scholarly journals Gastric MUC5AC and MUC6 are large oligomeric mucins that differ in size, glycosylation and tissue distribution

2002 ◽  
Vol 364 (1) ◽  
pp. 191-200 ◽  
Author(s):  
Henrik NORDMAN ◽  
Julia R. DAVIES ◽  
Gert LINDELL ◽  
Carme de BOLÓS ◽  
Francisco REAL ◽  
...  
Keyword(s):  
Polyclonal Antibodies ◽  
Gradient Centrifugation ◽  
Variable Number Tandem Repeat ◽  
Variable Number ◽  
Repeat Sequence ◽  
Surface Epithelium ◽  
Lewis Antigen ◽  
Gland Tissue ◽  
Before And After ◽  
Ion Exchange Hplc

Gastric MUC5AC and MUC6 mucins were studied using polyclonal antibodies. Immunohistochemistry showed MUC5AC to originate from the surface epithelium, whereas MUC6 was produced by the glands. Mucins from the surface epithelium or glands of corpus and antrum were purified using CsCl/4M guanidinium chloride density-gradient centrifugation. MUC5AC appeared as two distinct populations at 1.4 and 1.3g/ml, whereas MUC6, which was enriched in the gland tissue, appeared at 1.45g/ml. Reactivity with antibodies against the Leb structure (where Le represents the Lewis antigen) followed the MUC5AC distribution, whereas antibodies against the Ley structure and reactivity with the GlcNAc-selective Solanum tuberosum lectin coincided with MUC6, suggesting that the two mucins are glycosylated differently. Rate-zonal centrifugation of whole mucins and reduced subunits showed that both gastric MUC5AC and MUC6 are oligomeric glycoproteins composed of disulphide-bond linked subunits and that oligomeric MUC5AC was apparently smaller than MUC6. A heterogeneous population of ‘low-density’ MUC5AC mucins, which were smaller than the ‘high-density’ ones both before and after reduction, reacted with an antibody against a variable number tandem repeat sequence within MUC5AC, suggesting that they represent precursor forms of this mucin. Following ion-exchange HPLC, both MUC5AC and MUC6 appeared as several distinct populations, probably corresponding to ‘glycoforms’ of the mucins, the most highly charged of which were found in the gland tissue.

Microbiological Effects of a Routine Treatment for Decontaminating Hide-On Carcasses at a Large Beef Packing Plant

2015 ◽  
Vol 78 (2) ◽  
pp. 256-263 ◽  
Author(s):  
XIANQIN YANG ◽  
MADHU BADONI ◽  
FRANCES TRAN ◽  
COLIN O. GILL
Keyword(s):  
Escherichia Coli ◽  
Tandem Repeat ◽  
Variable Number Tandem Repeat ◽  
Variable Number ◽  
Routine Treatment ◽  
Multiple Locus ◽  
E Coli ◽  
Repeat Analysis ◽  
Before And After

To investigate the microbiological effects of a hide-on carcass decontaminating treatment recently implemented at a beef packing plant, carcasses undergoing routine processing at the plant were sampled during successive periods in January/February, April/May, and September/October. During each period, samples were collected from carcasses before and after the decontamination of hide-on carcasses, after skinning, before decontamination of the skinned carcasses, and at the end of the carcass dressing process. At each stage of processing during each period, samples were obtained by swabbing an area of 1,000 cm2 on each of 25 carcasses. Aerobes, coliforms, and Escherichia coli were enumerated. In most samples, coliforms were predominantly E. coli. In all three periods, the log mean numbers of aerobes and E. coli recovered from hides before decontamination were between 6.6 and 6.8 and between 5.3 and 5.9 log CFU/1,000 cm2, respectively. The log mean numbers of aerobes recovered from decontaminated hides were 6.6 log CFU/1,000 cm2 in January/February and April/May but 5.4 log CFU/1,000 cm2 in September/October. The log total numbers of E. coli recovered from decontaminated hides in January/February and April/May were 2.4 and 3.8 log CFU/25,000 cm2, respectively, but no E. coli was recovered from such carcasses in September/October. Log total numbers of aerobes and E. coli recovered from skinned or dressed carcasses were mostly >4 and between 1 and 2 log CFU/25,000 cm2, respectively. Typing of 480 E. coli isolates by multiple-locus variable-number tandem repeat analysis (MLVA) identified 218 MLVA types. Most isolates recovered from carcasses in different periods or at different stages of processing were of different MLVA types. However, small numbers of MLVA types were recovered in more than one period or from both hides before and after decontamination and skinned or dressed carcasses. The findings show that the hide-decontaminating treatment disrupted the usual transfer of E. coli from hides to meat surfaces during carcass skinning.

scholarly journals Mucus glycoproteins from pig gastric mucosa: identification of different mucin populations from the surface epithelium

