ABSTRACT
Brain death (BD) has been associated with an immunological priming of donor organs and is thought to exacerbate ischemia reperfusion injury (IRI). Recently, we showed that the essential nitric oxide synthase co-factor tetrahydrobiopterin (BH4) abrogates IRI following experimental pancreas transplantation. We therefore studied the effects of BD in a murine model of syngeneic pancreas transplantation and tested the therapeutic potential of BH4 treatment. Compared with sham-operated controls, donor BD resulted in intragraft inflammation reflected by induced IL-1ß, IL-6, VCAM-1, and P-selectin mRNA expression levels and impaired microcirculation after reperfusion (p < 0.05), whereas pretreatment of the BD donor with BH4 significantly improved microcirculation after reperfusion (p < 0.05). Moreover, BD had a devastating impact on cell viability, whereas BH4-treated grafts showed a significantly higher percentage of viable cells (p < 0.001). Early parenchymal damage in pancreatic grafts was significantly more pronounced in organs from BD donors than from sham or non-BD donors (p < 0.05), but BH4 pretreatment significantly ameliorated necrotic lesions in BD organs (p < 0.05). Pretreatment of the BD donor with BH4 resulted in significant recipient survival (p < 0.05). Our data provide novel insights into the impact of BD on pancreatic isografts, further demonstrating the potential of donor pretreatment strategies including BH4 for preventing BD-associated injury after transplantation.
Subject(s)
Biopterins/analogs & derivatives , Brain Death/pathology , Pancreas Transplantation/methods , Pancreatitis/pathology , Reperfusion Injury/prevention & control , Analysis of Variance , Animals , Biopterins/pharmacology , Disease Models, Animal , Graft Rejection/prevention & control , Graft Survival , Inflammation Mediators/metabolism , Kaplan-Meier Estimate , Male , Mice , Mice, Inbred C57BL , Microcirculation , Pancreas Transplantation/adverse effects , Pancreatitis/physiopathology , Postoperative Complications/pathology , Random AllocationABSTRACT
Normothermic machine perfusion (NMP) has emerged as an innovative organ preservation technique. Developing an understanding for the donor organ immune cell composition and its dynamic changes during NMP is essential. We aimed for a comprehensive characterization of immune cell (sub)populations, cell trafficking and cytokine release during liver NMP. Single-cell transcriptome profiling of human donor livers prior to, during NMP and after transplantation shows an abundance of CXC chemokine receptor 1+/2+ (CXCR1+/CXCR2+) neutrophils, which significantly decreased during NMP. This is paralleled by a large efflux of passenger leukocytes with neutrophil predominance in the perfusate. During NMP, neutrophils shift from a pro-inflammatory state towards an aged/chronically activated/exhausted phenotype, while anti-inflammatory/tolerogenic monocytes/macrophages are increased. We herein describe the dynamics of the immune cell repertoire, phenotypic immune cell shifts and a dominance of neutrophils during liver NMP, which potentially contribute to the inflammatory response. Our findings may serve as resource to initiate future immune-interventional studies.
Subject(s)
Liver Transplantation , Humans , Aged , Liver Transplantation/methods , Liver , Perfusion/methods , Organ Preservation/methods , Sequence Analysis, RNAABSTRACT
Depletion of the nitric oxide synthase cofactor tetrahydrobiopterin (H4B) during ischemia and reperfusion is associated with severe graft pancreatitis. Since clinically feasible approaches to prevent ischemia reperfusion injury (IRI) by H4B-substitution are missing we investigated its therapeutic potential in a murine pancreas transplantation model using different treatment regimens. Grafts were subjected to 16 h cold ischemia time (CIT) and different treatment regimens: no treatment, 160 µM H4B to perfusion solution, H4B 50 mg/kg prior to reperfusion and H4B 50 mg/kg before recovery of organs. Nontransplanted animals served as controls. Recipient survival and endocrine graft function were assessed. Graft microcirculation was analyzed 2 h after reperfusion by intravital fluorescence microscopy. Parenchymal damage was assessed by histology and nitrotyrosine immunohistochemistry, H4B tissue levels by high pressure liquid chromatography (HPLC). Compared to nontransplanted controls prolonged CIT resulted in significant microcirculatory deterioration. Different efficacy according to route and timing of administration could be observed. Only donor pretreatment with H4B resulted in almost completely abrogated IRI-related damage showing graft microcirculation comparable to nontransplanted controls and restored intragraft H4B levels, resulting in significant reduction of parenchymal damage (p < 0.002) and improved survival and endocrine function (p = 0.0002 each). H4B donor pretreatment abrogates ischemia-induced parenchymal damage and represents a promising strategy to prevent IRI following pancreas transplantation.
