scholarly journals Proteome Analysis of Cultured Fibroblasts from Type 1 Diabetic Patients and Normal Subjects

2006 ◽  
Vol 91 (9) ◽  
pp. 3507-3514 ◽  
Author(s):  
Lucia Puricelli ◽  
Elisabetta Iori ◽  
Renato Millioni ◽  
Giorgio Arrigoni ◽  
Peter James ◽  
...  
Keyword(s):  
Human Skin ◽  
Proteome Analysis ◽  
Normal Subjects ◽  
Skin Fibroblasts ◽  
Structural Proteins ◽  
Diabetic Patients ◽  
Human Skin Fibroblasts ◽  
Cultured Fibroblasts ◽  
Type 1 Diabetic Patients

Abstract Context: Protein profiling of diabetic tissues could provide useful biomarkers for early diagnosis, therapeutic targets, and disease response markers. Cultured fibroblasts are a useful in vitro model for proteome analysis and study of the molecular mechanisms involved in diabetes. Objective: The objective of the study was to isolate and characterize the proteins of cultured fibroblasts, obtained by skin biopsy, from long-term type 1 diabetic patients without complications and age- and sex-matched normal subjects as controls. Design: Proteins were separated by two-dimensional electrophoresis (2-DE), and the gel images were qualitatively and quantitatively analyzed. Protein identification was performed by matrix-assisted laser desorption/ionization mass spectrometry. Results: Reproducible protein maps of fibroblasts from diabetic and healthy subjects were obtained. A total of 125 protein spots were isolated and identified, among them 27 proteins not previously reported in published human fibroblast 2-DE maps, including 20 proteins never reported previously in the literature in human skin fibroblasts. Quantitative analyses revealed six protein spots differentially expressed in the fibroblasts from the diabetic vs. the control subjects (P < 0.05), representing glycolytic enzymes and structural proteins. An increase of triosephosphate I isomerase of two splice isoforms of pyruvate kinase and α-actinin 4 and a decrease of tubulin-β2 and splice isoform 2 of tropomyosin β-chain were detected. Conclusions: We generated 2-DE reference maps of the proteome of human skin fibroblasts from both normal and uncomplicated type 1 diabetic patients. Differences in glycolytic enzymes and structural proteins were found. The functional implications of the identified proteins are discussed.

4-Methylumbelliferone induces the expression of membrane type 1-matrix metalloproteinase in cultured human skin fibroblasts

2002 ◽  
Vol 298 (5) ◽  
pp. 646-650 ◽  
Author(s):  
Toshiya Nakamura ◽  
Takashi Ishikawa ◽  
Naoki Nanashima ◽  
Tomisato Miura ◽  
Hiroyuki Nozaka ◽  
...  
Keyword(s):  
Matrix Metalloproteinase ◽  
Human Skin ◽  
Skin Fibroblasts ◽  
Human Skin Fibroblasts ◽  
Membrane Type

Insulin resistance is localized to skeletal but not heart muscle in type 1 diabetes

1993 ◽  
Vol 264 (5) ◽  
pp. E756-E762 ◽  
Author(s):  
P. Nuutila ◽  
J. Knuuti ◽  
U. Ruotsalainen ◽  
V. A. Koivisto ◽  
E. Eronen ◽  
...  
Keyword(s):  
Insulin Resistance ◽  
Glucose Uptake ◽  
Compartment Model ◽  
Normal Subjects ◽  
Whole Body ◽  
Diabetic Patients ◽  
Uptake Rates ◽  
Arm Muscles ◽  
Type 1 Diabetic Patients

To determine the tissue localization of insulin resistance in type 1 diabetic patients, whole body and regional glucose uptake rates were determined under euglycemic hyperinsulinemic conditions. Leg, arm, and heart glucose uptake rates were measured using positron emission tomography-derived 2-deoxy-2-[18F]-fluoro-D-glucose kinetics and the three-compartment model described by Sokoloff et al. (L. Sokoloff, M. Reivich, C. Kennedy, M.C. DesRosiers, C.S. Patlak, K.D. Pettigrew, O. Sakurada, and M. Shinohara. J. Neurochem. 28: 897–916, 1977) in eight type 1 diabetic patients and eight matched normal subjects. Whole body glucose uptake was quantitated by the euglycemic insulin clamp technique. Whole body glucose uptake was approximately 31% lower in the diabetic patients (P < 0.01) than in the normal subjects, thus confirming the presence of whole body insulin resistance. The rate of glucose uptake was approximately 45% lower in leg muscle when measured in the femoral region (55 +/- 7 vs. 102 +/- 13 mumol.kg muscle-1.min-1, diabetic patients vs. normal subjects, P < 0.05) and approximately 27% lower in the arm muscles (66 +/- 4 vs. 90 +/- 13 mumol.kg muscle-1.min-1, respectively, P < 0.05), whereas no difference was observed in heart glucose uptake [789 +/- 80 vs. 763 +/- 58 mumol.kg muscle-1.min-1 not significant (NS)]. Whole body glucose uptake correlated with glucose uptake in femoral (r = 0.93, P < 0.005) and arm muscles (r = 0.66, P < 0.05) but not with glucose uptake in the heart (r = 0.04, NS). We conclude that insulin resistance in type 1 diabetic patients is localized to skeletal muscle, whereas heart glucose uptake is unaffected.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Glomerular cell number in normal subjects and in type 1 diabetic patients

