Lipid-mediated membrane binding properties of Disabled-2

AbstractDisabled-2 (Dab2) is an adaptor protein involved in several biological processes ranging from endocytosis to platelet aggregation. During endocytosis, the Dab2 phosphotyrosine-binding (PTB) domain mediates protein binding to phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) at the inner leaflet of the plasma membrane. As a result of platelet activation, Dab2 is released from α-granules and associates with both the αIIbβ3 integrin receptor and sulfatide lipids on the platelet surface through its N-terminal region including the PTB domain (N-PTB), thus, modulating platelet aggregation. Thrombin, a strong platelet agonist, prevents Dab2 function by cleaving N-PTB within the two basic motifs required for sulfatide association, a reaction that is prevented when Dab2 is bound to these sphingolipids. We have characterized the membrane binding properties of Dab2 N-PTB using micelles enriched with Dab2 lipid ligands, sulfatides and PtdIns(4,5)P2. Remarkably, NMR spectroscopy studies suggested differences in lipid-binding mechanisms. In addition, we experimentally demonstrated that sulfatide- and PtdIns(4,5)P2-binding sites overlap in Dab2 N-PTB and that both lipids stabilize the protein against temperature-induced unfolding. We found that whereas sulfatides induced conformational changes and facilitated Dab2 N-PTB penetration into micelles, Dab2 N-PTB bound to PtdIns(4,5)P2 lacked these properties. These results further support our model that platelet membrane sulfatides, but not PtdIns(4,5)P2, protect Dab2 N-PTB from thrombin cleavage

Timing of Colonization of Caries-Producing Bacteria: An Approach Based on Studying Monozygotic Twin Pairs

Findings are presented from a prospective cohort study of timing of primary tooth emergence and timing of oral colonization of Streptococcus mutans (S. mutans) in Australian twins. The paper focuses on differences in colonization timing in genetically identical monozygotic (MZ) twins. Timing of tooth emergence was based on parental report. Colonization timing of S. mutans were established by plating samples of plaque and saliva on selective media at 3 monthly intervals and assessing colony morphology. In 25% of individuals colonization occurred prior to emergence of the first tooth. A significant proportion of MZ pairs (21%) was discordant for colonization occurring before or after first tooth emergence, suggesting a role of environmental or epigenetic factors in timing of tooth emergence, colonization by S. mutans, or both. These findings and further application of the MZ co-twin model should assist in development of strategies to prevent or delay infection with S. mutans in children

Evaluation of preparedness of healthcare student volunteers against Middle East respiratory syndrome coronavirus (MERS-CoV) in Makkah, Saudi Arabia : a cross-sectional study

Aim: To assess the knowledge and attitude of senior medical, dental, nursing and pharmacy students toward Middle East respiratory syndrome-corona virus (MERS-CoV) in Saudi Arabia. Subjects and methods: A cross-sectional survey using a 21-item questionnaire was conducted for a 3-month period from November 2015–January 2016 in Makkah, Saudi Arabia. The questionnaire was designed to evaluate students’ understanding and perception of MERS-CoV. An ANOVA test was used to determine the association of study discipline and academic year with the student knowledge score on MERS. Results: A total of 364 students were assessed during the study. The majority (62%) of the participants were in the 20–22-year age group. More than half (53%) were pharmacy students followed by (22%) medical students. More than two thirds (71%) of the participants were aware that MERS is caused by the coronavirus. More than half (59%) of the participants believed that MERS can be transmitted through direct or indirect contact with infected camels. A statistically significant association was reported between the study discipline and mean knowledge score (p < 0.0001) with medical students achieving an overall better knowledge score compared with students from other study disciplines. Conclusion: Overall, students had good knowledge about MERS epidemiology, transmission and the recommended protective measures. However, students expressed their reluctance to work in healthcare facilities with inadequate MERS infection control isolation policies

Policing gender mobilities: interrogating the ‘feminisation of migration’ to Europe

This article proposes a gendered critique of the European Neighbourhood Policy, a framework that, amongst other things, aims to facilitate the mobility of migrants to the EU from the bordering countries. We highlight the ambivalences of European gender and migration regimes, and we take issue with the celebration of the ‘feminisation of migration’. The former fails to offer opportunities to women to safely embark on autonomous migratory projects, the latter contributes to reproduce traditional gender biases in the countries of origin as well as of destination. We conclude by suggesting that the EU critique to emigration countries for failing to tackle women’s discrimination is less than persuasive when assessed vis-á-vis with the curtailment on women’s independent mobility across European borders

Satisfying positivity requirement in the Beyond Complex Langevin approach

The problem of finding a positive distribution, which corresponds to a given complex density, is studied. By the requirement that the moments of the positive distribution and of the complex density are equal, one can reduce the problem to solving the matching conditions. These conditions are a set of quadratic equations, thus Groebner basis method was used to find its solutions when it is restricted to a few lowest-order moments. For a Gaussian complex density, these approximate solutions are compared with the exact solution, that is known in this special case

Upravljanje otporno na kvarove modularnim prekidačko-reluktantnim strojem nadahnuto prirodom