Subject(s)
Biopterins/analogs & derivatives , Pancreas Transplantation/methods , Reperfusion Injury/prevention & control , Tissue Donors , Animals , Biopterins/therapeutic use , Cold Ischemia , Male , Mice , Mice, Inbred C57BL , Microcirculation , Models, Animal , Pancreas/blood supply , Pancreas/pathology , Pancreas Transplantation/physiology , Peroxynitrous Acid/biosynthesis , Transplantation, Isogeneic , Tyrosine/analogs & derivatives , Tyrosine/biosynthesisABSTRACT
BACKGROUND: High-level expression of epithelial cell adhesion molecule (EpCAM) is associated with unfavorable prognosis in breast cancer. This study was designed to investigate two doses of the fully human IgG1 anti-EpCAM antibody adecatumumab (MT201) in patients with metastatic breast cancer (MBC). METHODS: A total of 109 patients were stratified into high- and low-level EpCAM expression by immunohistochemical staining of primary tumors and subsequently randomly assigned to receive monotherapy with either high- (6 mg/kg every two weeks (q2w)) or low-dose adecatumumab (2 mg/kg/ q2w) until disease progression. RESULTS: No complete or partial tumor responses could be confirmed by central RECIST assessment. The probability for tumor progression was significantly lower in patients receiving high-dose adecatumumab and expressing high levels of EpCAM (hazard ratio 0.43; P = 0.0057 versus low dose and low EpCAM). Three of 18 patients with highest EpCAM expression treated with adecatumumab developed new metastases up to week 6, compared with 14 of 29 patients with low EpCAM. Most frequent treatment-related adverse events (high dose/low dose) were chills (59%/20%), nausea (55%/18%), fatigue (39%/23%) and diarrhea (43%/7%). CONCLUSIONS: Single-agent adecatumumab shows dose- and target-dependent clinical activity in EpCAM-positive MBC, albeit no objective tumor regression. Further investigation of adecatumumab in patients with EpCAM-overexpressing tumors and lower tumor burden is warranted.
Subject(s)
Antibodies, Monoclonal/therapeutic use , Antigens, Neoplasm/immunology , Breast Neoplasms/drug therapy , Cell Adhesion Molecules/immunology , Cell Adhesion Molecules/therapeutic use , Adult , Aged , Aged, 80 and over , Antibodies, Monoclonal/administration & dosage , Antibodies, Monoclonal/adverse effects , Antibodies, Monoclonal, Humanized , Antineoplastic Agents/administration & dosage , Antineoplastic Agents/adverse effects , Antineoplastic Agents/therapeutic use , Breast Neoplasms/pathology , Cell Adhesion Molecules/administration & dosage , Cell Adhesion Molecules/adverse effects , Disease Progression , Disease-Free Survival , Dose-Response Relationship, Drug , Epithelial Cell Adhesion Molecule , Female , Humans , Middle Aged , Neoplasm Metastasis , Treatment OutcomeABSTRACT
Exposure to competitive mental tasks significantly reduced the urinary sodium and fluid excreted by young men with one or two hypertensive parents or with borderline hypertension. In this high-risk group, the degree of retention was directly related to the magnitude of heart rate increase during stress, suggesting common mediation by way of the sympathetic nervous system. Thus, psychological stress appears to induce changes in renal excretory functions that may play a critical role in long-term blood pressure regulation.
Subject(s)
Hypertension/physiopathology , Sodium/metabolism , Stress, Psychological/physiopathology , Water-Electrolyte Balance , Adolescent , Adult , Blood Pressure , Heart Rate , Humans , Hypertension/etiology , Kidney/physiopathology , Male , Risk , Stress, Psychological/metabolismABSTRACT
Epithelial to mesenchymal transition (EMT) is implicated in the progression of primary tumours towards metastasis and is likely caused by a pathological activation of transcription factors regulating EMT in embryonic development. To analyse EMT-causing pathways in tumourigenesis, we identified transcriptional targets of the E-cadherin repressor ZEB1 in invasive human cancer cells. We show that ZEB1 repressed multiple key determinants of epithelial differentiation and cell-cell adhesion, including the cell polarity genes Crumbs3, HUGL2 and Pals1-associated tight junction protein. ZEB1 associated with their endogenous promoters in vivo, and strongly repressed promotor activities in reporter assays. ZEB1 downregulation in undifferentiated cancer cells by RNA interference was sufficient to upregulate expression of these cell polarity genes on the RNA and protein level, to re-establish epithelial features and to impair cell motility in vitro. In human colorectal cancer, ZEB1 expression was limited to the tumour-host interface and was accompanied by loss of intercellular adhesion and tumour cell invasion. In invasive ductal and lobular breast cancer, upregulation of ZEB1 was stringently coupled to cancer cell dedifferentiation. Our data show that ZEB1 represents a key player in pathologic EMTs associated with tumour progression.
