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Background: The combination of endocrine therapy with the cyclin-dependent kinases 4/6 inhibitor palbociclib was proven to be effective for the treatment of hormone receptor (HR)-positive/HER2-negative metastatic breast cancer (MBC). Although generally well tolerated, treatment delays and dose reductions are frequently observed with palbociclib, mainly due to severe neutropenia. Predictors of palbociclib-related toxicities are still unknown, as well as the clinical relevance of its relative dose intensity (RDI). Henceforth, this study aimed to identify baseline clinicopathological features associated with a RDI Methods: We reviewed data of 150 consecutive patients with HR-positive/HER2-negative MBC patients treated with palbociclib at two Italian cancer centers from 2017 to 2019. Eligible patients must have received at least 3 cycles of treatment. Those who experienced early suspension due to unacceptable toxicities were still eligible. RDI was calculated as the ratio of actual dose intensity (cumulative administered dose/treatment duration) and planned dose intensity (cumulative planned dose/planned treatment duration). The association of both RDI Results: Overall, 142 patients were deemed eligible. Of these patients, 98 (69.0%) were treated with palbociclib plus fulvestrant, 44 (31.0%) with palbociclib plus aromatase inhibitors, and 73 (51.4%) in the first-line setting. The median number of administered palbociclib cycles was 8 (range: 1-24) and 61 patients (43.0%) required at least a first-level dose reduction (29 within 3 months). Furthermore, the median time to first dose reduction was 3.22 months, with neutropenia being responsible for 85.24% of first-level dose reductions. In the whole cohort, median RDI was 90.5% (95.1% for patients without dose reduction and 80% for those who had received a dose reduction). Notably, 28 patients (19.7%) experienced a RDI Conclusions: In our analysis, baseline weight and WBC were statistically associated with a RDI Citation Format: Giacomo Pelizzari, Elisa Bertoli, Marco Giavarra, Carla Corvaja, Lorenzo Gerratana, Debora Basile, Michele Bartoletti, Camilla Lisanti, Lucia Bortot, Silvia Buriolla, Mattia Garutti, Debora Avoledo, Marta Bonotto, Lucia Da Ros, Silvia Bolzonello, Mauro Mansutti, Paola Di Nardo, Gianpiero Fasola, Simon Spazzapan, Alessandro M Minisini, Fabio Puglisi. Predictors of relative dose intensity and early dose reduction in patients with metastatic breast cancer treated with palbociclib and endocrine therapy [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P5-14-08.

Simple Summary Immunotherapy has revolutionized the therapeutic landscape of melanoma. However, the absence of clinically validated predictive biomarkers is one of the major causes of unpredictable efficacy of immunotherapy. Indeed, the availability of predictive biomarkers could allow a better stratification of patients, suggesting which type of drugs should be used in a certain clinical context and guiding clinicians in escalating or de-escalating therapy. However, the difficulty in obtaining clinically useful predictive biomarkers reflects the deep complexity of tumor biology. Herein, we review the available literature to depict the most useful or promising biological biomarker able to predict immunotherapy response in melanoma. We also make a meta-analysis regarding PDL1 expression in melanoma and immune checkpoint response. Abstract Immunotherapy has revolutionized the therapeutic landscape of melanoma. In particular, checkpoint inhibition has shown to increase long-term outcome, and, in some cases, it can be virtually curative. However, the absence of clinically validated predictive biomarkers is one of the major causes of unpredictable efficacy of immunotherapy. Indeed, the availability of predictive biomarkers could allow a better stratification of patients, suggesting which type of drugs should be used in a certain clinical context and guiding clinicians in escalating or de-escalating therapy. However, the difficulty in obtaining clinically useful predictive biomarkers reflects the deep complexity of tumor biology. Biomarkers can be classified as tumor-intrinsic biomarkers, microenvironment biomarkers, and systemic biomarkers. Herein we review the available literature to classify and describe predictive biomarkers for checkpoint inhibition in melanoma with the aim of helping clinicians in the decision-making process. We also performed a meta-analysis on the predictive value of PDL-1.

Background Endocrine therapy (ET) plus cyclin-dependent-kinases 4/6 inhibitors (CDK4/6i) represents the standard treatment for luminal-metastatic breast cancer (MBC). However, prospective head-to-head comparisons are still lacking for 1st line (L) options, and it is still crucial to define the best strategy between 1st and 2nd L. Materials and methods 717 consecutive luminal-MBC pts treated between 2008 and 2020 were analyzed at the Oncology Department of Aviano and Udine, Italy. Differences about survival outcomes (OS, PFS and PPS) were tested by log-rank test. The attrition rate (AR) between 1st and 2ndL was calculated. Results At 1stL, pts were treated with ET (49%), chemotherapy (CT) (31%) and ET-CDKi (20%) while, at 2ndL, 33% received ET, 33% CT and 8% ET-CDKi. Overall AR was 10%, 7% for CT, 8% for ET and 17% for ET-CDKi. By multivariate analysis, 1stL ET-CDK4/6i showed a better mPFS1 and OS. Moreover, 2ndL ET-CDK4/6i demonstrated better mPFS2 compared to ET and CT. Notably, 1stL ET-CDKi resulted in higher mPFS than 2ndL ET-CDKi. Intriguingly, 1stL ET-CDK4/6i was associated with worse mPPS compared to CT and ET. Secondarily, 1stL ET-CDK4/6i followed by CT had worse OS compared to 1stL ET-CDK4/6i followed by ET. Notably, none of baseline characteristics at 2ndL influenced 2ndL treatment choice (ET vs. CT) after ET-CDKi. Conclusion Our real-world data demonstrated that ET-CDKi represents the best option for 1stL luminal-MBC compared to ET and CT. Also, the present study pointed out that 2ndL ET, potentially combined with other molecules, could be a feasible option after CDK4/6i failure, postponing CT on later lines., Highlights • To define the best treatment strategy for 1st and 2nd L is crucial for luminal mBC. • 1st L CDK4/6i-based therapies represent the standard treatment in luminal mBC. • This study confirmed the key role of 1st L CDK4/6i on a real-world cohort. • This study suggested a preeminent role for ET after first-line CDK4/6i. • The 2nd line ET with other molecules could allow to further postpone CT.

