Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) methods for the therapeutic drug monitoring of cytotoxic anticancer drugs: An update.

In the era of precision medicine, there is increasing evidence that conventional cytotoxic agents may be suitable candidates for therapeutic drug monitoring (TDM)- guided drug dosage adjustments and patient's tailored personalization of non-selective chemotherapies. To that end, many liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) assays have been developed for the quantification of conventional cytotoxic anticancer chemotherapies, that have been comprehensively and critically reviewed. The use of stable isotopically labelled internal standards (IS) of cytotoxic drugs was strikingly uncommon, accounting for only 48 % of the methods found, although their use could possible to suitably circumvent patients' samples matrix effects variability. Furthermore, this approach would increase the reliability of cytotoxic drug quantification in highly multi-mediated cancer patients with complex fluctuating pathophysiological and clinical conditions. LC-MS/MS assays can accommodate multiplexed analyses of cytotoxic drugs with optimal selectivity and specificity as well as short analytical times and, when using stable-isotopically labelled IS for quantification, provide concentrations measurements with a high degree of certainty. However, there are still organisational, pharmacological, and medical constraints to tackle before TDM of cytotoxic drugs can be more largely adopted in the clinics for contributing to our ever-lasting quest to improve cancer treatment outcomes.

[A latent form of essential thrombocythemia presented as portal hypertension and associated with acquired von Willebrand syndrome].

A 32 year-old male patient was admitted to our hospital because of abdominal tumor. The examination on admission showed massive splenomegaly and esophageal varices although peripheral blood cell counts were within normal limits. Exploratory laparotomy was performed with the diagnosis of portal hypertension and revealed the multiple thrombus formations in the splenic vein and the extramedullary hematopoietic findings in the spleen by the microscopic examination. In vitro colony forming assay showed the formation of spontaneous erythroid colonies in cultures of progenitor cells (from peripheral blood mononuclear cells) in erythropoietin-poor medium. Increasing thrombocytosis was observed immediately after splenectomy, and hemorrhagic diathesis of nasal bleeding and gastrointestinal bleeding were also detected. The analysis of plasma von Willebrand factor (vWF) revealed the decrease of ristocetin cofactor activity and the lack of large multimeric components of vWF. These abnormal findings observed after splenectomy led to recovery through the administration of busulfan with the improvement of thrombocytosis. Accordingly, the course of the disease clearly indicated it to be the essential thrombocythemia represented as portal vein thrombosis and in latent form with normal cell counts in peripheral blood at the time of diagnosis, and subsequently, to develop into a full-blown form associated with acquired von Willebrand syndrome following splenectomy.

Homozygous deletion of INK4a/ARF genes and overexpression of bcl-2 in relation with poor prognosis in immunocompetent patients with primary central nervous system lymphoma of the diffuse large B-cell type.

Only a few reports have been published on molecular genetic alterations in primary central nervous system lymphomas (PCNSLs) of the diffuse large B-cell type and no reports have addressed the correlation between the genetic alterations and clinical course of the patients with this neoplasm. Thus, the molecular background of the PCNSL and its importance for the clinical course of the patients are still unclear. We investigated a series of 14 patients with PCNSL to determine structural alterations of the INK4a/ARF, MDM2, and TP53 genes, the status of bcl-2 and bcl-6 protein expression, and the clinical course of the patients (i.e. their survival time after diagnosis). No structural alterations of MDM2 and TP53 genes were found. Only INK4a/ARF genes whose expression affects both the p161NK4a-Rb and p14ARF-mdm2-p53 pathways in the regulation for cell cycle and apoptosis, showed an alteration of the homozygous deletions at a high frequency (nine of 14 patients: 64%). This specific alteration was not related with the bcl-6 expression, but a relation was shown with overexpression of the bcl-2 anti-apoptotic protein (p = 0.036, chi-square test), as well as a shorter patient survival (p = 0.044, Wilcoxon test). There was only a tendency, not a significant correlation, in which the patients with bcl-2 overexpression resulted in poor prognosis (p = 0.149). The present study is the first to suggest that the INK4a/ARF gene homozygous deletions and overexpression of the bcl-2 protein may be correlated with each other and together serve as important predictors for the prognosis of patients with PCNSL.