Fault tolerance is an obligatory feature in safety critical applications (aeronautical, aerospace, medical and military applications, power plants, etc.), where loss of life, environmental disasters, equipment destructions or unplanned downtimes must be avoided. For such applications, a novel bio-inspired motion control system is proposed. All its three components (the switched reluctance machine, the power converter and the control system) are designed to be as fault tolerant as possible. This paper describes all these three fault tolerant components: the bio-inspired control system having self-healing capabilities, the power converter with an extra leg and the fault tolerant modular machine. The theoretical expectations and simulation results are validated by means of laboratory experiments.Otpornost na kvarove je nužnost u sigurnosno kritičnim aplikacijama (aeronautičke, zrakoplovne, medicinske i vojne aplikacije, elektrane itd.), gdje je potrebno izbjeći smrtne slučajeve, prirodne nepogode, uništenje opreme ili neplanirane prekide u radu. Za takve aplikacije, predložen je novi slijedni sustav nadahnut prirodom. Sve tri komponente (prekidačko-reluktantni stroj, pretvarač i sustav upravljanja) su projektirani da budu što je više moguće otporni na kvarove. Ovaj rad opisuje sve tri komponente: sustav upravljanja nadahnut prirodom sa samoliječećim svojstvima, pretvarač s dodatnom granom i modularni stroj otporan na kvarove. Teoretska očekivanja i simulacijski rezultati su provjereni laboratorijskim eksperimentima

Causes of hOCT1-dependent cholangiocarcinoma resistance to sorafenib and sensitization by tumor-selective gene therapy

Although the multi-tyrosine kinase inhibitor sorafenib is useful in the treatment of several cancers, cholangiocarcinoma (CCA) is refractory to this drug. Among other mechanisms of chemoresistance, impaired uptake through human organic cation transporter type 1 (hOCT1) (gene SLC22A1) has been suggested. Here we have investigated the events accounting for this phenotypic characteristic and have evaluated the interest of selective gene therapy strategies to overcome this limitation. Gene expression and DNA methylation of SLC22A1 were analyzed using intrahepatic (iCCA) and extrahepatic (eCCA) biopsies (Copenhagen and Salamanca cohorts; n = 132) and The Cancer Genome Atlas (TCGA)-CHOL (n = 36). Decreased hOCT1 mRNA correlated with hypermethylation status of the SLC22A1 promoter. Treatment of CCA cells with decitabine (demethylating agent) or butyrate (histone deacetylase inhibitor) restored hOCT1 expression and increased sorafenib uptake. MicroRNAs able to induce hOCT1 mRNA decay were analyzed in paired samples of TCGA-CHOL (n = 9) and Copenhagen (n = 57) cohorts. Consistent up-regulation in tumor tissue was found for miR-141 and miR-330. High proportion of aberrant hOCT1 mRNA splicing in CCA was also seen. Lentiviral-mediated transduction of eCCA (EGI-1 and TFK-1) and iCCA (HuCCT1) cells with hOCT1 enhanced sorafenib uptake and cytotoxic effects. In chemically induced CCA in rats, reduced rOct1 expression was accompanied by impaired sorafenib uptake. In xenograft models of eCCA cells implanted in mouse liver, poor response to sorafenib was observed. However, tumor growth was markedly reduced by cotreatment with sorafenib and adenoviral vectors encoding hOCT1 under the control of the BIRC5 promoter, a gene highly up-regulated in CCA. Conclusion: The reason for impaired hOCT1-mediated sorafenib uptake by CCA is multifactorial. Gene therapy capable of selectively inducing hOCT1 in tumor cells can be considered a potentially useful chemosensitization strategy to improve the response of CCA to sorafenib

Formulation and optimisation of novel transfersomes for sustained release of local anaesthetic

Objective: To investigate the effect of formulation parameters on the preparation of transfersomes as sustained‐release delivery systems for lidocaine and to develop and validate a new high‐performance liquid chromatography (HPLC) method for analysis. Method: Taguchi design of experiment (DOE) was used to optimise lidocaine‐loaded transfersomes in terms of phospholipid, edge activator (EA) and phospholipid : EA ratio. Transfersomes were characterised for size, polydispersity index (PDI), charge and entrapment efficiency (%EE). A HPLC method for lidocaine quantification was optimised and validated using a mobile phase of 30%v/v PBS (0.01 m) : 70%v/v Acetonitrile at a flow rate of 1 ml/min, detected at 255 nm with retention time of 2.84 min. The release of lidocaine from selected samples was assessed in vitro. Key findings: Transfersomes were 200 nm in size, with PDI ~ 0.3. HPLC method was valid for linearity (0.1–2 mg/ml, R2 0.9999), accuracy, intermediate precision and repeatability according to ICH guidelines. The %EE was between 44% and 56% and dependent on the formulation parameters. Taguchi DOE showed the effect of factors was in the rank order : lipid : EA ratio ˃ EA type ˃ lipid type. Optimised transfersomes sustained the release of lidocaine over 24 h. Conclusion: Sustained‐release, lidocaine‐loaded transfersomes were successfully formulated and optimised using a DOE approach, and a new HPLC method for lidocaine analysis was developed and validated