Reviewer's report 
 Oxidised and reduced glutathione and related thiols in human gallbladder bile. Title: 
 1  14 September 2006 Version: Date: 
 Ignazio Grattagliano Reviewer: 
Reviewer's report: 
General 
------------------------------------------------------------------------------- 
Major Compulsory Revisions (that the author must respond to before a decision on publication can be reached 
The authors have attempted to assess and define several different thiols concentrations in human GB bile. 
Although the method for the measurement was correct, they fail in their question, patients selection and 
data interpretation. 
Specific Comments 
Abstract 
- Objective: the first sentence is not impressive and should be removed; 
- Results and conclusions: mention and comparison with rodent bile composition is inappropriate. 
Introduction 
- Generally, abbreviations used for reduced and oxidized glutathione are GSH and GSSG, respectively; - The scientific and clinical relevance to measure biliary thiols is to have more information on the 
composition of this biological fluid in humans and eventually to know if its manipulation by medications 
could represent any therapeutic option in biliary disease. Bile thiols may have importance for protein and lipid maintenance in the reduced status. The indication of the hepatic thiol status and degree of chemical/oxidative stress is conversely an unlikely application. 
- No mention was done on the possible GB (mucosal secretion) origin of at least part of GB bile free thiols. 
It is difficult to believe that GB bile thiols just reflect canalicular bile composition. Bile generally remains in GB at least for some hours, therefore it can be largely affected by GB motility and inflammatory status. Patients 
- More clinical information on patients need: were they under cholestatis? According to the reported 
conditions, it is highly probable that they were cholestatic; therefore, the results do not reflect human bile thiols in a healthy status which was the main objective of the study, and could explain the very high variety found; 
- Patients with GB stones were not classified according to the type and number of stones. Number, type and 
dimension of stones are known to be related with protein concentration and cholesterol crystallization time. 
Methods - Cause of the fast hydrolization of biliary GSH due to the high presence of gGT in the biliary tree and GB, 
freshly removed bile should be immediately treated with gGT inhibitors, such as serin-borate. 
Results - The higher levels of GSH and GSSG found in patients with cholecystitis supports the hypothesis that at least part of GB GSH origin directly from GB tissues (vessels and inflamed mucosa). 
Discussion 
- Patient population was too eterogeneous; no comparison can be made with a more homogeneous population of rodents (mice and guinea pig in particular that have the GB). Conclusions 
- The first and second points are not appropriate; possible explanation for the third is above sugg 
------------------------------------------------------------------------------- 
Minor Essential Revisions (such as missing labels on figures, or the wrong use of a term, which the author 
can be trusted to correct) 
------------------------------------------------------------------------------- 
Discretionary Revisions (which the author can choose to ignore) 
 Reject because scientifically unsound What next?: 
 An article of limited interest Level of interest: 
 Acceptable Quality of written English: 
 No Statistical review: 
Declaration of competing interests: 
I declare I have no competing interest 
Reviewer's report 
 Oxidised and reduced glutathione and related thiols in human gallbladder bile. Title: 
 1  14 September 2006 Version: Date: 
 Ignazio Grattagliano Reviewer: 
Reviewer's report: 
General 
------------------------------------------------------------------------------- 
Major Compulsory Revisions (that the author must respond to before a decision on publication can be reached 
The authors have attempted to assess and define several different thiols concentrations in human GB bile. 
Although the method for the measurement was correct, they fail in their question, patients selection and 
data interpretation. 
Specific Comments 
Abstract 
- Objective: the first sentence is not impressive and should be removed; 
- Results and conclusions: mention and comparison with rodent bile composition is inappropriate. 
Introduction 
- Generally, abbreviations used for reduced and oxidized glutathione are GSH and GSSG, respectively; - The scientific and clinical relevance to measure biliary thiols is to have more information on the 
composition of this biological fluid in humans and eventually to know if its manipulation by medications 
could represent any therapeutic option in biliary disease. Bile thiols may have importance for protein and lipid maintenance in the reduced status. The indication of the hepatic thiol status and degree of chemical/oxidative stress is conversely an unlikely application. 
- No mention was done on the possible GB (mucosal secretion) origin of at least part of GB bile free thiols. 