Background Aging induces meaningful changes in the immune system and inflammation response with increase in monocyte-lymphocyte ratio (MLR) and serum lactate dehydrogenase (LDH) levels. Aim of this study was to explore the prognostic role of MLR and LDH levels in older patients (pts) with metastatic colorectal cancer (mCRC). Methods We conducted a retrospective analysis of a consecutive cohort of 168 older (>70 years) patients with mCRC. The prognostic impact of MLR and LDH levels on overall survival (OS) was investigated through uni-and multivariate Cox regression analyses. Moreover, we categorized patients into three groups according to MLR and LDH levels (group 1: MLR-low and LDH-low; group 2: MLR-high or LDH-high; group 3: MLR-high and LDH-high). Results By univariate analysis, high LDH level (HR 1.74, 95% CI 1.05–2.90) and high MLR level (HR 2.19, 95% CI 1.48–3.44) were significantly associated with a worse OS. Conversely, primary tumor resection and left-sidedness were significantly associated with a longer OS. By multivariate analysis, high LDH level (HR 2.00, 95% CI 1.13–3.55) and high MLR level (HR 2.99, 95% CI 1.68–5.33) were independent prognostic factors of worse prognosis. Compared to group 1, a shorter survival was reported for patients included in group 2 (HR 1.97, 95% CI 1.21–3.23 in univariate; HR 2.54, 95% CI 1.43–4.51 in multivariate) or in group 3 (HR 2.42, 95% CI 24–4.74, p = .010 in univariate; HR 5.59, 95% CI 2.15-14.54 in multivariate) Conclusions High baseline levels of LDH, MLR or both are independent unfavorable prognostic factors in older patients treated with first-line chemotherapy for mCRC.

Different de-escalation strategies have been proposed to limit the risk of cumulative toxicity and guarantee quality of life during the treatment trajectory of patients with metastatic colorectal cancer (mCRC). Programmed treatment interruptions, defined as drug holidays (DHs), have been implemented in clinical practice. We evaluated the association between DHs and overall survival (OS). This was a retrospective study, conducted at the University Hospital of Udine and the IRCCS CRO of Aviano. We retrieved records of 608 consecutive patients treated for mCRC from 1 January 2005 to 15 March 2017 and evaluated the impact of different de-escalation strategies (maintenance, DHs, or both) on OS through uni- and multivariate Cox regression analyses. We also looked at attrition rates across treatment lines according to the chosen strategy. In our study, 19.24% of patients received maintenance therapy, 16.12% DHs, and 9.87% both, while 32.07% continued full-intensity first-line treatment up to progression or death. In uni- and multivariate analyses first-line continuous treatment and early discontinuation (treatment for less than 3 months) were associated to worse OS compared to non-continuous strategies (HR, 1.68, 95% CI, 1.22–2.32, p = 0.002 and HR,4.89, 95% CI, 3.33–7.19, p &lt, 0.001, respectively). Attrition rates were 22.8%, 20.61%, and 19.64% for maintenance, DHs, or both, respectively. For continuous therapy and for treatment of less than 3 months it was 21.57% and 49%. De-escalation strategies are safe and effective options. DHs after initial induction chemotherapy may be considered in clinically selected patients with metastatic colorectal cancer.

