72 research outputs found
Immunohistochemical analysis of estrogen receptors in breast carcinomas using monoclonal antibodies that recognize different domains of the receptor molecule
No full textEstrogen receptor (ER) analysis was performed in 46 primary breast carcinomas using four monoclonal antibodies (MABs) to ER (AER311, ER1D5, LH1, and LH2), each of which recognizes a distinct domain of the receptor protein. ER was expressed as the percentage of positively stained tumor cells. Statistical analysis was performed using the SPSS/PC+ program to set the cut off of positivity and the prognostic value of each MAB. A positivity >30% for each MAB possessed the best sensitivity/specificity ratio and was used as the cut-off value. Multivariate discriminant analysis showed that MABs AER311, ER1D5, and LH2 had significant prognostic value. Fourteen tumors showed positivity for these three MABs; 17 were positive for one or two of the three MABs, and 15 were negative for all three MABs. Survival analysis showed that patients with tumors negative for all three of these MABs had progression of the disease within 8 years from the diagnosis of the tumor, whereas all patients with tumors positive for all three MABs were alive 13 years after surgery. A significant correlation (P = 0.0006) between tumor grading and ER status was found; 71% of the tumors that were positive for all three MABs were grade 1, whereas tumors negative for all three MABs were mostly grades 2 and 3. No significant relationship was observed between ER status and tumor size. A significant correlation (P = 0.008) between lymph node status and ER was found; breast tumors positive for all three MABs were in the majority (92.9%) of cases pNO, whereas 67% of tumors negative for all three MABs were pN1. Results from the present study suggest that the use of a panel of MABs that target distinct epitopes within domains of the ER protein could offer a better approach for assessing the ER status in breast cancer patients, because it enables the recognition of breast tumors with intact or structurally defective ER proteins
CALDER: Cryogenic light detectors for background-free searches
No full textCALDER is a R&D project for the development of cryogenic light detectors with an active surface of 5x5cm2 and an energy resolution of 20 eV RMS for visible and UV photons. These devices can enhance the sensitivity of next generation large mass bolometric detectors for rare event searches, providing an active background rejection method based on particle discrimination. A CALDER detector is composed by a large area Si absorber substrate with superconducting kinetic inductance detectors (KIDs) deposited on it. The substrate converts the incoming light into athermal phonons, that are then sensed by the KIDs. KID technology combine fabrication simplicity with natural attitude to frequency-domain multiplexing, making it an ideal candidate for a large scale bolometric experiments. We will give an overview of the CALDER project and show the performances obtained with prototype detectors both in terms of energy resolution and efficiency
An inducible cell line (Natasha), from a neuroblastoma patient with circulating HSR-positive blasts, expressing neurohormones
No full textA cell line, established from a neuroblastoma patient, expresses NCAM and L1 cell adhesion molecules. Two chromosomal abnormalities were present in bone marrow (10%) and cell line (82%) metaphases: (i) a homogeneously staining region (HSR) at the distal part of chromosome 14, and (ii) an insertion of unidentified dark G-banding material in 1 p36. The identification in the patient of chr 14-HSR-positive tumour cells, before the in vitro adaptation, suggests a direct HSR formation without preceding double minutes (dms; or a very early in vivo dms----HSR transformation). N-myc was amplified in the HSR. Cells expressed proopiomelanocortin and corticotropin releasing factor mRNAs. Untreated cells were relatively differentiated; nevertheless they dramatically responded to retinoic acid, forming extensive neurites, growth-cones, cell-cell and cell-neurite junctions. Neurofilaments and synaptic figures containing many dense core granules were identified. This differentiation was irreversible. This cell line is therefore useful for the study of differentiation and in particular for the involvement of neurohormones in the differentiation process
Sampling of proximal and distal duodenal biopsies in the diagnosis and monitoring of celiac disease
No full textSince celiac disease-associated mucosal lesions are patchy, the diagnosis of the disease requires histological evaluation of multiple duodenal biopsies
Interleukin-21 sustains inflammatory signals that contribute to sporadic colon tumorigenesis
No full textThis is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.This work received support from the “Fondazione Umberto di Mario ONLUS”, Rome, and AIRC (MFAG-12108 to CS and IG-13049 to GM)
Aryl hydrocarbon receptor-driven signals inhibit collagen synthesis in the gut
No full textFibrostrictures (FS) are a major complication of Crohn's disease (CD). Pathogenesis of FS is not fully understood, but activation of fibroblasts and excessive collagen deposition are crucial in the development of FS. Here, we investigated the role of aryl hydrocarbon receptor (AhR) in intestinal fibrosis. AhR RNA and protein expression were evaluated in intestinal fibroblasts of CD patients and controls. CD fibroblasts were stimulated with TGF-β1 or TNF-α in the presence or absence of the AhR activator Ficz, an AhR antagonist CH223191, or a specific AhR-silencing RNA. In CD fibroblasts, TGF-β1 and TNF-α increased Col1A1, Col3A1 and α-SMA transcripts and collagen secretion and this effect was reduced by Ficz and upregulated by CH22319. TGF-β1 or TNF-α induced activation of p38 and ERK1/2 MAP kinases was decreased by Ficz and increased by CH223191. The inhibitory effect of Ficz on Map kinase activation and collagen induction was abolished by AhR silencing. To assess the role of AhR in vivo, mice with trinitrobenzene-sulfonic-acid induced colonic fibrosis were given Ficz or CH223191. Mice given either Ficz or CH223191 produced less or more collagen respectively as compared with control mice. Our results indicate that AhR is a negative regulator of profibrotic signals in the gut
