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Sample records for subjects immunohistochemical staining

  1. Comparison of immunohistochemical and modified Giemsa stains ...

    African Journals Online (AJOL)

    BackgroundModified Giemsa staining has been favoured by many researchers because it is easy to perform but, like many other stains, demonstration of the bacteria depends on its morphology. It has been arged in some research circles that some of the organisms in the gastric mucosa may not be true H.pylori.

  2. Immunohistochemical CD3 staining detects additional patients with celiac disease.

    Science.gov (United States)

    Mubarak, Amani; Wolters, Victorien M; Houwen, Roderick H J; ten Kate, Fiebo J W

    2015-06-28

    To investigate whether performing immunohistochemical CD3 staining, in order to improve the detection of intra-epithelial lymphocytosis, has an additional value in the histological diagnosis of celiac disease. Biopsies obtained from 159 children were stained by hematoxylin and eosin (HE) and evaluated using the Marsh classification. CD3 staining was subsequently evaluated separately and independently. Differences in evaluation between the routine HE sections and CD3 staining were present in 20 (12.6%) cases. In 10 (6.3%) patients the diagnosis of celiac disease (Marsh II and III) changed on examination of CD3 staining: in 9 cases, celiac disease had initially been missed on the HE sections, while 1 patient had been over-diagnosed on the routine sections. In all patients, the final diagnosis based on CD3 staining, was concordant with serological results, which was not found previously. In the other 10 (12.3%) patients, the detection of sole intra-epithelial lymphocytosis (Marsh I) improved. Nine patients were found to have Marsh I on CD3 sections, which had been missed on routine sections. Interestingly, the only patient with negative serology had Giardiasis. Finally, in 1 patient with negative serology, in whom Marsh I was suspected on HE sections, this diagnosis was withdrawn after evaluation of the CD3 sections. Staining for CD3 has an additional value in the histological detection of celiac disease lesions, and CD3 staining should be performed when there is a discrepancy between serology and the diagnosis made on HE sections.

  3. CD3 immunohistochemical staining in diagnosis of lymphocytic colitis.

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    Fiehn, Anne-Marie Kanstrup; Engel, Ulla; Holck, Susanne; Munck, Lars Kristian; Engel, Peter Johan Heiberg

    2016-02-01

    Microscopic colitis (MC) is a common cause of chronic watery diarrhea. Traditionally, MC encompasses the 2 subgroups lymphocytic colitis (LC) and collagenous colitis, but recently, an additional subgroup, MC incomplete, has been introduced. Distinguishing between the subgroups relies exclusively on histopathologic evaluation. In the present study, 4 pathologists evaluated 156 archived biopsies originally diagnosed as LC or LC incomplete (LCi). Each pathologist assigned a diagnosis of LC, LCi, or nonspecific inflammation to all cases at 2 independent assessments. At the first assessment, hematoxylin and eosin (HE) stainings were available. At the second assessment, a supplementary CD3 immunohistochemical staining was also available. The aim was to evaluate whether a supplementary CD3 would increase the diagnostic agreement among pathologists, and whether a CD3 stain would change the diagnosis based on HE staining only. After the complete assessment, the cases were divided into 3 groups, that is, full agreement, partial agreement, and disagreement. The CD3 staining increased the number of cases with full agreement from 60 to 78. One hundred thirty-one cases with agreement or partial diagnostic agreement based on HE + CD3 were compared with the HE diagnoses. In 44 (34%) of 131 cases, CD3 changed the diagnosis. Cases assigned to the LCi category based on HE were often changed by a supplementary CD3. Conclusively, it is recommended to use a CD3 before giving the histopathologic diagnosis of LCi. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. Microfluidics for rapid cytokeratin immunohistochemical staining in frozen sections.

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    Brajkovic, Saska; Dupouy, Diego G; de Leval, Laurence; Gijs, Martin Am

    2017-08-01

    Frozen sections (FS) of tumor samples represent a cornerstone of pathological intraoperative consultation and have an important role in the microscopic analysis of specimens during surgery. So far, immunohistochemical (IHC) stainings on FS have been demonstrated for a few markers using manual methods. Microfluidic technologies have proven to bring substantial improvement in many fields of diagnostics, though only a few microfluidic devices have been designed to improve the performance of IHC assays. In this work, we show optimization of a complete pan-cytokeratin chromogenic immunostaining protocol on FS using a microfluidic tissue processor into a protocol taking <12 min. Our results showed specificity and low levels of background. The dimensions of the microfluidic prototype device are compatible with the space constraints of an intraoperative pathology laboratory. We therefore anticipate that the adoption of microfluidic technologies in the field of surgical pathology can significantly improve the way FSs influence surgical procedures.

  5. p53 immunohistochemical staining patterns in oral squamous cell carcinoma

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    G Dundy

    2016-09-01

    Full Text Available Background: Mutation of p53 gene is one of the most common events in oral carcinogenesis. Accumulation of p53 protein has also been detected in premalignant lesions.Materials and Methods:  This study included 40 biopsy samples, which were received in department of pathology, Sarojini Naidu Medical College, Agra, to ascertain p53 expression by immunohistochemically, in patients with oral squamous cell carcinomas and to correlate its expression with histological grade, different sites in oral cavity and tobacco intake/smoking habits.Results: Out of 40 biopsies of oral mucosa, 03 showed normal oral mucosa and 37 were diagnosed as squamous cell carcinoma (SCC, most patients were in 5th and 6th decade and majority (86.5% of oral SCC were males with buccal mucosa being the most common site. There was a statistically significant difference in p53 expression between oral SCC and normal oral mucosa (p value <0.05. Of total 37 cases, 12 cases were well differentiated type, 16 moderately differentiated and 09 of poorly differentiated type of SCC. In each category, about two thirds were positive for p53 staining. Out of total 37 cases of oral SCC, 64.9% were positive and 35.1% were negative for p53 expression, 34 cases had positive history of tobacco intake/smoking habits, of which 23 cases were positive while 11 cases were negative for p53 staining.Conclusion: Abnormal p53 protein was detected in 64.9% of oral squamous cell carcinoma, but not in normal oral mucosa. p53 expression was associated with malignant transformation of oral mucosa. 

  6. Heterogeneity in Li-Fraumeni families: p53 mutation analysis and immunohistochemical staining.

    OpenAIRE

    MacGeoch, C; Turner, G; Bobrow, L G; Barnes, D M; Bishop, D T; Spurr, N K

    1995-01-01

    We have screened two families for constitutional TP53 mutations, one family with Li-Fraumeni syndrome and the other with features of this syndrome. We report a germline mutation in exon 7 of the TP53 gene in the family with "Li-Fraumeni-like" syndrome. The mutation occurred at codon 245 and causes a Gly-Ser amino acid change. It was inherited by both affected and unaffected subjects. Malignant tumours from all members of this family showed strong positive nuclear immunohistochemical staining ...

  7. Quantitative comparison of immunohistochemical staining measured by digital image analysis versus pathologist visual scoring

    Directory of Open Access Journals (Sweden)

    Rizzardi Anthony E

    2012-06-01

    Full Text Available Abstract Immunohistochemical (IHC assays performed on formalin-fixed paraffin-embedded (FFPE tissue sections traditionally have been semi-quantified by pathologist visual scoring of staining. IHC is useful for validating biomarkers discovered through genomics methods as large clinical repositories of FFPE specimens support the construction of tissue microarrays (TMAs for high throughput studies. Due to the ubiquitous availability of IHC techniques in clinical laboratories, validated IHC biomarkers may be translated readily into clinical use. However, the method of pathologist semi-quantification is costly, inherently subjective, and produces ordinal rather than continuous variable data. Computer-aided analysis of digitized whole slide images may overcome these limitations. Using TMAs representing 215 ovarian serous carcinoma specimens stained for S100A1, we assessed the degree to which data obtained using computer-aided methods correlated with data obtained by pathologist visual scoring. To evaluate computer-aided image classification, IHC staining within pathologist annotated and software-classified areas of carcinoma were compared for each case. Two metrics for IHC staining were used: the percentage of carcinoma with S100A1 staining (%Pos, and the product of the staining intensity (optical density [OD] of staining multiplied by the percentage of carcinoma with S100A1 staining (OD*%Pos. A comparison of the IHC staining data obtained from manual annotations and software-derived annotations showed strong agreement, indicating that software efficiently classifies carcinomatous areas within IHC slide images. Comparisons of IHC intensity data derived using pixel analysis software versus pathologist visual scoring demonstrated high Spearman correlations of 0.88 for %Pos (p  Virtual slides The virtual slide(s for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1649068103671302

  8. Heterogeneity in Li-Fraumeni families: p53 mutation analysis and immunohistochemical staining.

    Science.gov (United States)

    MacGeoch, C; Turner, G; Bobrow, L G; Barnes, D M; Bishop, D T; Spurr, N K

    1995-03-01

    We have screened two families for constitutional TP53 mutations, one family with Li-Fraumeni syndrome and the other with features of this syndrome. We report a germline mutation in exon 7 of the TP53 gene in the family with "Li-Fraumeni-like" syndrome. The mutation occurred at codon 245 and causes a Gly-Ser amino acid change. It was inherited by both affected and unaffected subjects. Malignant tumours from all members of this family showed strong positive nuclear immunohistochemical staining with antibodies CM-1 and DO1, directed against TP53. In contrast, no constitutional TP53 mutations were found in a "classic" Li-Fraumeni family. In this family positive staining was seen in both malignant and normal tissues. These results support previous findings that variants of the Li-Fraumeni syndrome exist since not all LFS families carry TP53 germline mutations. Secondly, immunohistochemical positivity is not synonymous with an underlying mutation and is therefore inadequate as an exclusive diagnostic marker.

  9. Quantitative comparison of immunohistochemical staining measured by digital image analysis versus pathologist visual scoring

    Science.gov (United States)

    2012-01-01

    Abstract Immunohistochemical (IHC) assays performed on formalin-fixed paraffin-embedded (FFPE) tissue sections traditionally have been semi-quantified by pathologist visual scoring of staining. IHC is useful for validating biomarkers discovered through genomics methods as large clinical repositories of FFPE specimens support the construction of tissue microarrays (TMAs) for high throughput studies. Due to the ubiquitous availability of IHC techniques in clinical laboratories, validated IHC biomarkers may be translated readily into clinical use. However, the method of pathologist semi-quantification is costly, inherently subjective, and produces ordinal rather than continuous variable data. Computer-aided analysis of digitized whole slide images may overcome these limitations. Using TMAs representing 215 ovarian serous carcinoma specimens stained for S100A1, we assessed the degree to which data obtained using computer-aided methods correlated with data obtained by pathologist visual scoring. To evaluate computer-aided image classification, IHC staining within pathologist annotated and software-classified areas of carcinoma were compared for each case. Two metrics for IHC staining were used: the percentage of carcinoma with S100A1 staining (%Pos), and the product of the staining intensity (optical density [OD] of staining) multiplied by the percentage of carcinoma with S100A1 staining (OD*%Pos). A comparison of the IHC staining data obtained from manual annotations and software-derived annotations showed strong agreement, indicating that software efficiently classifies carcinomatous areas within IHC slide images. Comparisons of IHC intensity data derived using pixel analysis software versus pathologist visual scoring demonstrated high Spearman correlations of 0.88 for %Pos (p < 0.0001) and 0.90 for OD*%Pos (p < 0.0001). This study demonstrated that computer-aided methods to classify image areas of interest (e.g., carcinomatous areas of tissue specimens) and

  10. Segmentation of HER2 protein overexpression in immunohistochemically stained breast cancer images using Support Vector Machines

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    Pezoa, Raquel; Salinas, Luis; Torres, Claudio; Härtel, Steffen; Maureira-Fredes, Cristián; Arce, Paola

    2016-10-01

    Breast cancer is one of the most common cancers in women worldwide. Patient therapy is widely supported by analysis of immunohistochemically (IHC) stained tissue sections. In particular, the analysis of HER2 overexpression by immunohistochemistry helps to determine when patients are suitable to HER2-targeted treatment. Computational HER2 overexpression analysis is still an open problem and a challenging task principally because of the variability of immunohistochemistry tissue samples and the subjectivity of the specialists to assess the samples. In addition, the immunohistochemistry process can produce diverse artifacts that difficult the HER2 overexpression assessment. In this paper we study the segmentation of HER2 overexpression in IHC stained breast cancer tissue images using a support vector machine (SVM) classifier. We asses the SVM performance using diverse color and texture pixel-level features including the RGB, CMYK, HSV, CIE L*a*b* color spaces, color deconvolution filter and Haralick features. We measure classification performance for three datasets containing a total of 153 IHC images that were previously labeled by a pathologist.

  11. Differentiating neurotized melanocytic nevi from neurofibromas using Melan-A (MART-1) immunohistochemical stain.

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    Chen, Yumei; Klonowski, Paul W; Lind, Anne C; Lu, Dongsi

    2012-07-01

    Neurotized melanocytic nevi and neurofibromas are common, benign cutaneous neoplasms. Usually they are histologically distinct from each other; however, neurotized melanocytic nevi and neurofibromas can be clinically and histologically similar. To determine whether Melan-A (MART-1) immunohistochemical stain is sufficient to differentiate neurotized melanocytic nevi from neurofibromas. Forty-nine consecutive specimens of melanocytic nevi with neurotization and 49 specimens of neurofibromas were selected. We used antibodies against Melan-A, S100, and neurofilament protein. All of the melanocytic nevi showed Melan-A staining within the neurotized areas, with most of the areas staining strongly positive, whereas all the neurofibromas were completely absent of Melan-A stain. All of the nevi, including the neurotized areas, stained strongly and diffusely for S100, whereas all the neurofibromas showed a distinctive, sharp, wavy pattern of S100 staining. Neurofilament protein showed scattered staining of both melanocytic nevi and neurofibromas. Our data indicate that Melan-A immunohistochemical staining is helpful in differentiating neurotized melanocytic nevi from neurofibromas when distinction on histomorphology alone is difficult.

  12. A standard tissue as a control for histochemical and immunohistochemical staining

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    Otali, D; Fredenburgh, J; Oelschlager, DK; Grizzle, WE

    2017-01-01

    The variable quality of histochemical and immunohistochemical staining of tissues may be attributed to pre-analytical and analytical variables. Both categories of variables frequently are undefined or inadequately controlled during specimen collection and preparation. Pre-analytical variables may alter the molecular composition of tissues, which results in variable staining; such variations may cause problems when different tissues are used as staining controls. We developed a standard tissue for use as a staining control. Our standard tissue contains five components: 1) nine combined human cell lines mixed with stroma from human spleen; 2) a squamous cancer cell line, A431; 3) fungus; 4) transverse sections of the mosquitofish and 5) normal human spleen. The first three components were embedded in HistoGel™ and all components were processed to paraffin and used to construct a single standard paraffin block. The muscles of mosquitofish and arteries of the spleen are positive controls for eosin staining, while other tissues are useful for assessing hematoxylin staining. The mosquitofish tissues also are excellent controls for the Masson trichrome stain and all mucin-related histochemical stains that we tested. The goblet cells of the intestine and skin stained strongly with Alcian blue, pH 2.5 (AB-2.5), mucicarmine, colloidal iron, periodic acid Schiff (PAS) or PAS-hematoxylin (PASH) and combination stains such as colloidal iron-PASH. Cell lines were not useful for evaluating histochemical stains except for PASH. The splenic stroma was a useful control for AB-2.5; however, eosin and mucin stains stained cell lines poorly, probably due to their rapid growth and associated loss of some differentiated characteristics such as production of mucins. Nevertheless, the cell lines were a critical control for immunohistochemical stains. Immunostaining of specific cell lines was consistent with the presence of markers, e.g., EGFr in DU145 cells. The cell lines expressed a

  13. Classification of feline intraocular neoplasms based on morphology, histochemical staining, and immunohistochemical labeling.

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    Grahn, Bruce H; Peiffer, Robert L; Cullen, Cheryl L; Haines, Deborah M

    2006-01-01

    The objectives of this study were to evaluate morphologic, histochemical, and immunohistochemical characteristics of well-differentiated and anaplastic intraocular neoplasms of cats, and to develop a diagnostic algorithm for, and investigate the association of ruptured lenses with these neoplasms. Seventy-five feline globes with intraocular neoplasms were stained with hematoxylin and eosin and examined by light microscopy. Morphologic diagnoses included 33 intraocular sarcomas, 17 diffuse iris melanomas, 15 lymphosarcomas, three ciliary adenomas, one metastatic carcinoma, and six undifferentiated intraocular neoplasms. Sections of these globes were then stained with periodic acid Schiff (PAS), and immunohistochemical (IHC) labels for various cellular markers. Histochemical staining and IHC labeling confirmed cellular differentiation in 73/75 neoplasms but was discordant with morphologic diagnoses in 8/75. These included four neoplasms morphologically diagnosed as lymphosarcomas but which expressed differentiation antigens consistent with melanoma (n = 3) or ciliary adenocarcinoma (n = 1), and four tumors morphologically diagnosed as intraocular sarcomas that expressed differentiation antigens for melanoma (n = 2), metastatic carcinoma (n = 1), or remained undifferentiated (n = 1). Immunohistochemical labeling suggested a diagnosis in 5/6 morphologically undifferentiated neoplasms including one intraocular sarcoma, two diffuse iridal melanomas, and two ciliary adenocarcinomas. Based upon morphologic, histochemical, and IHC characterization, ruptured lens capsules were detected in 28/30 intraocular sarcomas, 3/24 diffuse iris melanomas and 1/11 lymphosarcomas, but not in ciliary epithelial neoplasms, metastatic carcinomas, or undifferentiated intraocular neoplasms. An algorithm is provided that facilitates stain and IHC label selection for differentiating anaplastic intraocular feline neoplasms.

  14. Double immunohistochemical staining with laminin 5 (γ2 chain) and collagen IV in colorectal neoplasms

    DEFF Research Database (Denmark)

    Fiehn, Anne-Marie Kanstrup; Bzorek, Michael; Warnecke, Mads

    2016-01-01

    Colorectal cancer (CRC) is one of the most common cancer diagnoses in the Western world. It is outnumbered several times by the precursor stage adenoma. The aim of this study was to describe the expression pattern with a double immunohistochemical staining for laminin 5 (γ2) and collagen IV...... in different colorectal neoplasms. This might be a supplementary tool to morphology in diagnostic dilemmas as microinvasive pT1 tumors and adenomas with pseudoinvasion. Laminin 5 has been shown to stain in invasive tumor cells, while collagen IV highlights the basement membrane (BM). Fifty-seven patients...... intact around normal crypts. In invasive tumors, laminin 5 stained intensely, and the BM was absent or focally discontinuous. The expression in adenomas and in pseudoinvasive areas was less consistent. The study suggests that double immunostaining with collagen IV and laminin 5 might be useful...

  15. Identification of fungi in fungal ball sinusitis: comparison between MUC5B immunohistochemical and Grocott methenamine silver staining.

    Science.gov (United States)

    Ma, Ling; Xu, Rui; Shi, Jianbo; Zhou, Wei; Xu, Geng; Jiang, Guangli; Li, Guangqi; Chen, Zhiqiang

    2013-11-01

    MUC5B immunohistochemical staining was a valuable method for identifying fungi in fungal ball sinusitis, especially in distinguishing aspergillus and mucor. Combined infection with Aspergillus and Mucor fungi was the most common pattern in fungal ball sinusitis. To assess the value of MUC5B immunohistochemical staining in identifying fungi in fungal ball sinusitis by comparing it with conventional Grocott methenamine silver (GMS) staining. GMS staining and MUC5B immunohistochemical staining were used to identify fungi in mucopurulent cheesy or clay-like tissues from sinuses in 180 fungal ball sinusitis patients, and the examination results were compared. In 180 samples of fungal ball sinusitis, GMS staining showed Aspergillus in 130, Mucor in 88, and Candida albicans in 6, while MUC5B immunohistochemical staining identified Aspergillus in 166, Mucor in 172, and Candida albicans in 16. The fungal detection rate for MUC5B immunohistochemical staining was markedly greater than that for GMS staining (p sinusitis. Mixed infection of Aspergillus and Mucor was present in 146 of 180 patients (81.1%).

  16. Evaluation of Decalcification Techniques for Rat Femurs Using HE and Immunohistochemical Staining

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    Haixia Liu

    2017-01-01

    Full Text Available Aim. In routine histopathology, decalcification is an essential step for mineralized tissues. The purpose of this study is to evaluate the effects of different decalcification solutions on the morphological and antigenicity preservation in Sprague Dawley (SD rat femurs. Materials and Methods. Four different decalcification solutions were employed to remove the mineral substances from rat femurs, including 10% neutral buffered EDTA, 3% nitric acid, 5% nitric acid, and 8% hydrochloric acid/formic acid. Shaking and low temperature were used to process the samples. The stainings of hematoxylin-eosin (HE and immunohistochemical (IHC were employed to evaluate the bone morphology and antigenicity. Key Findings. Different decalcification solutions may affect the quality of morphology and the staining of paraffin-embedded sections in pathological examinations. Among four decalcifying solutions, 3% nitric acid is the best decalcifying agent for HE staining. 10% neutral buffered EDTA and 5% nitric acid are the preferred decalcifying agents for IHC staining. Significance. The current study investigated the effects of different decalcifying agents on the preservation of the bone structure and antigenicity, which will help to develop suitable protocols for the analyses of the bony tissue.

  17. Automatic Cell Segmentation Using a Shape-Classification Model in Immunohistochemically Stained Cytological Images

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    Shah, Shishir

    This paper presents a segmentation method for detecting cells in immunohistochemically stained cytological images. A two-phase approach to segmentation is used where an unsupervised clustering approach coupled with cluster merging based on a fitness function is used as the first phase to obtain a first approximation of the cell locations. A joint segmentation-classification approach incorporating ellipse as a shape model is used as the second phase to detect the final cell contour. The segmentation model estimates a multivariate density function of low-level image features from training samples and uses it as a measure of how likely each image pixel is to be a cell. This estimate is constrained by the zero level set, which is obtained as a solution to an implicit representation of an ellipse. Results of segmentation are presented and compared to ground truth measurements.

  18. Immunohistochemical/histochemical double staining method in the study of the columnar metaplasia of the oesophagus

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    D. Cabibi

    2014-03-01

    Full Text Available Intestinal metaplasia in Barrett’s oesophagus (BO represents an important risk factor for oesophageal adenocarcinoma. Instead, few and controversial data are reported about the progression risk of columnar-lined oesophagus without intestinal metaplasia (CLO, posing an issue about its clinical management. The aim was to evaluate if some immunophenotypic changes were present in CLO independently of the presence of the goblet cells. We studied a series of oesophageal biopsies from patients with endoscopic finding of columnar metaplasia, by performing some immunohistochemical stainings (CK7, p53, AuroraA combined with histochemistry (Alcian-blue and Alcian/PAS, with the aim of simultaneously assess the histochemical features in cells that shows an aberrant expression of such antigens. We evidenced a cytoplasmic expression of CK7 and a nuclear expression of Aurora A and p53,  both in goblet cells of BO and in non-goblet cells of CLO, some of which showing mild dysplasia. These findings suggest that some immunophenotypic changes are present in CLO and they can precede the appearance of the goblet cells or can be present independently of them, confirming the conception of BO as the condition characterized by any extention of columnar epithelium. This is the first study in which a combined immunohistochemical/histochemical method has been applied to Barrett pathology.

  19. Tumor Budding Detection by Immunohistochemical Staining is Not Superior to Hematoxylin and Eosin Staining for Predicting Lymph Node Metastasis in pT1 Colorectal Cancer.

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    Okamura, Takuma; Shimada, Yoshifumi; Nogami, Hitoshi; Kameyama, Hitoshi; Kobayashi, Takashi; Kosugi, Shin-ichi; Wakai, Toshifumi; Ajioka, Yoichi

    2016-05-01

    Tumor budding is recognized as an important risk factor for lymph node metastasis in pT1 colorectal cancer. Immunohistochemical staining for cytokeratin has the potential to improve the objective diagnosis of tumor budding over detection based on hematoxylin and eosin staining. However, it remains unclear whether tumor budding detected by immunohistochemical staining is a significant predictor of lymph node metastasis in pT1 colorectal cancer. The purpose of this study was to clarify the clinical significance of tumor budding detected by immunohistochemical staining in comparison with that detected by hematoxylin and eosin staining. This was a retrospective study. The study was conducted at Niigata University Medical & Dental Hospital. We enrolled 265 patients with pT1 colorectal cancer who underwent surgery with lymph node dissection. Tumor budding was evaluated by both hematoxylin and eosin and immunohistochemical staining with the use of CAM5.2 antibody. Receiver operating characteristic curve analyses were conducted to determine the optimal cutoff values for tumor budding detected by hematoxylin and eosin and CAM5.2 staining. Univariate and multivariate analyses were performed to identify the significant factors for predicting lymph node metastasis. Receiver operating characteristic curve analyses revealed that the cutoff values for tumor budding detected by hematoxylin and eosin and CAM5.2 staining for predicting lymph node metastases were 5 and 8. On multivariate analysis, histopathological differentiation (OR, 6.21; 95% CI, 1.16-33.33; p = 0.03) and tumor budding detected by hematoxylin and eosin staining (OR, 4.91; 95% CI, 1.64-14.66; p = 0.004) were significant predictors for lymph node metastasis; however, tumor budding detected by CAM5.2 staining was not a significant predictor. This study was limited by potential selection bias because surgically resected specimens were collected instead of endoscopically resected specimens. Tumor budding detected by

  20. Enhanced antigen detection in immunohistochemical staining using a 'digitized' chimeric antibody.

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    Eng, Hui-Yan; Wang, Cheng-I; Xue, Yuezhen; Lee, Chia-Yin; Zulkifli, Sarah Binte; Chiam, Poh-Cheang; Ghadessy, Farid J; Lane, David P

    2016-01-01

    The immunohistochemical (IHC) staining of mouse tissue sections using antibodies of mouse origin can result in high nonspecific background due to the staining of endogenous immunoglobulins (Igs) by enzyme-conjugated secondary antibodies. In order to obviate this issue, we developed a chimeric mouse-human anti-p53 monoclonal antibody (MH242) by grafting the variable regions of a known mouse antibody into a human Ig scaffold. This facilitated use of an anti-human secondary antibody, and resulted in near-zero background when compared with its parental mouse monoclonal antibody (PAb242). Furthermore, the chimeric antibody enabled reproducible detection of mutant p53 (homozygous R172H) expression in mouse tissue, an observation hitherto largely equivocal based on the use of existing antibodies. The approach we describe leads to the generation of tractable antibody reagents, whose integrity can be readily verified through DNA sequencing of expressor plasmids. The wide-spread adoption of such 'digitized' antibodies should reduce experimental disparities that can commonly arise through variations in antibody quality. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  1. Sebaceous gland carcinoma of the ocular adnexa - variability in clinical and histological appearance with analysis of immunohistochemical staining patterns.

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    Schmitz, Eva Janine; Herwig-Carl, Martina C; Holz, Frank G; Loeffler, Karin U

    2017-11-01

    The purpose of the study was to evaluate the characteristics of sebaceous gland carcinoma (SGC) of the ocular adnexae, which is due to a high variability in clinical, histological and immunohistochemical characteristics often challenging to diagnose. Records of six patients with SGC were reviewed, who underwent surgical excision and who were histologically diagnosed with SGC. For comparison, there were specimens from four patients with basal cell carcinoma (BCC) and four patients with squamous cell carcinoma (SCC). Histological and immunohistochemical analysis included stains for HE, cytokeratins (CKpan, Cam5.2), epithelial membrane antigen (EMA), androgen receptor (AR441), perforin and adipophilin. SGC's were located in the upper (n = 2) or lower (n = 4) eyelid and were associated with various presenting clinical signs including chalazion-like lesions with pyogenic granuloma (n = 1), papillomatous conjunctival tumors (n = 3), a hyperkeratotic exophytic neoplasm (n = 1) and an ulcerating crusted lesion resembling chronic blepharitis (n = 1). The treatment was tumor resection, followed (if necessary) by adjuvant therapy with topical Mitomycin C (n = 2). Histologic characteristics included basophilic pleomorphic cells with vacuolated cytoplasm, prominent nucleoli, mitotic figures and in some cases pagetoid spread (n = 2). CKpan, EMA and Cam5.2 showed strong positive immunoreactivity in all specimens (SGC, BCC, SCC). Perforin immunostaining showed a varying, but overall weak, non-specific cytoplasmatic staining reaction in all lesions. AR441 positivity was noted with variable intensity in almost all lesions and in particular in pagetoid spread in contrast to non-tumor cells. Adipophilin showed an annular staining of lipid granules in immature sebaceous cells in SGC in contrast to a more granular staining pattern in BCC and SCC. SGCs display a variety of clinical signs and may mimic many other lesions. Tumor resection, followed by histological and

  2. Proposals for best-quality immunohistochemical staining of paraffin-embedded brain tissue slides in forensics.

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    Trautz, Florian; Dreßler, Jan; Stassart, Ruth; Müller, Wolf; Ondruschka, Benjamin

    2018-01-03

    Immunohistochemistry (IHC) has become an integral part in forensic histopathology over the last decades. However, the underlying methods for IHC vary greatly depending on the institution, creating a lack of comparability. The aim of this study was to assess the optimal approach for different technical aspects of IHC, in order to improve and standardize this procedure. Therefore, qualitative results from manual and automatic IHC staining of brain samples were compared, as well as potential differences in suitability of common IHC glass slides. Further, possibilities of image digitalization and connected issues were investigated. In our study, automatic staining showed more consistent staining results, compared to manual staining procedures. Digitalization and digital post-processing facilitated direct analysis and analysis for reproducibility considerably. No differences were found for different commercially available microscopic glass slides regarding suitability of IHC brain researches, but a certain rate of tissue loss should be expected during the staining process.

  3. Application of immunohistochemical staining to detect antigen destruction as a measure of tissue damage.

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    Onul, Abdullah; Colvard, Michael D; Paradise, William A; Elseth, Kim M; Vesper, Benjamin J; Gouvas, Eftychia; Deliu, Zane; Garcia, Kelly D; Pestle, William J; Radosevich, James A

    2012-09-01

    Electrocautery and directed energy devices (DEDs) such as lasers, which are used in surgery, result in tissue damage that cannot be readily detected by traditional histological methods, such as hematoxylin and eosin staining. Alternative staining methods, including 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to stain live tissue, have been reported. Despite providing superior detection of damaged tissue relative to the hematoxylin and eosin (H&E) method, the MTT method possesses a number of drawbacks, most notably that it must be carried out on live tissue samples. Herein, we report the development of a novel staining method, "antigen destruction immunohistochemistry" (ADI), which can be carried out on paraffin-embedded tissue. The ADI method takes advantage of epitope loss to define the area of tissue damage and provides many of the benefits of live tissue MTT staining without the drawbacks inherent to that method. In addition, the authors provide data to support the use of antibodies directed at a number of gene products for use in animal tissue for which there are no species-specific antibodies commercially available, as well as an example of a species-specific direct antibody. Data are provided that support the use of this method in many tissue models, as well as evidence that ADI is comparable to the live tissue MTT method.

  4. Image processing in digital pathology: an opportunity to solve inter-batch variability of immunohistochemical staining

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    van Eycke, Yves-Rémi; Allard, Justine; Salmon, Isabelle; Debeir, Olivier; Decaestecker, Christine

    2017-02-01

    Immunohistochemistry (IHC) is a widely used technique in pathology to evidence protein expression in tissue samples. However, this staining technique is known for presenting inter-batch variations. Whole slide imaging in digital pathology offers a possibility to overcome this problem by means of image normalisation techniques. In the present paper we propose a methodology to objectively evaluate the need of image normalisation and to identify the best way to perform it. This methodology uses tissue microarray (TMA) materials and statistical analyses to evidence the possible variations occurring at colour and intensity levels as well as to evaluate the efficiency of image normalisation methods in correcting them. We applied our methodology to test different methods of image normalisation based on blind colour deconvolution that we adapted for IHC staining. These tests were carried out for different IHC experiments on different tissue types and targeting different proteins with different subcellular localisations. Our methodology enabled us to establish and to validate inter-batch normalization transforms which correct the non-relevant IHC staining variations. The normalised image series were then processed to extract coherent quantitative features characterising the IHC staining patterns.

  5. Image processing in digital pathology: an opportunity to solve inter-batch variability of immunohistochemical staining.

    Science.gov (United States)

    Van Eycke, Yves-Rémi; Allard, Justine; Salmon, Isabelle; Debeir, Olivier; Decaestecker, Christine

    2017-02-21

    Immunohistochemistry (IHC) is a widely used technique in pathology to evidence protein expression in tissue samples. However, this staining technique is known for presenting inter-batch variations. Whole slide imaging in digital pathology offers a possibility to overcome this problem by means of image normalisation techniques. In the present paper we propose a methodology to objectively evaluate the need of image normalisation and to identify the best way to perform it. This methodology uses tissue microarray (TMA) materials and statistical analyses to evidence the possible variations occurring at colour and intensity levels as well as to evaluate the efficiency of image normalisation methods in correcting them. We applied our methodology to test different methods of image normalisation based on blind colour deconvolution that we adapted for IHC staining. These tests were carried out for different IHC experiments on different tissue types and targeting different proteins with different subcellular localisations. Our methodology enabled us to establish and to validate inter-batch normalization transforms which correct the non-relevant IHC staining variations. The normalised image series were then processed to extract coherent quantitative features characterising the IHC staining patterns.

  6. Image processing in digital pathology: an opportunity to solve inter-batch variability of immunohistochemical staining

    Science.gov (United States)

    Van Eycke, Yves-Rémi; Allard, Justine; Salmon, Isabelle; Debeir, Olivier; Decaestecker, Christine

    2017-01-01

    Immunohistochemistry (IHC) is a widely used technique in pathology to evidence protein expression in tissue samples. However, this staining technique is known for presenting inter-batch variations. Whole slide imaging in digital pathology offers a possibility to overcome this problem by means of image normalisation techniques. In the present paper we propose a methodology to objectively evaluate the need of image normalisation and to identify the best way to perform it. This methodology uses tissue microarray (TMA) materials and statistical analyses to evidence the possible variations occurring at colour and intensity levels as well as to evaluate the efficiency of image normalisation methods in correcting them. We applied our methodology to test different methods of image normalisation based on blind colour deconvolution that we adapted for IHC staining. These tests were carried out for different IHC experiments on different tissue types and targeting different proteins with different subcellular localisations. Our methodology enabled us to establish and to validate inter-batch normalization transforms which correct the non-relevant IHC staining variations. The normalised image series were then processed to extract coherent quantitative features characterising the IHC staining patterns. PMID:28220842

  7. Basement membrane changes in breast cancer detected by immunohistochemical staining for laminin

    DEFF Research Database (Denmark)

    Albrechtsen, R; Nielsen, M; Wewer, U

    1981-01-01

    The distribution of the basement membrane glycoprotein laminin was studied by the immunoperoxidase technique in benign and malignant human breast tissue and in axillary lymph nodes from patients with breast cancer. An antiserum prepared against rat laminin was used. The specificity...... of this antiserum against human laminin was studied using the FL cell line of human epithelial-like cells derived from normal amniotic membrane. The antiserum reacted with these cells in immunoperoxidase staining and precipitated metabolically labeled secreted polypeptides which comigrated with polypeptides...... with molecular weights of 400,000 and 200,000 of rat laminin in sodium dodecyl sulfate:polyacrylamide gel electrophoresis. The neoplastic cells in malignant breast tissues showed strong cytoplasmic staining for laminin, and a positive reaction was aslo found in lymph node metastases. In some cases in which only...

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

    Science.gov (United States)

    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.

  9. Datamining approach for automation of diagnosis of breast cancer in immunohistochemically stained tissue microarray images.

    Science.gov (United States)

    Prasad, Keerthana; Zimmermann, Bernhard; Prabhu, Gopalakrishna; Pai, Muktha

    2010-05-28

    Cancer of the breast is the second most common human neoplasm, accounting for approximately one quarter of all cancers in females after cervical carcinoma. Estrogen receptor (ER), Progesteron receptor and human epidermal growth factor receptor (HER-2/neu) expressions play an important role in diagnosis and prognosis of breast carcinoma. Tissue microarray (TMA) technique is a high throughput technique which provides a standardized set of images which are uniformly stained, facilitating effective automation of the evaluation of the specimen images. TMA technique is widely used to evaluate hormone expression for diagnosis of breast cancer. If one considers the time taken for each of the steps in the tissue microarray process workflow, it can be observed that the maximum amount of time is taken by the analysis step. Hence, automated analysis will significantly reduce the overall time required to complete the study. Many tools are available for automated digital acquisition of images of the spots from the microarray slide. Each of these images needs to be evaluated by a pathologist to assign a score based on the staining intensity to represent the hormone expression, to classify them into negative or positive cases. Our work aims to develop a system for automated evaluation of sets of images generated through tissue microarray technique, representing the ER expression images and HER-2/neu expression images. Our study is based on the Tissue Microarray Database portal of Stanford university at http://tma.stanford.edu/cgi-bin/cx?n=her1, which has made huge number of images available to researchers. We used 171 images corresponding to ER expression and 214 images corresponding to HER-2/neu expression of breast carcinoma. Out of the 171 images corresponding to ER expression, 104 were negative and 67 were representing positive cases. Out of the 214 images corresponding to HER-2/neu expression, 112 were negative and 102 were representing positive cases. Our method has 92

  10. Diagnostic value of SOX-10 immunohistochemical staining for the detection of uveal melanoma.

    Science.gov (United States)

    Alghamdi, Sarah A; Zoroquiain, Pablo; Dias, Ana Beatriz T; Alhumaid, Sulaiman R; Aldrees, Sultan; Burnier, Miguel N

    2015-01-01

    SOX-10 has been shown to be a sensitive marker of cutaneous melanoma. This study aimed to evaluate Sox-10 expression in uveal melanoma. A total of 40 tissue blocks of enucleated eyes with uveal melanoma were cut and stained using an anti-SOX-10 mouse monoclonal antibody and HMB-45 antibody. SOX-10 showed exclusive nuclear positivity in 100% of the uveal melanoma cases (38/38). HMB-45 showed cytoplasmic positivity in 97.3 (37/38). Positivity for SOX-10 was also noted in the inner and outer nuclear layers of the retina in 78% of the enucleated eyes. SOX-10 expression proved to be the most sensitive marker for uveal melanoma, and therefore, we propose a modified panel for the diagnosis of uveal melanoma that includes both SOX-10 and HMB-45. The observation of distinct, diffuse nuclear SOX-10 expression in retinal inner and outer nuclear layers is a finding that warrants further investigation as a marker for retinoblastoma.

  11. Use of immunohistochemical staining and electron microscopy to aid in diagnosis of soft tissue sarcomas associated with the fetlock joint in two horses.

    Science.gov (United States)

    Findley, Judith A; Singer, Ellen R; Milner, Peter I; Leeming, Gail H

    2014-05-01

    Soft tissue sarcomas of the equine distal limb associated with joints, sheaths, or bursae have rarely been reported. Accurate diagnosis of these tumors is challenging in both human beings and veterinary species. Immunohistochemical staining and transmission electron microscopy have been used in human beings to reduce misdiagnosis. The current report describes 2 mature horses presenting with lameness and swelling associated with the dorsal aspect of the metacarpo(tarso)phalangeal joint. In both cases, surgical excision was performed with subsequent histological analysis of the masses to determine the tissue of origin. In both cases, immunohistochemical staining and transmission electron microscopy aided the definitive diagnosis of fibrosarcoma associated with the fetlock joints of 2 horses. © 2014 The Author(s).

  12. A list of image files of planarians analyzed by in situ hybridication and immunohistochemical staining - Plabrain DB | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available switchLanguage; BLAST Search Image Search Home About Archive Update History Data List Contact us Plab...ata file File name: planaria_image_list.zip File URL: ftp://ftp.biosciencedbc.jp/archive/plabrain-db/LATEST/...planaria_image_list.zip File size: 1KB Simple search URL http://togodb.biosciencedbc.jp/togodb/view/plabrain...te Policy | Contact Us A list of image files of planarians analyzed by in situ hybridication and immunohistochemical staining - Plabrain DB | LSDB Archive ...

  13. Image files of planarians analyzed by in situ hybridication and immunohistochemical staining - Plabrain DB | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available switchLanguage; BLAST Search Image Search Home About Archive Update History Data List Contact us Plab...]regeneration[days after decapitation ].jpg Data file File name: planaria_image.zip File URL: ftp://ftp.biosciencedbc.jp/archive/plab...ate History of This Database Site Policy | Contact Us Image files of planarians analyzed by in situ hybridication and immunohistochemical staining - Plabrain DB | LSDB Archive ...

  14. Method for the validation of immunohistochemical staining using SCID mouse xenografts: expression of CD40 and CD154 in human non-small cell lung cancer.

    Science.gov (United States)

    Ishikawa, Keidai; Miyamoto, Masaki; Yoshioka, Tatsuya; Kadoya, Masatoshi; Li, Li; Mishra, Roshan; Ichinokawa, Kazuomi; Shoji, Yasuhito; Matsumura, Yoshiyuki; Hida, Yasuhiro; Kaga, Kichizo; Kato, Tatsuya; Kaji, Mitsuhito; Ohbuchi, Toshiro; Itoh, Tomoo; Dosaka-Akita, Hirotoshi; Matsui, Yoshiro; Hirano, Satoshi

    2013-04-01

    This report proposes a concept for the standardization of immunohistochemical evaluation. Immunohistochemical staining has several problems associated with the sensitivity of the technical process and standardization of the assessment of potent staining. We provided data focusing on this concept through immunostaining for CD154 in non-small cell lung cancer (NSCLC). We used two types of anti-CD154 antibody as primary antibodies in immunohistochemical staining, as previously reported. Western blot analysis confirmed strong CD154 expression in the cultured cell line PC10, but not in LK2. We also assessed CD154 expression in SCID mouse xenografts of these cell lines. SCID xenograft data on western blot analysis were consistent with those of cultured cell lines. These xenografts could thus be used as positive or negative tissue controls for CD154 immunostaining. Primary antibodies should therefore be confirmed as recognizing target lesions, while control tissue specimens should be objectively confirmed as having target products using another experimental method. Our method would allow results to be unified at more than one laboratory and could act as an objective control assessment method in immunohistochemistry.

  15. Antibody validation and scoring guidelines for ABCG2 immunohistochemical staining in formalin-fixed paraffin-embedded colon cancer tissue

    DEFF Research Database (Denmark)

    Cederbye, Camilla Natasha; Palshof, Jesper Andreas; Hansen, Tine Plato

    2016-01-01

    Overexpression of the ATP-dependent drug efflux pump ABCG2 is a major molecular mechanism of multidrug resistance in cancer and might be a predictive biomarker for drug response. Contradictory results have been reported for immunohistochemical studies of ABCG2 protein expression in colorectal can...

  16. QUANTIFICATION OF AREA PERCENTAGE OF IMMUNOHISTOCHEMICAL STAINING BY TRUE COLOR IMAGE-ANALYSIS WITH APPLICATION OF FIXED THRESHOLDS

    NARCIS (Netherlands)

    WILLEMSE, F; HENZENLOGMANS, SC; EGGINK, HF

    1994-01-01

    Most image analysis systems (IAS) use black-and-white cameras. However, true color IASs are considered to be useful for quantification of immunohistologically stained structures. Using a true color IAS, we evaluated two methods of segmentation for quantification of area percentage of staining: one

  17. Comparative evaluation of eosinophils in normal mucosa, dysplastic mucosa and oral squamous cell carcinoma with hematoxylin-eosin, Congo red, and EMR1 immunohistochemical staining techniques.

    Science.gov (United States)

    Kargahi, Neda; Razavi, Sayyed Mohammad; Deyhimi, Parviz; Homayouni, Solmaz

    2015-01-01

    Oral squamous cell carcinoma is the most common malignant lesion of the oral cavity, and it involves various molecular mechanisms. The development of oral squamous cell carcinoma is influenced by the host immune cells, such as eosinophils. The present study was conducted to compare the presence of eosinophils in normal mucosa, dysplastic mucosa, and oral squamous cell carcinoma by -hematoxylin- eosin staining, Congo red staining, and epidermal growth factor-like (EGF-like) module containing a mucin-like hormone receptor1 (EMR1) immunohistochemical marker. In this cross-sectional study, 60 paraffinized samples were selected, consisting of 20 normal mucosae, 20 dysplastic mucosae, and 20 squamous cell carcinoma samples. After confirmation of the diagnosis, the mean number of eosinophils was evaluated by hematoxylin-eosin, Congo red, and immunohystochemical staining techniques. The data were analyzed by SPSS-10 software using the Kruskal-Wallis and Friedman tests. The results showed that the number of eosinophils in dysplastic mucosa was significantly higher than the number in normal mucosa, and the number of eosinophils in squamous cell carcinoma was significantly higher than the number in dysplastic mucosa in all staining techniques (poral lesions in cases with borderline diagnosis and in targeted molecular therapy.

  18. Antigen retrieval in formalin-fixed, paraffin-embedded tissues: an enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections.

    Science.gov (United States)

    Shi, S R; Key, M E; Kalra, K L

    1991-06-01

    We describe a new approach for retrieval of antigens from formalin-fixed, paraffin-embedded tissues and their subsequent staining by immunohistochemical techniques. This method of antigen retrieval is based on microwave heating of tissue sections attached to microscope slides to temperatures up to 100 degrees C in the presence of metal solutions. Among 52 monoclonal and polyclonal antibodies tested by this method, 39 antibodies demonstrated a significant increase in immunostaining, nine antibodies showed no change, and four antibodies showed reduced immunostaining. In particular, excellent immunostaining results were obtained with a monoclonal antibody to vimentin as well as several different keratin antibodies on routine formalin-fixed tissue sections after pre-treatment of the slides with this method. These results showed that after antigen retrieval: (a) enzyme predigestion of tissues could be omitted; (b) incubation times of primary antibodies could be significantly reduced, or dilutions of primary antibodies could be increased; (c) adequate staining could be achieved in long-term formalin-fixed tissues that failed to stain by conventional methods; and (d) certain antibodies which were typically unreactive with formalin-fixed tissues gave excellent staining.

  19. TP53 Staining in Tissue Samples of Chronic Lymphocytic Lymphoma Cases: An Immunohistochemical Survey of 51 Cases

    Directory of Open Access Journals (Sweden)

    İbrahim Kulaç

    2017-03-01

    Full Text Available Objective: Chronic lymphocytic leukemia (CLL is the most common lymphoproliferative disease in adults. The aim of this study is to find out if the extent of proliferation centers or the immunohistochemical expression of p53 is related to disease prognosis. Materials and Methods: In the scope of this study, 54 biopsy specimens from 51 patients (50 of lymph nodes; the others of spleen, tonsil, orbit, and liver diagnosed with CLL at the Hacettepe University Department of Pathology in 2000-2013 were reevaluated. The clinical and demographic data of the patients were obtained from our patient database. Biopsy samples were assessed semi-quantitatively for the percentage of proliferation center/total biopsy area (PC/TBA and an immunohistochemical study was performed on representative blocks of tissues for p53 expression. Results: When the patients were divided into two categories according to Rai stage as high and low (stages 0, 1, and 2 vs. stages 3 and 4, it was seen that patients with low Rai stage had a better prognosis than those with high stages (p=0.030. However, there was no statistically significant correlation between overall survival and PC/TBA ratio or p53 expression levels. Conclusion: In our cohort, PC/TBA ratio and immunopositivity of p53 did not show correlations with overall survival.

  20. Immunohistochemical staining for thyroid peroxidase (TPO) of needle core biopsies in the diagnosis of scintigraphically cold thyroid nodules

    DEFF Research Database (Denmark)

    Yousaf, U.; Christensen, Lars Høj; Rasmussen, A.K.

    2008-01-01

    BACKGROUND: Cold thyroid nodules are common, in particular in iodine-deficient areas, but only a minority of them are malignant requiring surgery. Thyroid peroxidase (TPO) immunostaining of fine-needle aspiration cytology (FNAC) material has proven helpful in diagnosing cells from malignant lesions......, but the procedure has its limitations in a routine setting. PURPOSE: To improve diagnosis and reduce surgery rate, the FNAC procedure was replaced by needle core biopsy (NCB), which was routinely stained for TPO by the monoclonal antibody mAb 47. MATERIALS AND METHODS: During a 5-year period 427 consecutive...... patients with a cold thyroid nodule were evaluated by ultrasound-guided NCB, which had been routinely stained for TPO in an automated immunostainer. Sensitivity and specificity and predictive values of the TPO immunostaining were estimated, based on the final diagnosis obtained from surgical resection...

  1. Immunohistochemical staining of leptin is associated with grade, stage, lymph node involvement, recurrence, and hormone receptor phenotypes in breast cancer

    OpenAIRE

    Khabaz, Mohamad Nidal; Abdelrahman, Amer; Butt, Nadeem; Damnhory, Lila; Elshal, Mohamed; Aldahlawi, Alia M.; Ashoor, Swsan; Al-Maghrabi, Basim; Dobson, Pauline; Brown, Barry; Al-Sakkaf, Kaltoom; Al-Qahtani, Mohmmad; Al-Maghrabi, Jaudah

    2017-01-01

    Background Obesity is part of the established risk factors for breast cancer (BC) in postmenopausal females. Circulating leptin increases in parallel with the increase of body weight and fat reservoir. Methods This research investigated the link between leptin phenotype and the clinicopathological factors in BC. A large set of breast cancer cases (449), and 27 non-cancerous tissue samples of breast were employed for leptin expression recognition using immunohistochemistry staining. Results Cy...

  2. Correlation between immunohistochemical staining of CEACAM1 and clinicopathological findings in oral pre-neoplastic lesions and squamous cell carcinoma.

    Science.gov (United States)

    Simonetti, Oriana; Lucarini, Guendalina; Rubini, Corrado; Zizzi, Antonio; Aspriello, Simone Domenico; Di Primio, Roberto; Offidani, Anna Maria

    2017-09-08

    Squamous cell carcinoma of the oral cavity represents the sixth most common cancer worldwide and it is often preceded by pre-neoplastic lesions. Sometimes it is still difficult for pathologists to make objective differential diagnoses only on histological characteristics. Tumorigenesis is accompanied by altered expression of cell adhesion molecules, like carcinoembryonic antigen cell adhesion molecule (CEACAM)1. We wanted to investigative CEACAM1 in oral dysplastic lesions, carcinoma in situ (CIS) and oral squamous cell carcinoma (OSCC). We examined immunohistochemical CEACAM1 expression in 50 OSCC, 30 oral CIS and 40 pre-neoplastic lesions and assessed its correlation with clinical and pathological parameters. CEACAM1 was not expressed in normal mucosa, significantly expressed in CIS while it was negative in all the dysplastic lesions. In OSCC, high CEACAM1 expression was associated with tumor grade and inversely correlated with both overall and disease-specific 5-year survival. We showed that CEACAM1 expression is very dynamic: absent in dysplastic lesions, up-regulated in CIS and OSCC. We suggest that CEACAM1 could be a prognostic marker of OSCC and oral CIS. Our most important finding was that it could help pathologists diagnosing oral carcinoma in situ.

  3. Immunohistochemical staining for thyroid peroxidase (TPO) of needle core biopsies in the diagnosis of scintigraphically cold thyroid nodules.

    Science.gov (United States)

    Yousaf, U; Christensen, L H; Rasmussen, A K; Jensen, F; Mollerup, C L; Kirkegaard, J; Lausen, I; Rank, F; Feldt-Rasmussen, U

    2008-06-01

    Cold thyroid nodules are common, in particular in iodine-deficient areas, but only a minority of them are malignant requiring surgery. Thyroid peroxidase (TPO) immunostaining of fine-needle aspiration cytology (FNAC) material has proven helpful in diagnosing cells from malignant lesions, but the procedure has its limitations in a routine setting. To improve diagnosis and reduce surgery rate, the FNAC procedure was replaced by needle core biopsy (NCB), which was routinely stained for TPO by the monoclonal antibody mAb 47. During a 5-year period 427 consecutive patients with a cold thyroid nodule were evaluated by ultrasound-guided NCB, which had been routinely stained for TPO in an automated immunostainer. Sensitivity and specificity and predictive values of the TPO immunostaining were estimated, based on the final diagnosis obtained from surgical resection. The majority of nodules with benign NCB diagnosis were not surgically removed, and thus a subgroup of 140 operated nodules formed the basis for the calculations. Sensitivity and specificity for benign and malignant lesions were 100% if the oxyphilic variant of adenomas and minimally invasive follicular carcinomas were excluded. By inclusion of these, the values fell to 89% and 97%, respectively. The predictive value of a positive test was 96% and the predictive value of a negative test was 97%. TPO immunostaining was found to be a valuable adjunct to morphology in the diagnosis of cold thyroid nodules of the nonoxyphilic type.

  4. Study of Estrogen Receptor and Progesterone Receptor Expression in Breast Ductal Carcinoma In Situ by Immunohistochemical Staining in ER/PgR-Negative Invasive Breast Cancer.

    Science.gov (United States)

    Dobrescu, Andrei; Chang, Monique; Kirtani, Vatsala; Turi, George K; Hennawy, Randa; Hindenburg, Alexander A

    2011-01-01

    Background. To our knowledge, the hormone receptor status of noncontiguous ductal carcinoma in situ (DCIS) occurring concurrently in ER/PgR-negative invasive cancer has not been studied. The current study was undertaken to investigate the ER/PgR receptor status of DCIS of the breast in patients with ER/PgR-negative invasive breast cancer. Methods. We reviewed the immunohistochemical (IHC) staining for ER and PgR of 187 consecutive cases of ER/PgR-negative invasive breast cancers, collected from 1995 to 2002. To meet the criteria for the study, we evaluated ER/PgR expression of DCIS cancer outside of the invasive breast cancer. Results. A total of 37 cases of DCIS meeting the above criteria were identified. Of these, 16 cases (43.2%) showed positive staining for ER, PgR, or both. Conclusions. In our study of ER/PgR-negative invasive breast cancer we found that in 8% of cases noncontiguous ER/PR-positive DCIS was present. In light of this finding, it may be important for pathologists to evaluate the ER/PgR status of DCIS occurring in the presence of ER/PgR-negative invasive cancer, as this subgroup could be considered for chemoprevention.

  5. Comparison of photoacoustically derived hemoglobin and oxygenation measurements with contrast-enhanced ultrasound estimated vascularity and immunohistochemical staining in a breast cancer model.

    Science.gov (United States)

    Eisenbrey, John R; Merton, Daniel A; Marshall, Andrew; Liu, Ji-Bin; Fox, Traci B; Sridharan, Anush; Forsberg, Flemming

    2015-01-01

    In this preliminary study, we compared two noninvasive techniques for imaging intratumoral physiological conditions to immunohistochemical staining in a murine breast cancer model. MDA-MB-231 tumors were implanted in the mammary pad of 11 nude rats. Ultrasound and photoacoustic (PA) scanning were performed using a Vevo 2100 scanner (Visualsonics, Toronto, Canada). Contrast-enhanced ultrasound (CEUS) was used to create maximum intensity projections as a measure of tumor vascularity. PAs were used to determine total hemoglobin signal (HbT), oxygenation levels in detected blood (SO2 Avg), and oxygenation levels over the entire tumor area (SO2 Tot). Tumors were then stained for vascular endothelial growth factor (VEGF), cyclooxygenase-2 (Cox-2), and the platelet endothelial cell adhesion molecule CD31. Correlations between findings were analyzed using Pearson's coefficient. Significant correlation was observed between CEUS-derived vascularity measurements and both PA indicators of blood volume (r = 0.49 for HbT, r = 0.50 for SO2 Tot). Cox-2 showed significant negative correlation with SO2 Avg (r = -0.49, p = 0.020) and SO2 Tot (r = -0.43, p = 0.047), while CD31 showed significant negative correlation with CEUS-derived vascularity (r = -0.47, p = 0.036). However, no significant correlation was observed between VEGF expression and any imaging modality (p > 0.08). Photoacoustically derived HbT and SO2 Tot may be a good indicator of tumor fractional vascularity. While CEUS correlates with CD31 expression, photoacoustically derived SO2 Avg appears to be a better predictor of Cox-2 expression. © The Author(s) 2014.

  6. The utility of PAX8 and IMP3 immunohistochemical stains in the differential diagnosis of benign, premalignant, and malignant endocervical glandular lesions.

    Science.gov (United States)

    Danialan, Richard; Assaad, Margaret; Burghardt, Jason; Newcomb, Pamela; Cartun, Richard W; Mandavilli, Srinivas

    2013-08-01

    Glandular lesions of the endocervix can be diagnostically challenging and occasionally the differential diagnosis includes endocervical adenocarcinoma in situ (EC AIS) and well-differentiated endocervical adenocarcinoma (ECA). PAX8 and IMP3 are two markers which have not been well studied in the endocervix. Our aim was to evaluate their immunohistochemical (IHC) expression in benign and malignant endocervical glandular lesions as well as to compare them to the traditionally used panel (Ki-67, p16, CEA). We searched our surgical pathology files for a cohort of benign endocervical glandular lesions as well as premalignant and malignant groups including EC AIS and ECA. An IHC panel consisting of PAX8, IMP3, Ki-67, p16, and CEA was performed on all cases. Immunoreactivity was scored on a degree of positivity (S0=no immunoreactivity, S1=up to 10% cells, S2=between 10 and 50% cells, S3=>50% cells) and intensity (Int0 - absent, Int1 - mild/faint, Int2 - moderate, Int3 - strong). PAX8 showed diffuse positivity (S3) with at least a moderate intensity of staining (Int2) in the benign group. PAX8 was focal (S1) in ECA and faint (Int1), compared to EC AIS, which was moderate (S2) and faint (Int1). IMP3 expression was focal in the benign group (S1), moderate (S2) in EC AIS and moderate-to-diffuse (S2-3) in ECA. IMP3 intensity was faint (Int1) in benign lesions, moderate (Int2) in EC AIS, and strong (Int3) in ECA. Significant Ki-67, p16, and CEA expression was noted in the premalignant/malignant cohort. PAX8 and IMP3 can be helpful in the differential diagnosis of benign vs. malignant endocervical glandular lesions. Our study, however, shows that there is some degree of overlap of staining in both the benign and malignant group. As such, PAX8 and IMP3 should always be interpreted with caution and in combination with the histomorphology. Published by Elsevier Inc.

  7. Immunohistochemical Ki-67/KL1 double stains increase accuracy of Ki-67 indices in breast cancer and simplify automated image analysis

    DEFF Research Database (Denmark)

    Nielsen, Patricia S; Bentzer, Nina K; Jensen, Vibeke

    2014-01-01

    by digital image analysis. This study aims to detect the difference in accuracy and precision between manual indices of single and double stains, to develop an automated quantification of double stains, and to explore the relation between automated indices and tumor characteristics when quantified...... observers and automated image analysis. RESULTS: Indices were predominantly higher for single stains than double stains (P≤0.002), yet the difference between observers was statistically significant (Pcorrelation coefficient for manual and automated indices ranged from 0.......69 to 0.85 (Pcorrelating automated indices with tumor characteristics, for example, tumor size (P

  8. Immunohistochemical study of Ulex europaeus agglutinin 1 (UEA-1) binding of megakaryocytes in bone marrow biopsy specimens: demonstration of heterogeneity in staining pattern reflecting the stages of differentiation.

    Science.gov (United States)

    Liu, S M; Li, C Y

    1996-01-01

    During differentiation, megakaryocytes undergo nuclear endoreplication, an increase in cell size, cytoplasmic granulation, and release of platelets. The changes in highly lobulated nuclei with varying degree of polyploidy and increasing cell size are easily recognized morphologically. However, the actual cytoplasmic changes are more difficult to perceive morphologically. With the peroxidase-antiperoxidase (PAP) method using UEA-1 as the binding protein to the alpha-L-fucose of glycoprotein synthesized by megakaryocytes, we observed significant variation in cytoplasmic staining of megakaryocytes in routinely processed bone marrow biopsy sections. A total of 3344 megakaryocytes in bone marrow sections from 10 patients with nonhematologic diseases and from 10 patients with idiopathic thrombocytopenic purpura (ITP) was studied. According to the intensity and pattern of cytoplasmic staining, we divided megakaryocytes into at least six groups: (1) low granular (LG), (2) diffuse granular (DG), (3) diffuse dense granular (DDG), (4) marginal granular (MG), (5) denuded (DMK), and (6) endomitotic (EndoM). Most of the megakaryocytes were DG (mean, 42.75% +/- 19.21%) and DDG (mean, 50.25% +/- 21.23%). In correlation with nuclear morphology and cell size, it appears that substances binding to UEA-1 are located in the paranuclear region in early megakaryocytes and produce a low granular focal staining pattern (LG cells). Next, the granules spread throughout the cytoplasm (DG cells) and increase in quantity (DDG). This is followed by migration of granules to the periphery of the cytoplasm (MG cells) and is associated with the liberation of platelets and eventual formation of DMK megakaryocytes. Endomitosis, regulated by unknown factors, occurred in the MG stage. In comparing the group with nonhematologic disease (mean DG, 35.4% +/- 18.48%; DDG, 58.4% +/- 21.8%) and the group with ITP (mean DG, 50.1% +/- 17.82%; DDG, 42.1% +/- 18.12%), we found an increasing proportion of DG

  9. Objective measurement of the different collagen types in the corpus cavernosum of potent and impotent men: an immunohistochemical staining with computerized-image analysis.

    Science.gov (United States)

    Raviv, G; Kiss, R; Vanegas, J P; Petein, M; Danguy, A; Schulman, C; Wespes, E

    1997-01-01

    Quantitative measurements of the collagen types (I, II, and IV) in the corpora cavernosa of potent and impotent men were carried out to investigate whether quantitative immunohistochemistry might contribute additional information as to the cause of erectile dysfunction. The study group consisted of 22 men with various etiologies of impotence and 4 normal, potent men. The quantitative immunohistochemistry measurements were performed by means of a cell-image processor. Three variables for each of the three types of collagen were studied, namely, the mean optical density (MOD), which relates to histochemical staining intensity; the labeling index (LI), which is positively related to the percentage of immunostaining; and the quick score (QS) index, which takes into account both LI and MOD values. None of the quantitative parameters taken individually (monovariate statistical analyses) made it possible to obtain any statistically significant difference between the types of collagen of the group under study. The mean QS value recorded for collagen type IV was significantly lower than that noted for collagen type I in the psychogenic (P = 0.019), arteriogenic (P = 0.012), and venogenic (P = 0.001) groups, whereas the MOD value was significantly lower in the normal (P = 0.043), arteriogenic (P = 0.013), and venogenic (P = 0.001) groups but not in the psycogenic group. The mean MOD of collagen type III was intermediate between that of the other types. In contrast, the mean LI value recorded for collagen type IV was significantly lower only in the venogenic (P = 0.032) and psychogenic (P = 0.049) groups as compared with the other groups. No objective qualitative change in the collagen types was observed that could be correlated to the etiology of erectile dysfunction. The significant difference seen in the quantitative parameters with regard to collagen type IV and the observed increase in the type I/III collagen ratio might attest to the notion that the response of the

  10. Variations in immunohistochemical preservation of proteins in a mummification model

    National Research Council Canada - National Science Library

    Metcalfe, Ryan; Freemont, Tony

    2012-01-01

    ... and thereby improve confidence that the results are genuine. A mouse model of Egyptian mummification has been used to demonstrate that the survival of proteins, judged by the retention of immunohistochemical staining, varies markedly...

  11. Port-Wine Stains

    Science.gov (United States)

    ... for the Flu Vaccine? Eating Disorders Arrhythmias Port-Wine Stains KidsHealth > For Parents > Port-Wine Stains Print ... Manchas de vino de oporto What Are Port-Wine Stains? A port-wine stain is a type ...

  12. CD3 immunohistochemical staining in diagnosis of lymphocytic colitis

    DEFF Research Database (Denmark)

    Fiehn, Anne-Marie Kanstrup; Engel, Ulla; Holck, Susanne

    2016-01-01

    Microscopic colitis (MC) is a common cause of chronic watery diarrhea. Traditionally, MC encompasses the 2 subgroups lymphocytic colitis (LC) and collagenous colitis, but recently, an additional subgroup, MC incomplete, has been introduced. Distinguishing between the subgroups relies exclusively...

  13. Morphological and immunohistochemical analysis of apoptosis in the cerebellum of rats subjected to focal cerebral ischemia with or without alcoholism model.

    Science.gov (United States)

    Carvalho, Camila Albuquerque Melo de; Tirapelli, Daniela Pretti da Cunha; Rodrigues, Andressa Romualdo; Lizarte, Fermino Sanches; Novais, Paulo Cézar; Silva, Jairo Pinheiro; Carlotti, Carlos Gilberto; Colli, Benedicto Oscar; Tirapelli, Luís Fernando

    2016-09-01

    To evaluated histopathological changes, morphometric and expression of proteins CASPASE-3, BCL-2 and XIAP related to apoptosis in the cerebellum after induction of temporary focal cerebral ischemia followed by reperfusion, with or without a model of chronic alcoholism. Fifty Wistar rats were used and divided into: control group (C), sham group (S), ischemic group (I), alcoholic group (A), and ischemic and alcoholic group (IA). The cerebellum samples collected were stained for histopathological and morphometric analysis and immunohistochemistry study. Histopathological changes were observed a greater degree in animals in groups A and IA. The morphometric study showed no difference in the amount of cells in the granular layer of the cerebellum between the groups. The expression of CASPASE-3 was higher than BCL-2 and XIAP in the groups A and IA. We observed correlation between histopathological changes and the occurrence of apoptosis in cerebellar cortex.

  14. Pericardial fluid Gram stain

    Science.gov (United States)

    ... a smear. A series of special stains are applied to the sample. This is called a Gram stain . A laboratory specialist looks at the stained slide under the microscope, checking for bacteria. The color, size, and shape of the cells ...

  15. Automated quantification of nuclear immunohistochemical markers with different complexity.

    Science.gov (United States)

    López, Carlos; Lejeune, Marylène; Salvadó, María Teresa; Escrivà, Patricia; Bosch, Ramón; Pons, Lluis E; Alvaro, Tomás; Roig, Jordi; Cugat, Xavier; Baucells, Jordi; Jaén, Joaquín

    2008-03-01

    Manual quantification of immunohistochemically stained nuclear markers is still laborious and subjective and the use of computerized systems for digital image analysis have not yet resolved the problems of nuclear clustering. In this study, we designed a new automatic procedure for quantifying various immunohistochemical nuclear markers with variable clustering complexity. This procedure consisted of two combined macros. The first, developed with a commercial software, enabled the analysis of the digital images using color and morphological segmentation including a masking process. All information extracted with this first macro was automatically exported to an Excel datasheet, where a second macro composed of four different algorithms analyzed all the information and calculated the definitive number of positive nuclei for each image. One hundred and eighteen images with different levels of clustering complexity was analyzed and compared with the manual quantification obtained by a trained observer. Statistical analysis indicated a great reliability (intra-class correlation coefficient > 0.950) and no significant differences between the two methods. Bland-Altman plot and Kaplan-Meier curves indicated that the results of both methods were concordant around 90% of analyzed images. In conclusion, this new automated procedure is an objective, faster and reproducible method that has an excellent level of accuracy, even with digital images with a high complexity.

  16. TMARKER: A free software toolkit for histopathological cell counting and staining estimation

    Directory of Open Access Journals (Sweden)

    Peter J Schüffler

    2013-01-01

    Full Text Available Background: Histological tissue analysis often involves manual cell counting and staining estimation of cancerous cells. These assessments are extremely time consuming, highly subjective and prone to error, since immunohistochemically stained cancer tissues usually show high variability in cell sizes, morphological structures and staining quality. To facilitate reproducible analysis in clinical practice as well as for cancer research, objective computer assisted staining estimation is highly desirable. Methods: We employ machine learning algorithms as randomized decision trees and support vector machines for nucleus detection and classification. Superpixels as segmentation over the tissue image are classified into foreground and background and thereafter into malignant and benign, learning from the user′s feedback. As a fast alternative without nucleus classification, the existing color deconvolution method is incorporated. Results: Our program TMARKER connects already available workflows for computational pathology and immunohistochemical tissue rating with modern active learning algorithms from machine learning and computer vision. On a test dataset of human renal clear cell carcinoma and prostate carcinoma, the performance of the used algorithms is equivalent to two independent pathologists for nucleus detection and classification. Conclusion: We present a novel, free and operating system independent software package for computational cell counting and staining estimation, supporting IHC stained tissue analysis in clinic and for research. Proprietary toolboxes for similar tasks are expensive, bound to specific commercial hardware (e.g. a microscope and mostly not quantitatively validated in terms of performance and reproducibility. We are confident that the presented software package will proof valuable for the scientific community and we anticipate a broader application domain due to the possibility to interactively learn models for new

  17. Evaluation of alpha-synuclein immunohistochemical methods for the detection of Lewy-type synucleinopathy in gastrointestinal biopsies.

    Science.gov (United States)

    Corbillé, Anne-Gaëlle; Letournel, Franck; Kordower, Jeffrey H; Lee, John; Shanes, Elisheva; Neunlist, Michel; Munoz, David G; Derkinderen, Pascal; Beach, Thomas G

    2016-04-04

    The observation showing that Lewy type synucleinopathy (LTS), the pathological hallmark of Parkinson's disease (PD), is found in the gut of almost all PD subjects led to a substantial amount of research to develop a diagnostic procedure in living patients based on endoscopically obtained gastrointestinal biopsies. However, the existing studies have provided conflicting results regarding the sensitivity and specificity of gastrointestinal biopsies for the detection of LTS. We therefore undertook a multi-center staining and blinded judging of a common set of slides from colonic biopsies to determine the optimal protocol for the detection of LTS. Four different immunohistochemical methods, developed in four separate expert laboratories, were evaluated for their sensitivity and specificity to detect enteric LTS. Test sets of formalin-fixed, paraffin-embedded sections from biopsies of 9 PD subjects and 3 controls were stained with the 4 methods and graded by 4 different observers. Four types of staining morphology (granular staining in the lamina propria, perivascular/vascular wall staining in the submucosa, lacy-granular pattern in the submucosa and epithelial cell nuclear staining) were variably observed in the slides stained by the 4 methods. Positive alpha-synuclein staining was observed by all 5 judges in most of the slides from control cases, regardless of the staining methods that were used. Moreover, none of the tested method or staining pattern had a specificity and sensitivity more than 80 % regarding to PD. Overall, our study suggest that the tested methods are not adequate for the prediction of PD using gastrointestinal biopsies. Future studies are warranted to test new immunostaining methods.

  18. Subjectivity

    Directory of Open Access Journals (Sweden)

    Jesús Vega Encabo

    2015-11-01

    Full Text Available In this paper, I claim that subjectivity is a way of being that is constituted through a set of practices in which the self is subject to the dangers of fictionalizing and plotting her life and self-image. I examine some ways of becoming subject through narratives and through theatrical performance before others. Through these practices, a real and active subjectivity is revealed, capable of self-knowledge and self-transformation. 

  19. Combined beta-galactosidase and immunogold/silver staining for immunohistochemistry and DNA in situ hybridization

    NARCIS (Netherlands)

    van den Brink, W.; van der Loos, C.; Volkers, H.; Lauwen, R.; van den Berg, F.; Houthoff, H. J.; Das, P. K.

    1990-01-01

    A combination of beta-galactosidase enzyme and the immunogold/silver staining method was studied for evaluation of double-staining experiments. Applications are shown for immunohistochemical double staining using two monoclonal antibodies and for combined immunohistochemistry and DNA in situ

  20. Immunohistochemical detection of P53 and Mdm2 in vitiligo

    Directory of Open Access Journals (Sweden)

    Ola A Bakry

    2012-01-01

    Full Text Available Background: Vitiligo is a common depigmented skin disorder that is caused by selective destruction of melanocytes. It is generally accepted that the main function of melanin resides in the protection of skin cells against the deleterious effect of ultraviolet rays (UVRs. Association of vitiligo and skin cancer has been a subject of controversy. Occurrence of skin cancer in long-lasting vitiligo is rare despite multiple evidences of DNA damage in vitiliginous skin. Aim: To detect the expression of P53 and Mdm2 proteins in both depigmented and normally pigmented skin of vitiligo patients and to compare it to control subjects suffering from nonmelanoma skin cancer (NMSC. Materials and Methods: Thirty-four patients with vitiligo and 30 age and sex-matched patients with nodulo-ulcerative basal cell carcinoma (BCC as a control group were selected. Both patients and control subjects had outdoor occupations. Skin biopsies were taken from each case and control subjects. Histopathological examination of Hematoxylin and eosin-stained sections was done. Expression of P53 and Mdm2 proteins were examined immunohistochemically. Results: Both P53 and Mdm2 were strongly expressed in depigmented as well as normally pigmented skin of vitiligo patients. This expression involved the epidermis, skin adnexa and blood vessels with significant differences between cases and controls. Conclusions: The overexpression of P53 and Mdm2 proteins in both normally pigmented and depigmented skin of patients with vitiligo could contribute to the decreased occurrence of actinic damage and NMSC in these patients.

  1. IMMUNOHISTOCHEMICAL ANALYSIS OF ORAL MUCOSA LEUKOPLAKIA

    Directory of Open Access Journals (Sweden)

    Yu. G. KOLENKO

    2016-06-01

    Full Text Available In recent years, substantial changes have occurred in the structure of oral mucosa diseases, in particular an increased ratio of precancerous diseases, so that an effective non-invasive detection of any sign of malignancy appears as an urgent and most actual task of dentistry. Aim: To study the proliferative activity of epithelial cells in Ki-67 antigenin patients with leukoplakia of the oral mucosa. Materials and method: A complex clinical and laboratory examination was performed on 155 patients with oral leukoplakia, who addressed the Operative Dentistry Department of the “A.A.Bogomolets” National Medical University of Kiev between 2010 and 2014. All patients have been subjected to a careful clinical examination, which included: dental anamnesis, visual inspection, oral examination and digital palpation of oral mucosa and tongue mucosa, biopsy of leukoplakia lesions for cytological and histological examination. Results: Histological evaluation of the material has been performed according to the WHO (2005 classification of leukoplakia. 10 (14% sites of unaltered mucosa, 10 (14% samples of hyperkeratosis without atypia, 14 (19% biopsy specimens of hyperkeratosis SIN1, 15 (21% – hyperkeratosis SIN2, 10 (14% - SIN3 and 13 (18% cases of squamous cell carcinoma were evidenced. Immunohistochemical investigation evidenced the presence of protein Ki-67 in the nuclei of epithelial cells. In the unmodified epithelium of the oral mucosa, all epithelial cells with stained nuclei are virtually located in the basal layer. Conclusion: Against the general increase of the proliferative activity of epithelial cells with increasing SIN, a characteristic distribution of proliferating cells in the thickness of the epithelium was revealed for each studied group, as follows: in the control group and in leukoplakia without atypia, immunopositive cells are located in the basal layer, in leukoplakia (SIN1, SIN2 and SIN3 – in parabasal position while, in squamous

  2. Immunohistochemical Detection of Lipocalin 2 Specific Receptor in Psoriasis

    Directory of Open Access Journals (Sweden)

    Hamide Sayar

    2015-06-01

    Full Text Available Objective: Serum Lipocalin 2 (LCN2 levels are a general indicator for increased inflammation in the patients with psoriasis. In this study, we aimed to investigate the receptor levels of LCN2 in psoriatic lesions. Methods: Immunohistochemical expression of LCN2 was investigated in 31 skin biopsy samples of patients with psoriasis and 31 skin biopsies of control group and the results were compared between the groups. Results: Among patient group, 19 patients (61.3% had plaque psoriasis and in all of these, medium to strong staining intensity of LCN2 immunohistochemical expression was detected in the upper epidermal layer. In remaining 12 patients (38.7% with pathologically early stage psoriatic lesions and control group epidermal LCN2 immunohistochemical expression was not observed. LCN2 immunohistochemical expression was detected in the inner root sheath of the hair follicles in both groups without any statistically difference (p=1.00. Conclusion: In our study, medium to strong staining of LCN2 receptor was detected in the superficial epidermis of the psoriatic plaques. But LCN2 immunohistochemical staining was not observed in control skin biopsies. These findings may point out the significance of LCN2 in pathogenesis of psoriasis. On the other hand, absence of expression in patients with early psoriatic lesions imply that LCN2 may be active at the later stages of the pathology.

