A UKST survey of blue objects towards the galactic centre - a search for early-type stars

We have begun a search for early-type stars towards the galactic centre which are potentially young objects situated within the inner few kiloparsecs of the disk. U and V (or I) band photographic photometry from the UK Schmidt Telescope has been obtained to identify the bluest candidates in nineteen Schmidt fields (centred close to the galactic centre). We have spectroscopically observed these targets for three fields with the FLAIR multi-fibre system to determine their spectral types. In particular; ten early B-type stars have been identified and equivalent width measurements of their Balmer and HeI lines have been used to estimate atmospheric parameters. These early-type objects have magnitudes in the range 11.5 less than or equal to V less than or equal to 16.0, and our best estimates of their distance (given probable highly variable reddening in this direction together with errors in the plate photometry) suggest that some of them originated close to (i.e R-g

New identifications for blue objects towards the Galactic center: post-AGE stars, Be/disk stars and others

As part of a programme to investigate spatial variations in the Galactic chemical composition, we have been searching for normal B-type stars and A-type supergiants near the Galactic center. During this search we have found eleven peculiar stars, and in some cases performed detailed abundance analyses of them which suggest that they may be at a post-AGB evolutionary stage.

LS 4825: A blue supergiant on the far side of the galaxy

We present high-resolution spectroscopic observations of LS 4825, a V = 12 B-type star in the Galactic center direction. On the basis of its stellar and interstellar spectra, we infer that it is likely to be a young supergiant at a distance of 21 +/- 5 kpc, and hence lying on the far side of the 'Galaxy. Adopting this hypothesis, a differential abundance analysis shows LS 4825 to have a chemical composition that is consistent with local B-type supergiants. These observations therefore represent the first detailed investigation of a star on the far side of the Galactic center. We trace multiple interstellar components in Ca II K and Na I D spectra, with velocities -206 less than or equal to v(lst) less than or equal to +93 km s(-1). We consider the likely origin of this gas and find that some components appear to trace matter lying close to the Galactic center. We discuss the possible use of such sight lines in furthering our understanding both of the nature of gas around the Galactic center and of the abundance gradient of the Galaxy.

Metallicities of 4 blue supergiants near the Galactic centre

High resolution optical spectra of four blue supergiants (HD148422 B0.5 Ib; HD178487 B0.5 Ib; HD179407 B1 Ib, HD163522 B1 Ib) which lie within 4.5 kpc of the Galactic centre are presented. Careful differential LTE model atmosphere analyses are used to quantify the differences in photospheric metal abundances between these stars and MK spectral standards in the solar neighborhood. A detailed non-LTE model atmosphere analysis of one star (HD163522) confirms that the LTE differential abundances should be reliable, provided we use a comparison star with similar atmospheric parameters.

The incidences of trisomy 8, trisomy 9 and D20S108 deletion in polycythaemia vera: an analysis of blood granulocytes using interphase fluorescence in situ hybridization:an analysis of blood granulocytes using interphase fluorescence in situ hybridization

We have used interphase fluorescence in situ hybridization (IFISH) to detect trisomy 8, trisomy 9 and 20q deletion in circulating granulocytes from patients with polycythaemia vera (PV). Out of 64 PV patients, 15 (23%) exhibited an abnormality. Two patients had trisomy 9, three had trisomy 8 and 10 patients had hemizygous deletion of D20S108 (a locus in the 20q common deleted region). Aberrant nuclei ranged from 10% to 80% in these 15 cases. There was no correlation between the presence of a marker and sex, age, interval between presentation and IFISH analysis, neutrophil or platelet count or therapy. Conventional marrow cytogenetic karyotype results were available in 23 cases and there was concurrence between these and blood IFISH in 16 cases (13 normal and three with 20q/D20S108 deletion by both methods). Three patients with D20S108 deletion by IFISH were normal by previous marrow cytogenetic testing and four cases with 20q deletion by previous marrow cytogenetics had normal blood granulocytes according to IFISH. Thus, we confirm that trisomies 8 and 9 and deletion of 20q are diagnostically useful markers of PV. IFISH analysis of blood granulocytes is a practical method for detecting these markers, but as an adjunct to, not as a substitute for, conventional marrow cytogenetics.

