Human intestinal goblet cell mucin

1976 ◽  
Vol 54 (8) ◽  
pp. 707-716 ◽  
Author(s):  
I. Jabbal ◽  
D. I. C. Kells ◽  
G. Forstner ◽  
J. Forstner
Keyword(s):  
Goblet Cell ◽  
High Speed ◽  
Proteolytic Enzymes ◽  
Periodic Acid ◽  
Periodic Acid Schiff ◽  
Strong Concentration ◽  
Acid Ratio ◽  
Schiff Reagent ◽  
Major Chemical ◽  
Sepharose 4B

Goblet cell mucin (GCM) has been purified for the first time from mucosal scrapings of human small intestine. Proteolytic enzymes and organic solvents were avoided during the isolation procedure. The mucin was purified by Sepharose 4B and 2B column chromatography of high-speed supernatant fractions. The most purified fraction was compared with rat intestinal GCM. The two were similar with respect to chemical composition, antigenic features, and polyacrylamide disc gel electrophoresis. The major chemical differences included a higher hexosamine–fucose and hexosamine – sialic acid ratio in human mucin. The two mucins showed strong concentration dependence in sedimentation velocity studies. Human mucin at a concentration of 0.2 to 1.5 mg protein per millilitre gave multiple associated peaks with variable s0 values (10.8–36.6). Rat mucin, in contrast, gave a constant (although polydisperse) pattern with s0 = 15.15. To explore these differences both mucins were stained with periodic acid – Schiff reagent and subjected to band ultracentrifugation at concentrations of 0.6–1.9 μg protein per millilitre. At this low concentration, rat mucin did not change in its sedimentation characteristics. In contrast, human GCM produced a single peak with s0 = 37.9. Thus dilution abolished polydispersity in the human but not the rat mucin, suggesting that intermolecular bonding forces in the human mucin are weaker.

Effects of matrix metalloproteinase inhibitor on LPS-induced goblet cell metaplasia

2004 ◽  
Vol 287 (1) ◽  
pp. L127-L133 ◽  
Author(s):  
Je Hyeong Kim ◽  
Sung Yong Lee ◽  
Sang Myeon Bak ◽  
In Bum Suh ◽  
Sang Yeub Lee ◽  
...  
Keyword(s):  
Goblet Cell ◽  
Bacterial Infections ◽  
Inflammatory Responses ◽  
Periodic Acid ◽  
Growth Factor Receptor ◽  
Sprague Dawley ◽  
Neutrophilic Infiltration ◽  
Periodic Acid Schiff ◽  
Mucus Hypersecretion ◽  
Egfr Expression

Bacterial infections of the lung are known to induce inflammatory responses, which lead to mucus hypersecretion. Moreover, mucin synthesis in the airways has been reported to be regulated by neutrophilic inflammation-induced epidermal growth factor receptor (EGFR) expression and its activation. Furthermore, matrix metalloproteinases (MMPs), especially MMP-9, have been reported to promote the transmigration of activated neutrophils. In this study, we investigated the associations between lipopolysaccharide (LPS)-induced goblet cell (GC) metaplasia and EGFR expression and the effects of MMP inhibitor (MMPI). Various concentrations of LPS were instilled into the tracheas of pathogen-free Sprague-Dawley rats, and airways were examined at different times after LPS instillation. To examine the role of MMP-9, we treated rats 3 days before LPS instillation and daily thereafter with MMPI. Neutrophilic infiltration, Alcian blue/periodic acid-Schiff (AB/PAS) staining, and immunohistochemical staining for MUC5AC, EGFR, and MMP-9 were performed. The instillation of LPS increased AB/PAS and MUC5AC staining in time- and dose-dependent manners, and treatment with MMPI significantly prevented GC metaplasia. The instillation of LPS into the trachea also induced neutrophilic infiltration and EGFR and MMP-9 expression in the airway epithelium, and MMPI was found to significantly prevent neutrophil recruitment, GC metaplasia, and EGFR and MMP-9 expression. This study demonstrates that the MMP-9 and EGFR cascades are associated with LPS-induced mucus hypersecretion.

α‎1-Antitrypsin deficiency and the serpinopathies

2020 ◽  
pp. 2235-2242
Author(s):  
David A. Lomas
Keyword(s):  
Conformational Transition ◽  
Proteolytic Enzymes ◽  
Periodic Acid ◽  
Cor Pulmonale ◽  
European Medicines Agency ◽  
Periodic Acid Schiff ◽  
Virtual Absence ◽  
Antitrypsin Deficiency ◽  
Excessive Alcohol ◽  
A Minor

