High-quality RNA extraction from copepods for Next Generation Sequencing: A comparative study

Despite the ecological importance of copepods, few Next Generation Sequencing studies (NGS) have been performed on small crustaceans, and a standard method for RNA extraction is lacking. In this study, we compared three commonly-used methods: TRIzol®, Aurum Total RNA Mini Kit and Qiagen RNeasy Micro Kit, in combination with preservation reagents TRIzol® or RNAlater®, to obtain high-quality and quantity of RNA from copepods for NGS. Total RNA was extracted from the copepods Calanus helgolandicus, Centropages typicus and Temora stylifera and its quantity and quality were evaluated using NanoDrop, agarose gel electrophoresis and Agilent Bioanalyzer. Our results demonstrate that preservation of copepods in RNAlater® and extraction with Qiagen RNeasy Micro Kit were the optimal isolation method for high-quality and quantity of RNA for NGS studies of C. helgolandicus. Intriguingly, C. helgolandicus 28S rRNA is formed by two subunits that separate after heat-denaturation and migrate along with 18S rRNA. This unique property of protostome RNA has never been reported in copepods. Overall, our comparative study on RNA extraction protocols will help increase gene expression studies on copepods using high-throughput applications, such as RNA-Seq and microarrays.

Extraction and radioimmunoassay quantitation of neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) from human dental pulp tissue

The measurement of neuropeptides in complex biological tissue samples requires efficient and appropriate extraction methods so that immunoreactivity is retained for subsequent radioimmunoassay detection. Since neuropeptides differ in their molecular mass, charge and hydrophobicity, no single method will suffice for the optimal extraction of various neuropeptides. In this study, dental pulp tissue was obtained from 30 human non-carious teeth. Of the three different neuropeptide extraction methods employed, boiling in acetic acid in the presence of protease inhibitors yielded the highest levels of neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP). High pressure liquid chromatography (HPLC) analysis of dental pulp tissue verified the authenticity of the neuropeptides extracted. © 2003 Elsevier Science Ltd. All rights reserved.

Determination of metformin in plasma using a new ion pair solid phase extraction technique and ion pair liquid chromatography

This article describes the development of the first ion pair solid phase extraction technique (IPSPE), which has been applied to the extraction of metformin from plasma samples. In addition an ion pair chromatographic method was developed for the specific HPLC determination of metformin. Several extraction and HPLC methods have been described previously for metformin, however, most of them did not solve the problems associated with the high polarity of this drug. Drug recovery in the developed method was found to be more than 98%. The limit of detection and limit of quantification was 3 and 5 ng/ml, respectively. The intraday and interday precision (measured by coefficient of variation, CV%) was always less than 9%. The accuracy (measured by relative error, R.E.%) was always less than 6.9%. Stability analysis showed that metformin is stable for at least 3 months when stored at -70degreesC. The method has been applied to 150 patient samples as part of a medication adherence study. (C) 2003 Elsevier B.V. All rights reserved.

Performance analysis of a feasible air-cycle refrigeration system for road transport

The performance of an air-cycle refrigeration unit for road transport, which had been previously reported, was analysed in detail and compared with the original design model and an equivalent Thermo King SL200 vapour-cycle refrigeration unit. Poor heat exchanger performance was found to be the major contributor to low coefficient of performance values. Using state-of-the-art, but achievable performance levels for turbomachinery and heat exchangers, the performance of an optimised air-cycle refrigeration unit for the same application was predicted. The power requirement of the optimised air-cycle unit was 7% greater than the equivalent vapour-cycle unit at full-load operation. However, at part-load operation the air-cycle unit was estimated to absorb 35% less power than the vapour-cycle unit. The analysis demonstrated that the air-cycle system could potentially match the overall fuel consumption of the vapour-cycle transport refrigeration unit, while delivering the benefit of a completely refrigerant free system.

Design, construction and testing of an air-cycle refrigeration system for road transport

The environmental attractions of air-cycle refrigeration are considerable. Following a thermodynamic design analysis, an air-cycle demonstrator plant was constructed within the restricted physical envelope of an existing Thermo King SL200 trailer refrigeration unit. This unique plant operated satisfactorily, delivering sustainable cooling for refrigerated trailers using a completely natural and safe working fluid. The full load capacity of the air-cycle unit at -20 °C was 7,8 kW, 8% greater than the equivalent vapour-cycle unit, but the fuel consumption of the air-cycle plant was excessively high. However, at part load operation the disparity in fuel consumption dropped from approximately 200% to around 80%. The components used in the air-cycle demonstrator were not optimised and considerable potential exists for efficiency improvements, possibly to the point where the air-cycle system could rival the efficiency of the standard vapour-cycle system at part-load operation, which represents the biggest proportion of operating time for most units.

Reality Shows in Chile, origin and road map. Historical-Communication Dialogues

Reality shows are TV programs which represent a format used in television nowadays; however, the observation practices of individual and/or group intimacy dates from thousands years ago. Sometimes this was driven by voyeurism or morbid fascination, some others, by the purpose of guarding, supervising and maintaining status quo. This work offers an alternative answer to the explanation of this type of TV program emergence and relates this appearance to a government procedure bound up with modern State terrorism which began at the end of the eighteenth century and has been recalled by different regimes until present days.

Gaining Access to Data Sources in Statutory Social Work Agencies: The Long and Winding Road

This paper details a researcher's experience of gaining access to three statutory social work agencies in order to conduct a study examining how social workers respond to family support cases and how parents and carers experience the intervention of social workers in these cases. The stages in gaining access are outlined, the gate-keepers involved at each stage are identified and some of the difficulties encountered are highlighted and discussed. The paper concludes that researchers need to give greater priority to access considerations and that social work agencies need to give greater priority to co-operation with researchers.