  3. Acid-fast stain

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/article/003766.htm Acid-fast stain To use the sharing features on this page, please enable JavaScript. The acid-fast stain is a laboratory test that determines ...

  4. Joint fluid Gram stain

    Science.gov (United States)

    ... called a smear. Several different colored stains are applied to the sample. The laboratory personnel will look at the stained smear under a microscope to see if bacteria are present. The color, size, and shape of ...

  5. Masson Trichrome Stain Helps Differentiate Myofibroma from Smooth Muscle Lesions in the Head and Neck Region

    Directory of Open Access Journals (Sweden)

    Julia Yu Fong Chang

    2008-10-01

    Conclusion: The Masson trichrome stain can be a useful tool to differentiate myofibromas from smooth muscle lesions, but immunohistochemical methods to rule out other spindle cell lesions are still needed.

  6. Nerve fibre studies in skin biopsies in peripheral neuropathies. I. Immunohistochemical analysis of neuropeptides in diabetes mellitus

    DEFF Research Database (Denmark)

    Lindberger, M; Schröder, H D; Schultzberg, M

    1989-01-01

    Standardised skin biopsies followed by immunohistochemical examination for the presence of terminal nerve fibres reacting for neuropeptides substance P (SP) and calcitonin gene-related peptide (CGRP) were evaluated. Healthy subjects regularly displayed free nerve endings of both fibre types...... in the dermis layers. Five type I diabetes patients without clinical or neurophysiological evidence of polyneuropathy also had reduced density of both fibre types, being significant for CGRP fibres when compared with controls. Skin biopsy with immunohistochemical staining for neuropeptides may represent...... in the papillary and reticular dermis. Both fibre types were present close to blood vessels, while CGRP immunoreactive fibres were more often encountered near sweat gland acini compared to SP fibres. Diabetes mellitus complicated by polyneuropathy was accompanied by marked reduction of SP and CGRP reactive fibres...

  7. Immunohistochemical findings after LASIK confirm in vitro LASIK model.

    Science.gov (United States)

    Priglinger, Siegfried G; May, Christian-Albrecht; Alge, Claudia S; Wolf, Armin; Neubauer, Aljoscha S; Haritoglou, Christos; Kampik, Anselm; Welge-Lussen, Ulrich

    2006-04-01

    To compare immunohistochemical findings in human donor corneas after successful laser in situ keratomileusis (LASIK) without clinical complications with a recently established human LASIK in vitro model. Donor corneas with prior LASIK treatment were investigated. Cryostat sections were stained immunohistochemically for collagen types I, III, and VI and laminin and fibronectin. With light microscopy, the interface of the LASIK flap could hardly be detected. In all samples, fibronectin was consistently detected along the entire extent of the surgical wound. In contrast, collagen type III and laminin only stained the superficial portion of the LASIK incision site. Staining for collagen types I and VI showed no changes after LASIK. Histologic findings in donor corneas with prior LASIK treatment confirm histologic observations in a recently introduced human organ culture LASIK model. This strengthens the reliability of the latter LASIK model for further studies concerning wound healing after LASIK surgery.

  8. Automatic quantification of IHC stain in breast TMA using colour analysis.

    Science.gov (United States)

    Fernández-Carrobles, M Milagro; Bueno, Gloria; García-Rojo, Marcial; González-López, Lucía; López, Carlos; Déniz, Oscar

    2017-11-01

    Immunohistochemical (IHC) biomarkers in breast tissue microarray (TMA) samples are used daily in pathology departments. In recent years, automatic methods to evaluate positive staining have been investigated since they may save time and reduce errors in the diagnosis. These errors are mostly due to subjective evaluation. The aim of this work is to develop a density tool able to automatically quantify the positive brown IHC stain in breast TMA for different biomarkers. To avoid the problem of colour variation and make a robust tool independent of the staining process, several colour standardization methods have been analysed. Four colour standardization methods have been compared against colour model segmentation. The standardization methods have been compared by means of NBS colour distance. The use of colour standardization helps to reduce noise due to stain and histological sample preparation. However, the most reliable and robust results have been obtained by combining the HSV and RGB colour models for segmentation with the HSB channels. The segmentation provides three outputs based on three saturation values for weak, medium and strong staining. Each output image can be combined according to the type of biomarker staining. The results with 12 biomarkers were evaluated and compared to the segmentation and density calculation done by expert pathologists. The Hausdorff distance, sensitivity and specificity have been used to quantitative validate the results. The tests carried out with 8000 TMA images provided an average of 95.94% accuracy applied to the total tissue cylinder area. Colour standardization was used only when the tissue core had blurring and fading stain and the expert could not evaluate them without a pre-processing. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Dramatic Stained Glass.

    Science.gov (United States)

    Prater, Michael

    2002-01-01

    Describes an art project that is appropriate for students in fifth through twelfth grade in which they create Gothic-style stained-glass windows. Discusses how college students majoring in elementary education created stained-glass windows. Addresses how to adapt this lesson for younger students. (CMK)

  10. Iron Stain on Wood

    Science.gov (United States)

    Mark Knaebe

    2013-01-01

    Iron stain, an unsightly blue–black or gray discoloration, can occur on nearly all woods. Oak, redwood, cypress, and cedar are particularly prone to iron stain because these woods contain large amounts of tannin-like extractives. The discoloration is caused by a chemical reaction between extractives in the wood and iron in steel products, such as nails, screws, and...

  11. Joint Variable Selection and Classification with Immunohistochemical Data

    Directory of Open Access Journals (Sweden)

    Debashis Ghosh

    2009-01-01

    Full Text Available To determine if candidate cancer biomarkers have utility in a clinical setting, validation using immunohistochemical methods is typically done. Most analyses of such data have not incorporated the multivariate nature of the staining profiles. In this article, we consider modelling such data using recently developed ideas from the machine learning community. In particular, we consider the joint goals of feature selection and classification. We develop estimation procedures for the analysis of immunohistochemical profiles using the least absolute selection and shrinkage operator. These lead to novel and flexible models and algorithms for the analysis of compositional data. The techniques are illustrated using data from a cancer biomarker study.

  12. Digital simulation of staining in histopathology multispectral images: enhancement and linear transformation of spectral transmittance

    Science.gov (United States)

    Bautista, Pinky A.; Yagi, Yukako

    2012-05-01

    Hematoxylin and eosin (H&E) stain is currently the most popular for routine histopathology staining. Special and/or immuno-histochemical (IHC) staining is often requested to further corroborate the initial diagnosis on H&E stained tissue sections. Digital simulation of staining (or digital staining) can be a very valuable tool to produce the desired stained images from the H&E stained tissue sections instantaneously. We present an approach to digital staining of histopathology multispectral images by combining the effects of spectral enhancement and spectral transformation. Spectral enhancement is accomplished by shifting the N-band original spectrum of the multispectral pixel with the weighted difference between the pixel's original and estimated spectrum; the spectrum is estimated using M Masson's trichrome stained equivalent show the viability of the method.

  13. Immunohistochemical biomarkers in ameloblastomas.

    Science.gov (United States)

    do Canto, Alan Motta; Rozatto, Juliana Rodrigues; Schussel, Juliana Lucena; de Freitas, Ronaldo Rodrigues; Hasséus, Bengt; Braz-Silva, Paulo Henrique

    2016-11-01

    Ameloblastoma is an aggressive odontogenic tumour, which is locally invasive and highly recurrent. Studies show that ameloblastoma is a benign odontogenic neoplasia, being relatively rare and occasionally presenting behaviour of malignant lesions. In addition to these particularities, the histological diagnosis of ameloblastoma can be challenging when the tumour shows high rates of mitosis, absence of nuclear pleomorphism, basilar hyperplasia and neural invasion. In order to help in the diagnosis, prognosis and treatment of this neoplasia, some immunohistochemical markers were shown to be associated with tumoural epithelium. The identification of these markers as well as of their association with clinical signs can be useful to elaborate more efficient treatment strategies and to control this pathology, including improvement of the quality of life of patients affected by this neoplasia. This article aims to review some markers associated with specific molecular pathways, bone remodelling, cell proliferation, apoptosis, cell signalling and tumour suppression.

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

    Science.gov (United States)

    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

  15. Stain Removal Assessment of Two Manual Toothbrushes with an Interproximal Tooth Stain Index.

    Science.gov (United States)

    Farrell, Svetlana; Grender, Julie M; Terézhalmy, Geza; Archila, Luis R

    2015-01-01

    To assess a newly developed index to measure interproximal stain and evaluate the stain removal efficacy of two commercially available manual toothbrushes. This was a randomized, examiner-blind, parallel-group, two-treatment clinical trial of two weeks' duration. Subjects qualified for the study if they had an average Modified Lobene Stain Index of ≥ 1.5 from two anterior teeth. At baseline, subjects brushed in front of a mirror for one minute under supervision. All subjects were provided with a standard 0.243% sodium fluoride dentifrice and were randomly assigned either an Oral-B Pulsar manual brush (OBP) or a Colgate Whitening manual brush (CW) to use for two weeks. Stain was reassessed after two weeks of product use. Stain measurements were conducted using the Modified Lobene Stain Index and the new Interproximal Modified Lobene Stain Index, which allows for assessment of stain in hard-to-reach areas using the same area and intensity scales as the Modified Lobene Stain Index. Use of the two manual brushes resulted in statistically significant reductions in surface stain relative to baseline after two weeks of use. Median stain reductions were 78% and 60% for the OBP and CW, respectively, as measured by the Modified Lobene Stain Index. The mean changes in the composite scores from baseline to week two were 1.85 and 1.57 for the two treatment groups, respectively. Statistically significant reductions from baseline were also found for the intensity and extent of stain measures (p brush and 83% reduction with the CW brush. For the gingival sites, the median stain removal percentages were 83% and 50%, respectively For the body region, a median stain removal of 100% was found for both treatment groups. No statistically significant differences were found between the two groups for the mean composite scores for either index. Both manual brushes showed effective stain removal, including interproximal hard-to-reach sites. The Interproximal Modified Lobene Stain Index

  16. Immunohistochemical expression of CD56 in dog (Canis familiaris) odontogenesis.

    Science.gov (United States)

    Nel, Sulette; van Heerden, Marlene; van Heerden, Willie

    2015-10-01

    To investigate the expression of CD56 in dog odontogenesis in order to elucidate the expression found in ameloblastomas. Immunohistochemical analysis of CD56 expression of developing dog teeth in the bud, cap and bell stages including the remnants of the dental lamina. Weak CD56 expression was observed in the dental epithelium during the bud stage with intense staining of certain peripheral epithelial cells. Positive staining of epithelial cells was also observed in the cap stage with intense staining of the inner enamel epithelium at this stage. During the bell stage the staining was concentrated on the cervical loop areas. The dental papilla revealed positive staining throughout the cap and bell stages while the dental follicle stained intensely positive throughout all the phases examined. The dental lamina and Serres rests also stained positive for CD56. The expression of CD56 in dog odontogenic tissue varies according to the stage of tooth development. There is a positive correlation between the positive staining observed in ameloblastomas and their odontogenic cells of origin. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. System-wide immunohistochemical analysis of protein co-localization.

    Directory of Open Access Journals (Sweden)

    Minjung Kim

    Full Text Available The analysis of co-localized protein expression in a tissue section is often conducted with immunofluorescence histochemical staining which is typically visualized in localized regions. On the other hand, chromogenic immunohistochemical staining, in general, is not suitable for the detection of protein co-localization. Here, we developed a new protocol, based on chromogenic immunohistochemical stain, for system-wide detection of protein co-localization and differential expression.In combination with a removable chromogenic stain, an efficient antibody stripping method was developed to enable sequential immunostaining with different primary antibodies regardless of antibody's host species. Sections were scanned after each staining, and the images were superimposed together for the detection of protein co-localization and differential expression. As a proof of principle, differential expression and co-localization of glutamic acid decarboxylase67 (GAD67 and parvalbumin proteins was examined in mouse cortex.All parvalbumin-containing neurons express GAD67 protein, and GAD67-positive neurons that do not express parvalbumin were readily visualized from thousands of other neurons across mouse cortex. The method provided a global view of protein co-localization as well as differential expression across an entire tissue section. Repeated use of the same section could combine assessments of co-localization and differential expression of multiple proteins.

  18. Histochemical and Immunohistochemical Study of Peripolar Cells in Sheep

    OpenAIRE

    Passos Joana; Prada Justina; Bento Lígia; Rodrigues Paula; Pires Isabel

    2013-01-01

    Peripolar cells are granulated cells located in the vascular pole of the renal corpuscle. Even though these cells have already been described, there are still many unknown histological and physiological characteristics. We carried out histochemical and immunohistochemical analyses of peripolar cells in sheep and compared their number in both normal and injured kidneys, discriminating according to the age of the animal. We tested HE, Toluidine Blue, PAS, and Masson's Trichrome stains to selec...

  19. Clinicopathologic and immunohistochemical profile of ovarian metastases from colorectal carcinoma

    OpenAIRE

    Kir, Gozde; Gurbuz, Ayse; Karateke, Ates; Kir, Mustafa

    2010-01-01

    Metastasis of colorectal adenocarcinoma of the ovary is not an uncommon occurrence and ovarian metastases from colorectal carcinoma frequently mimic endometrioid and mucinous primary ovarian carcinoma. The clinical and pathologic features of metastatic colorectal adenocarcinoma involving the ovary is reviewed with particular focus on the diagnostic challenge of distinguishing these secondary ovarian tumors from primary ovarian neoplasm. Immunohistochemical stains that may be useful in the dif...

  20. Immunohistochemical sweat gland profiles.

    Science.gov (United States)

    Noël, Fanchon; Piérard, Gérald E; Delvenne, Philippe; Quatresooz, Pascale; Humbert, Philippe; Piérard-Franchimont, Claudine

    2013-09-01

    Human sweat glands are heterogeneous in their structures and functions. Accordingly, eccrine, apocrine, and apoeccrine glands are distinguished. Some immunohistochemical markers are expected to distinguish the sweat gland types in their secretory and excretory parts. This study used two sets of antibodies. The first panel was composed of antibodies directed to well-defined sweat gland structures. The molecular targets included the low-molecular-weight cytokeratins CAM 5.2, the S100-B protein, the epithelial membrane antigen (EMA), the carcinoembryonic antigen (CEA), and the lectin Ulex europaeus agglutinin-1 (UEA-1). A second exploratory panel of antibodies targeted syndecan-1 (CD138), NKI-C3 (CD63), and CD68. They were used to disclose some undescribed antigen expressions in human sweat glands. The first set of antibodies confirmed previous findings. The immunoreactivities of the three sweat gland types were similar in the excretory ducts. By contrast, they were distinguished in the deeper coiled secretory portions of the glands. Clues supporting their distinction and probably their functional activity were obtained by immunohistochemistry using the S100-B protein, CEA and CD63 antibodies. The immunoreactivity to the S100-B protein, CEA and CD63 possibly help identifying apoeccrine sweat glands or a peculiar functional activity of eccrine sweat glands. © 2013 Wiley Periodicals, Inc.

  1. Fluorescent Europium Chelate Stain

    Science.gov (United States)

    Scaff, W. L.; Dyer, D. L.; Mori, K.

    1969-01-01

    The europium chelate of 4,4,4-trifluoro-1-(2-thienyl)-1,3-butanedione (thenoyl-trifluoroacetone; TTA) is firmly bound to microorganisms. It fluoresces brightly at 613 nm with activation at 340 nm. Cells may be stained with 10−3m chelate in 50% ethyl alcohol, followed by washing with 50% ethyl alcohol. Equal or better stains are produced with 10−3m aqueous europium salt, water wash, and 10−2m aqueous TTA. A noncomplexing buffer should be used to maintain the pH at 6.5 to 6.8. Images PMID:4181107

  2. Immunohistochemical characterization of human olfactory tissue.

    Science.gov (United States)

    Holbrook, Eric H; Wu, Enming; Curry, William T; Lin, Derrick T; Schwob, James E

    2011-08-01

    The pathophysiology underlying human olfactory disorders is poorly understood because biopsying the olfactory epithelium (OE) can be unrepresentative and extensive immunohistochemical analysis is lacking. Autopsy tissue enriches our grasp of normal and abnormal olfactory immunohistology and guides the sampling of the OE by biopsy. Furthermore, a comparison of the molecular phenotype of olfactory epithelial cells between rodents and humans will improve our ability to correlate human histopathology with olfactory dysfunction. An immunohistochemical analysis of human olfactory tissue using a comprehensive battery of proven antibodies. Human olfactory mucosa obtained from 21 autopsy specimens was analyzed with immunohistochemistry. The position and extent of olfactory mucosa was assayed by staining whole mounts (WMs) with neuronal markers. Sections of the OE were analyzed with an extensive group of antibodies directed against cytoskeletal proteins and transcription factors, as were surgical specimens from an esthesioneuroblastoma. Neuron-rich epithelium is always found inferior to the cribriform plate, even at advanced age, despite the interruptions in the neuroepithelial sheet caused by patchy respiratory metaplasia. The pattern of immunostaining with our antibody panel identifies two distinct types of basal cell progenitors in human OE similar to rodents. The panel also clarifies the complex composition of esthesioneuroblastoma. The extent of human olfactory mucosa at autopsy can easily be delineated as a function of age and neurologic disease. The similarities in human versus rodent OE will enable us to translate knowledge from experimental animals to humans and will extend our understanding of human olfactory pathophysiology. Copyright © 2011 The American Laryngological, Rhinological, and Otological Society, Inc.

  3. [Histopathological and immunohistochemical features of cardiac myxomas].

    Science.gov (United States)

    Hernández-Bringas, Omar; Ortiz-Hidalgo, Carlos

    2013-01-01

    Mixomas are the most common primary cardiac tumors with an estimate incidence of 0,5-1 per 10(6) individuals per year. These tumors have generated interest due to their unique location (left side of the atrial septum near the fossa ovalis), variable clinical presentation and undefined histogenesis. Most cardiac myxomas occur sporadically while approximately 10% of diagnosed cases develop as part of Carney complex. This neoplasm is of uncertain histogenesis, however, endothelial, neurogenic, fibroblastic, and cardiac and smooth muscle cells differentiation has been proposed, and rarely glandular differentiation has been observed. Recently, due to the expression of certain cardiomyocyte-specific factors, an origin of mesenchymal cardiomyocytes progenitor cells has been suggested. Histologically cardiac myxomas are mainly composed of stellated, fusiform and polygonal cells, immersed in an amorphous myxoid matrix. Immunohistochemically some endothelial markers, such as CD31, CD34, FVIIIAg, are present. Positive staining has also been reported for S-100 protein, calretinin, vimentin, desmin, smooth muscle myosin, CD56, α1 antitrypsin and α 1antichymotrypsin. Surgical resection is currently the only treatment of choice. We present in this article a histopathological and immunohistochemical review of cardiac myxomas. Copyright © 2012 Instituto Nacional de Cardiología Ignacio Chávez. Published by Masson Doyma México S.A. All rights reserved.

  4. Automated robust registration of grossly misregistered whole-slide images with varying stains

    Science.gov (United States)

    Litjens, G.; Safferling, K.; Grabe, N.

    2016-03-01

    Cancer diagnosis and pharmaceutical research increasingly depend on the accurate quantification of cancer biomarkers. Identification of biomarkers is usually performed through immunohistochemical staining of cancer sections on glass slides. However, combination of multiple biomarkers from a wide variety of immunohistochemically stained slides is a tedious process in traditional histopathology due to the switching of glass slides and re-identification of regions of interest by pathologists. Digital pathology now allows us to apply image registration algorithms to digitized whole-slides to align the differing immunohistochemical stains automatically. However, registration algorithms need to be robust to changes in color due to differing stains and severe changes in tissue content between slides. In this work we developed a robust registration methodology to allow for fast coarse alignment of multiple immunohistochemical stains to the base hematyoxylin and eosin stained image. We applied HSD color model conversion to obtain a less stain color dependent representation of the whole-slide images. Subsequently, optical density thresholding and connected component analysis were used to identify the relevant regions for registration. Template matching using normalized mutual information was applied to provide initial translation and rotation parameters, after which a cost function-driven affine registration was performed. The algorithm was validated using 40 slides from 10 prostate cancer patients, with landmark registration error as a metric. Median landmark registration error was around 180 microns, which indicates performance is adequate for practical application. None of the registrations failed, indicating the robustness of the algorithm.

  5. Immunohistochemical expression of Bcl-2 and Ki-67 in oral lichen planus and leukoplakia with different degrees of dysplasia.

    Science.gov (United States)

    Pigatti, Fernanda Mombrini; Taveira, Luís Antônio de Assis; Soares, Cléverson Teixeira

    2015-02-01

    Oral lichen planus (OLP) is a chronic inflammatory disease of unknown cause. Malignant transformation in OLP lesions may be favored by changes in the expression of proteins that regulate cell proliferation and apoptosis. This study aimed to investigate these issues by immunohistochemical staining for Bcl-2 and Ki-67 and by correlating histopathological findings in samples from lesions of OLP and leukoplakia with epithelial dysplasia. Data for patients with OLP or leukoplakia with moderate or severe epithelial dysplasia recorded during 2006-2011 were retrospectively reviewed. The study samples represented 37 subjects with OLP (n = 14), leukoplakia with moderate (n = 8) or severe (n = 6) epithelial dysplasia, and normal buccal mucosa (controls, n = 9). New sections were subjected to histological examination and immunohistochemistry for Bcl-2 and Ki-67 in the basal layer, suprabasal layer, and inflammatory infiltrate, respectively. All basal layer sections stained either negative or positive in leukoplakia, 85.7% of sections indicated positivity in leukoplakia (64.3%) sections stained positive for Ki-67 in >50% of cells. All suprabasal sections stained either negative or positive in leukoplakia (42.9% and 57.1%, respectively), and control (88.9% and 11.1%, respectively) samples. Suprabasal staining for Ki-67 was negative or positive in leukoplakia (7.1% and 92.9%, respectively), and controls (88.9% and 11.1%, respectively). Staining for Bcl-2 in inflammatory infiltrate in OLP was positive in 92.9% of sections. Expression of Bcl-2 may play a dual role in tumor development and progression. Increased cell proliferation in the epithelium may present a predisposition to cancer in OLP. The expression of Ki-67 can be considered as an adjunct marker for proliferative activity in lesions with malignant potential. The prognostic value of these immunomarkers in the evaluation of precancerous oral lesions requires further investigation. © 2014 The International Society

  6. Shimmering Stained Glass.

    Science.gov (United States)

    Simon, Gail Murray

    1998-01-01

    Presents an art lesson for fifth- and sixth-graders where they create a translucent design of colored cellophane on black paper inspired by the stained-glass windows of the Middle Ages and the artwork of Lewis Comfort Tiffany. Enables the students to become crafts people rather than just observers of the past. (CMK)

  7. Stained Glass and Flu

    Centers for Disease Control (CDC) Podcasts

    2017-02-01

    Dr. Robert Webster, an Emeritus member of the Department of Infectious Diseases at St. Jude Children's Research Hospital, discusses his cover art story on stained glass and influenza.  Created: 2/1/2017 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 2/1/2017.

  8. Localization of actin in normal human hepatocytes using fluorescent phallotoxins and immunohistochemical amplification.

    Science.gov (United States)

    Benkoel, L; Brisse, J; Capo, C; Benoliel, A M; Bongrand, P; Garcia, T; Chamlian, A

    1992-07-01

    Two different methods, fluorescent phallotoxins and immunohistochemical amplification systems were used to visualize actin in normal human hepatocytes. With fluorescent phallotoxins (NBD-phallacidin or rhodamine phalloidin), F-actin was distributed along the plasma membranes and at the bile canaliculi. With immunohistochemical methods (biotin-avidin, biotin-streptavidin, silver enhancement), actin was found at the same level, however a cytoplasmic staining was observed and discussed as G-actin localization.

  9. Ovarian microcystic stromal tumor: report of a new entity with immunohistochemical and ultrastructural studies.

    Science.gov (United States)

    Yang, Mary; Bhattacharjee, Meenakshi B

    2014-08-01

    Microcystic stromal tumor is a recently described rare subtype of ovarian tumor for which there has been no previously reported ultrastructural study. We report a case with the characteristic histological and immunohistochemical features and the first ultrastructural study. The immunohistochemical findings of strong and diffuse nuclear staining for beta catenin and P27 are suggestive of dysregulation of more than one genetic pathway. The ultrastructural findings are supportive of the previous postulation of an ovarian stromal origin of the neoplastic cells.

  10. Electron microscopic and immunohistochemical examination of scarred human cornea re-treated by excimer laser.

    Science.gov (United States)

    Bleckmann, Heinrich; Schnoy, Norbert; Kresse, Hans

    2002-04-01

    To elucidate differences, at the macromolecular level, in corneal tissue subjected to repeated argon fluoride excimer treatment. A light microscopic, electron microscopic, and immunohistochemical study was performed on a scarred human cornea. Keratocytes were enlarged with an expanded endoplasmic reticulum and exhibited a fibroblastic appearance. Amorphous material was observed extracellularly. Collagen fibrils exhibited a disordered arrangement while banding patterns and diameter were normal. Immunohistochemical investigation of several collagen types, of collagen-associated proteoglycans, and of basement membrane components demonstrated an enhanced immunoreactivity of all of them in the scarred area. Type V collagen was found as a normal component of the epithelial basement membrane whereas types I and III collagen were present beneath Bowman's layer. Excimer-laser-treated sections revealed considerably stronger subepithelial staining for collagen types I, III, IV, and V. Laminin-1, a typical component of basement membranes, was detectable throughout the scarred tissue. The small proteoglycans decorin and fibromodulin accumulated in a patch-like manner in the scarred tissue below the epithelium, whereas biglycan was expressed by the epithelium and throughout the stroma. Lumican was expressed most strongly by the epithelium and rather equally distributed in the excimer-laser-treated and in the normal stroma. Effects of argon laser treatment of the cornea must be regarded as a process acting over many months. Intra- and extracellular structures and components are involved and influence the unpredictable shape of the corneal architecture.

  11. Immunohistochemical localization of human papilloma virus in conjunctival neoplasias: A retrospective study

    Directory of Open Access Journals (Sweden)

    Sen Seema

    2007-01-01

    Full Text Available Background: The extent of association of human papilloma virus (HPV in human conjunctival neoplasias has been debated in studies originating from different parts of the world, but no substantial evidence has been generated on Indian subjects. This prompted us to carry out a retrospective study on conjunctival neoplasias diagnosed over the past 12 years. Materials and Methods: Histopathological and immunohistochemical analysis of 65 specimens of ocular neoplasias and 30 normal controls diagnosed between 1991 and 2002 at a tertiary eye care hospital, was undertaken. Formalin-fixed, paraffin-embedded tissues were reviewed for confirming histopathological diagnosis, presence of koilocytosis and changes related to actinic keratosis. Immunohistochemical analysis was done using HPV-specific monoclonal antibodies. Clinicopathological correlation and the association of HPV antigen with the histopathological features were performed. Results: Out of the 65 cases analyzed, 35 were papillomas and 30 were ocular surface squamous neoplasias (OSSN. The mean age was 48 years with a male preponderance. Histologically, koilocytosis was observed in 17.1% of papillomas and 36.6% of OSSN. Actinic keratosis was present in 33% of OSSN. Immunohistochemically 17.1% conjunctival papillomas stained positive for HPV antigen, all cases of OSSN were negative for HPV. There was no correlation between koilocytosis or actinic keratosis and the detection of HPV antigen. Conclusions: The association between HPV and conjunctival neoplasias is variable in different geographical areas and also depends on the methods of detection used. This study warrants the need for applying more advanced techniques at a molecular level to determine the possible etiology of HPV in conjunctival neoplasias among Asian-Indians.

  12. Marcação imunoistoquímica da expressão astrocitária de proteína glial fibrilar ácida e de vimentina no sistema nervoso central de cães com cinomose Immunohistochemical staining of the astrocytic expression of glial fibrillary acidic protein and vimentin in the central nervous system of dogs with canine distemper

    Directory of Open Access Journals (Sweden)

    Heloísa Orsini

    2007-12-01

    immunohistochemical staining of two astrocytic proteins - glial fibrillary acidic protein (GFAP and vimentin (VIM -, comparing samples of cerebellum and brainstem from eight dogs with canine distemper and from two healthy dogs, from different breeds and ages varying from 1 to 4 years old. Histological sections were submitted to the avidin-biotin-peroxidase indirect method of immmunohistochemical staining (ABC and the astrocytic reactivity, observed in light microscopy, was quantified in a computer system for image analysis. It was possible to notice, on most of the sections from sick animals, degenerative lesions that indicate demyelination. The immunostaining for GFAP and VIM was more intense on animals with canine distemper, specially around the ventricules and near degenerated sites. There was no significant difference between the immunostaining (GFAP and VIM of animals with canine distemper with and without inflammatory infiltrate of the cerebellar white matter. The increased immunoreactivity of astrocytes for GFAP and the VIM reexpression in injured areas indicate the astrocytic involvement on nervous tissue response to the demyelinating lesions induced by the canine distemper virus (CDV in the CNS.

  13. Hidradenoma Papilliferum With Oncocytic Metaplasia: A Histopathological and Immunohistochemical Study.

    Science.gov (United States)

    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.

  14. Detection of Entamoeba histolytica in experimentally induced amoebic liver abscess: comparison of three staining methods

    OpenAIRE

    Tan Zi Ning; Wong Weng Kin; Shaymoli Mustafa; Arefuddin Ahmed; Rahmah Noordin; Tan Gim Cheong; Olivos-Garcia Alfonso; Lim Boon Huat

    2012-01-01

    Objective: To compare the efficacy of three different tissue stains, namely haematoxylin and eosin (H&E), periodic-acid Schiff (PAS) and immunohistochemical (IHC) stains for detection of Entamoeba histolytica (E. histolytica) trophozoites in abscessed liver tissues of hamster. Methods: Amoebic liver abscess was experimentally induced in a hamster by injecting 1 × 106 of axenically cultured virulent E. histolytica trophozoites (HM1-IMSS strain) into the portal vein. After a week post-inocul...

  15. Masson Trichrome Stain Helps Differentiate Myofibroma from Smooth Muscle Lesions in the Head and Neck Region

    OpenAIRE

    Chang, Julia Yu Fong; Kessler, Harvey P.

    2008-01-01

    Myofibromas are well described in the head and neck region, but differentiating them from smooth muscle lesions is still difficult using smooth muscle immunohistochemical stains. This study evaluated the usefulness of the Masson trichrome stain in differentiating myofibromas from smooth muscle lesions in the head and neck region. Methods: Samples of 11 oral myofibromas, two leiomyomas, one angioleiomyoma, and one smooth muscle hamartoma were retrieved from our archives. Immunohistochemistr...

  16. Efficacy test of a toothpaste in reducing extrinsic dental stain

    Science.gov (United States)

    Agustanti, A.; Ramadhani, S. A.; Adiatman, M.; Rahardjo, A.; Callea, M.; Yavuz, I.; Maharani, D. A.

    2017-08-01

    This clinical trial compared the external dental stain reduction achieved by tested toothpaste versus placebo in adult patients. In this double-blind, parallel, randomised clinical trial, 45 female volunteers with a mean age of 20 years old were included. All study subjects front teeth were topically applicated with Silver Diamine Fluoride (SDF) to create external dental stains. Subjects were randomized into test (n=22) and control (n=23) groups. Toothpastes were used for two days to analyse the effects of removing external stains on the labial surfaces of all anterior teeth. VITA Easyshade Advance 4.0 was used to measure dental extrinsic stains changes. The analysis showed statistically significant efficacy of the tested toothpaste in reducing external dental stain caused by SDF, comparing to the placebo toothpaste, after one and two days of usage. The tested toothpaste was effective in reducing dental stain.

  17. Immunohistochemical evaluation of proliferation, apoptosis and steroidogenic enzymes in the ovary of rats with polycystic ovary

    Directory of Open Access Journals (Sweden)

    Leonardo Augusto Lombardi

    2014-07-01

    Full Text Available Objective: to evaluate the immunohistochemical expression of proliferative, apoptotic and steroidogenic enzyme markers in the ovaries of rats with polycystic ovary syndrome (PCOS. Methods: twenty rats were divided into two groups: GCtrl - estrous phase, and PCOS - with polycystic ovaries. The GCtrl animals were subjected to a lighting period from 7 am to 7 pm, while the animals with PCOS group remained with continuous lighting for 60 days. Subsequently, the animals were anesthetized, the ovaries were removed and fixed in 10% formaldehyde, prior to paraffin embedding. Sections were stained using H.E. or subjected to immunohistochemical methods for the detection of Ki-67, cleaved caspase-3, CYP11A1, CYP17A1 and CYP19A1. The results were analyzed using Student's t-test (p < 0,05. Results: morphological results showed evidence of interstitial cells originating from the inner theca cells of degenerating ovarian cysts in PCOS. Immunoexpression of Ki-67 was higher in the granulosa cells in GCtrl, and the theca interna cells in PCOS, while cleaved caspase-3 was higher in granulosa cells of ovarian cysts from PCOS and in the theca interna cells of GCtrl. Immunoreactivity of CYP11A1 in the theca interna, granulosa and interstitial cells was similar between the two groups, while CYP17A1 and CYP19A1 were higher in the granulosa and interstitial cells in the PCOS group. Conclusion: the results indicate that the interstitial cells are derived from the theca interna and that enzymatic changes occur in the theca interna and interstitial cells in ovaries of rats with PCOS, responsible for the high levels of androgens and estradiol.

  18. Digital simulation of staining in histopathology multispectral images: enhancement and linear transformation of spectral transmittance.

    Science.gov (United States)

    Bautista, Pinky A; Yagi, Yukako

    2012-05-01

    Hematoxylin and eosin (H&E) stain is currently the most popular for routine histopathology staining. Special and/or immuno-histochemical (IHC) staining is often requested to further corroborate the initial diagnosis on H&E stained tissue sections. Digital simulation of staining (or digital staining) can be a very valuable tool to produce the desired stained images from the H&E stained tissue sections instantaneously. We present an approach to digital staining of histopathology multispectral images by combining the effects of spectral enhancement and spectral transformation. Spectral enhancement is accomplished by shifting the N-band original spectrum of the multispectral pixel with the weighted difference between the pixel's original and estimated spectrum; the spectrum is estimated using M transformed to the spectral configuration associated to its reaction to a specific stain by utilizing an N × N transformation matrix, which is derived through application of least mean squares method to the enhanced and target spectral transmittance samples of the different tissue components found in the image. Results of our experiments on the digital conversion of an H&E stained multispectral image to its Masson's trichrome stained equivalent show the viability of the method.

  19. Masson trichrome stain helps differentiate myofibroma from smooth muscle lesions in the head and neck region.

    Science.gov (United States)

    Chang, Julia Yu Fong; Kessler, Harvey P

    2008-10-01

    Myofibromas are well described in the head and neck region, but differentiating them from smooth muscle lesions is still difficult using smooth muscle immunohistochemical stains. This study evaluated the usefulness of the Masson trichrome stain in differentiating myofibromas from smooth muscle lesions in the head and neck region. Samples of 11 oral myofibromas, two leiomyomas, one angioleiomyoma, and one smooth muscle hamartoma were retrieved from our archives. Immunohistochemistry and Masson trichrome stains were performed on tissue sections of these lesions. All 11 oral myofibromas, seven from male patients and four from female patients, were solitary myofibromas. The patients' mean age at diagnosis was 32.8 years. Oral myofibromas occurred most commonly on the gingiva (four cases) and in the mandible (three cases). With the Masson trichrome stain, the smooth muscle cell cytoplasm was stained red, while the collagenous fibrous tissue was stained blue. Myofibromas and smooth muscle lesions demonstrated different characteristic patterns with the Masson trichrome stain. Myofibromas were composed of a much more collagenous stroma intermixed with the spindle cells. Thick fibrous bundles with random, irregularly intersecting angles were prominent in myofibromas. Smooth muscle lesions showed only minimal delicate fibrous tissue surrounding the smooth muscle cells and in the septa between the smooth muscle masses. On low-power view, red masses of smooth muscle tumor surrounded by blue fibrous tissue were observed. The Masson trichrome stain can be a useful tool to differentiate myofibromas from smooth muscle lesions, but immunohistochemical methods to rule out other spindle cell lesions are still needed.

  20. Christendom's Narratives and the Stained Glass Designs of Yusuf ...

    African Journals Online (AJOL)

    Stained glass paintings, the most distinctive accessory in the interior repertoire of Christian ecclesiastic spaces, is the subject matter of this study. This paper attempts a recast of Christendom's narratives in the stained glass designs of Yusuf Cameron Adebayo Grillo as the distinctive overarching mechanism of the ...

  1. Immunohistochemical localization of androgen receptor in rat caput epididymis during postnatal development

    Directory of Open Access Journals (Sweden)

    Sema Timurkaan

    2011-09-01

    Full Text Available Objectives: The aim of this study was to investigate the developmental pattern of androgen receptor (AR in caput epididymis.Materials and methods: In this study three randomly selected rats were sacrificed at ages 21, 56, 90 and 120 days old. All male rats were anesthetized with ethyl ether before killing. Then, the caput epididymides were removed and fixed in Bouin’s fixative at +4°C for 36 hour. Afterwards the tissue samples were embedded in paraffin for routine histological methods. Later the tissues were sectioned at 5μm and mounted on poly-L-lysin-coated slides. To solve the antigen masking problem, we performed microwave stimulated antigen retrieval technique before the immunohistochemical staining. Avidin-Biotin-Peroxidase Complex (ABC method was applied for immunohistochemical staining.Results: In all age groups of rats studied, positive immunohistochemical staining for the AR appeared in nuclei of epididymal cells. The staining intensity of AR positive cells did not change depending on age. In caput epididymis, immunostainable AR was found in tubular epithelial cells (principal cells, basal cells and apical cells and peritubular smooth muscle cells. The AR staining in the epithelial cells appeared to be stronger than in the peritubular smooth muscle cells. In the epithelial cells; staining intensity was stronger in principal cells than in basal cells and apical cells.Conclusion: Staining intensity of AR positive epididymal cells irrespective of age indicated the necessity of androgens for postnatal differentiation and maintaining the structure of the epididymis. Stronger staining intensity in principal cells suggested that principal cells are more sensitive to androgen stimulation. J Clin Exp Invest 2011; 2 (3: 260-266.

  2. Histological and Immunohistochemical Revision of Hepatocellular Adenomas: A Learning Experience

    Directory of Open Access Journals (Sweden)

    S. Fonseca

    2013-01-01

    Full Text Available Light has been shed on the genotype/phenotype correlation in hepatocellular adenoma (HCA recognizing HNF1α-inactivated HCA (H-HCA, inflammatory HCA (IHCA, and β-catenin-activated HCA (b-HCA. We reviewed retrospectively our surgical HCA series to learn how to recognize the different subtypes histopathologically and how to interpret adequately their immunohistochemical staining. From January 1992 to January 2012, 37 patients underwent surgical resection for HCA in our institution. Nine had H-HCA (25% characterized by steatosis and loss of L-FABP expression; 20 had IHCA (55.5% showing CRP and/or SAA expression, sinusoidal dilatation, and variable inflammation; and 1 patient had both H-HCA and IHCA. In 5 patients (14%, b-HCA with GS and β-catenin nuclear positivity was diagnosed, two already with hepatocellular carcinoma. Two cases (5.5% remained unclassified. One of the b-HCA showed also the H-HCA histological and immunohistochemical characteristics suggesting a subgroup of β-catenin-activated/HNF1α-inactivated HCA, another b-HCA exhibited the IHCA histological and immunohistochemical characteristics suggesting a subgroup of β-catenin-activated/inflammatory HCA. Interestingly, three patients had underlying vascular abnormalities. Using the recently published criteria enabled us to classify histopathologically our retrospective HCA surgical series with accurate recognition of b-HCA for which we confirm the higher risk of malignant transformation. We also underlined the association between HCA and vascular abnormalities.

  3. Reliability of a rapid hematology stain for sputum cytology

    OpenAIRE

    Gonçalves, Jéssica; Pizzichini, Emilio; Pizzichini, Marcia Margaret Menezes; Steidle, Leila John Marques; Rocha, Cristiane Cinara; Ferreira, Samira Cardoso; Zimmermann, Célia Tânia

    2014-01-01

    Objective: To determine the reliability of a rapid hematology stain for the cytological analysis of induced sputum samples. Methods: This was a cross-sectional study comparing the standard technique (May-Grünwald-Giemsa stain) with a rapid hematology stain (Diff-Quik). Of the 50 subjects included in the study, 21 had asthma, 19 had COPD, and 10 were healthy (controls). From the induced sputum samples collected, we prepared four slides: two were stained with May-Grünwald-Giemsa, and two w...

  4. Histochemical and Immunohistochemical Study of Peripolar Cells in Sheep

    Directory of Open Access Journals (Sweden)

    Passos Joana

    2013-01-01

    Full Text Available Peripolar cells are granulated cells located in the vascular pole of the renal corpuscle. Even though these cells have already been described, there are still many unknown histological and physiological characteristics. We carried out histochemical and immunohistochemical analyses of peripolar cells in sheep and compared their number in both normal and injured kidneys, discriminating according to the age of the animal. We tested HE, Toluidine Blue, PAS, and Masson's Trichrome stains to select the best stain for identification and quantification. Masson Trichrome yielded the best results and was selected for this purpose. We identified the cells by the presence of cytoplasmatic granules and by their position in the vascular pole. We found no statistically significant association between the number of peripolar cells and the age of the animal or the occurrence of lesions. In the immunohistochemical analysis, we found that the cells were positive to α-smooth muscle actin and less consistently positive to NSE and S100 protein. Chromogranin A, cyclooxygenase-2, AE1/AE3, and Wide Spectrum Cytokeratin and desmin yielded negative results. We conclude that although there was evidence of a contractile function, there was no evidence to support that peripolar cells have either a neuroendocrine or an epithelial nature.

  5. Immunohistochemical and molecular characterization of the human periosteum.

    Science.gov (United States)

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

    2013-01-01

    The aim of the present study was to characterize the cell of the human periosteum using immunohistological and molecular 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. 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. 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.

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

    Science.gov (United States)

    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. Copyright © 2014 Elsevier Ltd. All rights reserved.

  7. Successful treatment of granuloma faciale with topical tacrolimus: a case report and immunohistochemical study.

    Science.gov (United States)

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

    2012-05-01

    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 Foxp3(high+) 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.

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

    Directory of Open Access Journals (Sweden)

    Gen-ichi Tojo

    2012-07-01

    Full Text Available 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.

  9. Microwave oven-based technique for immunofluorescent staining of paraffin-embedded tissues.

    Science.gov (United States)

    Long, Delwin J; Buggs, Colleen

    2008-02-01

    Immunohistochemical analysis of formalin-fixed paraffin-embedded tissues can be challenging due to potential modifications of protein structure by exposure to formalin. Heat-induced antigen retrieval techniques can reverse reactions between formalin and proteins that block antibody recognition. Interactions between antibodies and antigens are further enhanced by microwave irradiation, which has simplified immunohistochemical staining protocols. In this report, we modify a technique for antigen retrieval and immunofluorescent staining of formalin-fixed paraffin-embedded tissues by showing that it works well with several antibodies and buffers. This microwave-assisted method for antigen retrieval and immunofluorescent staining eliminates the need for blocking reagents and extended washes, which greatly simplifies the protocol allowing one to complete the analysis in less than 3 h.

  10. Role of bcl-2 oncoprotein in oral potentially malignant disorders and squamous cell carcinoma: An immunohistochemical study

    Directory of Open Access Journals (Sweden)

    V M Sudha

    2011-01-01

    Full Text Available Background and Objectives: The product of bcl-2 gene, bcl-2 protein, an anti-apoptotic protein, is known to be over-expressed in potentially malignant disorders and squamous cell carcinoma (SCC of the oral cavity. The aim of this study is to compare the topographical aspect and degree of bcl-2 over-expression in potentially malignant disorders including leukoplakia, oral submucous fibrosis (OSMF, and oral lichen planus (OLP, with that of the oral squamous cell carcinoma (OSCC, and to determine whether bcl-2 protein can be considered as a tumor marker. Materials and Methods : A group of 60 histo-pathologically diagnosed, formalin-fixed, paraffin embedded tissue samples was included in the study. The study group was further subdivided into four groups: Group I, consisting of oral leukoplakia; Group II, OSMF; Group III, OLP and Group IV, OSCC. These samples were collected from Government Dental College, Bangalore, and then subjected to immunohistochemical (IHC staining using indirect immunoenzyme labeled streptavidin biotin (LSAB method. Results : Out of 30 cases of OSCC: 11 (36.7% cases showed greater supra-basal keratinocyte staining; 15 (50% cases showed greater number of positive cells in the basal cell layer, with relatively less number of supra-basal cells showing positive staining; and, rest of the 4 (13.3% cases did not show convincing staining. Among the total 30 cases of potentially malignant disorders: 10 each of leukoplakia, OSMF and OLP, 2 (20%, 2 (20%, 4 (40% of the cases showed greater supra-basal cell layer positive staining and 8 (80%, 6 (60%, 6 (60% of them showed greater basal cell staining, respectively. Two cases of OSMF did not show convincing staining. In the cases that were bcl-2 positive: 2 (6.67% of the OSCC, 3 (30% of leukoplakia, 2 (20% of OSMF and 1 (10% of OLP, showed more than 50% of the cells positive. 25-50% cells were positive in 21 (70% of OSCC, 6 (60% of leukoplakia, 4 (40% of OSMF and 6 (60% of OLP cases. 10

  11. The Social Function of Staining

    OpenAIRE

    GÜNDÜZ, Alev

    2016-01-01

    The place of staining which is established in today’s cosmetic perception as a creative way forbeauty, charm and attraction, had been split the path of the fed formatting of the social necessity ofthe past. The adventure of staining person who constantly reshapes the area based for needing, initiallypointed to similar meanings in the mind and body of every individual in the society. A personwho finds self-expression using non-verbal superior language by staining; has created a new sourceof id...

  12. Histopathological, immunohistochemical and special stain unraveling the enigmatic carcinosarcoma - A case report

    Directory of Open Access Journals (Sweden)

    Moidin Shakil

    2015-01-01

    Full Text Available Carcinosarcoma is an extremely rare, highly malignant tumor having a characteristic of both epithelial and mesenchymal components; the epithelial component exhibiting focal squamous cell carcinoma and mesenchymal component having sarcomatoid stroma. The occurrence of carcinosarcoma in the oral cavity is very rare and only few cases have been reported in the literature, although its occurrence in other parts of the body has been widely reported.

  13. Immunohistochemical staining for the differentiation of subungual keratoacanthoma from subungual squamous cell carcinoma.

    LENUS (Irish Health Repository)

    Connolly, M

    2008-08-01

    Subungual keratotic tumours are rare. The clinical and histological distinctions between subungual keratoacanthomas (SUKAs) and subungual squamous cell carcinomas (SCCs) are important, but often difficult. Adequate methods of differentiation between the two are required, both for the purpose of management and for assessment of prognosis.

  14. Immunohistochemical staining of human sperm cells in smears from sexual assault cases

    DEFF Research Database (Denmark)

    Christoffersen, S.

    2011-01-01

    In the routine clinical examination of sexual assault victims, apart from documenting physical evidence of abuse, securing evidence, typically DNA from blood, semen, or saliva, is an important part of the process. Often the presence of semen is considered a most interesting piece of evidence....... Not only does it often contain enough DNA for DNA profiling, but it also strongly indicates that an actual sexual act has taken place. The examination of smear slides obtained in sexual assault cases is a time-consuming task especially for the less trained and in cases where the smear only contains few...

  15. Immunohistochemical and immunocytochemical findings associated with Marek's disease virus in naturally infected laying hens.

    Science.gov (United States)

    Yavuz, O; Erer, H

    2017-09-14

    We compared immunohistochemical (IHC) staining of tissue sections of liver, kidney, spleen, lung, proventriculus, sciatic nerve, bursa of Fabricius, brain, heart, intestine and skin; immunocytochemical (ICC) staining of peripheral blood samples and touch preparations of liver, spleen and kidney of laying hens naturally infected with Marek's disease (MD) virus. We used one hundred and fifty 5-17-week-old commercial hens. IHC and ICC staining were performed using polymer-based techniques. IHC staining exhibited mostly free immunopositive reactions in tumor cells and in the cytoplasm of the parenchymal cells of liver, kidney, spleen and bursa of Fabricius. In the sciatic nerve, severe reactions were observed in the cytoplasm of plasma and MD cells in the lymphoproliferative areas. Pronounced staining was found in the lymphoid cells in the medulla of intrafollicular regions in the bursa of Fabricius. Although immunostaining was observed in the liver and spleen touch preparations, there was no staining in the kidneys and peripheral blood cell samples. The presence of virus in the tissue and peripheral blood samples and in touch preparations was compared immunohistochemically and immunocytochemically. IHC and ICC techniques were helpful for diagnosis of MD. Peripheral blood samples are inappropriate for field conditions and natural infections.

  16. Out damn spot!--a study of the removal of dental material stains from cloth.

    Science.gov (United States)

    Cunningham, J L; Bolas, A

    1995-01-01

    Stains to white cotton caused by clinical dental materials were subjected to washing and dry cleaning processes. Most stains were successfully removed with the exception of a siloxane impression polymer.

  17. Morphological and immunohistochemical characterisation of seminomas in Norwegian dogs

    Directory of Open Access Journals (Sweden)

    Thorvaldsen Tor

    2012-09-01

    Full Text Available Abstract Background Seminomas in the dog have traditionally been assumed to resemble human spermatocytic seminomas, based on their low malignancy and high occurrence in old individuals. However, recently published studies indicate that canine seminomas can be classified as classical and spermatocytic seminomas in a similar way as in man, and that classical seminomas comprise a substantial proportion of seminomas in the dog. These two factors both contribute to increasing the potential of canine seminoma as a relevant model for human testicular cancer. The aim of the present study was to characterise seminoma in Norwegian dogs using morphology and immunohistochemistry, and determine whether these tumours are comparable with human classical seminoma. Methods By applying diagnostic criteria from human pathology, 45 seminomas from the Norwegian Canine Cancer Register were examined histologically with hematoxylin and eosin (HE and periodic acid-Schiff (PAS stains. All sections were stained immunohistochemically with antibodies against human placental alkaline phosphatase (PLAP and the transmembrane receptor c-KIT. Results Although two of the seminomas showed immunohistochemical staining characteristics indicative of classical seminoma (PLAP+/c-KIT+, all 45 examined seminomas were morphologically consistent with spermatocytic seminoma. Conclusions The value of canine seminoma as a model for SE in man remains unclear. Among the 45 investigated tumours from Norwegian dogs, none were classified as classical seminoma based on morphological criteria consistent with human seminomas. Regional or breed differences in the occurrence of classical seminoma in the dog, as well as the lack of uniform diagnostic criteria, might explain the discrepancy between the findings in the current study and the results presented by other authors.

  18. Gram stain of skin lesion

    Science.gov (United States)

    Skin lesion gram stain ... skin sore. This procedure is called a skin lesion biopsy . Before the biopsy, your provider will numb ... means bacteria have been found in the skin lesion. Further tests are needed to confirm the results. ...

  19. Clinical studies to determine the effectiveness of a whitening toothpaste at reducing stain (using a forced stain model).

    Science.gov (United States)

    Moran, J; Claydon, N C A; Addy, M; Newcombe, R

    2005-02-01

    Two single centre, randomized single-blind, crossover studies were performed, to compare the effect of a test toothpaste with a conventional fluoride paste in the inhibition and removal of extrinsic dental stain promoted by repeated chlorhexidine/tea rinses. These studies used 24 subjects in each of two separate clinical trials. On the Friday before each trial period, the subjects received a prophylaxis to remove all staining, plaque and calculus deposits. On the following Monday, subjects were checked whether they were stain free and then under direct supervision they rinsed with a 0.2% chlorhexidine mouthrinse, immediately followed by a rinse with a warm black tea solution. This cycle was repeated hourly eight times throughout the day and on the following days until the Friday. In addition subjects also received daily a single toothpaste slurry rinse or control water rinse in the morning and lunchtime. No other form of oral hygiene was permitted during this period. On the Friday, both stain area and intensity was assessed using the Lobene Stain Index. For the stain removal study, stain was promoted again using chlorhexidine and tea rinses. After 4 days, stain was measured both prior to and immediately after brushing with the allocated toothpaste for 2 min. Subjects were then instructed to use the toothbrush at home according to their normal oral hygiene practices. On the following Wednesday, the amount of stain present was re-assessed. Each subject subsequently received a thorough prophylaxis to remove all plaque calculus and staining before starting the following periods of the study. The study showed no difference in the ability of the test whitening toothpaste, control toothpaste and water control at inhibiting stain. There was also only a small difference (3.5% for product of area and intensity) between the ability of the two toothpastes to help remove stain after a single brushing. The difference was however in favour of the test product which approached a

  20. Sebaceous carcinoma: an immunohistochemical reappraisal.

    Science.gov (United States)

    Ansai, Shin-ichi; Takeichi, Hiromi; Arase, Seiji; Kawana, Seiji; Kimura, Tetsunori

    2011-08-01

    The rates of distant metastases and tumor death in sebaceous carcinoma (SC) have been reported to be higher than those of other cutaneous carcinomas, such as squamous cell carcinoma (SCC) and basal cell carcinoma (BCC), regardless of whether they occur in ocular or extraocular regions. Therefore, strict differentiation of SC from SCC and BCC is required. In this article, we report immunohistochemical findings of SC and compare these data to those of SCC, BCC, and sebaceoma. An immunohistochemical study was performed using 7 antibodies [anti-carcinoembryonic antigen (CEA), anti-epithelial membrane antigen (EMA), anti-CA15-3, anti-CA19-9, anti-androgen receptor (AR), anti-epithelial antigen (Ber-EP4), and anti-adipophilin (ADP)] on 35 cases of SC (16 cases in ocular and 19 cases in extraocular regions) and 10 cases of each SCC (5 cases in ocular and 5 cases in extraocular regions), BCC (5 cases in ocular and 5 cases in extraocular regions), and sebaceoma (no cases arose on the eyelids). In summary, the typical immunophenotypes of SC were EMA+, CA15-3+, AR+, Ber-EP4-, and ADP+; those of sebaceoma were CEA-, EMA+, Ber-EP4-, and ADP+; those of SCC were CEA-, EMA+, CA19-9-, AR-, Ber-EP4-, and ADP-; and those of BCC were CEA-, EMA-, CA15-3-, Ber-EP4+, and ADP-. Other antibody tests for each neoplasm were positive in about half of the cases. The detection of AR and ADP was useful for differentiating SC from SCC, whereas the determination of EMA, CA15-3, Ber-EP4, and ADP was valuable in differentiating SC from BCC.

  1. Ductal adenocarcinoma of the prostate: immunohistochemical findings and clinical significance

    Directory of Open Access Journals (Sweden)

    Sha JJ

    2013-10-01

    Full Text Available Jianjun Sha,1,2 Juanjie Bo,1 Jiahua Pan,1 Lianhua Zhang,1 Hanqing Xuan,1 Wei Chen,1 Dong Li,1 Zhaoliang Wang,1 Dongming Liu,1 Yiran Huang1,2 1Department of Urology, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, 2School of Biomedical Engineering, Shanghai Jiaotong University, Shanghai, People's Republic of China Introduction: To investigate the clinical features, diagnosis, treatment, and prognosis of ductal adenocarcinoma of the prostate. Methods: The clinicopathological and immunohistochemical data of seven patients with ductal adenocarcinoma of the prostate were retrospectively analyzed. All patients underwent physical examination, magnetic resonance imaging (MRI, bone scan, cystoscopy, and computed tomography (CT scan. The level of prostate-specific antigen (PSA before and after surgery was assessed. Different prostate cancer markers were used for immunohistochemical staining. Results: The mean age of the seven patients diagnosed with prostatic ductal adenocarcinoma in this study was 76.2 years (range 57–88. Five patients presented with intermittent and painless gross hematuria, one patient with progressive dysuria, and one patient with elevated serum PSA on routine health examination. The level of PSA before surgery ranged from 1.3 to 45.0 ng/mL. Immunohistochemical staining results of the prostatic ductal adenocarcinoma confirmed positivity for PSA, prostatic acid phosphatase, androgen receptor, and alpha-methyacyl co-enzyme A (CoA-reductase markers. Two of the patients underwent bilateral orchiectomy combined with anti-androgen therapy, three underwent transurethral resection of prostate, one received radical prostatectomy, and one received medical castration therapy. The clinical outcomes of all patients were satisfactory, based on follow-up data. The symptoms of hematuria and dysuria were ameliorated well, and the postoperative PSA level decreased below 4.0 ng/mL. Recurrence or metastasis of disease was

  2. Meningeal Hemangiopericytomas and Meningomas: a Comparative Immunohistochemical and Genetic Study.

    Science.gov (United States)

    Trabelsi, Saoussen; Mama, Nadia; Chourabi, Maroua; Mastouri, Maroua Haddaji; Ladib, Mohamed; Popov, Sergey; Burford, Anna; Mokni, Moncef; Tlili, Kalthoum; Krifa, Hedi; Jones, Chris; Yacoubi, Mohamed Tahar; Saad, Ali; Brahim, Dorra H'mida-Ben

    2015-01-01

    The meningeal hemangiopericytoma (MHPC) is a vascular tumor arising from pericytes. Most intracranial MHPCs resemble meningiomas (MNGs) in their clinical presentation and histological features and may therefore be misdiagnosed, despite important differences in prognosis. We report 8 cases of MHPC and 5 cases of MNG collected from 2007 to 2011 from the Neuro-Surgery and Histopathology departments. All 13 samples were re reviewed by two independent pathologists and investigated by immunohistochemistry (IHC) using mesenchymal, epithelial and neuro-glial markers. Additionally, we screened all tumors for a large panel of chromosomal alterations using multiplex ligation probe amplification (MLPA). Presence of the NAB2-STAT6 fusion gene was inferred by immunohistochemical staining for STAT6. Compared with MNG, MHPCs showed strong VIM (100% of cases), CD99 (62%), bcl-2 (87%), and p16 (75%) staining but only focal positivity with EMA (33%) and NSE (37%). The p21 antibody was positive in 62% of MHPC and less than 1% in all MNGs. MLPA data did not distinguish HPC from MNG, with PTEN loss and ERBB2 gain found in both. By contrast, STAT6 nuclear staining was observed in 3 MHPC cases and was absent from MNG. MNG and MHPC comprise a spectrum of tumors that cannot be easily differentiated based on histopathology. The presence of STAT6 nuclear positivity may however be a useful diagnostic marker.

  3. Expression of type I and type III collagens in oral submucous fibrosis: An immunohistochemical study

    Directory of Open Access Journals (Sweden)

    Venkatesh V Kamath

    2015-01-01

    Full Text Available Background: Oral submucous fibrosis (OSMF is a potentially malignant collagen - metabolic disorder linked to consumption of betel quid and areca nut. The deposition of collagen and its major subtypes have been the subject of intense scrutiny in the etiopathogenesis of the disorder. Aims and Objectives: The present study was planned to immunohistochemically identify the expression of collagens I and III (COL I and III in different grades of OSF and compare it with normal oral mucosa and scar tissue. Materials and Methods: Archival paraffin sections of 72 cases of various grades of OSMF, ten cases of normal mucosa as controls and four cases of scar tissue were stained with antibodies to COL I and III (BioGenex Laboratories, CA, USA to evaluate the collagen subtypes on paraffin sections. The expression was quantified by image analysis software (Jenoptik Optical System, ProgRes ® Capture Pro, version 2.8.8 and statistically analyzed. Results: COL I and III stained all the tissues ubiquitously. COL I was more in ratio and quantity in all the grades of OSMF, normal mucosa, and scar tissue. The proportion of COL I to COL III seemed to increase with progressive grades of OSF. Interestingly, during the process of fibrosis COL III seems to be deposited earlier and gradually replaced by COL I resulting in a skewed ratio vis a vis normal oral mucosa and scar tissue. Conclusions: COL I expression was found to be proportionate with advancing grades of OSF while COL III expression increased in Grade I but subsequently decreased as severity of OSF increased. The increase in COL I at the expense of COL III showed a similar pattern in the submucosa while in the deeper muscle only Grade III cases reflected the trend. While all cases of OSF revealed excessive expression in comparison with normal oral mucosa, the comparison with scar tissue was variable.

  4. Histopathological and immunohistochemical aspects of American cutaneous leishmaniasis before and after different treatments.

    Science.gov (United States)

    Viana, Agostinho Gonçalves; Mayrink, Wilson; Fraga, Carlos Alberto de Carvalho; Silva, Luciana Maria; Domingos, Patrícia Luciana Batista; Bonan, Paulo Rogério Ferreti; Paula, Alfredo Maurício Batista de; Botelho, Ana Cristina de Carvalho

    2013-01-01

    The histopathology and immune responses of the healing process of leishmaniasis are still poorly studied. This study aimed to examine the histopathological and immunological aspects of lesions of patients with cutaneous leishmaniasis before and after different therapeutic methods. We studied 23 individuals grouped according to the treatments: Glucantime, Glucantime + Leishvacin and Glucantime + Leishvacin associated with Bacillus Calmette-Guerin. For analysis of the histopathological changes present in the dermis and epidermis, histological sections were stained with hematoxylin and eosin. The samples were immunostained before and after treatment to analyze the expression of interferon (IFN)-γ, interleukin (IL) 12, IL-10 and IL-4. Before treatment the presence of intense infiltrates of mononuclear cells was noticed and after treatment, even with a diagnosis of clinical cure, the subjects still showed a moderate inflammatory process. In the immunohistochemical analyses, we noticed a difference between the cytokines, with increased expression of cytokines IFN-γ and IL-12 compared to IL 10 and IL-4, both before and after treatment and, comparatively, the difference in this expression was more intense before treatment. However, the cytokine expression analyzed by treatment group showed no statistically significant difference. We conclude that a clinical cure does not always coincide with the histopathological one, and that before treatment there is a predominance of Th1 cytokines. In terms of treatment type, there was no difference in the progression of healing for all the three types of treatment, indicating their clinical equivalence.

  5. Histopathological and immunohistochemical aspects of American cutaneous leishmaniasis before and after different treatments*

    Science.gov (United States)

    Viana, Agostinho Gonçalves; Mayrink, Wilson; Fraga, Carlos Alberto de Carvalho; Silva, Luciana Maria; Domingos, Patrícia Luciana Batista; Bonan, Paulo Rogério Ferreti; de Paula, Alfredo Maurício Batista; Botelho, Ana Cristina de Carvalho

    2013-01-01

    BACKGROUND The histopathology and immune responses of the healing process of leishmaniasis are still poorly studied. OBJECTIVES This study aimed to examine the histopathological and immunological aspects of lesions of patients with cutaneous leishmaniasis before and after different therapeutic methods. METHODS We studied 23 individuals grouped according to the treatments: Glucantime, Glucantime + Leishvacin and Glucantime + Leishvacin associated with Bacillus Calmette-Guerin. For analysis of the histopathological changes present in the dermis and epidermis, histological sections were stained with hematoxylin and eosin. The samples were immunostained before and after treatment to analyze the expression of interferon (IFN)-γ, interleukin (IL) 12, IL-10 and IL-4. RESULTS Before treatment the presence of intense infiltrates of mononuclear cells was noticed and after treatment, even with a diagnosis of clinical cure, the subjects still showed a moderate inflammatory process. In the immunohistochemical analyses, we noticed a difference between the cytokines, with increased expression of cytokines IFN-γ and IL-12 compared to IL 10 and IL-4, both before and after treatment and, comparatively, the difference in this expression was more intense before treatment. However, the cytokine expression analyzed by treatment group showed no statistically significant difference. CONCLUSION We conclude that a clinical cure does not always coincide with the histopathological one, and that before treatment there is a predominance of Th1 cytokines. In terms of treatment type, there was no difference in the progression of healing for all the three types of treatment, indicating their clinical equivalence. PMID:23539001

  6. Occurrence and significance of Mallory bodies in morbidly obese patients. An immunohistochemical study

    DEFF Research Database (Denmark)

    Gluud, C; Christoffersen, Pernille Yde; Andersen, T

    1984-01-01

    Liver biopsies from 61 consecutive patients with morbid obesity (less than 60% overweight) and from 48 patients with alcoholic liver disease were examined for the presence of Mallory bodies. For the detection both routine haematoxylin and eosin stained sections and sections exposed...... liver disease. Using the immunohistochemical technique, Mallory bodies were found in the liver of 2 obese patients (3%) and in 36 patients with alcoholic liver disease (75%). None of the Mallory body positive obese patients showed signs of diabetes mellitus, cholestasis or hypocholesterolemia, but both...... patients admitted previous excessive alcohol consumption. It is concluded that the immunohistochemical detection of Mallory bodies is more sensitive than routine staining. Further, Mallory bodies are rare findings in livers of obese patients and may be related to excessive alcohol consumption....

  7. Absence of lymphatic vessels in the dog dental pulp: an immunohistochemical study.

    Science.gov (United States)

    Martin, Anna; Gasse, Hagen; Staszyk, Carsten

    2010-11-01

    In spite of numerous investigations it has not been precisely determined whether lymphatic vessels are present in the dental pulp of dogs. Therefore, this study attempted a specific immunohistochemical detection of lymphatic endothelium. The canine teeth of 19 healthy beagle dogs were dissected into three segments (apical, intermediate and occlusal). After decalcification, specimens were embedded in paraffin wax and histologic cross-sections were stained immunohistochemically using a reliable antibody (anti-Prox-1) against the homeobox transcription factor Prox-1, which is located within the nucleus of lymphatic endothelium. Anti-Prox-1 reacted positively with canine control tissues (lymph nodes, gingiva, nasal mucosa), but showed no staining in tissue sections of the dental pulp. The dog dental pulp contained no vascular structures lined with lymphatic endothelium. This suggests that drainage of interstitial fluid makes use of other routes, i.e. extravascular pathways. © 2010 The Authors. Journal of Anatomy © 2010 Anatomical Society of Great Britain and Ireland.

  8. Immunohistochemical study of the expression of cell cycle regulating proteins at different stages of bladder cancer

    DEFF Research Database (Denmark)

    Primdahl, Hanne; von der Maase, Hans; Sørensen, Flemming Brandt

    2002-01-01

    ; group 1, 23 patients with recurrent stage Ta (non-invasive) tumors; group 2, 22 patients presenting at their first admission with T2-4 (muscle invasive) tumors; group 3, 24 patients who experienced disease progression from Ta or T1 (invasive in connective tissue) to a higher stage......PURPOSE: The cell cycle is known to be deregulated in cancer. We therefore analyzed the expression of the cell cycle related proteins p21, p27, p16, Rb, and L-myc by immunohistochemical staining of bladder tumors.METHODS: The tissue material consisted of bladder tumors from three groups of patients....... By immunohistochemical staining the protein expression was compared to allelic deletions of the corresponding genes. The allelic deletions were detected by PCR-based microsatellite analyses.RESULTS: We detected a significant reduction in the expression levels of the cell cycle related proteins p21(waf1) ( P=0.002), p27...

  9. Rare Giant Granular Cell Ameloblastoma: A Case Report and an Immunohistochemical Study

    Directory of Open Access Journals (Sweden)

    Santosh Hunasgi

    2013-01-01

    Full Text Available Aims. The aim is to present a case of rare giant granular cell ameloblastoma and to review the pertinent literature highlighting the molecular aspects of its pathogenesis by analyzing the expression of CD-68, Bcl-2, and β-catenin. Methods. H and E stained sections showed large odontogenic islands showing peripheral ameloblast-like cells and central stellate reticulum-like cells with extensive granular cell transformation surrounded by fibrous stroma. Polyclonal rabbit anti-CD 68, anti-Bcl2, and anti-β-catenin were stained immunohistochemically. Results. CD-68 showed a moderate to strong staining intensity in granular cells. Moderate staining of Bcl-2 was expressed by the peripheral columnar cells of tumor islands and negative in the granular cells. Expression of β-catenin was generally weak, except for only the focal areas that showed a moderate staining intensity and weak in peripheral cells. Conclusion. The present case of giant granular cell ameloblastoma is a rare entity. Development of monstrous size is indicative of ameloblastomas persistent growth. Granular cell transformation in ameloblastomas probably occurs as a consequence of extensive molecular changes. Immunohistochemical studies help us to know the pathogenesis of this granular cell ameloblastoma. Therefore, an effort has been made here to study the expression of Bcl-2, CD-68, and β-catenin.

  10. Detection of Entamoeba histolytica in experimentally induced amoebic liver abscess: comparison of three staining methods.

    Science.gov (United States)

    Ning, Tan Zi; Kin, Wong Weng; Mustafa, Shaymoli; Ahmed, Arefuddin; Noordin, Rahmah; Cheong, Tan Gim; Alfonso, Olivos-Garcia; Huat, Lim Boon

    2012-01-01

    To compare the efficacy of three different tissue stains, namely haematoxylin and eosin (H&E), periodic-acid Schiff (PAS) and immunohistochemical (IHC) stains for detection of Entamoeba histolytica (E. histolytica) trophozoites in abscessed liver tissues of hamster. Amoebic liver abscess was experimentally induced in a hamster by injecting 1 × 10(6) of axenically cultured virulent E. histolytica trophozoites (HM1-IMSS strain) into the portal vein. After a week post-inoculation, the hamster was sacrificed and the liver tissue sections were stained with H&E, PAS and IHC stains to detect the amoebic trophozoite. The three stains revealed tissue necrosis and amoebic trophozoites, but with varying clarity. H&E and PAS stained the trophozoites pink and magenta, respectively, however it was difficult to differentiate the stained trophozoites from the macrophages because of their similarity in size and morphology. On the other hand, IHC stain revealed distinct brown appearance of the trophozoites in the infected liver tissues. It can be concluded that out of the three stains, IHC is the best for identification of E. histolytica trophozoites in tissue sections.

  11. Special stains in Mohs surgery.

    Science.gov (United States)

    Miller, Christopher J; Sobanko, Joseph F; Zhu, Xiaodong; Nunnciato, Terri; Urban, Christopher R

    2011-04-01

    The excellent cure rates associated with Mohs micrographic surgery depend on accurate interpretation of complete and high-quality microscopic frozen sections. Reliable interpretation of microscopic slides is only possible if the surgeon can distinguish tumor cells from surrounding normal tissue. By highlighting tumor cells with a chromogen that is visible on light microscopy, immunostaining allows the Mohs surgeon to distinguish tumor from normal cells in these challenging scenarios. This article focuses on practical aspects involving the most commonly used immunostains in dermatologic surgery, including MART-1 for melanocytic neoplasms, cytokeratin stains for keratinocytic neoplasms, and CD34 stains for dermatofibrosarcoma protuberans. Copyright © 2011 Elsevier Inc. All rights reserved.

  12. [Lugol staining for esophageal carcinoma and influence of radiotherapy].

    Science.gov (United States)

    Miyamoto, H; Adachi, W; Koike, S; Koide, N; Iida, F

    1993-01-01

    For evaluating the endoscopic staining of the esophageal carcinoma with lugol solution, 50 patients who underwent esophagectomy for carcinoma were subjected to this study. Among the 50 patients, 21 were radiated before surgery and 29 were not radiated. The findings of the lugol staining were compared between endoscopic staining and staining on removed specimens. Non-staining area demonstrated by endoscopic procedure almost agreed with that by the procedure on removed specimen in non-radiation group, but both areas of 28.6% cases disagreed in radiation group. On the second step, the extent of non-staining area demonstrated by the procedure of removed specimen was compared with histological extent of carcinoma. The non-staining area on the removed specimen was more extended than histological extent of carcinoma; 10.3% in the non-radiation group and 71.4% in the radiation group. As one of the causes of the large non-corresponding rate in the radiation group, radiation esophagitis was demonstrated. It can be finally concluded that the reliability of endoscopic lugol staining is reduced by preoperative irradiation.

  13. Modified elastic tissue-Masson trichrome stain.

    Science.gov (United States)

    Garvey, W

    1984-07-01

    A combined elastic tissue-Masson technique is presented which stains elastic fibers of all sizes, nuclei and connective tissue. The modified elastic tissue stain consists of hematoxylin, ferric chloride and Verhoeff's iodine; nuclei and elastic fibers are stained blue-black in six minutes without differentiation. By contrast, cytoplasmic elements are stained red, (Biebrich scarlet-acid fuchsin) and collagen is stained green (light green) or blue (aniline blue). The entire staining procedure takes approximately one hour.

  14. Immunohistochemical expression of phosphatase and tensin homolog in histologic gradings of oral squamous cell carcinoma

    Directory of Open Access Journals (Sweden)

    Shiny S.R. Jasphin

    2016-01-01

    Full Text Available Context: Phosphatase and tensin homolog (PTEN is a tumor suppressor gene located on chromosome 10q23. PTEN has its major function in the regulation of cell adhesion, cell cycle arrest, migration, apoptosis programming, and differentiation. This genomic region suffers loss of heterozygosity in many human cancers. Aims: The aim of this study was to compare the immunohistochemical expression of PTEN in normal oral mucosa and oral squamous cell carcinoma (OSCC and to correlate the PTEN expression in gradings of OSCC. Materials and Methods: Thirty cases of paraffin tissue sections of previously diagnosed OSCC were taken. Of thirty cases, ten were well differentiated, ten were moderately differentiated, and ten were poorly differentiated. As a control, ten paraffin sections of oral normal mucosa tissue specimens were taken from patients undergoing extractions. The sections were stained for immunohistochemical expression of PTEN. The cells stained by PTEN antibody were counted, and an immunohistochemical score was obtained. Statistical Analysis Used: Statistical analysis was done using Mann–Whitney's test and Kruskal–Wallis test. Results: Statistical analysis revealed that there was a significant difference between normal mucosa and OSCC in immunohistochemistry staining. However, there was no significant difference in PTEN expression among gradings of OSCC. Conclusions: The study concluded that there was a decrease in PTEN expression in OSCC than normal mucosa. It also concluded that PTEN is a tumor suppressor gene which has a wide role in oral carcinogenesis.

  15. Accelerated staining technique using kitchen microwave oven.

    Science.gov (United States)

    Mukunda, Archana; Narayan, T V; Shreedhar, Balasundhari; Shashidhara, R; Mohanty, Leeky; Shenoy, Sadhana

    2015-01-01

    Histopathological diagnosis of specimens is greatly dependent on good sample preparation and staining. Both of these processes is governed by diffusion of fluids and dyes in and out of the tissue, which is the key to staining. Diffusion of fluids can be accelerated by the application of heat that reduces the time of staining from hours to the minute. We modified an inexpensive model of kitchen microwave oven for staining. This study is an attempt to compare the reliability of this modified technique against the tested technique of routine staining so as to establish the kitchen microwave oven as a valuable diagnostic tool. Sixty different tissue blocks were used to prepare 20 pairs of slides for 4 different stains namely hematoxylin and eosin, Van Gieson's, 0.1% toluidine blue and periodic acid-Schiff. From each tissue block, two bits of tissues were mounted on two different slides. One slide was stained routinely, and the other stained inside a microwave. A pathologist evaluated the stained slides and the results so obtained were analyzed statistically. Microwave staining considerably cut down the staining time from hours to seconds. Microwave staining showed no loss of cellular and nuclear details, uniform-staining characteristics and was of excellent quality. The cellular details, nuclear details and staining characteristics of microwave stained tissues were better than or equal to the routine stained tissue. The overall quality of microwave-stained sections was found to be better than the routine stained tissue in majority of cases.

