Serum mass spectrometry for treatment monitoring in patients with multiple myeloma receiving ARI0002h CAR T-cells.

Chimeric antigen receptor (CAR) T-cell therapies have increased the patients with relapsed/refractory multiple myeloma (RRMM) in whom standard electrophoretic techniques fail to detect the M-protein. Quantitative immunoprecipitation mass spectrometry (QIP-MS) can accurately measure serum M-protein with high sensitivity, and identify interferences caused by therapeutic monoclonal antibodies. Here, we investigate the outcome of QIP-MS in 33 patients treated with the academic BCMA-directed CAR T-cell ARI0002h (Cesnicabtagene Autoleucel). QIP-MS offered more detailed insights than serum immunofixation (sIFE), identifying glycosylated M-proteins and minor additional peaks. Moreover, the potential interferences owing to daratumumab or tocilizumab treatments were successfully detected. When analysing different assay platforms during patient's monitoring after ARI0002h administration, we observed that QIP-MS showed a high global concordance (78.8%) with sIFE, whereas it was only moderate (55.6%) with bone marrow (BM)-based next-generation flow cytometry (NGF). Furthermore, QIP-MS consistently demonstrated the lowest negativity rate across the different timepoints (27.3% vs. 60.0% in months 1 and 12, respectively). Patients with QIP-MS(+)/BM-based NGF(-) showed a non-significant shorter median progression free survival than those with QIP-MS(-)/BM-based NGF(-). In summary, we show the first experience to our knowledge demonstrating that QIP-MS could be particularly useful as a non-invasive technique when evaluating response after CAR T-cell treatment in MM.

Mass spectrometry vs immunofixation for treatment monitoring in multiple myeloma.

Monitoring of the monoclonal protein (M-protein) by electrophoresis and/or immunofixation (IFE) has long been used to assess treatment response in multiple myeloma (MM). However, with the use of highly effective therapies, the M-protein becomes frequently undetectable, and more sensitive methods had to be explored. We applied IFE and mass spectrometry (EXENT&FLC-MS) in serum samples from newly diagnosed MM patients enrolled in the PETHEMA/GEM2012MENOS65 obtained at baseline (n = 223), and after induction (n = 183), autologous stem cell transplantation (n = 173), and consolidation (n = 173). At baseline, the isotypes identified with both methods fully matched in 82.1% of samples; in the rest but 2 cases, EXENT&FLC-MS provided additional information to IFE with regards to the M-protein(s). Overall, the results of EXENT&FLC-MS and IFE were concordant in >80% of cases, being most discordances due to EXENT&FLC-MS+ but IFE- cases. After consolidation, IFE was not able to discriminate 2 cohorts with different median progression-free survival (PFS), but EXENT&FLC-MS did so; furthermore, among IFE- patients, EXENT&FLC-MS identified 2 groups with significantly different median PFS (P = .0008). In conclusion, compared with IFE, EXENT&FLC-MS is more sensitive to detect the M-protein of patients with MM, both at baseline and during treatment, and provides a more accurate prediction of patients' outcome. This trial was registered at www.clinicaltrials.gov as #NCT01916252.

High prevalence of diabetic retinopathy and lack of association with integrin α2 gene polymorphisms in patients with type 2 diabetes from Northeastern Mexico.

Diabetic retinopathy (DR) is one of the primary causes of blindness in the working age population and is characterized by angiogenesis in the retina. Platelets have been suggested to be involved in the pathogenesis of diabetic microvascular complications. The integrin receptor for collagen/laminin, α2ß1, mediates platelet primary adhesion to subendothelial tissues, which is an essential first step in thrombus formation. The gene encoding the α2 subunit of α2ß1 integrin has ≥8 polymorphisms, including a BglII/NdeI restriction fragment length polymorphism. To explore the prevalence of DR in a population from Northeastern Mexico, unrelated, hospitalized patients who had received a diagnosis of type 2 diabetes mellitus (DM2) at least 10 years previously were recruited (n=177). DR was diagnosed in a masked manner by independent ophthalmologists using fundus images captured using a non-mydriatic retinal camera. A total of 121 patients with DM2 (68%) had some degree of DR development (DR patients), and 56 patients with DM2 (32%) did not exhibit any sign of DR (No-DR patients). The results showed that after 15 years of DM2 progression, there is an increased risk of DR (P=0.0497; odds ratio, 1.993). In addition, insulin therapy and family history of DM2 were significantly associated with DR. In order to detect a possible association between DR and BglII/NdeI α2 gene polymorphisms, a comparative cross-sectional study between DR and No-DR patients was conducted. The α2 gene was genotyped by polymerase chain reaction-restriction fragment length polymorphism assay. Statistical analysis revealed no association between BglII/NdeI genotypes and the development of DR in this group of patients. In conclusion, the present data indicate a high prevalence of DR in the Mexican population and suggest that the damage in DR is due to other factors, such as the duration of the DM2, and is not linked to BglII/NdeI α2 gene polymorphisms.

Implementation of SintromacWeb(R), a new internet-based tool for oral anticoagulation therapy telecontrol: Study on system consistency and patient satisfaction.

