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Tire/road noise is a highly relevant topic for improving the comfort and experience of drivers and residents living in high-traffic areas. With the growing numbers of electric cars, the relevance of tire noise even increases since it is the dominant sound source in the middle-speed range. We investigate acoustic sources relevant to tire/road interactions. In doing so, we apply different sound source localization algorithms to measurement data acquired inside a large measurement trailer equipped with microphone arrays. The methods for sound source identification used are well-known beamforming-based algorithms and inverse schemes based on finite element or boundary element simulations. The latter schemes require the identification of the acoustic properties of the trailer in the stationary case. In this contribution, we present the characterization process and the results of the sound source localization in this stationary case.

Assessment of aspirin use and statistical analysis to assess an interaction of tumor MIR21 expression level and regular aspirin use after diagnosis in survival analysis

Purpose: Prostaglandin-endoperoxide synthase 2 (PTGS2, cyclooxygenase-2; a target of aspirin) produces inflammatory mediator prostaglandin E2 (PGE2), and contributes to colorectal neoplasia development. PTGS2-driven inflammatory responses can induce tumor expression of microRNA MIR21 (miR-21) that can increase local PGE2 level by downregulating PGE2-metabolizing enzymes. We hypothesized that the prognostic association of tumor MIR21 expression level in colorectal carcinoma might depend on inflammatory tumor microenvironment and be stronger in tumors expressing high-level PTGS2.Experimental Design: Utilizing 765 rectal and colon cancer specimens in the Nurses' Health Study and the Health Professionals Follow-up Study, we measured MIR21 expression by quantitative reverse transcription PCR, and PTGS2 expression by immunohistochemistry. Cox proportional hazards regression model was used to assess statistical interaction between MIR21 and PTGS2 in colorectal cancer–specific survival analysis, controlling for potential confounders including microsatellite instability, CpG island methylator phenotype, LINE-1 methylation level, and KRAS, BRAF, and PIK3CA mutations.Results: Tumor MIR21 expression level was associated with higher colorectal cancer–specific mortality (Ptrend = 0.029), and there was a statistically significant interaction between MIR21 and PTGS2 (Pinteraction = 0.0004). The association between MIR21 expression and colorectal cancer–specific mortality was statistically significant in PTGS2-high cancers (multivariable hazard ratio of the highest vs. lowest quartile of MIR21, 2.28; 95% confidence interval, 1.42–3.67; Ptrend = 0.0004) but not in PTGS2-absent/low cancers (Ptrend = 0.22).Conclusions:MIR21 expression level in colorectal carcinoma is associated with worse clinical outcome, and this association is stronger in carcinomas expressing high-level PTGS2, suggesting complex roles of immunity and inflammation in tumor progression. Clin Cancer Res; 22(15); 3841–8. ©2016 AACR.

Regular aspirin use after diagnosis and colorectal cancer mortality according to tumor MIR21 expression level among 579 patients with stage I to III colorectal cancer

The Diagnostic and Therapeutic Care of Abused Children and Adolescents In the following article, an overview of the current German health care situation concerning child abuse, neglect and maltreatment is provided. Interdisciplinary institutions of child protection are important initial contact points for families and professionals wishing to receive help. Furthermore, trauma clinics, victim support organizations and specialized counselling centers are indispensable to meet the complex needs of victims. Overall, the availability and accessibility of the support system in Germany is still insufficient and the number of unknown and untreated cases remains high. An improvement of the current situation should be of high governmental priority.