  16. Salt stains from evaporating droplets

    NARCIS (Netherlands)

    Shahidzadeh, N.; Schut, M.F.L.; Desarnaud, J.; Prat, M.; Bonn, D.

    2015-01-01

    The study of the behavior of sessile droplets on solid substrates is not only associated with common everyday phenomena, such as the coffee stain effect, limescale deposits on our bathroom walls, but also very important in many applications such as purification of pharmaceuticals, deicing of

  17. Amelogenin in odontogenic cysts and tumors: An immunohistochemical study

    Science.gov (United States)

    Anigol, Praveen; Kamath, Venkatesh V.; Satelur, Krishnanand; Anand, Nagaraja; Yerlagudda, Komali

    2014-01-01

    Background: Amelogenins are the major enamel proteins that play a major role in the biomineralization and structural organization of enamel. Aberrations of enamel-related proteins are thought to be involved in oncogenesis of odontogenic epithelium. The expression of amelogenin is possibly an indicator of differentiation of epithelial cells in the odontogenic lesions. Aims and Objectives: The present study aimed to observe the expression of amelogenin immunohistochemically in various odontogenic lesions. Materials and Methods: Paraffin sections of 40 odontogenic lesions were stained immunohistochemically with amelogenin antibodies. The positivity, pattern and intensity of expression of the amelogenin antibody were assessed, graded and statistically compared between groups of odontogenic cysts and tumors. Results: Almost all the odontogenic lesions expressed amelogenin in the epithelial component with the exception of an ameloblastic carcinoma. Differing grades of intensity and pattern were seen between the cysts and tumors. Intensity of expression was uniformly prominent in all odontogenic lesions with hard tissue formation. Statistical analysis however did not indicate significant differences between the two groups. Conclusion: The expression of amelogenin antibody is ubiquitous in odontogenic tissues and can be used as a definitive marker for identification of odontogenic epithelium. PMID:25937729

  18. Etika Berbusana Mahasiswa Stain Samarinda

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    Ida Suryani Wijaya

    2012-06-01

    Full Text Available Ethics is about behavior of human being, such as which one is right or wrong. The ethics is always affecting the human life. The ethics gives people orientation how he/she do manything every time every day. Islamic ethics consists of the way how someone interact each other; how someone should do or not to do, how to sit, how to walk, how to eat or drink, how to sleep, or how to get dressed. Al-Qur’an uses three terms to define about dressing, they are: libas, tsiyah, and sarahi. Dressing has a function as covering the body, as assessoris, as the way to do Islamic taqwa, and as an identiy. Dressing ethics of the female students of STAIN Samarinda has been regulated by the rector regulation No 19 of the year 2002 about relation and dressing ethics for the students of STAIN Samarinda.

  19. A retrospective study of the clinical, histological, and immunohistochemical manifestations of 5 dogs originally diagnosed histologically as necrotizing scleritis.

    Science.gov (United States)

    Denk, Nora; Sandmeyer, Lynne S; Lim, Christine C; Bauer, Bianca S; Grahn, Bruce H

    2012-03-01

    To describe the clinical, histological, and immunohistochemical manifestations of canine necrotizing scleritis. A retrospective examination of the clinical records and samples of ocular tissues from five dogs with a histological diagnosis 'necrotizing scleritis' was completed. Archived, formalin-fixed, paraffin-embedded samples and two control globes were stained with hematoxylin and eosin, Gram, periodic acid-Schiff (PAS) and Masson trichrome stains, and they were immunohistochemically labeled for CD3, CD18, and CD20. Of the five cases reviewed, only two could be confirmed as idiopathic necrotizing scleritis. The other three cases were retrospectively diagnosed as unilateral focal, non-necrotizing scleritis, one as episcleritis and the third was scleritis secondary to a proptosed globe based on our retrospective clinical, histological, and immunohistochemical evaluations. In these two cases, idiopathic necrotizing scleritis manifested as a bilateral, progressive, inflammatory disease of the sclera and cornea that induces significant uveitis. Light microscopic examination confirmed collagen degeneration and granulomatous inflammation. There was no evidence for an infectious etiology based on Gram's and PAS stainings. Immunohistochemical labeling revealed a predominance of B cells in idiopathic, bilateral necrotizing scleritis. Tinctorial staining abnormalities with Masson's trichrome stain were present in scleral collagen of the two cases with idiopathic necrotizing scleritis as well as a case of secondary traumatic scleritis. Based on a limited number of cases, idiopathic canine necrotizing scleritis shares similar histopathological features with non-necrotizing scleritis and episcleritis; however, necrotizing scleritis is B-cell-dominated and bilateral, and significant collagen alterations manifest with Masson's trichrome stain. © 2011 American College of Veterinary Ophthalmologists.

  20. Immunohistochemical profiles of claudin-3 in primary and metastatic prostatic adenocarcinoma

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    Becich Michael J

    2011-01-01

    Full Text Available Abstract Background Claudins are integral membrane proteins that are involved in forming cellular tight junctions. One member of the claudin family, claudin-3, has been shown to be overexpressed in breast, ovarian, and pancreatic cancer. Here we use immunohistochemistry to evaluate its expression in benign prostatic hyperplasia (BPH, prostatic intraepithelial neoplasia (PIN, normal tissue adjacent to prostatic adenocarcinoma (NAC, primary prostatic adenocarcinoma (PCa, and metastatic prostatic adenocarcinoma (Mets. Methods Tissue microarrays were immunohistochemically stained for claudin-3, with the staining intensities subsequently quantified and statistically analyzed using a one-way ANOVA with subsequent Tukey tests for multiple comparisons or a nonparametric equivalent. Fifty-three cases of NAC, 17 cases of BPH, 35 cases of PIN, 107 cases of PCa, and 55 cases of Mets were analyzed in the microarrays. Results PCa and Mets had the highest absolute staining for claudin-3. Both had significantly higher staining than BPH (p Conclusions To our knowledge, this represents one of the first studies comparing the immunohistochemical profiles of claudin-3 in PCa and NAC to specimens of PIN, BPH, and Mets. These findings provide further evidence that claudin-3 may serve as an important biomarker for prostate cancer, both primary and metastatic, but does not provide evidence that claudin-3 can be used to predict risk of metastasis.

  1. Pathological and immunohistochemical study of lethal primary brain stem injuries

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    Rongchao Sun

    2012-05-01

    Full Text Available Abstract Background Many of the deaths that occur shortly after injury or in hospitals are caused by mild trauma. Slight morphological changes are often found in the brain stems of these patients during autopsy. The purpose of this study is to investigate the histopathological changes involved in primary brain stem injuries (PBSI and their diagnostic significance. Methods A total of 65 patients who had died of PBSI and other conditions were randomly selected. They were divided into 2 groups, an injury group (25 cases and a control group (20 cases. Slides of each patient’s midbrain, pons, and medulla oblongata were prepared and stained with HE, argentaffin, and immunohistochemical agents (GFAP, NF, amyloid-ß, MBP. Under low power (×100 and NF staining, the diameter of the thickest longitudinal axon was measured at its widest point. Ten such diameters were collected for each part of the brain (midbrain, pons, and medulla oblongata. Data were recorded and analyzed statistically. Results Brain stem contusions, astrocyte activity, edema, and pathological changes in the neurons were visibly different in the injury and control groups (P P  Conclusions These histopathological changes may prove beneficial to the pathological diagnosis of PBSI during autopsy. The measurement of axon diameters provides a referent quantitative index for the diagnosis of the specific causes of death involved in PBSI. Virtual Slides The virtual slide(s for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1345298818712204

  2. Immunohistochemical Analysis of Vimentin in Oral Submucous Fibrosis

    Science.gov (United States)

    Nayak, Meghanand T.; Singh, Anjali; Desai, Rajiv S.; Vanaki, S. S.

    2013-01-01

    Background. Oral submucous fibrosis (OSF), a precancerous condition, is characterized by abnormal accumulation of collagen fibers in oral submucosa. Vimentin is a Class 2 intermediate filament (IF) and primarily expressed in cells of mesenchymal origin. Vimentin is also found to be involved in cell growth, cell cycling, and tumour differentiation. Objective. The purpose of the study was to compare the expression of vimentin in various histological grades of OSF. Materials and Methods. To assess the immunohistochemical expression of vimentin in 20 mild cases of OSF, 20 severe cases of OSF, and ten cases of normal oral buccal mucosa. Results. The overall staining intensity of vimentin significantly increased statistically (P < 0.01) in OSF cases over normal control. A significant increase in the staining intensity of vimentin was also noted in the fibroblasts of severe cases of OSF (P = 0.03). Conclusion. Considering the marked vimentin expression in the present study, future studies should include cytoskeleton IF and other filaments in the fibroblasts of OSF. PMID:23840210

  3. Immunohistochemical Analysis of Vimentin in Oral Submucous Fibrosis

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    Meghanand T. Nayak

    2013-01-01

    Full Text Available Background. Oral submucous fibrosis (OSF, a precancerous condition, is characterized by abnormal accumulation of collagen fibers in oral submucosa. Vimentin is a Class 2 intermediate filament (IF and primarily expressed in cells of mesenchymal origin. Vimentin is also found to be involved in cell growth, cell cycling, and tumour differentiation. Objective. The purpose of the study was to compare the expression of vimentin in various histological grades of OSF. Materials and Methods. To assess the immunohistochemical expression of vimentin in 20 mild cases of OSF, 20 severe cases of OSF, and ten cases of normal oral buccal mucosa. Results. The overall staining intensity of vimentin significantly increased statistically (P<0.01 in OSF cases over normal control. A significant increase in the staining intensity of vimentin was also noted in the fibroblasts of severe cases of OSF (P=0.03. Conclusion. Considering the marked vimentin expression in the present study, future studies should include cytoskeleton IF and other filaments in the fibroblasts of OSF.

  4. Expression of human telomerase reverse transcriptase protein in oral epithelial dysplasia and oral squamous cell carcinoma: An immunohistochemical study

    Science.gov (United States)

    Raghunandan, Bangalore Nagarajachar; Sanjai, Karpagaselvi; Kumaraswamy, Jayalakshmi; Papaiah, Lokesh; Pandey, Bhavna; Jyothi, Bellur MadhavaRao

    2016-01-01

    Background: Telomerase is an RNA-dependent DNA polymerase that synthesizes TTAGGG telomeric DNA sequences and almost universally provides the molecular basis for unlimited proliferative potential. The telomeres become shorter with each cycle of replication and reach a critical limit; most cells die or enter stage of replicative senescence. Telomere length maintenance by telomerase is required for all the cells that exhibit limitless replicative potential. It has been postulated that reactivation of telomerase expression is necessary for the continuous proliferation of neoplastic cells to attain immortality. Use of immunohistochemistry (IHC) is a useful, reliable method of localizing the human telomerase reverse transcriptase (hTERT) protein in tissue sections which permits cellular localization. Although there exists a lot of information on telomerase in oral cancer, little is known about their expression in oral epithelial dysplasia and their progression to oral squamous cell carcinoma (OSCC) compared to normal oral mucosa. This study addresses this lacuna. Aims: To compare the expression of hTERT protein in oral epithelial dysplasia and OSCC with normal oral mucosa by Immunohistochemical method. Subjects and Methods: In this preliminary study, IHC was used to detect the expression of hTERT protein in OSCC (n = 20), oral epithelial dysplasia (n = 21) and normal oral mucosa (n = 10). The tissue localization of immunostain, cellular localization of immunostain, nature of stain, intensity of stain, percentage of cells stained with hTERT protein were studied. A total number of 100 cells were counted in each slide. Statistical Analysis: All the data were analyzed using SPSS software version 16.0. The tissue localization, cellular localization of cytoplasmic/nuclear/both of hTERT stain, staining intensity was compared across the groups using Pearson's Chi-square test. The mean percentage of cells stained for oral epithelial dysplasia, OSCC and normal oral mucosa were

  5. Immunohistochemical distinction of ocular sebaceous carcinoma from basal cell and squamous cell carcinoma.

    Science.gov (United States)

    Sinard, J H

    1999-06-01

    Diagnosis of sebaceous carcinoma of the periorbital region is often delayed. Clinically, this lesion can mimic several inflammatory disorders. Histopathologically, it can mimic either squamous cell or basal cell carcinoma. To identify an immunohistochemical approach to assist in the diagnosis of periorbital sebaceous carcinoma. The immunohistochemical profiles of several cases of periorbital sebaceous, basal cell, and squamous cell carcinoma were examined. Although at least focal epithelial membrane antigen (EMA) staining can effectively distinguish sebaceous carcinoma (10 of 11 were positive) from basal cell carcinoma (1 of 16 were positive), most squamous cell carcinomas examined were also focally EMA positive (11 of 14). However, Cam 5.2 reactivity was seen in most sebaceous carcinomas (8 of 11) but no squamous cell carcinomas (0 of 14). In addition, at least focal BRST-1 reactivity was also seen in most sebaceous carcinomas (7 of 11) but no basal cell carcinomas (0 of 16). Periorbital sebaceous, basal cell, and squamous cell carcinomas have different immunohistochemical staining profiles; a panel of commonly available antibodies, including anti-EMA, BRST-1, and Cam 5.2, may help distinguish these diseases from each other when that distinction cannot be clearly made by light microscopy alone.

  6. Immunohistochemical expression of growth factors in the follicular wall of normal and cystic ovaries of sows.

    Science.gov (United States)

    Sant'Ana, F J F; Reis Junior, J L; Blume, G R; Gimeno, E J; Rey, F; Ortega, H H

    2015-04-01

    The expression of growth factors was evaluated immunohistochemically in normal and cystic ovaries of sows. The immunohistochemically stained area (IHCSA) was quantified by image analysis to analyse the expression of these proteins in the follicular wall of secondary, tertiary and cystic follicles. IGF-I immunoreactivity was strong in the granulosa cell layer (GC), moderate in the theca interna (TI) and mild in the theca externa (TE) of the normal follicles. There was severe reduction of the labelling to IGF-I in the GC of the follicular and luteinized cysts. In the normal follicles, the reactivity for IGF-II was very similar to pattern noted in IGF-I. There was reduction of the IHCSAs in the GC of the follicular and luteinized cysts, but the decrease was not significant. The staining of the IGF-II in the TI and TE of the cysts was increased, in comparison with normal follicles. The IHCSAs for VEGF were higher in the GC and TE of the normal follicles in contrast to TI, but this difference was noted only in the tertiary follicle. The VEGF reactivity increased in the GC of the cysts, in relation to normal follicles. The results of the current study show that the formation of ovarian cysts in sows is associated with alterations in the immunohistochemical expression of some growth factors. © 2015 Blackwell Verlag GmbH.

  7. Immunohistochemical analysis of cytochrome C oxidase facilitates differentiation between oncocytoma and chromophobe renal cell carcinoma.

    Science.gov (United States)

    Adam, Alexander C; Scriba, Alexander; Ortmann, Monika; Huss, Sebastian; Kahl, Philip; Steiner, Susanne; Störkel, Stephan; Büttner, Reinhard

    2015-01-01

    In this study, immunohistochemical staining pattern of cytochrome c oxidase subunit 1 (CCO1) was investigated in the differentiation of renal oncocytoma (RO) from eosinophilic (EoC) and classic chromophobe renal cell carcinoma (ChRCC). A feature found in ChRCC/EoC but not in RO is the predominance of a perinuclear halo when stained for CCO1. In a cohort of 103 mixed cases including 44 RO, 37 classic ChRCC and 22 EoC, the diagnosis based on this immunohistochemical feature alone was consistent with the previous routine diagnosis in 95.7%. We reached 100% specificity and 81.4% sensitivity of this pattern in ChRCC. Specificity for RO was 93.2% and sensitivity correspondingly 95.5%. We propose a novel and easily interpretable immunohistochemical method for the discrimination of benign RO from certain subtypes of malignant ChRCC. Because of strong similarity in morphology of the 2 entities the diagnosis often cannot be made based on standard histopathology alone. The study describes for the first time the formation of a perinuclear halo in CCO1 immunohistochemistry as a highly specific marker for the diagnosis of ChRCC. We think this method can be a strong amendment for routine diagnostics in renal cell carcinoma.

  8. Basal Cell Carcinoma With Matrical Differentiation: Clinicopathologic, Immunohistochemical, and Molecular Biological Study of 22 Cases.

    Science.gov (United States)

    Kyrpychova, Liubov; Carr, Richard A; Martinek, Petr; Vanecek, Tomas; Perret, Raul; Chottová-Dvořáková, Magdalena; Zamecnik, Michal; Hadravsky, Ladislav; Michal, Michal; Kazakov, Dmitry V

    2017-06-01

    Basal cell carcinoma (BCC) with matrical differentiation is a fairly rare neoplasm, with about 30 cases documented mainly as isolated case reports. We studied a series of this neoplasm, including cases with an atypical matrical component, a hitherto unreported feature. Lesions coded as BCC with matrical differentiation were reviewed; 22 cases were included. Immunohistochemical studies were performed using antibodies against BerEp4, β-catenin, and epithelial membrane antigen (EMA). Molecular genetic studies using Ion AmpliSeq Cancer Hotspot Panel v2 by massively parallel sequencing on Ion Torrent PGM were performed in 2 cases with an atypical matrical component (1 was previously subjected to microdissection to sample the matrical and BCC areas separately). There were 13 male and 9 female patients, ranging in age from 41 to 89 years. Microscopically, all lesions manifested at least 2 components, a BCC area (follicular germinative differentiation) and areas with matrical differentiation. A BCC component dominated in 14 cases, whereas a matrical component dominated in 4 cases. Matrical differentiation was recognized as matrical/supramatrical cells (n=21), shadow cells (n=21), bright red trichohyaline granules (n=18), and blue-gray corneocytes (n=18). In 2 cases, matrical areas manifested cytologic atypia, and a third case exhibited an infiltrative growth pattern, with the tumor metastasizing to a lymph node. BerEP4 labeled the follicular germinative cells, whereas it was markedly reduced or negative in matrical areas. The reverse pattern was seen with β-catenin. EMA was negative in BCC areas but stained a proportion of matrical/supramatrical cells. Genetic studies revealed mutations of the following genes: CTNNB1, KIT, CDKN2A, TP53, SMAD4, ERBB4, and PTCH1, with some differences between the matrical and BCC components. It is concluded that matrical differentiation in BCC in most cases occurs as multiple foci. Rare neoplasms manifest atypia in the matrical areas

  9. Porcine intestinal mast cells. Evaluation of different fixatives for histochemical staining techniques considering tissue shrinkage

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    J. Rieger

    2013-07-01

    Full Text Available Staining of mast cells (MCs, including porcine ones, is critically dependent upon the fixation and staining technique. In the pig, mucosal and submucosal MCs do not stain or stain only faintly after formalin fixation. Some fixation methods are particularly recommended for MC staining, for example the fixation with Carnoy or lead salts. Zinc salt fixation (ZSF has been reported to work excellently for the preservation of fixation-sensitive antigens. The aim of this study was to establish a reliable histological method for counting of MCs in the porcine intestinum. For this purpose, different tissue fixation and staining methods that also allow potential subsequent immunohistochemical investigations were evaluated in the porcine mucosa, as well as submucosa of small and large intestine. Tissues were fixed in Carnoy, lead acetate, lead nitrate, Zamboni and ZSF and stained subsequently with either polychromatic methylene blue, alcian blue or toluidine blue. For the first time our study reveals that ZSF, a heavy metal fixative, preserves metachromatic staining of porcine MCs. Zamboni fixation was not suitable for histochemical visualization of MCs in the pig intestine. All other tested fixatives were suitable. Alcian blue and toluidine blue co-stained intestinal goblet cells which made a prima facie identification of MCs difficult. The polychromatic methylene blue proved to be the optimal staining. In order to compare MC counting results of the different fixation methods, tissue shrinkage was taken into account. As even the same fixation caused shrinkage-differences between tissue from small and large intestine, different factors for each single fixation and intestinal localization had to be calculated. Tissue shrinkage varied between 19% and 57%, the highest tissue shrinkage was found after fixation with ZSF in the large intestine, the lowest one in the small intestine after lead acetate fixation. Our study emphasizes that MC counting results from

  10. Microdissection of stained archival tissue.

    Science.gov (United States)

    Gupta, S K; Douglas-Jones, A G; Morgan, J M

    1997-08-01

    In many tissues the preinvasive stage of neoplastic progression can be identified histologically as dysplasia or in situ disease. There is much interest in defining the molecular events associated with the early stages of neoplasia. Retrieval of histologically recognisable preinvasive neoplastic tissue uncontaminated by inflammatory or stromal cells is important for genetic studies using polymerase chain reaction (PCR) assay. A novel method for microdissection is described in which 10 microns sections are dewaxed, stained with haematoxylin and eosin, dried, covered with Sellotape, and the tissue cut out using a scalpel blade under direct visual control. The method is quick, eliminates problems of operator tremor, preserves the architecture of the micro-dissected tissue (for photographic documentation) and requires no special equipment. The presence of Sellotape and adhesive in the reaction mixture has no detrimental effect on the ability to extract DNA or to perform PCR.

  11. Immunohistochemical characterization of neoplastic cells of breast origin

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    Noriega MariadelasMercedes

    2012-06-01

    Full Text Available Abstract Background After skin cancer, breast cancer is the most common malignancy in women. Tumors of unknown origin account for 5-15% of malignant neoplasms, with 1.5% being breast cancer. An immunohistochemical panel with conventional and newer markers, such as mammaglobin, was selected for the detection of neoplastic cells of breast origin. The specific objectives are: 1 to determine the sensitivity and specificity of the panel, with a special emphasis on the inclusion of the mammaglobin marker, and 2 to compare immunohistochemistry performed on whole tissue sections and on Tissue Micro-Array. Methods Twenty-nine metastatic breast tumors were included and assumed as tumors of unknown origin. Other 48 biopsies of diverse tissues were selected and assumed as negative controls. Tissue Micro-Array was performed. Immunohistochemistry for mammaglobin, gross cystic disease fluid protein-15, estrogen receptor, progesterone receptor and cytokeratin 7 was done. Results Mammaglobin positive staining was observed in 10/29 cases, in 13/29 cases for gross cystic disease fluid protein-15, in 20/29 cases for estrogen receptor, in 9/29 cases for progesterone receptor, and in 25/29 cases for cytokeratin 7. Among the negative controls, mammaglobin was positive in 2/48, and gross cystic disease fluid protein-15 in 4/48. Conclusions The inclusion of MAG antibody in the immunohistochemical panel for the detection of tumors of unknown origin contributed to the detection of metastasis of breast cancer. The diagnostic strategy with the highest positive predictive value (88% included hormone receptors and mammaglobin in serial manner. Virtual slides The virtual slide(s for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1366310812718988

  12. [Diagnostic stain of helminth eggs (author's transl)].

    Science.gov (United States)

    Cerva, L

    1976-12-01

    A description is given of a diagnostic method for the staining of eggs and larvae of intestinal helminth in smears of both fresh and fixed stool samples. The contents of the eggs and larvae stain red, the background various shades of blue. The most contrasting staining was obtained with thin-walled eggs.

  13. Granuloma faciale: clinical, morphological and immunohistochemical aspects in a series of 10 patients*

    Science.gov (United States)

    Oliveira, Cristiano Claudino; Ianhez, Pedro Eugênio de Carvalho; Marques, Silvio Alencar; Marques, Mariângela Esther Alencar

    2016-01-01

    Granuloma faciale is a chronic, benign, cutaneous vasculitis with well-established clinical and morphological patterns, but with an unknown etiology. This study describes clinical and pathologic aspects of patients diagnosed with granuloma faciale. The authors analyzed demographic, clinical, morphological and immunohistochemical data from patients with a final diagnosis of granuloma faciale, confirmed between 1998 and 2012. There was a proportional and mixed inflammatory infiltrate, Grenz zones were present in almost all the samples. Immunophenotyping confirmed a higher intensity of T lymphocytes than B lymphocytes in thirteen samples, with a predominance of T CD8 lymphocytes in 64% of cases, in contrast to the literature, which indicates that the major component is T CD4 lymphocytes. All cases were positive for IgG4 but the majority (12/14) had less than 25% of stained cells. The pathogenesis of granuloma faciale remains poorly understood, making studies of morphological and immunohistochemical characterization important to better understand it. PMID:28099604

  14. Granuloma faciale: clinical, morphological and immunohistochemical aspects in a series of 10 patients.

    Science.gov (United States)

    Oliveira, Cristiano Claudino; Ianhez, Pedro Eugênio de Carvalho; Marques, Silvio Alencar; Marques, Mariângela Esther Alencar

    2016-01-01

    Granuloma faciale is a chronic, benign, cutaneous vasculitis with well-established clinical and morphological patterns, but with an unknown etiology. This study describes clinical and pathologic aspects of patients diagnosed with granuloma faciale. The authors analyzed demographic, clinical, morphological and immunohistochemical data from patients with a final diagnosis of granuloma faciale, confirmed between 1998 and 2012. There was a proportional and mixed inflammatory infiltrate, Grenz zones were present in almost all the samples. Immunophenotyping confirmed a higher intensity of T lymphocytes than B lymphocytes in thirteen samples, with a predominance of T CD8 lymphocytes in 64% of cases, in contrast to the literature, which indicates that the major component is T CD4 lymphocytes. All cases were positive for IgG4 but the majority (12/14) had less than 25% of stained cells. The pathogenesis of granuloma faciale remains poorly understood, making studies of morphological and immunohistochemical characterization important to better understand it.

  15. Immunohistochemical testing for Helicobacter Pylori existence in neoplasms of the colon

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    Yasar Nurgul

    2008-08-01

    Full Text Available Abstract Background Helicobacter pylori is a common pathogen, and its prevalence varies with socioeconomic conditions (10–80%. It has recently been recognized as a class I carcinogen in relation to gastric cancer. The aim of this study was to investigate the presence of Helicobacter pylori in neoplasms of the colon by immunohistochemical methods. Methods The polypectomy materials of 51 patients (19 male and 32 female who had undergone colonoscopic polypectomy were retrieved for retrospective examination. The endoscopic size and colonic localization of the polyps were recorded. Hematoxylin and eosin stains were evaluated according to histological type and grade of dysplasia. Biopsy stains were immunohistochemically treated with Helicobacter pylori antibodies by the streptavidine-biotin immunoperoxidase technique. Helicobacter pylori staining in the gastric mucosa was used as the control for the immunohistochemical method. Specimens were classified according to the presence of Helicobacter pylori under an optical microscope, and Helicobacter pylori positive specimens were stratified according to the respective staining pattern. Results Mean age was 61.88 ± 10.62 (40–82 years. Polyp sizes were 1.45 ± 0.92 (1–4 cm; and 25.5% of polyps were localized in the right colon, 68.6% in the left colon and 5.9% in the transverse colon. Presence of Helicobacter pylori was not correlated with localization (p > 0.05 or size of the polyps (p > 0.05. Eleven (21.6% of all specimens included in the study were Helicobacter pylori positive by immunohistochemical methods. Of the Helicobacter pylori positive specimens, the staining pattern was diffuse: Equivocal in 90.9%, nonspecific with a finely granular type concentrated on the luminal surface in 90.9%, dot-like granular in 54.5%, and spiral in 9.1%. Of the tubular polyps, 17.9% were H. pylori positive, and the staining pattern was equivocal in 100%, luminal in 85.7%, and dot-like granular in 57.1%. Of the

  16. Extrinsic Stain Removal Effectiveness of a New Whitening Dentifrice.

    Science.gov (United States)

    Ghassemi, A; Vorwerk, L; Hooper, W; Cirigliano, A; DeSciscio, P; Nathoo, S

    2015-01-01

    This study was conducted to evaluate the effectiveness of Arm & Hammer (A&H) Truly Radiant Rejuvenating toothpaste in removing extrinsic tooth stain compared to that of a conventional fluoride/silica-containing dentifrice. This was a randomized, examiner-blind, parallel-design study with two groups of subjects who brushed unsupervised with their assigned dentifrice for two minutes, twice daily, for five days. Extrinsic stain was measured on the labial surfaces of the eight incisor teeth by the Modified Lobene Stain Index (MLSI) at baseline and following five days of product use. After balancing for baseline MLSI, beverage and tobacco use, fifty-four healthy adults with existing stain were randomly distributed into two comparable groups: Arm and Hammer Truly Radiant Rejuvenating toothpaste or Colgate Cavity Protection toothpaste (negative control). Within-treatment comparisons between baseline and day five were made using matched-pair t-tests, and between-treatment comparisons of MSLI scores were performed using ANCOVA, with baseline scores as covariates. Twenty-eight subjects in the Truly Radiant Rejuvenating toothpaste group and twenty-six subjects in the negative control group completed the study. The groups had comparable mean scores at baseline (p > 0.05). The Truly Radiant Rejuvenating toothpaste produced a statistically significant 23.1% total (composite) stain reduction from baseline after five days of product use (p 0.05). Between-treatment analysis showed statistically significantly (p toothpaste compared to the Colgate control following five days of product use. There were no adverse events reported during the study. The A&H Truly Radiant Rejuvenating toothpaste is safe and effective in reducing extrinsic stain compared to a regular toothpaste control.

  17. Lectin binding patterns and immunohistochemical antigen detection ...

    African Journals Online (AJOL)

    Ibrahim Eldaghayes

    2018-02-09

    Feb 9, 2018 ... the effect of infection with Brucella abortus on the pattern of lectin binding in bovine fetal lungs (n=6) and bovine placentas (n=5). Fetal lungs and placenta from heifers experimentally inoculated with B. abortus, strain 2308 were examined by histological, lectin-histochemical, immunohistochemical and ...

  18. Immunohistochemical evaluation of iron accumulation in term ...

    African Journals Online (AJOL)

    Classical immunohistochemical studies on placenta have shown that there is a linear increase in iron storage in the placenta in the first half of a normal pregnancy, however, these stocks are decreased in normal 3rd trimester placenta. Iron accumulation in term placentas of preeclamptic and normal pregnancies were ...

  19. A morphological and immunohistochemical evaluation of gastric ...

    African Journals Online (AJOL)

    Background: Gastric resections for carcinoma are common, but gastric carcinoma in South Africa, and particularly within the Western Cape province, has not been well documented. Method: The objective of the study was to immunohistochemically evaluate HER2/ neu overexpression, determine aberrations in β-catenin and ...

  20. The expression of MDM2 in gastrointestinal stromal tumors: immunohistochemical analysis of 35 cases.

    Science.gov (United States)

    Efared, Boubacar; Atsame-Ebang, Gabrielle; Tahiri, Layla; Sidibé, Ibrahim Sory; Erregad, Fatimazahra; Hammas, Nawal; Arifi, Samia; Mellouki, Ihsane; Ousadden, Abdelmalek; Mazaz, Khalid; El Fatemi, Hinde; Chbani, Laila

    2018-01-01

    Gastrointestinal stromal tumors (GIST) are the most common primary mesenchymal tumors of the digestive system. The assessment of their biological behavior still remains a scientific challenge. To date, there are no well-established biological prognostic markers of GIST. Our aim is to study the expression of the MDM2 oncoprotein in GIST through an immunohistochemical analysis. It was a retrospective study of 35 cases of GIST diagnosed from 2009 to 2012 in the department of pathology of Hassan II university hospital, Fès, Morocco. MDM2 immunohistochemical staining was performed on archival paraffin-embedded and formalin-fixed specimens (with a threshold of nuclear positivity > 10%). Analysis of correlations between MDM2 immunoexpression and clinicopathological features of GIST has been performed. The mean age was 55.23 years (range 25-84 years) with a male predominance (sex ratio = 1.5). The stomach was the main site of GIST, with 17 cases (48.57%) followed by the small bowel (9 cases, 25.71%). The spindle cell type GIST was the most frequent morphological variant (29 cases, 82.85%). Tumor necrosis was present in 8 cases (22.85%). Two patients (5.71%) had very low risk GIST, 5 (14.28%) had low risk GIST, 7 patients (20%) had intermediate risk tumors. The remaining 21 cases (60%) had high risk GIST. At the time of diagnosis, 9 patients (25.71%) had metastatic tumors. At immunohistochemical analysis, 40% of cases (14 patients) stained positive for MDM2. Of these MDMD2-positive tumors, 11/14 (78.57%) had high risk tumors and 8/14 cases (57.14%) presented with metastatic GIST. MDM2 positivity was significantly associated with the metastatic status ( p  = 0.001). The current study suggests that MDM2 immunohistochemical expression is a negative histoprognostic factor in GIST with a statistically significant correlation with metastasis.

  1. Histological stain evaluation for machine learning applications

    Directory of Open Access Journals (Sweden)

    Jimmy C Azar

    2013-01-01

    Full Text Available Aims: A methodology for quantitative comparison of histological stains based on their classification and clustering performance, which may facilitate the choice of histological stains for automatic pattern and image analysis. Background: Machine learning and image analysis are becoming increasingly important in pathology applications for automatic analysis of histological tissue samples. Pathologists rely on multiple, contrasting stains to analyze tissue samples, but histological stains are developed for visual analysis and are not always ideal for automatic analysis. Materials and Methods: Thirteen different histological stains were used to stain adjacent prostate tissue sections from radical prostatectomies. We evaluate the stains for both supervised and unsupervised classification of stain/tissue combinations. For supervised classification we measure the error rate of nonlinear support vector machines, and for unsupervised classification we use the Rand index and the F-measure to assess the clustering results of a Gaussian mixture model based on expectation-maximization. Finally, we investigate class separability measures based on scatter criteria. Results: A methodology for quantitative evaluation of histological stains in terms of their classification and clustering efficacy that aims at improving segmentation and color decomposition. We demonstrate that for a specific tissue type, certain stains perform consistently better than others according to objective error criteria. Conclusions: The choice of histological stain for automatic analysis must be based on its classification and clustering performance, which are indicators of the performance of automatic segmentation of tissue into morphological components, which in turn may be the basis for diagnosis.

  2. Cytogenetic and immunohistochemical characterization of mammary analogue secretory carcinoma of salivary glands.

    Science.gov (United States)

    Khurram, Syed A; Sultan-Khan, Jemel; Atkey, Neil; Speight, Paul M

    2016-12-01

    Mammary analogue secretory carcinoma (MASC), initially considered a subset of acinic cell carcinoma (ACC), harbors an ETV6 translocation [t(12:15)(p13:25 q)] and is now regarded as a distinct entity. Several putative markers to differentiate MASC from ACC have been reported; however, the immunohistochemical profile is still being explored and updated. The purpose of this study was to further explore the cytogenetic and immunohistochemical profile of MASC. Cases were analyzed for ETV6 translocation using fluorescent in situ hybridization and stained for CK8, amylase, mammaglobin, GCDFP-15, MUC1, MUC4, STAT5a, Ki-67 (n = 37), CK7, Cam5.2, CK14, SMA, p63, S100, vimentin and DOG1 (n = 42). Histochemical stains for mucins were also performed and data collected for age, sex, and site. Fluorescent in situ hybridization showed 9 cases with ETV6 rearrangement and 2 with increased ETV6 copies. These 11 cases showed an absence of PAS-D-resistant granules, with 10 of 11 showing strong S100, mammaglobin, and STAT5a staining. All ACCs showed diffuse DOG1 staining, whereas 8/11 MASCs were negative and 3 showed only focal DOG1 staining. DOG1 can be used in conjunction with PAS-D, S100, and mammaglobin to identify MASCs. Cases with increased ETV6 copies are a novel finding with a similar immunostaining profile and should be considered as MASCs. Copyright © 2016 Elsevier Inc. All rights reserved.

  3. Immunohistochemical study of pathological alterations of peritoneum in patients with terminal renal insufficiency and on peritoneal dialysis

    Directory of Open Access Journals (Sweden)

    Trbojević-Stanković Jasna

    2011-01-01

    Full Text Available Background/Aim. During peritoneal dialysis (PD an exchange of substances between blood and dialysate takes place through specific histological structures of peritoneum. Peritoneal double-layered serous membrane has, so far, mostly been studied with electron microscopy on experimental animals. The aim of this study was to assess integrity of peritoneal tissue in end-stage renal disease (ESRD and PD patients using standard light microscopy and immunohistochemical methods. Methods. Peritoneal tissue biopsies were performed on 25 persons: 8 healthy donors during nephrectomy, 9 ESRD patients upon insertion of PD catheter, and 8 PD patients upon removal of the catheter for medical indications. The samples were fixed and prepared routinely for immunocytochemical staining by standardized streptavidin biotin AEC method using a LSAB2® HRP kit (Dako®, Denmark for collagen IV and analyzed by light microscopy. Results. We observed mesothelial detachment from lamina propria, duplicated basement membrane and much thicker blood vessel walls in ESRD and PD patients, compared to healthy subjects. Differences in histological structure, emphasized with immunostaining, indicated pathological alterations of peritoneal tissue in the renal patients. Conclusions. Imunohistochemistry can be used in studying histological alterations of peritoneal tissue in ESRD and PD patients. This method may indicate possible problems in filtration and secretion processes in this tissue.

  4. Immunohistochemical analysis of the oxidative phosphorylation complexes in skeletal muscle from patients with mitochondrial DNA encoded tRNA gene defects.

    NARCIS (Netherlands)

    Paepe, B. De; Smet, J.; Lammens, M.M.Y.; Seneca, S.; Martin, J.J.; Bleecker, J. De; Meirleir, L. de; Lissens, W.; Coster, R. van

    2009-01-01

    BACKGROUND: Mitochondrial diseases display a heterogeneous spectrum of clinical phenotypes and therefore the identification of the underlying gene defect is often a difficult task. AIMS: To develop an immunohistochemical approach to stain skeletal muscle for the five multi-protein complexes that

  5. A critical appraisal of the immunohistochemical detection of the c-myc oncogene product in colorectal cancer.

    Science.gov (United States)

    Jones, D. J.; Ghosh, A. K.; Moore, M.; Schofield, P. F.

    1987-01-01

    Expression of c-myc was studied immunohistochemically in 100 colorectal carcinomas, using a monoclonal antibody, Myc 1-6E10, which is purported to recognize the oncoprotein (p62c-myc) in paraffin-embedded material. In normal epithelium, maturing crypt cells and terminally differentiated surface cells were positive, and proliferating basal crypt cells negative. All carcinomas stained positively, but intensity was independent of histological differentiation, Dukes' stage, DNA ploidy and survival. Staining was predominantly cytoplasmic despite the suspected nuclear location of p62c-myc and there was considerable staining of fibroblasts. When staining was compared in frozen and paraffin-embedded sections fixed in different ways, different patterns were observed. Acetone-fixed frozen sections exhibited weak nuclear and cytoplasmic staining or were negative. In formol-saline fixed frozen sections, there was stronger predominantly nuclear staining. In paraffin-embedded sections staining was predominantly cytoplasmic. This study suggests that c-myc expression is enhanced in the majority of colorectal carcinomas and although independent of clinical behaviour, may be a common event in malignant transformation. However, since staining is affected by fixation and processing, data obtained using Myc 1-6E10 on routinely processed specimens should be interpreted with caution. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 PMID:3325094

  6. Steroidogenic Factor 1, Pit-1, and Adrenocorticotropic Hormone: A Rational Starting Place for the Immunohistochemical Characterization of Pituitary Adenoma.

    Science.gov (United States)

    McDonald, William C; Banerji, Nilanjana; McDonald, Kelsey N; Ho, Bridget; Macias, Virgilia; Kajdacsy-Balla, Andre

    2017-01-01

    -Pituitary adenoma classification is complex, and diagnostic strategies vary greatly from laboratory to laboratory. No optimal diagnostic algorithm has been defined. -To develop a panel of immunohistochemical (IHC) stains that provides the optimal combination of cost, accuracy, and ease of use. -We examined 136 pituitary adenomas with stains of steroidogenic factor 1 (SF-1), Pit-1, anterior pituitary hormones, cytokeratin CAM5.2, and α subunit of human chorionic gonadotropin. Immunohistochemical staining was scored using the Allred system. Adenomas were assigned to a gold standard class based on IHC results and available clinical and serologic information. Correlation and cluster analyses were used to develop an algorithm for parsimoniously classifying adenomas. -The algorithm entailed a 1- or 2-step process: (1) a screening step consisting of IHC stains for SF-1, Pit-1, and adrenocorticotropic hormone; and (2) when screening IHC pattern and clinical history were not clearly gonadotrophic (SF-1 positive only), corticotrophic (adrenocorticotropic hormone positive only), or IHC null cell (negative-screening IHC), we subsequently used IHC for prolactin, growth hormone, thyroid-stimulating hormone, and cytokeratin CAM5.2. -Comparison between diagnoses generated by our algorithm and the gold standard diagnoses showed excellent agreement. When compared with a commonly used panel using 6 IHC for anterior pituitary hormones plus IHC for a low-molecular-weight cytokeratin in certain tumors, our algorithm uses approximately one-third fewer IHC stains and detects gonadotroph adenomas with greater sensitivity.

  7. Mucinous Cystadenoma of the Testis: A Case Report with Immunohistochemical Findings

    Directory of Open Access Journals (Sweden)

    Gilhyang Kim

    2017-03-01

    Full Text Available Mucinous cystadenoma of the testis is a very rare tumor. Herein, we report a case of mucinous cystadenoma arising in the testis of a 61-year-old man, along with a literature review. Computed tomography showed a 2.5-cm-sized poorly enhancing cystic mass. Grossly, the tumor was a unilocular cystic mass filled with mucinous material and confined to the testicular parenchyma. Histologically, the cyst had a fibrotic wall lined by mucinous columnar epithelium without atypia. Immunohistochemical staining was positive for cytokeratin 20 and CDX2, as well as focally positive for cytokeratin 7. The pathologic diagnosis was mucinous cystadenoma.

  8. Surgical management and immunohistochemical study of corneal plaques in vernal keratoconjunctivitis

    Directory of Open Access Journals (Sweden)

    Hsiu-Yi Lin

    2013-09-01

    Full Text Available Two children with shield ulcer in vernal keratoconjunctivitis unresponsive to steroid therapy received plaque removal by superficial keratectomy, followed by amniotic membrane transplantation (AMT. Hematoxylin and eosin staining of the excised corneal specimen revealed a thick layer of eosinophilic material attached to the Bowman's layer. These deposits were positive for eosinophil granule major basic protein, as confirmed by an immunohistochemical study. The shield ulcer healed after the amniotic membrane was removed. No recurrent corneal plaque developed, although corneal opacity complicated in both cases. Lamellar keratectomy with AMT offers an effective management by removing the cytotoxic plaques and protecting the denuded stroma from deposition of inflammatory debris.

  9. Tissue Staining (Chromoscopy of the Gastrointestinal Tract

    Directory of Open Access Journals (Sweden)

    M Brian Fennerty

    1999-01-01

    Full Text Available Tissue staining, or chomoscopy, is used as an adjunctive technique during gastrointestinal endoscopy. Chemical agents are applied to the gastrointestinal mucosal surface to identify specific epithelia or to enhance the mucosal surface characteristics of the gastrointestinal epithelium. This aids in the recognition of subtle lesions (ie, polyps or allows directed targeting of biopsies (ie, sprue or Barrett’s esophagus to increase the yield of endoscopic diagnostic accuracy. The four endoscopic tissue-staining techniques in use are vital staining, contrast staining (chromoscopy, reactive staining and tattooing. Some of the agents used for endoscopic tissue staining and the uses of chromoscopy in identifying pathology of the esophagus, stomach, small bowel and colon during endoscopy are discussed.

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

    Science.gov (United States)

    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

  11. Variations in immunohistochemical preservation of proteins in a mummification model.

    Science.gov (United States)

    Metcalfe, Ryan; Freemont, Tony

    2012-01-01

    Immunohistochemistry is an important tool in the investigation of ancient mummified remains because of its ability not only to detect proteins but also to isolate their location to specific tissues and thereby improve confidence that the results are genuine. A mouse model of Egyptian mummification has been used to demonstrate that the survival of proteins, judged by the retention of immunohistochemical staining, varies markedly. Some survive the process well, whereas others become barely detectable despite the morphology of the tissue being excellently preserved. The results obtained show that protein preservation is multi-factorial, with tissue type and degradation, and the properties of the protein itself all having significant effects. Proteins forming large, multi-subunit complexes such as collagen IV appear to be more resistant to degradation than those that do not, such as S-100. Although modern modelling studies cannot replicate the full extent of degradative processes and taphonomic changes experienced by real mummies, the results obtained can be useful for guiding research that requires ancient tissues. © 2011 The Authors. Journal of Anatomy © 2011 Anatomical Society of Great Britain and Ireland.

  12. Immunohistochemical evaluation of myofibroblast density in odontogenic cysts and tumors.

    Science.gov (United States)

    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 odontogenic tumors compared to odontogenic cysts (P 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.

  13. Pediatric meningiomas an aggressive subset: A clinicopathological and immunohistochemical study

    Directory of Open Access Journals (Sweden)

    M Hui

    2015-01-01

    Full Text Available Background: Meningiomas are uncommon neoplasms in the pediatric age group and differ in various aspects from their adult counterparts. They account for 0.4-4.6% of all primary brain tumors. Aims: To retrospectively analyze the clinicopathological and immunohistochemical features of pediatric meningiomas. Materials and Methods: Meningiomas in patients under 18 years of age diagnosed between January 2001 to December 2011 were analyzed retrospectively. The hematoxylin and eosin stained sections and Ki 67 labelling index (LI were reviewed for all the cases Results: The pediatric meningiomas accounted for 1.52% of total meningiomas (15/983. The mean age at presentation was 12 years with male to female ratio of 1.5:1. The presenting symptoms were headache, seizures, and motor deficits. The histology included 9 cases (60% of atypical meningioma (WHO grade II followed by 4 cases (26.67% of WHO grade-I and 2 cases (13.33% of anaplastic meningioma (WHO grade III. Five cases had a recurrence. Ki67 LI ranged from 0.5% to 1.5% in grade I, 0.5% to 15% in grade II and 13% to 24% in grade III meningiomas. Conclusion: Meningiomas are rare in children and show a male preponderance. There was a higher incidence of atypical and anaplastic meningiomas in pediatric population.

  14. Immunohistochemical evaluation of myofibroblast density in odontogenic cysts and tumors

    Science.gov (United States)

    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 cysts (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

  15. Clinicopathological and Immunohistochemical Characterisation of Gastric Schwannomas in 29 Cases

    Directory of Open Access Journals (Sweden)

    Lijun Zheng

    2014-01-01

    Full Text Available Schwannomas are tumors arising from the nervous system that also occur infrequently in the gastrointestinal tract, most commonly in the stomach. This report characterizes 29 patients with benign or malignant gastric schwannomas. Surgical data and clinical follow-up information were available for 28 cases with a median postoperative duration of 57 months. Clinicopathological and immunohistochemical characteristics of benign and malignant schwannomas were analysed. Four cases (13.7% were histologically diagnosed with malignant schwannoma. All tumors were positive for S-100 and CD56 proteins, displaying a diffuse staining pattern. Vimentin was expressed in 100% cases and all schwannomas were negative for smooth muscle actin, c-kit, and HMB-45. A significant difference was observed between the group of benign and malignant schwannomas as regards recurrences and metastasis after complete resection (P=0.015. The survival time of patients with benign schwannomas was longer than the malignant group (P=0.013, so gastric malignant schwannomas have a potential for recurrence and metastasis, with subsequently short survival. Complete resection with an attempt to remove all tumor tissue with negative margins is of paramount importance in the management of gastric schwannomas, particularly when they turn out to be malignant.

  16. Immunohistochemical diagnosis of fusariosis with monoclonal antibodies

    DEFF Research Database (Denmark)

    Jensen, H.E.; Aalbæk, B.; Jungersen, Gregers

    for fusariosis. A panel of newly developed Mabs for immunohistochemical diagnosis of fusariosis was screened for specificity on experimentally infected laboratory animal tissue and on skin tissue biopsies from two neutropenic patients with Fusarium sepsis. Methods: Somatic antigens were made from F. solani (CBS...... for establishing an accurate diagnosis. Although molecular techniques (e.g. in situ hybridization and PCR) have been explored for diagnostic use, the development of specific monoclonal antibodies (Mabs) for immunohistochemical identification of Fusarium spp. will extend the availability of diagnostic options...... containing homologous (fusariosis) and heterologous (aspergillosis, candidosis, and scedosporidiosis) fungal elements. Tissue reactive Mabs were then tested on skin biopsies from two patients with fusariosis sepsis with dissemination to the skin In the patients, a diagnosis of fusariosis-sepsis had been...

  17. Immunohistochemical prognostic indicators of lymphoma tumors

    African Journals Online (AJOL)

    STORAGESEVER

    2009-10-05

    Levine et al., 1991; Xerri et al.,. 1994; Kastan et al., 1995). The tumor suppressor .... antibody, then counter stained by haematoxylin dye). Specimen analysis. IHC evaluation was conducted by expert pathologist (Dr. Kanan F.,.

  18. A comparative analysis of angiopoietin 2 immunohistochemical ...

    African Journals Online (AJOL)

    2016-07-12

    , extension, and metastasis is not fully elucidated. The presented study aimed to investigate the relationship between Ang 2 staining intensity, expression rate in tumor tissue, and the stage of lung cancer. Materials and ...

  19. A comparative immunohistochemical study of aplastic thymuses for lymphocytic, epithelial, proliferative and apoptotic indices

    Directory of Open Access Journals (Sweden)

    Mahjoub F.

    2008-03-01

    Full Text Available Background: Immune deficiency is one of the major causes of morbidity and mortality in the modern world. Primary immunodeficiency comprises a wide range of disorders that mainly manifest in early childhood as devastating infections with opportunistic organisms. Thymic aplasia is found on autopsy of some patients afflicted with immune deficiency disorders, such as DiGeorge syndrome and severe combined immunodeficiency (SCID. After a thorough search of the literature, we found little information on the cellular characteristics of these thymuses. Our study aims to elucidate role of apoptosis in the pathogenesis of thymic aplasia and compare various lymphocytic and epithelial markers in normal and aplastic thymuses. Methods: We selected 12 subjects who died of severe infections with aplastic thymus found on autopsy, and 11 control subjects who died of unrelated causes, such as congenital heart disease. The presence of several markers, including Bcl2, P53, lymphocytic markers, and CD68, was examined using immunohistochemical methods on paraffin-embedded thymus sections. Positively-stained cells were counted per 1000 cells and the results stated as percentage of positive cells.                        Results: The mean age of the control group was between 7 days to 18 months (mean: 4.5 months. Parental consanguinity was present in 45.5% and 9.1% of the control and case groups, respectively; however, this was not statistically significant. We found significantly lower expression of Bcl2 in the case group (p value: 0.038. Furthermore, expression of CD68 was significantly higher in the case group. Epithelial markers were significantly higher in case subjects, although CD8 expression was higher in the control group. The presence of other markers was not significantly different between the two groups.Conclusions: Increase in apoptosis has a role in aplastic thymuses and prevention of apoptosis may halt this process. Also high CD68

  20. Immunohistochemical demonstration of glial markers in retinoblastomas

    DEFF Research Database (Denmark)

    Schrøder, H D

    1987-01-01

    Twenty retinoblastomas were studied immunohistochemically in order to visualize glial cells. In the retina, the glial cells in the ganglion cell layer and the Müller cells were GFAP positive, while only the glial cells of the ganglion cell layer expressed S-100 reactivity. In the tumours S-100/GF...... cells reactive for both S-100 and GFAP were demonstrated. The latter findings may represent differentiation in a glial direction in the more mature parts of retinoblastoma....

  1. Presence and possible significance of immunohistochemically demonstrable prolactin in breast apocrine metaplasia.

    Science.gov (United States)

    Kumar, S; Mansel, R E; Jasani, B

    1987-03-01

    Paraffin wax embedded formalin-fixed benign breast disease tissue taken from 17 patients (15 with microcystic disease and 2 with fibroadenoma) was studied for the presence of tissue bound prolactin using a rabbit antiserum against human prolactin applied in conjunction with a highly sensitive modified version of the dinitrophenyl (DNP)-hapten sandwich staining (DHSS) procedure. Sections taken from 14 of the 15 cases showing apocrine cystic changes exhibited strong prolactin staining restricted to the cytoplasm of metaplastic apocrine cells lining the cysts. Normal lobules and ducts and blunt duct proliferations were all negative, as were also the two cases of fibroadenoma. In contrast 6 out of 8 cases of breast cancer examined showed heterogenously distributed cytoplasmic staining in the cancer cells. Maximal prolactin staining in the apocrine cells was observed at antiserum dilutions as high as 1:60,000. This compared favourably with a 1:120,000 dilution that gave maximal levels of staining in the prolactotrophs present in serial sections taken from formalin fixed paraffin wax embedded post mortem human anterior pituitaries. In both types of tissues the specific staining was abolished by pre-absorption of the antiserum with human prolactin (10 micrograms ml-1). No staining was observed when the anti-prolactin serum was either omitted or substituted with DNP-labelled normal rabbit serum. Apocrine metaplasia in cystic disease of the breast has recently been found to be associated with an increased breast cancer risk. The strong and selective presence of immunohistochemically demonstrable prolactin in the metaplastic cells may be of significance in view of the hormone's known growth stimulating effect on the breast epithelium.

  2. Forehead wrinkles: a histological and immunohistochemical evaluation.

    Science.gov (United States)

    El-Domyati, Moetaz; Medhat, Walid; Abdel-Wahab, Hossam M; Moftah, Noha H; Nasif, Ghada A; Hosam, Wael

    2014-09-01

    Wrinkles are associated with cutaneous aging especially on sun-exposed skin. Despite they are considered a major topic in cosmetic dermatology, very few reports have studied the specific histological and immunohistochemical changes characteristic for wrinkles. The study aims to evaluate the histological and immunohistochemical changes of static forehead wrinkles in relation to surrounding photoaged skin. Biopsy specimens were obtained from the forehead wrinkles of 20 volunteers of Glogau's class III-IV wrinkles. Using histological and immunostaining methods coupled with computerized morphometric analysis, measurement of epidermal thickness and quantitative evaluation of total elastin and tropoelastin as well as collagen types I, III, and VII were performed for skin biopsies. In the wrinkle site, there was statistically significant lower epidermal thickness (P = 0.001), elastin (P wrinkle site and adjacent photoaged skin regarding collagen type I (P = 0.07) or III (P = 0.07). This study detected some histological and immunohistochemical differences in the wrinkle site when compared to adjacent photoaged skin. This may help in understanding the pathophysiology of facial wrinkling as well as its ideal way of management. © 2014 Wiley Periodicals, Inc.

  3. Immunohistochemical distribution of leptin in kidney tissues of melatonin treated diabetic rats.

    Science.gov (United States)

    Elis Yildiz, S; Deprem, T; Karadag Sari, E; Bingol, S A; Koral Tasci, S; Aslan, S; Nur, G; Sozmen, M

    2015-05-01

    We examined using immunohistochemistry the distribution of leptin in kidney tissues of melatonin treated, streptozotocin (STZ) diabetic rats. The animals were divided into five groups: control, sham, melatonin-treated, diabetic and melatonin-treated diabetic. Kidney sections were prepared and stained with hematoxylin and eosin, and Crossman's triple staining for histological examination. The immunohistochemical localization of leptin in the kidney tissue was determined using the streptavidin-biotin-peroxidase method. We determined that on days 7 and 14, the leptin immunoreactivity of the diabetic and melatonin-treated diabetic groups was weaker than for the other groups. Weak immunoreactivity was found in the proximal and distal tubules of the kidney in the diabetic and melatonin-treated diabetic groups on days 7 and 14, and strong immunoreactivity was found in the control, sham and melatonin groups. Melatonin application had no significant effect on leptin production in the kidney tissues of diabetic rats.

  4. Immunohistochemical analysis of collagen expression in human corpora lutea during the menstrual cycle and early pregnancy.

    Science.gov (United States)

    Iwahashi, Masaaki; Muragaki, Yasuteru; Ooshima, Akira; Umesaki, Naohiko

    2006-04-01

    To investigate the characteristic structure and function of human corpora lutea (CL), various types of collagen expression were determined in the CL tissues during the menstrual cycle and early pregnancy. In vitro experiment. Department of obstetrics and gynecology at a medical university. Regulatory cycling women and pregnant women with ovarian tumor and ectopic pregnancy who underwent adnexectomy. Immunohistochemistry for human type I, III, and IV collagen with specific monoclonal antibodies was used for analysis. Expression of type I, III, and IV collagen. Immunohistochemical staining for type I and III collagen revealed intense staining of the CL stroma during early pregnancy, as compared with those in the menstrual cycle. Moreover, pericellular intense immunostaining for type IV collagen was observed around the luteal cells, especially luteal granulosa cells, of early pregnancy. These results suggest that alterations in distribution of collagen might play an important role in determining the physiology and structure of the CL during the menstrual cycle and early pregnancy.

  5. Can SOX-10 or KBA.62 Replace S100 Protein in Immunohistochemical Evaluation of Sentinel Lymph Nodes for Metastatic Melanoma?

    Science.gov (United States)

    Vrotsos, Elena; Alexis, John

    2016-01-01

    Microscopic evaluation of sentinel lymph nodes for metastatic melanoma relies, in part, on the use of immunohistochemical analysis to identify minute metastatic deposits that may be overlooked on routine microscopy. At present S100 protein is widely used in this role, in large part for its superior sensitivity; however, interpretation is hampered by the presence of benign S100 protein-positive cellular elements present in every lymph node, leading to reduced specificity and consequent difficulties in interpretation. In recent years, multiple melanocytic markers have emerged that promise superior sensitivity and specificity, including KBA.62 and SOX-10. SOX-10 shows a nuclear pattern of staining. In normal tissue it is expressed in Schwann cells, melanocytes, and myoepithelial cells of salivary, bronchial, and mammary glands. KBA.62 is also specific except for staining of endothelial cells and shows a membranous staining pattern. This study was undertaken to determine whether KBA.62 or SOX-10 could equal (or surpass) the sensitivity of S100 protein while offering superior specificity in the immunohistochemical evaluation of sentinel lymph nodes for metastatic melanoma. In this study we performed immunohistochemical stains for S100 protein, Sox-10, and KBA.62 on 50 lymph nodes with proven metastatic melanoma. SOX-10 detected all cases of metastatic melanoma (50 of 50 cases; 100%) compared with S100 protein (48 of 50 cases; 96%) and KBA.62 (37 of 50 cases; 74%). There was no "background" staining of normal cellular elements with SOX-10 or KBA.62. In contrast, S100 protein was expressed in scattered dendritic interdigitating reticulum cells in the paracortex of lymph nodes, showing cytoplasmic and nuclear positivity, sometimes posing significant difficulty in differentiating benign reticulum cells from single cell metastatic melanoma. Our findings suggest that SOX-10 may be superior to S100 protein for identifying metastatic melanoma in a lymph node. KBA.62 was less

  6. Immunohistochemical evaluation of two corneal buttons with post-LASIK keratectasia.

    Science.gov (United States)

    Maguen, Ezra; Maguen, Barak; Regev, Lee; Ljubimov, Alexander V

    2007-09-01

    To examine immunohistochemically 2 human corneal buttons after corneal transplantation for post-laser in situ keratomileusis (LASIK) keratectasia. Two ectatic corneas after penetrating keratoplasty and 2 postmortem control corneas from a patient after uncomplicated LASIK were used. Cryostat sections were stained by immunofluorescence for >30 extracellular matrix (ECM) components and proteinases. The ratios of distance between LASIK flap interface and the upper epithelial layer to total corneal thickness were 0.27-0.34 in all cases. The whole flap interface was positive only for total and cellular fibronectin. Stromal types VI and XIV collagen, fibrillin-1, tenascin-C, and vitronectin were unchanged with no evidence of fibrosis. In ectasia cases, keratocytes adjacent to the flap did not express nidogens. Staining for type IV collagen alpha5 chain, nidogen-2, chains of laminin-8, and laminin-10 was weak and discontinuous in the epithelial basement membrane (EBM). Type IV collagen alpha1/alpha2 chains were found in the EBM of ectasia cases only. Matrix metalloproteinase (MMP)-10 showed increase in the epithelium, and MMP-3, in some keratocytes near the flap interface of ectatic corneas. Also, cathepsin F was seen at the flap margin only. Staining for limbal basal epithelial marker, alpha-enolase, was mostly absent in the ectatic cases, suggesting largely normal epithelial differentiation. Abnormal EBM structure similar to that previously observed in keratoconus and bullous keratopathy and an increase in certain proteinases suggest ongoing EBM lysis and remodeling. Immunohistochemical staining for fibronectin may be used to reveal the position of flap interface.

  7. Erbium doped stain etched porous silicon

    Energy Technology Data Exchange (ETDEWEB)

    Gonzalez-Diaz, B. [Departamento de Fisica Basica, Universidad de La Laguna, Avda. Astrofisico Francisco Sanchez, 38204 La Laguna, S/C de Tenerife (Spain); Diaz-Herrera, B. [Departamento de Energia Fotovoltaica, Instituto Tecnologico de Energias Renovables (ITER), Poligono Industrial de Granadilla, 38611 S/C Tenerife (Spain); Guerrero-Lemus, R. [Departamento de Fisica Basica, Universidad de La Laguna, Avda. Astrofisico Francisco Sanchez, 38204 La Laguna, S/C de Tenerife (Spain)], E-mail: rglemus@ull.es; Mendez-Ramos, J.; Rodriguez, V.D. [Departamento de Fisica Fundamental, Experimental Electronica y Sistemas, Universidad de La Laguna, Avda. Astrofisico Francisco Sanchez, 38204 La Laguna, S/C de Tenerife (Spain); Hernandez-Rodriguez, C. [Departamento de Fisica Basica, Universidad de La Laguna, Avda. Astrofisico Francisco Sanchez, 38204 La Laguna, S/C de Tenerife (Spain); Martinez-Duart, J.M. [Departamento de Fisica Aplicada, C-XII, Universidad Autonoma de Madrid, 28049 Cantoblanco, Madrid (Spain)

    2008-01-15

    In this work a simple erbium doping process applied to stain etched porous silicon layers (PSLs) is proposed. This doping process has been developed for application in porous silicon solar cells, where conventional erbium doping processes are not affordable because of the high processing cost and technical difficulties. The PSLs were formed by immersion in a HF/HNO{sub 3} solution to properly adjust the porosity and pore thickness to an optimal doping of the porous structure. After the formation of the porous structure, the PSLs were analyzed by means of nitrogen BET (Brunauer, Emmett and Teller) area measurements and scanning electron microscopy. Subsequently, the PSLs were immersed in a saturated erbium nitrate solution in order to cover the porous surface. Then, the samples were subjected to a thermal process to activate the Er{sup 3+} ions. Different temperatures and annealing times were used in this process. The photoluminescence of the PSLs was evaluated before and after the doping processes and the composition was analyzed by Fourier transform IR spectroscopy.

  8. Effect of toothpaste containing d-limonene on natural extrinsic smoking stain: a 4-week clinical trial.

    Science.gov (United States)

    Xie, Ping; Lu, Junjun; Wan, Huchun; Hao, Yuqing

    2010-08-01

    To determine whether natural smoking stain could be removed/inhibited effectively by a toothpaste containing 5% d-limonene. For comparison and contrast, the effects of d-limonene on tea stain were also assessed. The design was a randomized controlled double-blind trial with parallel groups. Toothpastes were: A: positive control with perlite whitening formulation; B: A+5% d-limonene; C: D + 5% d-limonene; D: negative control. The extrinsic stains were measured using Lobene Stain Index. Following baseline examination, all subjects were randomly assigned to one of the four toothpaste groups and instructed to brush with the assigned products twice daily. Subjects returned to the clinic after 4-week brushing for stain removal assessment, then all extrinsic stains, plaque and supragingival calculus were removed and use of assigned products was continued for another 4 weeks, and the stain scores were repeated for inhibition assessment. A total of 408 subjects, 201 with smoking stains and 207 with tea stains, participated in the trial. 5% d-limonene combined with Perlite whitening formulation significantly reduced stain scores both for smoking stain removal and inhibition (P 0.05). The additional advantage of 5% d-limonene was shown neither for removal nor for inhibition in the tea stain study (P > 0.05). All test products were well tolerated over the study period.

  9. Melanin Bleaching With Warm Hydrogen Peroxide and Integrated Immunohistochemical Analysis: An Automated Platform.

    Science.gov (United States)

    Liu, Chia-Hsing; Lin, Chih-Hung; Tsai, Min-Jan; Chen, Yu-Hsuan; Yang, Sheau-Fang; Tsai, Kun-Bow

    2018-02-01

    Diagnosing melanocytic lesions is among the most challenging problems in the practice of pathology. The difficulty of physically masking melanin pigment and the similarity of its color to commonly used chromogens often complicate examination of the cytomorphology and immunohistochemical staining results for tumor cells. Melanin bleach can be very helpful for histopathological diagnosis of heavily pigmented melanocytic lesions. Although various depigmentation methods have been reported, no standardized methods have been developed. This study developed a fully automated platform that incorporates hydrogen peroxide-based melanin depigmentation in an automated immunohistochemical analysis. The utility of the method was tested in 1 cell block of malignant melanoma cells in pleural effusion, 10 ocular melanoma tissue samples, and 10 cutaneous melanoma tissue samples. Our results demonstrated that the proposed method, which can be performed in only 3 hours, effectively preserves cell cytomorphology and immunoreactivity. The method is particularly effective for removing melanin pigment to facilitate histopathological examination of cytomorphology and for obtaining an unmasked tissue section for immunohistochemical analysis.

  10. Negative staining and cryo-negative staining of macromolecules and viruses for TEM.

    Science.gov (United States)

    De Carlo, Sacha; Harris, J Robin

    2011-02-01

    In this review we cover the technical background to negative staining of biomolecules and viruses, and then expand upon the different possibilities and limitations. Topics range from conventional air-dry negative staining of samples adsorbed to carbon support films, the variant termed the "negative staining-carbon film" technique and negative staining of samples spread across the holes of holey-carbon support films, to a consideration of dynamic/time-dependent negative staining. For each of these approaches examples of attainable data are given. The cryo-negative staining technique for the specimen preparation of frozen-hydrated/vitrified samples is also presented. A detailed protocol to successfully achieve cryo-negative staining with ammonium molybdate is given, as well as examples of data, which support the claim that cryo-negative staining provides a useful approach for the high-resolution study of macromolecular and viral structure. Copyright © 2009 Elsevier Ltd. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    M Devi

    2014-01-01

    Full Text Available 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.

  12. Identification of novel biomarker candidates for immunohistochemical diagnosis to distinguish low-grade chondrosarcoma from enchondroma.

    Science.gov (United States)

    Lai, Xianyin; Chen, Shaoxiong

    2015-07-01

    Chondrosarcoma is the third most common primary bone cancer, requiring surgical resection. However, differentiation of low-grade chondrosarcoma (grade 1) from enchondroma that is benign and only requires regular follow-up is one of the most frequent diagnostic dilemmas facing orthopedic oncologists in clinical management. Although multiple techniques are applied to make the distinction, immunohistochemistry is an important ancillary technique, especially when a histopathological stain of specimen must be obtained in order to guarantee an accurate confirmation. Currently, no adequate immunohistochemical diagnostic protein biomarkers are available to distinguish low-grade chondrosarcoma from enchondroma. To discover novel protein biomarker candidates, an LC-MS/MS approach was applied to directly compare formalin-fixed, paraffin-embedded low-grade chondrosarcoma with enchondroma tissue samples. The proteomics analysis revealed 17 protein biomarker candidates. A principle was developed to prioritize the candidates using category and ranking. An algorithm, prioritization index of biomarker candidates for immunohistochemistry on tissue specimens, was developed to rank the candidates inside each category. Using the proteomics data and bioinformatics results, the prioritization index of biomarker candidates for immunohistochemistry on tissue revealed periostin as a top candidate. Immunohistochemical staining of periostin in 23 low-grade chondrosarcoma and 31 enchondroma tissue specimens disclosed 87% specificity and 70% sensitivity. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Immunohistochemical features of multifocal melanoacanthoma in the hard palate: a case report

    Directory of Open Access Journals (Sweden)

    das Chagas e Silva de Carvalho Luis Felipe

    2013-01-01

    Full Text Available Abstract Background Melanoacanthoma (MA has been described in the oral mucosa as a solitary lesion or, occasionally, as multiple lesions. MA mainly affects dark skinned patients and grows rapidly, showing a plane or slightly raised appearance and a brown to black color. The differential diagnosis includes oral nevi, amalgam tattoos, and melanomas. We report here the case of a 58-year-old black woman who presented multiple pigmented lesions on the hard palate. Case presentation Based on the differential diagnosis of melanoma, a punch biopsy (4 mm in diameter was performed. The material was fixed in 10% formalin, embedded in paraffin, and stained with hematoxylin-eosin or submitted to immunohistochemical analysis. Immunohistochemistry using antibodies against protein S-100, melan-A, HMB-45, MCM-2, MCM-5, Ki-67 and geminin was performed. Immunohistochemical analysis revealed strong cytoplasmic immunoreactivity of dendritic melanocytes for proteinS-100, HMB-45 and melan-A.Positive staining for proliferative markers (MCM-2, MCM-5, Ki-67 was only observed in basal and suprabasal epithelial cells, confirming the reactive etiology of the lesion. The diagnosis was oral Melanoacanthoma (MA. Conclusion The patient has been followed up for 30 months and shows no clinical alterations. MA should be included in the differential diagnosis of pigmented lesions of the oral cavity.

  14. Immunohistochemical study of the expression of cell cycle regulating proteins at different stages of bladder cancer

    DEFF Research Database (Denmark)

    Primdahl, Hanne; von der Maase, Hans; Sørensen, Flemming Brandt

    2002-01-01

    PURPOSE: The cell cycle is known to be deregulated in cancer. We therefore analyzed the expression of the cell cycle related proteins p21, p27, p16, Rb, and L-myc by immunohistochemical staining of bladder tumors.METHODS: The tissue material consisted of bladder tumors from three groups of patients....... By immunohistochemical staining the protein expression was compared to allelic deletions of the corresponding genes. The allelic deletions were detected by PCR-based microsatellite analyses.RESULTS: We detected a significant reduction in the expression levels of the cell cycle related proteins p21(waf1) ( P=0.002), p27......(kip1) ( P=0.03), Rb ( P=0.00002), and L-myc ( P=0.00000007) in muscle invasive tumors compared to noninvasive tumors. Tumors presenting as muscle invasive at first diagnosis had significantly lower levels of p16/CDKN2A ( P=0.01) when compared to muscle invasive tumors that followed Ta or T1 precursor...

  15. Immunohistochemical study for the origin of ductular reaction in chronic liver disease.

    Science.gov (United States)

    Lee, Sun-Jae; Park, Jae-Bok; Kim, Kyung-Hyun; Lee, Woo-Ram; Kim, Jung-Yeon; An, Hyun-Jin; Park, Kwan-Kyu

    2014-01-01

    The appearance of proliferating bile ductular structures, which is called the "atypical ductular reaction" is frequently observed in various chronic liver diseases associated. However, the origin of these increased bile ductules has been a matter of controversy. In this study, we investigated the origin of ductular cells as an aspect of relation between epithelial to mesenchymal transition (EMT) and epithelial members of liver parenchyme, such as hepatocyte and cholangiocyte by immunohistochemical staining of human liver. Thirteen specimens of surgically resected liver with biliary cirrhosis were selected. Three sets of double immunohistochemical stains were done; Hep-Par 1 - cytokeratin 19 (CK19), Hep-Par 1 - α-sm ooth mus cle actin (α-SMA) and CK19 - α-SMA. As a result, we investigated the dual expression of the markers of hepatocyte and cholangiocyte in the same cell; in ductular cell and surrounding hepatocyte. However, there seems to be no dual expression of markers for EMT with epithelial markers. This study suggests a possibility of phenotypic change of mature hepatocyte into cholangiocyte. Future studies will be necessary to determine the role that proliferating cholangiocytes play in the pathogenesis of biliary fibrosis and how cholangiocytes interact with other cell types of the liver such as hepatic stellate cells or Kupffer cells.

  16. Adrenal medulla of AS/AGU rats: a histological and immunohistochemical study.

    Science.gov (United States)

    Al-Fayez, M A; Atteya, M; Mohamed, R A; Ahmed, A M; Alroalle, A H; Salah Khalil, M; Al-Ahmed, M; Payne, A

    2017-01-01

    The outcome of the autograft therapy for Parkinson's disease including autologous cells from adrenal medulla was disappointing. This could be attributed to the pathological process in Parkinson's disease affecting cells of the adrenal medulla. This study was performed to investigate the histopathological changes in the adrenal medulla of AS/AGU rat, a model of Parkinson's disease, in comparison with Albino Swiss (AS) rats. A total of 24 male AS rats were divided into four groups, each of 6 animals: AS W1 - AS rats aged 1 week; AS adult - AS adult rats; AS/ /AGU W1 - AS/AGU rats aged 1 week; and AS/AGU adult - AS/AGU adult rats. The rats were sacrificed and the adrenal glands were dissected and processed for histological staining with haematoxylin and eosin and periodic acid Schiff and for immunohistochemical staining for S100 protein, ubiquitin and tyrosine hydroxylase. The histological investigation of the adrenal medulla of AS/AGU rats showed vascular congestion, inflammatory cellular infiltration, pyknotic nuclei, necrotic chromaffin cells and medullary inclusion bodies. The immunohistochemical investigation of AS/AGU rats showed a statistically significant decrease in the expression of S100 protein, ubiquitin and tyrosine hydroxylase compared to AS rats. The histological and immunohistological changes in the adrenal medulla could explain the failure of outcome of adrenal autograft therapy in Parkinson's disease.

  17. Hyperspectral imaging of the crime scene for detection and identification of blood stains

    Science.gov (United States)

    Edelman, G. J.; van Leeuwen, T. G.; Aalders, M. C. G.

    2013-05-01

    Blood stains are an important source of information in forensic investigations. Extraction of DNA may lead to the identification of victims or suspects, while the blood stain pattern may reveal useful information for the reconstruction of a crime. Consequently, techniques for the detection and identification of blood stains are ideally non-destructive in order not to hamper both DNA and the blood stain pattern analysis. Currently, forensic investigators mainly detect and identify blood stains using chemical or optical methods, which are often either destructive or subject to human interpretation. We demonstrated the feasibility of hyperspectral imaging of the crime scene to detect and identify blood stains remotely. Blood stains outside the human body comprise the main chromophores oxy-hemoglobin, methemoglobin and hemichrome. Consequently, the reflectance spectra of blood stains are influenced by the composite of the optical properties of the individual chromophores and the substrate. Using the coefficient of determination between a non-linear least squares multi-component fit and the measured spectra blood stains were successfully distinguished from other substances visually resembling blood (e.g. ketchup, red wine and lip stick) with a sensitivity of 100 % and a specificity of 85 %. The practical applicability of this technique was demonstrated at a mock crime scene, where blood stains were successfully identified automatically.

  18. Bcl-2 expression is altered with ovarian tumor progression: an immunohistochemical evaluation

    Directory of Open Access Journals (Sweden)

    Anderson Nicole S

    2009-10-01

    Full Text Available Abstract Background Ovarian cancer is the most lethal gynecologic malignancy. The ovarian tumor microenvironment is comprised of tumor cells, surrounding stroma, and circulating lymphocytes, an important component of the immune response, in tumors. Previous reports have shown that the anti-apoptotic protein Bcl-2 is overexpressed in many solid neoplasms, including ovarian cancers, and contributes to neoplastic transformation and drug-resistant disease, resulting in poor clinical outcome. Likewise, studies indicate improved clinical outcome with increased presence of lymphocytes. Therefore, we sought to examine Bcl-2 expression in normal, benign, and cancerous ovarian tissues to determine the potential relationship between epithelial and stromal Bcl-2 expression in conjunction with the presence of lymphocytes for epithelial ovarian tumor progression. Methods Ovarian tissue sections were classified as normal (n = 2, benign (n = 17 or cancerous (n = 28 and immunohistochemically stained for Bcl-2. Bcl-2 expression was assessed according to cellular localization, extent, and intensity of staining. The number of lymphocyte nests as well as the number of lymphocytes within these nests was counted. Results While Bcl-2 staining remained cytoplasmic, both percent and intensity of epithelial and stromal Bcl-2 staining decreased with tumor progression. Further, the number of lymphocyte nests dramatically increased with tumor progression. Conclusion The data suggest alterations in Bcl-2 expression and lymphocyte infiltration correlate with epithelial ovarian cancer progression. Consequently, Bcl-2 expression and lymphocyte status may be important for prognostic outcome or useful targets for therapeutic intervention.

