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Background: Triple Negative Breast Cancers (TNBC) represent 12% to 20% of total Breast Cancers (BC) and have a worse outcome compared with other breast cancer subtypes. TNBC often show a deficiency in DNA double strand break repair mechanisms. This deficiency is generally related to inactivation of a repair enzymatic complex involving BRCA1 caused either by genetic mutations, epigenetic modifications, or by post-transcriptional regulations. The identification of BC presenting a BRCA1 deficiency could be useful to select patients that could benefit from PARP inhibitors, alkylant agents or platinum-based chemotherapy. In this study, we have identified by Comparative Genomic Hybridization array (CGH-array) a recurrent gain in 17q25.3 characteristic of BRCA1 mutated or methylated TNBC. Methods: 130 formalin-fixed paraffin embedded (FFPE) tumours including TNBC (unknown-BRCA1-status, BRCA1 mutated and non-mutated), Luminal A, Luminal B, Her2-neu amplified (Her2+) BC and BRCA1-mutated non-TNBC (mutated-Luminal A, Luminal B, Her2+) were obtained from our local tumour collection. DNA was extracted and genomic copy-number alterations were analysed by CGH-array (Agilent, SuperPrint G3 Human CGH 8×60K Oligo Microarrays). FISH analyses were performed on section from FFPE samples to validate some chromosomal aberrations belonging to the 17q25.3 amplified region evidenced by CGH-array. The study of BRCA1 promoter methylation status in all tumours was carried out by MDxHealth (Liege, Belgium). Results: In this study, we have identified by CGH-array a genomic region (17q25.3) amplified in 90% of the BRCA1 mutated tumours (29/32). This chromosomal gain was studied in other subtypes of BC by CGH-array and it was only evidenced in 30% (6/20) of BRCA1 non-mutated TNBC, 26.67% (4/15) of unknown-BRCA1-status TNBC, 13.64% (3/22) of Luminal B, 19.05% (4/21) of Her2+ and 0% (0/20) of Luminal A breast cancers. FISH assays confirmed these chromosomal amplifications and evidenced like CGH array analyses a significant difference between BRCA1 mutated and non-mutated BC for the 17q25.3 gain. BRCA1 methylation was found only in TNBC (11/58) and was not found in the BRCA1-mutated BC cohort, nor in the Luninal A, Luminal B and Her2+ samples. In BRCA1 non-mutated TNBC, the methylation was found in 8 cases including 4 with 17q25.3 amplification. In the unknown-BRCA1-status TNBC, 3 methylated samples were found with 1 case with co-amplification of the 17q25.3 region. Recurrence of 17q25.3 amplification in BRCA1 mutated tumours as well as its detection in BC having a methylation in BRCA1 promoter suggests that the 17q25.3 gain could be a marker of the BRCA1 deficiency. Identification of relevant genes whose expression is up-regulated within the recurrently gained region is underway. Conclusions: The CGH signature observed in 17q25.3 chromosomal region and FISH assay developed in this study could allow the identification of “BRCAness” breast tumours, improving the diagnostic performance and orienting the selection of the appropriate therapy. The up-regulated genes themselves might also represent potential therapeutic targets. Acknowledgements: This work was financed through the “Plan National Cancer-Action 29” (Belgium) and supported by the UNICANCER-PACS08 trial (France). Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-04-04.

Background Irinotecan (IR) or oxaliplatin (OX)-based regimens optionally combined with anti-VEGF or anti-EGFR Target Therapies (TT) are used as preoperative treatment for metastatic colorectal patients before resection of CRCM. Best combination remains unclear. The study purpose was to compare PR observed on resected CRCM after different preoperative treatments. Methods 114 patients engaged for CRCM resection after 2005 were included in this retrospective analysis. PR was evaluated on 296 resected CRCM according to the pathological Tumor Regression Grading scale (TRG), grading PR from complete (TRG1) to absent (TRG5). Mean TRG of resected metastasis was compared based on Kruskall-Wallis and Mann-Whitney tests. Cumulative PFS were calculated by Kaplan-Meir method and compared by log-rank test. Results 92/114 patients were preoperatively treated. Mean TRG after OX without TT was better than mean TRG after IR without TT (p = .044). No difference was observed between mean TRG after chemotherapy alone compared to mean TRG after chemotherapy + anti-VEGF (p = .53) or to mean TRG after chemotherapy + anti-EGFR (p = .39). Subgroup analysis revealed that mean TRG after IR + anti-EGFR was better than mean TRG after OX + anti-EGFR (p = .031), and that mean TRG after OX + anti-VEGF was better than mean TRG after IR + anti-VEGF (p = .001). PFS analysis for the 92 preoperatively treated patients revealed that 13 patients with major PR (TRG ≤ 2), had a significantly improved PFS compared to 79 patients with minor or no PR (TRG > 2) (median PFS= 33.7 vs 22.9 month p= .018). Conclusion PR observed on resected CRCM after a preoperative treatment does not seem to be linked to the TT (anti-VEGF or anti-EGFR) but rather to the chemotherapy/TT association with a significant advantage for OX + anti-VEGF or IR + anti-EGFR combinations. Preoperative strategy Patients (n) Metastases (n) TRG mean 95% CI No treatment 22 54 4.33 4.11- 4.55 Chemo alone 38 86 3.19 2.95 - 3.43 OX 28 68 3.16 2.89-3.43 IR 8 11 3.91 3.44-4.38 Other 2 7 ND ND Anti-VEGF + chemo 38 120 3.32 3.10 - 3.54 OX 8 27 2.63 2.18 - 3.08 IR 29 80 3.53 3.27 - 3.78 Other 1 13 ND ND Anti-EGFR + chemo 16 36 2.97 2.50 - 3.44 OX 9 16 3.56 2.98 - 4.15 IR 6 19 2.47 1.77 - 3.18 Other 1 1 ND ND Disclosure All authors have declared no conflicts of interest.

