1.
Suppression of the Epidermal Growth Factor Receptor Inhibits Epithelial-Mesenchymal Transition in Human Pancreatic Cancer PANC-1 Cells.
Source
Department of General Surgery, Peking University First Hospital, 8th Xishiku Street, Xicheng District, Beijing, 100034, People's Republic of China.
Abstract
BACKGROUND:
Aberrant expression of epidermal growth factor receptor (EGFR) has been detected in pancreatic cancer; however, the mechanisms of EGFR in inducing pancreatic cancer development have not been adequately elucidated. The objective of this study was to determine the role of EGFR in mediating epithelial-mesenchymal transition (EMT) in pancreatic cancer cells.
METHODS:
Pancreatic cancer cell line PANC-1 was transfected with small interfering RNA of EGFR by use of a lentiviral expression vector to establish an EGFR-knockdown cell line (si-PANC-1). PANC-1 cells transfected with lentiviral vector expressing negative control sequence were used as negative control (NC-PANC-1). Scratch assay and transwell study were used to analyze cell migration and invasion. Real-time PCR and Western blotting were used to detect the expression of EMT markers E-cadherin, N-cadherin, vimentin, and fibronectin and transcription factors snail, slug, twist1, and sip1 in PANC-1, NC-PANC-1, and si-PANC-1 cells. Immunofluorescent staining with these antibodiesand confocal microscopy were used to observe their cellular location and morphologic changes.
RESULTS:
After RNA interference of EGFR, the migration and invasion ability of si-PANC-1 cells decreased significantly. The expression of epithelial phenotype marker E-cadherin increased and the expression of mesenchymal phenotype markers N-cadherin, vimentin, and fibronectin decreased, indicating reversion of EMT. We also observed intracellular translocation of E-cadherin. Expression of transcription factors snail and slug in si-PANC-1 cells decreased significantly.
CONCLUSION:
Suppression of EGFR expression can significantly inhibit EMT of pancreatic cancer PANC-1 cells. The mechanism may be related with the down-regulation of the expression of transcription factors snail and slug.
Dormant but migratory tumour cells in desmoplastic stroma of invasive ductal carcinomas.
Source
Australian Centre for Microscopy and Microanalysis (ACMM), AMMRF, The University of Sydney, Madsen Building F09, Room 243, Sydney, NSW, 2006, Australia.
Abstract
Mortality in breast cancer is linked to metastasis and recurrence yet there is no acceptable biological model for cancer relapse. We hypothesise that there might exist primary tumour cells capable of escaping surgery by migration and resisting radiotherapy and chemotherapy to cause cancer recurrence. We investigated this possibility in invasive ductal carcinoma (IDC) tissue and observed the presence of solitary primary tumour cells (SPCs) in the dense collagen stroma that encapsulates intratumoural cells (ICs). In IDC tissue sections, collagen was detected with either Masson's Trichrome or by second harmonics imaging. Cytokeratin-19 (CK-19) and vimentin (VIM) antibodies were, respectively, used to identify epithelial-derived tumour cells and to indicate epithelial to mesenchymal transition (EMT). Confocal/multiphoton microscopy showed that ICs from acini were mainly CK-19(+ve) and were encapsulated by dense stromal collagen. Within the stroma, SPCs were detected by their staining for both CK-19 and VIM (confirming EMT). ICs and SPCs were subsequently isolated by laser capture microdissection followed by multiplex tandem-PCR studies. SPCs were found to be enriched for pro-migratory and anti-proliferative genes relative to ICs. In vitro experiments using collagen matrices at 20 mg/cm(3), similar in density to tumour matrices, demonstrated that SPC-like cells were highly migratory but dormant, phenotypes that recapitulated the genotypes of SPCs in clinical tissue. These data suggest that SPCs located at the breast cancer perimeter are invasive and dormant such that they may exceed surgical margins and resist local and adjuvant therapies. This study has important connotations for a role of SPCs in local recurrence.
A distinct microvascular endothelial gene expression profile in severe IUGR placentas.
Source
Research Centre for Women's and Infants' Health, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 25 Orde St, Toronto, Ontario, Canada M5T 3H7.