α‎1-Antitrypsin is an acute phase glycoprotein synthesized by the liver that functions as an inhibitor of a range of proteolytic enzymes, most importantly neutrophil elastase in the lung. Ninety-five per cent of severe plasma deficiency of α‎1-antitrypsin results from homozygosity for the Z allele (Glu342Lys), which causes the protein to undergo a conformational transition and form ordered polymers that are retained within hepatocytes as periodic acid–Schiff-positive, diastase-resistant inclusions. Clinical features— all adults homozygous for the Z allele of α‎1-antitrypsin have a minor degree of portal fibrosis that is often subclinical, but up to 50% have clinically evident cirrhosis and occasionally hepatocellular carcinoma. They also develop panlobular emphysema that typically affects the lung bases and is greatly exacerbated by smoking. Cor pulmonale and polycythaemia are late features. Diagnosis and management—severe genetic deficiency of α‎1-antitrypsin is readily diagnosed by low plasma levels and the virtual absence of the α‎1-band on protein electrophoresis. Patients should abstain from smoking and avoid agents that cause hepatic injury, such as excessive alcohol and obesity. Emphysema is treated along conventional lines. α‎1-Antitrypsin replacement therapy is widely used in North America to slow the progression of the lung disease and has recently been licensed by the European Medicines Agency, but its clinical efficacy remains contentious and it has no effect on liver disease. Clinical trials are underway to ‘knock down’ the expression of mutant Z α‎1-antitrypsin within hepatocytes to try to prevent cirrhosis. Other serpinopathies—the polymerization that underlies α‎1-antitrypsin deficiency is found in other members of the serine protease inhibitor (or serpin) superfamily to cause diseases as diverse as thrombosis (antithrombin), angio-oedema (C1 inhibitor), and dementia (neuroserpin).

scholarly journals The Structural Characteristics of Chicken Fibrinogen

1977 ◽  
Author(s):  
J. Pindyck ◽  
M. W. Mosesson ◽  
D. Bannerjee ◽  
D. Galanakis
Keyword(s):  
Gel Electrophoresis ◽  
Structural Characteristics ◽  
Periodic Acid ◽  
Factor Xiiia ◽  
Periodic Acid Schiff ◽  
Polymer Formation ◽  
Sensitive Site ◽  
Schiff Reagent ◽  
The One ◽  
Fibrinogen Molecule

The structure and subunit composition of chicken fibrinogen(ϕ) have been investigated. Dodecyl sulfate gel electrophoresis of unreduced specimens revealed a single ϕ band with a molecular weight of approximately 320,000. ϕ and fibrin specimens were also electrophoresed after reduction with dithiothreitol, and after crosslinking of unreduced specimens in the presence of Factor Xllla. Chromatographically separated S-sulfo chains were also studied after reptilase or thrombin treatment,and certain samples were stained with periodic acid Schiff reagent(PAS). Chicken Aα chains weresmaller than human Aα chains (54,500 vs.70,900, respectively) but, like mammalian Aα chains, they possessed a reptilase and thrombin sensitive site, were PAS negative,and undergo Factor XIIIa catalyzed α-polymer formation. The sizes of chicken Bβ and γ chains were nearly thesame as their mammalian counterparts, (i. e. 60,000 and 49,000 respectively) ; both types of chains were PAS positive. Chicken Bβ chains possessed a slowly reactive thrombin sensitive site apparently corresponding to the one in human ϕ; the chicken β chains, like mammalian β chains, did not undergo Factor XIIIa catalyzed cross-linking. Like mammalian γ chains, chicken γ chains could undergo Factor XIIIa catalyzed γ-γ dimerization and did not possess thrombin or reptilase sensitive sites. These findings indicate that the chicken fibrinogen molecule is composed of three pairs of disulfide-bridged chains corresponding in most respects to mammalian fibrinogen chains.

scholarly journals IMMUNOLOGIC STUDIES OF HEART TISSUE

1961 ◽  
Vol 113 (1) ◽  
pp. 1-16 ◽  
Author(s):  
Melvin H. Kaplan ◽  
Frederick D. Dallenbach
Keyword(s):  
Heart Disease ◽  
Connective Tissue ◽  
Gamma Globulin ◽  
Fluorescent Antibody ◽  
Periodic Acid ◽  
Heart Tissue ◽  
Periodic Acid Schiff ◽  
Schiff Reagent ◽  
Points Of View ◽  
Rheumatic Heart

Using fluorescent antibody methods, deposits of bound gamma globulin, as determined in unfixed washed sections of auricular appendages from rheumatic hearts, were noted in a significant number (18 per cent) of 100 specimens studied. Such deposits were observed in myofibers, sarcolemma, interstitial connective tissue, and vessel walls. Albumin and fibrin were generally found absent from these sites. Control hearts from normal and pathologic material, including postmortem and biopsied specimens, in general, did not reveal such deposits. These various tissue sites which contained bound gamma globulin frequently exhibited evidence of alteration as indicated both by enhanced affinity for eosin and by strongly positive reaction with the periodic acid-Schiff reagent, and appeared comparable in some cases to "fibrinoid." Bound gamma globulin was not observed in cellular or stromal components of Aschoff lesions, nor was the occurrence of Aschoff lesions correlated with presence of bound gamma globulin. It is suggested that deposition of gamma globulin and the eosinophilic alteration associated with such deposition are related to certain of the pathologic changes of rheumatic heart disease. The nature of such deposits of gamma globulin was considered from immune and non-immune points of view.

