ALBERT

Description: On August 24, 2016, at 01:36 UTC a M W 6.0 earthquake struck an extensive area of the Central Apennines (Italy). It was followed by a large aftershock (M W 5.3, August 24, 02:33 UTC) and about 20 earthquakes with magnitude greater than 4.0, located between the towns of Norcia and Amatrice. Due to the mainshock magnitude and the widespread damaging level of buildings in the epicentral area, the Emersito task force has been mobilized by the Istituto Nazionale di Geofisica e Vulcanologia (INGV). The aim of Emersito is to carry out and coordinate the monitoring of local site effects, caused by geological and geomorphological settings. During the first days of the seismic emergency, Emersito installed a temporary seismic network for site effect studies at 4 municipalities close to the epicentral area (Amandola, Civitella del Tronto, Montereale and Capitignano), using 22 stations equipped with both velocimetric and accelerometric sensors. The selection of the sites where stations have been installed was mainly driven by the proximity to the epicentral area (without interfere with the rescue operation) and by peculiar geologic and geomorphologic settings (topographic irregularities, fault zones, alluvial plains).

Description: ThaDD regimen has provided significant results in recurrent/relapsed multiple myeloma (MM) patients (Offidani et al, 2006). In order to further improve those results without significantly increasing toxicity, we decided to include Velcade, synergic as per activity but not toxicity with the other drugs of ThaDD regimen. ThaDD-V was scheduled as follows: Thalidomide 100 mg/day, pegylated liposomal Doxorubicin 30 mg/sm iv days 4; Dexamethasone 20 mg days 1–2, 4–5, 8–9, 11–12 and Velcade 1.3 mg/sm iv days 1, 4, 8, 11 every 28 days (induction therapy). Patients received bortezomib for alternate cycles as following: 1 mg/sm day 1, 8, 15 and dexamethasone 20 mg days 1–2, 8–9, 15–16 and thalidomide 100 mg/day and dexamethasone 40 mg/day for 4 days monthly for a total of six cycles as consolidation therapy. Then patients received thalidomide 100 mg/day until relapse (maintenance therapy). Actually 20 patients (7 M, 13 F; median age 62.5 yrs, range 31–75) are assessable for response and toxicity. Five pts (25%) showed WHO performance status (PS) 〉 1, 9 pts (45%) presented refractory disease, 8 pts (40%) were priorily administered ≥ 2 lines of a treatment and 14 patients (70%) were submitted to one previous autologous stem cell procedure. Seven patients (35%) had extramedullary disease and 7 had unfavourable cytogenetics. Twelve patients scored an ISS ≥ 2 and 11 (55%) were in first remission for ≤ 12 months median duration. No patients were previously treated with Velcade, whereas six patients had received short-term Thalidomide treatment. Response was assessed according to IMVG uniform response criteria. Seventeen of 20 patients responded after at least one chemotherapy cycle reporting 5 (25%) sCR, 3 CR (15%), 8 VGPR (40%) and 1 stable disease. Three patients (15%) had extramedullary progressive disease. In a median follow-up of 12 months, 2 (10%) patients progressed and 1 (5%) died from cardiac infarction. Time to progression and overall survival were 73% and 95% at 12 months. We observed 4 grade 3 thrombocytopenia, 2 grade 3 DVT, 1 grade 3 diarrhoea, 1 grade 3 asthenia, 1 grade 4 infection and 1 grade 3 dermatological toxicity. Six patients developed grade 2 peripheral neuropathy and other three grade 3. In conclusion, ThaDD-V is extremely active in advanced MM patients as demonstrated by the elevated precentage of high quality remission. Nevertheless, patients are at substantial risk of developing neurotoxicity so the protocol was amended as per Velcade dose intensity and Thalidomide dose.