Abstract
The placental microvasculature is essential for efficient transfer of gases, nutrients and waste between the mother and fetus. Microvascular hypoplasia of the terminal villi is a common pathology in severe Intra Uterine Growth Restriction (IUGR). We used novel methods to obtain placental micro-vascular endothelial cells (PlMEC) from preterm control placentas (n = 3) and placentas from pregnancies with severe IUGR (n = 6) with absent or reversed end-diastolic velocity in the umbilical artery. Distal placental villous tissue was collected to enrich for intermediate and terminal villi. Tissue was digested and PlMEC positively selected using tocosylated magnetic Dynabeads labeled with Human Endothelial Antigen lectin. The purity of the PlMEC (94 ± 2 SD %) was assessed by CD31 and vimentin immunocytochemistry. RNA was extracted from the PlMEC samples and subjected to Affymetrix microarray analysis (U133Plus2 array chips). Comparison of preterm and IUGR PlMEC gene expression profiles identified BTNL9 and NTRK2 transcripts to be upregulated and SAA1 and SLAMF1 transcripts to be downregulated in all 6 IUGR cases relative to preterm controls. A third downregulated gene GNAS was identified to be near significance. Changes were demonstrated to be significant at the mRNA level by Real Time PCR in the PlMEC samples. Changes in the IUGR endothelium were confirmed at the protein level by immunohistochemistry for the 3 with available antibodies. We used a tissue microarray constructed from an independent cohort of placental samples from severe IUGR (n = 7), preeclamptic (n = 7), preterm control (n = 6) and term control (n = 6) pregnancies. Results confirmed differential endothelial expression of BTNL9, NTRK2 and SLAMF1 in IUGR versus preterm and term samples. These studies are the first to characterize PlMEC gene expression profiles thus we have advanced our understanding of the molecular basis of placental micro-vascular pathophysiology in fetal growth restriction.
Crown Copyright © 2012. Published by Elsevier Ltd. All rights reserved.
p63, CK7, PAX8 and INI-1: an optimal immunohistochemical panel to distinguish poorly differentiated urothelial cell carcinoma from high-grade tumours of the renal collecting system.
Source
Department of Pathology, University of Michigan Medical Center, Ann Arbor, MI, USA Division of Urologic Pathology, Caris Life Sciences, Irving, TX, USA.
Abstract
Carvalho J C, Thomas D G, McHugh J B, Shah R B & Kunju L P (2012) Histopathology p63, CK7, PAX8 and INI-1: an optimal immunohistochemical panel to distinguish poorly differentiated urothelial cell carcinoma from high-grade tumours of the renal collecting system Aims: High-grade, poorly differentiated, infiltrative carcinomas involving the renal sinus region often pose challenging differential diagnostic considerations, specifically differentiation of urothelial carcinoma (UC) from renal cell carcinoma (RCC) subtypes. Accurate classification, especially the distinction of UC from RCC, is critical, as therapeutic approaches differ. Methods and results: Cluster analysis was performed on immunohistochemical data from 18 invasive UCs, six CDCs, two RMCs, 18 type 2 papillary renal cell carcinomas (PRCCs) and 20 high-grade clear cell renal cell carcinomas (CRCCs) using a broad panel of traditional and novel immunohistochemical markers. The initial analysis with all antibodies segregates almost all the RCCs (45 of 46, 98%) from all the UCs based on the lack of expression of p63 in all (100%) RCCs, along with predominant strong expression of paired box gene 8 (PAX8) and vimentin, predominant lack of expression of high molecular weight cytokeratin (HMCK) and CK7 and variable expression of RCC, CD10, CA1X and PAX2. All the UCs cluster together with strong, diffuse reactivity for p63, predominant reactivity for CK7 and high molecular weight kininogen (HMWK), and absent to minimal staining with PAX8, RCC antigen, PAX2, alpha-methylacyl-CoA racemase (AMACR), carbonic anhydrase IX (CAIX) andvimentin. After removing antibodies with significant overlap and/or minimal impact, a second analysis with a limited panel including p63, CK7, vimentin, integrase interactor 1 (INI-1) and PAX8 was performed. Again, the majority of UCs cluster into one group and p63 positivity separates all UCs from RCCs. Conclusions: Lack of INI-1 expression, noted exclusively in RMCs, segregates RMCs into a separate cluster. PAX8 is rarely positive (17%) in UC, is commonly expressed in CDC, RMC, PRCC and CRCC and is superior to PAX2.
© 2012 Blackwell Publishing Ltd.
Molecular characteristics and alterations during early development of the human vagina.
Source
Division of Clinical and Functional Anatomy, Department of Anatomy, Histology and Embryology, Innsbruck Medical University, Innsbruck, Austria.
