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Sample records for histopathology tissue samples

  1. A low dimensional entropy-based descriptor of several tissues in skin cancer histopathology samples

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    Álvarez, Pablo; Corredor, Germán.; García-Arteaga, Juan D.; Romero, Eduardo

    2015-12-01

    The use of low-level visual features to assign high level labels in datasets of histopathology images is a possible solution to the problems derived from manual labeling by experts. However, in many cases, the visual cues are not enough. In this article we propose the use of features derived exclusively from the spatial distribution of the cell nuclei. These features are calculated using the weight of k-nn graphs constructed from the distances between cells. Results show that there are k values with enhanced discriminatory power, especially when comparing cancerous and non-cancerous tissue.

  2. Critical steps in tissue processing in histopathology.

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    Comanescu, Maria; Annaratone, Laura; D'Armento, Giuseppe; Cardos, Georgeta; Sapino, Anna; Bussolati, Gianni

    2012-04-01

    Histopathological diagnosis using Formalin-Fixed Paraffin Embedded (FFPE) tissues is essential for the prognostic and therapeutic management of cancer patients. Pathologists are being confronted with increasing demands, from both clinicians and patients, to provide immunophenotypic and gene expression data from FFPE tissues to allow the planning of personalized therapeutic regimens. Recent improvements in the protocols for pre-analysis processing of pathological tissues aim to better preserve cellular details and to conserve antigens and nucleic acid sequences. These developments have been recently patented. The international protocol for the transporting of surgical specimens from the surgical theatre to the pathology department is to immerse the specimen in formalin. The alternative method of sealing the specimens into bags under a vacuum and then cooling is a well-accepted and environmentally safe procedure that overcomes the many drawbacks linked to transfer in formalin. Importantly, RNA is notoriously poorly preserved in FFPE tissue. Due to this, successful procedures for the extraction of genetic information from archival tissues have been the object of several studies and patents. Novel molecular approaches for RT-qPCR and gene array analysis on FFPE tissues are presented here. Moreover, a major advance is reported in this study, the observation that tissue fixation in cold conditions allows a much better preservation of nucleic acid sequences.

  3. Histopathology of mucosa-associated lymphoid tissue

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    Kuper, C.F.

    2006-01-01

    Mucosa-associated lymphoid tissue (MALT) is a generalized term incorporating a disseminated collection of lymphoid tissues in multiple sites throughout the body. MALT sites that have been/are primarily studied include bronchus-associated lymphoid tissue (BALT), gut-associated lymphoid tissue (GALT),

  4. DNA qualification workflow for next generation sequencing of histopathological samples.

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    Michele Simbolo

    Full Text Available Histopathological samples are a treasure-trove of DNA for clinical research. However, the quality of DNA can vary depending on the source or extraction method applied. Thus a standardized and cost-effective workflow for the qualification of DNA preparations is essential to guarantee interlaboratory reproducible results. The qualification process consists of the quantification of double strand DNA (dsDNA and the assessment of its suitability for downstream applications, such as high-throughput next-generation sequencing. We tested the two most frequently used instrumentations to define their role in this process: NanoDrop, based on UV spectroscopy, and Qubit 2.0, which uses fluorochromes specifically binding dsDNA. Quantitative PCR (qPCR was used as the reference technique as it simultaneously assesses DNA concentration and suitability for PCR amplification. We used 17 genomic DNAs from 6 fresh-frozen (FF tissues, 6 formalin-fixed paraffin-embedded (FFPE tissues, 3 cell lines, and 2 commercial preparations. Intra- and inter-operator variability was negligible, and intra-methodology variability was minimal, while consistent inter-methodology divergences were observed. In fact, NanoDrop measured DNA concentrations higher than Qubit and its consistency with dsDNA quantification by qPCR was limited to high molecular weight DNA from FF samples and cell lines, where total DNA and dsDNA quantity virtually coincide. In partially degraded DNA from FFPE samples, only Qubit proved highly reproducible and consistent with qPCR measurements. Multiplex PCR amplifying 191 regions of 46 cancer-related genes was designated the downstream application, using 40 ng dsDNA from FFPE samples calculated by Qubit. All but one sample produced amplicon libraries suitable for next-generation sequencing. NanoDrop UV-spectrum verified contamination of the unsuccessful sample. In conclusion, as qPCR has high costs and is labor intensive, an alternative effective standard

  5. DNA qualification workflow for next generation sequencing of histopathological samples.

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    Simbolo, Michele; Gottardi, Marisa; Corbo, Vincenzo; Fassan, Matteo; Mafficini, Andrea; Malpeli, Giorgio; Lawlor, Rita T; Scarpa, Aldo

    2013-01-01

    Histopathological samples are a treasure-trove of DNA for clinical research. However, the quality of DNA can vary depending on the source or extraction method applied. Thus a standardized and cost-effective workflow for the qualification of DNA preparations is essential to guarantee interlaboratory reproducible results. The qualification process consists of the quantification of double strand DNA (dsDNA) and the assessment of its suitability for downstream applications, such as high-throughput next-generation sequencing. We tested the two most frequently used instrumentations to define their role in this process: NanoDrop, based on UV spectroscopy, and Qubit 2.0, which uses fluorochromes specifically binding dsDNA. Quantitative PCR (qPCR) was used as the reference technique as it simultaneously assesses DNA concentration and suitability for PCR amplification. We used 17 genomic DNAs from 6 fresh-frozen (FF) tissues, 6 formalin-fixed paraffin-embedded (FFPE) tissues, 3 cell lines, and 2 commercial preparations. Intra- and inter-operator variability was negligible, and intra-methodology variability was minimal, while consistent inter-methodology divergences were observed. In fact, NanoDrop measured DNA concentrations higher than Qubit and its consistency with dsDNA quantification by qPCR was limited to high molecular weight DNA from FF samples and cell lines, where total DNA and dsDNA quantity virtually coincide. In partially degraded DNA from FFPE samples, only Qubit proved highly reproducible and consistent with qPCR measurements. Multiplex PCR amplifying 191 regions of 46 cancer-related genes was designated the downstream application, using 40 ng dsDNA from FFPE samples calculated by Qubit. All but one sample produced amplicon libraries suitable for next-generation sequencing. NanoDrop UV-spectrum verified contamination of the unsuccessful sample. In conclusion, as qPCR has high costs and is labor intensive, an alternative effective standard workflow for

  6. Ensemble Learning of Tissue Components for Prostate Histopathology Image Grading

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    Dheeb Albashish

    2016-12-01

    Full Text Available Ensemble learning is an effective machine learning approach to improve the prediction performance by fusing several single classifier models. In computer-aided diagnosis system (CAD, machine learning has become one of the dominant solutions for tissue images diagnosis and grading. One problem in a single classifier model for multi-components of the tissue images combination to construct dense feature vectors is the overfitting. In this paper, an ensemble learning for multi-component tissue images classification approach is proposed. The prostate cancer Hematoxylin and Eosin (H&E histopathology images from HUKM were used to test the proposed ensemble approach for diagnosing and Gleason grading. The experiments results of several prostate classification tasks, namely, benign vs. Grade 3, benign vs.Grade4, and Grade 3vs.Grade 4 show that the proposed ensemble significantly outperforms the previous typical CAD and the naïve approach that combines the texture features of all tissue component directly in dense feature vectors for a classifier.

  7. The mandate for a proper preservation in histopathological tissues.

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    Comănescu, Maria; Arsene, D; Ardeleanu, Carmen; Bussolati, G

    2012-01-01

    A sequence of technically reproducible procedures is mandatory to guarantee a proper preservation of tissues and to build up the basis for sound diagnoses. However, while the goal of these procedures was, until recently, to assure only structural (histological and cytological) preservation, an appropriate preservation of antigenic properties and of nucleic acid integrity is now additionally requested, in order to permit pathologists to provide the biological information necessary for the adoption of personalized therapies. The present review analyses the sequence of technical steps open to critical variations. Passages such as dehydration, paraffin embedding, sectioning and staining are relatively well standardized and allow adoption of dedicated (automatic) apparatuses, while other pre-analytical steps, i.e. time and modalities of transfer of surgical specimens from the surgical theatre to the pathology laboratory (s.c. "ischemia time") and the type and length of fixation are not standardized and are a potential cause of discrepancies in diagnostic results. Our group is involved in European-funded projects tackling these problems with the concrete objective of implementing a model of effective tumors investigations by high performance genetic and molecular methodologies. The problem of the discrepant quality level of histopathological and cytological preparations involved five European countries and exploiting the potential of "virtual slide technology". Concrete issues, techniques and pitfalls, as well as proposed guidelines for processing the tissues are shown in this presentation.

  8. Histopathological changes in supraspinous ligaments, ligamentum flava and paraspinal muscle tissues of patients with ankylosing spondylitis.

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    Zhang, Ying; Xu, Hongfei; Hu, Xu; Zhang, Chao; Chu, Tongwei; Zhou, Yue

    2016-04-01

    To examine the histopathological changes in spinal tissues of ankylosing spondylitis (AS) patients. Tissue samples from 10 AS patients and 10 control subjects were obtained. Hematoxylin and eosin, picrosirius, Masson and van Gieson stainings were utilized to determine the pathological changes in tissues. Ultrastructural alterations were examined by electronic microscopy. Proteoglycan levels were assessed by enzyme-linked immunosorbent assays (ELISA). Matrix metalloproteinase-3 (MMP-3), transforming growth factor-β1 (TGF-β1) and tumor necrosis factor-α (TNF-α) levels were evaluated by immunohistochemistry. Our results demonstrate that the density of collagen fibrils was reduced in the supraspinous ligaments of AS tissue and fibrils were loosely and irregularly organized as compared to a regular distribution of collagen fibrils in controls. In ligamentum flava from AS patients, activated fibroblasts with enlarged nuclei were detected, while the number of elastic fibers was greatly decreased. Paraspinal muscle tissues of AS patients exhibited increased collagen fibril accumulation and atrophy. Significantly decreased proteoglycan and elevated MMP-3 levels were found in supraspinous ligament samples from AS patients (P muscle tissues of AS patients were increased (P muscle between control and AS patients (P > 0.05). Our findings reveal histopathological changes that occur in certain spinal tissues of AS patients and suggest that increased levels of MMP-3 and TGF-β1 may contribute to the pathogenesis of AS. © 2014 The Authors. International Journal of Rheumatic Diseases published by Asia Pacific League of Associations for Rheumatology and Wiley Publishing Asia Pty Ltd.

  9. Revisiting peri-implant soft tissuehistopathological study of the peri-implant soft tissue

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    Silva, Eduarda; Félix, Sérgio; Rodriguez-Archilla, Alberto; Oliveira, Pedro; Martins dos Santos, José

    2014-01-01

    Peri-implant soft tissues are essential for osseointegration. The peri-implant mucosa may lack vascular supply, and histological observation, even without plaque, shows the presence of inflammatory cells. The objectives of this study were to assess the histopathological changes of the epithelium and connective tissue around the implant. Twenty patients of both genders were studied. Twelve weeks after implant placement, fragments of peri-implant gingival sulcus were harvested and processed for light microscopy. Group I (10): without clinical inflammatory signs (control); Group II (10): with clinical inflammatory signs. Histopathological parameters were analyzed and classified in 3 grades: mild, moderate or severe (grade 1, 2 or 3). Control group showed only slight changes, grade 1. In group II we found edema with moderate to severe cellular and nuclear changes. There are more women than men with all grades of inflammation. All patients with moderate edema are male and all patients with severe edema are female. A significant association (p=0.007) exists between these two variables. Significant differences were found when comparing the degree of inflammation with nuclear alterations (p=0.001) and the same results when comparing the degree of edema and nuclear changes (pimplant mucosa. In clinics the predisposition of female patients to greater degree of edema and inflammation should be accounted for. PMID:24551281

  10. [Comments on histopathology examinations in forensic medicine -- the importance, indications and basic principles of preserving the tissues].

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    Jankowski, Zbigniew; Pieśniak, Dorota

    2007-01-01

    Histopathology examinations and their usefulness in postmortem diagnosis and medico-legal expertise have always been regarded extremely important by the Faculty of the Chair and Department of Forensic Medicine at the Medical University of Gdansk. The authors postulate acceptance of indication criteria for histopathology examination in forensic autopsies and propose rules for collecting tissue samples from internal organs and the placenta. They also remind the readers of the principles of proceeding with the preserved material. The present paper may be helpful for doctors other than forensic pathologists, who perform autopsies in institutions other than university departments of forensic medicine.

  11. A Histopathological Study of Pulmonary Hypertension in Connective Tissue Disease

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    Nobuhito Sasaki

    2011-01-01

    Full Text Available Connective tissue diseases (CTD, such as systemic sclerosis (SSc, systemic lupus erythematosus (SLE, and mixed connective tissue disease (MCTD, develop pulmonary hypertension (PH. Generally all PH cases associated with any CTD are classified into the same PH group. However, histological examination shows both common and specific lesions for each disease. In patients with SLE, fibrosis is generally rare and mild. The findings of PH in SLE are similar to those in primary pulmonary hypertension. Many cases of SSc are accompanied by fibrosis. MCTD is rather close to SSc. Arterial and arteriolar lesions of MCTD are characterized by fibrous intimal thickening. In this review, we describe the pathological features of PH associated with each CTD.

  12. [Parasitological, immunohistochemical and histopathological study for Leishmania chagasi detection in splenic tissues of dogs with visceral leishmaniasis].

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    Tasca, Karen Ingrid; Buzetti, Wilma Aparecida Starke; Tenorio, Michely da Silva; Paulan, Silvana de Cássia; Lima, Flávia Luna; de Queiroz, Nina Mari Gual Pimenta; Machado, Rosângela Zacarias; Oliveira, Tricia Maria Ferreira de Souza; Neves, Maria Francisca; de Noronha, Antonio Carlos Faconti; de Assis, Juliana

    2009-01-01

    The purpose of this work was a Canine Visceral Leishmaniasis--CVL study by parasitological direct examination of Leishmania (L.) chagasi (imprinting and histological), immunohistochemical test and histopathological analysis using spleen tissues from 34 dogs euthanized by the Zoonotic Disease Control Centre from Ilha Solteira, SP, Brazil. According to the clinical signs, the dogs were divided in three groups: asymptomatics (8 dogs), oligosymptomatics (17 dogs) and symptomatics (9 dogs). After the accomplishment of all diagnostic tests, 22 dogs were considered positives (64.7%) and 12 (35.3%) were negatives to CVL. From these positive dogs, 1/22 (4.5%) was asymptomatic, 12/22(54.5%) were oligosymptomatics and 8/22 (40.1%) were symptomatics. The histopathological study in spleen tissues from positive, especially symptomatic dogs, showed a diffuse chronic inflammation with thickness of capsular and trabecular regions and there was extensive morphologic alteration of the red and white pulp by the presence of abundant macrophages full with amastigotes, the granulomatous inflammatory reaction and haemorrhagic areas. The data of this work from histopathologic examination and direct microscopic visualization of L. (L.) chagasi showed that the spleen was an useful organ to collect sample tissues for CVL diagnosis. The immunostaining detected the highest number of positive dogs and were considered an important and conclusive method to be used in addition to parasitological methods for CVL, particularly in asymptomatic or oligosymptomatic dogs.

  13. Histopathological changes in the gill and liver tissues of freshwater fish, Cirrhinus mrigala exposed to dichlorvos

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    Babu Velmurugan

    2009-10-01

    Full Text Available The histopathological effects of dichlorvos, an organophosphate pesticide, on the gill and liver tissues in Cirrhinus mrigala were determined by light microscopy. The fishes were exposed to sublethal concentrations (0.91 and 1.82 ppm of dichlorvos for 10 days with parallel untreated control. No histopathological effects were observed in control group. Hyperplasia, desquamation, and necrosis of epithelial, epithelial lifting, oedema, lamellar fusion, collapsed secondary lamellae, curling of secondary lamellae and aneurism in the secondary lamellae were observed in gill tissues exposed to dichlorvos. Hepatic lesions in the liver tissues of fishes exposed to dichlorvos were characterized by cloudy swelling of hepatocytes, congestion, vacuolar degeneration, karyolysis, karyohexis, dilation of sinusoids and nuclear hypertrophy.

  14. Extensive HIV-1 intra-host recombination is common in tissues with abnormal histopathology.

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    Susanna L Lamers

    Full Text Available There is evidence that immune-activated macrophages infected with the Human Immunodeficiency Virus (HIV are associated with tissue damage and serve as a long-lived viral reservoir during therapy. In this study, we analyzed 780 HIV genetic sequences generated from 53 tissues displaying normal and abnormal histopathology. We found up to 50% of the sequences from abnormal lymphoid and macrophage rich non-lymphoid tissues were intra-host viral recombinants. The presence of extensive recombination, especially in non-lymphoid tissues, implies that HIV-1 infected macrophages may significantly contribute to the generation of elusive viral genotypes in vivo. Because recombination has been implicated in immune evasion, the acquisition of drug-resistance mutations, and alterations of viral co-receptor usage, any attempt towards the successful eradication of HIV-1 requires therapeutic approaches targeting tissue macrophages.

  15. Histopathologic changes in soft tissue associated with radiographically normal impacted third molars

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    Kotrashetti Vijayalakshmi

    2010-01-01

    Full Text Available Background: The incidence of impacted or embedded third molars accounts for approximately 98%. Since 1948, there are studies reporting pathological changes in an asymptomatic dental follicle. Controversy still exists for removal of asmptomatic impacted teeth. Hence, this study was performed to histologically evaluate soft tissue pathosis in the pericoronal tissues of impacted third molars with pericoronal radiolucency measuring up to 2.5 mm on orthopantomographs. Materials and Methods: Forty-one asymptomatic impacted third molars with follicular space of up to 2.5 mm on radiographs were included. The disimpacted teeth and the follicular tissues were obtained for histological examination. Results: Age of the patients ranged from 14 to 25 years. Of 41 tissues evaluated, histopathological reports of 18 follicles were suggestive of dentigerous cyst, two follicles showed odontogenic keratocyst, one follicle each of calcifying epithelial odontogenic cyst, ameloblastoma-like proliferation, odontogenic myxoma and odontogenic fibroma. Conclusion: This study showed 58.5% of asymptomatic cases with definite pathological changes. Hence, thorough clinical and radiographic examination should be carried out for all impacted third molars and the dental follicular tissue should be submitted for histopathological evaluation.

  16. Two-tier tissue decomposition for histopathological image representation and classification.

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    Gultekin, Tunc; Koyuncu, Can Fahrettin; Sokmensuer, Cenk; Gunduz-Demir, Cigdem

    2015-01-01

    In digital pathology, devising effective image representations is crucial to design robust automated diagnosis systems. To this end, many studies have proposed to develop object-based representations, instead of directly using image pixels, since a histopathological image may contain a considerable amount of noise typically at the pixel-level. These previous studies mostly employ color information to define their objects, which approximately represent histological tissue components in an image, and then use the spatial distribution of these objects for image representation and classification. Thus, object definition has a direct effect on the way of representing the image, which in turn affects classification accuracies. In this paper, our aim is to design a classification system for histopathological images. Towards this end, we present a new model for effective representation of these images that will be used by the classification system. The contributions of this model are twofold. First, it introduces a new two-tier tissue decomposition method for defining a set of multityped objects in an image. Different than the previous studies, these objects are defined combining texture, shape, and size information and they may correspond to individual histological tissue components as well as local tissue subregions of different characteristics. As its second contribution, it defines a new metric, which we call dominant blob scale, to characterize the shape and size of an object with a single scalar value. Our experiments on colon tissue images reveal that this new object definition and characterization provides distinguishing representation of normal and cancerous histopathological images, which is effective to obtain more accurate classification results compared to its counterparts.

  17. 7-tesla MR imaging of non-melanoma skin cancer samples: correlation with histopathology.

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    Aubry, Sébastien; Leclerc, Olga; Tremblay, Luc; Rizcallah, Edmond; Croteau, Frédéric; Orfali, Charles; Lepage, Martin

    2012-11-01

    The aims of this study were to compare in vitro magnetic resonance imaging (MRI) characteristics of keratinocytic skin cancer assessed by a 7-tesla (T) MRI with histopathology, and to describe MRI features of skin tumors. This prospective study included 30 skin tumors treated by surgery. MR images of skin samples were acquired on a 7-T MR scanner using a fast spin-echo T(2)-weighted and an isotropic 3D gradient-echo T(1)-weighted sequence. Length, width, Breslow index and margins of the lesions were measured. The presence or absence of the following was noted: healthy margins, ulceration of the dermis, in situ lesions, superficial and deep dermis involvement, subcutaneous involvement, superficial and intratumoral keratin. MR results were compared to histopathology. Interclass correlation coefficient (ICC) was very good for the evaluation of the width (ICC = 0.86) and Breslow index (ICC = 0.87). The ICC was good for the evaluation of the margins (ICC = 0.70) but for length, ICC was lower (ICC = 0.67). Mean bias between MRI and histopathology was inferior to 1 mm for width, Breslow index and margin. In vitro 7-T MRI of keratinocytic skin cancer allows delineation of lesions with good correlation with histopathology. After in vivo confirmation it could have a diagnostic role regarding the delineation of surgical margins but its actual limitations prevent its practical adoption at this time. © 2011 John Wiley & Sons A/S.

  18. Histopathology effects of nickel nanoparticles on lungs, liver, and spleen tissues in male mice

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    Ajdari, Marziyeh; Ziaee Ghahnavieh, Marziyeh

    2014-09-01

    Because of the classification of the nickel compounds as carcinogenic substances, there is a need for in vivo tests to nickel nanoparticles (NiNPs) for observing their effects on health experimentally. Spherical NiNPs with 10 nm in diameter and 75 ppm concentration were applied for investigating their toxicities within male albino mice as an in vivo model. We randomly made sham group, control group, and 75 ppm group (with five animals in each group). Then, the nanoparticles were injected into mice intraperitonealy for 7 days and after that their lungs, liver, and spleen were removed for histopathological observations. At the end of the test, section microscopic observations of liver, spleen, and lung in sham and control groups showed normal tissues but these tissues underwent significant abnormal effects in 75 ppm group. NiNPs can cause undesirable effects in lungs, liver, and spleen tissues with same condition of this study.

  19. Is tissue an issue? Current practice and opinion in Western Australia for routine histopathology on products of conception.

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    Yap, Shui-Jean; Watts, Jared C; Faithfull, Tiffany J; Wong, Sabrina Z; Wylde, Kate L; McGurgan, Paul M

    2014-10-01

    An anonymous questionnaire-based survey was used to determine current practices and opinions of senior health professionals working in Western Australian (WA) hospitals performing gynaecological procedures, regarding the routine use of histopathology for products of conception (POC) obtained either from the surgical management of miscarriage or termination of pregnancy. Sixty-one senior health professionals completed the survey. Tissue histopathology on POC was routinely requested for miscarriage and termination of pregnancy (TOP) by 87 and 59% of respondents, respectively. Respondents listed the main reasons for requesting routine histopathology as avoidance of misdiagnosis, medico-legal and quality assurance. There were inconsistent practices among WA health professionals regarding sending POC for histopathology; 63% of gynaecology head of departments recommend the introduction of state or national guidelines for the use of histopathology in the surgical management of miscarriages or terminations of pregnancy.

  20. Dielectric characterisation of human tissue samples

    NARCIS (Netherlands)

    Rossum, W.L. van; Nennie, F.; Deiana, D.; Veen, A.J. van der; Monni, S.

    2014-01-01

    The electrical properties of tissues samples are required for investigation and simulation purposes in biomedical applications of EM sensors. While available open literature mostly deals with ex-vivo characterization of isolated tissues, knowledge on dielectric properties of these tissues in their o

  1. Eliminating tissue-fold artifacts in histopathological whole-slide images for improved image-based prediction of cancer grade

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    Sonal Kothari

    2013-01-01

    Full Text Available Background: Analysis of tissue biopsy whole-slide images (WSIs depends on effective detection and elimination of image artifacts. We present a novel method to detect tissue-fold artifacts in histopathological WSIs. We also study the effect of tissue folds on image features and prediction models. Materials and Methods: We use WSIs of samples from two cancer endpoints - kidney clear cell carcinoma (KiCa and ovarian serous adenocarcinoma (OvCa - publicly available from The Cancer Genome Atlas. We detect tissue folds in low-resolution WSIs using color properties and two adaptive connectivity-based thresholds. We optimize and validate our tissue-fold detection method using 105 manually annotated WSIs from both cancer endpoints. In addition to detecting tissue folds, we extract 461 image features from the high-resolution WSIs for all samples. We use the rank-sum test to find image features that are statistically different among features extracted from the same set of WSIs with and without folds. We then use features that are affected by tissue folds to develop models for predicting cancer grades. Results: When compared to the ground truth, our method detects tissue folds in KiCa with 0.50 adjusted Rand index (ARI, 0.77 average true rate (ATR, 0.55 true positive rate (TPR, and 0.98 true negative rate (TNR; and in OvCa with 0.40 ARI, 0.73 ATR, 0.47 TPR, and 0.98 TNR. Compared to two other methods, our method is more accurate in terms of ARI and ATR. We found that 53 and 30 image features were significantly affected by the presence of tissue-fold artifacts (detected using our method in OvCa and KiCa, respectively. After eliminating tissue folds, the performance of cancer-grade prediction models improved by 5% and 1% in OvCa and KiCa, respectively. Conclusion: The proposed connectivity-based method is more effective in detecting tissue folds compared to other methods. Reducing tissue-fold artifacts will increase the performance of cancer-grade prediction

  2. Preparation of tissue samples for X-ray fluorescence microscopy

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    Chwiej, Joanna [Faculty of Physics and Applied Computer Science, AGH-University of Science and Technology, Al. Mickiewicza 30, 30-059 Cracow (Poland)]. E-mail: jchwiej@novell.ftj.agh.edu.pl; Szczerbowska-Boruchowska, Magdalena [Faculty of Physics and Applied Computer Science, AGH-University of Science and Technology, Al. Mickiewicza 30, 30-059 Cracow (Poland); Lankosz, Marek [Faculty of Physics and Applied Computer Science, AGH-University of Science and Technology, Al. Mickiewicza 30, 30-059 Cracow (Poland); Wojcik, Slawomir [Faculty of Physics and Applied Computer Science, AGH-University of Science and Technology, Al. Mickiewicza 30, 30-059 Cracow (Poland); Falkenberg, Gerald [Hamburger Synchrotronstrahlungslabor at Deutsches Elektronen-Synchrotron, Notkestr. 85, Hamburg (Germany); Stegowski, Zdzislaw [Faculty of Physics and Applied Computer Science, AGH-University of Science and Technology, Al. Mickiewicza 30, 30-059 Cracow (Poland); Setkowicz, Zuzanna [Department of Neuroanatomy, Institute of Zoology, Jagiellonian University, Ingardena 6, 30-060 Cracow (Poland)

    2005-12-15

    As is well-known, trace elements, especially metals, play an important role in the pathogenesis of many disorders. The topographic and quantitative elemental analysis of pathologically changed tissues may shed some new light on processes leading to the degeneration of cells in the case of selected diseases. An ideal and powerful tool for such purpose is the Synchrotron Microbeam X-ray Fluorescence technique. It enables the carrying out of investigations of the elemental composition of tissues even at the single cell level. The tissue samples for histopathological investigations are routinely fixed and embedded in paraffin. The authors try to verify the usefulness of such prepared tissue sections for elemental analysis with the use of X-ray fluorescence microscopy. Studies were performed on rat brain samples. Changes in elemental composition caused by fixation in formalin or paraformaldehyde and embedding in paraffin were examined. Measurements were carried out at the bending magnet beamline L of the Hamburger Synchrotronstrahlungslabor HASYLAB in Hamburg. The decrease in mass per unit area of K, Br and the increase in P, S, Fe, Cu and Zn in the tissue were observed as a result of the fixation. For the samples embedded in paraffin, a lower level of most elements was observed. Additionally, for these samples, changes in the composition of some elements were not uniform for different analyzed areas of rat brain.

  3. A handcrafted tissue microarray for a matrix arrangement of tissue samples.

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    Sampaio, João P A; Cavalcante, José R; Furtado, Francisco N N; Lima-Júnior, Roberto C P; Ribeiro, Ronaldo A; Almeida, Paulo R C

    2014-01-01

    Tissue microarray (TMA) was first designed to enable more efficient immunohistochemical screening of antibodies and tissues. However, due to the high cost of commercial TMA builder instrument, such method is not affordable for many pathology laboratories. Then, methodological adaptations have been proposed in order to reduce TMA-associated cost. A manual leather puncher with an inner diameter of 2mm was used to collect a tissue sample from the donor paraffin block. The conventional TMA method was adopted as a control group. Empty paraffin recipient blocks were prepared and a standard 2-mm crochet needle was used to create 24 equidistant holes in the recipient block. Tissue cores obtained from the donor blocks were transferred to the holes in the recipient blocks and routine histopathological techniques were then performed. In this study we proposed a new approach to produce TMA recipient blocks as an alternative to the conventional TMA. Copyright © 2014 Elsevier Inc. All rights reserved.

  4. Molecular and Histopathological Changes in Mouse Intestinal Tissue After Proton Exposure

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    Purgason, A.; Zhang, Y.; Wu, H.

    2010-01-01

    Radiation in space, including types from solar particle events (SPE's), poses serious health risks to astronauts and is especially dangerous for long duration missions. Protons are the most abundant particles in deep space and to date there is little known about the details of the negative consequences crew members will face upon exposure to them. This ongoing project involves a mouse model subjected to several minutes of proton radiation at an energy of 250 MeV and doses of 0 Gy, 0.1 Gy, 1 Gy, and 2 Gy. The gastrointestinal tract of each animal was dissected four hours post-irradiation and the small intestine was isolated and flash-frozen. Three specimens per dose were studied. Tissue was homogenized and RNA was isolated in order for cDNA synthesis and real-time PCR to be performed. Gene expression changes are currently being analyzed specific to mouse apoptosis. Immunohistochemistry will be used to confirm any significant changes found in the analyses. Immunohistochemistry is also being used to observe gamma H2AX staining to learn of any DNA damage that occurred as a result of proton exposure. We expect to see increased DNA damage due to proton exposure. Finally, histopathologic observation of the tissue will be completed using standard H&E staining methods to screen for morphologic changes. Increased apoptosis is expected to be seen in the tissues which is typical of radiation damage. Observations will be confirmed by a pathologist.

  5. [General principles of handling tissues and organs intended for examination in histopathology - pathologists requirements for surgeons].

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    Trnková, M

    2014-03-01

    Histopathology has continued to develop into a complex discipline of laboratory medicine in the last 30 years. Implementation of new techniques such as immunohistochemistry, in situ hybridization, molecular pathology and gene profiling yields a large amount of information which is used not only to establish diagnosis, but also for prediction and prognosis. The basic conditions for precision and correctness of this information which directly influence the choice of therapy and the outcome for the patient, and eventually the health and the life of the patient, are obtaining a suitable specimen and the best laboratory processing. The first step in this procedure is collection of samples, fixation and submission to the pathology laboratory - the pre-analytical phase. Knowledge of the principles of this phase and their implementation in daily practice in surgery is of main importance for the quality and quantity of information obtained. We introduce the basic rules for proper handling of different specimens depending on the use of conventional and "new" histopathology methods.

  6. Graph-based pancreatic islet segmentation for early type 2 diabetes mellitus on histopathological tissue.

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    Floros, Xenofon; Fuchs, Thomas J; Rechsteiner, Markus P; Spinas, Giatgen; Moch, Holger; Buhmann, Joachim M

    2009-01-01

    It is estimated that in 2010 more than 220 million people will be affected by type 2 diabetes mellitus (T2DM). Early evidence indicates that specific markers for alpha and beta cells in pancreatic islets of Langerhans can be used for early T2DM diagnosis. Currently, the analysis of such histological tissues is manually performed by trained pathologists using a light microscope. To objectify classification results and to reduce the processing time of histological tissues, an automated computational pathology framework for segmentation of pancreatic islets from histopathological fluorescence images is proposed. Due to high variability in the staining intensities for alpha and beta cells, classical medical imaging approaches fail in this scenario. The main contribution of this paper consists of a novel graph-based segmentation approach based on cell nuclei detection with randomized tree ensembles. The algorithm is trained via a cross validation scheme on a ground truth set of islet images manually segmented by 4 expert pathologists. Test errors obtained from the cross validation procedure demonstrate that the graph-based computational pathology analysis proposed is performing competitively to the expert pathologists while outperforming a baseline morphological approach.

  7. Histopathologic effects of hypofractionated robotic radiation therapy on malignant and benign prostate tissue.

    Science.gov (United States)

    Suy, Simeng; Oermann, Eric; Hanscom, Heather; Lei, Sue; Vahdat, Saloomeh; Yu, Xia; Park, Hyeon U; Chen, Viola; Collins, Brian T; McGeagh, Kevin; Dawson, Nancy; Jha, Reena; Azumi, Norio; Dritschilo, Anatoly; Lynch, John; Collins, Sean P

    2010-12-01

    We describe the first histopathologic analysis of prostatic tissue following hypofractionated robotic radiation therapy. A 66 year-old man presented with stage II, low risk adenocarcinoma of the prostate and underwent elective conformal hypofractionated radiation therapy. His pretreatment evaluation revealed T1c adenocarcinoma, Gleason's grade 3 + 3 = 6 and a prostate specific antigen (PSA) level of 4.87 ng/ml. Hypofractionated radiation therapy (37.5 Gy in five daily fractions of 7.5 Gy) was completed on an Internal Review Board approved protocol. One year later, he developed progressive urinary retention. Transurethral prostatic resection was performed to alleviate obstructive symptoms. Bilobar hypertrophy was observed without evidence of stricture. Histolopathologic analyses of resected prostate tissues revealed changes consistent with radiation treatment, including cellular changes, inflammation, glandular atrophy and hyperplasia. There was no evidence of residual cancer, fibrosis or necrosis. The patient's postoperative course was uneventful with post-treatment PSA of 0.5 ng/ml and residual grade 1 stress incontinence.

  8. A survey of spontaneous occurrence of ochratoxin A residues in chicken tissues and concurrence with histopathological changes in liver and kidneys

    Science.gov (United States)

    Milicevic, Dragan; Jovanovic, Milijan; Matekalosverak, Vesna; Radicevic, Tatjana; Petrovic, Milan M.; Lilic, Slobodan

    2011-01-01

    Toxicological and histopathological investigations of tissues of commercially slaughtered chickens were carried out to provide a preliminary evaluation of the incidence of occurrence of ochratoxin A (OTA) in chicken sold in Serbian retail market. In addition, the etiology of nephropathies of these chickens was elucidated. The majority of these tissue samples were not found to contain measurable amounts of OTA. Moreover, the OTA levels found in analyzed tissues were generally low and there was no positive correlation between the presence of OTA and the frequency of histopathological changes. Histopathological changes such as degenerative changes in the kidneys and liver differed from the classical description of the mycotoxic nephropathy, indicating that the chicken nephropathy observed in Serbia may have a multitoxic etiology with possible synergistic effect between microorganisms and natural toxins, usually present in low concentrations. The low OTA results also suggested that chicken meat available in the retail market in Serbia are unlikely to pose any significant adverse health risk to the consumers with respect to OTA toxicity.

  9. Histopathological Analysis of PEEK Wear Particle Effects on the Synovial Tissue of Patients

    Directory of Open Access Journals (Sweden)

    A. C. Paulus

    2016-01-01

    Full Text Available Introduction. Increasing interest developed in the use of carbon-fiber-reinforced-poly-ether-ether-ketones (CFR-PEEK as an alternative bearing material in knee arthroplasty. The effects of CFR-PEEK wear in in vitro and animal studies are controversially discussed, as there are no data available concerning human tissue. The aim of this study was to analyze human tissue containing CFR-PEEK as well as UHMWPE wear debris. The authors hypothesized no difference between the used biomaterials. Methods and Materials. In 10 patients during knee revision surgery of a rotating-hinge-knee-implant-design, synovial tissue samples were achieved (tibial inserts: UHMWPE; bushings and flanges: CFR-PEEK. One additional patient received revision surgery without any PEEK components as a control. The tissue was paraffin-embedded, sliced into 2 μm thick sections, and stained with hematoxylin and eosin in a standard process. A modified panoptical staining was also done. Results. A “wear-type” reaction was seen in the testing and the control group. In all samples, the UHMWPE particles were scattered in the tissue or incorporated in giant cells. CFR-PEEK particles were seen as conglomerates and only could be found next to vessels. CFR-PEEK particles showed no giant-cell reactions. In conclusion, the hypothesis has to be rejected. UHMWPE and PEEK showed a different scatter-behavior in human synovial tissue.

  10. Histopathological Analysis of PEEK Wear Particle Effects on the Synovial Tissue of Patients

    Science.gov (United States)

    Jansson, V.; Giurea, A.

    2016-01-01

    Introduction. Increasing interest developed in the use of carbon-fiber-reinforced-poly-ether-ether-ketones (CFR-PEEK) as an alternative bearing material in knee arthroplasty. The effects of CFR-PEEK wear in in vitro and animal studies are controversially discussed, as there are no data available concerning human tissue. The aim of this study was to analyze human tissue containing CFR-PEEK as well as UHMWPE wear debris. The authors hypothesized no difference between the used biomaterials. Methods and Materials. In 10 patients during knee revision surgery of a rotating-hinge-knee-implant-design, synovial tissue samples were achieved (tibial inserts: UHMWPE; bushings and flanges: CFR-PEEK). One additional patient received revision surgery without any PEEK components as a control. The tissue was paraffin-embedded, sliced into 2 μm thick sections, and stained with hematoxylin and eosin in a standard process. A modified panoptical staining was also done. Results. A “wear-type” reaction was seen in the testing and the control group. In all samples, the UHMWPE particles were scattered in the tissue or incorporated in giant cells. CFR-PEEK particles were seen as conglomerates and only could be found next to vessels. CFR-PEEK particles showed no giant-cell reactions. In conclusion, the hypothesis has to be rejected. UHMWPE and PEEK showed a different scatter-behavior in human synovial tissue. PMID:27766256

  11. SEM investigation of heart tissue samples

    Energy Technology Data Exchange (ETDEWEB)

    Saunders, R; Amoroso, M [Physics Department, University of the West Indies, St. Augustine, Trinidad and Tobago, West Indies (Trinidad and Tobago)

    2010-07-01

    We used the scanning electron microscope to examine the cardiac tissue of a cow (Bos taurus), a pig (Sus scrofa), and a human (Homo sapiens). 1mm{sup 3} blocks of left ventricular tissue were prepared for SEM scanning by fixing in 96% ethanol followed by critical point drying (cryofixation), then sputter-coating with gold. The typical ridged structure of the myofibrils was observed for all the species. In addition crystal like structures were found in one of the samples of the heart tissue of the pig. These structures were investigated further using an EDVAC x-ray analysis attachment to the SEM. Elemental x-ray analysis showed highest peaks occurred for gold, followed by carbon, oxygen, magnesium and potassium. As the samples were coated with gold for conductivity, this highest peak is expected. Much lower peaks at carbon, oxygen, magnesium and potassium suggest that a cystallized salt such as a carbonate was present in the tissue before sacrifice.

  12. Histopathologic findings in uterine biopsy samples from subfertile bitches: 399 cases (1990-2005).

    Science.gov (United States)

    Gifford, Angela T; Scarlett, Janet M; Schlafer, Donald H

    2014-01-15

    To determine the prevalence of various lesion types detected by histologic evaluation of uterine biopsy samples collected from subfertile bitches. Retrospective case series. 399 sexually intact bitches. Results of histologic evaluation of canine uterine biopsy samples submitted by a single veterinary practice and clinical histories of dogs from which samples were obtained were reviewed. Clinical data including age, reason for biopsy, and histopathologic findings were recorded. The prevalence of specific lesions was determined, categorized by severity and age, and statistically analyzed. Endometritis (170/399 [42.6%] cases) and cystic endometrial changes, including cystic endometrial hyperplasia (133/399 [33.3%]) were the most prevalent lesions in the study population. Eighty-nine of 170 (52.4%) cases of endometritis were characterized as chronic with predominantly lymphocytic or lymphoplasmacytic inflammatory infiltrates, 51 (30.0%) included mixed inflammatory reactions, and 30 (17.6%) were characterized as having acute inflammation with neutrophils, eosinophils, or both. Fibrosis was common (101/399 [25.3%] cases). Eosinophilic endometritis was significantly associated with a history of fetal loss during the same breeding cycle. No significant difference was found in prevalence of lesions among age groups. The high prevalence of endometritis in this population of dogs suggested that acute and chronic endometritis may be related to subfertility in bitches. The association of eosinophilic endometrial infiltrates with a history of fetal loss may be an important diagnostic finding in dogs with endometritis.

  13. Histopathological study of the kidney, liver and intestine tissues in goldfish (Carassius auratus and angelfish (Pterophyllum sp.

    Directory of Open Access Journals (Sweden)

    Ebtesam Ahmadmoradi

    2012-09-01

    Full Text Available Histopathology is used for diagnosis of diseases in aquatic animals. This study was conductedto investigate histopathological lesions of kidney, liver and intestine of goldfish (Carassius auratus andangelfish (Pterophyllum sp. in some aquarium shops during a period between January-April 2010.Several histological alterations were observed in kidney which include glomerulonephritis, cell swelling ofepithelial cells, tubular epithelium necrosis, hyaline droplets and hyaline cast in renal tubules and tubulardilation. Liver showed vacuolar degeneration of hepatocytes, karyolysis and karyorrhexis and focal areasof necrosis, haemorrhagia and hepatitis. In intestine, atrophy of epithelial cells and enteritis on thelamina propria and submucosal layer were seen. It was concluded that unfavourable environmentalcontamination of aquarium may induced several histopathological alterations in the tissues of goldfishand angelfishes.

  14. Effect of dietary selenomethionine on growth performance, tissue burden, and histopathology in green and white sturgeon.

    Science.gov (United States)

    De Riu, Nicola; Lee, Jang-Won; Huang, Susie S Y; Moniello, Giuseppe; Hung, Silas S O

    2014-03-01

    A comparative examination of potential differences in selenium (Se) sensitivity was conducted on two sturgeon species indigenous to the San Francisco Bay-Delta. Juvenile green (Acipenser medirostris), recently given a federally threatened status, and white sturgeon (Acipenser transmontanus) were exposed to one of four nominal concentrations of dietary l-selenomethionine (SeMet) (0 (control), 50, 100, or 200 mg SeMet/kg diet) for 8 weeks. Mortality, growth performance, whole body composition, histopathology, and Se burdens of the whole body, liver, kidneys, gills, heart, and white muscle were determined every 2 to 4 weeks. Significant (psturgeon fed the highest SeMet diet after 2 weeks, whereas no mortality was observed in white sturgeon. Growth rates were significantly reduced in both species; however, green sturgeon was more adversely affected by the treatment. Dietary SeMet significantly affected whole body composition and most noticeably, in the decline of lipid contents in green sturgeon. Selenium accumulated significantly in all tissues relative to the control groups. After 4 and 8 weeks of exposure, marked abnormalities were observed in the kidneys and liver of both sturgeon species; however, green sturgeon was more susceptible to SeMet than white sturgeon at all dietary SeMet levels. Our results showed that a dietary Se concentration at 19.7 ± 0.6 mg Se/kg, which is in range with the reported Se concentrations of the benthic macro-vertebrate community of the San Francisco Bay, had adverse effects on both sturgeon species. However, the exposure had a more severe pathological effect on green sturgeon, suggesting that when implementing conservation measures, this federally listed threatened species should be monitored and managed independently from white sturgeon.

  15. Furcation Perforation: Periradicular Tissue Response to Biodentine as a Repair Material by Histopathologic and Indirect Immunofluorescence Analyses.

    Science.gov (United States)

    Silva, Lea Assed Bezerra; Pieroni, Karina Alessandra Michelão Grecca; Nelson-Filho, Paulo; Silva, Raquel Assed Bezerra; Hernandéz-Gatón, Patrícia; Lucisano, Marília Pacífico; Paula-Silva, Francisco Wanderley Garcia; de Queiroz, Alexandra Mussolino

    2017-07-01

    The purpose of this study was to evaluate the in vivo response of periradicular tissues after sealing of furcation perforations with Biodentine, mineral trioxide aggregate (MTA), and gutta-percha by means of histopathologic and indirect immunofluorescence analyses. Thirty teeth of 3 dogs were divided into 3 groups: Biodentine (n = 14 teeth), MTA (negative control, n = 10 teeth), and gutta-percha (positive control, n = 6 teeth). After endodontic treatment, perforations were made on the center of the pulp chamber floor and filled with the materials. After 120 days, the animals were killed, and blocks containing the teeth and periradicular tissues were processed histotechnically for histopathologic semiquantitative (new mineralized tissue formation and bone resorption at the perforation site) and quantitative (thickness and area of newly formed mineralized tissue and number of inflammatory cells) analyses and RUNX2 immunofluorescence assay. Data were analyzed by χ(2), Fisher exact test, Mann-Whitney test, one-way analysis of variance, Kruskal-Wallis test, and Dunn posttest (α = 0.05). MTA and Biodentine induced the formation of significantly more new mineralized tissue (P < .0001) than gutta-percha, which did not induce the formation of mineralized tissue in any case. Complete sealing of the perforations was more frequent with MTA, which formed mineralized tissue with greater thickness and area. Biodentine and MTA exhibited no bone resorption in the furcation region, fewer inflammatory cells, and greater RUNX2 immunostaining intensity than gutta-percha. Although MTA presented higher frequency of complete sealing and greater thickness and area of newly formed mineralized tissue, Biodentine also had good histopathologic results and can be considered as an adequate furcation perforation repair material. Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  16. Avian bornavirus is present in many tissues of psittacine birds with histopathologic evidence of proventricular dilatation disease.

    Science.gov (United States)

    Raghav, Raj; Taylor, Michael; Delay, Josepha; Ojkic, Davor; Pearl, David L; Kistler, Amy L; Derisi, Joseph L; Ganem, Donald; Smith, Dale A

    2010-07-01

    Proventricular dilatation disease (PDD) is a neurologic disease of psittacine birds suspected to be caused by a recently identified Avian bornavirus (ABV). In the current report, data supporting the causal association of ABV with PDD are presented. Immunohistochemistry (IHC) with rabbit polyclonal antiserum raised against ABV nucleocapsid protein was used to identify cell and organ distribution of viral antigen. The ABV antigen was most consistently detected in brain, spinal cord, adrenal gland, pancreas, and kidney. Histopathologic evaluation was correlated with ABV-specific polymerase chain reaction (PCR) and immunohistochemical tests in multiple tissues from 16 psittacine birds with and without PDD. Using histopathologic diagnosis as the gold standard, the sensitivity and specificity of IHC for ABV antigens were found to be 100% and 100%, respectively. Many more tissues were positive for ABV RNA by reverse transcription PCR than were positive for pathologic changes or viral antigens by IHC, indicating the presence of subclinical or asymptomatic infection at many sites.

  17. Multispectral Enhancement Method to Increase the Visual Differences of Tissue Structures in Stained Histopathology Images

    Directory of Open Access Journals (Sweden)

    Pinky A. Bautista

    2012-01-01

    Full Text Available In this paper we proposed a multispectral enhancement scheme in which the spectral colors of the stained tissue-structure of interest and its background can be independently modified by the user to further improve their visualization and color discrimination. The colors of the background objects are modified by transforming their N-band spectra through an NxN transformation matrix, which is derived by mapping the representative samples of their original spectra to the spectra of their target colors using least mean square method. On the other hand, the color of the tissue structure of interest is modified by modulating the transformed spectra with the sum of the pixel’s spectral residual-errors at specific bands weighted through an NxN weighting matrix; the spectral error is derived by taking the difference between the pixel’s original spectrum and its reconstructed spectrum using the first M dominant principal component vectors in principal component analysis. Promising results were obtained on the visualization of the collagen fiber and the non-collagen tissue structures, e.g., nuclei, cytoplasm and red blood cells (RBC, in a hematoxylin and eosin (H&E stained image.

  18. The effect of zinc deficiency and zinc supplementation on element levels in the bone tissue of ovariectomized rats: histopathologic changes.

    Science.gov (United States)

    Baltaci, Abdulkerim Kasim; Sunar, Fusun; Mogulkoc, Rasim; Acar, Musa; Toy, Hatice

    2014-05-01

    Study aimed to determine the effects of zinc supplementation/deficiency on the histological structure and elements levels in bone tissue in ovariectomized rats. The study included 40 Sprague-Dawley type adult female rats, divided as follows: Control, ovariectomized, ovariectomized + zinc supplemented, ovariectomized + zinc deficient groups. At the end of the study bone tissues (femur) were collected to determine the levels of calcium, phosphorus, magnesium, zinc, iron, aluminium, chrome, lithium, lead, nickel, and manganese. The bone tissue was examined for histopathology. Ovariectomy leaded to significant decrease in magnesium. Zinc supplementation to ovariectomized rats restored the reduced calcium, phosphorus, zinc. However, zinc deficiency in ovariectomized rats further reduced calcium, phosphorus, zinc, and manganese levels. Zinc deficiency in ovariectomized significantly increased Al, Cr, Li, Pb, and Ni levels. Tissue integrity was impaired due to ovariectomy and zinc deficiency. Ovariectomy and zinc deficiency leads significant decreases elements of the bone.

  19. Recent technological developments: in situ histopathological interrogation of surgical tissues and resection margins

    Directory of Open Access Journals (Sweden)

    Upile Tahwinder

    2007-03-01

    Full Text Available Abstract Objectives The tumour margin is an important surgical concept significantly affecting patient morbidity and mortality. We aimed in this prospective study to apply the microendoscope on tissue margins from patients undergoing surgery for oral cancer in vivo and ex vivo and compare it to the gold standard "paraffin wax", inter-observer agreement was measured; also to present the surgical pathologist with a practical guide to the every day use of the microendoscope both in the clinical and surgical fields. Materials and methods Forty patients undergoing resection of oral squamous cell carcinoma were recruited. The surgical margin was first marked by the operator followed by microendoscopic assessment. Biopsies were taken from areas suggestive of close or positive margins after microendoscopic examination. These histological samples were later scrutinized formally and the resection margins revisited accordingly when necessary. Results Using the microendoscope we report our experience in the determination of surgical margins at operation and later comparison with frozen section and paraffin section margins "gold standard". We were able to obtain a sensitivity of 95% and a specificity of 90%. Inter-observer Kappa scores comparing the microendoscope with formal histological analysis of normal and abnormal mucosa were 0.85. Conclusion The advantage of this technique is that a large area of mucosa can be sampled and any histomorphological changes can be visualized in real time allowing the operator to make important informed decisions with regards the intra-operative resection margin at the time of the surgery.

  20. A Systematic Approach to the Application of Soft Tissue Histopathology in Paleopathology

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    Christina Grove

    2015-01-01

    Full Text Available The application of histology to soft tissue remains offers an important technique to obtain diagnostically important information on various physiological and pathological conditions in paleopathology. In a series of 29 cases with mummified tissue ranging between 16 months and c. 5.200 years of postmortem time interval, we systematically investigated paleohistology and the preservation of various tissues. We established a reproducible histological ranking system for the evaluation of mummified tissue preservation. The application of this scheme to the series showed good tissue preservation of tissues with high connective tissue content but also fat tissue and connective tissue rich organs, such as lung tissue, while most other internal organs were less well preserved despite highly different postmortem time intervals. There are some organs with only poor conservation even in short term periods such as the kidneys and CNS. Artificial mummification does not provide better conservation than naturally mummified tissues; “cold” mummies may be much better conserved than those from desert areas. The identification of specific pathologies underlines the potential power of paleohistology.

  1. Breast cancer histopathology image analysis

    NARCIS (Netherlands)

    Veta, M.

    2014-01-01

    Pathology labs are currently undergoing a transformation towards a fully digital workflow. In addition to the digital management of tissue samples, pathology orders and reports, this includes the digitization of histopathology slides and use of computer monitors for viewing them, which aims to

  2. Breast cancer histopathology image analysis

    NARCIS (Netherlands)

    Veta, M.

    2014-01-01

    Pathology labs are currently undergoing a transformation towards a fully digital workflow. In addition to the digital management of tissue samples, pathology orders and reports, this includes the digitization of histopathology slides and use of computer monitors for viewing them, which aims to repla

  3. Improved histopathological evaluation of gliomas using tissue fragments obtained by ultrasonic aspiration

    DEFF Research Database (Denmark)

    Neckelmann, K; Kristensen, B W; Schrøder, H D

    2004-01-01

    the amount of tissue fragments, which need to be analyzed, and in order to save tissue for other purposes like molecular genetics and in vitro drug sensitivity assays. Thirty cases were included in the present study and consisted of astrocytomas, glioblastomas, oligodendrogliomas and ependymomas. The results...... the amount of slides, which have to be analyzed and saving Sonocut tissue fragments for future use in molecular genetics and drug sensitivity assays....

  4. Improved histopathological evaluation of gliomas using tissue fragments obtained by ultrasonic aspiration

    DEFF Research Database (Denmark)

    Neckelmann, K; Kristensen, B W; Schrøder, H D

    2004-01-01

    Ultrasonic aspirators are commonly used to resect brain tumors because they allow safe, rapid and accurate removal of dissected tissue. However, the tissue fragments removed by ultrasonic aspirators are used surprisingly little in daily diagnostics and in clinical research. A comparison between d...

  5. Histopathological examination of nerve samples from pure neural leprosy patients: obtaining maximum information to improve diagnostic efficiency

    Directory of Open Access Journals (Sweden)

    Sérgio Luiz Gomes Antunes

    2012-03-01

    Full Text Available Nerve biopsy examination is an important auxiliary procedure for diagnosing pure neural leprosy (PNL. When acid-fast bacilli (AFB are not detected in the nerve sample, the value of other nonspecific histological alterations should be considered along with pertinent clinical, electroneuromyographical and laboratory data (the detection of Mycobacterium leprae DNA with polymerase chain reaction and the detection of serum anti-phenolic glycolipid 1 antibodies to support a possible or probable PNL diagnosis. Three hundred forty nerve samples [144 from PNL patients and 196 from patients with non-leprosy peripheral neuropathies (NLN] were examined. Both AFB-negative and AFB-positive PNL samples had more frequent histopathological alterations (epithelioid granulomas, mononuclear infiltrates, fibrosis, perineurial and subperineurial oedema and decreased numbers of myelinated fibres than the NLN group. Multivariate analysis revealed that independently, mononuclear infiltrate and perineurial fibrosis were more common in the PNL group and were able to correctly classify AFB-negative PNL samples. These results indicate that even in the absence of AFB, these histopathological nerve alterations may justify a PNL diagnosis when observed in conjunction with pertinent clinical, epidemiological and laboratory data.

  6. Histopathological examination of nerve samples from pure neural leprosy patients: obtaining maximum information to improve diagnostic efficiency.

    Science.gov (United States)

    Antunes, Sérgio Luiz Gomes; Chimelli, Leila; Jardim, Márcia Rodrigues; Vital, Robson Teixeira; Nery, José Augusto da Costa; Corte-Real, Suzana; Hacker, Mariana Andréa Vilas Boas; Sarno, Euzenir Nunes

    2012-03-01

    Nerve biopsy examination is an important auxiliary procedure for diagnosing pure neural leprosy (PNL). When acid-fast bacilli (AFB) are not detected in the nerve sample, the value of other nonspecific histological alterations should be considered along with pertinent clinical, electroneuromyographical and laboratory data (the detection of Mycobacterium leprae DNA with polymerase chain reaction and the detection of serum anti-phenolic glycolipid 1 antibodies) to support a possible or probable PNL diagnosis. Three hundred forty nerve samples [144 from PNL patients and 196 from patients with non-leprosy peripheral neuropathies (NLN)] were examined. Both AFB-negative and AFB-positive PNL samples had more frequent histopathological alterations (epithelioid granulomas, mononuclear infiltrates, fibrosis, perineurial and subperineurial oedema and decreased numbers of myelinated fibres) than the NLN group. Multivariate analysis revealed that independently, mononuclear infiltrate and perineurial fibrosis were more common in the PNL group and were able to correctly classify AFB-negative PNL samples. These results indicate that even in the absence of AFB, these histopathological nerve alterations may justify a PNL diagnosis when observed in conjunction with pertinent clinical, epidemiological and laboratory data.

  7. Comparison of histopathology, cultivation of tissues and rectal contents, and interferon-gamma and serum antibody responses for the diagnosis of bovine paratuberculosis

    DEFF Research Database (Denmark)

    Huda, A.; Jensen, H.E.

    2003-01-01

    , hepatic and supramammary). Histopathological examination and bacterial culture of tissues each revealed that six of the 16 cattle were infected, whereas repeated culture of rectal contents revealed only five infected animals. Except for the hepatic lymph node of a single animal, culture-positive tissues...

  8. Protective effects of protocatechuic acid on TCDD-induced oxidative and histopathological damage in the heart tissue of rats.

    Science.gov (United States)

    Ciftci, Osman; Disli, Olcay Murat; Timurkaan, Necati

    2013-10-01

    2,3,7,8-Tetracholorodibenzo-p-dioxin (TCDD) is a highly toxic environmental contaminant that causes severe toxic effects in animal and human. In this study, we investigated the toxic effects of TCDD and the preventive effects of protocatechuic acid (PCA), a widespread phenolic compound, in the heart tissue of rats. For this purpose, 3-4 months old 28 rats with 280-310 g body weights were equally divided into 4 groups (control, TCDD, PCA, TCDD + PCA group). A 2 μg/kg dose of 2,3,7,8-TCDD and 100 mg/kg dose of PCA were dissolved in corn oil and given orally to the rats for 45 days. The results indicated that TCDD induced oxidative stress by increasing the level of thiobarbituric acid reactive substance and by decreasing the levels of glutathione, catalase, glutathione peroxidase and superoxide dismutase in the heart tissue of rats. In contrast, PCA treatment prevents the toxic effects of TCDD on oxidative stress. In addition, histopathological alterations such as necrosis and hemorrhage occurred in TCDD group, and PCA treatment partially prevents these alterations in heart tissue. In this study, it was concluded that TCDD exposure led to toxic effects in heart tissue and PCA treatment could prevent the toxicity of TCDD.

  9. Radioprotective effects of hesperidin on oxidative damages and histopathological changes induced by X-irradiation in rats heart tissue

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    Abolhasan Rezaeyan

    2016-01-01

    Full Text Available This study was carried out to evaluate radioprotective effects of hesperidin (HES administration before the irradiation on the cardiac oxidative stress and histopathological changes in an experimental rat model. The cardiovascular complications of radiation exposure cause morbidity and mortality in patients who received radiotherapy. HES, an antioxidant flavonoid found in citrus fruits, suggests the protection against the tissue damage. Fifty-eight rats were divided into four groups: Group 1 received phosphate buffered saline (PBS and sham radiation; Group 2, HES and sham radiation; Group 3, PBS and radiation; and Group 4, HES and radiation. The rats were exposed to single dose of 18 Gy of 6 MV X-ray. One hundred milligrams per kilogram doses of HES was administered for 7 days before irradiation. The estimation of superoxide dismutase (SOD, malondialdehyde (MDA, and histopathological analyses was performed at 24 h and 8 weeks after radiation exposure. The irradiation of chest area resulted in an elevated MDA level and decreased SOD activity. Moreover, long-term pathological lesions of radiation were inflammation, fibrosis, the increased number of mast cells and macrophages, and development of plaque, vascular leakage, myocardial degeneration, and myocyte necrosis. Although the administration of HES decreases inflammation, fibrosis, mast cell and macrophage numbers, and myocyte necrosis, it did not result in reduced thrombus, myocardium degeneration, and vascular leakage. In conclusion, these results suggest that HES can perform a radioprotection action. The protective effect of HES may be attributable to its immunomodulatory effects and free radical-scavenging properties.

  10. Histopathological changes induced by paraquat on some tissues of gourami fish (Trichogaster trichopterus

    Directory of Open Access Journals (Sweden)

    M. Banaee

    2013-03-01

    Full Text Available Paraquat is a contact and non-selective herbicide which is used for controlling a wide range of terrestrial weeds and aquatic plants. A long-term contact with this xenobiotic can potentially lead to injuries in fishes as live non-target organisms. Therefore, the current study aimed to investigate the effect of sub-lethal toxicity of paraquat on the pathology of gill, liver, and spleen tissues in gourami fish (Trichogaster trichopterus. In this study, sub-lethal concentration is determined based on lethal concentration (LC50 : 7.16±0.69, 4.46±0.43, 2.19±0.27 and 1.41±0.17 mg/l of paraquat within 24, 48, 72 and 96 hours, respectively. The experiment was done with four varied concentrations of paraquat (0.0, 0.07, 0.15, and 0.3 mg/l equal 0.0%, 5%, 10% and 20% of nominal value of 96 h LC50 during 3 weeks. The exposed fish displayed erratic swimming and became lethargic. The changes in gills were characterized by hypertrophy, epithelial, epithelium increase of gill filament, edema and secondary gill lamella. The liver showed hypotrophy of liver cells, cloudy swelling and formation of cytoplasmic vacuoles in the liver tissue of fish treated with 0.15 and 0.3 mg/l concentrations of paraquat. Disorder in the ellipsoid cell and hemosiderin accumulation in melano-macrophage centers was observed in the spleen tissue of fish exposed to 0.15 and 0.3 mg/l of paraquat.

  11. Multimodal Raman-fluorescence spectroscopy of formalin fixed samples is able to discriminate brain tumors from dysplastic tissue

    Science.gov (United States)

    Anand, Suresh; Cicchi, Riccardo; Giordano, Flavio; Buccoliero, Anna Maria; Pavone, Francesco Saverio

    2014-05-01

    In the recent years, there has been a considerable surge in the application of spectroscopy for disease diagnosis. Raman and fluorescence spectra provide characteristic spectral profile related to biochemical and morphological changes when tissues progress from normal state towards malignancy. Spectroscopic techniques offer the advantage of being minimally invasive compared to traditional histopathology, real time and quantitative. In biomedical optical diagnostics, freshly excised specimens are preferred for making ex-vivo spectroscopic measurements. With regard to fresh tissues, if the lab is located far away from the clinic it could pose a problem as spectral measurements have to be performed immediately after dissection. Tissue samples are usually placed in a fixative agent such as 4% formaldehyde to preserve the samples before processing them for routine histopathological studies. Fixation prevents the tissues from decomposition by arresting autolysis. In the present study, we intend to investigate the possibility of using formalin fixed samples for discrimination of brain tumours from dysplastic tissue using Raman and fluorescence spectroscopy. Formalin fixed samples were washed with phosphate buffered saline for about 5 minutes in order to remove the effects of formalin during spectroscopic measurements. In case of fluorescence spectroscopy, changes in spectral profile have been observed in the region between 550-670 nm between dysplastic and tumor samples. For Raman measurements, we found significant differences in the spectral profiles between dysplasia and tumor. In conclusion, formalin fixed samples can be potentially used for the spectroscopic discrimination of tumor against dysplastic tissue in brain samples.

  12. Performance assessment of automated tissue characterization for prostate H and E stained histopathology

    Science.gov (United States)

    DiFranco, Matthew D.; Reynolds, Hayley M.; Mitchell, Catherine; Williams, Scott; Allan, Prue; Haworth, Annette

    2015-03-01

    Reliable automated prostate tumor detection and characterization in whole-mount histology images is sought in many applications, including post-resection tumor staging and as ground-truth data for multi-parametric MRI interpretation. In this study, an ensemble-based supervised classification algorithm for high-resolution histology images was trained on tile-based image features including histogram and gray-level co-occurrence statistics. The algorithm was assessed using different combinations of H and E prostate slides from two separate medical centers and at two different magnifications (400x and 200x), with the aim of applying tumor classification models to new data. Slides from both datasets were annotated by expert pathologists in order to identify homogeneous cancerous and non-cancerous tissue regions of interest, which were then categorized as (1) low-grade tumor (LG-PCa), including Gleason 3 and high-grade prostatic intraepithelial neoplasia (HG-PIN), (2) high-grade tumor (HG-PCa), including various Gleason 4 and 5 patterns, or (3) non-cancerous, including benign stroma and benign prostatic hyperplasia (BPH). Classification models for both LG-PCa and HG-PCa were separately trained using a support vector machine (SVM) approach, and per-tile tumor prediction maps were generated from the resulting ensembles. Results showed high sensitivity for predicting HG-PCa with an AUC up to 0.822 using training data from both medical centres, while LG-PCa showed a lower sensitivity of 0.763 with the same training data. Visual inspection of cancer probability heatmaps from 9 patients showed that 17/19 tumors were detected, and HG-PCa generally reported less false positives than LG-PCa.

  13. Molecular and Histopathological Changes in Mouse Intestinal Tissue After Proton Exposure

    Science.gov (United States)

    Purgason, Ashley; Wu, Honglu

    2010-01-01

    Whole body exposure to protons in mice causes significant apoptosis in the crypts of the small intestine. Increasing numbers of crypts contained more apoptotic lesions as the dose of exposure increased. 16 genes associated with apoptotic pathways were shown to have significantly altered expression as compared to control samples for at least one of the doses of proton exposure 1 gene, Trp53inp1, was significantly up-regulated across all three doses. Those animals exposed to 0.1 Gy of proton irradiation showed greater amounts of significant alterations in gene expression as compared to 1 Gy and 2 Gy exposures. The differences in gene expression changes of low and high dose proton irradiated mice may offer insight into the molecular mechanisms of the possible high sensitivity at low proton doses. RAIDD (CRADD) may be responsible for the hypersensitivity observed in the duodenum of mice exposed to low doses of protons. Caspase-1 may also play a role in the hypersensitivity seen following proton irradiation at a dose of 0.1 Gy. FOXO3A may be involved in the down-regulation of GILZ observed at high doses of proton exposure.

  14. Organochlorine-induced histopathology in kidney and liver tissue from Arctic fox (Vulpes lagopus).

    Science.gov (United States)

    Sonne, Christian; Wolkers, Hans; Leifsson, Pall S; Jenssen, Bjørn Munro; Fuglei, Eva; Ahlstrøm, Oystein; Dietz, Rune; Kirkegaard, Maja; Muir, Derek C G; Jørgensen, Even

    2008-04-01

    The effects of persistent organic pollutants on renal and liver morphology in farmed arctic fox (Vulpes lagopus) were studied under experimental conditions. Control animals received a diet containing pork (Sus scrofa) fat with low amounts of persistent organic pollutants, while the diet of the exposed animals contained whale blubber, 'naturally' contaminated with persistent organic pollutants. Polychlorinated biphenyls (PCB) and organochlorine pesticide (OCP) concentrations in the whale blubber were 488 and 395 ng/g wet weight, respectively. Animals were sacrificed and sampled when they were at their fattest (winter) as well as their lowest body weight (summer). The results show that PCB and OCP exposure causes renal (and probably also liver) lesions in arctic foxes. The prevalence of glomerular, tubular and interstitial lesions was significantly highest in the exposed group (chi-square: all p0.05). The prevalence of lesions was not significantly different between lean (winter) and fat (summer) foxes for any of the lesions (chi-square: all p>0.05). We suggest that wild arctic foxes exposed to an environmental cocktail of persistent organic pollutants, such as PCBs and OCPs, in their natural diet are at risk for developing chronic kidney and liver damage. Whether such lesions may have an impact on age and health of the animals remains uncertain.

  15. A Comparison of Molecular and Histopathological Changes in Mouse Intestinal Tissue Following Whole-Body Proton- or Gamma-Irradiation

    Science.gov (United States)

    Purgason, Ashley; Mangala, Lingegowda; Zhang, Ye; Hamilton, Stanley; Wu, Honglu

    2010-01-01

    There are many consequences following exposure to the space radiation environment which can adversely affect the health of a crew member. Acute radiation syndrome (ARS) involving nausea and vomiting, damage to radio-sensitive tissue such as the blood forming organs and gastrointestinal tract, and cancer are some of these negative effects. The space radiation environment is ample with protons and contains gamma rays as well. Little knowledge exists to this point, however, regarding the effects of protons on mammalian systems; conversely several studies have been performed observing the effects of gamma rays on different animal models. For the research presented here, we wish to compare our previous work looking at whole-body exposure to protons using a mouse model to our studies of mice experiencing whole-body exposure to gamma rays as part of the radio-adaptive response. Radio-adaptation is a well-documented phenomenon in which cells exposed to a priming low dose of radiation prior to a higher dose display a reduction in endpoints like chromosomal aberrations, cell death, micronucleus formation, and more when compared to their counterparts receiving high dose-irradiation only. Our group has recently completed a radio-adaptive experiment with C57BL/6 mice. For both this study and the preceding proton research, the gastrointestinal tract of each animal was dissected four hours post-irradiation and the isolated small intestinal tissue was fixed in formalin for histopathological examination or snap-frozen in liquid nitrogen for RNA isolation. Histopathologic observation of the tissue using standard H&E staining methods to screen for morphologic changes showed an increase in apoptotic lesions for even the lowest doses of 0.1 Gy of protons and 0.05 Gy of gamma rays, and the percentage of apoptotic cells increased with increasing dose. A smaller percentage of crypts showed 3 or more apoptotic lesions in animals that received 6 Gy of gamma-irradiation compared to mice

  16. A texture based pattern recognition approach to distinguish melanoma from non-melanoma cells in histopathological tissue microarray sections.

    Directory of Open Access Journals (Sweden)

    Elton Rexhepaj

    Full Text Available AIMS: Immunohistochemistry is a routine practice in clinical cancer diagnostics and also an established technology for tissue-based research regarding biomarker discovery efforts. Tedious manual assessment of immunohistochemically stained tissue needs to be fully automated to take full advantage of the potential for high throughput analyses enabled by tissue microarrays and digital pathology. Such automated tools also need to be reproducible for different experimental conditions and biomarker targets. In this study we present a novel supervised melanoma specific pattern recognition approach that is fully automated and quantitative. METHODS AND RESULTS: Melanoma samples were immunostained for the melanocyte specific target, Melan-A. Images representing immunostained melanoma tissue were then digitally processed to segment regions of interest, highlighting Melan-A positive and negative areas. Color deconvolution was applied to each region of interest to separate the channel containing the immunohistochemistry signal from the hematoxylin counterstaining channel. A support vector machine melanoma classification model was learned from a discovery melanoma patient cohort (n = 264 and subsequently validated on an independent cohort of melanoma patient tissue sample images (n = 157. CONCLUSION: Here we propose a novel method that takes advantage of utilizing an immuhistochemical marker highlighting melanocytes to fully automate the learning of a general melanoma cell classification model. The presented method can be applied on any protein of interest and thus provides a tool for quantification of immunohistochemistry-based protein expression in melanoma.

  17. Investigation of biochemical and histopathological effects of Mentha piperita Labiatae and Mentha spicata Labiatae on liver tissue in rats.

    Science.gov (United States)

    Akdogan, M; Ozguner, M; Aydin, G; Gokalp, O

    2004-01-01

    The plant Mentha piperita, or peppermint, is commonly used in the treatment of loss of appetite, common cold, bronchitis, sinusitis, fever, nausea and vomiting, and indigestion as a herbal agent. In this study, we aimed to investigate biochemical and histological effects of M. piperita Labiatae, growing in the Yenisar Bademli town of Isparta city, and Mentha spicata Labiatae, growing in the Anamas high plateau of the Yenisar Bademli town, on the rat liver tissue. Forty-eight male Wistar albino rats weighing 200-250 g were used for this study. Rats were divided into four groups of 12 animals: Group I received no herbal tea (control group); Group II received 20 g/L M. piperita tea; Group III received 20 g/L M. spicata tea; and Group IV received 40 g/L M. spicata tea. Herbal teas were prepared daily and provided at all times to the rats during 30 days as drinking water. Liver function tests, including aspartate aminotransferase (AST/GOT) and alanine aminotransferase (ALT/GPT) activities were measured. To evaluate liver antioxidant defences, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) and thiobarbituric acid reactive substance (TBARS) activities were determined in the homogenates of liver tissue. In addition, liver tissues were submitted for histopathologic examination. AST and ALT activities were increased in Group II, Group III and Group IV gradually when compared with the control group. The difference between Group II and the control group was not statistically significant (P > 0.016). Increases in AST and ALT activities of Group III and Group IV were statistically significant when compared with the control group. SOD, GSH-Px and CAT activities were increased in Group II when compared with the control group but the difference was not statistically significant (P > 0.016). However, SOD, GSH-Px activities and the TBARS level were significantly increased, and CAT activity was significantly decreased in Group III when compared with the

  18. Histopathologic investigation of the protective effects of omega-3 fatty acids against boric acid-induced injury in kidney and testis tissue

    Directory of Open Access Journals (Sweden)

    1 Hacettepe Üniversitesi, Tıp Fakültesi, Histoloji ve Embriyoloji Anabilim Dalı, Ankara, Türkiye 2 Mustafa Kemal Üniversitesi, Tıp Fakültesi, Histoloji ve Embriyoloji Anabilim Dalı, Hatay, Türkiye 3 Mustafa Kemal Üniversitesi, Hatay Sağlık Hizmetleri Meslek Yüksek Okulu, Hatay , Türkiye 4 Antakya Doğumevi, Hatay, Türkiye 5 Fatih Üniversitesi, Tıp Fakültesi, Histoloji ve Embriyoloji Anabilim Dalı, İstanbul, Türkiye 6 Turgut Özal Üniversitesi, Sağlık Bilimleri Meslek Yüksek Okulu, Ankara, Türkiye Yazışma Adresi / Correspondence : Ahmet Nacar, Hacettepe Üniversitesi, Tıp Fakültesi, Histoloji ve Embriyoloji Anabilim Dalı, Ankara, Türkiye Email: drnacar@gmail.com Geliş Tarihi / Received: 09.01.2014, Kabul Tarihi / Accepted: 10.02.2014 Copyright © Dicle Tıp Dergisi 2014, Her hakkı saklıdır / All rights reserved Dicle Tıp Dergisi / 2014; 41 (2: 385-390 Dicle Medical Journal doi: 10.5798/diclemedj.0921.2014.02.0436 ÖZGÜN ARAŞTIRMA / ORIGINAL ARTICLE Borik asit uygulamasının sıçan böbrek ve testis dokusunda oluşturduğu hasara karşı Omega-3 yağ asitlerinin koruyucu etkisinin histopatolojik olarak incelenmesi Histopathologic investigation of the protective effects of omega-3 fatty acids against boric acid-induced injury in kidney and testis tissue Ahmet Nacar

    2014-06-01

    Full Text Available Objective: In this study, it was aimed to evaluate the effects of boric acid on rat kidney and testis tissues histopathologically. Secondly, the protective effects of omega-3 fatty acid against boric acid-induced renal and testicular toxicity were investigated. Methods: 32 wistar albino rats were divided into 4 groups as follows: Control, Omega-3 (400 mg/kg/day for 10 days, Boric acid (375 mg/kg/day for 10 days and Boric acid+omega-3 (both drugs same dosage for same day. Kidney and testis tissues were evaluated using a scoring system based on the extent of certain histopathological changes. Results: In histopathological examination, boric acid caused significant degeneration in both testis and kidney tissues. Most evident findings were glomerular shrinkage and necrosis, hemorrhage and tubular cell degeneration in kidneys, and exfoliation of seminiferous tubule cells, detachement of epithelium from basement membrane, decreased cellularity and degeneration in epithelial cells in testis tissues. Omega-3 administration significantly attenuated these changes. Conclusion: To our literature search, this is the first study reporting protective effects of omega-3 fatty acid against boric-acid-induced testicular and renal injury.