e13008 Background: Liquid biopsy-based biomarkers, including circulating tumor DNA (ctDNA), are increasingly important for the characterization of metastatic breast cancer (MBC). Currently, CA15.3 is the most commonly used serum marker for monitoring disease burden. To date, no liquid biopsy-based biomarkers have been proposed for this scope in clinical practice. Methods: The CRO-2018-56 multicenter study prospectively enrolled 83 patients (pts) with luminal-like MBC treated with first line endocrine therapy (ET) and CDK4/6 inhibitors. All pts were characterized for ctDNA through droplet digital PCR (ddPCR) in from 2018 to 2021. Clinicopathological and laboratory characteristics at baseline were tested for associations with tumor markers, ACTB fragments lengths, methylation status of ESR1 main promoters (expressed as promA and promB ratio, i.e., met_ratio) and ESR1/PIK3CA mutational status through Kruskal-Wallis test and Chi-square test. Prognosis was tested in terms of Progression Free Survival (PFS) and Overall Survival (OS) through log-rank test. Results: At baseline, in 26 (31%) pts disease was diagnosed as de novo metastatic, 66 (80%) pts had < 3 of metastatic sites, and 41 (49%) pts had < 5 of metastatic lesions. Bone metastases were detected in 53 (64%) pts, liver metastases in 21 (25%) pts, and lung lesions in 30 (36%) pts. A ctDNA-detected ESR1 mutation and a PIK3CA mutation were found in 15% of pts and in 34% pts, respectively. Met_ratio was > 1.5 in 35 (42%) of pts. Median CA15.3 was 48.2 U/mL and median CEA was 3.8 U/mL. Number of liver metastases and number of metastatic sites were significantly higher in pts with ESR1 mutation (respectively, P = 0.0055 and P = 0.0208). CA15.3 and ctDNA yield were significantly higher in pts with number of metastatic sites ≥ 3, (respectively, P = 0.0164, and P = 0.0239), while number of metastatic sites ≥ 3 and number of metastatic lesions ≥ 5 were significantly associated with CEA > 3.8 U/mL (respectively, P = 0.039, and P = 0.029). Presence of bone metastases was significantly associated with PIK3CA mutation (P = 0.040), while number of metastatic sites ≥ 3 was significantly associated with ESR1 mutation (P = 0.022). No association with tumor burden was observed for different ACTB DNA fragments lengths. Met_ratio > 1.5 was significantly associated with lower number of metastatic lesions (P = 0.031). Number of metastatic sites ≥ 3, high ctDNA yield and CEA were associated with worse OS (respectively P = 0.0465, P = 0.0250 and P = 0.0474), while only CEA impacted on PFS (P = 0.0097). Conclusions: In pts with luminal-like MBC, some liquid biopsy-based biomarkers (i.e., ctDNA-detected ESR1 and PIK3CA mutations, ctDNA yield) were significantly associated with the burden of disease. The potential clinical validity and utility of these results deserve to be tested in an expansion cohort.

e16102 Background: Lactate dehydrogenase (LDH) is recognized as indirect marker of tumor hypoxia and angiogenesis for several solid tumors. Ramucirumab is the first antiangiogenic agent approved for second-line therapy in advanced gastric cancer (GC), alone or in association with chemotherapy. To date, no reliable biomarkers can predict the potential benefit from anti-VEGFR2 treatment. This retrospective multicenter study aimed to assess the prognostic role of baseline LDH levels in patients (pts) undergoing second-line treatment for advanced GC. Methods: The study analyzed a retrospective cohort of consecutive advanced GC pts treated with second line therapy at IRCCS, CRO of Aviano and at Mauriziano Hospital of Torino, Italy, from 2010 to 2020. LDH levels prior to second-line treatment were classified as low-normal or high and standardized according to the upper limit of the reference range. To better determine the optimal LDH cut-off value, ROC analysis was performed (using PFS < 3 months as binary outcome). Normalized LDH values were subsequently sorted according to the ROC curve cut-off in order to test the association with overall survival (OS) through the Kaplan-Meier method and compared using the Log-Rank test. A multivariate Cox regression analysis assessed the prognostic impact of normalized LDH levels for OS calculated from the start of second line treatment. Results: Overall, 125 pts were enrolled. Of these, 81 pts (64.80%) received ramucirumab alone or plus paclitaxel as second line treatment, while 44 (35.20%) had taxanes or fluoropyrimidines combined with irinotecan. Median age was 68 years, 93% had an ECOG PS ≤ 1, 64.80% was first diagnosed with metastatic disease and 40.80% underwent primary tumor resection. Median second line PFS and OS were 4.2 and 7.9 months, respectively. Baseline LDH values were available for 99 pts. ROC analysis identified a normalized LDH value of 0.83 as the optimized cut-off point to define pts with poor prognosis. At univariate analyses, surgery of the primary tumor (HR 0.51, 95% CI 0.31-0.82, p = 0.005) was associated with better OS, while low-normal BMI ( < 25 kg/m2) at second line start (HR 1.99, 95% CI 1.21-3.28, p = 0.007), normalized LDH values ≥ 0.83 (HR 1.86, 95% CI 1.13-3.07, p = 0.015) and PS > 1 (HR 5.65, 95% CI 2.29-13.92, p < 0.001) were associated with poorer outcome. At multivariable model, only PS > 1 was independently associated with poor prognosis (HR 9.93, 95% CI 3.27-30.08, p < 0.001). Subgroup analyses showed no significant heterogeneity in OS outcome according to normalized LDH levels between patients treated with or without ramucirumab. Conclusions: Elevated LDH levels are indicators of worse outcome in advanced gastric cancer pts. Notably, the LDH cut-off value identified a poor prognosis subgroup among pts with normal baseline levels, suggesting that current laboratory ranges could be suboptimal for patient stratification.

Neuroblastoma rat sarcoma (NRAS) mutation, occurring in about 20%-30% of cutaneous melanomas, leads to activation of RAS-RAF-MAPK cascade and represents a clear distinct clinicopathological entity in melanoma. In contrast with BRAF mutant melanoma, no specific target therapies are available outside the setting of clinical trials. In the field of immunoncology, the predictive role of NRAS mutation with respect to checkpoint inhibitors treatment has not clearly established and deserves further investigation. At present, the standard treatment is the same as for BRAF wild type melanoma. Ongoing trials are exploring novel combination strategies among patients with advanced NRAS mutant melanoma.