6-Aminonicotinamide (6-AN) is a niacin antagonist, which leads to degeneration of gray-matter astrocytes followed by a vigorous inflammatory response. Macrophage colony stimulating factor (M-CSF) is important during inflammation, and in order to further clarify the roles for M-CSF in neurodegeneration and brain cell death, we have examined the effect of 6-AN on osteopetrotic mice with genetic M-CSF deficiency (op/op mice). The 6-AN-induced degeneration of gray-matter areas was comparable in control and op/op mice, but the numbers of reactive astrocytes, macrophages, and lymphocytes in the damaged areas were significantly decreased in op/op mice relative to controls. The levels of oxidative stress (as determined by using immunoreactivity for inducible nitric oxide synthase, nitrotyrosine, and malondialdehyde) and apoptotic cell death (as determined by using TUNEL and immunoreactivity for caspases and cytochrome c) were significantly increased in 6-AN-injected op/op mice relative to controls. From a number of antioxidant factors assayed, only metallothioneins I and II (MT-I+II) were decreased in op/op mice in comparison to controls. Thus, the present results indicate that M-CSF is an important growth factor for coping with 6-AN-induced central nervous system damage and suggest that MT-I+II are likely to have a significant role.

Metyrapone, a 11-beta steroid hydroxylase inhibitor that blocks stress-induced glucocorticoid release, is extensively used to study the physiological and behavioural roles of glucocorticoids. However, there is circumstantial evidence suggesting that metyrapone could act as a pharmacological stressor. Thus, the effects of various doses of metyrapone on two well-characterized stress markers (ACTH and glucose) were studied in male rats. Metyrapone administration, while exerting a modest effect on plasma corticosterone levels, dose-dependently increased plasma ACTH and glucose levels. Using the highest doses previously tested (200 mg/kg) we further observed, as evaluated by fos-like immunoreactivity (FLI), a strong activation of a wide range of brain areas, including the parvocellular region of the hypothalamic paraventricular nucleus (PVNp), the origin of the main ACTH secretagogues. Metyrapone-induced FLI was observed in neocortical and allocortical areas, in several limbic, thalamic and hypothalamic nuclei and, to a lesser extent, in the brainstem. In a final experiment, a dose-response study of metyrapone-induced FLI was carried out focusing on selected brain areas. The study revealed that the paraventricular thalamic nucleus and central amygdala were the areas most sensitive to metyrapone as they responded even to the lowest dose of the drug. Most areas, among them the PVNp, only showed enhanced FLI with the two highest doses, i.e. when it was associated with ACTH and glucose responses. These data suggest that some of the effects of metyrapone could be due to its stressful properties rather than its ability to inhibit glucocorticoid synthesis. The exact mechanisms involved remain to be established.

Publisher Summary This chapter describes metallothionein (MT) expression in the brain. MTs constitute a family of protein with a high affinity for heavy metals, such as cadmium and zinc, and an unusual cysteine content in peripheral tissues. Based on structural relationships, MTs have been subdivided into several families. Mammalian MTs belong to family l, which is composed of several subfamilies, ml to m4. In the mouse, ml to m4 subfamilies are composed of a single member (MT-1 in the ml subfamily, MT-2 in the m2 subfamily, etc.), which is in contrast to primates where a significant genetic polymorphism exists. Metallothioneins-1 and -2 (MT-1 +2) are expressed coordinately in most tissues, including the central nervous system (CNS), whereas MT-3 is primarily confined to the CNS and MT-4 to the stratified squamous epithelia.