  19. Specimen Sample Preservation for Cell and Tissue Cultures

    Science.gov (United States)

    Meeker, Gabrielle; Ronzana, Karolyn; Schibner, Karen; Evans, Robert

    1996-01-01

    The era of the International Space Station with its longer duration missions will pose unique challenges to microgravity life sciences research. The Space Station Biological Research Project (SSBRP) is responsible for addressing these challenges and defining the science requirements necessary to conduct life science research on-board the International Space Station. Space Station will support a wide range of cell and tissue culture experiments for durations of 1 to 30 days. Space Shuttle flights to bring experimental samples back to Earth for analyses will only occur every 90 days. Therefore, samples may have to be retained for periods up to 60 days. This presents a new challenge in fresh specimen sample storage for cell biology. Fresh specimen samples are defined as samples that are preserved by means other than fixation and cryopreservation. The challenge of long-term storage of fresh specimen samples includes the need to suspend or inhibit proliferation and metabolism pending return to Earth-based laboratories. With this challenge being unique to space research, there have not been any ground based studies performed to address this issue. It was decided hy SSBRP that experiment support studies to address the following issues were needed: Fixative Solution Management; Media Storage Conditions; Fresh Specimen Sample Storage of Mammalian Cell/Tissue Cultures; Fresh Specimen Sample Storage of Plant Cell/Tissue Cultures; Fresh Specimen Sample Storage of Aquatic Cell/Tissue Cultures; and Fresh Specimen Sample Storage of Microbial Cell/Tissue Cultures. The objective of these studies was to derive a set of conditions and recommendations that can be used in a long duration microgravity environment such as Space Station that will permit extended storage of cell and tissue culture specimens in a state consistent with zero or minimal growth, while at the same time maintaining their stability and viability.

  20. Workflow for large-scale analysis of melanoma tissue samples

    Directory of Open Access Journals (Sweden)

    Maria E. Yakovleva

    2015-09-01

    Full Text Available The aim of the present study was to create an optimal workflow for analysing a large cohort of malignant melanoma tissue samples. Samples were lysed with urea and enzymatically digested with trypsin or trypsin/Lys C. Buffer exchange or dilution was used to reduce urea concentration prior to digestion. The tissue digests were analysed directly or following strong cation exchange (SCX fractionation by nano LC–MS/MS. The approach which resulted in the largest number of protein IDs involved a buffer exchange step before enzymatic digestion with trypsin and chromatographic separation in 120 min gradient followed by SCX–RP separation of peptides.

  1. Tissue sampling methods and standards for vertebrate genomics

    Directory of Open Access Journals (Sweden)

    Wong Pamela BY

    2012-07-01

    Full Text Available Abstract The recent rise in speed and efficiency of new sequencing technologies have facilitated high-throughput sequencing, assembly and analyses of genomes, advancing ongoing efforts to analyze genetic sequences across major vertebrate groups. Standardized procedures in acquiring high quality DNA and RNA and establishing cell lines from target species will facilitate these initiatives. We provide a legal and methodological guide according to four standards of acquiring and storing tissue for the Genome 10K Project and similar initiatives as follows: four-star (banked tissue/cell cultures, RNA from multiple types of tissue for transcriptomes, and sufficient flash-frozen tissue for 1 mg of DNA, all from a single individual; three-star (RNA as above and frozen tissue for 1 mg of DNA; two-star (frozen tissue for at least 700 μg of DNA; and one-star (ethanol-preserved tissue for 700 μg of DNA or less of mixed quality. At a minimum, all tissues collected for the Genome 10K and other genomic projects should consider each species’ natural history and follow institutional and legal requirements. Associated documentation should detail as much information as possible about provenance to ensure representative sampling and subsequent sequencing. Hopefully, the procedures outlined here will not only encourage success in the Genome 10K Project but also inspire the adaptation of standards by other genomic projects, including those involving other biota.

  2. Histopathologic and Flow-Cytometric Analysis of Neoplastic and Benign “background” Tissue in Breast Carcinoma Resections

    Directory of Open Access Journals (Sweden)

    Daniel W. Visscher

    1998-01-01

    Full Text Available Two-color, multiparametric synthesis phase fraction (SPF analysis of cytokeratin-labeled epithelial cells was flow cytometrically performed on both benign (SPFb and malignant tissue samples (if available, SPFt from 132 mastectomy/lumpectomy specimens. These data were then correlated with clinicopathologic features, including (1 tumor differentiation, (2 the proportion of tumor comprised of duct carcinoma-in situ (DCIS, and (3 the histology of accompanying benign breast tissue, classified by predominant microscopic pattern as intact, normal terminal duct lobular units (NTDLU, 34% of cases, atrophic (AT, 33% of cases, proliferative fibrocystic (PFC, 26% of cases, and non-proliferative fibrocystic (NPFC, 7% of cases. SPFt was inversely correlated with extent of DCIS (DCIS =0 – 20% tumor volume – 12.7% mean SPFt, vs. DCIS >20% tumor volume – 6.4% mean SPFt, p = 0.001. SPFt also correlated with the histology of background benign breast tissue (NTDLU – 14.8% mean SPFt vs. AT – 6.9% mean SPFt vs. PFC – 12.7% mean SPFt, p = 0.05 but it did not correlate with patient age or SPFb (overall mean =0.73%. SPFb was correlated with patient age (>56 yr – 0.59% mean SPFb vs. < yr – 0.84% mean SPFb, p = 0.02, with background histology (NTDLU – 1.1% mean SPFb vs. AT – 0.43% mean SPFb vs. PFC – 0.70% mean SPFb, p < 0.02 and with the grade of the neoplasm (well/moderate – 0.58% mean vs. poorly differentiated – 0.85% mean, p = 0.04. Patients having a background of PFC were significantly older than patients with a background of NTDLU (45.2 yr vs. 60.2 yr, p = 0.01.

  3. [PCR-based diagnosis of mucormycosis in tissue samples].

    Science.gov (United States)

    Bialek, R; Zelck, U E

    2013-11-01

    Mucormycosis is characterized by a rapid, often fatal progression. Early diagnosis of invasive mucormycosis is the key for timely therapeutic intervention and improved survival. Contrary to the more prevalent aspergillosis, effective antifungal therapy of mucormycosis is mainly limited to amphotericin B. Given the importance to guide the timely initiation of amphotericin B and possible surgical intervention, rapid and specific identification of fungal hyphae is essential. Conventional histopathology depends on abundance and morphology of the fungi as well as on the skills of the personnel, and usually shows an accuracy of 80 %. PCR assays targeting fungal ribosomal genes to identify Mucorales at least at genus level increase sensitivity, allow a rapid identification as well as detection of double mold infections. Thus, PCR assays are beneficial to complement existing approaches. They are recommended to rapidly specify tissue diagnosis and accurate identification of fungi. This will help to guide effective therapy and thereby, survival will increase. Retrospective analyses of mucormycosis by PCR help to evaluate therapeutic interventions and will optimize treatment options.

  4. Light propagation in tissues: effect of finite size of tissue sample

    Science.gov (United States)

    Melnik, Ivan S.; Dets, Sergiy M.; Rusina, Tatyana V.

    1995-12-01

    Laser beam propagation inside tissues with different lateral dimensions has been considered. Scattering and anisotropic properties of tissue critically determine spatial fluence distribution and predict sizes of tissue specimens when deviations of this distribution can be neglected. Along the axis of incident beam the fluence rate weakly depends on sample size whereas its relative increase (more than 20%) towards the lateral boundaries. The finite sizes were considered to be substantial only for samples with sizes comparable with the diameter of the laser beam. Interstitial irradiance patterns simulated by Monte Carlo method were compared with direct measurements in human brain specimens.

  5. Proteomic analysis of tissue samples in translational breast cancer research

    DEFF Research Database (Denmark)

    Gromov, Pavel; Moreira, José; Gromova, Irina

    2014-01-01

    , and both prognosis and prediction of outcome of chemotherapy. The purpose of this review is to critically appraise what has been achieved to date using proteomic technologies and to bring forward novel strategies - based on the analysis of clinically relevant samples - that promise to accelerate......In the last decade, many proteomic technologies have been applied, with varying success, to the study of tissue samples of breast carcinoma for protein expression profiling in order to discover protein biomarkers/signatures suitable for: characterization and subtyping of tumors; early diagnosis...

  6. Glypican 3 expression in human nonneoplastic, preneoplastic, and neoplastic tissues: a tissue microarray analysis of 4,387 tissue samples.

    Science.gov (United States)

    Baumhoer, Daniel; Tornillo, Luigi; Stadlmann, Sylvia; Roncalli, Massimo; Diamantis, Eva Karamitopoulou; Terracciano, Luigi M

    2008-06-01

    Several studies have shown that glypican 3 (GPC3) could be a useful diagnostic marker for hepatocellular carcinoma (HCC) and for differentiating HCC from nonneoplastic and preneoplastic liver disease. To systematically investigate the epidemiology of GPC3 expression in the liver and in other organs and tissues, we used tissue microarray technology comprising 4,387 tissue samples from 139 tumor categories and 36 nonneoplastic and preneoplastic tissue types. The immunohistochemical expression of GPC3 was assessed semiquantitatively using a 10% cutoff score and was detected in 9.2% of nonneoplastic liver samples (11/119), 16% of preneoplastic nodular liver lesions (6/38), and 63.6% of HCCs (140/220), underlining the role of GPC3 in hepatocarcinogenesis. Furthermore, several other tumors revealed consistent expression of GPC3, including squamous cell carcinoma of the lung (27/50 [54%]), testicular nonseminomatous germ cell tumors (32/62 [52%]), and liposarcoma (15/29 [52%]).

  7. Identification of fungal pathogens in Formalin-fixed, Paraffin-embedded tissue samples by molecular methods.

    Science.gov (United States)

    Rickerts, Volker

    2016-02-01

    The etiology of invasive fungal infections (IFI) is incompletely understood due to diagnostic limitations including insensitivity of cultures and failure of histopathology to discriminate between different species. This diagnostic gap precludes the optimal use of antifungals, leading to adverse patient outcomes. The identification of fungal pathogens from Formalin-fixed, Paraffin-embedded tissue (FFPE) blocks by molecular methods is emerging as an alternative approach to study the etiology of IFI. PCR assays, including species specific- and broadrange fungal tests are used with FFPE samples from patients with proven IFI. Fungal species identification is achieved in 15-90% of the samples. This heterogeneity may be explained by the samples studied. However, comparison of different studies is impaired, as controls ruling out false positive-, false negative test results or PCR inhibition are frequently not reported. Studies using in situ hybridization also vary in the clinical samples included and the targeted fungi. In addition, target sequences, the probe chemistry and the detection of hybridization signals also account for the differences in diagnostic sensitivity. Using both approaches in parallel yields additive insights, potentially leading to a superior identification of fungal etiology and awareness of the limitations of both molecular diagnostic approaches.

  8. Cell and tissue biomarkers in mussel, and histopathology in hake and anchovy from Bay of Biscay after the Prestige oil spill (Monitoring Campaign 2003)

    Energy Technology Data Exchange (ETDEWEB)

    Marigomez, Ionan [Zoologia eta Biologia Zelularra Saila, Zientzia eta Teknologia Fakultatea, Euskal Herriko Unibertsitatea, 644 P.K., E-48080 Bilbo, Basque Country (Spain)]. E-mail ionan.marigomez@ehu.es; Soto, Manu [Zoologia eta Biologia Zelularra Saila, Zientzia eta Teknologia Fakultatea, Euskal Herriko Unibertsitatea, 644 P.K., E-48080 Bilbo, Basque Country (Spain); Cancio, Ibon [Zoologia eta Biologia Zelularra Saila, Zientzia eta Teknologia Fakultatea, Euskal Herriko Unibertsitatea, 644 P.K., E-48080 Bilbo, Basque Country (Spain); Orbea, Amaia [Zoologia eta Biologia Zelularra Saila, Zientzia eta Teknologia Fakultatea, Euskal Herriko Unibertsitatea, 644 P.K., E-48080 Bilbo, Basque Country (Spain); Garmendia, Larraitz [Zoologia eta Biologia Zelularra Saila, Zientzia eta Teknologia Fakultatea, Euskal Herriko Unibertsitatea, 644 P.K., E-48080 Bilbo, Basque Country (Spain); Cajaraville, Miren P. [Zoologia eta Biologia Zelularra Saila, Zientzia eta Teknologia Fakultatea, Euskal Herriko Unibertsitatea, 644 P.K., E-48080 Bilbo, Basque Country (Spain)

    2006-07-01

    In order to assess the biological effects of the Prestige oil spill (POS), mussels (Mytilus galloprovincialis), European hake (Merluccius merluccius) and European anchovy (Engraulis encrasicolus) were sampled between April and September 2003 in various geographical areas of Bay of Biscay: Galicia, Central Cantabrian and East Cantabrian. In mussels, several cell and tissue biomarkers were measured: peroxisome proliferation as induction of acyl-CoA oxidase (AOX) activity, lysosomal responses as changes in the structure (lysosomal volume density, V{sub V{sub L}}, surface-to-volume ratio, S/V{sub L}, and numerical density, N{sub V{sub L}}) and in membrane stability (labilization period, LP), cell type replacement as relative proportion of basophilic cells (volume density of basophilic cells, V{sub V{sub BAS}}) in digestive gland epithelium, and changes in the morphology of digestive alveoli as mean luminal radius to mean epithelial thickness (MLR/MET). Additionally, flesh condition index (FCI) and gonad index (GI) were measured as supporting parameters. In hake and anchovy, liver histopathology was examined to determine the prevalence of parasites, melanomacrophage centers, non-specific lesions (inflammatory changes, atrophy, necrosis, apoptosis), early non-neoplastic toxicopathic lesions (i.e., hepatocellular nuclear polymorphism), foci of cellular alteration, benign and malignant neoplasms. In mussels, AOX induction was noticeable in April except in Galicia. LP values were low in all the geographical areas studied, indicating disturbed health, especially in Galicia. Alike, lysosomal enlargement was observed in most stations as shown by the extremely low S/V{sub L} values. V{sub V{sub BAS}} and MLR/MET values were markedly high. Overall, employed biomarkers detected exposure to toxic chemicals and disturbed health in mussels from Bay of Biscay, with impact decreasing from April to September. Although hepatocellular nuclear polymorphism and nematode parasitization in

  9. Theory of sampling and its application in tissue based diagnosis

    Directory of Open Access Journals (Sweden)

    Kayser Gian

    2009-02-01

    Full Text Available Abstract Background A general theory of sampling and its application in tissue based diagnosis is presented. Sampling is defined as extraction of information from certain limited spaces and its transformation into a statement or measure that is valid for the entire (reference space. The procedure should be reproducible in time and space, i.e. give the same results when applied under similar circumstances. Sampling includes two different aspects, the procedure of sample selection and the efficiency of its performance. The practical performance of sample selection focuses on search for localization of specific compartments within the basic space, and search for presence of specific compartments. Methods When a sampling procedure is applied in diagnostic processes two different procedures can be distinguished: I the evaluation of a diagnostic significance of a certain object, which is the probability that the object can be grouped into a certain diagnosis, and II the probability to detect these basic units. Sampling can be performed without or with external knowledge, such as size of searched objects, neighbourhood conditions, spatial distribution of objects, etc. If the sample size is much larger than the object size, the application of a translation invariant transformation results in Kriege's formula, which is widely used in search for ores. Usually, sampling is performed in a series of area (space selections of identical size. The size can be defined in relation to the reference space or according to interspatial relationship. The first method is called random sampling, the second stratified sampling. Results Random sampling does not require knowledge about the reference space, and is used to estimate the number and size of objects. Estimated features include area (volume fraction, numerical, boundary and surface densities. Stratified sampling requires the knowledge of objects (and their features and evaluates spatial features in relation to

  10. Comparison of histopathology and PCR based assay for detection of experimentally induced toxoplasmosis in murine model

    Institute of Scientific and Technical Information of China (English)

    Vikrant Sudan; A K Tewari; R Singh; Harkirat Singh

    2015-01-01

    Objective:To compare histopathology and PCR based detection in diagnosis of experimentally induced toxoplasmosis of RH human strain of the parasite in murine models. Methods:A comparison of histopathology and PCR based detection was done to diagnose experimentally induced toxoplasmosis in ten inbred swiss albino mice after intraperitoneal inoculation of 100 tachyzoites of laboratory mantained human RH strain of the parasite. Tissue samples from lung, liver, spleen, brain, heart and kidney were taken and processed for histopathological examination while all the samples also were subjected to PCR, using primers directed to the multicopy of SAG 3 gene, in dublicates. Results: Histopathology revealed presence of tachyzoites only in liver while along with lung, liver, spleen and brain tissue yielded desired positive PCR amplicons. Conclusions:The SAG 3 based PCR is able to diagnose toxoplasmosis in those tissues which are declared negative by histopathological assay.

  11. [Analysis of human tissue samples for volatile fire accelerants].

    Science.gov (United States)

    Treibs, Rudolf

    2014-01-01

    In police investigations of fires, the cause of a fire and the fire debris analysis regarding traces of fire accelerants are important aspects for forensic scientists. Established analytical procedures were recently applied to the remains of fire victims. When examining lung tissue samples, vapors inhaled from volatile ignitable liquids could be identified and differentiated from products of pyrolysis caused by the fire. In addition to the medico-legal results this evidence allowed to draw conclusions as to whether the fire victim was still alive when the fire started.

  12. Mercury and histopathology of the vulnerable goliath grouper, Epinephelus itajara, in U.S. waters: a multi-tissue approach.

    Science.gov (United States)

    Adams, Douglas H; Sonne, Christian

    2013-10-01

    Goliath grouper have undergone significant global population declines with potential biological extinction for some subpopulations. Although overfishing and habitat loss are important drivers of these declines, the negative effects of contaminants may also play a role. The life history patterns of goliath grouper may make this species especially prone to exposure to contaminants and may exacerbate bioaccumulation of toxic substances, including mercury, which has documented detrimental health effects. Therefore, we analyzed mercury (in muscle, liver, kidney, gonad, and brain tissue) and the histology of key organs (liver, kidney and gill tissue) in 56 goliath groupers from U.S. waters. Total mercury concentration was greatest in liver tissue, followed by kidney, muscle, gonad, and brain. Maximum mercury concentration ranged from 22.68 μg/g in liver tissue to 0.89 μg/g in brain tissue. Mean mercury concentration ranged from 2.87 μg/g in liver tissue to 0.37 μg/g in brain tissue with a mean of 0.63 μg/g in muscle. Mean mercury concentrations observed in goliath grouper from U.S. waters were within the range known to cause direct health effects in fish after long-term exposure. The lesions and histological changes observed in the liver, kidney, and gills of goliath groupers were similar to those found in other fish following laboratory mercury-exposure trials and to those found in mercury-contaminated fish in wild populations carrying similar or even lower concentrations. We suggest that exposure to mercury and other environmental influences such as pathogens and reduced temperatures could be co-factors in the histological effects or anomalies observed in the present study, and resulting stresses may be involved in the observed population declines in the species.

  13. Immunoelectron microscopic localization of elastic tissue components in archival tissue samples.

    Science.gov (United States)

    Fanning, J C; White, J F; Polewski, R; Cleary, E G

    1991-06-01

    Tissue samples that have been stored for many years, in different media and under a variety of conditions, have been examined by modern techniques of immunoelectron microscopy, using antibodies against elastic tissue components. A range of postembedding restorative procedures has been identified, which will allow reliable immunolocalization of antibodies against the elastic tissue component of such specimens. These methods have been applied successfully to autopsy-derived material, fixed in buffered formaldehyde, to archival material stored frozen at -70 or -20 degrees C, to specimens fixed for electron microscopy and stored for many years in buffer, and even to archival material from formaldehyde-fixed, paraffin-embedded blocks, reprocessed for electron microscopic examination. The successful restorative methods included pre-treatment of the sections with 6 M guanidine hydrochloride, or 1 M Tris/saline, each containing 100 mM dithiothreitol (a reducing agent) followed by alkylation with 220 mM iodoacetamide. The application of these techniques allowed reliable study of elastic tissue antibody distributions in archival tissues that could not be obtained again, as well as comparative studies with tissues processed many years previously.

  14. Demonstration of the histopathological and immunohistochemical effects of a novel hemostatic agent, ankaferd blood stopper, on vascular tissue in a rat aortic bleeding model

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    Gul Aylin

    2010-11-01

    Full Text Available Abstract Background Ankaferd Blood Stopper® (ABS is a folkloric medicinal plant extract used as a hemostatic agent in traditional Turkish medicine. This experimental study investigated the histopathological and immunohistochemical effects of ABS on vascular tissue in a rat model of aortic bleeding. Methods Four groups of 11 Wistar albino rats were used. The abdominal aortas of the rats were wounded; an ABS-soaked tampon was applied to rats in Groups 1 and 3, and a plain gauze tampon was applied to rats in Groups 2 and 4 until the bleeding stopped. The bleeding time was recorded. Immediately following sacrificing, the arteriotomy sites from Groups 1 and 2 were removed. The abdominal incisions in Groups 3 and 4 were closed following hemostasis. On Day 7 of the study, Group 3 and 4 rats were sacrificed and the abdominal aorta arteriotomy sites were removed for histopathological and immunohistochemical evaluation. Results The mean bleeding time in 15 animals in Groups 2 and 4 was 4.9 ± 0.6 s, and in 22 animals in Groups 1 and 3 was 3.1 ± 0.6 s. Distal aortic occlusion was not observed on either Day 1 or 7 in any group. Significantly more widespread and dense endothelial nitric oxide synthase (eNOS staining was observed in Group 1 animals than Group 2. On Days 1 and 7 after application of ABS, histopathological changes, consisting of necrosis, inflammation, and endothelial cell loss, in the rat abdominal aortas did not differ between Groups 1 and 2. The basophilic discoloration in the ABS group on the operation day was a result of a foreign body reaction and hemosiderin-loaded histiocyte accumulation, which occurred on Day 7. Conclusions In this study, hemostasis was successfully achieved with ABS in rat abdominal aortas. No histopathological change was found in the rat abdominal aortas between the ABS and control groups on Days 1 and 7. Further studies on the long-term effects of foreign body reactions and hemosiderin-loaded histiocyte

  15. Research of electrosurgical ablation with antiadhesive functionalization on thermal and histopathological effects of brain tissues in vivo.

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    Hsiao, Wen-Tien; Kung, Chun-Ming; Chu, Jan-Show; Ou, Keng-Liang; Peng, Pei-Wen

    2014-01-01

    Thermal injury and tissue sticking are two major concerns in the electrosurgery. In the present study, the effect of lateral thermal injury caused by different electrosurgical electrodes on wound healing was investigated. An electrosurgical unit equipped with untreated (SS) and titanium oxide layer-coated (TiO2-coated) stainless steel needle-type electrodes was used to create lesions on the rat brain tissue. TiO2 layers were produced by radiofrequency plasma and magnetron sputtering in the form of amorphous (TO-SS-1), anatase (TO-SS-2), and rutile (TO-SS-3) phase. Animals were sacrificed for evaluations at 0, 2, 7, and 28 days postoperatively. TO-SS-3 electrodes generated lower levels of sticking tissue, and the thermographs showed that the recorded highest temperature in brain tissue from the TO-SS-3 electrode was significantly lower than in the SS electrode. The total injury area of brain tissue caused by TO-SS-1 and TO-SS-3 electrodes was significantly lower than that caused by SS electrodes at each time point. The results of the present study reveal that the plating of electrodes with a TiO2 film with rutile phases is an efficient method for improving the performance of electrosurgical units and should benefit wound healing.

  16. Research of Electrosurgical Ablation with Antiadhesive Functionalization on Thermal and Histopathological Effects of Brain Tissues In Vivo

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    Wen-Tien Hsiao

    2014-01-01

    Full Text Available Thermal injury and tissue sticking are two major concerns in the electrosurgery. In the present study, the effect of lateral thermal injury caused by different electrosurgical electrodes on wound healing was investigated. An electrosurgical unit equipped with untreated (SS and titanium oxide layer-coated (TiO2-coated stainless steel needle-type electrodes was used to create lesions on the rat brain tissue. TiO2 layers were produced by radiofrequency plasma and magnetron sputtering in the form of amorphous (TO-SS-1, anatase (TO-SS-2, and rutile (TO-SS-3 phase. Animals were sacrificed for evaluations at 0, 2, 7, and 28 days postoperatively. TO-SS-3 electrodes generated lower levels of sticking tissue, and the thermographs showed that the recorded highest temperature in brain tissue from the TO-SS-3 electrode was significantly lower than in the SS electrode. The total injury area of brain tissue caused by TO-SS-1 and TO-SS-3 electrodes was significantly lower than that caused by SS electrodes at each time point. The results of the present study reveal that the plating of electrodes with a TiO2 film with rutile phases is an efficient method for improving the performance of electrosurgical units and should benefit wound healing.

  17. Postoperative morbidity and histopathologic characteristics of tonsillar tissue following coblation tonsillectomy in children: a prospective randomized single-blind study.

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    Roje, Zeljka; Racić, Goran; Dogas, Zoran; Pisac, Valdi Pesutić; Timms, Michael

    2009-03-01

    The aim of this prospective randomized single blind study was to determine the depth of thermal damage to tonsillar tissue due to coblation, and to compare it with thermal damage to tonsillar tissue following conventional tonsillectomy; to correlate the depth of thermal damage to tonsillar tissue with the parameters of postoperative morbidity, to compare intraoperative blood loss, postoperative pain severity, time to resuming normal physical activity, and incidence of postoperative bleeding between two groups of tonsillectomized children aged up to 16 years. 72 children aged 3-16 years scheduled for tonsillectomy randomly assigned into two groups submitted either to conventional tonsillectomy with bipolar diathermy coagulation or to coblation tonsillectomy, with a 14-day follow up. Statistically significant differences were observed in the depth of thermal damage to tonsillar tissue (p < 0.001), intraoperative blood loss (p < 0.004), in postoperative pain severity (p < 0.05) and in time to resuming normal physical activity between the two groups (p < 0.001). There was no case of reactionary or secondary bleeding in either group. In this paper for the first time we have correlated postoperative morbidity and thermal tissue damage: less thermal damage is associated with less postoperative morbidity.

  18. Optimizing Endoscopic Ultrasound Guided Tissue Sampling of the Pancreas

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    Pujan Kandel

    2016-03-01

    Full Text Available Endoscopic ultrasound is an important innovation in the field of gastrointestinal endoscopy and allows evaluation of many organs in the vicinity of the gastrointestinal tract. Endoscopic ultrasound-fine needle aspiration has been established to be an important tool in the management of pancreaticobiliary disease and is used for screening, staging, biopsy confirmation, and palliation. The accuracy of endoscopic ultrasound-fine needle aspiration is affected by several factors such as different needle sizes and types and fine needle aspiration techniques. Several comparative studies have been published on various techniques, such as the use of a stylet and suction during fine needle aspiration. Although most studies demonstrate high accuracy across techniques and equipment, various fine needle biopsy histology needles have been studied to compare the advantage of fine needle biopsy over fine needle aspiration. Although fine needle biopsy needles provide better tissue architecture and require fewer numbers of passes, there is no significant evidence of the superiority of fine needle biopsy over fine needle aspiration with regard to diagnostic yield and core tissue procurement. The main aim of this article is to review the various methodologies for improving the practice of endoscopic ultrasound-fine needle aspiration and endoscopic ultrasound- fine needle biopsy tissue sampling for cytological and histological analysis.

  19. Segmentation of colon tissue sample images using multiple graphics accelerators.

    Science.gov (United States)

    Szénási, Sándor

    2014-08-01

    Nowadays, processing medical images is increasingly done through using digital imagery and custom software solutions. The distributed algorithm presented in this paper is used to detect special tissue parts, the nuclei on haematoxylin and eosin stained colon tissue sample images. The main aim of this work is the development of a new data-parallel region growing algorithm that can be implemented even in an environment using multiple video accelerators. This new method has three levels of parallelism: (a) the parallel region growing itself, (b) starting more region growing in the device, and (c) using more than one accelerator. We use the split-and-merge technique based on our already existing data-parallel cell nuclei segmentation algorithm extended with a fast, backtracking-based, non-overlapping cell filter method. This extension does not cause significant degradation of the accuracy; the results are practically the same as those of the original sequential region growing method. However, as expected, using more devices usually means that less time is needed to process the tissue image; in the case of the configuration of one central processing unit and two graphics cards, the average speed-up is about 4-6×. The implemented algorithm has the additional advantage of efficiently processing very large images with high memory requirements. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. Histopathological changes in the liver and kidney tissues of marsh frog (Pelophylax ridibundus induced by the action of Talstar 10EC insecticide

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    Alina PĂUNESCU

    2012-05-01

    Full Text Available Histopathological biomarkers of toxicity in frog organs are a useful indicator of environmental pollution. The frogs were experimentally exposed to sub-lethal concentrations (0.5mg bifenthrin/g of body weight of Talstar 10EC for 3 weeks. The present study proves its toxic potential in terms of the damages in liver and kidney level. Tissues were normal in the control group. Hepatic lesions in frog exposed to bifenthrin were characterized by vacuolation of hepatocytes and nuclear pycnosis, perisinusoidal and periportal fibrosis, dilatation of Disse space and sinusoid capillaries, presence of cellular infiltrates. Kidney lesions consisted of dilation of Bowman’s space, tubular necrosis, and epithelial cells shows pycnotic nuclei, discrete degree of interstitial edema.

  1. Effect of rana galamensis–based diet on the activities of some enzymes and histopathology of selected tissues of albino rats

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    Basiru Olaitan Ajiboye

    2016-10-01

    Full Text Available The effect of Rana galamensis-based diet on the activities of some enzymes and histopathology of selected tissues of albino rats was investigated for eight weeks. A total of sixteen albino rats weighing between 29.15 and 26.01g (21 days old were divided into two groups. The first group contains animals fed on casein-based diet (control; the second group was fed on Rana galamensis-based diet. The animals were fed with their appropriate diet on daily basis and on the eight weeks of the experiment the animals were sacrificed using diethyl ether as anesthesia, blood was collected by cardiac puncture and organs of interest were harvested. Thereafter, organ to body weight ratio, some biochemical parameters and histopathology examination were carried out. There was no significant difference (p >0.05 in the organ to body weight ratio of the animals fed on control and Rana galamensis-based diets. Also, there was no significant different (p >0.05 in the activities of all the enzymes (ALP [alkaline phosphatase], AST [asparate transaminase], ALT [alanine transaminase], and γGT [gamma glutamyl transferase] investigated in the selected tissues and serum of rats fed on Rana galamensis- based diet when compared with the control. In addition, histological examinations of hepatocyte's rats fed on Rana galamensis- based diet show normal architecture structure when compared with the control. The insignificant different in the activities of all the enzymes studies (ALP, AST, ALT and γGT indicated no organ damage, supported by the normal histology studies. The obtained results may imply that Rana galamensis is safe for consumption.  Normal 0 false false false EN-US X-NONE X-NONE

  2. Reperfusion of the rat brain tissues following acute ischemia: the correlation among diffusion-weighted imaging, histopathology,and aquaporin-4 expression

    Institute of Scientific and Technical Information of China (English)

    LU Hong; HU Hui; HE Zhan-ping

    2011-01-01

    Background Although some studies have reported that aquaporin-4 (AQP4) plays a role in the post-ischemic edema formation and diffusion-weighted imaging (DWI), little is known about the AQP4 expression in stage of the reperfusion following acute cerebral ischemia, as well as the correlation between histopathology and DWl. The aim of the study was to investigate the correlation among DWl, histopathology and the AQP4 expression in the reperfused rat brain tissues following acute ischemia.Methods Seventy Wistar rats were randomly divided into a control group (group A), and several occluded and reperfusion groups. They had their middle cerebral artery unilaterally occluded (MCAO) for 30 minutes (group B) followed by 30 minutes (group D) or 60 minutes (group E) of reperfusion, or 60 minutes of MCAO (group C) followed by 30 minutes (group F), or 60 minutes (group G) of reperfusion (n=10 for each group). All rats underwent DWl scanning.The relative apparent diffusion coefficient (rADC) value of each rat was calculated. All the rats were sacrificed and the cerebral ischemic tissues were examined for histopathology. Real-time fluro-quantitative polymerase chain reaction (RT-PCR) and Western-blotting were performed. The amount of AQP4 mRNA (Ex △△Ct) and AQP4 protein (Q) was statistically analyzed. The correlation between rADC values and AQP4 mRNA expression was analyzed with the Pearson correlation test.Results In all the reperfusion groups, the areas of hyper-intensity signal in DWl were decreased, and the rADC value increased and the AQP4 expression decreased significantly compared with the occluded group (t=26.89, t=18.26, P<0.01). There was a negative correlation between AQP4 mRNA expression and rADC values (r=-0.72, P<0.01). A mixed edema, composed of cerebral intracelluar edema and vasogenic brain edema, was observed in all the reperfusion groups.It was more prevalent in groups D and F than in the groups E and G. With the reperfusion time postponed, the cerebral

  3. Anatomic, histopathologic, and echocardiographic features in a dog with an atypical pulmonary valve stenosis with a fibrous band of tissue and a patent ductus arteriosus.

    Science.gov (United States)

    Yoon, Hakyoung; Kim, Jaehwan; Nahm, Sang-Soep; Eom, Kidong

    2017-07-11

    Congenital pulmonary valve stenosis and patent ductus arteriosus are common congenital heart defects in dogs. However, concurrence of atypical pulmonary valve stenosis and patent ductus arteriosus is uncommon. This report describes the anatomic, histopathologic, and echocardiographic features in a dog with concomitant pulmonary valve stenosis and patent ductus arteriosus with atypical pulmonary valve dysplasia that included a fibrous band of tissue. A 1.5-year-old intact female Chihuahua dog weighing 3.3 kg presented with a continuous grade VI cardiac murmur, poor exercise tolerance, and an intermittent cough. Echocardiography indicated pulmonary valve stenosis, a thickened dysplastic valve without annular hypoplasia, and a type IIA patent ductus arteriosus. The pulmonary valve was thick line-shaped in systole and dome-shaped towards the right ventricular outflow tract in diastole. The dog suffered a fatal cardiac arrest during an attempted balloon pulmonary valvuloplasty. Necropsy revealed pulmonary valve dysplasia, commissural fusion, and incomplete opening and closing of the pulmonary valve because of a fibrous band of tissue causing adhesion between the right ventricular outflow tract and the dysplastic intermediate cusp of the valve. A fibrous band of tissue between the right ventricular outflow track and the pulmonary valve should be considered as a cause of pulmonary valve stenosis. Pulmonary valve stenosis and patent ductus arteriosus can have conflicting effects on diastolic and systolic dysfunction, respectively. Therefore, beta-blockers should always be used carefully, particularly in patients with a heart defect where there is concern about left ventricular systolic function.

  4. Histopathology for the diagnosis of infectious diseases

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    Gupta E

    2009-01-01

    Full Text Available Histopathological examination of tissue biopsies for the identification of infectious organisms is a very important diagnostic tool. Conventional culture confirmation of tissue biopsies often fail to identify any pathogen as, first of all, invariably most of the tissue samples that are collected and sent for culture isolation are inappropriately collected in formalin, which prevents pathogen growth in culture media. Inadequate processing like grinding, etc. further hinders isolation. Presence of inhibitors like dead tissue debris, fibers, etc. also delays isolation. Microbiologists often lack expertise in identifying infectious pathogens directly from tissue biopsies by microscopic visualization. This review therefore acquaints microbiologists with the various methods available for detecting infectious agents by using histological stains. On histopathological examination of the tissue biopsy once, it is determined that a disease is likely to be due to an infection and has characterized the inflammatory response and hence associated microorganisms should be thoroughly looked for. Although some microorganisms or their cytopathic effects may be clearly visible on routine haematoxylin- and eosin-stained sections, additional histochemical stains are often needed for their complete characterization. Highly specific molecular techniques, such as immunohistochemistry, in situ hybridization and nucleic acid amplification, may be needed in certain instances to establish the diagnosis of infection. Through appropriate morphologic diagnoses and interlaboratory communication and collaboration, direct microscopic visualization of tissue samples can thus be very helpful in reaching a correct and rapid diagnosis.

  5. Histopathological changes of renal tissue following sodium fluoride administration in two consecutive generations of mice. Correlation with the urinary elimination of fluoride.

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    Dimcevici Poesina, Nicoleta; Bălălău, Cristian; Nimigean, Vanda Roxana; Nimigean, Victor; Ion, Ion; Baconi, Daniela; Bârcă, Maria; Băran Poesina, Violeta

    2014-01-01

    The present study was designed to investigate the toxic effects (evaluated as histopathological changes) of sodium fluoride on the kidney in two consecutive generations of NMRI mice. An attempt to correlate the toxicity with the urinary elimination of fluoride has been made, as urinary fluoride excretion has been widely used as an indicator of fluoride intake and exposure. Six mixed (males and females) animal groups have been constituted by dividing the populations of mice derived from pregnant females (named "mothers" 0.5 mg sodium fluoride) treated with 0.5 mg sodium fluoride by daily gavage and pregnant females (named "mothers" 0.25 mg sodium fluoride) treated with 0.25 mg sodium fluoride by daily gavage; three types of sodium fluoride treatments were administrated: homeopathic, allopathic-homeopathic and allopathic. When the animals reached the adulthood, by randomization, they were selected in pairs for giving birth to the second generation of mice. No treatments were administrated to the second generation of mice; thus, the urinary elimination of fluoride in the second generation is attributed to exposure at sodium fluoride before birth. The administration of sodium fluoride to the first generation (F1) is realized until the mice reached the adulthood. For the first generation, the urine was collected at three times, every three weeks: at the age of four weeks, seven weeks and 11 weeks; single sampling urine, at the age of four weeks, has been conducted for the second generation. The urine samples have been analyzed using the ion selective electrode method for fluoride. For the histopathological examination, the animals were killed by cervical dislocation; the kidneys were collected in a 10% formalin solution. The preparation of samples for optical microscopy was realized with Hematoxylin-Eosin staining. The results indicate that the elimination of fluoride was similar (at the second evaluation, at 7-week-old of the first generation) for the both generations

  6. Investigation of biochemical and histopathological effects of Mentha piperita L. and Mentha spicata L. on kidney tissue in rats.

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    Akdogan, Mehmet; Kilinç, Ibrahim; Oncu, Meral; Karaoz, Erdal; Delibas, Namik

    2003-04-01

    Peppermint plants have been used as a herbal medicine for many conditions, including loss of appetite, common cold, bronchitis, sinusitis, fever, nausea, vomiting and indigestion. This study is aimed at investigating the biochemical and histological effects of Mentha piperita L., growing in the Yenisar Bademli town of Isparta City, and Mentha spicata L., growing on the Anamas high plateau of Isparta City, on rat kidney tissue. Forty-eight male Wistar albino rats weighing 200-250 g were used for this study. Animals were divided into four experimental groups, each with 12 rats, as follows: control group (group I); 20 g/L M. piperita tea (group II); 20 g/L M. spicata tea (group III); 40 g/L M. spicata tea (group IV). The control group rats were given commercial drinking water (Hayat DANONESA water). The tea for the other groups was prepared daily and provided at all times to the rats during 30 days as drinking water. Plasma urea and creatinine levels were determined, and the levels of thiobarbituric acid reactive substance (TBARS) and the activities of glutathione peroxidase (GSH-Px), catalase (CAT) and superoxide dismutase (SOD) were studied in the homogenates of kidney tissue. The levels of plasma urea and creatinine were increased significantly (P spicata presents markedly nephrotoxic changes in rats.

  7. The histopathological effects of superovulatory drugs GnRHα/PMSG/HCG on the endometrial tissues of rat

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    Parviz Bazzi

    2005-09-01

    Full Text Available Background: Using of stimulatory drugs such as: GnRHα/ PMSG/ HCG for induction of superovulation and harvesting more oocytes is very common during Assisted Reproductive Techniques (ART. Studies and experiences have recently shown that the rate of successful implantation in stimulatory cycles is less than natural cycles. Methods: The possible effects of above mentioned drugs on ultrastructures of endometrial tissues (Luminal epithelium was done by endometrial biopsies from female rats (N=30 which were under superovulaion treatment and from control rats with normal menstrual cycles (N=30. The specimens were processed for electronmicroscopic studies. Qualitative and quantitative (morphologic and morphometric studies were carried out on electronmicrographs. Morphological findings based on three structures namely: nuclear channel system (NCS, giant mitochondria (GM and glycogen vacuoles in addition to pinopode system and nuclear euchromatin were evaluated. Results: In morphology, luminal epithelium was more active and had more secretions in rats under superovulation compared with control group. Morphometric findings showed that in case group, volume fraction of nucleus RER, mitochondrium, glycogen to cell and euchromatin to nucleus had statistically significant difference. Conclusion: Superovulatory drugs GnRHα/PMSG/HCG have a negative effect on endometrial tissues at the time of implantation which may lead to low implantation rate.

  8. CYTOKINE-PRODUCING RESERVE OF IMMUNOCOMPETENT CELLS FROM BLOOD AND INVASIVE DUCTAL BREAST CANCER TISSUES: ITS CORRELATION WITH HISTOPATHOLOGICAL AND IMMUNOHISTOCHEMICAL PARAMETERS OF MALIGNANT NEOPLASIA

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    S. A. Arkhipov

    2016-01-01

    Full Text Available The aim of the study was to investigate a relationship between cytokine-producing reserve of invasive ductal cancer cells and its microenvironment, and cytokine-producing reserve of immunocompetent blood cells (IBC, as well as with histopathological and immunohistochemical characteristics of breast cancer. Using ELISA method we investigated the spontaneous and stimulated with polyclonal activators (PA cytokine-producing reserve of IBC and biopsy specimens from invasive ductal cancer (adenocarcinoma in 34 women. Appropriate values were expressed by the Influence Index of polyclonal activators (IIPA upon cytokine production (IL-2, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β, IL-1ra, TNFα, IFNγ, G-CSF, GM-CSF, VEGF-A, MCP-1. In tumor biopsies, we studied expression of VEGF-A, estrogen receptor, progesteron receptor and pro-proliferation marker Ki-67 by means of immunohistochemical method. Activation values of blood IBC in most cases, except of IL-18, IL-1β and MCP-1, were higher than appropriate effects upon cytokine production by tumor tisuues. Meanwhile, the IIPA activation index upon IL-18 (a proinflammatory and prooncogene cytokine production by tumor cells and its microenvironment proved to be elevated, as compared to appropriate IIPA by the blood IBC. Statistical studies showed a direct correlation between IIPA and cytokine production in tumor supernates, IIPA of VEGF-A expression in tumor tissue, with pathohistological parameters and expression of estrogen and progesterone receptors and Ki-67 proliferation marker. A high positive correlation was obtained between IIPA TNFα production by the tumor tissue, and degree of tumor vascularization.We have revealed a negative correlation between IIPA for IL-6, MCP-1 and Ki-67 marker of cell proliferation. A direct correlation was found between IIPA values for IL-1ra/IL-1β production ratios in blood cells, and IIPA for VEGF-A expression in adenocarcinoma tissues, thus indicating to probable

  9. Artefacts in histopathology.

    Science.gov (United States)

    Chatterjee, Shailja

    2014-09-01

    Histopathology is the science of slide analysis for the diagnostic and research purposes. However, sometimes the presence of certain artefacts in a microscopic section can result in misinterpretations leading to diagnostic pitfalls that can result in increased patient morbidity. This article reviews the common artefacts encountered during slide examination alongside the remedial measures which can be undertaken to differentiate between an artefact and tissue constituent.

  10. Artefacts in histopathology

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    Shailja Chatterjee

    2014-01-01

    Full Text Available Histopathology is the science of slide analysis for the diagnostic and research purposes. However, sometimes the presence of certain artefacts in a microscopic section can result in misinterpretations leading to diagnostic pitfalls that can result in increased patient morbidity. This article reviews the common artefacts encountered during slide examination alongside the remedial measures which can be undertaken to differentiate between an artefact and tissue constituent.

  11. Nail histopathology.

    Science.gov (United States)

    Martin, B

    2013-09-01

    The structure of the nail unit is complex and many dermatologists and dermatopathologists have an incomplete understanding of it. Familiarity with the anatomy and histology of this unit, however, is a key factor in improving the diagnostic yield of nail biopsy. Inflammatory or infectious conditions that affect the nail can have a marked impact on a patient's quality of life. A wide-ranging variety of tumors can also develop in this region and they may be life-threatening or require surgery that will result in functional defects. The author reviews the anatomy and histology of the nail unit as well as the basic histopathologic findings in the most common conditions affecting the nails. Copyright © 2011 Elsevier España, S.L. and AEDV. All rights reserved.

  12. Biochemical imaging of normal, adenoma, and colorectal adenocarcinoma tissues by Fourier transform infrared spectroscopy (FTIR and morphological correlation by histopathological analysis: preliminary results

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    Juliana Aparecida de Almeida Chaves Piva

    Full Text Available Introduction The colorectal cancer is a major health problem worldwide. Histology is considered the gold standard for differential diagnosis. However, it depends on the observer's experience, which can lead to discrepancies and poor results. Spectroscopic imaging by Fourier transform infrared (FTIR is a technique that may be able to improve the diagnosis, because it is based on biochemical differences of the structural constituents of tissue. Therefore, the main goal of this study was to explore the use of FTIR imaging technique in normal colon tissue, colorectal adenoma, and adenocarcinoma in order to correlate their morphological structures with their biochemical imaging. Methods Samples were collected from normal (n = 4, adenoma (n = 4, and adenocarcinoma human colorectal tissue (n = 4 from patients undergoing colonoscopy or surgical resection of colon lesions. The samples were sectioned with a cryostat in sequential sections; the first slice was placed on CaF2 slide and the second slice was placed on glass slide for histological analysis (HE staining. The cluster analyses were performed by the software Cytospec (1.4.02®. Results In normal samples, biochemical analysis classified six different structures, namely the lamina propria of mucous glands (epithelial cells and goblet cells, central lumen of the gland, mucin, and conjunctive tissue. In samples with adenoma and adenocarcinoma, altered regions could also be identified with high sensitivity and specificity. Conclusion The results of this study demonstrate the potential and viability of using infrared spectroscopy to identify and classify colorectal tissues.

  13. Long-term room temperature preservation of corpse soft tissue: an approach for tissue sample storage

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    Caputo Mariela

    2011-08-01

    Full Text Available Abstract Background Disaster victim identification (DVI represents one of the most difficult challenges in forensic sciences, and subsequent DNA typing is essential. Collected samples for DNA-based human identification are usually stored at low temperature to halt the degradation processes of human remains. We have developed a simple and reliable procedure for soft tissue storage and preservation for DNA extraction. It ensures high quality DNA suitable for PCR-based DNA typing after at least 1 year of room temperature storage. Methods Fragments of human psoas muscle were exposed to three different environmental conditions for diverse time periods at room temperature. Storage conditions included: (a a preserving medium consisting of solid sodium chloride (salt, (b no additional substances and (c garden soil. DNA was extracted with proteinase K/SDS followed by organic solvent treatment and concentration by centrifugal filter devices. Quantification was carried out by real-time PCR using commercial kits. Short tandem repeat (STR typing profiles were analysed with 'expert software'. Results DNA quantities recovered from samples stored in salt were similar up to the complete storage time and underscored the effectiveness of the preservation method. It was possible to reliably and accurately type different genetic systems including autosomal STRs and mitochondrial and Y-chromosome haplogroups. Autosomal STR typing quality was evaluated by expert software, denoting high quality profiles from DNA samples obtained from corpse tissue stored in salt for up to 365 days. Conclusions The procedure proposed herein is a cost efficient alternative for storage of human remains in challenging environmental areas, such as mass disaster locations, mass graves and exhumations. This technique should be considered as an additional method for sample storage when preservation of DNA integrity is required for PCR-based DNA typing.

  14. Quantitation of ranaviruses in cell culture and tissue samples.

    Science.gov (United States)

    Holopainen, Riikka; Honkanen, Jarno; Jensen, Britt Bang; Ariel, Ellen; Tapiovaara, Hannele

    2011-01-01

    A quantitative real-time PCR (qPCR) based on a standard curve was developed for detection and quantitation of ranaviruses. The target gene for the qPCR was viral DNA polymerase (DNApol). All ten ranavirus isolates studied (Epizootic haematopoietic necrosis virus, EHNV; European catfish virus, ECV; European sheatfish virus, ESV; Frog virus 3, FV3; Bohle iridovirus, BIV; Doctor fish virus, DFV; Guppy virus 6, GV6; Pike-perch iridovirus, PPIV; Rana esculenta virus Italy 282/I02, REV282/I02 and Short-finned eel ranavirus, SERV) were detected with the qPCR assay. In addition, two fish cell lines - epithelioma papulosum cyprini (EPC) and bluegill fry (BF-2) - were infected with four of the isolates (EHNV, ECV, FV3 and DFV), and the viral quantity was determined from seven time points during the first three days after infection. The qPCR was also used to determine the viral load in tissue samples from pike (Esox lucius) fry challenged experimentally with EHNV. Copyright © 2010 Elsevier B.V. All rights reserved.

  15. Mapping differential elemental accumulation in fish tissues: Importance of fish tissue sampling standardization

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    Jovičić Katarina

    2016-01-01

    Full Text Available The concentrations of As, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Se and Zn in the muscle, gills, liver and intestine of the wels catfish (Silurus glanis from the Danube River were analyzed by inductively coupled plasma mass spectrometry (ICP-MS. The aim of the study was to determine whether in complex muscle/skin, gill filament/gill arch, proximal/distal liver and proximal/median/distal intestine samples, particular components differ in concentrations of the analyzed elements. Results indicated that there were no differences in the accumulation of different elements between the proximal and distal liver segments and between the proximal and median intestine sections. Conversely, elemental accumulation patterns in muscle and skin differed significantly. Significant differences were also observed between the gill arch and filaments, as well as between the distal and the two upper intestine sections. Findings indicated the importance of detailed reporting of tissue sampling, i.e. whether the skin was included in the muscle sample, as well as if the gill arch and filaments were analyzed together. Due to a potential bias that can be produced by different muscle/skin or gill arch/filament ratios included in the sample, we strongly recommend that they should not be analyzed together. Results of the present study might be of interest to the scientific community and stakeholders involved in aquatic ecosystem monitoring programs. [Projekat Ministarstva nauke Republike Srbije, br. TR37009 i br. 173045

  16. Versatile electrochemial sensor for tissue culturing and sample handling

    DEFF Research Database (Denmark)

    Bakmand, Tanya; Kwasny, Dorota; Al Atraktchi, Fatima Al-Zahraa

    2014-01-01

    Culturing of organtypic brain tissues is a routine procedure in neural research. The visual inspection of the medium is the only way of determining the state of the tissue. At the end of culturing, post-processing techniques such as HPLC can be used to measure the concentration of the secreted me...

  17. Histopathology and bioaccumulation of heavy metals (Cr, Ni and Pb) in fish (Channa striatus and Heteropneustes fossilis) tissue: a study for toxicity and ecological impacts.

    Science.gov (United States)

    Fatima, Mahino; Usmani, Nazura

    2013-05-01

    Abstract: The water samples were collected from the 22 km segment III of Yamuna River from Okhla barrage. This segment receives water from 17 sewage drains of Delhi, Western Yamuna Canal (WYC), upper Ganga canal via Najafgarh drain and Hindon cut canal. Hence, the water samples collected were used to determine the presence of Chromium, Nickel and Lead through Atomic Absorption Spectrophotometry. The concentration of these heavy metals were much above the maximum permissible limits set by WHO. This was bound to have its influence on the riverine flora and fauna. To evaluate this, two popularly consumed fish species such as Channa striatus and Heteropneustes fossilis were caught and the bioaccumulation of these heavy metals were estimated in different organs (liver, kidney, gill and muscle). It was found that Cr accumulated the most in these organs (gill being most influenced) in both the species. The accumulation of all these heavy metals were above MPL set by World Health Organisation (WHO) and Food and Drug Administration (FDA). Histopathology was also conducted where heavy damages were observed in both liver and kidney of both the species.

  18. Variation in glycogen concentrations within mantle and foot tissue in Amblema plicata plicata: Implications for tissue biopsy sampling

    Science.gov (United States)

    Naimo, T.J.; Monroe, E.M.

    1999-01-01

    With the development of techniques to non-lethally biopsy tissue from unionids, a new method is available to measure changes in biochemical, contaminant, and genetic constituents in this imperiled faunal group. However, before its widespread application, information on the variability of biochemical components within and among tissues needs to be evaluated. We measured glycogen concentrations in foot and mantle tissue in Amblema plicata plicata (Say, 1817) to determine if glycogen was evenly distributed within and between tissues and to determine which tissue might be more responsive to the stress associated with relocating mussels. Glycogen was measured in two groups of mussels: those sampled from their native environment (undisturbed mussels) and quickly frozen for analysis and those relocated into an artificial pond (relocated mussels) for 24 months before analysis. In both undisturbed and relocated mussels, glycogen concentrations were evenly distributed within foot, but not within mantle tissue. In mantle tissue, concentrations of glycogen varied about 2-fold among sections. In addition, glycogen varied significantly between tissues in undisturbed mussels, but not in relocated mussels. Twenty-four months after relocation, glycogen concentrations had declined by 80% in mantle tissue and by 56% in foot tissue relative to the undisturbed mussels. These data indicate that representative biopsy samples can be obtained from foot tissue, but not mantle tissue. We hypothesize that mantle tissue could be more responsive to the stress of relocation due to its high metabolic activity associated with shell formation.

  19. Evidence of presence of Mycobacterium tuberculosis in bovine tissue samples by multiplex PCR: possible relevance to reverse zoonosis.

    Science.gov (United States)

    Mittal, M; Chakravarti, S; Sharma, V; Sanjeeth, B S; Churamani, C P; Kanwar, N S

    2014-04-01

    Bovine tuberculosis, caused by Mycobacterium bovis, remains one of the most important zoonotic health concerns worldwide. The transmission of Mycobacterium tuberculosis from humans to animals also occurs especially in countries where there is close interaction of humans with the animals. In the present study, thirty bovine lung tissue autopsy samples from an organized dairy farm located in North India were screened for the presence of Mycobacterium tuberculosis complex by smear microscopy, histopathological findings and PCR. Differential diagnosis of M. tuberculosis and M. bovis was made based on the deletion of mce-3 operon in M. bovis. The present study found eight of these samples positive for M. tuberculosis by multiplex PCR. Sequencing was performed on two PCR-positive representative samples and on annotation, and BLAST analysis confirmed the presence of gene fragment specific to Mycobacterium tuberculosis. The presence of M. tuberculosis in all the positive samples raises the possibility of human-to-cattle transmission and possible adaptation of this organism in bovine tissues. This study accentuates the importance of screening and differential diagnosis of Mycobacterium tuberculosis complex in humans and livestock for adopting effective TB control and eradication programmes.

  20. Histopathological changes in tendinopathy--potential roles of BMPs?

    Science.gov (United States)

    Lui, Pauline Po Yee

    2013-12-01

    The pathogenesis of tendinopathy remains unclear. Chondro-osteogenic BMPs such as BMP-2, BMP-4 and BMP-7 have been reported in clinical samples and animal models of tendinopathy. As chondrocyte-like cells and ossified deposits have been observed in both clinical samples and animal models of failed tendon healing tendinopathy, chondro-osteogenic BMPs might contribute to tissue metaplasia and other histopathological changes in tendinopathy. In this review I have summarized the current evidence supporting the roles of chondro-osteogenic BMPs in the histopathological changes of tendinopathy. The potential targets, effects and sources of these BMPs are discussed. I have also provided directions for future studies about the potential roles of BMPs in the pathogenesis of tendinopathy. Better understanding of the roles of these BMPs in the histopathological changes of tendinopathy could provide new options for the prevention and treatment of this disabling tendon disorder.

  1. Out-of-Sample Extrapolation utilizing Semi-Supervised Manifold Learning (OSE-SSL): Content Based Image Retrieval for Histopathology Images.

    Science.gov (United States)

    Sparks, Rachel; Madabhushi, Anant

    2016-06-06

    Content-based image retrieval (CBIR) retrieves database images most similar to the query image by (1) extracting quantitative image descriptors and (2) calculating similarity between database and query image descriptors. Recently, manifold learning (ML) has been used to perform CBIR in a low dimensional representation of the high dimensional image descriptor space to avoid the curse of dimensionality. ML schemes are computationally expensive, requiring an eigenvalue decomposition (EVD) for every new query image to learn its low dimensional representation. We present out-of-sample extrapolation utilizing semi-supervised ML (OSE-SSL) to learn the low dimensional representation without recomputing the EVD for each query image. OSE-SSL incorporates semantic information, partial class label, into a ML scheme such that the low dimensional representation co-localizes semantically similar images. In the context of prostate histopathology, gland morphology is an integral component of the Gleason score which enables discrimination between prostate cancer aggressiveness. Images are represented by shape features extracted from the prostate gland. CBIR with OSE-SSL for prostate histology obtained from 58 patient studies, yielded an area under the precision recall curve (AUPRC) of 0.53 ± 0.03 comparatively a CBIR with Principal Component Analysis (PCA) to learn a low dimensional space yielded an AUPRC of 0.44 ± 0.01.

  2. IHC Profiler: an open source plugin for the quantitative evaluation and automated scoring of immunohistochemistry images of human tissue samples.

    Directory of Open Access Journals (Sweden)

    Frency Varghese

    Full Text Available In anatomic pathology, immunohistochemistry (IHC serves as a diagnostic and prognostic method for identification of disease markers in tissue samples that directly influences classification and grading the disease, influencing patient management. However, till today over most of the world, pathological analysis of tissue samples remained a time-consuming and subjective procedure, wherein the intensity of antibody staining is manually judged and thus scoring decision is directly influenced by visual bias. This instigated us to design a simple method of automated digital IHC image analysis algorithm for an unbiased, quantitative assessment of antibody staining intensity in tissue sections. As a first step, we adopted the spectral deconvolution method of DAB/hematoxylin color spectra by using optimized optical density vectors of the color deconvolution plugin for proper separation of the DAB color spectra. Then the DAB stained image is displayed in a new window wherein it undergoes pixel-by-pixel analysis, and displays the full profile along with its scoring decision. Based on the mathematical formula conceptualized, the algorithm is thoroughly tested by analyzing scores assigned to thousands (n = 1703 of DAB stained IHC images including sample images taken from human protein atlas web resource. The IHC Profiler plugin developed is compatible with the open resource digital image analysis software, ImageJ, which creates a pixel-by-pixel analysis profile of a digital IHC image and further assigns a score in a four tier system. A comparison study between manual pathological analysis and IHC Profiler resolved in a match of 88.6% (P<0.0001, CI = 95%. This new tool developed for clinical histopathological sample analysis can be adopted globally for scoring most protein targets where the marker protein expression is of cytoplasmic and/or nuclear type. We foresee that this method will minimize the problem of inter-observer variations across labs and

  3. IHC Profiler: An Open Source Plugin for the Quantitative Evaluation and Automated Scoring of Immunohistochemistry Images of Human Tissue Samples

    Science.gov (United States)

    Malhotra, Renu; De, Abhijit

    2014-01-01

    In anatomic pathology, immunohistochemistry (IHC) serves as a diagnostic and prognostic method for identification of disease markers in tissue samples that directly influences classification and grading the disease, influencing patient management. However, till today over most of the world, pathological analysis of tissue samples remained a time-consuming and subjective procedure, wherein the intensity of antibody staining is manually judged and thus scoring decision is directly influenced by visual bias. This instigated us to design a simple method of automated digital IHC image analysis algorithm for an unbiased, quantitative assessment of antibody staining intensity in tissue sections. As a first step, we adopted the spectral deconvolution method of DAB/hematoxylin color spectra by using optimized optical density vectors of the color deconvolution plugin for proper separation of the DAB color spectra. Then the DAB stained image is displayed in a new window wherein it undergoes pixel-by-pixel analysis, and displays the full profile along with its scoring decision. Based on the mathematical formula conceptualized, the algorithm is thoroughly tested by analyzing scores assigned to thousands (n = 1703) of DAB stained IHC images including sample images taken from human protein atlas web resource. The IHC Profiler plugin developed is compatible with the open resource digital image analysis software, ImageJ, which creates a pixel-by-pixel analysis profile of a digital IHC image and further assigns a score in a four tier system. A comparison study between manual pathological analysis and IHC Profiler resolved in a match of 88.6% (P<0.0001, CI = 95%). This new tool developed for clinical histopathological sample analysis can be adopted globally for scoring most protein targets where the marker protein expression is of cytoplasmic and/or nuclear type. We foresee that this method will minimize the problem of inter-observer variations across labs and further help in

  4. Histopathological characteristics of human non-tumor thyroid tissues in a long-term model of adenomatous goiter xenografts in the NOD/Shi-scid, IL-2Rγ(null) mouse.

    Science.gov (United States)

    Fujii, Etsuko; Kato, Atsuhiko; Chen, Yu Jau; Matsubara, Koichi; Ohnishi, Yasuyuki; Suzuki, Masami

    2014-07-01

    There is a growing need for modeling the human thyroid to link data obtained from animals to humans because of its sensitivity to radiation exposure and endocrine disruption chemicals. In a scid mouse model produced by transplanting human thyroid tissues, leakiness and thymic lymphoma that occurs spontaneously in the scid mouse can complicate the interpretation of experimental results. Considering that the NOD.Cg-Prkdc(scid)Il2rg(tm1Sug)/Jic mouse (NOD/Shi-scid, IL-2Rγ(null) or NOG mouse) may be a better host because this strain has low incidence of leakiness and thymic lymphoma, we have evaluated the potential of a model that allows long-term observation of non-tumor human thyroid tissues in this mouse. We transplanted tissues of human adenomatous goiter into NOG mice and examined the tissues histopathologically. The morphology of human adenomatous goiter tissues was maintained from 24 to 44 weeks after transplantation in NOG mice with no noted differences between donor-matched tissues or the weeks after transplantation. The tissues expressed thyroglobulin protein and mRNA as well as thyroperoxidase. Endothelial cells originating from human were found in the transplanted tissues and were thought to be a characteristic of this model. The intactness of the tissues before transplantation was found to affect the rate of tissue engraftment. From the present results we have concluded that transplanted thyroid tissues in NOG mice maintain the histopathological characteristics of their origin for long terms. Therefore this model was thought feasible for toxicity evaluation. Copyright © 2014 Elsevier GmbH. All rights reserved.

  5. Final LDRD report : development of sample preparation methods for ChIPMA-based imaging mass spectrometry of tissue samples.

    Energy Technology Data Exchange (ETDEWEB)

    Maharrey, Sean P.; Highley, Aaron M.; Behrens, Richard, Jr.; Wiese-Smith, Deneille

    2007-12-01

    The objective of this short-term LDRD project was to acquire the tools needed to use our chemical imaging precision mass analyzer (ChIPMA) instrument to analyze tissue samples. This effort was an outgrowth of discussions with oncologists on the need to find the cellular origin of signals in mass spectra of serum samples, which provide biomarkers for ovarian cancer. The ultimate goal would be to collect chemical images of biopsy samples allowing the chemical images of diseased and nondiseased sections of a sample to be compared. The equipment needed to prepare tissue samples have been acquired and built. This equipment includes an cyro-ultramicrotome for preparing thin sections of samples and a coating unit. The coating unit uses an electrospray system to deposit small droplets of a UV-photo absorbing compound on the surface of the tissue samples. Both units are operational. The tissue sample must be coated with the organic compound to enable matrix assisted laser desorption/ionization (MALDI) and matrix enhanced secondary ion mass spectrometry (ME-SIMS) measurements with the ChIPMA instrument Initial plans to test the sample preparation using human tissue samples required development of administrative procedures beyond the scope of this LDRD. Hence, it was decided to make two types of measurements: (1) Testing the spatial resolution of ME-SIMS by preparing a substrate coated with a mixture of an organic matrix and a bio standard and etching a defined pattern in the coating using a liquid metal ion beam, and (2) preparing and imaging C. elegans worms. Difficulties arose in sectioning the C. elegans for analysis and funds and time to overcome these difficulties were not available in this project. The facilities are now available for preparing biological samples for analysis with the ChIPMA instrument. Some further investment of time and resources in sample preparation should make this a useful tool for chemical imaging applications.

  6. Micro-organisms isolated from cadaveric samples of allograft musculoskeletal tissue.

    Science.gov (United States)

    Varettas, Kerry

    2013-12-01

    Allograft musculoskeletal tissue is commonly used in orthopaedic surgical procedures. Cadaveric donors of musculoskeletal tissue supply multiple allografts such as tendons, ligaments and bone. The microbiology laboratory of the South Eastern Area Laboratory Services (SEALS, Australia) has cultured cadaveric allograft musculoskeletal tissue samples for bacterial and fungal isolates since 2006. This study will retrospectively review the micro-organisms isolated over a 6-year period, 2006-2011. Swab and tissue samples were received for bioburden testing and were inoculated onto agar and/or broth culture media. Growth was obtained from 25.1 % of cadaveric allograft musculoskeletal tissue samples received. The predominant organisms isolated were coagulase-negative staphylococci and coliforms, with the heaviest bioburden recovered from the hemipelvis. The rate of bacterial and fungal isolates from cadaveric allograft musculoskeletal tissue samples is higher than that from living donors. The type of organism isolated may influence the suitability of the allograft for transplant.

  7. Histopathological placental lesions in mild gestational hyperglycemic and diabetic women

    Directory of Open Access Journals (Sweden)

    Rudge Marilza VC

    2011-08-01

    Full Text Available Abstract Objective To investigate and compare the incidence of histopathological placental lesions in mild gestational hyperglycemia, gestational diabetes and overt diabetes at term and preterm gestation. Research design and methods One-hundred-and-thirty-one placental samples were collected from Diabetes mellitus (DM positive screened patients. Two diagnostic tests, glycemic profile and 100 g oral glucose tolerance test (OGTT in parallel identified 4 groups normoglycemic, mild gestational hyperglycemia (MGH, gestational DM (GDM or overt DM (DM. Placental tissue specimens and sections from 4 groups were obtained by uniform random sampling and stained with hematoxylin-eosin. Results Placentas from MGH group presented 17 types of histopathological change and higher rates of syncytial nodes and endarteritis. GDM placentas presented only nine types of histopathological change, high rates of dysmaturity, low rates of calcification and no syncytial nodes. Overt DM placentas showed 22 types of histopathological change, 21 of which were present in the preterm period. There were histopathological similarities between MGH and DM placentas, but the former exhibited a higher incidence of endarteritis, which has been described as a "post-mortem" phenomenon. Conclusion Our results confirmed that the distinct placental changes associated with DM and MGH depend on gestational period during which the diabetic insult occurs. It may reasonably be inferred that subclinical maternal hyperglycemia during pregnancy, as showed in MGH group, is responsible for increased placental endarteritis, a postmortem lesion in the live fetus.

  8. Elemental distribution and sample integrity comparison of freeze-dried and frozen-hydrated biological tissue samples with nuclear microprobe

    Energy Technology Data Exchange (ETDEWEB)

    Vavpetič, P., E-mail: primoz.vavpetic@ijs.si [Jožef Stefan Institute, Jamova 39, SI-1000 Ljubljana (Slovenia); Vogel-Mikuš, K. [Biotechnical Faculty, Department of Biology, University of Ljubljana, Jamnikarjeva 101, SI-1000 Ljubljana (Slovenia); Jeromel, L. [Jožef Stefan Institute, Jamova 39, SI-1000 Ljubljana (Slovenia); Ogrinc Potočnik, N. [Jožef Stefan Institute, Jamova 39, SI-1000 Ljubljana (Slovenia); FOM-Institute AMOLF, Science Park 104, 1098 XG Amsterdam (Netherlands); Pongrac, P. [Biotechnical Faculty, Department of Biology, University of Ljubljana, Jamnikarjeva 101, SI-1000 Ljubljana (Slovenia); Department of Plant Physiology, University of Bayreuth, Universitätstr. 30, 95447 Bayreuth (Germany); Drobne, D.; Pipan Tkalec, Ž.; Novak, S.; Kos, M.; Koren, Š.; Regvar, M. [Biotechnical Faculty, Department of Biology, University of Ljubljana, Jamnikarjeva 101, SI-1000 Ljubljana (Slovenia); Pelicon, P. [Jožef Stefan Institute, Jamova 39, SI-1000 Ljubljana (Slovenia)

    2015-04-01

    The analysis of biological samples in frozen-hydrated state with micro-PIXE technique at Jožef Stefan Institute (JSI) nuclear microprobe has matured to a point that enables us to measure and examine frozen tissue samples routinely as a standard research method. Cryotome-cut slice of frozen-hydrated biological sample is mounted between two thin foils and positioned on the sample holder. The temperature of the cold stage in the measuring chamber is kept below 130 K throughout the insertion of the samples and the proton beam exposure. Matrix composition of frozen-hydrated tissue is consisted mostly of ice. Sample deterioration during proton beam exposure is monitored during the experiment, as both Elastic Backscattering Spectrometry (EBS) and Scanning Transmission Ion Microscopy (STIM) in on–off axis geometry are recorded together with the events in two PIXE detectors and backscattered ions from the chopper in a single list-mode file. The aim of this experiment was to determine differences and similarities between two kinds of biological sample preparation techniques for micro-PIXE analysis, namely freeze-drying and frozen-hydrated sample preparation in order to evaluate the improvements in the elemental localisation of the latter technique if any. In the presented work, a standard micro-PIXE configuration for tissue mapping at JSI was used with five detection systems operating in parallel, with proton beam cross section of 1.0 × 1.0 μm{sup 2} and a beam current of 100 pA. The comparison of the resulting elemental distributions measured at the biological tissue prepared in the frozen-hydrated and in the freeze-dried state revealed differences in elemental distribution of particular elements at the cellular level due to the morphology alteration in particular tissue compartments induced either by water removal in the lyophilisation process or by unsatisfactory preparation of samples for cutting and mounting during the shock-freezing phase of sample preparation.

  9. Elemental distribution and sample integrity comparison of freeze-dried and frozen-hydrated biological tissue samples with nuclear microprobe

    Science.gov (United States)

    Vavpetič, P.; Vogel-Mikuš, K.; Jeromel, L.; Ogrinc Potočnik, N.; Pongrac, P.; Drobne, D.; Pipan Tkalec, Ž.; Novak, S.; Kos, M.; Koren, Š.; Regvar, M.; Pelicon, P.

    2015-04-01

    The analysis of biological samples in frozen-hydrated state with micro-PIXE technique at Jožef Stefan Institute (JSI) nuclear microprobe has matured to a point that enables us to measure and examine frozen tissue samples routinely as a standard research method. Cryotome-cut slice of frozen-hydrated biological sample is mounted between two thin foils and positioned on the sample holder. The temperature of the cold stage in the measuring chamber is kept below 130 K throughout the insertion of the samples and the proton beam exposure. Matrix composition of frozen-hydrated tissue is consisted mostly of ice. Sample deterioration during proton beam exposure is monitored during the experiment, as both Elastic Backscattering Spectrometry (EBS) and Scanning Transmission Ion Microscopy (STIM) in on-off axis geometry are recorded together with the events in two PIXE detectors and backscattered ions from the chopper in a single list-mode file. The aim of this experiment was to determine differences and similarities between two kinds of biological sample preparation techniques for micro-PIXE analysis, namely freeze-drying and frozen-hydrated sample preparation in order to evaluate the improvements in the elemental localisation of the latter technique if any. In the presented work, a standard micro-PIXE configuration for tissue mapping at JSI was used with five detection systems operating in parallel, with proton beam cross section of 1.0 × 1.0 μm2 and a beam current of 100 pA. The comparison of the resulting elemental distributions measured at the biological tissue prepared in the frozen-hydrated and in the freeze-dried state revealed differences in elemental distribution of particular elements at the cellular level due to the morphology alteration in particular tissue compartments induced either by water removal in the lyophilisation process or by unsatisfactory preparation of samples for cutting and mounting during the shock-freezing phase of sample preparation.

  10. Monitoring the marine environment using marine mammal tissue samples

    Energy Technology Data Exchange (ETDEWEB)

    Jones, P.D.; Hannah, D.J.; Day, P.J. [ESR:Environmental, Lower Hutt (New Zealand)] [and others

    1995-12-31

    Marine environments, both inshore and open ocean, receive numerous inputs of anthropogenic chemicals. Cetaceans provide a valuable resource for monitoring the low level contamination of marine environments with persistent organic contaminants. Comparative studies using inshore and offshore southern ocean cetaceans have revealed significant differences in the types of contamination in these two environments. The polychlorinated biphenyls (PCBs) deposited in the southern oceans are characterized by an abundance of lower chlorinated congeners. Polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F) are not present at significant concentrations in cetaceans from the open southern ocean. In contrast significant concentrations of PCDD/F congeners are detected in the blubber of the inshore living Hector`s dolphin. This species lives close to the shore and has a very small home range (approximately 30 km) for a cetacean. Analysis of tissue PCDD/F and PCB profiles from different populations and their food sources will be presented. The data are being used to determine if there are local variations in the contamination of the New Zealand inshore marine environment.

  11. Nested-PCR for the detection of Mycoplasma hyopneumoniae in bronchial alveolar swabs, frozen tissues and formalin-fixed paraffin-embedded swine lung samples: comparative evaluation with immunohistochemical findings and histological features

    Directory of Open Access Journals (Sweden)

    Paula R. Almeida

    2012-08-01

    Full Text Available The diagnosis of Mycoplasma hyopneumoniae infection is often performed through histopathology, immunohistochemistry (IHC and polymerase chain reaction (PCR or a combination of these techniques. PCR can be performed on samples using several conservation methods, including swabs, frozen tissue or formalin-fixed and paraffin-embedded (FFPE tissue. However, the formalin fixation process often inhibits DNA amplification. To evaluate whether M. hyopneumoniae DNA could be recovered from FFPE tissues, 15 lungs with cranioventral consolidation lesions were collected in a slaughterhouse from swine bred in herds with respiratory disease. Bronchial swabs and fresh lung tissue were collected, and a fragment of the corresponding lung section was placed in neutral buffered formalin for 48 hours. A PCR assay was performed to compare FFPE tissue samples with samples that were only refrigerated (bronchial swabs or frozen (tissue pieces. M. hyopneumoniae was detected by PCR in all 15 samples of the swab and frozen tissue, while it was detected in only 11 of the 15 FFPE samples. Histological features of M. hyopneumoniae infection were presented in 11 cases and 7 of these samples stained positive in IHC. Concordance between the histological features and detection results was observed in 13 of the FFPE tissue samples. PCR was the most sensitive technique. Comparison of different sample conservation methods indicated that it is possible to detect M. hyopneumoniae from FFPE tissue. It is important to conduct further research using archived material because the efficiency of PCR could be compromised under these conditions.

  12. First report of Streptococcus agalactiae isolated from Oreochromis niloticus in Piura, Peru: Molecular identification and histopathological lesions

    Directory of Open Access Journals (Sweden)

    Yessica Ortega Asencios

    2016-11-01

    Full Text Available The aim of this study was to identify the bacterium Streptococcus agalactiae isolated in farmed Nile tilapia (Oreochromis niloticus from Piura, Peru and to characterize the histopathological lesions caused by this pathogen. Sixteen tilapias were sampled with clinic signs of the disease such as erratic swimming, exophthalmia and haemorrhages on the body and fins. Qualitative PCR in real time and histopathological analysis were performed. Nine fishes positives to S. agalactiae were found. The main histopathological findings were fibrinosuppurative epicarditis, periesplenitis, meninigitis and panophtaltmitis with predominance of mononuclear infiltration in all tissues. The correlation between qPCR and histopathological findings demonstrated nine fish (prevalence of 56.25% with Cq lower than 30, associated to high degree of tissue injuries. This study reports the first isolation of S. agalactiae by PCR in real time in tilapia farmed in Peru and characterizes the major histopathological changes caused by this bacterium.