It is difficult to believe that GB bile thiols just reflect canalicular bile composition. Bile generally remains in GB at least for some hours, therefore it can be largely affected by GB motility and inflammatory status. Patients 
- More clinical information on patients need: were they under cholestatis? According to the reported 
conditions, it is highly probable that they were cholestatic; therefore, the results do not reflect human bile thiols in a healthy status which was the main objective of the study, and could explain the very high variety found; 
- Patients with GB stones were not classified according to the type and number of stones. Number, type and 
dimension of stones are known to be related with protein concentration and cholesterol crystallization time. 
Methods - Cause of the fast hydrolization of biliary GSH due to the high presence of gGT in the biliary tree and GB, 
freshly removed bile should be immediately treated with gGT inhibitors, such as serin-borate. 
Results - The higher levels of GSH and GSSG found in patients with cholecystitis supports the hypothesis that at least part of GB GSH origin directly from GB tissues (vessels and inflamed mucosa). 
Discussion 
- Patient population was too eterogeneous; no comparison can be made with a more homogeneous population of rodents (mice and guinea pig in particular that have the GB). Conclusions 
- The first and second points are not appropriate; possible explanation for the third is above sugg 
------------------------------------------------------------------------------- 
Minor Essential Revisions (such as missing labels on figures, or the wrong use of a term, which the author 
can be trusted to correct) 
------------------------------------------------------------------------------- 
Discretionary Revisions (which the author can choose to ignore) 
 Reject because scientifically unsound What next?: 
 An article of limited interest Level of interest: 
 Acceptable Quality of written English: 
 No Statistical review: 
Declaration of competing interests: 
I declare I have no competing interest 
Please review this paper.
user
Reviewer's report 
 Oxidised and reduced glutathione and related thiols in human gallbladder bile. Title: 
 1  14 September 2006 Version: Date: 
 Ignazio Grattagliano Reviewer: 
Reviewer's report: 
General 
------------------------------------------------------------------------------- 
Major Compulsory Revisions (that the author must respond to before a decision on publication can be reached 
The authors have attempted to assess and define several different thiols concentrations in human GB bile. 
Although the method for the measurement was correct, they fail in their question, patients selection and 
data interpretation. 
Specific Comments 
Abstract 
- Objective: the first sentence is not impressive and should be removed; 
- Results and conclusions: mention and comparison with rodent bile composition is inappropriate. 
Introduction 
- Generally, abbreviations used for reduced and oxidized glutathione are GSH and GSSG, respectively; - The scientific and clinical relevance to measure biliary thiols is to have more information on the 
composition of this biological fluid in humans and eventually to know if its manipulation by medications 
could represent any therapeutic option in biliary disease. Bile thiols may have importance for protein and lipid maintenance in the reduced status. The indication of the hepatic thiol status and degree of chemical/oxidative stress is conversely an unlikely application. 
- No mention was done on the possible GB (mucosal secretion) origin of at least part of GB bile free thiols. 
It is difficult to believe that GB bile thiols just reflect canalicular bile composition. Bile generally remains in GB at least for some hours, therefore it can be largely affected by GB motility and inflammatory status. Patients 
- More clinical information on patients need: were they under cholestatis? According to the reported 
conditions, it is highly probable that they were cholestatic; therefore, the results do not reflect human bile thiols in a healthy status which was the main objective of the study, and could explain the very high variety found; 
- Patients with GB stones were not classified according to the type and number of stones. Number, type and 
dimension of stones are known to be related with protein concentration and cholesterol crystallization time. 
Methods - Cause of the fast hydrolization of biliary GSH due to the high presence of gGT in the biliary tree and GB, 
freshly removed bile should be immediately treated with gGT inhibitors, such as serin-borate. 
Results - The higher levels of GSH and GSSG found in patients with cholecystitis supports the hypothesis that at least part of GB GSH origin directly from GB tissues (vessels and inflamed mucosa). 
Discussion 
- Patient population was too eterogeneous; no comparison can be made with a more homogeneous population of rodents (mice and guinea pig in particular that have the GB). Conclusions 
- The first and second points are not appropriate; possible explanation for the third is above sugg 
------------------------------------------------------------------------------- 
Minor Essential Revisions (such as missing labels on figures, or the wrong use of a term, which the author 
can be trusted to correct) 
------------------------------------------------------------------------------- 
Discretionary Revisions (which the author can choose to ignore) 
 Reject because scientifically unsound What next?: 
 An article of limited interest Level of interest: 
 Acceptable Quality of written English: 
 No Statistical review: 
Declaration of competing interests: 
I declare I have no competing interest 
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