Description: Abstract 2437 Clinical course in individual CLL patients (pts) is highly heterogeneous. Several pts show an indolent disease and other experience an aggressive course rapidly succumbing to disease-related events. Different biomarkers have been developed to identify pts with evolving disease but their application is mostly limited to clinical trials. It has been shown that measurement of clonal serum free light chains (sFLC) levels, through a straightforward assay validated for plasma cell disorders, could independently identify CLL pts with progressive disease. By analyzing the largest cohort of CLL pts ever reported, we wished to: i) determine the independent predictive value of sFLC abnormalities in the context of established biomarkers; ii) define the predictive hierarchy of these markers; iii) incorporate sFLC into a novel prognostic system. Cryopreserved sera at diagnosis from 449 untreated pts (282 males and 167 females, median age 65 yrs; r 33–89) were collected for analysis at two reference laboratories. After quantization (Freelite, The Binding Site, Ltd., UK) of sFLC k and l levels (r, k: 3.3–19.4 mg/mL; l: 5.71–26.3 mg/mL), their ratio was calculated as sFLC k/l (r: 0.26–1.65). An abnormal sFLCk/l was found in 150/449 cases (132 k/18 l). A significant correlation emerged between sFLCk/l and CD38, ZAP70, unmutated IgHV and unfavourable FISH but not with Binet stage (Table). At a median follow up of 3 yrs (r 1–20), 149 of 449 pts were treated due to progressive disease (NCI Guidelines). Treatment-free survival (TFS) was significantly shorter (p

Description: Identification of patients at risk of early disease progression is the mainstay of tailored management in chronic lymphocytic leukemia (CLL). Although application of established biomarkers is limited by intrinsic detection/readout complexities, abnormality of κ and λ serum-free light chain ratio [sFLC (κ/λ)] was proposed as a straightforward prognosticator in CLL. By analyzing 449 therapy-naive patients, we show that an abnormal sFLC(κ/λ), along with CD38, ZAP-70, IGHV mutations, cytogenetics and stage, independently predicts treatment-free survival (TFS) but becomes prognostically irrelevant if the cumulative amount of clonal and nonclonal FLCs [sFLC(κ + λ)], a variable associated with cytogenetic risk, exceeds the threshold of 60.6 mg/mL. Patients with sFLC(κ + λ) above cut-off displayed a poorer TFS outcome, irrespective of sFLC(κ/λ). Only ZAP-70, cytogenetics, stage, and TFS remained associated with sFLC(κ + λ) in a multivariate model. By assigning 1 point each for these variables, the 3-year probability of TFS was 94.8%, 84.5%, 61.6%, and 21.1% for patients scoring 0, 1, 2, and 3 + 4, respectively (P 〈 .0001). These data, and the demonstration that monoclonal and polyclonal B cells concur to FLC synthesis in tumor tissues, suggest that sFLC(κ/λ) and sFLC(κ + λ) mirror distinct biologic processes in CLL. sFLC(κ + λ) assessment represents a sensitive and cost-effective tool for identifying CLL patients requiring early treatment.