The reach and grasp follow different developmental trajectories, but are often considered to have achieved nearly adult-like precision and integration by 12 months of age. This study used frame-by-frame video analysis to investigate whether increasing precision demands, by placing small reaching targets on a narrow pedestal rather than on a flat table, would influence the reach and grasp movements of 12-month-old infants in a complementary or differential fashion. The results reveal that placing the target atop a pedestal impaired the infants’s ability to direct an appropriate digit towards the small target, but did not produce a corresponding decrease in the frequency with which they used an index-thumb pincer grip to grasp the target. This was due to the fact that, although infants were more likely to contact the target with a suboptimal part of the hand in the pedestal condition, a greater proportion of these suboptimal contacts ultimately transitioned to a successful index-thumb pincer grip. Thus, increasing task precision demands impaired reach accuracy, but facilitated index-thumb grip formation, in 12-month-old infants. The differential response of the reach and grasp to the increased precision demands of the pedestal condition suggests that the two movements are not fully integrated and, when precision demands are great, remain sensitive to different perception-action constraints in 12-month-old infants.

Background Advances in therapy, surgery and loco-regional procedures improved survival of patients (pts) with metastatic colorectal cancer (mCRC). Interruptions, defined Drug Holidays (DH), could reduce toxicity in pts with stable disease. We evaluated the association between DH and overall survival (OS). Methods 754 consecutive pts treated for mCRC, at University Hospital of Udine and IRCCS CRO of Aviano from 1/1/2005 to 15/03/2017, were included. DH was defined as 56 or more consecutive days of interruption, during first line. The association of DH to OS was evaluated through uni- and multivariate Cox regression analyses. OS was estimated with Kaplan-Meier curves. Results Overall, 459 (60.9%) pts received continuous treatment while 255 (33.8%) a DH (5.3% missing data). After median follow-up of 68.6 months, median OS was 23.15 months. By univariate analysis, KRAS (HR 1.39; 95%CI 1.17-1.66; p 1 metastatic sites (HR 1.61;95%CI 1.36-1.91; p Conclusions DH was independently associated with better prognosis. Probably, DH is a proxy of better prognosis that clinicians intercept. Therefore, in accurately selected pts, DH can be safely offered. Legal entity responsible for the study Dipartimento di Oncologia, Azienda Sanitaria Universitaria Integrata di Udine. Funding Has not received any funding. Disclosure F. Puglisi: Advisory / Consultancy, Speaker Bureau / Expert testimony, Research grant / Funding (institution), Travel / Accommodation / Expenses: Hoffmann-La Roche; Advisory / Consultancy, Speaker Bureau / Expert testimony, Research grant / Funding (institution), Travel / Accommodation / Expenses: AstraZeneca; Honoraria (self), Advisory / Consultancy, Speaker Bureau / Expert testimony, Travel / Accommodation / Expenses: Amgen; Advisory / Consultancy, Speaker Bureau / Expert testimony, Travel / Accommodation / Expenses: Celgene; Advisory / Consultancy, Speaker Bureau / Expert testimony, Travel / Accommodation / Expenses: Eisai; Advisory / Consultancy, Speaker Bureau / Expert testimony, Travel / Accommodation / Expenses: Eli Lilly; Honoraria (self), Advisory / Consultancy, Speaker Bureau / Expert testimony, Travel / Accommodation / Expenses: Ipsen; Advisory / Consultancy, Speaker Bureau / Expert testimony, Travel / Accommodation / Expenses: Novartis; Honoraria (self), Advisory / Consultancy, Speaker Bureau / Expert testimony, Travel / Accommodation / Expenses: Pierre-Fabre; Advisory / Consultancy, Speaker Bureau / Expert testimony, Travel / Accommodation / Expenses: Pfizer; Advisory / Consultancy, Speaker Bureau / Expert testimony, Travel / Accommodation / Expenses: Takeda; Advisory / Consultancy: Merck Sharp & Dohme. All other authors have declared no conflicts of interest.