Background: Germline genetic factors central to immunity, such as human leukocyte antigen (HLA) variants, have been associated with several immune-related phenotypes such as response to immune checkpoint inhibitors. However, HLA and killer-cell immunoglobulin-like (KIR) gene combinations, which may modulate immune function, have not been studied in relation to T-cell density in cancer. Methods: This study was conducted within 3 well characterized epidemiologic studies that collected colorectal tumor tissue blocks (N=484). We profiled the in-situ T cell landscape of CRC using digital imaging, machine learning, and a customized 9-plex multiplexed immunofluorescence panel with antibodies directed against CD3, CD4, CD8, CD45RA, CD45RO, FOXP3, and MKI67. HLA and KIR variants were imputed from genome-wide array datasets through SNP2HLA and KIR*IMP methods. We used multivariable ordinal logistic regression to estimate odds ratios (ORs) and 95% confidence intervals (CIs) for the association between HLA-KIR activation/inhibition ligand combinations with quartile of T cell densities in CRC adjusting for age, sex, GWAS panel, and the first two principal components of ancestry. Results: Presence of KIR2DL2+HLAC1+ and KIR2DS2+HLAC1+ combinations were associated with lower odds of greater CD3+CD8+ T-cell density quartile, however these findings were not statistically significant [OR = 0.73, 95% CI (0.52, 1.02), p-value = 0.067; OR = 0.74 95% CI (0.53, 1.04), p-value= 0.082, respectively]. There was no association between HLA-KIR combinations and CD3+CD4+ T-cell density quartile. Further results will examine HLA and KIR genes individually, as well as additional combination variables and more specific T-cell density subsets. Conclusions: Further investigation is needed to determine if germline genetics related to immune profile plays a role in T-cell densities in CRC. Citation Format: Claire Elizabeth Thomas, Jeroen Huyghe, Tomotaka Ugai, Hang Yin, Yasutoshi Takashima, Daniel D. Buchanan, Conghui Qu, Li Hsu, Andressa Dias Costa, Stephen Gallinger, Robert Grant, Sushma Thomas, Shuji Ogino, Amanda I. Phipps, Jonathan Nowak, Ulrike Peters. Association between HLA-KIR allele interaction combinations and density of T-cell subsets in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3014.

Background: Despite heightened interest in early-onset CRC, little is known about the tumor molecular, immune, and microbial characteristics of early-onset CRC. It is also unclear whether CRCs diagnosed at or shortly after age 50 are similar to early-onset CRC. We hypothesized that tumor molecular, immune, and microbial characteristics in CRC tissue might show differential heterogeneity patterns between three age groups ( Methods: We examined 3,395 CRC cases with available tissue data, including 660 early-onset and 243 intermediate-onset cases in the Nurses' Health Study, Health Professionals Follow-up Study, and Ontario Familial Colon Cancer Registry. We profiled the in situ T-cell landscape of 959 cases using digital imaging, machine learning, and a customized 9-plex multiplexed immunofluoresence panel with antibodies directed against CD3, CD4, CD8, CD45RA, CD45RO, FOXP3, and MKI67 (Ki-67). Using chi-squared test or Spearman’s correlation test, we assessed differences in tumor characteristics including microsatellite instability, CpG island methylator phenotype (CIMP), KRAS, and BRAF mutations, LINE-1 methylation levels, pks+ E. coli and Fusobacterium nucleatum positivity, histopathologic lymphocytic reaction, and T-cell densities between early-onset, intermediate-onset, and later-onset cases. Results: Compared to later-onset CRC (21%), early-onset (4.8%) and intermediate-onset CRCs (5.1%) exhibited a lower prevalence of CIMP-high status (P Conclusions: Compared to later-onset CRC, early-onset and intermediate-onset CRCs tended to have aggressive tumor phenotypes such as LINE-1 hypomethylation, lower lymphocytic immune reaction, and a higher density of MKI67+ immune cells. These findings highlight the importance of the tumor microenvironment in the etiology of early-onset and intermediate-onset CRCs. Citation Format: Tomotaka Ugai, Yasutoshi Takashima, Andressa Dias Costa, Daniel Buchanan, Jeroen Huyghe, Li Hsu, Conghui Qu, Claire Thomas, Steve Gallinger, Robert Grant, Ulrike Peters, Amanda I. Phipps, Jonathan Nowak, Shuji Ogino. Molecular, immune, and microbial profiles of early-onset, intermediate-onset, and later-onset CRCs. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5760.

