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Differentiation of mucinous from non-mucinous pancreatic cyst fluid using dual-stained, 1 dimensional polyacrylamide gel electrophoresis.

Streitz JM, Madden MT, Salo W, Bernadino KP, Deutsch JL, Deutsch JC - Clin Proteomics (2014)

Bottom Line: The gel was first stained with periodic acid Schiff (PAS), a mucin histochemical stain followed by a secondary protein staining with Simply Blue Safestain (Invitrogen).Visual scoring (based on the presence of mucins) gave a sensitivity of 95%, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 88% for prediction of mucinous histology.One dimensional SDS polyacrylamide gel electrophoresis of pancreatic cyst fluid, followed by mucin (PAS) and protein (Simply Blue Safestain) staining, provides a means of concentrating and visualizing mucins, which allows the accurate differentiation of mucinous from non-mucinous histology in pancreatic cysts.

View Article: PubMed Central - PubMed

Affiliation: Departments of Surgery, Essentia Health System/Duluth Clinic, University of Minnesota Medical School, Duluth, MN USA.

ABSTRACT

Background: Pancreatic cysts are being increasingly identified in patients. Mucinous cysts have malignant potential whereas non-mucinous cysts do not. Distinguishing potentially malignant cysts from harmless ones by the characterization of cyst fluid contents remains a difficult problem. This study was undertaken to determine whether cyst fluid mucin glycoprotein analysis could differentiate mucinous from non-mucinous pancreatic cysts.

Methods: Cyst fluid from 28 patients who underwent resection of a pancreatic cyst was used for the study. In each case the type of cyst was histologically identified. One dimensional SDS polyacrylamide gel electrophoresis (1D-SDS PAGE) was performed on cyst fluid samples. For the detection of the separated proteins, we employed a novel dual staining technique. The gel was first stained with periodic acid Schiff (PAS), a mucin histochemical stain followed by a secondary protein staining with Simply Blue Safestain (Invitrogen).

Results: Visual scoring (based on the presence of mucins) gave a sensitivity of 95%, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 88% for prediction of mucinous histology.

Conclusions: One dimensional SDS polyacrylamide gel electrophoresis of pancreatic cyst fluid, followed by mucin (PAS) and protein (Simply Blue Safestain) staining, provides a means of concentrating and visualizing mucins, which allows the accurate differentiation of mucinous from non-mucinous histology in pancreatic cysts.

No MeSH data available.


Related in: MedlinePlus

1-D SDS PAGE; primary PAS glycoprotein staining of representative pancreatic cyst fluid samples. (Direction of travel on the gel is downward).
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Fig1: 1-D SDS PAGE; primary PAS glycoprotein staining of representative pancreatic cyst fluid samples. (Direction of travel on the gel is downward).

Mentions: Primary staining was done with periodic acid-Schiff (PAS). Following electrophoresis the gel was immersed in 12.5% trichloracetic acid for 20 minutes, then 1% periodic acid for 25 minutes in the dark. It was then immersed in Schiff’s fuchsin-sulfite reagent (Sigma-Aldrich, St. Louis MO) for 15 minutes in the dark, washed in 0.5% metabisulfate twice and then washed in water until the gel destained. The gel was then photographed. As seen in Figure 1, results showed red staining of the glycoproteins.


Differentiation of mucinous from non-mucinous pancreatic cyst fluid using dual-stained, 1 dimensional polyacrylamide gel electrophoresis.

Streitz JM, Madden MT, Salo W, Bernadino KP, Deutsch JL, Deutsch JC - Clin Proteomics (2014)

1-D SDS PAGE; primary PAS glycoprotein staining of representative pancreatic cyst fluid samples. (Direction of travel on the gel is downward).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4363379&req=5

Fig1: 1-D SDS PAGE; primary PAS glycoprotein staining of representative pancreatic cyst fluid samples. (Direction of travel on the gel is downward).
Mentions: Primary staining was done with periodic acid-Schiff (PAS). Following electrophoresis the gel was immersed in 12.5% trichloracetic acid for 20 minutes, then 1% periodic acid for 25 minutes in the dark. It was then immersed in Schiff’s fuchsin-sulfite reagent (Sigma-Aldrich, St. Louis MO) for 15 minutes in the dark, washed in 0.5% metabisulfate twice and then washed in water until the gel destained. The gel was then photographed. As seen in Figure 1, results showed red staining of the glycoproteins.

Bottom Line: The gel was first stained with periodic acid Schiff (PAS), a mucin histochemical stain followed by a secondary protein staining with Simply Blue Safestain (Invitrogen).Visual scoring (based on the presence of mucins) gave a sensitivity of 95%, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 88% for prediction of mucinous histology.One dimensional SDS polyacrylamide gel electrophoresis of pancreatic cyst fluid, followed by mucin (PAS) and protein (Simply Blue Safestain) staining, provides a means of concentrating and visualizing mucins, which allows the accurate differentiation of mucinous from non-mucinous histology in pancreatic cysts.

View Article: PubMed Central - PubMed

Affiliation: Departments of Surgery, Essentia Health System/Duluth Clinic, University of Minnesota Medical School, Duluth, MN USA.

ABSTRACT

Background: Pancreatic cysts are being increasingly identified in patients. Mucinous cysts have malignant potential whereas non-mucinous cysts do not. Distinguishing potentially malignant cysts from harmless ones by the characterization of cyst fluid contents remains a difficult problem. This study was undertaken to determine whether cyst fluid mucin glycoprotein analysis could differentiate mucinous from non-mucinous pancreatic cysts.

Methods: Cyst fluid from 28 patients who underwent resection of a pancreatic cyst was used for the study. In each case the type of cyst was histologically identified. One dimensional SDS polyacrylamide gel electrophoresis (1D-SDS PAGE) was performed on cyst fluid samples. For the detection of the separated proteins, we employed a novel dual staining technique. The gel was first stained with periodic acid Schiff (PAS), a mucin histochemical stain followed by a secondary protein staining with Simply Blue Safestain (Invitrogen).

Results: Visual scoring (based on the presence of mucins) gave a sensitivity of 95%, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 88% for prediction of mucinous histology.

Conclusions: One dimensional SDS polyacrylamide gel electrophoresis of pancreatic cyst fluid, followed by mucin (PAS) and protein (Simply Blue Safestain) staining, provides a means of concentrating and visualizing mucins, which allows the accurate differentiation of mucinous from non-mucinous histology in pancreatic cysts.

No MeSH data available.


Related in: MedlinePlus