Background Cancer multidisciplinary team meetings (MDMs) are commonly acknowledged as a good clinical practice. One of the roles of MDMs is to identify the best diagnostic and therapeutic strategies for patients (pts) with new diagnosis of early breast cancer (EBC). In this setting, the purpose of the study was to define whether there was agreement between the planned program (i.e. MDMs-based decision) and that actually applied (i.e. actual therapeutic choice, ATC). In addition, the study explored factors associated with discordance. Methods We conducted a monocentric retrospective study of a consecutive series of 291 pts with new diagnosis of EBC, discussed at MDMs at the University Hospital of Udine (Italy), from January 2017 to June 2018. Results Median age was 62 years (range 27-88 years). Among invasive EBC patients, the most frequent phenotype was luminal-A (38%), followed by luminal-B (33%), HER2-positive (12%) and triple negative (5%). Thirty-four pts (12%) had diagnosis of in situ carcinoma (DCIS). Median time from MDMs discussion to first oncological examination was two weeks. Rate of discordance between MDMs-based decision and final choice, during face to face consultation with the oncologist, was 15.8% (46/291). Among cases with discordance, 19 pts (41.3%) had age > 70 years; 8 pts (17%) had a diagnosis of DCIS, 13 pts (28%) luminal-B carcinoma, 12 pts (26%) luminal-A, 9 pts (20%) HER2-positive and 4 pts (9%) triple negative EBC. The most frequent reason for changing the MDMs-based program was clinical decision by the oncologist at the first evaluation (87%). Follow-up was preferred to the chemotherapy proposed within the MDMs by 15% of pts, and to the endocrine therapy in 39% cases (among these, 44.5% had diagnosis of DCIS). In our study 16/46 pts (35%) had a therapeutic change from chemotherapy to endocrine therapy: among these pts, 7/16 had a luminal-B and 6/16 had a HER2-positive disease. Further analysis aiming at evaluating variables which could predict discordance between MDMs proposal and face to face oncological consultation are ongoing. Conclusions The results of our study could be useful for enhance the role of MTD and identify unmet needs in decision making process in EBC. Legal entity responsible for the study The authors. Funding Has not received any funding. Disclosure All authors have declared no conflicts of interest.

Background High BMI is associated with better survival in metastatic melanoma patients (MM pts), while sarcopenia is linked to poorer outcome in pts with stage III. The aim of this study was to examine the prognostic impact of body-mass index (BMI), baseline sarcopenia, loss of skeletal muscle mass (LSMM) on overall survival (OS) in MM pts who received immunotherapy (IT). Methods The retrospective series included 42 consecutive MM pts (Jan 2011-Dec 2018) treated with IT in a single referral center. Sarcopenia was defined according to Prado’s criteria. Skeletal muscle index (SMI) was calculated as cross-sectional-area of muscle (cm2), using CT-scan, at the L3 level divided by the square of the height (m2). Early LSMM, during IT, was defined as a decrease in SMI > =10% from baseline at first evaluation. BMI was calculated as weight (kg) divided by the square of height (m2) and categorized according to standard WHO definitions. Weight loss was analyzed as continuous variable. Results At baseline, 27 pts (64,3%) were male, 26 pts (61.9%) were =25. Out of 42 pts, 30 (71.4%) had a CT-scan at first evaluation, and 30% of them had an early LSMM. Median OS was 11.38 months. Both in univariate and multivariate analysis, ECOG PS > =1, and early LSMM > =10% were significantly associated with worse OS. Conversely, pts with weight loss had better OS (Table). Table . 1367P OS univariate analysis OS multivariate analysis Factors HR p 95% Confidence Interval HR p 95% Confidence Interval PS ECOG³1 3.02 0.01 (1.29-7.08) 3.99 0.043 (1.04-15.31) Weight loss 0.88 0.03 (0.78-0.98) 0.85 0.02 (0.74-0.97) Early LSMM > =10% 4.24 0.006 (1.50-11.97) 3.09 0.04 (1.04-9.22) Conclusions Early LSMM > =10% and ECOG PS > =1 may negatively influence the outcome of MM pts treated with IT. Further prospective studies are needed to confirm these data. Legal entity responsible for the study Maria Grazia Vitale. Funding Has not received any funding. Disclosure All authors have declared no conflicts of interest.

Background Patients (pts) with melanoma brain metastases (BM) have a historically poor prognosis, with a reported median overall survival (mOS) of 4-6 months. Aim of this study is to evaluate predictive factors of survival in pts with melanoma BM. Secondly, a multivariate model was used to define groups with distinct prognosis. Methods Consecutive pts with melanoma BM treated in two Italian cancer centers were included in this 10-year retrospective cohort study (2008-2018). Baseline clinicopathological factors, systemic and loco-regional treatments were registered. The Kaplan-Meier method and the Cox model were used to analyze the association of prognostic factors and OS. Results 150 pts with melanoma BM were enrolled in this study: 42% with a single BM, 51.3% with ≥ 3 extracranial metastatic sites and 38% with an ECOG performance status (PS) of 0. About 50.7% of pts had a BRAF mutation. Notably, 56% of pts were asymptomatic at BM diagnosis, and 40% presented high LDH levels at baseline. Overall, mOS after BM diagnosis was 5.78 months (95% CI 4.80-6.67), and median intracranial progression-free survival was 4.63 months (95% CI 3.81-5.12). According to treatment modality, stereotactic radiosurgery/neurosurgery plus systemic therapy showed the best outcome (mOS 12.32 months, 95% CI 7.92-25.30). A multivariate model was used to identify baseline prognostic factors independently associated with OS (Table). Conversely, sex and symptomatic BM did not significantly impact on OS. Predicted survival functions defined 3 distinct prognostic groups (P Table . 1363P Covariates HR 95% CI P Points BRAF wild type 1.95 1.22-3.12 1 BM > 1 2.20 1.35-3.59 1 LDH ≥ 2 x upper normal limit 1.83 1.07-3.11 0.026 1 ECOG PS > 0 2.68 1.62-4.43 0.0001 1 Extracranial metastatic sites ≥ 3 1.91 1.17-3.1 1 Age ≥ 65 1.86 1.15-3.02 0.012 1 Conclusions This score can be useful for stratifying prognosis in pts with melanoma BM. Prospective studies are needed to validate this score and to explore how it may guide treatment strategy. Legal entity responsible for the study Dipartimento di Oncologia, Azienda Sanitaria Universitaria Integrata di Udine. Funding Has not received any funding. Disclosure F. Puglisi: Honoraria (self), Advisory / Consultancy: Amgen; Advisory / Consultancy: Celgene; Advisory / Consultancy: Eisai; Advisory / Consultancy: Eli Lilly; Honoraria (self), Advisory / Consultancy: Ipsen; Honoraria (self), Research grant / Funding (self): Novartis; Advisory / Consultancy: Pfizer; Advisory / Consultancy, Research grant / Funding (self): Roche; Honoraria (self): Pierre Fabre; Advisory / Consultancy: Merck Sharp & Dohme; Advisory / Consultancy: Takeda; Research grant / Funding (self): Astrazeneca. All other authors have declared no conflicts of interest.

