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Posttranscriptional Changes of Serum
Clinical and Prognostic Significance in
Patients With Cirrhosis

PPDS: dr. Reagan Paulus Rintar



such as antioxidant and scavenging activities. HSA presents other biological properties. binding and transport of many endogenous and exogenous substances.INTRODUCTION  Human serum albumin (HSA) is the most abundant plasma protein. and regulation of endothelial function and inflammatory response . and its oncotic power largely determines the fluid distribution in the different compartments of the body  Besides its oncotic capacity.

HSA is immersed in a stressful microenvironment as a result of the elevated serum concentration of proinflammatory and pro-oxidant substances . the occurrence of lowlevel posttranscriptional changes. and truncation involving the Cys-34 and other molecular sites. glycosylation. contributes to the microheterogeneity of the circulating molecule  In patients with advanced liver disease.INTRODUCTION  Physiological conditions. resulting from oxidation.

And. a significant damage of the molecule can be expected in these patients. which predominantly reflects the cobaltchelating activity at the Nterminal portion. the circulating level of ischemia modified albumin.6 is significantly increased in patients with acuteon-chronic liver failure .INTRODUCTION  Diabetes is present in approximately one third of patients with cirrhosis. besides the universal finding of hypoalbuminemia. additional stress can also derive from hyperglycemia. the reversible and irreversible oxidation of the Cys-34 residue occurs in advanced cirrhosis  Furthermore. indeed.

we assessed whether these HSA isoforms are associated with the severity of disease. . specific clinical features. and patient prognosis. we first analyzed the HAS posttranscriptional changes in a large cohort of patients with cirrhosis admitted to the hospital because of a clinical complication of the disease.INTRODUCTION  In the present study.  Then.

all the patients with cirrhosis admitted to our department  The diagnosis of cirrhosis was based on clinical. biochemical. and endoscopic features . ultrasound (US).PATIENTS AND METHODS  From July 2011 to March 2012.

.Exclusion criteria  Age under 18 years  Admission for a scheduled procedure  Hepatocellular carcinoma (HCC) exceeding the Milan criteria  Heart and respiratory failure  Organic renal diseases  Oncohematologic disorders  Protein-losing syndromes  Albumin infusion in the previous month  Ongoing immunosuppressive treatment.

and plasma was aliquoted into cryotubes. and stored at -80C until analysis . peripheral blood was withdrawn from the brachial vein into pyrogen-free tubes  Blood samples were immediately centrifuged at 3. and within 24 hours from admission for hospitalized patients.000xg for 10 minutes.Study Design  At the time of inclusion for outpatients with cirrhosis and controls.

Study Design  Posttranscriptional HSA molecular changes were identified and quantified by using a highperformance liquid chromatography/electrospray ionization mass spectrometry technique. .

Study Design  Clinical and biochemical parameters were also recorded and hospitalized  Patients were followed for up to 1 year. 7 HSA isoforms carrying one or more posttranscriptional changes were identified .

Statistical Analysis  Variables were expressed as mean and SD or frequencies according to their distribution.  Comparisons between categorical variables were made by means of the chi-square (x2) test. outpatients.  Differences in HSA isoform relative abundance between control subjects. and hospitalized patients with cirrhosis were assessed by one-way analysis of variance (ANOVA) with Bonferroni’s correction for multiple comparisons .

MELD and Child-Pugh scores.Statistical Analysis  The relationship between relative abundance of HSA isoforms and MELD and Child-Pughscores was evaluated with Spearman’s rho. and bacterial infection) in hospitalized patients was assessed by means of the Student t test  The association between HSA isoforms. and specific clinical complication (ascites. renal impairment. serum albumin concentration. and 1-year survival was assessed using Cox’s proportional hazard .  The univariate analysis of the association between HAS isoforms relative amount. age.

 The analyses were performed using SPSS Statistics 20. and values of P < 0.05 were considered statistically significant.Statistical Analysis  The best cutoffs of HSA isoforms significantly associated with patients survival and of serum albumin concentration were determined through receiver operating characteristic (ROC) curve analysis in order to plot survival curves using Kaplan-Meier’s method  All tests were two sided.0 software .

Clinical Characteristics of Patients With Cirrhosis at Study Enrollment .RESULT S Data are presented as mean ± SD or frequencies (%). †Prothrombin time Table 1.5 mg/dL. *Renal impairment = serum creatinine >1.

Fig. In addition to native HSA. seven HSA isoforms carrying the following structural alterations were detected: truncation of the last two amino acid residues at the Nterminal portion (HSA-DA). 1. sulfinylation of the Cys-34 residue (HSA1SO2H). and glycosylation (HSA1GLYC). Representative deconvoluted ESI-MS spectra from a control subject (A) and a patient with cirrhosis (B). truncation of the last amino acid residue at the C-terminal portion (HSA-L). Two additional HSA isoforms were generated from the combination of the cysteinylated with the N- . cysteinylation of the Cys-34 residue (HSA1CYS).






discriminating between isoforms whose molecular weight differs by a few Daltons .DISCUSSIONS  In this study. posttranscriptional structural changes of albumin in cirrhosis were assessed by HPLC/ESIMS for the first time.  This proteomic technique provides a spectrum of molecules with high resolution.

Kaplan-Meier’s survival curves for HSA1CYS-DA (A). native HSA (B).Fig. 3. and serum albumin concentration (C) in hospitalized patients with cirrhosis dichotomized according their best cut-off values .

the extent of many posttranscriptional molecular changes. which can also be detected at a low. Indeed. was significantly increased in patients . physiological level in healthy subjects.DISCUSSIONS  The first important finding of the present study is that the molecular structure of HSA is extensively altered in patients with cirrhosis.

involving several molecular sites and increasing in parallel with disease severity. . whereas the residual. native HSA isoform independently predicts patient survival.  Altered isoforms are independently associated with specific clinical complications. but also to the preservation of its structural integrity.” which implies that the global HSA function is related not only to its serum concentration.CONCLUSIONS  Extensive posttranscriptional changes of HSA. occur in patients with cirrhosis.  These findings support the concept of the “effective albumin concentration.