OBSERVATIONS OF DEUTERATED CYANOACETYLENE IN DARK CLOUDS

We have observed DC3N and HC3N in a number of cold dust clouds in order to derive the degree of deuterium fractionation. We find that the ratio of DC3N to HC3N is large, at about 0.05 or more, and discuss the implications of this result for the synthesis of cyanoacetylene. The observations are most readily interpreted if the deuteration of HC3N is linked to that of cyclic C3H2, which is also observed to exhibit a large degree of deuterium fractionation. HC3N deuteration levels comparable with those we observed are found to he just compatible with the mechanism suggested by Howe & Millar, but with adjusted rate coefficients. Freeze-out on to grain surfaces is also considered, but produces widespread deuterium enhancement in many species. contrary to observed levels.

Using location-aware technology for learning Geography in a real digital space outside the classroom

The use of new mobile technologies is still in its infancy in many secondary schools and there is limited evidence of the educational and pedagogical benefits on pupils’ learning in the formal school context. This qualitative study focuses on the use of handheld devices to teach a topic in geography to an examination class. Action research combined with pupil observations and focus group interviews are used to capture the pupils’ experiences of using mediascapes. Activity Theory is used as a lens to structure the analysis of the data and to report on the cognitive and affective impact of m-learning on pupils’ academic performance in the topic. Increased attainment and the development of wider skills for lifelong learning were identified in the study. The adaptability of the majority of pupils to the technology resulted in increased levels of willingness to learn in this novel context.

Deep sequencing reveals predominant expression of miR-21 amongst the small non-coding RNAs in retinal microvascular endothelial cells

The retinal vascular endothelium is essential for angiogenesis and is involved in maintaining barrier selectivity and vascular tone. The aim of this study was to identify and quantify microRNAs and other small regulatory non-coding RNAs (ncRNAs) which may regulate these crucial functions. Primary bovine retinal microvascular endothelial cells (RMECs) provide a well-characterized in vitro system for studying angiogenesis. RNA extracted from RMECs was used to prepare a small RNA library for deep sequencing (Illumina Genome Analyzer). A total of 6.8 million reads were mapped to 250 known microRNAs in miRBase (release 16). In many cases, the most frequent isomiR differed from the sequence reported in miRBase. In addition, five novel microRNAs, 13 novel bovine orthologs of known human microRNAs and multiple new members of the miR-2284/2285 family were detected. Several similar to 30 nucleotide sno-miRNAs were identified, with the most highly expressed being derived from snoRNA U78. Highly expressed microRNAs previously associated with endothelial cells included miR-126 and miR-378, but the most highly expressed was miR-21, comprising more than one-third of all mapped reads. Inhibition of miR-21 with an LNA inhibitor significantly reduced proliferation, migration, and tube-forming capacity of RMECs. The independence from prior sequence knowledge provided by deep sequencing facilitates analysis of novel microRNAs and other small RNAs. This approach also enables quantitative evaluation of microRNA expression, which has highlighted the predominance of a small number of microRNAs in RMECs. Knockdown of miR-21 suggests a role for this microRNA in regulation of angiogenesis in the retinal microvasculature. J. Cell. Biochem. 113: 20982111, 2012. (C) 2012 Wiley Periodicals, Inc.

A Critical Review of Screening Methods for the Detection of Chloramphenicol, Thiamphenicol, and Florfenicol Residues in Foodstuffs

This article gives an extensive overview of the wide range of analytical procedures developed for the detection of amphenicol antibiotic residues (chloramphenicol, thiamphenicol, and florfenicol) in many different types of foodstuffs (milk, meat, eggs, honey, seafood). Screening methods such as microbial inhibition methods, antibody-based immunoassays using conventional and biosensor-based detection systems, and some methods based on alternative recognition systems are described. The relative advantages and disadvantages of these methods are discussed and compared. The current status and future trends and developments in the need for accurate and rapid detection of this group of antimicrobials are also discussed.

