Rapid determination of anti-estrogens by gas chromatography/mass spectrometry in urine: Method validation and application to real samples

AbstractA fast screening protocol was developed for the simultaneous determination of nine anti-estrogenic agents (aminoglutethimide, anastrozole, clomiphene, drostanolone, formestane, letrozole, mesterolone, tamoxifen, testolactone) plus five of their metabolites in human urine. After an enzymatic hydrolysis, these compounds can be extracted simultaneously from urine with a simple liquid–liquid extraction at alkaline conditions. The analytes were subsequently analyzed by fast-gas chromatography/mass spectrometry (fast-GC/MS) after derivatization. The use of a short column, high-flow carrier gas velocity and fast temperature ramping produced an efficient separation of all analytes in about 4min, allowing a processing rate of 10samples/h. The present analytical method was validated according to UNI EN ISO/IEC 17025 guidelines for qualitative methods. The range of investigated parameters included the limit of detection, selectivity, linearity, repeatability, robustness and extraction efficiency. High MS-sampling rate, using a benchtop quadrupole mass analyzer, resulted in accurate peak shape definition under both scan and selected ion monitoring modes, and high sensitivity in the latter mode. Therefore, the performances of the method are comparable to the ones obtainable from traditional GC/MS analysis. The method was successfully tested on real samples arising from clinical treatments of hospitalized patients and could profitably be used for clinical studies on anti-estrogenic drug administration

Long-term trends of PM10-bound arsenic, cadmium, nickel, and lead across the Veneto region (NE Italy)

Since the mid-90s, the European Community has adopted increasingly stringent air quality standards. Consequently, air quality has generally improved across Europe. However, current EU standards are still breached in some European hotspots. The Veneto region (NE Italy) lies in the eastern part of the Po Valley, a major European hotspot for air pollution, where EU standards for particulate matter, nitrogen oxides and ozone are still breached at some sites. This study aims to analyse the PM10-bound arsenic, cadmium, nickel, and lead concentrations over a 10 years-long period (2010-2020) in the Veneto Region by using data collected by the local environmental protection agency (ARPAV) in 20 sampling stations mostly distributed across the plain areas of the region and categorized as rural (RUR), urban (URB), and suburban (SUB) background, industrial (IND) and traffic (TRA) hotspots (Figure 1). The comprehensive dataset discussed in this study was statistically investigated to detect the seasonal trends, their relationship with other air pollutants and meteorological parameters and their spatial variations at a regional scale. This study completes previous air quality studies over the Veneto region for gaseous pollutants and bulk PM10 (Masiol et al. 2017). Samplings were carried out according to CEN EN 12341:1998 standard on quartz fibre filters and were continuous for 24 h, starting at midnight. The gravimetric determination of PM10 mass was measured following the CEN EN 12341:2014 standard. The elemental analysis was performed using an ICP-MS (Agilent 7700) after acid digestion (EN 14902:2005). The trends were analysed using different approaches on the monthly-averaged data. The shape of trends and their seasonal variations were assessed through the seasonal-trend decomposition time series procedure based on “Loess” (STL). The linear trends were computed by the Mann-Kendall trend test (p < 0.05) and the Theil-Sen nonparametric estimator of slope (MK-TS). Since this latter analysis assumes monotonic linear trends and does not consider the shape of trends, the presence of possible breakpoints was investigated using the piecewise regression. Generally, monthly patterns of all analysed elements show higher concentrations during winter, following PM10 concentrations. Some exceptions were detected and discussed. Results of trend analysis indicate statistically significant negative (decreasing) or null linear trends in almost all stations. A few positive (increasing) but not statistically significant trends were also detected. Some sites showed rapid decreases occurred in short periods and linked to peculiar events or local causes. Among others, several sites across the Venice area showed significant drops of arsenic concentrations after the REACH (Registration Evaluation Authorisation of Chemicals) implementation (Formenton et al., 2021). Data used in this study are provided by ARPAV (Agenzia Regionale per la Prevenzione e Protezione Ambientale del Veneto, https://www.arpa.veneto.it/)

551 Pegasus Skin, a study of SAR444245 (THOR-707, a pegylated recombinant non-alpha IL2) with cemiplimab for the treatment of participants with advanced unresectable or metastatic skin cancers

