Cíl studie: Zavedení nové techniky, kapalinové chromatografie ve spojení s tandemovou hmotnostní spektrometrií do rutinního stanovení hladiny celkového homocysteinu v plazmě. Materiál a metoda: Metodou kapalinové chromatografie ve spojení s tandemovou hmotnostní spektrometrií byly analyzovány vzorky pacientů zaslané do laboratoří Onkologického Centra J. G. Mendela. Výsledky: V pozitivním multiple reaction monitoring módu byl monitorován ion vzniklý rozpadem fragmentu m/z 135,9 → 89,8 pro homocystein a m/z 140 → 90 pro interní standard, homocystein-d4. Vyvinutá LC/MS/MS metoda má následující parametry: retenční čas homocysteinu i homocystinu-d8 1,4 minuty, mez stanovitelnosti 0,5 μmol/l, mez detekce 0,1 μmol/l. Opakovatelnost byla stanovena v sérii 20 měření analýzou 2 kontrolních vzorků, na hladině 8,3 μmol/l byl variační koeficient 0,12 %, na hladině 18,2 μmol/l byla hodnota variačního koeficientu 0,71 %. Závěr: Byla zavedena vysoce selektivní, specifická a časově nenáročná technika vhodná pro rutinní stanovení hladiny celkového homocysteinu v plazmě., Objective: To develop a new LC/MS/MS - based method for routine measurement of total homocysteine in plasma. Material and Method: Patient samples from laboratories of Cancer Centre of J. G. Mendel were analyzed by liquid chromatography with tandem mass spectrometry. Results: Ion transitions m/z 135.9 → 89.8 for homocysteine and m/z 140 → 90 for internal standard – homocysteine-d4 were monitored in the positive multiple reaction-monitoring mode. Our newly developed LC/MS/MS method has the following characteristics: retention time of homocysteine and homocystine-d8 is 1.4 min, limit of quantitation 0.5 μmol/l and limit of detection 0.1 μmol/l. Repeatability was determined by analyzing 2 plasma samples, the variation coefficients obtained were 0.12 % at concentration of 8.3 μmol/l and 0.71 % at concentration of 18.2 μmol/l. Conclusion: The selective, specific and rapid method suitable for routine determination of total homocysteine in plasma was developed., Soňa Humplíková, J. Minář, M. Kučerová, and Lit. 18
Monoclonal antibody-based treatment of cancer has been established as one of the most successful therapeutic strategies for both hematologic malignancies and solid tumors. In addition to targeting cancer antigens antibodies can also modulate immunological pathways that are critical to immune surveillance. Antibody therapy directed against several negative immunologic regulators (checkpoints) is demonstrating significant success in the past few years. Immune checkpoint inhibitors, ipilimumab,
pembrolizumab and nivolumab, have shown significant clinical benefit in several malignancies and are already approved for advanced melanoma and squamous NSCLC. Based on their mechanism of action, these agents can exert toxicities that are unlike conventional cytotoxic chemotherapy, whose nature is close to autoimmune diseases -
immune related adverse events (irAEs). In this review we focus on the spectrum of irAEs associated with immune checkpoint antibodies, discussing the pharmacological treatment strategy and possible clinical impact.