Background. Prior studies have demonstrated that the overall density of T cells in colorectal tumors is favorably associated with colorectal cancer (CRC) survival; however, few studies have considered the potentially distinct roles of heterogeneous T cell subsets in different tissue regions in relation to CRC outcomes. Methods. We used data and colorectal tumor specimens from a subset of participants in three studies (total N=878). We profiled the in situ T cell landscape of CRC using digital imaging, machine learning, and a customized 9-plex multiplexed immunofluoresence panel with antibodies directed against CD3, CD4, CD8, CD45RA, CD45RO, FOXP3, and MKI67. This allowed us to determine the density of T cell subsets defined according to marker co-localization and according to tissue region (i.e., epithelial, stromal). We used Cox proportional hazards models to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) for the associations of T cell subset densities in CRC with disease-specific survival, adjusting for age, sex, tumor site, microsatellite instability status, CpG island methylator phenotype status, status of somatic mutations in BRAF and KRAS, stage, and study. Results. Overall, the densities of CD3+CD4+ helper T cells and CD3+CD8+ cytotoxic T cells in CRC were favorably associated with CRC-specific survival. However, in analyses stratified by tissue region, the association of CD3+CD4+ helper T cell density with survival was limited to density in stromal regions (HR for highest vs. lowest density quartile=0.40, 95% CI: 0.26-0.63, ptrend Conclusions. The density of T cells in CRC tissue is significantly associated with CRC survival, independent of stage at diagnosis and other evaluated tumor markers; however, the T cell subsets that are associated with CRC outcomes differ according to tissue region. These findings highlight the differing roles of distinct T cell subsets in CRC with regard to cancer outcomes, and support the need for additional work identifying factors that contribute to T cell response profiles in CRC. Citation Format: Amanda I. Phipps, Jonathan Nowak, Yasutoshi Takashima, Daniel Buchanan, Andressa Dias Costa, Steven Gallinger, Jeroen Huyghe, Li Hsu, Conghui Qu, Claire Thomas, Sushma Thomas, Tomotaka Ugai, Ulrike Peters, Shuji Ogino. Density of T cell subsets in colorectal cancer in relation to disease-specific survival [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3019.

794 Background: Tumor genomic testing (GT) has increased diagnostic accuracy and treatment options for patients (pts) with cancer. Dana-Farber Cancer Institute (DFCI) has made GT accessible as an institute-supported research effort for >10 yrs. We estimate 50% standard therapies and 15-35% clinical trials in Gastrointestinal Cancer Clinic (GCC) require GT to determine eligibility. Pts in GCC with certain cancers are eligible for GT as a clinical test – these include metastatic/locally advanced colorectal, gastric, pancreatic, or biliary cancers. Clinical testing requires CLIA lab certification and insurance reimbursement; research does not. Herein we ID gaps in our GT database. Methods: We reviewed data on GT uptake in GCC between 4/2015 - 6/2022. 20,096 pts were captured by the GT tracking system. Data included: testing ordered and completed (proportion, type, time to receiving tissue for testing [TR], time to testing completion [TC]). Demographic data is not captured in the tracking system; matching unique patient identifiers with electronic health record is pending. Results: Most pts received GT (57.6%); 12% were not eligible; 30.4% declined consent. Most testing was completed (67.6%), but 21.3% of tests failed (45.5% of these from insufficient tissue). Research testing (71%) comprised most tests, but clinical tests were completed faster (median 34 days research vs 20 days clinical). Ampullary (91%), anal (90%), colon (90%) had highest completion rates; pancreatic (59%), hepatocellular carcinoma (56%) had lowest (from insufficient viable tumor in submitted specimens). Conclusions: GCC has a robust recruitment program that has yielded high GT uptake. Given the frequency that GT is used for treatment and trials, building a demographically representative dataset is crucial, especially for pts with largest burden of morbidity and mortality from cancer. We ID'd data gaps in the GT tracking system, which lacks demographics and reason for not testing. Demographic data is available in the electronic health record but does not speak with the GT tracking system so this analysis is not routinely done. Ability to visualize this data is important to ensure equitable GT uptake. Future efforts will focus on improving rates of consent in genomics databases and cancer clinical trials. Genomic testing at Dana-Farber Cancer Institute Gastrointestinal Cancer Center, 4/2015 – 6/2022.[Table: see text]

