scholarly journals Demonstration of alpha 1-antitrypsin by immunofluorescence on paraffin-embedded hepatic and pancreatic tissue.

1979 ◽  
Vol 27 (3) ◽  
pp. 794-796 ◽  
Author(s):  
M J McElrath ◽  
R M Galbraith ◽  
R C Allen
Keyword(s):  
Human Serum ◽  
Solid Phase ◽  
Islet Cells ◽  
Pancreatic Tissue ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Normal Human ◽  
Liver Biopsies ◽  
Alpha 1 Antitrypsin ◽  
Sepharose 4B

Studies were performed in an attempt to improve current immunohistological techniques for the demonstration of alpha 1-antitrypsin (A1AT) in formalin-fixed paraffin-embedded tissues. The unwanted fluorescence (UF) commonly occurring in such procedures was found to be effectively eliminated by immunoadsorption of A1AT antisera with human serum lacking A1AT (Pi-null phenotype) coupled in solid phase to glutaraldehyde-activated aminohexyl-Sepharose 4B. Specificity of the antisera for A1AT was established by subsequent solid phase immunoadsorption against normal human serum bound to AH-Sepharose 4B. Using these techniques, immunoreactive A1AT was demonstrated in the cytoplasm of hepatocytes in liver biopsies obtained from patients with Z and MZ serum phenotypes, and in the cytoplasm of normal pancreatic islet cells.

scholarly journals Immunohistochemical Characterization of Canine Transmissible Venereal Tumor

1996 ◽  
Vol 33 (3) ◽  
pp. 257-263 ◽  
Author(s):  
E. Mozos ◽  
A. Méndez ◽  
J. C. Gómez-Villamandos ◽  
J. Martín de las Mulas ◽  
J. Pérez
Keyword(s):  
Light Chain ◽  
B Lymphocyte ◽  
Protein S ◽  
Lambda Light Chain ◽  
Formalin Fixed Paraffin ◽  
S 100 ◽  
Formalin Fixed Paraffin Embedded ◽  
Progressive Growth ◽  
Staining Techniques ◽  
Alpha 1 Antitrypsin

The collective immunohistochemical expression of human lysozyme, human alpha-1-antitrypsin, human CD3 antigen, calf vimentin, human keratins, human lambda light chains, canine immunoglobulins IgG, IgM, and bovine protein S-100 has been analyzed on formalin-fixed, paraffin-embedded tissue sections of 25 spontaneous canine transmissible venereal tumors (CTVT) from both genital and extragenital locations using the avidin-biotin-peroxidase complex technique. Lysozyme immunoreactivity was detected in 10/25 CTVT, alpha-1-antitrypsin in 14/25 CTVT, and vimentin in 25/25 CTVT. All CTVT cells were negative to keratins 5 + 8 of the Moll catalogue (RCK-102), S-100 protein, lambda light-chain immunoglobulins, IgG, IgM, and CD3 antigen. The intratumoral T- and B-lymphocyte infiltrate was differentiated using CD3 antigen, lambda light-chain immunoglobulins, IgG, and IgM, and this technique could be useful to evaluate the regressive or progressive growth stage of venereal tumors. Our findings support the hypothesis of a histiocytic immunophenotype for CTVT, and these staining techniques could be used in the differential diagnosis with lymphomas.

scholarly journals Combinatorial transcription factor profiles predict mature and functional human islet α and β cells

2021 ◽  
Author(s):  
Shristi Shrestha ◽  
Diane C. Saunders ◽  
John T. Walker ◽  
Joan Camunas-Soler ◽  
Xiao-Qing Dai ◽  
...  
Keyword(s):  
Islet Cells ◽  
Hormone Secretion ◽  
Developmental State ◽  
Human Islets ◽  
Glucose Sensing ◽  
Pancreatic Tissue ◽  
Β Cells ◽  
Cellular Processes ◽  
Normal Human ◽  
Multiple Donors

ABSTRACTIslet-enriched transcription factors (TFs) exert broad control over cellular processes in pancreatic α and β cells and changes in their expression are associated with developmental state and diabetes. However, the implications of heterogeneity in TF expression across islet cell populations are not well understood. To define this TF heterogeneity and its consequences for cellular function, we profiled >40,000 cells from normal human islets by scRNA-seq and stratified α and β cells based on combinatorial TF expression. Subpopulations of islet cells co-expressing ARX/MAFB (α cells) and MAFA/MAFB (β cells) exhibited greater expression of key genes related to glucose sensing and hormone secretion relative to subpopulations expressing only one or neither TF. Moreover, all subpopulations were identified in native pancreatic tissue from multiple donors. By Patch-seq, MAFA/MAFB co-expressing β cells showed enhanced electrophysiological activity. Thus, these results indicate combinatorial TF expression in islet α and β cells predicts highly functional, mature subpopulations.

