967385

  16. Bone response to biomimetic implants delivering BMP-2 and VEGF: an immunohistochemical study.

    PubMed

    Ramazanoglu, Mustafa; Lutz, Rainer; Rusche, Philipp; Trabzon, Levent; Kose, Gamze Torun; Prechtl, Christopher; Schlegel, Karl Andreas

    2013-12-01

    This animal study evaluated bone healing around titanium implant surfaces biomimetically coated with bone morphogenic protein-2 (BMP-2) and/or vascular endothelial growth factor (VEGF) by examining bone matrix proteins and mineralisation. Five different implant surfaces were established: acid-etched surface (AE), biomimetic calcium phosphate surface (CAP), BMP-2 loaded CAP surface, VEGF loaded CAP surface and dual BMP-2 + VEGF loaded CAP surface. The implants were inserted into calvariae of adult domestic pigs. For the comparison of osteoconductive capacity of each surface, bone mineral density and expression of bone matrix proteins (collagen I, BMP-2/4, osteocalcin and osteopontin) inside defined chambers around the implant were assessed using light microscopy and microradiography and immunohistochemical analysis at 1, 2 and 4 weeks. In the both groups delivering BMP-2, the bone mineral density was significantly enhanced after 2 weeks and the highest value was measured for the group BMP + VEGF. In the group VEGF, collagen I and BMP-2/4 expressions were significantly up-regulated at the first and second weeks. The percentage of BMP-2/4 positive cells in the group BMP + VEGF was significantly enhanced compared with the groups AE and CAP at the second week. Although the highest osteocalcin and osteopontin expression values were observed for the group BMP + VEGF after 2 weeks, no statistically significant difference in osteocalcin and osteopontin expressions was found between all groups at any time. It was concluded that combined delivery of BMP-2 and VEGF favoured bone mineralisation and expression of important bone matrix proteins that might explain synergistic interaction between both growth factors. PMID:23434516

  17. Bone response to biomimetic implants delivering BMP-2 and VEGF: an immunohistochemical study.

    PubMed

    Ramazanoglu, Mustafa; Lutz, Rainer; Rusche, Philipp; Trabzon, Levent; Kose, Gamze Torun; Prechtl, Christopher; Schlegel, Karl Andreas

    2013-12-01

    This animal study evaluated bone healing around titanium implant surfaces biomimetically coated with bone morphogenic protein-2 (BMP-2) and/or vascular endothelial growth factor (VEGF) by examining bone matrix proteins and mineralisation. Five different implant surfaces were established: acid-etched surface (AE), biomimetic calcium phosphate surface (CAP), BMP-2 loaded CAP surface, VEGF loaded CAP surface and dual BMP-2 + VEGF loaded CAP surface. The implants were inserted into calvariae of adult domestic pigs. For the comparison of osteoconductive capacity of each surface, bone mineral density and expression of bone matrix proteins (collagen I, BMP-2/4, osteocalcin and osteopontin) inside defined chambers around the implant were assessed using light microscopy and microradiography and immunohistochemical analysis at 1, 2 and 4 weeks. In the both groups delivering BMP-2, the bone mineral density was significantly enhanced after 2 weeks and the highest value was measured for the group BMP + VEGF. In the group VEGF, collagen I and BMP-2/4 expressions were significantly up-regulated at the first and second weeks. The percentage of BMP-2/4 positive cells in the group BMP + VEGF was significantly enhanced compared with the groups AE and CAP at the second week. Although the highest osteocalcin and osteopontin expression values were observed for the group BMP + VEGF after 2 weeks, no statistically significant difference in osteocalcin and osteopontin expressions was found between all groups at any time. It was concluded that combined delivery of BMP-2 and VEGF favoured bone mineralisation and expression of important bone matrix proteins that might explain synergistic interaction between both growth factors.