Subject(s)
Breast Neoplasms/pathology , Cell Differentiation , Cell Polarity , Colonic Neoplasms/pathology , Cytoskeletal Proteins/antagonists & inhibitors , Homeodomain Proteins/metabolism , Membrane Glycoproteins/antagonists & inhibitors , Membrane Proteins/antagonists & inhibitors , Nucleoside-Phosphate Kinase/antagonists & inhibitors , Transcription Factors/metabolism , Adult , Aged , Breast Neoplasms/genetics , Breast Neoplasms/metabolism , Cadherins/metabolism , Chromatin Immunoprecipitation , Colonic Neoplasms/genetics , Colonic Neoplasms/metabolism , Cytoskeletal Proteins/genetics , Cytoskeletal Proteins/metabolism , Disease Progression , Down-Regulation , Epithelium/metabolism , Epithelium/pathology , Gene Expression Profiling , Homeodomain Proteins/genetics , Humans , Immunoblotting , Membrane Glycoproteins/genetics , Membrane Glycoproteins/metabolism , Membrane Proteins/genetics , Membrane Proteins/metabolism , Microscopy, Fluorescence , Middle Aged , Neoplasm Invasiveness/pathology , Nucleoside-Phosphate Kinase/genetics , Nucleoside-Phosphate Kinase/metabolism , Oligonucleotide Array Sequence Analysis , Promoter Regions, Genetic , Reverse Transcriptase Polymerase Chain Reaction , Snail Family Transcription Factors , Transcription Factors/genetics , Tumor Cells, Cultured , Zinc Finger E-box-Binding Homeobox 1ABSTRACT
We investigated the role of secretory leukocyte protease inhibitor (SLPI) in ischemia/reperfusion injury in cardiac transplantation. SLPI-/- mouse hearts and wild-type (WT) controls were transplanted immediately or after 10 h of cold ischemia (CI). Recombinant SLPI (rSLPI) was added to the preservation solution or given systemically. After evaluation of myocardial performance, grafts were investigated for histology, SLPI, TNF-alpha, TGF-beta, NF-kappaB and protease expression at indicated time points. Early myocardial contraction was profoundly impaired in SLPI-/- hearts exposed to CI and associated with high intra-graft protease expression. Systemic administration of rSLPI had no effect, however, when SLPI was added to the preservation solution, myocardial contraction was restored to normal. At 10 days, inflammation, myocyte vacuolization and necrosis were significantly more severe in SLPI-/- hearts. SLPI gene expression was detected in WT mice at 12 and 24 h and was significantly higher after CI. SLPI protein was observed at 24 h and 10 days. High intra-graft concentrations of SLPI after administration of rSLPI were inversely correlated with protease levels early and TGF-beta expression late after reperfusion. SLPI plays a crucial role in early myocardial performance and postischemic inflammation after cardiac transplantation. A dual inhibitory effect on protease and TGF-beta expression might be the underlying mechanism.
Subject(s)
Heart Transplantation/physiology , Secretory Leukocyte Peptidase Inhibitor/deficiency , Secretory Leukocyte Peptidase Inhibitor/therapeutic use , Animals , Blotting, Western , Enzyme-Linked Immunosorbent Assay , Heart Transplantation/methods , Heart Transplantation/pathology , Mice , Mice, Inbred C57BL , Mice, Knockout , Myocardial Contraction , Recombinant Proteins/therapeutic use , Reperfusion Injury , Reverse Transcriptase Polymerase Chain Reaction , Secretory Leukocyte Peptidase Inhibitor/genetics , Transforming Growth Factor beta/physiology , Transplantation, IsogeneicABSTRACT
Practice and accuracy of immunohistochemistry is known to vary highly. Reliability of HER-2 immunohistochemistry is critical because of its role in patient selection for therapeutical options in breast cancer. Therefore reliability of HER-2 immunohistochemistry in pathology laboratories in Austria was assessed. Ten tissue specimens of invasive ductal breast carcinomas and three cell line samples were tested. Presence/absence of gene amplification was determined by FISH to be used as a gold standard. Laboratories were asked to stain and assess slides using their routine immunohistochemical staining protocol. Overall the study consisted of 311 tests on tissue specimens and 142 on cell lines. In all cases manual scoring was performed. Participation was voluntary and was 94%. Overall sensitivity was 90.5% and specificity 99.2%. Overscoring including true false positive results were found in 6.7% and 6.3% in tissue specimens and cell lines, respectively. False negative determinations were obtained in 1.9% and 2.8% of tissue specimens and cell lines, respectively. HercepTest showed slightly higher reliability in comparison with individualized staining methods. By manual scoring inaccurate scoring affected 12.3% of test results and 62% of the laboratories. In conclusion participation rate and accuracy of HER-immunohistochemistry was high all over the country. Manually performed scoring demonstrated some limitations.