Most computer- or internet-assisted systems for oral anticoagulation therapy (OAT) telemanagement have limitations when it comes to implementation within a healthcare center. It was the objective of this study to evaluate convenience and patient satisfaction with the use of SintromacWeb, a new OAT telecontrol system, compared with the conventional control. SintromacWeb consists of a point-of-care device for patient international normalized ratio (INR) self-testing and software that allows internet mediated interaction with physicians. Patients initiated the use of SintromacWeb and were followed up during a three-month period. A score-based questionnaire was completed in three controlled visits, and data were subsequently analysed. A total of 102 patients were enrolled. At first visit, 55.7% of the patients had their INR within normal range, and 64.9% at the final visit. Internal consistency of the questionnaire was good (Cronbach's a: 0.79). Scores in the questionnaire were independent of patient's age, education level, working status and INR value. The most valued features of SintromacWeb were: fewer visits to the hospital, simplicity and convenience of the system, and time administration for control tasks (86.7%, 82.7% and 77.6% of very satisfied patients, respectively). Also, patients showed indifference or were dissatisfied with the conventional system. At the final visit, 99% of patients declared that they were satisfied with their OAT control. Moreover, all patients continued using SintromacWeb after completion of the study. In conclusion, SintromacWeb telecontrol is a new model for management of anticoagulated patients. It was highly accepted and can be used by all patients regardless of their sociodemographic characteristics.

Validation of a procedure for the gas chromatography-mass spectrometry analysis of cocaine and metabolites in pericardial fluid.

A method is described for the simultaneous quantification of cocaine, benzoylecgonine, and cocaethylene in pericardial fluid. Pericardial fluid samples from autopsy casework involving cocaine-related deaths and deaths unrelated to drug abuse were collected. The extraction of cocaine and its metabolites was performed using Bond-Elut Certify columns. Pericardial fluid samples were adjusted to pH 7 and applied to the pre-conditioned cartridges. After the washing steps, compounds were eluted with a mixture of chloroform/isopropanol (80:20) with 2% ammonium hydroxide. The dry extracts were derivatized with pentafluoropropionic anhydride and hexafluoroisopropanol and analyzed by gas chromatography-mass spectrometry using electron impact ionization and selective ion monitoring acquisition. Deuterated internal standards were used. The analytical method developed was linear, sensitive, selective, accurate, and sufficiently precise to be applied routinely in forensic toxicology. In this study, the procedure has been successfully applied to a number of forensic cases involving cocaine intoxication.

Application of pericardial fluid to the analysis of morphine (heroin) and cocaine in forensic toxicology.

In this study opiates (morphine and codeine) and cocaine and its related metabolites (benzoylecgonine and cocaethylene) were analyzed in pericardial fluid by GC/MS. This is the first study reporting levels of drugs of abuse in this body fluid. The analytical method used has been previously validated and then applied to 54 drug-related deaths in the Barcelona area (Spain). Median levels were as follows: morphine 589ng/ml, range 19-8857 (n=49); codeine 26ng/ml, range 15-343 (n=35); cocaine 78ng/ml, range 10-220 (n=14), benzoylecgonine 742ng/ml, range 20-3386 (n=15), and cocaethylene 36ng/ml, range 9-100 (n=13). In addition, a comparative study of the concentration of opiates and cocaine in pericardial fluid by both semi-quantitative EMIT d.a.u. and GC/MS (used as reference) was performed. Fairly good correlations for opiates (r=0.905) and cocaine (r=0.859) were found; however, the consistently low results of EMIT in the analysis of cocaine comparing to GC/MS could be caused by matrix effect. In spite of that, it raises the possibility of using the immunoassay as a preliminary technique in forensic toxicology.

Erythrocyte deformability in survivors of acute myocardial infarction measured by two different methodologies.

In order to ascertain whether erythrocyte deformability (ED) is involved in chronic coronary syndromes, this rheological property was determined in 92 survivors of acute myocardial infarction (AMI) who had had the acute event 3 years ago and in 150 volunteers. From the 92 AMI survivors in 50 (43 males, 7 females aged 61+/-9 years) ED was determined with filtrometric techniques (Hanss Hemorheometre) and in 42 (32 males, 10 females aged 63+/-11 years) with laser diffractometric ones (Rheodyn SSD). The control group consisted of 66 and 84 volunteers whose ED was measured with the above mentioned devices respectively. Patients and controls were matched for age, sex, total cholesterol and triglyceride levels. With the Hanss Hemorheometre, the Rigidity Index (RI) was higher in patients than in controls (9.4+/-1.2 vs 8.7+/-1.5; p=0.01) although after adjusting for confounding variables (MCV and leukocyte count) in a logistic regression analysis the RI was no longer statistically significant. With the Rheodyn SSD the Erythrocyte Elongation Index (EEI) determined at 12, 30 and 60 Pa, did not show statistically significant differences between cases and controls at any of the shear stresses tested. Our results suggest that AMI survivors who had had the ischemic event 3 years ago do not show decrease RBC deformability with either of the two methodologies used. Red blood cell deformability does not appear to contribute to impaired microcirculatory blood flow in chronic coronary syndromes.