  18. Immunohistochemical and ultrastructural study of pituitary folliculostellate cells during aging in rats.

    PubMed

    Cónsole, G M; Jurado, S B; Riccillo, F L; Gómez Dumm, C L

    2000-01-01

    The impact of aging on pituitary folliculostellate (FS) cells is not well known. The aim of the work reported here was to carry out a quantitative immunohistochemical assessment of the FS population in male and female rats during aging and to correlate the findings with possible changes at the ultrastructural level. Young (4 months), old (20 months) and senescent (29 months) Sprague-Dawley rats of both sexes were sacrificed by rapid decapitation, their pituitaries dissected and processed by both light immunohistochemistry and electron microscopy. Serial sections (4 microm) were obtained at different levels and immunostained by means of rabbit anti-S100 serum as the primary antibody and a peroxidase-mediated EnVision System (Dako). Measurement of volume density (VD) and cell density (CD) was made in S100-reacting elements by means of an image analysis system (Imaging Technology, Optimas). These parameters were found to be significantly (p < 0.05) decreased in old and senescent rats as compared to young animals. In senescent females, which presented a high incidence of microprolactinomas, a significant (p < 0.01) increment of VD and CD was observed in FS cells in the area surrounding the adenomas, together with a marked decrease in those parameters within the tumors. Sexual dimorphism was not found except for the prolactinoma-bearing female group. The ultrastructure of FS cells showed the typical characteristics previously described in the pituitary gland. Only moderate changes in the endoplasmic reticulum were observed in old and senescent animals. We conclude that aging has a clear effect on the morphology of the pituitary FS cell population.

  19. The effects of antidepressants and pilocarpine on rat parotid glands: an immunohistochemical study

    PubMed Central

    Mattioli, Tatiana Maria Folador; da Silva, Silvana; Grégio, Ana Maria Trindade; Machado, Maria Ângela Naval; de Lima, Antônio Adilson Soares; Azevedo-Alanis, Luciana Reis

    2011-01-01

    OBJECTIVES: To evaluate the effects of antidepressants and pilocarpine on the quantity of myoepithelial cells and on the proliferation index of the epithelial cells of rat parotid glands. INTRODUCTION: Hyposalivation, xerostomia, and alterations in saliva composition are important clinical side effects related to the use of antidepressants. METHODS: Ninety male Wistar rats were allocated to nine groups. The control groups received saline for 30 (group C30) or 60 days (group C60) or pilocarpine for 60 days (group Pilo). The experimental groups were administered fluoxetine (group F30) or venlafaxine for 30 days (group V30); fluoxetine (group FS60) or venlafaxine (group VS60) with saline for 60 days; or fluoxetine (group FP60) or venlafaxine (group VP60) with pilocarpine for 60 days. Parotid gland specimens were processed, and the immunohistochemical expression of calponin and proliferating cell nuclear anti-antigen on the myoepithelial and parenchymal cells, respectively, was evaluated. Analysis of variance (ANOVA), Tukey HSD and Games-Howell tests were applied to detect differences among groups (p<0.05). RESULTS: Compared with the controls, chronic exposure to antidepressants was associated with an increase in the number of positively stained cells for calponin. In addition, venlafaxine administration for 30 days was associated with an increase in the number of positively stained cells for proliferating cell nuclear anti-antigen. Fluoxetine and pilocarpine (group FP60) induced a significant decrease in the number of positively stained cells for calponin compared with all other groups. CONCLUSIONS: The number of positively stained cells for calponin increased after chronic administration of antidepressants. The proliferation index of the epithelial cells of rat parotid glands was not altered by the use of antidepressants for 60 days. PMID:22179167