e24014Background: The role of serum tumor markers (TM), such as Ca15-3 and CEA, in early intercepting breast cancer (BC) recurrences is still debated. Due to lack of sensitivity and specificity, TM...

To promote the diffusion of smart metering, several National Authorities defined strict roll-out procedures of the existing meters with new smart meters. In such scenario, even if new static technologies in gas metering offer very significant potential (such as digital output, absence of moving parts, direct mass measurement), their reliability in some critical conditions typical of natural gas distribution networks (i.e. zero flow, minimum flow-rate, gas quality changes, presence of dust and contaminants, etc.) still has to be demonstrated. A very promising technology for natural gas domestic measurements is represented by thermal mass flow meters, but, unfortunately, gas quality is expected to significantly affect metrological performance of such meters. The authors conducted several metrological tests aimed to verify the sensitivity of such meter with natural gas quality changes. The results of these tests show a not full reliability when CO2 content increases and when gas composition varies.

Methylation in vivo is a post-translational modification observed in several organisms belonging to eucarya, bacteria, and archaea. Although important implications of this modification have been demonstrated in several eucaryotes, its biological role in hyperthermophilic archaea is far from being understood. The aim of this work is to clarify some effects of methylation on the properties of β-glycosidase from Sulfolobus solfataricus, by a structural comparison between the native, methylated protein and its unmethylated counterpart, recombinantly expressed in Escherichia coli. Analysis by Fourier transform infrared spectroscopy indicated similar secondary structure contents for the two forms of the protein. However, the study of temperature perturbation by Fourier transform infrared spectroscopy and turbidimetry evidenced denaturation and aggregation events more pronounced in recombinant than in native β-glycosidase. Red Nile fluorescence analysis revealed significant differences of surface hydrophobicity between the two forms of the protein. Unlike the native enzyme, which dissociated into SDS-resistant dimers upon exposure to the detergent, the recombinant enzyme partially dissociated into monomers. By electrospray mapping, the methylation sites of the native protein were identified. A computational analysis of β-glycosidase three-dimensional structure and comparisons with other proteins from S. solfataricus revealed analogies in the localization of methylation sites in terms of secondary structural elements and overall topology. These observations suggest a role for the methylation of lysyl residues, located in selected domains, in the thermal stabilization of β-glycosidase from S. solfataricus

The scallop Adamussium colbecki can be profitably used for monitoring Antarctic coastal environments but its utility would be increased if chemical analyses of pollutants were integrated with data on their biological effects. Since oxidative stress is a common pathway of toxicity induced by xenobiotics, a preliminary biochemical characterization was carried out on the antioxidant system of this species and baseline data collected for future assessment of the anthropogenic impact in this remote area. The digestive gland and gills were investigated for levels of glutathione and the activity of several glutathione-dependent and antioxidant enzymes: gluthathione reductase, EC 1.6.4.2; glyoxalase I, EC 4.4.1.5; glyoxalase II, EC 3.1.2.6; gluthathione S-transferases, EC 2.5.1.18; Se-dependent, EC 1.11.1.9 and Se-independent, EC 2.5.1.18 gluta-thione peroxidases; catalase, EC 1.11.1.6; and super-oxide dismutase, EC 1.15.1.1. The same enzymatic activities were measured for comparison in the Mediterranean molluscs Mytilus galloprovincialis and Pecten jacobaeus. Very high levels of glutathione S-transferases were found in the digestive gland of both species of scallop compared to mussels, suggesting the importance of different feeding behaviour among these molluscs. However, catalase activity, much higher in Adamussium colbecki than in the Mediterranean molluscs, may represent a biochemical adaptation to the Antarctic marine environment with high levels of dissolved oxygen. Enzymes from the Antarctic species appeared to be generally more active at low temperatures but, with a few exceptions, their activities increased at higher temperatures.

