ALBERT

Description: Abstract 5007 Introduction Non Hodgkin Lymphoma represent a category of hematological malignances which are chemo and radio-sensitive; improvements in their treatment had been achieved by immunotherapeutic approaches. However some patients will relapse after achieving complete remission (CR). Obviously, in order to detect and possibly treat them as soon as possible, a follow up strategy has to be planned. The more diffuse follow up have been planned years before the introduction of innovative methods and imaging techniques, suggesting the opportunity to revise these programs. In particulary it is not still clear which is best techniques useful to properly follow this patient. Recently new interesting methods are available like PET, CT-PET and minimal residual disease (MRD) monitoring. Methods 418 NHL patients -both low and high grade- treated at our institution from 1995 to 2005 who achieved a CR status according to Cheson criteria have been evaluated. LH NHL included follicular lymphoma, lymphoplasmocytic lymphoma, Marginal zone lymphoma, and small lymphocytes lymphoma. In the HG NHL, we included T-cells lymphomas, diffuse large cell lymphoma, lymphoblastic lymphoma, Mantle cell lymphoma, anaplastic lymphoma, Burkitt lymphoma. Patient characteristics are summarized in Table 1. Follow up is planned for 5 years divided in two periods: in the first two years patients are evaluated every 3 months and in the following three years every sixth month. At each visit physical examinations, blood testing (blood count, chemistry) are performed; for imaging techniques we alternate a whole body CT scans to ultrasounds and chest X-ray coupled. Bone marrow samples for both pathological and molecular analysis are collected every six months in the first period and once a year afterwards. PETs were usually performed when CT showed uncertain findings. Results There were 431 events, with 188 first relapses, 86 second, 18 third, 4 fourth and 1 fifth relapses. Relapse rate was similar among high and low grades, (37% and 35 % respectively) but time to relapse was longer for low grades (18.2 months vs 8.9 months). There was not relationship between IPI status and relapse rate. 72 % of relapse was at the same site of diagnosis. Relapses were detected by ultrasound in 139 cases (32 %), CT scans in 110 (25.5%) and by physical examination in 62 (14.4%). Remaining patients' relapse were diagnosed with other techniques (lab test, gastroscopy, NRM) New techniques as MRD monitoring, PET or PET/CT were not available for many patients, anyway MRD monitoring was able to detect disease re-appearance in 2%, and we had a total of 28 cases (6,5%) of relapse diagnosis with PET, but we noted a total of 18,5 % of false positive. Discussion and conclusions Many papers from literature raised many questions about which is the best techniques to follow patients. Many authors showed how symptoms onset and clinical findings appeared to be the more important for relapse detection compared to imaging before and during CT era. Some works pointed out also that even when CT detected earlier a relapse that do not translate in a survival advantage. Recently much interest has been focused on PET, CT-PET and MRD. They two appeared to be very important as prognostic tolls but their role for follow up purpose is still debatable. On the basis of clinical data and of these consideration routine PET is not recommended during follow up. Unfortunately PETs and MRD monitoring were not available for the majority of our patients, diagnosed in the nineteen's. In conclusion in our experience we observed some usefulness of CT scans and ultrasounds but we must recall that the majority of literature is not consistent with our results. Considering our experience and data from literature probably imaging should be performed routinely at the end of therapy, and during follow up only on the basis of presentation and clinical suspicion. As a matter of fact NCCN reviewed its guidelines do not suggesting a wide use of routine imaging. Further investigation by clinical and randomized trials are certainly needed to better understand, in particular the role of PET-PET/CT for follow up purpose. Disclosures No relevant conflicts of interest to declare.

