Synthesis and biological evaluation of novel folic acid receptor-targeted, β-cyclodextrin-based drug complexes for cancer treatment

Drug targeting is an active area of research and nano-scaled drug delivery systems hold tremendous potential for the treatment of neoplasms. In this study, a novel cyclodextrin (CD)-based nanoparticle drug delivery system has been assembled and characterized for the therapy of folate receptor-positive [FR(+)] cancer. Water-soluble folic acid (FA)-conjugated CD carriers (FACDs) were successfully synthesized and their structures were confirmed by 1D/2D nuclear magnetic resonance (NMR), matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS), high performance liquid chromatography (HPLC), Fourier transform infrared spectroscopy (FTIR), and circular dichroism. Drug complexes of adamatane (Ada) and cytotoxic doxorubicin (Dox) with FACD were readily obtained by mixed solvent precipitation. The average size of FACD-Ada-Dox was 1.5-2.5 nm. The host-guest association constant Ka was 1,639 M-1 as determined by induced circular dichroism and the hydrophilicity of the FACDs was greatly enhanced compared to unmodified CD. Cellular uptake and FR binding competitive experiments demonstrated an efficient and preferentially targeted delivery of Dox into FR-positive tumor cells and a sustained drug release profile was seen in vitro. The delivery of Dox into FR(+) cancer cells via endocytosis was observed by confocal microscopy and drug uptake of the targeted nanoparticles was 8-fold greater than that of non-targeted drug complexes. Our docking results suggest that FA, FACD and FACD-Ada-Dox could bind human hedgehog interacting protein that contains a FR domain. Mouse cardiomyocytes as well as fibroblast treated with FACD-Ada-Dox had significantly lower levels of reactive oxygen species, with increased content of glutathione and glutathione peroxidase activity, indicating a reduced potential for Dox-induced cardiotoxicity. These results indicate that the targeted drug complex possesses high drug association and sustained drug release properties with good biocompatibility and physiological stability. The novel FA-conjugated β-CD based drug complex might be promising as an anti-tumor treatment for FR(+) cancer

Assessment of Class Mutation Operators for C++ with the MuCPP Mutation System

Context: Mutation testing has been mainly analyzed regarding traditional mutation operators involving structured programming constructs common in mainstream languages, but mutations at the class level have not been assessed to the same extent. This fact is noteworthy in the case of C++ despite being one of the most relevant languages including object-oriented features. Objective: This paper provides a complete evaluation of class operators for the C++ programming language. MuCPP, a new system devoted to the application of mutation testing to this language, was developed to this end. This mutation system implements class mutation operators in a robust way, dealing with the inherent complexity of the language. Method: MuCPP generates the mutants by traversing the abstract syntax tree of each translation unit with the Clang API, and stores mutants as branches in the Git version control system. The tool is able to detect duplicate mutants, avoid system headers, and drive the compilation process. Then, MuCPP is used to conduct experiments with several open-source C programs. Results: The improvement rules listed in this paper to reduce unproductive class mutants have a significant impact in the computational cost of the technique. We also calculate the quantity and distribution of mutants generated with class operators, which generate far fewer mutants than their traditional counterparts. Conclusions: We show that the tests accompanying these programs cannot detect faults related to particular object-oriented features of C++. In order to increase the mutation score, we create new test scenarios to kill the surviving class mutants for all the applications. The results confirm that, while traditional mutation operators are still needed, class operators can complement them and help testers further improve the test suite

Analysis of the Mechanisms of Heat Acclimatization : Comparison of Heat-tolerance between Japanese and Thai Subjects

In order to clarify the mechanisms of heat acclimatization to tropical climates by permanent residence, changes in oral temperature due to heat load were compared in 10 male subjects in Chiang Mai, Thailand (tropical region) and 10 male subjects in Nagasaki, Japan (temperate region). Mean annual ambient temperature is 16.6℃ in Nagasaki and 25.9℃ in Chiang Mai. The experiments for the Thai subjects were performed in Chiang Mai and those for the Japanese subjects in Nagasaki during each region\u27s hottest months. The constitutional characteristics of the Thai subjects were a little shorter and slightly leaner than the Japanese. After staying at rest in the experimental room at 32℃ and 35% of relative humidity for at least 30 min, the lower legs were immersed into a hot water bath of 43℃ for 30 min. Mean initial oral temperature was 37.06±0.07℃ in Japanese and 37.12±0.05℃ in Thai subjects (P>0.05). Oral temperature rose after heat load and reached to 37.54±0.06℃ and 37.59±0.06℃ in Japanese and Thai subjects (P>0.05), respectively. Although the inhabitants in Chiang Mai were expected to be more acclimatized to heat compared to those in Nagasaki, no significant difference in the oral temperature was found between two groups throughout the experiment. It is speculated that the same rise in oral temperature in both groups of subjects is attributed to a lower sweat rate and an increase of dry heat loss in Thai subjects. In future studies, not only core temperature but also skin temperatures (dry heat loss) and sweat rate (evaporative heat loss) should be measured and analyzed

