Generating Caenorhabditis elegans UNC-33 antigens to be used for the Synthesis of Polyclonal Antibodies

UNC-33 and its human homolog, CRMP2 (Collapsin Response Mediator Protein-2), have been demonstrated to be involved in neurodevelopment as well as neurodegenerative disorders, primarily Alzheimer’s Disease. However, the physiology and interactions of these associations are vague. In order to further understand UNC-33/CRMP2, our group decided to use molecular biology and work toward the production of polyclonal antibodies specific to C. elegans UNC-33. To do this, we utilized the GST tag Gene Fusion System and produced two antigens- UNC-33 amino acid 48 to 212 and UNC-33 amino acid 48 to131 (UNC-3348-212 and UNC-3348-131). During this process, parameters were developed for the efficient expression and purification of these polypeptides. Once an effective protocol was established, GST fused UNC-3348-212 and UNC-3348-131 were expressed, purified, and tested for purity multiple times. Overall, these procedures resulted in the production of 3.72 mg and 2.10 mg of GST fused to UNC-3348-212 and GST fused to UNC-3348-131, respectively. Currently, these purified polypeptides are being injected into laboratory animals for the generation of polyclonal antibodies for UNC-33 research

Essential Ocean Variables for Biogeochemical Observations

Ocean biogeochemical (BGC) processes are fundamental for several key ocean ecosystem functions and services. For example, carbon dioxide uptake and storage by chemical uptake and biological fixation in combination with transport to depth remove anthropogenic carbon from the atmosphere and counteract global warming. Another important example would be nutrient regeneration from organic matter remineralization that sustains ocean productivity and, hence, also seafood supply. Although many BGC processes are rooted in surface waters where primary productivity and ocean / atmosphere exchange takes place, the deep ocean contributes strongly to the cycling and sequestration of elements. Without a consideration of organic particle attenuation in the deep pelagic and rates of remineralization and accumulation at the vast deep seafloor, the efficiency of the biological carbon pump and the recycling of nutrients – and potential effects of global change – can hardly be quantified. Considering the relevance of the deep ocean, the Deep Ocean Observing Strategy ‘DOOS’ is revising the Essential Ocean Variables (EOVs) specified by the Global Ocean Observing System (GOOS) with a deep ocean perspective. The DOOS community recognized, that most GOOS BGC EOVs are also relevant for deep ocean observations. However, the bottom of the sea requires more attention to assess its contribution to ocean BGC. ‘Seafloor labile organic matter’ and ‘seafloor respiration’ are candidates for additional EOVs beyond the GOOS EOVs ‘oxygen’ and ‘particulate matter’. Online questionnaires are prepared to facilitate an expert revision of GOOS BGC EOVs in order to make specific suggestions for modifications to the responsible GOOS panel. The revision includes the target phenomena and spatiotemporal scales, the observing platforms and networks considered relevant, and the requirements of observations. This contribution introduces the revision approach and invites experts to take part

Working the when, where, and who of social context: The case of a traumatic injury narrative

This is a preprint of an article whose final and definitive form has been published in the Qualitative Research in Psychology © 2010 Taylor & Francis; Qualitative Research in Psychology is available online at: www.tandfonline.com ; DOI: 10.1080/14780880802571176Within qualitative research it is widely recognised that context matters. Despite this, in recent years a number of authors have observed a lack of contextual awareness in qualitative analysis. The purpose of this article is to analyse categorically and holistically the process of meaning making in relation to context using data generated during a series of interviews with an individual who encountered a chronic back injury. Drawing upon the work of Holstein and Gubrium (200412. Holstein , JA and Gubrium , JF . 2004 . “ Context: working it up, down and across ” . In Qualitative research practice , Edited by: Seale , C , Gobo , G , Gubrium , J and Silverman , D . 297 – 331 . London : Sage . View all references), we focus on questions of when and where to illustrate the locally unarticulated contextual alternatives that can come into play at specific times and places. In addition, we raise questions pertaining to who as a means of further understanding the significance of others in relation to the participant's experiences. In doing so, we illuminate the ways in which different contexts can shape the meaning of injury. After discussing the inherent problems associated with studying the notion of context, we close by suggesting that examining the ways that context might operate throughout the process of meaning making can be a useful analytical tool for qualitative researchers working within the domain of psychology

A Storable Mediatorless Electrochemical Biosensor for Herbicide Detection.