Description: Previously untreated mantle cell lymphoma (MCL) are consistently associated with poor prognosis when treated with CHOP-like regimens. Typically the CR rate is 20–30%, median FFS = 10–16 months and median OS = 3 years. In the attempt to improve outcome we used a high dose intensity regimen such as Hyper-CVAD (HCVAD) with autologous stem cell transplant (ASCT). Twenty patients entered the study but only 10 up to now are valuable. Patients were apheresed after 2nd course of HCVAD and if apheresis (LPH) were PCR positive (Bcl1+/JH+) a second set of LPH were performed after completion of 4th cycle. To perform an in vivo purging Rituximab 375 mg/m2 was added at day +1 and +9 after last dose of ARA-C; GCSF 10μg/kg was commenced on day +5 until LPH was ultimated. Rituximab maintenance (375 mg/m2 once weekly for 4 consecutive weeks) started 2 month post-ASCT and was repeated every 6 months. Ten patients have completed 4 HCVAD and 7 /10 underwent ASCT and were conditioned with BEAM. After 4 HCVAD 7/10 patients were in CR and 3/7 in PR. After ASCT 1 PR obtained a CR, 1 PD obtained a VGPR and 5 CR maintained CR. Only 4CR post ASCT have received Rituximab maintenance and maintain CR. Two patients (1 CR blastic variant and 1 PR) refused ASCT and after 4 HCVAD received Rituximab maintenance and both are in CCR. Overall with a median follow up of 28.6 months (range 12–51) median survival is not reached. At 4 years 77.8% of patients are alive and PFS is 87.9%. Patients were monitored for bone marrow-MRD by PCR. Eight out of 10 were PCR+ at diagnosis and 7/8 were PCR negative after 4 HCVAD. After ASCT one PCR+ converted to PCR- and 1 PCR+ patient after 4 HCVAD converted to PCR- with Rituximab maintenance (refused ASCT). Conclusions: high dose intensity regimen HCVAD + Rituximab as in vivo purging and for maintenance allowed to collect tumor free grafts in 70% of PCR+patients at diagnosis and to reach an ORR of 100%, 7/10 CR and PR 3/10 (included 1 blastic variant). PCR negativity was obtained in 7/8 patients. One patient from PR converted to CR after ASCT and one after Rituximab maintenance (without ASCT); thus we might speculate that both high doses (BEAM) and Rituximab do play a role to increase the probability to obtain a CR and PCR-. Survival and PFS of 77.8% and 87.9% respectively at 4 years are encouraging. Further follow up and a higher enrolment of patients are needed to better define the role of HCVAD + Rituximab and ASCT to increase CR,PCR- PFS and OS in MCL.

Description: Despite recent advances, allografting remains the only potential cure for myeloma. From July 1999 to June 2005, 100 newly diagnosed patients younger than 65 years were enrolled in a prospective multicenter study. First-line treatment included vincristin, adriamycin, and dexamethasone (VAD)–based induction chemotherapy, a cytoreductive autograft (melphalan 200 mg/m2) followed by a single dose of nonmyeloablative total body irradiation and allografting from an human leukocyte antigen (HLA)–identical sibling. Primary end points were the overall survival (OS) and event-free survival (EFS) from diagnosis. After a median follow-up of 5 years, OS was not reached, and EFS was 37 months. Incidences of acute and chronic graft-versus-host disease (GVHD) were 38% and 50%, respectively. Complete remission (CR) was achieved in 53% of patients. Profound cytoreduction (CR or very good partial remission) before allografting was associated with achievement of posttransplantation CR (hazard ratio [HR] 2.20, P = .03) and longer EFS (HR 0.33, P 〈 .01). Conversely, development of chronic GVHD was not correlated with CR or response duration. This tandem transplantation approach allows prolonged survival and long-term disease control in patients with reduced tumor burden at the time of allografting. We are currently investigating the role of “new drugs” in intensifying pretransplantation cytoreduction and posttransplantation graft-versus-myeloma effects to further improve clinical outcomes. (http://ClinicalTrials.gov; NCT-00702247.)