  13. Histopathological examination of Acanthamoeba sclerokeratitis

    Directory of Open Access Journals (Sweden)

    Kato K

    2014-01-01

    Full Text Available Kumiko Kato,1 Koji Hirano,2 Tetsuro Nagasaka,3 Koichi Matsunaga,1 Yuko Takashima,1 Mineo Kondo1 1Department of Ophthalmology, Mie University School of Medicine, Tsu, Japan; 2Department of Ophthalmology, Fujita Health University, Nagoya, Japan; 3Department of Medical Laboratory Science, Nagoya University Graduate School of Medicine, Nagoya, Japan Purpose: To report the histopathological findings in a case of severe Acanthamoeba sclerokeratitis (ASK. Patient and methods: A 46-year-old patient was referred to the Department of Ophthalmology of Mie University Hospital because of a severe corneal ulcer of the right eye of 6 months' duration. Our initial examination showed a ring-shaped corneal opacity with extensive epithelial defects and nodular scleritis. Cysts of Acanthamoeba were identified in cultures from corneal scrapings, and he was diagnosed with ASK. He was started on antiamoebic treatment, including topical micafungin and chlorhexidine. The corneal ulcer was debrided several times. One month later, he developed necrotizing scleritis, and the cornea suddenly perforated. The eye was enucleated because of severe pain and prepared for histopathological examination. Results: The histopathological examination showed an infiltration of polymorphonuclear leukocytes throughout the corneal stroma and also in the limbal area of the sclera, forming an abscess. Granulation tissue was observed in the anterior sclera close to the ciliary body, but the posterior regions of the eye were not affected by inflammation or tissue destruction. The cysts of Acanthamoeba were observed only in the cornea. Conclusion: Histopathological examination of an eye with severe ASK showed that the inflammation and tissue granulation were present only in the anterior part of eye, and the posterior segment was not affected. Because the inflammation and tissue destruction were confined to the anterior segment, enucleation might not have been necessary if the severe pain was

  14. Comparison of histopathology and real-time polymerase chain reaction (RT-PCR) for detection of Mycobacterium tuberculosis in fistula-in-ano.

    Science.gov (United States)

    Garg, Pankaj

    2017-07-01

    Histopathology is commonly used to diagnose tuberculosis in fistula-in-ano. The aim was to compare the sensitivity of polymerase chain reaction and histopathology in detecting tuberculosis in fistula-in-ano. The histopathology and polymerase chain-reaction of tissue (fistula tract) was done in all the consecutive operated cases. When pus sample was also available, polymerase chain reaction-pus was also done RESULTS: Three hundred forty seven samples (179 patients) were tested over 2 years (median 6.5 months). The mean age was 38.8 ± 10.7 years, and male/female was 170/9. Histopathology and polymerase chain reaction of tissue (fistula tract) was done in 152 and 165 patients, respectively. Polymerase chain reaction (pus) could be done in 30 patients. Overall, tuberculosis was detected in 20/179 (11.2%) patients. Of these, tuberculosis was detected by histopathology (tissue) in 1/152 (0.7%) and by polymerase chain reaction (tissue) in 14/165 (8.5%) patients. In pus, polymerase chain reaction detected tuberculosis in 6/30 (20%) patients. Both polymerase chain reaction of tissue and pus were positive in one patient. Polymerase chain reaction (tissue) and polymerase chain reaction (pus) were significantly more sensitive than histopathology (tissue) for detecting tuberculosis [histopathology 1/152 vs. polymerase chain reaction (tissue) 14/165, p = 0.0009] [histopathology 1/152 vs. polymerase chain reaction (pus) 6/30, p Polymerase chain reaction was significantly more sensitive than histopathology in detecting tuberculosis in fistula-in-ano. Histopathology might be missing out tuberculosis in many patients leading to recurrence of the fistula.

  15. Tissue Microarray Technology for Molecular Applications: Investigation of Cross-Contamination between Tissue Samples Obtained from the Same Punching Device.

    Science.gov (United States)

    Vassella, Erik; Galván, José A; Zlobec, Inti

    2015-04-02

    Tissue microarray (TMA) technology allows rapid visualization of molecular markers by immunohistochemistry and in situ hybridization. In addition, TMA instrumentation has the potential to assist in other applications: punches taken from donor blocks can be placed directly into tubes and used for nucleic acid analysis by PCR approaches. However, the question of possible cross-contamination between samples punched with the same device has frequently been raised but never addressed. Two experiments were performed. (1) A block from mycobacterium tuberculosis (TB) positive tissue and a second from an uninfected patient were aligned side-by-side in an automated tissue microarrayer. Four 0.6 mm punches were cored from each sample and placed inside their corresponding tube. Between coring of each donor block, a mechanical cleaning step was performed by insertion of the puncher into a paraffin block. This sequence of coring and cleaning was repeated three times, alternating between positive and negative blocks. A fragment from the 6110 insertion sequence specific for mycobacterium tuberculosis was analyzed; (2) Four 0.6 mm punches were cored from three KRAS mutated colorectal cancer blocks, alternating with three different wild-type tissues using the same TMA instrument (sequence of coring: G12D, WT, G12V, WT, G13D and WT). Mechanical cleaning of the device between each donor block was made. Mutation analysis by pyrosequencing was carried out. This sequence of coring was repeated manually without any cleaning step between blocks. In both analyses, all alternating samples showed the expected result (samples 1, 3 and 5: positive or mutated, samples 2, 4 and 6: negative or wild-type). Similar results were obtained without cleaning step. These findings suggest that no cross-contamination of tissue samples occurs when donor blocks are punched using the same device, however a cleaning step is nonetheless recommended. Our result supports the use of TMA technology as an accessory

  16. Tissue Microarray Technology for Molecular Applications: Investigation of Cross-Contamination between Tissue Samples Obtained from the Same Punching Device

    Directory of Open Access Journals (Sweden)

    Erik Vassella

    2015-04-01

    Full Text Available Background: Tissue microarray (TMA technology allows rapid visualization of molecular markers by immunohistochemistry and in situ hybridization. In addition, TMA instrumentation has the potential to assist in other applications: punches taken from donor blocks can be placed directly into tubes and used for nucleic acid analysis by PCR approaches. However, the question of possible cross-contamination between samples punched with the same device has frequently been raised but never addressed. Methods: Two experiments were performed. (1 A block from mycobacterium tuberculosis (TB positive tissue and a second from an uninfected patient were aligned side-by-side in an automated tissue microarrayer. Four 0.6 mm punches were cored from each sample and placed inside their corresponding tube. Between coring of each donor block, a mechanical cleaning step was performed by insertion of the puncher into a paraffin block. This sequence of coring and cleaning was repeated three times, alternating between positive and negative blocks. A fragment from the 6110 insertion sequence specific for mycobacterium tuberculosis was analyzed; (2 Four 0.6 mm punches were cored from three KRAS mutated colorectal cancer blocks, alternating with three different wild-type tissues using the same TMA instrument (sequence of coring: G12D, WT, G12V, WT, G13D and WT. Mechanical cleaning of the device between each donor block was made. Mutation analysis by pyrosequencing was carried out. This sequence of coring was repeated manually without any cleaning step between blocks. Results/Discussion: In both analyses, all alternating samples showed the expected result (samples 1, 3 and 5: positive or mutated, samples 2, 4 and 6: negative or wild-type. Similar results were obtained without cleaning step. These findings suggest that no cross-contamination of tissue samples occurs when donor blocks are punched using the same device, however a cleaning step is nonetheless recommended. Our

  17. Detection of Slit2 promoter hypermethylation in tissue and serum samples from breast cancer patients.

    Science.gov (United States)

    Kim, Ga-Eon; Lee, Kyung Hwa; Choi, Yoo Duk; Lee, Ji Shin; Lee, Jae Hyuk; Nam, Jong Hee; Choi, Chan; Park, Min Ho; Yoon, Jung Han

    2011-10-01

    Promoter hypermethylation has been shown to be a common mechanism for inactivation of tumor suppressor genes in breast cancer. The aim of this study was to investigate the prevalence of Slit2 promoter hypermethylation in both the tumor and serum samples of breast cancer patients with ductal carcinoma in situ (DCIS) or invasive breast carcinoma (IBC). The methylation status of Slit2 was investigated in 210 tissue samples (15 breast with no pathological findings, 26 DCIS, and 169 IBC samples) and 123 corresponding serum samples (15 breast with no pathological findings, 26 DCIS, and 82 IBC samples) using methylation-specific polymerase chain reaction. Immunohistochemical staining for Slit2 was also performed using tissue microarray blocks to determine whether Slit2 promoter hypermethylation correlated with loss of Slit2 expression. Slit2 promoter hypermethylation was not detected in breast tissue and serum samples from patients with no pathological findings. DCIS or IBC showed a statistically higher frequency of Slit2 promoter hypermethylation compared to breast with no pathological findings in both the tissue and serum samples; however, there were no statistically significant differences between DCIS and IBC samples. Similar Slit2 promoter hypermethylation patterns were seen in the tissue samples and corresponding serum specimens (p Slit2 promoter hypermethylation was associated with loss of Slit2 expression. These results suggest that Slit2 promoter hypermethylation appears to be responsible for functionally silencing Slit2 expression. Slit2 promoter hypermethylation may be considered as a possible serum marker for early detection of breast cancer.

  18. Clinical, Radiological, Microbiological, and Histopathological Aspects of Acquired Dacryocystoceles

    Directory of Open Access Journals (Sweden)

    Selam Yekta Sendul

    2014-01-01

    Full Text Available Purpose. The aim of this study is to investigate the etiology and the clinical, microbiological, histopathological, and radiological findings of acquired dacryocystoceles. Methods. In this retrospective study, we reviewed the clinical records of 10 eyes of 8 patients with dacryocystoceles who underwent external dacryocystorhinostomy (DCR surgery. Etiology, presenting symptoms and radiological findings as well as microbiological and histopathological assessment results and outcome were analyzed. Results. The records of 8 patients with dacryocystoceles were included in this study. In the histopathological evaluations of the samples collected from the lacrimal sac wall, chronic inflammation was found in all biopsied samples and fibrosis was observed in two histopathological evaluations. Computerized tomography (CT imaging showed fluid collection separated from adjacent tissues by a thin rim, corresponding to dacryocystoceles in the sac. In the microbiological culture examination of samples collected from the fluid within the cyst, no bacterial growth in 5 eyes, gram-negative bacillus growth in 3 eyes, and gram-positive cocci growth in 2 eyes were found. Conclusions. Acquired dacryocystoceles were observed extremely rarely and a definite pathogenic agent could not be identified in any of the cases, either microbiologically or histologically, whereas chronic inflammation was detected in all cases in our study.

  19. [American Badger Study : Sample Tissue Tracking Logs : Rocky Mountain Arsenal NWR : 1993

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This record contains the sample tissue tracking log data sheets related to the American Badger Study conducted at the Rocky Mountain Arsenal National Wildlife Refuge.

  20. Concentration of organochlorines in human brain, liver, and adipose tissue autopsy samples from Greenland

    DEFF Research Database (Denmark)

    Dewailly, Éric; Mulvad, Gert; Pedersen, Henning S.

    1999-01-01

    report results of organochlorine determination in liver, brain, omental fat, and subcutaneous abdominal fat samples collected from deceased Greenlanders between 1992 and 1994. Eleven chlorinated pesticides and 14 polychlorinated biphenyl congeners were measured in tissue lipid extracts by high...

  1. Effects of Re-heating Tissue Samples to Core Body Temperature on High-Velocity Ballistic Projectile-tissue Interactions.

    Science.gov (United States)

    Humphrey, Caitlin; Henneberg, Maciej; Wachsberger, Christian; Maiden, Nicholas; Kumaratilake, Jaliya

    2017-02-23

    Damage produced by high-speed projectiles on organic tissue will depend on the physical properties of the tissues. Conditioning organic tissue samples to human core body temperature (37°C) prior to conducting ballistic experiments enables their behavior to closely mimic that of living tissues. To minimize autolytic changes after death, the tissues are refrigerated soon after their removal from the body and re-heated to 37°C prior to testing. This research investigates whether heating 50-mm-cube samples of porcine liver, kidney, and heart to 37°C for varying durations (maximum 7 h) can affect the penetration response of a high-speed, steel sphere projectile. Longer conditioning times for heart and liver resulted in a slight loss of velocity/energy of the projectile, but the reverse effect occurred for the kidney. Possible reasons for these trends include autolytic changes causing softening (heart and liver) and dehydration causing an increase in density (kidney).

  2. Micro-Raman spectroscopy of tissue samples for oral pathology follow-up monitoring

    Science.gov (United States)

    Delfino, I.; Camerlingo, C.; Zenone, F.; Perna, G.; Capozzi, V.; Cirillo, N.; Gaeta, G. M.; Lepore, M.

    2010-04-01

    An "in vitro" study of Raman spectra from oral human tissues is reported in order to the develop a diagnostic method suitable for "in vivo" oral pathology follow-up. The investigated pathology is Pemphigus Vulgaris (PV) for which new techniques for guiding and monitoring therapy would be particularly useful. Raman spectra were obtained in the wavenumber regions from 1000 to 1800 cm-1 and 2700 to 3200 cm-1 from tissues from patients at different stages of pathology (active PV, under therapy and in PV remission stage) as confirmed by histopathological and immunofluorescence analysis. Differences in the spectra depending on tissue illness stage arise in 1150-1250 cm-1 (amide III) and 1420-1450 cm-1 (CH3 deformation) regions and around 1650 cm-1 (amide I) and 2930 cm-1 (CH3 symmetric stretch). A wavelet deconvolution procedure was applied to the spectra for better discriminating among the three different stages of illness and a linear regression analysis was used to fully exploit the content of information of Raman spectra.

  3. Mitochondrial Respiration Chain Enzymatic Activities in the Human Brain: Methodological Implications for Tissue Sampling and Storage.

    Science.gov (United States)

    Ronsoni, Marcelo Fernando; Remor, Aline Pertile; Lopes, Mark William; Hohl, Alexandre; Troncoso, Iris H Z; Leal, Rodrigo Bainy; Boos, Gustavo Luchi; Kondageski, Charles; Nunes, Jean Costa; Linhares, Marcelo Neves; Lin, Kátia; Latini, Alexandra Susana; Walz, Roger

    2016-04-01

    Mitochondrial respiratory chain complexes enzymatic (MRCCE) activities were successfully evaluated in frozen brain samples. Epilepsy surgery offers an ethical opportunity to study human brain tissue surgically removed to treat drug resistant epilepsies. Epilepsy surgeries are done with hemodynamic and laboratory parameters to maintain physiology, but there are no studies analyzing the association among these parameters and MRCCE activities in the human brain tissue. We determined the intra-operative parameters independently associated with MRCCE activities in middle temporal neocortex (Cx), amygdala (AMY) and head of hippocampus (HIP) samples of patients (n = 23) who underwent temporal lobectomy using multiple linear regressions. MRCCE activities in Cx, AMY and HIP are differentially associated to trans-operative mean arterial blood pressure, O2 saturation, hemoglobin, and anesthesia duration to time of tissue sampling. The time-course between the last seizure occurrence and tissue sampling as well as the sample storage to biochemical assessments were also associated with enzyme activities. Linear regression models including these variables explain 13-17 % of MRCCE activities and show a moderate to strong effect (r = 0.37-0.82). Intraoperative hemodynamic and laboratory parameters as well as the time from last seizure to tissue sampling and storage time are associated with MRCCE activities in human samples from the Cx, AMYG and HIP. Careful control of these parameters is required to minimize confounding biases in studies using human brain samples collected from elective neurosurgery.

  4. Heavy Metal Content in Thoracic Tissue Samples from Patients with and without NSCLC

    Directory of Open Access Journals (Sweden)

    Jessica Q. Tran

    2014-01-01

    Full Text Available Objectives. Environmental factors expose an individual to heavy metals that may stimulate cancer growth preclinically including non-small cell lung cancer (NSCLC cells. Here, we examine the prevalence of four heavy metals present in postsurgical tissues from individuals with and without NSCLC. Materials and Methods. Thoracic tissue samples from two separate sample sets were analyzed for cadmium (Cd, arsenic (As, mercury (Hg, and lead (Pb content. Results. In the first sample set, there was no significant measurable amount of Pb and Hg found in either NSCLC tissue or nonmalignant lung tissue samples. Cd was the most prevalent heavy metal and As was present in moderate amounts. In the second sample set, Cd was measurable across all tissue types taken from 28 NSCLC patients and significantly higher Cd was measurable in noncancer benign lung (n=9. In the NSCLC samples, As was measurable in moderate amounts, while Hg and Pb amounts were negligible. Conclusion. Cd and As are present in lung tissues for patients with NSCLC. With existing preclinical evidence of their tumorigenecity, it is plausible that Cd and/or As may have an impact on NSCLC development. Additional studies examining the prevalence and association between smokers and nonsmokers are suggested.

  5. Tissues and hair residues and histopathology in wild rats (Rattus rattus L.) and Algerian mice (Mus spretus Lataste) from an abandoned mine area (Southeast Portugal)

    Energy Technology Data Exchange (ETDEWEB)

    Pereira, R. [Departamento de Biologia da Universidade de Aveiro, Campus Universitario de Santiago, 3810-193 Aveiro (Portugal) and Instituto Piaget, Campus Academico de Viseu, Estrada do Alto do Gaio, Lordosa, 3515-776 Viseu (Portugal)]. E-mail: ruthp@bio.ua.pt; Pereira, M.L. [Departamento de Biologia da Universidade de Aveiro, Campus Universitario de Santiago, 3810-193 Aveiro (Portugal); Ribeiro, R. [Instituto do Ambiente e Vida, Departamento de Zoologia da Universidade de Coimbra, Largo Marques de Pombal, 3004-517 Coimbra (Portugal); Goncalves, F. [Departamento de Biologia da Universidade de Aveiro, Campus Universitario de Santiago, 3810-193 Aveiro (Portugal)

    2006-02-15

    Data gathered in this study suggested the exposure of rats and Algerian mice, living in an abandoned mining area, to a mixture of heavy metals. Although similar histopathological features were recorded in the liver and spleen of both species, the Algerian mouse has proved to be the strongest bioaccumulator species. Hair was considered to be a good biological material to monitor environmental contamination of Cr in rats. Significant positive associations were found between the levels of this element in hair/kidney (r = 0.826, n = 9, p < 0.01) and hair/liver (r = 0.697, n = 9, p = 0.037). Although no association was found between the levels of As recorded in the hair and in the organs, the levels of this element recorded in the hair, of both species, were significantly higher in animals captured in the mining area, which met the data from the organs analysed. Nevertheless, more studies will be needed to reduce uncertainty about cause-effect relationships. - The bioaccumulation of As and Cd and signs of renal histopathological injury proved the value of Algerian mice as a bioindicator species in the risk assessment of contaminated sites.

  6. Efficient and scalable serial extraction of DNA and RNA from frozen tissue samples.

    Science.gov (United States)

    Mathot, Lucy; Lindman, Monica; Sjöblom, Tobias

    2011-01-07

    Advances in cancer genomics have created a demand for scalable sample processing. We here present a process for serial extraction of nucleic acids from the same frozen tissue sample based on magnetic silica particles. The process is automation friendly with high recoveries of pure DNA and RNA suitable for analysis.

  7. Mitochondrial DNA from archived tissue samples kept in formalin for forensic odontology studies.

    Science.gov (United States)

    Pandey, Rahul; Mehrotra, Divya; Kowtal, Pradnya; Mahdi, Abbas Ali; Sarin, Rajiv

    2014-01-01

    Samples used for DNA isolation to be used for forensic odontology studies are often limited. The possibility to use tissue samples stored in formalin for a prolonged period, which contains nucleic acids of questionable quality, opens exciting possibilities for genetic and molecular biology studies useful in speciality of forensic odontology. The present study defines substantial modification of existing protocols for total genomic isolation including mitochondrial DNA and proves the utility of such obtained mitochondrial DNA in microsatellite analyses. 50 dental tissue samples which were kept in neutral buffered formalin liquid bottles were taken for DNA isolation and subsequent analysis. For the isolation of total genomic DNA from tissue samples, a new protocol with substantial modifications from routine ones was adopted by us. Total genomic DNA from matched blood samples were extracted using standard phenol-chloroform extraction method. Polymerase Chain Reaction and Sequencing of such extracted DNA samples for mitochondrial D loop region were successful and the results were comparable with DNA extracted from normal sources of samples. The present study reports for the first time that nucleic acids extracted from human dental tissue samples under prolonged formalin fixation times can be used for forensic odontology studies using the described methodology.

  8. The influence of the diagnostic technique on the histopathological diagnosis in malignant mesothelioma

    NARCIS (Netherlands)

    T. van Gelder (Teun); H.C. Hoogsteden (Henk); J.P. Vandenbroucke; Th.H. van der Kwast (Theo); H.T. Planteydt (Hans)

    1991-01-01

    textabstractIn the histopathology of malignant mesotheliomas three different types (epithelial, connective tissue and mixed type) are distinguished. Some authors believe all tumours to be of mixed type, but consider that due to inadequate sampling or small biopsies this may be missed frequently. In

  9. The influence of the diagnostic technique on the histopathological diagnosis in malignant mesothelioma

    NARCIS (Netherlands)

    T. van Gelder (Teun); H.C. Hoogsteden (Henk); J.P. Vandenbroucke; Th.H. van der Kwast (Theo); H.T. Planteydt (Hans)

    1991-01-01

    textabstractIn the histopathology of malignant mesotheliomas three different types (epithelial, connective tissue and mixed type) are distinguished. Some authors believe all tumours to be of mixed type, but consider that due to inadequate sampling or small biopsies this may be missed frequently. In

  10. Effect of sample volume size and sampling method on feline longitudinal myocardial velocity profiles from color tissue Doppler imaging.

    Science.gov (United States)

    Granström, Sara; Pipper, Christian Bressen; Møgelvang, Rasmus; Sogaard, Peter; Willesen, Jakob Lundgren; Koch, Jørgen

    2012-12-01

    The aims of this study were to compare the effect of sample volume (SV) size settings and sampling method on measurement variability and peak systolic (s'), and early (e') and late (a') diastolic longitudinal myocardial velocities using color tissue Doppler imaging (cTDI) in cats. Twenty cats with normal echocardiograms and 20 cats with hypertrophic cardiomyopathy. We quantified and compared empirical variance and average absolute values of s', e' and a' for three cardiac cycles using eight different SV settings (length 1,2,3 and 5 mm; width 1 and 2 mm) and three methods of sampling (end-diastolic sampling with manual tracking of the SV, end-systolic sampling without tracking, and random-frame sampling without tracking). No significant difference in empirical variance could be demonstrated between most of the tested SVs. However, the two settings with a length of 1 mm resulted in a significantly higher variance compared with all settings where the SV length exceeded 2 mm (p sampling method on the variability of measurements (p = 0.003) and manual tracking obtained the lowest variance. No difference in average values of s', e' or a' could be found between any of the SV settings or sampling methods. Within the tested range of SV settings, an SV length of 1 mm resulted in higher measurement variability compared with an SV length of 3 and 5 mm, and should therefore be avoided. Manual tracking of the sample volume is recommended. Copyright © 2012 Elsevier B.V. All rights reserved.

  11. Coupled analysis of in vitro and histology tissue samples to quantify structure-function relationship.

    Directory of Open Access Journals (Sweden)

    Evrim Acar

    Full Text Available The structure/function relationship is fundamental to our understanding of biological systems at all levels, and drives most, if not all, techniques for detecting, diagnosing, and treating disease. However, at the tissue level of biological complexity we encounter a gap in the structure/function relationship: having accumulated an extraordinary amount of detailed information about biological tissues at the cellular and subcellular level, we cannot assemble it in a way that explains the correspondingly complex biological functions these structures perform. To help close this information gap we define here several quantitative temperospatial features that link tissue structure to its corresponding biological function. Both histological images of human tissue samples and fluorescence images of three-dimensional cultures of human cells are used to compare the accuracy of in vitro culture models with their corresponding human tissues. To the best of our knowledge, there is no prior work on a quantitative comparison of histology and in vitro samples. Features are calculated from graph theoretical representations of tissue structures and the data are analyzed in the form of matrices and higher-order tensors using matrix and tensor factorization methods, with a goal of differentiating between cancerous and healthy states of brain, breast, and bone tissues. We also show that our techniques can differentiate between the structural organization of native tissues and their corresponding in vitro engineered cell culture models.

  12. Terahertz spectroscopy and detection of brain tumor in rat fresh-tissue samples

    Science.gov (United States)

    Yamaguchi, S.; Fukushi, Y.; Kubota, O.; Itsuji, T.; Yamamoto, S.; Ouchi, T.

    2015-03-01

    Terahertz (THz) spectroscopy and imaging of biomedical samples is expected to be an important application of THz analysis techniques. Identification and localization of tumor tissue, imaging of biological samples, and analysis of DNA by THz spectroscopy have been reported. THz time-domain spectroscopy (TDS) is useful for obtaining the refractive index over a broad frequency range. However, THz-TDS spectra of fresh tissue samples are sensitive to procedures such as sample preparation, and a standardized measurement protocol is required. Therefore, in this work, we establish a protocol for measurements of THz spectra of fresh tissue and demonstrate reliable detection of rat brain tumor tissue. We use a reflection THz-TDS system to measure the refractive index spectra of the samples mounted on a quartz plate. The tissue samples were measured immediately after sectioning to avoid sample denaturalization during storage. Special care was taken in THz data processing to eliminate parasitic reflections and reduce noise. The error level in our refractive index measurements was as low as 0.02 in the frequency range 0.8-1.5 THz. With increasing frequency, the refractive index in the tumor and normal regions monotonically decreased, similarly to water, and it was 0.02 higher in the tumor regions. The spectral data suggest that the tumor regions have higher water content. Hematoxylin-eosin stained images showed that increased cell density was also responsible for the observed spectral features. A set of samples from 10 rats showed consistent results. Our results suggest that reliable tumor detection in fresh tissue without pretreatment is possible with THz spectroscopy measurements. THz spectroscopy has the potential to become a real-time in vivo diagnostic method.

  13. MALDI direct analysis and imaging of frozen versus FFPE tissues: what strategy for which sample?

    Science.gov (United States)

    Wisztorski, Maxence; Franck, Julien; Salzet, Michel; Fournier, Isabelle

    2010-01-01

    Significant advances have been made in the past decade in the field of mass spectrometry imaging with MALDI ion sources (MALDI-MSI). While MALDI-MSI has high potential in the field of biology and in the clinic, a challenge for MALDI-MSI has been to adapt itself to a greater range of sample types. In particular, much of the biological archived materials for pathology studies are tissue biopsies fixed with paraformaldehyde and embedded in paraffin (FFPE tissues) because of the high stability of such samples. Thus, there has been a need to develop strategies for analyzing FFPE samples as this would allow retrospective studies of past clinical cases on large cohorts of existing samples. Obviously, PAF fixation, by inducing protein cross-linking, causes problems for molecular analysis by MS. We developed on tissue digestion strategies for overcoming these difficulties and allowing molecular data to be retrieved from FFPE samples no matter how long they have been stored. These digestion strategies preserve localization from digested proteins making MALDI-MSI of proteins possible by monitoring the resulting peptides. We present methods and protocols for FFPE samples. These strategies have proven to be valuable for all tested FFPE samples and have opened archived tissues from hospital banks to MALDI-MSI.

  14. Learning Histopathological Patterns

    DEFF Research Database (Denmark)

    Kårsnäs, Andreas; Dahl, Anders Lindbjerg; Larsen, Rasmus

    2011-01-01

    We propose a technique for analyzing images of immunohistochemically stained tissue samples for extracting features that correlate with patient disease. We address the problem of quantifying tumor tissue and segmenting and counting nuclei. Our method utilizes a flexible segmentation technique...... trained from representative image samples. Nuclei counting is based on a nucleus model that takes size, shape and nucleus probability into account. We obtain the probability of a nucleus from our segmentation procedure. Our method is experimentally validated on images stained with nuclear markers...

  15. Mucosal Incision and Forceps Biopsy for Reliable Tissue Sampling of Gastric Subepithelial Tumors

    Science.gov (United States)

    Shin, Sa Young; Lee, Sang Jin; Jun, Jae Hyuck; Park, Jong Kyu; Seo, Hyun Il; Han, Koon Hee; Kim, Young Don; Jeong, Woo Jin; Cheon, Gab Jin

    2017-01-01

    Background/Aims The diagnostic efficacy of current tissue sampling techniques for gastric subepithelial tumors (SETs) is limited. Better tissue sampling techniques are needed to improve pathological diagnosis. The aim of this study was to evaluate the safety and efficacy of a new technique, mucosal incision and forceps biopsy, for reliable tissue sampling of gastric SETs. Methods This study enrolled 12 consecutive patients who underwent mucosal incision and forceps biopsy of gastric SETs between November 2011 and September 2014 at Gangneung Asan Hospital. The medical records of patients were reviewed retrospectively. The safety and diagnostic yield of this method were evaluated. Results By performing mucosal incision and forceps biopsy, we were able to provide a definitive histological diagnosis for 11 out of 12 cases. The pathological diagnoses were leiomyoma (3/11), gastrointestinal stromal tumor (GIST; 2/11), lipoma (2/11), schwannoma (1/11), and ectopic pancreas (3/11). In cases of leiomyoma (n=3) and GIST (n=2), tissue samples were of sufficient size to allow immunohistochemical staining. In addition, the mitotic index was evaluated in two cases of GIST. There were no procedure-related complications. Conclusions Mucosal incision and forceps biopsy can be used as one of several methods to obtain adequate tissue samples from gastric SETs. PMID:26942580

  16. Broth versus solid agar culture of swab samples of cadaveric allograft musculoskeletal tissue.

    Science.gov (United States)

    Varettas, Kerry

    2013-12-01

    As part of the donor assessment protocol, bioburden assessment must be performed on allograft musculoskeletal tissue samples collected at the time of tissue retrieval. Swab samples of musculoskeletal tissue allografts from cadaveric donors are received at the microbiology department of the South Eastern Area Laboratory Services (Australia) to determine the presence of bacteria and fungi. This study will review the isolation rate of organisms from solid agar and broth culture of swab samples of cadaveric allograft musculoskeletal tissue over a 6-year period, 2006-2011. Swabs were inoculated onto horse blood agar (anaerobic, 35 °C) and chocolate agar (CO2, 35 °C) and then placed into a cooked meat broth (aerobic, 35 °C). A total of 1,912 swabs from 389 donors were received during the study period. 557 (29.1 %) swabs were culture positive with the isolation of 713 organisms, 249 (34.9 %) from solid agar culture and an additional 464 (65.1 %) from broth culture only. This study has shown that the broth culture of cadaveric allograft musculoskeletal swab samples recovered a greater amount of organisms than solid agar culture. Isolates such as Clostridium species and Staphylococcus aureus would not have been isolated from solid agar culture alone. Broth culture is an essential part of the bioburden assessment protocol of swab samples of cadaveric allograft musculoskeletal tissue in this laboratory.

  17. Impact of sample acquisition and linear amplification on gene expression profiling of lung adenocarcinoma: laser capture micro-dissection cell-sampling versus bulk tissue-sampling

    Directory of Open Access Journals (Sweden)

    Wigle Dennis A

    2009-03-01

    Full Text Available Abstract Background The methods used for sample selection and processing can have a strong influence on the expression values obtained through microarray profiling. Laser capture microdissection (LCM provides higher specificity in the selection of target cells compared to traditional bulk tissue selection methods, but at an increased processing cost. The benefit gained from the higher tissue specificity realized through LCM sampling is evaluated in this study through a comparison of microarray expression profiles obtained from same-samples using bulk and LCM processing. Methods Expression data from ten lung adenocarcinoma samples and six adjacent normal samples were acquired using LCM and bulk sampling methods. Expression values were evaluated for correlation between sample processing methods, as well as for bias introduced by the additional linear amplification required for LCM sample profiling. Results The direct comparison of expression values obtained from the bulk and LCM sampled datasets reveals a large number of probesets with significantly varied expression. Many of these variations were shown to be related to bias arising from the process of linear amplification, which is required for LCM sample preparation. A comparison of differentially expressed genes (cancer vs. normal selected in the bulk and LCM datasets also showed substantial differences. There were more than twice as many down-regulated probesets identified in the LCM data than identified in the bulk data. Controlling for the previously identified amplification bias did not have a substantial impact on the differences identified in the differentially expressed probesets found in the bulk and LCM samples. Conclusion LCM-coupled microarray expression profiling was shown to uniquely identify a large number of differentially expressed probesets not otherwise found using bulk tissue sampling. The information gain realized from the LCM sampling was limited to differential analysis

  18. X-ray scattering for the characterization of lyophilized breast tissue samples

    Science.gov (United States)

    Elshemey, Wael M.; Mohamed, Fayrouz S.; Khater, Ibrahim M.

    2013-09-01

    This work investigates the possibility of characterizing breast cancer by measuring the X-ray scattering profiles of lyophilized excised breast tissue samples. Since X-ray scattering from water-rich tissue is dominated by scattering from water, the removal of water by lyophilization would enhance the characterization process. In the present study, X-ray scattering profiles of 22 normal, 22 malignant and 10 benign breast tissue samples are measured. The cut-offs of scatter diagrams, sensitivity, specificity and diagnostic accuracy of three characterization parameters (full width at half maximum (FWHM) for the peak at 1.1 nm-1, area under curve (AUC), and ratio of 1st to 2nd scattering peak intensities (I1/I2%)) are calculated and compared to the data from non-lyophilized samples. Results show increased sensitivity (up to 100%) of the present data on lyophilized breast tissue samples compared to previously reported data for non-lyophilized samples while the specificity (up to 95.4%), diagnostic accuracy (up to 95.4%) and receiver operating characteristic (ROC) curve values (up to 0.9979) for both sets of data are comparable. The present study shows significant differences between normal samples and each of malignant and benign samples. Only subtle differences exist between malignant and benign lyophilized breast tissue samples where FWHM=0.7±0.1 and 0.8±0.3, AUC=1.3±0.2 and 1.4±0.2 and I1/I2%=44.9±11.0 and 52.4±7.6 for malignant and benign samples respectively.

  19. Measurement of the hyperelastic properties of 44 pathological ex vivo breast tissue samples

    Science.gov (United States)

    O'Hagan, Joseph J.; Samani, Abbas

    2009-04-01

    The elastic and hyperelastic properties of biological soft tissues have been of interest to the medical community. There are several biomedical applications where parameters characterizing such properties are critical for a reliable clinical outcome. These applications include surgery planning, needle biopsy and brachtherapy where tissue biomechanical modeling is involved. Another important application is interpreting nonlinear elastography images. While there has been considerable research on the measurement of the linear elastic modulus of small tissue samples, little research has been conducted for measuring parameters that characterize the nonlinear elasticity of tissues included in tissue slice specimens. This work presents hyperelastic measurement results of 44 pathological ex vivo breast tissue samples. For each sample, five hyperelastic models have been used, including the Yeoh, N = 2 polynomial, N = 1 Ogden, Arruda-Boyce, and Veronda-Westmann models. Results show that the Yeoh, polynomial and Ogden models are the most accurate in terms of fitting experimental data. The results indicate that almost all of the parameters corresponding to the pathological tissues are between two times to over two orders of magnitude larger than those of normal tissues, with C11 showing the most significant difference. Furthermore, statistical analysis indicates that C02 of the Yeoh model, and C11 and C20 of the polynomial model have very good potential for cancer classification as they show statistically significant differences for various cancer types, especially for invasive lobular carcinoma. In addition to the potential for use in cancer classification, the presented data are very important for applications such as surgery planning and virtual reality based clinician training systems where accurate nonlinear tissue response modeling is required.

  20. Effects of formalin fixation on tissue optical properties of in-vitro brain samples

    Science.gov (United States)

    Anand, Suresh; Cicchi, Riccardo; Martelli, Fabrizio; Giordano, Flavio; Buccoliero, Anna Maria; Guerrini, Renzo; Pavone, Francesco S.

    2015-03-01

    Application of light spectroscopy based techniques for the detection of cancers have emerged as a promising approach for tumor diagnostics. In-vivo or freshly excised samples are normally used for point spectroscopic studies. However, ethical issues related to in-vivo studies, rapid decay of surgically excised tissues and sample availability puts a limitation on in-vivo and in-vitro studies. There has been a few studies reported on the application of formalin fixed samples with good discrimination capability. Usually formalin fixation is performed to prevent degradation of tissues after surgical resection. Fixing tissues in formalin prevents cell death by forming cross-linkages with proteins. Previous investigations have revealed that washing tissues fixed in formalin using phosphate buffered saline is known to reduce the effects of formalin during spectroscopic measurements. But this could not be the case with reflectance measurements. Hemoglobin is a principal absorbing medium in biological tissues in the visible range. Formalin fixation causes hemoglobin to seep out from red blood cells. Also, there could be alterations in the refractive index of tissues when fixed in formalin. In this study, we propose to investigate the changes in tissue optical properties between freshly excised and formalin fixed brain tissues. The results indicate a complete change in the spectral profile in the visible range where hemoglobin has its maximum absorption peaks. The characteristic bands of oxy-hemoglobin at 540, 580 nm and deoxy-hemoglobin at 555 nm disappear in the case of samples fixed in formalin. In addition, an increased spectral intensity was observed for the wavelengths greater than 650 nm where scattering phenomena are presumed to dominate.

  1. Quantitative mapping of collagen fiber alignment in thick tissue samples using transmission polarized-light microscopy.

    Science.gov (United States)

    Yakovlev, Dmitry D; Shvachkina, Marina E; Sherman, Maria M; Spivak, Andrey V; Pravdin, Alexander B; Yakovlev, Dmitry A

    2016-07-01

    Immersion optical clearing makes it possible to use transmission polarized-light microscopy for characterization of thick (200 to 2000  μm) layers of biological tissues. We discuss polarization properties of thick samples in the context of the problem of characterization of collagen fiber alignment in connective tissues such as sclera and dermis. Optical chirality caused by azimuthal variations of the macroscopic (effective) optic axis of the medium across the sample thickness should be considered in polarization mapping of thick samples of these tissues. We experimentally evaluate to what extent the optical chirality affects the measurement results in typical situations and show under what conditions it can be easily taken into account and does not hinder, but rather helps, in characterization of collagen fiber alignment.

  2. Sample Preparation of Corn Seed Tissue to Prevent Analyte Relocations for Mass Spectrometry Imaging

    Science.gov (United States)

    Kim, Shin Hye; Kim, Jeongkwon; Lee, Young Jin; Lee, Tae Geol; Yoon, Sohee

    2017-08-01

    Corn seed tissue sections were prepared by the tape support method using an adhesive tape, and mass spectrometry imaging (MSI) was performed. The effect of heat generated during sample preparation was investigated by time-of-flight secondary mass spectrometry (TOF-SIMS) imaging of corn seed tissue prepared by the tape support and the thaw-mounted methods. Unlike thaw-mounted sample preparation, the tape support method does not cause imaging distortion because of the absence of heat, which can cause migration of the analytes on the sample. By applying the tape-support method, the corn seed tissue was prepared without structural damage and MSI with accurate spatial information of analytes was successfully performed.

  3. Trace element contamination in feather and tissue samples from Anna’s hummingbirds

    Science.gov (United States)

    Mikoni, Nicole A.; Poppenga, Robert H.; Ackerman, Joshua T.; Foley, Janet E.; Hazlehurst, Jenny; Purdin, Güthrum; Aston, Linda; Hargrave, Sabine; Jelks, Karen; Tell, Lisa A.

    2017-01-01

    Trace element contamination (17 elements; Be, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Mo, Cd, Ba, Hg, Tl, and Pb) of live (feather samples only) and deceased (feather and tissue samples) Anna's hummingbirds (Calypte anna) was evaluated. Samples were analyzed using inductively coupled plasma-mass spectrometry (ICP-MS; 17 elements) and atomic absorption spectrophotometry (Hg only). Mean plus one standard deviation (SD) was considered the benchmark, and concentrations above the mean + 1 SD were considered elevated above normal. Contour feathers were sampled from live birds of varying age, sex, and California locations. In order to reduce thermal impacts, minimal feathers were taken from live birds, therefore a novel method was developed for preparation of low mass feather samples for ICP-MS analysis. The study found that the novel feather preparation method enabled small mass feather samples to be analyzed for trace elements using ICP-MS. For feather samples from live birds, all trace elements, with the exception of beryllium, had concentrations above the mean + 1 SD. Important risk factors for elevated trace element concentrations in feathers of live birds were age for iron, zinc, and arsenic, and location for iron, manganese, zinc, and selenium. For samples from deceased birds, ICP-MS results from body and tail feathers were correlated for Fe, Zn, and Pb, and feather concentrations were correlated with renal (Fe, Zn, Pb) or hepatic (Hg) tissue concentrations. Results for AA spectrophotometry analyzed samples from deceased birds further supported the ICP-MS findings where a strong correlation between mercury concentrations in feather and tissue (pectoral muscle) samples was found. These study results support that sampling feathers from live free-ranging hummingbirds might be a useful, non-lethal sampling method for evaluating trace element exposure and provides a sampling alternative since their small body size limits traditional sampling of blood and tissues. The

  4. Sample processing considerations for detecting copy number changes in formalin-fixed, paraffin-embedded tissues.

    Science.gov (United States)

    Jacobs, Sharoni

    2012-11-01

    The Whole Genome Sampling Analysis (WGSA) assay in combination with Affymetrix GeneChip Mapping Arrays is used for copy number analysis of high-quality DNA samples (i.e., samples that have been collected from blood, fresh or frozen tissue, or cell lines). Formalin-fixed, paraffin-embedded (FFPE) samples, however, represent the most prevalent form of archived clinical samples, but they provide additional challenges for molecular assays. FFPE processing usually results in the degradation of FFPE DNA and in the contamination and chemical modification of these DNA samples. Because of these issues, FFPE DNA is not suitable for all molecular assays designed for high-quality DNA samples. Strategies recommended for processing FFPE DNA samples through WGSA and to the Mapping arrays are described here.

  5. Cancer Detection in Human Tissue Samples Using a Fiber-Tip pH Probe.

    Science.gov (United States)

    Schartner, Erik P; Henderson, Matthew R; Purdey, Malcolm; Dhatrak, Deepak; Monro, Tanya M; Gill, P Grantley; Callen, David F

    2016-12-01

    Intraoperative detection of tumorous tissue is an important unresolved issue for cancer surgery. Difficulty in differentiating between tissue types commonly results in the requirement for additional surgeries to excise unremoved cancer tissue or alternatively in the removal of excess amounts of healthy tissue. Although pathologic methods exist to determine tissue type during surgery, these methods can compromise postoperative pathology, have a lag of minutes to hours before the surgeon receives the results of the tissue analysis, and are restricted to excised tissue. In this work, we report the development of an optical fiber probe that could potentially find use as an aid for margin detection during surgery. A fluorophore-doped polymer coating is deposited on the tip of an optical fiber, which can then be used to record the pH by monitoring the emission spectra from this dye. By measuring the tissue pH and comparing with the values from regular tissue, the tissue type can be determined quickly and accurately. The use of a novel lift-and-measure technique allows for these measurements to be performed without influence from the inherent autofluorescence that commonly affects fluorescence-based measurements on biological samples. The probe developed here shows strong potential for use during surgery, as the probe design can be readily adapted to a low-cost portable configuration, which could find use in the operating theater. Use of this probe in surgery either on excised or in vivo tissue has the potential to improve success rates for complete removal of cancers. Cancer Res; 76(23); 6795-801. ©2016 AACR.

  6. [Study of regeneration in periodontal tissue after implantation of bone ceramic and collagen gel compound materials. Evaluation of histopathological finding and autoradiography].

    Science.gov (United States)

    Miyamoto, Y; Hayashi, H; Kamoi, K

    1989-12-01

    The aim of this study is to determine the process of periodontal tissue regeneration and the metabolic activity of osteoblasts after implantation of bone ceramic and collagen gel compound materials (BC). Bone defects were artificially prepared in the alveolar septa of the bilateral upper first and second molars of Wistar rats. Subsequently, BC were implanted into the defective sites on the left side, and the gingival flaps were closed. At the defective sites on the right side, as a control, gingival flaps were closed without implantation. Rats were sacrificed 1, 3, 5, 7 or 14 weeks after implantation, and prepared tissue sections were observed both pathologically and autoradiographically using 3H-Proline. The results obtained were as follows: Pathological Findings One week after BC implantation, inflammatory cellular infiltration of the surrounding gingival connective tissue was relatively mild. Three weeks after implantation, BC were present in fibrous connective tissues, and some directly bound to the marices of regenerated bone. Observation 5 weeks after implantation revealed that BC had become embedded in the regenerated bone matrices and that there was giant cell reaction to foreign bodies at the margin of BC located in connective tissue. BC were directly bound to the regenerated bone matrices without intermediary fibrous tissues 7 and 14 weeks after implantation. Connective tissues showed high grade regeneration of collagen fiber bundles, in an arrangement that tended to be fixed in mesial and distal directions. Autoradiographic Findings There was no uptake of 3H-Proline into the regenerated bone matrices or the gingival connective tissue surrounding BC, while uptake of 3H-Proline into the entire area around the root apex and in the vicinity of the alveolar septum was observed with time (weeks) after BC implantation. These results suggest that BC provide nuclei for bone regeneration through inclusion in newly-generated periodontal bone tissue, although it is

  7. Collecting and Storing Tissue, Blood, and Bone Marrow Samples From Patients With Rhabdomyosarcoma or Other Soft Tissue Sarcoma

    Science.gov (United States)

    2017-09-26

    Adult Rhabdomyosarcoma; Childhood Desmoplastic Small Round Cell Tumor; Chordoma; Desmoid Tumor; Metastatic Childhood Soft Tissue Sarcoma; Nonmetastatic Childhood Soft Tissue Sarcoma; Previously Treated Childhood Rhabdomyosarcoma; Previously Untreated Childhood Rhabdomyosarcoma; Recurrent Adult Soft Tissue Sarcoma; Recurrent Childhood Rhabdomyosarcoma; Recurrent Childhood Soft Tissue Sarcoma; Stage I Adult Soft Tissue Sarcoma; Stage II Adult Soft Tissue Sarcoma; Stage III Adult Soft Tissue Sarcoma; Stage IV Adult Soft Tissue Sarcoma

  8. Automated MALDI Matrix Coating System for Multiple Tissue Samples for Imaging Mass Spectrometry

    Science.gov (United States)

    Mounfield, William P.; Garrett, Timothy J.

    2012-03-01

    Uniform matrix deposition on tissue samples for matrix-assisted laser desorption/ionization (MALDI) is key for reproducible analyte ion signals. Current methods often result in nonhomogenous matrix deposition, and take time and effort to produce acceptable ion signals. Here we describe a fully-automated method for matrix deposition using an enclosed spray chamber and spray nozzle for matrix solution delivery. A commercial air-atomizing spray nozzle was modified and combined with solenoid controlled valves and a Programmable Logic Controller (PLC) to control and deliver the matrix solution. A spray chamber was employed to contain the nozzle, sample, and atomized matrix solution stream, and to prevent any interference from outside conditions as well as allow complete control of the sample environment. A gravity cup was filled with MALDI matrix solutions, including DHB in chloroform/methanol (50:50) at concentrations up to 60 mg/mL. Various samples (including rat brain tissue sections) were prepared using two deposition methods (spray chamber, inkjet). A linear ion trap equipped with an intermediate-pressure MALDI source was used for analyses. Optical microscopic examination showed a uniform coating of matrix crystals across the sample. Overall, the mass spectral images gathered from tissues coated using the spray chamber system were of better quality and more reproducible than from tissue specimens prepared by the inkjet deposition method.

  9. Citrulline as a Biomarker in the Murine Total-Body Irradiation Model: Correlation of Circulating and Tissue Citrulline to Small Intestine Epithelial Histopathology.

    Science.gov (United States)

    Jones, Jace W; Tudor, Gregory; Li, Fei; Tong, Yan; Katz, Barry; Farese, Ann M; MacVittie, Thomas J; Booth, Catherine; Kane, Maureen A

    2015-11-01

    The use of plasma citrulline as a biomarker for gastrointestinal acute radiation syndrome via exposure to total-body irradiation in a murine model was investigated. The radiation exposure covered lethal, mid-lethal, and sub-lethal gastrointestinal acute radiation syndrome. Plasma citrulline profiles were generated over the first 6 d following total-body irradiation exposure of 6-15 Gy. In addition, plasma citrulline was comprehensively evaluated in the context of matching small intestine citrulline and histopathology. Higher plasma citrulline was significantly associated with lower irradiation doses over the first 6 d following the irradiation insult. Furthermore, higher plasma citrulline was significantly associated with higher crypt survival. The correlation of the plasma citrulline to crypt survival was more robust for higher irradiation doses and for later time points. The data suggested plasma citrulline was most informative for reflecting gastrointestinal injury resulting from exposure to 9-15 Gy total-body irradiation covering time-points 2-5 d post the irradiation insult.

  10. [Detection of bovine leukaemia virus (BLV) in tissue samples of naturally and experimentally infected cattle].

    Science.gov (United States)

    Teifke, Jens P; Vahlenkamp, Thomas W

    2008-01-01

    Enzootic bovine leukaemia (EBL) which is caused by the bovine leukaemia virus (BLV) still plays a remarkable role despite a significant success in sanitation programmes. In the Federal Republic of Germany it was not possible to eradicate the disease until today. Sporadically during slaughter or necropsy of cattle neoplastic lesions of the lymphatic tissues are observed that need to be clarified with regard to BLV as etiological agent. Due to the fact that in most instances no serological data are available from the respective animals and blood drawings from the original holdings are not easy to obtain the polymerase chain reaction (PCR) opens new avenues as supplementary diagnostic tool to test unfixed lymphatic tissues for the presence of BLV proviral DNA. Lymph node tissues from 10 naturally or experimentally BLV-infected cattle, which have been monitored virologically and serologically, and tissues from 4 negative animals were processed, DNA was extracted and subjected to PCR to amplify BLV env gene specific sequences. The results show that in cattle with BLV-induced leukosis as well as in cattle, which were clinically healthy and unsuspicious at slaughter or at post-mortem, either with persistent lymphocytosis (PL) or without, BLV proviral DNA could be detected easily in samples of lymphatic tissues and in high concordance with serological data. In this article data from the National and OIE reference laboratory for EBL at the Friedrich-Loeffler-Institut (FLI, Germany) are presented. Elaborated laboratory protocols for processing of tissue samples and performing of BLV-PCR are recommended.

  11. Semiquantitative determination of polychlorinated biphenyls in tissue samples by thin layer chromatography

    Science.gov (United States)

    Mulhern, B.M.; Cromartie, E.; Reichel, W.L.; Belisle, A.A.

    1971-01-01

    A method is described for the analysis of polychlorinated biphenyl (PCB) compounds in tissue samples. Cleanup by hexane-aceto-nitrile partitioning and Florisil column chromatography are performed on samples before oxidative treatment to convert DDE to DCBP. PCB components are then determined semi-quantitatively by TLC. No prior separation of PCB from chlorinated pesticides is required. The lower limit of sensitivity is 0.2 ?g.

  12. Three dimensional imaging of paraffin embedded human lung tissue samples by micro-computed tomography.

    Directory of Open Access Journals (Sweden)

    Anna E Scott

    Full Text Available Understanding the three-dimensional (3-D micro-architecture of lung tissue can provide insights into the pathology of lung disease. Micro computed tomography (µCT has previously been used to elucidate lung 3D histology and morphometry in fixed samples that have been stained with contrast agents or air inflated and dried. However, non-destructive microstructural 3D imaging of formalin-fixed paraffin embedded (FFPE tissues would facilitate retrospective analysis of extensive tissue archives of lung FFPE lung samples with linked clinical data.FFPE human lung tissue samples (n = 4 were scanned using a Nikon metrology µCT scanner. Semi-automatic techniques were used to segment the 3D structure of airways and blood vessels. Airspace size (mean linear intercept, Lm was measured on µCT images and on matched histological sections from the same FFPE samples imaged by light microscopy to validate µCT imaging.The µCT imaging protocol provided contrast between tissue and paraffin in FFPE samples (15 mm x 7 mm. Resolution (voxel size 6.7 µm in the reconstructed images was sufficient for semi-automatic image segmentation of airways and blood vessels as well as quantitative airspace analysis. The scans were also used to scout for regions of interest, enabling time-efficient preparation of conventional histological sections. The Lm measurements from µCT images were not significantly different to those from matched histological sections.We demonstrated how non-destructive imaging of routinely prepared FFPE samples by laboratory µCT can be used to visualize and assess the 3D morphology of the lung including by morphometric analysis.

  13. Concentration of organochlorines in human brain, liver, and adipose tissue autopsy samples from Greenland

    DEFF Research Database (Denmark)

    Dewailly, Éric; Mulvad, Gert; Pedersen, Henning S.

    1999-01-01

    Organochlorines are persistent lipophilic compounds that accumulate in Inuit people living in circumpolar countries. Organochlorines accumulate as a result of the Inuits' large consumption of sea mammal fat; however, available data are limited to blood lipids, milk fat, and adipose tissue. We...... report results of organochlorine determination in liver, brain, omental fat, and subcutaneous abdominal fat samples collected from deceased Greenlanders between 1992 and 1994. Eleven chlorinated pesticides and 14 polychlorinated biphenyl congeners were measured in tissue lipid extracts by high......-resolution gas chromatography with electron capture detection. Mean concentrations of polychlorinated biphenyls, 2, 2'-bis(4-chlorophenyl)-1,1-dichloroethylene, ss-hexachlorocyclohexane, hexachlorobenzene, mirex, trans-nonachlor, and oxychlordane in adipose tissue samples from Greenlanders were 3-34-fold higher...

  14. DNA damage in preserved specimens and tissue samples: a molecular assessment

    Directory of Open Access Journals (Sweden)

    Cantin Elizabeth

    2008-10-01

    Full Text Available Abstract The extraction of genetic information from preserved tissue samples or museum specimens is a fundamental component of many fields of research, including the Barcode of Life initiative, forensic investigations, biological studies using scat sample analysis, and cancer research utilizing formaldehyde-fixed, paraffin-embedded tissue. Efforts to obtain genetic information from these sources are often hampered by an inability to amplify the desired DNA as a consequence of DNA damage. Previous studies have described techniques for improved DNA extraction from such samples or focused on the effect of damaging agents – such as light, oxygen or formaldehyde – on free nucleotides. We present ongoing work to characterize lesions in DNA samples extracted from preserved specimens. The extracted DNA is digested to single nucleosides with a combination of DNase I, Snake Venom Phosphodiesterase, and Antarctic Phosphatase and then analyzed by HPLC-ESI-TOF-MS. We present data for moth specimens that were preserved dried and pinned with no additional preservative and for frog tissue samples that were preserved in either ethanol, or formaldehyde, or fixed in formaldehyde and then preserved in ethanol. These preservation methods represent the most common methods of preserving animal specimens in museum collections. We observe changes in the nucleoside content of these samples over time, especially a loss of deoxyguanosine. We characterize the fragmentation state of the DNA and aim to identify abundant nucleoside lesions. Finally, simple models are introduced to describe the DNA fragmentation based on nicks and double-strand breaks.

  15. The Novel Application of Non-Lethal Citizen Science Tissue Sampling in Recreational Fisheries.

    Directory of Open Access Journals (Sweden)

    Samuel M Williams

    Full Text Available Increasing fishing pressure and uncertainty surrounding recreational fishing catch and effort data promoted the development of alternative methods for conducting fisheries research. A pilot investigation was undertaken to engage the Australian game fishing community and promote the non-lethal collection of tissue samples from the black marlin Istiompax indica, a valuable recreational-only species in Australian waters, for the purpose of future genetic research. Recruitment of recreational anglers was achieved by publicizing the project in magazines, local newspapers, social media, blogs, websites and direct communication workshops at game fishing tournaments. The Game Fishing Association of Australia and the Queensland Game Fishing Association were also engaged to advertise the project and recruit participants with a focus on those anglers already involved in the tag-and-release of marlin. Participants of the program took small tissue samples using non-lethal methods which were stored for future genetic analysis. The program resulted in 165 samples from 49 participants across the known distribution of I. indica within Australian waters which was a sufficient number to facilitate a downstream population genetic analysis. The project demonstrated the potential for the development of citizen science sampling programs to collect tissue samples using non-lethal methods in order to achieve targeted research objects in recreationally caught species.

  16. Histopathological finding of liver and kidney tissues of the yellow mystus, Hemibagrus filamentus (Fang and Chaux, 1949, from the Tapee River, Thailand

    Directory of Open Access Journals (Sweden)

    Sinlapachai Senarat

    2015-02-01

    Full Text Available A histological finding of liver and kidney tissues of yellow mystus, Hemibagrus filamentus (Fang and Chaux, 1949, collected from the Tapee River (Chawang District, Nakhon Si Thammarat Province, Thailand was conducted to examine the pathological lesions. Lesions, including cellular swelling, hydropic degeneration, or vacuolation of hepatocytes were found. From this, coagulative necrosis with pyknotic nuclei, infiltration of macrophage, constriction of sinusoidal capillaries and damage of endothelial cells of blood vessels were observed in liver tissues. The dilation of both Bowman’s space and the glomerulus were observed from kidney lesions. Moreover, necrosis in renal tubular cells was also observed in some areas. Although preliminary, the results from this study clearly show the need for further investigation into stress factors and vectors affecting the health of H. filamentus and other fish species within the Tapee River.

  17. Comparison of different biopsy forceps models for tissue sampling in eosinophilic esophagitis.

    Science.gov (United States)

    Bussmann, Christian; Schoepfer, Alain M; Safroneeva, Ekaterina; Haas, Nadine; Godat, Sébastien; Sempoux, Christine; Simon, Hans-Uwe; Straumann, Alex

    2016-12-01

    Background and aims: Eosinophilic esophagitis (EoE) is a mixed inflammatory and fibrostenotic disease. Unlike superficial inflammatory changes, subepithelial fibrosis is not routinely sampled in esophageal biopsies. This study aimed to evaluate the efficacy and safety of deep esophageal sampling with four different types of biopsy forceps. Patients and methods: In this cross-sectional study, esophageal biopsies were taken in 30 adult patients by one expert endoscopist. Biopsies sampled from distal esophagus using a static jaw forceps (Olympus, FB-11K-1) were compared with proximal biopsies sampled with static jaw (Olympus, FB-45Q-1), alligator jaw (Olympus, FB-210K), and large-capacity forceps (Boston Scientific, Radial Jaw 4). One pathologist calculated the surface area of epithelial and subepithelial layers in hematoxylin and eosin (H&E)-stained biopsies. Results: Subepithelial tissue was acquired in 97 % (static jaw FB-11K-1), 93 % (static jaw FB-45Q-1), 80 % (alligator jaw), and 55 % (large-capacity) of samples. Median (interquartile [IQR]) ratios of surface area of epithelial to subepithelial tissue were: static jaw FB-45Q-1, 1.07 (0.65 - 4.465); static jaw FB-11K-1, 1.184 (0.608 - 2.545); alligator jaw, 2.353 (1.312 - 4.465); and large-capacity, 2.71 (1.611 - 4.858). The static jaw models obtained a larger surface area of subepithelial tissue compared with the alligator jaw (P  90 % of biopsies and appear to be superior to alligator or large-capacity forceps in sampling larger amounts of subepithelial tissue. © Georg Thieme Verlag KG Stuttgart · New York.

  18. Imaging of Proteins in Tissue Samples Using Nanospray Desorption Electrospray Ionization Mass Spectrometry.

    Science.gov (United States)

    Hsu, Cheng-Chih; Chou, Pi-Tai; Zare, Richard N

    2015-11-17

    Chemical maps of tissue samples provide important information on biological processes therein. Recently, advances in tissue imaging have been achieved using ambient ionization techniques, such as desorption electrospray ionization mass spectrometry (DESI-MS), but such techniques have been almost exclusively confined to the mapping of lipids and metabolites. We report here the use of nanospray desorption electrospray ionization (nanoDESI) that allows us to image proteins in tissue samples in a label-free manner at atmospheric pressure with only minimum sample preparation. Multiply charged proteins with masses up to 15 kDa were successfully detected by nanoDESI using an LTQ Orbitrap mass spectrometer. In an adult mice brain section, expression of proteins including ubiquitin, β-thymosin, myelin basic protein, and hemoglobin were spatially mapped and characterized. We also determined the location of methylation on myelin basic protein. This imaging modality was further implemented to MYC-induced lymphomas. We observed an array of truncated proteins in the region where normal thymus cells were infiltrated by tumor cells, in contrast to healthy tissue.

  19. Do anesthetics and sampling strategies affect transcription analysis of fish tissues?

    Directory of Open Access Journals (Sweden)

    Hevrøy Ernst M

    2007-06-01

    Full Text Available Abstract Background The aim of the current examination was to evaluate if sedation and anesthetic treatment techniques affect the quality of RNA extracted from liver, gill, head kidney and brain tissues in Atlantic salmon Salmo salar L. Blood parameters were measured and tissue specimens sampled in six groups of fish; one control group (0 minutes, two groups kept in pure seawater in 90 liter tanks for 30 and 120 minutes, two groups treated with the anesthetic isoeugenol for 30 and 120 minutes, and one group kept in pure seawater for 105 minutes and then anaesthetized with metacaine for 15 minutes. RNA quality was assessed with the NanoDrop ND-1000 spectrophotometer (260/280 and 260/230 nm ratios and with the Agilent Bioanalyzer (28S/18S ratio and RIN data in samples either preserved in liquefied nitrogen (N2 or in RNAlater. In addition, the transcriptional levels of two fast-responding genes were quantified in gill and brain tissues. Results The results show that physiological stress during sampling does not affect the quality of RNA extracted from fish specimens. However, prolonged sedation (2 hours resulted in a metabolic alkalosis that again affected the transcriptional levels of genes involved in ionoregulation and respiration. In gills, Na+-K+-ATPase α1b was significantly downregulated and hypoxia inducible factor 1 (HIF1 significantly upregulated after two hours of treatment with isoeugenol, suggesting that this commonly used sedative affects osmo-regulation and respiration in the fish. The results also suggest that for tissue preservation in general it is better to flash-freeze fish specimens in liquefied N2 than to use RNAlater. Conclusion Prolonged sedation may affect the transcription of fast-responding genes in tissues of fish. Two hours of sedation with isoeugenol resulted in downregulation of the Na+-K+-ATPase α1b gene and upregulation of the HIF1 gene in gills of Atlantic salmon. The quality of RNA extracted from tissue specimens

  20. Spatially-Resolved Proteomics: Rapid Quantitative Analysis of Laser Capture Microdissected Alveolar Tissue Samples

    Energy Technology Data Exchange (ETDEWEB)

    Clair, Geremy; Piehowski, Paul D.; Nicola, Teodora; Kitzmiller, Joseph A.; Huang, Eric L.; Zink, Erika M.; Sontag, Ryan L.; Orton, Daniel J.; Moore, Ronald J.; Carson, James P.; Smith, Richard D.; Whitsett, Jeffrey A.; Corley, Richard A.; Ambalavanan, Namasivayam; Ansong, Charles

    2016-12-22

    Global proteomics approaches allow characterization of whole tissue lysates to an impressive depth. However, it is now increasingly recognized that to better understand the complexity of multicellular organisms, global protein profiling of specific spatially defined regions/substructures of tissues (i.e. spatially-resolved proteomics) is essential. Laser capture microdissection (LCM) enables microscopic isolation of defined regions of tissues preserving crucial spatial information. However, current proteomics workflows entail several manual sample preparation steps and are challenged by the microscopic mass-limited samples generated by LCM, and that impact measurement robustness, quantification, and throughput. Here, we coupled LCM with a fully automated sample preparation workflow that with a single manual step allows: protein extraction, tryptic digestion, peptide cleanup and LC-MS/MS analysis of proteomes from microdissected tissues. Benchmarking against the current state of the art in ultrasensitive global proteomic analysis, our approach demonstrated significant improvements in quantification and throughput. Using our LCM-SNaPP proteomics approach, we characterized to a depth of more than 3,400 proteins, the ontogeny of protein changes during normal lung development in laser capture microdissected alveolar tissue containing ~4,000 cells per sample. Importantly, the data revealed quantitative changes for 350 low abundance transcription factors and signaling molecules, confirming earlier transcript-level observations and defining seven modules of coordinated transcription factor/signaling molecule expression patterns, suggesting that a complex network of temporal regulatory control directs normal lung development with epigenetic regulation fine-tuning pre-natal developmental processes. Our LCM-proteomics approach facilitates efficient, spatially-resolved, ultrasensitive global proteomics analyses in high-throughput that will be enabling for several clinical and

  1. MicroRNA Stability in FFPE Tissue Samples: Dependence on GC Content.

    Science.gov (United States)

    Kakimoto, Yu; Tanaka, Masayuki; Kamiguchi, Hiroshi; Ochiai, Eriko; Osawa, Motoki

    2016-01-01

    MicroRNAs (miRNAs) are small non-coding RNAs responsible for fine-tuning of gene expression at post-transcriptional level. The alterations in miRNA expression levels profoundly affect human health and often lead to the development of severe diseases. Currently, high throughput analyses, such as microarray and deep sequencing, are performed in order to identify miRNA biomarkers, using archival patient tissue samples. MiRNAs are more robust than longer RNAs, and resistant to extreme temperatures, pH, and formalin-fixed paraffin-embedding (FFPE) process. Here, we have compared the stability of miRNAs in FFPE cardiac tissues using next-generation sequencing. The mode read length in FFPE samples was 11 nucleotides (nt), while that in the matched frozen samples was 22 nt. Although the read counts were increased 1.7-fold in FFPE samples, compared with those in the frozen samples, the average miRNA mapping rate decreased from 32.0% to 9.4%. These results indicate that, in addition to the fragmentation of longer RNAs, miRNAs are to some extent degraded in FFPE tissues as well. The expression profiles of total miRNAs in two groups were highly correlated (0.88 tissues instead of miR-1, which was predominant before fixation. Subsequent quantitative PCR (qPCR) analyses revealed that miRNAs with GC content of less than 40% are more degraded than GC-rich miRNAs (p<0.0001). We showed that deep sequencing data obtained using FFPE samples cannot be directly compared with that of fresh frozen samples. The combination of miRNA deep sequencing and other quantitative analyses, such as qPCR, may improve the utility of archival FFPE tissue samples.

  2. Screening of Viral Pathogens from Pediatric Ileal Tissue Samples after Vaccination

    Directory of Open Access Journals (Sweden)

    Laura Hewitson

    2014-01-01

    Full Text Available In 2010, researchers reported that the two US-licensed rotavirus vaccines contained DNA or DNA fragments from porcine circovirus (PCV. Although PCV, a common virus among pigs, is not thought to cause illness in humans, these findings raised several safety concerns. In this study, we sought to determine whether viruses, including PCV, could be detected in ileal tissue samples of children vaccinated with one of the two rotavirus vaccines. A broad spectrum, novel DNA detection technology, the Lawrence Livermore Microbial Detection Array (LLMDA, was utilized, and confirmation of viral pathogens using the polymerase chain reaction (PCR was conducted. The LLMDA technology was recently used to identify PCV from one rotavirus vaccine. Ileal tissue samples were analyzed from 21 subjects, aged 15–62 months. PCV was not detected in any ileal tissue samples by the LLMDA or PCR. LLMDA identified a human rotavirus A from one of the vaccinated subjects, which is likely due to a recent infection from a wild type rotavirus. LLMDA also identified human parechovirus, a common gastroenteritis viral infection, from two subjects. Additionally, LLMDA detected common gastrointestinal bacterial organisms from the Enterobacteriaceae, Bacteroidaceae, and Streptococcaceae families from several subjects. This study provides a survey of viral and bacterial pathogens from pediatric ileal samples, and may shed light on future studies to identify pathogen associations with pediatric vaccinations.

  3. Swine infectious agents in Tayassu pecari and Pecari tajacu tissue samples from Brazil.

    Science.gov (United States)

    de Castro, Alessandra Marnie Martins Gomes; Brombila, Talita; Bersano, Josete Garcia; Soares, Herbert Sousa; Silva, Sheila Oliveira de Souza; Minervino, Antonio Humberto Hamad; Ogata, Renato Akio; Gennari, Solange Maria; Richtzenhain, Leonardo Jose

    2014-04-01

    Peccaries and pigs, Tayassuidae and Suidae respectively, diverged approximately one million years ago from a common ancestor. Because these families share some pathogens, peccaries can act as reservoirs of infectious pathogens for domestic and wild swine. We evaluated the presence of swine infectious agents in the spleen and lung tissues of white-lipped peccaries (WLP; Tayassu pecari) and collared peccaries (CP; Pecari tajacu) in Brazil. Samples from 10 adult CP and three WLP, which had been hunted by locals or hit by motor vehicles, were obtained from two free-ranging Brazilian populations. The samples were tested by PCR for Mycoplasma hyopneumoniae, Bordetella bronchiseptica, Pasteurella multocida, porcine circovirus 2 (PCV2), Suid herpesvirus 1 (SuHV-1), and porcine parvovirus (PPV). Positive samples were sequenced. Both species were negative for PPV and B. bronchiseptica and positive for PCV2 and SuHV-1. The lungs of two animals were positive for M. hyopneumoniae and P. multocida. This report is the first demonstration of PCV2 and SuHV-1 swine viruses and of M. hyopneumoniae and P. multocida bacteria in peccaries. One factor contributing to this detection was access to tissue samples, which is uncommon. The role of these infectious agents in peccaries is unknown and further epidemiologic studies should be performed. This study identified several infectious agents in peccaries and highlighted the importance of the tissue type used to detect pathogens.

  4. Expression of cyclo-oxygenase-2 enzyme in the tissue samples of patients with various clinicopathological stages of oral leukoplakia and oral squamous cell carcinoma

    Directory of Open Access Journals (Sweden)

    Nelson Aruldoss

    2016-01-01

    Full Text Available Aim: The purpose of this study was to evaluate the expression of cyclo-oxygenase-2 (COX-2 enzyme in the tissue samples of patients with various clinicopathological stages of oral leukoplakia and oral squamous cell carcinoma (OSCC. Materials and Methods: The samples for the study were divided into 4 groups. Group A comprised 20 healthy individuals with no habits. Twenty healthy individuals with habitual tobacco usage and no oral lesions were included in Group B. Twenty cases of leukoplakia diagnosed clinically and histopathologically were included in Group C. Staging was done using the modified classification and staging system of oral leukoplakia. Twenty cases of OSCC diagnosed clinically and histopathologically were included in Group D. Immunohistochemical staining was done on these 80 samples (paraffin blocks for COX-2 expression by indirect method using polymer based Horseradish peroxidase system. Statistical analysis was performed using Kruskal-Wallis test and Spearman′s rank correlation test. Results: Significant and proportional increase of COX-2 staining was noted with the increase in the severity of dysplasia. Eighty percent of OSCC expressed COX-2, increasing in its intensity of staining with the decrease in differentiation. Seventy five percent of leukoplakia showed positive COX-2 expression. Only 15% of positive controls were COX-2 positive. No normal mucosa showed positive expression of COX-2. Conclusion: High expression of COX-2 is seen in advanced stages of leukoplakia and OSCC. Hence, COX-2 enzyme increases cell proliferation, promotes angiogenesis and inhibits immune surveillance in carcinogenesis; it can be an early detection marker in oral leukoplakia and a prognostic marker of OSCC.

  5. Quantitative PIXE and PIGME analysis of milligram samples of biomineralized tissue in the limpet Patella vulgata

    Science.gov (United States)

    Kim, K.-S.; Webb, J.; Macey, D. J.; Cohen, D. D.

    1987-03-01

    Procedures have been developed to determine, by thick target PIXE and PIGME, the quantitative elemental composition of biological samples with a mass of approximately 1 mg. Systems of particular interest are the biomineralized tissues of chitons and limpets, marine invertebrates of global distribution whose radula teeth and associated tissue contain, variously, inorganic components at different stages of mineralization, e.g. Fe, Ca, P, F, Si, Cu. For the biomineralized teeth and tissue in the limpet Patella vulgata the content of Fe, Ca and P increases rapidly at an early stage of mineralization, while the Si content increases somewhat later. In fully mineralized teeth, the Fe and Si contents are comparable. These data are compared with previous results (Trends Biochem. Sci. 10 (1985) 6) obtained using the Oxford scanning proton microprobe.

  6. CT-guided Irreversible Electroporation in an Acute Porcine Liver Model: Effect of Previous Transarterial Iodized Oil Tissue Marking on Technical Parameters, 3D Computed Tomographic Rendering of the Electroporation Zone, and Histopathology

    Energy Technology Data Exchange (ETDEWEB)

    Sommer, C. M., E-mail: christof.sommer@med.uni-heidelberg.de [University Hospital Heidelberg, Department of Diagnostic and Interventional Radiology (Germany); Fritz, S., E-mail: stefan.fritz@med.uni-heidelberg.de [University Hospital Heidelberg, Department of General Visceral and Transplantation Surgery (Germany); Vollherbst, D., E-mail: dominikvollherbst@web.de [University Hospital Heidelberg, Department of Diagnostic and Interventional Radiology (Germany); Zelzer, S., E-mail: s.zelzer@dkfz-heidelberg.de [German Cancer Research Center (dkfz), Medical and Biological Informatics (Germany); Wachter, M. F., E-mail: fredericwachter@googlemail.com; Bellemann, N., E-mail: nadine.bellemann@med.uni-heidelberg.de; Gockner, T., E-mail: theresa.gockner@med.uni-heidelberg.de; Mokry, T., E-mail: theresa.mokry@med.uni-heidelberg.de; Schmitz, A., E-mail: anne.schmitz@med.uni-heidelberg.de [University Hospital Heidelberg, Department of Diagnostic and Interventional Radiology (Germany); Aulmann, S., E-mail: sebastian.aulmann@mail.com [University Hospital Heidelberg, Department of General Pathology (Germany); Stampfl, U., E-mail: ulrike.stampfl@med.uni-heidelberg.de [University Hospital Heidelberg, Department of Diagnostic and Interventional Radiology (Germany); Pereira, P., E-mail: philippe.pereira@slk-kliniken.de [SLK Kliniken Heilbronn GmbH, Clinic for Radiology, Minimally-invasive Therapies and Nuclear Medicine (Germany); Kauczor, H. U., E-mail: hu.kauczor@med.uni-heidelberg.de [University Hospital Heidelberg, Department of Diagnostic and Interventional Radiology (Germany); Werner, J., E-mail: jens.werner@med.uni-heidelberg.de [University Hospital Heidelberg, Department of General Visceral and Transplantation Surgery (Germany); Radeleff, B. A., E-mail: boris.radeleff@med.uni-heidelberg.de [University Hospital Heidelberg, Department of Diagnostic and Interventional Radiology (Germany)

    2015-02-15

    PurposeTo evaluate the effect of previous transarterial iodized oil tissue marking (ITM) on technical parameters, three-dimensional (3D) computed tomographic (CT) rendering of the electroporation zone, and histopathology after CT-guided irreversible electroporation (IRE) in an acute porcine liver model as a potential strategy to improve IRE performance.MethodsAfter Ethics Committee approval was obtained, in five landrace pigs, two IREs of the right and left liver (RL and LL) were performed under CT guidance with identical electroporation parameters. Before IRE, transarterial marking of the LL was performed with iodized oil. Nonenhanced and contrast-enhanced CT examinations followed. One hour after IRE, animals were killed and livers collected. Mean resulting voltage and amperage during IRE were assessed. For 3D CT rendering of the electroporation zone, parameters for size and shape were analyzed. Quantitative data were compared by the Mann–Whitney test. Histopathological differences were assessed.ResultsMean resulting voltage and amperage were 2,545.3 ± 66.0 V and 26.1 ± 1.8 A for RL, and 2,537.3 ± 69.0 V and 27.7 ± 1.8 A for LL without significant differences. Short axis, volume, and sphericity index were 16.5 ± 4.4 mm, 8.6 ± 3.2 cm{sup 3}, and 1.7 ± 0.3 for RL, and 18.2 ± 3.4 mm, 9.8 ± 3.8 cm{sup 3}, and 1.7 ± 0.3 for LL without significant differences. For RL and LL, the electroporation zone consisted of severely widened hepatic sinusoids containing erythrocytes and showed homogeneous apoptosis. For LL, iodized oil could be detected in the center and at the rim of the electroporation zone.ConclusionThere is no adverse effect of previous ITM on technical parameters, 3D CT rendering of the electroporation zone, and histopathology after CT-guided IRE of the liver.

  7. Detection of Flavobacterium psychrophilum from fish tissue and water samples by PCR amplification

    DEFF Research Database (Denmark)

    Wiklund, T.; Madsen, Lone; Bruun, Morten Sichlau

    2000-01-01

    Rainbow trout fry syndrome and cold-water disease, caused by Flavobacterium psychrophilum, are important diseases in farmed salmonids. Some of the presently available techniques for the detection of Fl. psychrophilum are either time consuming or lack sufficient sensitivity. In the present...... investigation, the possible detection of Fl. psychrophilum from fish tissue and water samples was examined using nested PCR with DNA probes against a sequence of the 16S rRNA genes. The DNA was extracted using Chelex(R) 100 chelating resin. The primers, which were tested against strains isolated from diseased...... to be more sensitive than agar cultivation of tissue samples from the brain of rainbow trout injected with Fl. psychrophilum. In non-sterile fresh water seeded with Fl. psychrophilum the detection limit of the PCR- assay was 1.7 cfu in the PCR tube, corresponding to 110 cfu ml(-1) water. The PCR...