Pancreatic ductal adenocarcinoma (PDA) is the third most common cause of cancer death in the US with a 11% 5-year survival rate. A key contributor of this dismal prognosis is early and frequent metastatic spread with limited treatment options for patients. Metastasis in PDA is facilitated by a robust fibroinflammatory response in the primary tumor site, a large portion of which is composed of infiltrating immune cells, and survival during transit to establish in the secondary site. In the primary tumor reactive stroma, we found increased eosinophils in both human and mouse PDA compared to the normal pancreas. Eosinophils are an innate immune cell best known for their ability to control parasite responses and have been implicated in either pro- or anti-cancer roles in various solid tumors. We used the ΔdblGata mice, which lack eosinophils, to study the role of eosinophils during metastatic spread using a resectable model of PDA metastasis. This model of metastasis was developed to facilitate consistent seeding of lung metastasis in approximately 50 percent of mice from a transient subcutaneous tumor which undergoes subsequent surgical removal and harvest to evaluate metastasis at 4 weeks. We found that mice lacking eosinophils have increased metastasis to the lung in our resectable model of PDA metastasis but no difference in establishment of IV injected PDA tumor cells, suggesting eosinophil control at the primary tumor site. Single cell sequencing of the subcutaneous tumors finds differences in the angiogenic subtypes, tip and stalk cells, in mice lacking eosinophils. Sequencing of eosinophil populations in humans and mice find an increase in angiogenic signaling suggesting a regulatory role in tumor cell access to the circulation. We conclude that eosinophils in PDA inhibit the metastatic process through control of angiogenic signaling. Citation Format: Megan T. Hoffman, Patrick J. Lenehan, Jennifer Wang, Lestat Ali, Li Qiang, Amiko Uchida, Sara Vayrynen, Andressa Dias Costa, Michael Dougan, Max Heckler, Jonathan Nowak, Brian W. Wolpin, Stephanie K. Dougan. Eosinophils alter metastatic spread in pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer; 2022 Sep 13-16; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2022;82(22 Suppl):Abstract nr B018.

In preclinical work, the inflammatory cytokine IL-1β was shown to be upregulated in pancreatic cancer tumors and to contribute to activation of pancreatic stellate cells and immunosuppression (Das et al 2020). We conducted an open-label multicenter Phase Ib study evaluating gemcitabine, nab-paclitaxel, canakinumab (ACZ885), a high-affinity human anti-interleukin-1β (IL-1β) mAb, and spartalizumab (PDR001), a PD-1 mAb. Eligible subjects had previously untreated metastatic PDA and RECIST measurable disease. The primary objective was to confirm recommended phase II dose by evaluating the incidence of dose limiting toxicities (DLTs) in the first 56 days of dosing in at least 6 evaluable subjects utilizing a Bayesian logistic regression model. Subjects underwent baseline and on-study tissue and blood collection for correlative translational research. Results: 10 subjects were enrolled between Nov 2020 and Mar 2021. At the primary data cut off of May 23, 2021, 6 subjects were evaluable for DLT. There were no DLTs, the recommended Phase II dose was established as: gemcitabine (1000mg/m2 IV) day 1,8,15; nab-paclitaxel (125mg/m2 IV) day 1,8,15; canakinumab (250mg SC) day 1, spartalizumab (400mg IV) day 1; of each 28 day cycle. At the time of an updated database extraction on June 1, 2022, 2 subjects remain on study. Adverse events were consistent with those typically seen with chemotherapy. The most common Grade 3/4 AEs were neutropenia (60%) and anemia (50%), with no fatal AEs. One patient discontinued spartalizumab due to grade 3 pneumonitis. In preliminary efficacy analysis (n=10), there are 3 confirmed PRs, 5 subjects with stable disease, 2 subjects with progression as best response. Individual site data estimates that the 12 month OS rate is 60%; updated RR, PFS and OS data will be reported. Activation of CD8 T cells in peripheral blood and increased serum levels of IFN-induced chemokines CXCL9/10 were observed in both responder and non-responder patients. Using an in vitro suppression assay, we showed that baseline serum from responders could induce myeloid derived suppressor cells, an effect that was abrogated with treatment. Single cell transcriptional profiling, multiplex immunofluorescence and spatial transcriptomics also revealed treatment-dependent shifts in T cell activation state and myeloid cells in the tumors of patients experiencing clinical response. Conclusions: PanCAN-SR1 established the Phase II dose of canakinumab and spartalizumab with chemotherapy in first line metastatic PDA, based upon favorable benefit-risk assessment. Based on our comprehensive analysis of 10 patients treated with combination therapy including IL-1b blockade, we hypothesize that the definitive role of anti-IL-1b in human patients with pancreatic cancer is to reduce systemic immune suppression and to reduce immunosuppressive myeloid cell activation in the tumor. The clinical utility of targeting IL-1β in pancreatic cancer is being evaluated in a randomized Phase II/III study through the Precision Promise clinical trial network. NCT04581343. Citation Format: Paul E. Oberstein, Osama Rahma, Nina Beri, Amy Stoll-D'Astice, Emily A. Kawaler, Igor Dolgalev, Gregor Werba, Victoire Cardo-Ruffino, Naïma Böllenrücher, Andressa Dias Costa, Saloney Nazeer, Matthew Squires, Jonathan Nowak, Dafna Bar-Sagi, Brian Wolpin, Diane M Simeone, Stephanie K Dougan. Primary results of PanCAN-SR1, a phase 1b study evaluating Gemcitabine, nab-Paclitaxel, Canakinumab, and Spartalizumab to target IL-1β and PD-1 in metastatic pancreatic cancer with correlative tissue and blood biomarker analysis [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer; 2022 Sep 13-16; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2022;82(22 Suppl):Abstract nr PR005.