Methanol is a versatile chemical feedstock, fuel source, and energy storage material. Many reactions involving methanol are catalyzed by transition metal surfaces, on which hydrogen-bonded methanol overlayers form. As with water, the structure of these overlayers is expected to depend on a delicate balance of hydrogen bonding and adsorbate-substrate bonding. In contrast to water, however, relatively little is known about the structures methanol overlayers form and how these vary from one substrate to another. To address this issue, herein we analyze the hydrogen bonded networks that methanol forms as a function of coverage on three catalytically important surfaces, Au(111), Cu(111), and Pt(111), using a combination of scanning tunneling microscopy and density functional theory. We investigate the effect of intermolecular interactions, surface coverage, and adsorption energies on molecular assembly and compare the results to more widely studied water networks on the same surfaces. Two main factors are shown to direct the structure of methanol on the surfaces studied: the surface coverage and the competition between the methanol-methanol and methanol-surface interactions. Additionally, we report a new chiral form of buckled hexamer formed by surface bound methanol that maximizes the interactions between methanol monomers by sacrificing interactions with the surface. These results serve as a direct comparison of interaction strength, assembly, and chirality of methanol networks on Au(111), Cu(111), and Pt(111) which are catalytically relevant for methanol oxidation, steam reforming, and direct methanol fuel cells.

The role of interleukin-6 in hippocampal tissue damage after injection with kainic acid, a rigid glutamate analogue inducing epileptic seizures, has been studied by means of interleukin-6 null mice. At 35mg/kg, kainic acid induced convulsions in both control (75%) and interleukin-6 null (100%) mice, and caused a significant mortality (62%) only in the latter mice, indicating that interleukin-6 deficiency increased the susceptibility to kainic acid-induced brain damage. To compare the histopathological damage caused to the brain, control and interleukin-6 null mice were administered 8.75mg/kg kainic acid and were killed six days later. Morphological damage to the hippocampal field CA1-CA3 was seen after kainic acid treatment. Reactive astrogliosis and microgliosis were prominent in kainic acid-injected normal mice hippocampus, and clear signs of increased oxidative stress were evident. Thus, the immunoreactivity for inducible nitric oxide synthase, peroxynitrite-induced nitration of proteins and byproducts of fatty acid peroxidation were dramatically increased, as was that for metallothionein I+II, Mn-superoxide dismutase and Cu/Zn-superoxide dismutase. In accordance, a significant neuronal apoptosis was caused by kainic acid, as revealed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling and interleukin-1beta converting enzyme/Caspase-1 stainings. In kainic acid-injected interleukin-6 null mice, reactive astrogliosis and microgliosis were reduced, while morphological hippocampal damage, oxidative stress and apoptotic neuronal death were increased. Since metallothionein-I+II levels were lower, and those of inducible nitric oxide synthase higher, these concomitant changes are likely to contribute to the observed increased oxidative stress and neuronal death in the interleukin-6 null mice. The present results demonstrate that interleukin-6 deficiency increases neuronal injury and impairs the inflammatory response after kainic acid-induced seizures.

Transgenic mice expressing TNF-alpha under the regulatory control of the GFAP gene promoter (GFAP-TNFalpha mice) exhibit a unique, late-onset chronic-progressive neurological disorder with meningoencephalomyelitis, neurodegeneration, and demyelination with paralysis. Here we show that the metallothionein-I + II (MT-I + II) isoforms were dramatically upregulated in the brain of symptomatic but not presymptomatic GFAP-TNFalpha mice despite TNF-alpha expression being present in both cases. In situ hybridization analysis for MT-I RNA and radioimmunoassay results for MT-I + II protein revealed that the induction was observed in the cerebellum but not in other brain areas. Increased MT-I RNA levels occurred in the Purkinje and granular neuronal layers of the cerebellum but also in the molecular layer. Reactive astrocytes, activated rod-like microglia, and macrophages, but not the infiltrating lymphocytes, were identified as the cellular sources of the MT-I + II proteins. In situ hybridization for MT-III RNA revealed a modest increase in the white matter of the cerebellum, which was confirmed by immunocytochemistry. MT-III immunoreactivity was present in cells which were mainly round or amoeboid monocytes/macrophages. The pattern of expression of the different MT isoforms in the GFAP-TNFalpha mice differed substantially from that described previously in GFAP-IL6 mice, demonstrating unique effects associated with the expression of each cytokine. The results suggest that the MT expression in the CNS reflects the inflammatory response and associated damage rather than a direct role of the TNF-alpha in their regulation and support a major role of these proteins during CNS injury.