  8. Effects of sample preparation on the optical properties of breast tissue

    Science.gov (United States)

    Marks, Fay A.

    1996-04-01

    The optical properties of biological tissue should be determined in vivo whenever possible. However, for those instances when in vivo studies are impractical, too expensive or inappropriate, and when blood flow is not an issue, the ability to perform in vitro studies then becomes invaluable. Optical absorption spectroscopy shows that it may be possible to obtain meaningful information about the optical properties of human breast tissue from in vitro samples if strict preparation and measuring protocols are used. That a strict protocol for storing and handling tissue is critical can be seen from our observations of changes in the optical absorption spectra that occur in response to formalin fixation, the passage of time, application of stains and dyes, and storage in growth medium of the excised tissue. In vivo optical absorption spectroscopy measurements have been made on human breast cancer xenografts and compared with in vitro measurements on breast biopsies prepared according to precise collection and treatment protocols. There is a 'window of opportunity' before time dependent changes in the UV optical absorption spectra of the excised tissue specimens occur. This time window of opportunity widens at longer wavelengths with the least changes occurring in the optical spectra in the NIR.

  9. Identification of organ tissue types and skin from forensic samples by microRNA expression analysis.

    Science.gov (United States)

    Sauer, Eva; Extra, Antje; Cachée, Philipp; Courts, Cornelius

    2017-05-01

    The identification of organ tissues in traces recovered from scenes and objects with regard to violent crimes involving serious injuries can be of considerable relevance in forensic investigations. Molecular genetic approaches are provably superior to histological and immunological assays in characterizing organ tissues, and micro-RNAs (miRNAs), due to their cell type specific expression patterns and stability against degradation, emerged as a promising molecular species for forensic analyses, with a range of tried and tested indicative markers. Thus, herein we present the first miRNA based approach for the forensic identification of organ tissues. Using quantitative PCR employing an empirically derived strategy for data normalization and unbiased statistical decision making, we assessed the differential expression of 15 preselected miRNAs in tissues of brain, kidney, lung, liver, heart muscle, skeletal muscle and skin. We show that not only can miRNA expression profiling be used to reliably differentiate between organ tissues but also that this method, which is compatible with and complementary to forensic DNA analysis, is applicable to realistic forensic samples e.g. mixtures, aged and degraded material as well as traces generated by mock stabbings and experimental shootings at ballistic models.

  10. Transgenic zebrafish reveal tissue-specific differences in estrogen signaling in response to environmental water samples

    Science.gov (United States)

    Gorelick, Daniel A.; Iwanowicz, Luke R.; Hung, Alice L.; Blazer, Vicki; Halpern, Marnie E.

    2014-01-01

    Background: Environmental endocrine disruptors (EED) are exogenous chemicals that mimic endogenous hormones, such as estrogens. Previous studies using a zebrafish transgenic reporter demonstrated that the EEDs bisphenol A and genistein preferentially activate estrogen receptors (ER) in the larval heart compared to the liver. However, it was not known whether the transgenic zebrafish reporter was sensitive enough to detect estrogens from environmental samples, whether environmental estrogens would exhibit similar tissue-specific effects as BPA and genistein or why some compounds preferentially target receptors in the heart. Methods: We tested surface water samples using a transgenic zebrafish reporter with tandem estrogen response elements driving green fluorescent protein expression (5xERE:GFP). Reporter activation was colocalized with tissue-specific expression of estrogen receptor genes by RNA in situ hybridization. Results: Selective patterns of ER activation were observed in transgenic fish exposed to river water samples from the Mid-Atlantic United States, with several samples preferentially activating receptors in embryonic and larval heart valves. We discovered that tissue-specificity in ER activation is due to differences in the expression of estrogen receptor subtypes. ERα is expressed in developing heart valves but not in the liver, whereas ERβ2 has the opposite profile. Accordingly, subtype-specific ER agonists activate the reporter in either the heart valves or the liver. Conclusion: The use of 5xERE:GFP transgenic zebrafish has revealed an unexpected tissue-specific difference in the response to environmentally relevant estrogenic compounds. Exposure to estrogenic EEDs in utero is associated with adverse health effects, with the potentially unanticipated consequence of targeting developing heart valves.

  11. A single lysis solution for the analysis of tissue samples by different proteomic technologies

    DEFF Research Database (Denmark)

    Gromov, P.; Celis, J.E.; Gromova, I.

    2008-01-01

    Cancer, being a major healthcare concern worldwide, is one of the main targets for the application of emerging proteomic technologies and these tools promise to revolutionize the way cancer will be diagnosed and treated in the near future. Today, as a result of the unprecedented advances that have...... dissease, is driving scientists to increasingly use clinically relevant samples for biomarker and target discovery. Tissues are heterogeneous and as a result optimization of sample preparation is critical for generating accurate, representative, and highly reproducible quantitative data. Although a large...

  12. The privacy of Tutankhamen--utilising the genetic information in stored tissue samples.

    Science.gov (United States)

    Holm, S

    2001-09-01

    Recent technical developments in genetic testing has led to a situation where the DNA in previously stored tissue samples can be extracted and used for genetic analysis. This raises the question of how to decide whether a specific use of such samples should be allowed. Using the genetic testing of ancient DNA in general, and the DNA of the pharaoh Tutankhamen in particular as examples this paper analyses the question. It investigates whether ethical frameworks based on proxy consent, cultural affiliation, ownership, or the privacy rights of the dead are appropriate and justifiable in this context. The conclusion is that frameworks based on proxy consent, cultural affiliation, and ownership are not very useful.

  13. Investigations on the radioimmunological determination of stilbenes in tissue samples from pigs

    Energy Technology Data Exchange (ETDEWEB)

    Hoffmann, B.; Weiler, S.

    Based on the extraction procedures described for tissue of veal calves and following the introduction of reversed-phase-column chromatography as an alternative purification step, a radioimmunoassay is described for the determination of DES in pork-tissue. When using the DES-specific antiserum AS 254 and depending on the tissue examined, the lower limits of detection were between 29-69 pg. A mean enor of 10.8% (CV between 13-17%) for the ecovery added to tissue samples and a CV between 3.6 and 10.5% for the reproducibility demonstrate a good reliability. Testing the stilbene-specific antiserum AS 6139 by using /sup 3/H-DES as tracer revealed, the DES - and to a lesser extent also HEX - could be quantified with an acceptable degree of reliability when using the homologous RIA-system (use of DES and HEX resp. as calibration-standard), differently to DIEN; the possibility of a transformation of DIEN during the extraction into a compound exhibiting a higher cross reactivity is discussed. Application of the heterologous test system (quantification of the sample-stilbene by using each of the other two stilbenes for calibration) yielded the expected over- and underestimations. Furtheron, in respect to total stilbenes, i.e. not knowing whether DES, HEX or DIEN are present in the sample, it has been shown, that the assay - though qualitative - is highly sensitive (59-86 pg lower level of detection) when DES is used as tracer and for calibration purposes.

  14. Threshold-dependent sample sizes for selenium assessment with stream fish tissue

    Science.gov (United States)

    Hitt, Nathaniel P.; Smith, David R.

    2015-01-01

    Natural resource managers are developing assessments of selenium (Se) contamination in freshwater ecosystems based on fish tissue concentrations. We evaluated the effects of sample size (i.e., number of fish per site) on the probability of correctly detecting mean whole-body Se values above a range of potential management thresholds. We modeled Se concentrations as gamma distributions with shape and scale parameters fitting an empirical mean-to-variance relationship in data from southwestern West Virginia, USA (63 collections, 382 individuals). We used parametric bootstrapping techniques to calculate statistical power as the probability of detecting true mean concentrations up to 3 mg Se/kg above management thresholds ranging from 4 to 8 mg Se/kg. Sample sizes required to achieve 80% power varied as a function of management thresholds and Type I error tolerance (α). Higher thresholds required more samples than lower thresholds because populations were more heterogeneous at higher mean Se levels. For instance, to assess a management threshold of 4 mg Se/kg, a sample of eight fish could detect an increase of approximately 1 mg Se/kg with 80% power (given α = 0.05), but this sample size would be unable to detect such an increase from a management threshold of 8 mg Se/kg with more than a coin-flip probability. Increasing α decreased sample size requirements to detect above-threshold mean Se concentrations with 80% power. For instance, at an α-level of 0.05, an 8-fish sample could detect an increase of approximately 2 units above a threshold of 8 mg Se/kg with 80% power, but when α was relaxed to 0.2, this sample size was more sensitive to increasing mean Se concentrations, allowing detection of an increase of approximately 1.2 units with equivalent power. Combining individuals into 2- and 4-fish composite samples for laboratory analysis did not decrease power because the reduced number of laboratory samples was compensated for by increased

  15. Evaluation of sample preparation methods and optimization of nickel determination in vegetable tissues

    Directory of Open Access Journals (Sweden)

    Rodrigo Fernando dos Santos Salazar

    2011-02-01

    Full Text Available Nickel, although essential to plants, may be toxic to plants and animals. It is mainly assimilated by food ingestion. However, information about the average levels of elements (including Ni in edible vegetables from different regions is still scarce in Brazil. The objectives of this study were to: (a evaluate and optimize a method for preparation of vegetable tissue samples for Ni determination; (b optimize the analytical procedures for determination by Flame Atomic Absorption Spectrometry (FAAS and by Electrothermal Atomic Absorption (ETAAS in vegetable samples and (c determine the Ni concentration in vegetables consumed in the cities of Lorena and Taubaté in the Vale do Paraíba, State of São Paulo, Brazil. By means of the analytical technique for determination by ETAAS or FAAS, the results were validated by the test of analyte addition and recovery. The most viable method tested for quantification of this element was HClO4-HNO3 wet digestion. All samples but carrot tissue collected in Lorena contained Ni levels above the permitted by the Brazilian Ministry of Health. The most disturbing results, requiring more detailed studies, were the Ni concentrations measured in carrot samples from Taubaté, where levels were five times higher than permitted by Brazilian regulations.

  16. Application of a battery of biomarkers in mussel digestive gland to assess long-term effects of the Prestige oil spill in Galicia and Bay of Biscay: tissue-level biomarkers and histopathology.

    Science.gov (United States)

    Garmendia, Larraitz; Soto, Manu; Vicario, Unai; Kim, Yungkul; Cajaraville, Miren P; Marigómez, Ionan

    2011-04-01

    In order to assess the biological effects of the Prestige oil spill (POS), mussels, Mytilus galloprovincialis, were collected in 22 localities along the North coast of the Iberian Peninsula over 3 years (April 2003-April 2006). Different tissue-level biomarkers including cell type composition (volume density of basophilic cells, Vv(BAS)) in digestive gland epithelium, structural changes of digestive alveoli (mean luminal radius to mean epithelial thickness, MLR/MET) and histopathological alterations (prevalence and intensity) of the digestive gland were analysed. Severe alterations in the general condition of the digestive gland tissue were observed all over the study area up to 2004-2005. High Vv(BAS) values were recorded mainly in Galicia but also to a lesser extent in the Bay of Biscay in 2003-2004. Atrophy of the digestive alveoli, measured in terms of MLR/MET, was detected all along the studied area up to 2006. Inflammatory responses cannot be related to pollution due to the POS: (a) prevalence and intensity of focal hemocytic infiltration were higher in the Bay of Biscay than in Galicia but they did not show a clear temporal trend; (b) high intensities of brown cell aggregates were only sporadically recorded; and (c) granulocytomas were more frequently recorded in the Bay of Biscay than in Galicia and especially in localities (i.e. Arrigunaga) subjected to chronic pollution. Likewise, Marteilia, trematodes, intracellular ciliates, unidentified eosinophilic bodies, R/CLO and Mytilicola did not follow any recognisable pattern that could be associated to the POS. In contrast, high Nematopsis intensities recorded in several localities in 2003 might suggest some response of local interest after the POS (i.e., in combination with particular factors/conditions). More data at a regional scale are needed before histopathology may provide a reliable ecosystem health assessment but the present results suggest that the approach is worthwhile. Overall, although Vv

  17. Impact of freezing delay time on tissue samples for metabolomic studies

    Directory of Open Access Journals (Sweden)

    Tonje Husby Haukaas

    2016-01-01

    Full Text Available Introduction: Metabolic profiling of intact tumor tissue by high resolution magic angle spinning (HR MAS MR spectroscopy (MRS provides important biological information possibly useful for clinical diagnosis and development of novel treatment strategies. However, generation of high-quality data requires that sample handling from surgical resection until analysis is performed using systematically validated procedures. In this study, we investigated the effect of post-surgical freezing delay time on global metabolic profiles and stability of individual metabolites in intact tumor tissue.Materials and methods: Tumor tissue samples collected from two patient derived breast cancer xenograft models (n=3 for each model were divided into pieces that were snap-frozen in liquid nitrogen at 0, 15, 30, 60, 90, and 120 minutes after surgical removal. In addition, one sample was analysed immediately, representing the metabolic profile of fresh tissue exposed neither to liquid nitrogen nor to room temperature. We also evaluated the metabolic effect of prolonged spinning during the HR MAS experiments in biopsies from breast cancer patiens (n=14. All samples were analyzed by proton HR MAS MRS on a Bruker Avance DRX600 spectrometer, and changes in metabolic profiles were evaluated using multivariate analysis and linear mixed modeling (LMM. Results: Multivariate analysis showed that the metabolic differences between the two breast cancer models were more prominent than variation caused by freezing delay time. No significant changes in levels of individual metabolites were observed in samples frozen within 30 minutes of resection. After this time point, levels of choline increased whereas ascorbate, creatine and glutathione (GS levels decreased. Freezing had a significant effect on several metabolites, but is an essential procedure for research and biobank purposes. Furthermore, four metabolites (glucose, glycine, glycerophosphocholine and choline were affected by

  18. Learning histopathological patterns

    Directory of Open Access Journals (Sweden)

    Andreas Kårsnäs

    2011-01-01

    Full Text Available Aims: The aim was to demonstrate a method for automated image analysis of immunohistochemically stained tissue samples for extracting features that correlate with patient disease. We address the problem of quantifying tumor tissue and segmenting and counting cell nuclei. Materials and Methods: Our method utilizes a flexible segmentation method based on sparse coding trained from representative image samples. Nuclei counting is based on a nucleus model that takes size, shape, and nucleus probability into account. Nuclei clustering and overlays are resolved using a gray-weighted distance transform. We obtain a probability measure for pixels belonging to a nucleus from our segmentation procedure. Experiments are carried out on two sets of immunohistochemically stained images - one set based on the estrogen receptor (ER and the other on antigen KI-67. For the nuclei separation we have selected 207 ER image samples from 58 tissue micro array-cores corresponding to 58 patients and 136 KI-67 image samples also from 58 cores. The images are hand-annotated by marking the center position of each nucleus. For the ER data we have a total of 1006 nuclei and for the KI-67 we have 796 nuclei. Segmentation performance was evaluated in terms of missing nuclei, falsely detected nuclei, and multiple detections. The proposed method is compared to state-of-the-art Bayesian classification. Statistical analysis used: The performance of the proposed method and a state-of-the-art algorithm including variations thereof is compared using the Wilcoxon rank sum test. Results: For both the ER experiment and the KI-67 experiment the proposed method exhibits lower error rates than the state-of-the-art method. Total error rates were 4.8 % and 7.7 % in the two experiments, corresponding to an average of 0.23 and 0.45 errors per image, respectively. The Wilcoxon rank sum tests show statistically significant improvements over the state-of-the-art method. Conclusions: We have

  19. Histopathological changes in the livers and kidneys of fish in Sariyar Reservoir, Turkey.

    Science.gov (United States)

    Ayas, Zafer; Ekmekci, Guler; Ozmen, Murat; Yerli, Sedat V

    2007-03-01

    In this study, a total of 180 fish specimens (wels: Silurus glanis-60; common carp: Cyprinus carpio-60; bleak: Alburnus escherichii-60) of ages between one and two were caught at three different stations in Sariyar Reservoir. The histological changes in the livers and kidneys of three different species of fish were detected microscopically and evaluated with quantitative analyses. Also, organochlorine pesticide residues (OCP) have also been determined in the water and sediment samples and in the adipose tissues of fish caught in these stations. Results show that the reservoir was polluted by different kinds of OCP compounds and these chemicals have accumulated in the fish tissues. As a result of these analyses, histopathological changes were observed in the livers and kidneys of fish specimens, such as mononuclear cell infiltration, congestion and nuclear picnosis. Also intra-cytoplasmic cholestasis in their livers and tubular degenerations in the kidneys were observed. The incidences of the histopathological changes in wels and carps were found to be higher than bleak. Furthermore, histopathological changes in fish samples caught from Usakbuku were much more than the samples caught from other stations (Sariyar and Nallihan Bird Paradise Stations). In this study the possible reasons of histopathological changes were evaluated with respect to different fish species and localities and also the findings were evaluated in relation to OCP contamination.

  20. Implementation of immunohistochemistry on frozen ear notch tissue samples in diagnosis of bovine viral diarrhea virus in persistently infected cattle

    Directory of Open Access Journals (Sweden)

    Bedeković Tomislav

    2011-12-01

    Full Text Available Abstract Background Bovine viral diarrhea is a contagious disease of domestic and wild ruminants and one of the most economically important diseases in cattle. Bovine viral diarrhea virus belongs to the genus Pestivirus, within the family Flaviviridae. The identification and elimination of the persistently infected animals from herds is the initial step in the control and eradication programs. It is therefore necessary to have reliable methods for diagnosis of bovine viral diarrhea virus. One of those methods is immunohistochemistry. Immunohistochemistry on formalin fixed, paraffin embedded tissue is a routine technique in diagnosis of persistently infected cattle from ear notch tissue samples. However, such technique is inappropriate due to complicated tissue fixation process and it requires more days for preparation. On the contrary, immunohistochemistry on frozen tissue was usually applied on organs from dead animals. In this paper, for the first time, the imunohistochemistry on frozen ear notch tissue samples was described. Findings Seventeen ear notch tissue samples were obtained during the period 2008-2009 from persistently infected cattle. Samples were fixed in liquid nitrogen and stored on -20°C until testing. Ear notch tissue samples from all persistently infected cattle showed positive results with good section quality and possibility to determinate type of infected cells. Conclusions Although the number of samples was limited, this study indicated that immunohistochemistry on formalin fixed paraffin embedded tissue can be successfully replaced with immunohistochemistry on frozen ear notch tissue samples in diagnosis of persistently infected cattle.

  1. Expression of Wnt pathway mediators in metaplasic tissue in animal model and clinical samples of tendinopathy.

    Science.gov (United States)

    Lui, Pauline Po Yee; Lee, Yuk Wa; Wong, Yin Mei; Zhang, Xiaoling; Dai, Kerong; Rolf, Christer Gustav

    2013-09-01

    Tissue metaplasia is observed in both ossified failed healing animal model and clinical samples of tendinopathy. The Wnt signalling pathway plays a vital role in pathological calcification. We hypothesized that the Wnt signalling pathway might contribute to tissue metaplasia and failed healing in tendinopathy. This study aimed to examine the spatial-temporal expression of Wnt pathway mediators in an ossified failed tendon healing animal model and clinical samples of tendinopathy. The effect of Wnt3a on the osteogenic differentiation of tendon-derived stem cells (TDSCs) was also examined. Ossified failed tendon healing was induced by the injection of collagenase into the patellar tendon of rats. At various times the tendons were harvested for immunohistochemical staining of Wnt3a, β-catenin, Lrp5 and Tcf1. Patellar tendon samples were obtained from 13 patients with patellar tendinopathy (11 unossified and 2 ossified) and 10 controls. Immunohistochemical staining of Wnt3a, β-catenin, Lrp5 and Tcf1 was similarly performed. Rat patellar TDSCs were treated with Wnt3a. The osteogenic differentiation of TDSCs was examined by ALP activity, alizarin red S staining and mRNA expression of osteogenic markers. There was increased expression of Wnt3a, β-catenin, Lrp5 and Tcf1 in the healing fibroblast-like cells, chondrocyte-like cells and ossified deposits in the animal model and in some clinical samples of tendinopathy. Wnt3a increased ALP activity, calcium nodule formation and expression of osteogenic markers in TDSCs. Activation of the Wnt signalling pathway and its effect on TDSCs might contribute to tissue metaplasia and failed healing in some cases of tendinopathy.

  2. Magnetic resonance imaging and histopathology in the femoral head

    Energy Technology Data Exchange (ETDEWEB)

    Takatori, Yoshio; Kamogawa, Morihide; Nakamura, Toshitaka; Ninomiya, Setsuo; Kokubo, Takashi

    1989-05-01

    To correlate the magnetic resonance (MR) images with the histopathological findings in the femoral head, the histopathology of 24 femoral heads, 15 with osteonecrosis, five with osteoarthritis and four with other hip disorders were subjected to preoperative MR imaging which demonstrated low intensity areas due to long T1 relaxation time in the femoral head. The MR signal was low where fibrovascular tissue, disintegrated fibrovascular tissue, amorphous necrotic material, bone, or cartilagenous tissue occupied the medullary space. From this study, it seems possible to predict the histopathologic changes in the femoral head using MR images. (author).

  3. Measurement of characteristic prompt gamma rays emitted from oxygen and carbon in tissue-equivalent samples during proton beam irradiation

    OpenAIRE

    Polf, Jerimy C.; Panthi, Rajesh; Mackin, Dennis S; McCleskey, Matt; Saastamoinen, Antti; Roeder, Brian T; Beddar, Sam

    2013-01-01

    The purpose of this work was to characterize how prompt gamma (PG) emission from tissue changes as a function of carbon and oxygen concentration, and to assess the feasibility of determining elemental concentration in tissues irradiated with proton beams. For this study, four tissue-equivalent water-sucrose samples with differing densities and concentrations of carbon, hydrogen, and oxygen were irradiated with a 48 MeV proton pencil beam. The PG spectrum emitted from each sample was measured ...

  4. The accuracy of extended histopathology to detect immunotoxic chemicals

    NARCIS (Netherlands)

    Germolec, D.R.; Kashon, M.; Nyska, A.; Kuper, C.F.; Portier, C.; Kommineni, C.; Johnson, K.A.; Luster, M.I.

    2004-01-01

    The accuracy of extended histopathology to detect immunotoxic chemicals in female B6C3F1 mice was evaluated under the auspices of the National Toxicology Program (NTP). A workgroup was formed consisting of four pathologists who conducted extended histopathological evaluation of lymphoid tissues

  5. Histopathological changes observed in the heart and gizzard of ...

    African Journals Online (AJOL)

    Administrator

    2006-10-02

    Oct 2, 2006 ... 2Department of Pharmacology, FPGMI, Lahore, Pakistan. 3Department of ... histopathological studies and effects of different levels were studied in these tissues. Histopathological .... translocation of the pyloric adjacent to the isthmus of the stomach produ- ... Chromium also causes toxicological effects on ...

  6. Evaluation of Chromosomal Disorders in Tissue and Blood Samples in Patients with Oral Squamous Cell Carcinoma

    Directory of Open Access Journals (Sweden)

    A. Parvaneroo

    2004-12-01

    Full Text Available Statement of Problem: Many studies have indicated that genetic disturbances are common findings in patients with Oral Squamous Cell Carcinoma (OSCC. Identification of these changes can be helpful in diagnostic procedures of these tumors.Purpose: The aim of this study was to appraise the chromosomal disorders in blood and tissue patients with OSCC.Methods and Materials: In this descriptive study, the study group consisted of all OSCC patients who were referred to the Faculty of Dentistry, Tehran University of Medical Sciences, Maxillofacial Surgery Clinic of Shariati Hospital, and Amir Aalam Hospital fromSeptember 2000 to November 2002. In order to study chromosomal disorders in the peripheral blood lymphocytes, 5 mL of blood was obtained from each patient In patients with the large lesion, a piece of involved tissue were obtained and cultured for 24 hours.This led to 29 blood samples and 16 tissue specimens and any relation between OSCC and age, sex, smoking and alcohol use were evaluated.Results: In this study, OSCC was more common in males than in females (3 to 5. 31% of our patients were smokers, and one had a history of alcoholic consumption. There was an increase in incidence of OSCC with age. In this study, all patients had numerical(aneuploidy, polyploidy and structural chromosomal disorders (double minute, fragment,breakage and dicentric. There was significant difference between blood and tissue chromosomal disorders (aneuploidy, polyploidy,breakage in OSCC patients.Conclusion: It can be concluded that chromosomes in patients with OSCC might show some genetic aberration and evaluation of involved tissue might be better way for determining this disorders.

  7. Transcriptional profiling of degraded RNA in cryopreserved and fixed tissue samples obtained at autopsy

    Directory of Open Access Journals (Sweden)

    Alhasan Samir

    2006-12-01

    Full Text Available Abstract Background Traditional multiplexed gene expression methods require well preserved, intact RNA. Such specimens are difficult to acquire in clinical practice where formalin fixation is the standard procedure for processing tissue. Even when special handling methods are used to obtain frozen tissue, there may be RNA degradation; for example autopsy samples where degradation occurs both pre-mortem and during the interval between death and cryopreservation. Although specimens with partially degraded RNA can be analyzed by qRT-PCR, these analyses can only be done individually or at low levels of multiplexing and are laborious and expensive to run for large numbers of RNA targets. Methods We evaluated the ability of the cDNA-mediated Annealing, Selection, extension, and Ligation (DASL assay to provide highly multiplexed analyses of cryopreserved and formalin fixed, paraffin embedded (FFPE tissues obtained at autopsy. Each assay provides data on 1536 targets, and can be performed on specimens with RNA fragments as small as 60 bp. Results The DASL performed accurately and consistently with cryopreserved RNA obtained at autopsy as well as with RNA extracted from formalin-fixed paraffin embedded tissue that had a cryopreserved mirror image specimen with high quality RNA. In FFPE tissue where the cryopreserved mirror image specimen was of low quality the assay performed reproducibly on some but not all specimens. Conclusion The DASL assay provides reproducible results from cryopreserved specimens and many FFPE specimens obtained at autopsy. Gene expression analyses of these specimens may be especially valuable for the study of non-cancer endpoints, where surgical specimens are rarely available.

  8. Mathematical (diagnostic algorithms in the digitization of oral histopathology: The new frontier in histopathological diagnosis

    Directory of Open Access Journals (Sweden)

    Abhishek Banerjee

    2015-01-01

    Full Text Available The technological progress in the digitalization of a complete histological glass slide has opened a new door in the tissue based diagnosis. Automated slide diagnosis can be made possible by the use of mathematical algorithms which are formulated by binary codes or values. These algorithms (diagnostic algorithms include both object based (object features, structures and pixel based (texture measures. The intra- and inter-observer errors inherent in the visual diagnosis of a histopathological slide are largely replaced by the use of diagnostic algorithms leading to a standardized and reproducible diagnosis. The present paper reviews the advances in digital histopathology especially related to the use of mathematical algorithms (diagnostic algorithms in the field of oral histopathology. The literature was reviewed for data relating to the use of algorithms utilized in the construction of computational software with special applications in oral histopathological diagnosis. The data were analyzed, and the types and end targets of the algorithms were tabulated. The advantages, specificities and reproducibility of the software, its shortcomings and its comparison with traditional methods of histopathological diagnosis were evaluated. Algorithms help in automated slide diagnosis by creating software with possible reduced errors and bias with a high degree of specificity, sensitivity, and reproducibility. Akin to the identification of thumbprints and faces, software for histopathological diagnosis will in the near future be an important part of the histopathological diagnosis.

  9. Formalin-induced fluorescence reveals cell shape and morphology in biological tissue samples.

    Directory of Open Access Journals (Sweden)

    Ulrich Leischner

    Full Text Available Ultramicroscopy is a powerful tool to reveal detailed three-dimensional structures of large microscopical objects. Using high magnification, we observed that formalin induces fluorescence more in extra-cellular space and stains cellular structures negatively, rendering cells as dark objects in front of a bright background. Here, we show this effect on a three-dimensional image stack of a hippocampus sample, focusing on the CA1 region. This method, called FIF-Ultramicroscopy, allows for the three-dimensional observation of cellular structures in various tissue types without complicated staining techniques.

  10. Persistent synthetic chlorinated hydrocarbons in albatross tissue samples from Midway Atoll

    Energy Technology Data Exchange (ETDEWEB)

    Jones, P.D.; Hannah, D.J.; Buckland, S.J. [ESR:Environmental, Lower Hutt (New Zealand)] [and others

    1996-10-01

    Anthropogenic organic contaminants have been found in even the most remote locations. To assess the global distribution and possible effects of such contaminants, the authors examined the tissues of two species of albatross collected from Midway Atoll in the central North Pacific Ocean. These birds have an extensive feeding range covering much of the subtropical and northern Pacific Ocean. Anthropogenic contaminants were found at relatively great concentrations in these birds. The sum of 19 polychlorinated biphenyl (PCB) congeners ranged from 177 ng/g wet weight in eggs to 2,750 ng/g wet weight in adult fat. Total toxic equivalents (TEQs) derived from polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) ranged from 17.2 to 297 pg/g wet weight in the same tissues, while the inclusion of TEQs from PCBs increased these values to 48.4 and 769 pg/g wet weight, respectively. While contaminant concentrations varied between species and tissues, the contaminant profile was relatively uniform. The profile of contaminants detected was unusual in that much of the TEQs was contributed by two pentachlorinated congeners (2,3,4,7,8-pentachlorinated dibenzo-p-dioxin), and the profiles of PCB congeners did not match known sources. When compared to other studies the concentrations detected in the Midway Atoll samples were near or above the thresholds known to cause adverse effects in other fish-eating bird species.

  11. Biomarker discovery in heterogeneous tissue samples -taking the in-silico deconfounding approach

    Directory of Open Access Journals (Sweden)

    Parida Shreemanta K

    2010-01-01

    Full Text Available Abstract Background For heterogeneous tissues, such as blood, measurements of gene expression are confounded by relative proportions of cell types involved. Conclusions have to rely on estimation of gene expression signals for homogeneous cell populations, e.g. by applying micro-dissection, fluorescence activated cell sorting, or in-silico deconfounding. We studied feasibility and validity of a non-negative matrix decomposition algorithm using experimental gene expression data for blood and sorted cells from the same donor samples. Our objective was to optimize the algorithm regarding detection of differentially expressed genes and to enable its use for classification in the difficult scenario of reversely regulated genes. This would be of importance for the identification of candidate biomarkers in heterogeneous tissues. Results Experimental data and simulation studies involving noise parameters estimated from these data revealed that for valid detection of differential gene expression, quantile normalization and use of non-log data are optimal. We demonstrate the feasibility of predicting proportions of constituting cell types from gene expression data of single samples, as a prerequisite for a deconfounding-based classification approach. Classification cross-validation errors with and without using deconfounding results are reported as well as sample-size dependencies. Implementation of the algorithm, simulation and analysis scripts are available. Conclusions The deconfounding algorithm without decorrelation using quantile normalization on non-log data is proposed for biomarkers that are difficult to detect, and for cases where confounding by varying proportions of cell types is the suspected reason. In this case, a deconfounding ranking approach can be used as a powerful alternative to, or complement of, other statistical learning approaches to define candidate biomarkers for molecular diagnosis and prediction in biomedicine, in

  12. Histopathology of Lyme arthritis in LSH hamsters

    Energy Technology Data Exchange (ETDEWEB)

    Hejka, A.; Schmitz, J.L.; England, D.M.; Callister, S.M.; Schell, R.F.

    1989-05-01

    The authors studied the histopathologic evolution of arthritis in nonirradiated and irradiated hamsters infected with Borrelia burgdorferi. Nonirradiated hamsters injected in the hind paws with B. burgdorferi developed an acute inflammatory reaction involving the synovium, periarticular soft tissues, and dermis. This acute inflammatory reaction was short-lived and was replaced by a mild chronic synovitis as the number of detectable spirochetes in the synovium, periarticular soft tissues, and perineurovascular areas diminished. Exposing hamsters to radiation before inoculation with B. burgdorferi exacerbated and prolonged the acute inflammatory phase. Spirochetes also persisted longer in the periarticular soft tissues. A major histopathologic finding was destructive and erosive bone changes of the hind paws, which resulted in deformation of the joints. These studies should be helpful in defining the immune mechanism participating in the onset, progression, and resolution of Lyme arthritis.

  13. A percutaneous needle biopsy technique for sampling the supraclavicular brown adipose tissue depot of humans.

    Science.gov (United States)

    Chondronikola, M; Annamalai, P; Chao, T; Porter, C; Saraf, M K; Cesani, F; Sidossis, L S

    2015-10-01

    Brown adipose tissue (BAT) has been proposed as a potential target tissue against obesity and its related metabolic complications. Although the molecular and functional characteristics of BAT have been intensively studied in rodents, only a few studies have used human BAT specimens due to the difficulty of sampling human BAT deposits. We established a novel positron emission tomography and computed tomography-guided Bergström needle biopsy technique to acquire human BAT specimens from the supraclavicular area in human subjects. Forty-three biopsies were performed on 23 participants. The procedure was tolerated well by the majority of participants. No major complications were noted. Numbness (9.6%) and hematoma (2.3%) were the two minor complications noted, which fully resolved. Thus, the proposed biopsy technique can be considered safe with only minimal risk of adverse events. Adoption of the proposed method is expected to increase the sampling of the supraclavicular BAT depot for research purposes so as to augment the scientific knowledge of the biology of human BAT.

  14. Evaluation of convenient pretreatment protocols for RNA virus metagenomics in serum and tissue samples.

    Science.gov (United States)

    Rosseel, Toon; Ozhelvaci, Orkun; Freimanis, Graham; Van Borm, Steven

    2015-09-15

    Viral metagenomic approaches are increasingly being used for viral discovery. Various strategies are applied to enrich viral sequences, but there is often a lack of knowledge about their effective influence on the viral discovery sensitivity. We evaluate some convenient and widely used approaches for RNA virus discovery in clinical samples in order to reveal their sensitivity and potential bias introduced by the enrichment or amplifications steps. An RNA virus was artificially spiked at a fixed titer in serum and lung tissue, respectively, low and high nucleic acid content matrices. For serum, a simple DNase treatment on the RNA extract gave the maximum gain in proportion of viral sequences (83×), and a subsequent ribosomal RNA removal nearly doubled once more the proportion of viral sequences. For lung tissue, a ribosomal RNA depletion step on the RNA extract had the biggest gain in proportion of viral sequences (32×). We show also that direct sequencing of cDNA is recommended above an extra random PCR amplification step, and a that the virion enrichment strategy (filtration and nuclease treatment) has a beneficial effect for sequencing-based virus discovery. Our findings provide sample-dependent guidelines for targeted virus discovery strategies.

  15. Histopathologic approaches to detect changes indicative of immunotoxicity

    NARCIS (Netherlands)

    Kuper, C.F.; Harleman, J.H.; Richter-Reichelm, H.B.; Vos, J.G.

    2000-01-01

    Toxicologic pathology is crucial in the identification and characterization of health effects following exposure to xenobiotics, mainly in toxicity experiments in rodents. Regarding regulatory toxicology, histopathology of lymphoid organs and tissues is a cornerstone in the identification of immunot

  16. KRAS mutation: comparison of testing methods and tissue sampling techniques in colon cancer.

    Science.gov (United States)

    Franklin, Wilbur A; Haney, Jerry; Sugita, Michio; Bemis, Lynne; Jimeno, Antonio; Messersmith, Wells A

    2010-01-01

    Treatment of colon carcinoma with the anti-epidermal growth factor receptor antibody Cetuximab is reported to be ineffective in KRAS-mutant tumors. Mutation testing techniques have therefore become an urgent concern. We have compared three methods for detecting KRAS mutations in 59 cases of colon carcinoma: 1) high resolution melting, 2) the amplification refractory mutation system using a bifunctional self-probing primer (ARMS/Scorpion, ARMS/S), and 3) direct sequencing. We also evaluated the effects of the methods of sectioning and coring of paraffin blocks to obtain tumor DNA on assay sensitivity and specificity. The most sensitive and specific combination of block sampling and mutational analysis was ARMS/S performed on DNA derived from 1-mm paraffin cores. This combination of tissue sampling and testing method detected KRAS mutations in 46% of colon tumors. Four samples were positive by ARMS/S, but initially negative by direct sequencing. Cloned DNA samples were retested by direct sequencing, and in all four cases KRAS mutations were identified in the DNA. In six cases, high resolution melting abnormalities could not be confirmed as specific mutations either by ARMS/S or direct sequencing. We conclude that coring of the paraffin blocks and testing by ARMS/S is a sensitive, specific, and efficient method for KRAS testing.

  17. Quality and safety aspects in histopathology laboratory.

    Science.gov (United States)

    Adyanthaya, Soniya; Jose, Maji

    2013-09-01

    Histopathology is an art of analyzing and interpreting the shapes, sizes and architectural patterns of cells and tissues within a given specific clinical background and a science by which the image is placed in the context of knowledge of pathobiology, to arrive at an accurate diagnosis. To function effectively and safely, all the procedures and activities of histopathology laboratory should be evaluated and monitored accurately. In histopathology laboratory, the concept of quality control is applicable to pre-analytical, analytical and post-analytical activities. Ensuring safety of working personnel as well as environment is also highly important. Safety issues that may come up in a histopathology lab are primarily those related to potentially hazardous chemicals, biohazardous materials, accidents linked to the equipment and instrumentation employed and general risks from electrical and fire hazards. This article discusses quality management system which can ensure quality performance in histopathology laboratory. The hazards in pathology laboratories and practical safety measures aimed at controlling the dangers are also discussed with the objective of promoting safety consciousness and the practice of laboratory safety.

  18. Assessment of bioburden on human and animal tissues: part 2--results of testing of human tissue and qualification of a composite sample for routine bioburden determination.

    Science.gov (United States)

    Kowalski, John B; Merritt, Karen; Gocke, David; Osborne, Joel

    2012-08-01

    A quantitative method was developed and validated to assess bioburden on tissue from human donors and to compare bioburden determination results to swab culture results from the same donor. An initial study with allograft tissue from 101 donors showed a wide range of bioburden levels; values from no colony-forming units (CFU) detected to >28,000 CFU were observed. Tissues from donors that had swab cultures negative for objectionable microorganisms generally had lower bioburden than tissues from donors where objectionable microorganisms were recovered by swab culturing. In a follow-up study with 1,445 donors, a wide range of bioburden levels was again observed on tissues from donors that were swab culture negative for objectionable microorganisms. Tissues from 885 (61%) of these donors had no recoverable bioburden (donors had recoverable bioburden which ranged from 1 to >24,000 CFU. Identification of bioburden isolates showed a diversity of genera and species. In compliance with the recent revision of the American Association of Tissue Banks K2.210 Standard, the quantitative bioburden determination method was validated with a composite tissue sample that contains bone and soft tissue sections tested together in one extraction vessel. A recovery efficiency of 68% was validated and the composite sample was shown to be representative of all of the tissues recovered from a donor. The use of the composite sample in conjunction with the quantitative bioburden determination method will facilitate an accurate assessment of the numbers and types of contaminating microorganisms on allografts prior to disinfection/sterilization. This information will ensure that disinfection/sterilization processes are properly validated and the capability of the overall allograft process is understood on a donor by donor basis.

  19. Liquid Microjunction Surface Sampling Probe Electrospray Mass Spectrometry for Detection of Drugs and Metabolites in Thin Tissue Sections

    Energy Technology Data Exchange (ETDEWEB)

    Van Berkel, Gary J [ORNL; Kertesz, Vilmos [ORNL; Koeplinger, Kenneth A. [Merck Research Laboratories; Vavek, Marissa [Merck Research Laboratories; Kong, Ah-Ng Tony [Rutgers University

    2008-01-01

    A self-aspirating, liquid micro-junction surface sampling probe/electrospray emitter mass spectrometry system was demonstrated for use in the direct analysis of spotted and dosed drugs and their metabolites in thin tissue sections. Proof-of-principle sampling and analysis directly from tissue without the need for sample preparation was demonstrated first by raster scanning a region on a section of rat liver onto which reserpine was spotted. The mass spectral signal from selected reaction monitoring was used to develop a chemical image of the spotted drug on the tissue. The probe was also used to selectively spot sample areas of sagittal whole mouse body tissue sections that had been dosed orally (90 mg/kg) with R,S-sulforaphane 3 hrs prior to sacrifice. Sulforaphane and its glutathione and N-acetyl cysteine conjugates were monitored with selected reaction monitoring and detected in the stomach and various other tissues from the dosed mouse. No signal for these species was observed in the tissue from a control mouse. The same dosed tissue section was used to illustrate the possibility of obtaining a line scan across the whole body section. In total these results illustrate the potential for rapid screening of the distribution of drugs and metabolites in tissue sections with the micro-liquid junction surface sampling probe/electrospray mass spectrometry approach.

  20. Temporomandibular joint disc; a proposed histopathological degeneration grading score system

    OpenAIRE

    2010-01-01

    Summary. Aims: The purpose of this study was to draw up a TMJ disc histopathological score that is a semiquantitative transcription of the entire spectrum of TMJ disc degenerative diseases related to changes in disc tissue, and then validate the proposed grading, in order to contribute to a standardized histopathological diagnosis. Methods: Sections from sixty two temporomandibular joint disc specimens affected by tissue degenerative changes and stained with Hematox...

  1. Reporting tumor molecular heterogeneity in histopathological diagnosis.

    Directory of Open Access Journals (Sweden)

    Andrea Mafficini

    Full Text Available Detection of molecular tumor heterogeneity has become of paramount importance with the advent of targeted therapies. Analysis for detection should be comprehensive, timely and based on routinely available tumor samples.To evaluate the diagnostic potential of targeted multigene next-generation sequencing (TM-NGS in characterizing gastrointestinal cancer molecular heterogeneity.35 gastrointestinal tract tumors, five of each intestinal type gastric carcinomas, pancreatic ductal adenocarcinomas, pancreatic intraductal papillary mucinous neoplasms, ampulla of Vater carcinomas, hepatocellular carcinomas, cholangiocarcinomas, pancreatic solid pseudopapillary tumors were assessed for mutations in 46 cancer-associated genes, using Ion Torrent semiconductor-based TM-NGS. One ampulla of Vater carcinoma cell line and one hepatic carcinosarcoma served to assess assay sensitivity. TP53, PIK3CA, KRAS, and BRAF mutations were validated by conventional Sanger sequencing.TM-NGS yielded overlapping results on matched fresh-frozen and formalin-fixed paraffin-embedded (FFPE tissues, with a mutation detection limit of 1% for fresh-frozen high molecular weight DNA and 2% for FFPE partially degraded DNA. At least one somatic mutation was observed in all tumors tested; multiple alterations were detected in 20/35 (57% tumors. Seven cancers displayed significant differences in allelic frequencies for distinct mutations, indicating the presence of intratumor molecular heterogeneity; this was confirmed on selected samples by immunohistochemistry of p53 and Smad4, showing concordance with mutational analysis.TM-NGS is able to detect and quantitate multiple gene alterations from limited amounts of DNA, moving one step closer to a next-generation histopathologic diagnosis that integrates morphologic, immunophenotypic, and multigene mutational analysis on routinely processed tissues, essential for personalized cancer therapy.

  2. Reporting Tumor Molecular Heterogeneity in Histopathological Diagnosis

    Science.gov (United States)

    Mafficini, Andrea; Amato, Eliana; Fassan, Matteo; Simbolo, Michele; Antonello, Davide; Vicentini, Caterina; Scardoni, Maria; Bersani, Samantha; Gottardi, Marisa; Rusev, Borislav; Malpeli, Giorgio; Corbo, Vincenzo; Barbi, Stefano; Sikora, Katarzyna O.; Lawlor, Rita T.; Tortora, Giampaolo; Scarpa, Aldo

    2014-01-01

    Background Detection of molecular tumor heterogeneity has become of paramount importance with the advent of targeted therapies. Analysis for detection should be comprehensive, timely and based on routinely available tumor samples. Aim To evaluate the diagnostic potential of targeted multigene next-generation sequencing (TM-NGS) in characterizing gastrointestinal cancer molecular heterogeneity. Methods 35 gastrointestinal tract tumors, five of each intestinal type gastric carcinomas, pancreatic ductal adenocarcinomas, pancreatic intraductal papillary mucinous neoplasms, ampulla of Vater carcinomas, hepatocellular carcinomas, cholangiocarcinomas, pancreatic solid pseudopapillary tumors were assessed for mutations in 46 cancer-associated genes, using Ion Torrent semiconductor-based TM-NGS. One ampulla of Vater carcinoma cell line and one hepatic carcinosarcoma served to assess assay sensitivity. TP53, PIK3CA, KRAS, and BRAF mutations were validated by conventional Sanger sequencing. Results TM-NGS yielded overlapping results on matched fresh-frozen and formalin-fixed paraffin-embedded (FFPE) tissues, with a mutation detection limit of 1% for fresh-frozen high molecular weight DNA and 2% for FFPE partially degraded DNA. At least one somatic mutation was observed in all tumors tested; multiple alterations were detected in 20/35 (57%) tumors. Seven cancers displayed significant differences in allelic frequencies for distinct mutations, indicating the presence of intratumor molecular heterogeneity; this was confirmed on selected samples by immunohistochemistry of p53 and Smad4, showing concordance with mutational analysis. Conclusions TM-NGS is able to detect and quantitate multiple gene alterations from limited amounts of DNA, moving one step closer to a next-generation histopathologic diagnosis that integrates morphologic, immunophenotypic, and multigene mutational analysis on routinely processed tissues, essential for personalized cancer therapy. PMID:25127237

  3. Histopathological and Serological Studies on Paratuberculosis in Cattle and Buffaloes

    Directory of Open Access Journals (Sweden)

    Arbab Sikandar*, AH Cheema1, M Younus2, A Aslam1, MA Zaman2 and T Rehman3

    2012-10-01

    Full Text Available Paratuberculosis (Johne’s diseases is responsible for massive economic losses to dairy industry, both in the industrially advanced as well as in the developing countries. To detect its occurrence in cattle and buffaloes locally, blood and tissue samples from clinically weak and grossly suspected slaughtered animals were collected from two abattoirs of Jhang, municipal area, Pakistan. Acid-fast smear staining, gross/histopathology and indirect ELISA were done for the detection of Mycobacterium avium subsp. paratuberculosis (MAP. Total 134 samples illustrating gross pathological lesions were collected, only 11.19% (cattle: 6.67%, buffaloes: 12.5% showed acid fast bacilli through smear staining and were taken as confirmed cases. Thickening of intestines alone was not a reliable indicator of Johne’s disease. Tissue sections from intestines and mesenteric lymph nodes from these acid fast positive animals were stained with hematoxylin & eosin (H&E and Ziehl Neelsen (ZN methods. Sum of (15/134 impression smear staining as well as (15/15 tissue sections of the intestines were found ZN positive, and only 6.7% of impression smears and 100% of tissue sections of mesenteric lymph nodes showed acid fast bacilli. Through ELISA, two cattle and five buffaloes (07/134 gave positive optical densities, while one cattle and seven buffaloes (08/134 were judged as doubtful. It is concluded that infection of MAP can be identified by histopathology and ELISA. The present study was the first record of paratuberculosis among the dairy animals slaughtered at Jhang abattoirs. The objective was to compare different methods for the diagnosis of Johne’s disease.

  4. Optimizing EUS-guided liver biopsy sampling: comprehensive assessment of needle types and tissue acquisition techniques.

    Science.gov (United States)

    Schulman, Allison R; Thompson, Christopher C; Odze, Robert; Chan, Walter W; Ryou, Marvin

    2017-02-01

    EUS-guided liver biopsy sampling using FNA and, more recently, fine-needle biopsy (FNB) needles has been reported with discrepant diagnostic accuracy, in part due to differences in methodology. We aimed to compare liver histologic yields of 4 EUS-based needles and 2 percutaneous needles to identify optimal number of needle passes and suction. Six needle types were tested on human cadaveric tissue: one 19G FNA needle, one existing 19G FNB needle, one novel 19G FNB needle, one 22G FNB needle, and two 18G percutaneous needles (18G1 and 18G2). Two needle excursion patterns (1 vs 3 fanning passes) were performed on all EUS needles. Primary outcome was number of portal tracts. Secondary outcomes were degree of fragmentation and specimen adequacy. Pairwise comparisons were performed using t tests, with a 2-sided P samplings (48 per needle type) were performed. The novel 19G FNB needle had significantly increased mean portal tracts compared with all needle types. The 22G FNB needle had significantly increased portal tracts compared with the 18G1 needle (3.8 vs 2.5, P sampling. Investigations are underway to determine whether these results can be replicated in a clinical setting. Copyright © 2017 American Society for Gastrointestinal Endoscopy. Published by Elsevier Inc. All rights reserved.

  5. Sources of technical variability in quantitative LC-MS proteomics: human brain tissue sample analysis.

    Science.gov (United States)

    Piehowski, Paul D; Petyuk, Vladislav A; Orton, Daniel J; Xie, Fang; Moore, Ronald J; Ramirez-Restrepo, Manuel; Engel, Anzhelika; Lieberman, Andrew P; Albin, Roger L; Camp, David G; Smith, Richard D; Myers, Amanda J

    2013-05-03

    To design a robust quantitative proteomics study, an understanding of both the inherent heterogeneity of the biological samples being studied as well as the technical variability of the proteomics methods and platform is needed. Additionally, accurately identifying the technical steps associated with the largest variability would provide valuable information for the improvement and design of future processing pipelines. We present an experimental strategy that allows for a detailed examination of the variability of the quantitative LC-MS proteomics measurements. By replicating analyses at different stages of processing, various technical components can be estimated and their individual contribution to technical variability can be dissected. This design can be easily adapted to other quantitative proteomics pipelines. Herein, we applied this methodology to our label-free workflow for the processing of human brain tissue. For this application, the pipeline was divided into four critical components: Tissue dissection and homogenization (extraction), protein denaturation followed by trypsin digestion and SPE cleanup (digestion), short-term run-to-run instrumental response fluctuation (instrumental variance), and long-term drift of the quantitative response of the LC-MS/MS platform over the 2 week period of continuous analysis (instrumental stability). From this analysis, we found the following contributions to variability: extraction (72%) > instrumental variance (16%) > instrumental stability (8.4%) > digestion (3.1%). Furthermore, the stability of the platform and its suitability for discovery proteomics studies is demonstrated.

  6. Accumulation levels and characteristics of some pesticides in human adipose tissue samples from Southeast China.

    Science.gov (United States)

    Wang, Na; Shi, Lili; Kong, Deyang; Cai, Daoji; Cao, Yanzhong; Liu, Yongming; Pang, Guofang; Yu, Rongbin

    2011-08-01

    This paper presents a comprehensive study of pesticide levels and bio-accumulation characteristics in human adipose tissues among residents of Southeast China. A large number of adipose samples (n=633) were selected for 58 pesticides and were analyzed by high sensitive Gas Chromatography-Tandem Mass Spectrometry (GC-MS/MS). The results showed that POPs pesticides were frequently detected, including 2,4'-DDD, 2,4'-DDE, 2,4'-DDT, 4,4'-DDD, 4,4'-DDE, 4,4'-DDT, α-HCH, β-HCH, γ-HCH, δ-HCH, hexachlorobenzene (HCB), and mirex. Other detected pesticide species were dicofol, methamidophos and chlordimeform, which have rarely been reported. Comparing to different countries, the concentrations of total DDT and HCH in these three Chinese southeastern sites were in the middle range, whereas the HCB and mirex were in the lower end. A significant correlation was observed between region as well as age and POPs pesticide levels. Some pesticide residue levels were also found significantly correlated to occupation. However, there was no significant correlation between gender and pesticides. Meanwhile, it is interesting to find that mortality of malignant tumors tends to associate with the pesticides levels in human adipose tissue. More importantly, the measured data presented in this study provide realistic information which is useful for assessing human exposure to pesticides in the general population of Southeast China. Copyright © 2011 Elsevier Ltd. All rights reserved.

  7. A Fast Carrier Chromatin Immunoprecipitation Method Applicable to Microdissected Tissue Samples

    Science.gov (United States)

    Hao, Haiping; Liu, Hester; Gonye, Gregory; Schwaber, James S.

    2008-01-01

    Transcriptional regulation studies of CNS neurons are complicated by both cellular diversity and plasticity. Microdissection of specific functionally related populations of neurons can greatly reduce these issues, but typically excludes the use of many technologies due to tissue requirements, such as Chromatin Immunoprecipitation (ChIP), a powerful tool for studying in vivo protein-DNA interactions. We have developed a fast carrier ChIP (Fast CChIP) method for analyzing specific in vivo transcription factor-DNA interactions in as little as 0.2 mm3 brain tissue. Using an antibody against phosphorylated cyclic-AMP response element binding (CREB) protein, we confirmed phospho-CREB (pCREB) binding at the c-fos gene promoter. Then we further demonstrated the applicability of Fast CChIP in determining hypertension-induced pCREB binding at the c-fos gene promoter in the rat nucleus tractus solitarius (NTS), confirming CREB’s role in mediating hypertension-induced c-fos expression. This method will be broadly applicable to individual brain nucleus and biopsy/surgical samples. PMID:18502516

  8. Sources of Technical Variability in Quantitative LC-MS Proteomics: Human Brain Tissue Sample Analysis.

    Energy Technology Data Exchange (ETDEWEB)

    Piehowski, Paul D.; Petyuk, Vladislav A.; Orton, Daniel J.; Xie, Fang; Moore, Ronald J.; Ramirez Restrepo, Manuel; Engel, Anzhelika; Lieberman, Andrew P.; Albin, Roger L.; Camp, David G.; Smith, Richard D.; Myers, Amanda J.

    2013-05-03

    To design a robust quantitative proteomics study, an understanding of both the inherent heterogeneity of the biological samples being studied as well as the technical variability of the proteomics methods and platform is needed. Additionally, accurately identifying the technical steps associated with the largest variability would provide valuable information for the improvement and design of future processing pipelines. We present an experimental strategy that allows for a detailed examination of the variability of the quantitative LC-MS proteomics measurements. By replicating analyses at different stages of processing, various technical components can be estimated and their individual contribution to technical variability can be dissected. This design can be easily adapted to other quantitative proteomics pipelines. Herein, we applied this methodology to our label-free workflow for the processing of human brain tissue. For this application, the pipeline was divided into four critical components: Tissue dissection and homogenization (extraction), protein denaturation followed by trypsin digestion and SPE clean-up (digestion), short-term run-to-run instrumental response fluctuation (instrumental variance), and long-term drift of the quantitative response of the LC-MS/MS platform over the 2 week period of continuous analysis (instrumental stability). From this analysis, we found the following contributions to variability: extraction (72%) >> instrumental variance (16%) > instrumental stability (8.4%) > digestion (3.1%). Furthermore, the stability of the platform and its’ suitability for discovery proteomics studies is demonstrated.

  9. MRI evaluation of the histopathological characteristic of limb soft-tissue aggressive fibromatosis%肢体软组织侵袭性纤维瘤病组织病理特征的 MRI 研究

    Institute of Scientific and Technical Information of China (English)

    江波; 杨献峰; 赖英荣; 潘碧涛; 单卉; 陈应明; 孟悛非

    2009-01-01

    目的:探讨应用MRI评估肢体软组织侵袭性纤维瘤病( AF )组织病理特征的意义。方法分析20例AF的MRI和组织病理学表现;比较AF瘤内不同MRI信号区的T1 WI、T2 WI信号及强化程度,观察瘤内不同MRI信号区的HE染色与Masson三色染色表现。对所获数据进行配对t检验。结果(1)AF起源于骨骼肌(19/20),边缘分叶(20/20),呈浸润性生长(20/20);(2)三维动态对比增强血管造影(3D-DCEMRA)显示瘤体边缘少量爪状新生动脉(7/7)和轻度肿瘤染色(7/7);(3)根据MRI表现,20例AF分为Ⅰ区、Ⅱ区2种结构。Ⅰ区,在T1 WI、T2 WI上为极低信号,增强扫描不强化;Ⅱ区, T1WI 上呈等信号或稍高信号,T2WI 上呈稍高信号,增强后显著强化;(4)Ⅰ区的T1WI、T2WI信号及强化程度分别为(0.10±0.02)、(0.24±0.03)和(5.22±0.42)%,Ⅱ区依次分别为(0.79±0.04)、(3.05±0.08)和(151.5±8.61)%,两者的T1WI信号(t=67.37)、T2WI信号(t=196.56)及强化程度之间(t=76.62)的差异均有统计学意义(P值均<0.01);(5)AF由成纤维细胞、纤维细胞和胶原纤维束组成。 Masson三色染色上,Ⅰ区呈蓝色,为成熟的胶原纤维束;Ⅱ区不染色,主要成分为成纤维细胞与纤维细胞。结论 AF的Ⅰ区、Ⅱ区结构在MRI上有明显特征,MRI能准确反映AF的组织病理与生物学行为特征。%Objective To assess the value of using MRI to evaluate the histopathological characteristic of limb soft-tissue aggressive fibromatosis ( AF ) .Methods The MRI findings and histopathological data of 20 patients with AF were obtained and analyzed. The difference between the different signal regions in AF were compared of signal intensity in T1-weighted images, T2-weighted images and degree of enhancement.The data were processed with paired t test

  10. Histopathology of chromoblastomycosis.

    Science.gov (United States)

    Uribe, F; Zuluaga, A I; Leon, W; Restrepo, A

    1989-01-01

    A study aimed at determining the histopathologic appearance of chromoblastomycotic lesions was undertaken. Biopsies from 26 patients with the disease were examined. It was found that 23 cases (88.46%) exhibited the organized mixed mycotic granuloma - OMMA -, a granuloma modified by the presence polymorphonuclear neutrophils - PMN's. An equal proportion of cases exhibited pseudoepitheliomatous hyperplasia, with the epithelium playing an important role in the transepidermic elimination of the fungus. Healing of the lesions took place by fibrosis which was observed in 21 (80.77%) patients; this type of reaction was more common in the deeper areas of the dermis.

  11. 不同程度OSAHS患者腭咽软组织病理形态学改变%Histopathologic and morphological changes of palatopharyngeal soft tissue in patients with mild, moderate, and severe obstructive sleep apnea hypopnea syndrome

    Institute of Scientific and Technical Information of China (English)

    杨怀安; 刘艾竹; 李赛楠; 何岩

    2012-01-01

    To observe the histopathologic and morphological changes of palatopharyngeal soft tissues in patients with different degrees of obstructive sleep apnea hypopnea syndrome(OSAHS) patients. Method; Thirty-eight male OSAHS patients were devided into 3 groups according to AHI, namely mild group ( n = 10) , moderate group(n=13) ,and severe group (n=15). The soft palate tissues with partial palatopharyngeal arch and palatoglossal arch tissues were obtained from surgery and processed with conventional paraffin embedding. The sections were stained by HE and observed under a light microscope. The histological quantitative changes of the specimens were measured by analyzing the constituent ratios of glandular tissue, fat tissue and interstitial elements. Statistical analysis was performed. Result;㎡ptical microscope showed thatdOO times) , as the aggravation of the OSAHS,the soft palate squamous epithelial cells are swollenand irregular,exhibiting hyperkeratosis,accompanied by liquefied degeneration of basal cell ; The mucous membrane and submucosal connective tissue contain a certain number of lymphocytes infiltration. The mucosa and submucosal layer of loose connective tissue contain inflammatory cells and a lot of fat vacuoles can be observed; The soft palate mucous acini have inconsistent and irregular shape, among which there are a certain amount of fat cells infiltration. Some mucous acini are replaced by serous acini with dark stained cytoplasm; The palatoglossal muscle and palatopharyngeus muscle fibers can't be identifiedwith disordered arrangement of structure, showing pleomorphic changes including swelling,atrophy and degeneration. Some of elastic fibers were disrupted and a lot of fat cells infiltration was observed. ㏕he constituent ratios of the three kinds of tissues in soft palate from different degrees of OSAHS patients show that quantitative changes of glandular tissue and interstitial elements among the mild, moderate and severe OSAHS group patients

  12. Histopathologic and immunohistochemical features of Hashimoto thyroiditis

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    H Kazem Amani

    2011-01-01

    Full Text Available Background: Intrathyroid lymphoid tissue is accrued in Hashimoto thyroiditis (HT. Histologically, this acquired lymphoid tissue bears a close resemblance to mucosa-associated lymphoid tissue (MALT and can evolve to lymphoma. Aim: To demonstrate the morphological, and immunohistochemical profiles of Hashimoto thyroiditis and to ascertain the importance of light chain restriction in distinguishing HT with extensive lymphoplasmacytoid infiltrate from MALT lymphoma. Materials and Methods: We studied histopathologically and immunohistochemically (CD20, CD3, Igk, Igl and cytokeratin 30 cases of HT for evaluation of the lymphoid infiltrate and the presence of lymphoepithelial lesions (LELs. Distinguishing between early thyroid lymphoma and HT was evaluated by light chain restriction. These findings were compared with two cases of primary thyroid lymphoma. Results: The histopathological findings were characteristic of HT. Immunohistochemistry confirmed inconspicuous, rare B-cell LELs as well as a prominent T-lymphocyte population. Testing for light chain restriction showed polyclonal population of plasma cells. The cases of MALT lymphoma had distinct destructive lymphoepithelial lesions, B-cell immunophenotyping and showed kappa light chain restriction in the plasmacytoid population. Conclusions: Hashimoto thyroiditis differs both histopathologically and immunohistochemically from thyroid lymphoma. In suspicious cases, immunohistochemistry could be helpful in reaching a definitive diagnosis.

  13. Detection of Cryptococcus neoformans DNA in Tissue Samples by Nested and Real-Time PCR Assays

    Science.gov (United States)

    Bialek, Ralf; Weiss, Michael; Bekure-Nemariam, Kubrom; Najvar, Laura K.; Alberdi, Maria B.; Graybill, John R.; Reischl, Udo

    2002-01-01

    Two PCR protocols targeting the 18S rRNA gene of Cryptococcus neoformans were established, compared, and evaluated in murine cryptococcal meningitis. One protocol was designed as a nested PCR to be performed in conventional block thermal cyclers. The other protocol was designed as a quantitative single-round PCR adapted to LightCycler technology. One hundred brain homogenates and dilutions originating from 20 ICR mice treated with different azoles were examined. A fungal burden of 3 × 101 to 2.9 × 104 CFU per mg of brain tissue was determined by quantitative culture. Specific PCR products were amplified by the conventional and the LightCycler methods in 86 and 87 samples, respectively, with products identified by DNA sequencing and real-time fluorescence detection. An analytical sensitivity of 1 CFU of C. neoformans per mg of brain tissue and less than 10 CFU per volume used for extraction was observed for both PCR protocols, while homogenates of 70 organs from mice infected with other fungi were PCR negative. Specificity testing was performed with genomic DNA from 31 hymenomycetous fungal species and from the ustilaginomycetous yeast Malassezia furfur, which are phylogenetically related to C. neoformans. Twenty-four strains, including species of human skin flora like M. furfur and Trichosporon spp., were PCR negative. Amplification was observed with Cryptococcus amylolentus, Filobasidiella depauperata, Cryptococcus laurentii, and five species unrelated to clinical specimens. LightCycler PCR products from F. depauperata and Trichosporon faecale could be clearly discriminated by melting curve analysis. The sensitive and specific nested PCR assay as well as the rapid and quantitative LightCycler PCR assay might be useful for the diagnosis and monitoring of human cryptococcal infections. PMID:11874894

  14. Histopathological study of the effects of low-intensity laser irradiation ({lambda}=650 nm) on dental pulp tissue after cavity preparation; Estudo histopatologico dos efeitos da irradiacao laser em baixa intensidade ({lambda}=650nm) em tecido pulpar apos preparo cavitario

    Energy Technology Data Exchange (ETDEWEB)

    Bertella, Claudio

    2001-07-01

    The purpose of this study was to evaluate (in vivo) the effects of low-intensity Arsenide Gallium Aluminium laser application post-cavity preparation class 1. Six patients with bilateral pre-molars and molars indicated to extraction for orthodontics aim were selected. Four samples of these teeth underwent cavity preparation with deep from medium to high and two samples underwent cavity preparation from high to pulp expositions. The samples were constituted of two teeth of the same patient and received different treatments. One of the teeth underwent laser exposition and the other one was maintained as control, before restorative procedure with polycarboxylate cement. A diode laser ({lambda}=650 nm), output power 30 mW and fluencies of 1,8 J/cm{sup 2} and 2,7 J/cm{sup 2} in pre-molars and molars, respectively, was used for irradiation with repetition rate of 18 Hz in interrupted continuous wave mode. After seven days, the teeth were extracted and processed histologically with HE to verify morphological changes in the pulpy tissue. The four samples, which cavity preparation and restorative material were not in contact with the pulp, did not show histological differences between irradiated and non-irradiated teeth. Both of them presented the same characteristics of normality. The two samples with exposed pulpy tissue showed different results. The irradiated teeth presented no or slight inflammatory signs when compared to the control samples, which showed abscess in the coronary pulp interior and intense inflammatory infiltrated. These results suggest that the laser irradiation can be used as a therapeutic modality in clinical trials, in the conditions employed in this study. (author)

  15. Preservation of nucleic acids and tissue morphology in paraffin-embedded clinical samples: comparison of five molecular fixatives.

    Science.gov (United States)

    Staff, Synnöve; Kujala, Paula; Karhu, Ritva; Rökman, Annika; Ilvesaro, Joanna; Kares, Saara; Isola, Jorma

    2013-09-01

    Formalin fixation preserves tissue morphology at the expense of macromolecule integrity. Freshly frozen samples are the golden standard for DNA and RNA analyses but require laborious deep-freezing and frozen sectioning for morphological studies. Alternative tissue stabilisation methods are therefore needed. We analysed the preservation of nucleic acids, immunohistochemical staining properties and tissue morphology in paraffin-embedded clinical tissue samples fixed with Z7, RCL2, PAXgene, Allprotect and RNAlater. Formalin-fixed and deep-frozen samples were used as controls. Immunohistochemical analyses showed good preservation of antigenicity in all except Allprotect and RNAlater-fixed samples. RNA quality, based on RNA integrity number value by Bioanalyzer, was comparable with freshly frozen samples only in PAXgene-fixed samples. According to quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analyses, RNA from PAXgene samples yielded results similar to freshly frozen samples. No difference between fixatives was seen in DNA analyses (PCR and real-time PCR). In conclusion, PAXgene seems to be superior to other molecular fixatives and formaldehyde.

  16. Quality control in diagnostic molecular pathology in the Netherlands; proficiency testing for patient identification in tissue samples.

    NARCIS (Netherlands)

    Thunnissen, F.B.J.M.; Tilanus, M.G.J.; Ligtenberg, M.J.L.; Nederlof, P.M.; Dinjens, W.N.; Meulemans, E.; Brule, A.J. van den; Noesel, C.J. van; Leeuw, W. de; Schuuring, E.

    2004-01-01

    AIMS: To describe the evolution of proficiency testing for molecular diagnostic pathology with respect to determining unambiguously the patient identity of tissue samples by microsatellite analysis. METHOD: Four rounds of quality control exchanges of samples from different patients were sent with th

  17. Histopathology Image Analysis in Two Long-Term Animal Experiments with Helical Flow Total Artificial Heart.

    Science.gov (United States)

    Wotke, Jiri; Homolka, Pavel; Vasku, Jaromír; Dobsak, Petr; Palanova, Petra; Mrkvicova, Veronika; Konecny, Petr; Soska, Vladimir; Pohanka, Michal; Novakova, Marie; Yurimoto, Terumi; Saito, Itsuro; Inoue, Yusuke; Isoyama, Takashi; Abe, Yusuke

    2016-12-01

    Histopathological analysis can provide important information in long-term experiments with total artificial heart (TAH). Recently, a new type of blood pump, the helical flow total artificial heart (HF-TAH) was developed. This study aimed to investigate the changes in selected vital organs in animal experiments with implanted HF-TAH. Samples from lung, liver, and kidneys from two female goats (No. 1301 and No. 1304) with implanted HF-TAH were analyzed. Tissue samples were fixed in 10% formaldehyde and 4 µm thick transverse sections were stained with hematoxylin-eosin (HE). Additional staining was done for detection of connective tissue (Masson-Goldner stain) and for detection of iron (hemosiderin) deposits (Perls stain). Sections were scanned at 100× and 500× magnification with a light microscope. Experiment no. 1301 survived 100 days (cause of termination was heavy damage of the right pump); experimental goat no.1304 survived 68 days and was sacrificed due to severe right hydrodynamic bearing malfunction. Histopathological analysis of liver samples proved signs of chronic venostasis with limited focal necrotic zones. Dilated tubules, proteinaceous material in tubular lumen, and hemosiderin deposits were detected in kidney samples. Contamination of the organs by embolized micro-particles was suspected at the autopsy after discovery of visible damage (scratches) of the pump impeller surface (made from titanium alloy) in both experiments. Sporadic deposits of foreign micro-particles (presumably titanium) were observed in most of the analyzed parenchymal organs. However, the described deposits were not in direct connection with inflammatory reactions in the analyzed tissues. Histopathological analysis showed the presence of minimal contamination of the lung, kidney, and liver tissue samples by foreign material (titanium very likely). The analysis showed only limited pathological changes, especially in liver and kidneys, which might be attributed to the influence of

  18. THE HISTOPATHOLOGY OF PSORIASIS

    Directory of Open Access Journals (Sweden)

    C. Mignogna

    2011-09-01

    Full Text Available Psoriasis is a common, chronic, relapsing, papulo-squamous dermatitis, with overlying silvery scales. The scalp, sacral region, and extensor surfaces of extremity are commonly involved, even if flexural and intertriginous areas may be affected in the so-called “inverse psoriasis”. Involvement of nails is frequent. Oral lesions (geographic stomatitis and/or glossitis are commonly described. 5-8% of psoriatic patients develop arthritis. Interphalangeal joints are characteristically involved, but large joints are also affected. From a histological point of view, psoriasis is a dynamic dermatosis that changes during the evolution of an individual lesion; we can classify it in an early stage, advanced stage, and later lesions. Lesions are usually diagnostic only in early stages or near the margin of advancing plaques. Munro microabscesses and Kogoj micropustoles are diagnostic clues of psoriasis, but they aren’t always present. All other features can be found in numerous eczematous dermatitis. Key words: Psoriasis, histopathology, immunohistochemistry

  19. Gene expression profiling of human breast tissue samples using SAGE-Seq.

    Science.gov (United States)

    Wu, Zhenhua Jeremy; Meyer, Clifford A; Choudhury, Sibgat; Shipitsin, Michail; Maruyama, Reo; Bessarabova, Marina; Nikolskaya, Tatiana; Sukumar, Saraswati; Schwartzman, Armin; Liu, Jun S; Polyak, Kornelia; Liu, X Shirley

    2010-12-01

    We present a powerful application of ultra high-throughput sequencing, SAGE-Seq, for the accurate quantification of normal and neoplastic mammary epithelial cell transcriptomes. We develop data analysis pipelines that allow the mapping of sense and antisense strands of mitochondrial and RefSeq genes, the normalization between libraries, and the identification of differentially expressed genes. We find that the diversity of cancer transcriptomes is significantly higher than that of normal cells. Our analysis indicates that transcript discovery plateaus at 10 million reads/sample, and suggests a minimum desired sequencing depth around five million reads. Comparison of SAGE-Seq and traditional SAGE on normal and cancerous breast tissues reveals higher sensitivity of SAGE-Seq to detect less-abundant genes, including those encoding for known breast cancer-related transcription factors and G protein-coupled receptors (GPCRs). SAGE-Seq is able to identify genes and pathways abnormally activated in breast cancer that traditional SAGE failed to call. SAGE-Seq is a powerful method for the identification of biomarkers and therapeutic targets in human disease.

  20. A comparison of sample preparation strategies for biological tissues and subsequent trace element analysis using LA-ICP-MS.

    Science.gov (United States)

    Bonta, Maximilian; Török, Szilvia; Hegedus, Balazs; Döme, Balazs; Limbeck, Andreas

    2017-03-01

    Laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) is one of the most commonly applied methods for lateral trace element distribution analysis in medical studies. Many improvements of the technique regarding quantification and achievable lateral resolution have been achieved in the last years. Nevertheless, sample preparation is also of major importance and the optimal sample preparation strategy still has not been defined. While conventional histology knows a number of sample pre-treatment strategies, little is known about the effect of these approaches on the lateral distributions of elements and/or their quantities in tissues. The technique of formalin fixation and paraffin embedding (FFPE) has emerged as the gold standard in tissue preparation. However, the potential use for elemental distribution studies is questionable due to a large number of sample preparation steps. In this work, LA-ICP-MS was used to examine the applicability of the FFPE sample preparation approach for elemental distribution studies. Qualitative elemental distributions as well as quantitative concentrations in cryo-cut tissues as well as FFPE samples were compared. Results showed that some metals (especially Na and K) are severely affected by the FFPE process, whereas others (e.g., Mn, Ni) are less influenced. Based on these results, a general recommendation can be given: FFPE samples are completely unsuitable for the analysis of alkaline metals. When analyzing transition metals, FFPE samples can give comparable results to snap-frozen tissues. Graphical abstract Sample preparation strategies for biological tissues are compared with regard to the elemental distributions and average trace element concentrations.

  1. Normalization of gene expression measurement of tissue samples obtained by transurethral resection of bladder tumors

    Directory of Open Access Journals (Sweden)

    Pop LA

    2016-06-01

    Full Text Available Laura A Pop,1,* Valentina Pileczki,1,2,* Roxana M Cojocneanu-Petric,1 Bogdan Petrut,3,4 Cornelia Braicu,1 Ancuta M Jurj,1 Rares Buiga,5 Patriciu Achimas-Cadariu,6,7 Ioana Berindan-Neagoe1,8 1The Research Center for Functional Genomics, Biomedicine and Translational Medicine, Iuliu Haţieganu University of Medicine and Pharmacy, Cluj-Napoca, Cluj, Romania; 2Department of Analytical Chemistry, Iuliu Haţieganu University of Medicine and Pharmacy, Cluj-Napoca, Cluj, Romania; 3Department of Surgery II – Urology, The Oncology Institute “Prof Dr Ion Chiricuţă”, Cluj-Napoca, Cluj, Romania; 4Department of Urology, Iuliu Haţieganu University of Medicine and Pharmacy, Cluj-Napoca, Cluj, Romania; 5Department of Pathology, The Oncology Institute “Prof. Dr Ion Chiricuţă”, Cluj-Napoca, Cluj, Romania; 6Department of Surgery, The Oncology Institute “Prof Dr Ion Chiricuţă”, Cluj-Napoca, Cluj, Romania; 7Department of Surgical Oncology and Gynecological Oncology, Iuliu Haţieganu University of Medicine and Pharmacy, 8Department of Functional Genomics and Experimental Pathology, The Oncology Institute “Prof Dr Ion Chiricuţă”, Cluj-Napoca, Cluj, Romania *These authors contributed equally to this work Background: Sample processing is a crucial step for all types of genomic studies. A major challenge for researchers is to understand and predict how RNA quality affects the identification of transcriptional differences (by introducing either false-positive or false-negative errors. Nanotechnologies help improve the quality and quantity control for gene expression studies. Patients and methods: The study was performed on 14 tumor and matched normal pairs of tissue from patients with bladder urothelial carcinomas. We assessed the RNA quantity by using the NanoDrop spectrophotometer and the quality by nano-microfluidic capillary electrophoresis technology provided by Agilent 2100 Bioanalyzer. We evaluated the amplification status of three

  2. Grinding and polishing instead of sectioning for the tissue samples with a graft: Implications for light and electron microscopy.

