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In the last few decades, the use of ovarian stimulation with follicle stimulating hormone (FSH) or clomiphene citrate (CC) increases the percentage of multiple pregnancies. The presence of many follicles can lead to the risk of higher-order multiple pregnancy and ovarian high stimulation syndrome (OHSS). This is correlated to a significant increase in morbidity and mortality maternal and/or fetal rate. For this reason, sometime therapeutic measures are necessary to preserve woman’s health. We describe the case of a surgical abortion using a transcatheter arterial embolization (TAE), a minimally invasive, safe and effective procedure to preserve the patients.

Human BCL10 (hBCL10) protein is a signal transduction molecule originally identified because of its direct involvement in a subset of mucosa-associated lymphoid tissue (MALT) lymphomas, and later recognized as a crucial factor in regulating activation of NF-kB transcription factor following antigen receptor stimulation on lymphocytes. In this study, we characterized the NF-kB inducing activity of porcine BCL10 (pBCL10). pBCL10 oligimerizes, binds to components of the CARMA/ BCL10/MALT1 complex and forms cytoplasmic filaments. Functionally, in human cells pBCL10 is more effective in activating NF-kB compared to hBCL10, possibly due to the lack of carboxy-terminal inhibitory serine residues present in the human protein. Also, depletion experiments carried out through expression of short hairpin RNAs targeting hBCL10 indicate that pBcl10 can functionally replace the human protein and retains its higher NF-kB-inducing property in the absence of hBCL10. Our results contribute useful information on BCL10 protein in pigs, and may help the development of strategies based on the control of the immune response in pigs.

The three CARD-containing MAGUK (CARMA) proteins function as scaffolding molecules that regulate activation of the pro-inflammatory transcription factor NF-κB. Recently, mutations in CARMA2 have been linked to psoriasis susceptibility due to their acquired altered capacity to activate NF-κB. By means of two-hybrid screening with yeast, we identified RING finger protein 7 (RNF7) as an interactor of CARMA2. We present evidence that RNF7 functions as a negative regulator of the NF-κB-activating capacity of CARMA2. Mechanistically, RNF7 influences CARMA2 signaling by regulating the ubiquitination state of MALT1 and the NF-κB-regulatory molecule NEMO. Interestingly, CARMA2short (CARMA2sh) mutants associated with psoriasis susceptibility escape the negative control exerted by RNF7. In conclusion, our findings identify a new mechanism through which the ability of CARMA2 to activate NF-κB is regulated, which could have significant implications for our understanding of why mutations of this protein trigger human psoriasis.

Small molecule modulators of chromatin modifying enzymes became the focus of drug discovery efforts, because of their direct antitumor effects, such as apoptosis and cell-cycle arrest, in addition to their indirect immunomodulation properties (Peedicayil, 2012; West and Johnstone, 2014; Falkenberg and Johnstone, 2014). Recent studies have shown that inhibition of HDACs or POLA1 activity exerts anti-proliferative effects and also potentiates the immune response by activating type I Interferon (Medon et al., 2017; Starokadomskyy et al, 2016). To exploit a potential synergy, hybrid molecules targeting simultaneously POLA1 and HDACs, were designed. Here, we present in vitro and in vivo data concerning the antitumor activity of new dual POLA1-HDAC11 inhibitors. Among a library of screened molecules, MIR002 and MIR144 showed an antiproliferative activity at nanomolar concentrations on a panel of human tumor cell lines. In vitro functional assays revealed that these molecules potently inhibit POLA1 and specifically act on HDAC11 that recently emerged as regulator of several immune cells differentiation, pointing out its immunomodulatory role. Mechanistically, MIR002 and MIR144 induce acetylation of Histone H4 and alpha-Tubulin, canonical targets of HDACs, and p53-K382, as well. Activation of p53 leads to p21 increase and cell cycle arrest in the G1/S boundary. These compounds could be even more effective in treating human cancers when they are appropriately combined with other chemotherapeutics, such as Cisplatin. Indeed, the combination of MIR002 or MIR144 with Cisplatin, revealed a synergistic interaction (Combination Index < 1) on NCI-H460 and H460-R9A (ST1926-resistant sub-line) cell growth. The in vivo oral administration of MIR002 or MIR144 showed a potent antitumor activity, with a Tumor Growth Inhibition comprised between 60 and 100% in a series of human and murine solid and hematological tumors (human MM473 and MM432 Pleural Mesothelioma, H460 and H460R9A NSCLC, A2780-Dx and A2780-DDP ovarian carcinomas, NB-4 Acute Promyelocitic leukemia and Murine EL-4 Lymphoma) xenografted in nude and immunocompetent mice. The strongest antitumor effect of MIR144 was observed in immunocompetent mice rather than in nude mice suggesting that an immunomodulatory effect, along with the direct antiproliferative activity, contributes to tumor inhibition. Further analysis confirmed that MIR144 antitumor activity involves the induction/activation of several components of the host immune system, both at humoral and cellular levels. The large spectrum of antitumor activity, together with the high tolerability observed, open the possibility for their clinical investigation in different population of cancer patients. Citation Format: Claudio Pisano, Lucio Merlini, Sergio Penco, Raffaella Cincinelli, Nadine Darwiche, Mario B. Guglielmi, Ilaria La Porta, Maddalena Pizzulo, Federica Prosperi, Giacomo Signorino, Fabiana Colelli, Francesco Cardile, Laura Focareta, Alessandra Fucci, Egildo L. D'Andrea, Assunta Riccio, Sabrina Dallavalle. Host antitumor immune response activated by Dual POLA1-HDAC11 inhibitors endowed with a large spectrum of antitumor activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3096.