The balance between trophic factors and inhibitory molecules is likely to determine the outcome of neural tissue damage. The growth inhibitory factor (GIF), a member of the metallothionein family of proteins named metallothionein-III (MT-III), has been suggested to play an important role in tissue repair after adult brain injury. Because no information is available on this factor in relation to immature brain damage, we examined the chronological changes of GIF (MT-III) mRNA and protein following excitotoxic lesions to the postnatal day 9 brain using in situ hybridization and immunocytochemical techniques. We observed a significant decrease of neuronal GIF (MT-III) mRNA and protein levels between 4 and 24 hours postinjury and an increase in glial GIF (MT-III) levels. Double immunocytochemical techniques showed GIF (MT-III) and GFAP positive astrocytes from 2-4 hours postinjury. From 3 days postinjury strongly reactive astrocytes expressed strong levels of both GIF (MT-III) mRNA and protein, which were maintained in the glial scar formed at longer times. These results show the expression of an inhibitory molecule by postnatal reactive astrocytes. Glial GIF (MT-III) expression may play an important role in the tissue reconstruction after immature brain damage.

The role of nitric oxide (NO) production on metallothionein (MT) regulation in the liver and the brain has been studied in mice by means of the administration of nitric oxide synthase (NOS) inhibitors. Mice injected with either the arginine analog N G -monomethyl-L-arginine (L-NMMA) or the heme binding compound 7-nitro indazole (7-NI) showed consistently increased liver MT-I mRNA and MT-I+II total protein levels, suggesting that NO is involved in the hepatic MT regulation. In agreement with the liver results, in situ hybridization analysis demonstrated a significant upregulation of the brain MT-I isoform in areas such as the cerebrum cortex, neuronal CA1-CA3 layers and dentate gyrus of the hippocampus, and Purkinje cell layer of the cerebellum, in 7-NI treated mice. The same trend was observed for the brain specific isoform, MT-III, but to a much lower extent. The effect of NOS inhibition was also evaluated in a MT-inducing condition, namely during immobilization stress. In both the liver and the brain, stress upregulated the MT-I isoform, and 7-NI significantly reduced or even blunted the MT-I response to stress, suggesting a mediating role of NO on MT-I regulation during stress. Stress also increased the MT-III mRNA levels in some brain areas, an effect blunted by the concomitant administration of 7-NI, which in some areas even decreased MT-III mRNA levels below the saline injected mice. Results in primary culture of neurons and astrocytes demonstrate significant effects of the NOS inhibitors in some experimental conditions. The present results suggest that NO may have some role on MT regulation in both the liver and the brain.

The effect of interleukin-6 (IL-6) on metallothionein-I (MT-I) and MT-III expression in the brain has been studied in transgenic mice expressing IL-6 under the regulatory control of the glial fibrillary acidic protein gene promoter (GFAP-IL6 mice), which develop chronic progressive neurodegenerative disease. In situ hybridization analysis revealed that GFAP-IL6 (G16-low expressor line, and G36-high expressor line) mice had strongly increased MT-I mRNA levels in the cerebellum (Purkinje and granular layers of the cerebellar cortex and basal nuclei) and, to a lesser degree, in thalamus (only G36 line) and hypothalamus, whereas no significant alterations were observed in other brain areas studied. Microautoradiography and immunocytochemistry studies suggest that the MT-I expression is predominantly localized to astrocytes throughout the cerebrum and especially in Bergman glia in the cerebellum. However, a significant expression was also observed in microglia of the GFAP-IL6 mice. MT-III expression was significantly increased in the Purkinje cell layer and basal nuclei of the cerebellum, which was confirmed by Northern blot analysis of poly(A)1 mRNA and by ELISA of the MT-III protein. In contrast, in the G36 but not G16 mice, transgene expression of IL-6 was associated with significantly decreased MTIII RNA levels in the dentate gyrus and CA3 pyramidal neuron layer of the hippocampus and, in both G36 and G16 mice, in the occipital but not frontal cortex and in ependymal cells. Thus, both the widely expressed MT-I isoform and the CNS specific MT-III isoform are significantly affected in a MT isoform- and CNS area-specific manner in the GFAP-IL6 mice, a chronic model of brain damage. r 1998 Academic Press