    Science.gov (United States)

    Mukhamadiyarov, Rinat A; Sevostyanova, Victoria V; Shishkova, Daria K; Nokhrin, Andrey V; Sidorova, Olga D; Kutikhin, Anton G

    2016-06-01

    A broad use of the graft replacement requires a detailed investigation of the host-graft interaction, including both histological examination and electron microscopy. A high quality sectioning of the host tissue with a graft seems to be complicated; in addition, it is difficult to examine the same tissue area by both of the mentioned microscopy techniques. To solve these problems, we developed a new technique of epoxy resin embedding with the further grinding, polishing, and staining. Graft-containing tissues prepared by grinding and polishing preserved their structure; however, sectioning frequently required the explantation of the graft and led to tissue disintegration. Moreover, stained samples prepared by grinding and polishing may then be assessed by both light microscopy and backscattered scanning electron microscopy. Therefore, grinding and polishing outperform sectioning when applied to the tissues with a graft.

  3. Discordant assessment of tumor biomarkers by histopathological and molecular assays in the EORTC randomized controlled 10041/BIG 03-04 MINDACT trial breast cancer : Intratumoral heterogeneity and DCIS or normal tissue components are unlikely to be the cause of discordance.

    Science.gov (United States)

    Viale, Giuseppe; Slaets, Leen; de Snoo, Femke A; Bogaerts, Jan; Russo, Leila; van't Veer, Laura; Rutgers, Emiel J T; Piccart-Gebhart, Martine J; Stork-Sloots, Lisette; Dell'Orto, Patrizia; Glas, Annuska M; Cardoso, Fatima

    2016-02-01

    Accurate identification of breast cancer patients most likely to benefit from adjuvant systemic therapies is crucial. Better understanding of differences between methods can lead to an improved ER, PgR, and HER-2 assessment. The purpose of this preplanned translational research is to investigate the correlation of central IHC/FISH assessments with microarray mRNA readouts of ER, PgR, and HER-2 status in the MINDACT trial and to determine if any discordance could be attributed to intratumoral heterogeneity or the DCIS and normal tissue components in the specimens. MINDACT is an international, prospective, randomized, phase III trial investigating the clinical utility of MammaPrint in selecting patients with early breast cancer for adjuvant chemotherapy (n = 6694 patients). Gene-expression data were obtained by TargetPrint; IHC and/or FISH were assessed centrally (n = 5788; 86 %). Macroscopic and microscopic evaluation of centrally submitted FFPE blocks identified 1427 cases for which the very same sample was submitted for gene-expression analysis. TargetPrint ER had a positive agreement of 98 %, and a negative agreement of 95 % with central pathology. Corresponding figures for PgR were 85 and 94 % and for HER-2 72 and 99 %. Agreement of mRNA versus central protein was not different when the same or a different portion of the tumor tissue was analyzed or when DCIS and/or normal tissue was included in the sample subjected to mRNA assays. This is the first large analysis to assess the discordance rate between protein and mRNA analysis of breast cancer markers, and to look into intratumoral heterogeneity, DCIS, or normal tissue components as a potential cause of discordance. The observed difference between mRNA and protein assessment for PgR and HER-2 needs further research; the present analysis does not support intratumoral heterogeneity or the DCIS and normal tissue components being likely causes of the discordance.

  4. ENO1 Protein Levels in the Tumor Tissues and Circulating Plasma Samples of Non-small Cell Lung Cancer Patients

    Directory of Open Access Journals (Sweden)

    Ying ZHANG

    2010-12-01

    Full Text Available Background and objective Proper tumor markers are useful to diagnosis, prognosis and treatment for lung cancer. The aim of this study is to examine the levels of alpha-enolase (ENO1 protein in the tumor tissues and peripheral plasma samples obtained from non-small cell lung cancer (NSCLC patients, and evaluate its potential clinical significance. Methods The ENO1 protein levels in the tumor tissues and corresponding normal tissues from 16 cases of lung squamous cell carcinoma were analyzed by Western blot. The ENO1 protein levels in the plasma samples from 42 healthy individuals, 34 patients with lung benign disease and 84 patients with NSCLC were measured by double antibody sandwich enzyme-linked immunosorbent assay. Results For 87.5% (14/16 of the patients with lung squamous cell carcinoma, the ENO1 protein level in the tumor tissues was higher than that in the corresponding normal lung tissues. The ENO1 protein level in the plasma of NSCLC patients was significantly higher than that in the plasma of healthy individuals (P=0.031 and patients with lung benign disease (P=0.019. Furthermore, the ENO1 protein level was significantly higher in the plasma of patients with lung adenocarcinoma than that of patients with lung squamous cell carcinoma. Conclusion The elevated levels of ENO1 protein in the tumor tissues and the plasma samples from NSCLC patients indicate ENO1 may be a candidate biomarker of lung cancer.

  5. [Inflammatory process, histopathological aspects].

    Science.gov (United States)

    Diébold, J

    1995-01-01

    Inflammation occurs only in conjunctive tissue and is the result of a close cooperation of various cells: blood platelets, endothelial cells, leucocytes, mast cells, fibroblasts. Successive phases can be recognized, the first is characterized by vascular phenomenons defining the acute phase. The second by cellular reactions defining the chronic or granulomatous phase. Various morphological patterns can be recognized in acute or chronic inflammation. In addition, hypersensitivity is responsible of peculiar morphology of the inflammatory response. After tissue necrosis, tissular debris should be eliminated by detersion. Then, a granulation tissue develops representing the first step of the healing, which will not be described here.

  6. [Bacteria isolated from urine and renal tissue samples and their relation to renal histology].

    Science.gov (United States)

    Gökalp, A; Gültekin, E Y; Bakici, M Z; Ozdeşlik, B

    1988-01-01

    The bacteria from the urine and renal biopsy specimens of 40 patients undergoing renal surgery were isolated and their relations with renal histology investigated. The urine cultures were positive in 14 patients, the same organisms being isolated from the renal tissue in 7 cases. In 6 patients with negative urine cultures, bacteria were isolated from renal tissues. Of the 28 cases pathologically diagnosed as chronic pyelonephritis, bacteria were isolated from the renal tissue in 13 cases, the urine cultures being positive in only 11 cases. E. coli was the most commonly encountered bacteria in both the urine and renal tissues.

  7. Histopathologic Comparison of Radiofrequency and Scalpel Blade on Mice Skin Incisional Wound Healing Model

    Directory of Open Access Journals (Sweden)

    Banafshe Gholamhoseini*, Soroush Mohitmafi1, Payam Razeghi Tehrani2 and Sattar Noorialeagha3

    2012-06-01

    Full Text Available Using a safe and accurate alternative to scalpel blades for various surgical operations has been more considering today. There are many studies about the healing process of various tissues after using different methods like radiofrequency, electro surgery and scalpel blade for the surgical incisions, but choosing the right instrument is very important in cosmetic surgeries.This study was undertaken to get more histopathologic details about the healing process after using either Radiofrequency (RF or Scalpel blade as a skin incision method. So, an experimental study was designed in that 12 mice were randomly divided in three groups with the same situation and two different incisions were cut in each mouse on ventral skin by scalpel blade and radio surgical equipment. The skin specimens from scalpel blade incision as group one sample and radiosurgical incision as group two sample were harvested on the same operation day and a weekly manner to day 21. All samples were stained with Hematoxillin - Eosin and observed under light microscopy. Results showed less tissue contraction at the cutting edge and better collagen alignments in RF incision in comparison to scalpel blade incisions in which having fibrosis and more tissue contraction. These histopathologic findings might be the reason for more elasticity and less scar tissue remaining after skin Radiofrequency surgery.

  8. Vibrational spectroscopy studies of formalin-fixed cervix tissues.

    Science.gov (United States)

    Krishna, C M; Sockalingum, G D; Vadhiraja, B M; Maheedhar, K; Rao, A C K; Rao, L; Venteo, L; Pluot, M; Fernandes, D J; Vidyasagar, M S; Kartha, V B; Manfait, M

    2007-02-15

    Optical histopathology is fast emerging as a potential tool in cancer diagnosis. Fresh tissues in saline are ideal samples for optical histopathology. However, evaluation of suitability of ex vivo handled tissues is necessitated because of severe constraints in sample procurement, handling, and other associated problems with fresh tissues. Among these methods, formalin-fixed samples are shown to be suitable for optical histopathology. However, it is necessary to further evaluate this method from the point of view discriminating tissues with minute biochemical variations. A pilot Raman and Fourier transform infrared (FTIR) microspectroscopic studies of formalin-fixed tissues normal, malignant, and after-2-fractions of radiotherapy from the same malignant cervix subjects were carried out, with an aim to explore the feasibility of discriminating these tissues, especially the tissues after-2-fractions of radiotherapy from other two groups. Raman and FTIR spectra exhibit large differences for normal and malignant tissues and subtle differences are seen between malignant and after-2-fractions of radiotherapy tissues. Spectral data were analyzed by principal component analysis (PCA) and it provided good discrimination of normal and malignant tissues. PCA of data of three tissues, normal, malignant, and 2-fractions after radiotherapy, gave two clusters corresponding to normal and malignant + after-2-fractions of radiotherapy tissues. A second step of PCA was required to achieve discrimination between malignant and after-2-fractions of radiotherapy tissues. Hence, this study not only further supports the use of formalin-fixed tissues in optical histopathology, especially from Raman spectroscopy point of view, it also indicates feasibility of discriminating tissues with minute biochemical differences such as malignant and after-2-fractions of radiotherapy.

  9. Epigenome-wide profiling of DNA methylation in paired samples of adipose tissue and blood.

    Science.gov (United States)

    Huang, Yen-Tsung; Chu, Su; Loucks, Eric B; Lin, Chien-Ling; Eaton, Charles B; Buka, Stephen L; Kelsey, Karl T

    2016-03-03

    Many epigenetic association studies have attempted to identify DNA methylation markers in blood that are able to mirror those in target tissues. Although some have suggested potential utility of surrogate epigenetic markers in blood, few studies have collected data to directly compare DNA methylation across tissues from the same individuals. Here, epigenomic data were collected from adipose tissue and blood in 143 subjects using Illumina HumanMethylation450 BeadChip array. The top axis of epigenome-wide variation differentiates adipose tissue from blood, which is confirmed internally using cross-validation and externally with independent data from the two tissues. We identified 1,285 discordant genes and 1,961 concordant genes between blood and adipose tissue. RNA expression data of the two classes of genes show consistent patterns with those observed in DNA methylation. The discordant genes are enriched in biological functions related to immune response, leukocyte activation or differentiation, and blood coagulation. We distinguish the CpG-specific correlation from the within-subject correlation and emphasize that the magnitude of within-subject correlation does not guarantee the utility of surrogate epigenetic markers. The study reinforces the critical role of DNA methylation in regulating gene expression and cellular phenotypes across tissues, and highlights the caveats of using methylation markers in blood to mirror the corresponding profile in the target tissue.

  10. A pilot study of sampling subcutaneous adipose tissue to examine biomarkers of cancer risk

    OpenAIRE

    Campbell, Kristin L.; Makar, Karen W.; Kratz, Mario; Foster-Schubert, Karen E.; McTiernan, Anne; Ulrich, Cornelia M.

    2009-01-01

    Examination of adipose tissue biology may provide important insight into mechanistic links for the observed association between higher body fat and risk of several types of cancer, in particular colorectal and breast cancer. We tested two different methods of obtaining adipose tissue from healthy individuals.

  11. Evaluation of lethal and non-lethal sampling methods for the detection of white sturgeon iridovirus infection in white sturgeon, Acipenser transmontanus (Richardson).

    Science.gov (United States)

    Drennan, J D; Lapatra, S E; Samson, C A; Ireland, S; Eversman, K F; Cain, K D

    2007-06-01

    Pectoral fin tissue of white sturgeon was investigated as a potential non-lethal sample source for the detection of white sturgeon iridovirus (WSIV) infection. Histopathology and polymerase chain reaction (PCR) results using fin tissue were compared with the standard lethal histopathology sampling method that utilizes head tissue. Tissues for each of the three sampling methods were collected weekly for 8 weeks from individual sturgeon undergoing an experimental cohabitation challenge with fish infected with the Abernathy isolate of WSIV. Non-lethal fin histopathological evaluation did not reveal infection during the first 3 weeks of sampling, while non-lethal PCR and the lethal method were variable. However, all three sampling methods were equally capable of identifying infection from 4 to 8 weeks post-exposure. Of the survivors tested, all were negative by PCR and the lethal method, and only one fish was identified as being positive by non-lethal fin histopathology. In another experiment, all three sampling methods were applied to asymptomatic WSIV carriers in a case study conducted at the Kootenai Tribal Sturgeon Conservation Hatchery. Results showed that both lethal and non-lethal fin histopathology were equally effective in detecting infection, but PCR was unable to identify this strain of WSIV. Depending on the virus isolate, these results suggest that non-lethal sampling of fin tissue (histopathology or PCR) is comparable with the lethal sampling method at identifying WSIV infection once infection is established, and under certain circumstances may provide an alternative to lethal sampling.

  12. Profiling of adrenocorticotropic hormone and arginine vasopressin in human pituitary gland and tumor thin tissue sections using droplet-based liquid-microjunction surface-sampling-HPLC-ESI-MS-MS.

    Science.gov (United States)

    Kertesz, Vilmos; Calligaris, David; Feldman, Daniel R; Changelian, Armen; Laws, Edward R; Santagata, Sandro; Agar, Nathalie Y R; Van Berkel, Gary J

    2015-08-01

    Described here are the results from the profiling of the proteins arginine vasopressin (AVP) and adrenocorticotropic hormone (ACTH) from normal human pituitary gland and pituitary adenoma tissue sections, using a fully automated droplet-based liquid-microjunction surface-sampling-HPLC-ESI-MS-MS system for spatially resolved sampling, HPLC separation, and mass spectrometric detection. Excellent correlation was found between the protein distribution data obtained with this method and data obtained with matrix-assisted laser desorption/ionization (MALDI) chemical imaging analyses of serial sections of the same tissue. The protein distributions correlated with the visible anatomic pattern of the pituitary gland. AVP was most abundant in the posterior pituitary gland region (neurohypophysis), and ATCH was dominant in the anterior pituitary gland region (adenohypophysis). The relative amounts of AVP and ACTH sampled from a series of ACTH-secreting and non-secreting pituitary adenomas correlated with histopathological evaluation. ACTH was readily detected at significantly higher levels in regions of ACTH-secreting adenomas and in normal anterior adenohypophysis compared with non-secreting adenoma and neurohypophysis. AVP was mostly detected in normal neurohypophysis, as expected. This work reveals that a fully automated droplet-based liquid-microjunction surface-sampling system coupled to HPLC-ESI-MS-MS can be readily used for spatially resolved sampling, separation, detection, and semi-quantitation of physiologically-relevant peptide and protein hormones, including AVP and ACTH, directly from human tissue. In addition, the relative simplicity, rapidity, and specificity of this method support the potential of this basic technology, with further advancement, for assisting surgical decision-making. Graphical Abstract Mass spectrometry based profiling of hormones in human pituitary gland and tumor thin tissue sections.

  13. Genomic DNA isolation of Acrocomia aculeata (Arecaceae) from leaf and stipe tissue samples for PCR analysis.

    Science.gov (United States)

    Lanes, E C M; Nick, C; Kuki, K N; Freitas, R D; Motoike, S Y

    2013-09-23

    Macaw palm, Acrocomia aculeata is an oleaginous species of the Arecaceae family; it has been identified as one of the most promising plants for sustainable production of renewable energy, especially biodiesel. We developed an efficient protocol of genomic DNA extraction for A. aculeata using leaf and stipe tissues, based on the cationic hexadecyltrimethylammonium bromide method, and we evaluated the quantity, purity, and integrity of the resultant DNA. We also determined whether these procedures interfere with PCR amplification using SSR molecular markers. The lowest concentration of DNA was obtained from stipe tissues (135 ng/μL), while fresh leaf tissues provided the highest concentration of DNA (650 ng/μL). Good quality DNA was obtained from fresh leaf, lyophilized leaf, and stipe tissues (relative purity, 1.79-1.89 nm). Differences in quantity and quality of DNA extracted from different tissues did not interfere with general patterns of PCR amplification based on SSR markers.

  14. Comparison of ESR1 Mutations in Tumor Tissue and Matched Plasma Samples from Metastatic Breast Cancer Patients

    Directory of Open Access Journals (Sweden)

    Takashi Takeshita

    2017-10-01

    Full Text Available BACKGROUND: ESR1 mutation in circulating cell-free DNA (cfDNA is emerging as a noninvasive biomarker of acquired resistance to endocrine therapy, but there is a paucity of data comparing the status of ESR1 gene in cfDNA with that in its corresponding tumor tissue. The objective of this study is to validate the degree of concordance of ESR1 mutations between plasma and tumor tissue. METHODS: ESR1 ligand-binding domain mutations Y537S, Y537N, Y537C, and D538G were analyzed using droplet digital PCR in 35 patients with metastatic breast cancer (MBC (35 tumor tissue samples and 67 plasma samples. RESULTS: Of the 35 paired samples, 26 (74.3% were concordant: one patient had detectable ESR1 mutations both plasma (ESR1 Y537S/Y537N and tumor tissue (ESR1 Y537S/Y537C, and 25 had WT ESR1 alleles in both. Nine (25.7% had discordance between the plasma and tissue results: five had mutations detected only in their tumor tissue (two Y537S, one Y537C, one D538G, and one Y537S/Y537N/D538G, and four had mutations detected only in their plasma (one Y537S, one Y537N, and two Y537S/Y537N/D538G. Furthermore, longitudinal plasma samples from 19 patients were used to assess changes in the presence of ESR1 mutations during treatment. Eleven patients had cfDNA ESR1 mutations over the course of treatment. A total of eight of 11 patients with MBC with cfDNA ESR1 mutations (72.7% had the polyclonal mutations. CONCLUSION: We have shown the independent distribution of ESR1 mutations between plasma and tumor tissue in 35 patients with MBC.

  15. [Histopathological meniscus diagnostic].

    Science.gov (United States)

    Fisseler-Eckhoff, A; Müller, K-M

    2009-06-01

    Menisci fulfill many functions within the complex biomechanics of the knee joint. In the case of meniscus lesions, sparing arthroscopic resections and operative refixation are the treatments of choice. With regard to diagnostics, this means that in general terms, the histopathologic diagnostics are carried out on detached meniscus fragments of between 5 mm and 2 cm in size. An experienced pathologist's knowledge of physiologically possible cellular and fibrous histological meniscus damage, as opposed to nonphysiological change regarded as normal with respect to age, is essential during a diagnostic meniscus evaluation. The clinician expects clear statements from the pathologist regarding the severity of previous or secondary degenerative meniscus damage, the age and type of traumatic tears, and appraisal of the relationship between trauma and meniscus damage from an insurance point of view. Close cooperation between the clinician and the pathologist allows for fast and unambiguous correlation of anamnesis, the clinical picture, and morphological reporting so that cases involving insurance problems - which are numerous, often long-term, and often unsatisfactory - can be clarified quickly.

  16. Suitability of faeces and tissue samples as a basis for non-invasive sampling for African swine fever in wild boar.

    Science.gov (United States)

    de Carvalho Ferreira, H C; Weesendorp, E; Quak, S; Stegeman, J A; Loeffen, W L A

    2014-08-27

    A challenging aspect of ASFV control in wild boar populations is the design and implementation of effective surveillance and monitoring programmes, both for early warning, and to determine the ongoing epidemiological situation in an infected population. Testing blood samples requires invasive sampling strategies like hunting or capture of wild boar. Besides being biased towards healthy animals, such strategies are also linked to further spread of the virus. Non-invasive sampling strategies would increase the reliability of surveillance of ASFV in wild boar populations, without the negative side effects. This study evaluates the potential of faeces and tissue samples as a basis for non-invasive sampling strategies for ASFV in wild boar. In the acute phase (0-21 days after infection), in comparison with virus detection in blood, virus can be detected in faeces 50-80% of the time. This percentage decreases to below 10% for the subacute/chronic phase. ASFV DNA is quite stable in faeces. Half-lives range from more than 2 years at temperature up to 12°C, to roughly 15 days at temperatures of 30°C. In tissue samples, stored at 20°C, half-lives mostly range from 1.7 to 7.4 days. The sample of preference is the spleen, where the highest titres and highest half-life of ASFV DNA are observed. The level and duration of excretion of ASFV in the faeces, combined with the stability of the DNA, suggest that sampling of faeces could be the basis for a non-invasive sampling strategy to monitor ASFV in wild boar.

  17. Development and application of specific cytokine assays in tissue samples from a bottlenose dolphin with hyperinsulinemia

    Science.gov (United States)

    Chronic inflammation has been associated with insulin resistance and type 2 diabetes in humans. Postmortem hepatic and splenic tissue from a 46-year old geriatric male bottlenose dolphin (Tursiops truncatus) with insulin resistance (chronic hyperinsulinemia with hyperglycemia) , chronic = inflamma...

  18. Histopathological Study of Trabeculum after Excimer Laser Trabeculectomy ab Interno

    Institute of Scientific and Technical Information of China (English)

    Shengsong Huang; Minbin Yu; Guangqiang Feng; Ping Zhang; Changyu Qiu

    2001-01-01

    Purpose: To study the clinical manifestations and histopathologic changes of trabeculum after excimer laser trabeculectomy ab interno (ELT), and to investigate the mechanisms of ELT in reducing intraocular pressure. Methods : ELT was performed on ten rabbit eves and postoperative responses were documented. Comeoscleral tissue samples were harvested consecutively each week until the 5th postoperative week and these samples were examined under light and electrical microscopy. Results: Mild stimulation signs were present postoperatix'ely in nine of ten eyes, but no serious complications were experienced. Obvious inflammation was observed in one rabbit eye as a result of iris damage during the surgical manipulations. Local kactures on the trabecular meshwork and openings into Schlemm' s canal were detected in all tissue samples under light microscope. Mitochondria were found to be turgescent and dilated like vacuoles and endoplasmic reticula were found to be dilated under electrical microscope in the early postoperative period. Later, all trabecular cells returned normal and no fihroblast cells were ever detected. Conclusions: Permanent openings through trabecular meshwork into the inner wall of Schlemm's canal can be created with ELT. The outflow resistance of aqueous humor can be reduced with these openings and intraocular pressure can be controlled thereafter. Eye Science 2001; 17:11 ~ 15.

  19. Single-pixel hyperspectral imaging for real-time cancer detection: detecting damage in ex vivo porcine tissue samples

    Science.gov (United States)

    Peller, Joseph; Farahi, Faramarz; Trammell, Susan R.

    2016-03-01

    We are developing a single-pixel hyperspectral imaging system based on compressive sensing that acquires spatial and spectral information simultaneously. Our spectral imaging system uses autofluorescencent emission from collagen (400 nm) and NAD(P)H (475 nm), as well as, differences in the optical reflectance spectra as diagnostics for differentiating between healthy and diseased tissue. In this study, we demonstrate the ability of our imaging system to discriminate between healthy and damaged porcine epidermal tissue. Healthy porcine epidermal tissue samples (n=11) were imaged ex vivo using our hyperspectral system. The amount of NAD(P)H emission and the reflectance properties were approximately constant across the surface of healthy tissue samples. The tissue samples were then thermally damaged using an 1850 nm thulium fiber laser and re-imaged after laser irradiation. The damaged regions were clearly visible in the hyperspectral images as the thermal damage altered the fluorescent emission of NAD(P)H and changed the scattering properties of the tissue. The extent of the damaged regions was determined based on the hyperspectral images and these estimates were compared to damage extents measured in white light images acquired with a traditional camera. The extent of damage determined via hyperspectral imaging was in good agreement with estimates based on white light imaging indicating that our system is capable of differentiating between healthy and damaged tissue. Possible applications of our single pixel hyperspectral imaging system range from real-time determination of tumor margins during surgery to the use of this technique in the pathology lab to aid with cancer diagnosis and staging.

  20. Sugar and acid interconversion in tomato fruits based on biopsy sampling of locule gel and pericarp tissue

    NARCIS (Netherlands)

    Schouten, R.E.; Woltering, E.J.; Tijskens, L.M.M.

    2016-01-01

    This study deals with quantifying sugar and acids levels important for the perceived taste of tomatoes (Solanum lycopersicum). Sugar and acids levels were measured repeatedly on the same tomato using tissue samples obtained with a biopsy needle in combination with HPLC protocols. Biopsies of

  1. Comparative evaluation of specific methods for labeling of Encephalitozoon cuniculi in paraffin wax-embedded tissue samples.

    Science.gov (United States)

    Habenbacher, Bettina; Klang, Andrea; Fragner, Karin; Dinhopl, Nora; Künzel, Frank; Weissenböck, Herbert

    2012-03-01

    Detection of the microsporidian Encephalitozoon cuniculi in tissue samples is considered difficult. The aim of the current study was to determine whether immunohistochemistry (IHC) and in situ hybridization (ISH) represent reliable methods for the detection of E. cuniculi in postmortem tissue samples of rabbits. Paraffin-embedded tissue sections of brain and kidneys of 48 naturally infected pet rabbits, 10 negative controls, and the eyes of 3 further rabbits were used for all investigations. By IHC in 19 animals (37.3%), spores could be clearly detected and were all equally stained. By ISH using a digoxigenin-labeled oligonucleotide probe, only 6 animals (11.8%) proved undoubtedly positive. In these cases, many parasite-like objects revealed strong typical purple-black positive signals. However, several of the examined samples showed only partial staining of the pathogen or unclear results. Thus, in order to find an explanation for these inconsistent ISH results and to take a more detailed look at the different developmental stages of the organism, electron microscopy was applied. Empty spores, which had already discharged their polar filaments, prevailed in total number. Taken together, both techniques are rather insensitive, but under the condition that sufficient numbers of microsporidia are present, IHC can be recommended for specific identification of E. cuniculi in tissue samples. In contrast, ISH failed to detect some developmental stages of the organism, and, as such, ISH is therefore considered an inappropriate diagnostic method.

  2. A comparison of liver sampling techniques in dogs.

    Science.gov (United States)

    Kemp, S D; Zimmerman, K L; Panciera, D L; Monroe, W E; Leib, M S; Lanz, O I

    2015-01-01

    The liver sampling technique in dogs that consistently provides samples adequate for accurate histopathologic interpretation is not known. To compare histopathologic results of liver samples obtained by punch, cup, and 14 gauge needle to large wedge samples collected at necropsy. Seventy dogs undergoing necropsy. Prospective study. Liver specimens were obtained from the left lateral liver lobe with an 8 mm punch, a 5 mm cup, and a 14 gauge needle. After sample acquisition, two larger tissue samples were collected near the center of the left lateral lobe to be used as a histologic standard for comparison. Histopathologic features and numbers of portal triads in each sample were recorded. The mean number of portal triads obtained by each sampling method were 2.9 in needle samples, 3.4 in cup samples, 12 in punch samples, and 30.7 in the necropsy samples. The diagnoses in 66% of needle samples, 60% of cup samples, and 69% of punch samples were in agreement with the necropsy samples, and these proportions were not significantly different from each other. The corresponding kappa coefficients were 0.59 for needle biopsies, 0.52 for cup biopsies, and 0.62 for punch biopsies. The histopathologic interpretation of a liver sample in the dog is unlikely to vary if the liver biopsy specimen contains at least 3-12 portal triads. However, in comparison large necropsy samples, the accuracy of all tested methods was relatively low. Copyright © 2014 by the American College of Veterinary Internal Medicine.

  3. Comparison PCR Method and Rapid Urease Test to Detect Helicobacter Pylori in the Gastric Biopsy Tissue Samples

    Directory of Open Access Journals (Sweden)

    Parastoo Chamanrokh

    2013-09-01

    Full Text Available Background & Objective: Helicobacter pylori has been recognized as a major risk factor in gastric and duodenum ulcers and gastric cancer. Some laboratory tests, such as culture, are not entirely satisfying. The aim of this study is to compare RUT with PCR in identifying H. pylori in the gastric biopsy tissue samples.Materials & Methods: First, the standard H.pylori sample (N:oC30 was provided. primers or the glmM (ureC gene were used In PCR method The PCR test was optimized by sensitivity and specificity methods. Then 100 clinical samples composed of stomach tissue biopsy were prepared. The rapid urease test was conducted on all obtained samples to identify H.pylori. DNA was extracted from samples using DNG plus technique. Then, samples were studied using PCR method.  Results: The 294 bp product was amplified in the optimized PCR test. The PCR test sensitivity was obtained at 10 CFU. Any unwanted products were not seen in the specificity test with the exception of H. pylori DNA samples. Of 100 stomach biopsy samples, 64% were reported as positive by RUT and 76% by the PCR test.Conclusion: Given that the PCR test has higher sensitivity and specificity to detect H.pylori comparing rapid ureaas test, therefore this method could be used to detect H.pylori.  

  4. Probing focal cortical dysplasia in formalin fixed samples using tissue optical spectroscopy

    Science.gov (United States)

    Anand, Suresh; Cicchi, Riccardo; Giordano, Flavio; Buccoliero, Anna Maria; Conti, Valerio; Guerrini, Renzo; Pavone, Francesco Saverio

    2016-03-01

    Focal cortical dysplasia (FCD) is one of most common causes of intractable epilepsy in pediatric population and these are often insensitive to anti-epileptic drugs. FCD is characterized by a disarray in localized regions of the cerebral cortex and abnormal neurons which results them to misfire with incorrect signals. Resective neurosurgery to remove or disconnect the affected parts from the rest of the brain seems to be a viable option to treat FCD. Before neurosurgery the subject could undergo imaging studies including magnetic resonance imaging (MRI) or computed tomography (CT) scans. On the downside FCD could be elusive in MRI images and may be practically invisible in CT scans. Furthermore, unnecessary removal of normal tissues is to be taken into consideration as this could lead to neurological defects. In this context, optical spectroscopy have been widely investigated as an alternative technique for the detection of abnormal tissues in different organ sites. Disease progression is accompanied by a number of architectural, biochemical and morphological changes. These variations are reflected in the spectral intensity and line shape. Here, in this proof of concept study we propose to investigate the application of tissue optical spectroscopy based on fluorescence excitation at two wavelength 378 and 445 nm coupled along with Raman spectroscopy for the detection of FCD on formalin fixed tissue specimens from pediatric subjects. For fluorescence at both the excitation wavelengths FCD showed a decreased intensity at longer wavelength when compared to normal tissues. Also, differences exist in the Raman spectral profiles of normal and FCD.

  5. Comparing citrated native, kaolin-activated, and tissue factor-activated samples and determining intraindividual variability for feline thromboelastography.

    Science.gov (United States)

    Banerjee, Amrita; Blois, Shauna L; Wood, R Darren

    2011-11-01

    Thromboelastography (TEG) is a point-of-care whole blood test of hemostasis. While TEG is becoming more widely used in veterinary medicine, few studies describe the use of TEG in cats. The objectives of the current study were to: 1) document the range of TEG variables produced in healthy cats using 3 sample types (citrated native, kaolin-activated, and tissue factor-activated), and 2) determine if there was a significant difference between 2 separate samples obtained from individual healthy cats on the same day. Jugular venipuncture was performed in 20 cats, and citrated blood collected for TEG. TEG analysis was performed on citrated native, kaolin-activated, and tissue factor-activated blood for each sample. Two hours later, the procedure was repeated from the opposite jugular vein, yielding a total of 120 analyses. Reaction time (R), alpha angle (α), kappa value (κ), and maximum amplitude (MA) were recorded from each tracing. No significant differences were found between TEG tracings from the first and second venipuncture samples. Significant differences were found between sample types for R, α, κ, and MA. Means for citrated native/kaolin-activated/tissue factor-activated methods were R = 4.1/3.7/0.6 min; κ = 2.5/1.8/2.2 min; α = 59.9/65.1/70.4 degrees; MA = 47.4/49.9/44.7 mm. A limitation of this study was the small number of cats used. Thromboelastography analysis may be a suitable method of evaluating hemostasis in cats.

  6. Comparing the proteome of snap frozen, RNAlater preserved, and formalin-fixed paraffin-embedded human tissue samples

    DEFF Research Database (Denmark)

    Bennike, Tue Bjerg; Kastaniegaard, Kenneth; Padurariu, Simona

    2016-01-01

    Large biobanks exist worldwide containing formalin-fixed, paraffin-embedded samples and samples stored in RNAlater. However, the impact of tissue preservation on the result of a quantative proteome analysis remains poorly described. Human colon mucosal biopsies were extracted from the sigmoideum...... and either immediately frozen, stabilized in RNAlater, or stabilized by formalin-fixation. In one set of biopsies, formalin stabilization was delayed for 30 min. The protein content of the samples was characterized by high throughput quantitative proteomics. We were able to identify a similar high number...

  7. Quantification of Toxoplasma gondii in tissue samples of experimentally infected goats by magnetic capture and real-time PCR.

    Science.gov (United States)

    Juránková, Jana; Opsteegh, Marieke; Neumayerová, Helena; Kovařčík, Kamil; Frencová, Anita; Baláž, Vojtěch; Volf, Jiří; Koudela, Břetislav

    2013-03-31

    Undercooked meat containing tissue cysts is one of the most common sources of Toxoplasma gondii infection in humans. Goats are very susceptible to clinical toxoplasmosis, and especially kids are common food animals, thereby representing a risk for human infection. A sequence-specific magnetic capture method was used for isolation of T. gondii DNA from tissue samples from experimentally infected goat-kids and real-time PCR for the 529 bp repeat element allowed quantification of T. gondii DNA. The contamination level in different types of tissue and in two groups of goats euthanized 30 and 90 dpi was compared. The highest concentration of T. gondii DNA in both groups of goats was found in lung tissue, but only the higher parasite count in lung tissue compared to other organs in group A (euthanized 30 dpi) was statistically significant. T. gondii concentrations were higher in liver and dorsal muscle samples from goats euthanized 90 dpi than in goats euthanized at 30 dpi, while the T. gondii concentration in hearts decreased. This study describes for the first time distribution of T. gondii parasites in post-weaned goat kids. New information about T. gondii predilection sites in goats and about the progression of infection between 30 and 90 dpi was achieved. Copyright © 2012 Elsevier B.V. All rights reserved.

  8. Histopathologic criteria to confirm white-nose syndrome in bats

    Science.gov (United States)

    Meteyer, C.U.; Buckles, E.L.; Blehert, D.S.; Hicks, A.C.; Green, D.E.; Shearn-Bochsler, V.; Thomas, N.J.; Gargas, A.; Behr, M.J.

    2009-01-01

    White-nose syndrome (WNS) is a cutaneous fungal disease of hibernating bats associated with a novel Geomyces sp. fungus. Currently, confirmation of WNS requires histopathologic examination. Invasion of living tissue distinguishes this fungal infection from those caused by conventional transmissible dermatophytes. Although fungal hyphae penetrate the connective tissue of glabrous skin and muzzle, there is typically no cellular inflammatory response in hibernating bats. Preferred tissue samples to diagnose this fungal infection are rostral muzzle with nose and wing membrane fixed in 10% neutral buffered formalin. To optimize detection, the muzzle is trimmed longitudinally, the wing membrane is rolled, and multiple cross-sections are embedded to increase the surface area examined. Periodic acid-Schiff stain is essential to discriminate the nonpigmented fungal hyphae and conidia. Fungal hyphae form cup-like epidermal erosions and ulcers in the wing membrane and pinna with involvement of underlying connective tissue. In addition, fungal hyphae are present in hair follicles and in sebaceous and apocrine glands of the muzzle with invasion of tissue surrounding adnexa. Fungal hyphae in tissues are branching and septate, but the diameter and shape of the hyphae may vary from parallel walls measuring 2 ??m in diameter to irregular walls measuring 3-5 ??m in diameter. When present on short aerial hyphae, curved conidia are approximately 2.5 ??m wide and 7.5 ??m in curved length. Conidia have a more deeply basophilic center, and one or both ends are usually blunt. Although WNS is a disease of hibernating bats, severe wing damage due to fungal hyphae may be seen in bats that have recently emerged from hibernation. These recently emerged bats also have a robust suppurative inflammatory response.

  9. Enhanced visualization of histological samples with an adjustable RGB contrast system with application for tissue used in photodynamic therapy.

    Science.gov (United States)

    Barrionuevo, Wilma Regina; Filho, Edson Cesar Marques; Bagnato, Vanderlei Salvador

    2008-06-01

    The analysis of histological sections has long been a valuable tool in the pathological studies. The interpretation of tissue conditions, however, relies directly on visual evaluation of tissue slides, which may be difficult to interpret because of poor contrast or poor color differentiation. The Chromatic Contrast Visualization System (CCV) combines an optical microscope with electronically controlled light-emitting diodes (LEDs) in order to generate adjustable intensities of RGB channels for sample illumination. While most image enhancement techniques rely on software post-processing of an image acquired under standard illumination conditions, CCV produces real-time variations in the color composition of the light source itself. The possibility of covering the entire RGB chromatic range, combined with the optical properties of the different tissues, allows for a substantial enhancement in image details. Traditional image acquisition methods do not exploit these visual enhancements which results in poorer visual distinction among tissue structures. Photodynamic therapy (PDT) procedures are of increasing interest in the treatment of several forms of cancer. This study uses histological slides of rat liver samples that were induced to necrosis after being exposed to PDT. Results show that visualization of tissue structures could be improved by changing colors and intensities of the microscope light source. PDT-necrosed tissue samples are better differentiated when illuminated with different color wavelengths, leading to an improved differentiation of cells in the necrosis area. Due to the potential benefits it can bring to interpretation and diagnosis, further research in this field could make CCV an attractive technique for medical applications.

  10. Optical parameter measurement of highly diffusive tissue body phantoms with specifically designed sample holder for photo diagnostic and PDT applications

    Science.gov (United States)

    Rehman, A.; Rehman, K.; Anwar, S.; Firdous, S.; Nawaz, M.

    2015-12-01

    Knowledge of optical properties (absorption coefficients, scattering Coefficients, and anisotropy) is necessary for understanding light tissue interactions. Optical parameters define the behavior of light in the tissues. Intralipid and Indian ink are well-established tissue body phantoms. Quantitative characterization of biological tissues in terms of optical properties is achieved with integrating sphere. However, samples having significantly higher scattering and absorption coefficients such as malignant tissues potentially reduce the signal to noise ratio (SNR) and accuracy of integrating sphere. We have measured the diffuse reflection and transmission of these phantoms by placing them in integrating sphere at 632.8 nm and then applied IAD method to determine the optical properties tissue phantoms composed of Indian ink (1.0%) and Intralipid (20%). We have fabricated a special sample holder with thin microscopic cover slips which can be used to measure signal from highly concentrated intralipid and Indian ink solutions. Experiments conducted with various phantoms reveal significant improvement of SNR for a wide range of optical properties. This approach opens up a field for potential applications in measurement of optical properties of highly diffusive biological tissues. For 20% intralipid μa =0.112+/-0.046 cm-1 and μs =392.299+/-10.090 cm-1 at 632.8 nm and for 1.0% Indian ink μa =9.808+/-0.490 cm-1 and μs =1.258+/-0.063 cm-1 at same wavelength. System shows good repeatability and reproducibility within 4.9% error. Work may have important biomedical applications in photo-diagnosis and Photodynamic therapy.

  11. Comprovação anatomopatológica da presença de tecido vascular e nervoso evidenciado à ecografia como eco tangencial ao descolamento de coróide: relato de dois casos Histopathologic confirmation of vascular and nervous tissue identified on ultrasound as an tangential image to choroidal detachment: report of two cases

    Directory of Open Access Journals (Sweden)

    Eduardo F. Marback

    2009-12-01

    Full Text Available São apresentados dois casos de descolamento de coróide em olhos com endoftalmite submetidos a enucleação. O estudo anatomopatológico permitiu identificar a presença de tecido vascular e nervoso interpondo-se entre a esclera e coróide deslocada, como havia sido evidenciado pela ecografia.Two cases of endophthalmitis with choroidal detachment submitted to enucleation are presented. The histopathologic study allowed the identification of vascular and nervous tissue between the choroid and sclera, as it was seen on ultrasound examination.

  12. Measurement of intravenously administered γ-Fe2O3 particle amount in mice tissues using vibrating sample magnetometer.

    Science.gov (United States)

    Kishimoto, Mikio; Miyamoto, Ryoichi; Oda, Tatsuya; Ohara, Yusuke; Yanagihara, Hideto; Ohkohchi, Nobuhiro; Kita, Eiji

    2014-12-01

    Dispersions of platelet γ-Fe2O3 particles 30-50nm in size were intravenously administered to mice and the amount of particles accumulated in each tissue was obtained by magnetization measurement using a vibrating sample magnetometer. Background noise was greatly reduced by measuring dried tissues under a magnetic field of 500 Oe so that the effect of diamagnetism was slight. Remarkable particle accumulation was observed in the liver and spleen. Considerable particle accumulation was observed in the lung when a large quantity of γ-Fe2 O3 particles was administered. There was no significant particle accumulation in the kidney and heart.

  13. Experimental evaluation and computational modeling of tissue damage from low-flow push-pull perfusion sampling in vivo.

    Science.gov (United States)

    Cepeda, David E; Hains, Leah; Li, David; Bull, Joseph; Lentz, Stephen I; Kennedy, Robert T

    2015-03-15

    Neurochemical monitoring via sampling probes is valuable for deciphering neurotransmission in vivo. Microdialysis is commonly used; however, the spatial resolution is poor. Recently push-pull perfusion at low flow rates (50nL/min) has been proposed as a method for in vivo sampling from the central nervous system. Tissue damage from such probes has not been investigated in detail. In this work, we evaluated acute tissue response to low-flow push-pull perfusion by infusing the nuclear stains Sytox Orange and Hoechst 33342 through probes implanted in the striatum for 200min, to label damaged and total cells, respectively, in situ. Using the damaged/total labeled cell ratio as a measure of tissue damage, we found that 33±8% were damaged within the dye region around a microdialysis probe. We found that low-flow push-pull perfusion probes damaged 24±4% of cells in the sampling area. Flow had no effect on the number of damaged cells for low-flow push-pull perfusion. Modeling revealed that shear stress and pressure gradients generated by the flow were lower than thresholds expected to cause damage. Comparison with existing methods.Push-pull perfusion caused less tissue damage but yielded 1500-fold better spatial resolution. Push-pull perfusion at low flow rates is a viable method for sampling from the brain with potential for high temporal and spatial resolution. Tissue damage is mostly caused by probe insertion. Smaller probes may yield even lower damage. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. A novel approach in personal identification from tissue samples undergone different processes through STR typing.

    Science.gov (United States)

    Staiti, N; Di Martino, D; Saravo, L

    2004-12-01

    Short tandem repeats (STRs) or microsatellites have been recognized worldwide as a powerful tool for human identification. They have become widely used in human identification especially in criminal cases and mass disasters. Police departments are often interested in cases where tissues are already decomposed and only do bones remain to let them perform laboratory analyses. Bone is the most resistant tissue in animal body to time depending degradation and putrefaction, but it is often hard to extract DNA from it because of its highly mineralized structure, which makes DNA extraction and/or amplification hard to carry out. We have performed human nuclear DNA extraction and STR typing in three different cases, on bones and bone fragments from long time dead persons found buried, in the sea, almost completely burnt and whose tissues were already decomposed. We report these caseworks as we would like to show how forensic scientists are improving their skill in identifying people whose corps have undergone several kinds of processes, even independently on the time passed and the level of putrefaction of their tissues.

  15. Histopathological study and F1L gene sequence analysis of contagious ecthyma in small ruminants of Shiraz suburb, Iran.

    Science.gov (United States)

    Davari, S A; Sayyari, M; Mohammadi, A

    2015-06-01

    Contagious ecthyma, also known as Orf, is a common viral skin disease of sheep and goats caused by a Parapoxvirus. This research was conducted with the aims of histopathological study and genetic analysis of Orf virus with PCR technique based on F1L gene in 50 sheep and goats suspicious of contagious ecthyma in affected areas of Shiraz suburb. All 50 contagious ecthyma-like tissue samples were maintained in 10% buffered formalin, embedded in paraffin, sectioned into 5μm slices and dyed with hematoxylin-eosine. The histopathological examination showed 100% positivity. Epidermal hyperplasia with prominent rete ridges, hydropic degeneration of the necrotic keratinocytes, eosinophilic intracytoplasmic inclusion bodies in vacuolated cells and subcorneal pustules were the main hallmarks of this disease. For molecular analysis, after DNA extraction, all samples were amplified by PCR method and the outcome demonstrated positivity in 25 specimens (50%). Of these, 10 definitely positive specimens were analyzed for nucleotide sequencing. Thus, a strain named Orf-059-Shiraz was recorded in the GeneBank and subsequently underwent phylogenetic analysis. The outcome of the molecular study approved half of the positive ecthyma specimens in histopathological method. Furthermore, phylogenetic analysis based on 059 gene showed that this gene is highly conserved. Utilization of histopathology and clinical signs can assist with rapid and low-cost diagnosis of infectious ecthyma whereas PCR is able to dissociate from similar diseases among clinical samples of endemic regions.

  16. Histopathology of Conjunctivochalasis Compared to Normal Conjunctiva

    Science.gov (United States)

    Hashemian, Hesam; Mahbod, Mirgholamreza; Amoli, Fahimeh A.; Kiarudi, Mohammad Y.; Jabbarvand, Mahmoud; Kheirkhah, Ahmad

    2016-01-01

    Purpose: To evaluate the histopathologic changes in the conjunctiva of patients with conjunctivochalasis (CCh) compared to age-matched controls. Methods: This cross-sectional, controlled study included 27 eyes of 27 patients with CCh and 16 eyes of 16 age-matched controls. A biopsy of the bulbar conjunctiva was performed along the temporal lower lid margin before cataract surgery in both groups. Histopathologic evaluation of the specimens was done with light microscopy using staining with hematoxylin/eosin, periodic acid Schiff, and van Gieson elastic stain. Various histopathologic features of the conjunctival epithelium and stroma were compared between the two groups. Results: The mean age of patients was 62.4 ± 6.9 years in the CCh group and 65.1 ± 6.3 years in the control group (P = 0.54). No significant differences were noted between the two groups in terms of conjunctival epithelial changes including papillomatosis, epithelial clefts, epithelial goblet cells, or infiltration of inflammatory cells. Mean thickness of the conjunctival stroma was 0.21 ± 0.08 mm in the CCh group and 0.26 ± 0.21 mm in the control group (P = 0.10). For the conjunctival stroma, there were no significant differences between the two groups in terms of elastosis, fibrosis, lymphangiectasia, or infiltration of inflammatory cells. Conclusion: No noticeable differences were found in the histopathologic features by light microscopy between eyes with CCh and those of age-matched controls. Therefore, the primary pathology of CCh may not be within the conjunctiva itself. Instead, loose attachment of the conjunctiva to the underlying tissue may be the reason for the redundant folds in the bulbar conjunctiva. PMID:27994801

  17. Radiological Features and Postoperative Histopathologic Diagnosis ...

    African Journals Online (AJOL)

    Radiological Features and Postoperative Histopathologic Diagnosis of Intracranial Masses at Tikur Anbessa ... East and Central African Journal of Surgery ... features and postoperative histopathology diagnosis of intracranial mass lesions.

  18. Bisulfite-Based DNA Methylation Analysis from Recent and Archived Formalin-Fixed, Paraffin Embedded Colorectal Tissue Samples.

    Science.gov (United States)

    Kalmár, Alexandra; Péterfia, Bálint; Hollósi, Péter; Wichmann, Barnabás; Bodor, András; Patai, Árpád V; Schöller, Andrea; Krenács, Tibor; Tulassay, Zsolt; Molnár, Béla

    2015-09-01

    We aimed to test the applicability of formalin-fixed and paraffin-embedded (FFPE) tissue samples for gene specific DNA methylation analysis after using two commercially available DNA isolation kits. Genomic DNA was isolated from 5 colorectal adenocarcinomas and 5 normal adjacent tissues from "recent", collected within 6 months, and "archived", collected more than 5 years ago, FFPE tissues using either High Pure FFPET DNA Isolation kit or QIAamp DNA FFPE Tissue kit. DNA methylation analysis of MAL, SFRP1 and SFRP2 genes, known to be hypermethylated in CRC, was performed using methylation-sensitive high resolution melting (MS-HRM) analysis and sequencing. QIAamp (Q) method resulted in slightly higher recovery in archived (HP: 1.22 ± 3.18 μg DNA; Q: 3.00 ± 4.04 μg DNA) and significantly (p < 0.05) higher recovery in recent samples compared to High Pure method (HP) (HP: 4.10 ± 2.91 μg DNA; Q: 11.51 ± 7.50 μg DNA). Both OD260/280 and OD260/230 ratios were lower, but still high in the High Pure isolated archived and recent samples compared to those isolated with QIAamp. Identical DNA methylation patterns were detected for all 3 genes tested by MS-HRM with both isolation kits in the recent group. However, despite of higher DNA recovery in QIAamp slightly more reproducible methylation results were obtained from High Pure isolated archived samples. Sequencing confirmed DNA hypermethylation in CRCs. In conclusion, reproducible DNA methylation patterns were obtained from recent samples using both isolation kits. However, long term storage may affect the reliability of the results leading to moderate differences between the efficiency of isolation kits.

  19. Identification of immune cell infiltration in hematoxylin-eosin stained breast cancer samples: texture-based classification of tissue morphologies

    Science.gov (United States)

    Turkki, Riku; Linder, Nina; Kovanen, Panu E.; Pellinen, Teijo; Lundin, Johan

    2016-03-01

    The characteristics of immune cells in the tumor microenvironment of breast cancer capture clinically important information. Despite the heterogeneity of tumor-infiltrating immune cells, it has been shown that the degree of infiltration assessed by visual evaluation of hematoxylin-eosin (H and E) stained samples has prognostic and possibly predictive value. However, quantification of the infiltration in H and E-stained tissue samples is currently dependent on visual scoring by an expert. Computer vision enables automated characterization of the components of the tumor microenvironment, and texture-based methods have successfully been used to discriminate between different tissue morphologies and cell phenotypes. In this study, we evaluate whether local binary pattern texture features with superpixel segmentation and classification with support vector machine can be utilized to identify immune cell infiltration in H and E-stained breast cancer samples. Guided with the pan-leukocyte CD45 marker, we annotated training and test sets from 20 primary breast cancer samples. In the training set of arbitrary sized image regions (n=1,116) a 3-fold cross-validation resulted in 98% accuracy and an area under the receiver-operating characteristic curve (AUC) of 0.98 to discriminate between immune cell -rich and - poor areas. In the test set (n=204), we achieved an accuracy of 96% and AUC of 0.99 to label cropped tissue regions correctly into immune cell -rich and -poor categories. The obtained results demonstrate strong discrimination between immune cell -rich and -poor tissue morphologies. The proposed method can provide a quantitative measurement of the degree of immune cell infiltration and applied to digitally scanned H and E-stained breast cancer samples for diagnostic purposes.

  20. Histopathological study of parasitic infestation of skin and gill on Oscar (Astronotus ocellatus and discus (Symphysodon discus

    Directory of Open Access Journals (Sweden)

    Forough Mohammadi

    2012-06-01

    Full Text Available Histopathology provides a rapid method to detect effects of irritants and pathogens in different organs and it can be considered as theindicator for abnormal condition for fish environment. The present study was initiated to record the histopathological lesions of gill and skinassociated with external parasites in two common aquarium fish, Oscar (Astronotus ocellatus and Discus (Symphysodon discus fish. TwentyOscar and twenty Discus were obtained from aquarium shops and wet mount was prepared from skin and gill mucosa and histopathologicalstudy was performed on tissue samples of gills and skin on tissue sections which were stained with haematoxilin-eosin. Based on the results,Dactylogyrus sp. was the most prevalent parasites in Oscar and Discus fish. Ichthyophthirius multifliis, Trichodina sp. and Gyrodactylus sp.,Epistylis sp. and Vorticella sp. were seen in the skin and gill of fish. In histopathological examination, fusion of secondary lamella associatedwith hyperplasia, aneurysm, edema, purulent bronchitis were seen. Sections of Ichthyobodo sp. and purulent bronchitis are rare and in skin.Dermatitis was observed. Histopathological lesions in Oscar were in high rate in comparison with lesions which were seen in Discus and theyare in relation to parasitic infestation.

  1. Lack of mutation-histopathology correlation in a patient with Proteus syndrome.

    Science.gov (United States)

    Doucet, Meggie E; Bloomhardt, Hadley M; Moroz, Krzysztof; Lindhurst, Marjorie J; Biesecker, Leslie G

    2016-06-01

    Proteus syndrome (PS) is characterized by progressive, disproportionate, segmental overgrowth, and tumor susceptibility caused by a somatic mosaic AKT1 activating mutation. Each individual has unique manifestations making this disorder extremely heterogeneous. We correlated three variables in 38 tissue samples from a patient who died with PS: the gross affection status, the microscopic affection status, and the mutation level. The AKT1 mutation was measured using a PCR-based RFLP assay. Thirteen samples were grossly normal; six had detectable mutation (2-29%) although four of these six were histopathologically normal. Of the seven grossly normal samples that had no mutation, only four were histologically normal. The mutation level in the grossly abnormal samples was 3-35% and all but the right and left kidneys, skull, and left knee bone, with mutation levels of 19%, 15%, 26%, and 17%, respectively, had abnormal histopathology. The highest mutation level was in a toe bone sample whereas the lowest levels were in the soft tissue surrounding that toe, and an omental fat nodule. We also show here that PS overgrowth can be caused by cellular proliferation or by extracellular matrix expansion. Additionally, papillary thyroid carcinoma was identified, a tumor not previously associated with PS. We conclude that gross pathology and histopathology correlate poorly with mutation levels in PS, that overgrowth can be mediated by cellular proliferation or extracellular matrix expansion, and that papillary thyroid carcinoma is part of the tumor susceptibility of PS. New methods need to be developed to facilitate genotype-phenotype correlation in mosaic disorders. © 2016 Wiley Periodicals, Inc.

  2. HISTOPATHOLOGICAL CHARACTERISTICS OF LYMPH NODE SINUS CONTAINING BLOOD

    Institute of Scientific and Technical Information of China (English)

    YIN Tong; JI Xiao-long

    2001-01-01

    This study is to find out the histopathological characteristics of lymph node sinus containing blood. Routine autopsy was carried out in the randomly selected 102 patients(among them,100 patients died of various diseases, and 2 of non-diseased causes),their superficial lymph nodes locating in the bilateral neck, axilla, inguina, thorax and abdomen were sampled. Haematoxylin-eosin staining was performed on 10% formalin-fixed and paraffin-embedded lymph node tissue sections(5μm).The histological characteristics of the lymph node sinuses containing blood were observed under light microscope. Among the 1362 lymph nodes sampled from the 100 autopsies, lymph sinuses containing blood were found in 809 lymph nodes sampled from 91 cases, but couldn't be seen in the lymph nodes sampled from the non-diseased cases. According to histology, five kinds of lymph sinuses containing blood were found:vascular-opening sinus, blood-deficient sinus, erythrophago-sinus, blood-abundant sinus, and vascular-formative sinus. It is concluded that in the state of disease, the phenomenon of blood in the lymph sinus is not uncommon. Blood could possibly enter into lymph sinus through lymphatic-venous communications between the veins and sinuses in the node. Lymph circulation and blood circulation could communicate with each other in the lymph sinus.

  3. 3-Dimensional quantitative detection of nanoparticle content in biological tissue samples after local cancer treatment

    Energy Technology Data Exchange (ETDEWEB)

    Rahn, Helene, E-mail: helene.rahn@gmail.com [Institute of Fluid Mechanics, Chair of Magnetofluiddynamics, Technische Universitaet Dresden, Dresden 01069 (Germany); Alexiou, Christoph [ENT-Department, Section for Experimental Oncology and Nanomedicine (Else Kröner-Fresenius-Stiftungsprofessur), University Hospital Erlangen, Waldstraße 1, Erlangen 91054 (Germany); Trahms, Lutz [Physikalisch-Technische Bundesanstalt, Abbestraße 2-12, Berlin 10587 (Germany); Odenbach, Stefan [Institute of Fluid Mechanics, Chair of Magnetofluiddynamics, Technische Universitaet Dresden, Dresden 01069 (Germany)

    2014-06-01

    X-ray computed tomography is nowadays used for a wide range of applications in medicine, science and technology. X-ray microcomputed tomography (XµCT) follows the same principles used for conventional medical CT scanners, but improves the spatial resolution to a few micrometers. We present an example of an application of X-ray microtomography, a study of 3-dimensional biodistribution, as along with the quantification of nanoparticle content in tumoral tissue after minimally invasive cancer therapy. One of these minimal invasive cancer treatments is magnetic drug targeting, where the magnetic nanoparticles are used as controllable drug carriers. The quantification is based on a calibration of the XµCT-equipment. The developed calibration procedure of the X-ray-µCT-equipment is based on a phantom system which allows the discrimination between the various gray values of the data set. These phantoms consist of a biological tissue substitute and magnetic nanoparticles. The phantoms have been studied with XµCT and have been examined magnetically. The obtained gray values and nanoparticle concentration lead to a calibration curve. This curve can be applied to tomographic data sets. Accordingly, this calibration enables a voxel-wise assignment of gray values in the digital tomographic data set to nanoparticle content. Thus, the calibration procedure enables a 3-dimensional study of nanoparticle distribution as well as concentration. - Highlights: • Local cancer treatments are promising in reducing negative side effects occurring during conventional chemotherapy. • The nanoparticles play an important role in delivering drugs to the designated area during local cancer treatments as magnetic drug targeting. • We study the nanoparticles distribution in tumor tissue after magnetic drug targeting with X-ray computed tomography. • We achieved a 3-dimensional quantification of the nanoparticles content in tumor tissue out of digital tomographic data.

  4. Revised histopathological consensus classification of joint implant related pathology.

    Science.gov (United States)

    Krenn, V; Morawietz, L; Perino, G; Kienapfel, H; Ascherl, R; Hassenpflug, G J; Thomsen, M; Thomas, P; Huber, M; Kendoff, D; Baumhoer, D; Krukemeyer, M G; Natu, S; Boettner, F; Zustin, J; Kölbel, B; Rüther, W; Kretzer, J P; Tiemann, A; Trampuz, A; Frommelt, L; Tichilow, R; Söder, S; Müller, S; Parvizi, J; Illgner, U; Gehrke, T

    2014-12-01

    This extended classification of joint implant related pathology is a practical histopathologic classification based on defined morphological criteria covering the complete spectrum of pathohistologic changes in periprosthetic tissues. These changes may occur as a consequence of endoprosthetic replacement of large joints and may lead to a reduction in the prosthesis survival rate. We describe the established consensus classification of the periprosthetic membrane, in which aseptic and septic prosthetic loosening can be subdivided into four histological types, as well as histopathological criteria for additional significant pathologies including endoprosthetic-associated arthrofibrosis, particle-induced immunological, inflammatory and toxic mechanisms (adverse reactions), and bone tissue pathologies. These characteristic tissue alterations and their relationships are summarized in the extended classification. Since particle heterogeneity in periprosthetic tissue is high and particle identification is a necessary part of diagnosis, the identification of different types of particles is described in the histopathological particle algorithm. The morphological qualities of prosthetic material particles and the demarcation between abrasion and non-abrasion endogenous particles are also summarized. This feasible classification which is based on low cost standard tissue processing and examination and on well-defined diagnostic criteria is a solid platform for the histological diagnosis of implant associated pathologies providing a stable and reproducible tool for the surgical pathologist. Since this classification is suitable for standardized histopathological diagnostics, it might also provide a useful data set for joint arthroplasty registers, particularly for registers based on so-called routine data.

  5. Decreased Expression of Inhibitor of Caspase-Activated DNase (ICAD) in Renal Cell Carcinoma - Tissue Microarray of Human Samples.

    Science.gov (United States)

    Rajandram, Retnagowri; Razack, Azad H A; Ng, Keng Lim; Gobe, Glenda C

    2016-01-01

    Although primary localised tumours of renal cell carcinoma (RCC) can be treated relatively successfully with surgery, metastatic RCC has poor prognosis because of late diagnosis and resistance to therapies. In the present study, we were interested in profiling the protein expression of "inhibitor of caspase-activated DNase" (ICAD), an apoptosis inhibitor, in kidney cancer and its paired normal kidney. Immunohistochemistry with automated batch staining and morphometry using digital pathology were used to compare ICAD in 121 RCC specimens with their paired normal kidney tissue. Tissue microarray of formalin-fixed, paraffin-embedded archival tissue was used. Intensity and localisation of ICAD were compared between normal and cancer samples, and against grading within the cancers. The results demonstrated that, in this cohort, ICAD was highly expressed in the proximal tubular epithelium of normal kidney, and significantly decreased in clear cell RCC tissue (p < 0.05) as well as other subtypes of RCC (p < 0.01) compared with normal kidney. There was a tendency towards nuclear localisation of ICAD in clear cell RCC, but not in other subtypes of RCC. No significant association was found between ICAD intensity and grade of RCC. In summary, down-regulation of ICAD occurs in RCC. ICAD normally inhibits DNA fragmentation and apoptosis; thus, its down-regulation was unexpected in a cancer known for its resistance to apoptosis. However, these RCC samples were from primary, not metastatic, RCC sites, and down-regulated ICAD may be part of a progressive pathway that promotes RCC metastasis.

  6. Epo receptors are not detectable in primary human tumor tissue samples.

    Directory of Open Access Journals (Sweden)

    Steve Elliott

    Full Text Available Erythropoietin (Epo is a cytokine that binds and activates an Epo receptor (EpoR expressed on the surface of erythroid progenitor cells to promote erythropoiesis. While early studies suggested EpoR transcripts were expressed exclusively in the erythroid compartment, low-level EpoR transcripts were detected in nonhematopoietic tissues and tumor cell lines using sensitive RT-PCR methods. However due to the widespread use of nonspecific anti-EpoR antibodies there are conflicting data on EpoR protein expression. In tumor cell lines and normal human tissues examined with a specific and sensitive monoclonal antibody to human EpoR (A82, little/no EpoR protein was detected and it was not functional. In contrast, EpoR protein was reportedly detectable in a breast tumor cell line (MCF-7 and breast cancer tissues with an anti-EpoR polyclonal antibody (M-20, and functional responses to rHuEpo were reported with MCF-7 cells. In another study, a functional response was reported with the lung tumor cell line (NCI-H838 at physiological levels of rHuEpo. However, the specificity of M-20 is in question and the absence of appropriate negative controls raise questions about possible false-positive effects. Here we show that with A82, no EpoR protein was detectable in normal human and matching cancer tissues from breast, lung, colon, ovary and skin with little/no EpoR in MCF-7 and most other breast and lung tumor cell lines. We show further that M-20 provides false positive staining with tissues and it binds to a non-EpoR protein that migrates at the same size as EpoR with MCF-7 lysates. EpoR protein was detectable with NCI-H838 cells, but no rHuEpo-induced phosphorylation of AKT, STAT3, pS6RP or STAT5 was observed suggesting the EpoR was not functional. Taken together these results raise questions about the hypothesis that most tumors express high levels of functional EpoR protein.

  7. Three-dimensional digital breast histopathology imaging

    Science.gov (United States)

    Clarke, G. M.; Peressotti, C.; Mawdsley, G. E.; Eidt, S.; Ge, M.; Morgan, T.; Zubovits, J. T.; Yaffe, M. J.

    2005-04-01

    We have developed a digital histology imaging system that has the potential to improve the accuracy of surgical margin assessment in the treatment of breast cancer by providing finer sampling and 3D visualization. The system is capable of producing a 3D representation of histopathology from an entire lumpectomy specimen. We acquire digital photomicrographs of a stack of large (120 x 170 mm) histology slides cut serially through the entire specimen. The images are then registered and displayed in 2D and 3D. This approach dramatically improves sampling and can improve visualization of tissue structures compared to current, small-format histology. The system consists of a brightfield microscope, adapted with a freeze-frame digital video camera and a large, motorized translation stage. The image of each slide is acquired as a mosaic of adjacent tiles, each tile representing one field-of-view of the microscope, and the mosaic is assembled into a seamless composite image. The assembly is done by a program developed to build image sets at six different levels within a multiresolution pyramid. A database-linked viewing program has been created to efficiently register and display the animated stack of images, which occupies about 80 GB of disk space per lumpectomy at full resolution, on a high-resolution (3840 x 2400 pixels) colour monitor. The scanning or tiling approach to digitization is inherently susceptible to two artefacts which disrupt the composite image, and which impose more stringent requirements on system performance. Although non-uniform illumination across any one isolated tile may not be discernible, the eye readily detects this non-uniformity when the entire assembly of tiles is viewed. The pattern is caused by deficiencies in optical alignment, spectrum of the light source, or camera corrections. The imaging task requires that features as small as 3.2 &mum in extent be seamlessly preserved. However, inadequate accuracy in positioning of the translation

  8. Detection of Jaagsiekte sheep retrovirus in apparently healthy sheep by real-time TaqMan PCR in comparison with histopathological findings

    Directory of Open Access Journals (Sweden)

    Bahari Aliasghar

    2016-03-01

    Full Text Available Introduction: The aim of this study was to use TaqMan real-time PCR technique to investigate Jaagsiekte sheep retrovirus (JSRV proviral DNA in whole blood samples of sheep, and compare the results to those of histopathological examinations. Material and Methods: Eighty blood samples from clinically healthy sheep were randomly collected before the animals were slaughtered. Ten tissue samples from each lung and associated caudal mediastinal lymph node were taken. Results: Fifteen (18.75% blood samples were found to contain proviral DNA, and 11 (13.75% corresponding lung samples showed microscopic changes consistent with ovine pulmonary adenocarcinoma. None of the samples displayed metastases to the caudal mediastinal lymph nodes. The prominent pattern of neoplastic nodules consisted of acinar (alveolar form. Conclusion: The results indicated the higher sensitivity of real-time PCR compared to histopathological examinations in detection of ovine pulmonary adenocarcinoma.

  9. A new liquid chromatography-tandem mass spectrometry method for determination of parabens in human placental tissue samples.

    Science.gov (United States)

    Jiménez-Díaz, I; Vela-Soria, F; Zafra-Gómez, A; Navalón, A; Ballesteros, O; Navea, N; Fernández, M F; Olea, N; Vílchez, J L

    2011-05-15

    Endocrine disruptors are a group of organic compounds widely used, which are ubiquitous in the environment and in biological samples. The main effect of these compounds is associated with their ability to mimic or block the action of natural hormones in living organisms, including humans. Parabens (esters of p-hydroxybenzoic acid) belong to this group of compounds. In this work, we propose a new liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to asses the presence of parabens most commonly used in industrial applications (methyl-, ethyl-, propyl- and butyl-paraben) in samples of human placental tissue. The method involves the extraction of the analytes from the samples using ethyl acetate, followed by a clean-up step using centrifugation prior to their quantification by LC-MS/MS using an atmospheric pressure chemical ionization (APCI) interface in the negative mode. Deuterated bisphenol A (BPA-d(16)) was used as surrogate. Found detection limits (LOD) ranged from 0.03 to 0.06 ng g(-1) and quantification limits (LOQ) from 0.1 to 0.2 ng g(-1), while inter- and intra-day variability was under 13.8%. The method was validated using standard addition calibration and a spike recovery assay. Recovery rates for spiked samples ranged from 82% to 108%. This method was satisfactorily applied for the determination of parabens in 50 placental tissue samples collected from women who live in the province of Granada (Spain).

  10. Clinical Use of Fungal PCR from Deep Tissue Samples in the Diagnosis of Invasive Fungal Diseases - A Retrospective, Observational Study.

    Science.gov (United States)

    Ala-Houhala, Mari; Koukila-Kähkölä, Pirkko; Antikainen, Jenni; Valve, Jaana; Kirveskari, Juha; Anttila, Veli-Jukka

    2017-09-01

    We evaluated the clinical use of panfungal polymerase chain reaction (PCR) for diagnosis of invasive fungal diseases (IFDs). We focused on the deep tissue samples. We first described the design of panfungal PCR, which is in clinical use at Helsinki University Hospital. Secondly, we retrospectively evaluated the results of 307 fungal PCR tests performed from 2013-2015. Samples were taken from normally sterile tissues and fluids. The patient population was non-selected. We classified the likelihood of IFD according to the criteria of the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG), compering the fungal PCR results to the likelihood of IFD along with the culture and microscopy results. There were 48 (16%) positive and 259 (84%) negative PCR results. The sensitivity and specificity of PCR for diagnosing invasive fungal disease were 60.5% and 91.7%, respectively, while the NPV and PPV were 93.4% and 54.2%, respectively. The concordance between the PCR and the culture results was 86% and 87% between PCR and microscopy, respectively. Of the 48 patients with positive PCR results 23 had a proven or probable IFD. Fungal PCR can be useful for diagnosing IFDs in deep tissue samples. It is beneficial to combine fungal PCR with culture and microscopy. Copyright © 2017. Published by Elsevier Ltd.

  11. Sampling

    CERN Document Server

    Thompson, Steven K

    2012-01-01

    Praise for the Second Edition "This book has never had a competitor. It is the only book that takes a broad approach to sampling . . . any good personal statistics library should include a copy of this book." —Technometrics "Well-written . . . an excellent book on an important subject. Highly recommended." —Choice "An ideal reference for scientific researchers and other professionals who use sampling." —Zentralblatt Math Features new developments in the field combined with all aspects of obtaining, interpreting, and using sample data Sampling provides an up-to-date treat

  12. Histopathological characterization of a Cameron lesion.

    Science.gov (United States)

    Katz, Jordan; Brar, Sonia; Sidhu, Jagmohan S

    2012-10-01

    Cameron lesions are linear erosions located at the neck of a hiatal hernia (HH) in patients with a large HH. The prevalence has been seen in up to 5% of patients with HH who undergo esophagogastroduodenoscopy, and they can be associated with overt gastrointestinal bleeding or anemia. These lesions occur due to vascular compression by the diaphragm in a large sliding HH. Histopathologic changes seen in the biopsy tissue of a Cameron lesion are due to ischemia, but this ischemia is reversible with treatment of HH. The existence of this entity and the histopathologic picture of a Cameron lesion is not well known to pathologists, and therefore, a microscopic picture of a Cameron lesion can be easily confused with ischemic gastritis. Ischemic gastritis is the result of atherosclerosis, usually seen in older people, unrelated to HH, and is not easily reversible. The authors received a gastric biopsy of a hiatal hernia without any associated clinical diagnosis of a Cameron lesion conveyed to the pathologist. This biopsy tissue showed ischemic changes in the gastric mucosa on microscopic examination. Diagnosis of ischemic gastritis was considered but ruled out after the case was discussed with the gastroenterologist. The correct diagnosis was made once the clinical diagnosis of HH with Cameron lesion (ie, a vertical red erosion) was made known to the pathologist. By reporting this case, the authors aim to increase awareness of Cameron lesion among pathologists so that they ask about the presence of a Cameron lesion before making the diagnosis of ischemic gastritis.

  13. A hierarchical Naïve Bayes Model for handling sample heterogeneity in classification problems: an application to tissue microarrays

    Directory of Open Access Journals (Sweden)

    Piergiorgi Paolo

    2006-11-01

    Full Text Available Abstract Background Uncertainty often affects molecular biology experiments and data for different reasons. Heterogeneity of gene or protein expression within the same tumor tissue is an example of biological uncertainty which should be taken into account when molecular markers are used in decision making. Tissue Microarray (TMA experiments allow for large scale profiling of tissue biopsies, investigating protein patterns characterizing specific disease states. TMA studies deal with multiple sampling of the same patient, and therefore with multiple measurements of same protein target, to account for possible biological heterogeneity. The aim of this paper is to provide and validate a classification model taking into consideration the uncertainty associated with measuring replicate samples. Results We propose an extension of the well-known Naïve Bayes classifier, which accounts for biological heterogeneity in a probabilistic framework, relying on Bayesian hierarchical models. The model, which can be efficiently learned from the training dataset, exploits a closed-form of classification equation, thus providing no additional computational cost with respect to the standard Naïve Bayes classifier. We validated the approach on several simulated datasets comparing its performances with the Naïve Bayes classifier. Moreover, we demonstrated that explicitly dealing with heterogeneity can improve classification accuracy on a TMA prostate cancer dataset. Conclusion The proposed Hierarchical Naïve Bayes classifier can be conveniently applied in problems where within sample heterogeneity must be taken into account, such as TMA experiments and biological contexts where several measurements (replicates are available for the same biological sample. The performance of the new approach is better than the standard Naïve Bayes model, in particular when the within sample heterogeneity is different in the different classes.

  14. Comparing paraffined and deparaffinized breast cancer tissue samples and an analysis of Raman spectroscopy and infrared methods

    Science.gov (United States)

    Depciuch, J.; Kaznowska, E.; Szmuc, K.; Zawlik, I.; Cholewa, M.; Heraud, P.; Cebulski, J.

    2016-05-01

    Breast cancer makes up a quarter of all cancer in women, which is why research into new diagnostic methods and sample preparations need to be developed at an accelerated pace. Researchers are looking for diagnostic tools to detect when an individual has cancer cells and use that information to see what measurements and approaches can be used to take further diagnostic steps. The most common method of sample preparation is the imbibing of tumor tissue in paraffin, which can produce a background for spectroscopic measurements in the range of 500-3500 cm-1. In this study we demonstrated that proper preparation of paraffin-embedded specimens and the measurement methodology can eliminate paraffin vibration, as was done in the work Depciuch et al. 2015. Thanks to this spectroscopic technique there may become a reliable and accurate method of diagnosing breast cancer based on the evidence found from the prepared samples. The study compared the results obtained through Raman spectroscopy and FTIR (Fourier Transform Infrared) measurements of healthy and cancerous breast tissues that were either embedded in paraffin or deparaffinized. The resulting spectrum and accurate analysis led to the conclusion that the appropriate measurement of the background and the elimination of peaks from the paraffin had the greatest impact on the reliability of results. Furthermore, after the accurate, detailed studies FTIR and Raman spectroscopy on samples of breast tissue that were deparaffinized or embedded in paraffin, including a complete analysis of the peak after transformation Kramers-Kröning (KK), it was found that sample preparation did not affect the result obtained by measuring the reflectance in the mid-infrared range, and that this only had a minimal effect relating to the intensity obtained by the measurement of the Raman peak. Only in special cases, when Raman spectroscopic methods are used for research to find the peculiarities of the spectra, are deparaffinization recommended

  15. Whole-genome gene expression profiling of formalin-fixed, paraffin-embedded tissue samples.

    Directory of Open Access Journals (Sweden)

    Craig April

    Full Text Available BACKGROUND: We have developed a gene expression assay (Whole-Genome DASL, capable of generating whole-genome gene expression profiles from degraded samples such as formalin-fixed, paraffin-embedded (FFPE specimens. METHODOLOGY/PRINCIPAL FINDINGS: We demonstrated a similar level of sensitivity in gene detection between matched fresh-frozen (FF and FFPE samples, with the number and overlap of probes detected in the FFPE samples being approximately 88% and 95% of that in the corresponding FF samples, respectively; 74% of the differentially expressed probes overlapped between the FF and FFPE pairs. The WG-DASL assay is also able to detect 1.3-1.5 and 1.5-2 -fold changes in intact and FFPE samples, respectively. The dynamic range for the assay is approximately 3 logs. Comparing the WG-DASL assay with an in vitro transcription-based labeling method yielded fold-change correlations of R(2 approximately 0.83, while fold-change comparisons with quantitative RT-PCR assays yielded R(2 approximately 0.86 and R(2 approximately 0.55 for intact and FFPE samples, respectively. Additionally, the WG-DASL assay yielded high self-correlations (R(2>0.98 with low intact RNA inputs ranging from 1 ng to 100 ng; reproducible expression profiles were also obtained with 250 pg total RNA (R(2 approximately 0.92, with approximately 71% of the probes detected in 100 ng total RNA also detected at the 250 pg level. When FFPE samples were assayed, 1 ng total RNA yielded self-correlations of R(2 approximately 0.80, while still maintaining a correlation of R(2 approximately 0.75 with standard FFPE inputs (200 ng. CONCLUSIONS/SIGNIFICANCE: Taken together, these results show that WG-DASL assay provides a reliable platform for genome-wide expression profiling in archived materials. It also possesses utility within clinical settings where only limited quantities of samples may be available (e.g. microdissected material or when minimally invasive procedures are performed (e

  16. Histopathologic Overlap between Fibrosing Mediastinitis and IgG4-Related Disease

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    Tobias Peikert

    2012-01-01

    Full Text Available Fibrosing mediastinitis (FM and IgG4-related disease (IgG4-RD are two fibroinflammatory disorders with potentially overlapping clinical and radiological features. In this paper, we looked for histopathologic features of IgG4-RD and enumerated infiltrating IgG4-positive plasma cells within mediastinal tissue biopsies from FM patients. We identified 15 consecutive FM surgical mediastinal tissue biopsies between 1985 and 2006. All patients satisfied the clinical and radiological diagnostic criteria for FM. All patients had either serological or radiological evidence of prior histoplasmosis or granulomatous disease, respectively. Formalin-fixed paraffin-embedded tissue sections of all patients were stained for H&E, IgG, and IgG4. Three samples met the predefined diagnostic criteria for IgG4-RD. In addition, characteristic histopathologic changes of IgG4-RD in the absence of diagnostic numbers of tissue infiltrating IgG4-positive plasma cells were seen in a number of additional cases (storiform cell-rich fibrosis in 11 cases, lymphoplasmacytic infiltrate in 7 cases, and obliterative phlebitis/arteritis in 2 cases. We conclude that up to one-third of histoplasmosis or granulomatous-disease-associated FM cases demonstrate histopathological features of IgG4-RD spectrum. Whether these changes occur as the host immune response against Histoplasma or represent a manifestation of IgG4-RD remains to be determined. Studies to prospectively identify these cases and evaluate their therapeutic responses to glucocorticoids and/or other immunosuppressive agents such as rituximab are warranted.