Detection of unwanted residues of ivermectin in bovine milk by dissociation-enhanced lanthanide fluoroimmunoassay

Avermectins are frequently used to control parasitic infestations in many animal species. Previous studies have shown the long-term persistence of unwanted residues of these drugs in animal tissues and fluids. An immunoassay screening test for the detection acid quantification of ivermectin residues in bovine milk has been developed. After an extensive extraction procedure, milk samples were applied to a competitive dissociation-enhanced lanthanide fluoroimmunoassay using a monoclonal antibody against an ivermectin-transferrin conjugate, The monoclonal antibody, raised in Balb C mice, showed cross-reactivity with eprinomectin (92%), abamectin (82%) and doramectin (16%). The limit of detection of the assay (mean + 3 SD), calculated from the analysis of 17 known negative samples, was calculated as 4.6 ng/mL. Intra- and inter-assay RSDs were determined as 11.6% and 15.8%, respectively, using a negative bovine milk sample fortified with 25 ng/mL ivermectin. Six Friesian milking cows were treated with ivermectin, three with a pour-on formulation of the drug and three with an injectable solution at the manufacturer's recommended dose rate. An initial mean peak in ivermectin residue concentration was detected at day 4 (mean level = 47.5 ng/mL) and day 5 post-treatment (mean level = 26.4 ng/mL) with the injectable form and pour-on treatment, respectively. A second peak in residue concentration was observed using the DELFIA(R) procedure 28 days post-treatment in both treatment groups (23.1 ng/mL injectable and 51.9 ng/mL pour-on). These second peaks were not confirmed by HPLC and must at this Lime be considered to be false-positive results. By day 35 after treatment the mean ivermectin residue concentration of both groups fell below the limit of detection of the assay. Copyright (C) 2000 John Wiley & Sons, Ltd.

Development of a dual label time-resolved fluoroimmunoassay for the detection of beta-agonists in cattle urine

beta-Agonists are among the most widely abused drugs in veterinary medicine for the illegal promotion of farm animal growth. An array of analytical procedures has been developed to detect the residues of these compounds in many biological materials. As the number of beta-agonist formulations increases, it has become increasingly difficult to devise screening techniques capable of detecting a broad spectrum of these residues in a single test. A dual immunoassay based on time-resolved fluorescence was developed that incorporated a monoclonal antibody raised to tertiary butyl amines and a polyclonal antibody to biphenolic beta-agonists. This assay was capable of detecting residues of a range of beta-agonists present in bovine urine without the need for sample extraction. The limits of detection of the assay ranged from 1 to 8.5 ng ml(-1) depending on the cross-reactivity of individual compounds with the antibodies employed in the procedure.

Detection of ivermectin residues in bovine liver using an enzyme immunoassay

Ivermectin, a member of the avermectin group, is frequently used to control parasites in many food producing animal species. A method for the detection and quantification of ivermectin residues in bovine liver has been developed. Liver samples (4 g) were extracted with acetonitrile and applied to a competitive enzyme immunoassay using a polyclonal antiserum raised in rabbits against an ivermectin-transferrin conjugate, The limit of detection of the assay (mean +/- 3s) calculated from the analysis of 24 known negative samples was 1.6 ng g(-1), Intra- and inter-assay RSDs were determined as 8.8 and 14.6%, respectively, using a negative bovine liver sample fortified with 100 ng g(-1) of ivermectin. Four Friesian steers were treated with a pour-on application of ivermectin at a dose rate of 0.5 mg kg(-1) body mass then withdrawn and killed at 7, 14, 21 and 28 d, Livers mere removed and ivermectin residue concentrations determined using the proposed immunoassay procedure, Seven days post-treatment the ivermectin liver concentration was determined as 52.7 ng g(-1), decreasing to 4.1 ng(-1) at 28 d, All immunoassay results were confirmed using high-performance liquid chromatography (HPLC), The immunoassay and HPLC results for invermectin ranged from 1 to 58 ng g(-1) and were in close correlation (r = 0.99).