Metallothioneins (MTs) are a family of low molecular weight proteins which in rodents is comprised of several isoforms (MT-I to MT-IV). MT-I and MT-II are widely expressed isoforms, whereas MT-III is mainly expressed in the central nervous system and is the only isoform that inhibits survival and neurite formation of rat cortical neurons in vitro. However, the physiological roles and regulation of these proteins in the brain are poorly characterized. In this report we have studied the putative role of IL-6 and TNF-alpha on the regulation of brain MT-I and MT-III, by using mice carrying a null mutation in the IL-6 or the TNF-alpha type 1 receptor genes or both. In situ hybridization analysis revealed that brain MT-I induction by bacterial lipopolysaccharide (LPS) was significantly lower in IL-6- and TNFR1-deficient mice, and to a greater extent in the double mutant mice, in most brain areas studied. These results suggest that the MT-I isoform could be considered an acute-phase protein in the brain, which is consistent with previous studies in transgenic mice overexpressing IL-6 in astrocytes. In contrast to LPS, brain MT-I induction by restraint stress was not affected significantly by IL-6 or TNFR1 deficiencies, suggesting that these cytokines are not important during the stress response in the brain. In basal conditions, it was also observed that the double mutant mice had diminished MT-I mRNA levels in several brain areas. In contrast to MT-I, MT-III mRNA levels were minimally affected by either LPS or stress. Yet, significant decreasing effects of IL-6 and TNFR1 deficiencies were observed in the Purkinje neuronal layer of the cerebellum (after LPS) and ependymal cells (after LPS and stress). In contrast, significant increasing effects, especially of TNFR1 deficiency, were observed in CA1 hippocampal area, retrosplenial and parietal cortex, and in thalamic nuclei (after LPS). These results demonstrate that IL-6 and TNF-alpha are involved in brain MTs regulation during LPS-elicited inflammatory response but not during the stress response.

This paper summarizes experience and lessons learned in the framework of the MEDAS research project. The MEDAS project was devoted to the elaboration of methodological guidelines for the design of communication systems based on the use of FDTs, particularly LOTOS and SDL, and their relationship with object oriented techniques and industrial-strength software development methods.

Background Neoadjuvant chemoradiotherapy (NCR) followed by radical resection is the standard of care for patients with stage II-III rectal cancer. L-BLP25 (Stimuvax®) is an antigen-specific cancer immunotherapeutic agent targeting the mucin 1 (MUC1) antigen. It may act by inducing the proliferation of interferon (IFN)-γ secreting MUC1-specific CD4+ T-cells and the generation of cytotoxic T lymphocytes capable of killing MUC1-expressing tumours. In the SPRINT (Stimuvax® [L-BLP25] in Rectal cancer In Neoadjuvant chemoradioTherapy) study, we will evaluate immune responses to L-BLP25 in patients with rectal cancer undergoing NCR. Study design SPRINT is a phase II, multi-centre, open-label study in which patients (n = 24/arm) with resectable stage II-III rectal cancer are randomized to NCR (capecitabine 825 mg/m2 twice-daily for 5–7 d/wk; 45–52 Gy in fractions of 1.8/2 Gy for ≥5 wk) alone, NCR + L-BLP25, or NCR + L-BLP25 + cyclophosphamide (CPA). L-BLP25 is administered as 8 consecutive weekly subcutaneous doses (930 µg) beginning on the first day of NCR, followed by a final injection 7–9 d before surgery. CPA (300 mg/m2; maximum 600 mg) is given as a single intravenous infusion 3 d before the first L-BLP25 dose. Surgery will be performed 6–8 wk after the end of NCR. The primary objective is to assess L-BLP25-induced changes in immune response profile. The intra-tumoural response will be evaluated based on analysis of tumour-infiltrating lymphocytes (especially CD8+ and CD45RO+). Peripheral antigen-specific response will be assessed by measuring IFN-γ secretion by peripheral blood mononuclear cells (ELIspot assay) in response to MUC1 and carcinoembryonic antigen (CEA) as a test for ‘antigen spreading’. Secondary objectives include the assessment of additional immunological changes and correlation of intratumoural, peripheral and skin delayed-type hypersensitivity responses. Safety and pathological anti-tumour responses in resection specimens will be evaluated. Disclosure A. Victor: Anja Victor is an employee of Merck KGaA Germany. S. Quaratino: Sonia Quaratino is an employee of Merck KGaA. All other authors have declared no conflicts of interest.