The molecular complexes formed by specific members of the family of CARMA proteins, the CARD domain-containing adapter molecule BCL10 and MALT1 (CBM complex) represent a central hub in regulating activation of the pleiotropic transcription factor NF-κB. Recently, missense mutations in CARMA2sh have been shown to cause psoriasis in a dominant manner and with high penetrancy. Here, we demonstrate that in human keratinocytes CARMA2sh plays an essential role in the signal transduction pathway that connects pathogen-associated molecular patterns recognition to NF-κB activation. We also find that the serine/threonine kinase ULK2 binds to and phosphorylates CARMA2sh, thereby inhibiting its capacity to activate NF-κB by promoting lysosomal degradation of BCL10, which is essential for CARMA2sh-mediated NF-κB signaling. Remarkably, CARMA2sh mutants associated with psoriasis escape ULK2 inhibition. Finally, we show that a peptide blocking CARD-mediated BCL10 interactions reduces the capacity of psoriasis-linked CARMA2sh mutants to activate NF-κB. Our work elucidates a fundamental signaling mechanism operating in human keratinocytes and opens to novel potential tools for the therapeutical treatment of human skin disorders.

Die Schweizer Automobilbranche befindet sich seit mehreren Jahren in einem kontinuierlichen Wachstum. Infolgedessen konnten die Absatzzahlen von neu immatrikulierten Personenfahrzeuge trotz eines turbulenten Jahres gegenüber dem Vorjahr steigen. Aus diesem Grund ist es in der heutigen Zeit für Unternehmungen von grosser Bedeutung, dass eine gute Kundenbindung geschaffen wird. Das Vertrauen einer Person gegenüber einer Automarke kann die Bindung stärken. Diese Arbeit beschreibt anhand bestehender Literatur aus welchen Elementen sich Vertrauen zusammensetzt. Hierbei wird zwischen den Komponenten affektives, kognitives und konatives Vertrauen unterschieden. Die Dimensionen Wohlwollen und Integrität sind dem affektiven Vertrauen zugeordnet, während die Kompetenz und Berechenbarkeit dem kognitiven Vertrauen zugesprochen werden. Zudem werden die wichtigsten Merkmale einer Marke eruiert. Im zweiten Teil dieser Arbeit werden in Bezug auf den Schweizer Automobilmarkt die neu immatrikulierten Personenfahrzeuge im Jahr 2015 genauer betrachtet. Anhand einer web-basierten Umfrage, die auf einem Modell zur Messung des Vertrauens basiert, wird untersucht, welcher Indikator den stärksten Einfluss auf das Gesamtvertrauen gegenüber einer Automarke hat. Zudem wird ebenfalls die einflussstärkste Dimension gegenüber dem Vertrauen bestimmt. Neben der Bestimmung der einflussstärksten Dimension des Vertrauens wird des Weiteren untersucht, ob bei gegebenem Vertrauen sich die Kaufabsicht, die Loyalität und die Weiterempfehlungsabsicht erhört. In einem letzten Schritt werden für die Automobilbranche geeignete Handlungsempfehlungen für den Auf- und Wiederaufbau von Vertrauen abgeleitet. Die Umfrage ist einerseits von zufällig ausgewählten Personen, und andererseits von Studenten der Zürcher Hochschule für Angewandte Wissenschaften aus dem Department School of Engineering beantwortet worden. Die Auswertungen erfolgten getrennt voneinander. Erst in einem zweiten Schritt sind beide Umfragen zusammengeführt und erneut ausgewertet worden. Bei der Analyse der Datensätze für alle drei Umfragen ist ersichtlich, dass alle Indikatoren und Dimensionen einen positiven Einfluss auf das Vertrauen haben. Die Stärke des Einflusses variiert sowohl je nach Indikator und Dimension, als auch je nach Umfrage. Insgesamt betrachtet, übt die Dimension der Kompetenz vom kognitiven Vertrauen, den stärksten Einfluss aus. Bei den Indikatoren hat die Produktkompetenz, welche besagt, dass die Automobilhersteller in Anbetracht deren Produkten und Leistungen kompetent sind, den stärksten Einfluss auf das Vertrauen. Ferner kam es zu dem Ergebnis, dass ein positiver Einfluss zwischen dem Handlungsentscheid der Befragten und dem Vertrauen gegenüber einer Automarke besteht.