L'articolo illustra i risultati derivanti dalla somministrazione di 451 questionari a pazienti affetti da Human Immunodeficiency Virus (HIV) ricoverati in reparti di malattie infettive appositamente attrezzati per la cura dell'AIDS o afferenti ai relativi ambulatori o day-hospital. In particolare, l'indagine ha inteso verificare tra le persone intervistate l'effettiva conoscenza, comprensione, valutazione ed applicazione degli articoli 5 e 6 della legge italiana n°135 del 1990 su "Programma di interventi urgenti per la prevenzione e la lotta contro l'AIDS". Tali articoli, infatti, riguardano le normative sull'accertamento dell'infezione (art. 5) e il divieto per i datori di lavoro di svolgere indagini volte ad accertare la sieropositività dei propri dipendenti (art.6). L'indagine è stata condotta in tre ospedali: Centro S. Luigi dell'Istituto Scientifico Ospedale S. Raffaele di Milano (150 questionari), Policlinico S. Matteo di Pavia (150 questionari) e Policlinico Universitario "A. Gemelli" di Roma (150 questionari). Il questionario è stato strutturato sulla base di doppie domande: una su ciò che il paziente personalmente pensa riguardo ai suoi "diritti", l'altra su ciò che il paziente pensa che la legge preveda. In generale sembra emergere che i soggetti intervistati sono coscienti di essere persone "infette", però in una comunità che tende a discriminarli. Non sembrano attendersi, inoltre, una piena tutela da parte dello Stato anche se rimangono propensi a collaborare perché la situazione cambi e sono, comunque, disposti ad accettare interventi normativi su se stessi a condizione della non discriminazione e che il loro sacrificio risulti veramente utile per la prevenzione della diffusione del contagio. Risalta, infine, l'esigenza di mantenere riservato lo stato di malattia nel luogo di lavoro.

Many drugs and chemicals have been shown to induce modifications of the physicochemical properties of cellular membranes. In this study we investigated the changes in fluidity of erythrocyte membrane from epileptic patients under different pharmacological treatments, with respect to healthy controls, by using trimethylammonium-1,6-diphenyl-1,3,5-hexatriene (TMA-DPH) fluorescence polarization. The increase in TMA-DPH fluorescence polarization values observed in epileptic patients indicated a decrease in membrane fluidity. Since the analysis of erythrocyte membrane composition did not reveal significant differences between the two groups studied, a correlation with membrane lipoperoxide content was tried, as different drugs and chemicals elicit in vivo alterations resulting in peroxidation of membrane lipids. Therefore the presence of peroxidation products in the blood and the possible correlation with membrane lipoperoxide were studied. Although a direct causal linkage cannot be proved we can hypothesize that exogenous compounds such as antiepileptic drugs could modify membrane fluidity by increasing membrane lipid peroxidation. Moreover the increase of peroxidative products in the blood could indicate that the peroxidative damage might propagate through the formation of new free radical species. The possibility of using erythrocyte membrane as a model system to analyze antiepileptic drug side effects is advanced.

The distance between the beta-subunits of Na+/K+-ATPase isolated from pig dark red kidney medulla was determined by Förster energy transfer. First, oligosaccharides of the beta-subunit were shown to be labelled with three fluorophores: Lucifer yellow (LY), Lissamine rhodamine B sulfonyl hydrazine (LRSH) and Cascade blue (CB). Further, LY and LRSH were used as the donor and the acceptor, respectively, for Förster energy transfer studies to determine the localization of the beta-subunit in the native enzyme which is known to be formed as a tetramer (alphabeta)2. It was found that the beta-subunits in the functional enzyme complex in the membrane are not localized next to each other but are spatially separated. The distance between fluorophores covalently attached to the beta-subunits was found to be 5.1 nm. This conclusion was confirmed by measurements with another donor-acceptor pair CB-LY. The results also support the idea of a direct interaction of the beta-subunit with the extracellular part of the alpha-subunit. These interactions were modified in the presence of millimolar concentrations of magnesium ions. This indicates a crucial role of magnesium in extracellular interactions between the alpha and beta subunits.

Since a general pathway of toxicity induced by pollutants is the enhancement of reactive oxygen species, biochemical responses associated with variations in the antioxidant cellular system have been often proposed as biomarkers of pollutant-mediated toxicity associated with oxidative stress. Antarctic organisms live in an extreme environment characterized by low water temperature, high level of dissolved oxygen, presence of ice and strong seasonal changes in light intensity and availability of food, conditions which could influence both the formation of reactive oxygen species and the mechanisms for their removal. In this respect and considering the utility of this as a key species for monitoring marine Antarctic environment it was of interest to investigate the antioxidant defense system of the scallop Adamussium colbecki.

Fluorescence emission intensity of 1,3-diphenylisobenzofuran incorporated in polymorphonuclear granulocytes plasma membranes was investigated in basal conditions and during stimulation with different activators. Phorbol myristate acetate, known as the most effective "oxygen burst" inducer, produced a larger decrease in 1,3-diphenylisobenzofuran fluorescence intensity than 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine (platelet activating factor) and N-formyl-methionyl-leucyl-phenylalanine, known as weak stimulants of oxygen uptake. Diphenyl iodonium an inhibitor of leukocyte NADPH oxidase, and the singlet oxygen selective trap alpha-terpinene inhibited the quenching effect of phorbol myristate acetate on 1,3-diphenylisobenzofuran fluorescence. These data suggest formation of singlet oxygen in activated leukocytes and demonstrate that measurement of 1,3-diphenylisobenzofuran fluorescence intensity provides a new sensitive method of detection of neutrophils activation.

