Targeted On-line SPE-LC-MS/MS Assay for the Quantitation of 12 Apolipoproteins from Human Blood.

Laborious sample pretreatment of biological samples represents the most limiting factor for the translation of targeted proteomics assays from research to clinical routine. An optimized method for the simultaneous quantitation of 12 major apolipoproteins (apos) combining on-line SPE and fast LC-MS/MS analysis in 6.5 min total run time was developed, reducing the manual sample pretreatment time of 3 µL serum or plasma by 60%. Within-run and between-day imprecisions below 10 and 15% (n = 10) and high recovery rates (94-131%) were obtained applying the high-throughput setup. High-quality porcine trypsin was used, which outperformed cost-effective bovine trypsin regarding digestion efficiency. Comparisons with immunoassays and another LC-MS/MS assay demonstrated good correlation (Pearson's R: 0.81-0.98). Further, requirements on sample quality concerning sampling, processing, and long-term storage up to 1 year were investigated revealing significant influences of the applied sampling material and coagulant on quantitation results. Apo profiles of 1339 subjects of the LIFE-Adult-Study were associated with lifestyle and physiological parameters as well as establish parameters of lipid metabolism (e.g., triglycerides, cholesterol). Besides gender effects, most significant impact was seen regarding lipid-lowering medication. In conclusion, this novel highly standardized, high-throughput targeted proteomics assay utilizes a fast, simultaneous analysis of 12 apos from least sample amounts.

The rationale for radical cystectomy as primary therapy for T4 bladder cancer.

Treatment of T4 bladder cancer patients remains a clinical challenge. Conservative management is often insufficient regarding local control, neoadjuvant chemotherapy delays definite treatment while leading to increased therapy-associated morbidity and mortality during the course of the disease. Primary cystectomy has also been reported to be associated with a high complication rate and unsatisfactory clinical efficacy. Herein, we report postoperative outcome, including therapy-related complications, in 20 T4 bladder cancer patients subjected to primary cystectomy. Twenty patients underwent radical cystectomy for T4 bladder cancer. At the time of surgery, 8 patients had regional lymph node metastases. The median postoperative follow-up was 13 months for the whole group. Mean duration of postoperative hospitalization was 19 days. Ten patients received no intra- or postoperative blood transfusions, whereas an average number of 3 blood units were administered in the remaining cases. Major therapy-associated complications were paresthesia affecting the lower extremities (n = 3) as well as insignificant pulmonary embolism, enterocutaneous fistulation and acute renal failure in one patient, respectively. At the time of data evaluation, 11 patients were still alive after a follow-up of 20 months. Four patients >or=70 years at the time of cystectomy were still alive after 11, 22 and 31 months following surgery, respectively. The current data demonstrate primary cystectomy for T4 bladder cancer as a technically feasible approach that is associated with a tolerable therapy-related morbidity. Additionally, satisfying clinical outcome is observed even in a substantial number of elderly patients.