Description: Abstract 267 Introduction: The tumor cells of HL, the Hodgkin and Reed-Sternberg (H-RS) cells, derive from germinal center B-cells with a deranged B-cell transcription program due to epigenetic silencing and acquired genetic lesions. Tissue H-RS cells are surrounded by a preponderant infiltrate of mixed non-malignant reactive cells, including B-lymphocytes, which provide essential signals for their survival and proliferation. Small percentages of B-cells, clonally related to H-RS cells, were found in the blood of HL patients (pts), suggesting they may represent putative HL-initiating cells (Jones, 2009). Since the serum free light chain (sFLC) assay detects and quantifies monoclonal and polyclonal B-cell populations expanding in lymphohemopoietic tissues, we exploited the sFLC testing to further explore the biologic significance of B-lymphocytes in HL. Patients and Methods: Frozen (-80°C) serum samples from 119 untreated cHL pts (48% males), with normal renal function and serum immunochemistry, were assayed by immunonephelometry (Freelite, The Binding Site, Ltd., UK). After quantization of free κ and λ concentrations (normal ranges, κ: 3.3-19.4 mg/L; λ: 5.71-26.3 mg/L), sFLC κ/λratio was calculated (reference range 0.26-1.65). The median age was 31 years (r, 15–70), with 22% aged ≥ 45 years. Histology was nodular sclerosis in 85 pts (71.4%) and 72 (60.5%) were in stage I–II. According to GHSG criteria, 16% had early favorable, 29% early unfavorable and 55% advanced disease. The International Prognostic Score (IPS) was of 0-2 and ≥ 3 in 66.4% and 33.6% of pts, respectively. Following ABVD (4-6 courses ± RT), the median Event-free survival (EFS) was 78 mo.s [95% CI, 68-88] for the entire population and 75 mo.s [95% CI, 60-90] and 64 mo.s [95% CI, 50-77] for pts in early and advanced stage, respectively. Results: Elevated κor λsFLC concentrations were found in 47% (median 29.55 mg/L; r, 19.44 - 64.70) and 29.4% (median 32.70 mg/L; r, 26.60 - 79.77) of pts, respectively. In 30 pts (25.2%) levels of polyclonal κ and λ sFLC were concurrently elevated. The sFLC κ/λratio was abnormal in only 7.5% of pts (clonal λ: 8/119; clonal λ: 1/119). The presence of high sFLC levels correlated with lymphopenia (〈 0.6 × 109/L; p= 0.04), leukocytosis (WBC 〉 15 × 109/L; p=0.03), ESR (〉 50; p=0.04) and unfavorable IPS (≥ 3; p=0.03), but not with EBV status and risk factors such as stage, B symptoms, bulky and extra nodal disease, LDH and albumin. The association with leukocytosis may in part result from inhibition of spontaneous neutrophils apoptosis exerted by FLC (Cohen, 2003). Most interestingly, while we found no significant association with response rate, baseline elevation in sFLC predicted for EFS in 51 evaluable pts with early stage disease. Patients were divided into tertiles and best break point value for predicting EFS coincided with the upper limit of the highest tertile for both κ (〉25.53 mg/L) or λsFLC (〉26.66 mg/L) (Figure 1). The pts in the top tertile had the worst outcome compared with the 2 lower tertiles (κ, p=0.015; λ, p=0.002). Outcomes for the 2 lower tertiles were comparable. Data remained significant after stratification for the IPS. In contrast, baseline sFLC levels, κ or λ, were not predictive for EFS in pts with advanced disease. Interestingly, sFLC levels in a control group of 30 pts in continuous CR from 2 years, were within the normal ranges. Conclusions: We have shown that about 50% of cHL pts displays elevated levels of sFLC mirroring the presence of a consistent polyclonal B-cell expansion at diagnosis. The role of reactive B-cells in HL is poorly understood. While some data indicate that high intratumoral B-cell counts may predict for a better outcome, the activity of rituximab in cHL, regardless of CD20 expression on H-RS cells, was suggested to result by depletion from the HL microenvironment of normal B lymphocytes required for tumor cell growth (Younes, 2003). Our study support that elevated sFLC levels may reflect an increased polyclonal B cell activity in cHL microenvironment which appears to negatively influence the outcome of pts in early stage disease. This effect is lost in advanced disease, suggesting that rituximab might result more active for pts in early than advanced stages. That putative HL-initiating small B-cells may emerge from the expanded polyclonal B-cell population present from the early phases of disease development, is an intriguing possibility. Disclosures: Marchei: Radim, Italy: Employment. Amoroso:The Binding Site, Ltd: Consultancy.

