A high performance gel filtration chromatography method for γ-glutamyltransferase fraction analysis

Franzini, Maria; Bramanti, Emilia; Ottaviano, Virginia; Ghiri, Emiliano; Scatena, Fabrizio; Barsacchi, Renata; Pompella, Alfonso; Donato, L.; Emdin, Michele; Paolicchi, Aldo

doi:10.1016/j.ab.2007.10.025

Cited by 58 publications

(67 citation statements)

References 26 publications

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“…Fractional GGT analysis was performed as previously described [27,28] using an FPLC (fast protein liquid chromatography) system (AKTA purifier, GE Healthcare Europe, Milan, Italy) equipped with a gel-filtration column (Superose 6 HR 10/300 GL, GE Healthcare Europe) and a fluorescence detector (Jasco FP-2020, Jasco Europe, Lecco, Italy). Plaque homogenates and plasma samples (0.02 ml) were injected in the gel-filtration column, after being filtered with a 0.45 mm polyvinylidene fluoride Millipore filter.…”

Section: Fractional Ggt Analysismentioning

confidence: 99%

“…The GGT activity has been hypothesized to exert a direct pro-oxidant activity, eliciting the production of reactive oxygen species from cystenylglycine (CysGly) moieties via its catalytic activity on extracellular glutathione [23e25], the occurrence of GGT-mediated redox reactions having been demonstrated within the plaque [26]. Recently, four plasma GGT fractions have been identified by molecular size exclusion chromatography, i.e., b-GGT, m-GGT and s-GGT (that very likely represent >2000, 940 and 140 kDa, respectively, lipoproteinbound molecular masses) plus the 70 kDa free fraction f-GGT [27,28]; only the highest molecular weight fraction b-GGT, has been found within atherosclerotic plaques by our group [26].…”

Section: Introductionmentioning

confidence: 99%

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b-Gamma-glutamyltransferase activity in human vulnerable carotid plaques

Pucci¹,

Franzini

Matteucci

et al. 2014

Atherosclerosis

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a b s t r a c tObjective: The atherosclerotic plaque that is vulnerable to rupture and to superimposed thrombosis is mainly represented by a thin-cap fibroatheroma with or without ulceration/thrombosis and inflammatory infiltrates. Total serum gamma-glutamyltransferase (GGT) activity is an independent predictor for cardiovascular events. Four GGT fractions have been identified in plasma and only one of them (b-GGT) in atherosclerotic plaques, but the possible role of GGT in plaque pathophysiology has not been assessed yet. We investigated the relationships between plaque b-GGT activity and the histological features of plaque vulnerability. Methods and results: Plaque GGT activity was investigated in 65 patients undergoing carotid endarterectomy; plaques were histologically characterized and immunostained for GGT. Intra-plaque total and fractional GGT activity was determined by a cost-effective test of molecular size exclusion chromatography, and compared with histological markers of plaque vulnerability. Plaque cholesterol content was also measured by chromatography. b-GGT was the only fraction detected within the atherosclerotic plaques and intra-plaque b-GGT activity correlated to plaque cholesterol content (r ¼ 0.667, P < 0.0001), plasma b-GGT and f-GGT fractions (r ¼ 0.249; r ¼ 0.298, both P < 0.05). Higher b-GGT activity was found in thin-cap fibroatheromas and it was associated to histological markers of vulnerable plaques, i.e., larger necrotic areas, greater macrophage infiltration and higher cholesterol content (P < 0.05). Conclusions: intra-plaque b-GGT activity correlates with the histological markers of vulnerable plaque and with plasma b-GGT in human carotid atherosclerosis; these data support the possible role of b-GGT in clinically significant atherosclerotic disease.

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Section: Fractional Ggt Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

b-Gamma-glutamyltransferase activity in human vulnerable carotid plaques

Pucci¹,

Franzini

Matteucci

et al. 2014

Atherosclerosis

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“…Circulating GGT is supposed to originate mostly from the liver (12,13) and is influenced by genetic and environmental factors (14). Based on gel filtration chromatography, researchers from the Pisa University have identified 4 GGT fractions with different molecular weight in subjects of both sexes: big (b-GGT), medium (m-GGT), small (s-GGT) and free (f-GGT) (15). Recent studies have suggested that b-GGT consists of membrane microvesicles and it may serve as a precursor for smaller fractions (m-GGT and s-GGT) whereas f-GGT represents free soluble form of the enzyme (4).…”

Section: Figurementioning

confidence: 99%

Gamma-glutamyl transferase and cardiovascular disease

Ndrepepa¹,

Kastrati²

2016

Ann. Transl. Med.