e14060 Background: The addition of anti Epidermal Growth Factor Receptor (EGFR) monoclonal antibodies (mabs) to preoperative chemoradiation in RC has produced low pathological complete response (pCR) rate. The postulated reasons for the poor outcome are: lack of patient selection, suboptimal treatment sequencing, and potential negative interaction between anti EGFR mabs and chemotherapy (CT) when used concurrently as radiosensitizers. Given these issues, we conducted a phase II trial of PAN in combination with preoperative RT in Kras wt RC. Methods: We studied several treatment sequences in murine models of Kras wt human colorectal cancer. Next, we used our preclinical results to design the treatment schedule of the phase II trial. PAN (6mg/kg/q2w) was combined with RT (45 Gy) to treat T3-4, Kras wt RC. Surgery was done 6-8 weeks after the end of RT. The primary endpoint, the pCR rate on surgical specimen, was assessed centrally (Simon: H0=5%, H1=17%, α=0.05, β=0.2). Results: In our murine models, the addition of anti EGFR mabs to RT significantly improved response when anti EGFR mabs were started during RT and extended after the end of RT (p50% pathological tumour regression (grade 3 Dworak) and 12 (70%) showed >2mm of tumour-free circumferential resection margin. Conclusions: Preclinical data confirm the need of optimal treatment sequencing to potentiate synergism between anti EGFR mabs and RT. However, the pCR rate in our phase II trial remains low despite adequate patient selection and optimized study design. Thus, we do not recommend the concurrent use of anti EGFR mabs and RT in the neoadjuvant treatment of RC.

Background Triple Negative Breast Cancers (TNBC) that represent 12% to 20% of total Breast Cancer (BC), have a worse outcome compared with other breast cancer subtypes. TNBC often show a deficiency in DNA double strand break repair mechanisms. This deficiency is generally related to inactivation of a repair enzymatic complex involving BRCA caused either by genetic mutations, epigenetic modifications, or by post-transcriptional regulations via miRNA. The identification of BC presenting a DNA double strand break repair deficiency could be useful to select patients that should get the best benefit from PARP inhibitors therapy. In this study, we have identified by Comparative Genomic Hybridization array (CGH-array) a recurrent gain characteristic of BRCA1 mutated tumours. Methods 135 formalin-fixed paraffin embedded (FFPE) tumours including TNBC (BRCA1 mutated and non-mutated), LumA, LumB and Her2-neu amplified BC were obtained from a local tumour collection. DNA was extracted and genomic copy-number alterations were analysed by CGH-array (Agilent, SuperPrint G3 Human CGH 8x60K Oligo Microarrays). Results In this study, we have identified by CGH-array a genomic region amplified in 90 % of the BRCA1 mutated tumours (29/32). A FISH assay on sections from FFPE samples was developed and confirmed the CGH profile. The identification of relevant genes whose expression is up-regulated within the recurrently gained region is underway. Conclusions The CGH profile and FISH assay developed in this study could allow the rapid identification of BRCAness breast tumours, improving the diagnostic performance and orienting the selection of the appropriate therapy. The up-regulated genes themselves might also represent potential therapeutic targets Acknowledgements This work was supported by Plan National Cancer, Belgium, action 29. Disclosure All authors have declared no conflicts of interest.

Investigation of methanol's surface chemistry on metals is a crucial step towards understanding the reactivity of this important chemical feedstock. Cu is a relevant metal for methanol synthesis and reforming, but due to the weak interaction of methanol with Cu, an atomic scale view of methanol's coverage-dependent ordering and self-assembly on Cu(111), the most abundant facet of most nanoparticles, has not yet been possible. Low and variable temperature scanning tunneling microscopy coupled with density functional theory reveal a coverage-dependent range of highly ordered structures stabilized by two hydrogen bonds per molecule. While extended chains that resemble the hydrogen-bonded zigzag structures reported for solid methanol are an efficient way to pack methanol at higher coverages, lower surface coverages yield isolated hexamer units. These hexamers form the same number of hydrogen bonds as the chains but appear to repel one another on the surface. Annealing treatments lead to the desorption of methanol with almost no decomposition. This data serves as a useful guide to both the preferred adsorption geometries and energies of a variety of methanol structures on Cu(111) surfaces as a function of surface coverage.