Zusammenfassung Im vorliegenden Artikel wird eine Übersicht zu den Versorgungsmöglichkeiten von Kindern und Jugendlichen mit Misshandlungs-, Missbrauchs- und Vernachlässigungserfahrungen in Deutschland gegeben. Interdisziplinäre und transprofessionelle Institutionen wie z. B. eine Kinderschutzambulanz oder ein Childhood-Haus spielen in der Abklärung eine zentrale Rolle. Zudem sind aufgrund des meist komplexen Behandlungs- und Beratungsbedarfs der Betroffenen Traumaambulanzen, Opferhilfeorganisationen und Fachberatungsstellen unverzichtbar. Jedoch ist die flächendeckende Verfügbarkeit solcher Angebote bislang in Deutschland nur unzureichend, weshalb viele Fälle weiterhin unerkannt und unbehandelt bleiben. An dieser Stelle sollte dringend Verantwortung durch Kommunen und Länder übernommen werden.

Previous research has demonstrated that loss of sleep has a negative impact on both emotional and cognitive functioning. We examined whether subjectively reported natural sleep loss is associated with the interplay between emotion and cognition, as was probed by brain activity in response to emotional distraction during a working memory task. Forty-six healthy male adults reported their typical weekly sleep pattern using the Munich Chronotype Questionnaire (MCTQ), while recent sleep loss was enquired using a sleep diary in the 7 days preceding scanning. Participants performed a delayed match-to-sample task with negative and neutral distracters during the delay period inside the MRI scanner. Activity differences between negative and neutral distracters were associated to both sleep loss measures across participants. The amount of typically encountered sleep loss indicated by the MCTQ, but not sleep diary, was negatively associated with activity in the rostral anterior cingulate cortex and dorsomedial prefrontal cortex during emotionally negative compared to neutral distraction (p < 0.025, whole brain corrected). Participants showed less distracter-related activity in the ACC and dorsomedial PFC with increasing sleep loss, which, in the long run, might contribute to less adaptive emotional processing, and therefore a greater vulnerability to develop affective disorders.