1997 ◽  
Vol 326 (3) ◽  
pp. 903-910 ◽  
Author(s):  
Henrik NORDMAN ◽  
Julia R. DAVIES ◽  
Annkatrin HERRMANN ◽  
Niclas G. KARLSSON ◽  
Gunnar C. HANSSON ◽  
...  
Keyword(s):  
Gradient Centrifugation ◽  
Buoyant Density ◽  
High Density ◽  
Surface Epithelium ◽  
Low Density ◽  
Guanidinium Chloride ◽  
Gastric Mucus ◽  
High Iron ◽  
Cardiac Region ◽  
Ion Exchange Hplc

Pig gastric mucins were isolated from the surface epithelium of the cardiac region, corpus and antrum using density-gradient centrifugation after extraction in 6 M guanidinium chloride. In CsCl/0.5 M guanidinium chloride, mucins solubilized from the cardiac region appeared as a broad unimodal band at 1.52 g/ml whereas those from the corpus and antrum occurred as high- and low-density populations at 1.50 and 1.45 g/ml respectively. High-iron diamine reacted more strongly with the cardiac mucins and the high-density populations from corpus and antrum than with the two low-density ones. In keeping with this, approx. 60% of the oligosaccharides from the former mucins and 20% from the latter contained sulphate. All surface epithelial cells of the cardiac region stained with high-iron diamine, whereas in the corpus only the epithelium in the bottom of the pits reacted, suggesting that the high-density population from this region originates from these cells. Mucins from all regions were composed of subunits, each containing highly glycosylated domains. The mucins from the cardiac region were larger than those from the corpus and antrum, and reduced subunits as well as high-molecular-mass glycopeptides from the cardiac mucins were larger than the corresponding fragments from the other regions. Ion-exchange HPLC showed that reduced subunits from the cardiac mucins and the high-density populations from the corpus and antrum were more ‘acidic’ than reduced subunits from the two low-density ones. All mucins contained a ‘neutral’ fraction, in particular those from the antrum. Pig gastric mucus thus contains a number of distinctly different mucin populations varying in buoyant density, size, ‘acidity’, glycosylation, sulphation and tissue origin.

scholarly journals MUC5B is a major gel-forming, oligomeric mucin from human salivary gland, respiratory tract and endocervix: identification of glycoforms and C-terminal cleavage

1998 ◽  
Vol 334 (3) ◽  
pp. 685-693 ◽  
Author(s):  
Claes WICKSTRÖM ◽  
Julia R. DAVIES ◽  
Gitte V. ERIKSEN ◽  
Enno C. I. VEERMAN ◽  
Ingemar CARLSTEDT
Keyword(s):  
Respiratory Tract ◽  
Gradient Centrifugation ◽  
Guanidinium Chloride ◽  
Charge Densities ◽  
Terminal Domain ◽  
Disulphide Bonds ◽  
Whole Saliva ◽  
Submucosal Glands ◽  
Before And After ◽  
Ion Exchange Hplc

Mucins from human whole saliva, as well as from respiratory- and cervical-tract secretions, were subjected to density-gradient centrifugation in CsCl/0.5 M guanidinium chloride. A polydisperse population of MUC5B mucins was demonstrated in all samples using anti-peptide antisera (LUM5B-2, LUM5B-3 and LUM5B-4) raised against sequences within the MUC5B mucin. The sequences recognized by the LUM5B-2 and LUM5B-3 antisera are located within the domains flanking the highly glycosylated regions of MUC5B, and reduction increased the reactivity with these antibodies, suggesting that the epitopes are partially shielded and that these regions are folded and stabilized by disulphide bonds. Rate-zonal centrifugation before and after reduction showed MUC5B to be a large oligomeric mucin composed of disulphide-linked subunits. In saliva and respiratory-tract secretions, populations of MUC5B mucins with different charge densities were identified by ion-exchange HPLC, suggesting the presence of MUC5B ‘glycoforms ’. In trachea, the F2 monoclonal antibody against the sulpho-Lewis C structure reacted preferentially with the later-to-be-eluted populations. An antibody (LUM5B-4) recognizing a sequence in the C-terminal domain of MUC5B identified, after reduction, the mucin subunits as well as smaller fragments, suggesting that some of the MUC5B mucins are cleaved within the C-terminal domain. Immunohistochemistry revealed that MUC5B is produced by cells dispersed throughout the human submandibular and sublingual glands, in the airway submucosal glands as well as the goblet cells, and in the epithelium and glands of the endocervix. The F2 antibody stained a subpopulation of the MUC5B-producing cells in the airway submucosal glands, suggesting that different cells may produce different glycoforms of MUC5B in this tissue.

scholarly journals Different mucins are produced by the surface epithelium and the submucosa in human trachea: identification of MUC5AC as a major mucin from the goblet cells

1996 ◽  
Vol 318 (1) ◽  
pp. 319-324 ◽  
Author(s):  
Hans W HOVENBERG ◽  
Julia R DAVIES ◽  
Ingemar CARLSTEDT
Keyword(s):  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Goblet Cells ◽  
Major Product ◽  
Surface Epithelium ◽  
Epithelial Surface ◽  
Predominant Species ◽  
Submucosal Glands ◽  
Ion Exchange Hplc ◽  
Human Trachea

