A Hippo and Fibroblast Growth Factor Receptor Autocrine Pathway in Cholangiocarcinoma

Herein, we have identified cross-talk between the Hippo and fibroblast growth factor receptor (FGFR) oncogenic signaling pathways in cholangiocarcinoma (CCA). Yes-associated protein (YAP) nuclear localization and up-regulation of canonical target genes was observed in CCA cell lines and a patient-derived xenograft (PDX). Expression of FGFR1, -2, and -4 was identified in human CCA cell lines, driven, in part, by YAP coactivation of TBX5. In turn, FGFR signaling in a cell line with minimal basal YAP expression induced its cellular protein expression and nuclear localization. Treatment of YAP-positive CCA cell lines with BGJ398, a pan-FGFR inhibitor, resulted in a decrease in YAP activation. FGFR activation of YAP appears to be driven largely by FGF5 activation of FGFR2, as siRNA silencing of this ligand or receptor, respectively, inhibited YAP nuclear localization. BGJ398 treatment of YAP-expressing cells induced cell death due to Mcl-1 depletion. In a YAP-associated mouse model of CCA, expression of FGFR 1, 2, and 4 was also significantly increased. Accordingly, BGJ398 treatment was tumor-suppressive in this model and in a YAP-positive PDX model. These preclinical data suggest not only that the YAP and Hippo signaling pathways culminate in an Mcl-1-regulated tumor survival pathway but also that nuclear YAP expression may be a biomarker to employ in FGFR-directed therapy.     

Synthesis and evaluation of modified chalcone based p53 stabilizing agents

Tumor suppressor protein p53 induces cell cycle arrest and apoptotic cell death in response to various cellular stresses thereby preventing cancer development. Activation and stabilization of p53 through small organic molecules is, therefore, an attractive approach for the treatment of cancers retaining wild-type p53. In this context, a series of nineteen chalcones with various substitution patterns of functional groups including chloro, fluoro, methoxy, nitro, benzyloxy, 4-methyl benzyloxy was prepared using Claisen-Schmidt condensation. The compounds were characterized using NMR, HRMS, IR and melting points. Evaluation of synthesized compounds against human colorectal (HCT116) and breast (CAL-51) cancer cell lines revealed potent antiproliferative activities. Nine compounds displayed GI₅₀ values in the low micromolar to submicromolar range; for example (E)-1-phenyl-3-(3,4,5-trimethoxyphenyl)prop-2-en-1-one (SSE14108) showed GI₅₀ of 0.473 ± 0.043 µM against HCT116 cells. Further analysis of these compounds revealed that (E)-3-(4-chlorophenyl)-1-phenylprop-2-en-1-one (SSE14105) and (E)-3-(4-methoxyphenyl)-1-phenylprop-2-en-1-one (SSE14106) caused rapid (4 and 8-h post-treatment) accumulation of p53 in HCT116 cells similar to its induction by positive control, Nutlin-3. Such activities were absent in 3-(4-methoxyphenyl)propiophenone (SSE14106H2) demonstrating the importance of conjugated ketone for antiproliferative and p53 stabilizing activity of the chalcones. We further evaluated p53 levels in the presence of cycloheximide (CHX) and the results showed that the p53 stabilization was regulated at post-translational level through blockage of its degradation. These chalcones can, therefore, act as fragment leads for further structure optimization to obtain more potent p53 stabilizing agents with enhanced anti-proliferative activities.     

Population size and structure of the African Softshell Turtle,Trionyx triunguis,in Dalaman,southwestern Turkey

We assessed the size of the population of the African Softshell Turtle, Trionyx triunguis, in Dalaman (Mu?la, Turkey), which is considered to be the largest population of the species in the Mediterranean, by using the Jolly-Seber mark-recapture method. A total of 415 individuals were caught during the summer months of 2009 to 2011, of which 148 were recaptures. From 267 marked individuals, 148 (55%) were male, 69 (26%) were female and 50 (19%) were juveniles and subadults of indeterminate sex. The male:female ratio was calculated to be 2.14:1. By using Model A' of the Jolly-Seber mark-recapture method, which includes deaths but no immigration, the mean population size in Kükürt and Küçükdalyan (Karg?n) lakes, together with Tersakan and Ta?l?çay creeks, were estimated to be 396±36. Based on this estimation, the population density was calculated to be 14 turtles/ha. Approximately 67.4% of the estimated population was marked. The mean capture probability (p) and mean survival ratio (Φ) were 0.094±0.009 and 0.957±0.076, respectively. Our results showed that the species’ population size is larger than previously estimated based on visual counts. The status “Vulnerable” C2a of the IUCN Red Data Book categories seems therefore more appropriate for the Mediterranean subpopulation of Trionyx triunguis than “Endangered” C2a.     