Abstract
Unresolved questions remain concerning the derivation of the vagina with respect to the relative contributions from the Müllerian ducts, the urogenital sinus, and the Wolffian ducts. Recent molecular and cellular studies in rodents have opened up a large gap between the level of understanding of vaginal development in mice and understanding of human vaginal development, which is based on histology. To compare the findings in mice with human vaginal development and to address this gap, we analysed molecular characteristics of the urogenital sinus, Wolffian ducts, and Müllerian ducts in 8-14-week-old human specimens using immunohistochemical methods. The monoclonal antibodies used were directed against cytokeratin (CK) 14, CK19, vimentin, laminin, p63, E-cadherin, caspase-3, Ki67, HOX A13, and BMP-4. The immunohistochemical analysis revealed that, during weeks 8-9, the epithelium of the Müllerian ducts became positive for p63 as p63-positive cells that originated from the sinus epithelium reached the caudal tip of the fused Müllerian ducts via the Wolffian ducts. The lumen of the fused Müllerian ducts was closed by an epithelial plug that contained both vimentin-positive and vimentin-negative cells. Subsequently, the resulting epithelial tube enlarged by proliferation of basal p63-positive cells. The first signs of squamous differentiation were detected during week 14, with the appearance of CK14-positive cells. According to our results, all three components, namely, the urogenital sinus, Wolffian ducts, and Müllerian ducts, interacted during the formation of the human vagina. The sinus epithelium provided p63-positive cells, the Wollfian ducts acted as a 'transporter', and the Müllerian ducts contributed the guiding structure for the vaginal anlagen. Epithelial differentiation began at the end of the period studied and extended in a caudo-cranial direction. The present study is one of the first to provide up-to-date molecular correlates for human vaginal development that can be compared with the results of cell biological studies in rodents.
© 2012 The Authors. Journal of Anatomy © 2012 Anatomical Society.
Immunocytochemical Localization of Cytoplasmic and Nuclear Intermediate Filaments in the Bovine Ovary during Folliculogenesis.
Source
Lehrstuhl für Anatomie, Histologie und Embryologie, Department of Veterinary Sciences, LMU München, Veterinärstrasse 13, D-80539, Munich, Germany.
Abstract
The cellular cytoskeleton is composed of three fibrillar systems, namely actin microfilaments, microtubules and intermediate filaments (IFs). It not only is a structural system, which mediates functional compartmentalization, but also contributes to many cellular processes such as transport, mitosis, secretion, formation of cell extensions, intercellular communication and apoptosis. In this study, we have examined the distribution of four groups of IFs [cytokeratins (CKs),vimentin, desmin and lamins] in the somatic and germinal cells of the bovine ovary using RT-PCR and immunohistochemical techniques. Using RT-PCR, specific transcripts for all intermediate proteins studied (CK8, CK18, desmin, vimentin, lamin A/C and lamin B1) were detected. A characteristic immunohistochemical staining pattern was observed for the different IFs within the ovary. In this study, we used antibodies against type I CK (acidic CKs: CK14, CK18 and CK19) and type II CK (basic CKs: CK5 and CK8). Among these, only antibodies against CK18 gave a characteristic pattern of immunostaining in the ovary, which included the surface epithelium, the follicle cells, the endothelium of blood vessels and rete ovarii. Antibodies against all other CKs resulted in a weak staining of a limited number of cellular structures (CK5 and CK19) or were completely negative (CK8 and CK14, apart from the surface epithelium). Vimentin antibodies resulted occasionally in a weak staining of the granulosa cells of primary and secondary follicles. In late secondary follicles, the basal and the most apical follicle cells contacting the zona pellucida usually showed a marked immunostaining for vimentin. In antral follicles, three different immunostaining patterns forvimentin were observed. Desmin immunostaining was confined to the smooth muscle cells of blood vessels. Although mRNA for lamin A/C and lamin B1 could be demonstrated using RT-PCR, no immunostaining was found for lamins, neither in the follicle cells nor in the oocytes.
© 2012 Blackwell Verlag GmbH.
Hepatoma-Derived Growth Factor Regulates breast cancer cell invasion by Modulating Epithelial Mesenchymal Transition.
Source
Institute of Biomedical Sciences, National Sun Yat-Sen University, Kaohsiung 804, Taiwan; Center for Neuroscience, National Sun Yat-Sen University, Kaohsiung 804, Taiwan.