The Development of a Radioimmunoassay Procedure for the Estimation of Tamm—Horsfall Glycoprotein in Human Serum

1977 ◽  
Vol 52 (2) ◽  
pp. 183-191 ◽  
Author(s):  
P. J. G. Avis
Keyword(s):  
Human Serum ◽  
Serum Proteins ◽  
Solid Phase ◽  
Normal Serum ◽  
Small Sample ◽  
Agarose Beads ◽  
Assay Procedure ◽  
Series Of Experiments ◽  
Sepharose 4B ◽  
Cyanogen Bromide Activation

1. A quantitative radioimmunoassay was developed for the measurement of nanogram amounts of Tamm—Horsfall (TH) glycoprotein in the presence of serum proteins at concentrations above 30 mg/ml. 2. Specific anti-(TH-glycoprotein) antibodies were labelled with 125I and these were usable for a period of 8 weeks. 3. Agarose beads (Sepharose 4B), to which TH-glycoprotein had been coupled via cyanogen bromide activation of the Sepharose, were used as the solid phase in the assay. This proved stable upon storage at 4°C for periods in excess of 4 months. 4. The dissociation of the glycoprotein in the presence of serum proteins that was necessary for quantification was achieved by subjecting the sample to ultrasonication. 5. Assays conducted on a small sample of normal serum produced evidence that normal serum contained amounts (50–180 ng/ml) of a substance that reacted similarly to TH-glycoprotein in the assay procedure and in a series of experiments conducted to confirm the presence of this substance in human serum.

scholarly journals Chemiluminescence Quantitative Immunohistochemical Determination of MRP2 in Liver Biopsies

2005 ◽  
Vol 53 (12) ◽  
pp. 1451-1457 ◽  
Author(s):  
Massimo Guardigli ◽  
Marianna Marangi ◽  
Silvia Casanova ◽  
Walther F. Grigioni ◽  
Enrico Roda ◽  
...  
Keyword(s):  
Protein Expression ◽  
High Sensitivity ◽  
Clinical Samples ◽  
Biliary Cirrhosis ◽  
Common Procedure ◽  
Formalin Fixed Paraffin ◽  
Alternative Approach ◽  
Clinical Diagnoses ◽  
Formalin Fixed Paraffin Embedded ◽  
Liver Biopsies

Evaluation of protein expression in tissues and cells by electrophoretic and blotting techniques or by the quantification of the mRNA coding for the target protein is a common procedure in biochemistry research and clinical diagnoses. In this article, an alternative approach, based on an immunohistochemical procedure with chemiluminescent imaging detection, is described. The assay exploited the peculiar characteristics of the chemiluminescent detection of enzyme labels (high sensitivity and specificity, low background, easy quantification of the signal) for performing the direct, simple, and rapid quantitative evaluation of protein expression in tissues. When applied to the study of the levels of MRP2, a member of the human multidrug resistance-associated protein family, in samples obtained from formalin-fixed, paraffin-embedded liver biopsies, it allowed the reliable evaluation of the protein content of the tissue. Moreover, the analysis of clinical samples from patients with primary biliary cirrhosis under therapy with ursodeoxycholic acid gave results in line with those, previously reported in the literature, obtained with conventional protein expression analysis techniques.

scholarly journals Protein-embedding Technique: A Potential Approach to Standardization of Immunohistochemistry for Formalin-fixed, Paraffin-embedded Tissue Sections

2005 ◽  
Vol 53 (9) ◽  
pp. 1167-1170 ◽  
Author(s):  
Shan-Rong Shi ◽  
Cheng Liu ◽  
Jeanette Perez ◽  
Clive R. Taylor
Keyword(s):  
Solid Phase ◽  
Antigen Retrieval ◽  
Polymer Microsphere ◽  
Thin Sections ◽  
Tissue Sections ◽  
Embedding Technique ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Two Phases ◽  
Formalin Fixed