  17. Determination of protein carbonyls in plasma, cell extracts, tissue homogenates, isolated proteins: Focus on sample preparation and derivatization conditions.

    Science.gov (United States)

    Weber, Daniela; Davies, Michael J; Grune, Tilman

    2015-08-01

    Protein oxidation is involved in regulatory physiological events as well as in damage to tissues and is thought to play a key role in the pathophysiology of diseases and in the aging process. Protein-bound carbonyls represent a marker of global protein oxidation, as they are generated by multiple different reactive oxygen species in blood, tissues and cells. Sample preparation and stabilization are key steps in the accurate quantification of oxidation-related products and examination of physiological/pathological processes. This review therefore focuses on the sample preparation processes used in the most relevant methods to detect protein carbonyls after derivatization with 2,4-dinitrophenylhydrazine with an emphasis on measurement in plasma, cells, organ homogenates, isolated proteins and organelles. Sample preparation, derivatization conditions and protein handling are presented for the spectrophotometric and HPLC method as well as for immunoblotting and ELISA. An extensive overview covering these methods in previously published articles is given for researchers who plan to measure protein carbonyls in different samples.

  18. The presence of enterovirus, adenovirus, and parvovirus B19 in myocardial tissue samples from autopsies

    DEFF Research Database (Denmark)

    Nielsen, Trine Skov; Hansen, Jakob; Nielsen, Lars Peter

    2014-01-01

    of adenovirus, enterovirus, and parvovirus B19 (PVB) in myocardial autopsy samples from myocarditis related deaths and in non-inflamed control hearts in an effort to clarify their significance as the causes of myocarditis in a forensic material. METHODS: We collected all autopsy cases diagnosed with myocarditis...

  19. Attitudes towards biomedical use of tissue sample collections, consent, and biobanks among Finns

    DEFF Research Database (Denmark)

    Tupasela, Aaro Mikael; Sihvo, Sinikka; Snell, Karolna

    2010-01-01

    To ascertain the attitudes towards the use of existing diagnostic and research samples, the setting up of a national biobank, and different types of informed consent among Finns. Method: A population survey of 2,400 randomly selected Finns aged 24-65 was conducted at the beginning of 2007....

  20. MicroRNA Stability in Postmortem FFPE Tissues: Quantitative Analysis Using Autoptic Samples from Acute Myocardial Infarction Patients.

    Science.gov (United States)

    Kakimoto, Yu; Kamiguchi, Hiroshi; Ochiai, Eriko; Satoh, Fumiko; Osawa, Motoki

    2015-01-01

    MicroRNAs (miRNAs) are very short (18-24 nucleotides) nucleic acids that are expressed in a number of biological tissues and have been shown to be more resistant to extreme temperatures and pH compared to longer RNA molecules, like mRNAs. As miRNAs contribute to diverse biological process and respond to various kinds of cellular stress, their utility as diagnostic biomarkers and/or therapeutic targets has recently been explored. Here, we have evaluated the usefulness of miRNA quantification during postmortem examination of cardiac tissue from acute myocardial infarction (AMI) patients. Cardiac tissue was collected within one week of the patient's death and either frozen (19 samples) or fixed in formalin for up to three years (36 samples). RNA integrity was evaluated with an electropherogram, and it appears that longer RNAs are fragmented after death in the long-term fixed samples. Quantitative PCR was also performed for seven miRNAs and three other small RNAs in order to determine the appropriate controls for our postmortem analysis. Our data indicate that miR-191 and miR-26b are more suitable than the other types of small RNA molecules as they are stably detected after death and long-term fixation. Further, we also applied our quantitation method, using these endogenous controls, to evaluate the expression of three previously identified miRNA biomarkers, miR-1, miR-208b, and miR-499a, in formalin-fixed tissues from AMI patients. Although miR-1 and miR-208b decreased (1.4-fold) and increased (1.2-fold), respectively, in the AMI samples compared to the controls, the significance of these changes was limited by our sample size. In contrast, the relative level of miR-499a was significantly decreased in the AMI samples (2.1-fold). This study highlights the stability of miRNAs after death and long-term fixation, validating their use as reliable biomarkers for AMI during postmortem examination.

  1. MicroRNA Stability in Postmortem FFPE Tissues: Quantitative Analysis Using Autoptic Samples from Acute Myocardial Infarction Patients.

    Directory of Open Access Journals (Sweden)

    Yu Kakimoto

    Full Text Available MicroRNAs (miRNAs are very short (18-24 nucleotides nucleic acids that are expressed in a number of biological tissues and have been shown to be more resistant to extreme temperatures and pH compared to longer RNA molecules, like mRNAs. As miRNAs contribute to diverse biological process and respond to various kinds of cellular stress, their utility as diagnostic biomarkers and/or therapeutic targets has recently been explored. Here, we have evaluated the usefulness of miRNA quantification during postmortem examination of cardiac tissue from acute myocardial infarction (AMI patients. Cardiac tissue was collected within one week of the patient's death and either frozen (19 samples or fixed in formalin for up to three years (36 samples. RNA integrity was evaluated with an electropherogram, and it appears that longer RNAs are fragmented after death in the long-term fixed samples. Quantitative PCR was also performed for seven miRNAs and three other small RNAs in order to determine the appropriate controls for our postmortem analysis. Our data indicate that miR-191 and miR-26b are more suitable than the other types of small RNA molecules as they are stably detected after death and long-term fixation. Further, we also applied our quantitation method, using these endogenous controls, to evaluate the expression of three previously identified miRNA biomarkers, miR-1, miR-208b, and miR-499a, in formalin-fixed tissues from AMI patients. Although miR-1 and miR-208b decreased (1.4-fold and increased (1.2-fold, respectively, in the AMI samples compared to the controls, the significance of these changes was limited by our sample size. In contrast, the relative level of miR-499a was significantly decreased in the AMI samples (2.1-fold. This study highlights the stability of miRNAs after death and long-term fixation, validating their use as reliable biomarkers for AMI during postmortem examination.

  2. Adequately defining tumor cell proportion in tissue samples for molecular testing improves interobserver reproducibility of its assessment.

    Science.gov (United States)

    Lhermitte, Benoît; Egele, Caroline; Weingertner, Noëlle; Ambrosetti, Damien; Dadone, Bérengère; Kubiniek, Valérie; Burel-Vandenbos, Fanny; Coyne, John; Michiels, Jean-François; Chenard, Marie-Pierre; Rouleau, Etienne; Sabourin, Jean-Christophe; Bellocq, Jean-Pierre

    2017-01-01

    Gene mutation status assessment of tumors has become standard practice in diagnostic pathology. This is done using samples comprising tumor cells but also non-tumor cells, which may dramatically dilute the proportion of tumor DNA and induce false negative results. Increasing sensitivity of molecular tests presently allows detection of a targeted mutation in a sample with a small percentage of tumor cells, but assessment of tumor cellularity remains essential to adequately interpret the results of molecular tests. Comprehensive tumor cell counting would provide the most reliable approach but is time consuming, and therefore rough global estimations are used, the reliability of which has been questioned in view of their potential clinical impact. The French association for quality assurance in pathology (AFAQAP) conducted two external quality assurance schemes, partly in partnership with the French group of oncology cytogenomics (GFCO). The purpose of the schemes was to (1) evaluate how tumor cellularity is assessed on tissue samples, (2) identify reasons for discrepancies, and (3) provide recommendations for standardization and improvement. Tumor cell percentages in tissue samples of lung and colon cancer were estimated by 40-50 participants, on 10 H&E virtual slides and 20 H&E conventional slides. The average difference between lowest and highest estimated percentage was 66. This was largely due to inadequate definition of cellularity, reflecting confusion between the percentage of tumor cells and the percentage of the area occupied by tumor in the assessed region. The widest range of interobserver variation was observed for samples with dense or scattered lymphocytic infiltrates or with mucinous stroma. Estimations were more accurate in cases with a low percentage of tumor cells. Macrodissection of the most homogeneous area in the tissue reduced inter-laboratory variation. We developed a rating system indicating potential clinical impact of a discrepancy. Fewer

  3. Histopathological changes in the gills and liver of Prochilodus lineatus from the Salado River basin (Santa Fe, Argentina).

    Science.gov (United States)

    Troncoso, Ileana C; Cazenave, Jimena; Bacchetta, Carla; Bistoni, María de Los Angeles

    2012-06-01

    This study evaluated the histopathological changes in gills and liver of Prochilodus lineatus inhabiting the Salado River basin. Fish were collected in four different sampling stations. The histological lesions in the tissues were examined under light microscopy and evaluated with quantitative analyses. The morphometric analysis of the gills showed a significant shortening of secondary lamellae and a lower percentage of area for gas exchange in fish from station 1 (an urban area, located near the mouth of the Salado River) in comparison with fish gills from the reference site (station 4, a relatively pristine area). Moreover, a significantly higher area occupied with necrotic foci and the occurrence of an important inflammatory response were observed in fish liver of station 1 than the samples caught from other stations. Thus, histopathological evidences showed differences among sites, which could be related to different environmental conditions.

  4. Expression of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinases 1 (TIMP-1) by colorectal cancer cells and adjacent stroma cells--associations with histopathology and patients outcome

    DEFF Research Database (Denmark)

    Jensen, Søren Astrup; Vainer, Ben; Bartels, Annette;

    2010-01-01

    To elucidate cellular features accountable for colorectal cancers' (CRC) capability to invade normal tissue and to metastasize, we investigated the level of the collagenase matrix metalloproteinase 9 (MMP-9) and its physiological inhibitor tissue inhibitor of metalloproteinases 1 (TIMP-1) in cancer...

  5. Expression of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinases 1 (TIMP-1) by colorectal cancer cells and adjacent stroma cells--associations with histopathology and patients outcome

    DEFF Research Database (Denmark)

    Jensen, Søren Astrup; Vainer, Ben; Bartels, Annette

    2010-01-01

    To elucidate cellular features accountable for colorectal cancers' (CRC) capability to invade normal tissue and to metastasize, we investigated the level of the collagenase matrix metalloproteinase 9 (MMP-9) and its physiological inhibitor tissue inhibitor of metalloproteinases 1 (TIMP-1) in cancer...

  6. Expression of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinases 1 (TIMP-1) by colorectal cancer cells and adjacent stroma cells--associations with histopathology and patients outcome

    DEFF Research Database (Denmark)

    Jensen, Søren Astrup; Vainer, Ben; Bartels, Annette

    2010-01-01

    AIM: To elucidate cellular features accountable for colorectal cancers' (CRC) capability to invade normal tissue and to metastasize, we investigated the level of the collagenase matrix metalloproteinase 9 (MMP-9) and its physiological inhibitor tissue inhibitor of metalloproteinases 1 (TIMP-1...

  7. Liver biopsy histopathology for diagnosis of Johne's disease in sheep.

    Science.gov (United States)

    Smith, S L; Wilson, P R; Collett, M G; Heuer, C; West, D M; Stevenson, M; Chambers, J P

    2014-09-01

    Sheep with Johne's disease develop epithelioid macrophage microgranulomas, specific to Mycobacterium avium subsp. paratuberculosis (Map) infection, in the terminal ileum, mesenteric lymph nodes, and organs distant to the alimentary tract such as the liver. The objectives of this study were to determine whether liver pathology was present in ewes affected by Map and whether liver cores provide adequate tissue for this potential diagnostic marker. One hundred and twenty-six adult, low body condition ewes were euthanized, necropsied, and underwent simulated liver biopsy. Ileal lesions typical of Map were found in 60 ewes. Hepatic epithelioid microgranulomas were observed in all ewes with Type 3b (n = 40) and 82% (n = 11) with Type 3c ileal lesions. None were found in ewes unaffected by Map or with Type 1, 2, or 3a ileal lesions. Liver biopsy core samples provided adequate tissue for histopathology with a sensitivity and specificity of 96% (95% confidence interval [CI], 0.87-0.99) and 100% (95% CI, 0.95-1), respectively for detection of types 3b and 3c ileal lesions.

  8. Equine Odontoclastic Tooth Resorption and Hypercementosis: Histopathologic Features.

    Science.gov (United States)

    Smedley, R C; Earley, E T; Galloway, S S; Baratt, R M; Rawlinson, J E

    2015-09-01

    Equine odontoclastic tooth resorption and hypercementosis (EOTRH) is a painful progressive condition of older horses that involves multiple teeth, including canines and incisors. EOTRH is uncommonly recognized by veterinary pathologists and in some cases may be misdiagnosed as cementoblastoma. The cause is unknown. The goals of this study were to describe the histopathologic features of EOTRH in 17 affected horses from the United States and to increase awareness of this condition. Samples ranged from affected tooth to the entire rostral mandible and maxilla. Affected teeth exhibited cemental hyperplasia and lysis. The marked proliferation of cementum in severe cases caused bulbous enlargement of the intra-alveolar portions of affected teeth. Several teeth contained necrotic debris, bacteria, and plant material in the regions of cemental lysis. All horses exhibited dentinal lysis in at least affected tooth, and several contained necrotic debris in these regions. Endodontic disease was often present with inflammation, lysis, necrotic debris, fibrosis, and/or a thin rim of atubular mineralized tissue in the pulp cavity. Periodontal disease was a common feature that was primarily characterized by moderate lymphoplasmacytic inflammation. Resorption with secondary hypercementosis appears to begin on the external surface of the teeth rather than within the pulp cavity. Distinguishing EOTRH from other diseases requires a complete history that includes the number and location of affected teeth, a gross description of regional hard/soft tissue health, and radiographic findings.

  9. Intravascular papillary endothelial hyperplasia: An unusual histopathological entity.

    Science.gov (United States)

    Mahapatra, Qury S; Sahai, Kavita; Malik, Ajay; Mani, N S

    2015-01-01

    Intravascular papillary endothelial hyperplasia (Masson's tumor) is a benign lesion of the skin and subcutaneous tissue consisting of a reactive proliferation of endothelial cells with papillary formations related to a thrombus. It poses a diagnostic challenge as the clinical signs and symptoms are nonspecific and may mimic a soft tissue sarcoma. The diagnosis is based on histopathology. Here we report two cases of Masson's hemangioma occurring on the upper lip and on the left hand.

  10. Intravascular papillary endothelial hyperplasia: An unusual histopathological entity

    Directory of Open Access Journals (Sweden)

    Qury S Mahapatra

    2015-01-01

    Full Text Available Intravascular papillary endothelial hyperplasia (Masson′s tumor is a benign lesion of the skin and subcutaneous tissue consisting of a reactive proliferation of endothelial cells with papillary formations related to a thrombus. It poses a diagnostic challenge as the clinical signs and symptoms are nonspecific and may mimic a soft tissue sarcoma. The diagnosis is based on histopathology. Here we report two cases of Masson′s hemangioma occurring on the upper lip and on the left hand.

  11. Micro-scaled high-throughput digestion of plant tissue samples for multi-elemental analysis

    Directory of Open Access Journals (Sweden)

    Husted Søren

    2009-09-01

    Full Text Available Abstract Background Quantitative multi-elemental analysis by inductively coupled plasma (ICP spectrometry depends on a complete digestion of solid samples. However, fast and thorough sample digestion is a challenging analytical task which constitutes a bottleneck in modern multi-elemental analysis. Additional obstacles may be that sample quantities are limited and elemental concentrations low. In such cases, digestion in small volumes with minimum dilution and contamination is required in order to obtain high accuracy data. Results We have developed a micro-scaled microwave digestion procedure and optimized it for accurate elemental profiling of plant materials (1-20 mg dry weight. A commercially available 64-position rotor with 5 ml disposable glass vials, originally designed for microwave-based parallel organic synthesis, was used as a platform for the digestion. The novel micro-scaled method was successfully validated by the use of various certified reference materials (CRM with matrices rich in starch, lipid or protein. When the micro-scaled digestion procedure was applied on single rice grains or small batches of Arabidopsis seeds (1 mg, corresponding to approximately 50 seeds, the obtained elemental profiles closely matched those obtained by conventional analysis using digestion in large volume vessels. Accumulated elemental contents derived from separate analyses of rice grain fractions (aleurone, embryo and endosperm closely matched the total content obtained by analysis of the whole rice grain. Conclusion A high-throughput micro-scaled method has been developed which enables digestion of small quantities of plant samples for subsequent elemental profiling by ICP-spectrometry. The method constitutes a valuable tool for screening of mutants and transformants. In addition, the method facilitates studies of the distribution of essential trace elements between and within plant organs which is relevant for, e.g., breeding programmes aiming at

  12. CBs levels in deer tissue samples following an accidental release from a special waste treatment center : 2003 results

    Energy Technology Data Exchange (ETDEWEB)

    Mackenzie, A.; Zhang, W.; Gabos, S.; Schopflocher, D.; Robb, J. [Alberta Health and Wellness, Edmonton, AB (Canada). Health Surveillance; Ikonomou, M. [Inst. of Ocean Sciences, Sidney, BC (Canada)

    2005-07-01

    High levels of dioxins and furans (PCDD/Fs) were observed in deer and mouse tissues examined after an accidental release of PCDD/F contaminants in the Swan Hills area of Alberta. This paper provided details of follow-up sampling conducted to assess changes in CB concentrations of whitetail deer and mule deer in the region. Field collection was carried out in December 2002 and February 2003. Representative muscle, liver and fat samples were taken from each deer. Four aliquots from each sample were collected by carbon-fibre fractionation. All possible 209 CB congeners were measured with minimum isomeric interference. Fractions were analyzed using high resolution gas chromatography (HRGC) and high resolution mass spectrometry (HRMS). Concentrations of identified compounds were calculated using mean relative response factors determined from calibration standard runs. Results showed that 55 of 209 CB congeners were not detected in the fat samples. The mean level of CB homologues in the 2003 study was 39 ng/g for the lipid basis in the muscle, 92 ng/g in the liver, and 20.5 ng/g in the fat. Di-ortho CBs constituted 72 per cent to 74 per cent of CBs for the samples. Concentration levels of SCBs were lower than studies conducted in previous years. It was concluded that overall levels of SCBs and SCBs TEQ in the liver and muscle samples in 2003 significantly declined when compared with levels observed in 1997, 1999, and 2001. 6 refs., 4 tabs.

  13. Is there any Correlation between Magnetic Resonance Imaging Features of Breast Lesions of BIRADS Category 4 with Histopathologic Results?

    Directory of Open Access Journals (Sweden)

    Maryam Farghadani

    2017-01-01

    Full Text Available Background: To evaluate the correlation of magnetic resonance imaging (MRI features of breast lesions of Breast Imaging Reporting and Database System (BI-RADS category 4 with histopathologic results. Materials and Methods: In a prospective study between December 2013 and April 2015, patients with suspicious mammographic and/or ultrasound findings referred for Breast MRI were evaluated. Patients with lesions of BI-RADS category 4 were enrolled with a written informed consent. In each patient, mass lesion (ML or nonmass lesion (NML was determined, and different characteristics of the lesions were recorded. A follow-up program was taken with mean 3–12 months. Patients who underwent core needle biopsy or open biopsy were summoned. Results: Seventy-eight females aged 24–67 years (mean 43.1 ± 8.8 met the inclusion criteria and had adequate samples for histopathologic study. Twenty-nine (37.2% patients had ML and 49 (62.8% patients had NML. Tissue sampling in 63 (80.7% patients was through core needle biopsy and in 15 (19.2% patients through surgery. A wide spectrum of benign and malignant pathologic diagnoses was seen. In statistical analysis, none of the MRI features has a significant correlation with any specific histopathologic diagnosis (P = 0.185. However, the relation between the MRI category (ML or NML and pathology results was significant at level of 0.1 (P = 0.06. Conclusion: This study showed that a wide spectrum of histopathologic results is seen in BI-RADS category 4. However, in this sample volume, none of the MRI features in this BI-RADS category has a significant correlation with any specific histopathologic diagnosis.

  14. Is there any Correlation between Magnetic Resonance Imaging Features of Breast Lesions of BIRADS Category 4 with Histopathologic Results?

    Science.gov (United States)

    Farghadani, Maryam; Soofi, Ghazale Jamalipoor; Sarrami, Amir Hossein

    2017-01-01

    To evaluate the correlation of magnetic resonance imaging (MRI) features of breast lesions of Breast Imaging Reporting and Database System (BI-RADS) category 4 with histopathologic results. In a prospective study between December 2013 and April 2015, patients with suspicious mammographic and/or ultrasound findings referred for Breast MRI were evaluated. Patients with lesions of BI-RADS category 4 were enrolled with a written informed consent. In each patient, mass lesion (ML) or nonmass lesion (NML) was determined, and different characteristics of the lesions were recorded. A follow-up program was taken with mean 3-12 months. Patients who underwent core needle biopsy or open biopsy were summoned. Seventy-eight females aged 24-67 years (mean 43.1 ± 8.8) met the inclusion criteria and had adequate samples for histopathologic study. Twenty-nine (37.2%) patients had ML and 49 (62.8%) patients had NML. Tissue sampling in 63 (80.7%) patients was through core needle biopsy and in 15 (19.2%) patients through surgery. A wide spectrum of benign and malignant pathologic diagnoses was seen. In statistical analysis, none of the MRI features has a significant correlation with any specific histopathologic diagnosis (P = 0.185). However, the relation between the MRI category (ML or NML) and pathology results was significant at level of 0.1 (P = 0.06). This study showed that a wide spectrum of histopathologic results is seen in BI-RADS category 4. However, in this sample volume, none of the MRI features in this BI-RADS category has a significant correlation with any specific histopathologic diagnosis.

  15. [Histopathologic study of chronic sinusitis].

    Science.gov (United States)

    Wayoff, M; Parache, R M; Bodelet, B; Gazel, P

    1983-01-01

    The conventional histopathology of the sinus is a criterium for the therapeutic indication, since it is possible to distinguish between granulomatous chronic sinusitis, chronic sinusitis with oedema and nasal polyposis. Each one of these clinical pictures has his own etiology and requires a specific therapeutic approach.

  16. Histopathological spectrum of Psoriasiform dermatitis

    Directory of Open Access Journals (Sweden)

    PL Jayalakshmy

    2016-09-01

    Full Text Available Background: Psoriasiform dermatitis is a frequently encountered terminology in a wide variety of inflammatory dermatoses. It often poses challenges to both dermatologists and pathologists alike. Clinical features when considered alone may not be reliable, as they vary with both disease duration and treatment. On the contrary, histopathological material constitutes definite hard evidence, which can be preserved and will continue to be available for future review. The objective of the study is to study the histopathological findings in Psoriasiform dermatitis.Materials and Methods: This is a retrospective study in a tertiary care centre in Kerala, South India. All cases diagnosed as Psoriasis or mentioned as one of the differential diagnosis were included. The material included 66 skin biopsies. Patients with a previous diagnosis of the same were excluded from the study.Results: The lesions comprised 9% of the total number of skin biopsies. The age distribution pattern revealed that the highest percentage was in the 41-60 year age group (50% with a male preponderance of 65%. Psoriasis was the most common lesion (60.60% and Lichen simplex chronicus was the second most common lesion (10.60 %, encountered in the study.Conclusion: Some of the histopathological features are specific and characteristic for each entity like Psoriasis, Pityriasis rubra pilaris, Pityriasis rosea and Inflammatory linear verrucous epidermal nevus whereas some overlap in lesions like Prurigo nodularis, Lichen simplex chronicus and Allergic contact dermatitis. Hence, combination of proper clinical observation and histopathological study will give a conclusive diagnosis.

  17. Museums and disease: using tissue archive and museum samples to study pathogens.

    Science.gov (United States)

    Tsangaras, Kyriakos; Greenwood, Alex D

    2012-01-20

    Molecular studies of archival and fossil samples have traditionally focused on the nucleic acids derived from the host species. However, there has recently been an increase in ancient DNA research on the identification and characterization of infectious agents within the hosts. The study of pathogens from the past provides great opportunities for discovering the causes of historical infection events, characterizing host-microorganism co-evolution and directly investigating the evolution of specific pathogens. Several research teams have been able to isolate and characterize a variety of different bacterial, parasite and viral microorganisms. However, this emerging field is not without obstacles. The diagenetic processes that make ancient DNA research generally difficult are also impediments to ancient pathogen research and perhaps more so given that their DNA may represent an even rarer proportion of the remaining nucleic acids in a fossil sample than host DNA. However, studies performed under controlled conditions and following stringent ancient DNA protocols can and have yielded reliable and often surprising results. This article reviews the advantages, problems, and failures of ancient microbiological research.

  18. Advancements in mass spectrometry for biological samples: Protein chemical cross-linking and metabolite analysis of plant tissues

    Energy Technology Data Exchange (ETDEWEB)

    Klein, Adam [Iowa State Univ., Ames, IA (United States)

    2015-01-01

    This thesis presents work on advancements and applications of methodology for the analysis of biological samples using mass spectrometry. Included in this work are improvements to chemical cross-linking mass spectrometry (CXMS) for the study of protein structures and mass spectrometry imaging and quantitative analysis to study plant metabolites. Applications include using matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) to further explore metabolic heterogeneity in plant tissues and chemical interactions at the interface between plants and pests. Additional work was focused on developing liquid chromatography-mass spectrometry (LC-MS) methods to investigate metabolites associated with plant-pest interactions.

  19. Histopathological, immunohistochemical criteria and confocal laser-scanning data of arthrofibrosis.

    Science.gov (United States)

    Ruppert, M; Theiss, C; Knöß, P; Kendoff, D; Krukemeyer, M G; Schröder, N; Brand-Saberi, B; Gehrke, T; Krenn, V

    2013-11-01

    Arthrofibrosis (af) is defined as a fibrosing disease of the synovial membrane, after joint operations, with painful restricted range of motion. The aim of this paper was to describe the histopathological substrate of af, hitherto only defined by clinical criteria. Based on a group of 222 tissue samples, the characteristic changes to af were analyzed. The control group comprised 29 cases with neosynovialis of the indifferent type. Due to cytoplasmic SM-actin positivity and the absence of specific cytoplasmic reactivity in CD 68 representation, af fibroblasts were characterized as myofibroblasts. In confocal laser-scanning microscopy, β-catenin-positive aggregates were detected in the cytoplasm. Over and above this, unequivocal colocalization of β-catenin and the tight junction protein ZO-1 became manifest, particularly on the cell membrane and, partly, in the cytoplasm. A threshold value of 20 β-catenin-positive cells/HPF was determined. This enables the histopathological diagnosis of an af to be made (sensitivity: 0.733, specificity: 0.867). Af is a fibrosing disease of the synovial membrane with variable grade of fibrotization (fibroblast cellularity). A threshold value of 20 β-catenin-positive fibroblasts per HPF was defined, which enables the histopathological diagnosis of af. Copyright © 2013 Elsevier GmbH. All rights reserved.

  20. 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.

  1. Application of tissue microarray for atherectomized tissues from peripheral arterial disease.

    Science.gov (United States)

    Kim, Se Hoon; Kim, Dong-Min; Shim, Hyosup; Choi, Junjeong; Park, Seong Hwan; Song, Seung Min; Shin, Young Ho; Choi, Donghoon

    2011-09-15

    It is not easy to apply tissue microarray (TMA) to atherectomized tissues from peripheral arterial disease because of their physical properties. We introduce a new TMA application technique for atherectomized tissues. Using a pre-made plastic TMA cassette and TMA punch device, we successfully made the TMA block containing 40 vertically oriented atherectomized tissue samples from 10 patients. The histogram of surface areas of tissue cores in the TMA showed a bell-shaped distribution, whereas that of conventionally embedded tissues showed wide distribution. This finding suggests that the TMA method might be a better way of vertical embedding than the conventional method. A TMA block prepared by our method enabled a simultaneous evaluation of the histopathology of vertically oriented atherectomized tissues and the correlation between them with intravascular ultrasound image. In addition, this new method might be applied to various tissues in different ways. Copyright © 2011 Elsevier GmbH. All rights reserved.

  2. Metastatic pheochromocytoma: clinical, genetic, and histopathologic characteristics

    Science.gov (United States)

    Zelinka, Tomáš; Musil, Zdeněk; Dušková, Jaroslava; Burton, Deborah; Merino, Maria J; Milosevic, Dragana; Widimský, Jiří; Pacak, Karel

    2011-01-01

    Background Pheochromocytomas are tumors arising from chromaffin tissue located in the adrenal medulla associated with typical symptoms and signs which may occasionally develop metastases, which are defined as the presence of tumor cells at sites where these cells are not found. This retrospective analysis was focused on clinical, genetic, and histopathologic characteristics of primary metastatic versus primary benign pheochromocytomas. Materials and methods We identified 41 subjects with metastatic pheochromocytoma and 108 subjects with apparently benign pheochromocytoma. We assessed dimension and biochemical profile of the primary tumor, age at presentation, and time to develop metastases. Results Subjects with metastatic pheochromocytoma presented at a significantly younger age (41.4±14.7 vs. 50.2±13.7 years; Ppheochromocytomas. No significant differences were found in the incidence of genetic mutations in both groups of subjects (25.7 % in the metastatic group and 14.7 % in the benign group; P=0.13). From available histopathologic markers of potential malignancy, only necrosis occurred more frequently in subjects with metastatic pheochromocytoma (27.6 % vs. 0 %; Ppheochromocytoma and age of its first presentation are two independent risk factors associated with the development of metastatic disease. PMID:21692797

  3. Static Bone Cavity of the Mandible: Computed Tomography Findings with Histopathologic Correlation

    Energy Technology Data Exchange (ETDEWEB)

    Minowa, K.; Inoue, N.; Izumiyama, Y.; Ashikaga, Y.; Chu, B.; Maravilla, K.R.; Totsuka, Y.; Nakamura, M. [Hokkaido Univ., Sapporo (Japan). Depts. of Radiology, Oral Health Science, and Oral Patho-biological Science

    2006-09-15

    Purpose: To review computed tomography (CT) findings of histopathologically examined static bone cavities in order to determine whether an additional pathogenesis may play a role in this disease. Material and Methods: Four patients with histopathologically examined static bone cavities were included in this retrospective study. Location, appearance of bone remodeling, tissue characteristics, and contrast enhancement of the cavity were assessed on CT images. CT findings were then compared with the histopathological findings. Results: Static bone cavity was found in the lingual molar region in three patients and in the lingual cuspid region of the mandible in one patient. Both fatty and soft tissues were present in the cavities of all four patients. Attenuation of the soft tissue in the cavities was found to be different from that of the submandibular gland. The soft tissue showed enhancement with contrast-enhanced CT in three patients. For all patients, the histopathologic content of the static bone cavity included fat, soft tissue, and abnormal vasculature. The thickened vein wall in the abnormal vasculature was observed. Aberrant tissue of the submandibular gland was not found in any of the static bone cavities. Conclusion: Contrast enhancement of the soft tissue on the contrast-enhanced CT images suggests the presence of vasculature in the cavities. Histopathological examination confirmed the presence of fatty tissue and dilated abnormal vessels, and the absence of salivary gland tissue in the cavities. These findings show that vascular structures are prominent in tissues found in static bone cavities.

  4. A comprehensive factorial design study of variables affecting protein extraction from formalin-fixed kidney tissue samples.

    Science.gov (United States)

    Araújo, J E; Oliveira, E; Otero-Glez, A; Santos Nores, J; Igrejas, G; Lodeiro, C; Capelo, J L; Santos, H M

    2014-02-01

    Formalin-fixed tissues are an important source of biological samples for biomedical research. However, proteomics analysis of formalin-fixed tissues has been set aside by formalin-induced protein modifications, which reduce protein extraction efficiency. In this study, a two level full factorial experimental design (2(4)) was used to determine the effects of the extracting conditions in the efficiency of protein recovery from formalin-fixed kidney samples. The following variables were assessed: temperature of extraction, pH of extraction, composition of the extracting buffer and the use ultrasonic energy applied with probe. It is clearly demonstrated that when hating and ultrasonic energy are used in conjunction, a 7-fold increase (p protein extraction is obtained if compared to extracting conditions for which neither heating nor ultrasonic energy are used. The optimization study was done following the amount of protein extracted by UV (Nanodrop(®) technology, protein ABS at 280 nm) and by 1D SDS-PAGE. Extracts obtained with the optimized conditions were subjected to LC-MALDI MS/MS. A total of 112 proteins were identified.

  5. Analysis of azole fungicides in fish muscle tissues: Multi-factor optimization and application to environmental samples.

    Science.gov (United States)

    Zhong, Yuanhong; Chen, Zhi-Feng; Liu, Shuang-Shuang; Dai, Xiaoxin; Zhu, Xinping; Zheng, Guangming; Liu, Shugui; Liu, Guoguang; Cai, Zongwei

    2017-02-15

    Azole fungicides have been reported to be accumulated in fish tissue. In this study, a sensitive and robust method using high-performance liquid chromatography-tandem mass spectrometry combined with ultrasonic extraction, solid-liquid clean-up, liquid-liquid extraction and solid-phase extraction (SPE) for enrichment and purification have been proposed for determination of azole fungicides in fish muscle samples. According to the results of non-statistical analysis and statistical analysis, ethyl acetate, primary secondary amine (PSA) and mixed-mode cation exchange cartridge (MCX) were confirmed as the best extraction solvent, clean-up sorbent and SPE cartridge, respectively. The satisfied recoveries (81.7-104%) and matrix effects (-6.34-7.16%), both corrected by internal standards, were performed in various species of fish muscle matrices. Method quantification limits of all azoles were in the range of 0.07-2.83ng/g. This optimized method was successfully applied for determination of the target analytes in muscle samples of field fish from Beijiang River and its tributaries. Three azole fungicides including climbazole, clotrimazole and carbendazim were detected at ppb levels in fish muscle tissues. Therefore, this analytical method is practical and suitable for further clarifying the contamination profiles of azole fungicides in wild fish species. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Graph run-length matrices for histopathological image segmentation.

    Science.gov (United States)

    Tosun, Akif Burak; Gunduz-Demir, Cigdem

    2011-03-01

    The histopathological examination of tissue specimens is essential for cancer diagnosis and grading. However, this examination is subject to a considerable amount of observer variability as it mainly relies on visual interpretation of pathologists. To alleviate this problem, it is very important to develop computational quantitative tools, for which image segmentation constitutes the core step. In this paper, we introduce an effective and robust algorithm for the segmentation of histopathological tissue images. This algorithm incorporates the background knowledge of the tissue organization into segmentation. For this purpose, it quantifies spatial relations of cytological tissue components by constructing a graph and uses this graph to define new texture features for image segmentation. This new texture definition makes use of the idea of gray-level run-length matrices. However, it considers the runs of cytological components on a graph to form a matrix, instead of considering the runs of pixel intensities. Working with colon tissue images, our experiments demonstrate that the texture features extracted from "graph run-length matrices" lead to high segmentation accuracies, also providing a reasonable number of segmented regions. Compared with four other segmentation algorithms, the results show that the proposed algorithm is more effective in histopathological image segmentation.

  7. Interferon-λ4 (IFNL4 transcript expression in human liver tissue samples.

    Directory of Open Access Journals (Sweden)

    Ahmad Amanzada

    Full Text Available Eradication of hepatitis C virus (HCV infection, both spontaneous and treatment-induced, is marked by the wildtype allele C of a single nucleotide polymorphism upstream of the IL28B gene, rs12979860. This favorable allele was recently described to be in linkage disequilibrium with the wildtype allele TT of a dinucleotide polymorphism, ss469415590, located within a new protein-coding gene. While the TT allele introduces a frame-shift and disrupts the open reading frame, only the variant allele, ΔG, creates a novel type III interferon (IFN protein, IFN-λ4/IFNL4. Absence of IFNL4 is thus supposed to favor resolution of HCV infection. As to date IFNL4 mRNA transcription has only been investigated in polyI:C-stimulated primary human hepatocytes and not yet in HCV infection in vivo, this study analyzed IFNL4 mRNA expression in human liver biopsy specimens. Samples were obtained from patients with a broad panel of disorders including no liver disease, liver diseases of non-viral etiology, chronic hepatitis B and chronic hepatitis C. Hepatic IFNL4 transcripts were detectable exclusively in a subgroup of chronic hepatitis C patients (24/45. Their amounts were positively related to liver HCV RNA copy numbers (p = 0.0023, r = 0.56 suggesting that the hepatic viral load influences IFNL4 transcription irrespective of IFNL4 governing genotype. Both, the IFNL4 creating allele ΔG (p<0.0001 and actual IFNL4 transcription (p = 0.0015 were found to be correlated to the activation of IFN stimulatory genes (ISGs. By contrast, IFNL4 ss469415590 genotypes were not found to be related to IFN-λ2/3/IL28 or IFN-λ1/IL29 gene expression. In conclusion, this study is the first report on intrahepatic transcript levels of the recently discovered IFNL4 gene. Data indicate that HCV infection in particular might activate IFNL4 transcription in the liver. It provides a possible explanation as to why hepatitis C patients show ISG stimulation in their livers in the

  8. Cascaded discrimination of normal, abnormal, and confounder classes in histopathology: Gleason grading of prostate cancer

    Directory of Open Access Journals (Sweden)

    Doyle Scott

    2012-10-01

    Full Text Available Abstract Background Automated classification of histopathology involves identification of multiple classes, including benign, cancerous, and confounder categories. The confounder tissue classes can often mimic and share attributes with both the diseased and normal tissue classes, and can be particularly difficult to identify, both manually and by automated classifiers. In the case of prostate cancer, they may be several confounding tissue types present in a biopsy sample, posing as major sources of diagnostic error for pathologists. Two common multi-class approaches are one-shot classification (OSC, where all classes are identified simultaneously, and one-versus-all (OVA, where a “target” class is distinguished from all “non-target” classes. OSC is typically unable to handle discrimination of classes of varying similarity (e.g. with images of prostate atrophy and high grade cancer, while OVA forces several heterogeneous classes into a single “non-target” class. In this work, we present a cascaded (CAS approach to classifying prostate biopsy tissue samples, where images from different classes are grouped to maximize intra-group homogeneity while maximizing inter-group heterogeneity. Results We apply the CAS approach to categorize 2000 tissue samples taken from 214 patient studies into seven classes: epithelium, stroma, atrophy, prostatic intraepithelial neoplasia (PIN, and prostate cancer Gleason grades 3, 4, and 5. A series of increasingly granular binary classifiers are used to split the different tissue classes until the images have been categorized into a single unique class. Our automatically-extracted image feature set includes architectural features based on location of the nuclei within the tissue sample as well as texture features extracted on a per-pixel level. The CAS strategy yields a positive predictive value (PPV of 0.86 in classifying the 2000 tissue images into one of 7 classes, compared with the OVA (0.77 PPV and OSC

  9. The problem with medical research on tissue and organ samples taken in connection with forensic autopsies in France.

    Science.gov (United States)

    Rougé-Maillart, C; Dupont, V; Jousset, N

    2016-02-01

    Currently, in France, it is legally impossible to conduct scientific research on tissue and organ samples taken from forensic autopsies. In fact, the law schedules the destruction of such samples at the end of the judicial investigation, and the common law rules governing cadaver research cannot be applied to the forensic context. However, nothing seems in itself to stand in the way of such research since, despite their specific nature, these samples from forensic autopsies could be subject, following legislative amendments, to common law relating to medical research on samples taken from deceased persons. But an essential legislative amendment, firstly to allow the Biomedicine Agency to become authorized to issue a research permit and secondly, to change the research conditions in terms of the non-opposition of the deceased to said research. Such an amendment would be a true breakthrough because it would allow teams to continue to move forward calmly in research, and allow this research to be placed within a legal framework, which would promote international exchanges.

  10. Masses in the Neck: A Histopathological Analysis of 213 Cases In ...

    African Journals Online (AJOL)

    Masses in the Neck: A Histopathological Analysis of 213 Cases In South ... of Calabar Teaching Hospital, Calabar, Nigeria are caused by lymphoid tissues. ... The reason for this unusual presentation in an area of high prevalence rate of ...

  11. Connective Tissue Naevus

    Directory of Open Access Journals (Sweden)

    Bhat Ramesh M

    1999-01-01

    Full Text Available A young adult female patient of connective tissue naevus presented with papules and indurated plaques on both les and left arm. Histopathology showed increased amount of collagen in the dermis. Osteopoikilosis was absent.

  12. Salivary Duct Cyst: Histo-pathologic Correlation

    Directory of Open Access Journals (Sweden)

    Divya Vinayachandran

    2013-01-01

    Full Text Available Non-neoplastic cysts of the salivary glands are uncommon and represent 2-5% of all salivary gland lesions. They are mainly mucoceles or salivary duct cysts. Unlike a mucocele, which is surrounded by granulation tissue, the salivary duct cyst is lined by epithelium. Salivary duct cysts are more common in the oral minor salivary glands and rarely occur in the major salivary glands, show a marked predilection for the superficial lobe of the parotid, and represent 10% of all salivary gland cysts. Neoplastic differentiation of the lining of the salivary duct cyst has been reported. We report a case of a salivary duct cyst of the left parotid gland, with a review of radiographic and histopathologic features.

  13. Histopathology of marine vibrio wound infections.

    Science.gov (United States)

    Beckman, E N; Leonard, G L; Castillo, L E; Genre, C F; Pankey, G A

    1981-12-01

    Although marine vibrio wound infections and septicemia are being reported with increasing frequency, description of the histopathologic changes has been scanty. The histologic alterations in three patients with primary marine vibrio wound infections are presented. The lesions are characterized by intense acute cellulitis of the subcutis with much tissue destruction and extension into the adjacent dermis. The superficial dermis is devitalized and lacks an inflammatory cellular infiltrate. Subepidermal noninflammatory bullae are formed. Many organisms are seen both within the areas of intense acute inflammation and in devitalized areas. Organisms and inflammation are especially oriented around vessels, with associated acute vasculitis. It is concluded that the morphologic picture in marine vibrio wound infections is nonspecific yet characteristic.

  14. Histopathologic effects of formaldehyde exposure on rat kidney

    Directory of Open Access Journals (Sweden)

    M.J. Golalipour

    2007-01-01

    Full Text Available AbstractBackground and purpose: Formaldehyde is a chemical traditionally used for fixing the cadaver. It is vaporized during dissection and practical studying on cadaver. Studies show that this vapour can cause some clinical sympotms such as throat, eye, skin and nasal irritation.This study was designed to determine the histopathological changes of rat kidney tissue exposed to formaldehyde for 18 weeks.Materials and Methods: This study was performed on 28, 6-7 weeks postnatal albino Wistar rats. The rats were divided into 3 case groups (E1: 4hrs/d, 4d/w; E2: 2hrs/d, 4d/w; E3: 2hrs/d, 2d/w and one control group (C. The kidney specimens were sectioned and stained with H&E technique for histopathological study.Results: In all histopathology sections of groups E1, E2 and E3, the following similar changes were observed: Mild congestion in the glumeroles, focal congestion and vacuolar (hydropic degeneration of tubular cells only mild non-specific congestion in renal vessels. There were no evidences of fibrotic change or inflammatory cells infiltration among interstitial tissue. Also there were no abnormalities in the staining of nucleus and cytoplasm. In Control group (C, no histopathologic changes were observed.Conclusion: The results of this study showed that formaldehyde vapour with a concentrations used in our study, can not induce histopathologic changes which could be detectable by light microscope. Also, there is no direct relationship between the duration of exposure to formaldehyde vapour and the intensity of histopathologic changes in the kidney.

  15. Pityriasis rosea: A histopathologic study

    Directory of Open Access Journals (Sweden)

    Prasad D

    2000-01-01

    Full Text Available The histopathologic features of pityriasis rosea (PR have been considered by many authors to be compatible with those of dermatitis, but it is not surprising that histological difference may exist between PR and dermatitis. fifty patients of pityriasis rosea were included in this study. We studied the 50 biopsies including 9 biopsies from herald patches. The characteristic features were; absence or decrease of the granular cell layer, extravasation of red blood cells in papillary dermis and partly into the epidermis, dyskeratosis, liquefaction of basal cells, homogenization of papillary collagen, intraepidermal vesicles in apparently dry skin. In addition of these features, we observed prominent delling of epidermis and a typical spongiotic pattern not reported earlier. The biopsies from herald patches showed minor differences from the secondary lesions. We also compared the histopathological features of recent lesions with older lesions.

  16. Biochemical, histopathological and ultra structural profile after pulsed water medication of enrofloxacin in broiler chickens

    Directory of Open Access Journals (Sweden)

    Veerapandian Sureshkumar

    2013-07-01

    Full Text Available Aim: A pharmacological study was undertaken to evaluate the safety and adverse effects of enrofloxacin administration in broiler chickens by assessing the serum biochemical parameters, associated histopathological and ultra structural changes in liver and kidney.Materials and Methods: Birds in the treatment group were administered with enrofloxacin at the recommended therapeutic dose 10mg/kg body weight via drinking water for five successive days, while the control group (untreated group received non medicated water. Serum biochemical parameters viz., total protein, albumin, lactate dehydrogenase, alkaline phosphatase, creatine kinase, lipase, triglyceride, gamma glutamyl transferase, urea, uric acid and creatinine were estimated at 24hour and 48hour intervals during the dosing and withdrawal periods, respectively. Liver and kidney tissue samples collected from 1, 3, 5, 7, and 9 days post treatment groups were subjected to histopathological and ultrastructural examinations.Results: There was no significant change (p>0.05 in total protein, albumin, lactate dehydrogenase, alkaline phosphatase, creatine kinase, lipase, triglyceride and urea levels in the enrofloxacin administered broiler chickens at all the time points evaluated. However, a significant increase (p<0.05 in gamma glutamyl transferase, uric acid and creatinine levels were th observed after the 4 dose of the enrofloxacin and on day 1 post treatment. During the withdrawal period, the elevated levels declined gradually and showed the trend towards control values as evidenced by a statistically insignificant difference on 3, 5, 7 and 9 days post treatment when compared to that of control group. These biochemical changes were substantiated by histopathological and ultrastructural changes elicited in liver and kidney.Conclusion: The reversible trend observed in serum biochemical parameters, histopathological and ultra structural alterations in liver and kidney during the withdrawal period

  17. A New Sample Substrate for Imaging and Correlating Organic and Trace Metal Composition in Biological Cells and Tissues

    Energy Technology Data Exchange (ETDEWEB)

    Miller,L.; Wang, Q.; Smith, R.; Zhong, H.; Elliott, D.; Warren, J.

    2007-01-01

    Many disease processes involve alterations in the chemical makeup of tissue. Synchrotron-based infrared (IR) and X-ray fluorescence (XRF) microscopes are becoming increasingly popular tools for imaging the organic and trace metal compositions of biological materials, respectively, without the need for extrinsic labels or stains. Fourier transform infrared microspectroscopy (FTIRM) provides chemical information on the organic components of a material at a diffraction-limited spatial resolution of 2-10 {mu}m in the mid-infrared region. The synchrotron X-ray fluorescence (SXRF) microprobe is a complementary technique used to probe trace element content in the same systems with a similar spatial resolution. However to be most beneficial, it is important to combine the results from both imaging techniques on a single sample, which requires precise overlap of the IR and X-ray images. In this work, we have developed a sample substrate containing a gold grid pattern on its surface, which can be imaged with both the IR and X-ray microscopes. The substrate consists of a low trace element glass slide that has a gold grid patterned on its surface, where the major and minor parts of the grid contain 25 and 12 nm gold, respectively. This grid pattern can be imaged with the IR microscope because the reflectivity of gold differs as a function of thickness. The pattern can also be imaged with the SXRF microprobe because the Au fluorescence intensity changes with gold thickness. The tissue sample is placed on top of the patterned substrate. The grid pattern's IR reflectivity image and the gold SXRF image are used as fiducial markers for spatially overlapping the IR and SXRF images from the tissue. Results show that IR and X-ray images can be correlated precisely, with a spatial resolution of less than one pixel (i.e., 2-3 microns). The development of this new tool will be presented along with applications to paraffin-embedded metalloprotein crystals, Alzheimer's disease

  18. 42 CFR 493.1219 - Condition: Histopathology.

    Science.gov (United States)

    2010-10-01

    ... 42 Public Health 5 2010-10-01 2010-10-01 false Condition: Histopathology. 493.1219 Section 493....1219 Condition: Histopathology. If the laboratory provides services in the subspecialty of Histopathology, the laboratory must meet the requirements specified in §§ 493.1230 through 493.1256, §...

  19. Histopathology and the detection of avian bornavirus in the nervous system of birds diagnosed with proventricular dilatation disease.

    Science.gov (United States)

    Ouyang, N; Storts, R; Tian, Y; Wigle, W; Villanueva, I; Mirhosseini, N; Payne, S; Gray, P; Tizard, I

    2009-10-01

    Avian bornavirus (ABV) is currently considered a probable etiologic agent of proventricular dilatation disease (PDD) of psittacines. We tested 24 stored avian brain samples, processed for histopathology and retained following their submission for necropsy or histopathology to the Schubot Exotic Bird Center diagnostic laboratory in 1992. Thirteen of these samples were from birds diagnosed at that time as suffering from PDD. The remaining 11 samples were diagnosed as suffering from diseases other than PDD. Immunohistochemistry was performed using an antiserum directed against the ABV nucleoprotein (N-protein). Stained slides were read by an investigator unaware of their prior histopathology results. Cells containing ABV N-protein were present in the nervous tissues of all 13 PDD cases. One bird not previously diagnosed with PDD also had ABV N-protein in its brain. A review of this bird's necropsy report indicated that it was, most probably, also suffering from PDD. The remaining 10 non-PDD birds had no detectable N-protein in their brains. The N-protein was present in the cerebrum, cerebellum and spinal cord. These findings support other studies that indicate that ABV is an etiological agent of PDD.

  20. Blind Biobanking of the Prostatectomy Specimen: Critical Evaluation of the Existing Techniques and Development of the New 4-Level Tissue Extraction Model With High Sampling Efficacy.

    Science.gov (United States)

    Tolkach, Yuri; Eminaga, Okyaz; Wötzel, Fabian; Huss, Sebastian; Bettendorf, Olaf; Eltze, Elke; Abbas, Mahmoud; Imkamp, Florian; Semjonow, Axel

    2017-03-01

    Fresh tissue is mandatory to perform high-quality translation studies. Several models for tissue extraction from prostatectomy specimens without guidance by frozen sections are already introduced. However, little is known about the sampling efficacy of these models, which should provide representative tissue in adequate volumes, account for multifocality and heterogeneity of tumor, not violate the routine final pathological examination, and perform quickly without frozen section-based histological control. The aim of the study was to evaluate the sampling efficacy of the existing tissue extraction models without guidance by frozen sections ("blind") and to develop an optimized model for tissue extraction. Five hundred thirty-three electronic maps of the tumor distribution in prostates from a single-center cohort of the patients subjected to radical prostatectomy were used for analysis. Six available models were evaluated in silico for their sampling efficacy. Additionally, a novel model achieving the best sampling efficacy was developed. The available models showed high efficacies for sampling "any part" from the tumor (up to 100%), but were uniformly low in efficacy to sample all tumor foci from the specimens (with the best technique sampling only 51.6% of the all tumor foci). The novel 4-level extraction model achieved a sampling efficacy of 93.1% for all tumor foci. The existing "blind" tissue extraction models from prostatectomy specimens without frozen sections control are suitable to target tumor tissues but these tissues do not represent the whole tumor. The novel 4-level model provides the highest sampling efficacy and a promising potential for integration into routine. Prostate 77: 396-405, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  1. Sample Preparation and Staining Methods for Two-Dimensional Polyacrylamide Gel Electrophoresis of Proteins from Animal Tissues

    Directory of Open Access Journals (Sweden)

    Levente Czegledi

    2010-05-01

    Full Text Available Proteomics in animal science as well as in other biological sciences is a significant tool in the post-genomic era. In proteomic studies the presence and relative abundance of expressed proteins of a cell, tissue or biological fluid is studied. Recently, the whole genome of more and more domestic animal species is known, but genes and the transcribed mRNA have no direct effect on biological systems as they are regulated by proteins, which explain the importance of proteomics. The most common tool in proteomic approach is the two-dimensional polyacrylamide gel electrophoresis (2D PAGE, when proteins are separated by their isoelectric point followed by their mass separation as a second dimension. In this study authors used different sample preparation and protein staining methods on meat,  liver and blood plasma and carried out 2D PAGE experiments. The most appropriate sample preparation methods are described in this paper. We concluded that depletion of major proteins in plasma is required but not necessary for meat and liver samples.

  2. The histopathology of Candida albicans invasion in neonatal rat tissues and in the human blood-brain barrier in culture revealed by light, scanning, transmission and immunoelectron microscopy scanning

    OpenAIRE

    Lossinsky, A.S.; de Jong, A.; Fiala, M; Mukhtar, M; Buttle, K.F.; Ingram, M.

    2006-01-01

    The present studies examined the effects of Candida albicans yeast and hyphal morphologies on tissue pathologies and transmigration properties of the fungus in two experimental models: 1) an in vivo, neonatal rat model, and 2) a cell culture model of human brain microvascular endothelial cells (ECs) (BMVEC). We inoculated a hyphae-producing strain (CAI4-URA3) and a non-hyphae-producing strain (CAI4) of C. albicans into 4-10 day old rats and BMVEC cultures. ...

  3. Virus isolation vs RT-PCR: which method is more successful in detecting VHSV and IHNV in fish tissue sampled under field conditions?

    DEFF Research Database (Denmark)

    Knüsel, R.; Bergmann, S. M.; Einer-Jensen, Katja;

    2007-01-01

    (total of 859 fish) originating from a field survey on the occurrence of VHSV and IHNV in farmed and wild salmonids in Switzerland. These samples represented all sites with fish that were either identified as virus-positive by means of virus isolation (three sites, four positive tissue sample pools) and......This study compared the results of reverse transcription-polymerase chain reaction (RT-PCR) and traditional virus isolation on cell culture in detection of viral haemorrhagic septicaemia virus (VHSV) and infectious haematopoietic necrosis virus (IHNV). RT-PCR was used for 172 tissue sample pools....../or demonstrated positive anti-VHSV-antibody titres (83 sites, 121 positive blood samples) in a serum plaque neutralization test (SPNT). The RT-PCR technique confirmed the four VHSV-positive tissue sample pools detected by virus isolation and additionally identified one VHSV-positive sample that showed positive...

  4. Contaminants in white-tailed deer tissue from the Great Swamp National Wildlife Refuge Morris and Somerset Counties, New Jersey: Results of 1988 sampling and analysis

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — White-tailed deer (Odocoileus virginianus) tissues were sampled during the December, 1988, public deer hunt at the Great Swamp National Wildlife Refuge (GSNWR) to...

  5. ALP gene expression in cDNA samples from bone tissue engineering using a HA/TCP/Chitosan scaffold

    Science.gov (United States)

    Stephanie, N.; Katarina, H.; Amir, L. R.; Gunawan, H. A.

    2017-08-01

    This study examined the potential use of hydroxyapatite (HA)/tricalcium phosphate (TCP)/Chitosan as a bone tissue engineering scaffold. The potential for using HA/TCP/chitosan as a scaffold was analyzed by measuring expression of the ALP osteogenic gene in cDNA from bone biopsies from four Macaque nemestrina. Experimental conditions included control (untreated), treatment with HA/TCP 70:30, HA/TCP 50:50, and HA/TCP/chitosan. cDNA samples were measured quantitively with Real-Time PCR (qPCR) and semi-quantitively by gel electrophoresis. There were no significant differences in ALP gene expression between treatment subjects after two weeks, but the HA/TCP/chitosan treatment gave the highest level of expression after four weeks. The scaffold using the HA/TCP/chitosan combination induced a higher level of expression of the osteogenic gene ALP than did scaffold without chitosan.

  6. Routine sample preparation and HPLC analysis for ascorbic acid (vitamin C) determination in wheat plants and Arabidopsis leaf tissues.

    Science.gov (United States)

    Szalai, Gabriella; Janda, T; Pál, Magda

    2014-06-01

    Plants have developed various mechanisms to protect themselves against oxidative stress. One of the most important non-enzymatic antioxidants is ascorbic acid. There is thus a need for a rapid, sensitive method for the analysis of the reduced and oxidised forms of ascorbic acid in crop plants. In this paper a simple, economic, selective, precise and stable HPLC method is presented for the detection of ascorbate in plant tissue. The sensitivity, the short retention time and the simple isocratic elution mean that the method is suitable for the routine quantification of ascorbate in a high daily sample number. The method has been found to be better than previously reported methods, because of the use of an economical, readily available mobile phase, UV detection and the lack of complicated extraction procedures. The method has been tested on Arabidopsis plants with different ascorbate levels and on wheat plants during Cd stress.

  7. Validation of the LacTek test applied to spiked extracts of tissue samples: determination of performance characteristics.

    Science.gov (United States)

    Mitchell, J M; Yee, A J; McNab, W B; Griffiths, M W; McEwen, S A

    1999-01-01

    LacTek tests are competitive enzyme-linked immunosorbent assays intended for rapid detection of antimicrobial residues in bovine milk. In this study, the LacTek test protocol was modified for use with extracts of bovine tissue to detect beta-lactam, tetracycline, and sulfamethazine residues. Test performance characteristics--precision, accuracy, ruggedness, practicability, and analytical specificity and sensitivity--were investigated. Results suggest that LacTek tests can be easily adapted to detect antimicrobial residues in extracts of lean ground beef. However, positive samples may not contain residues at violative concentrations (i.e., Canadian maximum residue limits), and therefore, additional analysis would be required for final confirmation and quantitation (e.g., chromatography).

  8. Fine-needle aspiration and core needle biopsy: An update on 2 common minimally invasive tissue sampling modalities.

    Science.gov (United States)

    VanderLaan, Paul A

    2016-12-01

    Fine-needle aspiration (FNA) and core needle biopsy (CNB) represent 2 of the most common minimally invasive tissue sampling modalities. Although similar in many ways, there are significant differences in the collection, processing, interpretation, and suitability for ancillary testing that exist between FNA and CNB. This review provides a brief overview of the strengths and weaknesses of FNA compared with CNB, as well as an update regarding the landscape of recently published studies that investigate the organ-specific comparative performance metrics of FNA and CNB. A current understanding of the benefits and limitations of FNA and CNB will help the cytopathologist and the clinician alike to select the right procedure for the right patient at the right time. Cancer Cytopathol 2016;124:862-870. © 2016 American Cancer Society. © 2016 American Cancer Society.

  9. Parasitic worms and their histopathological effects in four sturgeon species from the southwest shores of the Caspian Sea

    Directory of Open Access Journals (Sweden)

    Mohammad Reza Noei

    2016-01-01

    Full Text Available This study conducted to provide the status of parasite communities of four sturgeon species viz. Acipenser stellatus, A. gueldenstaedtii, A. nudiventris, and Huso huso in the southwest of the Caspian Sea and their histopathological effects on the examined fishes. For this purpose a total of 93 individuals of four sturgeon species were caught in two fisheries regions from the southwest of the Caspian Sea (Guilan Province, Iran from March 2010 to May 2011. The histological slides of the infested tissues of the examined fishes were prepared for study of the histopathological effects of the parasites. Classical epidemiological variables, including mean intensity, prevalence, abundance and dominance were calculated for overall samples, grouped by season, geographical region, and sex. Five worm species, including two nematodes (Cucullanus   sphaerocephalus and Eustrongylides excisus larvae, one cestode (Bothrimonus fallax, one acanthocephalans (Leptorhynchoide plagicephalus and one digenean trematode (Skrjabinopsolus   semiarmatus were found in examined sturgeons and their histopathological effects on the fish tissues were assessed. Based on the results, the diversity of the parasites (including freshwater ones in the southern part of the Caspian Sea have decreased since the time of the first study in 1972. This may be related to unfavorable conditions in freshwater ecosystems.

  10. Reduction of sample size requirements by bilateral versus unilateral research designs in animal models for cartilage tissue engineering.

    Science.gov (United States)

    Orth, Patrick; Zurakowski, David; Alini, Mauro; Cucchiarini, Magali; Madry, Henning

    2013-11-01

    Advanced tissue engineering approaches for articular cartilage repair in the knee joint rely on translational animal models. In these investigations, cartilage defects may be established either in one joint (unilateral design) or in both joints of the same animal (bilateral design). We hypothesized that a lower intraindividual variability following the bilateral strategy would reduce the number of required joints. Standardized osteochondral defects were created in the trochlear groove of 18 rabbits. In 12 animals, defects were produced unilaterally (unilateral design; n=12 defects), while defects were created bilaterally in 6 animals (bilateral design; n=12 defects). After 3 weeks, osteochondral repair was evaluated histologically applying an established grading system. Based on intra- and interindividual variabilities, required sample sizes for the detection of discrete differences in the histological score were determined for both study designs (α=0.05, β=0.20). Coefficients of variation (%CV) of the total histological score values were 1.9-fold increased following the unilateral design when compared with the bilateral approach (26 versus 14%CV). The resulting numbers of joints needed to treat were always higher for the unilateral design, resulting in an up to 3.9-fold increase in the required number of experimental animals. This effect was most pronounced for the detection of small-effect sizes and estimating large standard deviations. The data underline the possible benefit of bilateral study designs for the decrease of sample size requirements for certain investigations in articular cartilage research. These findings might also be transferred to other scoring systems, defect types, or translational animal models in the field of cartilage tissue engineering.

  11. Radiation-Induced Liver Damage: Correlation of Histopathology with Hepatobiliary Magnetic Resonance Imaging, a Feasibility Study

    Energy Technology Data Exchange (ETDEWEB)

    Seidensticker, Max, E-mail: max.seidensticker@med.ovgu.de [Universitätsklinik Magdeburg, Klinik für Radiologie und Nuklearmedizin (Germany); Burak, Miroslaw [Pomeranian Medical University, Department of Diagnostic Imaging and Interventional Radiology (Poland); Kalinski, Thomas [Universitätsklinik Magdeburg, Institut für Pathologie (Germany); Garlipp, Benjamin [Universitätsklinik Magdeburg, Klinik für Allgemein-, Viszeral- und Gefäßchirurgie (Germany); Koelble, Konrad [Philipps Universität Marburg, Fachbereich Medizin der, Abteilung für Neuropathologie (Germany); Wust, Peter [Charité Universitätsmedizin Berlin, Klinik für Radioonkologie und Strahlentherapie (Germany); Antweiler, Kai [Universitätsklinik Magdeburg, Institut für Biometrie und Medizinische Informatik (Germany); Seidensticker, Ricarda; Mohnike, Konrad; Pech, Maciej; Ricke, Jens [Universitätsklinik Magdeburg, Klinik für Radiologie und Nuklearmedizin (Germany)

    2015-02-15

    PurposeRadiotherapy of liver malignancies shows promising results (radioembolization, stereotactic irradiation, interstitial brachytherapy). Regardless of the route of application, a certain amount of nontumorous liver parenchyma will be collaterally damaged by radiation. The functional reserve may be significantly reduced with an impact on further treatment planning. Monitoring of radiation-induced liver damage by imaging is neither established nor validated. We performed an analysis to correlate the histopathological presence of radiation-induced liver damage with functional magnetic resonance imaging (MRI) utilizing hepatobiliary contrast media (Gd-BOPTA).MethodsPatients undergoing local high-dose-rate brachytherapy for whom a follow-up hepatobiliary MRI within 120 days after radiotherapy as well as an evaluable liver biopsy from radiation-exposed liver tissue within 7 days before MRI were retrospectively identified. Planning computed tomography (CT)/dosimetry was merged to the CT-documentation of the liver biopsy and to the MRI. Presence/absence of radiation-induced liver damage (histopathology) and Gd-BOPTA uptake (MRI) as well as the dose applied during brachytherapy at the site of tissue sampling was determined.ResultsFourteen biopsies from eight patients were evaluated. In all cases with histopathological evidence of radiation-induced liver damage (n = 11), no uptake of Gd-BOPTA was seen. In the remaining three, cases no radiation-induced liver damage but Gd-BOPTA uptake was seen. Presence of radiation-induced liver damage and absence of Gd-BOPTA uptake was correlated with a former high-dose exposition.ConclusionsAbsence of hepatobiliary MRI contrast media uptake in radiation-exposed liver parenchyma may indicate radiation-induced liver damage. Confirmatory studies are warranted.

  12. Use of alkaline or enzymatic sample pretreatment prior to characterization of gold nanoparticles in animal tissue by single-particle ICPMS

    DEFF Research Database (Denmark)

    Löschner, Katrin; Brabrand, Myung Suk Jung; Sloth, Jens Jørgen

    2014-01-01

    , not much is known about the applicability of spICPMS for determination of NPs in complex matrices such as biological tissues. In the present study, alkaline and enzymatic treatments were applied to solubilize spleen samples from rats, which had been administered 60-nm gold nanoparticles (Au...... concentration obtained by conventional ICPMS analysis of acid-digested tissue. The recovery of AuNPs from enzymatically digested tissue, however, was approximately four times lower. Spiking experiments of blank spleen samples with AuNPs showed that the lower recovery was caused by an inferior transport...

  13. Basic Histopathological Methods and Breast Lesion Types for Research.

    Science.gov (United States)

    Ouyang, Nengtai; Wang, Lin

    2016-01-01

    The in situ observation on the tissues, such as histopathology, immunohistochemistry (IHC), immunofluorescence (IF), and in situ hybridization (ISH), is one of the most important methods in the biomedical scientific research. In this chapter we introduce the most often used methods-hematoxylin and eosin (H&E) and double IF staining. H&E staining is used for general morphology by which the different pathological types of breast lesions are identified. The double IF staining is often used to study the protein-protein interaction on tissues for signaling mechanisms. This chapter also includes the histopathology of primary or simplified breast lesion types that is essential for applying the above methods and the reclassification of breast cancers by molecular markers.

  14. Activity assays of mammalian thioredoxin and thioredoxin reductase: fluorescent disulfide substrates, mechanisms, and use with tissue samples.

    Science.gov (United States)

    Montano, Sergio J; Lu, Jun; Gustafsson, Tomas N; Holmgren, Arne

    2014-03-15

    Thioredoxin (Trx) is a protein disulfide reductase that, together with nicotinamide adenine dinucleotide phosphate (NADPH) and thioredoxin reductase (TrxR), controls oxidative stress or redox signaling via thiol redox control. Human cytosolic Trx1 has Cys32 and Cys35 as the active site and three additional cysteine residues (Cys62, Cys69, and Cys73), which by oxidation generates inactive Cys62 to Cys69 two-disulfide Trx. This, combined with TrxR with a broad substrate specificity, complicates assays of mammalian Trx and TrxR. We sought to understand the autoregulation of Trx and TrxR and to generate new methods for quantification of Trx and TrxR. We optimized the synthesis of two fluorescent substrates, di-eosin-glutathione disulfide (Di-E-GSSG) and fluorescein isothiocyanate-labeled insulin (FiTC-insulin), which displayed higher fluorescence on disulfide reduction. Di-E-GSSG showed a very large increase in fluorescence quantum yield but had a relatively low affinity for Trx and was also a weak direct substrate for TrxR, in contrast to GSSG. FiTC-insulin was used to develop highly sensitive assays for TrxR and Trx. Reproducible conditions were developed for reactivation of modified Trx, commonly present in frozen or oxidized samples. Trx in cell extracts and tissue samples, including plasma and serum, were subsequently analyzed, showing highly reproducible results and allowing measurement of trace amounts of Trx.

  15. A study of infestation of Alburnoides bipunctatus with Ligula intestinalis in Latian reservoir Dam Lake, Tehran province, Iran: A histopathological stud

    Directory of Open Access Journals (Sweden)

    Iraj Sohrabi Haghdoos

    2011-06-01

    Full Text Available . Objective: The objective of this study was to report infestation of the fish with Ligula intestinalis as a food borne zoonoses and to study its histopathological effects on one Alburnoides bipunctatus in Latian reservoir Dam Lake, Tehran province, Iran. Material and methods: In July 2010, 6 fish (Alburnoides bipunctatus from Latian reservoir Dam Lake, Tehran province, Iran were referred to Laboratory. After autopsy, two of them (Mature and Female were found infested with plerocercoid of Ligula intestinalis based on taxonomical features. For histopathological studies samples were stained with Haematoxylin and Eosin (H&E and observed under light microscopy. Results: Pathological findings showed inflammation of the reservoir adipose tissue (in the loose conjunctive tissue between lipocytes surrounding intestine. Severe atrophy in follicular cells of ovary was evident. In liver cholangiohepastitis with metaplastic hyperplasia was seen. Conclusion: Considering the importance ofLigula intestinalis as a food borne parasite, performance of other comprehensive studies is a necessity.

  16. Biased visualization of hypoperfused tissue by computed tomography due to short imaging duration: improved classification by image down-sampling and vascular models

    Energy Technology Data Exchange (ETDEWEB)

    Mikkelsen, Irene Klaerke; Ribe, Lars Riisgaard; Bekke, Susanne Lise; Tietze, Anna; Oestergaard, Leif; Mouridsen, Kim [Aarhus University Hospital, Center of Functionally Integrative Neuroscience, Aarhus C (Denmark); Jones, P.S.; Alawneh, Josef [University of Cambridge, Department of Clinical Neurosciences, Cambridge (United Kingdom); Puig, Josep; Pedraza, Salva [Dr. Josep Trueta Girona University Hospitals, Department of Radiology, Girona Biomedical Research Institute, Girona (Spain); Gillard, Jonathan H. [University of Cambridge, Department of Radiology, Cambridge (United Kingdom); Warburton, Elisabeth A. [Cambrigde University Hospitals, Addenbrooke, Stroke Unit, Cambridge (United Kingdom); Baron, Jean-Claude [University of Cambridge, Department of Clinical Neurosciences, Cambridge (United Kingdom); Centre Hospitalier Sainte Anne, INSERM U894, Paris (France)

    2015-07-15

    Lesion detection in acute stroke by computed-tomography perfusion (CTP) can be affected by incomplete bolus coverage in veins and hypoperfused tissue, so-called bolus truncation (BT), and low contrast-to-noise ratio (CNR). We examined the BT-frequency and hypothesized that image down-sampling and a vascular model (VM) for perfusion calculation would improve normo- and hypoperfused tissue classification. CTP datasets from 40 acute stroke patients were retrospectively analysed for BT. In 16 patients with hypoperfused tissue but no BT, repeated 2-by-2 image down-sampling and uniform filtering was performed, comparing CNR to perfusion-MRI levels and tissue classification to that of unprocessed data. By simulating reduced scan duration, the minimum scan-duration at which estimated lesion volumes came within 10 % of their true volume was compared for VM and state-of-the-art algorithms. BT in veins and hypoperfused tissue was observed in 9/40 (22.5 %) and 17/40 patients (42.5 %), respectively. Down-sampling to 128 x 128 resolution yielded CNR comparable to MR data and improved tissue classification (p = 0.0069). VM reduced minimum scan duration, providing reliable maps of cerebral blood flow and mean transit time: 5 s (p = 0.03) and 7 s (p < 0.0001), respectively. BT is not uncommon in stroke CTP with 40-s scan duration. Applying image down-sampling and VM improve tissue classification. (orig.)

  17. Digital staining for histopathology multispectral images by the combined application of spectral enhancement and spectral transformation.

    Science.gov (United States)

    Bautista, Pinky A; Yagi, Yukako

    2011-01-01

    In this paper we introduced a digital staining method for histopathology images captured with an n-band multispectral camera. The method consisted of two major processes: enhancement of the original spectral transmittance and the transformation of the enhanced transmittance to its target spectral configuration. Enhancement is accomplished by shifting the original transmittance with the scaled difference between the original transmittance and the transmittance estimated with m dominant principal component (PC) vectors;the m-PC vectors were determined from the transmittance samples of the background image. Transformation of the enhanced transmittance to the target spectral configuration was done using an nxn transformation matrix, which was derived by applying a least square method to the enhanced and target spectral training data samples of the different tissue components. Experimental results on the digital conversion of a hematoxylin and eosin (H&E) stained multispectral image to its Masson's trichrome stained (MT) equivalent shows the viability of the method.

  18. Histopathologic study of eosinophilic bronchointerstitial pneumonia caused by Crenosoma striatum in the hedgehog.

    Science.gov (United States)

    Hoseini, Seyed Mohammad; Youssefi, Mohammad Reza; Mousapour, Aliasghar; Dozouri, Rohollah; Eshkevari, Shahab Ramezanpour; Nikzad, Mohammad; Nikzad, Reza; Omidzahir, Shila

    2014-06-01

    Crenosoma striatum is a species of parasitic nematodes from the family Crenosomatidae responsible for pathologic lung lesions in the hedgehog (Erinaceus europaeus). Infection with C. striatum can cause weight loss, dry cough, and bronchitis. In the present study, hedgehogs killed by road accidents, or trapped and found dead on farms in different parts of Mazandaran province (Iran), were transferred to the laboratory. After dissection, parasite samples collected from the lung were placed into 70% alcohol. After clarification with lactophenol and subsequent staining, the nematodes were identified as C. striatum according to previously published guidelines. For histopathologic examination, lung samples were collected. The tissues were fixed and following routine processing, sections were stained with hematoxylin and eosin. Microscopic diagnoses included hyperemia, eosinophilic bronchointerstitial pneumonia, thickening of the interstitium, and eosinophilic microabscesses in bronchial airways. Eosinophilic pneumonia was characterized by eosinophil and other mononuclear leukocyte infiltration within the lung interstitium. Crenosoma striatum can lead to mortality in hedgehogs.

  19. A Histopathological Study of the so-called Buruli ulcer

    OpenAIRE

    山下, 裕人; 千馬, 正敬; 板倉, 英世

    1980-01-01

    A histopathological study of the so-called "Buruli ulcer" was reported. The patient was a 10-year-old African boy in Kenya suffering from ulcer formation. The ulcer was 10cm in diameter and located over the left hip. It was undermined and covered by overhanged epithelia at the ulcer edge. There was edema or lytic change in the dermis which formed gaps between the epidermis and dermis in adjacent regions. Microscopically, inflammatory cells infiltration, granulation tissue, fibrosis, and sever...

  20. Neuroradiology and histopathology in two cases of adult medulloblastoma.

    Science.gov (United States)

    Romero-Rojas, Alfredo E; Diaz-Perez, Julio A; Raju, Sharat; Lozano-Castillo, Alfonso

    2014-04-01

    Medulloblastoma (MB) is the most common central nervous system neoplasm in children and only rarely presents in the adult population. Recent molecular biology findings have characterized MB as a heterogeneous neoplasm distinguished by well-defined tumour subsets each with specific histologic and molecular features. Available immunohistochemical stains can now be used to differentiate the distinct molecular types of MB. This report analyzed the histopathologic and neuroradiologic features of two new cases of adult MB. Imaging studies in these patients revealed the morphological appearance of high-grade, well-circumscribed heterogeneous tumours with necrosis, located laterally within the posterior cranial fossa. Histopathology of resected samples demonstrated high-grade tumours (WHO grade IV) containing sheets of undifferentiated neural cells with high mitotic activity and evidence of necrosis. The histopathologic and molecular characteristics of these cases of MB are reviewed for potential applications in new molecular methods of imaging.