The complexes formed by BCL10, MALT1 and specific members of the family of CARMA proteins (CBM complex), have recently focused much attention because they represent a central hub regulating activation of the transcription factor NF-κB following various cellular stimulations. In this manuscript, we report the functional characterization of a Danio rerio 241 amino acids polypeptide ortholog of the Caspase recruiting domain (CARD)-containing protein BCL10. Biochemical studies show that zebrafish Bcl10 (zBcl10) dimerizes and binds to components of the CBM complex. Fluorescence microscopy observations demonstrate that zBcl10 forms cytoplasmic filaments similar to that formed by human BCL10 (hBCL10). Functionally, in human cells zBcl10 is more effective in activating NF-κB compared to hBCL10, possibly due to the lack of carboxy-terminal inhibitory serine residues present in the human protein. Also, depletion experiments carried out through expression of short hairpin RNAs targeting hBCL10 indicate that zBcl10 can functionally replace the human protein. Finally, we show that the zebrafish cell line PAC2 is suitable to carry out reporter assays for monitoring the activation state of NF- kB transcription factor. In conclusion, this work shows that zebrafish may excellently serve as a model organism to study complex and intricate signal transduction pathways, such as those that control NF-κB activation.

We have developed stable cell lines expressing green fluorescent protein fusion proteins containing polyglutamine repeats of various lengths under tetracycline control. The expression of the expanded (43Q) repeat protein resulted in aggregate formation in a time-dependent fashion. The accumulation of aggregates did not induce apoptosis, although the survival of these cells was critically dependent on the presence of serum and growth factors. However, the expression of 43Q expanded protein strongly activated the ataxia telangiectasia mutated kinase/ATM and Rad3-related kinase (ATM/ATR)-dependent DNA damage response, as shown by selective phosphorylation of ATM substrates. This activation was dependent on 43 CAG protein expression, reversible and sensitive to caffeine and reducing agents. Similarly, we found phosphorylated ATM substrates in fibroblasts from Huntington's disease or SCA-2 patients. Oxidative stress induced accumulation of ATM/ATR phosphorylated protein in HD and SCA-2 patients, but not in normal controls. Furthermore, a significant phosphorylation of H2AX was shown by fibroblasts from patients. We conclude that polyglutamine induces ATM/ATR-dependent DNA damage response through accumulation of reactive oxygen species. ATM activation can be used to monitor the disease in vivo.

Aim: To analyze the prognostic value of maternal serum C-reactive protein (CRP) in predicting funisitis in patients with preterm premature rupture of membranes (pPROM). Methods: 66 patients (gestational age 24–33 weeks) hospitalized 1–12 h after pPROM were enrolled. White blood cell count (WBC), platelet count (PLT) and plasma concentration of CRP were assessed every 3 days. Histological evidence of chorioamnionitis and funisitis was obtained post-partum. Receiver operating characteristic (ROC) curves were employed to evaluate the role of maternal CRP in predicting funisitis. Results: Funisitis was found in 24 patients (36.3%); 42 patients (63.7%) without funisitis were considered as controls. PLT and WBC at admission and before delivery did not show significant differences and were not statistically different between the two groups. Patients with funisitis had significantly higher CRP levels both at admission to hospital and 24– 48 h before delivery. ROC curve analysis showed that CRP at admission (area under the curve: 0.671, p = 0.021) and before delivery (area under the curve: 0.737, p = 0.001) are predictive of funisitis. Conclusions: High maternal serum CRP levels (>20,000 µg/l) in pPROM patients at admission to hospital may be an early marker which indicates, with a good diagnostic performance, the presence of funisitis.

The aim of this study was to evaluate the correlations between the haematological parameter mean platelet volume and Doppler velocimetry parameters in order to improve clinical management in third trimester complicated pregnancies (pre-eclampsia, PE, and IUGR) affected by altered uterine resistances. Fifty-one patients were included in the abnormal uterine arteries Doppler velocimetry group (25 pregnancies were complicated by PE, 26 pregnancies were complicated by IUGR). Ninety-nine normotensive pregnant women taking no drugs for at least 2 weeks prior to testing and with no difference in gestational age at evaluation, with normal Doppler velocimetry profiles at routine screen, were used as controls. From all pregnant women, 20 mL of whole blood were obtained into citrate tubes after Doppler velocimetry evaluation and analysed for red blood cell counts (RBC), mean corpuscular volume (MCV), haemoglobin (HGB), haematocrit level (HCT), white blood cells count (WBC), platelet counts (PLT), mean platelets volume (MPV) and other biochemical parameters. From all blood parameters studied, MPV was significantly higher in women with altered uterine artery Doppler velocimetry compared with those with normal Doppler profiles (9.4 +/- 1.0 vs. 8.05 +/- 1.2 fL, P

Aims: To evaluate the effect of antenatal betamethasone and ritodrine in third trimester low risk singleton pregnancies by Doppler technology. Subjects and methods: Eighty-four third trimester pregnant women who received a full course of betamethasone and delivering uneventfully were recruited. The Doppler examination included the assessment of the pulsatility index (PI) of the umbilical artery (UA PI) and the middle cerebral artery (MCA PI) prior to treatment (baseline), and 48, 72 and 96 h after the second dose of betamethasone. Results: No significant difference was found in UA PI and UA/MCA values following betamethasone therapy. In contrast, MCA PI decreased significantly 48 h from the last injection of betamethasone in the whole study group (P