A serial study was performed to develop a protein-embedding technique for standardization of immunohistochemistry (IHC) on formalin-fixed, paraffin-embedded (FFPE) tissue sections. A protein carrier matrix must have two phases, a liquid phase to allow uniform mixing of a protein and a solid phase forming a ‘block’ that can be fixed and processed in the same manner as human tissue. This standardized protein block would serve as a source of thin sections for control of IHC and therefore must also withstand the boiling conditions of antigen retrieval (AR). After multiple experiments, a method was developed utilizing polymer microsphere (beads) as a support medium for protein. The method showed particular promise for quantitative IHC.

scholarly journals Investigation of Archived Formalin-Fixed Paraffin-Embedded Pancreatic Tissue with Whole-Genome Gene Expression Microarray

2011 ◽  
Vol 2011 ◽  
pp. 1-12 ◽  
Author(s):  
Nete V. Michelsen ◽  
Klaus Brusgaard ◽  
Qihua Tan ◽  
Mads Thomassen ◽  
Khalid Hussain ◽  
...  
Keyword(s):  
Gene Expression ◽  
Pancreatic Tissue ◽  
Ffpe Tissue ◽  
Congenital Hyperinsulinism ◽  
Whole Genome ◽  
Gene Expression Microarray ◽  
Expression Microarray ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

The use of formalin-fixed, paraffin-embedded (FFPE) tissue overcomes the most prominent issues related to research on relatively rare diseases: limited sample size, availability of control tissue, and time frame. The use of FFPE pancreatic tissue in GEM may be especially challenging due to its very high amounts of ribonucleases compared to other tissues/organs. In choosing pancreatic tissue, we therefore indirectly address the applicability of other FFPE tissues to gene expression microarray (GEM). GEM was performed on archived, routinely fixed, FFPE pancreatic tissue from patients with congenital hyperinsulinism (CHI), insulinoma, and deceased age-appropriate neonates, using whole-genome arrays. Although ribonuclease-rich, we obtained biologically relevant and disease-specific, significant genes; cancer-related genes; genes involved in (a) the regulation of insulin secretion and synthesis, (b) amino acid metabolism, and (c) calcium ion homeostasis. These results should encourage future research and GEM studies on FFPE tissue from the invaluable biobanks available at the departments of pathology worldwide.

scholarly journals Detection of N-Glycolyl GM3 Ganglioside in Neuroectodermal Tumors by Immunohistochemistry: An Attractive Vaccine Target for Aggressive Pediatric Cancer

2011 ◽  
Vol 2011 ◽  
pp. 1-6 ◽  
Author(s):  
Alejandra M. Scursoni ◽  
Laura Galluzzo ◽  
Sandra Camarero ◽  
Jessica Lopez ◽  
Fabiana Lubieniecki ◽  
...  
Keyword(s):  
Pediatric Cancer ◽  
Nonsmall Cell Lung Cancer ◽  
Potential Utility ◽  
Formalin Fixed Paraffin ◽  
Neuroectodermal Tumors ◽  
Formalin Fixed Paraffin Embedded ◽  
Normal Human ◽  
Secondary Antibodies ◽  
Membranous Staining ◽  
Formalin Fixed

The N-glycolylated ganglioside NeuGc-GM3 has been described in solid tumors such as breast carcinoma, nonsmall cell lung cancer, and melanoma, but is usually not detected in normal human cells. Our aim was to evaluate the presence of NeuGc-GM3 in pediatric neuroectodermal tumors by immunohistochemistry. Twenty-seven archival cases of neuroblastoma and Ewing sarcoma family of tumors (ESFT) were analyzed. Formalin-fixed, paraffin-embedded tumor samples were cut into 5 μm sections. The monoclonal antibody 14F7, a mouse IgG1 that specifically recognizes NeuGc-GM3, and a peroxidase-labeled polymer conjugated to secondary antibodies were used. Presence of NeuGc-GM3 was evident in 23 of 27 cases (85%), with an average of about 70% of positive tumors cells. Immunoreactivity was moderate to intense in most tumors, showing a diffuse cytoplasmic and membranous staining, although cases of ESFT demonstrated a fine granular cytoplasmic pattern. No significant differences were observed between neuroblastoma with and without NMYC oncogene amplification, suggesting that expression of NeuGc-GM3 is preserved in more aggressive cancers. Until now, the expression of N-glycolylated gangliosides in pediatric neuroectodermal tumors has not been investigated. The present study evidenced the expression of NeuGc-GM3 in a high proportion of neuroectodermal tumors, suggesting its potential utility as a specific target of immunotherapy.

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