  19. Tympanomastoid cholesterol granulomas: Immunohistochemical evaluation of angiogenesis.

    Science.gov (United States)

    Iannella, Giannicola; Di Gioia, Cira; Carletti, Raffaella; Magliulo, Giuseppe

    2017-08-01

    This study investigates the immunohistochemical expression of vascular endothelial growth factor (VEGF) and CD34 in patients treated for middle ear and mastoid cholesterol granulomas to evaluate the angiogenesis and vascularization of this type of lesion. A correlation between the immunohistochemical data and the radiological and intraoperative evidence of temporal bone marrow invasion and blood source connection was performed to validate this hypothesis. Retrospective study. Immunohistochemical expression of VEGF and CD34 in a group of 16 patients surgically treated for cholesterol granuloma was examined. Middle ear cholesteatomas with normal middle ear mucosa and external auditory canal skin were used as the control groups. The radiological and intraoperative features of cholesterol granulomas were also examined. In endothelial cells, there was an increased expression of angiogenetic growth factor receptors in all the cholesterol granulomas in this study. The quantitative analysis of VEGF showed a mean value of 37.5, whereas the CD34 quantitative analysis gave a mean value of 6.8. Seven patients presented radiological or intraoperative evidence of bone marrow invasion, hematopoietic potentialities, or blood source connections that might support the bleeding theory. In all of these cases there was computed tomography or intraoperative evidence of bone erosion of the middle ear and/or temporal bone structures. The mean values of VEGF and CD34 were 41.1 and 7.7, respectively. High values of VEGF and CD34 are present in patients with cholesterol granulomas. Upregulation of VEGF and CD34 is indicative of a remarkable angiogenesis and a widespread vascular concentration in cholesterol granulomas. 3b. Laryngoscope, 127:E283-E290, 2017. © 2017 The American Laryngological, Rhinological and Otological Society, Inc.

  20. Phenotypic and immunohistochemical characterization of sarcoglycanopathies

    Directory of Open Access Journals (Sweden)

    Ana F. B. Ferreira

    2011-01-01

    Full Text Available INTRODUCTION: Limb-girdle muscular dystrophy presents with heterogeneous clinical and molecular features. The primary characteristic of this disorder is proximal muscular weakness with variable age of onset, speed of progression, and intensity of symptoms. Sarcoglycanopathies, which are a subgroup of the limb-girdle muscular dystrophies, are caused by mutations in sarcoglycan genes. Mutations in these genes cause secondary deficiencies in other proteins, due to the instability of the dystrophin-glycoprotein complex. Therefore, determining the etiology of a given sarcoglycanopathy requires costly and occasionally inaccessible molecular methods. OBJECTIVE: The aim of this study was to identify phenotypic differences among limb-girdle muscular dystrophy patients who were grouped according to the immunohistochemical phenotypes for the four sarcoglycans. METHODS: To identify phenotypic differences among patients with different types of sarcoglycanopathies, a questionnaire was used and the muscle strength and range of motion of nine joints in 45 patients recruited from the Department of Neurology - HC-FMUSP (Clinics Hospital of the Faculty of Medicine of the University of São Paulo were evaluated. The findings obtained from these analyses were compared with the results of the immunohistochemical findings. RESULTS: The patients were divided into the following groups based on the immunohistochemical findings: a-sarcoglycanopathies (16 patients, b-sarcoglycanopathies (1 patient, y-sarcoglycanopathies (5 patients, and nonsarcoglycanopathies (23 patients. The muscle strength analysis revealed significant differences for both upper and lower limb muscles, particularly the shoulder and hip muscles, as expected. No pattern of joint contractures was found among the four groups analyzed, even within the same family. However, a high frequency of tiptoe gait was observed in patients with a-sarcoglycanopathies, while calf pseudo-hypertrophy was most common in

  1. Immunohistochemical assessment of oestrogen and progesterone receptors

    DEFF Research Database (Denmark)

    Grabau, D A; Thorpe, S M; Knoop, A

    2000-01-01

    Two different methods to determine steroid receptors were analysed with respect to their ability to estimate prognosis in primary breast cancer patients. The immunohistochemical assay (IHA) was compared with the dextran-coated charcoal (DCC) method of receptor determination. A random sample of 281...... of patients, receptor positive cases fared better than negative cases in all situations. Investigation of the prognostic power revealed that classification based on IHA allowed better discrimination of patients than classification based on the DCC method. The reason for this difference might be because...

  2. Immunohistochemical Expressions of MUC2, MUC5AC, and MUC6 in Normal, Helicobacter pylori Infected and Metaplastic Gastric Mucosa of Children and Adolescents.

    Science.gov (United States)

    Park, Ji Sook; Yeom, Jung-Sook; Seo, Ji-Hyun; Lim, Jae-Young; Park, Chan-Hoo; Woo, Hyang-Ok; Youn, Hee-Shang; Jun, Jin-Su; Park, Ji-Hoe; Ko, Gyung-Hyuck; Baik, Seung-Chul; Lee, Woo-Kon; Cho, Myung-Je; Rhee, Kwang-Ho

    2015-08-01

    The aim of this study was to investigate expression of gastric mucins in children and adolescents and to assess their relations with age and Helicobacter pylori (H. pylori) infection. Gastric biopsies were collected from 259 pediatric and adulthood patients with gastrointestinal symptoms among all of patients undergone gastroduodenoscopy from 1990 to 2004 at Gyeongsang National University hospital and assorted based on H. pylori infection, age, and intestinal metaplasia as follows; H. pylori infection before 5 years of age or not, H. pylori infection between 5 and 9 years of age or not, H. pylori infection between 10 and 14 years of age or not, H. pylori infection between 20 and 29 years of age or not and intestinal metaplasia between 21 and 35 years of age. Total 810 tissue slides from the subjects were examined regarding expressions of Mucin2 (MUC2), Mucin5AC (MUC5AC), and Mucin6 (MUC6) in nine groups using immunohistochemical stains. A semiquantitative approach was used to score the staining extent of tissue slide. Increased expressions of MUC2, MUC5AC, and MUC6 were noted in intestinal metaplasia compared with subjects infected with H. pylori between 20 and 29 years. Gastric expressions of MUC5AC were decreased in older than 5 years with H. pylori compared with in older than 5 years without H. pylori (p pylori status. Some nuclear expressions of MUC2 and MUC6 were noted in children without intestinal metaplasia. MUC5AC might be affected by chronic H. pylori infection. In addition to biomarkers for intestinal metaplasia or prognostic factors for gastric cancer in adults, MUC2 and MUC6 in children might have an another role, based on ectopic gastric nuclear expressions of MUC2 and MUC6 in children without intestinal metaplasia. © 2015 John Wiley & Sons Ltd.

  3. A Randomized Controlled Clinical Trial to Evaluate Extrinsic Stain Removal of a Whitening Dentifrice.

    Science.gov (United States)

    Terézhalmy, Géza; He, Tao; Anastasia, Mary Kay; Eusebio, Rachelle

    2016-12-01

    To evaluate the extrinsic stain removal efficacy of a new whitening dentifrice containing sodium hexametaphosphate (SHMP) over a two-week period. This study used a controlled and randomized, examiner-blind, single-center, two-treatment, parallel group design. Subjects with visible extrinsic dental stain on facial surfaces of their anterior teeth, and meeting all study criteria, were entered into the trial. The test group received the whitening dentifrice with sodium fluoride and SHMP and an ADA reference soft manual toothbrush. Subjects in the control group received a dental prophylaxis after the initial examination at Baseline and were instructed to use their usual oral hygiene products at home. Subjects returned at Day 3 and Week 2 for re-evaluation of extrinsic dental stain. Extrinsic stain was measured using the Interproximal Modified Lobene (IML) Stain Index; safety was assessed based on clinical examination. Fifty subjects (mean age 32.0 years) completed the study, with 25 in each group. Statistically significant reductions in composite stain for whole tooth, as well as interproximal, gingival, and body surfaces were observed for both groups at Day 3 and Week 2 (p 0.3). At Day 3, median percent reductions in composite IML stain from Baseline were 98% for the prophylaxis group and 100% for the test dentifrice group. At Week 2, median percent reductions in composite IML stain were 100% compared to Baseline for both groups. No adverse events were reported for either group. The whitening dentifrice demonstrated a statistically significant reduction in IML stain after three days and two weeks of use relative to baseline. Stain reduction with the toothpaste was comparable to a dental prophylaxis.

  4. Crossover clinical investigation of a whitening chewing gum for inhibiting dental stain formation in conjunction with tooth brushing.

    Science.gov (United States)

    Milleman, Jeffery L; Milleman, Kimberly R; Kleber, Carl J; Proskin, Howard M; Dodds, Michael; Kelley, Michael; Ramirez, Lilian

    2014-01-01

    The purpose of this clinical investigation was to evaluate the effectiveness of a marketed whitening chewing gum compared to a no-gum control in preventing the formation of extrinsic stains on the teeth of stain-forming subjects when chewed over a 12-week period of regular unsupervised use in conjunction with daily tooth brushing. This was a single-center, examiner-blind, randomized, 12-week crossover clinical trial. Stain-forming (after smoking or drinking coffee or tea) adults, starting with a stain-free baseline, either chewed the test gum (Orbit White) unsupervised four times per day, 15 minutes/chew, or used no gum along with daily brushing with a commercially available toothbrush and dentifrice for 12 weeks. At the crossover, all procedures were repeated with subjects assigned the opposite treatment. Extrinsic stain was measured at six and 12 weeks by both the Lobene Stain Index (LSI) and the Modified Lobene Stain Index (MLSI) using separate experienced examiners. After 12 weeks, LSI stain scores showed a significant 25% reduction (p = 0.0008) in new stain formation for subjects using the test chewing gum along with tooth brushing versus tooth brushing alone (no-gum control). The corresponding MLSI stain scores demonstrated a 36% reduction (p teeth. The overall findings of this clinical study demonstrated that regular use of Orbit White chewing gum, soon after smoking or drinking coffee or tea, will supplement daily tooth brushing in preventing unsightly stains from forming on the anterior teeth compared to brushing alone.

  5. Utility of 5-Methylcytosine Immunohistochemical Staining to Assess Global DNA Methylation and Its Prognostic Impact in MDS Patients

    Science.gov (United States)

    Chandra, Dinesh; Tyagi, Seema; Singh, Jasdeep; Deka, Roopam; Manivannan, Prabhu; Mishra, Pravas; Pati, Hara Prasad; Saxena, Renu

    2017-12-29

    Background: DNA methylation plays a vital role in the pathogenesis of the myelodysplastic syndrome (MDS), a heterogeneous group of clonal hematopoietic stem cell (HSC) disorders. It is reported to be an independent prognostic factor affecting overall survival (OS). Our aim was to analyze the role of global DNA methylation using an anti-5-methylcytosine (5-MC) antibody by immunohistochemistry (IHC) of bone marrow biopsy (BM Bx) specimens in MDS patients, assessing correlations with various clinical and biological prognostic factors. Material and methods: A total of 59 MDS cases, classified as per the World Health Organization (WHO) 2008 guidelines, were evaluated over a period of 4 years. Clinical data were retrieved from departmental case records and anti-5-MC expression was analyzed with formalin fixed paraffin embedded sections of BM Bx specimens of MDS patients and controls. Results: The median age at diagnosis was 52 years (15-85years). Patients were categorized into low risk (59%) and high risk (41%) according to International Prognostic Scoring System (IPSS). The median follow-up time was 10 months (1 to 37 months). We generated a methylation score (M-score) using anti-5-MC and with the derived cut-off of 30.5 from the receiver operator curve (ROC), there was a significant difference between the two groups in the percentage of BM blasts (p=0.01), WHO sub-type (p=0.01), IPSS (p=0.004), progression to AML (p=0.04) on univariate analysis. Interestingly, patients showing a high M-score (M-score ≥ 30.5) demonstrated a significantly shorter OS and progression to AML. However, on multivariate analysis, only BM blasts (p=0.01) and IPSS (p=0.02) remained independent variables for progression to AML and OS respectively. Conclusion: Immunostaining with anti-5-MC antibody with BM Bx samples is a simple and cost effective technique to detect global methylation, a powerful tool to predict overall survival in patients with MDS. Creative Commons Attribution License

  6. [Value of immunohistochemical staining with mutation-specific antibodies in detecting EGFR mutations: a meta-analysis].

    Science.gov (United States)

    Ma, Qing; Wang, Jing; Zhong, Diansheng; Ning, Chao; Liu, Chang; Xiao, Ping

    2014-06-20

    It has been proven that epidermal growth factor receptor (EGFR) mutation is the most important predictive factor for determining the effect of EGFR tyrosine kinase inhibitors (TKIs) applied to non-small cell lung cancer (NSCLC) patients. The patients with EGFR mutations response better to TKIs. To detect EGFR mutation has been particularly essential to select first-line treatment for lung cancer patients. To research and analyze the sensitivity and specificity of immunohistochemistry (IHC) using mutation specific antibodies in detecting EGFR mutations compared with DNA sequencing, and further evaluate the accuracy and clinical application value of IHC. All required articles in Pubmed database were searched. The deadline of retrieval was March 26, 2013. Then further screening the articles based on the inclusion and exclusion criteria. Meta analysis of diagnostic test was applied to analyze the sensitivity and specificity of IHC compared with DNA sequencing for the detection of EGFR mutations. Ten articles were included in the meta analysis, there were 1,679 samples in L858R group and 1,041 samples in E746-A750del group. The DOR were 225.17 (95%CI: 55.67-910.69) and 267.16 (95%CI: 132.45-538.88) respectively; the AUC of SROC were 0.948,4 (SEAUC=0.014,4) and 0.981,3 (SEAUC=0.009,9) respectively; the Q values were 0.888,3 (SEQ*=0.019,2) and 0.939,7 (SEQ*=0.019,1) respectively. The specificity and sensitivity of IHC method using these two mutation-specific antibodies were relatively high. As a screening method for EGFR mutations, the IHC with mutation specific antibodies is of clinical value.

  7. [Comparison of collagen fiber staining between Van-Gieson staining and Masson trichrome staining of hepatic specimens in mice with Schistosoma japonicum infection].

    Science.gov (United States)

    Huang, Da-Ke; Zhang, Yu-Xia; Man, Su-Qin; Yu, Fa-Zhi; Shen, Ji-Jia

    2012-08-01

    To compare the effects of collagen fiber staining between Van-Gieson staining and Masson trichrome staining of hepatic specimens in mice with Schistosoma japonicum infection. A model of hepatic granuloma and fibrosis was established by infecting mice with S. japonicum cercariae, then the hepatic specimens were taken and Van-Gieson staining and Masson trichrome staining were performed. Eventually, the area of granuloma and fibrosis were measured by imaging analysis software. When the time of staining was 3-7 min, there was no significant difference of the fibrosis areas between the two methods (P > 0.05); when the time of staining was more than 10 min, the staining area showed by Masson's staining was significantly larger than that showed by Van-Gieson staining, and the difference was statistically significant (P Masson trichrome staining, therefore Van-Gieson staining is a better method to display collagen.

  8. Immunohistochemical visualization of mouse interneuron subtypes

    DEFF Research Database (Denmark)

    Jensen, Simon Mølgaard; Ulrichsen, Maj; Boggild, Simon

    2014-01-01

    on the market directed against these four markers. Searches in the literature databases allowed us to narrow it down to a subset of antibodies most commonly used in publications. However, in our hands the most cited ones did not work for immunofluorescence stainings of formaldehyde fixed tissue sections...

  9. Immunohistochemical Assessment of Proliferating Cell Nuclear ...

    African Journals Online (AJOL)

    Background: Proliferating cell nuclear antigen (PCNA) is a nuclear protein synthesized in the late G1 and S‑phase of the cell cycle. ... Two sections were taken from each one for H and E. Other sections were stained according to super sensitive polymer horseradish peroxidase method for identifying PCNA expression.

  10. Intracellular and juxtacellular staining with biocytin.

    Science.gov (United States)

    Wilson, Charles J; Sachdev, R N S

    2004-05-01

    Many physiological studies require microscopic examination of the recorded neuron for identification. This unit describes how intracellular and extracellular recording can be combined with single-neuron staining to enable sequential physiological and morphological studies.

  11. Homogeneous luminescent stain etched porous silicon elaborated by a new multi-step stain etching method

    Energy Technology Data Exchange (ETDEWEB)

    Hajji, M., E-mail: mhajji2001@yahoo.fr [Laboratoire de Photovoltaïque, Centre de Recherche et des Technologies de l’Energie, Technopôle de Borj-Cédria BP 95, Hammam-Lif 2050 (Tunisia); Institut Supérieur d’Electronique et de Communication de Sfax, route Menzel Chaker Km 0.5, BP 868, Sfax 3018 (Tunisia); Khalifa, M.; Slama, S. Ben; Ezzaouia, H. [Laboratoire de Photovoltaïque, Centre de Recherche et des Technologies de l’Energie, Technopôle de Borj-Cédria BP 95, Hammam-Lif 2050 (Tunisia)

    2013-11-01

    This paper presents a new method to produce porous silicon which derived from the conventional stain etching (SE) method. But instead of one etching step that leads to formation of porous layer, the substrate is subjected to an initial etching step with a duration Δt{sub 0} followed by a number of supplementary short steps that differs from a layer to another. The duration of the initial step is just the necessary time to have a homogenous porous layer on the whole surface of the substrate. It was found that this duration is largely dependent of the doping type and level of the silicon substrate. The duration of supplementary steps was kept as short as possible to prevent the formation of bubbles on the silicon surface during silicon dissolution which leads generally to inhomogeneous porous layers. It is found from surface investigation by atomic force microscopy (AFM) that multistep stain etching (MS-SE) method allows to produce homogeneous porous silicon nanostructures compared to the conventional SE method. The chemical composition of the obtained porous layers has been evaluated using Fourier transform infrared spectroscopy (FTIR). Photoluminescence (PL) measurement shows that porous layers produced by SE and MS-SE methods have comparable spectra indicating that those layers are composed of nanocrystallites with comparable sizes. But the intensity of photoluminescence of layer elaborated by MS-SE method is higher than that elaborated by the SE method. Total reflectance characteristics show that the presented method allows the production of porous silicon layers with controllable thicknesses and optical properties. Results for porous silicon layers elaborated on heavily doped n-type silicon show that the reflectance can be reduced to values less than 3% in the major part of the spectrum.

  12. Immunohistochemical Detection of Cancer Stem Cell Related Markers CD44 and CD133 in Metastatic Colorectal Cancer Patients

    Directory of Open Access Journals (Sweden)

    Pavel Pitule

    2014-01-01

    Full Text Available Aim. The goal of this study was to semiquantitatively detect presence of cancer stem cells markers CD44 and CD133 in immunohistochemically stained paired samples of colorectal cancer (CRC and colorectal liver metastases (CLM. Level of staining intensity was compared to clinical and pathological characteristics of tumors with the aim to identify impact of CD44 or CD133 expression on tumor behavior. Patients and Methods. Formalin fixed paraffin embedded samples from 94 patients with colorectal tumor and liver metastases were collected at Sikl’s Department of Pathology. Samples were stained by antibodies against CD44 and CD133. Presence and intensity of staining was assessed semiquantitatively by three trained researchers. Results. Patients with higher level of CD133 staining in CRC had longer disease free interval (Cox-Mantel P=0.0244, whereas we found no relation between CD44 expression and overall survival or disease free interval. CD133 expression in CRC and CLM differed based on CRC grading; in case of CD44 we found differences in staining intensity in individual stages of tumor lymph node invasion. Conclusion. Effect of cancer stem cell markers on prognosis of colorectal cancer can vary depending on pathological classification of tumor, and we have shown that CD133, generally considered to be a negative marker, can bear also clinically positive prognostic information in group of patients with colorectal liver metastases.

  13. Collagenous gastritis: a morphologic and immunohistochemical study of 40 patients.

    Science.gov (United States)

    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.

  14. Dural invasion of meningioma: a histological and immunohistochemical study.

    Science.gov (United States)

    Nagashima, Goro; Fujimoto, Tsukasa; Suzuki, Ryuta; Asai, Jun-ichiro; Itokawa, Hiroshi; Noda, Masayuki

    2006-04-01

    Meningioma usually grows and expands into the brain, but invasion into the brain parenchyma is relatively rare. Meningioma arises from arachnoid cap cells, and infiltration into dura mater is the main growth pattern of meningiomas. However, little is known about the mechanism of meningioma invasion into the dura mater. In this study, seven specimens, including dural attachments, from seven cases of meningioma were used for immunohistochemical analysis. Matrix metalloproteinase (MMP)-1, -2, -9, urokinase-type plasminogen activator (uPA), vascular endothelial growth factors (VEGF), flt-1, E-cadherin, estrogen receptor (EgR), progesterone receptor (PgR), and aquaporin (AQP)-1, -4 were used as primary antibodies. There were several patterns of meningioma invasion into the dura mater: papillary-shaped invasion with destruction of dural structure, infiltration along the fibers of the dura mater, and invasion of several tumor cell units with fibroblast infiltration. Strong immunostaining was obtained with MMP-1, followed by AQP-1 and uPA, within the invading tumor cells. Neovasculature and extravasated erythrocytes, which stained with AQP-1, were also occasionally observed around the invading tumor cells. Simpson grade II removal of meningiomas results in high recurrence rates, and the inhibition of meningioma growth via dural invasion will facilitate improved remission in many cases with meningioma. In this study, MMP-1, AQP-1, and uPA are considered to have some role in the dural infiltration of meningioma cells. The fact that AQP-1 was highly expressed at the dural attachment and invading front of meningioma may indicate that dural invasion of the meningioma may be facilitated by AQP-1-induced water flow and neovascularization.

  15. Clinicopathological and immunohistochemical features of primary central nervous system germ cell tumors: a 24-years experience.

    Science.gov (United States)

    Gao, Yuping; Jiang, Jiyao; Liu, Qiang

    2014-01-01

    Primary central nervous system (CNS) germ cell tumors (GCTs) are a rare heterogeneous group of lesions, which the clinicopathological features have a marked degree of heterogeneity comparing with that of gonadal GCTs. Accurately diagnosing CNS GCTs might be extremely difficult and requires immunohistochemical verification. This study was to investigate the biological feature of CNS GCTs and diagnostic value of immunohistochemical markers OCT3/4, C-kit, PLAP, and CD30 in CNS GCTs. A retrospective study was performed on 34 patients with CNS germ cell tumors between 1990 and 2014. 34 CNS GCTs account for 9.2% of all primary CNS neoplasms. The sellar region (35.3%) and pineal gland (17.6%) were the most common sites of intracranial GCTs. Hydrocephalus (82.4%) and diplopia (46.9%) were the two most common clinical presentations. The most common histological subtypes were germinoma (67.6%). PLAP, c-kit, OCT3/4 were highly expressed in gernimomas. CD30 and CK AE1/3 stainings were positive in embryonal carcinoma. Yolk sac tumor component showed positive staining for AFP and CK AE1/3. β-HCG staining was positive in choriocarcinoma and STGC. Patients with mature teratomas and germinomas had a better prognosis (a 5-year survival rate) than those with embryonal carcinoma and choriocarcinoma (a 5-year survival rates were 0). Our finding suggest that the incidences of primary CNS GCTs are higher in South China than in the West, but mixed GCTs are uncommon in our study. The judicious use of a panel of selected markers is helpful in diagnosing and predicting the prognosis for CNS GCTs.

  16. A simple protocol for paraffin-embedded myelin sheath staining with osmium tetroxide for light microscope observation.

    Science.gov (United States)

    Di Scipio, Federica; Raimondo, Stefania; Tos, Pierluigi; Geuna, Stefano

    2008-07-01

    Experimental investigation of peripheral nerve fiber regeneration is attracting more and more attention among both basic and clinical researchers. Assessment of myelinated nerve fiber morphology is a pillar of peripheral nerve regeneration research. The gold standard for light microscopic imaging of myelinated nerve fibers is toluidine blue staining of resin-embedded semithin sections. However, many researchers are unaware that the dark staining of myelin sheaths typically produced by this procedure is due to osmium tetroxide postfixation and not due to toluidine blue. In this article, we describe a simple pre-embedding protocol for staining myelin sheaths in paraffin-embedded nerve specimens using osmium tetroxide. The method involves immersing the specimen in 2% osmium tetroxide for 2 h after paraformaldehyde fixation, followed by routine dehydration and paraffin embedding. Sections can then be observed directly under the microscope or counterstained using routine histological methods. Particularly good results were obtained with Masson's trichrome counterstain, which permits the imaging of connective structures in nerves that are not detectable in toluidine blue-stained resin sections. Finally, we describe a simple protocol for osmium etching of sections, which makes further immunohistochemical analysis possible on the same specimens. Taken together, our results suggest that the protocol described in this article is a valid alternative to the conventional resin embedding-based protocol: it is much cheaper, can be adopted by any histological laboratory, and allows immunohistochemical analysis to be conducted.

  17. [Standardization of Blastocystis hominis diagnosis using different staining techniques].

    Science.gov (United States)

    Eymael, Dayane; Schuh, Graziela Maria; Tavares, Rejane Giacomelli

    2010-01-01

    The present study was carried out from March to May 2008, with the aim of evaluating the effectiveness of different techniques for diagnosing Blastocystis hominis in a sample of the population attended at the Biomedicine Laboratory of Feevale University, Novo Hamburgo, Rio Grande do Sul. On hundred feces samples from children and adults were evaluated. After collection, the samples were subjected to the techniques of spontaneous sedimentation (HPJ), sedimentation in formalin-ether (Ritchie) and staining by means of Gram and May-Grünwald-Giemsa (MGG). The presence of Blastocystis hominis was observed in 40 samples, when staining techniques were used (MGG and Gram), while sedimentation techniques were less efficient (32 positive samples using the Ritchie technique and 20 positive samples using the HPJ technique). Our results demonstrate that HPJ was less efficient than the other methods, thus indicating the need to include laboratory techniques that enable parasite identification on a routine basis.

  18. A novel contrast stain for the rapid diagnosis of pityriasis versicolor: A comparison of Chicago Sky Blue 6B stain, potassium hydroxide mount and culture

    Directory of Open Access Journals (Sweden)

    Nikita Lodha

    2015-01-01

    Full Text Available Background: The mycological study of pityriasis versicolor is usually done by potassium hydroxide (KOH mount and culture. However, KOH mount lacks a color contrast and requires a trained eye to interpret, while culture is difficult to perform, time consuming and has low sensitivity. Chicago Sky Blue 6B (CSB is a new contrast stain that highlights the fungal hyphae and spores, blue against a purplish background. Aims and Objectives: This study was done to compare the utility of a novel contrast stain (CSB stain with KOH mount and culture. Materials and Methods: Skin scrapings from the lesions of 100 clinically diagnosed cases of P. versicolor were subjected to (1 KOH mount and CSB stain for direct microscopic examination and (2 culture using Sabouraud′s dextrose agar. The statistical analysis of CSB stain and culture was done using KOH mount as the reference method, as it is the most commonly performed and practical diagnostic test available for P. versicolor. An interrater reliability analysis using the Cohen′s Kappa statistic was performed to determine consistency (agreement among the different modalities. Observations and Results: Direct microscopy with CSB stain, KOH mount and mycological culture showed positive results in 98 (98%, 92 (92% and 56 (56% patients, respectively. Using KOH mount as the reference method, CSB stain had a sensitivity of 100% which was significantly higher than culture (60.9%. Statistically significant fair agreement was found between CSB stain and KOH mount (94% with κ=0.38, P < 0.001. Negligible agreement was found between CSB stain and culture (66%, κ=0.199, P = 0.001 as well as between KOH mount and culture (64%, κ=0.051, P = 0.107. Conclusion: CSB staining of skin scrapings is the most sensitive method for the diagnosis of pityriasis versicolor. Due to the distinct contrast provided by CSB, it is easy to perform, rapid and qualitatively superior to KOH mount.

  19. An immunohistochemical study of feline endometrial adenocarcinoma.

    Science.gov (United States)

    Gil da Costa, R M; Santos, M; Amorim, I; Lopes, C; Pereira, P Dias; Faustino, A M

    2009-05-01

    Feline endometrial adenocarcinomas are uncommon malignant neoplasms that have to date been poorly characterized. The present immunohistochemical study describes the expression of the pancytokeratins AE1 and AE3, cytokeratin-14, vimentin, alpha-actin, cyclo-oxygenase-2, E-cadherin, beta-catenin, the progesterone receptor, the oestrogen receptor and caveolin-1 within normal feline uterine tissue and tissue from six cats with endometrial adenocarcinoma. Synthesis of cyclo-oxygenase-2 and reduced expression of progesterone receptors may be involved in the neoplastic transformation of feline endometrium. The loss of cellular adhesion that occurs within these tumours does not require down-regulation of E-cadherin expression and nuclear translocation of beta-catenin is not a feature of these neoplasms.

  20. Immunohistochemical Assessment of Expression of Centromere Protein—A (CENPA) in Human Invasive Breast Cancer

    Energy Technology Data Exchange (ETDEWEB)

    Rajput, Ashish B. [Department of Pathology and Molecular Medicine, Queen' s University, Kingston, ON K7L 3N6 (Canada); Hu, Nianping [Cancer Research institute, Queen' s University, Kingston, ON K7L 3N6 (Canada); Varma, Sonal; Chen, Chien-Hung [Department of Pathology and Molecular Medicine, Queen' s University, Kingston, ON K7L 3N6 (Canada); Ding, Keyue [NCIC Clinical Trials Group, Queen' s University, Kingston, ON K7L 3N6 (Canada); Park, Paul C. [Department of Pathology and Molecular Medicine, Queen' s University, Kingston, ON K7L 3N6 (Canada); Chapman, Judy-Anne W. [NCIC Clinical Trials Group, Queen' s University, Kingston, ON K7L 3N6 (Canada); SenGupta, Sandip K. [Department of Pathology and Molecular Medicine, Queen' s University, Kingston, ON K7L 3N6 (Canada); Madarnas, Yolanda [Cancer Research institute, Queen' s University, Kingston, ON K7L 3N6 (Canada); Department of Oncology, Cancer Center of Southeastern Ontario, Kingston, ON K7L 2V7 (Canada); Elliott, Bruce E.; Feilotter, Harriet E., E-mail: feilotth@kgh.kari.net [Department of Pathology and Molecular Medicine, Queen' s University, Kingston, ON K7L 3N6 (Canada)

    2011-12-06

    Abnormal cell division leading to the gain or loss of entire chromosomes and consequent genetic instability is a hallmark of cancer. Centromere protein –A (CENPA) is a centromere-specific histone-H3-like variant gene involved in regulating chromosome segregation during cell division. CENPA is one of the genes included in some of the commercially available RNA based prognostic assays for breast cancer (BCa)—the 70 gene signature MammaPrint{sup ®} and the five gene Molecular Grade Index (MGI{sup SM}). Our aim was to assess the immunohistochemical (IHC) expression of CENPA in normal and malignant breast tissue. Clinically annotated triplicate core tissue microarrays of 63 invasive BCa and 20 normal breast samples were stained with a monoclonal antibody against CENPA and scored for percentage of visibly stained nuclei. Survival analyses with Kaplan–Meier (KM) estimate and Cox proportional hazards regression models were applied to assess the associations between CENPA expression and disease free survival (DFS). Average percentage of nuclei visibly stained with CENPA antibody was significantly higher (p = 0.02) in BCa than normal tissue. The 3-year DFS in tumors over-expressing CENPA (>50% stained nuclei) was 79% compared to 85% in low expression tumors (<50% stained nuclei). On multivariate analysis, IHC expression of CENPA showed weak association with DFS (HR > 60.07; p = 0.06) within our small cohort. To the best of our knowledge, this is the first published report evaluating the implications of increased IHC expression of CENPA in paraffin embedded breast tissue samples. Our finding that increased CENPA expression may be associated with shorter DFS in BCa supports its exploration as a potential prognostic biomarker.

  1. Characterization of atherosclerosis by histochemical and immunohistochemical methods in African grey parrots (Psittacus erithacus) and Amazon parrots (Amazona spp.).

    Science.gov (United States)

    Fricke, Cornelia; Schmidt, Volker; Cramer, Kerstin; Krautwald-Junghanns, Maria-Elisabeth; Dorrestein, Gerry M

    2009-09-01

    The aim of the study was to characterize atherosclerotic changes in African grey parrots (Psittacus erithacus) and Amazon parrots (Amazona spp.) by histochemical and immunohistochemical methods. Samples of the aorta ascendens and trunci brachiocephalici from 62 African grey parrots and 35 Amazon parrots were stained by hematoxylin and eosin and Elastica van Gieson for grading of atherosclerosis in these birds. Four different stages were differentiated. The incidence of atherosclerosis in the examined parrots was 91.9% in African grey parrots and 91.4% in Amazon parrots. To evaluate the pathogenesis in birds, immunohistochemical methods were performed to demonstrate lymphocytes, macrophages, smooth muscle cells, and chondroitin sulfate. According to the missing lymphocytes and macrophages and the absence of invasion and proliferation of smooth muscle cells in each atherosclerotic stage, "response-to-injury hypothesis" seems inapplicable in parrots. Additionally, we found alterations of vitally important organs (heart, lungs) significantly correlated with atherosclerosis of the aorta ascendens.

  2. A Definitive Diagnosis of Primary Hodgkin Lymphoma on Endoscopic Biopsy Material Utilizing in-Depth Immunohistochemical Analysis

    Directory of Open Access Journals (Sweden)

    Garnet Horne

    2007-01-01

    Full Text Available The esophagus and stomach can be primary sites for Hodgkin lymphoma (HL. The pathognomonic feature of HL is the Reed-Sternberg cell. Because these cells can be rare in HL tumours, biopsies obtained via endoscopy are usually inadequate for establishing a definitive diagnosis. A case of a gastroesophageal junction primary HL is presented that was diagnosed on endoscopic biopsy material with the assistance of the immunohistochemical stains PAX5 and MUM1 which verified the presence of Reed-Sternberg cells. The patient was effectively treated for HL and avoided traditional laparotomy or thoracotomy procedures to obtain the diagnosis. This advanced immunohistochemical approach should be the standard in the diagnosis of primary gastrointestinal HL. An endoscopy-based approach should obviate more invasive, open biopsy procedures for establishing HL diagnosis.

  3. CARCINOMA PROSTATE HISTOPATHOLOGY IN NEEDLE BIOPSIES INCLUDING REVISED GLEASON’S GRADING AND ROLE OF IMMUNOHISTOCHEMICAL MARKERS

    Directory of Open Access Journals (Sweden)

    Rema Priyadarsini

    2017-05-01

    Full Text Available BACKGROUND Adenocarcinoma of prostate is the most common form of cancer in men accounting for 29% of cancers in developed nations and the incidence of prostatic cancer is 6.4% in males of Trivandrum District. MATERIALS AND METHODS All prostatic biopsies taken per rectally and stained by haematoxylin and eosin. In suspected cases of malignancy immunohistochemical markers, the AMACR P504S and high molecular weight cytokeratin 34E12 were done. RESULTS The total number of cases studied were 142. The final diagnosis with histomorphological features show that maximum cases were prostatic carcinoma constituting 45.5% of the samples received. CONCLUSION All prostatic carcinomas were graded by revised Gleason’s grade (ISUP 2005 and the use of immunohistochemical markers in arriving at a definite diagnosis in carcinoma prostate was confirmed.

  4. Selection and application of exterior stains for wood

    Science.gov (United States)

    R. Sam. Williams; William C. Feist

    1999-01-01

    Exterior stains for wood protect the wood surface from sunlight and moisture. Because stains are formulated to penetrate the wood surface, they are not prone to crack or peel as can film-forming finishes, such as paints. This publication describes the properties of stains and wood, methods for applying stains, and the expected service life of stains.

  5. [Exogenous tooth discoloration in children: black stains].

    Science.gov (United States)

    Bandon, D; Chabane-Lemboub, A; Le Gall, M

    2011-12-01

    Black-stains are a coloring frequently met in pediatric dentistry. They can be medically diagnosed as 1-mm borders or unfinished lines formed by a dark exogenous substance which follows the gingival festoon of bet coronary (in cervical third of the crown) temporary teeth and permanent, or they can appear in like points or dark spots. They are caused by bacteria anaerobic chromogenous. The dominant responsible species are actinomyces. Blacks-stains are ferrous depots, formed following a chemical interaction on the surface of the tooth between sulphide of hydrogen (under the effect of the anaerobic bacteria which are producing hydrogen) and the iron contained in the saliva (by a healthy diet) or that released by red blood corpuscles (in case of bloody gums). Black-stains are a shape of characteristic dental plaque by its flora with trend to calcify. It contains an insoluble iron salt with a content raised in calcium and in inorganic phosphor. The coloring Black-stain is a mild pathology and has no incidence on the vitality of the tooth. Certainly these spots are unsightly. The dental surgeon in current practice can deprive them. The pediatrician plays a leading role in the diagnosis and advice to parents and patients affected by these stains. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

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

    Science.gov (United States)

    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.

  7. Basal cell carcinoma vs basaloid squamous cell carcinoma of the skin: an immunohistochemical reappraisal.

    Science.gov (United States)

    Webb, David V; Mentrikoski, Mark J; Verduin, Lindsey; Brill, Louis B; Wick, Mark R

    2015-04-01

    Typical cutaneous basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) are morphologically dissimilar. It is well known, however, that poorly differentiated SCC may assume a basaloid phenotype, complicating the histologic distinction between these 2 neoplasms. Selected immunohistochemical stains have been used in the past to aid in that differential diagnosis. In the current study, additional markers were evaluated to determine whether they would be helpful in that regard. Twenty-nine cases of metatypical (squamoid) BCC (MBCC) and 25 examples of basaloid SCC (BSCC) were studied using the antibodies Ber-EP4 and MOC-31 as well as a plant lectin preparation from Ulex europaeus I (UEA-1). The resulting immunostains were interpreted independently by 3 pathologists, and the results showed that MBCCs demonstrated strong and diffuse staining for Ber-EP4 (25/29) and MOC-31 (29/29). In contrast, BSCCs tended to be only sporadically reactive for both markers (4/25 and 1/25 cases, respectively). Labeling for UEA-1 was observed in almost all BSCCs (24/25), but only 6 of 29 cases of MBCC showed limited, focal staining with that lectin. These data suggest that MOC-31 is a useful marker in the specified differential diagnosis, especially when used together with UEA-1. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. Kallikrein-related peptidase expression in odontogenic cysts and tumors: An immunohistochemical comparative study.

    Science.gov (United States)

    Darling, Mark Roger; Woodford, Rebecca; Cuddy, Karl Kevin; Jackson-Boeters, Linda; Hayter, Alyssa; Inkaran, Jeyanth; Diamandis, Eleftherios P; Khan, Zia

    2017-11-01

    The aim of the present study was to profile the expression of human kallikrein (KLK)-related peptidases (KLK) in odontogenic lesions. Paraffin-embedded, formalin-fixed, non-odontogenic (control) and odontogenic lesions were stained for KLK using a standard immunohistochemical technique. The intensity and proportion of epithelial cells stained was scored. Reverse transcription-polymerase chain reaction was utilized to evaluate KLK 1-15 mRNA expression in ameloblastomas. KLK 3, 4, 9, 11, and 14 were present in all lesions. KLK 3 staining was increased in ameloblastomas and keratocystic odontogenic tumors. KLK 5 was present only in Keratocystic odontogenic tumor. KLK 6 was significantly higher in ameloblastomas than in other lesions. For KLK 7, keratocystic odontogenic tumors and nasopalatine duct cysts were significantly different. KLK 6, 8, 10, 11, and 13 were significantly higher in ameloblastomas than in other lesions. KLK 9 was increased in keratocystic odontogenic tumors and dentigerous cysts. The expression of KLK 1, 4, 7, 8, 10, and 12 mRNA was found in ameloblastomas. The results suggested that KLK 6, 8, 10, and 13 could be involved in the progression of ameloblastomas. KLK 10 could have a greater role in odontogenic lesions, rather than non-odontogenic lesions. Future studies aim to define the specific roles of KLK cascades in odontogenic lesions. © 2017 John Wiley & Sons Australia, Ltd.

  9. The histopathological and immunohistochemical characteristics of pigmentary and cystic glaucoma in the Golden Retriever.

    Science.gov (United States)

    Esson, Douglas; Armour, Micki; Mundy, Patricia; Schobert, Charles S; Dubielzig, Richard R

    2009-01-01

    A clinical syndrome comprising the formation of thin-walled cysts within the posterior chamber, proteinaceous exudation, and pigment dispersion, which typically culminates in glaucoma is recognized in the Golden Retriever breed. Although not uncommon, this syndrome has been relatively infrequently documented in the literature, particularly from a histological standpoint. Fifteen globes from Golden Retrievers presented to Eye Care for Animals between 2003 and 2009 were evaluated by routine hematoxylin and eosin (H&E) as well as immunohistochemical staining. Alcian blue, periodic acid Schiff (PAS), Masson's trichrome, Cytokeratin, Vimentin, Neuron Specific Enolase (NSE), S-100, and smooth muscle actin staining were performed. The thin-walled cysts stained positive with Vimentin, NSE, and S-100 in 15/15 globes, consistent with a ciliary body epithelial cellular origin. No globes demonstrated goniodysgenesis. All 15 globes exhibited free pigment within the trabecular meshwork. Little to no inflammatory infiltrate was noted in 15/15 eyes. These findings suggest that the term 'uveitis' may be an inappropriate description of this syndrome.

  10. BRAF mutations in metastatic malignant melanoma: comparison of molecular analysis and immunohistochemical expression.

    Science.gov (United States)

    Ehsani, Laleh; Cohen, Cynthia; Fisher, Kevin E; Siddiqui, Momin T

    2014-10-01

    Melanoma is a complex genetic disease, and multiple genetic alterations have been reported to play a role during disease progression. The dysregulation of BRAF signaling has been shown to affect many molecules that promote the continual progression of melanoma. Oncogenic BRAF expression plays a vital role in promoting cell invasion and metastasis in melanoma. It is also associated with poor prognosis in metastatic melanoma. About 40% to 60% of cutaneous melanomas have BRAF mutations, and 90% of the mutations involve a specific substitution at codon 600 (BRAF V600E). In this study, we compared BRAF (V600E) mutation detection by molecular analysis with BRAF expression by immunohistochemical (IHC) analysis using 2 different antibodies. A total of 25 metastatic malignant melanomas were included in this study. 10 of the 25 (40%) cases were positive by molecular analysis using the COBAS 4800 BRAF V600 Real-time PCR assay, and 18 of the 25 (72%) cases were positive by IHC analysis with 2 different antibodies. All positive cases by molecular analysis were positive by both IHC stains (100%). No false negatives were obtained with either IHC stain. Eight of the 25 (32%) were false-positive by both IHC stains. This study demonstrates that IHC analysis is a very sensitive test for evaluation of BRAF mutations in metastatic malignant melanoma and may be useful as an initial screening test.

  11. Immunohistochemical expression of GLUT-1, GLUT-3, and carbonic anhydrase IX in benign odontogenic lesions.

    Science.gov (United States)

    Vasconcelos, Roseane Carvalho; de Oliveira Moura, Jamile Marinho Bezerra; Lacerda Brasileiro Junior, Vilson; da Silveira, Éricka Janine Dantas; de Souza, Lélia Batista

    2016-10-01

    Some benign odontogenic lesions have a distinct biological behavior with high recurrence rates and local aggressive behavior. To determine whether glucose transporters proteins (GLUT-1 and GLUT-3) and carbonic anhydrase IX (CA IX) are associated with the development of as dentigerous cyst (DC), odontogenic keratocyst (OK), and ameloblastoma (AM), we evaluated the immunohistochemical expression of these proteins in these lesions. Immunoexpression of GLUT-1, GLUT-3, and CA IX was evaluated semiquantitative fields in each of the 20 cases of OK, AM, and DC. The cases were classified according to the scores: 0 (0% positive cells), 1 (50% of positive cells). The statistical analysis was performed using Pearson's chi-square, Kruskal-Wallis and Mann-Whitney tests. All cases were positive for GLUT-1 and 65% of OK showed scored 3. Staining was diffuse in 90% of OK and 85% of DC cases (P GLUT-3. Staining intensity for anhydrase was higher in the epithelium of DC when compared to OK (P = 0.01). Strong staining was observed in 55% of DC and 20% of OK samples (P = 0.01). These results suggest that GLUT-1 may be involved in the metabolic regulation of glucose in odontogenic lesions studied. In addition, CA IX appears to influence the development of AM, OK, and DC which can explain the differences their biological behavior. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  12. Campylobacter enteritis: early diagnosis with Gram's stain.

    Science.gov (United States)

    Ho, D D; Ault, M J; Ault, M A; Murata, G H

    1982-10-01

    Campylobacter jejuni has become one of the most important causes of infectious diarrhea in the United States. We examined the utility of Gram's stain of stool for the rapid presumptive diagnosis of Campylobacter enteritis in a large, urban hospital and found that this test has a sensitivity of 43.5% and a specificity of 99.4%. We believe that Gram's stain of stool could be used to direct the early management of up to one half of patients infected with this pathogen.

  13. EVALUASI DALAM PEMBELAJARAN BAHASA ARAB DI STAIN PAMEKASAN

    Directory of Open Access Journals (Sweden)

    Maimun ---

    2011-07-01

    Full Text Available Evaluating study of arab Ianguage in STAIN Pamekasan represents one of the among study activity chain starts from study planning process, study execution and last is study evaluation. Evaluate at ability target (maharah language which must be evaluated, and also at elements (‘anashir language. As have been tolerated, that in arab Ianguage study at least there are four abilities (maharah which must be mastered by educative participant to get predicate that he is one who have ability in the arab Ianguage field. The Maharah is Maharah Istima'(ability to correct reading, maharah al-Kalam (ability to converse, maharah Kitabah (ability to write, and maharah al-Qiraah (ability to read. Methodologically, study evaluation process of arab Ianguage in STAIN Pamekasan researched by using the qualitative approach with research type of case which its target is all currator lecturers of Arab Ianguage subject. Research result is obtained as follows: a Step of assessment starting from preparation, execution, data-processing and follow-up, b evaluation form intended here is some instruments used by all lecturer to get information about college student efficacy, in STAIN Pamekasan the pattern mentioned becomes two, that is tes form and non tes form, c all lecturers more tend to using one approach of Ianguage tes, that is integrative approach

  14. Blood culture gram stain, acridine orange stain and direct sensitivity-based antimicrobial therapy of bloodstream infection in patients with trauma

    Directory of Open Access Journals (Sweden)

    Behera B

    2010-01-01

    Full Text Available Purpose: The purpose of this study was to ascertain if the simple practice of Gram stain, acridine orange stain and direct sensitivity determination of positive blood culture bottles could be used to guide early and appropriate treatment in trauma patients with clinical suspicion of sepsis. The study also aimed to evaluate the error in interpreting antimicrobial sensitivity by direct method when compared to standard method and find out if specific antibiotic-organism combination had more discrepancies. Findings from consecutive episodes of blood stream infection at an Apex Trauma centre over a 12-month period are summarized. Materials and Methods: A total of 509 consecutive positive blood cultures were subjected to Gram staining. AO staining was done in BacT/ALERT-positive Gram-stain negative blood cultures. Direct sensitivity was performed from 369 blood culture broths, showing single type of growth in Gram and acridine orange staining. Results of direct sensitivity were compared to conventional sensitivity for errors. Results: No ′very major′ discrepancy was found in this study. About 5.2 and 1.8% minor error rates were noted in gram-positive and gram-negative bacteria, respectively, while comparing the two methods. Most of the discrepancies in gram-negative bacteria were noted in β lactam - β lactamase inhibitor combinations. Direct sensitivity testing was not reliable for reporting of methicillin and vancomycin resistance in Staphylococci. Conclusions: Gram stain result together with direct sensitivity testing is required for optimizing initial antimicrobial therapy in trauma patients with clinical suspicion of sepsis. Gram staining and AO staining proved particularly helpful in the early detection of candidaemia.

  15. Might the Masson trichrome stain be considered a useful method for categorizing experimental tendon lesions?

    Science.gov (United States)

    Martinello, Tiziana; Pascoli, Francesco; Caporale, Giovanni; Perazzi, Anna; Iacopetti, Ilaria; Patruno, Marco

    2015-08-01

    Strain injuries of tendons are the most common orthopedic injuries in athletic subjects, be they equine or human. When the tendon is suddenly damaged, an acute inflammatory phase occurs whereas its repetitive overloading may cause chronic injuries. Currently the criteria used for grading injuries are general and subjective, and therefore a reliable grading method would be an improvement. The main purpose of this study was to assess qualitatively the histological pattern of Masson trichrome stain in healthy and injured tendons; indeed, the known "paradox" of Masson staining was used to create an evaluation for the matrix of tendons, following experimental lesions and natural repair processes. A statistically significant difference of aniline-staining between healthy and lesioned tendons was observed. Overall, we think that the Masson staining might be regarded as an informative tool in discerning the collagen spatial arrangement and therefore the histological characteristics of tendons.

  16. Quantification of immunohistochemical findings of neurofibrillary tangles and senile plaques for a diagnosis of dementia in forensic autopsy cases.

    Science.gov (United States)

    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. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  17. Solid acid catalysts: Stain and shine

    Science.gov (United States)

    Chen, Peng

    2011-11-01

    Catalyst particles for fluid catalytic cracking are vital for the oil-refinery industry, but their activity is hard to diagnose because of their inter- and intra-particle structural inhomogeneity. With fluorescence confocal microscopy and selective staining, one can now pinpoint the catalytic activity within single catalyst particles from an industrial reactor.

  18. Autofluorescence of routinely hematoxylin and eosin- stained ...

    African Journals Online (AJOL)

    SERVER

    2008-03-04

    Mar 4, 2008 ... ... Hosokawa S, Nagaike K, Tagawa T (2004). A new immunofluoro-staining method using red fluorescence of PerCP on formalin-fixed paraffin-embedded tissues. J. Immunol. Methods, 293: 143-151. Rotomskis R, Streckyte G (2004). Fluorescence diagnostics of tumors. Medicina (Kaunas), 40: 1219-1230.

  19. A comparative assessment of commonly employed staining ...

    African Journals Online (AJOL)

    1991-03-16

    Mar 16, 1991 ... T. F. H. G. JACKSON, PH.D. V. GATHIRAM, F,C.P.(S.A.). Department of Medical Microbiology, University of Natal,. Durban. J. VAN DEN ENDE, F.F. PATH. (S.A.). A=pted 30 Mat 1990. have similar sizes,-shapes and staining characteristics to yeasts and other coccidia, diagnostic difficulties can be expected.

  20. The Language of Stained-Glass Windows

    Science.gov (United States)

    Brew, Charl Anne

    2010-01-01

    The splendor and beauty of stained glass punctuates any room. In this article, the author describes a cross-curriculum project which incorporated the French classes' research and written study of France in the Middle Ages. For the project the author suggested Sainte-Chapelle which is considered a reliquary and was built by Louis IX to house the…

  1. Corneal staining after treatment with topical tetracycline

    NARCIS (Netherlands)

    Lapid-Gortzak, Ruth; Nieuwendaal, Carla P.; Slomovic, Allan R.; Spanjaard, Lodewijk

    2006-01-01

    PURPOSE: The purpose of this paper is to report a case of corneal staining after treatment with topical tetracycline. METHODS: A patient with crystalline keratopathy caused by Streptococcus viridans after corneal transplantation was treated topically with tetracycline eye drops, based on results of

  2. Photoacoustic Imaging of Port-Wine Stains

    NARCIS (Netherlands)

    Kolkman, R.G.M.; Mulder, M.J.; Mulder, Miranda J.; Glade, Conrad P.; Steenbergen, Wiendelt; van Leeuwen, Ton

    2008-01-01

    Background and Objective: To optimize laser therapy of port-wine stains (PWSs), information about the vasculature as well as lesion depth is valuable. In this study we investigated the use of photoacoustic imaging (PAI) to obtain this information. - Study Design/Materials and Methods: PAI uses

  3. Canker Stain Affects Delaware Sycamores Pest Alert

    Science.gov (United States)

    Alan Iskra; Gary Schwetz; Michael Valenti

    2001-01-01

    An often fatal disease of American sycamore (Platanus occidentalis), known as canker stain, is caused by the fungus, Ceratocystis fimbriata f.sp. platani. This fungus, indigenous to the United States, occurs in urban and forested areas from New Jersey to Georgia and west to Missouri and Louisiana. Other trees affected are the Oriental plane (Platanus orientalis) and...

  4. Comparison of effectiveness of abrasive and enzymatic action of whitening toothpastes in removal of extrinsic stains - a clinical trial.

    Science.gov (United States)

    Patil, P A; Ankola, A V; Hebbal, M I; Patil, A C

    2015-02-01

    To compare the effectiveness of abrasive component (perlite/calcium carbonate) and enzymatic component (papain and bromelain) of whitening toothpaste in removal of extrinsic stains. This study is a randomized, triple blind and parallel group study in which 90 subjects aged 18-40 years were included. At baseline, stains scores were assessed by Macpherson's modification of Lobene Stain Index and subjects were randomly assigned to two groups with 45 subjects in each. Group 1 used whitening toothpaste with enzymatic action and group 2 with abrasive action. After 1 month, stain scores were assessed for the effectiveness of the two toothpastes and 2 months later to check the stain prevention efficacy. Wilcoxson's test was used to compare between baseline 1 and 2 months stain scores, and Mann-Witney U-test was applied for intragroup comparison. The mean baseline total stain score for the subjects allocated to the enzymatic toothpaste was 37.24 ± 2.11 which reduced to 30.77 ± 2.48 in 1 month, and for the abrasive paste, total stain reduced from 35.08 ± 2.96 to 32.89 ± 1.95. The reductions in total stain scores with both the pastes were significant compared with baseline stain scores (at 1 month Group 1, P = 0.0233 and Group 2, P = 0.0324; at 2 months, Group 1 P = 0.0356). Both the toothpastes proved to be equally good in removal of extrinsic stains; however, the enzymatic paste showed better results as compared to abrasive toothpaste. Whitening toothpaste with abrasive action and enzymatic action are equally effective in removal of extrinsic stains; however, whitening toothpaste with abrasive action needs to be used with caution. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  5. Cutaneous Squamous Cell Carcinoma Developing from Recessive Dystrophic Epidermolysis Bullosa: A Case Report and an Immunohistochemical Study

    Directory of Open Access Journals (Sweden)

    Akira Tsukada

    2012-09-01

    Full Text Available We describe a 49-year-old Japanese woman with cutaneous squamous cell carcinoma (SCC developing from recessive dystrophic epidermolysis bullosa (RDEB. Interestingly, immunohistochemical staining revealed dense infiltration of CD163+ M2 macrophages and numerous Foxp3+ regulatory T cells (Tregs around the tumor. Since the contribution of immunosuppressive factors (e.g. TGFβ to the carcinogenesis of SCC from RDEB was recently reported, our present findings suggest one of the possible contributions of immunosuppressive cells, such as CD163+ M2 macrophages and Tregs, to the carcinogenesis of SCC from RDEB.

  6. Vocal fold nodules: morphological and immunohistochemical investigations.

    Science.gov (United States)

    Martins, Regina Helena Garcia; Defaveri, Julio; Custódio Domingues, Maria Aparecida; de Albuquerque E Silva, Rafael; Fabro, Alexandre

    2010-09-01

    The objective of this study was to investigate the morphological and immunohistochemical characteristics of vocal fold nodules. The study design was prospective and retrospective. For the histological study, we reviewed 15 slides from the surgical cases of vocal fold nodules, in which we analyzed epithelium, basal membrane (bm), and lamina propria. For the transmission and scanning electron microscopy (TEM, SEM) studies, five new cases on vocal fold nodules were included. Immunohistochemistry study was carried out in the 15 specimens, using antifibronectin, antilaminin, and anticollagen IV antibodies. The main histological alterations were epithelial hyperplasia (73.33%), basement membrane thickening (86.66%), edema, and fibrosis (93.33%). SEM--reduction in mucous lacing and increase in the desquamating cells, without epithelial erosion. TEM--hyperplasia of the epithelium, enlargement of the intercellular junctions, which was filled by fluid, subepithelial thickening of the lamina reticularis, and break points in the basal membrane. Immunohistochemistry--we identified greater immunoexpression of fibronectin on the basal membrane, on the lamina propria, and around the vessels. Antilaminin and anticollagen IV antibodies showed higher pigmentation on the endothelium of the vessels than that on the basal membrane. In vocal fold nodules, combined assessment using light microscopy, electron microscopy, and immunohistochemistry can reveal important morphological details useful in characterizing these lesions. 2010 The Voice Foundation. Published by Mosby, Inc. All rights reserved.

  7. Immunohistochemical markers of cancerogenesis in the lung.

    Directory of Open Access Journals (Sweden)

    Lech Chyczewski

    2007-06-01

    Full Text Available Lung cancer is the leading cause of cancer deaths for people of both sexes worldwide. Early diagnosis of precancer lesions may be of crucial significance to lowering lung cancer mortality. The World Health Organization has defined three preneoplastic lesions of the bronchial epithelium: squamous dysplasia and carcinoma in situ, atypical adenomatous hyperplasia and diffuse idiopathic pulmonary neuroendocrine cell hyperplasia. These lesions are believed to progress to squamous cell carcinoma, adenocarcinoma and carcinoid tumors, respectively. Apart from WHO classification, two other lesions such as bronchiolization and bronchiolar columnar cell dysplasia (BCCD can be observed and thought to be preneoplastic lesions leading to adenocarcinoma. In this review we summarize the data of morphological and cell cycle related proteins changes in both central and peripheral compartments of lung. Many molecular changes, which accompany the multistep process of the development of invasive types of cancer, may be observed thanks to the application of immunohistochemical markers. A deeper knowledge of molecular and genetic changes accompanying pre-cancer states may show new directions of early diagnostics of cancer development.

  8. Comparing Quantitative Immunohistochemical Markers of Angiogenesis to Contrast-Enhanced Subharmonic Imaging.

    Science.gov (United States)

    Gupta, Aditi; Forsberg, Mark A; Dulin, Kelly; Jaffe, Samantha; Dave, Jaydev K; Halldorsdottir, Valgerdur G; Marshall, Andrew; Forsberg, Anya I; Eisenbrey, John R; Machado, Priscilla; Fox, Traci B; Liu, Ji-Bin; Forsberg, Flemming

    2016-09-01

    Different methods for obtaining tumor neovascularity parameters based on immunohistochemical markers were compared to contrast-enhanced subharmonic imaging (SHI). Eighty-five athymic nude female rats were implanted with 5 × 10(6) breast cancer cells (MDA-MB-231) in the mammary fat pad. The contrast agent Definity (Lantheus Medical Imaging, North Billerica, MA) was injected, and SHI was performed using a modified Sonix RP scanner (Analogic Ultrasound, Richmond, British Columbia, Canada) with a L9-4 linear array (transmitting/receiving frequencies, 8/4 MHz). Afterward, specimens were stained for endothelial cells (CD31), vascular endothelial growth factor (VEGF), and cyclooxygenase 2 (COX-2). Tumor neovascularity was assessed in 4 different ways using a histomorphometry system (×100 magnification: (1) over the entire tumor; (2) in small sub-regions of interest (ROIs); (3) in the tumor periphery and centrally; and (4) in 3 regions of maximum marker expression (so-called hot spots). Results from specimens and from SHI were compared by linear regression. Fifty-four rats (64%) showed tumor growth, and 38 were successfully imaged. Subharmonic imaging depicted the tortuous morphologic characteristics of tumor neovessels and delineated small areas of necrosis. The immunohistochemical markers did not correlate with SHI measures over the entire tumor area or over small sub-ROIs (P > .18). However, when the specimens were subdivided into central and peripheral regions, COX-2 and VEGF correlated with SHI in the periphery (r = -0.42; P = .005; and r = -0.32; P = .049, respectively). When comparing quantitative contrast measures of tumor neovascularity to immunohistochemical markers of angiogenesis in xenograft models, ROIs corresponding to the biologically active region should be used to account for tumor heterogeneity.

  9. Immunohistochemical studies with a monoclonal antibody on the distribution of phosphophoryn in predentin and dentin.

    Science.gov (United States)

    Nakamura, O; Gohda, E; Ozawa, M; Senba, I; Miyazaki, H; Murakami, T; Daikuhara, Y

    1985-09-01

    A monoclonal antibody was raised against phosphophoryn, a unique noncollagenous phosphoprotein in dentin. Mouse myeloma NS-I cells were fused with spleen cells obtained from BALB/c mice immunized with phosphophoryn from fetal calf tooth germs. Mice inoculated with the hybridoma produced ascites fluid containing the antibody and this reacted only with a band of phosphophoryn transblotted from polyacrylamide gel. Immunohistochemical studies with the antibody showed that phosphophoryn was present in odontoblasts, odontoblastic processes and dentin, but not in the matrix of predentin, and that the phosphophoryn content of the dentin layer was high at and around the predentin-dentin junction and gradually decreased toward the enamel layer. The area corresponding to mantle dentin was not stained with the antibody.

  10. Immunohistochemical expression of interleukin 8 in skin biopsies from patients with inflammatory acne vulgaris

    Directory of Open Access Journals (Sweden)

    El Maged Rabee A

    2007-01-01

    Full Text Available Abstract Background This study was conducted to evaluate the immunohistochemical (IHC expression of interleukin 8 (IL-8 in skin biopsies of inflammatory acne vulgaris (IAV in an attempt to understand the disease pathogenesis. Materials and methods A total of 58 biopsies, 29 from lesional IAV and 29 normal non lesional sites were immunostained for IL-8. The intensity of staining was evaluated in the epidermis and dermis and was scored as mild, moderate and severe. The expression was correlated with the clinical grade, disease course and histological changes. Results IL-8 immunoreactivity was expressed in lesional IAV compared to non lesional skin biopsies (p Conclusion We were able to demonstrate altered immunoreactivity of IL-8 in IAV compared to normal skin. Targeted therapy to block IL-8 production may hold promise in limiting the deleterious effects of IL-8-mediated inflammatory response and angiogenesis.

  11. Immunohistochemical analysis of mast cell infiltrates and microvessel density in oral squamous cell carcinoma.

    Science.gov (United States)

    Pyziak, L; Stasikowska-Kanicka, O; Danilewicz, M; Wągrowska-Danilewicz, M

    2013-12-01

    The aim of the study was to evaluate mast cell concentration and microvessel density in perilesional and intralesional regions of oral squamous cell carcinoma (OSCC) and furthermore to assess the possible relationship between the above-mentioned parameters. Paraffin-embedded specimens from 47 cases of OSCC and 12 cases of normal mucosa were investigated immunohistochemically with anti-CD-31 antibody to stain microvessels and anti-tryptase antibody to visualize mast cells. The degree of vascularization and mast cell infiltration was measured with an image analysis system. The study revealed considerably increased microvessel density and mast cell abundance in intralesional and perilesional regions of OSCCs in comparison to normal mucosa. There was a significant positive correlation between microvessel density and mast cell concentration in both localizations of OSCCs (p therapeutic significance which require further research.

  12. Immunohistochemical localization of growth factor-regulated channel (GRC) in human tissues.

    Science.gov (United States)

    Kowase, Takanori; Nakazato, Yoichi; Yoko-O, Hideaki; Morikawa, Akihiro; Kojima, Itaru

    2002-06-01

    Insulin-like growth factor-I activates a calcium-permeable cation channel GRC (growth factor-regulated channel). In the present study, we investigated the immunohistochemical localization of GRC in human tissues using a polyclonal anti-GRC antibody. Immunoreactive GRC was detected in the stomach, duodenum, large intestine and prostate. In these tissues, GRC-expressing cells were distributed solitarily in the epithelium and coexpressed chromogranin-A, a marker of neuroendocrine cells. GRC was also expressed in the epithelium of the pancreatic duct, mammary gland, parotid gland, and submandibular gland. Epithelial cells of the renal tubule and the tracheal gland were also stained with anti-GRC antibody. In the lung, alveolar macrophages expressed GRC. In the brain, Purkinje cells of the cerebellum and arachnoid cells of the meningitis expressed GRC. These results indicate that GRC is expressed in restricted types of cells in particular tissues, and that GRC may modulate calcium entry in these cells.

  13. EGFR, p53, IDH-1 and MDM2 immunohistochemical analysis in glioblastoma: therapeutic and prognostic correlation

    Directory of Open Access Journals (Sweden)

    Richard Murdoch Montgomery

    2015-07-01

    Full Text Available We studied 36 glioblastoma cases at HC-UNICAMP from 2008 to 2012 and classified the immunohistochemical distribution of the wild-type epidermal growth factor receptor (EGFR, mutated forms of p53 protein and isocitrate dehydrogenase-1 (IDH-1 and murine double protein 2 (MDM2. Immunostaining findings were correlated with clinical data and response to treatment (surgery, chemotherapy and radiotherapy. About 97% of the tumors were primary, most of them localized in the frontal lobe. Mean time free of clinical or symptomatic disease and free time of radiological disease were 7.56 and 7.14 months, respectively. We observed a significant positive correlation between expressions of p53 and MDM2, EGFR and MDM2. Clinical, radiological and overall survivals also showed a significant positive correlation. p53 staining and clinical survival showed a significant negative correlation. The current series provides clinical and histopathological data that contribute to knowledge on glioblastoma in Brazilians.

  14. Bone marrow stroma in idiopathic myelofibrosis and other haematological diseases. An immunohistochemical study

    DEFF Research Database (Denmark)

    Lisse, I; Hasselbalch, H; Junker, P

    1991-01-01

    staining reactions with antibodies against type III procollagen (pN collagen), type IV collagen, fragment P1 of laminin and factor VIII. Patients with osteomyelosclerosis had particularly increased collagen content, including both newly deposited type III collagen (pN collagen) and mature collagen fibres....... As in normal bone marrow, argyrophilic fibres and type III collagen displayed a close co-distribution, which was also demonstrated for type IV collagen and laminin. While normal bone marrow sinusoids had discontinuous basement membranes, fibrosing bone marrow was characterized by endothelial cell proliferation......Bone marrow stroma was investigated immunohistochemically in 31 patients with haematological diseases, mainly idiopathic myelofibrosis (n = 8) and related chronic myeloproliferative disorders (n = 14). The bone marrow from patients with idiopathic myelofibrosis and some CML patients showed marked...

  15. Surgical management and immunohistochemical study of corneal plaques in vernal keratoconjunctivitis.

    Science.gov (United States)

    Lin, Hsiu-Yi; Yeh, Po-Ting; Shiao, Cheng-Shiang; Hu, Fung-Rong

    2013-09-01

    Two children with shield ulcer in vernal keratoconjunctivitis unresponsive to steroid therapy received plaque removal by superficial keratectomy, followed by amniotic membrane transplantation (AMT). Hematoxylin and eosin staining of the excised corneal specimen revealed a thick layer of eosinophilic material attached to the Bowman's layer. These deposits were positive for eosinophil granule major basic protein, as confirmed by an immunohistochemical study. The shield ulcer healed after the amniotic membrane was removed. No recurrent corneal plaque developed, although corneal opacity complicated in both cases. Lamellar keratectomy with AMT offers an effective management by removing the cytotoxic plaques and protecting the denuded stroma from deposition of inflammatory debris. Copyright © 2012. Published by Elsevier B.V.

  16. Immunohistochemical detection of S100A1 in the postmortem diagnosis of acute myocardial infarction

    Science.gov (United States)

    2013-01-01

    Background Sudden cardiac death resulting from acute myocardial infarction (AMI) constitutes a significant percentage of the caseload for forensic and clinical pathologists. When sudden death occurs at an early stage (autopsy material, all human cases of definite myocardial infarction and suspected early infarction showed well-defined areas without S100A1 staining. None of the normal human cases showed diffuse depletion of S100A1. Conclusion Our results suggest that immunohistochemical detection of S100A1 is useful for the postmortem diagnosis of AMI at an early stage. Virtual slides The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/4366650979519818 PMID:23683996

  17. The effect of a calcium carbonate/perlite toothpaste on the removal of extrinsic tooth stain in two weeks.

    Science.gov (United States)

    Collins, L Z; Naeeni, M; Schäfer, F; Brignoli, C; Schiavi, A; Roberts, J; Colgan, P

    2005-01-01

    To assess the effect of a calcium carbonate/perlite toothpaste on the levels of extrinsic stain removed at two weeks compared to a silica control toothpaste. In this parallel group, double-blind study, subjects were stratified by natural baseline stain and tobacco use and allocated at random to one of the two study toothpastes. Subjects brushed with their allocated toothpaste twice daily for two weeks before extrinsic tooth stain was again assessed. The study was performed at 4-Front Research UK Limited, Maldon, UK. 152 adult subjects with at least eight assessable incisors/ canines with natural extrinsic tooth stain completed the study. The extrinsic tooth stain on the facial surfaces of incisors and canines was measured using the Macpherson modification of the Lobene Stain Index. Both of the toothpaste groups had significantly less stain after two weeks of use compared to the baseline value (pperlite toothpaste removed significantly more stain over the two week study than the silica control toothpaste (pperlite toothpaste removes more extrinsic stain than a silica control toothpaste.

  18. HURTLE CELLS IMMUNOHISTOCHEMICAL ACTIVITIES IN HASHIMOTO THYROIDITIS PARENCHYMA.

    Science.gov (United States)

    Tsagareli, Z; Kvachadze, T; Melikadze, E; Metreveli, L; Nikobadze, E; Gogiashvili, L

    2016-11-01

    The present study was designed to evaluate the participation and utility of Hǘrtle cells morphological requirment and transformation under Hashimoto autoimmune thyroiditis versus Riedel´s struma. Several markers have been evaluated to detect induced activities of Hǘrtle cells. Study subject - specimens (tissue fragments) collected from TG surgery (thyroidectomy) for mollecular (receptor) diagnosis of Hǘrtle cells activities using routine histological and immunohistochemical samples. 89 cases were selected in Hashimoto thyroiditis diagnosis with Hǘrtle cells history (adenoma and adenomatous grouth of oncocytes). Markers as: TSH receptors, TTF-1, S-100 protein, also anti-TPO and anti-TG levels in blood plasm were detected. It was shown that solid cell claster-nests like agregation of oncocytes and adenomatous growth foci in parafollicular areas with anti-TPO and anti-TG antibodies levels arising while Riedel´s struma shown only large intra- and extra glandular inflammatory proliferative fibrosing process. Large positive expression of TTF-1 and S-100 protein and the negative reaction of TSH receptor factor suggest that Thyroid parenchyma disorganization and mollecular biological atypia with Hǘrtle cells are proceses due to hypothyreoidismus, as well as neuroectodermal cells prominent activities in 70% of Hashimoto cases.

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

    Energy Technology Data Exchange (ETDEWEB)

    Wu Yihang; Song Mengjie; Zhang Xiaoqing; Zhang Yu; Wang Chunyu; Gu Ning [Jiangsu Key Laboratory for Biomaterials and Devices, State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096 (China); Xin Zhuang; Li Suyi, E-mail: zhangyu@seu.edu.cn, E-mail: guning@seu.edu.cn [Zhongda Hospital, Southeast University, Nanjing 210096 (China)

    2011-06-03

    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 Fe{sub 3}O{sub 4} (OA-USPIO) were synthesized by a novel oxidation coprecipitation method in H{sub 2}O/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.

  20. Biomechanical and immunohistochemical properties of meniscal cartilage after high hydrostatic pressure treatment.

    Science.gov (United States)

    Naal, Florian D; Schauwecker, Johannes; Steinhauser, Erwin; Milz, Stefan; von Knoch, Fabian; Mittelmeier, Wolfram; Diehl, Peter

    2008-10-01

    Meniscal allograft processing procedures, in particular gamma irradiation, deteriorate the biomechanical and biological properties of the transplanted tissue. High hydrostatic pressure (HHP) treatment, widely used in food technology to inactivate microorganisms while preserving natural compounds, might serve as a gentle alternative to gamma irradiation in the processing of meniscal allografts. We therefore investigated the effects of HHP treatment on the biomechanical and immunohistochemical properties of meniscal cartilage. Specimens of bovine menisci were treated with HHP for 10 min (20 degrees C) at 300 MPa and 600 MPa. Untreated control samples were left at room temperature and ambient pressure. We performed repetitive cycling indentation-tests to assess the biomechanical properties-in particular the viscoelastic behavior-of HHP treated and untreated meniscal specimens. Immunohistochemical analysis for collagens type I, II, and III and for the proteoglycans versican, aggrecan and for link-protein was performed by immunolabeling cross-sections of untreated and at 600 MPa HHP treated specimens. Comparing untreated and HHP treated meniscal specimens there were no significant differences for all tested biomechanical parameters. All cross-sections of untreated and HHP treated specimens stained positive for the collagens and proteoglycans. We demonstrated that meniscal cartilage can be treated by HHP at levels as high as 600 MPa without affection of the biomechanical and immunochistochemical properties. Therefore, HHP treatment might serve as a gentle alternative to gamma irradiation in the processing of meniscal allografts. Further research is necessary to verificate the present results in vivo. (c) 2008 Wiley Periodicals, Inc.

  1. Immunohistochemical detection of interstitial collagens in bone and cartilage tissue remnants in an infant Peruvian mummy.

    Science.gov (United States)

    Nerlich, A G; Parsche, F; Kirsch, T; Wiest, I; von der Mark, K

    1993-07-01

    We investigated the immunohistochemical presence of various collagen types in bone and cartilage tissue from an infant Peruvian mummy dating between 500 and 1000 A.D. which had been excavated at the necropolis of Las Trancas in the Nazca region in Peru. Following careful rehydration and decalcification of the tissue, the mummy tissue showed morphologically good preservation of the matrix, which could be shown to be composed of various collagen types in a typical pattern. Bone consisted of a collagen I matrix with a small rim of collagen III and V at the endosteal lining and a pericellular collagen V staining around osteocytic holes. In the hypertrophic cartilage of the epiphyseal growth plate, a typical pattern of collagen types II and X could be found. These observations provide evidence that in well-preserved mummy tissue the antigenic determinants of major matrix components are still adequately preserved for an immunohistochemical analysis. This technique may thus be a very helpful tool for the analysis of pathologic processes of historic bone tissue. It may also allow in certain circumstances a distinction between pseudopathologic tissue destruction and pathologic tissue alteration.

  2. Clinical, histopathological and immunohistochemical assessment of human skin field cancerization before and after photodynamic therapy.