Subject(s)
Biomarkers, Tumor/metabolism , Breast Neoplasms/diagnosis , Breast Neoplasms/metabolism , Carcinoma, Ductal, Breast/diagnosis , Carcinoma, Ductal, Breast/metabolism , Immunohistochemistry/standards , Receptor, ErbB-2/metabolism , Austria , Breast Neoplasms/pathology , Carcinoma, Ductal, Breast/pathology , Cell Line, Tumor , False Negative Reactions , Female , Humans , Immunohistochemistry/methods , Quality Assurance, Health Care , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
BACKGROUND: Glioblastoma is a very aggressive brain tumour with poor prognosis despite radical surgery or radiotherapy. Signal transducers and activators of transcription (STAT) proteins are important elements in intracellular signalling and part of the JAK-STAT pathway. They are activated by growth factors and cytokines and translocate into the nucleus upon activation to exert their function as transcription factors. STAT-1 can be induced by interferons and has also been found to be important in sensitizing tumours to chemotherapeutic drugs. MATERIALS AND METHODS: Forty-six glioblastoma samples have been analysed for the expression of STAT-1 by immunohistochemistry. RESULTS: In our study performed by immunohistochemistry, 22 out of 46 glioblastomas (48%) were strongly positive for staining with a STAT-1 antibody, 9 (20%) showed an intermediate reactivity, 8 (17%) low immunoreactivity, and 7 (15%) were completely negative. In the tumour tissue, STAT-1 expression was mostly localized in the cytoplasm. This location of STAT-1 suggests the predominant presence of an inactive form of STAT-1. Tumour giant cells were frequently strongly stained. Part of the peritumoral brain tissue showed strongly positively reactive glial cells. Interestingly, within the infiltration area strong STAT-1 expression was found in reactive astrocytes, glia, and particularly in microglial components. CONCLUSION: The expression of STAT-1 in the majority of glioblastomas, together with its documented role in apoptosis and in the action of chemotherapeutic drugs on tumour cell lines point to a possible function of this protein in the response of glioblastomas to chemotherapy.
Subject(s)
Brain Neoplasms/metabolism , Glioblastoma/metabolism , STAT1 Transcription Factor/biosynthesis , Brain Neoplasms/pathology , Glioblastoma/pathology , Humans , ImmunohistochemistryABSTRACT
Heart rate (HR), systolic (SBP), and diastolic (DBP) blood pressure levels were compared in 60 young males during various resting and stressful conditions. Subjects included 29 who had and 31 who had not shown occasional casual SBP readings greater than or equal to 135 mm Hg under low stress conditions. These groups were further divided into those who showed above-average and below-average HR increases at onset of a stressful shock-avoidance task (high vs low HR reactors). High HR reactors, who were equally distributed in both casual SBP groups, also showed higher SBP than low reactors during avoidance, and higher HR and SBP during a pre-stress rest period and two other stresses, the cold pressor test and viewing an erotic film (p's less than 0.01). During less stressful conditions (relaxation in the laboratory, family doctor readings, and self-determinations at home), no HR, SBP, or DBP differences were seen between high and low HR reactors. Subjects with casual SBPs greater than or equal to 135 showed higher mean SBP than those with casual SBPs less than 135 under all resting and stressful conditions (p's less than 0.05) and generally higher DBP as well. Highest mean SBP levels during prestress rest and later stresses were shown by subjects with both casual SBPs greater than or equal to 135 and high HR reactivity to the avoidance task, and lowest by subjects with neither trait. Incidence of parental hypertension was greater among high than low HR reactors, and greatest among high reactors with casual SBPs greater than or equal to 135, suggesting that HR reactivity to stress may help predict future hypertension.