Seaprose is a semialkaline proteinase endowed with proteolytic effect and antiinflammatory activity tested in different clinical trials. There is clinical evidence that seaprose reduces sputum viscoelastic properties in chronic hypersecretory bronchitis. The present study evaluated (in a double-blind design vs. placebo) the activity of seaprose on bronchial inflammation, mucus glycoprotein secretion and bronchial humoral defence mechanism in chronic bronchitic patients clinically stable (10 per group). Markers of bronchial inflammation (albumin, albumin/total protein ratio) and bronchial infection (DNA), of mucus glycoproteins (fucose and N-acetylneuraminic acid) and of humoral defence mechanism (secretory-IgA) were tested in sputum. We found that ten-day treatment with seaprose (90 mg/day) reduced sputum albumin during the observation period, the difference being statistically significant at the 18th day. The sputum albumin/total protein ratio also decreased by 50% at the end of the study. In the same group, sputum DNA, secretory-IgA, fucose and N-acetylneuraminic acid remained unchanged after treatment. The placebo group did not show any significant changes in the sputum marker substances. This study provides experimental evidence for the antiinflammatory activity of seaprose on bronchial mucosa in chronic bronchitic patients studied in a stable phase of their disease. Furthermore the drug does not seem to affect mucus glycoprotein secretion or secretory-IgA production.

A new cuvette was developed for spectrofluorometric measurements which increases the efficiency of recorded fluorescence emission by at least two-fold, decreasing, at the same time, the noise component of the record. It is suited for both steady-state and dynamic fluorescence measurements. Comparative data for both the classical and the novel cuvettes are presented here.

Human erythrocytes were separated using Percoll/Albumin density gradient centrifugation into 5 subpopulations which represent cell subpopulations of different ages. The physico-chemical properties of the membranes and their lipid extracts, prepared from the 5 subpopulations, were studied by 1,6-diphenyl-1,3,5-hexatriene fluorescence polarization and by Fourier Transform Infrared Spectroscopy respectively. The fluorescence polarization data show a decrease of membrane fluidity in the youngest erythrocyte fraction and the infrared spectroscopy reveals a lower degree of unsaturation of the lipids from this fraction. Changes of membrane fluidity among the 5 erythrocyte subpopulations is assigned to the variations of the degree of unsaturations of lipids rather than to variations of cholesterol content.

1. Mitochondria isolated from porcine adrenal cortex under State 3 conditions oxidized succinate with a rate of 47 +/- 4.48 na oxygen/min/mg/protein and with ADP:O ratio 0.98 +/- 0.09. In the presence of 15 microM deoxycorticosterone the rate of succinate oxidation was 36.8 +/- 3.08 na oxygen/min/mg/protein. 2. Under the same conditions the rate of glutamate oxidation was 22.8 +/- 2.21 and 16.8 +/- 0.65 na oxygen/min/mg/protein, respectively. ADP:O ratio was 1.45 +/- 0.14. 3. Introduction of trace amounts of malate into the mitochondria oxidizing glutamate only slightly increased the rate of O2 uptake. 4. The glutamate dehydrogenase activity in these mitochondria was 12.5 +/- 0.69 nmol/min/mg.

Acute lung injury/acute respiratory distress syndrome (ALI/ARDS) is characterized by an important inflammatory reaction involving both lungs, often spreading systemically. Pentraxin 3 (PTX3) has recently been described as a mediator of the acute phase of innate immunity. It belongs, together with C-reactive protein (CRP), to the pentraxin superfamily. PTX3 is produced by endothelial cells, fibroblasts, mononuclear phagocytes and dendritic cells in response to primary inflammatory signals (IL-1, tumor necrosis factor, LPS). Its involvement in the pathogenesis of ALI/ARDS seems probable and potentially interesting for the pathophysiology, the diagnosis and possibly the therapy of the syndrome. We measured PTX3 levels, by an ELISA test, in the sera of 25 patients affected by ALI/ARDS. We also collected data related to etiology, pulmonary mechanics, gas exchange, systemic inflammatory involvement, microbiology, blood cells and the outcome of these patients. PTX3 was markedly elevated in all 25 patients (average 111.07 ± 192.01 ng/ml; range 4.94–922.91 ng/ml). In no patient were PTX3 levels lower or equal to the physiological threshold of 2 ng/ml. Levels of logPTX3 were significantly different between survivor and nonsurvivor patients (1.465 ± 0.529 log ng/ml vs 1.950 ± 0.643 log ng/ml; P = 0.05). Patients with two or more organ failures (besides the lung) also had logPTX3 levels significantly higher than patients with just one additional organ failure (1.936 ± 0.597 log ng/ml vs 1.423 ± 0.542 log ng/ml; P < 0.04). The same was true when PTX3 levels in patients with septic shock were compared with PTX3 levels in patients without shock (262.465 ± 350.727 ng/ml vs 63.257 ± 70.261 ng/ml; P < 0.03). PTX3 and logPTX3 were correlated with body temperature (r = 0.56 P < 0.01; r = 0.51 P = 0.01) and with arterial blood pH (r = -0.53 P < 0.01; r = -0.53 P < 0.01). We have underlined a relationship between circulating levels of PTX3 and some markers of the severity of systemic involvement. PTX3 could be an indicator of the gravity of the systemic inflammatory reaction and of unfavourable outcome in ALI/ARDS patients.