    Science.gov (United States)

    Szeimies, R M; Torezan, L; Niwa, A; Valente, N; Unger, P; Kohl, E; Schreml, S; Babilas, P; Karrer, S; Festa-Neto, C

    2012-07-01

    The field cancerization concept in photodamaged patients suggests that the entire sun-exposed surface of the skin has an increased risk for the development of (pre)-malignant lesions, mainly epithelial tumours. Topical photodynamic therapy (PDT) is a noninvasive therapeutic method for multiple actinic keratosis (AK) with excellent outcome. To evaluate the clinical, histological and immunohistochemical changes in human skin with field cancerization after multiple sessions of PDT with methyl-aminolaevulinate (MAL). Twenty-six patients with photodamaged skin and multiple AK on the face received three consecutive sessions of MAL-PDT with red light (37 J cm(-2)), 1 month apart. Biopsies before and 3 months after the last treatment session were taken from normal-appearing skin on the field-cancerized area. Immunohistochemical stainings were performed for TP-53, procollagen-I, metalloproteinase-1 (MMP-1) and tenascin-C (Tn-C). All 26 patients completed the study. The global score for photodamage improved considerably in all patients (P cancerization after multiple sessions of MAL-PDT is proven. The decrease in severity and extent of keratinocyte atypia associated with a decreased expression of TP-53 suggest a reduced carcinogenic potential of the sun-damaged area. The significant increase of new collagen deposition and the reduction of solar elastosis explain the clinical improvement of photodamaged skin. © 2012 The Authors. BJD © 2012 British Association of Dermatologists.

  3. Use of immunohistochemical markers can refine prognosis in triple negative breast cancer

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    Cheang Maggie CU

    2007-07-01

    Full Text Available Abstract Background Basal-like breast cancer has been extensively characterized on the basis of gene expression profiles, but it is becoming increasingly common for these tumors to be defined on the basis of immunohistochemical (IHC staining patterns, particularly in retrospective studies where material for expression profiling may not be available. The IHC pattern that best defines basal-like tumors is under investigation and various combinations of ER, PR, HER2-, CK5/6+ and EGFR+ have been tested. Methods Using datasets from two different hospitals we describe how using different combinations of immunohistochemical patterns has different effects on estimating prognosis at different time intervals after diagnosis. As our baseline, we used two IHC patterns ER-/PR-/HER2-("triple negative phenotype", TNP and ER-/HER2-/CK5/6+ and/or EGFR+ ("core basal phenotype", CBP. Results There was no overall difference in survival between the two hospital-based series, but there was a difference between the TNP and non-TNP groups which was most marked at 3 years (76.8% vs 93.5%, p Conclusion Our findings suggests that CK5/6 and/or EGFR expressing tumor types have a persistently poorer prognosis over the longer term, an observation that may have important therapeutic implications as drugs that target the EGFR are currently being evaluated in breast cancer.

  4. Immunohistochemical evidence of HLA-G expression in extravillous trophoblast invading decidual tissues

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    V. Prada

    2010-01-01

    Full Text Available HLA-G is a non classical HLA I gene product involved in the regulation of implantation and in the immune tolerance during pregnancy, by modulating maternal immune responses at the fetal-maternal interface. In pregnancies lacking immune tolerance and ending in miscarriage, the expression of HLA-G is very probably defective or altered. In order to contribute to further investigations about HLA-G expression in autoimmune miscarriages, we have tried to describe HLA-G immunohistochemical patterns of expression in a homogeneous cohort of normal first trimester choriodecidual specimens, in the three different extravillous trophoblast (EVT populations ("cell islands", "cell columms", and intravascular EVT cells. We expected to demonstrate HLA-G reactivity to the tested antibody (MEMG/01 in all three populations. All choriodecidual specimens were histologically and clinically reviewed to exclude any pathologic finding. Immunohistochemical identification of trophoblast cells in the selected specimens was performed via wellknown immunostains such as Cytokeratin 8/18 (CAM5.2 and NCL-PLp; anti-Ki67 was also used to point out proliferating EVT cells. Then, MEM-G/01 was tested at various dilutions, with or without pretreatment, to find the optimal protocol. As expected, HLA-G specifically stained all three EVT populations, with decreasing reactivity from EVT cell islands to EVT cell columns or intravascular EVTs. The next step will be the investigation of HLA-G pattern of expression in autoimmune aborters.

  5. Antibody Selection for Cancer Target Validation of FSH-Receptor in Immunohistochemical Settings

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    Nina Moeker

    2017-10-01

    Full Text Available Background: The follicle-stimulating hormone (FSH-receptor (FSHR has been reported to be an attractive target for antibody therapy in human cancer. However, divergent immunohistochemical (IHC findings have been reported for FSHR expression in tumor tissues, which could be due to the specificity of the antibodies used. Methods: Three frequently used antibodies (sc-7798, sc-13935, and FSHR323 were validated for their suitability in an immunohistochemical study for FSHR expression in different tissues. As quality control, two potential therapeutic anti-hFSHR Ylanthia® antibodies (Y010913, Y010916 were used. The specificity criteria for selection of antibodies were binding to native hFSHR of different sources, and no binding to non-related proteins. The ability of antibodies to stain the paraffin-embedded Flp-In Chinese hamster ovary (CHO/FSHR cells was tested after application of different epitope retrieval methods. Results: From the five tested anti-hFSHR antibodies, only Y010913, Y010916, and FSHR323 showed specific binding to native, cell-presented hFSHR. Since Ylanthia® antibodies were selected to specifically recognize native FSHR, as required for a potential therapeutic antibody candidate, FSHR323 was the only antibody to detect the receptor in IHC/histochemical settings on transfected cells, and at markedly lower, physiological concentrations (ex., in Sertoli cells of human testes. The pattern of FSH323 staining noticed for ovarian, prostatic, and renal adenocarcinomas indicated that FSHR was expressed mainly in the peripheral tumor blood vessels. Conclusion: Of all published IHC antibodies tested, only antibody FSHR323 proved suitable for target validation of hFSHR in an IHC setting for cancer. Our studies could not confirm the previously reported FSHR overexpression in ovarian and prostate cancer cells. Instead, specific overexpression in peripheral tumor blood vessels could be confirmed after thorough validation of the antibodies used.

  6. Salivary Duct Carcinoma and Invasive Ductal Carcinoma of the Breast: A Comparative Immunohistochemical Study.

    Science.gov (United States)

    Jalaly, Jalal B; Sanati, Souzan; Chernock, Rebecca D; Dibe, Dikson G; El-Mofty, Samir K

    2018-01-04

    Salivary duct carcinoma (SDC) is a high-grade salivary gland malignancy with great morphological resemblance to invasive ductal carcinoma (IDC) of the breast. Rarely, female patients may have a past history of both SDC and IDC. When these patients present with distant metastasis, accurate identification of the primary tumor is particularly difficult. Additionally, rare metastasis of SDC to the breast and IDC to the salivary (parotid) gland can also present a diagnostic challenge. Our aim was to develop an immunohistochemical panel that reliably distinguishes SDC from IDC. We included all SDCs diagnosed from 1989 to 2016 (23 cases) and 29 treatment naïve and histologically similar IDCs. All cases were stained with androgen receptor (AR), estrogen receptor-alpha (ER-α), progesterone receptor (PR), HER-2, CK5/6, p63, and beta-catenin. The great majority (> 90%) of both SDCs and IDCs reacted positively to AR. The main discrepancy in the immunohistochemical profiles was a distinctly different reactivity to ER-α, PR and HER-2. While 28 IDCs (96.6%) reacted positively to ER-α and/or PR, the majority expressing both (82.8%) with a moderate to strong staining intensity, only 2 SDCs expressed ER-α (8.7%) and 5 others expressed PR (21.7%) with only one case expressing both (P value SDC (34.8%) were positive for HER-2 while none of the IDCs were positive (P value SDC from IDC. Positive reactivity to ER-α, PR or both and negative HER-2 favors a diagnosis of IDC while ER-α, PR negative, HER-2 positive tumors are more likely SDC.

  7. Transforming growth factor beta 1 in oral submucous fibrosis: An immunohistochemical study - Understanding the pathogenesis

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    V. V. Kamath

    2014-01-01

    Full Text Available Background: The development of fibrosis is pathognomic in the potentially malignant oral disorder, oral submucous fibrosis (OSF. Strong evidence exists to implicate the chewing of areca nut in the pathogenesis of the lesion. The constituents of areca nut activate several pro-fibrotic cytokines, chiefly transforming growth factor beta 1 (TGF-β1, which leads to an increased deposition and decreased degradation of extracellular matrix and collagen. TGF-β1 probably represents the major pathway in the deposition of collagen fibers in this condition. The present study aims to identify and correlate the expressions of TGF-β1 immunohistochemically on paraffin sections of various stages of OSF. Materials and Methods: The expression of TGF-β1 antibody was detected immunohistochemically using the anti-TGF-β1 mouse monoclonal antibodies (8A11-NovusBio USA on paraffin sections of 58 cases of OSF, 10 cases of normal buccal mucosa tissue and 5 cases of scar tissue. The site, extent, and intensity of expression and quantification of TGF-β1 were noted and a comparative evaluation between various grades of OSF. Scar tissue and normal oral mucosa was made using image analysis software (Jenoptik optical system-ProReg ® Capture Pro 2.8.8 software [2011]. Results: Cells of spinous layer of the epithelium showed more intense staining in all grades of OSF, Grade II showed the highest percentage of expression, same as that of keloid (17% but less than that of normal mucosa (12%. Positive staining was seen around blood vessels, muscles, fibers in the submucosa and perimuscle fibers. Highest expression was in the muscle in Grade III (80% compared with normal oral mucosa (37%. Conclusion: These results suggest that the pathogenesis of OSF and scar/keloid could be linked through the TGF-β1 pathway. Interventions directed at the TGF-beta pathway may hold the key in the future management of this oral potentially malignant condition.

  8. American Tegumentary Leishmaniasis: Effectiveness of an Immunohistochemical Protocol for the Detection of Leishmania in Skin

    Science.gov (United States)

    Alves, Cibele Fontes; Alves, Cintia Fontes; Figueiredo, Maria Marta; Souza, Carolina Carvalho; Machado-Coelho, George Luiz Lins; Melo, Maria Norma; Tafuri, Washington Luiz; Raso, Pedro; Soares, Rodrigo Pedro; Tafuri, Wagner Luiz

    2013-01-01

    Background American tegumentary leishmaniasis (ATL) is endemic in Latin America, where Brazil has over 27 thousand cases per year. The aim of the present study was to develop an immunohistochemical method (IHC) for ATL diagnosis. For this purpose, we used serum from a dog naturally infected with Leishmania (L) infantum (canine hyperimmune serum) as the primary antibody, followed by a detection system with a secondary biotinylated antibody. Methodology Skin samples were obtained from 73 patients in an endemic area of Caratinga, Minas Gerais (MG) State, Brazil all testing positive for ATL with the Montenegro skin test, microscopy, and PCR. Canine hyperimmune serum of a dog naturally infected with Leishmania (L.) infantum was employed as a primary antibody in an immunohistochemical diagnostic method using streptavidin-biotin peroxidase. To assess the specificity of this reaction, IHC assays employing two monoclonal antibodies were carried out. As the polymer-based technology is less time-consuming and labor intensive than the IHC labeled streptavidin-biotin peroxidase method, we compared the two methods for all samples. Results The IHC method detected ATL in 67 of the 73 cases (91.8%). Immunolabeled parasites were primarily detected inside macrophages either in the superficial or the deep dermis. Detection was facilitated by the high contrast staining of amastigotes (dark brown) against the light blue background. A lower detection rate (71.2%) was observed with the both of the monoclonal Leishmania antibodies compared to the canine hyperimmune serum. This may have been due to a non-specific background staining observed in all histological samples rendering positive detection more difficult. The higher efficacy of the canine hyperimmune serum in the IHC method was confirmed by the method using streptavidin-biotin peroxidase as well as that with the polymer-based technology (biotin-avidin-free system). Conclusions The data are encouraging with regard to validating IHC

  9. The effects of antidepressants and pilocarpine on rat parotid glands: an immunohistochemical study

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    Tatiana Maria Folador Mattioli

    2011-01-01

    Full Text Available 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.

  10. Immunohistochemical Assessment of Expression of Centromere Protein—A (CENPA in Human Invasive Breast Cancer

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    Bruce E. Elliott

    2011-12-01

    Full Text Available Abnormal cell division leading to the gain or loss of entire chromosomes and consequent genetic instability is a hallmark of cancer. Centromere protein –A (CENPA is a centromere-specific histone-H3-like variant gene involved in regulating chromosome segregation during cell division. CENPA is one of the genes included in some of the commercially available RNA based prognostic assays for breast cancer (BCa—the 70 gene signature MammaPrint® and the five gene Molecular Grade Index (MGISM. Our aim was to assess the immunohistochemical (IHC expression of CENPA in normal and malignant breast tissue. Clinically annotated triplicate core tissue microarrays of 63 invasive BCa and 20 normal breast samples were stained with a monoclonal antibody against CENPA and scored for percentage of visibly stained nuclei. Survival analyses with Kaplan–Meier (KM estimate and Cox proportional hazards regression models were applied to assess the associations between CENPA expression and disease free survival (DFS. Average percentage of nuclei visibly stained with CENPA antibody was significantly higher (p = 0.02 in BCa than normal tissue. The 3-year DFS in tumors over-expressing CENPA (>50% stained nuclei was 79% compared to 85% in low expression tumors ( 60.07; p = 0.06 within our small cohort. To the best of our knowledge, this is the first published report evaluating the implications of increased IHC expression of CENPA in paraffin embedded breast tissue samples. Our finding that increased CENPA expression may be associated with shorter DFS in BCa supports its exploration as a potential prognostic biomarker.

  11. Histomorphological and immunohistochemical characterization of 172 cutaneous round cell tumours in dogs

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    Marina Rios Araújo

    2012-08-01

    Full Text Available This paper describes the use of a panel of antibodies (CD117, CD3, CD79a, CD45, cytokeratin, vimentin and E-cadherin on formalin-fixed, paraffin-embedded sections of canine cutaneous round cell tumours. Neoplastic tumours were diagnosed by histology and histochemical stains and included 107 mast cell tumours, 31 cutaneous histiocytomas, two localized histiocytic sarcomas, 21 cutaneous lymphomas, three plasma cell tumours, one transmissible venereal tumour and seven unclassified round cell tumours. The histologic diagnosis was modified in 39.5% of the total 172 neoplasms. The staining for CD45 and Ecadherin were variable, and therefore, the final diagnoses of cutaneous histiocytoma and localized histiocytic sarcoma were made based on histology in association with negative results for CD3, CD79a, CD117 and cytokeratin. The cellular origin of unclassified round cell tumours was defined in all cases. Cutaneous B-cell lymphoma and plasma cell tumours were CD79a-positive and could be distinguished from each other by the morphological characteristics. Mast cell tumours and T cell lymphoma were CD117 and CD3 positive, respectively. The positive staining for vimentin and the negative staining for CD3, CD79a, CD117 and cytokeratin favoured the diagnosis of transmissible venereal tumours. Thus, the final diagnosis of cutaneous round cell tumours should be based on the interpretation of immunohistochemical results together with the cellular morphology observed by histology. Therefore, more studies to optimize the specific markers in formalin-fixed, paraffinembedded tissues (especially for histiocytes are required for definitive diagnosis of round cell tumours in dogs.

  12. Novel methods of cytokine detection: Real-time PCR, ELISPOT, and intracellular cytokine staining

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    Eliza Turlej

    2009-05-01

    Full Text Available Cytokines are small hormone-like proteins that play important roles in immune system control. Cytokines regulate the proliferation and differentiation of cells and hematopoiesis and act as mediators in the inflammatory reaction. Changes in cytokine levels are found in many diseases, such as sepsis, bowel inflammatory disease, autoimmune diseases, as well as graft-versus-host disease. Cytokines levels can be detected using in vivo, in vitro, and ex vivo techniques. The level of cytokine produced can be measured by immunoenzymatic test (ELISA in supernatant after cell culture with the addition of stimulant and in plasma by techniques that measure the level of cytokine secretion in cells (e.g. immunohistochemical staining, ELISPOT, and intracellular cytokine staining, and by molecular biological methods (RPA, real-time PCR, in situ hybridization, and Northern blot. Detection of cytokine mRNA in tissues is useful in the direct determination of heterogenic populations of cytokine-producing cells. Nowadays the most frequently used methods for measuring cytokine level are ELISPOT, intracellular cytokine staining with flow cytometry detection, and real-time PCR. These methods have an important clinical role in vaccine efficacy, in viral, bacterial, and verminous diagnostics, and in determining the efficacy of cancer treatment.

  13. E-cadherin as a prognostic marker in human serous carcinomas of the ovary: an immunohistochemical analysis.

    Science.gov (United States)

    Dian, Darius; Brüning, Ansgar; Mylonas, Ioannis

    2011-08-01

    Ovarian cancer is a gynecologic cancer with a high mortality rate, demonstrating the need for effective and reliable tumor markers during the staging and treatment processes. Expression of E-cadherin was immunohistochemically analyzed in 100 ovarian cancer tissue samples. A significant association of E-cadherin expression with histological grading (p = 0.001) and surgical stage (p = 0.020) could be demonstrated. However, the staining intensity of E-cadherin was not significantly associated with progression-free, cause-specific survival or overall survival in serous ovarian carcinomas. The E-cadherin expression was associated with FIGO surgical staging and histological differentiation in serious ovarian carcinomas, suggesting a substantial role in the carcinogenesis of serous ovarian carcinomas. However, although patients with a strong E-cadherin staining intensity had a better prognosis, no statistical significant differences could be observed. Therefore, E-cadherin might not be a useful prognostic tumor marker in serous ovarian carcinomas.

  14. HISTOLOGIC, IMMUNOHISTOCHEMICAL, AND ELECTRON MICROSCOPIC CHARACTERIZATION OF A MALIGNANT IRIDOPHOROMA IN A DWARF BEARDED DRAGON (POGONA HENRYLAWSONI).

    Science.gov (United States)

    de Brot, Simone; Sydler, Titus; Nufer, Lisbeth; Ruetten, Maja

    2015-09-01

    A dwarf bearded dragon (Pogona henrylawsoni) was presented with a white subcutaneous mandibular mass and multiple nodules in the oral mucosa, heart, liver, kidney, intestine, and visceral fat. Histologically, the tumor consisted of densely packed spindle-shaped cells with brow intracytoplasmic pigment that exhibited white-blue birefringence with polarized light. Immunohistochemical staining was negative for S-100 and weakly positive with melan A. Electron microscopic examination revealed cytoplasmic irregular and oblong empty spaces, laminated and often arranged into short stacks, compatible with reflecting platelet profiles typically seen in iridophores. However, in unstained ultrathin sections, electron-dense crystalline material was present, which filled the empty spaces described for stained sections before. Based on histology, immunohistochemistry, and biologic behavior, a malignant iridophoroma was diagnosed. To the authors' knowledge, iridophoromas in lizards have rarely been characterized by using electronic microscopy. Moreover, this is the first description of an iridophoroma in a dwarf bearded dragon.

  15. Immunohistochemical characterization of lymphocytes in microscopic colitis.

    Science.gov (United States)

    Göranzon, C; Kumawat, A K; Hultgren-Hörnqvist, E; Tysk, C; Eriksson, S; Bohr, J; Nyhlin, N

    2013-11-01

    Microscopic colitis (MC), encompassing the subgroups collagenous colitis (CC) and lymphocytic colitis (LC), is characterized by macroscopically normal or near-normal colonic mucosa, and an increased number of intraepithelial lymphocytes (IELs) and mononuclear cell infiltration in the underlying lamina propria (LP), in addition to an increased collagen layer in CC. This study aimed to characterize the inflammatory cells involved in mucosal inflammation, using immunohistochemistry. Paraffin-embedded biopsies from 23 untreated patients with MC (CC=13, LC=10) and 17 controls were stained with antibodies against CD3, CD4, CD8, CD20, CD30, Foxp3, CD45RO and Ki67. Computerized image analysis was used to calculate areas of stained lymphocytes in the surface and crypt epithelia as well as in the LP. In CC and LC, an increase of predominantly CD8(+) lymphocytes was seen in both the epithelium and the lamina propria, whereas a decreased amount of CD4(+) lymphocytes was found in the lamina propria. CD45RO(+) and Foxp3(+) cells were more abundant in all areas in both patient groups compared to controls, as were CD20(+) areas, although more scarce. Ki67(+) areas were only more abundant in the epithelium, whereas CD30(+) areas were more abundant in the lamina propria of both patient groups compared to controls. This study confirms an increased amount of CD8(+) lymphocytes in the epithelium. Lymphocytic proliferation and activation markers were more abundant, whereas a decreased amount of CD4(+) lymphocytes was seen in the LP. Further studies are needed to reveal the underlying mechanism(s). Copyright © 2013 European Crohn's and Colitis Organisation. Published by Elsevier B.V. All rights reserved.

  16. 21 CFR 864.1850 - Dye and chemical solution stains.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Dye and chemical solution stains. 864.1850 Section... solution stains. (a) Identification. Dye and chemical solution stains for medical purposes are mixtures of synthetic or natural dyes or nondye chemicals in solutions used in staining cells and tissues for diagnostic...

  17. Immunohistochemical localization of epidermal growth factor in rat and man

    DEFF Research Database (Denmark)

    Poulsen, Steen Seier; Nexø, Ebba

    1986-01-01

    Epidermal growth factor (EGF) is a peptide which stimulates cell mitotic activity and differentiation, has a cytoprotective effect on the gastroduodenal mucosa, and inhibits gastric acid secretion. The immunohistochemical localization of EGF in the Brunner's glands and the submandibular glands...

  18. Newer applications of the histological stain prepared from Pterocarpus santalinus.

    Science.gov (United States)

    Sen Gupta, P C; Mukherjee, A K

    1981-03-01

    A histological stain prepared from the heartwood of Pterocarpus santalinus Linn. has been found to be an excellent nuclear stain for various cells of animal and plant origin. As an elastic tissue stain, the results are comparable to standard elastic tissue stains. The striations of voluntary muscle fibers are well shown. The Nissl granules and fibers of cranial nerves in the pons are visualized. When counterstained with light green, it differentially stains muscle and fibrous tissue. The stain can be used as counterstain with certain histochemical procedures with satisfactory results. The preparation and use of this versatile stain are described.

  19. Osteoclastic giant cell tumor of the pancreas: an immunohistochemical study

    DEFF Research Database (Denmark)

    Dizon, M A; Multhaupt, H A; Paskin, D L

    1996-01-01

    A case of an osteoclastic giant cell tumor of the pancreas is presented. Immunohistochemical studies were performed, which showed keratin (CAM, AE1) and epithelial membrane antigen positivity in the tumor cells. The findings support an epithelial origin for this tumor.......A case of an osteoclastic giant cell tumor of the pancreas is presented. Immunohistochemical studies were performed, which showed keratin (CAM, AE1) and epithelial membrane antigen positivity in the tumor cells. The findings support an epithelial origin for this tumor....

  20. An immunohistochemical, histological, and electron-microscopic study of the human periodontal ligament during orthodontic treatment.

    Science.gov (United States)

    Anastasi, Giuseppe; Cordasco, Giancarlo; Matarese, Giovanni; Rizzo, Giuseppina; Nucera, Riccardo; Mazza, Manuela; Militi, Angela; Portelli, Marco; Cutroneo, Giuseppina; Favaloro, Angelo

    2008-05-01

    The periodontal ligament lies between the hard tissues of alveolar bone and cementum of teeth and serves to anchor the tooth to the alveolus and functions as a cushion between these hard tissues to migrate occlusal force during mastication. This tissue is always exposed to mechanical stress during mastication. When occlusal forces exceed the adaptive capacity of the periodontal ligament, the periodontal ligament tissue will be injured and then occlusal trauma will occur. The different modifications of periodontal ligament during load deformation can be monitored by analysis of the expression of different collagen types and fibronectin, with immunohistochemical techniques, and by morphological study of ligament, with light- and transmission electron-microscopic techniques. The use of continued and light orthodontic force generates a pressure of ligament with ejection of parodontal fluid externally and partial closing of vessels. On these basis we performed a study in order to evaluate periodontal ligament collagen types I and IV and the fibronectin modifications induced by application of a precalibrated orthodontic strength. We integrated these results, with light and transmission electron-microscopic observations, in order to evaluate the morphological modifications of periodontal tissue. Our observations showed that the type I collagen immunofluorescence staining is increased in the pressure side; in the tension side, it shows prior to treatment an increase, and after 72 h of treatment, a diminution of the staining pattern. Type IV collagen staining is reduced in both sides, but increased gradually after 7 days from treatment; finally, fibronectin staining pattern is gradually increased in the pressure side and reduced in the tension side. In light and transmission electron-microscopic observations it is possible to show a reduction of vessels at 72 h from treatment, and an increase of vessels after 7 days from treatment. The Malassez's epithelial residues are

  1. The use of Botulinum toxin-a injection for facial wrinkles: a histological and immunohistochemical evaluation.

    Science.gov (United States)

    El-Domyati, Moetaz; Attia, Sameh K; El-Sawy, Ashraf E; Moftah, Noha H; Nasif, Ghada A; Medhat, Walid; Marwan, Belkais

    2015-06-01

    Botulinum toxin (BTX)-A has been used for years in the reduction of facial wrinkles; however, histological and immunohistochemical changes after its use were not previously investigated. To evaluate histological and immunohistochemical changes after BTX-A injection for facial wrinkles, sixteen volunteers, with wrinkles on the upper third of the face, were subjected to single injection of BTX-A. Skin biopsy specimens were obtained from peri-orbital wrinkle site (crow's feet area) before and after 3 months of BTX-A injection. Using histological and immunohistochemical evaluation coupled with computerized morphometric analysis, measurement of epidermal thickness, wrinkle depth and width as well as quantitative evaluation of collagen types I and III and elastin was performed for skin biopsies. After BTX-A injections, there were significant increase in wrinkle width and granular layer thickness (P 0.05). However, collagen fibers showed better organization and orientation after BTX-A injection. The histological changes observed after BTX-A injection for facial wrinkles may help in better understanding of its mechanism of action. © 2015 Wiley Periodicals, Inc.

  2. Effect of toothpaste with natural calcium carbonate/perlite on extrinsic tooth stain.

    Science.gov (United States)

    Matheson, J R; Cox, T F; Baylor, N; Joiner, A; Patil, R; Karad, V; Ketkar, V; Bijlani, N S

    2004-01-01

    The current study was designed to determine the effect of natural calcium carbonate toothpaste containing Perlite and microgranules (Whitening toothpaste) on extrinsic tooth stain compared to a standard commercial toothpaste formulation with precipitated calcium carbonate (PCC) as abrasive and a commercial toothpaste with dicalcium phosphate dihydrate (DCPD) as abrasive. The toothpastes were evaluated in a double blind, three-cell, stratified (tobacco use; baseline tooth stain level), parallel group design study involving 600 subjects with extrinsic tooth stain. Subjects brushed twice daily with their allocated toothpaste for four weeks. Extrinsic tooth stain was measured using the Macpherson modification of the Lobene stain index. ANCOVA showed significant differences between toothpastes (p=0.037). Subsequent multiple comparisons using pairwise t-tests, showed the Whitening toothpaste to be superior to the DCPD toothpaste (p=0.014) and the PCC toothpaste (p=0.067). When a Box-Cox transformation was made to the data (y0.6) to improve normality, these two differences were more accurately estimated at p=0.004 and p=0.03 respectively. The Whitening toothpaste has been shown to be significantly more effective in tooth stain removal than the two standard commercial toothpaste formulations.

  3. CD31 (JC70) expression in plasma cells: an immunohistochemical analysis of reactive and neoplastic plasma cells.

    Science.gov (United States)

    Govender, D; Harilal, P; Dada, M; Chetty, R

    1997-06-01

    To investigate the immunohistochemical expression of CD31 (JC70) in normal and neoplastic plasma cells. Plasma cells in bone marrow biopsies and extramedullary locations were examined. All extramedullary biopsies were formalin fixed and paraffin embedded. The bone marrow biopsies were fixed in formal acetic acid and embedded in paraffin wax. Twenty multiple myelomas (12 bone marrow and eight extramedullary deposits), 10 extramedullary plasmacytomas, and 30 biopsies with reactive plasma cells (10 bone marrow, 20 extramedullary biopsies) were stained with anti-CD31 (JC70) using the streptavidin-biotin detection system with diaminobenzidine as a chromogen. Antigen retrieval in bone marrow biopsies was achieved by pressure cooking. In all other biopsies, antigen retrieval was achieved by microwave pretreatment. All 20 extramedullary cases with reactive plasma cells showed intense membrane staining. Focal staining was detected in reactive plasma cells in bone marrow biopsies. Five of 10 plasmacytomas showed membrane staining. None of the cases of multiple myeloma, either medullary or extramedullary, showed any immunoreactivity for CD31. CD31, a member of the immunoglobulin supergene family of cell adhesion molecules, is strongly expressed in extramedullary reactive plasma cells, focally in bone marrow reactive plasma cells, and occasionally in extramedullary plasmacytomas.

  4. Immunohistochemical expression of E-cadherin and beta-catenin in ameloblastomas and tooth germs.

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    Alves Pereira, Karuza Maria; do Amaral, Bruna Aguiar; dos Santos, Bruna Rafaela Martins; Galvão, Hébel Cavalcanti; Freitas, Roseana de Almeida; de Souza, Lélia Batista

    2010-03-01

    The aim was to analyze the expression of E-cadherin and beta-catenin in ameloblastomas and tooth germs to determine their roles in cell differentiation processes and invasiveness compared with odontogenesis. Twenty-one ameloblastoma cases (16 solid and 5 unicystic tumors) and 5 tooth germs were submitted to the immunohistochemical detection of E-cadherin and beta-catenin. Immunoreactivity was evaluated using descriptive and semiquantitative analysis, investigating the location and intensity of staining. The Fisher exact test was performed, and P values of germs for both E-cadherin (P = .53; P = .44; respectively) and beta-catenin (P = .12; P = .16; respectively). Nuclear staining of beta-catenin was observed in only 4 cases (3 solid and 1 unicystic tumor). The results showed no differences in the expression of E-cadherin or beta-catenin between tooth germs and solid and unicystic ameloblastomas. The expression of these molecules seems mainly to be related to the process of cell differentiation. Copyright 2010 Mosby, Inc. All rights reserved.

  5. Comparative analysis of cell proliferation ratio in plaque and erosive oral lichen planus: An immunohistochemical study.

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    Redder, C Pramod; Pandit, Siddharth; Desai, Dinkar; Kandagal, V Suresh; Ingaleshwar, Pramod S; Shetty, Sharan J; Vibhute, Nupura

    2014-05-01

    Proliferating cell nuclear antigen (PCNA) is a nuclear protein synthesized in the late G1 and S-phase of the cell cycle. Detection of this protein represents a useful marker of the proliferation status of lesions. This study has been carried out to evaluate the cell proliferation rate in oral lichen planus (OLP) and comparison between plaque and erosive lichen planus, which indicates the potential for malignant transformation. This study was comprised of 64 cases of histologically proven lichen planus, out of which 32 cases of plaque and erosive each was taken. Two sections were taken from each, one for H and E staining to verify histological diagnosis according to Eisenberg criteria, other sections were stained according to super sensitive polymer horse radish peroxidise method for identifying immunohistochemical expression of PCNA. Data were statistically analyzed by Tukey high-range statistical domain test. Statistically significant P value was considered lichen planus, erosive type (66.86%) showed higher expression of PCNA followed by plaque (17.07%). Overall, P value was lichen planus followed by plaque type, which ultimately results in increased rate of malignant transformation. PCNA is a good nuclear protein marker to evaluate the proliferation status of OLP. Out of the two types of lichen planus, erosive type possesses more proliferative ratio and chances of malignant change is more in this type. It emphasizes the importance of long-term follow-up with erosive type when compared with plaque type.

  6. Sarcoglycan complex in masseter and sternocleidomastoid muscles of baboons: an immunohistochemical study

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    G. Cutroneo

    2015-06-01

    Full Text Available 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.

  7. Immunohistochemical investigation of endometrial leukocytes in implantation period in rats with streptozotosin-induced diabetes

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    Emel Nacar

    2016-05-01

    Full Text Available Our first aim was to determine the total leukocyte profile in implantation. Second aim was to detect the changes in uterine leukocyte profile in diabetes, a common accompanying disease. For this purpose 4 groups are formed with Wistar albino rats weighing 250-300 g. Two of the groups were non-diabetic and two of them were diabetic. One of the diabetic and one of the non-diabetic groups were left pregnant. Then uterus tissues of pregnant animals were removed in the 5th and 7th days of pregnancy together with tissues of other two non-pregnant groups. Tissues were analyzed immunohistochemically with antibodies CD45, CD3, CD4, CD8, CD56, CD68 and CD79a. It was revealed that pregnancy increased immune staining of CD68, CD3, CD45 and CD56 in endometrium. In addition it was observed that immune staining density of CD68, CD45 and CD56 decreased in diabetes. In the histopathological examination, significant degeneration was detected in the endometrium of diabetic rats. Diabetes could decrease leukocyte proportions in decidua in early pregnancy periods. Therefore immune cell therapies could be administrated in diabetes related problems of pregnancy.

  8. Sarcoglycan complex in masseter and sternocleidomastoid muscles of baboons: an immunohistochemical study.

    Science.gov (United States)

    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

    2015-06-05

    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.

  9. Histological staining methods preparatory to laser capture microdissection significantly affect the integrity of the cellular RNA

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    Li Ming-Chung

    2006-04-01

    Full Text Available Abstract Background Gene expression profiling by microarray analysis of cells enriched by laser capture microdissection (LCM faces several technical challenges. Frozen sections yield higher quality RNA than paraffin-imbedded sections, but even with frozen sections, the staining methods used for histological identification of cells of interest could still damage the mRNA in the cells. To study the contribution of staining methods to degradation of results from gene expression profiling of LCM samples, we subjected pellets of the mouse plasma cell tumor cell line TEPC 1165 to direct RNA extraction and to parallel frozen sectioning for LCM and subsequent RNA extraction. We used microarray hybridization analysis to compare gene expression profiles of RNA from cell pellets with gene expression profiles of RNA from frozen sections that had been stained with hematoxylin and eosin (H&E, Nissl Stain (NS, and for immunofluorescence (IF as well as with the plasma cell-revealing methyl green pyronin (MGP stain. All RNAs were amplified with two rounds of T7-based in vitro transcription and analyzed by two-color expression analysis on 10-K cDNA microarrays. Results The MGP-stained samples showed the least introduction of mRNA loss, followed by H&E and immunofluorescence. Nissl staining was significantly more detrimental to gene expression profiles, presumably owing to an aqueous step in which RNA may have been damaged by endogenous or exogenous RNAases. Conclusion RNA damage can occur during the staining steps preparatory to laser capture microdissection, with the consequence of loss of representation of certain genes in microarray hybridization analysis. Inclusion of RNAase inhibitor in aqueous staining solutions appears to be important in protecting RNA from loss of gene transcripts.

  10. Effects of a Baking Soda Gum on extrinsic dental stain: results of a longitudinal 4-week assessment.

    Science.gov (United States)

    Soparkar, P; Newman, M B

    2001-07-01

    An evaluation of the effects of ARM & HAMMER DENTAL CARE The Baking Soda Gum (AHDC) on extrinsic dental stain was made in 48 subjects presenting with measurable extrinsic stain. The subjects were randomized to use either the baking soda gum or a non-baking soda placebo gum for 20 minutes twice daily after lunch and dinner while brushing once daily. The procedure of limited brushing was chosen to simulate the level of hygiene normally practiced by participants entering a clinical study. After 4 weeks, the reduction in measurable extrinsic stain in the baking soda gum group was statistically significant (P = .0044) relative to baseline. Statistical analysis of the placebo gum group revealed no significant change in extrinsic stain from baseline. The magnitude of the unadjusted longitudinal reduction in extrinsic stain in the baking soda gum group was 29.7% at 4 weeks.

  11. Immunohistochemical Detections of EGFR Mutations in NSCLC

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    Chang LIU

    2014-09-01

    Full Text Available In recent years, it has been well known that non-small cell lung cancer (NSCLC patients with mutations of epidermal growth factor receptor (EGFR response better to EGFR-tyrosine kinase inhibitor treatment. Although DNA-based assays (e.g. DNA sequencing are the most frequently used and a relatively reliable method to detect EGFR mutations, they are complex, time-consuming and relatively expensive for routine use in clinical laboratories, besides they require high quality tumor samples. In contrast, the immunohistochemistry (IHC methods make up fully for the above shortcomings and can serve as screening tests for EGFR mutations. However, there are many factors that can influence the results of IHC methods, such as different staining procedures, different antigen retrieval solutions and different sets of criteria, etc. Thus the IHC methods for detecting EGFR mutations have not been widely used in clinic and only in the research stage. This article reviews the use of IHC methods by different researchers and further discusses how to make the IHC methods work best for the detection of EGFR mutations.

  12. Production of tissue microarrays, immunohistochemistry staining and digitalization within the human protein atlas.

    Science.gov (United States)

    Kampf, Caroline; Olsson, Ingmarie; Ryberg, Urban; Sjöstedt, Evelina; Pontén, Fredrik

    2012-05-31

    The tissue microarray (TMA) technology provides the means for high-throughput analysis of multiple tissues and cells. The technique is used within the Human Protein Atlas project for global analysis of protein expression patterns in normal human tissues, cancer and cell lines. Here we present the assembly of 1 mm cores, retrieved from microscopically selected representative tissues, into a single recipient TMA block. The number and size of cores in a TMA block can be varied from approximately forty 2 mm cores to hundreds of 0.6 mm cores. The advantage of using TMA technology is that large amount of data can rapidly be obtained using a single immunostaining protocol to avoid experimental variability. Importantly, only limited amount of scarce tissue is needed, which allows for the analysis of large patient cohorts (1 2). Approximately 250 consecutive sections (4 μm thick) can be cut from a TMA block and used for immunohistochemical staining to determine specific protein expression patterns for 250 different antibodies. In the Human Protein Atlas project, antibodies are generated towards all human proteins and used to acquire corresponding protein profiles in both normal human tissues from 144 individuals and cancer tissues from 216 different patients, representing the 20 most common forms of human cancer. Immunohistochemically stained TMA sections on glass slides are scanned to create high-resolution images from which pathologists can interpret and annotate the outcome of immunohistochemistry. Images together with corresponding pathology-based annotation data are made publically available for the research community through the Human Protein Atlas portal (www.proteinatlas.org) (Figure 1) (3 4). The Human Protein Atlas provides a map showing the distribution and relative abundance of proteins in the human body. The current version contains over 11 million images with protein expression data for 12.238 unique proteins, corresponding to more than 61% of all proteins

  13. Isolation, Culture, and Staining of Single Myofibers.

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    Gallot, Yann Simon; Hindi, Sajedah M; Mann, Aman K; Kumar, Ashok

    2016-10-05

    Adult skeletal muscle regeneration is orchestrated by a specialized population of adult stem cells called satellite cells, which are localized between the basal lamina and the plasma membrane of myofibers. The process of satellite cell-activation, proliferation, and subsequent differentiation that occurs during muscle regeneration can be recapitulated ex vivo by isolation of single myofibers from skeletal muscles and culturing them under suspension conditions. Here, we describe an improved protocol to evaluate ex vivo satellite cells activation through isolation of single myofibers from extensor digitorum longus (EDL) muscle of mice and culturing and staining of myofiber-associated satellite cells with the markers of self-renewal, proliferation, and differentiation.

  14. Immunohistochemical Characterization of Fumarate Hydratase (FH) and Succinate Dehydrogenase (SDH) in Cutaneous Leiomyomas for Detection of Familial Cancer Syndromes.

    Science.gov (United States)

    Carter, Cody S; Skala, Stephanie L; Chinnaiyan, Arul M; McHugh, Jonathan B; Siddiqui, Javed; Cao, Xuhong; Dhanasekaran, Saravana M; Fullen, Douglas R; Lagstein, Amir; Chan, May P; Mehra, Rohit

    2017-06-01

    Hereditary leiomyomatosis and renal cell carcinoma (HLRCC) is caused by germline mutations in the FH gene, and is associated with increased incidence of leiomyomas and a potentially aggressive variant of renal cell carcinoma (HLRCC-associated RCC). Absent immunohistochemical expression of fumarate hydratase (FH) has previously been used to diagnose HLRCC-associated RCC, but immunohistochemical staining of leiomyomas is not standard practice. We performed immunohistochemistry (IHC) on whole sections from consecutive cutaneous leiomyomas from our archives to evaluate for both FH and succinate dehydrogenase B expression, in addition to clinicopathologic data collection and review of all hematoxylin and eosin-stained slides for blinded morphologic evaluation of features reported to be seen in HLRCC-associated uterine leiomyomas. Ninety-six cutaneous leiomyomas from 87 patients were identified; 12 of these specimens were from 7 patients with documented HLRCC. FH expression by IHC was absent in 9 specimens and retained in 85 specimens; 2 cases were equivocal with minimal FH expression. Seven of the 9 absent expression specimens were from patients with HLRCC, as were both of the equivocal specimens. The overall sensitivity and specificity of absent FH expression in leiomyomas for detection of patients with HLRCC were 70.0% and 97.6%, respectively. Inclusion of cases classified as equivocal increased sensitivity to 75.0%. Succinate dehydrogenase B expression was retained in 95 specimens and equivocal in 1 specimen. None of the evaluated morphologic features showed any association with leiomyomas in HLRCC. Loss of FH immunohistochemical expression in cutaneous leiomyomas is a sensitive and specific marker for detection of HLRCC, thus suggesting a role for prospective FH IHC in patients with these tumors to screen for HLRCC.

  15. Multi-organ distribution of phosphorylated alpha-synuclein histopathology in subjects with Lewy body disorders.

    Science.gov (United States)

    Beach, Thomas G; Adler, Charles H; Sue, Lucia I; Vedders, Linda; Lue, Lihfen; White Iii, Charles L; Akiyama, Haru; Caviness, John N; Shill, Holly A; Sabbagh, Marwan N; Walker, Douglas G

    2010-06-01

    A sensitive immunohistochemical method for phosphorylated alpha-synuclein was used to stain sets of sections of spinal cord and tissue from 41 different sites in the bodies of 92 subjects, including 23 normal elderly, 7 with incidental Lewy body disease (ILBD), 17 with Parkinson's disease (PD), 9 with dementia with Lewy bodies (DLB), 19 with Alzheimer's disease with Lewy bodies (ADLB) and 17 with Alzheimer's disease with no Lewy bodies (ADNLB). The relative densities and frequencies of occurrence of phosphorylated alpha-synuclein histopathology (PASH) were tabulated and correlated with diagnostic category. The greatest densities and frequencies of PASH occurred in the spinal cord, followed by the paraspinal sympathetic ganglia, the vagus nerve, the gastrointestinal tract and endocrine organs. The frequency of PASH within other organs and tissue types was much lower. Spinal cord and peripheral PASH was most common in subjects with PD and DLB, where it appears likely that it is universally widespread. Subjects with ILBD had lesser densities of PASH within all regions, but had frequent involvement of the spinal cord and paraspinal sympathetic ganglia, with less-frequent involvement of end-organs. Subjects with ADLB had infrequent involvement of the spinal cord and paraspinal sympathetic ganglia with rare involvement of end-organs. Within the gastrointestinal tract, there was a rostrocaudal gradient of decreasing PASH frequency and density, with the lower esophagus and submandibular gland having the greatest involvement and the colon and rectum the lowest.

  16. Transition of Immunohistochemical Expression of E-Cadherin and Vimentin from Premalignant to Malignant Lesions of Oral Cavity and Oropharynx

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    Kafil Akhtar

    2016-05-01

    Full Text Available Objectives: We sought to study the expression of epithelial-to-mesenchymal transition markers E-cadherin and vimentin in precancerous lesions of the oral cavity and oropharynx and to use the specific pattern of expression to predict invasiveness. Methods: This cross-sectional study looked at 87 cases of oral and oropharyngeal lesions obtained between December 2012 and November 2014 in the Department of Pathology, Jawaharlal Nehru Medical College, Aligarh Muslim University, India. Fifty-three biopsies from the buccal mucosa, tongue, and pharynx and 34 resected oral specimens were evaluated for premalignant and malignant lesions using hematoxylin and eosin and immunohistochemical stains. Immunohistochemical expression of epithelial marker E-cadherin and mesenchymal marker vimentin was evaluated wherever possible. Slides were examined for staining pattern (cytoplasmic or membrane, proportion, and intensity of staining of tumor cells. Patients follow-up and therapy related changes were also studied. Results: There were 64 premalignant and 23 malignant cases in our study with 65 (74.7% cases seen in males and 22 (25.3% cases seen in females. The majority of malignant cases, (n = 15; 64.2% were seen in the fifth and sixth decades of life while most of the premalignant lesions (n = 36; 56.4% were seen in the fourth and fifth decade. Amongst the 64 premalignant oral lesions, leukoplakia comprised of 14 cases (21.9%, of which three cases had associated mild to moderate dysplasia. The majority of premalignant lesions showed strong E-cadherin expression and decreased expression of vimentin with negative and weak expression in both dysplasias and carcinoma in situ (p = 0.013. E-cadherin expression was significantly reduced in invasive carcinomas compared to dysplasias and carcinoma in situ and the difference in immunoreactivity was statistically significant (p < 0.050. Vimentin expression increased as the tumor progressed from dysplasias to carcinoma in

  17. Immunohistochemical Differentiation of Plasmacytoid Urothelial Carcinoma From Secondary Carcinoma Involvement of the Bladder.

    Science.gov (United States)

    Borhan, Walaa M; Cimino-Mathews, Ashley M; Montgomery, Elizabeth A; Epstein, Jonathan I

    2017-11-01

    Plasmacytoid urothelial carcinoma (UC) is a rare variant of UC that can histologically mimic metastatic cancer involving the urinary bladder. A total of 45 cases of plasmacytoid UC were collected and reviewed histologically. The following immunohistochemical markers were performed: CDX2; polyclonal carcinoembryonic antigen (p-CEA); gross cystic disease fluid protein 15 (GCDFP-15); mammaglobin; estrogen receptor (ER); progesterone receptor (PR); GATA 3 and uroplakin II. In all cases, the plasmacytoid variant of UC lacked expression of ER and mammaglobin. In contrast, GCPDFP-15, PR, CDX2 and p-CEA showed positive staining in 11 (24.4%), 6 (13.3%), 8 (17.7%), and 22 (48.8%) cases, respectively. GCPDFP-15 was expressed in 4/8 female cases with 1 concurrently focally (+2) expressing PR. GATA 3 and uroplakin II was positive in 37/45 cases (82.2%) and 15/45 (33.3%) cases, respectively. A tissue microarray with 40 cases of infiltrating lobular carcinoma of the breast was stained for uroplakin II, and was negative in all cases. Tissue microarrays with 46 cases of gastric signet ring cell adenocarcinomas were all negative for GCDFP-15, ER, PR, GATA3, uroplakin II, and mammaglobin. A panel of stains including mammaglobin, ER, and uroplakin II is recommended to exclude metastatic lobular breast carcinoma to the bladder in cases where a conventional UC component is not present. Immunohistochemistry for CDX2 and p-CEA cannot be utilized to differentiate signet ring cell adenocarcinoma of the gastrointestinal tract from plasmacytoid UC; GATA3 or uroplakin II immunoreactivity can rule out a gastric primary given their negativity in signet ring cell adenocarcinoma of the stomach.

  18. Immunohistochemical expression of epithelial cell markers in corneas with congenital aniridia and ocular cicatrizing pemphigoid.

    Science.gov (United States)

    Auw-Haedrich, Claudia; Agrawal, Manisha; Gabbert, Helmut Erich; Meyer, Peter; Arnold, Nicole; Reinhard, Thomas

    2011-02-01

    We investigated the immunohistochemical characteristics of corneal specimens in congenital aniridia and pemphigoid using various corneal markers to determine the status of the corneal epithelium. Conjunctivalization was clinically suspected in all corneas. Ten aniridia and seven pemphigoid paraffin-embedded corneal specimens were stained with periodic Schiff reagent (PAS) and antibodies against CK3/12, CK12, CK19, breast cancer resistance protein 1 (BCRP) and p63. Aniridia: six cases contained goblet cells, four were negative. Both groups had cases with (three of six; one of four) and without CK19 positivity and cases with (two of six; three of four) and without p63 positivity. All aniridia cases except two in the goblet cell group were CK3/12- and CK12-positive and BCRP-negative. Pemphigoid: only one of the seven cases contained goblet cells. This case stained positively for CK19, 3/12, 12 and p63 and negatively for BCRP. The other six cases were positive for CK3/12, five of which were positive for CK12; only one case was CK19-positive. Three cases were p63-positive and two BCRP-positive. The CK12 staining was heterogenous in most cases and was often found in the superficial layer. Three different stages of epithelial characteristics were found in congenital aniridia and pemphigoid: (i) CK19-negative and inhomogenous CK12-positive cases indicating epithelium mainly from (partly) CK12-deficient limbal stem cells; (ii) CK19- and/or goblet cell-positive and CK12-positive cases with their epithelia originating from CK12-deficient limbal stem cells and from incursing conjunctival cells; and (iii) CK19-positive and CK12-negative cases consisting of conjunctival cells alone. © 2009 The Authors. Journal compilation © 2009 Acta Ophthalmol.

  19. Immunohistochemical analysis of medullary breast carcinoma autoantigens in different histological types of breast carcinomas

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    Kostianets Olga

    2012-11-01

    Full Text Available Abstract Background On the past decade a plethora of investigations were directed on identification of molecules involved in breast tumorogenesis, which could represent a powerful tool for monitoring, diagnostics and treatment of this disease. In current study we analyzed six previously identified medullary breast carcinoma autoantigens including LGALS3BP, RAD50, FAM50A, RBPJ, PABPC4, LRRFIP1 with cancer restricted serological profile in different histological types of breast cancer. Methods Semi-quantitative immunohistochemical analysis of 20 tissue samples including medullary breast carcinoma, invasive ductal carcinoma, invasive lobular carcinoma and non-cancerous tissues obtained from patients with fibrocystic disease (each of five was performed using specifically generated polyclonal antibodies. Differences in expression patterns were evaluated considering percent of positively stained cells, insensitivity of staining and subcellular localization in cells of all tissue samples. Results All 6 antigens predominantly expressed in the most cells of all histological types of breast tumors and non-cancerous tissues with slight differences in intensity of staining and subcellular localization. The most significant differences in expression pattern were revealed for RAD50 and LGALS3BP in different histological types of breast cancer and for PABPC4 and FAM50A antigens in immune cells infiltrating breast tumors. Conclusions This pilot study made possible to select 4 antigens LGALS3BP, RAD50, PABPC4, and FAM50A as promising candidates for more comprehensive research as potential molecular markers for breast cancer diagnostics and therapy. Virtual slides The virtual slides’ for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1860649350796892

  20. Ki-67 Membranous Staining: Biologically Relevant or an Artifact of Multiplexed Immunofluorescent Staining.

    Science.gov (United States)

    Wang, Dan; Pang, Zhengyu; Clarke, Gina M; Nofech-Mozes, Sharon; Liu, Kela; Cheung, Alison M Y; Filkins, Robert J; Yaffe, Martin J

    2016-07-01

    In the process of developing a multiplex of 8 common breast cancer biomarkers (Her2/neu, estrogen receptor, progesterone receptor, Ki-67, aldehyde dehydrogenase-1, NaK-ATPase, cytokeratin 8/18, and myosin smooth muscle) on a single formalin-fixed paraffin-embedded slide using a sequential staining, imaging, and dye bleaching technology developed by General Electric Company, membranous Ki-67 staining was observed and colocalized with Her2/neu staining. Using immunohistochemistry as gold standards, we discovered that membranous Ki-67 was an artifact caused by the binding of cyanine 5-conjugated rabbit polyclonal Ki-67 antibody to a secondary cyanine 3-conjugated donkey anti-rabbit antibody which was previously applied and bound to rabbit Her2/neu antibody in our multiplexing experiment. After blocking with rabbit serum, a successful protocol for 8 biomarker multiplexing without cross-reactivity of antibodies from the same species was developed.

  1. Digital staining of pathological tissue specimens using spectral transmittance

    Science.gov (United States)

    Bautista, Pinky A.; Abe, Tokiya; Yamaguchi, Masahiro; Yagi, Yukako; Ohyama, Nagaaki

    2005-04-01

    Staining of tissue specimens is a classical procedure in pathological diagnosis to enhance the contrast between tissue components such that identification and classification of these components can be easily performed. In this paper, a framework for digital staining of pathological specimens using the information derived from the L-band spectral transmittance of various pathological tissue components is introduced, particularly the transformation of a Hematoxylin and Eosin (HE) stained specimen to its Masson-Trichrome (MT) stained counterpart. The digital staining framework involves the classification of tissue components, which are highlighted when the specimen is actually stained with MT stain, e.g. fibrosis, from the HE-stained image; and the linear mapping between specific sets of HE and MT stained transmittance spectra through pseudo-inverse procedure to produce the LxL transformation matrices that will be used to transform the HE stained transmittance to its equivalent MT stained transmittance configuration. To generate the digitally stained image, the decisions of multiple quadratic classifiers are pooled to form the weighting factors for the transformation matrices. Initial results of our experiments on liver specimens show the viability of multispectral imaging (MSI) for the implementation of digital staining in the pathological context.

  2. Immunohistochemical expression of tumor antigens MAGE-A1, MAGE-A3/4, and NY-ESO-1 in cancerous and benign prostatic tissue.

    Science.gov (United States)

    Hudolin, Tvrtko; Juretic, Antonio; Spagnoli, Giulio Cesare; Pasini, Josip; Bandic, Daniela; Heberer, Michael; Kosicek, Miljenko; Cacic, Mirjana

    2006-01-01

    To investigate immunohistochemical expression of MAGE-A and NY-ESO-1/LAGE-1, cancer testis antigens in prostate tissues showing evidence of malignant transformation or benign hyperplasia. 112 prostate samples from patients undergoing surgery at the Urology Clinic at the Zagreb Clinical Hospital Center from 1995 to 2003 were investigated in this study. Of these, 92 carcinoma samples were obtained by radical prostatectomy, and 20 benign prostatic hyperplasia samples by transvesical prostatectomy. Three monoclonal antibodies were used for immunohistochemical staining: 77B for MAGE-A1, 57B for multi-MAGE-A and D8.38 for NY-ESO-1 expression. Expression of MAGE-A1 was observed in 10.8% of carcinoma samples, whereas multi-MAGE-A and NY-ESO-1/LAGE-1 stained 85.9% and 84.8% of samples. Immunohistochemical staining was only detectable in the cytoplasm. A significant heterogeneity could be observed within a same tissue sample where areas with strong positivities coexisted with cancer testis antigens negative areas. Interestingly, a majority of 57B positive cases were also found to be D8.38 positive (correlation coefficient r=0.727 (P<0.01)). Cancer testis antigens expression was neither significantly correlated with PSA values nor with Gleason score. In benign prostatic hyperplasia tissues MAGE-A1 expression was detected in 5%, while 57B and D8.38 staining was observed in 15% samples, and in all cases percentages of positive cells were always <10%. Our data underline the peculiar relevance of cancer testis antigens expression in prostate cancers, with potential implications regarding both diagnosis and therapy. Copyright (c) 2005 Wiley-Liss, Inc.

  3. Immunohistochemical analysis of the gingiva with periodontitis of type I plasminogen deficiency compared to gingiva with gingivitis and periodontitis and healthy gingiva.

    Science.gov (United States)

    Kurtulus Waschulewski, Idil; Gökbuget, Aslan Y; Christiansen, Nina M; Ziegler, Maike; Schuster, Volker; Wahl, Gerhard; Götz, Werner

    2016-12-01

    Type I plasminogen deficiency (Plgdef) is an uncommon chronic inflammation of mucous membranes. Gingival enlargements usually proceed with progressive periodontal destruction and tooth-loss. Plasmin(ogen)-independent enzymatic mechanisms for fibrin clearance have already been discussed in the literature. Our primary objective was to verify, immunohistochemically, the occurrence of different enzymatic factors involved in tissue breakdown of inflamed compared to healthy gingiva. Secondly, we tried to find out, if these patients have a similar microbiological profile to the patients with known gingivitis and periodontitis. Immunohistochemical analysis of enzymes elastase, plasminogen (plg), cathepsin G, matrix-metalloproteinase (MMP)-3 and MMP-7 and of glycoprotein fibrinogen were performed with gingival tissues from 3 healthy controls, 8 patients with Plgdef and 3 patients with gingivitis and periodontitis. Furthermore, plaque from 5 patients with plasminogen deficiency were also obtained to determine the microbiological profile. Significantly high numbers of elastase positive leukocytes were detected in all samples. Staining for MMP-3 and MMP-7 was seen in samples with gingivitis and periodontitis with a stronger staining in samples with periodontitis by Plgdef. Fibrinogen was detectable in all samples. Staining for plg was stronger in samples with periodontitis than in other samples. Staining for cathepsin G was weak in gingivitis and periodontitis. Subgingival microbial flora showed elevated colony forming units of Prevotella intermedia/nigrescens, Fusobacterium spp., Eikenella corrodens, Porphyromonas gingivalis and viridans streptococci. Strong staining of elastase, MMP-3 and MMP-7 and weak staining of plg in Plgdef samples supports the plasmin(ogen) - independent fibrin clearance. Similar subgingival microbiological flora was observed in periodontitis with Plgdef as in other periodontal diseases. Further investigations should determine the exact pathomechanism

  4. Morphological Features and Immunohistochemical Expression of p57Kip2 in Early Molar Pregnancies and Their Relations to the Progression to Persistent Trophoblastic Disease

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    Marwa Khashaba

    2017-07-01

    Full Text Available Background Although the morphological features characteristic of products of conception specimens including molar pregnancies are well described, substantial histopathological similarities are observed between the different entities, especially in cases of early pregnancies. Furthermore, there are no current solid criteria that could predict cases with progression to persistent gestational trophoblastic disease. In this study, we aimed to determine the most specific histopathological and immunohistochemical features required for accurate diagnosis that can reliably predict the clinical behavior. Methods Sixty-five cases of products of conception were reviewed clinically and pathologically, and any progression to persistent gestational trophoblastic disease (GTD, if present, was noted. Pathological assessment of the archival material included re-cut sections of 5 μm in thickness, routine staining with hematoxylin and eosin and immunohistochemical staining of p57Kip2. Results Certain histopathological criteria were found to be significant in differentiation between complete hydatidiform mole (CHM and partial hydatidiform mole including villous shape and outline, villous trophoblast hyperplasia, and atypia in extravillous trophoblasts. There were no significant differences in any morphological or immunohistochemical features between cases with or without subsequent development of GTD. Conclusions Histopathological diagnosis of molar pregnancy remains problematic especially in early gestation. Their diagnosis should be stated after a constellation of specific histopathological criteria in order not to miss CHM. p57Kip2 immunohistochemistry is of great value in diagnosis of cases that had equivocal morphology by histopathological examination. However, there were no significant features to predict cases that subsequently developed persistent GTD.

  5. [Histochemical staining using silver salts using a microwave oven].

    Science.gov (United States)

    Balaton, A

    1987-01-01

    Some metallic impregnations--Fontana-Masson, Warthin-Starry, Grocott's methenamine silver, Grimelius' and Dieterle's stains have been modified to use a microwave oven. Microwave bombardment markedly reduces the staining times and produces a cleaner background.

  6. Port wine stain on a child's face (image)

    Science.gov (United States)

    Port wine stains are always present at birth. In an infant, they are flat, pink, vascular lesions. Common locations ... may be present anywhere on the body. Port wine stains may appear in association with other syndromes.

  7. Quest for An Ideal, Simple and Cost-Effective Stain for Morphological Assessment of Sperms.

    Science.gov (United States)

    Lingappa, Hemalatha Anthanahalli; Govindashetty, Abhishek Mandya; Krishnamurthy, Anoosha; Puttaveerachary, Ashok Kagathur; Manchaiah, Sanjay; Shimoga, Indira Channagangappa; Mallaradhya, Sushma Hulikere; Gowda, Sarvesh Ballekoppa Mukunda

    2015-10-01

    Recent alarming trends of a substantial rise in the number of cases of infertility with as many as 30-40% being attributed to male-factor associated causes have created a need for further studies and advancements in semen analysis. Despite the focus on semen analysis over the years, assessment of sperm morphology has not been given due importance although it is a simple, standard and baseline diagnostic modality. It can be used to predict the need and outcome of Artificial Reproductive Techniques such as Invitro Fertilization, Gamete Intra Fallopian Tube Transfer and Intra Cytoplasmic Sperm Injection. To find the ideal, simple and cost-effective basic stain for assessment of sperm morphology in a rural tertiary care set- up where advanced equipment for assessment of sperm morphometry are inaccessible. An updated way of determining sperm shape is called the Kruger's strict morphology method. Keeping this as the standard criterion, we studied semen samples of 62 healthy male subjects using four basic staining techniques and the consensus of four independent observers was tabulated. We found that Haematoxylin and Eosin stain was the best stain for assessment of sperm head morphology. Rapid Papanicolau stain was the most ideal, simple and cost-effective stain for overall assessment of sperm morphology. Sperm morphology assessment remains the baseline necessity for the diagnosis and management of male factor associated infertility when advanced techniques are unavailable, inaccessible or unaffordable.

  8. Innervation of vagina: microdissection and immunohistochemical study.

    Science.gov (United States)

    Song, You Bong; Hwang, Kun; Kim, Dae Joong; Han, Seung Ho

    2009-01-01

    The aim of this study is to elucidate the innervation of human vagina by performing a microdissection and PGP immunohistochemistry study. Seven fresh Korean cadavers were used. In five cadavers, the site at which the nerve entered the vaginal wall was observed by microdissection. Two vaginas were stained with trichrome and protein gene product 9.5 (PGP) immunostain. Terminal nerve branches in the vaginal wall were most dense at the second 1/5 partition from the inferior anterior wall (1.49 +/- 0.51/cm(2)). Its density started decreasing (0.57 +/- 0.09/cm(2) -0.89 +/- 0.46/cm(2)) and became scarce at the fourth and fifth 1/5 partitions from the bottom (0.37 +/- 0.14/cm(2)/3-0/cm(2)). The mucosa of the anterior vaginal wall at the distal 1/3 point (designated 1) was 3.50 +/- 2.06 mm in thickness and was 1.18 +/- 0.20 mm at the proximal point (designated 2). It was 1.57 +/- 0.78 mm at the distal 1/3 point (designated 3) of the lateral wall and 1.43 +/- 0.39 mm at the proximal 1/3 point (designated 4) of the posterior wall. Point 1 was thicker than 2, 3, and 4. The vaginal muscle of 1 (5.59 +/- 2.74 mm) was thicker than 2 (3.19 +/- 0.62 mm), 3 (3.31 +/- 1.31 mm), and 4 (2.98 +/- 0.33 mm). Small nerve fibers were most dense at 1 (6.5 +/- 1.1/mm(2)) and scarce at 4 (3.8 +/- 1.5/mm(2)). Large nerve fibers were most dense at 1 (1.9 +/- 0.9/mm(2)) and scarce at 2 (0.7 +/- 0.7/mm(2)) and 4 (0.7 +/- 1.0/mm(2)). The density of small nerve fibers is 2 approximately 2.9/mm(2) in the muscle and 0.7 approximately 1.5/mm(2) in large nerve fibers. The second 1/5 partition of the distal anterior wall had significantly richer innervation than the surrounding areas. Therefore, we think this 1/5 partition of the distal anterior wall could be so called "G-spot."

  9. Clinical study to compare the effectiveness of a test whitening toothpaste with a commercial whitening toothpaste at inhibiting dental stain.

    Science.gov (United States)

    Claydon, N C A; Moran, J; Bosma, M L; Shirodaria, S; Addy, M; Newcombe, R

    2004-12-01

    A single centre, randomised single-blind, three-way crossover study was performed, to compare the effect of an experimental test toothpaste with a commercially available whitening toothpaste and water control at inhibiting extrinsic stain promoted by repeated chlorhexidine/tea rinses. This study used 23 subjects. During the week before the study the subjects received a prophylaxis to remove all staining, plaque and calculus deposits. On the Monday of the following week subjects returned to the clinic to receive their rinses and to check their dentition was stain free. Under direct supervision at both 09:00 and 13:00 hours they rinsed with either a toothpaste slurry or water control that was repeated daily up to and including the following Thursday. Additionally from the Monday to the Thursday each subject rinsed with a 0.2% chlorhexidine mouthrinse, immediately followed by a rinse with a warm black tea solution. This cycle was repeated hourly eight times throughout the day and on the following days until the Friday. Throughout this period volunteers omitted all other forms of oral hygiene except rinsing with the chlorhexidine mouthwash. On the Friday the level of stain on the teeth and dorsum of tongue was assessed using the Lobene stain index for both stain area and intensity. At the end of each trial period each subject received a thorough prophylaxis to remove all plaque, calculus and staining before starting the second and third period of the study. As expected appreciable amounts of extrinsic stain accumulated on the teeth over each study period. The amount of stain following use of the toothpastes and water control was least with the experimental toothpaste, followed by water control and lastly the commercial whitening paste. For all sites combined there was evidence that the experimental paste was significantly superior to both the commercial paste and water control at reducing stain area (p0.05). In this stain-prevention model the use of an experimental

  10. [Immunohistochemical presentation of fibromodulin during wound healing of periodontal cells].

    Science.gov (United States)

    Qian, Hong; Huang, Qun; Xiao, Yin

    2010-04-01

    To investigate immunohistochemical presentation of fibromodulin in vitro during wound healing of three types of periodontal cells and discuss the role of fibromodulin. Cultured periodontal cells were wounded and followed by observation up to 7 days. The immunohistochemical expression of fibromodulin in periodontal cells was observed. Immunohistochemical localization of fibromodulin during cellular wound healing showed that newly migrating periodontal cells toward the wound area were expressed strongly for fibromodulin at day 1 after wounding; thereafter, fibromodulin expression was noted stronger in newly migrating periodontal cells localized in the central part of the wound area than those around the wound area. These data suggest that fibromodulin may be involved in the early events of wound healing for periodontal cells.

  11. An evaluation of a technique to remove stains from teeth using microabrasion.

    Science.gov (United States)

    Price, Richard B T; Loney, Robert W; Doyle, M Gorman; Moulding, M Brent

    2003-08-01

    Microabrasion using a paste made of acid and pumice is a technique that has been used to remove white, yellow and brown stains from enamel. The authors evaluated the technique by studying the effectiveness of a proprietary microabrasion product. One author used microabrasion to remove white, yellow and brown stains from within the outermost layer of the tooth enamel of 32 subjects. Standardized slides of the teeth were taken before and one week after treatment. Four prosthodontists evaluated the paired images, using a standardized questionnaire and visual analog scales ranging from 1 (no improvement in appearance or stain not removed at all) to 7 (exceptional improvement in appearance or stain totally removed). The evaluators were calibrated and blinded. The evaluators always identified a difference between the pretreatment slides and posttreatment slides; they found no difference between the control slides. In all cases but one (97 percent), the treated teeth had improved in appearance with more uniformity in color. Analysis of variance revealed no differences between evaluator ratings (P = .146). The intraclass correlation coefficient for ratings of individual cases by different evaluators was 0.72, representing a "good" level of correlation of the ratings for improvement of appearance and for stain removal. Mean (+/- standard deviation) ratings were 5.38 (+/- 1.26) for improvement of appearance and 5.06 (+/- 1.26) for stain removal. This study showed that enamel microabrasion could remove stains from within the outermost layer of tooth enamel, thereby improving the appearance of the teeth. This study supports recommendations that enamel microabrasion is an effective, atraumatic method of improving the appearance of teeth with stains in the outermost layer of enamel.

  12. A Digital Staining Algorithm for Optical Coherence Tomography Images of the Optic Nerve Head

    Science.gov (United States)

    Mari, Jean-Martial; Aung, Tin; Cheng, Ching-Yu; Strouthidis, Nicholas G.; Girard, Michaël J. A.

    2017-01-01

    Purpose To digitally stain spectral-domain optical coherence tomography (OCT) images of the optic nerve head (ONH), and highlight either connective or neural tissues. Methods OCT volumes of the ONH were acquired from one eye of 10 healthy subjects. We processed all volumes with adaptive compensation to remove shadows and enhance deep tissue visibility. For each ONH, we identified the four most dissimilar pixel-intensity histograms, each of which was assumed to represent a tissue group. These four histograms formed a vector basis on which we ‘projected' each OCT volume in order to generate four digitally stained volumes P1 to P4. Digital staining was also verified using a digital phantom, and compared with k-means clustering for three and four clusters. Results Digital staining was able to isolate three regions of interest from the proposed phantom. For the ONH, the digitally stained images P1 highlighted mostly connective tissues, as demonstrated through an excellent contrast increase across the anterior lamina cribrosa boundary (3.6 ± 0.6 times). P2 highlighted the nerve fiber layer and the prelamina, P3 the remaining layers of the retina, and P4 the image background. Further, digital staining was able to separate ONH tissue layers that were not well separated by k-means clustering. Conclusion We have described an algorithm that can digitally stain connective and neural tissues in OCT images of the ONH. Translational Relevance Because connective and neural tissues are considerably altered in glaucoma, digital staining of the ONH tissues may be of interest in the clinical management of this pathology. PMID:28174676

  13. Comparison of verdeluz orange G and modified Gallego stains.

    Science.gov (United States)

    Kunche, A; Kiresur, M A; Ananthaneni, A; Guduru, V S; Puneeth, H K; Bagalad, B

    2017-11-21

    Tumors of the oral cavity include combinations of hard and soft tissues that may be difficult to identify using routine hematoxylin and eosin (H & E) staining. Although combination stains can demonstrate hard and soft tissues, trichrome stains, such as VanGieson and Masson, cannot differentiate dental hard tissues, such as dentin, cementum and osteoid. Modified Gallegos (MGS) and verdeluz orange G-acid fuchsin (VOF) stains can differentiate components of teeth. We used 10 tissue sections of decalcified bone and 10 pathologic tissue sections that contained different calcified tissues including peripheral ossifying fibroma, odontoma, central ossifying fibroma and cemento-ossifying fibroma. Sections were stained with H & E, VOF or MGS. H and E stained both hard tissues pink. VOF stained bone purple-red, cementum red and collagen blue. MGS stained bone green-blue, cementum red and collagen blue. VOF staining intensity and differentiation was better than MGS staining. VOF staining demonstrated hard tissue components distinctly and exhibited good contrast with the surrounding connective tissue. VOF also is a simple, single step, rapid staining procedure.

  14. Histological study on the staining potentials of Aqueous extract of ...

    African Journals Online (AJOL)

    Histological study on the staining potentials of Aqueous extract of Ceratonia Siliqua bark. EE Okpidu, AU Okon, GP Oyadonghan, LA Ogbodo, ECN Onyenekwe. Abstract. This study was designed to determine the staining potentials of aqueous extract of Ceratonia Siliqua bark adapted for the first time as a counter stain in ...

  15. Comparism of Various Staining Techniques in the Diagnosis of ...

    African Journals Online (AJOL)

    SITWALA COMPUTERS

    5 and the cyclospora species. Their life cycle requires an external intermediate host, usually an animal, ..... Rigor and Franco though observed a superiority of the MZN stain above the trichrome stain ... of disease in diseased population hence its relevance in diagnosis, was noted to be high in trichrome and auramine stains.

  16. 7 CFR 28.442 - Middling Yellow Stained Color.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Middling Yellow Stained Color. 28.442 Section 28.442... Stained Color. Middling Yellow Stained Color is American Upland cotton which in color is deeper than Middling Tinged Color. [57 FR 34498, Aug. 5, 1992] below color grade cotton ...

  17. 7 CFR 28.441 - Strict Middling Yellow Stained Color.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Strict Middling Yellow Stained Color. 28.441 Section... Strict Middling Yellow Stained Color. Strict Middling Yellow Stained Color is color which is deeper than that of Strict Middling Tinged Color. [57 FR 34498, Aug. 5, 1992] ...

  18. Activity and immunohistochemical localization of porphobilinogen deaminase in rat tissues

    DEFF Research Database (Denmark)

    Jørgensen, P E; Erlandsen, E J; Poulsen, Steen Seier

    2000-01-01

    the activity and the immunohistochemical localization of PBGD in the following tissues of wistar female rats: brain, heart, submandibular gland, liver, kidney, pancreas, ovary, stomach, duodenum, jejunum, ileum, colon and musculature. The PBGD activity varied considerably among the tissues. It was highest...... in the liver, 14 pkat/g, and lowest in the jejunum, 0.7 pkat/g. The immunohistochemical localization of PBGD was studied by antibodies raised against a 40 amino acid synthetic peptide that corresponds to a segment in the C-terminal part of PBGD. The study demonstrated that the PBGD immunoreactivity...

  19. Immunohistochemical evaluation of a malignant phecochromocytoma in a wolfdog.

    Science.gov (United States)

    Sako, T; Kitamura, N; Kagawa, Y; Hirayama, K; Morita, M; Kurosawa, T; Yoshino, T; Taniyama, H

    2001-07-01

    A malignant pheochromocytoma with multiple metastases was diagnosed in a 7-year-old male wolfdog that resulted from a cross between an eastern timber wolf (Canis lupus lycaon) and an Alaskan malamute. A yellowish white neoplastic mass approximately 10 cm diameter was found in the right adrenal gland. The neoplasm penetrated through the wall of the caudal vena cava. A diagnosis of pheochromocytoma was established by histopathologic and immunohistochemical procedures. Immunohistochemically, the neoplastic cells expressed chromogranin A, substance P, synaptophysin, Leu-7, protein gene product 9.5, methionine-enkephalin, S100 protein, and galanin. Multiple metastatic tumors were found in the kidneys, spleen, lungs, heart, and liver.

  20. Systematic immunohistochemical screening for Lynch syndrome in colorectal cancer: a single centre experience of 486 patients.

    Science.gov (United States)

    Zumstein, Valentin; Vinzens, Fabrizio; Zettl, Andreas; Heinimann, Karl; Koeberle, Dieter; von Flüe, Markus; Bolli, Martin

    2016-01-01

    Germline mutations in DNA mismatch repair (MMR) genes MLH1, MSH2, MSH6 and PMS2 cause autosomal dominantly inherited Lynch syndrome. Lynch syndrome patients and their families benefit from life-saving intensive cancer surveillance. Approximately one in 30 colorectal cancers arises in the setting of Lynch syndrome. The aim of this study was to assess the detection rate of Lynch syndrome at our institution after introduction of systematic immunohistochemical screening for MMR deficiency in colorectal cancers from 2011 to 2015. Following the recommendations by the Evaluation of Genomic Applications in Practice and Prevention working group all colorectal cancers were immunohistochemically stained for the presence of MMR proteins MLH1, PMS2, MSH2 and MSH6, independent of clinical criteria. In the case of loss of MLH1, the somatic BRAF mutation V600E was assessed with molecular testing and/or immunohistochemistry. Clinical follow-up of potential Lynch syndrome carriers (patients with tumours showing loss of MLH1 expression with absence of BRAFV600E, loss of PMS2, MSH2 or MSH6) was evaluated. Of all patients (n = 486), loss of MMR protein expression was found in 73 (15.0%) tumours. Twenty-eight (6.0%) were classified as potential Lynch syndrome carriers. Of the genetically tested potential Lynch syndrome carriers (10 out of 28 patients), 40% were first diagnosed with Lynch syndrome. Implementation of systematic immunohistochemistry screening for Lynch syndrome showed that 6% of colorectal cancers were potentially Lynch-syndrome related. Tumour board protocols should systematically contain information on MMR status of all colorectal cancers and, in MMR deficient cases, include clear recommendations for genetic counselling for all potential Lynch syndrome patients.