Description: The serum free light chain (sFLC) assay represents a predictor of prognosis and therapeutic response in monoclonal gammopathies and multiple myeloma (MM). Since sFLC testing detects and quantifies monoclonal and polyclonal B-cell populations expanding in lymphohemopoietic tissues, we evaluated the potential value of sFLC abnormalities as a novel disease marker for patients (pts) with B-cell non-Hodgkin’s lymphomas (NHL). Frozen (−80°C) serum samples from 354 untreated pts with NHL categorized according to WHO criteria (Table 1) and with normal renal function and serum immunochemistry, were assayed by immunonephelometry (Freelite, The Binding Site, Ltd., Birmingham, UK). After quantitation of serum free k and l concentrations (normal ranges, k: 3.3–19.4 mg/mL; l: 5.71–26.3 mg/mL), FLC ratio was calculated as k/l (free k concentration divided by free l, reference range 0.26–1.65). In cases with a ratio 〉1.65, k was considered the ‘involved’ (inv.) FLC and l the ‘uninvolved’ FLC, and vice versa if the ratio was less than 0.26. Sera from 45 MM pts, 43 pts with solid tumors and 22 pts with reactive lymphadenopathy (LAD) were also analyzed. Elevated sFLC concentrations, i.e. both k and/or l, were found in 53% to 80% of cases while abnormal FLC (k/l) ratios were detected in a restricted fraction of pts with a histotype-related fashion (Table). Tumors mostly arising from pre-germinal centre (GC) B-cells, i.e. Small Lymphocytic Lymphoma (SLL) and Mantle Cell Lymphoma (MCL), and those deriving from late post-GC cells, i.e. Burkitt lymphoma (BL), displayed the higher rates of abnormal FLC ratio (57%, 42% and 33% for MCL, SLL and BL, respectively), due to free k chain involvement in 86% to 〉90% of cases. Accordingly, these tumors showed a high median inv. k chain concentration (33.5 to 53 mg/L). In Follicular Cell (FCL) and Diffuse Large B Cell (DLBC) lymphomas the FCL ratio was abnormal in 23% and 25% of pts respectively, due to k FLC involvement in 〉 90% of cases, and a median serum inv. k concentration of 26 mg/mL in FCL (G1 20.5; G2, 16.3; G3, 16.9; G3b, 23,6 mg/L) and 25 mg/L in DLBCL. Interestingly, 8 cases of localized DLBCL of bone and CNS showed a normal sFLC test. Pts with disseminated Marginal Zone (MZ) lymphoma displayed abnormal FLC ratio in 16% of cases and the highest median inv. k concentration (66.5 mg/L). sFLC testing was positive 〉 80% of MM pts while no solid tumors and reactive LADs cases displayed abnormal sFLC ratio. In15 NHL pts (FCL, MCL, MZL) given upfront treatment with Rituximab (R) or Zevalin, sFLC test was explored as a tool for therapeutic monitoring. A stepwise decrease in inv. k chains levels was observed in all responders, followed by a rise at relapse. Similarly, disease control in SLL (n=14) and MCL (n=10) pts by first line R-immunochemotherapy was associated to normalization of sFCL test. Our results, on the largest serie reported, indicate that NHL pts display a high frequency of monoclonal sFLC. Differences among WHO histotypes at presentation may mirror specific biologic features, including pre/post-GC derivation and propensity to dissemination. sFCL testing represents a furher tool to dissect biologic heterogeneity of NHL and a new marker for therapeutic monitoring. SLL (n= 33) MCL (n=28) MZL (n=37) FCL (n=105) DLBCL (n=123) BL (n=20) * median concentration (mg/L) sFLC+ (%) 20 (60.6) 23 (82) 28 (75.6) 56 (53.3) 71 (57.7) 13 (65) Abnormal k/l (%) 14 (42.4) 16 (57) 6 (16) 24 (23) 31 (25.2) 6 (33) Involved k (%) 12 (85.7) 14 (88) 5 (83.3) 23 (95.8) 29 (93.5) 4 (66.6) Involved l (%) 2 (14.2) 2 (12.5) 1 (16.6) 1 (4.2) 2 (6.45) 2 (33) Involved k* (range) 53.0 (7.2–83.5) 33.5 (10.6–168.4) 65.5 (25.6–269.6) 25.96 (9.3–108.3) 25.2 (6.8–207.6) 47.6 (12.3–117.2) Involved l* (range) 133.7 (74.6–192.8) 75.3 (39.7–111) 318.28 215.4 97.5 (95–100) 135.4 (127.2–143.5)