Subject(s)
Blood Pressure , Heart Rate , Myocardial Contraction , Stress, Psychological/physiopathology , Adolescent , Adult , Ambulatory Care , Avoidance Learning/physiology , Blood Pressure Determination/methods , Cold Temperature , Humans , Hypertension/etiology , Prospective Studies , Rest , Self Care , SexABSTRACT
A total of 228 men, aged 18 to 22 years (109 black and 119 white), underwent monitoring of heart rate (HR) and systolic (SBP) and diastolic blood pressure (DBP) responses during several stressor conditions and a 30-minute posttask rest period. Stressors included the cold pressor test and three reaction-time tasks: noncompetitive, competitive, and competitive plus money incentive. Substantial within-subject variations in blood pressure and heart rate were induced, varying from 119/70 to 148/94 mm Hg and from 63 to 91 beats/min on the average. Men (25 black and 39 white) with marginal SBP elevations during initial casual determinations had higher SBP under all conditions compared with men whose casual SBP levels were normal, and they also showed greater elevations over baseline levels in heart rate, SBP, and DBP during the stressors and the initial casual determination. Black and white subjects did not differ in their blood pressures at baseline or during the initial casual determinations, although blacks had slightly lower heart rates. Blacks did show greater SBP elevations over baseline levels than whites during the stressors, primarily because the blacks with marginally elevated SBP showed substantially greater stress-induced increases than whites with marginally elevated SBP. This enhanced pressor response to stress in blacks with marginal blood pressure elevations may be due to higher vascular resistance during enhanced sympathetic activity and could contribute to the higher incidence of hypertension among blacks.
Subject(s)
Blood Pressure , Hypertension/ethnology , Racial Groups , Stress, Physiological/physiopathology , Stress, Psychological/physiopathology , Adult , Black or African American , Heart Rate , Humans , Hypertension/physiopathology , Male , Reaction Time , United States , Vascular Resistance , White PeopleABSTRACT
Cancer-related fibrin deposition and fibrinolysis were investigated by two-dimensional gel electrophoresis of human solid tumor and effusion specimen in addition to plasma samples. Fibrinogen gamma-chain dimer indicating fibrin deposition and plasmin-generated fibrinogen beta-chain fragments were identified in various solid tumor types by amino acid sequencing, mass spectrometry analysis and Western blotting. In tumor-associated effusions, these techniques allowed to observe plasmin-generated fragments of fibrinogen alpha, beta and gamma-chains in addition to elevated levels of acute-phase proteins. Similar observations were made in case of inflammation-associated effusions. No fibrin degradation product was observed in plasma samples, however, high amounts of fibrinogen gamma-chain dimer crosslinked by transglutaminase were detected in plasma from tumor patients, but not in plasma from controls and patients suffering acute infections and/or inflammations. This finding demonstrated that high transglutaminase activity may be associated with cancer. The presented data indicate that the amount of crosslinked fibrinogen gamma-chain dimer in plasma may correlate with tumor-associated fibrin deposition. The tumor-biological relevance of this potential marker protein is discussed.
Subject(s)
Biomarkers, Tumor/metabolism , Cross-Linking Reagents/metabolism , Fibrinogen/metabolism , Neoplasms/diagnosis , Biomarkers, Tumor/blood , Case-Control Studies , Dimerization , Electrophoresis, Gel, Two-Dimensional , Fibrin/metabolism , Fibrinolysis , Hemostasis , Humans , Neoplasm Proteins/blood , Neoplasm Proteins/metabolism , Neoplasms/blood , Neoplasms/metabolism , Pleural Effusion, Malignant/metabolism , Proteome/analysis , Tissue DistributionABSTRACT
Growth factors are essential for cellular growth and differentiation in both normal and malignant human breast epithelial cells. In the present study we investigated the effect of epidermal growth factor (EGF), transforming growth factor alpha (TGFalpha) and phorbol myristate acetate (PMA) on chicken ovalbumin upstream promoter-transcription factor (COUP-TF) expression in human breast cancer cells. The orphan receptors COUP-TFI and COUP-TFII are members of the nuclear receptor superfamily. The high degree of evolutionary conservation of these proteins strongly argues for an important biological function. COUP-TF expression was highest in SK-BR3 cells (approximately 130 amol/ micro g total RNA), while the lowest COUP-TF expression was observed in MCF-7 cells (3.5 amol/ micro g total RNA). While treatment of EGF, TGFalpha and PMA induced expression of COUP-TFII, COUP-TFI did not respond to these agents. Oncostatin M (OSM) is known to exert an antiproliferative effect in breast cancer cells. Treatment of MCF-7 cells with OSM resulted in an approximately 90% reduction of COUP-TFII mRNA expression. In SK-BR3 cells, treatment with the MEK inhibitor UO126 resulted in a profound suppression of endogenous COUP-TFII expression. Furthermore, cotreatment with UO126 prevented induction of COUP-TFII expression by EGF in MCF-7 cells. In conclusion, our data provide evidence, for the first time, that mitogenic substances which activate the MAP kinase pathway, can induce COUP-TFII expression. Our results strongly suggest that an active MAP kinase pathway is essential for COUP-TFII expression in human breast cancer cells.