Description: Abstract 4742 Introduction Neutropenic enterocolitis (NEC) is a life threatening complication of chemotherapy in leukemic and solid tumor patients with an incidence ranging from 2.6% to 33%. It is a necrotizing inflammatory disease that most commonly involves the ileo-cecal region. The cecum is almost invariably affected likely due to its distensibility and limited blood supply. Macroscopically the involved bowel segments show oedematous and thickened walls, with varying degrees of ulceration and haemorrhage. Perforation occurs in 5%-10% of cases. Early diagnosis is crucial to start conservative medical treatment which appears the optimal strategy. Criteria for prompt surgical treatment have also been proposed (Shamberger RC et al, 1986) so that a careful clinical evaluation by both the physician and the surgeon is mandatory (Davila ML et al, 2004). NEC should be always suspected in neutropenic patients with abdominal pain, fever and diarrhea. Ultrasound (US) has been used to evaluate bowel-wall thickening (BWT). One study correlated BWT with clinical outcome: 60% of patients with BWT 〉 10 mm died from NEC as compared with 4.2% of those with BWT 〈 10 mm (Cartoni C. et al, 2001). Overall, despite aggressive treatment, mortality rate is up to 21-48% and patients may die within hours from the onset of acute symptoms. Because of the risk of early death, a swift diagnosis is imperative. We investigated if US could detect early signs of NEC and lead to prompt treatment. We analyzed two patients cohorts (A and B). In cohort A US was performed later in the course of the disease whereas, in cohort B US was immediately performed at the onset of a single symptom (diarrhea and/or abdominal pain whichever occurred first with/without fever). Underlying haematological diagnoses were Hodgkin Disease (N=10), acute leukemias (N=9), multiple myeloma (N=3) and non-Hodgkin lymphomas (N=10). Treatments consisted of standard chemotherapy (N=10), myeloablative allografting after busulfan/cytoxan (N=1) and cytoxan/total body irradiation (N=1); autografting after busulfan/cytoxan (N=1), BEAM (N=16) and melphalan (N=3). Results All 32 patients showed grade 4 neutropenia at the onset of symptoms. Thirty-one/32 (97%) complained of abdominal pain whereas diarrhea was present in 30/32 (94%). Positive stool (for Clostridium Difficilis and Escherichia Coli) and blood cultures (62% Gram negative and 38% Gram positive bacteria) were found in 4/32 (12.5%) and 8/32 (25%) respectively. Trans-abdominal real-time US scanning of the bowel was performed using a 3.5-5 MHz convex probe and a 7 MHz linear transducer. A portable sonographer (Esaote model My Lab 25) was used for bed-side US. Both trans-abdominal axial and transverse US scans were performed on the colon. US signs of NEC were defined as thickening or dilation of small and/or large intestine. The intestinal involved areas were ileum, 21%, last terminal ileum, 21%, cecum 7%, ascending colon, 16%, transverse colon, 16%, descending colon, 14% and jejunum, 4%. US revealed signs of NEC in 8/14 in cohort A and 18/18 in cohort B. In cohort B, early US detected signs of NEC in 7 patients with abdominal pain and diarrhea without fever. Complete response to prompt medical treatment occurred in 6. Three/7 patients developed fever 48 hours after the US-guided diagnosis of NEC. Two patients, at risk of wall rupture by US imaging (confirmed by CT scans), underwent successful colon surgery within 12 hours from diagnosis. Except for these 2, all patients received medical treatment (broad spectrum antibiotics for gram-negative, gram-positive and anaerobic coverage, and antifungal drugs, granulocyte transfusions, bowel rest and granulocyte colony stimulating factor as per internal protocol). Overall 3/32 patients died. Conclusions Early bed-side intestinal US in neutropenic patients performed at the onset of a single symptom suggestive of NEC led to timely and successful treatment of this life threatening complication even before the development of fever. Disclosures: No relevant conflicts of interest to declare.

Description: A mini review of the toxicity of Thallium (Tl) at low doses is herein presented. Thallium has severe toxicity. Although its acute biological effects have been widely investigated and are well known, its biological effects on human health and in cell cultures at low doses (

Description: Metabolomic profiling of cell lines has shown many potential applications and advantages compared to animal models and human subjects, and an accurate cellular metabolite analysis is critical to understanding both the intracellular and extracellular environments in cell culture. This study provides a fast protocol to investigate in vitro metabolites of immortalized hippocampal neurons HN9.10e with minimal perturbation of the cell system using a targeted approach. HN9.10e neurons represent a reliable model of one of the most vulnerable regions of the central nervous system. Here, the assessment of their extracellular metabolic profile was performed by studying the cell culture medium before and after cell growth under standard conditions. The targeted analysis was performed by a direct, easy, high-throughput reversed-phase liquid chromatography with diode array detector (RP-HPLC-DAD) method and by headspace solid-phase microextraction–gas chromatography–mass spectrometry (HS-SPME-GC-MS) for the study of volatile organic compounds (VOCs). The analysis of six different batches of cells has allowed to investigate the metabolic reproducibility of neuronal cells and to describe the metabolic “starting” conditions that are mandatory for a well-grounded interpretation of the results of any following cellular treatment. An accurate study of the metabolic profile of the HN9.10e cell line has never been performed before, and it could represent a quality parameter before any other targeting assay or further exploration.