Mucins were extracted from the epithelial surface and the submucosal tissue of human trachea in order to enrich glycoproteins from the goblet cells and the submucosal glands respectively. The macromolecules were purified using density-gradient centrifugation, and the presence of the MUC5AC mucin was investigated using an antiserum raised against a synthetic peptide based on the sequence of the MUC5AC apoprotein. Mucins from the surface epithelium showed a higher reactivity with the antiserum relative to carbohydrate than those from the submucosa, and ion-exchange HPLC of reduced subunits revealed the presence of two distinct mucin populations in the samples. The predominant species from the surface epithelium was more acidic than the major population from the submucosa and showed a strong reactivity with the anti-MUC5AC antiserum. In contrast, the major portion of the submucosal mucins were less acidic and showed no MUC5AC reactivity, although a more acidic population did react with the antibody. Rate-zonal centrifugation showed that the MUC5AC mucin from the surface epithelium is smaller than the major submucosal mucin, and that both are composed of subunits. Immunolocalization confirmed that the MUC5AC mucin from human trachea originates from the goblet cells and that this glycoprotein is not a major product of the submucosal glands.

Genetic Characterization of Listeria monocytogenes Isolates from Food Processing Facilities before and after Postcook Chiller Heat Treatment

2013 ◽  
Vol 76 (8) ◽  
pp. 1466-1470
Author(s):  
SOFRONI EGLEZOS ◽  
GARY A. DYKES ◽  
BIXING HUANG ◽  
MARK S. TURNER ◽  
RICHARD SEALE
Keyword(s):  
Heat Treatment ◽  
Listeria Monocytogenes ◽  
Variable Number Tandem Repeat ◽  
Variable Number ◽  
Heat Shock Gene ◽  
Class Iii ◽  
Before And After ◽  
Shock Gene ◽  
Mlva Typing

Possible selection for and establishment of stress-resistant Listeria monocytogenes variants as a consequence of heating interventions is of concern to the food industry. Lineage analysis and multilocus variable number tandem repeat analysis (MLVA) was performed on 20 L. monocytogenes isolates, of which 15 were obtained before and 5 were obtained after heat treatment of a postcook meat chiller. The ctsR gene (a class III heat shock gene regulator) from 14 isolates was amplified and sequenced because previous work has indicated that spontaneous mutations can occur in this gene during heat treatment. Heat treatment of the meat chiller did not significantly change the relative abundance of the various L. monocytogenes lineages; lineage II strains (less-heat-resistant isolates) dominated both before and after heat treatment. MLVA typing confirmed that some isolates of L. monocytogenes occur both before and after heat treatment of the chiller. No isolate of L. monocytogenes indicated any likely functionally significant mutations in ctsR. This study indicates the absence of any obvious difference in the profiles of L. monocytogenes strains obtained before and after heat treatment of a meat chiller, based on the characteristics examined. Although this finding supports the effectiveness of heat treatment, the limited number of strains used and characteristics examined mean that further study on a larger scale is required before firm conclusions can be drawn.

scholarly journals Diversity in a Variable-Number Tandem Repeat fromYersinia pestis

2000 ◽  
Vol 38 (4) ◽  
pp. 1516-1519 ◽  
Author(s):  
D. M. Adair ◽  
P. L. Worsham ◽  
K. K. Hill ◽  
A. M. Klevytska ◽  
P. J. Jackson ◽  
...  
Keyword(s):  
Tandem Repeat ◽  
Variable Number Tandem Repeat ◽  
Haplotype Diversity ◽  
Laboratory Culture ◽  
Culture Conditions ◽  
Variable Number ◽  
Repeat Sequence ◽  
Open Reading Frames ◽  
Tetranucleotide Repeat ◽  
Allele Type

We have identified a tetranucleotide repeat sequence, (CAAA)N, in the genome of Yersinia pestis, the causative agent of plague. This variable-number tandem repeat (VNTR) region has nine alleles and great diversity (calculated as 1 minus the sum of the squared allele frequencies) (diversity value, 0.82) within a set of 35 diverse Y. pestis strains. In contrast, the nucleotide sequence of the lcrV (low-calcium-response) gene differed only slightly among these strains, having a haplotype diversity value of 0.17. Replicated cultures, phenotypic variants of particular strains, and extensively cultured replicates within strains did not differ in VNTR allele type. Thus, while a high mutation rate must contribute to the great diversity of this locus, alleles appear stable under routine laboratory culture conditions. The classic three plague biovars did not have single identifying alleles, although there were allelic biases within biovar categories. The antiqua biovar was the most diverse, with four alleles observed in 5 strains, while the orientalis and mediaevalis biovars exhibited five alleles in 21 strains and three alleles in 8 strains, respectively. The CAAA VNTR is located immediately adjacent to the transcriptional promoters for flanking open reading frames and may affect their activity. This VNTR marker may provide a high-resolution tool for epidemiological analyses of plague.

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