Subject(s)
Breast Neoplasms/metabolism , DNA-Binding Proteins/metabolism , MAP Kinase Signaling System , Receptors, Steroid , Transcription Factors/metabolism , Blotting, Western/methods , Butadienes/pharmacology , COUP Transcription Factor I , COUP Transcription Factor II , COUP Transcription Factors , DNA-Binding Proteins/analysis , DNA-Binding Proteins/genetics , Enzyme Activation , Enzyme Inhibitors/pharmacology , Epidermal Growth Factor/pharmacology , Female , Glyceraldehyde-3-Phosphate Dehydrogenases/analysis , Glyceraldehyde-3-Phosphate Dehydrogenases/genetics , Humans , Mitogen-Activated Protein Kinases/analysis , Mitogen-Activated Protein Kinases/antagonists & inhibitors , Nitriles/pharmacology , Oncostatin M , Peptides/pharmacology , Reverse Transcriptase Polymerase Chain Reaction , Tetradecanoylphorbol Acetate/pharmacology , Transcription Factors/analysis , Transcription Factors/genetics , Transforming Growth Factor alpha/pharmacology , Tumor Cells, CulturedABSTRACT
Sertoli-Leydig cell tumors (SLCT) are rare sex-cord stromal tumors of the ovary composed of undifferentiated gonadal stromal cells, Leydig cells (LC), and Sertoli cells (SC), with the latter forming structures resembling fetal testicular tubules. The histogenetic basis of morphological male differentiation patterns in females is controversial. Here, we report a SLCT with intermediate differentiation in a 23-year-old woman investigated by light microscopy, immunohistochemistry for intermediate filaments, and sex steroid hormone receptors (SSHR), as well as by polymerase chain reaction (PCR) for the presence of the sex-determining region Y gene (SRY). Our investigation shows that the SCs of SLCT express progesterone and androgen receptors as well as cytokeratins and vimentin. By PCR, SLCT-derived genomic DNA lacked the SRY gene, indicating that the SLCT results from a SRY gene-independent pathway of pseudomale gonadal differentiation. The expression of progesterone receptors (PRs) in the SCs of the SLCT is in contrast to their absence in testicular SCs, but in line with their presence in ovarian granulosa and surface epithelial cells. Thus, our results provide strong evidence for a close histogenetic relationship between the SLCT and the female gonocyte-supporting cell, the granulosa cell (GC).
Subject(s)
Nuclear Proteins , Ovarian Neoplasms/pathology , Sertoli-Leydig Cell Tumor/pathology , Transcription Factors , Adult , DNA Primers/chemistry , DNA-Binding Proteins/analysis , Female , Humans , Immunohistochemistry , Intermediate Filaments/immunology , Keratins/analysis , Ovarian Neoplasms/chemistry , Ovarian Neoplasms/genetics , Polymerase Chain Reaction , Receptors, Progesterone/blood , Sertoli-Leydig Cell Tumor/chemistry , Sertoli-Leydig Cell Tumor/genetics , Sex Determination Processes , Sex Differentiation/genetics , Sex-Determining Region Y Protein , Vimentin/analysisABSTRACT
Posttransplant lymphoproliferative disorders (PTLPDs) are predominantly B-cell lymphoproliferations, whereas a T-cell origin is rarely observed. In contrast to B-cell PTLPD, T-cell PTLPDs show an inconsistent association with Epstein-Barr virus (EBV). Until now, only 13 cases of EBV-associated T-cell PTLPDs have been reported. We describe a case of an EBV-associated T-cell PTLPD in a renal allograft recipient 2 years after transplantation. Histologic examination showed medium- to large-sized lymphoid cells with an angiocentric growth pattern and necrosis. The atypical cells showed a CD2+, CD3epsilon+, CD7+, CD43+, CD45R0+, CD56+, and CD4-, CD5-, CD8- betaF1- phenotype with expression of the latent membrane protein (LMP)-1 of EBV. In addition, EBV-specific RNAs (EBER 1/2) were identified by in situ hybridization. Molecular analysis of the T-cell receptor (TCR) gamma chain by polymerase chain reaction (PCR) showed a polyclonal pattern. The morphologic, immunohistochemical, and molecular findings were consistent with a diagnosis of an EBV-associated extranodal natural killer (NK)/T-cell non-Hodgkin lymphoma (NHL) of nasal type. To our knowledge, this is the first reported case of this rare entity in the posttransplant setting.
