TY - JOUR A1 - Gericke, Beate A1 - Koebnick, Corinna A1 - Reimann, Manja A1 - Forterre, Simone A1 - Zunft, Hans-Joachim Franz A1 - Schweigert, Florian J. T1 - Influence of hormone replacement therapy on proteomic pattern in serum of postmenopausal women N2 - Objectives: Proteomics approaches to cardiovascular biology and disease hold the promise of identifying specific proteins and peptides or modification thereof to assist in the identification of novel biomarkers. Method: By using surface-enhanced laser desorption and ionization time of flight mass spectroscopy (SELDI-TOF-MS) serum peptide and protein patterns were detected enabling to discriminate between postmenopausal women with and without hormone replacement therapy (HRT). Results: Serum of 13 HRT and 27 control subjects was analyzed and 42 peptides and proteins could be tentatively identified based on their molecular weight and binding characteristics on the chip surface. By using decision tree-based Biomarker Patterns (TM) Software classification and regression analysis a discriminatory function was developed allowing to distinguish between HRT women and controls correctly and, thus, yielding a sensitivity of 100% and a specificity of 100%. The results show that peptide and protein patterns have the potential to deliver novel biomarkers as well as pinpointing targets for improved treatment. The biomarkers obtained represent a promising tool to discriminate between HRT users and non-users. Conclusion: According to a tentative identification of the markers by their molecular weight and binding characteristics, most of them appear to be part of the inflammation induced acute-phase response. (c) 2004 Elsevier Ireland Ltd. All rights reserved Y1 - 2004 ER - TY - JOUR A1 - Schweigert, Florian J. A1 - Gericke, Beate A1 - Mothes, Ralf T1 - Lebensmittelanalytik mittels SELDI-TOF-MS Y1 - 2003 ER - TY - JOUR A1 - Gericke, Beate A1 - Raila, Jens A1 - Sehouli, Jalid A1 - Haebel, Sophie A1 - Konsgen, D. A1 - Mustea, A. A1 - Schweigert, Florian J. T1 - Microheterogeneity of transthyretin in serum and ascitic fluid of ovarian cancer patients N2 - Background: Transthyretin (TTR), a traditional biomarker for nutritional and inflammatory status exists in different molecular variants of yet unknown importance. A truncated form of TTR has recently been described to be part of a set of biomarkers for the diagnosis of ovarian cancer. The main aim of the study was therefore to characterize differences in microheterogeneity between ascitic fluid and plasma of women affected with ovarian cancer and to evaluate the tumor site as the possible source of TTR. Methods: Subjects were 48 women with primary invasive epithelial ovarian cancer or recurrent ovarian carcinoma. The control group consisted of 20 postmenopausal women. TTR and retinol-binding protein (RBP) levels were measured by enzyme-linked immunoassay ( ELISA) and C-reactive protein (CRP) levels by a high- sensitivity latex particle turbidimetric assay. The molecular heterogeneity of TTR was analysed using immunoprecipitation and matrix-associated laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Presence of TTR in tumor tissue was determined with indirect peroxidase immunostaining. Results: TTR and RBP (mu g/ml) levels in serum were 148.5 +/- 96.7 and 22.5 +/- 14.8 in affected women compared to 363.3 +/- 105.5 and 55.8 +/- 9.3 in healthy postmenopausal women ( p < 0.01). In ascitic fluid, levels were 1.02 +/- 0.24 and 4.63 +/- 1.57 mu g/ml, respectively. The mean levels of TTR and RBP in serum showed a tendency to decrease with the severity of the disease and were lower in affected women whose CRP levels were > 40 mg/ml ( p = 0.08 for TTR; p < 0.05 for RBP). No differences in TTR microheterogeneity were observed between TTR isolated from serum of affected and healthy women or from ascitic fluid. TTR occurred rather consistently in four variants. Mass signals were at 13758 +/- 7, 13876 +/- 13 ( greatest intensity), 13924 +/- 21 and 14062 +/- 24 Da, representing native, S-cysteinylated, S-cysteinglycinylated and glutathionylated TTR, respectively. Serum of healthy and affected women as well as ascitic fluid contained the truncated fragment of TTR ( 12828 +/- 11 Da). No immunoreactive TTR was observed in the tumor sites. Conclusion: The severity of the cancer associated catabolism as well as the inflammation status affect serum TTR and RBP levels. Neither TTR nor its truncated form originates from tumor tissue and its occurrence in ascites may well reflect the filtration from blood into ascitic fluid Y1 - 2005 SN - 1471-2407 ER - TY - JOUR A1 - Schweigert, Florian J. A1 - Gericke, Beate A1 - Wolfram, Wiebke A1 - Kaisers, Udo A1 - Dudenhausen, Joachim W. T1 - Peptide and protein profiles in serum and follicular fluid of women undergoing IVF JF - Human reproduction N2 - BACKGROUND: Proteins and peptides in human follicular fluid originate from plasma or are produced by follicular structures. Compositional changes reflect oocyte maturation and can be used as diagnostic markers. The aim of the study was to determine protein and peptide profiles in paired serum and follicular fluid samples from women undergoing IVF. METHODS: Surface-enhanced laser desorption and ionization-time of flight-mass spectrometry (SELDI-TOF-MS) was used to obtain characteristic protein pattern. RESULTS: One hundred and eighty-six individual MS signals were obtained from a combination of enrichment on strong anion exchanger (110), weak cation exchanger (52) and normal phase surfaces (24). On the basis of molecular masses, isoelectric points and immunoreactivety, four signals were identified as haptoglobin (alpha(1)- and alpha(2)-chain), haptoglobin 1 and transthyretin (TTR). Immunological and MS characteristics of the TTR : retinol-binding protein (RBP) transport complex revealed no microheterogeneity differences between serum and follicular fluid. Discriminatory patterns arising from decision-tree-based classification and regression analysis distinguished between serum and follicular fluid with a sensitivity and specificity of 100%. CONCLUSIONS: Quantitative and qualitative differences indicate selective transport processes rather than mere filtration across the blood-follicle barrier. Identified proteins as well as characteristic peptide and/or protein signatures might emerge as potential candidates for diagnostic markers of follicle and/or oocyte maturation and thus oocyte quality. KW - human follicular fluid KW - peptide KW - protein KW - proteome KW - serum Y1 - 2006 U6 - https://doi.org/10.1093/humrep/del257 SN - 0268-1161 VL - 21 IS - 11 SP - 2960 EP - 2968 PB - Univ. Press CY - Oxford ER -