  20. PLAG1 expression in cutaneous mixed tumors: an immunohistochemical and molecular genetic study.

    PubMed

    Matsuyama, Atsuji; Hisaoka, Masanori; Hashimoto, Hiroshi

    2011-11-01

    Cutaneous mixed tumors, also known as chondroid syringomas, are benign cutaneous adnexal tumors that exhibit remarkable histopathological similarities to pleomorphic adenoma of the salivary gland. Thus far, there is little information on the genetic profiles of cutaneous mixed tumors, although specific genetic aberrations including fusion genes involving PLAG1 and HMGA2 have been demonstrated in pleomorphic adenomas. In the present study, we conducted an immunohistochemical evaluation of PLAG1 and a reverse transcription-polymerase chain reaction (RT-PCR) assay to detect fusion gene transcripts associated with pleomorphic adenoma, including the CTNNB1-PLAG1, LIFR-PLAG1, CHCHD7-PLAG1, TCEA1-PLAG1, HMGA2-FHIT, HMGA2-NFIB, and HMGA2-WIF1 fusion transcripts; this was performed using formalin-fixed paraffin-embedded tumor tissue specimens of 16 cutaneous mixed tumors including one sample with an adenocarcinoma component. All 16 cutaneous mixed tumors were immunoreactive to PLAG1, which was predominantly expressed in cells with myoepithelial or chondroid differentiation accounting for >80% of cells, whereas PLAG1 expression in glandular or squamous tumor cells was restricted to <20% of cells. The carcinoma component in the mixed tumor was also positive for PLAG1. On the other hand, all eight cutaneous adnexal tumors other than the mixed tumor were negative for PLAG1. In RT-PCR analysis, no fusion gene transcripts involving PLAG1 or HMGA2 were identified in any of the cases. Concordant with previous studies, our results support the close relationship between cutaneous mixed tumors and pleomorphic adenomas of the salivary gland. However, the mechanism of PLAG1 expression in cutaneous mixed tumors appears to be possibly different from that of pleomorphic adenomas.

  1. Characterization of indoor cooking aerosol using neutron activation analysis

    SciTech Connect

    Wu, D.; Landsberger, S.; Larson, S. )

    1993-01-01

    Suspended particles in air are potentially harmful to human health, depending on their sizes and chemical composition. Residential indoor particles mainly come from (a) outdoor sources that are transported indoors, (b) indoor dust that is resuspended, and (c) indoor combustion sources, which include cigarette smoking, cooking, and heating. Jedrychowski stated that chronic phlegm in elderly women was strongly related to the cooking exposure. Kamens et al. indicated that cooking could generate small particles (<0.1 [mu]m), and cooking one meal could contribute [approximately]5 to 18% of total daytime particle volume exposure. Although cooking is a basic human activity, there are not many data available on the properties of particles generated by this activity. Some cooking methods, such as stir-frying and frying, which are the most favored for Chinese and other Far East people, generate a large quantity of aerosols. This research included the following efforts: 1. investigating particle number concentrations, distributions, and their variations with four different cooking methods and ventilation conditions; 2. measuring the chemical composition of cooking aerosol samples by instrumental neutron activation analysis.

  2. The role of neutron activation analysis in nutritional biomonitoring programs

    SciTech Connect

    Iyengar, V.

    1988-01-01

    Nutritional biomonitoring is a multidisciplinary task and an integral part of a more general bioenvironmental surveillance. In its comprehensive form, it is a combination of biological, environmental, and nutrient monitoring activities. Nutrient monitoring evaluates the input of essential nutrients required to maintain vital bodily functions; this includes vigilance over extreme fluctuations of nutrient intake in relation to the recommended dietary allowances and estimated safe and adequate daily dietary intakes and adherence to the goals of provisional tolerance limits. Environmental monitoring assesses the external human exposure via ambient pathways, namely, air, water, soil, food, etc. Biological monitoring quantifies a toxic agent and its metabolites in representative biologic specimens of an exposed organ to identify health effects. In practice, coordinating all three components of a nutritional biomonitoring program is complex, expensive, and tedious. Experience gained from the US National Health and Nutrition Examination Surveys demonstrates the problems involved. By far the most critical challenge faced here is the question of analytical quality control, particularly when trace element determinations are involved. Yet, measures to ensure reliability of analytical data are mandatory, and there are no short-cuts to this requirement. The purpose of this presentation is to elucidate the potential of neutron activation analysis (NAA) in nutritional biomonitoring activities.

  3. Dictionary learning and sparse recovery for electrodermal activity analysis

    NASA Astrophysics Data System (ADS)

    Kelsey, Malia; Dallal, Ahmed; Eldeeb, Safaa; Akcakaya, Murat; Kleckner, Ian; Gerard, Christophe; Quigley, Karen S.; Goodwin, Matthew S.

    2016-05-01

    Measures of electrodermal activity (EDA) have advanced research in a wide variety of areas including psychophysiology; however, the majority of this research is typically undertaken in laboratory settings. To extend the ecological validity of laboratory assessments, researchers are taking advantage of advances in wireless biosensors to gather EDA data in ambulatory settings, such as in school classrooms. While measuring EDA in naturalistic contexts may enhance ecological validity, it also introduces analytical challenges that current techniques cannot address. One limitation is the limited efficiency and automation of analysis techniques. M