Introduction: One of the factors that limit the efficacy of the drugs, especially in primary brain tumors, is the permeability of the blood-brain barrier (BBB). The low-density lipoprotein receptor (LDL-R) expressed at the BBB mediates the transport of endogenous ligands through the BBB. We developed a new radiolabeled peptide targeting both the human and murine LDLR and able i) to cross the BBB and ii) to target tumors such as glioblastoma that express high levels of the LDLR. The objective of this study was to determine the LDLR targeting properties, the pharmacokinetics of 68Ga radiolabeled RMX-VH in a glioblastoma model that expresses the human LDLR (hLDLR). Methods: The nonclinical studies of 68Ga-RMX-VH were completed in U87MG, A172, U373 glioblastoma cancer cell lines and xenografts mice models. The studies determined in vivo time-dependent accumulation of this agent, in vitro dose-depended cellular uptake, and cellular competition studies. We compared the tumor-specific accumulation of 68Ga-RMX-VH and normal organ distribution in female and male athymic nude mice. Radiotracer, RMX-VH (10-30ug) was labeled with isotope-Ga68 (10-25mCi, ITM GmBH). U87MG and A172- derived xenografts were generated in athymic nude mice (10 weeks) and PET/CT images were acquired using G4 PET/Xray camera (Sofie Biosciences) at 1h, 2h, 3h, and 4h post-injection. The followed up biodistribution studies were done at 30 min, 1h, 2h, and 3 hrs. post-injection. The organs and tumor were collected, weighed, and the tissue radioactivity was measured with Wizard2 Gamma Counter (Perkin-Elmer, Waltham, MA). The percentage of injected dose per gram of tissue (%ID/g) was calculated and decay-corrected. Results: Our studies confirmed the in vitro and in vivo selectivity and specificity of 68Ga-RMX-VH toward LDLR-positive tumors. 68Ga-RMX-VH is a small peptide (MW: 1432.7 g/mol) and renal excretion was expected as a route of agent elimination. The tumor-specific uptake of radiotracer in U87MG xenografts was 1.8%ID/g at 1h and remained unchanged at 3h post-injection. The kidney retention of the agent reached 12.2%ID/g and decreased to 10.2%ID/g. The accumulation of radiotracer in the liver, ovaries, and intestine correlated well with the known normal expression of LDLR. The tumor-to-muscle ratio was 5.97 at 1h; increased to 26.1 and 22.39 at 2h and 3h. The elimination t1/2 of radiotracer was only 18.6 min, with a clearance CL of 502 ml/min. Adsorption from the site of administration is rapid as the Cmax was 5 min. The tumor-specific uptake and normal organ distribution of 68Ga-RMX-VH were lower in male mice than in females. This correlated with differences in lipid and lipoproteins metabolism in males and females. Conclusions: RMX-VH showed favorable hLDLR targeting properties in vitro and in vivo in glioblastoma mice models. Our results suggest that hLDLR may serve as a target for imaging for glioblastoma. The first-in-human exploratory IND study of Ga68-RMX-VH will be initiated in Q2 of 2021. Citation Format: Izabela Tworowska, Leo Garcia Flores II, Xuewei Qu, Cédric Malicet, Nilesh Wagh, David Ranganathan, Jonathan Nowak, Pascaline Lecorche, Jamal Temsamani, Ebrahim S. Delpassand. Radionuclide imaging of low-density-lipoprotein receptor (LDLR)-overexpressing glioblastoma: A preclinical study of Gallium-68 RMX-VH [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2021 Oct 7-10. Philadelphia (PA): AACR; Mol Cancer Ther 2021;20(12 Suppl):Abstract nr P099.

Enduring sleep loss is a risk factor for a variety of both somatic and mental health issues. When subjected to sleep loss, the brain becomes vulnerable to critical alterations in cognitive and emotional processing. In our study, we examined the effect of psychosocial stress on amygdala resting-state functional connectivity in participants with cumulative sleep loss calculated across the seven days preceding scanning. For this purpose, forty-five healthy male participants completed a one-week sleep diary and underwent resting-state scans before and after taking part in the ScanSTRESS paradigm, which allows social stress induction during functional magnetic resonance imaging. Sleep loss was negatively associated with seed-based functional connectivity of the left amygdala with the medial prefrontal cortex, dorsal anterior cingulate cortex, anterior insula, posterior cingulate cortex, and dorsolateral prefrontal cortex. That is, participants with higher amounts of sleep loss showed reduced left amygdala connectivity after social stress induction to cortical regions encompassing main nodes of the brain's default mode network and salience network. Our results shed more light on how brain functional connectivity may shape the brain's stress response in the context of naturally occurring sleep loss, revealing a potential neural mechanism for increased vulnerability to stress-related psychopathology.