The Properties of Soluble Proteins Synthesized by Nephrotic Rat Kidney Microsomes

1972 ◽  
Vol 50 (3) ◽  
pp. 292-298
Author(s):  
V. N. Katiyar ◽  
B. L. Dinh
Keyword(s):  
Gel Electrophoresis ◽  
Rat Kidney ◽  
Periodic Acid ◽  
Rabbit Antiserum ◽  
Periodic Acid Schiff ◽  
Rat Serum ◽  
Tissue Protein ◽  
Schiff Reagent ◽  
Normal Rat ◽  
Kidney Microsomes

The incorporation of 14C-leucine into the microsomal proteins of nephrotic rat kidney was much higher than that in the microsomal proteins of the normal rat kidney. When the newly synthesized microsomal proteins were analyzed by acrylamide gel electrophoresis, it was found that the higher incorporation of 14C-leucine in nephrotic rat kidney was mostly due to an increase in the biosynthesis of a tissue protein which migrated in the electrophoretic zone between serum albumin and transferrin. This protein did not react with rabbit antiserum to normal rat serum and was stained with periodic acid – Schiff reagent. It was believed to be excreted in the urine of nephrotic rats in great quantity and was easily identified as a distinct band in electrophoregrams of urine from these rats.

Isolation and partial characterization of a soluble mucin in human pancreatic juice

1991 ◽  
Vol 69 (8) ◽  
pp. 566-571
Author(s):  
S. P. Lee ◽  
Y. S. Choong ◽  
H. Z. Park
Keyword(s):  
Molecular Weight ◽  
Pancreatic Juice ◽  
Periodic Acid ◽  
Average Molecular Weight ◽  
Small Molecular Weight ◽  
Periodic Acid Schiff ◽  
Cellulose Acetate Electrophoresis ◽  
Cscl Density Gradient ◽  
Oligosaccharide Moiety ◽  
Schiff Reagent

Morphological and histochemical abnormalities in pancreatic mucin occur in many pancreatic disorders. However, the composition of pancreatic mucin is poorly understood. Purified mucin was isolated from pure pancreatic juice by sequential chromatography on Sepharose CL-2B and CL-4B followed by CsCl density gradient ultracentrifugation. The mucin preparation consists of 24% protein and 73% carbohydrate. Reduction of the macromolecule (> 2 × 106) by mercaptoethanol resulted in the formation of subunits of molecular weight 500 000 and released several small molecular weight proteins, including a glycoprotein of an average molecular weight of 116 000. Cellulose acetate electrophoresis separated the mucin into three species of different staining properties for periodic acid-Schiff reagent and Alcian blue, suggesting the presence of microheterogeneity with respect to sulphation and sialation. Threonine, serine, and proline composed 48% of the total amino acids, while the oligosaccharide moiety contained N-acetylglucosamine, N-acetyl-galactosamine, fucose, galactose, sialic acid, and sulphate. We also detected the presence of C16:0 and C18:0 fatty acids which were probably noncovalently bound to the pancreatic mucin.Key words: pancreas, mucin, glycoprotein, cystic fibrosis.

scholarly journals COMPARISON OF IMMUNOHISTOCHEMICAL MARKER WITH SPECIAL STAINS IN THE PROGNOSIS OF THE DIFFERENT GRADES OF ORAL SQUAMOUS CELL CARCINOMA

2019 ◽  
Vol 8 (4) ◽  
Author(s):  
Dr. Parthiban Nallaiyan ◽  
Dr. Bharathi Ramakrishnan ◽  
Dr. Gnanadeepam Santigo ◽  
Dr. Dhanalakshmi Jeyasivanesan
Keyword(s):  
Squamous Cell Carcinoma ◽  
Oral Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Squamous Cell ◽  
Periodic Acid ◽  
Collagen Fibre ◽  
Ki 67 ◽  
Periodic Acid Schiff ◽  
Schiff Reagent ◽  
Neutral Mucin

Objectives: The aim of the study is to compare the efficiency of Ki-67 with special stains in the stroma of different grades of Oral Squamous Cell Carcinoma (OSCC) and to evaluate the influence of these changes in predicting the prognosis of these tumors. Materials and Methods: A total of 30 cases of different grades of Oral Squamous Cell Carcinoma and 6 cases of control were sections and stained with Picrosirus red (PR), combined Alcian blue-periodic acid Schiff reagent (AB-PAS) and Immunohistochemical (IHC) marker Ki-67. Results: Collagen fibre nature using PR stain and proliferative activity of malignant epithelial cells using IHC marker Ki-67 was found to be statistically significant. Mucin presence using AB-PAS was statistically insignificant. Conclusion: Prognosis in different grades of oral squamous cell carcinoma can be accessed by change in collagen fibres birefringence, as the tumour progresses there is change from mature to immature collagen. Ki-67 is a good proliferative marker and shows that there is positive correlation with histological grading of oral squamous cell carcinoma. Presence of acidic or neutral mucin in OSCC needed to be further studied. For assessing the prognosis in different grades of OSCC, special stains can also be used. Keywords: Collagen, Mucin, Squamous cell carcinoma, Ki-67.

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