  1. Glomerular sparing pattern in primary kidney neoplasms: clinical, morphological and immunohistochemical study.

    Science.gov (United States)

    Ronny, Faisal M Huq; Sarungbam, Judy; Zhong, Xuelin; Yusuf, Yasmin; Yang, Ximing; Zhong, Minghao

    2014-01-01

    Glomerular sparing (GS) is defined as a unique growth pattern in which tumor cells replace the majority of renal tubes and overrun intact glomeruli. This phenomenon has been well recognized by pathologists as a typical infiltrative pattern and some studies suggested it was an independent risk factor. Here, we study the clinical, pathological, and immunohistochemical features of primary kidney neoplasms with glomerular sparing pattern. We searched the archives of our pathology department for nephrectomy specimens and reviewed all pathology reports from 2009-2013. We selected cases with tumor and collected clinicopathological information, focusing on re-evaluation of cases with glomerular sparing pattern. To facilitate our study we performed immunohistochemical stains of PAX-8, p63, and InI-1 on selected cases. We selected a total of 204 nephrectomy cases in this study, including 163 cases of renal cell carcinoma; 37 cases of urothelial carcinoma; 4 cases from other categories (Wilms tumor, primary diffuse large B-cell lymphoma, angiolipoma, rhabdoid tumor). Finally, we identified 7 cases of primary kidney tumors with glomerular sparing pattern: 2 cases of clear cell renal cell carcinomas (ccRCC), 1 case of collecting duct carcinoma, 2 cases of urothelial carcinoma (UC), 1 case of diffuse large B-cell lymphoma and 1 case of malignant rhabdoid tumor. The primary kidney tumors with glomerular sparing pattern are rare and incidence in our study is <4% (7/204). There is no specificity for any tumor type, but more commonly seen in high grade UC rather than RCC. It can also be seen in rare neoplasms such as collecting duct carcinoma, lymphoma and malignant rhabdoid tumor. These GS cases need to be recognized as they are often associated with high grade, high stage, large tumor size, and worse prognosis.

  2. Histopathological and immunohistochemical diagnosis of infectious bursal disease in poultry birds

    Directory of Open Access Journals (Sweden)

    J. Singh

    2015-11-01

    Full Text Available Aim: The aim of the present study was to diagnose infectious bursal disease (IBD using gross, histopathological, and immunopathological approaches and to compare efficacy of immunohistochemical techniques with conventional diagnostic techniques. Materials and Methods: A total of 33 samples were collected from the six different poultry farms from Ludhiana and the nearby districts. Upon gross analysis of the necropsied birds, the relevant tissue samples such as bursa, kidney, junction of proventriculus and gizzard, heart, and muscles were then processed for histopathological and immunohistochemical studies. Results: Varied macroscopic changes were noted in bursa, characterized as swollen, hemorrhages to atrophy in size. Nonetheless, hemorrhages over thigh muscles were rarely seen. Histologically, the bursa showed prominent fibrotic and atrophic changes. Rarefaction of bursal follicles with intermittent infiltration of lympho-mononuclear cells with chronic cystic changes was additional changes, considered to be paramount for IBD. Expression and localization of IBD specific viral antigens were noticed mainly intracellular to the rarefied areas of bursal follicle section(s, in conjunction to inner lining of the cystic cavities of affected follicles. In addition, the junction of proventriculus and gizzard, the heart muscle, respiratory ciliated epithelium, and proventriculus also revealed positive expression to IBD virus (IBDV antigen. Advanced immunopathological techniques, i.e., immunofluorescence further testified the evidence of antigen as positive green signal within affected follicles. Further consideration to the reliability of various techniques employed, positive correlation (r=0.64623 was emerged out with conventional pathological scoring. Conclusion: It is concluded that the bursa acts as an organ of choice for demonstrating IBDV antigen for specific diagnosis of disease using immunohistochemistry (IHC, and IHC staining is a precise

  3. The significance of immunohistochemical expression of merlin, Ki-67, and p53 in meningiomas.

    Science.gov (United States)

    Pavelin, Sanda; Bečić, Kristijan; Forempoher, Gea; Tomić, Snježana; Capkun, Vesna; Drmić-Hofman, Irena; Mrklić, Ivana; Lušić, Ivo; Pogorelić, Zenon

    2014-01-01

    Meningiomas are one of the most common CNS tumors whose appearance is closely linked to NF2 gene product merlin. Tumor markers Ki-67 and p53 play established role in tumor progression which should be analyzed in close association with merlin expression. The aim of this study was to investigate the immunohistochemical expression of merlin in meningiomas, correlation with Ki-67 and p53, and to determine the association of these results with histologic grade and subtype. The histologic sections of 170 patients with totally resected meningiomas, between January 2000 and December 2010, were classified according to WHO, immunohistochemically stained for Ki-67, p53, and merlin, and analyzed using light microscope. Ki-67 median was 5.6 times higher in group of patients with negative merlin than in those with positive merlin (P=0.05). Statistically significant correlation of merlin with p53 was found (PMerlin expression between 2 combined groups (meningothelial/secretory and fibroblastic/transitional) was statistically significant (P=0.002). By comparing merlin expression and p53 levels, statistically significant difference was found (P=0.017). In the group with positive merlin and negative p53 as well as positive merlin and low p53, meningothelial/secretory subtypes of meningiomas were more common. In combination of negative merlin and negative p53 as well as negative merlin and high p53, there were more meningiomas of fibroblastic/transitional subtype. There was no statistically significant correlation between merlin and tumor grade (P=0.420). There is undeniable influence of merlin on the development and the proliferative ability of meningioma subtypes. Significant role of p53 pathway was confirmed.

  4. Prognostic Value of Her2/neu Expression in Gastrointestinal Stromal Tumors: Immunohistochemical Study

    Directory of Open Access Journals (Sweden)

    Ahmed M Abd El-Aziz

    2017-02-01

    Full Text Available Background: Gastrointestinal stromal tumor (GIST is a relatively rare type of neoplasms. In Egypt, it represents 2.5% of gastrointestinal tumors and 0.3% of all malignancies. Most of the GISTs develop in the stomach. Aim: To reveal the significance of Her2/neu immunohistochemical expression in GIST and its correlation with other histopathologic parameters and tumor relapse after regular follow-up. Patients and methods: This study is a retrospective and prospective cohort. It included 32 patients with GISTs, who were resectable with no distant metastasis. Immunohistochemical staining by Her2/neu was performed after complete surgical resection of the tumors with preservation of the pseudocapsule. Results: In total, 53.1% of cases were men and 46.9% women. Tumors were classified into low-risk (25%, intermediate-risk (21.9%, and high-risk groups (53.1%. Her2/neu expression was negative in 56.3% of GISTs and positive in 43.7%. Its expression was significantly correlated with risk grade ( P = .04, tumor size ( P = .001, mitotic count ( P = .00, and increased risk of relapse ( P = .00. Furthermore, tumor relapse was significantly correlated with the tumor mitotic counts ( P = .00. Using kappa agreement test, it showed that 4 mitotic counts/50 high-power fields (HPF was the cutoff value with which the tumor might be associated more with relapse, with 83% sensitivity, 70% specificity, and P value of .003. Conclusions: Her2/neu might be used as an independent prognostic marker for tumor recurrence after complete resection of GIST, and the cutoff value of mitotic count that might predict tumor relapse is 4/50 HPF. However, more clinical studies with greater number of cases with fluorescent in situ hybridization integration are recommended.

  5. Prognostic Value of Her2/neu Expression in Gastrointestinal Stromal Tumors: Immunohistochemical Study

    Science.gov (United States)

    Abd El-Aziz, Ahmed M; Ibrahim, Eman A; Abd-Elmoghny, Ashraf; El-Bassiouny, Mohammed; Laban, Zina M; Saad El-Din, Somaia A; Shohdy, Youhanna

    2017-01-01

    Background: Gastrointestinal stromal tumor (GIST) is a relatively rare type of neoplasms. In Egypt, it represents 2.5% of gastrointestinal tumors and 0.3% of all malignancies. Most of the GISTs develop in the stomach. Aim: To reveal the significance of Her2/neu immunohistochemical expression in GIST and its correlation with other histopathologic parameters and tumor relapse after regular follow-up. Patients and methods: This study is a retrospective and prospective cohort. It included 32 patients with GISTs, who were resectable with no distant metastasis. Immunohistochemical staining by Her2/neu was performed after complete surgical resection of the tumors with preservation of the pseudocapsule. Results: In total, 53.1% of cases were men and 46.9% women. Tumors were classified into low-risk (25%), intermediate-risk (21.9%), and high-risk groups (53.1%). Her2/neu expression was negative in 56.3% of GISTs and positive in 43.7%. Its expression was significantly correlated with risk grade (P = .04), tumor size (P = .001), mitotic count (P = .00), and increased risk of relapse (P = .00). Furthermore, tumor relapse was significantly correlated with the tumor mitotic counts (P = .00). Using kappa agreement test, it showed that 4 mitotic counts/50 high-power fields (HPF) was the cutoff value with which the tumor might be associated more with relapse, with 83% sensitivity, 70% specificity, and P value of .003. Conclusions: Her2/neu might be used as an independent prognostic marker for tumor recurrence after complete resection of GIST, and the cutoff value of mitotic count that might predict tumor relapse is 4/50 HPF. However, more clinical studies with greater number of cases with fluorescent in situ hybridization integration are recommended. PMID:28469470

  6. Observations on the right ovary of birds of prey: a histological and immunohistochemical study.

    Science.gov (United States)

    Rodler, D; Stein, K; Korbel, R

    2015-06-01

    In most avian species, only the left ovary and oviduct are developed in the adult bird. Right ovaries and oviducts usually do not mature further after hatching and remain only rudimentary. However, occurrence of a functional right ovary is frequently found in several species of birds of prey. In this study, we investigated the occurrence of the right ovaries and their morphology in these bird species. Four examined wild bird species possessed a right ovary: long-eared owl, common buzzard, sparrow hawk and goshawk. We used histological and immunohistochemical techniques to evaluate structural differences of the gonads and tried to correlate the findings with folliculogenesis and endocrine functions. The right ovaries showed different sizes and shapes. Cytoskeletal elements (tubulin and vimentin) and α-smooth muscle actin have been detected in different structures of the right ovaries, but their staining intensity was weaker compared with the left ovary. This shows that also the right ovary is mechanically able to ovulate. We could also demonstrate the expression of oestrogen receptor α and progesterone receptor in the right ovaries, which indicates that also the right ovary can respond to steroid hormone stimuli. We assume that the expression of steroid hormone receptors in the presumptive gonad is still sufficient to mediate the development of a right ovary in the studied species. We conclude that the expression of steroid hormone receptors in the right ovary is involved in its post-natal development. The histological and immunohistochemical data also imply that in the right ovary, folliculogenesis and ovulation can occur. © 2014 Blackwell Verlag GmbH.

  7. Selfish spermatogonial selection: evidence from an immunohistochemical screen in testes of elderly men.

    Directory of Open Access Journals (Sweden)

    Jasmine Lim

    Full Text Available The dominant congenital disorders Apert syndrome, achondroplasia and multiple endocrine neoplasia-caused by specific missense mutations in the FGFR2, FGFR3 and RET proteins respectively-represent classical examples of paternal age-effect mutation, a class that arises at particularly high frequencies in the sperm of older men. Previous analyses of DNA from randomly selected cadaveric testes showed that the levels of the corresponding FGFR2, FGFR3 and RET mutations exhibit very uneven spatial distributions, with localised hotspots surrounded by large mutation-negative areas. These studies imply that normal testes are mosaic for clusters of mutant cells: these clusters are predicted to have altered growth and signalling properties leading to their clonal expansion (selfish spermatogonial selection, but DNA extraction eliminates the possibility to study such processes at a tissue level. Using a panel of antibodies optimised for the detection of spermatocytic seminoma, a rare tumour of spermatogonial origin, we demonstrate that putative clonal events are frequent within normal testes of elderly men (mean age: 73.3 yrs and can be classed into two broad categories. We found numerous small (less than 200 cells cellular aggregations with distinct immunohistochemical characteristics, localised to a portion of the seminiferous tubule, which are of uncertain significance. However more infrequently we identified additional regions where entire seminiferous tubules had a circumferentially altered immunohistochemical appearance that extended through multiple serial sections that were physically contiguous (up to 1 mm in length, and exhibited enhanced staining for antibodies both to FGFR3 and a marker of downstream signal activation, pAKT. These findings support the concept that populations of spermatogonia in individual seminiferous tubules in the testes of older men are clonal mosaics with regard to their signalling properties and activation, thus fulfilling

  8. Selfish spermatogonial selection: evidence from an immunohistochemical screen in testes of elderly men.

    Science.gov (United States)

    Lim, Jasmine; Maher, Geoffrey J; Turner, Gareth D H; Dudka-Ruszkowska, Wioleta; Taylor, Stephen; Rajpert-De Meyts, Ewa; Goriely, Anne; Wilkie, Andrew O M

    2012-01-01

    The dominant congenital disorders Apert syndrome, achondroplasia and multiple endocrine neoplasia-caused by specific missense mutations in the FGFR2, FGFR3 and RET proteins respectively-represent classical examples of paternal age-effect mutation, a class that arises at particularly high frequencies in the sperm of older men. Previous analyses of DNA from randomly selected cadaveric testes showed that the levels of the corresponding FGFR2, FGFR3 and RET mutations exhibit very uneven spatial distributions, with localised hotspots surrounded by large mutation-negative areas. These studies imply that normal testes are mosaic for clusters of mutant cells: these clusters are predicted to have altered growth and signalling properties leading to their clonal expansion (selfish spermatogonial selection), but DNA extraction eliminates the possibility to study such processes at a tissue level. Using a panel of antibodies optimised for the detection of spermatocytic seminoma, a rare tumour of spermatogonial origin, we demonstrate that putative clonal events are frequent within normal testes of elderly men (mean age: 73.3 yrs) and can be classed into two broad categories. We found numerous small (less than 200 cells) cellular aggregations with distinct immunohistochemical characteristics, localised to a portion of the seminiferous tubule, which are of uncertain significance. However more infrequently we identified additional regions where entire seminiferous tubules had a circumferentially altered immunohistochemical appearance that extended through multiple serial sections that were physically contiguous (up to 1 mm in length), and exhibited enhanced staining for antibodies both to FGFR3 and a marker of downstream signal activation, pAKT. These findings support the concept that populations of spermatogonia in individual seminiferous tubules in the testes of older men are clonal mosaics with regard to their signalling properties and activation, thus fulfilling one of the

  9. A systematic review and meta-analysis of immunohistochemical biomarkers that differentiate chromophobe renal cell carcinoma from renal oncocytoma.

    Science.gov (United States)

    Ng, Keng Lim; Morais, Christudas; Bernard, Anne; Saunders, Nicholas; Samaratunga, Hemamali; Gobe, Glenda; Wood, Simon

    2016-08-01

    Numerous immunohistochemical (IHC) biomarkers have been employed to aid in the difficult differentiation between chromophobe renal cell carcinoma (chRCC) and renal oncocytoma (RO). A systematic review and meta-analysis of the published literature was carried out to summarise and analyse the evidence for discriminatory IHC biomarkers to differentiate the two entities. PubMed database was used to identify relevant literature. Primary end point was comparison of positive immunostaining of the biomarkers in chRCC and RO, with extracted data used to calculate OR and 95% CI and statistical I(2) test of heterogeneity for multiple studies. One hundred and nine manuscripts were available for review. Data extracted were subjected to quantitative meta-analysis. Ten most effective biomarkers (OR of chRCC/RO and CI) are: amylase α1A (n=129, OR=0.001, 95% CI 0.0001 to 0.019); Wnt-5a (n=38, OR=0.0076, 95% CI 0.0004 to 0.015); FXYD2 (n=57, OR=130, 95% CI 14.2 to 1192.3); ankyrin-repeated protein with a proline-rich region (ARPP) (n=25, OR=0.0054, 95% CI 0.0002 to 0.12); cluster of differentiation 63 (CD63) (n=62, diffuse (chRCC) vs apical/polar (RO) stain pattern); transforming growth factor β 1 (TGFβ1) (n=34, membranous (chRCC) vs cytoplasmic (RO)); cytokeratin 7 (CK7) (11 studies, n=448, pooled OR=44.22, 95% CI 22.52 to 86.64, I(2)=15%); S100A1 (4 studies, n=124, pooled OR=0.01, 95% CI 0 to 0.03, I(2)=0%); caveolin-1 (2 studies, n=102, pooled OR=32.95, 95% CI 3.67 to 296.1, I(2)=70%) and claudin-7 (3 studies, n=89, pooled OR=24.7, 95% CI 6.28 to 97.1, I(2)=0%). We recommend a panel of IHC biomarkers of amylase α1A, Wnt-5a, FXYD2, ARPP, CD63, TGFβ1, CK7, S100A1, caveolin-1 and claudin-7 to aid in the differentiation of chRCC and RO. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

  10. Immunohistochemical Expression of Estrogen and Progesterone Receptors in Epulis Fissuratum

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    Maryam Seyedmajidi

    2013-01-01

    Full Text Available Background: Epulis Fissuratum (Epulis Fissuratum (EF or Denture Epulis or inflammatory fibrous hyperplasia is a common hyperplastic tumor-like lesion with reactive nature, related to loose and ill-fitting, full or partial removable dentures and it is more common in women than men. For this reason, hormonal influences may also play role in its creation. The effect of steroid hormones especially sex hormones (Estrogen and progesterone on oral mucosa is identified in some studies. In the present study, the distribution pattern and presence of estrogen and progesterone receptors in epithelial, stromal, endothelial and inflammatory cells in Epulis Fissuratum was investigated. Materials and Methods: This cross-sectional study was carried out on 30 samples of paraffin blocks with Epulis Fissuratum diagnosis and 30 samples of normal mucosal tissues as a control group who have had surgery as a margin beside the above lesions and had been obtained from the oral and maxillofacial pathology departement of Babol Dental School since 2003 up to 2010. Intensity of staining and immunoreactivity were evaluated using subjective index and considering the positive control group (breast carcinoma.Results: Epithelial, stromal, endothelial and inflammatory cells didn’t show reaction with monoclonal antibodies against estrogen and progesterone in none of the samples. Conclusion: It seems that the hypothesis of the existence of estrogen and progesterone receptors in epulis fissuratum and normal oral mucosa is ruled out. The possibility of direct effect of estrogen and progesterone in occurring of epulis fissuratum is rejected.

  11. Histopathologic and Immunohistochemical Correlates of Confocal Descriptors in Pigmented Facial Macules on Photodamaged Skin.

    Science.gov (United States)

    Gómez-Martín, Ignacio; Moreno, Sara; Andrades-López, Evelyn; Hernández-Muñoz, Inma; Gallardo, Fernando; Barranco, Carlos; Pujol, Ramon M; Segura, Sonia

    2017-08-01

    Pigmented facial macules on photodamaged skin are a clinical, dermoscopic, and histopathologic challenge. To clinically and dermoscopically characterize, by means of reflectance confocal microscopy (RCM), ambiguous pigmented facial macules and establish a correlation between RCM, histopathologic, and immunohistochemical findings. A prospective study of ambiguous pigmented facial macules on photodamaged skin was conducted in a tertiary referral center for dermatology between January 1, 2009, and December 31, 2015. Sixty-one patients with 63 ambiguous pigmented facial macules and 12 control photodamaged facial areas were included in the study. Melanocyte density in 1-mm basal layers was determined in skin biopsy specimens from all lesions stained with hematoxylin-eosin and immunohistochemical markers (melan-A, microphthalmia-associated transcription factor, and SRY-related HMG-box gene 10). Dermoscopic, RCM images, and histopathologic preparations were systematically evaluated for the presence of lentigo maligna (LM) criteria. Confocal evaluation was blinded to clinical and dermoscopic diagnosis. Sensitivity and specificity of RCM for LM diagnosis and κ value to establish correlations between dermoscopy, RCM, and histopathology were performed. Sensitivity and specificity of RCM for LM diagnosis. Of the 61 patients included in the study, 31 (51%) were women; mean (SD) age was 71.8 (13.1) years. Twenty-four of the 63 (38%) lesions were diagnosed as LM or LM melanoma (LMM) and 39 (62%) as benign pigmented lesions. Reflectance confocal microscopy enhanced the diagnosis of pigmented facial macules with 91.7% sensitivity and 86.8% specificity. Multivariate analysis showed 2 dermoscopic and 2 confocal features associated with LM or LMM: (1) asymmetric follicular pigmentation and targetlike structures, and (2) round, large pagetoid cells and follicular localization of atypical cells, respectively. Continuous proliferation of atypical melanocytes was found in 21 (88%) LM or

  12. Relationship Between Dual Time Point FDG PET and Immunohistochemical Parameters in Preoperative Colorectal Cancer: Preliminary Study

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    Lee, Jai Hyuen; Lee, Won Ae; Park, Seok Gun; Park, Dong Kook; Namgung, Hwan [Dankook Univ. College of Medicine, Cheonan (Korea, Republic of)

    2012-03-15

    The clinical availability of 2 deoxy 2 [18F] fluoro D glucose (FDG) dual time point positron emission tomography/computerized tomography (DTPP) has been investigated in diverse oncologic fields. The aim of this preliminary study was to evaluate the relationship between various immunohistopathologic markers reflecting disease progression of colorectal cancer and parameters extracted from FDG DTPP in colorectal cancer patients. Forty seven patients with histologically confirmed colorectal cancer were analyzed in this preliminary study. FDG DTPP consisted of an early scan 1 h after FDG injection and a delayed scan 1.5 h after the early scan. Based on an analysis of FDG DTPP, we estimated the maximum standardized uptake value (SUV) of tumors on the early and delayed scans (SUV{sup earlya}nd SUV{sup delayed,} respectively). The retention index (RI) was calculated as follows: (SUV{sup delayed-} SUV{sup early)} x 100/ SUV{sup early.} The clinicopathological findings (size and T and N stages) and immunohistochemical factors [glucose transporter 1 (GLUT 1), hexokinase 2 (HK 2), p53, P504S, and {beta} catenin] were analyzed by visual analysis. The RIs calculated from the SUVs ranged from -1.8 to 73.4 (31.8{+-}15.5). The RIs were significantly higher in patients with high T stages (T3 and T4) than with low T stages (T1 and T2; P<0.05). Among the immunohistochemical analytic markers, GLUT 1 had the highest positive staining rate (93.6%) compared to other markers. Based on unvariable analysis, it was shown that the RI of high level GLUT 1 expression was significantly higher than low level GLUT 1 expression (p=0.01), and the RI of high level p53 expression (p=0.08). Multivariate analysis to investigate a link between RI and clinico pathologic parameters of colorectal carcinoma showed that GLUT 1, p53, and T staging were independently connected with increased RIs (p<0.05, total) using backward selection methods. There was no significant statistical relationship between SUV

  13. Central odontogenic fibroma: Report of case with immunohistochemical study

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    Savas Iordanidis

    2013-01-01

    Conclusion: Our immunohistochemical findings, and especially the positive expression of vimentin from the epithelial cells of COF suggests that these cells are primordial. Last but not least, the presence of a relative small number of myofibroblasts in the stroma justifies the non-aggressive behavior of the neoplasm and supports that a part of stromal collagen of COF is produced by these cells.

  14. Immunohistochemical detection of Her-2/neu overexpression in ...

    African Journals Online (AJOL)

    Immunohistochemical detection of Her-2/neu overexpression in breast carcinoma in Nigerians: A 5-year retrospective study. ... cases of invasive lobular carcinoma (10.8%), three cases of medullary carcinoma (3.6%), two cases of papillary carcinoma (2.4%), and a case each of mucinous and clear cell carcinoma (1.2%).

  15. Immunohistochemical expression of tumor antigens MAGE-A1, MAGE-A3/4, and NY-ESO-1 in squamous cell carcinoma of the penis.

    Science.gov (United States)

    Hudolin, Tvrtko; Juretic, Antonio; Pasini, Josip; Tomas, Davor; Spagnoli, Giulio Cesare; Heberer, Michael; Dimanovski, Jordan; Kruslin, Bozo

    2006-07-01

    To investigate by immunohistochemistry the expression of MAGE-A and NY-ESO-1/LAGE-1, cancer testis antigens (CTAs), in squamous cell carcinoma of the penis. A total of 30 penile carcinoma samples from patients undergoing penile amputation at the Urology Clinics at the Zagreb Clinical Hospital Center and University Hospital "Sestre milosrdnice" from 1997 to 2004 were investigated in this study. Three monoclonal antibodies were used for immunohistochemical staining: 77B specific for MAGE-A1, 57B recognizing multiple MAGE-A CTAs, and D8.38, specific for NY-ESO-1 antigen. The expression of MAGE-A1 was not observed in the carcinoma samples, but both multi-MAGE-A and NY-ESO-1-specific reagents stained 29 (97%) of 30 samples. Immunohistochemical staining was prevailingly detected in the cytoplasm. A significant heterogeneity was observed within the same specimen, in which areas with strong positivity coexisted with CTA-negative areas. The extent of CTA expression did not correlate significantly with tumor grade. The results of this study have documented for the first time the expression of CTAs in squamous cell carcinoma of the penis. Additional research is warranted to explore the potential implications regarding both diagnosis and therapy.

  16. Immunohistochemical detection of a unique protein within cells of snakes having inclusion body disease, a world-wide disease seen in members of the families Boidae and Pythonidae.

    Directory of Open Access Journals (Sweden)

    Li-Wen Chang

    Full Text Available Inclusion body disease (IBD is a worldwide disease in captive boa constrictors (boa constrictor and occasionally in other snakes of the families Boidae and Pythonidae. The exact causative agent(s and pathogenesis are not yet fully understood. Currently, diagnosis of IBD is based on the light microscopic identification of eosinophilic intracytoplasmic inclusion bodies in hematoxylin and eosin stained tissues or blood smears. An antigenically unique 68 KDa protein was identified within the IBD inclusion bodies, called IBD protein. A validated immuno-based ante-mortem diagnostic test is needed for screening snakes that are at risk of having IBD. In this study, despite difficulties in solubilizing semi-purified inclusion bodies, utilizing hybridoma technology a mouse anti-IBD protein monoclonal antibody (MAB was produced. The antigenic specificity of the antibody was confirmed and validated by western blots, enzyme-linked immunosorbent assay, immuno-transmission electron microscopy, and immunohistochemical staining. Paraffin embedded tissues of IBD positive and negative boa constrictors (n=94 collected from 1990 to 2011 were tested with immunohistochemical staining. In boa constrictors, the anti-IBDP MAB had a sensitivity of 83% and specificity of 100% in detecting IBD. The antibody also cross-reacted with IBD inclusion bodies in carpet pythons (Morelia spilota and a ball python (python regius. This validated antibody can serve as a tool for the development of ante-mortem immunodiagnostic tests for IBD.

  17. Immuno-histochemical localization of cholesterol binding proteins in ...

    African Journals Online (AJOL)

    SAM

    2014-07-09

    Jul 9, 2014 ... Positive control tissue sections were stained with Sudan Black-B for microscopic visualization of cholesterol .... 6 cm) by holes made. .... Sudan black-B specially stained the cholesterol droplets in form of black granules, which gave positive binding of cholesterol at appropriate binding sites (Figure 8).

  18. Maturity of pericytes in cerebral neocapillaries induced by growth factors: fluorescence immuno-histochemical analysis using confocal laser microscopy.

    Science.gov (United States)

    Jariyapongskul, Amporn; Nakano, Atushi; Yamaguchi, Saburo; Nageswari, Kolammal; Niimi, Hideyuki

    2003-01-01

    The maturity of pericytes in cerebral neocapillaries induced by two different growth factors: basic fibroblast growth factor (bFGF) and platelet-derived growth factor (PDGF), was examined using an immunohistochemical staining technique. Cerebral angiogenesis was induced in mice by implanting a sandwich system of bFGF/PDGF gel and nylon-mesh over the exposed cortex. On 28th day after incubation, a small volume of cerebral tissue with the nylon-mesh was isolated and stained using tetramethyl rhodamine isothiocyanate (TRITC)-labeled secondary antibody to the primary antibody against NG_2 proteoglycan and fluorescein isothiocyanate (FITC)-labeled Griffonia simplicifolia (GS)-lectin. Using a confocal laser microscopic system, we observed the cerebral neocapillaries on the upper surface of the nylon-mesh and evaluated the maturity of pericytes stained with NG_2 based on the fluorescence immunohistological images. The pericyte appeared rich in neocapillaries induced by PDGF. It was suggested that pericytes might play a key role in the regulation of blood flow in neovessels.

  19. Immunohistochemical analysis of cytokeratin expression in reactive eccrine syringofibroadenoma-like lesion: a comparative study with eccrine syringofibroadenoma.

    Science.gov (United States)

    Ohnishi, T; Watanabe, S; Nomura, K

    2000-04-01

    In addition to solitary eccrine syringofibroadenoma (ESFA), there is another type of ESFA which is associated with underlying dermatoses (reactive ESFA-like lesion). Five lesions in 4 patients of reactive ESFA-like lesion were analyzed by an immunohistochemical method using 13 kinds of anti-cytokeratin (CK) antibodies. Two cases of solitary ESFA were also studied by the same procedure for comparison. Suprabasal staining pattern of AE1 and MNF116, which stain CKs 6, 16 and 17, markers of hyperproliferative state, was observed diffusely in 5 lesions of reactive ESFA-like lesions except for focal negative staining in one case, and was observed focally in one case of solitary ESFA. Furthermore, differentiation-specific cytokeratin expression was reduced in 3 of 5 lesions of reactive ESFA-like lesions. Both ESFA and reactive ESFA showed basically similar immunoreactivity suggesting differentiation toward the dermal duct. The above slight difference in immunoreactivity between both lesions may be explained due to inflammatory infiltrates associated with underlying dermatoses.

  20. Neurotoxicity of 1,3,5-trinitrobenzene (TNB): immunohistochemical study of cerebrovascular permeability.

    Science.gov (United States)

    Chandra, A M; Campbell, G A; Reddy, G; Qualls, C W

    1999-05-01

    1,3,5-Trinitrobenzene (TNB) is a soil and water contaminant at certain military installations. Encephalopathy in rats given 10 daily oral doses of TNB has been reported. The lesion was bilaterally symmetric vacuolation and microcavitation in the cerebellar roof nuclei, vestibular nuclei, olivary nuclei, and inferior colliculi. The contribution of the blood-brain barrier (BBB) in the genesis of these lesions remains uncertain. One of the main goals of the present work was to evaluate the functional state of the BBB. Male Fischer 344 rats (five rats/group) were euthanatized after four, five, six, seven, eight, or 10 daily doses of TNB (71 mg/kg). A different set of rats (five rats/group) was allowed to recover for 10 or 30 days after receiving 10 doses of TNB. Integrity of the BBB was assessed by immunohistochemical staining for extravasated plasma albumin on paraffin-embedded sections. Rats euthanatized after four to eight doses had no lesions, and albumin extravasation in the susceptible regions of the brain was minimal. Rats receiving 10 daily doses of TNB had bilaterally symmetric vacuolation and microcavitation in the cerebellar nuclei, vestibular nuclei, and inferior colliculi in association with multifocal, often confluent foci of extravasated albumin in susceptible nuclei. Albumin was present in vascular walls, extracellular space, and neurons. Immunoreactivity in neurons was of two types: cytoplasmic staining representing pinocytic uptake and homogeneous staining of the entire neuron (nucleus and cytoplasm) due to uncontrolled albumin leakage through the damaged cell membrane. In rats allowed to recover for 10 days, the microcavitated foci were infiltrated by glial and gitter cells. Albumin immunoreactivity was present as extracellular granular debris, and neuronal staining (for albumin) was mild. In rats allowed to recover for 30 days, immunoreactivity to albumin was not seen. This study demonstrates that TNB-mediated tissue damage is accompanied by

  1. Electrostatic control of the coffee stain effect

    Science.gov (United States)

    Wray, Alex; Papageorgiou, Demetrios; Sefiane, Khellil; Matar, Omar

    2013-11-01

    The ``coffee stain effect,'' as first explained by Deegan et al. 1997, has received a great deal of attention amongst modellers and experimentalists in recent years, perhaps due in part to its obvious casual familiarity. However, it maintains interest because of its intriguing reliance on an interplay of a trio of effects: contact line pinning, inhomogeneous mass flux, and resulting capillarity-driven flow. What is more, the effect, and especially its suppression or reversal, find applications in fields as diverse as sample recovery, mass spectroscopy and the printing of Organic LEDs. We examine the motion a nanoparticle-laden droplet deposited on a precursor film, incorporating the effects of capillarity, concentration-dependent rheology, together with a heated substrate and resultant mass flux and Marangoni effects. We allow the substrate to act as an electrode and incorporate a second electrode above the droplet. The potential difference together with a disparity in electrical properties between the two regions results in electrical (Maxwell) stresses at the interface. We show via lubrication theory and via direct numerical simulations that the ring effect typically observed may be suppressed or augmented via appropriate use of electric fields. EPSRC DTG

  2. Immunohistochemical changes of maxillary sinus mucosa with underlying adenomatous polyposis

    Directory of Open Access Journals (Sweden)

    Yu. M. Andreychyn

    2017-04-01

    Full Text Available In maxillary sinusitis pathogenesis researchers paid much attention to the role of paranasal sinuses, sinus drainage dysfunctions, mucocyliary transport system disorders in cases of damage of sinuses mucous membranes. Pathohistological, immunohistochemical and morphometric studies of mucosa were conducted. However, to evaluate the local inflammatory response in cases of chronic sinusitis and improvement of individual therapy for the patients, the immunohistochemical studies of the composition of inflammatory immunocyte infiltrates of paranasal sinus mucosa and adenoid vegetations bioptate would be reasonable. The aim of the research was to study the immunohistochemical content of immunocyte infiltrates of paranasal sinuses adenomatous polyps in biopsy samples of patients suffering from chronic sinusitis. Materials and Methods. Paranasal sinuses adenomatous polyps removed during surgery of 22 patients suffering from chronic maxillary sinusitis were studied. Immunohistochemical studies of paraffin sections of the removed polyps were performed using appropriate primary antibodies and visualization systems DAKO EnVision+ System (DAKO, Denmark. Results. During microscopic investigation of paranasal sinuses adenomatous polyps of the patients suffering from chronic sinusitis, moderate and severe immunocyte infiltration of polyp stroma sometimes with focal follicular structures was presented. Immunohistochemical investigation of the cellular composition of these infiltrates proved the presence of CD8+ T-killer cells, CD4+ T-helper cells, CD68+ macrophages as well as single plasma cells. The irregularity of these cells markers expression was determined in dependence of the severity of inflammatory cell infiltration. Conclusions. In the patients with chronic maxillary polyp sinusitis with mild and moderate immunocyte infiltration of paranasal sinus mucosa, among immune cells the CD4+ T-lymphocytes occupied the most relative area in polyp stroma, plasma

  3. Enamel susceptibility to red wine staining after 35% hydrogen peroxide bleaching

    Directory of Open Access Journals (Sweden)

    Sandrine Bittencourt Berger

    2008-06-01

    Full Text Available Concern has been expressed regarding the staining of enamel surface by different beverages after bleaching. This study investigated the influence of 35% hydrogen peroxide bleaching agents on enamel surface stained with wine after whitening treatments. Flat and polished bovine enamel surfaces were submitted to two commercially available 35% hydrogen peroxide bleaching agents or kept in 100% humidity, as a control group (n = 10. Specimens of all groups were immersed in red wine for 48 h at 37°C, immediately, 24 h or 1 week after treatments. All specimens were ground into powder and prepared for the spectrophotometric analysis. Data were subjected to two-way analysis of variance and Fisher's PLSD test at 5% significance level. The amount of wine pigments uptake by enamel submitted to bleaching treatments was statistically higher than that of control group, independently of the evaluation time. Results suggested that wine staining susceptibility was increased by bleaching treatments.

  4. Deoxyribonuclease I (DNase I) typing from semen stains: low enzyme activity in vaginal fluids does not interfere with seminal DNase I typing from mixture stains.

    Science.gov (United States)

    Sawazaki, K; Yasuda, T; Nadano, D; Iida, R; Takeshita, H; Uchide, K; Kishi, K

    1993-09-01

    We describe the use of deoxyribonuclease I (DNase I) polymorphism for individualization of semen in body fluid stain mixtures, as a means of providing new and more useful information to practicing forensic biologists as a genetic marker. We have already reported that human DNase I isozyme patterns from different subjects are classifiable into ten groups. Isoelectric focusing of DNase I isozymes on polyacrylamide gel (IEF-PAGE, pH 3.5 to 5) was accomplished using a 0.5 mm thick gel. Pretreatment of semen samples with neuraminidase enhanced the isozyme band resolution and sensitivity. Activity detection using the dried agarose film overlay (DAFO) procedure was reliable, sensitive and simple, with high resolution, and the phenotypes of DNase I were determined in semen stains of about 0.3 microL stored at room temperature for up to a year in most of the samples tested. The DNase I types in semen stains were correlated with the types found in the corresponding blood and urine samples, although most of the vaginal fluid samples had no typable DNase I activity. This is considerably advantageous for seminal individualization from body fluid mixture stains in criminal cases. An evaluation of DNase I typing by IEF-PAGE and DAFO was also performed on casework samples submitted to our laboratory, and the results showed that DNase I was expected to be one of the most useful individualization marker of semen in practical application.

  5. An automated double staining procedure for bone and cartilage.

    Science.gov (United States)

    Miller, D M; Tarpley, J

    1996-03-01

    Differential skeletal staining is an important part of developmental toxicologic studies. Traditionally these studies have required time-consuming differentiation of one or both stains used and careful attention to the maceration step to prevent specimen destruction. We present a fully automated protocol which does not require differentiation of either dye and incorporates a controlled maceration step which is highly reproducible. This has resulted in high quality staining that is reproducible, stable, and can be done in volume with minimal personnel time. The process involves the staining of skinned, eviscerated specimens fixed in 95% ethanol. Using an automated tissue processor, the specimen is stained in alcian blue for 24 hr, macerated in 3% potassium hydroxide for 24 hr and stained with murexide for 24 hr. The specimens are cleared and preserved in glycerol. Within three days specimens have red stained bone and blue stained cartilage. The procedure was developed using 20-day-old Sprague-Dawley rat fetuses to evaluate the feasibility of using the procedure for teratology studies involving the fetal skeleton. Evenly stained specimens can be examined within three days and stored for years without loss of staining.

  6. New Methylene Blue Stain for Malaria Detection on Thin Smears

    Directory of Open Access Journals (Sweden)

    Himanshu D. Mulay

    2017-01-01

    Full Text Available Background: Malaria is the most important parasitic infection of man. Microscopy remains gold standard in malaria diagnosis. Management of malaria requires rapid detection of parasite in human blood. Hence there is a need to develop another diagnostic method with less limitation, which will address this issue. Aim and Objectives: To find a low cost reliable and accurate method for malaria detection on peripheral smear. Material and Methods: A prospective study of 40 cases was done. Two thin smears were prepared for each case; one was stained with Leishman stain and other with new methylene blue stain and examined under oil immersion. The smears were examined individually by two pathologists and results were prepared. Different parasitic morphologic forms were looked for. Parasitemia percentage was calculated. We also compared number of fields required to diagnose with both stains in positive cases. Results: In this study we found that 25 (83.3% cases were detected in less than 50 fields using New Methylene blue stain against 18 (60.0% cases with Leishman stain. We also found 100% sensitivity and specificity for New Methylene blue stain, whereas Leishman stain showed 90% sensitivity and specificity of 85%. Conclusion: The detection of malaria parasite was considerably easy with New Methylene Blue stain and required less time in comparison with Leishman stain.

  7. [Application of histochemical staining in diagnosis of osteosarcomas].

    Science.gov (United States)

    Li, Qing; Gong, Xi-qi; Ma, Fu-cheng; Zhao, Yi-ling; Zhu, Xiao-hui

    2005-08-01

    To study the histochemical staining in the diagnosis of osteosarcoma. To compare the effectiveness of picrosirius red, improved Ponceau trichrome and Masson trichrome staining methods on bone formation tissues in conventional osteosarcoma, paraosteal osteosarcoma, periosteal osteosarcoma, extraskeletal osteosarcoma, inflammatory myofibroblastic tumour, malignant fibrohistiocytoma, chondrosarcoma, fibrosis with ossification and calcification. With modified Ponceau trichrome staining, bone formation tissues showed a homogenous, orange-red interblended with blue in color. From osteoid to mature bone the color changed from orange-red, light blue to dark blue. Fibrotic tissue was stained blue in color with striated appearance. Cartilage was not stained. Picrosirius red method gave bone formation tissues homogenous staining. Along with bone maturation, from osteoid tissue to mineralized bones, the color showed changes from light red, yellow, orange-red, red to dark purple. The cartilage demonstrated homogenous light red in color. Fibrous tissue stained red interblended with yellow in color, striated in shape. With Masson trichrome staining osteoid displayed pale blue and mineralized bone showed dark blue in color. Fibrotic tissue showed a striated blue staining. The modified Ponceau trichrome and Picrosirius red staining methods are better than Masson trichrome to demonstrate bone formation tissue in osteosarcoma. The former two methods could be also used in study on bone formation.

  8. Effect of bleaching on staining susceptibility of resin composite restorative materials.

    Science.gov (United States)

    Celik, Ciğdem; Yüzügüllü, Bulem; Erkut, Selim; Yazici, A Rüya

    2009-01-01

    Effect of bleaching procedures on staining susceptibility of resin restorative materials is still questionable. The aim of this study was to evaluate the staining susceptibility of restorative materials bleached with 20% carbamide peroxide home bleaching agent and subsequently immersed in coffee and tea. Forty-two disk-shaped specimens were fabricated for each of the resin composites (Filtek Supreme XT [3M ESPE, St. Paul, MN, USA], Ceram-X Mono [Dentsply, Konstanz, Germany], and Aelite All Purpose Body [BISCO, Inc., Shaumburg, IL, USA]). The baseline color values were measured with a spectrophotometer. The specimens of each restorative material were randomly divided into two groups (N = 21). While the first group specimens were stored in distilled water (nonbleaching group-control), bleaching agent (Opalescence PF 20% [Ultradent Poducts, South Jordan, UT, USA]) was applied on the top surface of each specimen of the second group (bleaching group). After color change values were measured, the specimens were randomly divided into three subgroups (N = 7) according to the staining solutions. The color change values (DeltaE*ab) were calculated and the data were subjected to analysis of variance. Statistical significance was declared if the p value was 0.05 or less. There was no statistically significant difference within each restorative material's DeltaE*ab values after bleaching (p = 0.714). Also, the staining solutions did not cause a statistically significant difference between DeltaE*ab values of bleaching compared with nonbleaching groups (p = 0.146). Significant interaction was found only between restorative materials and staining solutions (p = 0.000). Bleaching of the tested resin composites did not increase their susceptibility to extrinsic staining in vitro. CLINICAL SIGNIFICANCE Bleaching did not affect staining susceptibility of the tested resin composite restorative materials. (J Esthet Restor Dent 21:407-415, 2009).

  9. Impact and mechanistic role of oral contraceptive pills on the number and epithelial type of ovarian cortical inclusion cysts; a clinicopathology and immunohistochemical study.

    Science.gov (United States)

    DastranjTabrizi, Ali; MostafaGharabaghi, Parvin; SheikhzadehHesari, Farzam; Sadeghi, Liela; Zamanvandi, Sharareh; Sarbakhsh, Parvin; Ghojazadeh, Morteza

    2016-03-22

    Ovarian epithelial cancers are among the most lethal women's cancers. There is no doubt about the preventive role of oral contraceptive pills (OCPs) in development of ovarian cancers. But, there are limited numbers of studies to address the effect of these agents on the number of cortical inclusion cysts (CICs), their epithelial type and suppression of the metaplastic phenomenon by these pills. The aim of this study was to clarify the role of these agents in the prevention of these cyst formation and tubal metaplasia and also examine the mesenchymal-epithelial transition theory in this context by immunohistochemical methods. The representative section(s) of ovarian cortex from a total number of 201 consecutive total abdominal hysterectomy with bilateral or unilateral salpingo-oophorectomy specimens were examined for mean number of CICs and their epithelial type between two groups of the patients. Group A included the patients who were on oral contraceptive pills for more than 5 years. All of the subjects with other contraceptive methods or a history of less than 5 years contraceptive pills usage were stratified in group B. Sections from 20 cases in which more than five inclusion cysts were found, were selected for IHC staining with calretinine and PAX8 as markers for mesothelium and mullerian epithelium respectively. The mean age of the patients was 51.67 years with no significant differences between two groups. The mean number of cysts were 1.27 and 3.23 in group A and B respectively (P =0.0001). Similarly the mean number of CICs, lined by tubal epithelium, was significantly different between two groups (0.65 vs 2.65, P =0.0001). In IHC staining 123 out of 150 CICs (82 %) were PAX+ while only 7 CICs (4.8 %) showed positive reaction for calretinin irrespective of type of epithelium. Our findings showed that the use of OCP for more than five years in women, significantly prevents development of cortical inclusion cysts in the ovaries which lined by tubal

  10. Comparative analysis of two and three marker immunohistochemical cocktails in the diagnosis of prostata carcinoma

    DEFF Research Database (Denmark)

    Dabir, Parag Deepak; Ottosen, Peter; Hamilton-Dutoit, Stephen Jacques

    2012-01-01

    Comparative analysis of two and three marker immunohistochemical cocktails in the diagnosis of prostata carcinoma......Comparative analysis of two and three marker immunohistochemical cocktails in the diagnosis of prostata carcinoma...

  11. Immunohistochemical Expression of Leptin (Ob-protein) in Experimentally Hypertensive Rat Kidney Tissues

    National Research Council Canada - National Science Library

    Fikret Gevrek; Meral Oncu; Kanat Gulle; Dilek Bayram; Erdal Karaoz

    2016-01-01

    Objective: Leptin is an ob gene protein which has a 16 kDa weight. The aim of this study is to determine immunohistochemically the correlation between immunohistochemical expression of Leptin and hypertension in rat kidney tissues...

  12. Validation of anti-FXR1 antibodies in the canine species and application to an immunohistochemical study of canine oral melanomas

    Directory of Open Access Journals (Sweden)

    Laura Nordio

    2017-05-01

    Full Text Available FXR1 (Fragile X mental retardation-related protein 1 is a cytoplasmic RNA binding protein, which genetic expression has been related to metastatic potential in human melanoma. The aims of the present study were: the validation of two commercially available clones of polyclonal anti-human FXR1 antibody in dogs; their application to investigate FXR1 expression in a group of canine oral melanomas. Anti-FXR1 antibody was not previously validated in the canine species. Two different commercially available polyclonal anti-FXR1 antibodies (respectively made in goat and in rabbit were used. FXR1 protein in canine serum was identified by western blot after SDS-PAGE, using human serum as control. FXR1 immunohistochemical expression was tested in a series of normal tissues, that are expected to express FXR1, and in 31 cases of oral melanomas. The final immunohistochemical protocol used heat-induced unmasking and overnight incubation. FXR1 protein bands in canine serum were detected by tested antibodies, in a more specific way by the rabbit antibody. FXR1 immunohistochemical staining was positive in all tested organs, with different levels of expression. FXR1 was also expressed in 31/31 tested melanomas, with variable intensity and percentage of positive cells (Figure 1. Equal results were achieved with the two antibodies in 8 cases of melanoma, whereas there were variable differences in 22, and one case stained only with goat antibody. The rabbit antibody gave less background staining. This study validated anti-FXR1 antibodies for use in the canine species. This protein was expressed in various normal tissues, as well as in the tested neoplasms. Significance of different level of expression is undergoing evaluation with further studies.

  13. A pre-application drop containing carboxymethylcellulose can reduce multipurpose solution-induced corneal staining.

    Science.gov (United States)

    Paugh, Jerry R; Marsden, Harue J; Edrington, Timothy B; Deland, Paul N; Simmons, Peter A; Vehige, Joseph G

    2007-01-01

    Use of polyhexanide based multipurpose solutions (MPSs) for contact lens disinfection has been linked to low-grade corneal staining. In vitro data suggest that carboxymethylcellulose (CMC) may neutralize polyhexanides. The purpose of this investigation was to determine whether a pre-application drop of CMC reduces polyhexanide staining in vivo. Thirty adapted soft contact lens (SCL) wearers participated in this investigator-masked, randomized, two-way cross-over study. Subjects wore a new Group II lens (alphafilcon A, 66% water) daily for 4 weeks and disinfected lenses using a MPS containing 0.0001% polyaminopropyl biguanide. A lens lubricant containing either CMC or povidone as the primary viscolyzer was applied to the lens each day before lens wear. Biomicroscopic signs and symptomatology were assessed. The difference in scores, 0 to 4 weeks and the difference between lubricants were analyzed. The cumulative fluorescein staining scores for combined eyes demonstrated a significant increase over time (e.g., cumulative staining score; p=0.004 and ppolyhexanide MPS. This result is consistent with a proposed mechanism for CMC to neutralize cationic disinfectants and may offer clinicians another means to reduce this type of corneal staining.

  14. Lugol staining pattern in background epithelium of patients with esophageal squamous cell carcinoma.

    Science.gov (United States)

    Katagiri, Atsushi; Kaneko, Kazuhiro; Konishi, Kazuo; Ito, Hiroaki; Kushima, Miki; Mitamura, Keiji

    2004-01-01

    Squamous cell carcinoma of the esophagus often arises in the setting of chronic esophagitis. We investigated whether chronic esophagitis was associated with carcinogenesis in the esophageal squamous epithelium. Videoendoscopy with Lugol staining was performed in 70 patients with invasive carcinoma of the esophagus. We especially focused the study on background epithelium of the esophagus, then background epithelium was classified into two groups according to differences in Lugol staining patterns. Following Lugol solution spraying, background epithelium showing uniform greenish-brown staining was defined as having a uniform pattern. In contrast, when multiple Lugol-unstained speckles were present throughout the esophagus, the pattern was defined as speckled. Furthermore, we also investigated whether glycogenic acanthosis is present or not in background epithelium. Chronic esophagitis was present in 11 of 70 patients (16%) with invasive carcinoma, indicating a speckled pattern in background epithelium following Lugol solution spraying. The remaining 84% of patients with invasive carcinoma showed normally uniform Lugol staining background epithelium. Glycogenic acanthosis was found in 65 (93%) of 70 patients. Approximately 80% of patients with esophageal squamous cell carcinoma showed normal Lugol staining of background epithelium. Field carcinogenesis is postulated to be not predominant in the development of esophageal squamous cell carcinoma in our Japanese subjects. In contrast, glycogenic acanthosis of the esophagus was associated with the background epithelium accompanied with esophageal squamous cell carcinoma.

  15. Biochemical and immunohistochemical estrogen and progesterone receptors in adenomatous hyperplasia and endometrial carcinoma: correlations with stage and other clinicopathologic features

    DEFF Research Database (Denmark)

    Nyholm, H C; Nielsen, A L; Lyndrup, J

    1992-01-01

    and immunohistochemical analysis were used. The immunohistochemical analysis receptor content was estimated semiquantitatively by a total and a cancer immunohistochemical histologic score. Multiple regression analysis was used in testing independence of established correlations. RESULTS: Estrogen and progesterone...

  16. Comparative analysis of cell proliferation ratio in plaque and erosive oral lichen planus: An immunohistochemical study

    Directory of Open Access Journals (Sweden)

    C Pramod Redder

    2014-01-01

    Full Text Available Background: Proliferating cell nuclear antigen (PCNA is a nuclear protein synthesized in the late G1 and S-phase of the cell cycle. Detection of this protein represents a useful marker of the proliferation status of lesions. This study has been carried out to evaluate the cell proliferation rate in oral lichen planus (OLP and comparison between plaque and erosive lichen planus, which indicates the potential for malignant transformation. Materials and Methods: This study was comprised of 64 cases of histologically proven lichen planus, out of which 32 cases of plaque and erosive each was taken. Two sections were taken from each, one for H and E staining to verify histological diagnosis according to Eisenberg criteria, other sections were stained according to super sensitive polymer horse radish peroxidise method for identifying immunohistochemical expression of PCNA. Data were statistically analyzed by Tukey high-range statistical domain test. Statistically significant P value was considered <0.05. Results: In two types of lichen planus, erosive type (66.86% showed higher expression of PCNA followed by plaque (17.07%. Overall, P value was <0.001, which was statistically significant. It indicates that proliferation activity is more in erosive lichen planus followed by plaque type, which ultimately results in increased rate of malignant transformation. Conclusion: PCNA is a good nuclear protein marker to evaluate the proliferation status of OLP. Out of the two types of lichen planus, erosive type possesses more proliferative ratio and chances of malignant change is more in this type. It emphasizes the importance of long-term follow-up with erosive type when compared with plaque type.

  17. Immuno-Histochemical and Quantitative Study of Melanocytes and Melanin Granules in Oral Epithelial Dysplasia.

    Science.gov (United States)

    Honwad, Swapna; Ravi, S V; Donoghue, Mandana; Joshi, Manjiri

    2017-07-01

    Oral Epithelial dysplasia (OED) is a potentially malignant disorder that is characterized by the presence of architectural and cytological changes. One of the prime factors responsible for the development of these lesions is the usage of tobacco. A variety of factors provide protective mechanism in order to prevent the effects of chemotoxic agents including tobacco products of which, melanin pigmentation is one of the vital elements. Role of melanocytes in progression of OED has remained unclear, so the present study was done to evaluate density of melanocyte and melanin granules in different grades of epithelial dysplasia and to correlate both findings with different grades of epithelial dysplasia. The study included 60 OED cases, of which three histopathogical sections were prepared from each block. The sections were stained with Hematoxylin and Eosin, Masson Fontana and Human Melanoma Black (HMB-45), an immunohistochemical stain. Quantification of melanin granules was evaluated under 40X magnification using arbitrary scale with micrometer square as, 0= Absence of melanin granules, 1= Rare and scattered melanin granules, 2= Dense but not aggregated melanin granules, 3= Dense and aggregated melanin granules. Density of melanocytes was evaluated under 10X magnification. Five consecutive fields were evaluated for melanocytes and melanin granules starting from the field of highest density. There was an insignificant increase in number of melanocytes and melanin granules in mild and moderate dysplasia compared to normal but significant reduction was observed in severe dysplasia. The decrease in number of melanocytes and melanin granules was proportional to severity of epithelial dysplasia. This could be due to chronic irritation by chemical products leading to death of melanocytes.

  18. Factors influencing extract of Hibiscus sabdariffa staining of rat testes.

    Science.gov (United States)

    Bassey, R B; Bakare, A A; Peter, A I; Oremosu, A A; Osinubi, A A

    2012-08-01

    Some plant extracts can be used in biology and medicine to reveal or identify cellular components and tissues. We investigated the effects of time and concentration on staining of histological sections of rat testes by an acidified extract of Hibiscus sabdariffa. An ethanolic extract of H. sabdariffa was diluted using 1% acetic acid in 70% ethanol to stain histological sections of testes at concentrations of 0.2, 0.1 and 0.05 g/ml for 5, 10, 15, 30, 45 and 60 min. The sections of testes were stained deep red. The staining efficiency of H. sabdariffa was greater at a high concentration and required less time to achieve optimal staining. H. sabdariffa is a strongly basic dye that can be used for various diagnostic purposes. Staining time and concentration must be considered to achieve optimal results.

  19. A useful single-solution polychrome stain for plant material...Brook Cyte-Chrome I.

    Science.gov (United States)

    Stanley L Krugman; Julia F. Littlefield

    1968-01-01

    Fresh and chemically fixed sectioned plant material can be quickly stained by applying a Brook Cyte Chrome I polychrome stain. Staining time averaged only about 10 minutes. And exact timing of staining and de-staining is not as critical as with most of the commonly used stains. The overall quality is comparable to that of the traditional stains.

  20. Near-UV laser treatment of extrinsic dental enamel stains.

    Science.gov (United States)

    Schoenly, J E; Seka, W; Featherstone, J D B; Rechmann, P

    2012-04-01

    The selective ablation of extrinsic dental enamel stains using a 400-nm laser is evaluated at several fluences for completely removing stains with minimal damage to the underlying enamel. A frequency-doubled Ti:sapphire laser (400-nm wavelength, 60-nanosecond pulse duration, 10-Hz repetition rate) was used to treat 10 extracted human teeth with extrinsic enamel staining. Each tooth was irradiated perpendicular to the surface in a back-and-forth motion over a 1-mm length using an ∼300-µm-diam 10th-order super-Gaussian beam with fluences ranging from 0.8 to 6.4 J/cm(2) . Laser triangulation determined stain depth and volume removed by measuring 3D surface images before and after irradiation. Scanning electron microscopy evaluated the surface roughness of enamel following stain removal. Fluorescence spectroscopy measured spectra of unbleached and photobleached stains in the spectral range of 600-800 nm. Extrinsic enamel stains are removed with laser fluences between 0.8 and 6.4 J/cm(2) . Stains removed on sound enamel leave behind a smooth enamel surface. Stain removal in areas with signs of earlier cariogenic acid attacks resulted in isolated and randomly located laser-induced, 50-µm-diam enamel pits. These pits contain 0.5-µm diam, smooth craters indicative of heat transfer from the stain to the enamel and subsequent melting and water droplet ejection. Ablation stalling of enamel stains is typically observed at low fluences (Laser ablation of extrinsic enamel stains at 400 nm is observed to be most efficient above 3 J/cm(2) with minimal damage to the underlying enamel. Unsound underlying enamel is also observed to be selectively removed after irradiation. Copyright © 2012 Wiley Periodicals, Inc.

  1. Diagnosing periprosthetic infection: false-positive intraoperative Gram stains.

    Science.gov (United States)

    Oethinger, Margret; Warner, Debra K; Schindler, Susan A; Kobayashi, Hideo; Bauer, Thomas W

    2011-04-01

    Intraoperative Gram stains have a reported low sensitivity but high specificity when used to help diagnose periprosthetic infections. In early 2008, we recognized an unexpectedly high frequency of apparent false-positive Gram stains from revision arthroplasties. The purpose of this report is to describe the cause of these false-positive test results. We calculated the sensitivity and specificity of all intraoperative Gram stains submitted from revision arthroplasty cases during a 3-month interval using microbiologic cultures of the same samples as the gold standard. Methods of specimen harvesting, handling, transport, distribution, specimen processing including tissue grinding/macerating, Gram staining, and interpretation were studied. After a test modification, results of specimens were prospectively collected for a second 3-month interval, and the sensitivity and specificity of intraoperative Gram stains were calculated. The retrospective review of 269 Gram stains submitted from revision arthroplasties indicated historic sensitivity and specificity values of 23% and 92%, respectively. Systematic analysis of all steps of the procedure identified Gram-stained but nonviable bacteria in commercial broth reagents used as diluents for maceration of periprosthetic membranes before Gram staining and culture. Polymerase chain reaction and sequencing showed mixed bacterial DNA. Evaluation of 390 specimens after initiating standardized Millipore filtering of diluent fluid revealed a reduced number of positive Gram stains, yielding 9% sensitivity and 99% specificity. Clusters of false-positive Gram stains have been reported in other clinical conditions. They are apparently rare related to diagnosing periprosthetic infections but have severe consequences if used to guide treatment. Even occasional false-positive Gram stains should prompt review of laboratory methods. Our observations implicate dead bacteria in microbiologic reagents as potential sources of false-positive Gram

  2. Lasers or light sources for treating port-wine stains

    DEFF Research Database (Denmark)

    Faurschou, Annesofie; Olesen, Anne Braae; Leonardi-Bee, Jo

    2011-01-01

    Port-wine stains are birthmarks caused by malformations of blood vessels in the skin. Port-wine stains manifest themselves in infancy as a flat, red mark and do not regress spontaneously but may, if untreated, become darker and thicker in adult life. The profusion of various lasers and light...... sources makes it difficult to decide which equipment is the best for treating port-wine stains....

  3. Immunohistochemical localization of glutathione s-transferase isoenzymes (gsta, Gstp, Gstm4, And Gstt1) and tumour marker p53 in matched tissue from normal larynx and laryngeal carcinoma: correlations with prognostic factors.

    Science.gov (United States)

    Sedat, Aydin; Serpil, Oguztuzun; Nurdan, Gurbuz; Aylin, Gul; Arif, Sanli; Muzeyyen, Ozhavzali; Bulent, Satar; Sezen, Ozan; Nimet, Karadayi

    2010-10-01

    The immunohistochemical staining characteristics of glutathione S-transferase (GST) alpha (GSTA), pi (GSTP), mu (GSTM4), and theta (GSTT1) and P53 were investigated in laryngeal squamous cell carcinoma (LSCC) cases and normal laryngeal tissue from 46 patients. The relationships between expression of the GST isoenzymes and some clinicopathologic features were also examined. For immunohistochemical studies, tissues from 46 patients with LSCC at the Dr. Lütfi Kirdar Kartal Education and Research Hospital were used. The relationship between expression of the GST isoenzymes and P53 in normal and tumour tissue was analyzed using the Wilcoxon signed rank test. The correlation between GST isoenzymes and P53 and clinicopathologic data was also examined using the Spearman rank test. When the normal and tumour tissues of these cases were compared according to their staining intensity and percentage of positive staining, GSTA expression in normal cells was significantly higher than in tumour cells, and GSTP and P53 expression was higher in tumour cells (p .05). There was no correlation between P53 and GST expression in patients with LSCC. When the immunohistochemical results of GST isoenzymes and P53 were correlated with the clinical parameters, GSTA expression was increased in poorly differentiated laryngeal tumour, but GSTM4 and GSTT1 expression was decreased (p < .05). According to these results, GST-A, -P, and T1 and P53 were important in the diagnosis of LSCC.

  4. Utility of immunohistochemical analysis of KAI1, epithelial-specific antigen, and epithelial-related antigen for distinction of chromophobe renal cell carcinoma, an eosinophilic variant from renal oncocytoma.

    Science.gov (United States)

    Ohe, Chisato; Kuroda, Naoto; Takasu, Kosho; Senzaki, Hideto; Shikata, Nobuaki; Yamaguchi, Tadanori; Miyasaka, Chika; Nakano, Yorika; Sakaida, Noriko; Uemura, Yoshiko

    2012-06-01

    Distinction of renal oncocytoma (RO) from chromophobe renal cell carcinoma (ChRCC) is important because their clinical behavior is different. As part of a search for the best available immunohistochemical markers to distinguish ChRCC from RO, we investigated the immunohistochemical profiles of these tumors. We selected 30 renal tumors consisting of ChRCC, typical variant (n = 14), ChRCC, eosinophilic variant (n = 6), and RO (n = 10). Their expression of cytokeratin (CK) 7, KAI1, epithelial-specific antigen (ESA), epithelial-related antigen (ERA), Claudin- 7, and Claudin-8 was studied using an autostainer. Immunoreactivity was assessed based on a combined score of the extent and intensity of staining. Compared to RO, a significantly higher percentage of the total ChRCCs stained positive for CK7 (85% vs. 10%, respectively), KAI1 (90% vs. 10%), ESA (95% vs. 10%), ERA (95% vs. 10%), and Claudin-7 (95% vs. 20%) (P < 0.001). Additionally, there was a significant difference between the percentage of ChRCC eosinophilic variant (ChRCC-E) and RO that stained positive for KAI1 (100% vs. 10%, respectively), ESA (83% vs. 10%), and ERA (83% vs. 10%) (P < 0.001). We recommend immunohistochemical analysis of KAI1, ESA, and ERA to distinguish ChRCC-E from RO.

  5. Mitotic activity of multinucleated giant cells with glial fibrillary acidic protein immunoreactivity in glioblastomas: an immunohistochemical double labeling study.

    Science.gov (United States)

    Takeuchi, Hiroaki; Sato, Kazufumi; Ido, Kazunori; Kubota, Toshihiko

    2006-05-01

    To investigate the mitotic activity of multinucleated giant cells (MNGCs) with glial fibrillary acidic protein (GFAP) in glioblastomas, double immunohistochemical staining for GFAP and Ki67 was performed in formalin-fixed and paraffin-embedded specimens obtained from 12 primary glioblastomas with MNGCs including three giant cell glioblastomas. The Ki67 labeling index (LI:%) of GFAP+ tumor cells ranged from 0 to 5.6 (2.5+/-1.7, mean+/-standard deviation). The Ki67 LI of GFAP- tumor cells ranged from 18.6 to 35.9 (24.7+/-6.6). The Ki67 LI of GFAP+ cells was significantly lower than that of GFAP- cells (Pglioblastomas. MNGCs identified in glioblastomas may develop via not only the proliferation of abnormal nuclei in a single tumor cell but also other processes.

  6. Characteristic Ber-EP4 and EMA expression in sebaceoma is immunohistochemically distinct from basal cell carcinoma.

    Science.gov (United States)

    Fan, Y S; Carr, R A; Sanders, D S A; Smith, A P; Lazar, A J F; Calonje, E

    2007-07-01

    There is considerable overlap between the histological features of sebaceoma and basal cell carcinoma (BCC). The distinction between these two tumours is important due to the often more locally aggressive nature of BCC and the association of sebaceoma with the Muir-Torre syndrome. The aim of this study was to describe the immunohistochemical reactivity of the cells in sebaceoma to Ber-EP4 and epithelial membrane antigen (EMA) and investigate the utility of this panel to differentiate sebaceoma from basal cell carcinoma. Immunohistochemistry of 25 sebaceomas for Ber-EP4 and EMA revealed unequivocal negative expression of Ber-EP4 in 24 of 25 sebaceomas. A single case exhibited focal weak Ber-EP4 staining, predominantly in mature sebocytes and in Ber-EP4 in combination with EMA, both widely used immunomarkers in histopathology, is a helpful aid in distinguishing sebaceoma from nodular BCC.

  7. Alpha-defensin expression in the gastric tissue of children with Helicobacter pylori-associated chronic gastritis: an immunohistochemical study.

    Science.gov (United States)

    Soylu, Ozlem Bekem; Ozturk, Yesim; Ozer, Erdener

    2008-04-01

    In this study, we evaluated the expression of alpha-defensin and its correlation with histological criteria in children with and without Helicobacter pylori-associated gastritis. Forty-five children were included. Immunohistochemical staining was performed and the relationship between alpha-defensin immunoscoring and H. pylori status and histological criteria was evaluated. Expression of alpha-defensin was significantly higher in the H. pylori-positive group (P < 0.001) and it was significantly associated with higher grades of chronic inflammation and neutrophil density (P < 0.001 for both). Our data show that alpha-defensin expression is increased in H. pylori infection in childhood and is associated with inflammatory tissue damage.

  8. Histopathological evaluation of ocular microsporidiosis by different stains

    Directory of Open Access Journals (Sweden)

    Sharma Savitri

    2006-06-01

    Full Text Available Abstract Background There is limited data on comparing stains in the detection of microsporidia in corneal biopsies. Hence we wanted to evaluate various stains for their ability to detect microsporidia in corneal tissue sections. Methods Four cases diagnosed with microsporidiosis on Hematoxylin and Eosin and Periodic Acid Schiff's stained sections of the corneal button between January 2002 and December 2004, were included. Further sections were prospectively stained with calcofluor white, Gram, Giemsa, Masson's trichrome, acridine orange, Gomori's methenamine silver, Gram's chromotrope and modified acid fast stain. The stained sections were analyzed for the spore characteristics in terms of size, shape, color contrast, cell wall morphology, waist band in cytoplasm and ease of detection. Results All sections showed microsporidial spores as 3 – 5 μm, oval bodies. 1% acid fast, Gram's chromotrope and GMS stains provided a reliable diagnosis of microsporidia as diagnostic waist band could be identified and good contrast helped distinguish the spores from inflammatory debris. Conclusion Considering the ease of performance, cost effectiveness and rapidity of the technique, 1% acid fast stain and Gram's chromotrope stain are ideal for the detection of microsporidia.

  9. Black stain and dental caries in Filipino schoolchildren.

    Science.gov (United States)

    Heinrich-Weltzien, Roswitha; Monse, Bella; van Palenstein Helderman, Wim

    2009-04-01

    Black stain is defined as dark pigmented exogenous substance in lines or dots parallel to the gingival margin and firmly adherent to the enamel at the cervical third of the tooth crowns in the primary and permanent dentition. This study was conducted to assess the prevalence of black stain on teeth of Filipino children and to determine a possible association between black stain and caries levels. The study was designed to test the following hypotheses: (i) the prevalence of black stain does not differ between children from schools with oral health intervention programs and those from schools without an intervention program, (ii) the prevalence of black stain does not differ in children attending easily accessible and remote schools, (iii) caries prevalence and caries experience do not differ in children with and without black stain and (iv) the caries distribution at the surface level does not differ in children with and without black stain. In total, 32 elementary schools were included. 19 schools with a comprehensive school-based preventive oral health program, seven schools with a basic preventive program and six control schools. All sixth graders of these schools (n=1748) aged 11.7+/-1.1 years were clinically examined for black stain. DMFT was assessed in 1121 children by seven calibrated dentists using WHO criteria. DMFS was scored in 627 children by two calibrated dentists. Black stain was found in 16% of this population. The prevalence of black stain did not differ significantly between children attending schools with different oral health intervention programs. Thus, hypothesis 1 was accepted. The prevalence of black stain was significantly higher (Pcaries prevalence and caries experience than children without black stain. Thus, hypothesis 3 was rejected. No difference was found in the DMFS pattern of occlusal, smooth and proximal surfaces between children with and without black stain. Thus hypothesis 4 was accepted. The presence of black stain is

  10. Epithelial ovarian tumors: Clinicopathological correlation and immunohistochemical study

    Directory of Open Access Journals (Sweden)

    Pooja S Naik

    2015-01-01

    Full Text Available Background: Ovarian cancer is the third leading site of cancer among women, trailing behind cervix and breast cancer. Aim: This study was undertaken to analyze the immunohistochemical (IHC profile of estrogen receptors (ER, progesterone receptors (PR, Ki-67, and p53 in various ovarian epithelial tumors and attempt correlation with clinical and histopathological findings. Materials and Methods: The present study was conducted over a period of 4 years. A technique of manual tissue array was employed for cases subjected for IHC. The primary antibodies used were ER, PR, p53, and Ki-67. A correlation was attempted between histopathological and IHC findings. Results were subjected to statistical analysis. Software program "the primer of biostatistics 5.0" was used for calculation of interrelationships between the analyzed ER, PR, p53, and Ki-67 expression and histological factors by Pearson′s Chi-square test. The results were considered to be significant when the P < 0.05. Results: There were 110 cases of surface epithelial ovarian tumors (SEOT encountered over the period of 4 years. The expression of ER was more in malignant tumors (13/16, 81.25% than borderline (9/12, 75% and benign (20/82, 24.39%. As compared to ER, the expression of PR was more in benign (51/82, 62.19% than borderline (8/12, 66.67% and malignant tumors (9/16, 56.25%. The expression of PR was more in benign tumors than borderline and malignant tumors. However, this was not statistically significant (Chi-square = 0.335 with 2 degrees of freedom; P = 0.846. The expression of p53 was less in benign (5/82, 6.1% than borderline (9/12, 75% and malignant tumors (13/16, 81.25%. The expression of Ki-67 was more in malignant (4/82, 4.88% than borderline (10/12, 83.33% and benign tumors (15/16, 93.75%. In all the above cases, the difference was statistically significant (P < 0.05. There was statistically significant difference in the expression of ER, PR, p53, and Ki-67 in the patients with

  11. An induced extrinsic tooth stain prevention model to investigate whitening potential of sugar-free chewing gums.

    Science.gov (United States)

    Dodds, Michael Wj; Tian, Minmin; Ramirez, Lilian; Milleman, Jeffery L; Milleman, Kimberly R; Proskin, Howard M

    2017-12-01

    To establish an accelerated clinical test method to evaluate the effectiveness of sugar-free gums in prevention of the formation of extrinsic stains when chewed over a 2-week period in conjunction with daily tooth brushing. A secondary objective was to compare three methods for measuring extrinsic stain. 25 healthy adult volunteers were enrolled in a single center, examiner blind, randomized 4-way crossover clinical study. Starting with a stain-free baseline, subjects rinsed five times daily with freshly brewed black tea, followed either by chewing one of three different gums for 12 minutes or not chewing (negative control). Extrinsic stain was measured at 1 and 2 weeks by modified Lobene Stain Index (MLSI), digital imaging, and a Vita EasyShade spectrophotometer. At 2 weeks, MLSI scores showed a statistically significant mean reduction of 43% or greater versus no-gum control for all three gum treatments. Digital image analysis and Vita EasyShade measurement showed reductions of yellowness (measured by difference in ∆b* values between the three gums and the non-gum control treatment) ranging from 0.28 to 0.34 and 3.52 to 4.18 Δb* units, respectively, for subjects using the chewing gums versus no-gum control (Peffectively reduce new stain formation along with daily tooth brushing in as little as 2 weeks when used in conjunction with tea rinsing to help promote more rapid stain formation. All three test methods confirmed the results, albeit with different levels of statistical significance. A minor modification of gum base polymer, or change of flavors, did not significantly impact the prevention of new stain formation. Regular consumption of sugar-free chewing gum helps prevent extrinsic dental stain accumulation and provides a simple and enjoyable means for consumers to maintain their natural tooth color.

  12. Identification of risk factors of prostate adenocarcinoma recurrence after HIFU therapy using immunohistochemical markers

    Directory of Open Access Journals (Sweden)

    Popkov V.M.

    2014-12-01

    Full Text Available The purpose of this study was to identify risk factors for recurrence of prostate adenocarcinoma after HIFU therapy. Material and methods: Material for the study was obtained from patients diagnosed with adenocarcinoma before and after HIFU treatment. Morphological study was conducted using a standard staining, and immunohistochemical markers: PCNA, Amacr, E-cadherin, Bel2, Andr, Estr, VEGF, P53, PCNA. Results: After treatment in 89% of patients with initial prostate volume greater than 50 cc the signs of recurrence of adenocarcinoma were showed. At low risk for D'Amico after treatment the expression of proliferation markers, VEGF, Amacr significantly decreased. With a high degree of risk — increased expression of Bel2. After treatment the expression of the following markers: PCNA, Amacr, VEGF significantly increased in the group of patients with the presence of invasion. Conclusion: Patients with initial prostate volume less than 50 cc, low risk to D'Amico, the lack of perineural and perivascular invasion have a low risk of recurrence after HIFU therapy; patients at high risk for D'Amico, the presence of perineural and perivascular invasion initial and prostate volume greater than 50 cc, low-grade cribriform form of adenocarcinoma have a high risk of recurrence of adenocarcinoma. Recurrence of adenocarcinoma develops independently of the period after HIFU therapy.

  13. Immunohistochemical detection of autotaxin (ATX)/lysophospholipase D (lysoPLD) in submucosal invasive colorectal cancer.

    Science.gov (United States)

    Kazama, Shinsuke; Kitayama, Joji; Aoki, Junken; Mori, Ken; Nagawa, Hirokazu

    2011-12-01

    Autotaxin (ATX) is molecularly identical to lysophospholipase D (lysoPLD) and is a main enzyme producing lysophosphatidic acid (LPA), which mediates a broad range of cellular responses including stimulation of cell motility. Using immunohistochemical staining, we examined the expression of ATX/lysoPLD in 98 cases of early colorectal cancer with submucosal invasion. ATX/lysoPLD was highly expressed in infiltrating cells in tumor tissue in the submucosal layer, which were characterized as mast cells. The number of ATX/lysoPLD-positive cells was significantly greater in tumors with a macroscopically depressed lesion than in tumors without depression. The density of ATX/lysoPLD-positive cells tended to have a positive correlation with microvessel vascular density (MVD), while it was not correlated with vessel invasion and nodal metastases as well as lymphovascular vessel density (LVD). Our results suggest that local production of LPA through ATX/lysoPLD may weakly correlate with formation of a depressive lesion and tumor angiogenesis in the early stage of colorectal cancer.