Abstract
Hepatoma-derived growth factor (HDGF) participates in tumorigenesis but its role in breast cancer is unclear. We set out to elucidate the expression profile and function of HDGF during breast carcinogenesis. Immunoblot and immunohistochemical studies revealed elevated HDGF expression in human breast cancer cell lines and tissues. Nuclear HDGF labeling index was positively correlated with tumour grade, stage, and proliferation index, but negatively correlated with survival rate in breast cancer patients. HDGF overexpression was associated with lymph node metastasis and represented an independent prognostic factor for tumour recurrence. Gene transfer studies were performed to elucidate the influence of cellular HDGF level on the malignant behaviour and epithelial-mesenchymal transition (EMT) of breast cancer cells. Adenovirus-mediated HDGF overexpression stimulated the invasiveness and colony formation of MCF-7 cells. Moreover, HDGF overexpression promoted breast cancer cell EMT by E-cadherin downregulation and vimentin upregulation. Conversely, HDGF knockdown by RNA interference in MDA-MB-231 cells attenuated the malignant behaviour and elicited EMT reversal by enhancing E-cadherin expression while depletingvimentin expression. Because HDGF is a secreted protein, we evaluated the cellular function of recombinant HDGF and found that exogenously supplied HDGF enhanced the invasiveness of breast cancer cells by down-regulating E-cadherin and up-regulating vimentin at transcriptional and translational levels. In contrast, blockade of HDGF secretion with an HDGF antibody inhibited the malignant behaviours and EMT. Finally, exogenous HDGF partially reversed benzyl isothiocyanate (BITC)-induced EMT suppression. HDGF overexpression may exert a prognostic role for tumour metastasis and recurrence in breast cancer by modulating EMT. Copyright © 2012 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Copyright © 2012 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
ZEB2 upregulates integrin α5 expression through cooperation with Sp1 to induce invasion during epithelial-mesenchymal transition of human cancer cells.
Source
Therapeutic Antibody Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejon, Korea.
Abstract
Epithelial-mesenchymal transition (EMT) is a process implicated in tumor invasion, metastasis, embryonic development, and wound healing. ZEB2 is a transcription factor involved in EMT that represses E-cadherin transcription. Although E-cadherin downregulation is a major event during EMT and tumor progression, E-cadherin reduction is probably not sufficient for full invasiveness. The mechanisms by which E-cadherin transcriptional repressors induce mesenchymal genes during EMT remain largely unknown. Here, we investigated the role of ZEB2 in the induction of integrin α5 during cancer EMT and its underlying mechanism. In human cancer cells, ZEB2 was found to directly upregulate integrin α5 transcription in a manner that is independent of the regulation of E-cadherin expression. Conversely, depletion of ZEB2 by small interfering RNA suppressed integrin α5 expression, leading to reduced invasion. Suppression of integrin α5 inhibited cancer cell invasion, suggesting an important role for integrin α5 in cancer progression. Furthermore, ZEB2 was found to activate the integrin α5 and vimentin promoters by interacting with and activating the transcription factor Sp1, suggesting that cooperation between ZEB2 and Sp1 represents a novel mechanism of mesenchymal gene activation during EMT. These findings increase our understanding of the pathways beyond E-cadherin reduction that regulate mesenchymal gene expression during EMT and cancer progression.
Pregnancy-Related Peritoneal Ectopic Decidua (Deciduosis): Morphological and Clinical Evaluation.
Source
Department of Pathology, Başkent University, Faculty of Medicine, ANKARA, TURKEY.
Abstract
Objective: Ectopic decidual reaction (deciduosis) can be seen rarely on the peritoneum during laparotomy for a cesarean section for pregnancy, in addition to the ovary and cervix. The aim of this study was to evaluate the clinical, histopathological, immunohistochemical characteristics of ectopic decidua cases that were incidentally found in the peritoneum during a cesarean section. Material and Method: A total of seven cases where decidualization was found in the peritoneal biopsy taken during pregnancy at the Baskent University Medical Faculty Adana Teaching and Training Hospital Department of Pathology were included in this study. The clinical features of the cases were obtained from their clinical folders. The morphological findings were recorded and the peritoneal biopsies were analyzed with keratin Pan Ab-1, calretinin, vimentin, HMB-45 and progesterone receptor antibody for immunohistochemical analysis. Results: The mean age for the seven cases was 36±4.16. The gestational age was 33 to 39 (mean 37.2) weeks. Microscopic evaluation revealed decidualized cells that were large polygonal and eosinophilic, some with vacuolated cytoplasm, that formed small nodules under the mesothelium of the peritoneum in all cases. Immunohistochemical staining showed positive staining of the cell cytoplasm with vimentin and positive staining of the cell nucleus with the progesterone receptor antibody in the decidual cells. Calretinin, keratin and HMB-45 stains were negative. Conclusion: Pregnancy-related peritoneal deciduosis develops with the effect of progesterone in pregnancy. It disappears without complication in the postpartum period. Immunohistochemistry may help the differential diagnosis of peritoneal deciduosis where problems are experienced differentiating the case from malignant mesothelioma or metastatic tumor.