How to design high quality acupuncture trials—a consensus informed by evidence

An international panel including patients, clinicians, researchers, acupuncture and surgery trialists, statisticians, and experts in clinical epidemiology and methodology have developed new guidance for randomised controlled trials in acupuncture. It addresses the most prevalent and critical concerns of current acupuncture trials and will help funding agencies, trial registers, and journal editors to evaluate the relevance, importance, and quality of submitted trial proposals and completed trial

Comparison of Internal Ribosome Entry Site (IRES) and Furin-2A (F2A) for Monoclonal Antibody Expression Level and Quality in CHO Cells

10.1371/journal.pone.0063247PLoS ONE85

Antimicrobial resistance among migrants in Europe: a systematic review and meta-analysis

BACKGROUND: Rates of antimicrobial resistance (AMR) are rising globally and there is concern that increased migration is contributing to the burden of antibiotic resistance in Europe. However, the effect of migration on the burden of AMR in Europe has not yet been comprehensively examined. Therefore, we did a systematic review and meta-analysis to identify and synthesise data for AMR carriage or infection in migrants to Europe to examine differences in patterns of AMR across migrant groups and in different settings. METHODS: For this systematic review and meta-analysis, we searched MEDLINE, Embase, PubMed, and Scopus with no language restrictions from Jan 1, 2000, to Jan 18, 2017, for primary data from observational studies reporting antibacterial resistance in common bacterial pathogens among migrants to 21 European Union-15 and European Economic Area countries. To be eligible for inclusion, studies had to report data on carriage or infection with laboratory-confirmed antibiotic-resistant organisms in migrant populations. We extracted data from eligible studies and assessed quality using piloted, standardised forms. We did not examine drug resistance in tuberculosis and excluded articles solely reporting on this parameter. We also excluded articles in which migrant status was determined by ethnicity, country of birth of participants' parents, or was not defined, and articles in which data were not disaggregated by migrant status. Outcomes were carriage of or infection with antibiotic-resistant organisms. We used random-effects models to calculate the pooled prevalence of each outcome. The study protocol is registered with PROSPERO, number CRD42016043681. FINDINGS: We identified 2274 articles, of which 23 observational studies reporting on antibiotic resistance in 2319 migrants were included. The pooled prevalence of any AMR carriage or AMR infection in migrants was 25·4% (95% CI 19·1-31·8; I2 =98%), including meticillin-resistant Staphylococcus aureus (7·8%, 4·8-10·7; I2 =92%) and antibiotic-resistant Gram-negative bacteria (27·2%, 17·6-36·8; I2 =94%). The pooled prevalence of any AMR carriage or infection was higher in refugees and asylum seekers (33·0%, 18·3-47·6; I2 =98%) than in other migrant groups (6·6%, 1·8-11·3; I2 =92%). The pooled prevalence of antibiotic-resistant organisms was slightly higher in high-migrant community settings (33·1%, 11·1-55·1; I2 =96%) than in migrants in hospitals (24·3%, 16·1-32·6; I2 =98%). We did not find evidence of high rates of transmission of AMR from migrant to host populations. INTERPRETATION: Migrants are exposed to conditions favouring the emergence of drug resistance during transit and in host countries in Europe. Increased antibiotic resistance among refugees and asylum seekers and in high-migrant community settings (such as refugee camps and detention facilities) highlights the need for improved living conditions, access to health care, and initiatives to facilitate detection of and appropriate high-quality treatment for antibiotic-resistant infections during transit and in host countries. Protocols for the prevention and control of infection and for antibiotic surveillance need to be integrated in all aspects of health care, which should be accessible for all migrant groups, and should target determinants of AMR before, during, and after migration. FUNDING: UK National Institute for Health Research Imperial Biomedical Research Centre, Imperial College Healthcare Charity, the Wellcome Trust, and UK National Institute for Health Research Health Protection Research Unit in Healthcare-associated Infections and Antimictobial Resistance at Imperial College London

Lipid phosphate phosphatase 3 enables efficient thymic egress

Lipid phosphate phosphatase 3 in endothelial and epithelial cells promotes efficient T cell emigration from the thymus to the periphery