  14. An immunohistochemical study of matrix proteins in the craniofacial cartilage in midterm human fetuses

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    S. Shibata

    2013-12-01

    Full Text Available Immunohistochemical localization of collagen types I, II, and X, aggrecan, versican, dentin matrix protein (DMP-1, martix extracellular phosphoprotein (MEPE were performed for Meckel’s cartilage, cranial base cartilage, and mandibular condylar cartilage in human midterm fetuses; staining patterns within the condylar cartilage were compared to those within other cartilaginous structures. Mandibular condylar cartilage contained aggrecan; it also had more type I collagen and a thicker hypertrophic cell layer than the other two types of cartilage; these three characteristics are similar to those of the secondary cartilage of rodents. MEPE immunoreactivity was first evident in the cartilage matrix of all types of cartilage in the human fetuses and in Meckel’s cartilage of mice and rats. MEPE immunoreactivity was enhanced in the deep layer of the hypertrophic cell layer and in the cartilaginous core of the bone trabeculae in the primary spongiosa. These results indicated that MEPE is a component of cartilage matrix and may be involved in cartilage mineralization. DMP-1 immunoreactivity first became evident in human bone lacunae walls and canaliculi; this pattern of expression was comparable to the pattern seen in rodents. In addition, chondroid bone was evident in the mandibular (glenoid fossa of the temporal bone, and it had aggrecan, collagen types I and X, MEPE, and DMP-1 immunoreactivity; these findings indicated that chondroid bone in this region has phenotypic expression indicative of both hypertrophic chondrocytes and osteocytes.

  15. CD98 immunoreactivity in multinucleated giant cells of glioblastomas: an immunohistochemical double labeling study.

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    Takeuchi, Hiroaki; Kubota, Toshihiko; Kitai, Ryuhei; Nakagawa, Takao; Hashimoto, Norichika

    2008-04-01

    CD98, which is identical to fusion regulatory protein-1 (FRP-1), has been reported to induce and regulate cell fusion and multinucleated giant cell formation. To investigate the association between CD98 and multinucleated giant cells (MNGCs) in glioblastomas, we investigate the CD98 immunoreactivity of MNGCs and the proliferative potential in CD98 immunoreactive MNGCs in paraffin-embedded sections obtained from patients with glioblastomas. Double immunohistochemical staining for CD98 and Ki67 as a mitotic marker were performed in formalin-fixed and paraffin-embedded specimens obtained from 16 patients with primary glioblastomas including MNGCs. Most CD98 immunoreactive (CD98+) tumor cells were negative for Ki67. CD98+ MNGCs were identified in 15 cases. CD98+ Ki67- MNGCs were identified in 14 cases and ranged in number from one to 48 (6.7 +/- 11.5). CD98- Ki67+ MNGCs were identified in 15 cases and ranged in number from one to 32 (11.1 +/- 9.6). Mitotic index (MI) of CD98+ MNGCs (4.8 +/- 2.7%) was significantly lower than that of CD98- MNGCs (91.1 +/- 24.6%) (P giant cell formation may be developed by fusion among CD98- producing cells in glioblastomas.

  16. Comparative immunohistochemical study of Bcl-X in ameloblastoma, keratocystic odontogenic tumor and adenomatoid odontogenic tumor

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    Shukla, Payal; Prabhu, Sudeendra; Jose, Maji; Sripathi Rao, B H

    2017-01-01

    Objectives: Since its recognition as a physiologic process associated with tumor, among molecular mechanisms involved in tumor progression, defects in regulation of apoptosis have generated an accelerating volume of research that has sought to elucidate the role of programed cell death in pathogenesis and treatment of various tumors. Therefore, this study was performed to understand better the diverse biological profile of epithelial odontogenic tumors with the help of immunohistochemical expression of Bcl-X protein. Materials and Methods: We studied Bcl-X protein expression in 45 cases of epithelial odontogenic tumors which included 15 cases each of ameloblastomas, keratocystic odontogenic tumor (KCOT) and adenomatoid odontogenic tumor (AOT) and correlated the expression with their growth pattern. Results: Cytoplasmic staining of Bcl-X revealed overexpression in ameloblastoma when compared to KCOT and AOT. Percentage of positive cells showed a statistically significant difference, P = 0.007 between ameloblastoma and KCOT, whereas P odontogenic tumors show diverse growth profiles. An increased Bcl-X expression was seen in ameloblastoma compared to KCOT and least expression in case of AOT which could be indicative of more aggressive biological behavior and increased cell survival activity of ameloblastoma than KCOT and AOT. This signifies the diagnostic relevance of this biomarker and also could be a possible regulator of the proliferative compartment by contributing in tumor progression and cytodifferentiation of epithelial odontogenic tumors. PMID:28479687

  17. A positive correlation between immunohistochemical expression of CD31 and mast cell tryptase in odontogenic tumors.

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    Kouhsoltani, Maryam; Halimi, Monireh; Dibazar, Sana

    2015-06-01

    In this study, we compared mast cell tryptase and CD31 expression between odontogenic tumors with the aim of predicting the clinical behavior of these lesions at the time of initial biopsy. We also evaluated the correlation between mast cell tryptase and CD31 expression to clarify the role of mast cells (MCs) in the growth of odontogenic tumors. Immunohistochemical staining with anti-MC tryptase and anti-CD31 antibodies was performed on 48 cases of odontogenic tumors including solid ameloblastoma (SAM), unicystic ameloblastoma (UAM), odontogenic myxoma (OM), cystic calcifying odontogenic tumor (CCOT) and adenomatoid odontogenic tumor (AOT). Ten high power fields were analyzed for each sample. Total MC count was significantly increased in SAM compared to other odontogenic tumors (podontogenic tumors (podontogenic tumors (p=0.018, r=0.34). Our findings suggest a role for MCs in aggressive clinical behavior of odontogenic tumors. The significant correlation found between MC count and microvessel density in odontogenic tumors is in agreement with the theory of participation of MCs in tumor progression. Targeting MC activity may represent an important nonsurgical therapeutic approach, especially for aggressive odontogenic tumors.

  18. Validation of an immunohistochemical assay for bovine cysticercosis, with comparison to a standard histological method.

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    Scandrett, W Brad; Haines, Deborah M; Parker, Sarah E; Robinson, Yves; Forbes, Lorry B; Brandt, Jef; Geerts, Stanny; Dorny, Pierre; Gajadhar, Alvin A

    2012-05-25

    The larval stage (syn Cysticercus bovis) of the human tapeworm Taenia saginata causes cysticercosis in cattle, which has both aesthetic and food safety implications to consumers of beef. A monoclonal antibody-based immunohistochemical (IHC) assay developed to improve postmortem diagnosis of this parasite and a standard histological method were assessed to determine their fitness for intended use. Sections from 169 known-positive specimens of T. saginata from experimentally or naturally infected cattle, and from 30 known-negative specimens and lesions of various etiologies from non-infected cattle, were tested. The IHC assay identified significantly more known positive bovine cysticerci than the histological method (91.7% and 38.5%, respectively). Positive IHC staining occurred on sections from other cestode species, but should not affect the diagnostic specificity of this assay for bovine cysticercosis, due to the different host and/or tissue preferences amongst these parasites. Use of the IHC assay should improve the reliability of diagnosing lesions caused by degenerated cysticerci, facilitating more effective and efficient control of bovine cysticercosis. Crown Copyright © 2011. Published by Elsevier B.V. All rights reserved.

  19. Phenotypic Investigation of Regenerated Epithelial Cells After Gonococcal Corneal Perforation: A Clinical, Histological, and Immunohistochemical Study.

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    Jongkhajornpong, Passara; Nakamura, Takahiro; Sotozono, Chie; Inatomi, Tsutomu; Kinoshita, Shigeru

    2015-11-01

    To determine the characteristics of regenerated epithelial cells after severe gonococcal infection after corneal perforation. Pathological tissue was obtained from the cornea at the time of surgery. Hematoxylin and eosin staining and immunohistochemical analysis were performed for cytoskeletal keratins (K12, K13, and K15), basement membrane and junctional markers (laminin 5, ZO-1 and Desmoplakin), and proliferative and mesenchymal markers (Ki67, α-SMA, and vimentin). A 42-year-old patient with severe gonococcal keratoconjunctivitis rapidly progressed to corneal perforation during administration of intensive topical and systemic antibiotics. After conservative treatment, the perforation healed and 5- × 3-mm corneal ectasia occurred with localized iris attachment. Complete closure of the cornea was confirmed by a negative Seidel test. After lamellar keratoplasty to improve corneal integrity and to prevent secondary glaucoma, the pathological tissue revealed a poorly organized epithelial layer at the regenerated ectatic area. The regenerated epithelial cells clearly expressed K12, ZO-1, and Desmoplakin with underlying laminin 5 (+) basement membrane. K15 and Ki67 expressions were observed predominantly at the limbal area but not in the regenerated area. α-SMA and vimentin were sporadically expressed in the underlying connective tissue. We speculate that the process of epithelial wound healing at the site of corneal perforation was responsible for migration of the surrounding epithelial cells. Although the regenerated cells expressed several cytokeratins and junctional markers, they remained disorganized and fragile.

  20. Microvasculature and healing potential of the inner versus outer preputial skin: preliminary immunohistochemical observations.

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    Elbakry, Adel; Matar, Adel; Zalata, Khaled; Zakaria, Ahmed; Al Atrash, Gamal

    2015-02-01

    To assess healing potential of the inner and the outer layers of the prepuce and to determine which layer is better for hypospadias neourethral reconstruction. The study has been carried out to assess the microvessels density and epidermal growth factor receptor (EGFR) concentration in the inner and the outer preputial layers. Specimens from the outer and the inner prepuce were harvested during hypospadias repair in 26 children. Control specimens were collected during elective circumcision of 10 normal and age-matched children. Sections were prepared, and immunohistochemical staining was done using monoclonal antibodies of CD34 (vascular marker) and of the EGFR. CD34-positive microvessels were assessed under the outer and the inner layers of the prepuce and were counted in five high-power fields under each layer. Expression of EGFR in both layers was assessed using H-score system. The density of microvessels and EGFR expression are significantly higher in control group either for inner or for outer preputial layers (p 0.05). In hypospadias patients, the healing potential of both inner and outer prepuce is nearly similar. However, it is markedly reduced than that of normal prepuce. It seems that both layers can be used for hypospadias repair without obvious preference to either of them. The usual tradition to use inner prepuce in hypospadias repair has no scientific evidence.

  1. Immunohistochemical and morphological features of a small bowel leiomyoma in a black crested macaque (Macaca nigra

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    Aristizabal-Arbelaez Mónica

    2012-06-01

    Full Text Available Abstract Background Spontaneous gastrointestinal neoplasms in non-human primates are commonly seen in aged individuals. Due to genetic similarities between human and non-human primates, scientists have shown increasing interest in terms of comparative oncology studies. Case presentation The present study is related to a case of an intestinal leiomyoma in a black crested macaque (Macaca nigra, kept on captivity by Matecaña Zoo, Pereira City, Colombia. The animal had abdominal distension, anorexia, vomiting, diarrhea and behavioral changes. Clinical examination showed an increased volume in the upper right abdominal quadrant caused by a neoplastic mass. The patient died during the surgical procedure. Necropsy revealed several small nodules in the peritoneum with adhesion to different portions of the small and large intestines, liver, stomach and diaphragm. Tissue samples were collected, routinely processed and stained by H&E. Microscopic examination revealed a mesenchymal tumor limited to tunica muscularis, resembling normal smooth muscle cells. Neoplastic cells were positive for alpha-smooth muscle actin and vimentin, and negative for cytokeratin AE1/AE3 by immunohistochemistry. Those morphological and immunohistochemical findings allowed to diagnose the intestinal leiomyoma referred above. Conclusion Neoplastic diseases in primates have multifaceted causes. Their manifestations are understudied, leading to a greater difficulty in detection and measurement of the real impact provides by this disease.

  2. Palpebro-orbital apocrine cystadenoma: immunohistochemical verification of a unique variant with a critical differential diagnosis.

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    Jakobiec, Frederick A; Bhat, Pooja; Kropp, Thomas M

    2010-01-01

    To describe a unique apocrine cystadenoma of the superonasal eyelid and anterior orbit. Clinical evaluation with axial and coronal CT; histopathologic and immunohistochemical studies including sections stained with hematoxylin-eosin, periodic acid Schiff, alcian blue, mucicarmine, and Prussian blue for iron; and monoclonal antibodies against cytokeratin-7, epithelial membrane antigen, smooth muscle actin for myoepithelial cells, gross cystic disease fluid protein-15 for apocrine differentiation, and CD-68 and lysozyme for histiocytes. The cyst possessed a multilaminar lining composed of polygonal to low cuboidal cells. A stalk of solid tumor ingrowth from the wall was composed of adenomatous units of eosinophilic cells with apical snouts ("decapitation secretion"). No goblet cells were discovered. Both the cyst's lining cells and the adenoma expressed gross cystic disease fluid protein-15 indicative of apocrine differentiation. The adenoma displayed inner adlumenal cells that were CK-7 and epithelial membrane antigen positive, and outer myoepithelial cells that were smooth muscle actin positive. Simple local excision was curative. This unique lesion is the first example in the ophthalmic and dermatopathologic literatures of a solid adenoma encompassed by an apocrine cyst, which more typically features short or blunt papillae. It must be distinguished from other eyelid and/or anterior orbital cystic lesions including eccrine hydrocystomas; classical and extratarsal dermoid cysts; congenital and acquired conjunctival cysts and dermoids; dacryocystocoeles/mucoceles; canaliculops and lacrimal gland dacryops; and congenital cystic odontogenic choristomas.

  3. Matrix metalloproteinase-1 expression in oral submucous fibrosis: An immunohistochemical study

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    Mishra Gauri

    2010-01-01

    Full Text Available Context: Oral submucous fibrosis (OSF is a form of pathological fibrosis affecting the oral mucosa. There is compelling evidence to implicate the habitual chewing of areca nut with the development of OSF. Because collagens are the major structural components of connective tissues, including oral submucosa, the composition of collagen within each tissue needs to be precisely regulated to maintain tissue integrity. Arecoline stimulates fibroblasts to increase the production of collagen by 150%. Aim: As the role of collagenase is implicated in cleaving the collagen under physical conditions, this study was carried out to evaluate the role of collagenase-1 (matrix metalloproteinase [MMP]-1 in a pathologic condition like OSF. Settings and Design: A total of 40 patients were included in the study, comprising of 30 OSF as Group 1 and 10 normal buccal mucosa tissue as Group 2. Materials and Methods: Both the groups were stained for MMP-1 by the immunohistochemical method using the streptavidin HRP-biotin labeling technique. MMP-1 expression intensity in the epithelium and connective tissue was decreased in Group 1 when compared to Group 2. Statistical Analysis Used: Chi-square test of association was used to determine the difference in the expression of MMP-1 between OSF and normal buccal mucosa and among different histological gradings of OSF. Results: The results were statistically significant. However, there was no statistically significant difference between the expression of MMP-1 among different histological grades of OSF in Group 1.

  4. Histopathological and Immunohistochemical Findings in Congenital Pouch Colon: A Prospective Study.

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    Udawat, Hema; Nunia, Vandana; Mathur, Praveen; Udawat, Harsh Prasad; Gaur, Kusum Lata; Saxena, Amulya K; Mohan, M Krishna

    2017-01-01

    There is a paucity of literature on the histopathological aspects of congenital pouch colon (CPC) and immunohistochemical (IHC) assessment has not been reported. So we planned to study the histopathological and IHC findings within the spectrum of CPC and compare the findings with the normal colon. This is a descriptive prospective study on CPC patients. There were 49 cases of CPC (42 males and 7 females) and 13 controls. Histological examination was done using hematoxylin and eosin and Masson trichrome stain. IHC analysis was done with actin, myosin, and desmin antibodies, and neuron-specific enolase and S100 markers for counting ganglionic cells. Histologically, congestion, edema and hemorrhage were seen in mucosa, submucosa, and serosa. Muscle layers were disrupted and divided into bands. An additional muscle coat inside of the muscularis propria was seen in CPC types 1 and 2. Mature ganglionic cells were reduced and muscle layers showed reduced and patchy positivity for smooth muscle actin, myosin, and desmin compared to a normal colon. Histopathological and IHC findings suggest that CPC has distinct defects in the neuromusculature. © 2017 S. Karger AG, Basel.

  5. Immunohistochemical localization of aquaporin 4 (AQP4 in the porcine gastrointestinal tract

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    Marcin Bartłomiej Arciszewski

    2015-01-01

    Full Text Available The water channel aquaporin-4 (AQP4 is a protein widely expressed on plasma membrane of a variety of epithelial cells. In this study we investigated the expression of AQP4 in the gastrointestinal tract of the pig using immunohistochemical staining. We found no presence of AQP4 in the different regions of the pig stomach. In the porcine small intestine moderate immunoreactivity to AQP4 was detected in enterocytes (along the villi and in the bottom of the crypts, duodenal Brunner’s glands and in enteric ganglia in cells lying in close vicinity to myenteric as well as submucous neurons. In superficial epithelial cells of the colonic mucosa as well as of caecal and colonic glands a very strong immunoreactivity to AQP4 was found. Both in the myenteric and submucous ganglia of the large intestine AQP4-positive cells surrounding enteric neurons were observed. We concluded that AQP4 expression in the porcine gastrointestinal tract showed some species-dependent differences in relation to other species. Based on the presented distribution pattern of AQP4, it is likely that the aquaporin plays a role in mucous (but not acid secretion and intestinal absorptive processes in the pig.

  6. Immunohistochemical characterization of epithelial cells implanted in the flap-stroma interface of the cornea.

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    Chen, Lizhong; Kato, Takuji; Toshida, Hiroshi; Nakamura, Shinji; Murakami, Akira

    2005-01-01

    To investigate the expression of extracellular matrix collagens and their relationship to corneal opacities after implantation of epithelial cells in the flap-stroma corneal interface. A corneal flap was made on rabbit eyes, and epithelial cells, mechanically scraped from tissue surrounding the flap, were implanted beneath the flap. The corneas were harvested 1, 3, 7, and 30 days following surgery. Histological and immunohistochemical examinations were performed. The expression and localization of types I, III, and IV collagens and gelatinase A were determined. Slit-lamp examination showed corneal opacity in the area where the epithelial cells were implanted. Histological study revealed clusters of epithelial cells between the flap and stromal interface. One week and 1 month after the implantation, intense immunoreactivity for collagen type IV was detected at the perimeters of the intrastromal epithelial islands, but not in the interface outside the implanted epithelial cells. Weak positive staining for gelatinase A was detected in the implanted epithelial cells and surrounding keratocytes. The heavy deposition of collagen type IV surrounding the implanted epithelial cells indicated that it might be an essential component of the interface haze observed in patients following laser in situ Keratomileusis. Gelatinase A may also play a role in the regulation of stromal remodeling after epithelial ingrowth.

  7. Immunohistochemical localization of mesenchymal stem cells in ossified human spinal ligaments.

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    Chin, Shunfu; Furukawa, Ken-Ichi; Ono, Atsushi; Asari, Toru; Harada, Yoshifumi; Wada, Kanichiro; Tanaka, Toshihiro; Inaba, Wataru; Mizukami, Hiroki; Motomura, Shigeru; Yagihashi, Soroku; Ishibashi, Yasuyuki

    2013-07-12

    Mesenchymal stem cells (MSCs) have been isolated from various tissues and used for elucidating the pathogenesis of numerous diseases. In our previous in vitro study, we showed the existence of MSCs in human spinal ligaments and hypothesized that these MSCs contributed to the pathogenesis of ossification of spinal ligaments. The purpose of this study was to use immunohistochemical techniques to analyze the localization of MSCs in ossified human spinal ligaments in situ. Ossified (OLF) or non-ossified ligamentum flavum (non-OLF) samples from the thoracic vertebra were obtained from patients who had undergone posterior spinal surgery. Serial sections were prepared from paraffin-embedded samples, and double immunofluorescence staining was performed using antibodies against markers for MSCs (CD73, CD90 and CD105), endothelial cells (CD31), pericytes (α-smooth muscle actin), and chondrocytes (S100). Immunolocalization of MSCs was observed in the perivascular area and collagenous matrix in spinal ligaments. Markers for MSCs and pericytes were co-expressed in the perivascular area. Compared with non-OLF, OLF had a large amount of neovascularization in the fragmented ligament matrix, and a high accumulation of MSCs around blood vessels. The prevalence of MSCs in OLF within collagenous matrix was significantly higher than that in non-OLF. Chondrocytes near the ossification front in OLF also presented expression of MSC markers. MSCs may contribute to the ectopic ossification process of OLF through endochondral ossification. Copyright © 2013 Elsevier Inc. All rights reserved.

  8. Immunohistochemical Expression of p53 in Pleomorphic Adenoma and Carcinoma Ex Pleomorphic Adenoma

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    Bassel Tarakji

    2010-01-01

    Full Text Available Context. Immunohistochemical stains for p53 are used as a diagnostic marker associated with malignancy in several histologic types of salivary gland tumors. This marker may be useful in differentiating pleomorphic adenoma (PA from carcinoma ex pleomorphic adenoma (CPA, as these tumors are often difficult to distinguish on the basis of morphology alone. Objective. to evaluate whatever inactivation of tumor suppressor gene (p53 increases with the tumor progression from normal salivary tissue to PA and eventually CPA. Design. Paraffin blocks of 29 cases of PA, which were surrounded by normal parotid gland, and 27 cases of carcinoma ex pleomorphic adenoma were retrieved and validated. In all cases of carcinoma ex pleomorphic adenoma, a PA “ghost” was identified, and the malignant element was either undifferentiated carcinoma or adenocarcinoma. Results. The results showed negative nuclear expression of P53 in normal parotid gland. Nuclear P53 was expressed strongly in 6/29 (20.7% pleomorphic salivary adenoma and 10/27 (37% carcinoma ex pleomorphic adenoma. Conclusion. Our data suggest that inactivation of p53 may play an important role in the evolution of pleomorphic salivary adenoma and carcinoma ex pleomorphic adenoma.

  9. Chronic moderate alcohol consumption induces iNOS expression in the penis: An immunohistochemical study.

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    Gonca, Süheyla; Yazir, Yusufhan; Göçmez, Semil Selcan; Dalçik, Ekim Nur; Utkan, Tijen; Dalçik, Hakki

    2014-01-01

    To investigate the effect of moderate alcohol consumption on metabolic alterations, inducible nitric oxide synthase (iNOS), immunohistochemical distribution, and morphological damage to penile erectile tissue in rats. Male Wistar albino rats were divided into 2 groups. Group 1 rats (control group, n = 8) received tap water ad libitum, and group 2 rats (n = 8) were fed with 20% ethanol. Increasing levels of alcohol were given to the rats over 12 weeks. Immunohistochemistry was then performed using the avidin-biotin-peroxidase technique on 5-pm thickness tissue sections. Stained sections were examined by imaging microscope. Alcohol consumption resulted in a significant increase in iNOS immunoreactivity in the penile erectile tissue. Increased iNOS expression was determined in the tunica albuginea, cavernosal smooth muscle cells, trabeculae of connective tissue, arterioles, and the urethral epithelium. Moreover, chronic alcohol consumption resulted in decreasing serum testosterone and high density lipoprotein (HDL) levels with increasing cholesterol and triglyceride levels. Chronic moderate alcohol consumption can affect penile erectile tissue by increasing iNOS immunoreactivity and induce histopathological damage such as penile fibrosis. These abnormalities are also related to the defense mechanism against morphological damage.

  10. Detection of Cancer Stem Cells in Colorectal Cancer: Histopathological and Immunohistochemical Study

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    Nour El Hoda S. Ismaiel

    2016-12-01

    Full Text Available BACKGROUND: Growing evidence supports the notion that the onset of tumorigenesis could occur through cancer stem cells (CSCs. These tumour cells show low proliferative rates, high self-renewal capacity, propensity to differentiate into active proliferating tumour cells & resistance to chemoradiotherapy thus, possibly causing local recurrences & metastasis formation. CD44 has been used as a marker to isolate CSCs from colorectal carcinoma (CRC. AIM: To investigate the immunohistochemical expression of cancer stem cells marker (CD44 in CRC and correlate its expression with the clinicopathological aspects, TNM staging and modified Dukes’ classification. MATERIALS AND METHODS: Tumour biopsies from colectomy specimens of 60 patients with CRC were stained with hematoxylin-eosin for histological evaluation then immunostained with monoclonal antibodies against CD44 which was detected in term of negative or positive expression. RESULTS: CD44 was demonstrated in 58.3% (35/60 of cases and showed statistically significant correlation with tumour site and histological type (p-value 0.05. Chi-square /Fisher exact test proportion independence and the p-value are set significant at 0.05 level. CONCLUSION: the CD44 rate of expression is higher in the colon than rectum and in adenocarcinoma than mucinous and undifferentiated carcinoma. CD44 showed statistically insignificant relation with T, N, M, grade, TNM stage grouping and modified Dukes’ classification.

  11. Immunohistochemical Studies on Galectin Expression in Colectomised Patients with Ulcerative Colitis

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    Mattias Block

    2016-01-01

    Full Text Available 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.

  12. Immunohistochemical distribution of Plexin A4 in the adult rat central nervous system

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    Claire-Anne Gutekunst

    2010-07-01

    Full Text Available PlexinA4 is the latest member to be identified of the plexin A subfamily, critical transducers of class 3 semaphorin signaling as co-receptors to neuropilins 1 and 2. Despite functional information regarding the role of PlexinA4 in development and guidance of specific neuronal pathways, little is known about its distribution in the adult central nervous system (CNS. Here we report an in depth immunohistochemical analysis of PlexinA4 expression in the adult rat CNS. PlexinA4 staining was present in neurons and fibers throughout the brain and spinal cord, including neocortex, hippocampus, lateral hypothalamus, red nucleus, facial nucleus and the mesencephalic trigeminal nucleus. PlexinA4 antibodies labeled fibers in the lateral septum, nucleus accumbens, several thalamic nuclei, substantia nigra pars reticulata, zona incerta, pontine reticular region, as well as in several cranial nerve nuclei. This constitutes the first detailed description of the topographic distribution of PlexinA4 in the adult CNS and will set the basis for future studies on the functional implications of PlexinA4 in adult brain physiology.

  13. Delineation of Chondroid Lipoma: An Immunohistochemical and Molecular Biological Analysis

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    Ronald S. A. de Vreeze

    2011-01-01

    Full Text Available Aims. Chondroid lipoma (CL is a benign tumor that mimics a variety of soft tissue tumors and is characterized by translocation t(11;16. Here, we analyze CL and its histological mimics. Methods. CL (n=4 was compared to a variety of histological mimics (n=83 for morphological aspects and immunohistochemical features including cyclinD1(CCND1. Using FISH analysis, CCND1 and FUS were investigated as potential translocation partners. Results. All CLs were strongly positive for CCND1. One of 4 myoepitheliomas, CCND1, was positive. In well-differentiated lipomatous tumors and in chondrosarcomas, CCND1 was frequently expressed, but all myxoid liposarcomas were negative. FISH analysis did not give support for direct involvement of CCND1 and FUS as translocation partners. Conclusions. Chondroid lipoma is extremely rare and has several and more prevalent histological mimics. The differential diagnosis of chondroid lipomas can be unraveled using immunohistochemical and molecular support.

  14. Ileal ganglioneuromatosis in a piglet: histopathological and immunohistochemical studies.

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    Quiroga, M A; Lozada, M I; Madariaga, G; Cappucio, J A; Machuca, M A; Barrales, H; Perez, E M; Perfumo, C J

    2014-11-01

    Ganglioneuromatosis (GNM) is a rare condition characterized by the benign proliferation of ganglion cells, nerve fibres and supporting cells of the enteric nervous system. Necropsy examination of a female piglet weighing 4 kg revealed a well-demarcated 20 cm segment of terminal ileum with thickening of the wall. Microscopically, the lamina propria was infiltrated by enteric glial cells and large ganglion cells. Within the submucosal and muscular layers, aggregates of neurons were interlaced by Schwann cells and enteric glial cells arranged in concentric rings. Immunohistochemically, the neurons were weakly labelled for S-100 and neuron-specific enolase, Schwann cells expressed S-100 and vimentin and enteric glial cells expressed glial fibrillary acidic protein and S-100. Pathological and immunohistochemical findings supported the diagnosis of ileal GNM. Copyright © 2014 Elsevier Ltd. All rights reserved.

  15. Investigation of telomerase activity and apoptosis on invasive ductal carcinoma of the breast using immunohistochemical and Western blot methods.

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    Simsek, B C; Turk, B A; Ozen, F; Tuzcu, M; Kanter, M

    2015-08-01

    Invasive ductal carcinoma (IDC) comprises the largest group of breast cancers. This study aimed to investigate telomerase activity and apoptosis using immunohistochemical and Western blot methods. In total, 75 cases that had been diagnosed as IDC and 20 cases that had undergone a freezing procedure were included. The histological sections were stained with Bax, Bcl-2, hTERT and BNIP3. The ages of the patients, as well as their hormonal status and tumour sizes and grades were evaluated, as well as the staining characteristics of the antibodies in question. A decrease in Bcl-2 positivity and an increase in Bax positivity were found immunohistochemically with increasing tumour grades. The data obtained by western blot method showed that Bcl-2 was highest in grade 1 tumours although these results were not statistically significant. The relationship between estrogen and progesterone receptor positivity and Bcl-2 was statistically significant, suggesting there is hormonal control through apoptosis. BNIP3 was found to be decreased with increasing tumour grades. Similarly, BNIP3 was found to be having the lowest value in grade 3 tumours by western blot method. Furthermore, hTERT was found to be increased with increasing tumour grades. In the western blot method, hTERT increased nearly four-fold compared to the control. In addition, hTERT, which was seen in very high levels in tumours, may be a helpful cancer marker. Both hTERT and BNIP3 are important markers that can provide information about prognosis. Big improvements can be achieved in tumour progression control with new treatment modalities that stop telomerase activity and hypoxic cell death.

  16. Immunohistochemical study of polycystin-1 in dentigerous cysts

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    Jaishlal Mariachelliah Nadar Singarayan

    2014-01-01

    Discussion and Conclusion: All PC1 positive epithelial cells′ cytoplasm stained diffusely. Abnormal cytoplasmic expression of PC1 in all positive epithelial lining indicates that the PC1 probably is associated with cystic transformation.

  17. Immunohistochemical Evaluation of Mononuclear Infiltrates in Canine Lupoid Onychodystrophy

    National Research Council Canada - National Science Library

    Mueller, R. S; West, K; Bettenay, S. V

    2004-01-01

    Claw biopsy samples of 11 dogs with lupoid onychodystrophy were evaluated. They were stained with hematoxylin and eosin and with antibodies against CD 3 as a T-cell marker, BLA 36 and HM 57 (CD 79α...

  18. Immunohistochemical findings of the granulomatous reaction associated with tuberculosis

    Directory of Open Access Journals (Sweden)

    Shirin Karimi

    2016-01-01

    Conclusion: Detection of TB in tissue slides is still based on the histological pattern of the granuloma, which has several differential diagnoses with different treatments. Presence of mycobacterial antigens and tissue morphology can be evaluated using the IHC technique. Considering the criteria of positive IHC staining of TB granulomatous reactions, this stain not only highlights the presence of mycobacterial antigens for tissue diagnosis, but also could morphologically localize their distribution in different cells. Pathologists must be familiar with adequate staining pattern, elimination of background staining, and type of selected antibody. This method is especially important for application in countries with high prevalence of TB as a technique with early diagnostic value in tissue specimens. Early diagnosis using this technique can reduce related morbidity and mortality and decrease the rate of complications due to misdiagnosis and mistreatment of TB.

  19. A simplified immunohistochemical classification of skeletal muscle fibres in mouse

    OpenAIRE

    Kammoun, M.; Cassar-malek, I.; Meunier, B; Picard, B.

    2014-01-01

    The classification of muscle fibres is of particular interest for the study of the skeletal muscle properties in a wide range of scientific fields, especially animal phenotyping. It is therefore important to define a reliable method for classifying fibre types. The aim of this study was to establish a simplified method for the immunohistochemical classification of fibres in mouse. To carry it out, we first tested a combination of several anti myosin heavy chain (MyHC) antibodies in order to c...

  20. A simplified immunohistochemical classification of skeletal muscle fibres in mouse

    Directory of Open Access Journals (Sweden)

    M. Kammoun

    2014-06-01

    Full Text Available The classification of muscle fibres is of particular interest for the study of the skeletal muscle properties in a wide range of scientific fields, especially animal phenotyping. It is therefore important to define a reliable method for classifying fibre types. The aim of this study was to establish a simplified method for the immunohistochemical classification of fibres in mouse. To carry it out, we first tested a combination of several anti myosin heavy chain (MyHC antibodies in order to choose a minimum number of antibodies to implement a semi-automatic classification. Then, we compared the classification of fibres to the MyHC electrophoretic pattern on the same samples. Only two anti MyHC antibodies on serial sections with the fluorescent labeling of the Laminin were necessary to classify properly fibre types in Tibialis Anterior and Soleus mouse muscles in normal physiological conditions. This classification was virtually identical to the classification realized by the electrophoretic separation of MyHC. This immunohistochemical classification can be applied to the total area of Tibialis Anterior and Soleus mouse muscles. Thus, we provide here a useful, simple and time-efficient method for immunohistochemical classification of fibres, applicable for research in mouse

  1. Reduced immunohistochemical expression of adhesion molecules in vitiligo skin biopsies.

    Science.gov (United States)

    Reichert Faria, Adriane; Jung, Juliana Elizabeth; Silva de Castro, Caio César; de Noronha, Lucia

    2017-03-01

    Because defects in adhesion impairment seem to be involved in the etiopathogenesis of vitiligo, this study aimed to compare the immunohistochemical expression of several adhesion molecules in the epidermis of vitiligo and non lesional vitiligo skin. Sixty-six specimens of lesional and non lesional skin from 33 volunteers with vitiligo were evaluated by immunohistochemistry using anti-beta-catenin, anti-E-cadherin, anti-laminin, anti-beta1 integrin, anti-collagen IV, anti-ICAM-1 and anti-VCAM-1 antibodies. Biopsies of vitiligo skin demonstrated a significant reduction in the expression of laminin and integrin. The average value of the immunohistochemically positive reaction area of the vitiligo specimens was 3053.2μm2, compared with the observed value of 3431.8μm2 in non vitiligo skin (p=0.003) for laminin. The immuno-positive area was 7174.6μm2 (vitiligo) and 8966.7μm2 (non lesional skin) for integrin (p=0.042). A reduction in ICAM-1 and VCAM-1 expression in the basal layer of the epidermis in vitiligo samples was also observed (p=0.001 and pvitiligo and non lesional skin. Our results suggest that an impairment in adhesion exists in vitiligo skin, which is supported by the diminished immunohistochemical expression of laminin, beta1 integrin, ICAM-1 and VCAM-1. Copyright © 2017 Elsevier GmbH. All rights reserved.

  2. A simplified immunohistochemical classification of skeletal muscle fibres in mouse.

    Science.gov (United States)

    Kammoun, M; Cassar-Malek, I; Meunier, B; Picard, B

    2014-06-24

    The classification of muscle fibres is of particular interest for the study of the skeletal muscle properties in a wide range of scientific fields, especially animal phenotyping. It is therefore important to define a reliable method for classifying fibre types. The aim of this study was to establish a simplified method for the immunohistochemical classification of fibres in mouse. To carry it out, we first tested a combination of several anti myosin heavy chain (MyHC) antibodies in order to choose a minimum number of antibodies to implement a semi-automatic classification. Then, we compared the classification of fibres to the MyHC electrophoretic pattern on the same samples. Only two anti MyHC antibodies on serial sections with the fluorescent labeling of the Laminin were necessary to classify properly fibre types in Tibialis Anterior and Soleus mouse muscles in normal physiological conditions. This classification was virtually identical to the classification realized by the electrophoretic separation of MyHC. This immunohistochemical classification can be applied to the total area of Tibialis Anterior and Soleus mouse muscles. Thus, we provide here a useful, simple and time-efficient method for immunohistochemical classification of fibres, applicable for research in mouse.

  3. Comparison of tetrachromic VOF stain to other histochemical staining techniques for characterizing stromal soft and hard tissue components.

    Science.gov (United States)

    Belaldavar, C; Hallikerimath, S; Angadi, P V; Kale, A D

    2014-11-01

    The components of hard tissues including dentin, enamel, cementum, bone and other calcified deposits, and mature and immature collagen pose problems for identification in routine hematoxylin and eosin (H & E) stained sections. Use of combinations of stains can demonstrate the components of hard tissues and soft tissues distinctly. We assessed the efficacy of the Verde Luz-orange G-acid fuchsin (VOF) stain for differentiating hard and soft connective tissues and compared results with other histochemical staining techniques. Eighty tissue sections comprising developing tooth (30), ossifying fibroma (30) and miscellaneous pathologies (20) expected to contain varying types of calcified tissues were stained with H & E, VOF, and Masson's trichrome (MT). In developing tooth, VOF demonstrated better differentiation of hard tissues, while it was comparable to MT for ossifying fibroma and miscellaneous pathologies. The intensity of staining was greater with VOF than with the other stains studied. VOF stains hard tissue components distinctly and gives good contrast with the surrounding connective tissue. VOF is comparable to MT, but has added advantages including single step staining, rapid and easy procedures, and it distinguishes the maturity of the tissues.

  4. News from the Biological Stain Commission no. 12

    DEFF Research Database (Denmark)

    Lyon, H O

    2012-01-01

    In this 12(th) issue of News from the Biological Stain Commission (BSC) under the heading of Regulatory affairs, the Biological Stain Commission's International Affairs Committee presents information from the meetings of ISO/TC 212/WG 1 Quality and competence in the medical laboratory and ISO...

  5. Standardization in biological staining. The influence of dye manufacturing

    DEFF Research Database (Denmark)

    Lyon, H

    2000-01-01

    for biological staining, standardization of all procedures and reagents is mandatory. In this paper, I focus particularly on dyes and consider the possibilities for obtaining standardized dyes. In general practice, most biological staining takes place with available commercial dyes. These dyes may or may...

  6. Diagnostic utility of the genital Gram stain in ED patients.

    Science.gov (United States)

    Stefanski, Peter; Hafner, John W; Riley, Shanda L; Sunga, Kharmene L Y; Schaefer, Timothy J

    2010-01-01

    The study aimed to determine the diagnostic usefulness of the genital Gram stain in an emergency department (ED) population. A linked-query of an urban, tertiary-care, university- affiliated hospital laboratory database was conducted for all completed Chlamydia trachomatis and Neisseria gonorrhoeae DNA probes, Trichomonas vaginalis wet preps, and genital Gram stains performed on ED patient visits between January and December 2004. Positive criteria for a Gram stain included greater than 10 white blood cells per high-power field, gram-negative intracellular/extracellular diplococci (suggesting N gonorrhoeae), clue cells (suggesting T vaginalis), or direct visualization of T vaginalis organisms. DNA probes were used as the gold standard definition for N gonorrhoeae and C trachomatis infection. Of 1511 initially eligible ED visits, 941 were analyzed (genital Gram stain and DNA probe results both present), with a prevalence of either C trachomatis or N gonorrhoeae of 11.4%. A positive genital Gram stain was 75.7% sensitive and 43.3% specific in diagnosing either C trachomatis and/or N gonorrhoeae infection, and 80.4% sensitive and 32.2% specific when the positive cutoff was lowered to more than 5 white blood cells/high-power field. No Gram stains were positive for T vaginalis (with 47 positive wet mounts), and clue cells were noted on 117 Gram stains (11.6%). Gram stains in isolation lack sufficient diagnostic ability to detect either C trachomatis or N gonorrhoeae infection in the ED.

  7. The use of special stains in liver biopsy interpretation: Implications ...

    African Journals Online (AJOL)

    Materials and Methods: The formalin fixed paraffin embedded blocks of liver biopsies reported in two histopathology laboratories between 2008 and 2013 were retrieved. These were stained with H and E and the following standard special stains for liver tissue histology – Perl's Prussian blue, reticulin, Sirius red, Shikata ...

  8. Lawsonia inermis And Hibiscus sabdariffa : Posible Histological Stains

    African Journals Online (AJOL)

    The ability of various concentrations of aqueous extracts of Lawsonia inermis and Hibiscus sabdariffa to stain histological tissues was demonstrated. The results with sections of tongue and kidney of the laboratory rat, cut at 6microns thickness showed that only the cellular cytoplasm was stained. However, combinations of ...

  9. Are port wines stains a feature of tuberous sclerosis?

    Science.gov (United States)

    Ben-Amitai, D; Halachmi, S; Lapidoth, M

    2011-07-01

    Tuberous sclerosis complex is a multisystem inherited disorder characterized by the development of tumour-like growths in brain, skin and other organs. Although cutaneous vascular anomalies are not considered a common manifestation, we have encountered co-occurrence of port wine stains and tuberous sclerosis. To assess the prevalence of port wine stain in patients with previously diagnosed tuberous sclerosis. All cases diagnosed with tuberous sclerosis at two tertiary care centres from 2000 to 2009 were reviewed. Cases with clinically documented port wine stains were included for evaluation. Of 24 patients diagnosed with tuberous sclerosis, three (12.5%) had clinically evident port wine stains. The prevalence of port wine stains in this series of tuberous sclerosis patients was significantly higher than the 0.3% prevalence of port wine stain in the general population. Port wine stain rate in this population was significantly greater than the expected rate. Further studies are needed to assess the frequency of port wine stains in tuberous sclerosis and to clarify whether the finding should be added to the list of cutaneous features of tuberous sclerosis. © 2010 The Authors. Journal of the European Academy of Dermatology and Venereology © 2010 European Academy of Dermatology and Venereology.

  10. THIONIN STAINING OF PARAFFIN AND PLASTIC EMBEDDED SECTIONS OF CARTILAGE

    NARCIS (Netherlands)

    BULSTRA, SK; DRUKKER, J; KUIJER, R; BUURMAN, WA; VANDERLINDEN, AJ

    The usefulness of thionin for staining cartilage sections embedded in glycol methacrylate (GMA) and the effect of decalcification on cartilage sections embedded in paraffin and GMA were assessed. Short decalcification periods using 5% formic acid or 10% EDTA did not influence the staining properties

  11. AUTHENTIC MATERIALS IN EXTENSIVE READING CLASS AT STAIN PONOROGO

    Directory of Open Access Journals (Sweden)

    Dhinuk Puspita Kirana

    2013-12-01

    Full Text Available It is widely believed that English Foreign Language (EFL learners need to develop their language proficiency by getting so much input. Moreover, students need to be familiarized with the real English us­age where real forms of communication and cultural knowledge are crucially exposed. Teaching through authentic materials will make the learners feel that they are learning a real language which is used by the real native speakers for real communication. incorporating au­thentic materials helps students acquire an effective communicative competence in the language focus. The research intended to describe the implementation of authentic materials in extensive reading class, the problems arise and the students’ responses toward the authen­tic materials in extensive reading class. The design of the research was Descriptive Qualitative method and the research subject was the lecturer of Extensive Reading class and 33 students in B class of the fourth semester of STAIN Ponorogo who took Extensive Read­ing subject. The instruments used were in the form of observation sheet, interview guideline and questionnaire. The implementation of authentic materials in extensive reading class covered some procedures into three main phases namely (1 Pre­ Activity, (2 Main­ Activity and (3 Post­Activity. The activities in main activity are as follows: (a Pre­ Activity; (b Whilst ­Activity; and (3 The language focus stage. There were problems arose during the implementation in terms of complicated planning, more time allocation and some disinterested students. Finally, the students showed significantly positive attitude toward the implementation of authentic materials in extensive reading class.

  12. Stereomicroscopic examination of stained rectal biopsies in ulcerative colitis and Crohn's disease

    DEFF Research Database (Denmark)

    Poulsen, Steen Seier

    1980-01-01

    Rectal biopsy samples from 22 healthy control subjects, 54 patients with ulcerative colitis, and 34 with Crohn's disease with involvement of the colon or rectum were investigated in a stereomicroscopic study. Samples were stained as whole mounts with Alcian Green before the stereomicroscopic...... the stereomicroscopic findings and the clinical disease activity, the sigmoidoscopic findings, and the histologic activity. Apart from the stereomicroscopic observation of small superficial erosions in one fourth of the biopsies, no changes of diagnostic value were observed in Crohn's disease....

  13. Mapping stain distribution in pathology slides using whole slide imaging

    Directory of Open Access Journals (Sweden)

    Fang-Cheng Yeh

    2014-01-01

    Full Text Available Background: Whole slide imaging (WSI offers a novel approach to digitize and review pathology slides, but the voluminous data generated by this technology demand new computational methods for image analysis. Materials and Methods: In this study, we report a method that recognizes stains in WSI data and uses kernel density estimator to calculate the stain density across the digitized pathology slides. The validation study was conducted using a rat model of acute cardiac allograft rejection and another rat model of heart ischemia/reperfusion injury. Immunohistochemistry (IHC was conducted to label ED1 + macrophages in the tissue sections and the stained slides were digitized by a whole slide scanner. The whole slide images were tessellated to enable parallel processing. Pixel-wise stain classification was conducted to classify the IHC stains from those of the background and the density distribution of the identified IHC stains was then calculated by the kernel density estimator. Results: The regression analysis showed a correlation coefficient of 0.8961 between the number of IHC stains counted by our stain recognition algorithm and that by the manual counting, suggesting that our stain recognition algorithm was in good agreement with the manual counting. The density distribution of the IHC stains showed a consistent pattern with those of the cellular magnetic resonance (MR images that detected macrophages labeled by ultrasmall superparamagnetic iron-oxide or micron-sized iron-oxide particles. Conclusions: Our method provides a new imaging modality to facilitate clinical diagnosis. It also provides a way to validate/correlate cellular MRI data used for tracking immune-cell infiltration in cardiac transplant rejection and cardiac ischemic injury.

  14. Effect of formalin fixation on the immunohistochemical detection of PRRS virus antigen in experimentally and naturally infected pigs.

    Science.gov (United States)

    Van Alstine, W G; Popielarczyk, M; Albregts, S R

    2002-11-01

    The purpose of this study was to determine the effect of formalin fixation on the immunohistochemical detection of porcine reproductive and respiratory syndrome (PRRS) viral antigen in lungs of experimentally and naturally infected pigs. In separate trials, five 24-day-old pigs and six 10-day-old pigs were housed as separate groups in isolation and inoculated intranasally with 10(5.5) TCID50 of an isolate of PRRS virus (PRRSV; P129). The older and younger pigs were euthanatized at 7 and 10 days post inoculation (dpi), respectively. At necropsy, all pigs had gross and microscopic lung lesions typical of PRRS, and PRRSV was isolated from all pigs. To insure uniform fixation, lungs from each pig were cut into 1-cm-thick slices and immersed into 10% neutral-buffered formalin. After fixation in formalin for 8 hours or 1, 2, 3, 5, 6, 8, 10, and 15 days, 3 lung sections from some or all pigs were processed for histological examination using routine methods. Immunohistochemical staining for PRRSV antigen was positive at the following times (days unless otherwise stated) after fixation (percentage of pigs staining positive for PRRSV in parentheses): 8 hours (100); 1 (100); 2 (100); 3 (80); 5 (33); and 6, 8, 10, and 15 (0-all negative). To further evaluate the effects of formalin fixation on PRRSV immunodetection, 31 field cases of PRRS were selected for immunohistochemistry (IHC). Over a 3-month period, submitted cases were selected from the Purdue University Animal Disease Diagnostic Laboratory, W. Lafayette, Indiana, for IHC if 1) the clinical history included respiratory disease, 2) PRRSV was isolated from lung and/or serum from the submitted pigs or tissues, 3) at least 1 section of lung fixed in 10% neutral-buffered formalin was submitted for IHC, and 4) the duration of fixation could be accurately determined from the case history. Of the 31 PRRSV-infected pig cases meeting the selection criteria, 23 were fixed in formalin for 4 days or less. Twenty-one of these 23 (91

  15. Enhanced Port Wine Stain Lightening Achieved with Combined Treatment of Selective Photothermolysis and Imiquimod

    Science.gov (United States)

    Tremaine, Anne Marie; Armstrong, Jennifer; Huang, Yu-Chih; Elkeeb, Leila; Ortiz, Arisa; Harris, Ronald; Choi, Bernard; Kelly, Kristen M.

    2012-01-01

    Background Pulsed dye laser is the gold standard for treatment of port wine stain birthmarks but multiple treatments are required and complete resolution is often not achieved. Post-treatment vessel recurrence is thought to be a factor that limits efficacy of pulsed dye laser treatment of port wine stains. Imiquimod 5% cream is an immunomodulator with anti-angiogenic effects. Objective To determine if application of imiquimod 5% cream after pulsed dye laser improves treatment outcome. Methods Healthy patients with port wine stains (n = 24) were treated with pulsed dye laser and then randomized to apply post-treatment placebo or imiquimod 5% cream for 8 weeks. Chromameter measurements (CIE L*a*b* colorspace) for 57 port wine stain sites (multiple sites per subject) were taken at baseline and compared with measurements taken 8 weeks post-treatment. The change in a* and ΔE were measured to quantify treatment outcome. Results Two subjects developed minor skin irritation. Other adverse effects weren't noted. Average Δa* was 0.43 for pulsed dye laser + placebo sites (n = 25) and 1.27 for pulsed dye laser + imiquimod sites (n = 32) (p value = 0.0294) indicating a greater reduction in erythema with imiquimod. Average ΔE was 2.59 for pulsed dye laser + placebo and 4.08 for pulsed dye laser + imiquimod (p value = 0.0363), again indicating a greater color improvement with imiquimod. Limitations Effects were evaluated after a single treatment and duration of effect is unknown. Conclusion Combined selective photothermolysis and anti-angiogenic therapy may enhance port wine stain treatment efficacy. PMID:22244840

  16. Novel Process for Laser Stain Removal from Archaeological Oil Paintings

    Science.gov (United States)

    El-Nadi, Lotfia; El-Feky, Osama; Abdellatif, Galila; Darwish, Sawsan

    2013-03-01

    Some samples of oil paintings (5 × 5 cm) were prepared on wooden panel with four types of fungi commonly encountered on oil paintings were selected for this study. Each of the fungi is associated with different colored stains. Fungus Alternaria tenuis is associated by a dense black stain, Chetomium globosum by a brownish gray stain, Aspergillus flavus by a yellowish stain, and Fusaruim oxysporum by a pinkish stain. Fungi growing on oil paintings affect the surface characteristics by forming a variety of colored patches typically composed of many complex chemical substances that are produced during metabolic processes. These colored stains may be encrusted in spores, present in mycelium or secreted to a substance such as oil paintings surfaces. While the fungal stains can sometimes be extracted with appropriate solvents, there are some stains that resist solvent extraction entirely. Developing new solvent system that might attack the paint structure, and is time consuming and requires a great deal of trial and error. Mechanical stain removal is also problematic in that it often produces abrasion of the surface, markedly deteriorating the artwork, and is extra ordinarily fine and tedious. For these reasons, we decided to examine an alternative physical technique as a new approach to deal with stain removal. Since the stains are due to the existence of fungi, we thought it a good idea to remove them by singlet oxygen. We applied the photo dynamic process through which the fungi stains were covered with organic dye derivatives in solution under controlled illumination in the lab. The samples were then irradiated by low power Laser light from a He-Ne laser, the dye will be photodecomposed and produce singlet oxygen. We report in this work the results obtained as a function of: - The concentration and types of the organic dye in solution, - The presence of certain amounts of liquids added to the solution, - The scanning speed of the laser beam on the sample surface

  17. Combined bromodeoxyuridine immunohistochemistry and Masson trichrome staining: facilitated detection of cell proliferation in viable vs. infarcted myocardium.

    Science.gov (United States)

    Lazarous, D F; Shou, M; Unger, E F

    1992-09-01

    Cells in the S-phase of the cell cycle can be identified in tissue sections by immunohistochemical localization of the thymidine analogue bromodeoxyuridine (BrdU). Generally, a single counterstain is used to visualize the underlying tissue; however, interpretation of morphologic detail is often difficult. We have utilized BrdU to localize proliferating cells in myocardium exposed to angiogenic mitogens. To facilitate identification of labelled nuclei in the context of infarcted vs. viable myocardium, BrdU immunohistochemistry was followed by a modified Masson trichrome stain. The time of exposure to the counterstains and the wash protocol were re-revised, permitting clear identification of the labelled brown nuclei against a background of red viable myocardium vs. blue infarct. The combined technique also provides color contrast suitable for computer-based image analysis.

  18. A novel washing algorithm for underarm stain removal

    Science.gov (United States)

    Acikgoz Tufan, H.; Gocek, I.; Sahin, U. K.; Erdem, I.

    2017-10-01

    After contacting with human sweat which comprise around 27% sebum, anti-perspirants comprising aluminium chloride or its compounds form a jel-like structure whose solubility in water is very poor. In daily use, this jel-like structure closes sweat pores and hinders wetting of skin by sweat. However, when in contact with garments, they form yellowish stains at the underarm of the garments. These stains are very hard to remove with regular machine washing. In this study, first of all, we focused on understanding and simulating such stain formation on the garments. Two alternative procedures are offered to form jel-like structures. On both procedures, commercially available spray or deo-stick type anti-perspirants, standard acidic and basic sweat solutions and artificial sebum are used to form jel-like structures, and they are applied on fabric in order to get hard stains. Secondly, after simulation of the stain on the fabric, we put our efforts on developing a washing algorithm specifically designed for removal of underarm stains. Eight alternative washing algorithms are offered with varying washing temperature, amounts of detergent, and pre-stain removal procedures. Better algorithm is selected by comparison of Tristimulus Y values after washing.

  19. Western Blot of Stained Proteins from Dried Polyacrylamide Gels

    Science.gov (United States)

    Gruber, Claudia; Stan-Lotter, Helga

    1996-01-01

    Western blotting of proteins is customarily performed following their separation on polyacrylamide gels, either prior to staining (1) or, as recently reported, following staining (2). We describe here Western blotting with stained gels, which had been dried and some of which had been stored for years. This procedure permits immunological analysis of proteins, to which antisera may have become available only later, or where the application of newly developed sensitive detection methods is desired. Once rehydration of the gels is achieved, proteins can be-transferred to blotting membranes by any appropriate protocol. Proteins stained with Coomassie Blue have to be detected with a non-chromogenic method, such as the film-based enhanced chemiluminescence (ECL)2) procedure (3). Silver stained proteins, which transfer in the colorless form, may be visualized by any detection method, although, because of the usually very low amounts of proteins, detection by ECL is preferable. Blotting of stained proteins from rehydrated gels is as rapid and as quantitative as from freshly prepared gels, in contrast to blotting from wet stained gels, which requires extensive washing and results in low transfer efficiency (2). Together with a photographic record of the gel pattern, unambiguous identification of immunoreactive proteins from complex mixtures is possible. Some further applications of this work are discussed.

  20. Microscopic analysis of MTT stained boar sperm cells

    Directory of Open Access Journals (Sweden)

    B.M. van den Berg

    2015-06-01

    Full Text Available The ability of sperm cells to develop colored formazan by reduction of MTT was used earlier to develop a spectrophotometric assay to determine the viability of sperm cells for several mammalian species. It was the objective of the present study to visualize microscopically the location of the formazan in boar sperm cells. The MTT staining process of boar sperm cells can be divided into a series of morphological events. Incubation of the sperm cells in the presence of MTT resulted after a few min in a diffuse staining of the midpiece of the sperm cells. Upon further incubation the staining of the midpiece became more intense, and gradually the formation of packed formazan granules became more visible. At the same time, a small formazan stained granule appeared medially on the sperm head, which increased in size during further incubation. After incubation for about 1 h the midpiece granules were intensely stained and more clearly distinct as granules, while aggregation of sperm cells occurred. Around 90% of the sperm cells showed these staining events. At the end of the staining the formazan granules have disappeared from both the sperm cells and medium, whereas formazan crystals appeared as thin crystal threads, that became heavily aggregated in the incubation medium. It was concluded that formazan is taken up by lipid droplets in the cytoplasm. Further, the use of the MTT assay to test for sperm viability should be regarded as a qualitative assay, whereas its practical use at artificial insemination (AI Stations is limited.

  1. Effect of Melamine Sponge on Tooth Stain Removal.

    Science.gov (United States)

    Otsuka, Takero; Kawata, Toshitsugu

    2015-01-01

    To investigate the stain removal ability of melamine sponge before aesthetic tooth whitening in extracted teeth. Melamine sponge of thickness 40 mm was compressed and the destruction of the partition wall structure during the compression process was examined under a stereoscopic microscope. An extracted human tooth was cleaned by normal polishing or with melamine sponge for 90 s. To evaluate the stain level, the tooth surfaces were photographed under a stereoscopic microscope at 0, 30, 60 and 90 s. The residual stained region was traced in a high-magnification photograph, and the stain intensity was presented as a change, relative to the intensity before the experiment (0 s). Mechanical cleaning by toothbrushing produced polishing scratches on the tooth surface, whereas use of the melamine sponge resulted in only minimal scratches. As the compression level increased, the stain-removing effect tended to become stronger. Melamine sponge can remove stains from the tooth surface more effectively and less invasively compared to a conventional toothbrush. As no new scratches are made on the tooth surface when using a melamine sponge brush, the risk of re-staining is reduced. Cleaning using a melamine sponge brush can be easily and effectively performed at home and in a dental office.

  2. Extrinsic tooth stain removal efficacy of a sodium bicarbonate dual-phase dentifrice containing calcium and phosphate in a six-week clinical trial.

    Science.gov (United States)

    Putt, Mark S; Milleman, Jeffery L; Ghassemi, Annahita

    2004-01-01

    The objective of this clinical investigation was to evaluate the effectiveness in removing existing extrinsic tooth stains of a sodium bicarbonate (baking soda), dual-phase dentifrice containing calcium and phosphate (Test Dentifrice), when compared to a commercial hydrated silica dentifrice (Control Dentifrice), during a six-week period of normal, unsupervised use. This investigation was a double-blind comparison of two equivalent, parallel groups of subjects assigned to use different dentifrices for six weeks. A total of 108 adult male and female subjects qualified for the trial based on the presence of existing extrinsic tooth stains and other inclusion/exclusion criteria. The two groups were balanced for gender, tobacco use, and extrinsic tooth stain scores, and randomly assigned the Test or Control dentifrices. All subjects were instructed to brush their teeth twice daily using only the dentifrice and toothbrush provided, and to refrain from using any other oral hygiene products for the duration of the study. Although product usage was unsupervised after the baseline visit, subjects maintained a treatment diary, and product consumption was monitored to estimate compliance. Oral soft and hard tissue and extrinsic stain assessments (Modified Lobene Stain Index) for each subject were conducted at baseline and after two, four, and six weeks of product use. A total of 107 subjects complied with the protocol and completed the six-week study. Compared to baseline, at the two-, four-, and six-week examinations the Test Dentifrice group had statistically significant reductions in extrinsic tooth stain. In contrast, the Control Dentifrice group did not attain significant reductions from baseline at any of the exams. At both the four-week and six-week examinations, the Test Dentifrice group had significantly lower levels of extrinsic tooth stain than the Control Dentifrice group. A sodium bicarbonate, dual-phase dentifrice containing calcium and phosphate demonstrated

  3. Toothpastes containing abrasive and chemical whitening agents: efficacy in reducing extrinsic dental staining.

    Science.gov (United States)

    Soares, Cristina Neves Girao Salgado; Amaral, Flavia Lucisano Botelho do; Mesquita, Marcelo Ferraz; Franca, Fabiana Mantovani Gomes; Basting, Roberta Tarkany; Turssi, Cecilia Pedroso

    2015-01-01

    This in vitro study evaluated the efficacy of toothpastes containing abrasive and chemical whitening agents in reducing the extrinsic discoloration of dental enamel. Sixty slabs of dentin from human teeth were sealed so that only the enamel surface was exposed. The enamel surfaces were photographed for initial color assessment. Staining was performed by immersing the dental slabs in 0.2% chlorhexidine solution for 2 minutes and then in black tea for 60 minutes. This process was repeated 15 times. Photographs were taken at the end of the staining process, and the slabs were divided into 5 groups (n = 12), 3 to be brushed with toothpastes containing chemical whitening agents (2 containing phosphate salts and 1 containing phosphate salts plus hydrogen peroxide) and 2 to represent control groups (ordinary/nonwhitening toothpaste and distilled water). The dental slabs were subjected to mechanical toothbrushing with toothpaste slurry or distilled water, according to each group's specifications. After brushing, more photographs were taken for color analysis. The results showed a significant reduction in luminosity after the staining process in addition to an increase in the colors red and yellow (P whitening toothpastes and the changes found in slabs brushed with ordinary toothpaste. The whitening toothpastes did not outperform an ordinary toothpaste in the removal of extrinsic staining.

  4. Correlation of clinicopathologic parameters and immunohistochemical features of triple-negative invasive lobular carcinoma.

    Science.gov (United States)

    Harbhajanka, Aparna; Lamzabi, Ihab; Singh, Rohit I; Ghai, Ritu; Reddy, Vijaya B; Bitterman, Pincas; Gattuso, Paolo

    2014-07-01

    Invasive lobular carcinoma (ILC) is a subtype of invasive breast carcinoma. With the advent of gene profiling, breast cancer has been classified into luminal A, luminal B, HER2-overexpressing, and triple-negative carcinoma (TNC). Several studies have described TNC (ER, PR, HER2) as a surrogate for basal-like breast carcinoma. However, there is sparse literature on triple-negative lobular carcinoma (TNLC), as most of them show hormone receptor expression. The aim of this study was to investigate the correlation of clinicopathologic parameters of TNLC that has been demonstrated in invasive ductal carcinoma. Clinicopathologic parameters and immunohistochemical stains for ER, PR, E-cadherin, HER2, MIB1, and fluorescent in situ hybridization for HER2 of 255 ILC cases were retrieved. In addition, immunohistochemical analysis was performed for p53, c-kit, vimentin, p16, cyclinD1, and BCL2 on 78 cases where 12 were TNC cases and 66 were non-TNC cases. Of the 255 ILC cases, 218 (85.5%) were classic and 37 (14.5%) were pleomorphic. Seventy-seven (30.1%) cases showed axillary lymph node metastasis. There were 14 of the 255 TNC cases (5.49%) that showed higher incidence in the elderly patients. Six of the 37 (16.21%) cases were pleomorphic and 8 of the 218 (3.7%) cases were classic. Positivity for vimentin was seen in 8 of the 12 cases (67.7%), CK 5 in 3 of the 12 (25%) cases, p16 in 11 of the 12 (91.6%) cases, p53 in 8 of the 12 (66.7%) cases, c-kit in 6 of the 12 (50%) cases, and cyclinD1 in 6 of the 12 cases (50%) indicating basal-like phenotype in 3 cases and nonbasal-like phenotype in 9 cases. There was no statistical significance in lymph node metastasis, tumor recurrence, and distant metastasis between TNC and non-TNC. TNLC showed distinct clinicopathologic features such as more frequently seen in the elderly, pleomorphic, larger tumor size, increased expression of vimentin, CK 5, p16, p53, and c-kit. Not all cases showed basal-like phenotype. TNLC is less frequently

  5. An in vitro screening assay for dental stain cleaning.

    Science.gov (United States)

    Wang, Changxiang; Lucas, Robert; Smith, Anthony J; Cooper, Paul R

    2017-01-09

    The present study aimed to develop an in vitro model for stain removal from natural enamel for the assessment and comparison of oral hygiene products. Bovine teeth (n = 8 per group) were ground/polished to provide flat enamel specimens and ferric-tannate deposits were precipitated onto the enamel surfaces. The ferric-tannate stained enamel specimens were brushed using an in vitro tooth-brushing simulator with slurries containing commercially available toothpaste products, dental abrasive particles, and sodium tripolyphosphate (STP) solutions of different concentrations. The colour of the enamel surfaces was measured using a spectrophotometer before and after stain application as well as after the brushing treatments. Differences in stain removal efficacy were found between the toothpastes categorised as whitening and non-whitening comprising of different types of dental abrasives (hydrated silica and alumina). A mean value of 27% for stain removal was detected for the three non-whitening toothpastes and 59% of stain removal was detected for the three whitening toothpastes after 1000 strokes. Compared with the slurry with Zeodent 113 abrasive alone, the addition of STP provided better performance for stain removal under the same brushing conditions (mean value of 62% for Zeodent 113 abrasive alone and 72% with the addition of 5% (w/w) STP after 1000 strokes). No difference was evident between the STP concentration of 5% (w/w) and 10% (w/w). The ferric-tannate/bovine enamel model reported here provides good stain retention, is rapidly and easily prepared, and is shown to be progressively and reproducibly sensitive to toothbrushing using different toothpastes and surfactant/chelating agent solutions. Importantly, it provides good discrimination between various oral hygiene products. The stain removal assay reported here has considerable potential to enable comparative assessments of different toothpaste types in terms of their cleaning capabilities.

  6. Study of P21 Expression in Oral Lichen Planus and Oral Squamous Cell Carcinoma by Immunohistochemical Technique.

    Science.gov (United States)

    Baghaei, Fahimeh; Shojaei, Setareh; Afshar-Moghaddam, Noushin; Zargaran, Massoumeh; Rastin, Verisheh; Nasr, Mohsen; Moghimbeigi, Abbas

    2015-09-01

    Lichen planus is a mucocutaneous disease that is relatively common in middle aged individuals. Some studies have shown that oral lichen planus has a potential to progress to squamous cell carcinoma.p21 is a cyclin-dependent kinase inhibitor that regulates the cell cycle, thus it acts as an inhibitor in cell proliferation. This study was aimed to evaluate and compare the immunostaining of p21 (as a proliferation inhibitory factor) in oral lichen planus (OLP) and oral squamous cell carcinoma (OSCC). In this descriptive cross-sectional study, p21expression was investigated in 24 samples of oral lichen planus (OLP), 24 samples of oral squamous cell carcinoma (OSCC) and 24 samples of oral epithelial hyperplasia (OEH) by employing immunohistochemical staining. The mean percentage of p21-positive cells in OSCC (54.5±6.6) was significantly higher than that in OLP (32.8±6.08) and OEH (9.4±3.8). Moreover, OLP samples expressed p21 significantly higher than the OEH. Kruskal Wallis test revealed a statistically significant difference between the groups regarding the intensity of staining (plichen planus to SCC. Therefore, continuous follow-up periods for OLP are recommended for diagnosis of the malignant transformations in early stages.

  7. Immunohistochemical evaluation of estrogen and progesterone receptors in peripheral and central giant cell granuloma of the jaws

    Directory of Open Access Journals (Sweden)

    Razavi SM

    2006-07-01

    Full Text Available Background and Aim: Giant cell granuloma is a relatively common benign proliferative lesion of the oral cavity. This lesion has a marked gender predilection with more prevalence in females and tendency to rapid growth and recurrence during pregnancy. The aim of this study was the evaluation of specific receptors of sex hormones in giant cell granuloma. Materials and Methods: In this cross-sectional study, twenty five cases of formalin fixed paraffin embedded giant cell granulomas were retrieved from the oral pathology archive of dental school Isfahan University of Medical Sciences. Also twenty five normal oral mucosa biopsies resected during different surgical procedures were prepared as control group. Cases were immunohistochemically stained for estrogen and progesterone receptors using the biotin-streptavidine method. Data were analyzed by SPSS package. Results: Staining for ER/PR markers were negative for the mononuclear stromal cells and multinucleated giant cells in all cases. The epithelial cells and connective tissue stromal cells of the control group were also negative for these receptors. Conclusion: Based on the results of this study, immunostaining for ER/PR was negative in all cases. These findings suggest that in most cases development and growth of this lesion is not directly related to these hormones. However further studies with more sensitive techniques are recommended.

  8. A statistical framework for analyzing hypothesized interactions between cells imaged using multispectral microscopy and multiple immunohistochemical markers

    Directory of Open Access Journals (Sweden)

    Chris J Rose

    2013-01-01

    Full Text Available Background: Multispectral microscopy and multiple staining can be used to identify cells with distinct immunohistochemical (IHC characteristics. We present here a method called hypothesized interaction distribution (HID analysis for characterizing the statistical distribution of pair-wise spatial relationships between cells with particular IHC characteristics and apply it to clinical data. Materials and Methods: We retrospectively analyzed data from a study of 26 follicular lymphoma patients in which sections were stained for CD20 and YY1. HID analysis, using leave-one-out validation, was used to assign patients to one of two groups. We tested the null hypothesis of no difference in Kaplan-Meier survival curves between the groups. Results: Shannon entropy of HIDs assigned patients to groups that had significantly different survival curves (median survival was 7.70 versus 110 months, P = 0.00750. Hypothesized interactions between pairs of cells positive for both CD20 and YY1 were associated with poor survival. Conclusions: HID analysis provides quantitative inferences about possible interactions between spatially proximal cells with particular IHC characteristics. In follicular lymphoma, HID analysis was able to distinguish between patients with poor versus good survival, and it may have diagnostic and prognostic utility in this and other diseases.

  9. In vivo effects of focused shock waves on tumor tissue visualized by fluorescence staining techniques.

    Science.gov (United States)

    Lukes, Petr; Zeman, Jan; Horak, Vratislav; Hoffer, Petr; Pouckova, Pavla; Holubova, Monika; Hosseini, S Hamid R; Akiyama, Hidenori; Sunka, Pavel; Benes, Jiri

    2015-06-01

    Shock waves can cause significant cytotoxic effects in tumor cells and tissues both in vitro and in vivo. However, understanding the mechanisms of shock wave interaction with tissues is limited. We have studied in vivo effects of focused shock waves induced in the syngeneic sarcoma tumor model using the TUNEL assay, immunohistochemical detection of caspase-3 and hematoxylin-eosin staining. Shock waves were produced by a multichannel pulsed-electrohydraulic discharge generator with a cylindrical ceramic-coated electrode. In tumors treated with shock waves, a large area of damaged tissue was detected which was clearly differentiated from intact tissue. Localization and a cone-shaped region of tissue damage visualized by TUNEL reaction apparently correlated with the conical shape and direction of shock wave propagation determined by high-speed shadowgraphy. A strong TUNEL reaction of nuclei and nucleus fragments in tissue exposed to shock waves suggested apoptosis in this destroyed tumor area. However, specificity of the TUNEL technique to apoptotic cells is ambiguous and other apoptotic markers (caspase-3) that we used in our study did not confirmed this observation. Thus, the generated fragments of nuclei gave rise to a false TUNEL reaction not associated with apoptosis. Mechanical stress from high overpressure shock wave was likely the dominant pathway of tumor damage. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Sperm viability staining in ecology and evolution: potential pitfalls

    DEFF Research Database (Denmark)

    Holman, Luke

    2009-01-01

    a number of interesting results, it has some potential pitfalls that have rarely been discussed. In the present paper, I review the major findings of ecology and evolution studies employing sperm viability staining and outline the method's principle limitations. The key problem is that the viability assay......The causes and consequences of variation in sperm quality, survival and ageing are active areas of research in ecology and evolution. In order to address these topics, many recent studies have measured sperm viability using fluorescent staining. Although sperm viability staining has produced...

  11. News from the Biological Stain Commission No. 11

    DEFF Research Database (Denmark)

    Lyon, H O; Horobin, R W

    2012-01-01

    of Regulatory Affairs, the Biological Stain Commission's International Affairs Committee presents information from the opening session of the meeting of the International Standards Organization ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test systems held on 2-4 June 2010 in Seoul, Republic......The 11th issue of News from the Biological Stain Commission (BSC) provides our first impressions of the REACH and ECHA programs. We intend to give a more thorough account of what these important programs actually mean in later editions of News from the Biological Stain Commission. Under the heading...

  12. Expanding subjectivities

    DEFF Research Database (Denmark)

    Lundgaard Andersen, Linda; Soldz, Stephen

    2012-01-01

    A major theme in recent psychoanalytic thinking concerns the use of therapist subjectivity, especially “countertransference,” in understanding patients. This thinking converges with and expands developments in qualitative research regarding the use of researcher subjectivity as a tool to understa...

  13. Myeloid sarcomas: a histologic, immunohistochemical, and cytogenetic study

    Directory of Open Access Journals (Sweden)

    Rodgers William H

    2007-10-01

    Full Text Available Abstract Context. - Myeloid sarcoma (MS is a neoplasm of immature granulocytes, monocytes, or both involving any extramedullary site. The correct diagnosis of MS is important for adequate therapy, which is often delayed because of a high misdiagnosis rate. Objective. - To evaluate the lineage differentiation of neoplastic cells in MS by immunohistochemistry, and to correlate the results with clinicopathologic findings and cytogenetic studies. Design. - Histologic and immunohistochemical examinations were performed on formalin-fixed paraffin-embedded tissue samples from 13 cases of MS. They were classified according to the World Health Organization criteria. Chromosomal analysis data were available in 11 cases. Clinical, pathological, and cytogenetic findings were analyzed. Results. - The study included six male and seven female patients with an age range of 25 to 72 years (mean, 49.3 years and a male to female ratio of 1:1.2. MS de novo occurred in 4/13 (31% of cases examined. The most sensitive immunohistochemical markers were CD43 and lysozyme present in all cases with MS (13/13, 100%. All de novo MS showed a normal karyotype, monoblastic differentiation, and lack of CD34. The most common chromosomal abnormalities in MS associated with a hematopoietic disorder were trisomy 8 and inv(16 (2/11, 18%. Conclusion. - An immunohistochemical panel including CD43, lysozyme, myeloperoxidase (MPO, CD68 (or CD163, CD117, CD3 and CD20 can successfully identify the vast majority of MS variants in formalin-fixed paraffin-embedded tissue sections. The present report expands the spectrum of our knowledge showing that de novo MS has frequent monoblastic differentiation and frequently carries a normal karyotype.

  14. Clinicopathologic, Immunohistochemical, and Molecular Features of Histiocytoid Sweet Syndrome.

    Science.gov (United States)

    Alegría-Landa, Victoria; Rodríguez-Pinilla, Socorro María; Santos-Briz, Angel; Rodríguez-Peralto, José Luis; Alegre, Victor; Cerroni, Lorenzo; Kutzner, Heinz; Requena, Luis

    2017-07-01

    Histiocytoid Sweet syndrome is a rare histopathologic variant of Sweet syndrome. The nature of the histiocytoid infiltrate has generated considerable controversy in the literature. The main goal of this study was to conduct a comprehensive overview of the immunohistochemical phenotype of the infiltrate in histiocytoid Sweet syndrome. We also analyze whether this variant of Sweet syndrome is more frequently associated with hematologic malignancies than classic Sweet syndrome. This is a retrospective case series study of the clinicopathologic, immunohistochemical, and molecular features of 33 patients with a clinicopathologic diagnosis of histiocytoid Sweet syndrome was conducted in the dermatology departments of 5 university hospitals and a private laboratory of dermatopathology. The clinical, histopathological, immunohistochemical, and follow-up features of 33 patients with histiocytoid Sweet syndrome were analyzed. In some cases, cytogenetic studies of the dermal infiltrate were also performed. We compare our findings with those of the literature. The dermal infiltrate from the 33 study patients (20 female; median age, 49 years; age range, 5-93 years; and 13 male; median age, 42 years; age range, 4-76 years) was mainly composed of myeloperoxidase-positive immature myelomonocytic cells with histiocytoid morphology. No cytogenetic anomalies were found in the infiltrate except in 1 case in which neoplastic cells of chronic myelogenous leukemia were intermingled with the cells of histiocytoid Sweet syndrome. Authentic histiocytes were also found in most cases, with a mature immunoprofile, but they appeared to be a minor component of the infiltrate. Histiocytoid Sweet syndrome was not more frequently related with hematologic malignancies than classic neutrophilic Sweet syndrome. The dermal infiltrate of cutaneous lesions of histiocytoid Sweet syndrome is composed mostly of immature cells of myeloid lineage. This infiltrate should not be interpreted as leukemia cutis.

  15. Melan-a-positive "pseudomelanocytic nests": a pitfall in the histopathologic and immunohistochemical diagnosis of pigmented lesions on sun-damaged skin.