A novel mediatorless photo-bioelectrochemical sensor operated with a biofilm of the cyanobacterium Synechocystis PCC6803 wt. for herbicide detection with long term stability (>20 days) was successfully developed and tested. Photoanodic current generation was obtained in the absence of artificial mediators. The inhibitory effect on photocurrent of three commonly used herbicides (i.e., atrazine, diuron, and paraquat) was used as a means of measuring their concentrations in aqueous solution. The injection of atrazine and diuron into the algal medium caused an immediate photocurrent drop due to the inhibition of photosynthetic electron transport. The detected concentrations were suitable for environmental analysis, as revealed by a comparison with the freshwater quality benchmarks set by the Environmental Protection Agency of the United States (US EPA). In contrast, paraquat caused an initial increase (~2 h) of the photocurrent effect of about 200%, as this compound can act as a redox mediator between the cells and the anode. A relatively long-term stability of the biosensor was demonstrated, by keeping anodes colonized with cyanobacterial biofilm in the dark at 4 °C. After 22 days of storage, the performance in terms of the photocurrent was comparable with the freshly prepared biosensor. This result was confirmed by the measurement of chlorophyll content, which demonstrated preservation of the cyanobacterial biofilm. The capacity of this biosensor to recover after a cold season or other prolonged environmental stresses could be a key advantage in field applications, such as in water bodies and agriculture. This study is a step forward in the biotechnological development and implementation of storable mediatorless electrochemical biosensors for herbicide detection

Impact of an equality constraint on the class-specific residual variances in regression mixtures:a Monte Carlo simulation study

Regression mixture models are a novel approach to modeling the heterogeneous effects of predictors on an outcome. In the model-building process, often residual variances are disregarded and simplifying assumptions are made without thorough examination of the consequences. In this simulation study, we investigated the impact of an equality constraint on the residual variances across latent classes. We examined the consequences of constraining the residual variances on class enumeration (finding the true number of latent classes) and on the parameter estimates, under a number of different simulation conditions meant to reflect the types of heterogeneity likely to exist in applied analyses. The results showed that bias in class enumeration increased as the difference in residual variances between the classes increased. Also, an inappropriate equality constraint on the residual variances greatly impacted on the estimated class sizes and showed the potential to greatly affect the parameter estimates in each class. These results suggest that it is important to make assumptions about residual variances with care and to carefully report what assumptions are made

Pt(IV) Pro-Drugs with an Axial HDAC Inhibitor Demonstrate Multimodal Mechanisms Involving DNA Damage and a Poptosis Independent of Cisplatin Resistance in A2780/A2780cis Cells

Epigenetic agents such as histone deacetylase (HDAC) inhibitors are widely investigated for use in combined anticancer therapy and the co-administration of Pt drugs with HDAC inhibitors has shown promise for the treatment of resistant cancers. Coordination of an HDAC inhibitor to an axial position of a Pt(IV) derivative of cisplatin allows the combination of the epigenetic drug and the Pt chemotherapeutic into a single molecule. In this work we carry out mechanistic studies on the known Pt(IV) complex cis,cis,trans-[Pt(NH3)2Cl2(PBA)2] (B) with the HDAC inhibitor 4-phenylbutyrate (PBA) and its derivatives cis,cis,trans-[Pt(NH3)2Cl2(PBA)(OH)] (A), cis,cis,trans-[Pt(NH3)2Cl2(PBA)(Bz)] (C), and cis,cis,trans-[Pt(NH3)2Cl2(PBA)(Suc)] (D) (Bz = benzoate, Suc = succinate). The comparison of the cytotoxicity, effect on HDAC activity, reactive oxygen species (ROS) generation, γ-H2AX (histone 2A-family member X) foci generation and induction of apoptosis in cisplatin-sensitive and cisplatin-resistant ovarian cancer cells shows that A – C exhibit multimodal mechanisms involving DNA damage and apoptosis independent of cisplatin resistance

Studies of propene conversion over H-ZSM-5 demonstrate the importance of propene as an intermediate in methanol-to-hydrocarbons chemistry

Funding Information: Johnson Matthey plc. is thanked for supplying the ZSM-5 zeolite and for financial support through the provision of industrial CASE studentships in partnership with the EPSRC (APH (EP/P510506/1), AZ (EP/N509176/1)). Experiments at the ISIS Neutron and Muon Source were made possible by a beam time allocation from the Science and Technologies Facilities Council. 53 The resources and support provided by the UK Catalysis Hubviamembership of the UK Catalysis Hub consortium and funded by EPSRC grants EP/R026815/1 and EP/R026939/1 are gratefully acknowledged. This research has been performed with the use of facilities and equipment at the Research Complex at Harwell; the authors are grateful to the Research Complex for this access and support.Peer reviewedPublisher PD