International audience; Using a bioinformatic approach, we identified a TP53INP1-related gene encoding a protein with 30% identity with tumor protein 53-induced nuclear protein 1 (TP53INP1), which was named TP53INP2. TP53INP1 and TP53INP2 sequences were found in several species ranging from Homo sapiens to Drosophila melanogaster, but orthologues were found neither in earlier eukaryotes nor in prokaryotes. To gain insight into the function of the TP53INP2 protein, we carried out a yeast two-hybrid screening that showed that TP53INP2 binds to the LC3-related proteins GABARAP and GABARAP-like2, and then we demonstrated by coimmunoprecipitation that TP53INP2 interacts with these proteins, as well as with LC3 and with the autophagosome transmembrane protein VMP1. TP53INP2 translocates from the nucleus to the autophagosome structures after activation of autophagy by rapamycin or starvation. Also, we showed that TP53INP2 expression is necessary for autophagosome development because its small interfering RNA-mediated knockdown strongly decreases sensitivity of mammalian cells to autophagy. Finally, we found that interactions between TP53INP2 and LC3 or the LC3-related proteins GABARAP and GABARAP-like2 require autophagy and are modulated by wortmannin as judged by bioluminescence resonance energy transfer assays. We suggest that TP53INP2 is a scaffold protein that recruits LC3 and/or LC3-related proteins to the autophagosome membrane by interacting with the transmembrane protein VMP1. It is concluded that TP53INP2 is a novel gene involved in the autophagy of mammalian cells.

Shaping economic development to be environmentally compatible is becoming increasingly urgent. The wind turbine industry has been successfully developed as a domestic industry in both China and India. Important success factors here are policies to facilitate learning and integrated environmental, technological and industrial policy schemes. There are clear differences between the countries, e.g. with regard to the importance of electric utilities, the international focus of the main players and the level of industrial policies to support domestic suppliers. There is not just ‘one successful way’ of developing a domestic wind turbine industry: India has taken a more market driven approach compared to China, where the development is more government induced and is taking place even more rapidly.

We identified a TP53INP1-related gene, which was named TP53INP2, using a bioinformatic approach. TP53INP1 and TP53INP2 sequences were found in higher eukaryotes but not in lower eukaryotes nor in prokaryotes. To gain insight into the function of the TP53INP2 protein, we carried out a number of protein-protein interaction approaches which showed that TP53INP2 binds to the Atg8-like proteins, as well as with the autophagosome trans-membrane protein VMP1. TP53INP2 translocates from the nucleus to autophagosomes after activation of autophagy. We also showed that TP53INP2 expression is necessary for autophagosome development since its siRNA-mediated knock-down strongly decreases sensitivity to induced-autophagy. We suggest that TP53INP2 is a scaffold protein that recruits Atg8-like proteins to the autophagosome membrane by interacting with the trans-membrane protein VMP1.

14 pages, 8 figures., Autophagy can promote cell survival or cell death, but the molecular basis underlying its dual role in cancer remains obscure. Here we demonstrate that Δ9-tetrahydrocannabinol (THC), the main active component of marijuana, induces human glioma cell death through stimulation of autophagy. Our data indicate that THC induced ceramide accumulation and eukaryotic translation initiation factor 2α (eIF2α) phosphorylation and thereby activated an ER stress response that promoted autophagy via tribbles homolog 3–dependent (TRB3-dependent) inhibition of the Akt/mammalian target of rapamycin complex 1 (mTORC1) axis. We also showed that autophagy is upstream of apoptosis in cannabinoid-induced human and mouse cancer cell death and that activation of this pathway was necessary for the antitumor action of cannabinoids in vivo. These findings describe a mechanism by which THC can promote the autophagic death of human and mouse cancer cells and provide evidence that cannabinoid administration may be an effective therapeutic strategy for targeting human cancers., This work was supported by grants from the Spanish Ministry of Education and Science (MEC) (HF2005/0021, to G. Velasco; SAF2006/00918, to M. Guzmán; and BFU2006-00508, to P. Boya), Santander-Complutense PR34/07-15856, to G. Velasco), Comunidad de Madrid (S-SAL/0261/2006, to M. Guzmán), and La Ligue contre le Cancer and Canceropole PACA (to J.L. Iovanna). M. Salazar was the recipient of a fellowship from the MEC. A. Carracedo was the recipient of fellowships from Gobierno Vasco, the Federation of European Biochemical Societies, and the European Molecular Biology Organization. M. Lorente and P. Boya have a Juan de la Cierva and a Ramón y Cajal contract from the MEC, respectively. S. Hernández-Tiedra has a technician contract from the Spanish Ministry of Education and the Fondo Social Europeo.

