Fasciola hepatica:Histological demonstration of apoptosis in the reproductive organs of flukes of triclabendazole-sensitive and triclabendazole-resistant isolates, and in field-derived flukes from triclabendazole-treated hosts, using in situ hybridisation to visualise endonuclease-generated DNA strand breaks

Investigation of the triclabendazole (TCBZ) resistance status of populations of Fasciola hepatica in field cases of fasciolosis, where treatment failure has been reported, can be supported by histological examination of flukes collected from recently treated hosts. In TCBZ-sensitive flukes (TCBZ-S) exposed to TCBZ metabolites for 1-4. days in vivo, but not in TCBZ-resistant flukes (TCBZ-R), morphological changes suggestive of apoptosis occur in cells undergoing meiosis or mitosis in the testis, ovary and vitelline follicles. In order to verify or refute the contention that efficacy of TCBZ treatment is associated with apoptosis in the reproductive organs of flukes, histological sections of TCBZ-S (Cullompton isolate) flukes and TCBZ-R (Sligo isolate) flukes were subjected to the TdT-mediated dUDP nick end labelling (TUNEL) in situ hybridisation method, a commercially available test specifically designed to label endonuclease-induced DNA strand breaks associated with apoptosis. Additionally, sections of in vivo-treated and untreated flukes originating from field outbreaks of suspected TCBZ-S and TCBZ-R fasciolosis were labelled by the TUNEL method. It was found that in treated TCBZ-S flukes, strong positive labelling indicating apoptosis was associated with morphologically abnormal cells undergoing mitosis or meiosis in the testis, ovary and vitelline follicles. Background labelling in the positive testis sections was attributed to heterophagy of cell debris by the sustentacular tissue. The triggering of apoptosis was probably related to failure of spindle formation at cell division, supporting the contention that TCBZ inhibits microtubule formation. In treated TCBZ-R (Sligo Type 1) flukes, and in treated flukes from field outbreaks of suspected TCBZ-R fasciolosis, no significant labelling was observed, while sections of fluke derived from a field case of fasciolosis where TCBZ resistance was not suspected were heavily labelled. Light labelling was associated with the testis of untreated Cullompton (TCBZ-S) and Sligo Type 2 (TCBZ-R) flukes, which exhibit abnormal spermatogenesis and spermiogenesis, respectively. This was attributed to apoptosis and to heterophagy of effete germ line cells by the sustentacular tissue. It is concluded that demonstration of apoptosis by in situ hybridisation using the TUNEL method on sections of 1-4. days in vivo TCBZ-treated F. hepatica can contribute to the diagnosis of TCBZ resistance in field outbreaks of fasciolosis. © 2012 Elsevier B.V.

Assessing synchrony in groups:Are you measuring what you think you are measuring?

Behavioural synchrony has been a popular topic of research in group living animals, but has so far lacked a standard approach. Previous studies have varied greatly in the number of behavioural states they have considered and the size of groups investigated. Here, a model of behavioural synchrony was used to test four measures of synchrony commonly used (proportion observations 100% conforming, mean proportion of conforming individuals, Ruckstuhl's group mean and the kappa coefficient). The model used scan samples of the behaviour of laying hens, originally categorised in 10 different behavioural states, as a basis for determining the agents' probability of performing behaviour states. We systematically varied the group size and the number of behavioural states in the model. The measures calculated from the behaviour of the model agents were compared against a synchrony factor that determined the 'motivation' of agents in the model to conform to the behaviour of other agents, for model runs with different group sizes and behavioural categories. The results of the model suggest that, of the measures considered, the kappa coefficient is the most suitable measure of synchrony. The kappa coefficient was the only measure of the four tested to control for expected levels of synchrony. Expected levels of synchrony are sensitive to both the number of behaviour states being examined and the size of the group, therefore observed levels of synchrony should be compared against expected levels to provide meaningful standardised measures. © 2012 Elsevier B.V.

Controlled chemistry of Moisture Sensitive Reagents in Ionic Liquids

Many reactions involving phosphorus reagents require highly anhydrous and inert conditions for their successful implementation. In particular, the use of PCl3 and its derivatives for synthesis is often hampered by the inherent sensitivity of the materials themselves. Ionic liquids are emerging as green alternative solvents for a range of processes, and in particular have proven to be excellent media for highly sensitive phosphorus reagents without the need for anhydrous or inert conditions. Herein, we report the use of ionic liquids as both storage and reaction media which allows difficult and sensitive chemistry to be achieved in a more accessible manner.

Gold Nanoparticles-Coated SU-8 for Sensitive Fluorescence-Based Detections of DNA

SU-8 epoxy-based negative photoresist has been extensively employed as a structural material for fabrication of numerous biological microelectro-mechanical systems (Bio-MEMS) or lab-on-a-chip (LOC) devices. However, SU-8 has a high autofluorescence level that limits sensitivity of microdevices that use fluorescence as the predominant detection workhorse. Here, we show that deposition of a thin gold nanoparticles layer onto the SU-8 surface significantly reduces the autofluorescence of the coated SU-8 surface by as much as 81% compared to bare SU-8. Furthermore, DNA probes can easily be immobilized on the Au surface with high thermal stability. These improvements enabled sensitive DNA detection by simple DNA hybridization down to 1 nM (a two orders of magnitude improvement) or by solid-phase PCR with sub-picomolar sensitivity. The approach is simple and easy to perform, making it suitable for various Bio-MEMs and LOC devices that use SU-8 as a structural material.