<Emphasis Type="Italic">Saccharothrix ghardaiensis</Emphasis> sp. nov., an actinobacterium isolated from Saharan soil

The taxonomic position of a new Saccharothrix strain, designated MB46^T, isolated from a Saharan soil sample collected in Mzab region (Ghardaïa province, South Algeria) was established following a polyphasic approach. The novel microorganism has morphological and chemical characteristics typical of the members of the genus Saccharothrix and formed a phyletic line at the periphery of the Saccharothrix espanaensis subcluster in the 16S rRNA gene dendrograms. Results of the 16S rRNA gene sequence comparisons revealed that strain MB46^T shares high degrees of similarity with S. espanaensis DSM 44229^T (99.2%), Saccharothrix variisporea DSM 43911^T (98.7%) and Saccharothrix texasensis NRRL B-16134^T (98.6%). However, the new strain exhibited only 12.5–17.5% DNA relatedness to the neighbouring Saccharothrix spp. On the basis of phenotypic characteristics, 16S rRNA gene sequence comparisons and DNA-DNA hybridizations, strain MB46^T is concluded to represent a novel species of the genus Saccharothrix, for which the name Saccharothrix ghardaiensis sp. nov. (type strain MB46^T = DSM 46886^T = CECT 9046^T) is proposed.     

VIM‐1 carbapenemase‐producing Escherichia coli in gulls from southern France

Acquired carbapenemases currently pose one of the most worrying public health threats related to antimicrobial resistance. A NDM‐1‐producing Salmonella Corvallis was reported in 2013 in a wild raptor. Further research was needed to understand the role of wild birds in the transmission of bacteria resistant to carbapenems. Our aim was to investigate the presence of carbapenem‐resistant Escherichia coli in gulls from southern France. In 2012, we collected 158 cloacal swabs samples from two gull species: yellow‐legged gulls (Larus michahellis) that live in close contact with humans and slender‐billed gulls (Chroicocephalus genei) that feed at sea. We molecularly compared the carbapenem‐resistant bacteria we isolated through culture on selective media with the carbapenem‐susceptible strains sampled from both gull species and from stool samples of humans hospitalized in the study area. The genes coding for carbapenemases were tested by multiplex PCR. We isolated 22 carbapenem‐resistant E. coli strains from yellow‐legged gulls while none were isolated from slender‐billed gulls. All carbapenem‐resistant isolates were positive for bla_VIM‐1 gene. VIM‐1‐producing E. coli were closely related to carbapenem‐susceptible strains isolated from the two gull species but also to human strains. Our results are alarming enough to make it urgently necessary to determine the contamination source of the bacteria we identified. More generally, our work highlights the need to develop more bridges between studies focusing on wildlife and humans in order to improve our knowledge of resistant bacteria transmission routes.     

The relationship between subclinical atherosclerosis and electrocardiographic abnormalities as biomarkers of cardiovascular risk

Electrocardiographic findings indicating myocardial disease, such as left ventricular hypertrophy or ST-T wave abnormalities, or the presence of coronary artery calcium, indicating atherosclerotic coronary artery disease, are both biomarkers of future cardiovascular (CV) risk. Although the risk factors for myocardial and coronary artery disease are similar, their concomitant expression has implications for CV disease screening and prevention programmes. The relationship between the resting 12-lead ECG and subclinical atherosclerosis measured as coronary artery calcium (CAC) with electron beam tomography was examined in 937 healthy participants (aged 40-50 years) enrolled in a CV risk screening study. Electrocardiograms and CAC were interpreted in blinded fashion, using standard criteria. An abnormal ECG was coded in 268 (28.6%) participants, most commonly left ventricular hypertrophy (3.1%), delayed precordial R wave transition (5.7%), T-wave abnormalities (10.0%) and intraventricular conduction delay (10.4%). Although abnormal ECG findings were associated with CV risk variables, the prevalence of any CAC was similar in subjects with any ECG finding (43 of 268, 16.0%) compared with those with normal ECGs (125 of 669, 18.7%, p=NS). In a logistic model controlling for CV risk factors including systolic blood pressure, low-density lipoprotein cholesterol (LDL-C), body mass index (BMI), glycosylated haemoglobin, race, age and gender, significant associations with CAC were found for LDL-C, race and BMI. There was no significant relationship between CAC and ECG abnormalities (odds ratio 0.80, 95% confidence interval 0.54-1.20). In conclusion, electrocardiographic abnormalities and subclinical calcified atherosclerosis were not significantly associated with each other in this middle-aged screening population. This suggests these two biomarkers may be complementary towards broader detection of latent CV risk.     

Negative selection of chronic lymphocytic leukaemia cells using a bifunctional rosette-based antibody cocktail

Background  

High purity of tumour samples is a necessity for accurate genetic and expression analysis and is usually achieved by positive selection in chronic lymphocytic leukaemia (CLL).