Clinical and histopathological profile of primary or secondary osteosarcoma of the jaws.
Source
Ospedale San Martino-Padiglione 4, Largo Rosanna Benzi, 10 (16132) Genova, Italy. f.angiero@teos.it.
Abstract
Osteosarcoma of the jaw is a rare disease; we report two cases, one in which the primary osteosarcoma had occurred in the sacrum and ileum, the second at the mandible. Dissemination of osteosarcoma to other organs, especially early dissemination to the lung, is common, but metastasis to the jaw has only rarely been reported. About 10% of osteosarcomas occur in the head and neck, most in the mandible or maxilla. Clinically, both patients presented swelling, and pain at the jaw in the premolar-molar region. At radiography, extensive bone erosion and soft-tissue swelling were apparent. A biopsy was taken and a diagnosis of osteosarcoma rendered in both cases. Histological examination revealed a proliferation of atypical osteoblast-like cells with hyperchromatic nuclei and formation of scattered neoplastic osteoid tissue. Immunohistochemistry for a panel of antibodies showed strong positivity for CD99, weak positivity for S-100, but was negative for desmin, vimentin, and cytokeratins. The diagnosis for both cases was of osteogenic osteosarcoma, chondroblastic subtype. Unfortunately, both patients died, one before the planned chemotherapy regime could begin, the second during the chemotherapy course. Our report aims to highlight the importance of the diagnostic profile in formulating a diagnosis of osteosarcoma, and that this tumor, although very rare, may be primary or may metastasize to the jaws.
- PMID:
- 22199320
- [PubMed - in process]
Spontaneous Nephroblastoma in a Japanese Giant Salamander (Andrias japonicus).
Source
Laboratory of Veterinary Pathology, Azabu University.
Abstract
An adult male Japanese giant salamander (Andrias japonicus) died accidentally, and necropsy showed a white mass (23 × 15 mm) in the left kidney and hepatorrhexis with hemoperitoneum. Histologically, the renal mass was mainly composed of immature nephroblastic tumor cells. In the tumor tissue, a trabecular pattern lined by oval to polygonal tumor cells with rich interstitium, solid growth, and a few tubular structures was observed. Nephroblastic tumor cells were weakly positive and epithelium-like tumor cells were strongly positive for cytokeratin. However, antibodies forvimentin and Wilms' tumor protein 1 did not react with the salamander's cells. On electron microscopy, a desmosome junction was observed between tumor cells. This is the first report of nephroblastoma in a Japanese giant salamander.
The presence of anti-citrullinated protein antibodies (ACPA) and rheumatoid factor on patients with rheumatoid arthritis (RA) does not interfere with the chance of clinical remission in a follow-up of 3 years.
Source
Rheumatology Division, Hospital Universitário de Brasília da Universidade de Brasília, SHLS 716/916 Bloco E salas 501-502, Centro Médico de Brasília, Asa Sul, Brasilia, DF, CEP: 71660020, Brazil, licia@unb.br.
Abstract
Autoantibodies in early rheumatoid arthritis (RA) have important diagnostic value. The association between the presence of autoantibodies against cyclic citrullinated peptide and the response to treatment is controversial. To prospectively evaluate a cohort of patients with early rheumatoid arthritis (<12 months of symptoms) in order to determine the association between serological markers (rheumatoid factor (RF), anti-citrullinated protein antibodies) such as anti-cyclic citrullinated peptide antibodies (anti-CCP) and citrullinated anti-vimentin (anti-Sa) with the occurrence of clinical remission, forty patients diagnosed with early RA at the time of diagnosis were evaluated and followed for 3 years, in use of standardized therapeutic treatment. Demographic and clinical data were recorded, disease activity score 28 (DAS 28), as well as serology tests (ELISA) for RF (IgM, IgG, and IgA), anti-CCP (CCP2, CCP3, and CCP3.1) and anti-Sa in the initial evaluation and at 3, 6, 12, 18, 24, and 36 months of follow-up. The outcome evaluated was the percentage of patients with clinical remission, which was defined by DAS 28 lower than 2.6. Comparisons were made through the Student t test, mixed-effects regression analysis, and analysis of variance (significance level of 5%). The mean age was 45 years, and a female predominance was observed (90%). At the time of diagnosis, RF was observed in 50% of cases (RF IgA-42%, RF IgG-30%, and RF IgM-50%), anti-CCP in 50% (no difference between CCP2, CCP3, and CCP3.1) and anti-Sa in 10%. After 3 years, no change in the RF prevalence and anti-CCP was observed, but the anti-Sa increased to 17.5% (P = 0.001). The percentage of patients in remission, low, moderate, and intense disease activity, according to the DAS 28, was of 0, 0, 7.5, and 92.5% (initial evaluation) and 22.5, 7.5, 32.5, and 37.5% (after 3 years). There were no associations of the presence of autoantibodies in baseline evaluation and in serial analysis with the percentage of clinical remission during follow-up of 3 years The presence of autoantibodies in early RA has no predictive value for clinical remission in early RA.