Effects of Gold Nanoparticle and Enzyme Precipitation on Enhancement of Surface Plasmon Resonance Immunoassay: A Super Sensitive Measurement of Anti- Glutamic Acid Decarboxylase Antibody

Introduction: In this study, colloidal gold nanoparticle and precipitation of an insoluble product formed by HRP-biocatalyzed oxidation of 3,3'-diaminobenzidine (DAB) in the presence of H2O2 were used to enhance the signal obtained from the surface plasmon resonance biosensor.

Methods: The colloidal gold nanoparticle was synthesized as described by Turkevitch et al., and their surface was firstly functionalized with HS(CH2)11(OCH2CH2)3COOH (OEG3¬-COOH) by self assembling technique. Thereafter, those OEG3-COOH functionalized nanoparticles were covalently conjugated with horseradish peroxidase (HRP) and anti-IgG antibody (specific to the Fc portion of all human IgG subclasses) to form an enzyme-immunogold complex. Characterization was performed by several methods: UV-Vis absorption, dynamic light scattering (DLS), transmission electron microscopy (TEM) and FTIR. The as-prepared enzyme-immunogold complex has been applied in enhancement of SPR immunoassay. A sensor chip used in the experiment was constructed by using 1:10 molar ratio of HS(CH2)11(OCH2CH2)6COOH and HS(CH2)11(OCH2CH2)3OH. The capture protein, GAD65 (autoantigen) which is recognized by anti-GAD antibody (autoantibody) in the sera of insulin-dependent diabetes mellitus patients, was immobilized onto the 1:10 surface via biotin-streptavidin interaction.

Results and conclusions: In the research, we reported the influences of gold nanoparticle and enzyme precipitation on the enhancement of SPR signal. Gold nanoparticle showed its enhancement as being consistent with other previous studies, while the enzyme precipitation using DAB substrate was applied for the first time and greatly amplified the SPR detection. As the results, anti-GAD antibody could be detected at pg/ml level which is far higher than that of commercial ELISA detection kit. This study indicates another way to enhance SPR measurement, and it is generally applicable to other SPR-based immunoassays.

Optimising the locations of thermally sensitive equipment in an aircraft crown compartment

A Design of Experiments (DoE) analysis was undertaken to generate a list of configurations for CFD numerical simulation of an aircraft crown compartment. Fitted regression models were built to predict the convective heat transfer coefficients of thermally sensitive dissipating elements located inside this compartment. These are namely the SEPDC and the Route G. Currently they are positioned close to the fuselage and it is of interest to optimise the heat transfer for reliability and performance purposes. Their locations and the external fuselage surface temperature were selected as input variables for the DoE. The models fit the CFD data with values ranging from 0.878 to 0.978, and predict that the optimum locations in terms of heat transfer are when the elements are positioned as close to the crown floor as possible ( and ?min. limits), where they come in direct contact with the air flow from the cabin ventilation system, and when they are positioned close to the centreline ( and ?CL). The methodology employed allows aircraft thermal designers to optimise equipment placement in confined areas of an aircraft during the design phase. The determined models should be incorporated into global aircraft numerical models to improve accuracy and reduce model size and computational time. © 2012 Elsevier Masson SAS. All rights reserved.

pH-Sensitive brush polymer-drug conjugates by ring-opening metathesis copolymerization

Representing a new category of polymer-drug conjugates, brush polymer-drug conjugates were prepared by ring-opening metathesis copolymerization. Following judicious structural design, these conjugates exhibited well-shielded drug moieties, significant water solubility, well-defined nanostructures, and acid-triggered drug release.

Politically sensitive encounters: Ethnography, access and the benefits of ‘hanging out’

Negotiating politically sensitive research environments requires both a careful consideration of the methods involved, and a great deal of personal resolve. In drawing upon two distinct yet comparable fieldwork experiences this paper champions the benefits of ethnographic methods in seeking to gain positionality and research legitimacy amongst those identified as future research participants. The authors explore and discuss their use of the ethnographic concept of ‘hanging out’ in politically sensitive environments when seeking to negotiate access to potentially hard to reach participants living in challenging research environments. Through an illustrative examination of their experiences in researching commemorative rituals in Palestine and mental health in a Northern Irish prison, both authors reflect upon their use of hanging out when seeking to break down barriers and gain acceptance amongst their target research participants. Their involvement in a range of activities, not directly related to the overall aims of the research project, highlight a need for qualitative researchers to adopt a flexible research design, one that embraces serendipitous or chance encounters, when seeking to gain access to hard to reach research participants or when issues of researcher legitimacy are particularly pronounced such as is the case in politically sensitive research environments.

Pepsin in bronchoalveolar lavage fluid:a specific and sensitive method of diagnosing gastro-oesophageal reflux-related pulmonary aspiration

Gastro-oesophageal reflux (GOR)-related aspiration is associated with respiratory disease, but the current "gold standard" investigation, the lipid-laden macrophage index (LLMI), is flawed. A specific marker of GOR-related aspiration should originate in the stomach, but not the lung. An assay to detect gastric pepsin in the bronchoalveolar lavage (BAL) of children was developed and validated.