Subject(s)
Epstein-Barr Virus Infections/diagnosis , Hypopharynx , Kidney Transplantation , Killer Cells, Natural , Lymphoma, T-Cell/diagnosis , Opportunistic Infections/diagnosis , Pharyngeal Neoplasms/diagnosis , Aged , Antigens, CD/analysis , Antigens, CD/immunology , Epstein-Barr Virus Infections/classification , Epstein-Barr Virus Infections/virology , Herpesvirus 4, Human/genetics , Humans , Hypopharynx/pathology , Immunohistochemistry , Immunophenotyping , Lymphoma, T-Cell/classification , Lymphoma, T-Cell/virology , Male , Nasopharynx , Opportunistic Infections/classification , Opportunistic Infections/virology , Pharyngeal Neoplasms/classification , Pharyngeal Neoplasms/virology , RNA, Viral/analysisABSTRACT
We tested the 4-amino analogue of tetrahydrobiopterin (H(4)aminobiopterin), a novel pterin-based inhibitor of nitric oxide synthases, for its efficacy in a murine cardiac-transplant model employing an improved cuff technique. We treated groups of 5 animals each for the first 7 post-operative days with various doses of H(4)aminobiopterin, with Cyclosporin A (15 mg/kg/day), or no treatment. H(4)aminobiopterin (3 times 50 mg/kg/day) proved to be as efficient as high-dose Cyclosporin A (15 mg/kg/day) in prolonging allograft survival and in suppressing histologic changes caused by the immunoreaction. Surprisingly, the doses of H(4)aminobiopterin effective in prolonging allograft survival did not change the plasma nitrite plus nitrate, or the expression of inducible nitric oxide synthase, interferon-gamma, tumor necrosis factor-alpha, and B7-1 (CD80), indicating that H(4)aminobiopterin may act through a novel, yet undiscovered mechanism.
Subject(s)
Biopterins/pharmacology , Enzyme Inhibitors/pharmacology , Graft Survival/drug effects , Heart Transplantation , Nitric Oxide Synthase/antagonists & inhibitors , Animals , Biopterins/analogs & derivatives , Biopterins/pharmacokinetics , Cyclosporine/administration & dosage , Cyclosporine/pharmacokinetics , Dose-Response Relationship, Drug , Enzyme Inhibitors/pharmacokinetics , Half-Life , Immunosuppressive Agents/administration & dosage , Immunosuppressive Agents/pharmacokinetics , Interferon-gamma/analysis , Interferon-gamma/drug effects , Male , Mice , Mice, Inbred C3H , Mice, Inbred C57BL , Myocardial Revascularization/methods , Myocardium/pathology , Necrosis , Transplantation Tolerance/drug effectsABSTRACT
BACKGROUND & AIMS: DMT1 is a transmembrane protein which transports the divalent metal ions Fe2+, Zn2+, Cu2+ and Mn2+. Although DMT1 has been functionally linked to duodenal absorption and cellular utilisation of iron hardly anything is known about its distribution and potential role within the human glandular system. METHODS: Two polyclonal antibodies were raised to study the expression of DMT1 in tissues obtained from human corpus by the means of immunocytochemistry and Western blotting. RESULTS: All antibodies specifically detected a 60 kD protein band referring to human DMT1. Significant amounts of DMT1 expression were detected on the luminal site of organs, which are involved in excretion/re-absorption processes, such as salivary glands, pancreas, biliary tract and gallbladder. CONCLUSIONS: Our results suggest that DMT1 may be of pivotal importance for the regulation of metal ion homeostasis within organs involved in absorption and excretion of ions.