BackgroundSAR444245 (THOR-707) is a recombinant human IL-2 molecule that includes a PEG moiety irreversibly bound to a novel amino acid via click chemistry to block the alpha-binding domain while retaining near-native affinity for the beta/gamma subunits. In animal models, SAR444245 showed anti-tumor benefits, but with no severe side effects, both as single agent and when combined with anti-PD1 comparing with historical data from aldesleukin. The HAMMER trial, which is the FIH study, shows preliminary encouraging clinical results: initial efficacy and safety profile with SAR444245 monotherapy and in combination with pembrolizumab supporting non-alpha preferential activity, validating preclinical models. The Pegasus Skin Phase 1/2 study will evaluate the clinical benefit of SAR444245 in combination with cemiplimab (anti-PD1) for the treatment of participants with cutaneous squamous cell carcinoma (CSCC) or melanomaMethodsPegasus Skin (NCT04913220) will enroll approximately 80 participants in 2 separate cohorts. In cohort A, participants with a locally advanced, unresectable or metastatic melanoma will receive SAR444245 + cemiplimab as first line (1L) therapy. In cohort B, participants with locally advanced or metastatic CSCC who have not received moe than 2 prior lines of systemic therapy and are not candidates for curative surgery or radiation will receive SAR444245 + cemiplimab. The study will start with a dose escalation to determine the recommended phase 2 dose (RP2D) of SAR444245 when combined with cemiplimab. The starting dose will be 16 μg/kg Q3W (DL1) with a possibility to de-escalate to 8 μg/kg Q3W (DL -1) or escalate to 24 μg/kg Q3W (DL2) based on the occurrence of DLT and overall assessment of safety. Participants enrolled in the Dose Escalation and treated at the RP2D selected for Dose Expansion will be included in the total number of participants for efficacy and safety evaluation. Participants will receive study treatment until disease progression, unacceptable toxicity, or completion of 35 cycles. Cemiplimab will be administered 350 mg per label, Q3WAcknowledgementsThe Pegasus Skin study is sponsored by SanofiTrial RegistrationNCT04913220Ethics ApprovalAll applicable ECs are obtainedConsentAll participant consents are obtaine

Drug-Related Cardiotoxicity for the Treatment of Haematological Malignancies in Elderly.

Meyer Paul Hugo. Diderot oder die Ambivalenz der Aufklärung, Neumann, 1987. In: Recherches sur Diderot et sur l'Encyclopédie, n°4, 1988. pp. 164-165

Proximal Gastrojejunal Reconstruction after Pancreaticoduodenal Resection

Introduction. Reconstruction by proximal gastrojejunostomy, and distal biliary and pancreatic anastomoses is infrequently used after resection of the head of the pancreas because of fear of fistulas and cholangitis. Pancreaticoduodenectomy is being performed more frequently for cystic malignant and premalignant lesions. Because of this there is a need for endoscopic visualization and biopsy of the residual pancreatic duct, since multi-centricity is characteristic of some of these malignancies. Since endoscopic access of the bile duct and pancreatic duct is difficult and unsuccessful in 50–70% after B II or Roux Y reconstruction, we prospectively studied the merit and complications (early and late) of proximal gastrojejunal (PGJ) reconstruction after pancreaticoduodenal resection. Material and Methods. Thirty nine consecutive, non-radomized patients underwent pancreaticoduodenectomy and PGJ reconstruction over 14 mos. There were 21 males and 18 females. Results. 7 patients with IPMN have undergone repeat CT scanning for surveillance, with 3 requiring repeat EUS and ERCP. There were no technical difficulties accessing the pancreas or the pancreatic duct, supporting the PGJ reconstruction. Conclusion. Proximal gastrojejunal reconstruction following pancreaticoduodenal resection may be safely done with similar morbidity to traditional pancreaticojejunal reconstructions. PGJ reconstruction may be of greater value when direct visual access to the bile duct or pancreatic duct is necessary, and should be considered when doing resection for mucinous cysts or IPMN of the head of the pancreas

Chondrogenic potential of chondrocytes in hyaluronic acid/PEG-based hydrogels is dependent on the hyaluronic acid concentration

Purpose: Hydrogels based on PEG and methacrylated poly(N-(2-hydroxypropyl) methacrylamide-mono/dilactate) (M10P10) are promising biomaterials for Biofabrication of cartilage constructs. Addition of hyaluronic acid (HA) to a hydrogel improves printability by increasing the viscosity. Methacrylating HA (HAMA) can ensure covalent binding in M10P10 hydrogels after UV-cross-linking. Chondrocytes can interact with HAMA via their CD44 receptor, however, the influence of HAMA on chondrogenic potential is unclear. This study aimed to evaluate the influence of different HAMA concentrations on chondrogenesis of chondrocytes in M10P10/HAMA hydrogels. Materials & Methods: Equine chondrocytes were encapsulated in M10P10 hydrogels containing different HAMA concentrations. Cylindrical constructs were cast, UV-cross-linked, and cultured in TGF-β-containing medium. Constructs were analyzed for evidence of cartilage formation. Results: Preliminary data showed an increase in glycosaminoglycan (GAG)/DNA for constructs with low HAMA concentrations (0.1-0.25%) while no differences were found for higher HAMA concentrations, compared to hydrogels without HAMA (Figure 1a). Further, constructs without or with low HAMA concentrations (0.1-0.5%) demonstrated collagen type II positive areas, while this was less pronounced in constructs with 0.5-1% HAMA (n=3, Figure 1b). Conclusion: Preliminary results indicate a dose-dependent effect of HAMA on chondrogenesis of chondrocytes: low concentrations (0.1-0.25%) increase GAG production while higher concentrations (0.5-1%) have no effect on GAG production and reduce collagen type II synthesis. Ongoing evaluations will reveal the extent of chondrogenesis and its association with HAMA concentrations in M10P10/HAMA, and the mechanism responsible for the dose-dependent effect. This study will impact the use of HAMA as viscosity enhancer to improve the printability of hydrogel