2001 ◽  
Vol 59 (6) ◽  
pp. 2104-2113 ◽  
Author(s):  
Michael W. Steffes ◽  
Derek Schmidt ◽  
Rebecca Mccrery ◽  
John M. Basgen
Keyword(s):  
Normal Subjects ◽  
Cell Number ◽  
Diabetic Patients ◽  
Glomerular Cell ◽  
Type 1 Diabetic Patients

scholarly journals In situ protein Kinase C activity is increased in cultured fibroblasts from Type 1 diabetic patients with nephropathy

Diabetologia ◽  
2003 ◽  
Vol 46 (4) ◽  
pp. 524-530 ◽  
Author(s):  
E. Iori ◽  
M. C. Marescotti ◽  
M. Vedovato ◽  
G. Ceolotto ◽  
A. Avogaro ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Diabetic Patients ◽  
Cultured Fibroblasts ◽  
Kinase C ◽  
Protein Kinase C Activity ◽  
Type 1 Diabetic Patients

Circulating growth hormone forms in Type 1 diabetic subjects: comparison with normal subjects and acromegalics

1986 ◽  
Vol 112 (4) ◽  
pp. 547-551 ◽  
Author(s):  
L. A. MacFarlane ◽  
Susan Stafford ◽  
A. D. Wright
Keyword(s):  
Growth Hormone ◽  
Normal Subjects ◽  
Molecular Forms ◽  
Diabetic Patients ◽  
Strongly Correlated ◽  
Gh Secretion ◽  
Main Component ◽  
Type 1 Diabetic Patients ◽  
Serum Gh

Abstract. The molecular forms of growth hormone (GH) in serum from 18 Type 1 diabetic patients with poor metabolic control were analysed using sephadex G-100 chromatography. The profiles obtained were compared with those from normal subjects whose GH secretion was stimulated by exercise and hypoglycaemia and eight acromegalic patients. In the three groups three distinct GH forms were found: little (monomeric), big and big-big-GH. Samples from normal subjects contained 45% little-GH which was less than samples from the diabetics and acromegalics (53% and 65%, respectively, P <0.01). Further samples from normal subjects after the onset of hypoglycaemia showed an increase in little-GH. In the three groups, the higher the proportion of little-GH, the lower the proportion of big-big-GH, while the proportion of big-GH remained similar. In the acromegalics the proportion of little-GH was strongly correlated with the log concentration of serum GH. As little-GH is cleared from the circulation quicker than the larger forms these data indicate that the main component of the frequent surges of GH secretion in poorly controlled Type 1 diabetic subjects is little-GH (monomeric forms). The sustained release of GH found in acromegaly is composed largely of monomeric forms.

Effects of CK2 inhibition in cultured fibroblasts from Type 1 Diabetic patients with or without nephropathy

2015 ◽  
Vol 33 (4) ◽  
pp. 259-266 ◽  
Author(s):  
Elisabetta Iori ◽  
Maria Ruzzene ◽  
Sofia Zanin ◽  
Stefano Sbrignadello ◽  
Lorenzo Alberto Pinna ◽  
...  
Keyword(s):  
Diabetic Patients ◽  
Cultured Fibroblasts ◽  
Type 1 Diabetic Patients

scholarly journals Biosynthesis and release of glycoproteins by human skin fibroblasts in culture

1977 ◽  
Vol 168 (1) ◽  
pp. 91-103 ◽  
Author(s):  
Christopher H. J. Sear ◽  
Michael E. Grant ◽  
David S. Jackson
Keyword(s):  
Human Skin ◽  
De Novo ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Skin Fibroblasts ◽  
Human Skin Fibroblasts ◽  
Cultured Fibroblasts ◽  
Cscl Density Gradient ◽  
Before And After

1. Confluent human skin fibroblasts maintained in a chemically defined medium incorporate l-[1-3H]fucose in a linear manner with time into non-diffusible macromolecules for up to 48h. Chromatographic analysis demonstrated that virtually all the macromolecule-associated3H was present as [3H]fucose. 2. Equilibrium CsCl-density-gradient centrifugation established that [3H]fucose-labelled macromolecules released into the medium were predominantly glycoproteins. Confirmation of this finding was provided by molecular-size analyses of the [3H]fucose-labelled material before and after trypsin digestion. 3. The [3H]fucose-labelled glycoproteins released into fibroblast culture medium were analysed by gel-filtration chromatography and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. These techniques demonstrated that the major fucosylated glycoprotein had an apparent mol.wt. of 230000–250000; several minor labelled species were also detected. 4. Dual-labelling experiments with [3H]fucose and14C-labelled amino acids indicated that the major fucosylated glycoprotein was synthesized de novo by cultured fibroblasts. The non-collagenous nature of this glycoprotein was established by three independent methods. 5. Gel-filtration analysis before and after reduction with dithiothreitol showed that the major glycoprotein occurs as a disulphide-bonded dimer when analysed under denaturing conditions. Further experiments demonstrated that this glycoprotein was the predominant labelled species released into the medium when fibroblasts were incubated with [35S]cysteine. 6. The relationship between the major fucosylated glycoprotein and a glycoprotein, or group of glycoproteins, variously known as fibronectin, LETS protein, cell-surface protein etc., is discussed.

Sign in / Sign up

Export Citation Format

Share Document