Involvement of epithelial mesenchymal transition of human endometrial epithelial cells in human embryo implantation: evidence from studies using in vitro model.
Source
Keio University School of Medicine, Japan.
Abstract
Human embryo implantation is a critical multistep process consisting of embryo apposition/adhesion, followed by penetration and invasion. Through embryo penetration, the endometrial epithelial cell barrier is disrupted and remodeled by an unknown mechanism. We have previously developed an in vitro model for human embryo implantation employing the human choriocarcinoma cell line JAR and the human endometrial adenocarcinoma cell line Ishikawa. Using this model we have shown that stimulation with ovarian steroid hormones (17β-estradiol and progesterone, E2P4) and suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, enhances the attachment and adhesion of JAR spheroids to Ishikawa. In the present study we showed that the attachment and adhesion of JAR spheroids and treatment with E2P4 or SAHA individually induce the epithelial mesenchymal transition (EMT) in Ishikawa cells. This was evident by up-regulation of N-cadherin and vimentin, a mesenchymal cell marker, and concomitant down-regulation of E-cadherin in Ishikawa cells. Stimulation with E2P4 or SAHA accelerated Ishikawa cell motility, increased JAR spheroid outgrowth, and enhanced the unique redistribution of N-cadherin, which was most prominent in proximity to the adhered spheroids. Moreover, an N-cadherin functional blocking antibody attenuated all events but not JAR spheroid adhesion. These results collectively provide evidence suggesting that E2P4- and implanting embryo-induced EMT of EECs may play a pivotal role in the subsequent processes of human embryo implantation with functional control of N-cadherin.
Intravascular papillary endothelial hyperplasia of the oral cavity.
Source
Department of Diagnostic and Therapeutic Sciences, Meikai University School of Dentistry, Sakado, Japan.
Abstract
To examine the pathogenesis of intravascular papillary endothelial hyperplasia (IPEH), a relatively uncommon benign, non-neoplastic vascular lesion, clinicopathological and immunohistochemical studies were performed. Paraffin-embedded tissue specimens of 78 vascular lesions were examined histologically, and 9 cases of IPEH were investigated immunohistochemically using antibodies against CD34, vimentin, factor VIII antigen, α-smooth muscle actin (α-SMA), podoplanin, CD105, and ki-67 antigen. A thrombus or ulcer was found near the sites of all IPEH specimens. Histologic examination revealed papillary proliferated endothelial cells located toward the lumen of enlarged blood vessels. Immunohistochemistry showed that CD34, α-SMA, and factor VIII antigen were positive in lining endothelial cells. Vimentin was positive in the mesenchymal components. Immunohistochemical staining for podoplanin and CD105 was partially positive. Labeling index was 4.7 to 9.2 in ki-67-positive cases. IPEH is believed to result from reactive proliferation of blood endothelial cells that is caused by an abnormal process of organization in thrombosed blood vessels. The pathogenesis of IPEH might be related to inflammation or mechanical stimulus such as irritation.
Immunohistochemical Characteristics of Normal Canine Eyes.
Abstract
Immunohistochemistry is widely utilized in diagnostic laboratories to study neoplastic and nonneoplastic diseases. Knowledge of the immunohistochemical characteristics of normal tissue is essential for interpretation of immunoreactivity in pathologic conditions. In this study, immunohistochemistry was performed with a broad panel of diagnostically relevant antibodies on 4 normal canine globes-namely, vimentin, pan-cytokeratin (AE1/AE3), cytokeratin 7, cytokeratin 8/18, cytokeratin 20, α-smooth muscle actin, muscle specific actin, desmin, Melan-A, microphthalmia transcription factor, S-100, glial fibrillary acidic protein, triple neurofilaments, neuron-specific enolase, chromogranin A, synaptophysin, laminin and CD31. Results include cytokeratin immunoreactivity limited to the conjunctival epithelium, corneal epithelium, and retinal pigment epithelium; distinct patterns of immunopositivity of muscle markers; and widespread immunoreactivity for vimentin and most neural/neuroendocrine markers. These findings in normal eyes provide the basis for interpretation of ocular immunohistochemistry in dogs. Published immunophenotypes of primary ocular neoplasms are also reviewed.