BACKGROUND: Pancreatic cancer cells generate metastases because they can survive the stress imposed by the new environment of the host tissue. To mimic this process, pancreatic cancer cells which are not stressed in standard culture conditions are injected into nude mice. Because they develop xenografts, they should have developed adequate stress response. Characterizing that response might provide new strategies to interfere with pancreatic cancer metastasis. METHODOLOGY/PRINCIPAL FINDINGS: In the human pancreatic cancer cell lines Panc-1, Mia-PaCa2, Capan-1, Capan-2 and BxPC3, we used Affymetrix DNA microarrays to compare the expressions of 22.000 genes in vitro and in the corresponding xenografts. We identified 228 genes overexpressed in xenografts and characterized the implication of one of them, WSB1, in the control of apoptosis and cell proliferation. WSB1 generates 3 alternatively spliced transcripts encoding distinct protein isoforms. In xenografts and in human pancreatic tumors, global expression of WSB1 mRNA is modestly increased whereas isoform 3 is strongly overexpressed and isoforms 1 and 2 are down-regulated. Treating Mia-PaCa2 cells with stress-inducing agents induced similar changes. Whereas retrovirus-forced expression of WSB1 isoforms 1 and 2 promoted cell growth and sensitized the cells to gemcitabine- and doxorubicin-induced apoptosis, WSB1 isoform 3 expression reduced cell proliferation and enhanced resistance to apoptosis, showing that stress-induced modulation of WSB1 alternative splicing increases resistance to apoptosis of pancreatic cancer cells. CONCLUSIONS/SIGNIFICANCE: Data on WSB1 regulation support the hypothesis that activation of stress-response mechanisms helps cancer cells establishing metastases and suggest relevance to cancer development of other genes overexpressed in xenografts.

Autophagy is a degradation process of cytoplasmic cellular constituents, which serves as a survival mechanism in starving cells, and it is characterized by sequestration of bulk cytoplasm and organelles in double-membrane vesicles called autophagosomes. Autophagy has been linked to a variety of pathological processes such as neurodegenerative diseases and tumorigenesis, which highlights its biological and medical importance. We have previously characterized the vacuole membrane protein 1 (VMP1) gene, which is highly activated in acute pancreatitis, a disease associated with morphological changes resembling autophagy. Here we show that VMP1 expression triggers autophagy in mammalian cells. VMP1 expression induces the formation of ultrastructural features of autophagy and recruitment of the microtubule-associated protein 1 light-chain 3 (LC3), which is inhibited after treatment with the autophagy inhibitor 3-methiladenine. VMP1 is induced by starvation and rapamycin treatments. Its expression is necessary for autophagy, because VMP1 small interfering RNA inhibits autophagosome formation under both autophagic stimuli. VMP1 is a transmembrane protein that co-localizes with LC3, a marker of the autophagosomes. It interacts with Beclin 1, a mammalian autophagy initiator, through the VMP1-Atg domain, which is essential for autophagosome formation. VMP1 endogenous expression co-localizes with LC3 in pancreas tissue undergoing pancreatitis-induced autophagy. Finally, VMP1 stable expression targeted to pancreas acinar cell in transgenic mice induces autophagosome formation. Our results identify VMP1 as a novel autophagy-related membrane protein involved in the initial steps of the mammalian cell autophagic process.

TP53INP1 is an alternatively spliced gene encoding two nuclear protein isoforms (TP53INP1alpha and TP53INP1beta), whose transcription is activated by p53. When overexpressed, both isoforms induce cell cycle arrest in G1 and enhance p53-mediated apoptosis. TP53INP1s also interact with the p53 gene and regulate p53 transcriptional activity. We report here that TP53INP1 expression is induced during experimental acute pancreatitis in p53-/- mice and in cisplatin-treated p53-/- mouse embryo fibroblasts (MEFs). We demonstrate that ectopic expression of p73, a p53 homologue, leads to TP53INP1 induction in p53-deficient cells. In turn, TP53INP1s alters the transactivation capacity of p73 on several p53-target genes, including TP53INP1 itself, demonstrating a functional association between p73 and TP53INP1s. Also, when overexpressed in p53-deficient cells, TP53INP1s inhibit cell growth and promote cell death as assessed by cell cycle analysis and colony formation assays. Finally, we show that TP53INP1s potentiate the capacity of p73 to inhibit cell growth, that effect being prevented when the p53 mutant R175H is expressed or when p73 expression is blocked by a siRNA. These results suggest that TP53INP1s are functionally associated with p73 to regulate cell cycle progression and apoptosis, independently from p53.