The aim of this study was to determine if patients with Borderline Personality Disorder (BPD) present higher emotional response than healthy controls in a laboratory setting. Fifty participants (35 patients with BPD and 15 healthy controls) underwent a negative emotion induction procedure (presentation of standardized unpleasant images). Subjective emotional responses were assessed by means of self-reported questionnaires while biological reactivity during the procedure was measured through levels of salivary cortisol (sCORT) and alpha-amylase (sAA). Patients with BPD exhibited significant lower cortisol levels and higher sAA levels compared to controls. Self-reported emotional reactivity did not give rise to differences between groups but participants with BPD did present higher levels of negative emotional intensity at baseline and during the entire procedure. The findings do not give support to the emotional hyperreactivity hypothesis in BPD. However, BPD patients presented heightened negative mood intensity at baseline, which should be considered a hallmark of the disorder. Further studies using more BPD-specific emotion inductions are needed to confirm the trends observed in this study El presente estudio pretende determinar si existe o no una mayor reactividad emocional en el Trastorno Límite de la Personalidad (TLP) en un contexto de laboratorio. Se realizó una inducción emocional negativa (presentación de imágenes estandarizadas con contenido negativo) a cincuenta participantes (35 pacientes con TLP y 15 controles sanos). Para evaluar la respuesta emocional subjetiva se utilizaron cuestionarios auto-informados; los niveles de cortisol (sCORT) y alfa-amilasa (sAA) salivares se utilizaron para medir la reactividad biológica al procedimiento. En el grupo de TLP, se observaron niveles de sCORT más bajos y niveles de sAA más elevados en comparación al grupo control. No se observaron diferencias significativas en relación a la reactividad emocional auto-informada, pero los pacientes con TLP reportaron mayor intensidad de emociones negativas a nivel basal así como también durante todo el procedimiento. Los resultados no apoyan la hipótesis de hiperreactividad emocional en el TLP. Sin embargo, los pacientes con TLP presentaron mayor intensidad de emociones negativas a nivel basal, característica que debería ser considerada como esencial en el trastorno. Futuros estudios deberán incorporar otros paradigmas de inducción emocional más específicos para TLP con el fin de confirmar las tendencias observadas en el presente estudio

This study assesses how the metastatic immune landscape is impacting the response to treatment and the outcome of colorectal cancer (CRC) patients.Complete curative resection of metastases (n = 441) was performed for two patient cohorts (n = 153). Immune densities were quantified in the center and invasive margin of all metastases. Immunoscore and T and B cell (TB) score were analyzed in relation to radiological and pathological responses and patient's disease-free (DFS) and overall survival (OS) using multivariable Cox proportional hazards models. All statistical tests were two-sided.The spatial distribution of immune cells within metastases was nonuniform. Patients, as well as metastases of the same patient, had variable immune infiltrates and response to therapy. A beneficial response was statistically significantly associated with increased immune densities. Among all metastases, Immunoscore (I) and TB score evaluated in the least immune-infiltrated metastases were the strongest predictors for DFS and OS (five-year follow-up, Immunoscore: I 3-4: DFS rate = 27.9%, 95% CI = 15.2 to 51.3; vs I 0-1-2: DFS rate = 12.3%, 95% CI = 4.9 to 30.6; HR = 0.45, 95% CI = 0.28 to 0.70, P = .02; I 3-4: OS rate = 64.6%, 95% CI = 46.6 to 89.6; vs I 0-1-2: OS rate = 32.5%, 95% CI = 17.2 to 61.4; HR = 0.32, 95% CI = 0.15 to 0.66, P = .001, C-index = 65.9%; five-year follow-up, TB score: TB 3-4: DFS rate = 25.7%, 95% CI = 14.2 to 46.6; vs TB 0-1-2: DFS rate = 5.0%, 95% CI = 0.8 to 32.4; HR = 0.36, 95% CI = 0.22 to 0.57, P.001; TB 3-4: OS rate = 63.7%, 95% CI = 46.4 to 87.5; vs TB 0-1-2: OS rate: 21.4%, 95% CI = 9.2 to 49.8; HR = 0.25, 95% CI = 0.12 to 0.51, P.001, C-index = 67.8%). High TB score and Immunoscore patients had a median survival of 70.5 months, while low patients survived only 25.1 to 38.3 months. Nonresponding patients with high-immune infiltrates had prolonged DFS (HR = 0.28, 95% CI = 0.15 to 0.52, P = .001) and OS (HR = 0.25, 95% CI = 0.1 to 0.62, P = .001). The immune parameters remained the only statistically significant prognostic factor associated with DFS and OS in multivariable analysis (P.001), while response to treatment was not.Response to treatment and prolonged survival of metastatic CRC patients were statistically significantly associated with high-immune densities quantified into the least immune-infiltrated metastasis.

We describe the weight's distribution in a sample of medical patients in hospital. We estimate the global prevalence and the presence between other different clinical variables.A prevalence cross-sectional study was carried out. We determine weight, stature and several clinical variables in 101 patients admit in the internal medicine department of Juan Ramón Jiménez hospital in Huelva. The patients were admitted from 6th to 7th of june in 2000. The Body Mass Index (BMI)or = 30 Kg/m2 was used to define the obesity.The prevalence of obesity was 32.2% [0.236-0.416]. In the study we find an association with female (prevalente rate -PR- 3.22), HTA (PR 4.72), dislipemia (RP 4.40) and hyperuricacemia (RP 4.28).The prevalence of obesity in our patients was between 23.41%, it was greater than others estimations in general people. We find association with women and classic cardiovascular risk factors.