- PMID:
- 22156227
- [PubMed - as supplied by publisher]
Synchronized expressions of hepatic stellate cells and their transactivation and liver regeneration during liver injury in an animal model of cholestasis.
Source
Department of Pediatric Surgery, Reproductive and Developmental Medicine, Kyushu University, Fukuoka, Japan. safira@pedsurg.med.kyushu-u.ac.jp
Abstract
BACKGROUND:
There is much known about hepatic stellate cells (HSCs) during liver injury. However, some aspects remain unclear, such as the natural expression levels of HSCs during the days to weeks after liver injury. Does liver regeneration start the same time as the injury process?
METHODS:
Fifty-four male Wistar rats aged 7 to 8 weeks, weighing 200 to 320 g each were subjected to bile duct ligation (BDL). After surgery, they were killed at different times post-BDL. Collagen deposition was analyzed, and immunohistochemical staining of α-smooth muscle actin (α-SMA), vimentin, matrix metalloproteinase-2 (MMP-2), tissue inhibitor matrix metalloproteinase-1, and proliferating cell nuclear antigen antibody (PCNA) was performed to evaluate HSCs and liver regeneration.
RESULTS:
The expression of α-SMA was seen as early as day 3 post-BDL, which started from peribiliary to perisinusoidal, and was seen throughout the whole liver sections on day 28 post-BDL. Similar expression patterns were seen in MMP-2 staining. The PCNA expression was strongest around the perisinusoidal area. These expression patterns were not observed in the sham-operated rats.
CONCLUSIONS:
The activation of HSCs showed a synchronized fibrogenic process and liver regeneration from days to weeks after liver injury. Matrix degradation was thus found to increase in accordance with chronic liver injury, which thus led to an excessive collagen deposition.
Copyright © 2011 Elsevier Inc. All rights reserved.
The diagnostic value of EMA expression in the renal parenchyma tumors.
Source
Department of Histology, Victor Babeş University of Medicine and Pharmacy, Timisoara, Romania.
Abstract
Renal parenchyma tumors are a heterogeneous group of malignancies that are difficult to diagnose and classify. Immunohistochemistry begun to be routinely used for the diagnosis of these tumors. Panels of antibodies are developed for the diagnostic assessment of these tumors, which include cytokeratins, epithelial membrane antigen and vimentin. Epithelial membrane antigen (EMA) is expressed by most of the tumor cell types. Forty-seven specimens of renal parenchyma tumors were studied immunohistochemically for the expression of EMA. In the majority of the cases, clear cells carcinoma was positive for EMA (25/33, 75.70%). All of the papillary carcinomas were positive, with different staining patterns between the two subtypes. The two cases of chromophobe cells carcinomas were intensely positive with a granular cytoplasmic staining pattern. The mixed epithelial-stromal tumor was negative for EMA in both of the components. Out of the three cases of sarcomatoid carcinomas, one was negative, one was weakly positive (+1) and the last was positive (+2). Intensely positive normal tubes were caught by the tumor proliferation in the negative case and in the negative stained areas of the weakly positive case.
Tumor necrosis factor blockade differentially affects innate inflammatory and th17 cytokines in rheumatoid arthritis.
Source
Center for Molecular Medicine and Stem Cell Research, Medical Faculty, University of Kragujevac, Svetozara Markovica 69, 34000 Kragujevac, Serbia. nadap@ikomline.net.
Abstract
OBJECTIVE:
To evaluate the effect of a tumor necrosis factor-α (TNF-α) inhibitor (etanercept) on innate inflammatory and Th17 cytokines in patients with rheumatoid arthritis (RA).
METHODS:
Serum samples were collected from 40 patients with active RA refractory to conventional disease-modifying antirheumatic drugs who initiated therapy with etanercept plus methotrexate (MTX). Treatment response was assessed at Week 24 according to the European League Against Rheumatism response criteria. Serum levels of interleukin 6 (IL-6), TNF-α, IL-32, IL-23, IL-17A, IL-21, and IL-22 were measured in patients with RA and 25 healthy controls.