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Gamma-glutamyl transferase (GGT) is an enzyme located on the external surface of cellular membranes. GGT contributes in maintaining the physiological concentrations of cytoplasmic glutathione and cellular defense against oxidative stress via cleavage of extracellular glutathione and increased availability of amino acids for its intracellular synthesis. Increased GGT activity is a marker of antioxidant inadequacy and increased oxidative stress. Ample evidence suggests that elevated GGT activity is associated with increased risk of cardiovascular disease (CVD) such as coronary heart disease (CHD), stroke, arterial hypertension, heart failure, cardiac arrhythmias and all-cause and CVD-related mortality. The evidence is weaker for an association between elevated GGT activity and acute ischemic events and myocardial infarction. The risk for CVD or CVD-related mortality mediated by GGT may be explained by the close correlation of GGT with conventional CVD risk factors and various comorbidities, particularly non-alcoholic fatty liver disease, alcohol consumption, oxidative stress, metabolic syndrome, insulin resistance and systemic inflammation. The finding of GGT activity in atherosclerotic plaques and correlation of intra-plaque GGT activity with histological indexes of plaque instability may suggest a participation of GGT in the pathophysiology of CVD, particularly atherosclerosis. However, whether GGT has a direct role in the pathophysiology of CVD or it is an epiphenomenon of coexisting CVD risk factors or comorbidities remains unknown and Hill's criteria of causality relationship between GGT and CVD are not fulfilled. The exploration whether GGT provides prognostic information on top of the information provided by known cardiovascular risk factors regarding the CVD or CVD-related outcome and exploration of molecular mechanisms of GGT involvement in the pathophysiology of CVD and eventual use of interventions to reduce circulating GGT activity remain a duty of future studies.

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“…We had exclude patients who were with liver disease and intestinal obstruction at the first time presented in our cancer center in order to limit the bias (He et al, 2013;He et al, 2013). Franzini had reported that there were at least four kinds of GGT, and b-GGT was produced other than liver (Franzini et al, 2008;Franzini et al, 2009), thus the subtypes of GGT may improve the specificity and severe as promising candidates in the further.…”

Section: Discussionmentioning

confidence: 99%

“…γ-glutamyl transpeptidase (GGT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) which were included in biochemical test, were also showed as prognostic factors (Saif et al, 2005;Koukourakis et al, 2006;Pompella et al, 2006;Franzini et al, 2008;Franzini et al, 2009;Xie et al, 2014). However, previous studies had several drawbacks.…”

Section: Prognostic Model Built On Blood-based Biomarkers In Patientsmentioning

confidence: 99%

Prognostic Model Built on Blood-based Biomarkers in Patients with Metastatic Colorectal Cancer

Jiang²,

Yin

et al. 2014

Asian Pacific Journal of Cancer Prevention

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Background: We had previously showed that the neutrophil lymphocyte ratio (NLR), γ-glutamyl transpeptidase (GGT) and carcinoembryonic antigen (CEA) are prognostic factors for metastatic colorectal cancer (mCRC) patients. In this study we developed a prognostic model based on these three indices. Materials and Methods: A total of 243 patients who were initially diagnosed as mCRC between 2005 and 2010 in the Sun Yat-sen University Cancer Center were studied. The endpoint was overall survival (OS). Results: NLR>3, elevated GGT and elevated CEA were confirmed as independent risk factors which could predict poor prognosis. Patients could be divided into three groups according to the number of risk factors they had. Those with two or three were defined as the high risk group, individuals with one risk factor as the modest risk group and patients without risk factor as the low risk group. The OS values for these three groups were 16.2 months (2.80~68.8), 24.2 months (4.07~79.0), and 37.2 months (12.6~87.8), respectively (p<0.001). Conclusions: We developed a simple but useful model based on NLR, GGT and CEA to provide prognostic information to clinical practice in highly selected mCRC patients. Further prospective and multi-center studies are warranted to test our model.

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A high performance gel filtration chromatography method for γ-glutamyltransferase fraction analysis

Cited by 58 publications

References 26 publications

b-Gamma-glutamyltransferase activity in human vulnerable carotid plaques

b-Gamma-glutamyltransferase activity in human vulnerable carotid plaques

Gamma-glutamyl transferase and cardiovascular disease

Prognostic Model Built on Blood-based Biomarkers in Patients with Metastatic Colorectal Cancer

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