Interleukin-6 (IL-6) is a major cytokine which is an essential mediator of cellular communication not only of the immune system but also of other physiological systems. Results with mice carrying a null mutation in the IL-6 gene (IL-6-/-) demonstrate that IL-6 is an essential factor for normal liver metallothionein (MT) response not only to inflammatory stimuli (endotoxin, turpentine) but also to a basically psychogenic stress model-immobilization stress. This indicates that this cytokine is an important component of the physiological response of the organism to stress in addition to its expected role during an immunological challenge. In contrast to the liver, in the brain the IL-6 deficiency only affected significantly MT-I+II response to inflammatory stimuli. Results with transgenic mice expressing IL-6 under the regulatory control of the glial fibrillary acidic protein gene promoter (GFAP-IL6) indicate that the MT-I+II isoforms are strongly up-regulated by IL-6. The fold induction in different brain areas correlates with the associated inflammatory response, and is comparable to that of the acute-phase response gene EB22/5, suggesting that MT-I+II could be considered acute-phase proteins in the brain. MT-III expression in the GFAP-IL6 mice is elevated in some areas (e.g. cerebellum) and decreased in others (e.g. occipital cortex), while that in IL-6-/- mice is slightly affected in a brain area-specific manner.

Heart failure (HF) and diabetes are common clinical conditions that may coexist. The main objective was to analyze the association of type-2 diabetes mellitus (T2DM) on prognosis in hospitalized patients with HF.We evaluated the association between T2DM with all-cause mortality and readmissions in the Spanish National Registry on Heart Failure-"Registro Nacional de Insuficiencia Cardiaca" (RICA). This is a multi-center and prospective cohort study that includes patients admitted for decompensated HF from 2008 to 2011. Study endpoints were all-cause mortality and hospital readmission. We determined the adjusted hazard ratio (HR) by a multivariable Cox regression model.A total of 1082 patients (mean age 77.6±8.5) were included of whom 490 (45.3%) had diabetes and 592 patients (54.7%) had preserved left ventricular ejection fraction (LVEF). During one-year follow-up, 287 patients died (151 patients with diabetes) and 383 patients were readmitted (197 patients with diabetes). After adjusting for baseline characteristics T2DM was significantly associated with all-cause mortality (HR 1.54; 95%CI 1.20-1.97, p=0.001) and readmissions (HR 1.46; 95%CI 1.18-1.80, p0.001). Age, dementia, peripheral vascular disease, NYHA class, renal insufficiency, hyponatremia and anemia were also independently associated with outcomes. There were no differences in mortality (p=0.415) and readmissions (p=0.514) according to preserved or reduced LVEF.T2DM is very common in patients hospitalized for HF. This condition is a strong and independent co-morbidity of all-cause mortality and readmission for both HF with preserved and reduced LVEF.

Background: Regorafenib, an oral multi-tyrosine kinase inhibitor that shares with sorafenib several targets on tumor angiogenesis, oncogenesis, and tumor microenvironment, was recently approved for patients (pts) with pretreated advanced colorectal cancer (aCRC). The drug improves the pts’ outcome, but with significant toxicities, underscoring the need to identify those who will not benefit. A previous study (SoMore trial) showed that early FDGPET-based metabolic response assessment (MRA) may adequately discriminate pts with chemorefractory aCRC unlikely to benefit from a sorafenib-capecitabine combination. RegARd-C aims to explore early MRA in pts treated with regorafenib as a clinical tool to spare pts from needless toxicity from a drug that gives them little or no benefit and as a translational tool to guide comprehensive genomic and epigenetic research on the determinants of drug resistance. Methods: RegARd-C’s (EUDRACT 2012-005655-16) is a multi-centric prospective study. Its primary objective is to identify in a population of pts with pretreated aCRC, those who will not benefit from regorafenib given at 160 mg/day, three weeks/4. Baseline PET is repeated at D14 of the first treatment course. MRA results are blinded for the investigators. Tumor tissues, optionally obtained from a PET-measurable lesion, and blood samples (at baseline; after the first chemotherapy course; and every two months) are collected. Overall survival (OS) is the primary endpoint and will be correlated with metabolic parameters, and genetic, epigenetic and molecular aberrations assessed from tumor biopsies and blood samples using gene expression and methylation profiling, RNA and exome sequencing. As the study is exploratory, no formal hypothesis was formulated. We arbitrarily decided to have a sample size of 105 evaluable pts with 70 pts as a derivation set and 35 pts as a validation set. Taking into account an expected 20-25% drop-out rate, between 124 and 140 pts will be accrued.This sample size is, however, sufficient to validate the hypothesis generated by the SoMore study, which found a prognostic impact of homogeneous metabolic response on OS, with an estimated HR of 0.59. RegARd-C has accrued 76 pts since August 2013. Clinical trial information: NCT01929616.