RESULTS:
Patients with RA had increased levels of IL-6 (p < 0.001), IL-32 (p < 0.001), IL-23 (p < 0.001), and a trend toward increased IL-21 in the sera compared to controls. At 24 weeks' posttreatment, followup serum samples of etanercept responders had decreased levels of IL-6 (p < 0.001) and increased IL-21 (p < 0.05) and IL-32 (p < 0.001), while there were no differences in cytokine levels in non-responders. Serum IL-6 levels were positively correlated with levels of erythrocyte sedimentation rate (r = 0.458, p < 0.01), C-reactive protein (r = 0.593, p < 0.01), and 28-joint Disease Activity Score (r = 0.432, p < 0.01) at baseline. Serum IL-21 levels were positively correlated with levels of rheumatoid factor (r = 0.513, r = 0.633, both p < 0.01) and antimutated citrullinated vimentin antibodies (r = 0.515, p < 0.01; r = 0.428, p < 0.05) at baseline and after 24 weeks of treatment with etanercept.
CONCLUSION:
Multiple inflammatory pathways contribute to persistent chronic inflammation in RA. In contrast to nonresponders, etanercept therapy modulated serum cytokine levels and caused a marked decrease of IL-6 levels in responders. IL-21 might be involved in the regulation of autoantibody production in RA.
The fine specificity of IgM anti-citrullinated protein antibodies (ACPA) is different from that of IgG ACPA.
Source
Department of Rheumatology, Leiden University Medical Center (LUMC), PO Box 9600, NL-2300 RC Leiden, The Netherlands. L.A.Trouw@lumc.nl.
Abstract
ABSTRACT:
INTRODUCTION:
The antigen recognition pattern of immunoglobulin M (IgM) could, when directed against protein antigens, provide an indication of the antigenic moieties triggering new B cells. The half-life of IgM is short and memory B cells against T-cell-dependent protein antigens typically produce IgG and not IgM antibodies. In this study, we analyzed whether a difference exists between the fine specificity of IgM versus IgG anti-citrullinated protein antibodies(ACPAs).
METHODS:
We determined the fine specificity of IgM and IgG ACPAs in 113 ACPA-positive rheumatoid arthritis patients with IgM cyclic citrullinated peptide 2 (CCP2) levels above 100 AU/ml. Fine specificity was assessed by performing ELISA using citrullinated peptides derived from vimentin, fibrinogen-β, fibrinogen-α and α-enolase, as well as citrullinated proteins fibrinogen and myelin basic protein. The arginine counterparts were used as controls.
RESULTS:
Recognition of defined citrullinated antigens by IgM ACPA was confined to samples that also displayed recognition by IgG ACPA. However, the IgM ACPA response displayed a more restricted antigen recognition profile than IgG ACPA (OR = 0.26, P < 0.0001).
CONCLUSION:
Our data show that several defined citrullinated antigens are recognized only by IgG ACPA, whereas others are also recognized by IgM ACPA. These observations suggest that not all citrullinated antigens are able to activate new B cells despite concurrent recognition by IgG ACPA.
Autoantibodies in early rheumatoid arthritis: Brasília cohort: results of a three-year serial analysis.
Source
Serviço de Reumatologia Clínica Médica, Universidade de Brasília. liciamhmota@yahoo.com.br
Abstract
The diagnostic and prognostic value of the serial measurement of antibodies, such as rheumatoid factor (RF), anti-cyclic citrullinated peptide (anti-CCP), and anti-citrullinated vimentin (anti-Sa) antibodies, has not been defined in early rheumatoid arthritis (ERA).
OBJECTIVES:
To prospectively assess the presence of RF, anti-CCP, and anti-Sa in ERA patients.
PATIENTS AND METHODS:
Forty ERA (less than 12 months) patients of the Brasília cohort were evaluated and followed up for three years. Both clinical and demographic data were recorded, in addition to the results (ELISA) of RF (IgM, IgG, and IgA), anti-CCP (CCP2, CCP3, and CCP3.1), and anti-Sa at the baseline assessment and after 3, 6, 12, 18, 24 and 36 months of follow-up. The results were compared by use of Student t test and paired t test.
RESULTS:
The patients' mean age was 45 years, and 90% of them were female. At the time of diagnosis, RF was identified in 50% of the patients (RF IgA, 42%; RF IgG, 30%; and RF IgM, 50%), anti-CCP in 52.5% (no difference between CCP2, CCP3, and CCP3.1), and anti-Sa in 10%. After three years, no difference was observed in RF and anti-CCP prevalence, but anti-Sa increased to 17.5% (P = 0.001).
CONCLUSION:
Repeated RF and anti-CCP measurement, including different isotypes, during three years of follow-up showed no significant changes. The third generation of anti-CCP assays did not increase the diagnostic value of the second-generation assays.
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