PEPTIDES RELATED TO THE DIPLOPTERA-PUNCTATA ALLATOSTATINS IN NONARTHROPOD INVERTEBRATES - AN IMMUNOCYTOCHEMICAL SURVEY

The allatostatins are a family of peptides isolated originally from the cockroach, Diploptera punctata. Related peptides have been identified in Periplaneta americana and the blowfly, Calliphora vomitoria. These peptides have been shown to be potent inhibitors of juvenile hormone synthesis in these species. A peptide inhibitor of juvenile hormone biosynthesis has also been isolated from the moth, Manduca sexta; however, this peptide has no structural homology with the D. punctata-type allatostatins. Investigations of the phylogeny of the D. punctata allatostatin peptide family have been started by examining a number of nonarthropod invertebrates for the presence of allatostatin-like molecules using immunocytochemistry with antisera directed against the conserved C-terminal region of this family. Allatostatin-like immunoreactivity (ALIR) was demonstrated in the nervous systems of Hydra oligactis (Hydrozoa), Moniezia expansa (Cestoda), Schistosoma mansoni (Trematoda), Artioposthia triangulata (Turbellaria), Ascaris suum (Nematoda), Lumbricus terrestris (Oligochaeta), Limax pseudoflavus (Gastropoda), and Eledone cirrhosa (Cephalopoda). ALIR could not be demonstrated in Ciona intestinalis (Ascidiacea). These results suggest that molecules related to the allatostatins may play an important role in nervous system function in many invertebrates as well as in insects and that they also have an ancient evolutionary lineage. (C) 1994 Wiley-Liss, Inc.

Nuclear factor-kappaB as a potential therapeutic target for the novel cytotoxic agent LC-1 in acute myeloid leukaemia

Nuclear factor-kappaB (NF-kappaB) has been implicated in a number of malignancies and has been suggested to be a potential molecular target in the treatment of leukaemia. This study demonstrated the constitutive activation of NF-kappaB in human myeloid blasts and a clear correlation between NF-kappaB expression and in vitro cytoprotection. High NF-kappaB expression was found in many of the poor prognostic acute myeloid leukaemia (AML) subtypes, such as French-American-British classification M0 and M7, and the poor cytogenetic risk group. The in vitro effects of LC-1, a novel dimethylamino-parthenolide analogue, were assessed in 62 primary untreated AML samples. LC-1 was found to be cytotoxic to AML cells in a dose-dependent manner, mediated through the induction of apoptosis. The median drug concentration necessary to kill 50% of the cells was 4.5 micromol/l for AML cells, compared with 12.8 micromol/l for normal marrow cells. LC-1 was shown to reduce the five individual human NF-kappaB Rel proteins in a dose-dependent manner. The subsequent inhibition of many NF-kappaB-regulated cytokines was also demonstrated. Importantly, sensitivity to LC-1 was correlated with the basal NF-kappaB activity. Consequently, LC-1 treatment provides a proof of principle for the use of NF-kappaB inhibitors in the treatment of AML.

Survival and persistence of opportunistic Burkholderia species in host cells

Burkholderia are microorganisms that have a unique ability to adapt and survive in many different environments. They can also serve as biopesticides and be used for the biodegradation of organic compounds. Usually harmless while living in the soil, these bacteria are opportunistic pathogens of plants and immunocompromised patients, and occasionally infect healthy individuals. Some of the species in this genus can also be utilised as biological weapons. They all possess very large genomes and have two or more circular chromosomes. Their survival and persistence, not only in the environment but also in host cells, offers a remarkable example of bacterial adaptation.

Variable selection via RIVAL and its application in nuclear material detection

In many situations, the number of data points is fixed, and the asymptotic convergence results of popular model selection tools may not be useful. A new algorithm for model selection, RIVAL (removing irrelevant variables amidst Lasso iterations), is presented and shown to be particularly effective for a large but fixed number of data points. The algorithm is motivated by an application of nuclear material detection where all unknown parameters are to be non-negative. Thus, positive Lasso and its variants are analyzed. Then, RIVAL is proposed and is shown to have some desirable properties, namely the number of data points needed to have convergence is smaller than existing methods.