Cadherin-11 is a Regulator of Intestinal Fibrosis
No full textBACKGROUND AND AIM: Although the mechanisms underlying the formation of intestinal fibrostrictures in Crohn's disease (CD) are not fully understood, activation of fibroblasts and excessive collagen deposition are supposed to contribute to the development of such complications. Here, we investigated the role of cadherin-11 (CDH-11), a fibroblast-derived protein that induces collagen production in various organs, in intestinal fibrosis. METHODS: CDH-11 expression was evaluated in inflammatory (I) and fibrostricturing (FS) CD mucosal samples, ulcerative colitis (UC) mucosal samples and ileal and colonic control samples by real-time PCR, Western blotting and immunohistochemistry. CDH-11 expression was evaluated in normal and CD intestinal fibroblasts stimulated with inflammatory/fibrogenic cytokines. FS CD fibroblasts were cultured either with a specific CDH-11 antisense oligonucleotide (AS) or activating CDH-11 fusion protein and activation of RhoA/ROCK and TGF-β pathways and collagen production were evaluated by Western blotting. Finally, we assessed the susceptibility of CDH-11-knockout (KO) mice to colitis-induced intestinal fibrosis. RESULTS: CDH-11 RNA and protein expression was increased in both CD and UC as compared to controls. In CD, the greater expression of CDH-11 was seen in FS samples. Stimulation of fibroblasts with TNF-α, interleukin (IL)-6, IFN-γ, IL-13 and IL-1β enhanced CDH-11 expression. Knockdown of CDH-11 in FS CD fibroblasts impaired RhoA/ROCK/TGF-β signalling and reduced collagen synthesis, while activation of CDH-11 increased collagen secretion. CDH-11 KO mice were largely protected from intestinal fibrosis. CONCLUSIONS: Data show that CDH-11 expression is up-regulated in IBD and suggest a role for this protein in the control of intestinal fibrosis
Protein unlocking procedures of formalin-fixed paraffin-embedded tissues: application to MALDI-TOF imaging MS investigations
No full textArchival formalin-fixed paraffin-embedded (FFPE) tissues are a powerful tool for examining the clinical course of diseases. These specimens represent an incredible mine of valuable clinical and biological information for proteomic investigation. MALDI-TOF imaging MS (MALDI-IMS) is a protein profiling technique which enables the direct sampling of histological section; however, the quality of molecular data are strongly influenced by the tissue preparation condition. In fact, in previous years most of the studies employing such a technological platform have been conducted using cryo-preserved tissues. We have developed an in vitro approach using "tissue surrogate" samples in order to explore different protein unlocking procedures which might enable a suitable recovery of polypeptides for MS analysis. The developed protocols have been compared both by MALDI-TOF MS and nLC-MS(E) analysis either on surrogate samples or on FFPE specimen from human breast cancer. The collected evidence has been applied for the preparation of FFPE tissue sections following MALDI-IMS analysis. Our results outline the possibility to obtain valuable peptide mass spectra profiles form FFPE preparations by applying a combined two steps procedure of heat induced antigen retrieval (HIAR) in presence of EDTA and on target trypsin hydrolysis. A multivariate statistical evaluation is presented and discussed according to molecular spatial distributions and tissue morphology
Characterisation of a human glioblastoma cell line (LI) expressing hypothalamic and pituitary hormones
No full textThe human glioblastoma cell line LI showed morphological features typical of its neuroectodermal origin. Cells were positive by immunofluorescence to GFAP, MHC class II, and L1 determinants. Cytogenetic analysis showed the presence of a modal chromosome number of 63, ranging from 58 to 69 chromosomes (DNA index was 1.6). Northern blot analysis demonstrated the presence of mRNA transcripts specific for transglutaminase C (type II or "tissue"), growth-hormone releasing-hormone (GHRH), insulin-like growth factor II (IGF-II), and proopiomelanocortin (POMC). The GHRH mRNA was present in two different sizes, one similar to the normal hypothalamic species of 0.75 kb, whilst the second species was a large transcript of approximately 10 kb size. Treatment with 5 microM retinoic acid or 5 mM alpha-difluoromethylornithine for 5 days sharply reduced the growth rate and also induced modulation of the ultrastructure and antigenic profile. This cell line may be useful to study glial differentiation and the relationship of GHRH, IGF-II and POMC expression with differentiation in neuroectodermal tumours
Cytoskeletal proteins and resident flora
No full textRecent observations demonstrate that enteropathogenetic and enterohaemorrhagic bacteria, as well as other non enteropathogenetic bacteria (Listeria, Coxiella Burnetii), may subvert the host cell cytoskeleton. Models from enteropathogenic bacteria demonstrate that cytoskeletal proteins are required for bacteria binding to the enterocytes and that they play a role in the immune response of the host to intestinal bacteria. The cytoskeletal protein family Tropomyosins is present in all eukaryotic cells, with multiple isoforms regulated by multiple genes. Of the different Tropomyosin isoforms, TM5 has been shown to be expressed in colonic and jejunal epithelial cells, while TM1 in colonic and jejunal smooth muscle. In vitro studies have shown the presence of serum and mucosal IgG against TM5 in almost two thirds of patients with ulcerative colitis, suggesting: a. a possible autoimmune response to Tropomyosin in these patients; b. the hypothesis that the development of pouchitis may be related to the expression of TM5 in the ileal pouch; c. the use of probiotics in the treatment of pouchitis. Overall, the new expression of cytoskeletal proteins on the cell surface appears to be possibly induced by several mechanisms, including intestinal bacteria and apoptosis. The expression of cytoskeletal proteins on the cell surface may induce tolerance or autoimmune response on target cells. Further investigations are, however needed on the possible role of cytoskeletal proteins in human diseases
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