    Science.gov (United States)

    Beltraminelli, Helmut; Shabrawi-Caelen, Laila El; Kerl, Helmut; Cerroni, Lorenzo

    2009-05-01

    We encountered recently 3 cases with a histopathologic diagnosis of melanoma in situ on sun-damaged skin (male = 2, female = 1; median age: 59 years; range: 52-60 years). The diagnosis was based mainly on the finding of actinic elastosis in the dermis and increased number of melanocytes in the epidermis and was confirmed by strong positivity for Melan-A in single cells and in small nests ("pseudomelanocytic nests"), located at the dermoepidermal junction. Indeed, examination of slides stained with hematoxylin and eosin revealed the presence of marked hyperpigmentation and small nests of partially pigmented cells at the dermoepidermal junction, positive for Melan-A. The histologic and especially the immunohistochemical features were indistinguishable from those of melanoma in situ on chronic sun-damaged skin. In addition, a variably dense lichenoid inflammation was present. Clinicopathologic correlation, however, showed, in all patients, the presence of a lichenoid dermatitis (phototoxic reaction, 1 case; lichen planus pigmentosus, 1 case; and pigmented lichenoid keratosis, 1 case). Our cases clearly show the histopathologic pitfalls represented by lichenoid reactions on chronic sun-damaged skin. Immunohistochemical investigations, especially if performed with Melan-A alone, may lead to confusing and potentially disastrous results. The unexpected staining pattern of Melan-A in cases like ours raises concern about the utility of this antibody in the setting of a lichenoid tissue reaction on chronic sun-damaged skin. It should be underlined that pigmented lesions represent a paradigmatic example of how immunohistochemical results should be interpreted carefully and always in conjunction with histologic and clinical features.

  16. Calcifying Odontogenic Cyst with Complex Odontoma: Histological and Immunohistochemical Features

    Directory of Open Access Journals (Sweden)

    Mohsen Merati

    2012-09-01

    Full Text Available The calcifying odontogenic cyst (COC is a rare odontogenic cyst. Only 2% of all odontogenic cysts and tumors are COC. COC associated with odontoma (COCaO reported in 24% of COCs. COCaO presents a greater incidence in female, with a ratio of 2 to 1. The highest incidence of COCaO occurs during the second decade with a mean age of 16 years, most frequently occurring in the maxilla (61.5%. Here, we describe a classic case of COCaO of the maxillary incisor-canine region in 17-year-old girl, and discuss the clinicopathological features and immunohistochemical finding of this tumor.

  17. Calcifying Odontogenic Cyst with Complex Odontoma: Histological and Immunohistochemical Features

    Directory of Open Access Journals (Sweden)

    Nooshin Mohtasham

    2013-01-01

    Full Text Available The calcifying odontogenic cyst (COC is a rare odontogenic cyst. Only 2% of all odontogenic cysts and tumors are COC. COC associated with odontoma (COCaO reported in 24% of COCs. COCaO presents a greater incidence in female, with a ratio of 2 to 1. The highest incidence of COCaO occurs during the second decade with a mean age of 16 years, most frequently occurring in the maxilla (61.5%. Here, we describe a classic case of COCaO of the maxillary incisor-canine region in 17-year-old girl, and discuss the clinicopathological features and immunohistochemical finding of this tumor.

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

    Directory of Open Access Journals (Sweden)

    Sanjay D Deshmukh

    2011-01-01

    Full Text Available 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.

  19. Immunohistochemical studies of the periodontal membrane in primary teeth

    DEFF Research Database (Denmark)

    Bille, Marie-Louise Bastholm; Nolting, Dorrit; Kjær, Inger

    2009-01-01

    Objectives. To describe the periodontal membrane of human primary teeth immunohistochemically, while focusing on the epithelial layer of Malassez, fibers, and peripheral nerves, and to compare the findings with those of a previous study of human permanent teeth. Material and methods. Nineteen human...... could be identical to those in regions with no resorption. Conclusion. In regions without resorption, spatial organization of the periodontal membrane of primary teeth was similar to that of permanent teeth, although the number and distribution of epithelial cells and fibers differed. In regions...

  20. Comparison between Giemsa and Van Geison stains in ...

    African Journals Online (AJOL)

    Rukevwe S. Abraka

    2016-09-14

    Sep 14, 2016 ... After fixation of specimens, the cut- up was done, specimens were put in cassettes ... Rinse in stock solution of acetic acid. 2. Stain in giemsa .... analysis of tendon collagen using two-dimensional polarized light microscopy.

  1. Dye purity and dye standardization for biological staining

    DEFF Research Database (Denmark)

    Lyon, H O

    2002-01-01

    This review starts with a short discussion of what is meant by a pure dye and a brief description of how dyes are produced. A listing of the types of impurities encountered in dyes is made, followed by technical investigations and assessments of dyes and their impurities including methods...... for separating, identifying and assaying dye components. In the second part of the review, descriptions are given of the standardized staining method approach using standard staining methods for assessing stains, and practical responses to stain impurity including commercial quality control, third-party quality...... control and standardization of reagents, protocols and documentation. Finally, reference is made to the current state of affairs in the dye field....

  2. Spectral analysis of blood stains at the crime scene

    NARCIS (Netherlands)

    Edelman, G.J.

    2014-01-01

    In this thesis, we propose the use of several optical techniques for the detection, identification, and age estimation of blood stains. We explore the visible, near infrared, and mid infrared wavelength range for this purpose.

  3. The effect of decalcifying solutions on hemosiderin staining.

    Science.gov (United States)

    Byard, Roger W; Bellis, Maria

    2010-09-01

    To determine whether routine decalcification may reduce the amount of stainable iron that is visible on tissue sections, samples of liver and lung tissue with excessive iron stores were placed in three standard decalcifying solutions (i) formic acid [33%], formaldehyde [4%], and NaCl [0.85%]; (ii) formic acid [30%], formaldehyde [4%], and water; and (iii) nitric acid [5%] for 24, 48, 72, and 96 h. After exposure to the decalcifying solutions, the tissues were stained with Perls stain. The slides were examined blind and the intensity of iron staining was scored semiquantitatively from 0 to 3+. The trend in all samples over the course of the experiment (96 h) was for reduction in the intensity of hemosiderin staining. As the amount of stainable hemosiderin in tissues may be significantly altered by decalcification, the absence of hemosiderin in tissues adjacent to a fracture site does not necessarily indicate that the injury was acute. © 2010 American Academy of Forensic Sciences.

  4. Relationship between apoptosis and immunohistochemical staining for proliferating cell nuclear antigen and Ki-67 in non-small cell lung cancer.

    Science.gov (United States)

    Ishiwata, N; Jinn, Y; Tsukada, Y; Inase, N; Ichioka, M; Yoshizawa, Y

    2000-01-01

    The relationship between apoptosis and cellular proliferative activity in human non-small cell lung cancer (25 cases) was investigated using the in situ DNA nick-end labeling method and immunohistochemistry for both proliferating cell nuclear antigen (PCNA) and Ki-67 antigen. Comparison of the distribution of Ki-67-positive cells to that of apoptotic cells shows an inverse correlation in semi-serial sections. The PCNA labeling rates were closely correlated with Ki-67 labeling rates in non-small cell lung cancer. It was concluded that the immunostainings of PCNA and Ki-67 were almost equally valuable for assessing the proliferative activity in paraffin-embedded tissue from non-small cell lung cancer and that apoptosis may be related with the cell cycle in this cancer.

  5. Immunohistochemical staining for p16 and BRAFV600E is useful to distinguish between sporadic and hereditary (Lynch syndrome-related) microsatellite instable colorectal carcinomas.

    Science.gov (United States)

    Boissière-Michot, Florence; Frugier, Hélène; Ho-Pun-Cheung, Alexandre; Lopez-Crapez, Evelyne; Duffour, Jacqueline; Bibeau, Frédéric

    2016-08-01

    DNA mismatch repair (MMR) protein analysis by immunohistochemistry (IHC) can identify colorectal cancer (CRC) with microsatellite instability (MSI). As MLH1-deficient CRC can be hereditary or sporadic, markers to distinguish between them are needed. MLH1 promoter methylation assay is the reference method; however, sometimes, it is challenging on formalin-fixed paraffin-embedded tissue samples. We assessed by IHC the expression of BRAFV600E, p16, MGMT, and CDX2 in 55 MLH1-deficient MSI CRC samples (of which 8 had a germline MLH1 mutation) to determine whether this panel differentiates between sporadic and hereditary CRCs. We also analyzed MLH1 promoter methylation by methylation-specific PCR and pyrosequencing and BRAF status by genotyping. None of the hereditary CRCs showed MLH1 methylation, BRAF mutation, BRAFV600E-positive immunostaining, or loss of p16 expression. We detected MLH1 promoter methylation in 67 % and a BRAF mutation in 42 % of CRC, all showing MLH1 promoter methylation. BRAFV600E IHC and BRAF genotyping gave concordant results in all but two samples. Loss of expression of p16 was found in 30 % of CRC with methylation of the MLH1 promoter, but its expression was retained in all non-methylated and part of MLH1-methylated tumors (100 % specificity, 30 % sensitivity). CDX2 and MGMT expression was not associated with MLH1 status. Thus, BRAFV600E and p16 IHC may help in differentiating sporadic from hereditary MLH1-deficient CRC with MSI. Specifically, p16 IHC might be used as a surrogate marker for MLH1 promoter methylation, because all p16-negative CRCs displayed MLH1 methylation, whereas hereditary CRCs were all p16-positive.

  6. News from the Biological Stain Commission, No. 17

    DEFF Research Database (Denmark)

    Lyon, H O

    2016-01-01

    In the 17(th) issue of News from the Biological Stain Commission (BSC) under the heading of Regulatory affairs, the Biological Stain Commission's International Affairs Committee presents information from the 20(th) meeting of ISO/TC 212 Clinical laboratory testing and in vitro diagnostic test...... systems held on October 15 - 17, 2014 in Toronto, Canada, and from the 29(th) meeting of CEN/TC 140 In vitro diagnostic medical devices held on February 3, 2015 in Berlin, Germany....

  7. News from the Biological Stain Commission no. 15

    DEFF Research Database (Denmark)

    Lyon, H O; Horobin, R W

    2014-01-01

    In the 15(th) issue of News from the Biological Stain Commission (BSC), under the heading of Regulatory affairs, the Biological Stain Commission's International Affairs Committee presents information from the plenary meetings of the International Standards Organization ISO/TC 212 Clinical laborat...... laboratory testing and in vitro diagnostic test systems held on August 22-24, 2012 in Berlin, Germany. An additional discussion of the use of food dyes in India also is included....

  8. LMP2, a novel immunohistochemical marker to distinguish renal oncocytoma from the eosinophilic variant of chromophobe renal cell carcinoma.

    Science.gov (United States)

    Zheng, Gang; Chaux, Alcides; Sharma, Rajni; Netto, George; Caturegli, Patrizio

    2013-02-01

    LMP2 is a subunit of the immunoproteasome that is overexpressed in oncocytic lesions of the thyroid gland. This study was designed to assess the expression profile and diagnostic utility of LMP2 in two renal oncocytic tumors that share similar morphologic features but have different clinical outcomes: renal oncocytoma (RO) and the eosinophilic variant of chromophobe renal cell carcinoma (CHRCC-EO). A total of 56 RO, 38 classic CHRCC, and 7 CHRCC-EO cases, as well 84 normal kidney controls, were selected from the Johns Hopkins surgical pathology archive and stained for LMP2 using a standard immunohistochemical protocol. Sections were scored for cellular location (nuclear versus cytosolic), intensity (from 0 to 3), and percent of area involved (from 0 to 100%), and an H score was calculated multiplying the intensity by the extent of the staining signal. The cytoplasmic expression of LMP2 was similar among the renal lesions, being present in 44 of 56 (79%) ROs, 27 of 38 (71%) CHRCCs, and 7 of 7 (100%) CHRCC-EO cases. The nuclear expression of LMP2, however, was more informative. All CHRCC-EO cases (7 of 7, 100%) strongly showed nuclear LMP2 staining, as opposed to only 2 of 56 (4%, P<0.0001) ROs and 9 of 38 (24%, P=0.0001) classic CHRCCs. These results suggest that the nuclear LMP2 expression can be used in clinical scenarios where histological distinction between RO and CHRCC-EO remains challenging. Copyright © 2012 Elsevier Inc. All rights reserved.

  9. Gingival advanced glycation end-products in diabetes mellitus-associated chronic periodontitis: an immunohistochemical study.

    Science.gov (United States)

    Zizzi, A; Tirabassi, G; Aspriello, S D; Piemontese, M; Rubini, C; Lucarini, G

    2013-06-01

    The accumulation of advanced glycation end-products (AGEs) seems to play an important role in the development of diabetes mellitus (DM)-associated periodontitis; however, some aspects of this issue are still scarcely known, such as the expression of AGEs in type 1 DM-associated periodontitis and the clinical factors able to affect their accumulation. This study aimed to clarify these points by evaluating the expression of AGEs in DM-associated periodontitis. Sixteen systemically and periodontally healthy subjects and 48 subjects suffering from generalized, severe, chronic periodontitis (16 with type 1 DM, 16 with type 2 DM and 16 systemically healthy subjects) were studied clinically, periodontally and metabolically. The immunohistochemical expression of AGEs in gingival tissues was also evaluated. Subjects affected with type 1 DM presented a significantly higher percentage of AGE-positive cells than did subjects affected with type 2 DM, not only in the epithelium, but also in vessels and fibroblasts. A positive and significant correlation was found between gingival expression of AGEs and length of time affected with DM both in type 1 and type 2 DM; glycated hemoglobin, lipid profile, body mass index and age did not correlate significantly with gingival AGEs in any of the classes of subjects studied. Gingival AGEs are increased in both type 1 and type 2 DM-associated periodontitis; however, the clinical parameter that determines their accumulation, and therefore their degree of influence on the development of DM-associated periodontitis, may be the duration of DM. © 2012 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  10. MANAJEMEN SARANA DAN PRASARANA PENDIDIKAN DI STAIN PAMEKASAN

    Directory of Open Access Journals (Sweden)

    M. Muchlis Solichin

    2011-07-01

    Full Text Available Mediums management and pre-mediums represent an absolute done in an higher education institute, because Mediums and premediums in education management represent the absolut condition in the effort to reach the target which is expected. Thereby, Every the education organizer have to pay attention and conscripting the mind and energy to carry out education management that is professional and fulfill Standard National Education ( SNP. This Research copes to comprehend the mediums and pre-mediums management of education in STAIN Pamekasan, because during this time of mediums and basic mediums management are not yet showing its idealitas. This research is focussed at; a How mediums and pre-mediums menegement in STAIN Pamekasan ?,and b what Factors influencing mediums and pre-mediums management in STAIN Pamekasan ?. This research uses the qualitative type by using observation, interview, and documentation method. Based the rearch done, to be expressed that the first of STAIN Pamekasan conduct mediums and pre-mediums manegement still have the centralization character of top down, either in the case of planning, organizational, observation, and assessment of mediums and pre-mediums management owned, second in some cases of STAIN Pamekasan do not yet manage the mediums and pre-mediums management because they are caused by factor is its lack of management professionalism, either when doing the planning, organizational, treatment and observation or evaluation. Based the matter above, hence, suggested that STAIN Pamekasan carry out the mediums and pre-mediums management of education professionally.

  11. Lugol staining pattern and histology of esophageal lesions.

    Science.gov (United States)

    Mori, M; Adachi, Y; Matsushima, T; Matsuda, H; Kuwano, H; Sugimachi, K

    1993-05-01

    To analyze the relationship between Lugol unstained areas and their histologic features, we applied the Lugol test to 24 specimens of resected esophagus. The staining patterns were graded into four types: grade I, hyperstaining; grade II, normal greenish brown staining; grade III, less intense staining; and grade IV, unstained. Most of the grade IV lesions were invasive carcinomas, carcinomas in situ, or severe dysplasia. The carcinomas in situ and the intraepithelial extension of the carcinomas, which were difficult to detect, were clearly shown as grade IV. On the other hand, moderate to mild dysplasia or atrophy showed grade III staining. Grade IV lesions showed well-demarcated sharp margins, whereas grade III lesions showed ill-demarcated dull margins. The grade III carcinomas, however, by the Lugol test, showed well-demarcated margins. Histologic evaluation disclosed that the staining intensity reflected well the thickness of the glycogen-containing cell layer in the lesion. The sharpness of the margin reflected the abrupt or gradual change from the glycogen-containing to non-containing cell layers. These findings suggest 1) the usefulness of the staining pattern of the Lugol test for the diagnosis of esophageal lesions such as squamous cell carcinoma and severe dysplasia, and 2) the usefulness of the Lugol test for precise delineation of the proximal resection line during surgery of esophageal carcinomas with unexpected wide extension.

  12. Low-grade fibromyxoid sarcoma: case report and immunohistochemical study.

    Science.gov (United States)

    Nichols, G E; Cooper, P H

    1994-08-01

    A case is presented of low-grade fibromyxoid sarcoma involving the arm of a 52-year-old man. Low-grade fibromyxoid sarcoma is a recently described neoplasm of the deep and subcutaneous soft tissue which demonstrates a spectrum of histologic images. The current case demonstrated the typical patterns of intermixed, sweeping bands of fibrous and myxoid tissue, homogeneous foci of fibrous and myxoid tissue, focal areas of storiforming, and concentric perivascular cuffs of slender spindle cells, all lacking the nuclear anaplasia, mitotic activity, and necrosis generally associated with sarcoma. Immunohistochemical analysis performed on paraffin-embedded sections demonstrated strong labeling of the tumor cells by anti-CD34 antibody, moderate labeling for vimentin, and rare, focal positivity for muscle-specific actin. Tumor cells were negative for markers of epithelial, muscular, neural, histiocytic, melanocytic, and vascular differentiation. The constellation of histopathologic features described in this and previous reports is characteristic of low-grade fibromyxoid sarcoma. Based on this case, it appears that the immunohistochemical features of low-grade fibromyxoid sarcoma can help to exclude many cutaneous and deep soft tissue tumors from the differential diagnosis. The findings support the interpretation that the neoplasm is essentially fibroblastic in nature.

  13. Retrospective study and immunohistochemical analysis of canine mammary sarcomas.

    Science.gov (United States)

    Dolka, Izabella; Sapierzyński, Rafał; Król, Magdalena

    2013-12-09

    Canine mammary sarcomas (CMSs) are rarely diagnosed in female dogs, which explains the scarcity of immunohistochemical findings concerning those tumors. This paper presents the results of a retrospective study into CMSs and discusses the clinical features of the analyzed tumors, the expression of intermediate filaments CK, Vim, Des and α-SMA, and the expression of p63, Ki67, ERα, PR and p53 protein. Four percent of all canine mammary tumors (CMTs) were classified as CMSs, and they represented 5.1% of malignant CMTs. The mean age at diagnosis was 11.1 ± 2.8 years. Large breed dogs were more frequently affected (38.7%). The majority of observed CMSs were fibrosarcomas (2.1%). All CMSs expressed vimentin, and higher levels of vimentin expression were noted in fibrosarcomas and osteosarcomas. Ki67 expression was significantly correlated with the grade of CMS. Our results revealed that CMSs form a heterogeneous group, therefore, immunohistochemical examinations could support differential and final diagnosis. Although this study analyzed a limited number of samples, the reported results can expand our knowledge about CMSs. Further work is required in this field.

  14. Molecular and immunohistochemical distinction of equine sarcoid from schwannoma.

    Science.gov (United States)

    Bogaert, L; Heerden, M Van; Cock, H E V De; Martens, A; Chiers, K

    2011-05-01

    Ten equine skin tumors that had been classified as schwannomas on routine histological examination were analyzed by polymerase chain reaction for bovine papillomavirus DNA. All 10 were positive for bovine papillomavirus 1 or 2, and all 10 were immunohistochemically negative for S-100 protein and strongly positive for vimentin. Nine tumors were moderately positive for laminin and 8, for smooth muscle actin. Five tumors were variably and weakly positive for type IV collagen. The lack of S-100 protein expression made Schwann cells an unlikely cell of origin, as opposed to peripheral nerve sheath tumors, which typically express S-100 protein, at least in some neoplastic cells. The immunohistochemical reactivity is consistent with myofibroblastic origin of the neoplastic cells, although smooth muscle cell or pericyte origin cannot be ruled out. These tumors represent an atypical form of equine sarcoid. Polymerase chain reaction for bovine papillomavirus and S-100 immunohistochemistry are strongly recommended for all equine skin tumors with histological characteristics typical of schwannoma or peripheral nerve sheath tumor.

  15. Immunohistochemical Expression of Vitamin-D Receptor in Oral and ...

    African Journals Online (AJOL)

    Design: Nineteen formalin fixed paraffin embedded (FFPE) blocks of OSCC and 15 Skin SCC FFPE were prepared for Abcam Rabbit polyclonal Anti- Vitamin D Receptor antibody ChIP Grade (ab3508). Nuclear brown staining was considered as positive for VDR. The Sinicrope scoring method was used to evaluate the ...

  16. Behavioral and Immunohistochemical Study of the Effects of Subchronic and Chronic Exposure to Glyphosate in Mice.

    Science.gov (United States)

    Ait Bali, Yassine; Ba-Mhamed, Saadia; Bennis, Mohamed

    2017-01-01

    Many epidemiological studies have described an adolescent-related psychiatric illness and sensorimotor deficits after Glyphosate based herbicide (GBH) exposure. GBH exposure in animal models of various ages suggests that it may be neurotoxic and could impact brain development and subsequently, behavior in adulthood. However, its neurotoxic effects on adolescent brain remain unclear and the results are limited. The present study was conducted to evaluate the neurobehavioral effects of GBH following acute, subchronic (6 weeks) and chronic (12 weeks) exposure (250 or 500 mg/kg/day) in mice treated from juvenile age until adulthood. Mice were subjected to behavioral testing with the open field (OF), the elevated plus maze, the tail suspension and Splash tests (STs). Their behaviors related to exploratory activity, anxiety and depression-like were recorded. After completion of the behavioral testing, adult mice were sacrificed and the expression of tyrosine hydroxylase (TH) in the substantia nigra pars compacta (SNc) and serotonin (5-HT) in the dorsal raphe nucleus (DRN), the basolateral amygdala (BLA) and the ventral medial prefrontal cortex (mPFC) was evaluated using immunohistochemical procedure. Our results indicate that unlike acute exposure, both subchronic and chronic exposure to GBH induced a decrease in body weight gain and locomotor activity, and an increase of anxiety and depression-like behavior levels. In addition, the immunohistochemical findings showed that only the chronic treatment induced a reduction of TH-immunoreactivity. However, both subchronic and chronic exposure produced a reduction of 5-HT-immunoreactivity in the DRN, BLA and ventral mPFC. Taken together, our data suggest that exposure to GBH from juvenile age through adulthood in mice leads to neurobehavioral changes that stem from the impairment of neuronal developmental processes.

  17. Behavioral and Immunohistochemical Study of the Effects of Subchronic and Chronic Exposure to Glyphosate in Mice

    Directory of Open Access Journals (Sweden)

    Yassine Ait Bali

    2017-08-01

    Full Text Available Many epidemiological studies have described an adolescent-related psychiatric illness and sensorimotor deficits after Glyphosate based herbicide (GBH exposure. GBH exposure in animal models of various ages suggests that it may be neurotoxic and could impact brain development and subsequently, behavior in adulthood. However, its neurotoxic effects on adolescent brain remain unclear and the results are limited. The present study was conducted to evaluate the neurobehavioral effects of GBH following acute, subchronic (6 weeks and chronic (12 weeks exposure (250 or 500 mg/kg/day in mice treated from juvenile age until adulthood. Mice were subjected to behavioral testing with the open field (OF, the elevated plus maze, the tail suspension and Splash tests (STs. Their behaviors related to exploratory activity, anxiety and depression-like were recorded. After completion of the behavioral testing, adult mice were sacrificed and the expression of tyrosine hydroxylase (TH in the substantia nigra pars compacta (SNc and serotonin (5-HT in the dorsal raphe nucleus (DRN, the basolateral amygdala (BLA and the ventral medial prefrontal cortex (mPFC was evaluated using immunohistochemical procedure. Our results indicate that unlike acute exposure, both subchronic and chronic exposure to GBH induced a decrease in body weight gain and locomotor activity, and an increase of anxiety and depression-like behavior levels. In addition, the immunohistochemical findings showed that only the chronic treatment induced a reduction of TH-immunoreactivity. However, both subchronic and chronic exposure produced a reduction of 5-HT-immunoreactivity in the DRN, BLA and ventral mPFC. Taken together, our data suggest that exposure to GBH from juvenile age through adulthood in mice leads to neurobehavioral changes that stem from the impairment of neuronal developmental processes.

  18. Behavioral and Immunohistochemical Study of the Effects of Subchronic and Chronic Exposure to Glyphosate in Mice

    Science.gov (United States)

    Ait Bali, Yassine; Ba-Mhamed, Saadia; Bennis, Mohamed

    2017-01-01

    Many epidemiological studies have described an adolescent-related psychiatric illness and sensorimotor deficits after Glyphosate based herbicide (GBH) exposure. GBH exposure in animal models of various ages suggests that it may be neurotoxic and could impact brain development and subsequently, behavior in adulthood. However, its neurotoxic effects on adolescent brain remain unclear and the results are limited. The present study was conducted to evaluate the neurobehavioral effects of GBH following acute, subchronic (6 weeks) and chronic (12 weeks) exposure (250 or 500 mg/kg/day) in mice treated from juvenile age until adulthood. Mice were subjected to behavioral testing with the open field (OF), the elevated plus maze, the tail suspension and Splash tests (STs). Their behaviors related to exploratory activity, anxiety and depression-like were recorded. After completion of the behavioral testing, adult mice were sacrificed and the expression of tyrosine hydroxylase (TH) in the substantia nigra pars compacta (SNc) and serotonin (5-HT) in the dorsal raphe nucleus (DRN), the basolateral amygdala (BLA) and the ventral medial prefrontal cortex (mPFC) was evaluated using immunohistochemical procedure. Our results indicate that unlike acute exposure, both subchronic and chronic exposure to GBH induced a decrease in body weight gain and locomotor activity, and an increase of anxiety and depression-like behavior levels. In addition, the immunohistochemical findings showed that only the chronic treatment induced a reduction of TH-immunoreactivity. However, both subchronic and chronic exposure produced a reduction of 5-HT-immunoreactivity in the DRN, BLA and ventral mPFC. Taken together, our data suggest that exposure to GBH from juvenile age through adulthood in mice leads to neurobehavioral changes that stem from the impairment of neuronal developmental processes. PMID:28848410

  19. Use of microwave oven improves morphology and staining of cryostat sections.

    Science.gov (United States)

    Kennedy, A; Foulis, A K

    1989-01-01

    The quality of microscopic image of cryostat sections that had been subjected to microwave assisted fixation was compared with that resulting from conventional air drying of the sections. The role of microwaves in producing rapid special stains on cryostat sections was also assessed. Methods used permitted stains such as periodic acid Schiff, alcian blue, Gordon and Sweets's reticulin, Masson Fontana, Elastica, Prussian blue and Van Gieson to be performed within three minutes of cutting a cryostat section. The cytological detail of nuclei was much clearer using the microwave technique, allowing more accurate determination of cell type. The microwave oven seems to have major potential in improving the diagnostic accuracy of surgical frozen sections. Images Fig 1 Fig 2 Fig 3 Fig 4 Fig 5 PMID:2466053

  20. Mercury-selenium interactions in relation to histochemical staining of mercury in the rat liver

    DEFF Research Database (Denmark)

    Baatrup, E; Thorlacius-Ussing, O; Nielsen, H L

    1989-01-01

    micrograms of Se g-1 body weight as sodium [75Se]selenite. All the rats were killed 1 h later. Homogenized liver samples were prepared for mercury analysis by two different methods: alkaline digestion and ultrasonic disintegration. Quantitative chemical analysis based on benzene extraction......Selenium has been suggested to enhance the histochemical staining of mercury when sections of tissue are subjected to the silver-enhancement method. In the present study, histochemical staining patterns of mercury in tissue sections of rat livers were compared with the actual content of organic...... and inorganic Hg in the livers, in both the presence and the absence of Se. Rats were injected intravenously with 5 micrograms of Hg g-1 body weight as methyl [203Hg] mercury chloride (MeHg) or as [203Hg]mercuric chloride (Hg2+). After 2 h, half the rats received an additional intraperitoneal injection of 2...

  1. The value of intraoperative Gram stain in revision spine surgery.

    Science.gov (United States)

    Shifflett, Grant D; Nwachukwu, Benedict U; Bjerke-Kroll, Benjamin T; Kueper, Janina; Koltsov, Jayme B; Sama, Andrew A; Girardi, Federico P; Cammisa, Frank P; Hughes, Alexander P

    2015-10-01

    Intraoperative cultures and Gram stains are often obtained in cases of revision spine surgery even when clinical signs of infection are not present. The clinical utility and cost-effectiveness of this behavior remain unproven. The aim was to evaluate the clinical utility and cost-effectiveness of routine intraoperative Gram stains in revision spine surgery. This was a retrospective clinical review performed at an academic center in an urban setting. One hundred twenty-nine consecutive adult revision spine surgeries were performed. The outcome measures included intraoperative Gram stains. We retrospectively reviewed the records of 594 consecutive revision spine surgeries performed by four senior surgeons between 2008 and 2013 to identify patients who had operative cultures and Gram stains performed. All revision cases including cervical, thoracic, and lumbar fusion and non-fusion, with and without instrumentation were reviewed. One hundred twenty-nine (21.7%) patients had operative cultures obtained and were included in the study. The most common primary diagnosis code at the time of revision surgery was pseudarthrosis, which was present in 41.9% of cases (54 of 129). Infection was the primary diagnosis in 10.1% (13 of 129) of cases. Operative cultures were obtained in 129 of 595 (21.7%) cases, and 47.3% (61 of 129) were positive. Gram stains were performed in 98 of 129 (76.0%) cases and were positive in 5 of 98 (5.1%) cases. Overall, there was no correlation between revision diagnosis and whether or not a Gram stain was obtained (p=.697). Patients with a history of prior instrumentation were more likely to have a positive Gram stain (pstaining was found to have a sensitivity of 10.9% (confidence interval [CI] 3.9%-23.6%) and specificity of 100% (CI 93.1%-100%). The positive and negative predictive values were 100% (CI 48.0%-100%) and 57.3% (CI 45.2%-66.2%), respectively. Kappa coefficient was calculated to be 0.1172 (CI 0.0194-0.2151). The cost per discrepant

  2. Adult immunohistochemical markers fail to detect intratubular germ cell neoplasia in prepubertal boys with cryptorchidism.

    Science.gov (United States)

    Kvist, Kolja; Clasen-Linde, Erik; Cortes, Dina; Petersen, Bodil Laub; Thorup, Jorgen

    2014-04-01

    Intratubular germ cell neoplasia is a precursor to testicular germ cell cancer. The condition is characterized by large germ cells with large nuclei with a hyperchromatic, coarse chromatin pattern, large prominent nucleoli and abundant pale cytoplasm. In prepubertal boys these cells are located centrally and peripherally mixed with normal cells in the seminiferous tubules. We evaluated the impact of adult intratubular germ cell neoplasia marking immunohistochemistry in screening for intratubular germ cell neoplasia in boys with cryptorchidism. Histology sections of 236 testicular biopsies were retrieved from 170 boys 1 month to 15 years old operated on for cryptorchidism (excluding disorders of sex development). Specimens were incubated with primary antibodies, including anti-placental-like alkaline phosphatase, anti-Oct3/4, anti-C-kit and anti-D2-40 receptor. A 1-year, 1-month-old boy had intratubular germ cell neoplasia and all positive markers. The prevalence of placental-like alkaline phosphatase positive staining of germ cells in testicular biopsies was 98% in boys younger than 1 year, 82% in those 1 to less than 2 years old, 74% in those 2 to less than 3 years old and 60% in those 3 to 15 years. Similarly the prevalence of C-kit positive staining was 71% in boys younger than 1 year, 49% in those 1 to less than 2 years, 16% in those 2 to less than 3 years and 34% in those 3 to 15 years. Placental-like alkaline phosphatase negative germ cells did not express any of the other described antigens. In none of the 116 testes from boys older than 1 year and 7 months were any Oct3/4 or D2-40 positive germ cells identified. Up to that age 33% and 8% of biopsies were Oct3/4 and D2-40 positive, respectively. Adult intratubular germ cell neoplasia/cancer immunohistochemical markers cannot be used alone for intratubular germ cell neoplasia screening in male infants with cryptorchidism because positive immunohistochemistry is commonly seen within this age group, when most

  3. Comparison of Meldola's Blue Staining and Hatching Assay with Potato Root Diffusate for Assessment of Globodera sp. Egg Viability.

    Science.gov (United States)

    Kroese, Duncan; Zasada, Inga A; Ingham, Russell E

    2011-09-01

    Laboratory-based methods to test egg viability include staining with Meldola's Blue and/or juvenile (J2) hatching assays using potato root diffusate (PRD). These two methods have not been tested under identical conditions to directly compare their assessments of Globodera egg viability. Using two bioassay strategies, cysts from a Globodera sp. population found in Oregon were subjected to both viability assessment methods. In strategy one, intact cysts were first stained with Meldola's Blue (primary staining) and eggs were then transferred to PRD (secondary hatching). In the second strategy, intact cysts were exposed to PRD (primary hatching) and then unhatched eggs were transferred to Meldola's Blue (secondary staining). Two different cohorts of cysts were evaluated using these experimental strategies: cohort 1 was comprised of cysts produced on potato in the greenhouse that exhibited low hatch when exposed to PRD and cohort 2 consisted of field-collected cysts whose eggs yielded significant hatch when exposed to PRD. Percentage viability was calculated and is expressed as the number of hatched J2 or unstained eggs/total number of eggs within a cyst. With field-produced cysts, primary staining with Meldola's Blue and hatching with PRD produced similar viability estimates, with averages of 74.9% and 76.3%, respectively. In contrast, with greenhouse-produced cysts the two methods yielded much lower and unequal estimates 32.4% to 2.2%, respectively for primary hatching and staining methods. In addition, J2 hatch from unstained (viable) greenhouse-produced eggs was 13.7% after secondary exposure to PRD compared to 61.5% for field-produced eggs. The majority of eggs remaining unhatched after primary exposure to PRD (> 87%) stained with Meldola's Blue regardless of cyst cohort. Staining with Meldola's Blue provided a conservative assessment of egg viability compared to hatch assay with PRD regardless of diapause.

  4. Cellular and Subcellular Immunohistochemical Localization and Quantification of Cadmium Ions in Wheat (Triticum aestivum.

    Directory of Open Access Journals (Sweden)

    Wei Gao

    Full Text Available The distribution of metallic ions in plant tissues is associated with their toxicity and is important for understanding mechanisms of toxicity tolerance. A quantitative histochemical method can help advance knowledge of cellular and subcellular localization and distribution of heavy metals in plant tissues. An immunohistochemical (IHC imaging method for cadmium ions (Cd2+ was developed for the first time for the wheat Triticum aestivum grown in Cd2+-fortified soils. Also, 1-(4-Isothiocyanobenzyl-ethylenediamine-N,N,N,N-tetraacetic acid (ITCB-EDTA was used to chelate the mobile Cd2+. The ITCB-EDTA/Cd2+ complex was fixed with proteins in situ via the isothiocyano group. A new Cd2+-EDTA specific monoclonal antibody, 4F3B6D9A1, was used to locate the Cd2+-EDTA protein complex. After staining, the fluorescence intensities of sections of Cd2+-positive roots were compared with those of Cd2+-negative roots under a laser confocal scanning microscope, and the location of colloidal gold particles was determined with a transmission electron microscope. The results enable quantification of the Cd2+ content in plant tissues and illustrate Cd2+ translocation and cellular and subcellular responses of T. aestivum to Cd2+ stress. Compared to the conventional metal-S coprecipitation histochemical method, this new IHC method is quantitative, more specific and has less background interference. The subcellular location of Cd2+ was also confirmed with energy-dispersive X-ray microanalysis. The IHC method is suitable for locating and quantifying Cd2+ in plant tissues and can be extended to other heavy metallic ions.

  5. Immunohistochemical expression of HER-1 and HER-2 in extrahepatic biliary carcinoma.

    Science.gov (United States)

    Ogo, Yasumasa; Nio, Yoshinori; Yano, Seiji; Toga, Tomoko; Koike, Makoto; Hashimoto, Koji; Itakura, Masayuki; Maruyama, Riruke

    2006-01-01

    The clinicopathological significance of HER-1- and HER-2-overexpressions (OE) (HercepTest score 2+ or 3+) in biliary cancer and their relationship to the efficacy of adjuvant chemotherapy (ACT) were assessed. In 72 biliary cancer (28 gallbladder and 44 bile duct cancer), HER-1 and HER-2 were stained immunohistochemically in formalin-fixed, paraffin-embedded specimens. The ACT included uracil and tegafur (UFT)-based chemotherapies. Out of the 72 cancer, OE was observed in 31 specimens (43%) for HER-1 and 47 (65%) for HER-2. However, their OEs were not correlated with each other. HER-2-OE was inversely correlated with the clinical stage (p=0.0482). HER-1-OE was correlated with distant metastasis (p=0.0263), but not with the clinical stage. Neither the OE of HER-1 or HER-2, nor their co-expression, showed any significant effect in term of patient survival. In the HER-1-OE (-) patients, the survival rate of the ACT group was significantly higher than that of the surgery-alone (SA) group (p=0.0423), but in the HER-1-OE (+) patients, there was no statistical difference in survival rate between the ACT and the SA group. In contrast, HER-2-OE had no significant effect on the efficacy of ACT. Multivariate analysis also demonstrated that the histological grade and ACT were significant variables, but T, N, M and HER-1 and HER-2 were not significant variables. In conclusion, neither HER-1-OE or HER-2-OE were prognostic factors of the biliary cancer. However, HER-1-OE may be a useful marker for the indication of ACT.

  6. TSE strain differentiation in mice by immunohistochemical PrP(Sc) profiles and triplex Western blot.

    Science.gov (United States)

    van Keulen, Lucien J M; Langeveld, Jan P M; Dolstra, Corry H; Jacobs, Jorg; Bossers, Alex; van Zijderveld, Fred G

    2015-10-01

    TSE strains are routinely identified by their incubation period and vacuolation profile in the brain after intracerebral inoculation and serial passaging in inbred mouse lines. There are some major drawbacks to this method that are related to the variation in vacuolation that exists in the brains of mice infected with the same TSE strain and to variation between observers and laboratories in scoring vacuolation and determining the final incubation period. We investigated the potential of PrP(Sc) immunohistochemistry and triplex Western blotting as possible alternative methods to differentiate between TSE strains. TSE reference strains ME7, 87A/87V, 22A/22C, 79A/79V and 301C/301V were intracerebrally inoculated in RIII or VM inbred mice that differ in their PrP genotype. Immunohistochemical PrP(Sc) profiles were drawn up by scanning light microscopy both on coronal and sagittal sections. On the basis of the localization of PrP(Sc) in the cerebral cortex, hippocampus, and cerebellar cortex and the overall type of PrP(Sc) staining, all TSE strains could be well differentiated from each other through their typical strain dependent characteristics. In addition, Western blot showed that the combination of glycosylation profile and 12B2 epitope content of PrP(Sc) allowed to distinguish between all reference strains except for ME7 and 22A in VM mice. TSE strains in mice can be identified on the basis of their PrP(Sc) profile alone. The potential to identify TSE strains in ruminants with these PrP(Sc) profiles after a single primary passage in mice will be the topic of future studies. © 2014 British Neuropathological Society.

  7. Immunohistochemical Expression of Cyclooxygenase-2 in Urinary Bladder Transitional Cell Carcinomas

    Directory of Open Access Journals (Sweden)

    F Niki

    2012-07-01

    Full Text Available Background: Transitional Cell Carcinoma (TCC is the most common type of urinary bladder cancer. Cyclooxygenase-2 (COX-2, a key enzyme in prostaglandins biosynthesis, has been introduced as a new candidate for targeted therapy in this cancer. In this study, we investigated the expression of COX-2 in urinary bladder TCCs and its relationship with clinicopathological parameters such as tumor grade and stage. Methods: This cross-sectional study was performed in the Pathology department of Sina Hospital in Tehran, Iran during 2006-2011. Pathology reports of patients with definite diagnosis of urinary bladder TCCs who had undergone Transurethral Resection (TUR were reviewed and 40 cases were selected. Subsequently, COX-2 expression was assessed immunohistochemically by the examination of paraffin embedded tissue blocks. Staining in more than 5% of tumor cells was considered as positive expression. Results: COX-2 was expressed in 52.5% of the patients. High-grade tumors revealed a higher (87.5% COX-2 expression versus other grades of the lesions and there was a statistically significant difference in COX-2 expression between them (P<0.001. Patients age was also related to the expression of this marker (P=0.03. In contrast, this marker did not correlate with other characteristics including gender, lymphatic invasion or tumor stage. In addition, perineurial or vascular invasions were not detected in any of the patients. Conclusion: COX-2 expression was seen in more than half of our patients and it had a marked relation to tumor differentiation. Accordingly, this molecule may be a useful tumor marker in the assessment of urinary bladder cancers.

  8. Protein tyrosine nitration in chronic intramuscular parasitism: immunohistochemical evaluation of relationships between nitration, and fiber type-specific responses to infection

    Directory of Open Access Journals (Sweden)

    Ted H. Elsasser

    2011-05-01

    Full Text Available The present study was conducted to determine whether preferential muscle catabolism [psoas major (PM > rectus femoris (RF] observed during the chronic intramuscular stage of Sarcocystis cruzi infection could be associated with the pathological consequences of increased protein tyrosine nitration in fibers characteristically more metabolically active due to higher mitochondrial density. Holstein calves were assigned to control (C, or S. cruzi-infected (I groups, n=5/group. Calves were euthanized on day 63 of infection. Samples of RF and PM were prepared for metabolic fiber typing (MFT: slow oxidative, SO – Type I; fast oxidative glycolytic, FOG - Type IIa; fast glycolytic, FG – Type IIb, fiber area, and immunohistochemical localization of fast myosin heavy chain 2a and 2b, nitrotyrosine (NT, and mitochondrial Complex V ATP-synthase. MFT analysis documented that PM contained twice the number of SO fibers compared to RF (32 v 16%, P<0.002. SO and FOG fibers (Both higher in mitochondrial density than FG fibers in both PM and RF were significantly smaller in area in I calves with mean FG areas not different between C and I. Muscle NT content (Western blot of myofibrillar protein fraction increased with infection; NT was immunohistochemically localized into three distinct patterns in fibers: i sparse fiber staining, ii dense punctuate intrafiber staining, and iii pericystic staining. By image analysis, the greatest punctuate intrafiber pixel density of NT was associated with SO fibers from I calves with the NT colocalizing with mitochondrial Complex V – F1F0 ATP synthase. More fibers were positive for the colocalization in PM than RF (P<0.04. The data are consistent with the concept that fibers rich in mitochondria possessing more inherent oxidative energy capacity generate more nitrated proteins than glycolytic fibers and as such are more affected by the proinflammatory response to infections like Sarcocystosis.

  9. Immunohistochemical and histochemical characterization of intraosseous arteriovenous malformations of the jaws: analysis of 16 cases with emphasis on GLUT-1 immunophenotype.

    Science.gov (United States)

    Taleb, Reda; Koutlas, Ioannis G; Argyris, Prokopios P

    2017-08-01

    Intraosseous vascular lesions of the craniofacial region are rare and may cause diagnostic and therapeutic challenges. The purpose of this study was to characterize 16 cases of intraosseous arteriovenous malformations (AVMs) affecting the jaws. Immunohistochemical evaluation was performed using antibodies against α-smooth muscle actin (α-SMA), desmin, CD31, D2-40, and glucose transporter 1 (GLUT-1). Staining with elastic Verhoeff-Van Gieson and Masson trichrome histochemical stains was also performed. No gender predilection (female:male ratio = 1:1) was observed, with patients' mean age being 50.8 years (SD of ±13.9; range 28-71 years). Predilection for the mandible was observed (12 of 16 [75%]). Immunohistochemically, diffuse endothelial CD31 staining was noted, and α-SMA strongly highlighted smooth muscle cells and pericytes. Desmin-positive vessels were identified in 9 of 16 AVMs (56.3%). D2-40 was uniformly negative in all specimens. AVMs were negative for GLUT-1 (11 of 14 [78.6%]) except for 2 cases (2 of 14 [14.3%]) exhibiting focal limited cytoplasmic GLUT-1 immunoreactivity. One case was equivocal for GLUT-1. Masson trichrome highlighted smooth muscle cells, and elastic fibers were identified in thick-walled arteries. AVMs of the jaws generally lack expression of GLUT-1, similar to soft tissue vascular malformations. Clinicoradiographic features of intraosseous AVMs in the present study were consistent with the findings of previous studies, although mean age was higher. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. A comparison of two chlorhexidine gel brushing regimens and a conventional toothpaste brushing regimen for the development of tooth staining over a 6-week period.

    Science.gov (United States)

    Claydon, N C A; Addy, M; Adams, G; Smith, S R; Bosma, M L; North, M; Moran, J

    2006-11-01

    A single-centre, randomized single-blind parallel study was undertaken to compare staining seen with three brushing regimens and to determine subject perception of side effects such as staining and effects on taste. This 6-week parallel study used 157 volunteers who were randomized into one of three treatment groups: (i) brushing in the morning and evening with a normal dose of a 1% chlorhexidine gel, (ii) brushing with a low dose of chlorhexidine gel in the evening and a whitening dentifrice in the morning, and (iii) brushing with a standard fluoride paste in the morning and evening. Following home usage of their allocated products, the study volunteers returned after 3 and 6 weeks to record the amount of stain present. After the 6-week period, subject perception of taste and stain acceptability was determined using a questionnaire. After 6 weeks of use of the low-dose chlorhexidine gel and whitening dentifrice, significantly more stain was seen compared with the use of a standard dentifrice (Pwhitening dentifrice (P=0.0007). Approximately 30% of individuals on the low-dose chlorhexidine gel regimen found the amount of stain unacceptable and 10% noted an effect on their taste perception. The use of low dose of chlorhexidine gel at night and a whitening paste in the morning produced a significant amount of stain that 30% of subjects considered unacceptable.

  11. AutoIHC-scoring: a machine learning framework for automated Allred scoring of molecular expression in ER- and PR-stained breast cancer tissue.

    Science.gov (United States)

    Tewary, S; Arun, I; Ahmed, R; Chatterjee, S; Chakraborty, C

    2017-11-01

    In prognostic evaluation of breast cancer Immunohistochemical (IHC) markers namely, oestrogen receptor (ER) and progesterone receptor (PR) are widely used. The expert pathologist investigates qualitatively the stained tissue slide under microscope to provide the Allred score; which is clinically used for therapeutic decision making. Such qualitative judgment is time-consuming, tedious and more often suffers from interobserver variability. As a result, it leads to imprecise IHC score for ER and PR. To overcome this, there is an urgent need of developing a reliable and efficient IHC quantifier for high throughput decision making. In view of this, our study aims at developing an automated IHC profiler for quantitative assessment of ER and PR molecular expression from stained tissue images. We propose here to use CMYK colour space for positively and negatively stained cell extraction for proportion score. Also colour features are used for quantitative assessment of intensity scoring among the positively stained cells. Five different machine learning models namely artificial neural network, Naïve Bayes, K-nearest neighbours, decision tree and random forest are considered for learning the colour features using average red, green and blue pixel values of positively stained cell patches. Fifty cases of ER- and PR-stained tissues have been evaluated for validation with the expert pathologist's score. All five models perform adequately where random forest shows the best correlation with the expert's score (Pearson's correlation coefficient = 0.9192). In the proposed approach the average variation of diaminobenzidine (DAB) to nuclear area from the expert's score is found to be 7.58%, as compared to 27.83% for state-of-the-art ImmunoRatio software. © 2017 The Authors Journal of Microscopy © 2017 Royal Microscopical Society.

  12. Amylase α-1A (AMY1A): a novel immunohistochemical marker to differentiate chromophobe renal cell carcinoma from benign oncocytoma.

    Science.gov (United States)

    Jain, Sarika; Roy, Somak; Amin, Milon; Acquafondata, Marie; Yin, Ming; Laframboise, William; Bastacky, Sheldon; Pantanowitz, Liron; Dhir, Rajiv; Parwani, Anil

    2013-12-01

    Chromophobe renal cell carcinoma (ChRCC) and oncocytoma present with a perplexing overlap of morphologic and immunohistochemical features. ChRCC have deletions in the 1p21.1 region including the amylase α-1A gene (AMY1A). No such deletions are found in oncocytoma. Instead, oncocytomas shared other deletions on chromosome 1: 1p31.3, 1q25.2, and 1q44. We performed AMY1A immunostaining on 75 oncocytomas (57 tissue microarray [TMA] cores, 18 whole slides) and 54 ChRCCs (20 TMA cores, 34 whole slides). Staining was assessed using the H-score method. The intensity was graded as follows: no staining=0, weak=1, moderate=2, and strong=3. The AMY1A immunostain preferentially stained the distal tubules and collecting ducts of normal kidney. All oncocytomas (100%) expressed AMY1A with an H-score that varied from 100 to 300 (mean 205). Mild to moderate heterogeneity in staining intensity was noted within a given oncocytoma. For oncocytomas, 87% (65/75) cases had H-scores of at least 120 with a mean score of 221. Notably, the 13% (10/75) of oncocytoma cases that had an H-score of 100 were derived from the TMA. A total of 87% (47/54) of the ChRCC cases were negative for the AMY1A immunostain. Of the ChRCC cases, 4% (2/54) showed very weak cytoplasmic staining (H-score of 70 each), which was less than the lowest H-score of oncocytoma cases. All 5 cases of ChRCC, which showed an H-score of 100 or more, were referred to as eosinophilic variants of ChRCC. Three of these 5 cases showed a very nondescript, diffuse staining of the cytoplasm. Two of these 5 cases showed an H-score of 130. We think that as the staining pattern of these 2 cases is similar to that of oncocytoma, they should be put in a category of renal oncocytic neoplasms favoring oncocytoma. This result shows that AMY1A staining could be very helpful in further classifying even a subset of the eosinophilic variants of ChRCC. The difference between ChRCC and oncocytoma was statistically significant (χ test, Poncocytoma

  13. Identification and quantification of microplastics using Nile Red staining.

    Science.gov (United States)

    Shim, Won Joon; Song, Young Kyoung; Hong, Sang Hee; Jang, Mi

    2016-12-15

    We investigated the applicability of Nile Red (NR), a fluorescent dye, for microplastic analysis, and determined the optimal staining conditions. Five mg/L NR solution in n-hexane effectively stained plastics, and they were easily recognized in green fluorescence. The NR staining method was successfully applied to micro-sized polyethylene, polypropylene, polystyrene, polycarbonate, polyurethane, and poly(ethylene-vinyl acetate), except for polyvinylchloride, polyamide and polyester. The recovery rate of polyethylene (100-300μm) spiked to pretreated natural sand was 98% in the NR stating method, which was not significantly (p<0.05) different with FT-IR identification. The NR staining method was suitable for discriminating fragmented polypropylene particles from large numbers of sand particles in laboratory weathering test samples. The method is straightforward and quick for identifying and quantifying polymer particles in the laboratory controlled samples. Further studies, however, are necessary to investigate the application of NR staining to field samples with organic remnants. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Usefulness of the stool Wright's stain in the emergency department.

    Science.gov (United States)

    DuBois, D; Binder, L; Nelson, B

    1988-01-01

    A prospective study was conducted to determine if a Wright's stain of stool specimen to detect fecal leukocytes was accurate in predicting the presence of a bacterial pathogen on stool culture. Entry criteria were patient age greater than or equal to 3 months and diarrhea of greater than 1 day. The patient population was drawn from an urban county hospital emergency department on the Texas-Mexican border. A total of 69 patients were evaluated by both routine stool culture and stool Wright's stain. Twenty-three were evaluated for parasitic pathogens. There were seventeen cultures positive for bacterial pathogens and twenty-three positive Wright's stains. Bacterial isolates included Shigella, Salmonella and Campylobacter. Also detected were Giardia, Shistosoma, Blastocytis and Cryptosporidium. The sensitivity of a Wright's stain positive for fecal leukocytes for the presence of a bacterial pathogen by culture was 82%, with a specificity of 83%. These were significantly correlated with a positive culture for a bacterial pathogen (P less than .01). The predictive value of a positive result was 61%, and predictive value of a negative result was 94%, for bacterial pathogens. The Wright's stain is a useful tool for the early presumptive diagnosis of infectious bacterial diarrhea in the emergency department.

  15. SUBJECT INDEX

    Indian Academy of Sciences (India)

    Subject Index. Variation of surface electric field during geomagnetic disturbed period at Maitri, Antarctica. 1721. Geomorphology. A simple depression-filling method for raster and irregular elevation datasets. 1653. Decision Support System integrated with Geographic. Information System to target restoration actions in water-.

  16. When one plus one equals more than two - a novel stain for renal biopsies is a combination of two classical stains

    OpenAIRE

    Brodsky, Sergey V.; Albaward, Alia; Satoskar, Anjali A.; Nadasdy, Gyongyi; Nadasdy, Tibor

    2010-01-01

    Histologic evaluation of renal biopsies includes multiple ancillary stains, including Periodic acid-Schiff ’s (PAS) and Masson’s trichrome (Trichrome). Herein we report an innovative doublestain, derived from two standard stains (PAS and Trichrome). This novel stain not only has advantages of both ancestor stains, but became more distinguishable and colorful, when basement membranes stain darkviolet, whereas the interstitial collagen remains blue. This allows the pa...

  17. Immunohistochemical Analysis of the Mechanistic Target of Rapamycin and Hypoxia Signalling Pathways in Basal Cell Carcinoma and Trichoepithelioma

    Science.gov (United States)

    Brinkhuizen, Tjinta; Weijzen, Chantal A. H.; Eben, Jonathan; Thissen, Monique R.; van Marion, Ariënne M.; Lohman, Björn G.; Winnepenninckx, Véronique J. L.; Nelemans, Patty J.; van Steensel, Maurice A. M.

    2014-01-01

    Background Basal cell carcinoma (BCC) is the most common cancer in Caucasians. Trichoepithelioma (TE) is a benign neoplasm that strongly resembles BCC. Both are hair follicle (HF) tumours. HFs are hypoxic microenvironments, therefore we hypothesized that hypoxia-induced signalling pathways could be involved in BCC and TE as they are in other human malignancies. Hypoxia-inducible factor 1 (HIF1) and mechanistic/mammalian target of rapamycin (mTOR) are key players in these pathways. Objectives To determine whether HIF1/mTOR signalling is involved in BCC and TE. Methods We used immunohistochemical staining of formalin-fixed paraffin-embedded BCC (n = 45) and TE (n = 35) samples to assess activity of HIF1, mTORC1 and their most important target genes. The percentage positive tumour cells was assessed manually in a semi-quantitative manner and categorized (0%, 80%). Results Among 45 BCC and 35 TE examined, expression levels were respectively 81% and 57% (BNIP3), 73% and 75% (CAIX), 79% and 86% (GLUT1), 50% and 19% (HIF1α), 89% and 88% (pAKT), 55% and 61% (pS6), 15% and 25% (pMTOR), 44% and 63% (PHD2) and 44% and 49% (VEGF-A). CAIX, Glut1 and PHD2 expression levels were significantly higher in TE when only samples with at least 80% expression were included. Conclusions HIF and mTORC1 signalling seems active in both BCC and TE. There are no appreciable differences between the two with respect to pathway activity. At this moment immunohistochemical analyses of HIF, mTORC1 and their target genes does not provide a reliable diagnostic tool for the discrimination of BCC and TE. PMID:25181405

  18. Immunohistochemical analysis of the mechanistic target of rapamycin and hypoxia signalling pathways in basal cell carcinoma and trichoepithelioma.

    Directory of Open Access Journals (Sweden)

    Tjinta Brinkhuizen

    Full Text Available BACKGROUND: Basal cell carcinoma (BCC is the most common cancer in Caucasians. Trichoepithelioma (TE is a benign neoplasm that strongly resembles BCC. Both are hair follicle (HF tumours. HFs are hypoxic microenvironments, therefore we hypothesized that hypoxia-induced signalling pathways could be involved in BCC and TE as they are in other human malignancies. Hypoxia-inducible factor 1 (HIF1 and mechanistic/mammalian target of rapamycin (mTOR are key players in these pathways. OBJECTIVES: To determine whether HIF1/mTOR signalling is involved in BCC and TE. METHODS: We used immunohistochemical staining of formalin-fixed paraffin-embedded BCC (n = 45 and TE (n = 35 samples to assess activity of HIF1, mTORC1 and their most important target genes. The percentage positive tumour cells was assessed manually in a semi-quantitative manner and categorized (0%, 80%. RESULTS: Among 45 BCC and 35 TE examined, expression levels were respectively 81% and 57% (BNIP3, 73% and 75% (CAIX, 79% and 86% (GLUT1, 50% and 19% (HIF1α, 89% and 88% (pAKT, 55% and 61% (pS6, 15% and 25% (pMTOR, 44% and 63% (PHD2 and 44% and 49% (VEGF-A. CAIX, Glut1 and PHD2 expression levels were significantly higher in TE when only samples with at least 80% expression were included. CONCLUSIONS: HIF and mTORC1 signalling seems active in both BCC and TE. There are no appreciable differences between the two with respect to pathway activity. At this moment immunohistochemical analyses of HIF, mTORC1 and their target genes does not provide a reliable diagnostic tool for the discrimination of BCC and TE.

  19. Claudin-7 and claudin-8: immunohistochemical markers for the differential diagnosis of chromophobe renal cell carcinoma and renal oncocytoma.

    Science.gov (United States)

    Osunkoya, Adeboye O; Cohen, Cynthia; Lawson, Diane; Picken, Maria M; Amin, Mahul B; Young, Andrew N

    2009-02-01

    Claudin-7 and claudin-8 code for tight junction proteins expressed in distal nephron epithelium. In a recent oligonucleotide microarray study, we identified claudin-7 and claudin-8 as candidate markers to distinguish chromophobe renal cell carcinoma from other renal tumors, including oncocytoma. Distinction of these lesions can be difficult by light microscopy but is clinically important because chromophobe renal cell carcinoma has malignant biological potential, whereas renal oncocytoma is benign. Claudin-7 and claudin-8 expression was studied by immunohistochemistry in 11 chromophobe renal cell carcinomas and 17 oncocytomas using formalin-fixed paraffin-embedded tissue sections of tumor with adjacent nonneoplastic kidney. Steam antigen retrieval was performed before immunohistochemistry. Specificity was verified by negative control reactions without primary antibody and appropriate membranous staining patterns in positive control tissues (colon carcinoma and adjacent nonneoplastic kidney). Claudin-7 protein was expressed in a membranous pattern in 10 of 11 chromophobe renal cell carcinomas and 4 of 17 oncocytomas (P oncocytoma, 11 of 17 cases showed cytoplasmic, 4 of 17 membranous, and 2 of 17 negative reactions. In chromophobe renal cell carcinoma, 0 of 11 cases showed cytoplasmic, 3 of 11 membranous, and 8 of 11 negative reactions (P renal cell carcinomas and 1 of 17 oncocytomas (63% sensitivity, 84% specificity, 88% positive predictive value for chromophobe renal cell carcinoma). Negative claudin-7 and cytoplasmic claudin-8 were observed in 10 of 17 oncocytomas and 0 of 11 chromophobe renal cell carcinomas (59% sensitivity, 100% specificity and positive predictive value for oncocytoma). The distal nephron proteins claudin-7 and claudin-8 have potential use as immunohistochemical biomarkers in the differential diagnosis of chromophobe renal cell carcinoma and oncocytoma. Expression of claudin-7 and claudin-8 may reflect the relationship of chromophobe renal

  20. Effect of oral lactulose on clinical and immunohistochemical parameters in patients with inflammatory bowel disease: a pilot study

    Directory of Open Access Journals (Sweden)

    Manns Michael P

    2007-09-01

    Full Text Available Abstract Background The prebiotic potential of lactulose is well established and preclinical studies demonstrated a protective effect of lactulose in murine models of colitis. The aim of the present study was to investigate the clinical and histological efficacy of lactulose in patients with inflammatory bowel disease (IBD, for which probiotic therapy yielded promising results. Methods Patients were treated with standard medication alone or combined with 10 g lactulose daily as adjuvant therapy for 4 months. Clinical efficacy of treatment was assessed using clinical activity indices, a quality of life index (IBDQ, endoscopic scores, defecation frequency and monitoring corticosteroid medication. Orsomucoid, alpha1-antitrypsin and other laboratory parameters were determined. In addition, in some participants colonic biopsies were analyzed with haematoxylin-eosin staining or with antibodies against HLA-DR, CD68, IgA and CD3, and evaluated systematically. All measurements were performed both at enrolment and at the end of the trial. Results 14 patients presenting ulcerative colitis (UC and 17 patients presenting Crohn's disease (CD, most of them in a clinically active state, were enrolled in this pilot study. After 4 month no significant improvement of clinical activity index, endoscopic score or immunohistochemical parameters was observed in CD or UC patients receiving lactulose in comparison to the control group. However, significant improvement of quality of life was observed in UC patients receiving lactulose compared to the control group (p = 0.04. Conclusion The findings of the present pilot study indicate that oral lactulose has no beneficial effects in IBD patients in particular with regard to clinical activity, endoscopic score or immunohistochemical parameters. The importance of the beneficial effect of lactulose in UC patients regarding the quality of life needs further evaluation in larger controlled clinical trials. Trial registration

  1. Effects of Melatonin on the Cerebellum of Infant Rat Following Kaolin-Induced Hydrocephalus: a Histochemical and Immunohistochemical Study.

    Science.gov (United States)

    Uyanıkgil, Yiğit; Turgut, Mehmet; Baka, Meral

    2017-02-01

    Hydrocephalus is a developmental disorder causing abnormally collected cerebrospinal fluid within the cerebral ventricles. It leads to bigger skulls and many dysfunctions related to the nervous system. Here, we addressed whether exogenous melatonin administration could reverse the clinical features of kaolin-induced hydrocephalus in infantile rats. A controlled double-blinded study was conducted in 2-week-old 45 Wistar albino rats, which were divided into three groups: Group A, the control group, received intracisternal sham injection with solely the needle insertion; group B, the hydrocephalus group, was treated with isotonic NaCl after kaolin injection; and group C, the hydrocephalus + melatonin group, was given i.p. exogenous melatonin at a dose of 0.5 mg/100 g body weight after kaolin injection. Histological and immunohistochemical analyses were performed after the induction of hydrocephalus and melatonin administration. Glial fibrillary acidic protein was stained by immunohistochemical method. TUNEL method was used to define and quantitate apoptosis in the cerebellar tissues. Statistical analysis was performed by nonparametric Kruskal-Wallis H test, and once significance was determined among means, post hoc pairwise comparisons were carried out using Mann-Whitney U test. We found that melatonin administration significantly ameliorated ratio of substantia grisea area/substantia alba area in the cerebellum of infantile rats. Histologically, there was a significant reduction in the number of cerebellar apoptotic cells after the hydrocephalus induced by kaolin (P cerebellum were reversed by systemic melatonin administration in infantile rats with kaolin-induced hydrocephalus. Nevertheless, further studies are needed to suggest melatonin as a candidate protective drug in children with hydrocephalus.

  2. Parachordoma of the buttock: an immunohistochemical case study and review.

    Science.gov (United States)

    Hirokawa, M; Manabe, T; Sugihara, K

    1994-12-01

    We report a case of parachordoma occurring in the buttock of a 43-year-old man, and review 20 cases of parachordoma reported in the English literature. The tumor in our case was grossly 3 cm in dimension, solid, lobulated and grayish-white in color. Microscopically, the tumor consisted of epithelioid and spindle cells, and fibromyxoid stroma. The epithelioid cells were immunohistochemically positive for vimentin, S-100 protein, neuron-specific enolase, keratin, carcinoembryonic antigen and epithelial membrane antigen, and negative for HMB45. These findings are similar to those for chordoma rather than extraskeletal myxoid chrondrosarcoma. Although the etiopathogenesis of parachordoma remains obscure, Schwann cells or some other neuron-related cell origin are suspected.

  3. Immunohistochemical evidence of specific iris involvement in Blau syndrome.

    Science.gov (United States)

    Paparizos, Stella C; Goldstein, Debra A; Bouhenni, Rachida A; Steiner, Scott; Dunmire, Jeffrey J; Edward, Deepak P

    2012-12-01

    To evaluate iris involvement in Blau syndrome using histology and immunohistochemistry. Iridectomy specimen of a patient with treated Blau syndrome and a normal control were evaluated by light microscopy and immunohistochemistry using antibodies against CD4(+), CD8(+), HLA-DR, CD68(+), NF-κB and IL-17. Blau iris tissue demonstrated increased numbers of CD4(+) lymphocytes and CD68 negative, HLA-DR positive spindle shaped cells compared to normal iris tissue. Blau iris tissue also demonstrated elevated CD4(+)/CD8(+) ratio and IL-17 and NF-κB immunolabeling. No macrophages, epithelioid cells, or granulomas were noted in the Blau specimen. The persistent immunolocalization of inflammatory markers in an iris specimen from an aggresively treated patient with proven Blau syndrome suggests that further pathologic and immunohistochemical investigation of Blau ocular tissue is necessary to better understand the complexities of NOD2 activating mutations in the eye.

  4. Ebola Virus Disease in Pregnancy: Clinical, Histopathologic, and Immunohistochemical Findings.

    Science.gov (United States)

    Muehlenbachs, Atis; de la Rosa Vázquez, Olimpia; Bausch, Daniel G; Schafer, Ilana J; Paddock, Christopher D; Nyakio, Jean Paul; Lame, Papys; Bergeron, Eric; McCollum, Andrea M; Goldsmith, Cynthia S; Bollweg, Brigid C; Prieto, Miriam Alía; Lushima, Robert Shongo; Ilunga, Benoit Kebela; Nichol, Stuart T; Shieh, Wun-Ju; Ströher, Ute; Rollin, Pierre E; Zaki, Sherif R

    2017-01-01

    Here we describe clinicopathologic features of Ebola virus disease in pregnancy. One woman infected with Sudan virus in Gulu, Uganda, in 2000 had a stillbirth and survived, and another woman infected with Bundibugyo virus had a live birth with maternal and infant death in Isiro, the Democratic Republic of the Congo in 2012. Ebolavirus antigen was seen in the syncytiotrophoblast and placental maternal mononuclear cells by immunohistochemical analysis, and no antigen was seen in fetal placental stromal cells or fetal organs. In the Gulu case, ebolavirus antigen localized to malarial parasite pigment-laden macrophages. These data suggest that trophoblast infection may be a mechanism of transplacental ebolavirus transmission. Published by Oxford University Press for the Infectious Diseases Society of America 2016. This work is written by (a) US Government employee(s) and is in the public domain in the US.

  5. Immunohistochemical Assessment of HER3 Expression in Odontogenic Cysts

    Science.gov (United States)

    Honarmand, Marieh; Saravani, Shirin; Kamyab, Nazanin; Jahantigh, Mehdi; Torabi Parizi, Molouk

    2015-01-01

    Background: It has been demonstrated that HER3 plays an important role in some human cancers and the HER3 expression is associated with worse survival in solid tumors. Objectives: This study was conducted to compare HER3 expression in epithelial lining of radicular cysts (RCs), dentigerous cysts (DCs) and odontogenic keratocysts (OKCs). Materials and Methods: This was a descriptive-analytical study, which assessed all 57 paraffin blocks of RCs, DCs and OKCs (21 RCs, 16 DCs, 20 OKC) from pathological archive of Dentistry College of Zahedan, Iran. The HER3 expression in cytoplasm and membrane was examined by immunohistochemical method. The data collected was analyzed using SPSS16 by ANOVA and Chi-square. P cysts was higher than that in inflammatory odontogenic cysts. The higher rate of HER3 expression in OKC may justify inherent growth potential, stimulation-independent proliferation capability, invasive growth and high recurrence rate of the cyst accepted today as a tumor. PMID:26734469

  6. Immunohistochemical study of Metallothionein in patients with temporal lobe epilepsy.

    Science.gov (United States)

    Juárez-Rebollar, Daniel; Alonso-Vanegas, Mario; Nava-Ruíz, Concepción; Buentello-García, Masao; Yescas-Gómez, Petra; Díaz-Ruíz, Araceli; Rios, Camilo; Méndez-Armenta, Marisela

    2017-05-01

    Epilepsy is characterized by spontaneous recurrent seizures and temporal lobe epilepsy (TLE) is the most common serious neurological example of acquired and frequent epilepsy. Oxidative stress is recognized as playing a contributing role in several neurological disorders, and most recently have been implicated in acquired epilepsies. The MTs occur in several brain regions and may serve as neuroprotective proteins against reactive oxygen species causing oxidative damage and stress. The main aim of this work was to describe the immunohistochemical localization of MT in the specimens derived from the patients affected by TLE. Histopathological examination showed NeuN, GFAP and MT immunopositive cells that were analyzed for determinate in hippocampal and parietal cortex samples. An increase in the reactive gliosis associated with increased MT expression was observed in patients with TLE. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Antigenic heterogeneity in granular cell ameloblastoma: an immunohistochemical study.

    Science.gov (United States)

    Ramya, Ramadoss; Sudheerkanth, Peddanna Kondamari; Eapen, Cherian; Alex, Varghese Kumaranthara; Rajkumar, Krishnan; Rajashree, Padmanaban

    2015-03-01

    Nature of granular cells in granular cell ameloblastoma (GCA) has always invoked considerable interest. The present study aims at antigenic characterization in five such cases with a panel of markers. Tissue specimens of five patients with GCA were fixed in buffered formalin and later embedded in paraffin wax. Blocks were sliced into 3 µ thick sections for immunohistochemical analysis using a panel of markers CD68, Bcl2, S100, p53, cytokeratin (AE1/AE3), vimentin and desmin. All five cases were strongly positive for cytokeratin and CD68. S100 was negative in three cases and showed a mild positivity in two cases. Bcl2, p53, vimentin and desmin were negative in all the five cases. This study presents a heterogenous nature of the granular cells; however, further validation is required with a larger sample size.

  8. The dark side of the immunohistochemical moon: industry.

    Science.gov (United States)

    Kalyuzhny, Alexander E

    2009-12-01

    Modern biological research is dependent on tools developed and provided by commercial suppliers, and antibodies for immunohistochemistry are among the most frequently used of these tools. Not all commercial antibodies perform as expected, however; this problem leads researchers to waste time and money when using antibodies that perform inadequately. Different commercial suppliers offer antibodies of varying degrees of quality and, in some cases, are unable to provide expert technical support for the immunohistochemical use of their antibodies. This article briefly describes the production of commercial antibodies from the manufacturer's perspective and presents some guidelines for choosing appropriate commercial antibodies for immunohistochemistry. Additionally, the article suggests steps to establish mutually beneficial relationships between commercial antibody suppliers and researchers who use them.

  9. Quantifying mast cells in bladder pain syndrome by immunohistochemical analysis

    DEFF Research Database (Denmark)

    Larsen, M.S.; Mortensen, S.; Nordling, J.

    2008-01-01

    OBJECTIVES To evaluate a simple method for counting mast cells, thought to have a role in the pathophysiology of bladder pain syndrome (BPS, formerly interstitial cystitis, a syndrome of pelvic pain perceived to be related to the urinary bladder and accompanied by other urinary symptoms, e. g....... frequency and nocturia), as > 28 mast cells/mm(2) is defined as mastocytosis and correlated with clinical outcome. PATIENTS AND METHODS The current enzymatic staining method (naphtolesterase) on 10 mu m sections for quantifying mast cells is complicated. In the present study, 61 patients had detrusor...... sections between, respectively. Mast cells were counted according to a well-defined procedure. RESULTS The old and the new methods, on 10 and 3 mu m sections, showed a good correlation between mast cell counts. When using tryptase staining and 3 mu m sections, the mast cell number correlated well...

  10. Immunohistochemical detection of C9orf72 protein in frontotemporal lobar degeneration and motor neurone disease: patterns of immunostaining and an evaluation of commercial antibodies.

    Science.gov (United States)

    Davidson, Yvonne S; Robinson, Andrew C; Rollinson, Sara; Pickering-Brown, Stuart; Xiao, Shangxi; Robertson, Janice; Mann, David M A

    2018-02-01

    We have employed as 'gold standards' two in-house, well-characterised and validated polyclonal antibodies, C9-L and C9-S, which detect the longer and shorter forms of C9orf72, and have compared seven other commercially available antibodies with these in order to evaluate the utility of the latter as credible tools for the demonstration of C9orf72. C9-L and C9-S antibodies immunostained cytoplasmic 'speckles', and the nuclear membrane, respectively, in cerebellar Purkinje cells of the cerebellum in patients with behavioural variant frontotemporal dementia (bvFTD) with amyotrophic lateral sclerosis (ALS), and in patients with ALS alone. Similar staining was seen in Purkinje cells in healthy control tissues and in other neurodegenerative disorders, and in pyramidal cells of CA4 and dentate gyrus of hippocampus. However, in the spinal cord there was little cytoplasmic staining with C9-L antibody. C9-S antibody immunostained the nuclear membrane of anterior horn cells in healthy neurons. In patients with bvFTD + ALS, or ALS alone, this C9-S nuclear staining was redistributed to the plasma membrane. In those patients with bvFTD + ALS or ALS bearing an expansion in C9orf72, none of the commercially available antibodies detected TDP-43 inclusions in anterior horn cells, nor were dipeptide repeat proteins demonstrated. Five of the commercial antibodies provided immunohistochemical staining patterns similar in morphological appearance to the in-house C9-L antibody, but distinct from C9-S antibody. However, only three showed sufficient specificity and intensity of staining for C9orf72 at acceptably low concentrations, to make them of practical value and sufficiently reliable for the detection of at least the longer form of C9orf72 protein.

  11. Immunohistochemical detection of intra-neuronal VZV proteins in snap-frozen human ganglia is confounded by antibodies directed against blood group A1-associated antigens.

    Science.gov (United States)

    Ouwendijk, Werner J D; Flowerdew, Sarah E; Wick, Desiree; Horn, Anja K E; Sinicina, Inga; Strupp, Michael; Osterhaus, Albert D M E; Verjans, Georges M G M; Hüfner, Katharina

    2012-06-01

    Varicella-zoster virus (VZV) causes chickenpox, establishes latency in trigeminal (TG) and dorsal root ganglia (DRG), and can lead to herpes zoster upon reactivation. The VZV proteome expressed during latency remains ill-defined, and previous studies have shown discordant data on the spectrum and expression pattern of VZV proteins and transcripts in latently infected human ganglia. Recently, Zerboni and colleagues have provided new insight into this discrepancy (Zerboni et al. in J Virol 86:578-583, 2012). They showed that VZV-specific ascites-derived monoclonal antibody (mAb) preparations contain endogenous antibodies directed against blood group A1 proteins, resulting in false-positive intra-neuronal VZV staining in formalin-fixed human DRG. The aim of the present study was to confirm and extend this phenomenon to snap-frozen TG (n=30) and DRG (n=9) specimens of blood group genotyped donors (n=30). The number of immunohistochemically stained neurons was higher with mAb directed to immediate early protein 62 (IE62) compared with IE63. The IE63 mAb-positive neurons always co-stained for IE62 but not vice versa. The mAb staining was confined to distinct large intra-neuronal vacuoles and restricted to A1(POS) donors. Anti-VZV mAb staining in neurons, but not in VZV-infected cell monolayers, was obliterated after mAb adsorption against blood group A1 erythrocytes. The data presented demonstrate that neuronal VZV protein expression detected by ascites-derived mAb in snap-frozen TG and DRG of blood group A1(POS) donors can be misinterpreted due to the presence of endogenous antibodies directed against blood group A1-associated antigens present in ascites-derived VZV-specific mAb preparations.