  1. Histopathology of human laser thermal angioplasty recanalization.

    Science.gov (United States)

    White, R A; White, G H; Vlasak, J; Fujitani, R; Kopchok, G E

    1988-01-01

    Laserprobe thermal-assisted balloon, angioplasty (LTBA) has demonstrated promising initial clinical results in recanalizing stenotic or occluded superficial femoral and popliteal arteries. Over the past year we have obtained six specimens of laserprobe thermal (LT) and LTBA treated total occlusions (avg. length 12 cm) for histopathologic examination from patients who were treated for limb salvage. Three tissue specimens were obtained acutely, and one was obtained at 6, 8, and 13 days, respectively, after laser angioplasty at the time of revision for complications or failed procedures. Serial histologic sections of the treated LT segments demonstrated recanalization of atherosclerotic lesions to approximately 60-70% of the probe diameter. The LT channels were lined by a thin layer of carbonized or coagulated tissue and several layers of cell necrosis. The histology of the thermal injury was similar regardless of whether it was produced by the heated metal cap or by free argon laser energy. Stellate balloon angioplasty fractures were frequently filled with thrombus. Analysis of these human LT and LTBA specimens revealed that the thermal device produces a confined injury through the path of least resistance. Balloon dilatation produces fragmented cracks in the vessel wall, which appear to be more thrombogenic than the carbonized LT surface. With improved guidance methods, LTBA shows potential for continuing development.

  2. Postmenopausal osteoporosis - clinical, biological and histopathological aspects.

    Science.gov (United States)

    Pavel, Oana Roxana; Popescu, Mihaela; Novac, Liliana; Mogoantă, LaurenŢiu; Pavel, LaurenŢiu Petrişor; Vicaş, Răzvan Marius; Trăistaru, Magdalena Rodica

    2016-01-01

    Osteoporosis is one of the most common disorders in postmenopausal women, affecting the quality of life and increasing the risk for fractures in minor traumas. Changes in the bone microarchitecture causes static changes in the body and affects motility. In this study, we analyzed two groups of women, one with physiological menopause and one with surgically induced menopause. The diagnosis of osteoporosis was suspected based on the clinical symptoms and confirmed by assessing bone mineral density by the dual-energy X-ray absorptiometry (DEXA). Comparing some clinical and biological aspects there was noted that a much higher percentage of women with surgically induced menopause exhibited increases in body mass index, changes in serum lipids, cholesterol, triglycerides, blood glucose, serum calcium, magnesemia and osteocalcin. In contrast, no significant differences were observed in the histopathological aspects of bone tissue examined from these two groups. In all patients, there was identified a significant reduction in the number of osteocytes and osteoblasts, the expansion of haversian channels, reducing the number of trabecular bone in the cancellous bone with wide areola cavities often full of adipose tissue, non-homogenous demineralization of both the compact bone and the cancellous bone, atrophy and even absence of the endosteal, and the presence of multiple microfractures. Our study showed that early surgically induced menopause more intensely alters the lipid, carbohydrate and mineral metabolism, thus favoring the onset of osteoporosis.

  3. Phase II Trial of Cetuximab and Conformal Radiotherapy Only in Locally Advanced Pancreatic Cancer with Concurrent Tissue Sampling Feasibility Study.

    Science.gov (United States)

    Rembielak, Agata I; Jain, Pooja; Jackson, Andrew S; Green, Melanie M; Santorelli, Gillian R; Whitfield, Gillian A; Crellin, Adrian; Garcia-Alonso, Angel; Radhakrishna, Ganesh; Cullen, James; Taylor, M Ben; Swindell, Ric; West, Catharine M; Valle, Juan; Saleem, Azeem; Price, Patricia M

    2014-02-01

    Preclinical data have indicated the anti-epidermal growth factor receptor (EGFR) agent cetuximab (Erbitux) as a radiosensitizer in pancreatic cancer, but this has not been specifically addressed in a clinical study. We report the results of an original study initiated in 2007, where cetuximab was tested with radiotherapy (RT) alone in locally advanced pancreatic cancer in a phase II trial (PACER). Patients (n = 21) received cetuximab loading dose (400 mg/m(2)) and weekly dose (250 mg/m(2)) during RT (50.4 Gy in 28 fractions). Toxicity and disease response end point data were prospectively assessed. A feasibility study of on-trial patient blood and skin sampling was incorporated. Treatment was well tolerated, and toxicity was low; most patients (71%) experienced acute toxicities of grade 2 or less. Six months posttreatment, stable local disease was achieved in 90% of evaluable patients, but only 33% were free from metastatic progression. Median overall survival was 7.5 months, and actuarial survival was 33% at 1 year and 11% at 3 years, reflecting swift metastatic progression in some patients but good long-term control of localized disease in others. High-grade acneiform rash (P = .0027), posttreatment stable disease (P = .0059), and pretreatment cancer antigen 19.9 (CA19.9) level (P = .0042) associated with extended survival. Patient skin and blood samples yielded sufficient RNA and good quality protein, respectively. The results indicate that cetuximab inhibits EGFR-mediated radioresistance to achieve excellent local control with minimal toxicity but does not sufficiently control metastatic progression in all patients. Translational studies of patient tissue samples may yield molecular information that may enable individual treatment response prediction.

  4. Exploring the cellular and tissue uptake of nanomaterials in a range of biological samples using multimodal nonlinear optical microscopy

    Science.gov (United States)

    Johnston, Helinor J.; Mouras, Rabah; Brown, David M.; Elfick, Alistair; Stone, Vicki

    2015-12-01

    The uptake of nanomaterials (NMs) by cells is critical in determining their potential biological impact, whether beneficial or detrimental. Thus, investigation of NM internalization by cells is a common consideration in hazard and efficacy studies. There are currently a number of approaches that are routinely used to investigate NM-cell interactions, each of which have their own advantages and limitations. Ideally, imaging modalities used to investigate NM uptake by cells should not require the NM to be labelled (e.g. with fluorophores) to facilitate its detection. We present a multimodal imaging approach employing a combination of label-free microscopies that can be used to investigate NM-cell interactions. Coherent anti-Stokes Raman scattering microscopy was used in combination with either two-photon photoluminescence or four-wave mixing (FWM) to visualize the uptake of gold or titanium dioxide NMs respectively. Live and fixed cell imaging revealed that NMs were internalized by J774 macrophage and C3A hepatocyte cell lines (15-31 μg ml-1). Sprague Dawley rats were exposed to NMs (intratracheal instillation, 62 μg) and NMs were detected in blood and lung leucocytes, lung and liver tissue, demonstrating that NMs could translocate from the exposure site. Obtained data illustrate that multimodal nonlinear optical microscopy may help overcome current challenges in the assessment of NM cellular uptake and biodistribution. It is therefore a powerful tool that can be used to investigate unlabelled NM cellular and tissue uptake in three dimensions, requires minimal sample preparation, and is applicable to live and fixed cells.

  5. A novel method for single sample multi-axial nanoindentation of hydrated heterogeneous tissues based on testing great white shark jaws.

    Science.gov (United States)

    Ferrara, Toni L; Boughton, Philip; Slavich, Eve; Wroe, Stephen

    2013-01-01

    Nanomechanical testing methods that are suitable for a range of hydrated tissues are crucial for understanding biological systems. Nanoindentation of tissues can provide valuable insights into biology, tissue engineering and biomimetic design. However, testing hydrated biological samples still remains a significant challenge. Shark jaw cartilage is an ideal substrate for developing a method to test hydrated tissues because it is a unique heterogeneous composite of both mineralized (hard) and non-mineralized (soft) layers and possesses a jaw geometry that is challenging to test mechanically. The aim of this study is to develop a novel method for obtaining multidirectional nanomechanical properties for both layers of jaw cartilage from a single sample, taken from the great white shark (Carcharodon carcharias). A method for obtaining multidirectional data from a single sample is necessary for examining tissue mechanics in this shark because it is a protected species and hence samples may be difficult to obtain. Results show that this method maintains hydration of samples that would otherwise rapidly dehydrate. Our study is the first analysis of nanomechanical properties of great white shark jaw cartilage. Variation in nanomechanical properties were detected in different orthogonal directions for both layers of jaw cartilage in this species. The data further suggest that the mineralized layer of shark jaw cartilage is less stiff than previously posited. Our method allows multidirectional nanomechanical properties to be obtained from a single, small, hydrated heterogeneous sample. Our technique is therefore suitable for use when specimens are rare, valuable or limited in quantity, such as samples obtained from endangered species or pathological tissues. We also outline a method for tip-to-optic calibration that facilitates nanoindentation of soft biological tissues. Our technique may help address the critical need for a nanomechanical testing method that is applicable

  6. A novel method for single sample multi-axial nanoindentation of hydrated heterogeneous tissues based on testing great white shark jaws.

    Directory of Open Access Journals (Sweden)

    Toni L Ferrara

    Full Text Available Nanomechanical testing methods that are suitable for a range of hydrated tissues are crucial for understanding biological systems. Nanoindentation of tissues can provide valuable insights into biology, tissue engineering and biomimetic design. However, testing hydrated biological samples still remains a significant challenge. Shark jaw cartilage is an ideal substrate for developing a method to test hydrated tissues because it is a unique heterogeneous composite of both mineralized (hard and non-mineralized (soft layers and possesses a jaw geometry that is challenging to test mechanically. The aim of this study is to develop a novel method for obtaining multidirectional nanomechanical properties for both layers of jaw cartilage from a single sample, taken from the great white shark (Carcharodon carcharias. A method for obtaining multidirectional data from a single sample is necessary for examining tissue mechanics in this shark because it is a protected species and hence samples may be difficult to obtain. Results show that this method maintains hydration of samples that would otherwise rapidly dehydrate. Our study is the first analysis of nanomechanical properties of great white shark jaw cartilage. Variation in nanomechanical properties were detected in different orthogonal directions for both layers of jaw cartilage in this species. The data further suggest that the mineralized layer of shark jaw cartilage is less stiff than previously posited. Our method allows multidirectional nanomechanical properties to be obtained from a single, small, hydrated heterogeneous sample. Our technique is therefore suitable for use when specimens are rare, valuable or limited in quantity, such as samples obtained from endangered species or pathological tissues. We also outline a method for tip-to-optic calibration that facilitates nanoindentation of soft biological tissues. Our technique may help address the critical need for a nanomechanical testing method

  7. Molecular Histopathology by Spectrally Reconstructed Nonlinear Interferometric Vibrational Imaging

    Science.gov (United States)

    Chowdary, Praveen D.; Jiang, Zhi; Chaney, Eric J.; Benalcazar, Wladimir A.; Marks, Daniel L.; Gruebele, Martin; Boppart, Stephen A.

    2011-01-01

    Sensitive assays for rapid quantitative analysis of histologic sections, resected tissue specimens, or in situ tissue are highly desired for early disease diagnosis. Stained histopathology is the gold standard but remains a subjective practice on processed tissue taking from hours to days. We describe a microscopy technique that obtains a sensitive and accurate color-coded image from intrinsic molecular markers. Spectrally reconstructed nonlinear interferometric vibrational imaging can differentiate cancer versus normal tissue sections with greater than 99% confidence interval in a preclinical rat breast cancer model and define cancer boundaries to ±100 μm with greater than 99% confidence interval, using fresh unstained tissue sections imaged in less than 5 minutes. By optimizing optical sources and beam delivery, this technique can potentially enable real-time point-of-care optical molecular imaging and diagnosis. PMID:21098699

  8. American Badger Study : Badger Tissue Collection Check Sheets, Batched Samples Tracking Logs & Wildlife Sample Collection Forms : Rocky Mountain Arsenal NWR : 1994

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This record contains various data sheets related to the American Badger Study conducted at the Rocky Mountain Arsenal National Wildlife Refuge; badger tissue...

  9. Next-generation sequencing of RNA and DNA isolated from paired fresh-frozen and formalin-fixed paraffin-embedded samples of human cancer and normal tissue.

    Directory of Open Access Journals (Sweden)

    Jakob Hedegaard

    Full Text Available Formalin-fixed, paraffin-embedded (FFPE tissues are an invaluable resource for clinical research. However, nucleic acids extracted from FFPE tissues are fragmented and chemically modified making them challenging to use in molecular studies. We analysed 23 fresh-frozen (FF, 35 FFPE and 38 paired FF/FFPE specimens, representing six different human tissue types (bladder, prostate and colon carcinoma; liver and colon normal tissue; reactive tonsil in order to examine the potential use of FFPE samples in next-generation sequencing (NGS based retrospective and prospective clinical studies. Two methods for DNA and three methods for RNA extraction from FFPE tissues were compared and were found to affect nucleic acid quantity and quality. DNA and RNA from selected FFPE and paired FF/FFPE specimens were used for exome and transcriptome analysis. Preparations of DNA Exome-Seq libraries was more challenging (29.5% success than that of RNA-Seq libraries, presumably because of modifications to FFPE tissue-derived DNA. Libraries could still be prepared from RNA isolated from two-decade old FFPE tissues. Data were analysed using the CLC Bio Genomics Workbench and revealed systematic differences between FF and FFPE tissue-derived nucleic acid libraries. In spite of this, pairwise analysis of DNA Exome-Seq data showed concordance for 70-80% of variants in FF and FFPE samples stored for fewer than three years. RNA-Seq data showed high correlation of expression profiles in FF/FFPE pairs (Pearson Correlations of 0.90 +/- 0.05, irrespective of storage time (up to 244 months and tissue type. A common set of 1,494 genes was identified with expression profiles that were significantly different between paired FF and FFPE samples irrespective of tissue type. Our results are promising and suggest that NGS can be used to study FFPE specimens in both prospective and retrospective archive-based studies in which FF specimens are not available.

  10. Comparison between cytology and histopathology to evaluate subclinical endometritis in dairy cows.

    Science.gov (United States)

    Bogado Pascottini, Osvaldo; Hostens, Miel; Dini, Pouya; Vandepitte, Jan; Ducatelle, Richard; Opsomer, Geert

    2016-10-01

    The aim of the present study was to compare endometrial cytology with histopathology to diagnose subclinical endometritis (SCE) in dairy cows. Endometrial cytology samples were collected from Holstein-Friesian cows (n = 32) just before slaughtering. Half of them were obtained by in vivo cytobrush (IV-CB), whereas the other half by in vivo low-volume lavage (IV-LVL). After slaughtering, reproductive tracts were collected, and the endometrium was sampled at eight locations. At each location, both a ex vivo cytobrush sample (EV-CB) and a tissue sample for histopathologic examination were taken. In the histopathology slides, polymorphonuclear (PMN) cell counts were differentiated as PMN cells in direct contact with the epithelial cells of the endometrium (PMN-EP), and PMN cells present in the deeper stratum compactum (PMN-SC). Summation of both countings was referred to as PMN-total. Pearson's correlation and Cohen's kappa coefficient were used to assess the correlation and agreement between both sampling methods (in vivo cytology [IV-CB and IV-LVL] with EV-CB and PMN-total). A Poisson mixed effect model was used to analyze the PMN cells' distribution. The prevalence of SCE was 18.75% (n = 6/32) for in vivo cytology. The SCE prevalence based on EV-CB analyses and on the assessment of PMN-total was determined both at the sample (n = 256) as well as at the cow level (n = 32): EV-CB 25% (n = 64/256) and 35.5% (n = 12/32), and PMN-total 37.11% (n = 95/256) and 59.38% (n = 19/32). Correlation and agreement between IV-CB and EV-CB were r = 0.81 and k = 0.97, whereas between IV-CB and PMN-total r = 0.15 and k = 0.23, respectively. In vivo low-volume lavage correlation and agreement were r = 0.52 and k = 0.66 with EV-CB, and r = 0.45 and k = 0.44 with PMN-total. Moreover, correlation and agreement between EV-CB and PMN-total were r = 0.60 and k = 0.50, respectively. More PMN cells (P histopathology, rendering the latter as a more

  11. Molecular identification and histopathological study of natural Streptococcus agalactiae infection in hybrid tilapia (Oreochromis niloticus)

    Science.gov (United States)

    Laith, AA; Ambak, Mohd Azmi; Hassan, Marina; Sheriff, Shahreza Md.; Nadirah, Musa; Draman, Ahmad Shuhaimi; Wahab, Wahidah; Ibrahim, Wan Nurhafizah Wan; Aznan, Alia Syafiqah; Jabar, Amina; Najiah, Musa

    2017-01-01

    Aim: The main objective of this study was to emphasize on histopathological examinations and molecular identification of Streptococcus agalactiae isolated from natural infections in hybrid tilapia (Oreochromis niloticus) in Temerloh Pahang, Malaysia, as well as to determine the susceptibility of the pathogen strains to various currently available antimicrobial agents. Materials and Methods: The diseased fishes were observed for variable clinical signs including fin hemorrhages, alterations in behavior associated with erratic swimming, exophthalmia, and mortality. Tissue samples from the eyes, brain, kidney, liver, and spleen were taken for bacterial isolation. Identification of S. agalactiae was screened by biochemical methods and confirmed by VITEK 2 and 16S rRNA gene sequencing. The antibiogram profiling of the isolate was tested against 18 standard antibiotics included nitrofurantoin, flumequine, florfenicol, amoxylin, doxycycline, oleandomycin, tetracycline, ampicillin, lincomycin, colistin sulfate, oxolinic acid, novobiocin, spiramycin, erythromycin, fosfomycin, neomycin, gentamycin, and polymyxin B. The histopathological analysis of eyes, brain, liver, kidney, and spleen was observed for abnormalities related to S. agalactiae infection. Results: The suspected colonies of S. agalactiae identified by biochemical methods was observed as Gram-positive chained cocci, β-hemolytic, and non-motile. The isolate was confirmed as S. agalactiae by VITEK 2 (99% similarity), reconfirmed by 16S rRNA gene sequencing (99% similarity) and deposited in GenBank with accession no. KT869025. The isolate was observed to be resistance to neomycin and gentamicin. The most consistent gross findings were marked hemorrhages, erosions of caudal fin, and exophthalmos. Microscopic examination confirmed the presence of marked congestion and infiltration of inflammatory cell in the eye, brain, kidney, liver, and spleen. Eye samples showed damage of the lens capsule, hyperemic and

  12. [Update on protein analysis of fixed tissues].

    Science.gov (United States)

    Becker, K-F; Berg, D; Malinowsky, K; Wolff, C; Ergin, B; Meding, S; Walch, A; Höfler, H

    2010-10-01

    Tissue samples have been routinely used for decades to distinguish healthy from diseased tissue in histopathological characterization. While nucleic acid-based methodologies have been successfully in use for many years, protein-based techniques, in contrast, are at a very early stage (with the exception of immunohistochemistry). One reason for this delay may be that the scientific community has long thought that formalin-fixed and paraffin embedded (FFPE) tissues are unfit for protein analysis. However, recent reports demonstrate that many protein methods that are routinely used for frozen tissues can also be applied for FFPE tissues, including Western blot, protein microarray, matrix-assisted laser desorption/ionization (MALDI) imaging and 2D gel electrophoresis. The present article provides an overview of recent developments in this field, focussing particular attention on quantitative analysis and high throughput technologies that have the potential to be integrated into the routine workflow of clinical pathology laboratories.

  13. Confidentiality, informed consent, and children's participation in research involving stored tissue samples: interviews with medical professionals from the Middle East.

    Science.gov (United States)

    Alahmad, Ghiath; Al Jumah, Mohammed; Dierickx, Kris

    2015-01-01

    Ethical issues regarding research biobanks continue to be a topic of intense debate, especially issues of confidentiality, informed consent, and child participation. Although considerable empirical literature concerning research biobank ethics exists, very little information is available regarding the opinions of medical professionals doing genetics research from the Middle East, especially Arabic speaking countries. Ethical guidelines for research biobanks are critically needed as some countries in the Middle East are starting to establish national research biobanks. Islam is the dominant religion in these countries, and it affects people's behavior and influences their positions. Moreover, communities in these countries enjoy a set of customs, traditions and social norms, and have social and familial structures that must be taken into account when developing research policies. We interviewed 12 medical professionals from the Middle East currently working with stored tissue samples to document their opinions. We found general agreement. Participants' primary concerns were similar to the views of researchers internationally. Since children tend to represent a high percentage of Middle Eastern populations, and because children's bodies are not just small adult bodies, the interviewed professionals strongly believed that it is imperative to include children in biobank research. Participants generally believed that protecting confidentiality is socially very important and that informed consent/assent must be obtained from both adult and child participants. This study provides a starting point for additional studies.

  14. 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.

  15. [Histopathological evaluation of the meniscus].

    Science.gov (United States)

    Fisseler-Eckhoff, A; Müller, K-M

    2011-05-01

    Menisci fulfill many functions within the complex biomechanics of the knee joint. In the case of meniscus lesions, sparing arthroscopic resection and surgical refixation are the treatments of choice. In terms of diagnosis, this means in general that histopathologic diagnostics are carried out on detached meniscus fragments of between 5 mm and 2 cm in size. A good knowledge of physiologically possible cellular and fibrous histological meniscus damage, as opposed to nonphysiological change regarded as normal with respect to age, is essential for diagnostic meniscus evaluation. The clinician expects clear statements from the pathologist regarding the severity of previous or secondary degenerative meniscus damage, the age and type of traumatic tears, and an appraisal of the relationship between trauma and meniscus damage from an insurance point of view. Close cooperation between the clinician and the pathologist allows for a fast and unambiguous correlation of anamnesis, the clinical picture, and morphological reporting such that problematic insurance cases can be clarified quickly.

  16. Application of a Liquid Extraction Based Sealing Surface Sampling Probe for Mass Spectrometric Analysis of Dried Blood Spots and Mouse Whole-Body Thin Tissue Sections

    Energy Technology Data Exchange (ETDEWEB)

    Van Berkel, Gary J [ORNL; Kertesz, Vilmos [ORNL

    2009-01-01

    The utility of a liquid extraction based sealing surface sampling probe (SSSP) for the direct mass spectrometric analysis of targeted drugs and metabolites in dried blood spots (DBSs) and whole mouse thin tissue sections was demonstrated. The accuracy and precision for the quantitative analysis of a minimum of 50 ng/mL sitamaquine or acetaminophen in DBSs on paper were well within the required 15% dictated by internationally recognized acceptance criteria for assay validations. Analysis of whole-body mouse thin tissue sections from animals dosed with propranolol, adhered to an adhesive tape substrate, provided semi-quantitative information for propranolol and its hydroxyproranolol glucuronide metabolite within specific organs of the tissue. The relative abundances recorded for the two compounds in the brain, lung, kidney and liver were in nominal agreement with previously reported amounts based on analysis using a liquid microjunction surface sampling probe (LMJ-SSP), and whole-body autoradiography (WBA) and HPLC-MS analysis. The ability to sample and analyze from tape-adhered tissue samples, which are generally employed in WBA analysis, presents the possibility of consecutive WBA and SSSP-MS analysis of the same tissue section. This would facilitate assignment, and possibly quantitation, of the different molecular forms of total drug related material detected in the WBA analysis. The flexibility to sample larger or smaller spot sizes, alternative probe sealing mechanisms, and a reduction in internal volumes and associated sample carryover issues will be among the first simple improvements necessary to make the SSSP-MS method a practical DBS and/or thin tissue section analysis tool or to expand its use to other surface sampling applications.

  17. Effect of topical anaesthetics on interstitial colloid osmotic pressure in human subcutaneous tissue sampled by wick technique.

    Directory of Open Access Journals (Sweden)

    Hans Jørgen Timm Guthe

    Full Text Available OBJECTIVES: To measure colloid osmotic pressure in interstitial fluid (COP(i from human subcutaneous tissue with the modified wick technique in order to determine influence of topical application of anaesthetics, dry vs. wet wick and implantation time on COP(i. MATERIAL AND METHODS: In 50 healthy volunteers interstitial fluid (IF was collected by subcutaneous implantation of multi-filamentous nylon wicks. Study subjects were allocated to two groups; one for comparing COP(i obtained from dry and saline soaked wicks, and one for comparing COP(i from unanaesthetized skin, and skin after application of a eutectic mixture of local anaesthetic (EMLA®, Astra Zeneca cream. IF was sampled from the skin of the shoulders, and implantation time was 30, 60, 75, 90 and 120 min. Colloid osmotic pressure was measured with a colloid osmometer. Pain assessment during the procedure was compared for EMLA cream and no topical anaesthesia using a visual analogue scale (VAS in a subgroup of 10 subjects. RESULTS: There were no significant differences between COP(i obtained from dry compared to wet wicks, except that the values after 75 and 90 min. were somewhat higher for the dry wicks. Topical anaesthesia with EMLA cream did not affect COP(i values. COP(i decreased from 30 to 75 min. of implantation (23.2 ± 4.4 mmHg to 19.6 ± 2.9 mmHg, p = 0.008 and subsequently tended to increase until 120 min. EMLA cream resulted in significant lower VAS score for the procedure. CONCLUSION: COP(i from subcutaneous tissue was easily obtained and fluid harvesting was well tolerated when topical anaesthetic was used. The difference in COP(i assessed by dry and wet wicks between 75 min. and 90 min. of implantation was in accordance with previous reports. The use of topical analgesia did not influence COP(i and topical analgesia may make the wick technique more acceptable for subjects who dislike technical procedures, including children. TRIAL REGISTRATION: ClinicalTrials.gov NCT

  18. Histological observations in the Hawaiian reef coral, Porites compressa, affected by Porites bleaching with tissue loss

    Science.gov (United States)

    Sudek, M.; Work, T.M.; Aeby, G.S.; Davy, S.K.

    2012-01-01

    The scleractinian finger coral Porites compressa is affected by the coral disease Porites bleaching with tissue loss (PBTL). This disease initially manifests as bleaching of the coenenchyme (tissue between polyps) while the polyps remain brown with eventual tissue loss and subsequent algal overgrowth of the bare skeleton. Histopathological investigation showed a loss of symbiont and melanin-containing granular cells which was more pronounced in the coenenchyme than the polyps. Cell counts confirmed a 65% reduction in symbiont density. Tissue loss was due to tissue fragmentation and necrosis in affected areas. In addition, a reduction in putative bacterial aggregate densities was found in diseased samples but no potential pathogens were observed.

  19. Evaluation of endogenous control genes for gene expression studies across multiple tissues and in the specific sets of fat- and muscle-type samples of the pig.

    Science.gov (United States)

    Gu, Y R; Li, M Z; Zhang, K; Chen, L; Jiang, A A; Wang, J Y; Li, X W

    2011-08-01

    To normalize a set of quantitative real-time PCR (q-PCR) data, it is essential to determine an optimal number/set of housekeeping genes, as the abundance of housekeeping genes can vary across tissues or cells during different developmental stages, or even under certain environmental conditions. In this study, of the 20 commonly used endogenous control genes, 13, 18 and 17 genes exhibited credible stability in 56 different tissues, 10 types of adipose tissue and five types of muscle tissue, respectively. Our analysis clearly showed that three optimal housekeeping genes are adequate for an accurate normalization, which correlated well with the theoretical optimal number (r ≥ 0.94). In terms of economical and experimental feasibility, we recommend the use of the three most stable housekeeping genes for calculating the normalization factor. Based on our results, the three most stable housekeeping genes in all analysed samples (TOP2B, HSPCB and YWHAZ) are recommended for accurate normalization of q-PCR data. We also suggest that two different sets of housekeeping genes are appropriate for 10 types of adipose tissue (the HSPCB, ALDOA and GAPDH genes) and five types of muscle tissue (the TOP2B, HSPCB and YWHAZ genes), respectively. Our report will serve as a valuable reference for other studies aimed at measuring tissue-specific mRNA abundance in porcine samples.

  20. Weakly supervised histopathology cancer image segmentation and classification.

    Science.gov (United States)

    Xu, Yan; Zhu, Jun-Yan; Chang, Eric I-Chao; Lai, Maode; Tu, Zhuowen

    2014-04-01

    Labeling a histopathology image as having cancerous regions or not is a critical task in cancer diagnosis; it is also clinically important to segment the cancer tissues and cluster them into various classes. Existing supervised approaches for image classification and segmentation require detailed manual annotations for the cancer pixels, which are time-consuming to obtain. In this paper, we propose a new learning method, multiple clustered instance learning (MCIL) (along the line of weakly supervised learning) for histopathology image segmentation. The proposed MCIL method simultaneously performs image-level classification (cancer vs. non-cancer image), medical image segmentation (cancer vs. non-cancer tissue), and patch-level clustering (different classes). We embed the clustering concept into the multiple instance learning (MIL) setting and derive a principled solution to performing the above three tasks in an integrated framework. In addition, we introduce contextual constraints as a prior for MCIL, which further reduces the ambiguity in MIL. Experimental results on histopathology colon cancer images and cytology images demonstrate the great advantage of MCIL over the competing methods.

  1. The Histopathological Spectrum of Pyogenic Granuloma: A Case Series

    Directory of Open Access Journals (Sweden)

    Vinay Marla

    2016-01-01

    Full Text Available Background. Pyogenic granuloma is a reactive tumor-like lesion commonly affecting the oral cavity. These lesions usually appear as localized solitary nodule with a sessile or pedunculated base and colour varying from red, purplish, or pink, depending on the vascularity of the lesion. Pyogenic granuloma shows predilection for gingiva and is usually slow growing, but at times it shows rapid growth. The natural course of this lesion can be categorized into three distinct phases, namely, (i cellular phase, (ii capillary phase/vascular phase, and (iii involutionary phase. Histopathologically, pyogenic granuloma is classified into lobular capillary hemangioma (LCH and non-lobular capillary hemangioma (non-LCH. Case Presentation. In this series, four cases (varied age groups and both genders of pyogenic granuloma showing varying histopathological presentation in relation to its clinical course have been described. The lesion in its early phase reveals diffuse endothelial cells, with few budding into capillaries. Among the capillary phase, the LCH type shows numerous blood vessels organized into lobular aggregates whereas the non-LCH type does not show any such organization and resembles granulation tissue. The involutionary phase shows healing of the lesion and is characterized by extensive fibrosis in the connective tissue. Conclusion. In conclusion, knowledge of the various histopathological presentation of this lesion is necessary for proper identification.

  2. Pathologic change of adjacent tissue of breast cancer (A histopathological analysis of 95 cases)%乳腺癌癌旁病变与乳腺癌的关系——附95例病理组织学分析

    Institute of Scientific and Technical Information of China (English)

    赵庆丽; 杜延泽

    2001-01-01

    Objective To study the relationship between pathologic change near breast cancer and breast cancer. Methods To observe the morphological manifestation of histopathology concerning pathologic change of adjacent tissue of breast cancer.Results A typical hyperplasia and breast cancer are closely related in cyclomastopathy, and with the increase of heterotype in its degrees the risk factor of pathologic change near breast cancer will accordingly increase.Conclusion Cyctomastopathy atypical hyperplasia was regarded as an important pathologic change was breast cancer, and it can develop carcinoma in situ under given conditions.%目的探讨乳腺癌癌旁病变与乳腺癌的关系。方法观察乳腺癌癌旁病变的病理组织学形态表现。结果乳腺增生症中不典型增生与乳腺癌关系密切,且随异型程度增加,癌变危险性随之增高。结论乳腺增生症中不典型增生为重要癌旁病变,在一定条件下可发展成原位癌。

  3. Selective culturing and genus-specific PCR detection for identification of Aeromonas in tissue samples to assist the medico-legal diagnosis of death by drowning.

    Science.gov (United States)

    Huys, Geert; Coopman, Vera; Van Varenbergh, Dirk; Cordonnier, Jan

    2012-09-10

    The detection of autochthonous aquatic bacteria in tissue samples from drowning cases is increasingly considered as an alternative approach to assist the medico-legal diagnosis of death by drowning. Bacteria belonging to the genus Aeromonas may be suitable candidates for this application as they are ubiquitous in natural aquatic environments but are generally not part of the human microbiota. The research aims of this study were (i) to develop a sensitive, specific and rapid screening and confirmation method for Aeromonas species in tissue samples and (ii) to evaluate aseptic sternal puncture as a post-mortem sample technique and bone marrow as an alternative matrix to provide evidence of death by drowning. The presence of Aeromonas in tissue samples was verified by cultivation using the selective media Ampicillin Dextrin Agar (ADA) and Ryan's Aeromonas Medium. The use of ADA medium was found most optimal for the sensitive, inexpensive and quick detection of aeromonads in human tissue samples. Positive culture plates were confirmed by harvesting all colonies for DNA extraction and subsequent PCR amplification using Aeromonas genus-specific primers. Aeromonads were detected in lung swab, blood and bone marrow of drowned bodies (n=3), but were negative in these three matrices for all negative controls (n=90) tested. Bone marrow proved to be a suitable alternative matrix and can be sampled post-mortem by an aseptic sternal puncture. In conclusion, this study confirms previous indications that aeromonads in cultures from blood of water bodies can be considered a potential marker for drowning. Given the fact that the number of immersed bodies (drowned and non-drowned) included in this study is statistically not significant, however, more tissue samples need to be investigated to confirm the validity of these methods to aid the diagnosis of death by wet drowning.

  4. Histopathological effects of doxorubicin on pancreas in male albino rats

    Directory of Open Access Journals (Sweden)

    I.A. Ali

    2015-06-01

    Full Text Available The aim of this study was to investigate the histopathological side effects of doxorubicin on pancreas tissue in male albino rats Rattus norvegicus. This study were used 55 adult rats (2.5-3.5 month of age. The rats divided into two groups, the first group include (35 rats. The second group were (20 rats. Microscopial examination of pancreas lesion demonstrated oedema around the acini, swelling of the epithelial cells of acini, occurance of cystic fibrosis (mucoviscidosis at the concentration of (4,5 mg/kg of body weight ,occurrence of small islets that form of few cells and exocrine-endocrine transformation. There were thickness in the walls of blood vessels, thrombus, congestion of blood vessels, we conclude, that doxorubicin had histopathological effect on pancreas in sub-acute doses more than chronic doses.

  5. Primary adenocarcinoma of ureter: A rare histopathological variant

    Directory of Open Access Journals (Sweden)

    Prekshi Chaudhary

    2016-01-01

    Full Text Available Primary carcinoma of ureter is an uncommon malignancy. Of which, mostly are transitional cell carcinomas followed by squamous cell carcinomas and adenocarcinomas being the rarest histopathology encountered. We report a case of adenocarcinoma ureter in a middle-aged male along with its clinical scenario. A 62-year-old male, presented with complaints of lower urinary tract symptoms. Computerized tomography urogram showed a soft tissue lesion at the right ureterovesical junction. Cystoscopic biopsy reported villous adenoma. Diethylene triamine pentaacetic acid scan reported nonfunctioning right kidney. He underwent laparoscopic right nephroureterectomy, and histopathology reported adenocarcinoma of the right lower third of ureter, with positive distal and close radial margins. The patient received external beam radiation to the postoperative bed and lymph nodes, and he is disease-free till date.

  6. [Histopathology of strobilocercosis found in the livers of white mouse.].

    Science.gov (United States)

    Aydin, Nasuhi Engin; Miman, Ozlem; Gül, Mehmet; Daldal, Nilgün

    2010-01-01

    The adult form of Taenia taeniaeformis is found in the intestine of the cat and cheetah. The larva form is called Strobilocercus fasciolaris and is found in rodents such as mice and rats. Our objective was to draw attention to that rare zoonosis, since it has already been reported in the literature as strobilocercosis in humans. During an experimental animal study conducted at Inonu University, some unexpected cystic formations were found in the livers of nine 6-8-month-old healthy white mice, which affected the conducted study negatively. These cystic formations were examined histopathologically. Prepared sections were stained with haemotoxylin eosin, periodic acid-Schiff and Masson trichrome stains, and examined by light microscopy. Strobilocercus fasciolaris larvae that curled towards cyst cavity and their hooks were seen. Plasma cells, macrophage, focus of eosinophilic infiltration and fibroblastic connective tissue were simultaneous found. In this paper, histopathological changes in intermediate hosts caused by Strobilocercus fasciolaris and other cestod larvae have been discussed.

  7. Imaging and histopathological aspects in aseptic osteonecrosis of the femoral head.

    Science.gov (United States)

    Trăistaru, Magdalena Rodica; Kamal, Diana; Kamal, Kamal Constantin; Rogoveanu, Otilia Constantina; Popescu, Mihai; Bondari, Simona; Alexandru, Dragoş Ovidiu; Ionovici, Nina; Grecu, Dan Cristian

    2015-01-01

    Aseptic osteonecrosis causes various clinical manifestations, depending on its location, but has in common a histopathological and radiological substrate. Aseptic osteonecrosis of the femoral head is a condition whose pathogenesis remains unclear despite many theories developed so far, and the discovery of numerous risk factors. The objective of this study is to emphasize the role of imaging techniques and correlating histology and immunohistochemistry methods in order to more accurately stage the disease. This retrospective study was performed on a total of 103 patients with clinical and radiological suspicion of unilateral or bilateral osteonecrosis. For the diagnosis criteria, we used clinical information, pelvic X-ray images, magnetic resonance imaging (MRI) or computed tomography (CT). For the inclusion of patients in a disease stage, we used the Association Research Circulation Osseous (ARCO) classification system. For patients diagnosed at an advanced stage, who underwent hip arthroplasty, we harvested biological material necessary for the histopathological study. There were differences in the appearance and extent of the lesion on the histological samples compared to macroscopic examination and even those obtained through imaging means, particularly for patients in evolutionary stage III. Aspects such as the extension of the area of fibrosis, bone tissue remodeling, the density of the newly formed vascular network and degree of impairment of the cartilage, are determined more accurately using histology and immunohistochemistry techniques. Before classifying patients in a certain stage, after correlating clinical and imaging data, histopathological aspects have to be considered, particularly in patients in stages III and IV, in which total hip arthroplasty could be delayed.

  8. Comparison of two histopathologic methods for evaluating subcutaneous reaction to mineral trioxide aggregate

    Science.gov (United States)

    Lotfi, Mehrdad; Moradzadeh, Monir; Aghbali, Amirala; Rahimi, Saeed; Saghiri, Mohammadali; Zand, Vahid; Mehdipour, Masoumeh; Ranjkesh, Bahram; Doosti, Sirvan

    2012-01-01

    Objectives: One of the most important factors for suitable materials for pulp therapy is biocompatibility. Two histopathologic methods of Cox and Federation Dentaire International (FDI) were used to evaluate inflammation. In Cox method, density of inflammatory cells, tissue reactions like fibrosis, vascular responses like congestion and fibrin extravasation have been used to evaluate inflammatory reactions. The aim of this study was to compare the accuracy of pathologists’ interpretations using two different methods. Study design: Three pathologists observed the degree of inflammation in 225 histopathologic sections. These sections showed inflammation in subcutaneous connective tissue of rats adjacent to polyethylene tubes, filled with white or gray mineral trioxide aggregate. Empty tubes served as controls. Samples were harvested after 7-, 15-, 30-, 60-, and 90-days. All pathologists examined the sections under a light microscope (Carl Zeiss, Oberkochen, Germany) at ×400 magnifications. Chi-Square test was used to evaluate the difference between inflammation grades when one pathologist used two methods. Cohen’s Kappa value was used to measure agreement of three pathologists to recognize the degrees of inflammations when using one of the methods. Results: There were no significant differences between the two methods when one of the pathologist used these methods to report the degree of inflammation (p=0.054). However, two other pathologists reported significant differences between two methods (p=0.005, p=0.001). In the FDI method, there was an acceptable agreement between first and second, and first and third pathologist in terms of the degree of inflammation, and intermediate agreement existed between the second and third pathologist. With the Cox method, no agreement among the pathologists could be found. Conclusion: The results of three pathologists in terms of rating inflammation with the FDI method showed better agreement than with the Cox method

  9. 42 CFR 493.1273 - Standard: Histopathology.

    Science.gov (United States)

    2010-10-01

    ... 42 Public Health 5 2010-10-01 2010-10-01 false Standard: Histopathology. 493.1273 Section 493.1273 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES... Systems § 493.1273 Standard: Histopathology. (a) As specified in § 493.1256(e)(3), fluorescent...

  10. Value of histopathologic examination of uterine products after first-trimester miscarriage.

    Science.gov (United States)

    Alsibiani, Sharifa Ali

    2014-01-01

    The main rationale of routine histopathologic examination of products after first-trimester miscarriages is to detect an ectopic pregnancy or a molar pregnancy, which require further management. An alternative approach is to examine the products only when there is a definite indication. As there is no agreement, we aimed to study whether routine histopathological examination of tissues obtained after first-trimester miscarriage is of any clinical value in our populations. Medical records of all (558) patients with a diagnosis of first-trimester miscarriage over 4 years (2007-2010) at King Abdulaziz University Hospital, Jeddah, Saudi Arabia, were reviewed. Histopathologic examination confirmed products of conception in 537 (96.2%) patients, no products of conception in 17 (3%) patients, molar pregnancy in 2 (0.4%) patients, and decidual tissues without chorionic villi (Arias-Stella reaction) in 2 (0.4%) patients. After clinical correlation, only one unsuspected partial molar pregnancy was diagnosed by histopathology examination. Conclusion is that it does not appear reasonable to perform histopathological examination routinely after all first-trimester miscarriages in our studied population. We recommend that histopathological examination be performed in select instances: when the diagnosis is uncertain, when fewer tissues have been obtained during surgery, when unexpected pathology was seen, when ultrasound suggests a molar pregnancy, or when patients are considered at high risk for trophoblastic disease.

  11. Carotid Catheterization and Automated Blood Sampling Induce Systemic IL-6 Secretion and Local Tissue Damage and Inflammation in the Heart, Kidneys, Liver and Salivary Glands in NMRI Mice

    DEFF Research Database (Denmark)

    Teilmann, Anne Charlotte; Rozell, Björn; Kalliokoski, Otto

    2016-01-01

    Automated blood sampling through a vascular catheter is a frequently utilized technique in laboratory mice. The potential immunological and physiological implications associated with this technique have, however, not been investigated in detail. The present study compared plasma levels of the cyt......Automated blood sampling through a vascular catheter is a frequently utilized technique in laboratory mice. The potential immunological and physiological implications associated with this technique have, however, not been investigated in detail. The present study compared plasma levels...... of the cytokines IL-1β, IL-2, IL-6, IL-10, IL-17A, GM-CSF, IFN-γ and TNF-α in male NMRI mice that had been subjected to carotid artery catheterization and subsequent automated blood sampling with age-matched control mice. Body weight and histopathological changes in the surgical area, including the salivary glands......, the heart, brain, spleen, liver, kidneys and lungs were compared. Catheterized mice had higher levels of IL-6 than did control mice, but other cytokine levels did not differ between the groups. No significant difference in body weight was found. The histology revealed inflammatory and regenerative (healing...

  12. Human breast cancer in vitro: matching histo-pathology with small-angle x-ray scattering and diffraction enhanced x-ray imaging

    Science.gov (United States)

    Fernández, Manuel; Keyriläinen, Jani; Serimaa, Ritva; Torkkeli, Mika; Karjalainen-Lindsberg, Marja-Liisa; Leidenius, Marjut; von Smitten, Karl; Tenhunen, Mikko; Fiedler, Stefan; Bravin, Alberto; Weiss, Thomas M.; Suortti, Pekka

    2005-07-01

    Twenty-eight human breast tumour specimens were studied with small-angle x-ray scattering (SAXS), and 10 of those were imaged by the diffraction enhanced x-ray imaging (DEI) technique. The sample diameter was 20 mm and the thickness 1 mm. Two examples of ductal carcinoma are illustrated by histology images, DEI, and maps of the collagen d-spacing and scattered intensity in the Porod regime, which characterize the SAXS patterns from collagen-rich regions of the samples. Histo-pathology reveals the cancer-invaded regions, and the maps of the SAXS parameters show that in these regions the scattering signal differs significantly from scattering by the surrounding tissue, indicating a degradation of the collagen structure in the invaded regions. The DEI images show the borders between collagen and adipose tissue and provide a co-ordinate system for tissue mapping by SAXS. In addition, degradation of the collagen structure in an invaded region is revealed by fading contrast of the DEI refraction image. The 28 samples include fresh, defrosted tissue and formalin-fixed tissue. The d-values with their standard deviations are given. In the fresh samples there is a systematic 0.76% increase of the d-value in the invaded regions, averaged over 11 samples. Only intra-sample comparisons are made for the formalin-fixed samples, and with a long fixation time, the difference in the d-value stabilizes at about 0.7%. The correspondence between the DEI images, the SAXS maps and the histo-pathology suggests that definitive information on tumour growth and malignancy is obtained by combining these x-ray methods.

  13. Histopathological effects of nanosilver (Ag-NPs in liver after dermal exposure during wound healing

    Directory of Open Access Journals (Sweden)

    Mohammad Saeed Heydarnejad

    2014-04-01

    Full Text Available   Objective(s: With the advent of nanotechnology, significant progress has been made in the area of nanoscale materials such as nanosilver (Ag-Nps. These nanoparticles have a wide range of applications and been used for antimicrobial purposes for more than a century. However, little attention has been paid to the toxicity of nanosilver wound dressing. This study was designed to investigate the possible histopathological toxicity of Ag-NPs in liver of mice during wound healing.     Materials and Methods:   A group of 50 female BALB/c mice of about 8 weeks were randomly divided into two groups: Ag-NPs and control groups (n=25. After creating similar wound on the backs of all animals, the wound bed was treated in Ag-NPs group, with a volume of 50 microliters of the nanosilver solution (10ppm ,and in control group, with the same amount of distilled water. The experiment lasted for 14 days. Histopathaological samplings of liver were conducted on days 2, 7 and 14 of the experiment.   Results: Histopathological studies demonstrated time-dependent changes in mice liver treated with Ag-NPs compared to control group. Some changes include dilation in central venous, hyperemia, cell swelling, increase of Kupffer and inflammatory cells. Conclusion: This study suggests that use of nanosilver for wound healing may cause a mild toxicity, as indicated by time-dependent toxic responses in liver tissue. However, this issue will have to be considered more extensively in further studies.

  14. Fish biomarkers for environmental monitoring: An integrated model supporting enzyme activity and histopathological lesions

    Science.gov (United States)

    Neta, Raimunda Nonata Fortes Carvalho; Torres Junior, Audalio Rebelo

    2014-10-01

    We present a mathematical model describing the association between glutathione-S-transferase activity and brachial lesions in the catfish, Sciades herzbergii (Ariidae) from a polluted port. The catfish were sampled from a port known to be contaminated with heavy metals and organic compounds and from a natural reserve in São Marcos Bay, Brazil. Two biomarkers, hepatic glutathione S-transferase (GST) activity and histopathological lesions, in gills tissue were measured. The values for GST activity were modeled with the occurrence of branchial lesions by fitting a third order polynomial. Results from the mathematical model indicate that GST activity has a strong polynomial relationship with the occurrence of branchial lesions in both the wet and the dry seasons, but only at the polluted port site. The model developed in this study indicates that branchial and hepatic lesions are initiated when GST activity reaches 2.15 μmol min-1 mg protein-1. Beyond this limit, GST activity decreased to very low levels and irreversible histopathological lesions occurred. This mathematical model provides a realistic approach to analyze predictive biomarkers of environmental health status.

  15. Comparison of Nanostring nCounter® Data on FFPE Colon Cancer Samples and Affymetrix Microarray Data on Matched Frozen Tissues.

    Science.gov (United States)

    Chen, Xi; Deane, Natasha G; Lewis, Keeli B; Li, Jiang; Zhu, Jing; Washington, M Kay; Beauchamp, R Daniel

    2016-01-01

    The prognosis of colorectal cancer (CRC) stage II and III patients remains a challenge due to the difficulties of finding robust biomarkers suitable for testing clinical samples. The majority of published gene signatures of CRC have been generated on fresh frozen colorectal tissues. Because collection of frozen tissue is not practical for routine surgical pathology practice, a clinical test that improves prognostic capabilities beyond standard pathological staging of colon cancer will need to be designed for formalin-fixed paraffin-embedded (FFPE) tissues. The NanoString nCounter® platform is a gene expression analysis tool developed for use with FFPE-derived samples. We designed a custom nCounter® codeset based on elements from multiple published fresh frozen tissue microarray-based prognostic gene signatures for colon cancer, and we used this platform to systematically compare gene expression data from FFPE with matched microarray array data from frozen tissues. Our results show moderate correlation of gene expression between two platforms and discovery of a small subset of genes as candidate biomarkers for colon cancer prognosis that are detectable and quantifiable in FFPE tissue sections.

  16. Comparison of Nanostring nCounter® Data on FFPE Colon Cancer Samples and Affymetrix Microarray Data on Matched Frozen Tissues.

    Directory of Open Access Journals (Sweden)

    Xi Chen

    Full Text Available The prognosis of colorectal cancer (CRC stage II and III patients remains a challenge due to the difficulties of finding robust biomarkers suitable for testing clinical samples. The majority of published gene signatures of CRC have been generated on fresh frozen colorectal tissues. Because collection of frozen tissue is not practical for routine surgical pathology practice, a clinical test that improves prognostic capabilities beyond standard pathological staging of colon cancer will need to be designed for formalin-fixed paraffin-embedded (FFPE tissues. The NanoString nCounter® platform is a gene expression analysis tool developed for use with FFPE-derived samples. We designed a custom nCounter® codeset based on elements from multiple published fresh frozen tissue microarray-based prognostic gene signatures for colon cancer, and we used this platform to systematically compare gene expression data from FFPE with matched microarray array data from frozen tissues. Our results show moderate correlation of gene expression between two platforms and discovery of a small subset of genes as candidate biomarkers for colon cancer prognosis that are detectable and quantifiable in FFPE tissue sections.

  17. 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

  18. A Comparison of RNA-Seq Results from Paired Formalin-Fixed Paraffin-Embedded and Fresh-Frozen Glioblastoma Tissue Samples

    Science.gov (United States)

    Esteve-Codina, Anna; Arpi, Oriol; Martinez-García, Maria; Pineda, Estela; Mallo, Mar; Gut, Marta; Carrato, Cristina; Rovira, Anna; Lopez, Raquel; Tortosa, Avelina; Dabad, Marc; Del Barco, Sonia; Heath, Simon; Bagué, Silvia; Ribalta, Teresa; Alameda, Francesc; de la Iglesia, Nuria

    2017-01-01

    The molecular classification of glioblastoma (GBM) based on gene expression might better explain outcome and response to treatment than clinical factors. Whole transcriptome sequencing using next-generation sequencing platforms is rapidly becoming accepted as a tool for measuring gene expression for both research and clinical use. Fresh frozen (FF) tissue specimens of GBM are difficult to obtain since tumor tissue obtained at surgery is often scarce and necrotic and diagnosis is prioritized over freezing. After diagnosis, leftover tissue is usually stored as formalin-fixed paraffin-embedded (FFPE) tissue. However, RNA from FFPE tissues is usually degraded, which could hamper gene expression analysis. We compared RNA-Seq data obtained from matched pairs of FF and FFPE GBM specimens. Only three FFPE out of eleven FFPE-FF matched samples yielded informative results. Several quality-control measurements showed that RNA from FFPE samples was highly degraded but maintained transcriptomic similarities to RNA from FF samples. Certain issues regarding mutation analysis and subtype prediction were detected. Nevertheless, our results suggest that RNA-Seq of FFPE GBM specimens provides reliable gene expression data that can be used in molecular studies of GBM if the RNA is sufficiently preserved. PMID:28122052

  19. MMAD: microarray microdissection with analysis of differences is a computational tool for deconvoluting cell type-specific contributions from tissue samples.

    Science.gov (United States)

    Liebner, David A; Huang, Kun; Parvin, Jeffrey D

    2014-03-01

    One of the significant obstacles in the development of clinically relevant microarray-derived biomarkers and classifiers is tissue heterogeneity. Physical cell separation techniques, such as cell sorting and laser-capture microdissection, can enrich samples for cell types of interest, but are costly, labor intensive and can limit investigation of important interactions between different cell types. We developed a new computational approach, called microarray microdissection with analysis of differences (MMAD), which performs microdissection in silico. Notably, MMAD (i) allows for simultaneous estimation of cell fractions and gene expression profiles of contributing cell types, (ii) adjusts for microarray normalization bias, (iii) uses the corrected Akaike information criterion during model optimization to minimize overfitting and (iv) provides mechanisms for comparing gene expression and cell fractions between samples in different classes. Computational microdissection of simulated and experimental tissue mixture datasets showed tight correlations between predicted and measured gene expression of pure tissues as well as tight correlations between reported and estimated cell fraction for each of the individual cell types. In simulation studies, MMAD showed superior ability to detect differentially expressed genes in mixed tissue samples when compared with standard metrics, including both significance analysis of microarrays and cell type-specific significance analysis of microarrays. We have developed a new computational tool called MMAD, which is capable of performing robust tissue microdissection in silico, and which can improve the detection of differentially expressed genes. MMAD software as implemented in MATLAB is publically available for download at http://sourceforge.net/projects/mmad/.

  20. Histopathological comparison of bone healing effects of endonasal and percutaneous lateral osteotomy methods in rabbit rhinoplasty model.

    Science.gov (United States)

    Öğreden, Şahin; Rüzgar, Sedat; Tansuker, Hasan Deniz; Taşkın, Ümit; Alimoğlu, Yalçın; Aydın, Salih; Oktay, Mehmet Faruk; İzol, Uğur

    2017-07-17

    Lateral osteotomy is mainly performed either endonasally or percutaneously in rhinoplasty which is a frequently performed operation for the correction of nasal deformities. Both techniques have both advantages and disadvantages relative to each other. The aim of this study was to compare the histopathological effects of endonasal and percutaneous osteotomy techniques performed in rhinoplasty on bone healing and nasal stability in an experimental animal model. Eight one year-old New Zealand white rabbits were included. Xylazine hydrocloride and intramuscular ketamine anesthesia were administered to the rabbits. Endonasal osteotomy (8 bones) was performed in Group 1 (n=4), and percutaneous osteotomy (8 bones) in Group 2 (n=4). One month later the rabbits were sacrificed. Bone healing of the rabbits was staged according to the bone healing score of Huddleston et al. In both groups, nasal bone integrity was assessed subjectively. In the percutaneous osteotomy group, Grade 1 bone healing was observed in two samples (25%), Grade 2 bone healing in two samples (25%), Grade 3 bone healing in four samples (50%). In the endonasal osteotomy group, Grade 1 bone healing was observed in 6 samples (75%) and Grade 2 bone healing was observed in 2 samples (25%). In the percutaneous group, fibrous tissue was observed in 2, predominantly fibrous tissue and a lesser amount of cartilage was observed in 2 and an equal amount of fibrous tissue and cartilage was observed in 4 samples. In the endonasal group, fibrous tissue was observed in 6 samples, and predominantly fibrous tissue with a lesser amount of cartilage was observed in 2 samples. In both groups, when manual force was applied to the nasal bones, subjectively the same resistance was observed. Percutaneous lateral osteotomy technique was found to result in less bone and periost trauma and better bone healing compared to the endonasal osteotomy technique. Copyright © 2017 Associação Brasileira de Otorrinolaringologia e Cirurgia

  1. Occurrence of fruiting structures allows determination of Purpureocillium lilacinum as an inciting agent of pleuritis and pneumonia in a loggerhead sea turtle (Caretta caretta by histopathologic correlation to culture

    Directory of Open Access Journals (Sweden)

    V.L. Schumacher

    2014-10-01

    Full Text Available Purpureocillium lilacinum and Beauveria bassiana were isolated from lung sampled at necropsy of a 12 year-old female loggerhead sea turtle (Caretta caretta that had displayed abnormal buoyancy. Histopathologic evaluation revealed pleuritis and pneumonia with non-melanized, septate hyphae and fruiting structures identical to those of P. lilacinum. This case emphasizes the importance of a histological correlate to fungal culture when environmental fungi are isolated and demonstrates the infrequent phenomenon of fruiting or conidial production in tissue.

  2. Automated column liquid chromatographic determination of amoxicillin and cefadroxil in bovine serum and muscle tissue using on-line dialysis for sample preparation

    NARCIS (Netherlands)

    Snippe, N; van de Merbel, N C; Ruiter, F P; Steijger, O M; Lingeman, H; Brinkman, U A

    1994-01-01

    A fully automated method is described for the determination of amoxicillin and cefadroxil in bovine serum and muscle tissue. The method is based on the on-line combination of dialysis and solid-phase extraction for sample preparation, and column liquid chromatography with ultraviolet detection. In o

  3. Use of alkaline or enzymatic sample pretreatment prior to characterization of gold nanoparticles in animal tissue by single-particle ICPMS.

    Science.gov (United States)

    Loeschner, Katrin; Brabrand, Myung Suk Jung; Sloth, Jens J; Larsen, Erik H

    2014-06-01

    Inductively coupled plasma mass spectrometry in single-particle mode (spICPMS) is a promising method for the detection of metal-containing nanoparticles (NPs) and the quantification of their size and number concentration. Whereas existing studies mainly focus on NPs suspended in aqueous matrices, not much is known about the applicability of spICPMS for determination of NPs in complex matrices such as biological tissues. In the present study, alkaline and enzymatic treatments were applied to solubilize spleen samples from rats, which had been administered 60-nm gold nanoparticles (AuNPs) intravenously. The results showed that similar size distributions of AuNPs were obtained independent of the sample preparation method used. Furthermore, the quantitative results for AuNP mass concentration obtained with spICPMS following alkaline sample pretreatment coincided with results for total gold concentration obtained by conventional ICPMS analysis of acid-digested tissue. The recovery of AuNPs from enzymatically digested tissue, however, was approximately four times lower. Spiking experiments of blank spleen samples with AuNPs showed that the lower recovery was caused by an inferior transport efficiency of AuNPs in the presence of enzymatically digested tissue residues.

  4. Color Tissue Doppler to Analyze Fetal Cardiac Time Intervals: Normal Values and Influence of Sample Gate Size.

    Science.gov (United States)

    Willruth, A M; Steinhard, J; Enzensberger, C; Axt-Fliedner, R; Gembruch, U; Doelle, A; Dimitriou, I; Fimmers, R; Bahlmann, F

    2016-02-04

    Purpose: To assess the time intervals of the cardiac cycle in healthy fetuses in the second and third trimester using color tissue Doppler imaging (cTDI) and to evaluate the influence of different sizes of sample gates on time interval values. Materials and Methods: Time intervals were measured from the cTDI-derived Doppler waveform using a small and large region of interest (ROI) in healthy fetuses. Results: 40 fetuses were included. The median gestational age at examination was 26 + 1 (range: 20 + 5 - 34 + 5) weeks. The median frame rate was 116/s (100 - 161/s) and the median heart rate 143 (range: 125 - 158) beats per minute (bpm). Using small and large ROIs, the second trimester right ventricular (RV) mean isovolumetric contraction times (ICTs) were 39.8 and 41.4 ms (p = 0.17), the mean ejection times (ETs) were 170.2 and 164.6 ms (p < 0.001), the mean isovolumetric relaxation times (IRTs) were 52.8 and 55.3 ms (p = 0.08), respectively. The left ventricular (LV) mean ICTs were 36.2 and 39.4 ms (p = 0.05), the mean ETs were 167.4 and 164.5 ms (p = 0.013), the mean IRTs were 53.9 and 57.1 ms (p = 0.05), respectively. The third trimester RV mean ICTs were 50.7 and 50.4 ms (p = 0.75), the mean ETs were 172.3 and 181.4 ms (p = 0.49), the mean IRTs were 50.2 and 54.6 ms (p = 0.03); the LV mean ICTs were 45.1 and 46.2 ms (p = 0.35), the mean ETs were 175.2 vs. 172.9 ms (p = 0.29), the mean IRTs were 47.1 and 50.0 ms (p = 0.01), respectively. Conclusion: Isovolumetric time intervals can be analyzed precisely and relatively independent of ROI size. In the near future, automatic time interval measurement using ultrasound systems will be feasible and the analysis of fetal myocardial function can become part of the clinical routine.

  5. Neutrophil extracellular traps in tissue pathology.

    Science.gov (United States)

    Nakazawa, Daigo; Kumar, Santosh; Desai, Jyaysi; Anders, Hans-Joachim

    2017-03-01

    Neutrophil extracellular traps (NETs) are innate immune systems against invading pathogens. NETs are characterized as released DNA mixed with cytoplasmic antimicrobial proteins such as myeloperoxidase, proteinase3 and neutrophil elastase. While NETs are thought to have an important role in host defense, recent work has suggested that NETs contribute to tissue injury in non-infectious disease states. Uncontrolled NET formation in autoimmune diseases, metabolic disorders, cancers and thrombotic diseases can exacerbate a disease or even be a major initiator of tissue injury. But spotting NETs in tissues is not easy. Here we review the available histopathological evidence on the presence of NETs in a variety of diseases. We discuss technical difficulties and potential sources of misinterpretation while trying to detect NETs in tissue samples.

  6. Nonlesions, misdiagnoses, missed diagnoses, and other interpretive challenges in fish histopathology studies: a guide for investigators, authors, reviewers, and readers

    Science.gov (United States)

    Wolf, Jeffrey C.; Baumgartner, Wes A.; Blazer, Vicki; Camus, Alvin C.; Engelhardt, Jeffrey A.; Fournie, John W.; Frasca, Salvatore; Groman, David B.; Kent, Michael L.; Khoo, Lester H.; Law, Jerry M.; Lombardini, Eric D.; Ruehl-Fehlert, Christine; Segner, Helmut E.; Smith, Stephen A.; Spitsbergen, Jan M.; Weber, Klaus; Wolfe, Marilyn J.

    2015-01-01

    Differentiating salient histopathologic changes from normal anatomic features or tissue artifacts can be decidedly challenging, especially for the novice fish pathologist. As a consequence, findings of questionable accuracy may be reported inadvertently, and the potential negative impacts of publishing inaccurate histopathologic interpretations are not always fully appreciated. The objectives of this article are to illustrate a number of specific morphologic findings in commonly examined fish tissues (e.g., gills, liver, kidney, and gonads) that are frequently either misdiagnosed or underdiagnosed, and to address related issues involving the interpretation of histopathologic data. To enhance the utility of this article as a guide, photomicrographs of normal and abnormal specimens are presented. General recommendations for generating and publishing results from histopathology studies are additionally provided. It is hoped that the furnished information will be a useful resource for manuscript generation, by helping authors, reviewers, and readers to critically assess fish histopathologic data.

  7. Nonlesions, misdiagnoses, missed diagnoses, and other interpretive challenges in fish histopathology studies: a guide for investigators, authors, reviewers, and readers.

    Science.gov (United States)

    Wolf, Jeffrey C; Baumgartner, Wes A; Blazer, Vicki S; Camus, Alvin C; Engelhardt, Jeffery A; Fournie, John W; Frasca, Salvatore; Groman, David B; Kent, Michael L; Khoo, Lester H; Law, Jerry M; Lombardini, Eric D; Ruehl-Fehlert, Christine; Segner, Helmut E; Smith, Stephen A; Spitsbergen, Jan M; Weber, Klaus; Wolfe, Marilyn J

    2015-04-01

    Differentiating salient histopathologic changes from normal anatomic features or tissue artifacts can be decidedly challenging, especially for the novice fish pathologist. As a consequence, findings of questionable accuracy may be reported inadvertently, and the potential negative impacts of publishing inaccurate histopathologic interpretations are not always fully appreciated. The objectives of this article are to illustrate a number of specific morphologic findings in commonly examined fish tissues (e.g., gills, liver, kidney, and gonads) that are frequently either misdiagnosed or underdiagnosed, and to address related issues involving the interpretation of histopathologic data. To enhance the utility of this article as a guide, photomicrographs of normal and abnormal specimens are presented. General recommendations for generating and publishing results from histopathology studies are additionally provided. It is hoped that the furnished information will be a useful resource for manuscript generation, by helping authors, reviewers, and readers to critically assess fish histopathologic data.

  8. Influence of parasite density and sample storage time on the reliability of Entamoeba histolytica-specific PCR from formalin-fixed and paraffin-embedded tissues.

    Science.gov (United States)

    Frickmann, Hagen; Tenner-Racz, Klara; Eggert, Petra; Schwarz, Norbert G; Poppert, Sven; Tannich, Egbert; Hagen, Ralf M

    2013-12-01

    We report on the reliability of polymerase chain reaction (PCR) for the detection of Entamoeba histolytica from formalin-fixed, paraffin-embedded tissue in comparison with microscopy and have determined predictors that may influence PCR results. E. histolytica-specific and Entamoeba dispar-specific real-time PCR and microscopy from adjacent histologic sections were performed using a collection of formalin-fixed, paraffin-embedded tissue specimens obtained from patients with invasive amebiasis. Specimens had been collected during the previous 4 decades. Association of sample age, parasite density, and reliability of PCR was analyzed. E. histolytica PCR was positive in 20 of 34 biopsies (58.8%); 2 of these 20 were microscopically negative for amebae in neighboring tissue sections. PCR was negative in 9 samples with visible amebae in neighboring sections and in 5 samples without visible parasites in neighboring sections. PCR was negative in all specimens that were older than 3 decades. Low parasite counts and sample ages older than 20 years were predictors for false-negative PCR results. All samples were negative for E. dispar DNA. PCR is suitable for the detection of E. histolytica in formalin-fixed, paraffin-embedded tissue samples that are younger than 2 decades and that contain intermediate to high parasite numbers. Negative results in older samples were due to progressive degradation of DNA over time as indicated by control PCRs targeting the human 18S rRNA gene. Moreover, our findings support previous suggestions that only E. histolytica but not E. dispar is responsible for invasive amebiasis.

  9. Histopathological Image Classification Using Discriminative Feature-Oriented Dictionary Learning.

    Science.gov (United States)

    Vu, Tiep Huu; Mousavi, Hojjat Seyed; Monga, Vishal; Rao, Ganesh; Rao, U K Arvind

    2016-03-01

    In histopathological image analysis, feature extraction for classification is a challenging task due to the diversity of histology features suitable for each problem as well as presence of rich geometrical structures. In this paper, we propose an automatic feature discovery framework via learning class-specific dictionaries and present a low-complexity method for classification and disease grading in histopathology. Essentially, our Discriminative Feature-oriented Dictionary Learning (DFDL) method learns class-specific dictionaries such that under a sparsity constraint, the learned dictionaries allow representing a new image sample parsimoniously via the dictionary corresponding to the class identity of the sample. At the same time, the dictionary is designed to be poorly capable of representing samples from other classes. Experiments on three challenging real-world image databases: 1) histopathological images of intraductal breast lesions, 2) mammalian kidney, lung and spleen images provided by the Animal Diagnostics Lab (ADL) at Pennsylvania State University, and 3) brain tumor images from The Cancer Genome Atlas (TCGA) database, reveal the merits of our proposal over state-of-the-art alternatives. Moreover, we demonstrate that DFDL exhibits a more graceful decay in classification accuracy against the number of training images which is highly desirable in practice where generous training is often not available.

  10. Vitamin B12 deficiency and gastric histopathology in older patients

    Institute of Scientific and Technical Information of China (English)

    KR Dholakia; TS Dharmarajan; D Yadav; S Oiseth; EP Norkus; CS Pitchumoni

    2005-01-01

    AIM: To compare upper gastric endoscopic and histopathologic findings in older adults in the presence and absence of B12 deficiency.METHODS: A prospective analysis of upper gastric endoscopic and gastric histopathologic findings from 30 newly identified B12-deficient patients (11 males,19 females) and 16 controls with normal B12 status (6males, 10 females) was performed. For all subjects, the indication for upper endoscopy and gastric biopsy were unrelated to B12 status. A single pathologist, blinded to B12 status, processed and interpreted the biopsy samples. Endoscopic and histopathologic findings were correlated with age, gender, hematocrit (Hct), MCV and B12 status.RESULTS: The B12-deficient group had significantly lower mean serum B12 levels compared to the controls (P<0.00005) while their mean Hct, MCV and serum albumin levels were similar. Iron deficiency (ferritinbased) was present in 21% of B12-deficient patients and intrinsic factor antibodies were present in29% (5/17) of B12-deficient patients. The endoscopic findings revealed significantly different rates of gastritis and atrophy between the B12-deficient and control groups (P= 0.017).B12-deficient patients had significantly less superficial gastritis (62% vs 94%) and significantly more atrophic gastritis (28% vs 0%) as compared to the controls (P= 0.039). Intestinal metaplasia was similar in both groups. Helicobacter pyloriinfection rates were similar in the B12-deficient patients and controls (40% vs31%).CONCLUSION: Significantly different endoscopic findings and types of gastritis could often be observed in the presence and absence of B12 deficiency. Atrophy,based on endoscopy, and atrophic gastritis, based on histopathology, suggest the presence of B12 deficiency.Gastric histopathology is not influenced by the age,gender, Hct or MCV of the patients.

  11. Some Physical, Chemical, and Biological Parameters of Samples of Scleractinium Coral Aquaculture Skeleton Used for Reconstruction/Engineering of the Bone Tissue.

    Science.gov (United States)

    Popov, A A; Sergeeva, N S; Britaev, T A; Komlev, V S; Sviridova, I K; Kirsanova, V A; Akhmedova, S A; Dgebuadze, P Yu; Teterina, A Yu; Kuvshinova, E A; Schanskii, Ya D

    2015-08-01

    Physical and chemical (phase and chemical composition, dynamics of resorption, and strength properties), and biological (cytological compatibility and scaffold properties of the surface) properties of samples of scleractinium coral skeletons from aquacultures of three types and corresponding samples of natural coral skeletons (Pocillopora verrucosa, Acropora formosa, and Acropora nobilis) were studied. Samples of scleractinium coral aquaculture skeleton of A. nobilis, A. formosa, and P. verrucosa met the requirements (all study parameters) to materials for osteoplasty and 3D-scaffolds for engineering of bone tissue.

  12. Intraoperative tissue identification using rapid evaporative ionization mass spectrometry.

    Science.gov (United States)

    Balog, Júlia; Sasi-Szabó, László; Kinross, James; Lewis, Matthew R; Muirhead, Laura J; Veselkov, Kirill; Mirnezami, Reza; Dezső, Balázs; Damjanovich, László; Darzi, Ara; Nicholson, Jeremy K; Takáts, Zoltán

    2013-07-17

    Rapid evaporative ionization mass spectrometry (REIMS) is an emerging technique that allows near-real-time characterization of human tissue in vivo by analysis of the aerosol ("smoke") released during electrosurgical dissection. The coupling of REIMS technology with electrosurgery for tissue diagnostics is known as the intelligent knife (iKnife). This study aimed to validate the technique by applying it to the analysis of fresh human tissue samples ex vivo and to demonstrate the translation to real-time use in vivo in a surgical environment. A variety of tissue samples from 302 patients were analyzed in the laboratory, resulting in 1624 cancerous and 1309 noncancerous database entries. The technology was then transferred to the operating theater, where the device was coupled to existing electrosurgical equipment to collect data during a total of 81 resections. Mass spectrometric data were analyzed using multivariate statistical methods, including principal components analysis (PCA) and linear discriminant analysis (LDA), and a spectral identification algorithm using a similar approach was implemented. The REIMS approach differentiated accurately between distinct histological and histopathological tissue types, with malignant tissues yielding chemical characteristics specific to their histopathological subtypes. Tissue identification via intraoperative REIMS matched the postoperative histological diagnosis in 100% (all 81) of the cases studied. The mass spectra reflected lipidomic profiles that varied between distinct histological tumor types and also between primary and metastatic tumors. Thus, in addition to real-time diagnostic information, the spectra provided additional information on divergent tumor biochemistry that may have mechanistic importance in cancer.

  13. Retrieval and Amplification of DNA from Unstained Histopathological Sections

    Institute of Scientific and Technical Information of China (English)

    DonnaC.MONTAGUE; BeverlyD.LYN-COOK; 等

    1993-01-01

    Testing of compounds for carcinogenic potential in vivo involves various experimental designs.A few of these techniques are directed to demonstrate the genotoxicity and mutagenicity of the compound by histopathology.These changes shown by histochemical means include monoclonal antibody directed cellular markers.Development of the polymerase chain reaction technique(PCR)for amplification of DNA has facilitated the investigation of molecular events related to the formation of malignant neoplasms.We describe here a method for screening tissues for mutations of the H-ras gene using monoclonal antibodies directed toward normal and mutant p21 proteins.Formalin-fixed,paraffinembedded tissue sections are used to subsequently confirm the gene mutation by PCR amplification of the H-ras gene.The results indicated a successful application of this technique to demonstrate the presence of p21 oncoprotein in the tissues tested.

  14. Stain-free histopathology by programmable supercontinuum pulses

    Science.gov (United States)

    Tu, Haohua; Liu, Yuan; Turchinovich, Dmitry; Marjanovic, Marina; Lyngsø, Jens K.; Lægsgaard, Jesper; Chaney, Eric J.; Zhao, Youbo; You, Sixian; Wilson, William L.; Xu, Bingwei; Dantus, Marcos; Boppart, Stephen A.

    2016-08-01

    The preparation, staining, visualization and interpretation of histological images of tissue is well accepted as the gold standard process for the diagnosis of disease. These methods have a long history of development, and are used ubiquitously in pathology, despite being highly time- and labour-intensive. Here, we introduce a unique optical imaging platform and methodology for label-free multimodal multiphoton microscopy that uses a novel photonic-crystal fibre source to generate tailored chemical contrast based on programmable supercontinuum pulses. We demonstrate the collection of optical signatures of the tumour microenvironment, including evidence of mesoscopic biological organization, tumour cell migration and (lymph-) angiogenesis collected directly from fresh ex vivo mammary tissue. Acquisition of these optical signatures and other cellular or extracellular features, which are largely absent from histologically processed and stained tissue, combined with an adaptable platform for optical alignment-free programmable-contrast imaging, offers the potential to translate stain-free molecular histopathology into routine clinical use.

  15. Suitability of faeces and tissue samples as a basis for non-invasive sampling for African swine fever in wild boar

    NARCIS (Netherlands)

    Carvalho Ferreira, de H.C.; Weesendorp, E.; Quak, S.; Stegeman, J.A.; Loeffen, W.L.A.

    2014-01-01

    A challenging aspect of ASFV control in wild boar populations is the design and implementation of effective surveillance and monitoring programmes, both for early warning, and to determine the ongoing epidemiological situation in an infected population. Testing blood samples requires invasive sampli

  16. The effect of co-occurring polychlorinated biphenyls on quantitation of toxaphene in fish tissue samples by gas chromatography negative ion mass spectrometry.

    Science.gov (United States)

    Lao, Wenjian; Tsukada, David; Maruya, Keith A

    2012-12-28

    Determinative methods based on gas chromatography-negative chemical ionization mass spectrometry (GC-NCI/MS) provide improved sensitivity and specificity for toxaphene in environmental samples, but are subject to misidentification due to oxygen reaction in the presence of polychlorinated biphenyls (PCBs). The goal of this study was to quantify the impact of co-occurring PCBs in fish tissue samples when utilizing single quadrupole instruments to implement this method. Mixtures of PCB congeners and technical toxaphene, and extracts of fish tissue with varying concentrations of PCBs were analyzed for individual congener and total toxaphene concentrations by GC-NCI/MS. The contribution of co-injected PCB 204 ranged from 23% to 88% of the total peak area for the Cl-9 toxaphene homolog quantitation ion, a contribution that increased as the ratio of technical toxaphene to PCB 204 decreased. PCB interferences in fish tissue extracts, including a standard reference material, were subtracted using a three-step procedure featuring spectral analysis of isotopic patterns for target peaks. Total toxaphene concentrations without PCB subtraction in three fish tissue samples with low, intermediate and high co-occurring PCBs were overestimated by 33, 55 and 745%, respectively, underscoring the need for practical strategies to account for PCB interferences in GC-NCI/MS based protocols. In contrast, no appreciable interference or resulting positive bias in concentrations was observed for quantitation of eight common toxaphene residue congeners.