Cd19 cell count at baseline predicts b cell repopulation at 6 and 12 months in multiple sclerosis patients treated with ocrelizumab

Background: The kinetics of B cell repopulation in MS patients treated with Ocrelizumab is highly variable, suggesting that a fixed dosage and time scheduling might be not optimal. We aimed to investigate whether B cell repopulation kinetics influences clinical and radiological outcomes and whether circulating immune asset at baseline affects B cell repopulation kinetics. Methods: 218 MS patients treated with Ocrelizumab were included. Every six months we collected data on clinical and magnetic resonance imaging (MRI) activity and lymphocyte subsets at baseline. According to B cell counts at six and twelve months, we identified two groups of patients, those with fast repopulation rate (FR) and those with slow repopulation rate (SR). Results: A significant reduction in clinical and radiological activity was found. One hundred fifty-five patients had complete data and received at least three treatment cycles (twelve-month follow-up). After six months, the FR patients were 41/155 (26.45%) and 10/41 (29.27%) remained non-depleted after twelve months. FR patients showed a significantly higher percentage of active MRI scan at twelve months (17.39% vs. 2.53%; p = 0,008). Furthermore, FR patients had a higher baseline B cell count compared to patients with an SR (p = 0.02 and p = 0.002, at the six-and twelve-month follow-ups, respectively). Conclusion: A considerable proportion of MS patients did not achieve a complete CD19 cell depletion and these patients had a higher baseline CD19 cell count. These findings, together with the higher MRI activity found in FR patients, suggest that the Ocrelizumab dosage could be tailored depending on CD19 cell counts at baseline in order to achieve complete disease control in all patients

Residual vein thrombosis for assessing duration of anticoagulation after unprovoked deep vein thrombosis of the lower limbs: the extended DACUS study.

Abstract The safest duration of anticoagulation after idiopathic deep vein thrombosis (DVT) is unknown. We conducted a prospective study to assess the optimal duration of vitamin K antagonist (VKA) therapy considering the risk of recurrence of thrombosis according to residual vein thrombosis (RVT). Patients with a first unprovoked DVT were evaluated for the presence of RVT after 3 months of VKA administration; those without RVT suspended VKA, while those with RVT continued oral anticoagulation for up to 2 years. Recurrent thrombosis and/or bleeding events were recorded during treatment (RVT group) and 1 year after VKA withdrawal (both groups). Among 409 patients evaluated for unprovoked DVT, 33.2% (136 of 409 patients) did not have RVT and VKA was stopped. The remaining 273 (66.8%) patients with RVT received anticoagulants for an additional 21 months; during this period of treatment, recurrent venous thromboembolism and major bleeding occurred in 4.7% and 1.1% of patients, respectively. After VKA suspension, the rates of recurrent thrombotic events were 1.4% and 10.4% in the no-RVT and RVT groups, respectively (relative risk = 7.4; 95% confidence interval = 4.9-9.9). These results indicate that in patients without RVT, a short period of treatment with a VKA is sufficient; in those with persistent RVT, treatment extended to 2 years substantially reduces, but does not eliminate, the risk of recurrent thrombosis

The Use of Social Media and Digital Devices Among Italian Neurologists

Background: Digital devices and online social networks are changing clinical practice. In this study, we explored attitudes, awareness, opinions, and experiences of neurologists toward social media and digital devices. Methods: Each member of the Italian Society of Neurology (SIN) participated in an online survey (January to May 2018) to collect information on their attitude toward digital health. Results: Four hundred and five neurologists participated in the study. At work, 95% of responders use the personal computer, 87% the smartphone, and 43.5% the tablet. These devices are used to obtain health information (91%), maintain contact with colleagues (71%), provide clinical information (59%), and receive updates (67%). Most participants (56%) use social media to communicate with patients, although 65% are against a friendship with them on social media. Most participants interact with patients on social media outside working hours (65.2%) and think that social media have improved (38.0%) or greatly improved (25.4%) the relationship with patients. Most responders (66.7%) have no wearable devices available in clinical practice. Conclusion: Italian neurologists have different practices and views regarding the doctor–patient relationship in social media. The availability of digital devices in daily practice is limited. The use of social networks and digital devices will increasingly permeate into everyday life, bringing a new dimension to health care. The danger is that advancement will not go hand in hand with a legal and cultural adaptation, thus creating ambiguity and risks for clinicians and patients. Neurologists will need to be able to face the opportunities and challenges of this new scenario