Timekeeping strategies operate independently from spatial and accuracy demands in beat-interception movements

The prevailing paradigm for researching sensorimotor synchronisation in humans involves finger tapping and temporal accuracy measures. However, many successful sensorimotor synchronisation actions require not only to be 'in time', but also to be in a predefined spatial position. Reaching this spatial position in many everyday actions often exceeds the average amplitude of a finger movement. The aim of this study is to address how people coordinate their movement to be in the right place at the right time when the scale of the movement varies. Does the scale of the movement and accuracy demands of the movement change the ability to accurately synchronise? To address these questions, a sensorimotor synchronisation task with three different inter-beat intervals, two different movement amplitudes and two different target widths was used. Our experiment demonstrated that people use different timing strategies-employing either a movement strategy (varying movement time) or a waiting strategy (keeping movement time constant) for large-scale movements. Those two strategies were found to be equally successful in terms of temporal accuracy and variability (spread of errors). With longer interval durations (2.5 and 3.5 s), variability of sensorimotor synchronisation performance increased (measured as the spread of errors). Analysing the data using the Vorberg and Wing (Handbook of perception and action. Academic Press, New York, pp 181-262, 1996) model shows a need to develop further existing timing models of sensorimotor synchronisation so that they could apply to large-scale movements, where different movement strategies naturally emerge.

Novel macrocyclic and acyclic cationic lipids for gene transfer: Synthesis and in vitro evaluation

The synthesis and in vitro evaluation of four cationic lipid gene delivery vectors, characterized by acyclic or macrocyclic, and saturated or unsaturated hydrophobic regions, is described. The synthesis employed standard protocols, including ring-closing metathesis for macrocyclic lipid construction. All lipoplexes studied, formulated from plasmid DNA and a liposome composed of a synthesized lipid, 1,2-dimyristoyl-sn-glycero-3-ethylphosphocholine (EPC), and either 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) or cholesterol as co-lipid, exhibited plasmid DNA binding and protection from DNase I degradation, and concentration dependent cytotoxicity using Chinese hamster ovary-K1 cells. The transfection efficiency of formulations with cholesterol outperformed those with DOPE, and in many cases the EPC/cholesterol control, and formulations with a macrocyclic lipid (+/- 10:1) outperformed their acyclic counterparts (+/- 3:1).

A curvilinear approach to the kinetic analysis of linoleate peroxidation in aqueous liposomes by 2,2'azobis(2-amidoinopropane) dihydrochloride

Lipid peroxidation is a common feature of many chemical and biological processes, and is governed by a complex kinetic scheme. A fundamental stage in kinetic investigations of lipid peroxidation is the accurate determination of the rate of peroxidation, which in many instances is heavily reliant on the method of finite differences. Such numerical approximations of the first derivative are commonly employed in commercially available software, despite suffering from considerable inaccuracy due to rounding and truncation errors. As a simple solution to this, we applied three empirical sigmoid functions (viz. the Prout-Tompkins, Richards & Gompertz functions) to data obtained from the AAPH-mediated peroxidation of aqueous linoleate liposomes in the presence of increasing concentrations of Trolox, evaluating the curve fitting parameters using the widely available Microsoft Excel Solver add-in. We have demonstrated that the five-parameter Richards' function provides an excellent model for this peroxidation, and when applied to the determination of fundamental rate constants, produces results in keeping with those available in the literature. Overall, we present a series of equations, derived from the Richards' function, which enables direct evaluation of the kinetic measures of peroxidation. This procedure has applicability not only to investigations of lipid peroxidation, but to any system exhibiting sigmoid kinetics.

Exploring Scale Effects of Best/Worst Rank Ordered Choice Data to Estimate Benefits of Tourism in Alpine Grazing Commons

In many environmental valuation applications standard sample sizes for choice modelling surveys are impractical to achieve. One can improve data quality using more in-depth surveys administered to fewer respondents. We report on a study using high quality rank-ordered data elicited with the best-worst approach. The resulting "exploded logit" choice model, estimated on 64 responses per person, was used to study the willingness to pay for external benefits by visitors for policies which maintain the cultural heritage of alpine grazing commons. We find evidence supporting this approach and reasonable estimates of mean WTP, which appear theoretically valid and policy informative. © The Author (2011).