  17. Fibroadenomas of the breast showing an ill-defined margin on ultrasonogram: Correlation with histopathologic findings

    Energy Technology Data Exchange (ETDEWEB)

    Hwang, Ji Young; Choi, Hye Young; Shim, Sung Shine; Rhee, Chung Sik; Sung, Soon Hee [Ewha Womans University College of Medicine, Seoul (Korea, Republic of)

    2002-12-15

    To correlate the sonographic finding of ill-defined fibroadenoma with the histopathologic findings. Sonographic finding of forty nine surgically proven fibroadenomas were retrospectively correlated with histopathologic findings. We evaluated sonographic findings including the margin, shape, size and the echotexture of fibroadenoma. The histopathologic glandular structure, stromal type of fibroadenoma, type of the surrounding breast tissue and presence of interdigitation between fibroadenoma and the surrounding breast tissue were pathologically reviewed and analyzed statistically. Breast sonography of fibroadenomas demonstrated well-defined margin in 28 (57%) and ill-defined margin in 21 (43%) among 49 lesions. Histopathologically, the glandular structure and type of the surrounding breast tissue were not significantly different between well-defined and ill-defined fibroadenomas. The stromal type of fibroadenoma was sclerotic in 8 (29%) well-defined fibroadenomas while sclerotic in 15 (72%) of 21 ill-defined fibroadenomas,showing difference with a statistical significance between well-defined and ill-defined fibroadenomas (p<0.05). Twenty (71%) of 28 fibroadenomas with well-defined sonographic margins showed well-defined border between mass and the surrounding breast tissue, pathologically. Meanwhile, twenty (95%) of 21 fibroadenomas with ill-defined sonographic margins demonstrated interdigitation of the surrounding breast tissue and mass, exhibiting a statistical significance (p<0.005). Ill-defined margins of fibroadenomas on sonography represent the interdigitation of the surrounding breast tissue with a mass and is seen in the sclerotic stromal type of fibroadenomas.

  18. Conjunctival tumors in children: histopathologic diagnosis in 165 cases

    Directory of Open Access Journals (Sweden)

    Martin A. Zimmermann-Paiz

    2015-12-01

    Full Text Available ABSTRACT Purpose: Conjunctival tissue tumors have a varied presentation, and few series studies on pediatric patients have been published. The objective of this paper is to report the histopathologic diagnoses (spanning over 1988-2013 of conjunctival tumors in children younger than 14 years. Methods: We conducted a retrospective, descriptive, and observational study by reviewing the database of all children in whom a conjunctival tumor was surgically removed at Hospital de Ojos y Oídos "Dr. Rodolfo Robles V.," Benemérito Comité Pro Ciegos y Sordos de Guatemala. The data pertaining to gender, age, and histopathologic diagnosis of all cases was collected. The same ocular pathologist made all diagnoses. Results: One hundred sixty-five cases were found, with a mean age of 7.88 years, being 91 (55.15% male subjects. Melanocytic lesions were the most common tumors found (30.91% of cases, with only one case (0.60% being malignant. Conclusions: Melanocytic lesions were the most common tumors found, and of all the cases, only one was malignant; this was in a patient with xeroderma pigmentosum. These findings are consistent with those reported in other studies regarding the frequencies of the histopathology of conjunctival tumors in the pediatric population.

  19. Determination of boron concentration in blood and tissue samples from patients with liver metastases of colorectal carcinoma using Prompt Gamma Ray Activation Analysis (PGAA)

    Energy Technology Data Exchange (ETDEWEB)

    Schmitz, T., E-mail: schmito@uni-mainz.d [Institute for Nuclear Chemistry, University of Mainz, Fritz-Strassmann-Weg 2, D-55128 Mainz (Germany); Appelman, K., E-mail: k.appelman@hetnet.n [Institute for Energy, Joint Research Centre of the European Commission, Petten (Netherlands); Kudejova, P., E-mail: petra.kudejova@frm2.tum.d [Forschungs-Neutronenquelle Heinz Maier-Leibnitz (FRM II), Technische Universitaet Muenchen, D-85748 Garching (Germany); Schuetz, C., E-mail: schuetc@uni-mainz.d [Institute for Nuclear Chemistry, University of Mainz, Fritz-Strassmann-Weg 2, D-55128 Mainz (Germany); Kratz, J.V., E-mail: jvkratz@uni-mainz.d [Institute for Nuclear Chemistry, University of Mainz, Fritz-Strassmann-Weg 2, D-55128 Mainz (Germany); Moss, R., E-mail: raymond.moss@ec.europa.e [Institute for Energy, Joint Research Centre of the European Commission, Petten (Netherlands); Otto, G., E-mail: gerd.otto@unimedizin-mainz.d [Department of Hepatobiliary, Pancreatic and Transplantation Surgery, University of Mainz, Langenbeckstr. 1, D-55131 Mainz (Germany); Hampel, G., E-mail: gabriele.hampel@uni-mainz.d [Institute for Nuclear Chemistry, University of Mainz, Fritz-Strassmann-Weg 2, D-55128 Mainz (Germany)

    2011-07-15

    As part of the studies on Boron Neutron Capture Therapy at the University of Mainz, Germany, a clinical trial has been started in which, four patients suffering from liver metastases of colorectal carcinoma have been enrolled. Specimens of blood and healthy tissue samples taken from the patients were measured at the PGAA facilities at the HFR in Petten, The Netherlands, and at the FRM II in Munich, Germany. From the measured boron concentrations, pharmacokinetic curves and blood-to-tissue concentration ratios were produced.

  20. Study on outbreak ofNeospora caninum-associated abortion in dairy cows in Tabriz (Northwest Iran) by serological, molecular and histopathologic methods

    Institute of Scientific and Technical Information of China (English)

    A Nematollahi; G H Moghaddam; R Jaafari; J Ashrafi Helan; M Norouzi

    2013-01-01

    Objective:To determineNeospora caninum(N. caninum) as a cause of bovine abortion in dairy cows byELISA,PCR andPathological methods inTabriz,Northwest ofIran.Methods:For study of outbreak of neosporosis, blood samples were collected from76Holstein aborted dairy cows in Tabriz(NorthwestIran).Antibodies to N. caninum were assayed by using a commerciallyELISA kit(IDEXX,USA).IgG againstN. caninum were found in14(18.4%) cases.Aborted fetuses of these seropositive dams were proposed for histopathological and molecular investigations.Brains, spinal cords and placentas of the fetuses were fixed in10% buffered formalin for histopathology. Also5-10 g of brain tissue was sampled forDNA extraction.In6 out of14(42.8%) fetuses, brain tissue was positive inPCR.All dams of these fetuses were serologically seropositive inELISA test. Results:Histopathologically, the lesions consistent withN. caninumwere observed in brains, spinal cords and placenta of all fetuses.Lesions inCNS included severe congestion, perivascular and perineuronal edema, status spongiosis, perivascular cuffing, focal gliosis, nourophagy and focal necrosis.There were someNeospora-like cysts in brain.In placentas, severe congestion, perivascular infiltration of mononuclear cells, vascular thrombosis, focal placentitis and necrotic foci in cotyledons were noticed.Conclusions:The results of present study agreed the results of similar studies about serological, histopathological and molecular results of other studies about neosporosis and it seems to support the outbreak ofN. caninum-associated abortion in dairy cows inTabriz(NorthwestIran).

  1. Elastoma: clinical and histopathological aspects of a rare disease*

    Science.gov (United States)

    Maciel, Marina Gagheggi; Enokihara, Milvia Maria Simões e Silva; Seize, Maria Bandeira de Melo Paiva; Marcassi, Aline Pantano; Piazza, Christiane Affonso De Donato; Cestari, Silmara da Costa Pereira

    2016-01-01

    Elastoma is a connective tissue nevus characterized by changes in elastic fibers. It can be congenital or acquired, and is usually diagnosed before puberty. Associated with osteopoikilosis, it is known as Buschke-Ollendorff syndrome. Histopathology with specific staining for elastic fibers is critical for a diagnostic conclusion. This report describes the case of a 7-year-old male patient with lesions diagnosed as elastoma, with absence of bone changes in the radiological imaging. This study aims to report the clinical presentation and histological examination of such unusual disease.

  2. Vitamin-K-Dependent Protection of the Renal Microvasculature: Histopathological Studies in Normal and Diseased Kidneys

    Science.gov (United States)

    Wei, Fang-Fei; Drummen, Nadja E.A.; Thijs, Lutgarde; Jacobs, Lotte; Herfs, Marjolein; van't Hoofd, Cynthia; Vermeer, Cees; Staessen, Jan A.

    2016-01-01

    Vitamin-K-dependent carboxylation of matrix Gla protein (MGP) protects the macrocirculation against calcification. We recently reported in a multiethnic population study that the estimated glomerular filtration rate, a microvascular trait, decreased and the risk of chronic kidney disease increased with higher circulating levels of inactive dephospho-uncarboxylated MGP, a marker of vitamin K deficiency. These findings highlighted the possibility that vitamin K might have a beneficial effect on the renal microcirculation. To substantiate these epidemiological findings, we undertook a pilot study, in which we stained renal tissue samples obtained by biopsy from 2 healthy kidney donors and 4 patients with nephropathy for carboxylated and uncarboxylated MGP and calcium deposits. Three patients had renal calcifications, which were consistently associated with carboxylated and uncarboxylated MGP. Normal renal tissue was devoid of microcalcifications and staining for carboxylated and uncarboxylated MGP. Pending confirmation in a larger study covering a wider range of renal pathologies, these histopathological findings suggest that MGP might inhibit calcification not only in large arteries, as was known before, but in renal tissue as well, thereby highlighting potentially new avenues for promoting renal health, for instance by vitamin K supplementation.

  3. Histopathologic changes during mesenteric ischaemia and ...

    African Journals Online (AJOL)

    administration of drugs (ketamine, heparin) and for administra- tion of saline given to ... Table 1 Histopathologic scoring based on я modification of the Swerdlow .... cal and contractile activities of the canine small intestines. Am. J. Surg. 1976 ...

  4. Assessment of organosomatic indices and histopathological ...

    African Journals Online (AJOL)

    Assessment of organosomatic indices and histopathological changes in vital organs of ... of fish fed 0 and 20% BSSM inclusions showed no visible signs of damage. ... 20% in C. gariepinus' diet could cause adverse histological alterations.

  5. Breast Hemangioma: MR Appearance with Histopathological Correlation

    Directory of Open Access Journals (Sweden)

    Rasha Ameen

    2012-01-01

    Full Text Available Breast hemangioma is a rare tumor and when small, it may be difficult to diagnose using conventional imaging techniques. In this report the MR appearance is described with histopathological correlation.

  6. Renal and hepatic histopathology of intraperitoneally administered ...

    African Journals Online (AJOL)

    user

    2016-10-12

    Oct 12, 2016 ... The biochemistry and histopathology of intraperitoneally administered potassium ... and necrosis of crops in birds. ..... The stressful behavior of respiratory impairment .... specific bile acid transport system) which is responsible.

  7. The magnetization transfer characteristics of human breast tissues: an in vitro NMR study

    Science.gov (United States)

    Callicott, C.; Thomas, J. M.; Goode, A. W.

    1999-05-01

    A series of freshly excised human breast tissues was analysed using a nuclear magnetic resonance spectrometer and then subjected to routine histopathology examination. Tissues comprised normal parenchymal, adipose, fibrocystic, fibroadenoma and malignant types. An inversion-recovery sequence performed both with and without magnetization transfer allowed T1, T1, and values to be obtained. From this information, the magnetization transfer rate constant, K, was calculated for each tissue sample. These data show that T1 provided greater discrimination between neoplasic and normal tissues than did T1. However, neither T1 nor K values provided a means of discriminating between benign and malignant disease.

  8. Two alternative DNA extraction methods to improve the detection of Mycobacterium-tuberculosis-complex members in cattle and red deer tissue samples.

    Science.gov (United States)

    Fell, Shari; Bröckl, Stephanie; Büttner, Mathias; Rettinger, Anna; Zimmermann, Pia; Straubinger, Reinhard K

    2016-09-15

    Bovine tuberculosis (bTB), which is caused by Mycobacterium bovis and M. caprae, is a notifiable animal disease in Germany. Diagnostic procedure is based on a prescribed protocol that is published in the framework of German bTB legislation. In this protocol small sample volumes are used for DNA extraction followed by real-time PCR analyses. As mycobacteria tend to concentrate in granuloma and the infected tissue in early stages of infection does not necessarily show any visible lesions, it is likely that DNA extraction from only small tissue samples (20-40 mg) of a randomly chosen spot from the organ and following PCR testing may result in false negative results. In this study two DNA extraction methods were developed to process larger sample volumes to increase the detection sensitivity of mycobacterial DNA in animal tissue. The first extraction method is based on magnetic capture, in which specific capture oligonucleotides were utilized. These nucleotides are linked to magnetic particles and capture Mycobacterium-tuberculosis-complex (MTC) DNA released from 10 to 15 g of tissue material. In a second approach remaining sediments from the magnetic capture protocol were further processed with a less complex extraction protocol that can be used in daily routine diagnostics. A total number of 100 tissue samples from 34 cattle (n = 74) and 18 red deer (n = 26) were analyzed with the developed protocols and results were compared to the prescribed protocol. All three extraction methods yield reliable results by the real-time PCR analysis. The use of larger sample volume led to a sensitivity increase of DNA detection which was shown by the decrease of Ct-values. Furthermore five samples which were tested negative or questionable by the official extraction protocol were detected positive by real time PCR when the alternative extraction methods were used. By calculating the kappa index, the three extraction protocols resulted in a moderate (0.52; protocol 1 vs 3

  9. 应用子宫内膜细胞采集器获取微量子宫内膜组织行组织病理学诊断的临床分析%Clinical analysis of histopathology diagnosis of micro endometrial tissues obtained by endometrial cell collector

    Institute of Scientific and Technical Information of China (English)

    任玲; 王海燕; 魏媛; 杨静; 周健

    2016-01-01

    Objective To evaluate the feasibility and application of histopathology diagnosis of endometrial tissues obtained by endometrial cell collector (Jingyou, SAP-1). Methods One hundred and ninety-three patients whose endometrial lesions should be excluded were selected. First, endometrial tissue were obtained from the patients by Jingyou, then they underwent comprehensive curettage under hysteroscopy. The histopathology diagnosis was performed respectively. The specimen satisfaction rate and diagnostic accuracy was analyzed and compared. Results The specimen satisfaction rate of curettage under hysteroscopy was 95.85%(185/193). The specimens of 8 cases were not satisfied because the tissues were not enough. The specimen satisfaction rate of Jingyou was 82.38% (159/193). The specimens of 34 cases were not satisfied, among whom in 10 cases scratches did not throughout the whole palace antrum, and in 24 cases tissues quality were poor. The endometrial thickness in unsatisfactory specimen by Jinyou was significantly thinner than that in satisfactory specimen:(0.64 ± 0.18) cm vs. (0.97 ± 0.43) cm, and there was statistical difference (P<0.05). The diagnostic accordance rate between Jingyou and curettage under hysteroscopy was 79.87%(127/159). The sensitivity of Jingyou from high to low was 94.19% (81/86) in normal menstrual endometrial, 7/10 in endometrial carcinoma/ atypical hyperplasia, 67.86%(38/56) in endometrial hyperplasia and 1/7 in endometrial polyps. Missed diagnosis of jingyou inluded 2 cases of endometrial carcinoma and 4 cases of endometrial atypical hyperplasia. The misdiagnosed rate of high grade endometrial lesions was 6/16, and the patients were misdiagnosed because the tissues were not enough. Four cases of endometrial atypical hyperplasia had underwent conservative treatment of repeated curettage. Conclusions Application of Jingyou can obtain micro endometrial tissues, and the accordance rate of histopathology diagnosis is high with curettage under

  10. Microbiological and histopathological aspects of canine pyometra

    Science.gov (United States)

    Coggan, Jennifer Anne; Melville, Priscilla Anne; de Oliveira, Clair Motos; Faustino, Marcelo; Moreno, Andréa Micke; Benites, Nilson Roberti

    2008-01-01

    As pyometra is recognized as one of the main causes of disease and death in the bitch the purposes of this study were to evaluate microbiological and histopathological aspects of canine pyometra and to research the virulence factors of the E. coli isolates identifying possible risks to human health. The microbiological isolation from the intrauterine contents of 100 dogs with pyometra was carried out and the virulence factors in the E. coli strains were identified using PCR method. This study also consisted of the counting of microorganisms colonies forming units in samples of intrauterine content, tests of antimicrobial susceptibility of the E. coli isolates and the histological examination of the uterus. E. coli was the most prevalent microorganism isolated (76.6%) and 120 strains (79.5%) were positive for sfa, 86 (56.9%) were positive for cnf, 87 (57.6%) were positive for pap, 52 (34.4%) were positive for hly, 51 (33.8%) were positive for iuc and 5 (3.3%) were positive for afa genes. One observed more sensitivity of E. coli to norfloxacin, polimixin B, sulphazotrin, chloranfenicol and enrofloxacin. In 42% of the samples of uterine walls where microorganisms were isolated, the sizes of the areas of the inflammatory responses corresponded to 39–56%. Virulence factors were identified in 98.0% of the strains evaluated, demonstrating a high frequency of potentially pathogenic E. coli. It must be considered that dogs are animals that are living in close proximity to man for thousands of years and have an important role in the transmission of E. coli to other animals and to man. PMID:24031249

  11. Application of non-lethal stable isotope analysis to assess feeding patterns of juvenile pallid sturgeon Scaphirhynchus albus: a comparison of tissue types and sample preservation methods

    Science.gov (United States)

    Andvik, R.T.; VanDeHey, J.A.; Fincel, M.J.; French, William E.; Bertrand, K.N.; Chipps, Steven R.; Klumb, Robert A.; Graeb, B.D.S.

    2010-01-01

    Traditional techniques for stable isotope analysis (SIA) generally require sacrificing animals to collect tissue samples; this can be problematic when studying diets of endangered species such as the pallid sturgeon Scaphirhynchus albus. Our objectives were to (i) determine if pectoral fin tissue (non-lethal) could be a substitute for muscle tissue (lethal) in SIA of juvenile pallid sturgeon, and (ii) evaluate the influence of preservation techniques on stable isotope values. In the laboratory, individual juvenile pallid sturgeon were held for up to 186 day and fed chironomids, fish, or a commercially available pellet diet. Significant, positive relationships (r² ≥ 0.8) were observed between fin and muscle tissues for both δ15N and δ13C; in all samples isotopes were enriched in fins compared to muscle tissue. Chironomid and fish based diets of juvenile pallid sturgeon were distinguishable for fast growing fish (0.3 mm day−1) using stable δ15N and δ13C isotopes. Frozen and preserved fin tissue δ15N isotopes were strongly related (r2 = 0.89) but δ13C isotopes were weakly related (r2 = 0.16). Therefore, freezing is recommended for preservation of fin clips to avoid the confounding effect of enrichment by ethanol. This study demonstrates the utility of a non-lethal technique to assess time integrated food habits of juvenile pallid sturgeon and should be applicable to other threatened or endangered species.

  12. LINE-1 Hypomethylation in Blood and Tissue Samples as an Epigenetic Marker for Cancer Risk: A Systematic Review and Meta-Analysis

    Science.gov (United States)

    Barchitta, Martina; Quattrocchi, Annalisa; Maugeri, Andrea; Vinciguerra, Manlio; Agodi, Antonella

    2014-01-01

    Objective A systematic review and a meta-analysis were carried out in order to summarize the current published studies and to evaluate LINE-1 hypomethylation in blood and other tissues as an epigenetic marker for cancer risk. Methods A systematic literature search in the Medline database, using PubMed, was conducted for epidemiological studies, published before March 2014. The random-effects model was used to estimate weighted mean differences (MDs) with 95% Confidence Intervals (CIs). Furthermore, subgroup analyses were conducted by sample type (tissue or blood samples), cancer types, and by assays used to measure global DNA methylation levels. The Cochrane software package Review Manager 5.2 was used. Results A total of 19 unique articles on 6107 samples (2554 from cancer patients and 3553 control samples) were included in the meta-analysis. LINE-1 methylation levels were significantly lower in cancer patients than in controls (MD: −6.40, 95% CI: −7.71, −5.09; p<0.001). The significant difference in methylation levels was confirmed in tissue samples (MD −7.55; 95% CI: −9.14, −65.95; p<0.001), but not in blood samples (MD: −0.26, 95% CI: −0.69, 0.17; p = 0.23). LINE-1 methylation levels were significantly lower in colorectal and gastric cancer patients than in controls (MD: −8.33; 95% CI: −10.56, −6.10; p<0.001 and MD: −5.75; 95% CI: −7.75, −3.74; p<0.001) whereas, no significant difference was observed for hepatocellular cancer. Conclusions The present meta-analysis adds new evidence to the growing literature on the role of LINE-1 hypomethylation in human cancer and demonstrates that LINE-1 methylation levels were significantly lower in cancer patients than in control samples, especially in certain cancer types. This result was confirmed in tissue samples, both fresh/frozen or FFPE specimens, but not in blood. Further studies are needed to better clarify the role of LINE-1 methylation in specific subgroups, considering both cancer

  13. Proteomics analysis of tissue samples from patients with squamous cell carcinoma of the penis and positive to human papillomavirus

    Directory of Open Access Journals (Sweden)

    Leandro Koifman

    2015-08-01

    Full Text Available ABSTRACTPurpose:The aim of this study was to identify possible protein biomarkers and/or candidates for therapeutic targets in tissues of patients with SCCP, infected by HPV, applying one dimensional electrophoresis (1DE, followed by direct mass spectrometry (MS analysis.Materials and Methods:Tissues from 10 HPV positive patients with SCCP and from 10 patients with HPV negative non-tumorous penile foreskins were analyzed applying 1D electrophoresis, followed by analysis with direct mass spectrometry (MS.Results:Sixty-three different proteins were identified in the first group and 50 in the second group. Recognition was possible for 28 proteins exclusively detected in Group 1 and 21 proteins presented only in Group 2.Conclusion:Some proteins in the first group are directly involved in the development of other types of cancer, and therefore, suitable for analysis. Complement C3 protein is a strong candidate for evaluating SCCP patients.

  14. Morphometry in pathology: another look at diagnostic histopathology.

    Science.gov (United States)

    Collan, Y

    1984-11-01

    Histopathology is crucial for diagnostic and therapeutic decisions in many disease states. The created classifications, however, have not always allowed explicit recognition of diagnostic categories. So, considerable intra- and interobserver variations are possible under diagnostic circumstances. Better reproducibility can be reached by applying other than traditional methods. These include morphometry as well as special stains, electron microscopy and immunohistochemistry. Morphometry is basic in that in detects structural and morphological aberrations in samples prepared by other methods and without doubt brings the element of accuracy to support the diagnostic decision. Morphometric approach includes item classification and grading, point counting and intersection counting methods, and the use of various semiautomatic or automatic instruments. In statistical morphometry morphometrical parameters are collected from several disease cases. Such data reinforced with prognostic follow-ups are the basis for disease classification. Diagnostic morphometry, on the other hand, tries to study the sample of one individual and give relevant data for diagnostic decisions. Morphometric methods allow estimation of the limits of possible performance in histopathology, estimation of human performance in the diagnostic process and estimation of the influence of interfering human factors. Diagnostic decisions as well as decisions linked with therapy are basically probabilistic. Also the data collected by diagnostic morphometry should be linked with probabilistic estimates of the prognostic relevance of the findings. Education of histopathology should be developed to meet these demands, both at elementary medical level and at postgraduate medical level. Education of morphometry can be incorporated into the medical curricula as shown by the experience at the University of Kuopio where aspects of morphometry are taught during histopathology course and special postgraduate courses have

  15. Evaluation of tissue sampling methods used for MRI-detected contralateral breast lesions in the American College of Radiology Imaging Network 6667 trial.

    Science.gov (United States)

    DeMartini, Wendy B; Hanna, Lucy; Gatsonis, Constantine; Mahoney, Mary C; Lehman, Constance D

    2012-09-01

    The purpose of our study was to evaluate tissue sampling methods used for MRI-detected suspicious contralateral breast lesions in the American College of Radiology Imaging Network (ACRIN) 6667 trial. Breast MRI was performed at 25 institutions in 969 women who had a recent diagnosis of unilateral breast cancer and negative contralateral mammography and clinical breast examinations. Biopsy was recommended for MRI findings in 135 women, and 121 underwent sampling. Frequencies and positive biopsy rates of sampling methods used for initial diagnosis and imaging guidance techniques were calculated and compared. Sampling yielded 30 malignant and 91 benign results. Initial sampling used needle biopsy in 88 of 121 (72.7%) and surgical biopsy in 30 of 121 (24.8%) women. Surgical biopsy was excisional biopsy in 28 of 30 (93.3%) and mastectomy in two of 30 (6.7%). The remaining three of 121 (2.5%) women underwent mastectomy, but it was not documented whether this represented initial tissue sampling. Of imaging-guided procedures, 56 of 106 (52.8%) used MRI; 49 of 106 (46.2%), ultrasound; and one of 106 (1.0%), stereotaxis. MRI-guided sampling was with needle biopsy rather than wire-localized surgical biopsy in 33 of 56 (58.9%) women, whereas ultrasound used needle biopsy in 47 of 49 (95.9%). Positive biopsy rates of sampling methods were 20.5% for needle biopsy, 46.2% for excisional biopsy, and 0% for mastectomy. The majority of initial biopsies for MRI-detected contralateral breast lesions used needle biopsy rather than surgical biopsy. Contralateral surgery could have been avoided in most cases had needle biopsy been performed because most excisional biopsy and all mastectomy results were benign. MRI-guided biopsy was significantly more likely than ultrasound-guided sampling to use wire-localized surgical biopsy rather than needle biopsy.

  16. Wanted: dead or alive? Isotopic analysis (δ13C and δ15N) of Pygoscelis penguin chick tissues supports opportunistic sampling.

    Science.gov (United States)

    Vasil, Christopher A; Polito, Michael J; Patterson, William P; Emslie, Steven D

    2012-03-15

    Physiological stress and starvation have been shown to affect δ(13)C and δ(15)N isotope values and, given that animals often die from starvation, the cause of death may be an important factor to consider in stable isotope analyses of opportunistically collected samples. We addressed this issue by comparing tissue stable isotope values of living and deceased Adélie (Pygoscelis adeliae) and Chinstrap Penguin (P. antarctica) chicks collected from the same respective populations. No significant difference was found between living and deceased penguin chick feather, down, and toenail isotope values and both groups displayed similar isotopic trends between tissue types. In addition, similar relationships were observed between both species and across several seasons. Furthermore, sub-dermal adiposity and cause of death (starvation and/or predation) had no significant effect on the δ(13)C and δ(15)N values. Our findings suggest that tissues from deceased penguins can be isotopically representative of tissues obtained from the living population, despite the cause of death, and support the use of opportunistic sampling in stable isotope analyses. Copyright © 2012 John Wiley & Sons, Ltd.

  17. Principal component analysis of Draize eye Irritation tissue scores from 72 samples of 55 chemicals in the ECETOC data bank.

    Science.gov (United States)

    Lovell, D P

    1996-10-01

    The multivariate statistical method Principal Component Analysis (PCA) has been applied to a set of data from the ECETOC reference chemical data bank. PCA is a multivariate method that can be used to explore a complex data set. The results of the analysis show that most of the variability in the values for tissue damage scores for the 55 chemicals can be described by a single principal component which explains nearly 80% of the variability. This component is derived by giving approximately equal weight to each of the 18 individual measures made on the tissues over the 24-, 48- and 72-hr observation period. The principal component scores on the first component (PC I) are very highly correlated with the maximum individual weighted Draize scores or total Draize scores (TDS) derived using the Draize scoring method. A second principal component, describing about 7% of the variability, contrasts damage measured on the iris and cornea with that measured on the conjunctiva. Plots of principal component scores show the overall pattern of responses. In general, low measures of the TDS and a positive (PC I) score are associated with iris and conjunctival damage (damage to the iris was never recorded in the absence of damage to the conjunctiva). High TDS and negative PC I scores are associated with corneal and/or iris and conjunctiva damage. Plots of the principal component scores identify some chemicals that appear to cause unusual patterns of damage and identify some individual animals as having outlying or idiosyncratic responses. However, the analysis suggests that (i) there is only limited evidence for differential responses of different tissues and (ii) that attempts to identify alternative tests which predict specific types of tissue damage based on the results collected in a Draize test are likely to be unsuccessful. It indicates that further refinement of the results of the in vivo Draize test will not arise from more detailed analysis of the tissue scores but by

  18. Maintaining Breast Cancer Specimen Integrity and Individual or Simultaneous Extraction of Quality DNA, RNA, and Proteins from Allprotect-Stabilized and Nonstabilized Tissue Samples

    LENUS (Irish Health Repository)

    Mee, Blanaid C.

    2011-12-29

    The Saint James\\'s Hospital Biobank was established in 2008, to develop a high-quality breast tissue BioResource, as a part of the breast cancer clinical care pathway. The aims of this work were: (1) to ascertain the quality of RNA, DNA, and protein in biobanked carcinomas and normal breast tissues, (2) to assess the efficacy of AllPrep® (Qiagen) in isolating RNA, DNA, and protein simultaneously, (3) to compare AllPrep with RNEasy® and QIAamp® (both Qiagen), and (4) to examine the effectiveness of Allprotect® (Qiagen), a new tissue stabilization medium in preserving DNA, RNA, and proteins. One hundred eleven frozen samples of carcinoma and normal breast tissue were analyzed. Tumor and normal tissue morphology were confirmed by frozen sections. Tissue type, tissue treatment (Allprotect vs. no Allprotect), extraction kit, and nucleic acid quantification were analyzed by utilizing a 4 factorial design (SPSS PASW 18 Statistics Software®). QIAamp (DNA isolation), AllPrep (DNA, RNA, and Protein isolation), and RNeasy (RNA isolation) kits were assessed and compared. Mean DNA yield and A260\\/280 values using QIAamp were 33.2 ng\\/μL and 1.86, respectively, and using AllPrep were 23.2 ng\\/μL and 1.94. Mean RNA yield and RNA Integrity Number (RIN) values with RNeasy were 73.4 ng\\/μL and 8.16, respectively, and with AllPrep were 74.8 ng\\/μL and 7.92. Allprotect-treated tissues produced higher RIN values of borderline significance (P=0.055). No discernible loss of RNA stability was detected after 6 h incubation of stabilized or nonstabilized tissues at room temperature or 4°C or in 9 freeze-thaw cycles. Allprotect requires further detailed evaluation, but we consider AllPrep to be an excellent option for the simultaneous extraction of RNA, DNA, and protein from tumor and normal breast tissues. The essential presampling procedures that maintain the diagnostic integrity of pathology specimens do not appear to compromise the quality of molecular isolates.

  19. Detection of Lawsonia intracellularis in formalinfixed Porcine Intestinal Tissue Samples: Comparison of Immunofluorescense and In-situ Hybridization, and Evaluation of the Effectf of Controlled Autolysis

    DEFF Research Database (Denmark)

    Jensen, Tim Kåre; Boesen, H. T.; Vigre, Håkan;

    2010-01-01

    Two methods, an immunofluorescence assay (IFA; with a Lawsonia intracellularis-specific monoclonal antibody) and fluorescent in-situ hybridization (FISH; with a specific oligonucleotide probe targeting 16S ribosomal RNA of the bacterium), were compared for their ability to detect L. intracellularis...... (the cause of porcine proliferative enteritis [PE]) in formalin-fixed samples of intestinal tissue. Of 69 intestinal samples with gross lesions of PE, 63 were positive by both FISH and IFA, but six were positive only by IFA. This indicated that the sensitivity of FISH was 91% that of IFA. However, both...

  20. [Accuracy of PCR for the detection of bacterial and fungal DNA in the blood and tissue samples of experimentally infected rabbits].

    Science.gov (United States)

    Fouad, Ali Adil; Kalkancı, Ayşe

    2012-10-01

    Direct demonstration of bacterial and/or fungal nucleic acids in the clinical samples of patients with blood stream infections is crucial in terms of rapid diagnosis, early and accurate therapy and patient management. This study was aimed to determine the presence of bacteria and fungi by polymerase chain reaction (PCR) in the clinical samples of experimental sepsis induced animals, to compare the results with culture and to evaluate the efficiency of PCR in the discrimination of bacteremia and fungemia. A total of 12 rabbits experimentally infected with standard strains of Staphylococcus aureus, Escherichia coli, Aspergillus fumigatus and Candida albicans to generate bacteremia (n= 4), fungemia (n= 4) and polymicrobial blood stream infection (n= 4), were included in the study. A total of 63 specimens of which 27 were blood and 36 were tissue (12 spleen, 12 liver, 12 kidney) samples were collected at 24, 48, 72 and 96th hours of infection. Uninfected healthy rabbits (n= 4), colony suspensions of standard bacterial and fungal strains (n= 15) and human blood samples contaminated with standard bacterial and fungal strains (n= 10) were used as controls. Microbial DNAs were searched by using real-time PCR in all the samples, and quantitative cultures were performed simultaneously. Gram-positive and gram-negative PCR protocols were performed for the samples of bacteremic animals, whereas panfungal PCR, Aspergillus and Candida PCR protocols were performed for the samples of animals with fungemia. All of those PCR protocols were applied separately for the samples of polymicrobial blood stream infection cases. Culture positivity was detected in 8 (29.6%) of the blood samples and bacterial and/or fungal DNAs were demonstrated in 20 (74%) of the blood samples by PCR. Microbial DNAs were also detected in 32 (89%) of 36 tissue samples (11 spleen, 11 liver, 10 kidney). Sensitivity rates of culture method to detect bacteremia and fungemia were 30% and 21.7%, respectively, whereas

  1. Numerical and structural genomic aberrations are reliably detectable in tissue microarrays of formalin-fixed paraffin-embedded tumor samples by fluorescence in-situ hybridization.

    Directory of Open Access Journals (Sweden)

    Heike Horn

    Full Text Available Few data are available regarding the reliability of fluorescence in-situ hybridization (FISH, especially for chromosomal deletions, in high-throughput settings using tissue microarrays (TMAs. We performed a comprehensive FISH study for the detection of chromosomal translocations and deletions in formalin-fixed and paraffin-embedded (FFPE tumor specimens arranged in TMA format. We analyzed 46 B-cell lymphoma (B-NHL specimens with known karyotypes for translocations of IGH-, BCL2-, BCL6- and MYC-genes. Locus-specific DNA probes were used for the detection of deletions in chromosome bands 6q21 and 9p21 in 62 follicular lymphomas (FL and six malignant mesothelioma (MM samples, respectively. To test for aberrant signals generated by truncation of nuclei following sectioning of FFPE tissue samples, cell line dilutions with 9p21-deletions were embedded into paraffin blocks. The overall TMA hybridization efficiency was 94%. FISH results regarding translocations matched karyotyping data in 93%. As for chromosomal deletions, sectioning artefacts occurred in 17% to 25% of cells, suggesting that the proportion of cells showing deletions should exceed 25% to be reliably detectable. In conclusion, FISH represents a robust tool for the detection of structural as well as numerical aberrations in FFPE tissue samples in a TMA-based high-throughput setting, when rigorous cut-off values and appropriate controls are maintained, and, of note, was superior to quantitative PCR approaches.

  2. Numerical and structural genomic aberrations are reliably detectable in tissue microarrays of formalin-fixed paraffin-embedded tumor samples by fluorescence in-situ hybridization.

    Science.gov (United States)

    Horn, Heike; Bausinger, Julia; Staiger, Annette M; Sohn, Maximilian; Schmelter, Christopher; Gruber, Kim; Kalla, Claudia; Ott, M Michaela; Rosenwald, Andreas; Ott, German

    2014-01-01

    Few data are available regarding the reliability of fluorescence in-situ hybridization (FISH), especially for chromosomal deletions, in high-throughput settings using tissue microarrays (TMAs). We performed a comprehensive FISH study for the detection of chromosomal translocations and deletions in formalin-fixed and paraffin-embedded (FFPE) tumor specimens arranged in TMA format. We analyzed 46 B-cell lymphoma (B-NHL) specimens with known karyotypes for translocations of IGH-, BCL2-, BCL6- and MYC-genes. Locus-specific DNA probes were used for the detection of deletions in chromosome bands 6q21 and 9p21 in 62 follicular lymphomas (FL) and six malignant mesothelioma (MM) samples, respectively. To test for aberrant signals generated by truncation of nuclei following sectioning of FFPE tissue samples, cell line dilutions with 9p21-deletions were embedded into paraffin blocks. The overall TMA hybridization efficiency was 94%. FISH results regarding translocations matched karyotyping data in 93%. As for chromosomal deletions, sectioning artefacts occurred in 17% to 25% of cells, suggesting that the proportion of cells showing deletions should exceed 25% to be reliably detectable. In conclusion, FISH represents a robust tool for the detection of structural as well as numerical aberrations in FFPE tissue samples in a TMA-based high-throughput setting, when rigorous cut-off values and appropriate controls are maintained, and, of note, was superior to quantitative PCR approaches.

  3. Body fluid and tissue analysis using filter paper sampling support prior to LC-MS/MS: application to fatal overdose with colchicine.

    Science.gov (United States)

    Lauer, Estelle; Widmer, Christèle; Versace, François; Staub, Christian; Mangin, Patrice; Sabatasso, Sara; Augsburger, Marc; Déglon, Julien

    2013-01-01

    Because of the various matrices available for forensic investigations, the development of versatile analytical approaches allowing the simultaneous determination of drugs is challenging. The aim of this work was to assess a liquid chromatography-tandem mass spectrometry (LC-MS/MS) platform allowing the rapid quantification of colchicine in body fluids and tissues collected in the context of a fatal overdose. For this purpose, filter paper was used as a sampling support and was associated with an automated 96-well plate extraction performed by the LC autosampler itself. The developed method features a 7-min total run time including automated filter paper extraction (2 min) and chromatographic separation (5 min). The sample preparation was reduced to a minimum regardless of the matrix analyzed. This platform was fully validated for dried blood spots (DBS) in the toxic concentration range of colchicine. The DBS calibration curve was applied successfully to quantification in all other matrices (body fluids and tissues) except for bile, where an excessive matrix effect was found. The distribution of colchicine for a fatal overdose case was reported as follows: peripheral blood, 29 ng/ml; urine, 94 ng/ml; vitreous humour and cerebrospinal fluid, paper is usually employed for DBS, we report here the extension of this alternative sampling support to the analysis of other body fluids and tissues. The developed platform represents a rapid and versatile approach for drug determination in multiple forensic media.

  4. Pre-Analytical Considerations for Successful Next-Generation Sequencing (NGS: Challenges and Opportunities for Formalin-Fixed and Paraffin-Embedded Tumor Tissue (FFPE Samples

    Directory of Open Access Journals (Sweden)

    Gladys Arreaza

    2016-09-01

    Full Text Available In cancer drug discovery, it is important to investigate the genetic determinants of response or resistance to cancer therapy as well as factors that contribute to adverse events in the course of clinical trials. Despite the emergence of new technologies and the ability to measure more diverse analytes (e.g., circulating tumor cell (CTC, circulating tumor DNA (ctDNA, etc., tumor tissue is still the most common and reliable source for biomarker investigation. Because of its worldwide use and ability to preserve samples for many decades at ambient temperature, formalin-fixed, paraffin-embedded tumor tissue (FFPE is likely to be the preferred choice for tissue preservation in clinical practice for the foreseeable future. Multiple analyses are routinely performed on the same FFPE samples (such as Immunohistochemistry (IHC, in situ hybridization, RNAseq, DNAseq, TILseq, Methyl-Seq, etc.. Thus, specimen prioritization and optimization of the isolation of analytes is critical to ensure successful completion of each assay. FFPE is notorious for producing suboptimal DNA quality and low DNA yield. However, commercial vendors tend to request higher DNA sample mass than what is actually required for downstream assays, which restricts the breadth of biomarker work that can be performed. We evaluated multiple genomics service laboratories to assess the current state of NGS pre-analytical processing of FFPE. Significant differences in pre-analytical capabilities were observed. Key aspects are highlighted and recommendations are made to improve the current practice in translational research.

  5. MALDI mass spectrometry imaging: A cutting-edge tool for fundamental and clinical histopathology.

    Science.gov (United States)

    Longuespée, Rémi; Casadonte, Rita; Kriegsmann, Mark; Pottier, Charles; Picard de Muller, Gaël; Delvenne, Philippe; Kriegsmann, Jörg; De Pauw, Edwin

    2016-07-01

    Histopathological diagnoses have been done in the last century based on hematoxylin and eosin staining. These methods were complemented by histochemistry, electron microscopy, immunohistochemistry (IHC), and molecular techniques. Mass spectrometry (MS) methods allow the thorough examination of various biocompounds in extracts and tissue sections. Today, mass spectrometry imaging (MSI), and especially matrix-assisted laser desorption ionization (MALDI) imaging links classical histology and molecular analyses. Direct mapping is a major advantage of the combination of molecular profiling and imaging. MSI can be considered as a cutting edge approach for molecular detection of proteins, peptides, carbohydrates, lipids, and small molecules in tissues. This review covers the detection of various biomolecules in histopathological sections by MSI. Proteomic methods will be introduced into clinical histopathology within the next few years.

  6. Real-time PCR strategy for parasite quantification in blood and tissue samples of experimental Trypanosoma cruzi infection.

    Science.gov (United States)

    Caldas, Sérgio; Caldas, Ivo Santana; Diniz, Lívia de Figueiredo; Lima, Wanderson Geraldo de; Oliveira, Riva de Paula; Cecílio, Alzira Batista; Ribeiro, Isabela; Talvani, André; Bahia, Maria Terezinha

    2012-09-01

    The lack of an accurate diagnosis has been a serious obstacle to the advancement of the anti-Trypanosoma cruzi chemotherapy and long-term infection can result in different health risks to human. PCRs are alternative methods, more sensitive than conventional parasitological techniques, which due to their low sensitivities are considered unsuitable for these purposes. The aim of this study was to investigate a sensitive diagnostic strategy to quantify blood and cardiac tissues parasites based on real-time PCR tools during acute and chronic phases of murine Chagas disease, as well as to monitor the evolution of infection in those mice under specific treatment. In parallel, fresh blood examination, immunological analysis and quantification of cardiac inflammation were also performed to confront and improve real-time PCR data. Similar profiles of parasitemia curves were observed in both quantification techniques during the acute phase of the infection. In contrast, parasites could be quantified only by real-time PCR at 60 and 120 days of infection. In cardiac tissue, real-time PCR detected T. cruzi DNA in 100% of infected mice, and using this tool a significant Pearson correlation between parasite load in peripheral blood and in cardiac tissue during acute and chronic phases was observed. Levels of serum CCL2, CCL5 and nitric oxide were coincident with parasite load but focal and diffuse mononuclear infiltrates was observed, even with significant (pblood and cardiac muscle at the treatment period, but after the end of chemotherapy an increase of parasitism was detected. Interestingly, inflammatory mediators levels and heart inflammation intensity had similar evolution to the parasite load, in the group of animals treated. Taken together, our data show that real-time PCR strategy used was suitable for studies of murine T. cruzi infection and may prove useful in investigations involving experimental chemotherapy of the disease and the benefits of treatment in relation to

  7. Multi-scale occupancy approach to estimate Toxoplasma gondii prevalence and detection probability in tissues: an application and guide for field sampling.

    Science.gov (United States)

    Elmore, Stacey A; Huyvaert, Kathryn P; Bailey, Larissa L; Iqbal, Asma; Su, Chunlei; Dixon, Brent R; Alisauskas, Ray T; Gajadhar, Alvin A; Jenkins, Emily J

    2016-08-01

    Increasingly, birds are recognised as important hosts for the ubiquitous parasite Toxoplasma gondii, although little experimental evidence exists to determine which tissues should be tested to maximise the detection probability of T. gondii. Also, Arctic-nesting geese are suspected to be important sources of T. gondii in terrestrial Arctic ecosystems, but the parasite has not previously been reported in the tissues of these geese. Using a domestic goose model, we applied a multi-scale occupancy framework to demonstrate that the probability of detection of T. gondii was highest in the brain (0.689, 95% confidence interval=0.486, 0.839) and the heart (0.809, 95% confidence interval=0.693, 0.888). Inoculated geese had an estimated T. gondii infection probability of 0.849, (95% confidence interval=0.643, 0.946), highlighting uncertainty in the system, even under experimental conditions. Guided by these results, we tested the brains and hearts of wild Ross's Geese (Chen rossii, n=50) and Lesser Snow Geese (Chen caerulescens, n=50) from Karrak Lake, Nunavut, Canada. We detected 51 suspected positive tissue samples from 33 wild geese using real-time PCR with melt-curve analysis. The wild goose prevalence estimates generated by our multi-scale occupancy analysis were higher than the naïve estimates of prevalence, indicating that multiple PCR repetitions on the same organs and testing more than one organ could improve T. gondii detection. Genetic characterisation revealed Type III T. gondii alleles in six wild geese and Sarcocystis spp. in 25 samples. Our study demonstrates that Arctic nesting geese are capable of harbouring T. gondii in their tissues and could transport the parasite from their southern overwintering grounds into the Arctic region. We demonstrate how a multi-scale occupancy framework can be used in a domestic animal model to guide resource-limited sample collection and tissue analysis in wildlife. Secondly, we confirm the value of traditional occupancy in

  8. The application of pancuronium bromide (Pavulon) forensic analyses to tissue samples from an "Angel of Death" investigation.

    Science.gov (United States)

    Andresen, Brian D; Alcaraz, Armando; Grant, Patrick M

    2005-01-01

    The case report of a serial killer who worked at several hospitals as a respiratory therapist is presented. The suspect was initially labeled a benevolent Angel of Death who ended the suffering of elderly patients through mercy killing. However, his subsequently declared motive for homicide was very different from other similar cases in medical settings. The application of new analysis techniques for the detection of pancuronium bromide in a series of aged exhumation tissues gave positive results and led to the resultant conviction of the therapist.

  9. Proteoglycan and proteome profiling of central human pulmonary fibrotic tissue utilizing miniaturized sample preparation: a feasibility study

    DEFF Research Database (Denmark)

    Malmström, Johan; Larsen, Kristoffer; Hansson, Lennart;

    2002-01-01

    The objective of this study was to isolate fibrotic cells from human lung biopsies taken from different central pulmonary locations. A comparison was made of cell morphology, proteoglycan- and protein-expression in mesenchymal cell cultures obtained from human bronchial biopsies from patients...... tissue; one of contractile type with lamellipodia that facilitate migration and a second cell type with an increased cell size, which most likely is of a synthetic phenotype. This is the first evidence of alterations in the proteoglycan expression pattern of versican, perlecan, biglycan and decorin which...

  10. Histopathologic ear findings of syphilis: a temporal bone study.

    Science.gov (United States)

    Hızlı, Ömer; Hızlı, Pelin; Kaya, Serdar; Monsanto, Rafael da Costa; Paparella, Michael M; Cureoglu, Sebahattin

    2016-09-01

    To the best of our knowledge, histopathologic studies of syphilitic ears have generally focused on hydropic changes; so far, no such studies have investigated peripheral vestibular otopathology using differential interference contrast microscopy, in patients with syphilis. For this study, we examined 13 human temporal bone samples from 8 patients with a history of syphilis. Using conventional light microscopy, we performed qualitative histopathologic assessment. In addition, using differential interference contrast microscopy, we performed type I and type II vestibular hair cell counts on each vestibular sense organ with minimal autolysis; in which the neuroepithelium was oriented perpendicular to the plane of section. We then compared vestibular hair cell densities (cells per 0.01 mm² surface area) in the syphilis group vs. the control group. In the syphilis group, we observed precipitate in the endolymphatic or perilymphatic spaces in 1 (7.7 %) of the samples and endolymphatic hydrops in eight (61.5 %) of the samples. Hydrops involved the cochlea (four samples) and/or saccule (four samples). In addition, the syphilis group experienced a significant loss of type II vestibular hair cells in the maculae of the utricle and saccule, and in the cristae of the lateral and posterior semicircular canals, as compared with the control group (P < 0.05).

  11. Histopathology of explanted AlphaCor due to keratoprosthesis extrusion.

    Science.gov (United States)

    Caporossi, Aldo; Mazzotta, Cosimo; Balestrazzi, Angelo; Occhini, Rossella; Toti, Paolo; Traversi, Claudio

    2006-07-01

    AlphaCor keratoprosthesis (KPro) is a new-concept poly (2-hydroxyethyl methacrylate) one-piece KPro that makes possible a two-step implantation technique easy to perform with a short learning curve. In literature an 18% incidence of AlphaCor removal due to melting complications is reported. The histopathology of corneal tissue removed during a re-operation while bearing an AlphaCor KPro has previously been described in the literature only in one report. Herein, the first histological features of an AlphaCor-corneal complex explanted because of KPro extrusion is described. The histopathology of the AlphaCor-corneal complex is characterized by mild inflammation in the corneal tissues, limited to the region surrounding the anteriorized and extruded part of the KPro. It is not possible to fully understand the mechanisms that trigger the device extrusion. One possible explanation could be a dislocation of the prosthesis in the corneal pocket due to the untied fixation stitch. Another explanation could be a foreign body reaction induced by KPro.

  12. Active immunization against GnRH in ovine (Ovis aries: testicular morphometry, histopathology and endocrine responses in prepubertal ram lambs

    Directory of Open Access Journals (Sweden)

    Gutierrez-Reinoso MA

    2017-08-01

    Full Text Available The main objective of the present research was to assess the effects of an active immunization against GnRH on male reproductive function as an alternative to surgical sterilization evaluating testicular morphometry, histopathological alterations and plasma gonadotropin levels in prepubertal ram lambs. Dorper ram lambs (age= 2 months; control group, n= 5; treatment group, n=15 were immunised by using an anti-GnRH vaccine (two administrations spaced 15d developed by linking a GnRH homologous molecule to tetanus toxoid (clostridial toxoid with aluminum hydroxide as coadjuvant. To determine the effectiveness of the vaccination protocol, testicular morphometry was evaluated (length, width, mean volume and total volume together with histopathological alterations in testicular tissue samples (seminiferous tubule diameters, spermatogonia per testis and sperm presence and endocrine responses (ELISA from blood samples (FSH, LH, testosterone and cortisol plasma levels. Morphometric parameters (testicular length, width and volume were significantly reduced in vaccinated animals with respect to the control group (p0.05. In conclusion, prepubertal active immunization against GnRH led to marked differences on testes morphometry and activity in ovine species, however, it did not affect endocrine levels nor spermatogenesis in all individuals studied showing a partial vaccination effect in some of them. Thus, taking into account the heterogeneous dysfunctional responses obtained, different vaccination assays and strategies should be tested before active immunization against GnRH is considered as a viable alternative to conventional surgical sterilization.

  13. Histopathological pattern of abnormal uterine bleeding in endometrial biopsies.

    Science.gov (United States)

    Vaidya, S; Lakhey, M; Vaidya, S; Sharma, P K; Hirachand, S; Lama, S; KC, S

    2013-03-01

    Abnormal uterine bleeding is a common presenting complaint in gyanecology out patient department. Histopathological evaluation of the endometrial samples plays a significant role in the diagnosis of abnormal uterine bleeding. This study was carried out to determine the histopathological pattern of the endometrium in women of various age groups presenting with abnormal uterine bleeding. Endometrial biopsies and curettings of patients presenting with abnormal uterine bleeding was retrospectively studied. A total of 403 endometrial biopsies and curettings were analyzed. The age of the patients ranged from 18 to 70 years. Normal cyclical endometrium was seen in 165 (40.94%) cases, followed by 54 (13.40%) cases of disordered proliferative endometrium and 44 (10.92%) cases of hyperplasia. Malignancy was seen in 10 (2.48%) cases. Hyperplasia and malignancy were more common in the perimenopausal and postmenopausal age groups. Histopathological examination of endometrial biopsies and curettings in patients presenting with abnormal uterine bleeding showed a wide spectrum of changes ranging from normal endometrium to malignancy. Endometrial evaluation is specially recommended in women of perimenopausal and postmenopausal age groups presenting with AUB, to rule out a possibility of any preneoplastic condition or malignancy.

  14. Topography of Genetic Loci in Tissue Samples: Towards New Diagnostic Tool Using Interphase FISH and High-Resolution Image Analysis Techniques

    Directory of Open Access Journals (Sweden)

    I. Koutná

    2000-01-01

    Full Text Available Using single and dual colour fluorescence in situ hybridisation (FISH combined with image analysis techniques the topographic characteristics of genes and centromeres in nuclei of human colon tissue cells were investigated. The distributions of distances from the centre‐of‐nucleus to genes (centromeres and from genes to genes (centromeres to centromeres were studied in normal colon tissue cells found in the neighbourhood of tumour samples, in tumour cell line HT‐29 and in promyelocytic HL‐60 cell line for comparison. Our results show that the topography of genetic loci determined in 3D‐fixed cell tissue corresponds to that obtained for 2D‐fixed cells separated from the tissue. The distributions of the centre‐of‐nucleus to gene (centromere distances and gene to gene (centromere to centromere distances and their average values are different for various genetic loci but similar for normal colon tissue cells, HT‐29 colon tumour cell line and HL‐60 promyelocytic cell line. It suggests that the arrangement of genetic loci in cell nucleus is conserved in different types of human cells. The investigations of trisomic loci in HT‐29 cells revealed that the location of the third genetic element is not different from the location of two homologues in diploid cells. We have shown that the topographic parameters used in our experiments for different genetic elements are not tissue or tumour specific. In order to validate high‐resolution cytometry for oncology, further investigations should include more precise parameters reflecting the state of chromatin in the neighbourhood of critical oncogenes or tumour suppresser genes.

  15. Synchrotron refractive-index microradiography of human liver cancer tissue

    Institute of Scientific and Technical Information of China (English)

    TONG Yongpeng; ZHANG Guilin; LI Yan; HWU Yeukuang; TSAI Wenli; JE Jung Ho; Margaritondo G.; YUAN Dong

    2005-01-01

    Three human liver tissue samples (~5 mm × 40 mm × 20 mm) were excised from a cancer patient's liver during surgery. The microradiology analysis was performed with a non-standard approach on a synchrotron. High-resolution refractive-index edge-enhanced microradiographs that cover a larger volume of the liver tissue sample were obtained. The cancer tissue and normal tissue could be clearly identified and distinguished based on their different textures. Furthermore, new blood vessel hyperplasia was found near the cancer area. Blood vessels with a diameter smaller than 20 μm could be identified. These findings were fully consistent with the histopathological examination of the same area. Microradiographs of the newly formed blood vessels at different angles were also obtained. This result shows that it is possible to further develop this approach into a technique of microradiographic imaging for clinic diagnosis of liver cancer at the early stage.

  16. Histopathology reveals correlative and unique phenotypes in a high-throughput mouse phenotyping screen

    Directory of Open Access Journals (Sweden)

    Hibret A. Adissu

    2014-05-01

    Full Text Available The Mouse Genetics Project (MGP at the Wellcome Trust Sanger Institute aims to generate and phenotype over 800 genetically modified mouse lines over the next 5 years to gain a better understanding of mammalian gene function and provide an invaluable resource to the scientific community for follow-up studies. Phenotyping includes the generation of a standardized biobank of paraffin-embedded tissues for each mouse line, but histopathology is not routinely performed. In collaboration with the Pathology Core of the Centre for Modeling Human Disease (CMHD we report the utility of histopathology in a high-throughput primary phenotyping screen. Histopathology was assessed in an unbiased selection of 50 mouse lines with (n=30 or without (n=20 clinical phenotypes detected by the standard MGP primary phenotyping screen. Our findings revealed that histopathology added correlating morphological data in 19 of 30 lines (63.3% in which the primary screen detected a phenotype. In addition, seven of the 50 lines (14% presented significant histopathology findings that were not associated with or predicted by the standard primary screen. Three of these seven lines had no clinical phenotype detected by the standard primary screen. Incidental and strain-associated background lesions were present in all mutant lines with good concordance to wild-type controls. These findings demonstrate the complementary and unique contribution of histopathology to high-throughput primary phenotyping of mutant mice.

  17. Histopathology reveals correlative and unique phenotypes in a high-throughput mouse phenotyping screen.

    Science.gov (United States)

    Adissu, Hibret A; Estabel, Jeanne; Sunter, David; Tuck, Elizabeth; Hooks, Yvette; Carragher, Damian M; Clarke, Kay; Karp, Natasha A; Newbigging, Susan; Jones, Nora; Morikawa, Lily; White, Jacqueline K; McKerlie, Colin

    2014-05-01

    The Mouse Genetics Project (MGP) at the Wellcome Trust Sanger Institute aims to generate and phenotype over 800 genetically modified mouse lines over the next 5 years to gain a better understanding of mammalian gene function and provide an invaluable resource to the scientific community for follow-up studies. Phenotyping includes the generation of a standardized biobank of paraffin-embedded tissues for each mouse line, but histopathology is not routinely performed. In collaboration with the Pathology Core of the Centre for Modeling Human Disease (CMHD) we report the utility of histopathology in a high-throughput primary phenotyping screen. Histopathology was assessed in an unbiased selection of 50 mouse lines with (n=30) or without (n=20) clinical phenotypes detected by the standard MGP primary phenotyping screen. Our findings revealed that histopathology added correlating morphological data in 19 of 30 lines (63.3%) in which the primary screen detected a phenotype. In addition, seven of the 50 lines (14%) presented significant histopathology findings that were not associated with or predicted by the standard primary screen. Three of these seven lines had no clinical phenotype detected by the standard primary screen. Incidental and strain-associated background lesions were present in all mutant lines with good concordance to wild-type controls. These findings demonstrate the complementary and unique contribution of histopathology to high-throughput primary phenotyping of mutant mice.

  18. Automated Liquid Microjunction Surface Sampling-HPLC-MS/MS Analysis of Drugs and Metabolites in Whole-Body Thin Tissue Sections

    Energy Technology Data Exchange (ETDEWEB)

    Kertesz, Vilmos [ORNL; Van Berkel, Gary J [ORNL

    2013-01-01

    A fully automated liquid extraction-based surface sampling system utilizing a commercially available autosampler coupled to high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) detection is reported. Discrete spots selected for droplet-based sampling and automated sample queue generation for both the autosampler and MS were enabled by using in-house developed software. In addition, co-registration of spatially resolved sampling position and HPLC-MS information to generate heatmaps of compounds monitored for subsequent data analysis was also available in the software. The system was evaluated with whole-body thin tissue sections from propranolol dosed rat. The hands-free operation of the system was demonstrated by creating heatmaps of the parent drug and its hydroxypropranolol glucuronide metabolites with 1 mm resolution in the areas of interest. The sample throughput was approximately 5 min/sample defined by the time needed for chromatographic separation. The spatial distributions of both the drug and its metabolites were consistent with previous studies employing other liquid extraction-based surface sampling methodologies.

  19. NMR relaxation times of trabecular bone-reproducibility, relationships to tissue structure and effects of sample freezing

    Energy Technology Data Exchange (ETDEWEB)

    Prantner, Viktoria; Isaksson, Hanna; Nissi, Mikko J; Jurvelin, Jukka S [Department of Physics and Mathematics, University of Eastern Finland, PO Box 1627, 70211 Kuopio (Finland); Naervaeinen, Johanna; Groehn, Olli H J [Department of Neurobiology, A I Virtanen Institute for Molecular Sciences, University of Eastern Finland, PO Box 1627, 70211 Kuopio (Finland); Lammentausta, Eveliina [Department of Diagnostic Radiology, Oulu University Hospital, PO Box 50, 90029 OYS, Oulu (Finland); Avela, Janne, E-mail: hanna.isaksson@uef.f [Department of Biology of Physical Activity, University of Jyvaeskylae, PO Box 35, 40014 Jyvaeskylae (Finland)

    2010-12-07

    Nuclear magnetic resonance (NMR) spectroscopy provides a potential tool for non-invasive evaluation of the trabecular bone structure. The objective of this study was to determine the reproducibility of the NMR relaxation parameters (T{sub 2}, Carr-Purcel-T{sub 2}, T{sub 1}{rho}) for fat and water and relate those to the structural parameters obtained by micro-computed tomography ({mu}CT). Especially, we aimed to evaluate the effect of freezing on the relaxation parameters. For storing bone samples, freezing is the standard procedure during which the biochemical and cellular organization of the bone marrow may be affected. Bovine trabecular bone samples were stored at -20 {sup 0}C for 7 days and measured by NMR spectroscopy before and after freezing. The reproducibility of NMR relaxation parameters, as expressed by the coefficient of variation, ranged from 3.1% to 27.9%. In fresh samples, some correlations between NMR and structural parameters (Tb.N, Tb.Sp) were significant (e.g. the relaxation rate for T{sub 2} of fat versus Tb.Sp: r = -0.716, p < 0.01). Freezing did not significantly change the NMR relaxation times but the correlations between relaxation parameters and the {mu}CT structural parameters were not statistically significant after freezing, suggesting some nonsystematic alterations of the marrow structure. Therefore, the use of frozen bone samples for NMR relaxation studies may provide inferior information about the trabecular bone structure.

  20. Cell-type-specific expression of STAT transcription factors in tissue samples from patients with lymphocytic thyroiditis.

    Science.gov (United States)

    Staab, Julia; Barth, Peter J; Meyer, Thomas

    2012-09-01

    Expression of cytokine-regulated signal transducer and activator of transcription (STAT) proteins was histochemically assessed in patients diagnosed as having Hashimoto's disease or focal lymphocytic thyroiditis (n = 10). All surgical specimens showed histological features of lymphocytic thyroiditis, including a diffuse infiltration with mononuclear cells and an incomplete loss of thyroid follicles, resulting in the destruction of glandular tissue architecture. Immunohistochemical analysis demonstrated differential expression patterns of the various members of the STAT transcription factors examined, indicating that each member of this conserved protein family has its distinct functions in the development of the disease. Using an antibody that specifically recognized the phosphorylated tyrosine residue in position 701, we detected activated STAT1 dimers in numerous germinal macrophages and infiltrating lymphocytes as well as in oncocytes. In contrast, STAT3 expression was restricted to epithelial cells and showed a clear colocalization with the antiapoptotic protein Bcl-2. Moreover, expression of phospho-STAT3 was associated with low levels of stromal fibrosis, suggesting that STAT3 serves as a protective factor in the remodeling of the inflamed thyroid gland. Phospho-STAT5 immunoreactivity was detected in numerous infiltrating cells of hematopoietic origin and, additionally, in hyperplastic follicular epithelia. This tissue distribution demonstrated that activated STAT5 molecules participate in both lymphocytopoiesis and possibly also in the buildup of regenerating thyroid follicles. Taken together, the cell-type-specific expression patterns of STAT proteins in human lymphocytic thyroiditis reflect their distinct and partially antagonistic roles in orchestrating the balance between degenerating and regenerating processes within a changing cytokine environment.

  1. Diet, lifestyle, heritable factors and colorectal carcinogenesis: associations with histopathological and molecular endpoints

    NARCIS (Netherlands)

    Wark, P.A.

    2007-01-01

    Background: Diet, lifestyle and heritable factors have been related to colorectal cancer risk; to date, their relevance to the overall scope of colorectal carcinogenesis, has not been clearly established.Aim and Methods: To evaluate whether distinguishing colorectal tissue by its histopathological a

  2. Diet, lifestyle, heritable factors and colorectal carcinogenesis: associations with histopathological and molecular endpoints

    NARCIS (Netherlands)

    Wark, P.A.

    2007-01-01

    Background: Diet, lifestyle and heritable factors have been related to colorectal cancer risk; to date, their relevance to the overall scope of colorectal carcinogenesis, has not been clearly established.Aim and Methods: To evaluate whether distinguishing colorectal tissue by its histopathological

  3. Differential Gene Expression of BRCA1,ERBB2 and TP53 biomarkers between Human Breast Tissue and Peripheral Blood Samples of Breast Cancer.

    Science.gov (United States)

    Zghair, Abdulrazzaq Neamah; Sinha, Deepak Kumar; Kassim, Arkan; Alfaham, Mohmmad; Sharma, Anil K

    2016-01-01

    Breast cancer is a most common malignancy especially in Iraqi women accounting for high morbidity and mortality. Mutations in BRCA1 gene is one of the important genetic predisposing factors inbreast cancer. Similarly ERBB2 and TP53 are also key prognostic markers in breast cancer treatment.We were interested to explore the gene expression profiles of BRCA1, ERBB2 and TP53 in breast cancer women patients from Iraq so as to assess the potential of such markers in breast cancer treatment. The mRNA levels were significantly over-expressed in tumor tissues in comparison to normal ones with p values (pTP53 and benign tissue samples as well. However in blood samples, no considerable expression of these markers was observed. Out of three selected genes, ERBB2 expression was significantly expressed in comparison to BRCA1 and TP53 in cancer tissue. Mutation analysis of BRCA1, ERBB2 and TP53 has been made to find out the region most susceptible to mutations in these genes The BRCA1 exon 11, ERBB2 16 and TP53 exon 5 displayed increased chances of having mutations. We can conclude from the study that differential gene expression of BRCA1, ERBB2 and TP53 at mRNA levels may act as a diagnostic marker of circulating tumor cells having important prognostic value in breast cancer patients.

  4. Is routine histopathology of tonsil specimen necessary?

    Directory of Open Access Journals (Sweden)

    Agida S Adoga

    2011-01-01

    Full Text Available Background: Tonsillar diseases are common in paediatric and adult otolaryngological practice. These diseases require tonsillectomy. Specimens are subjected to histopathology routinely in my institution for fear of infections and tumour without consideration for risk factors. The financial burden is on the patients and waste of histopathologist′s man hour because other specimens are left un-attended. This study aims to find out the necessity of routine histopathology of tonsil specimens. Materials and Methods : A 2 year retrospective review of the histopathological results of two (paediatric and adult groups of 61 patients managed for tonsillar diseases at the ENT UNIT of Jos University Teaching Hospital from July 2005 to June, 2007. Data extracted included biodata, clinical features and histopathological diagnosis. Result : The 61 patients comprise 35 children and 26 adults. The youngest and oldest paediatric patients were 1 year and 3 months and 16 years respectively, a range of 1 year 3 months to 16 years. The youngest and oldest adults were 17 and 50 years with a range of 17-50 years. Groups mean ages were 5.1 and 28.5 years. The gender ratios were 1:2.7 and 1:1.9 respectively. One adult was HIV positive. The histopathological diagnosis were chronic nonspecific tonsillitis in 10(16.6%, follicular tonsillitis in 23(38.3%, chronic suppurative tonsillitis in 10(16.6%, lymphoid hyperplasia in 18(30.0% and lymphoma in 1(1.0% respectively. Conclusion : Histopathologic request for tonsillectomy specimens should be based on certain risk factors with consideration of the cost to patients and to spare the histopathologist′s man hour.

  5. Histopathological study of congenital aortic valve malformations in 32 children

    Institute of Scientific and Technical Information of China (English)

    HUANG Ping; WANG Hongwei; LI Yanping; CHENG Peixuan; LIU Qingjun; ZHANG Zhenlu; LIU Jianying

    2007-01-01

    The histopathological characteristics of congenital aortic valve malformations in children were investigated.All the native surgically excised aortic valves from 32 pediatric patients suffering from symptomatic aortic valve dysfunction due to congenital aortic valve malformations between January 2003 and December 2005 were studied macroscopically and microscopically.The patients' medical records were reviewed and the clinical information was extracted.The diagnosis was made by the clinical presentation,preoperative echocardiography,intraoperative examination,and postoperative histopathological study,excluding rheumatic ot degenerative aortic valve diseases,infective endocarditis and primary connective tissue disorders,e.g.Marfan syndrome.Among 32 children with congenital aortic valve malformations,the age was ranged from six to 18 years,with a mean of 14.9 years,and there were 27 boys and five girls (male:female = 5.4:1).There were five cases of aortic stenosis (AS,15.62%),25 cases of aortic insufficiency (AI,78.13 %)and two cases of AS-AI (6.25%),without other valve diseases.Twenty cases still had other congenital heart diseases:ventricular septal defect (19 cases),patent ductus arteriosus (two cases),double-chambered right ventricle (one case),aneurysm of the right anterior aortic sinus of valsalva (three cases).Histopathological examination indicated that the cusps became thickening with unequal size,irregular shape (coiling and prolapse edge),enhanced hardness,and partly calcification.Microscopic investigation revealed the unsharp structure of valve tissue,fibrosis,myxomatous,reduced collagen fiber,rupture of elastic fibers,different degrees of infiltration of inflammatory cells,secondary calcareous and lipid deposit,and secondary fibrosis.Congenital aortic valve malformations in children involve males more than females,mostly associated with other congenital heart diseases.Aortic insufficiency is more common in children with congenital aortic valve

  6. Morphea simulating paucibacillary leprosy clinically and histopathologically

    Directory of Open Access Journals (Sweden)

    José Saulo Torres Delgado

    2013-01-01

    Full Text Available Clinically and histopathologically paucibacillary leprosy shows similar features with initial morphea. In this case we report a 24 yr-old male patient who presented to our dermatology department with diagnosed paucibacillary leprosy by his local dermatologist, and confirmed by perineurovascular lymphocytic infiltrate in the histopathological exam. On physical examination we found new plaque lesions that were suggestive of morphea with alteration of sensitivity. A new biopsy was performed showing sclerotic superficial dermis with thickening of the collagen bundles in deep dermis and linear arrays lymphocytic infiltrate between the collagen bundles that confirm the diagnosis of morphea.

  7. Mastitis, a Radiographic, Clinical, and Histopathologic Review.

    Science.gov (United States)

    Cheng, Lin; Reddy, Vijaya; Solmos, Gene; Watkins, Latanja; Cimbaluk, David; Bitterman, Pincas; Ghai, Ritu; Gattuso, Paolo

    2015-01-01

    Mastitis is a benign inflammatory process of the breast with heterogeneous histopathological findings, which clinically and radiographically may mimic a mammary carcinoma. We undertook a retrospective study on 37 cases of mastitis in our institution to correlate the radiographic imaging features and the clinical presentation with the histopathological findings. Histologically, there were 21 granulomatous, 7 fibrous, 3 plasma cell, 3 lupus, 2 lymphocytic, and 1 case of acute mastitis. Radiographically, 16/25 (64%) patients with ultrasound studies showed irregular hypoechoic masses suspicious for malignancy. Clinically, 38% of patients had an associated systemic disease. © 2015 Wiley Periodicals, Inc.

  8. Extraction of nucleic acids from yeast cells and plant tissues using ethanol as medium for sample preservation and cell disruption.

    Science.gov (United States)

    Linke, Bettina; Schröder, Kersten; Arter, Juliane; Gasperazzo, Tatiana; Woehlecke, Holger; Ehwald, Rudolf

    2010-09-01

    Here we report that dehydrated ethanol is an excellent medium for both in situ preservation of nucleic acids and cell disruption of plant and yeast cells. Cell disruption was strongly facilitated by prior dehydration of the ethanol using dehydrated zeolite. Following removal of ethanol, nucleic acids were extracted from the homogenate pellet using denaturing buffers. The method provided DNA and RNA of high yield and integrity. Whereas cell wall disruption was essential for extraction of DNA and large RNA molecules, smaller molecules such as tRNAs could be selectively extracted from undisrupted, ethanol-treated yeast cells. Our results demonstrate the utility of absolute ethanol for sample fixation, cell membrane and cell wall disruption, as well as preservation of nucleic acids during sample storage.

  9. Semiempirical Rules To Determine Drug Sensitivity and Ionization Efficiency in Secondary Ion Mass Spectrometry Using a Model Tissue Sample.

    Science.gov (United States)

    Vorng, Jean-Luc; Kotowska, Anna M; Passarelli, Melissa K; West, Andrew; Marshall, Peter S; Havelund, Rasmus; Seah, Martin P; Dollery, Colin T; Rakowska, Paulina D; Gilmore, Ian S

    2016-11-15

    There is an increasing need in the pharmaceutical industry to reduce drug failure at late stage and thus reduce the cost of developing a new medicine. Since most drug targets are intracellular, this requires a better understanding of the drug disposition within a cell. Secondary ion mass spectrometry has been identified as a potentially important technique to do this, as it is label-free and allows imaging in 3D with subcellular resolution and recent studies have shown promise for amiodarone. An important analytical parameter is sensitivity, and we measure this in a bovine liver homogenate reference sample for 20 drugs representing important class types relevant to the pharmaceutical industry. We also measure the sensitivity for pure drug and show, for the first time, that the secondary ion mass spectrometry (SIMS) positive ionization efficiency for small molecules is a simple power-law relationship to the log P value. This discovery will be important for advancing the understanding of the SIMS ionization process in small molecules that has, until now, been elusive. This simple relationship is found to hold true for drug doped in the bovine liver homogenate reference sample, except for fluticasone, nicardipine, and sorafenib which suffer from severe matrix suppression. This relationship provides a simple semiempirical method to determine drug sensitivity for positive secondary ions. Furthermore, we show, on chosen models, how the use of different solvents during sample preparation can affect the ionization of analytes.

  10. Histopathological analysis of unilateral condylar hyperplasia: difficulties in diagnosis and characterization of the disease.

    Science.gov (United States)

    Vásquez, B; Olate, S; Cantín, M; Sandoval, C; Fariña, R; Del Sol, M

    2016-05-01

    The aim of this study was to perform a histological analysis of the conditions in patients undergoing surgery for unilateral condylar hyperplasia (UCH) using different histopathological analytical techniques and to describe the complications and existing controversy in order to better define the disease. A partial condylectomy was performed in five patients who had been diagnosed with UCH using clinical and imaging methods. The samples obtained were analyzed using routine histological techniques including haematoxylin-eosin, Van Gieson, picrosirius, alcian blue/haematoxylin-eosin, and AgNOR staining. The analyses were performed by an observer who was blinded to the clinical parameters of the disease. The cellularity, tissue layers, size of the anatomical structures, and the relationships between them were assessed. The analysis of these patients was complemented by a review of the scientific literature. Variability was observed in the analysis of the cases. The presence of connective tissue was detected at the bone level, with cartilage formation at different levels. Each island presented levels of involvement that could indicate various degrees of aggressiveness. Type I collagen was observed in most cases, although type III was also identified. The development of histological diagnostic methods to determine the aggressiveness or level of involvement in UCH is not currently possible. Further studies are needed to establish new histological classifications.

  11. Drug reaction with eosinophilia and systemic symptoms (DRESS): A histopathology based analysis.

    Science.gov (United States)

    Sasidharanpillai, Sarita; Govindan, Aparna; Riyaz, Najeeba; Binitha, Manikoth P; Muhammed, Kunnummal; Khader, Anza; Reena Mariyath, Olasseri K; Simin, Muhammedkutty; Subin, Kunnari

    2016-01-01

    The data on the histology of cutaneous lesions of drug reaction with eosinophilia and systemic symptoms (DRESS) is limited. To study the histopathology of cutaneous lesions of drug reaction with eosinophilia and systemic symptoms (DRESS) and to identify any features with diagnostic or prognostic significance. All patients admitted to the dermatology ward of government medical college, Kozhikode from January 1, 2014 to December 31, 2014 with probable or definite DRESS as per the RegiSCAR scoring system and who were willing to undergo skin biopsy were included in this prospective study. The study population comprised of nine patients. The consistent histological finding documented was the predominantly lymphocytic dermal inflammatory infiltrate. Four of the five patients whose histology revealed focal interface dermatitis and keratinocyte vacuolation with or without apoptotic keratinocytes, had elevated liver transaminases. Tissue eosinophilia was associated with disease flares. The presence of atypical lymphocytes in peripheral smear and histological evidence of dense dermal inflammatory infiltrate showed an association with hepatic involvement. The main limitations of our study were the small sample size and our inability to carry out a detailed immunohistochemistry work-up. In the appropriate setting, varying combinations of epidermal hyperplasia, spongiosis, parakeratosis and individually necrotic keratinocytes in the background of lymphocyte predominant dermal infiltrate (with some atypia) favor a diagnosis of drug reaction with eosinophilia and systemic symptoms. Female sex, the presence of atypical lymphocytes in peripheral smear, dense dermal inflammatory infiltrate, tissue eosinophilia and interface dermatitis with or without keratinocyt