Subject(s)
Cation Transport Proteins/metabolism , Endocrine Glands/metabolism , Iron-Binding Proteins/metabolism , Animals , Antibody Specificity , Blotting, Western , Digestive System/metabolism , Exocrine Glands/metabolism , Humans , Immunohistochemistry , Kidney/metabolism , Rabbits/immunology , Tissue DistributionABSTRACT
To investigate the prognostic value of Her2/neu expression in differentiated thyroid carcinomas 103 patients were retrospectively investigated. All of them received surgical and an identical follow-up treatment. The patients with papillary and follicular thyroid cancer were further separated into two groups concerning their clinical development, including one group without distant metastasis (follow-up of minimum 8 years). The second group presented with distant metastases as a sign of an aggressive behaviour. Her2/neu was immunohistochemically detected on sections from formalin-fixed, paraffin-embedded tissues using c-erbB-2/Her-2/neu oncoprotein Ab-17 monoclonal antibody (mAb). In statistical analysis using the Mann-Whitney U-test and chi(2) test, Her2/neu protein overexpression was significantly correlated with prognosis. Both tumour entities without distant metastases showed significantly less cytoplasmic immunostaining than patients with development of metastases. Concerning the clinical outcome, Her2/neu overexpression may be regarded as a prognostic factor in differentiated thyroid carcinomas. Moreover, in addition to standard radio-iodine elimination therapy, application of Herceptin could lead to new successful therapeutic concepts for a number of patients with progressive thyroid cancer.
Subject(s)
Adenocarcinoma, Follicular/metabolism , Carcinoma, Papillary/metabolism , Receptor, ErbB-2/metabolism , Thyroid Neoplasms/metabolism , Adenocarcinoma, Follicular/secondary , Adenocarcinoma, Follicular/surgery , Carcinoma, Papillary/secondary , Carcinoma, Papillary/surgery , Cell Count , Female , Humans , Immunohistochemistry , In Situ Hybridization, Fluorescence , Male , Receptor, ErbB-2/genetics , Retrospective Studies , Single-Blind Method , Thyroid Neoplasms/pathology , Thyroid Neoplasms/surgeryABSTRACT
AIMS: Tetranectin (TN), a plasminogen kringle 4 binding protein, is thought to play a prominent role in the regulation of proteolytic processes via binding to plasminogen. The aim of this study was to evaluate the expression of TN in human breast cancer and adjacent normal breast tissue and to determine the impact of this expression on survival. METHODS: A retrospective analysis was performed on 189 patients with breast cancer, with a median follow up time of 10.6 years. The expression of TN was assessed in tumour tissue and adjacent normal breast tissue by immunohistochemistry, and the prognostic relevance of its expression in tumour cells was evaluated. RESULTS: TN was highly expressed in connective tissue fibres surrounding normal breast epithelium, but not in normal epithelial cells. High expression of TN in tumour cells was found in 131 (69%) of the tumour samples. By western blot analysis, no significant difference in the amount and molecular weight of TN was seen between tumour tissue and normal tissue. Strong TN immunoreactivity in tumour tissue was predictive of poor disease free and tumour specific overall survival. By multivariate analysis, high TN expression in cancer cells was an independent prognostic factor for disease free and tumour specific overall survival. CONCLUSIONS: Our results demonstrate differential TN expression in normal and malignant breast tissue and a prognostic impact of TN protein expression in breast carcinoma tissue. These data suggest a possible role of TN in invasiveness and the metastatic spread of human breast cancer.
Subject(s)
Breast Neoplasms/chemistry , Breast/chemistry , Carcinoma/chemistry , Lectins, C-Type/analysis , Adult , Aged , Aged, 80 and over , Breast Neoplasms/mortality , Carcinoma/mortality , Epidemiologic Methods , Female , Humans , Immunoblotting/methods , Immunohistochemistry/methods , Middle Aged , PrognosisABSTRACT
Alterations in blood pressure (BP) during two aversive behavioral tasks were studied in five chronically-prepared dogs. During a signalled-avoidance task, BP levels were not altered, although heart rate (HR) increased. While propranolol (0.3 mg/kg, IA) led to slight increases in resting pressure, and phenoxybenzamine (1.0 mg/kg) reductions, the tachycardia at avoidance onset was not affected. Exposure to an unsignalled-avoidance task led to elevated diastolic BP levels during a preavoidance period and to increases in systolic BP, HR and aortic dP/dt at the inception of the avoidance session. Again, neither drug affected the tachycardia during avoidance, but both agents precluded BP and aortic dP/dt increases. Patterns of intercorrelations among cardiovascular variables were similar for both tasks, and suggested that the basis of the BP maintenance shifted from vasomotor to cardiac control during the avoidance periods. The differential cardiovascular adjustments during these tasks could not be accounted for in terms of differences in response rate. Rather, the critical variable seemed to be the amount of feedback the animal received for responding.