Gross anatomy of the muscle systems of Fasciola hepatica as visualized by phalloidin-fluorescence and confocal microscopy

Neuropeptides, biogenic amines and acetylcholine are expressed abundantly within the nervous systems of parasitic flatworms, and are particularly evident in the innervation of the musculature. Such associations have implicated the nervous system in locomotion, host attachment and reproductive co-ordination. Information on the muscle systems of parasitic flatworms is generally sparse, in particular those muscles associated with the reproductive system, intestinal tract and attachment apparatus. Also, the use of sectioned material has left description of the 3-dimensional organization of the musculature largely unrecorded. Using fluorescein isothiocyanate (FITC)-labelled phalloidin as a site-specific probe for filamentous actin, applied to whole-mount preparations of adult Fasciola hepatica and examined by confocal scanning laser microscopy, the present work reports on the organization of the major muscle systems in this trematode parasite. A highly regular array of outer circular, intermediate longitudinal and inner diagonal fibres distinguishes the body wall musculature, which is also involved in the development of both ventral and oral suckers. Circular fibres dominate the duct walls of the male and female reproductive systems, whereas the muscles of the intestinal tract have a somewhat diffuse arrangement of fibres. An understanding of the structural complexity of the muscle systems of parasitic flatworms is considered as fundamental to the interpretation of results from physiological experiments.

RP-LC with fluorescence detection of amino acids in rat brain synaptosomes

For the first time, a simple and validated reversed-phase liquid chromatography (RP-LC) with fluorescence detection has been developed for the simultaneous analysis of glutamate (Glu), ?-aminobutyric acid (GABA), glycine (Gly) and taurine (Tau) in Wistar and tremor rats brain synaptosomes. The samples were separated on a C18 analytical column with gradient elution of methanol and 0.1 mol L-1 potassium acetate at a flow rate of 1 mL min-1. Total run time was approximately 25 min. All calibration curves exhibited good linearity (r 2 > 0.999) within test ranges. The reproducibility was estimated by intra-and inter-day assays and RSD values were less than 2.48%. The recoveries were between 96.32 and 105.21%. The method was successfully applied to the quantification of amino acids in Wistar and tremor rats brain synaptosomes. Through this developed protocol, the levels of Glu in hippocampal and prefrontal cortical synaptosomes of tremor rats were both significantly elevated than those of adult Wistar rats whereas significantly decreased concentrations of GABA and Gly were observed in the hippocampal region of tremor rats without evident difference in the prefrontal cortex between experimental and control groups. In addition, our studies also showed a marked elevation of Tau in tremor rats hippocampal synaptosomes although there was no pronounced difference in the prefrontal cortical region of Wistar and tremor rats.

A novel dual-fluorescence strategy for functionally validating microRNA targets in 3-prime untranslated regions: regulation of the inward rectifier potassium channel Kir2.1 by miR-212.

Gene targeting by microRNAs is important in health and disease. We developed a functional assay for identifying microRNA targets and applied it to the K+ channel Kir2.1 (KCNJ2) which is dysregulated in cardiac and vascular disorders. The 3'UTR was inserted downstream of the mCherry red fluorescent protein coding sequence in a mammalian expression plasmid. MicroRNA sequences were inserted into the pSM30 expression vector which provides enhanced green fluorescent protein as an indicator of microRNA expression. HEK293 cells were co-transfected with the mCherry-3'UTR plasmid and a pSM30-based plasmid with a microRNA insert. The principle of the assay is that functional targeting of the 3'UTR by the microRNA results in a decrease in the red/green fluorescence intensity ratio as determined by automated image analysis. The method was validated with miR-1, a known downregulator of Kir2.1 expression, and was used to investigate targeting of the Kir2.1 3'UTR by miR-212. Red/green ratio was lower in miR-212-expressing cells compared to non-targeting controls, an effect that was attenuated by mutating the predicted target site. MiR-212 also reduced inward rectifier current and Kir2.1 protein in HeLa cells. This novel assay has several advantages over traditional luciferase-based assays including larger sample size, amenability to time course studies and adaptability to high-throughput screening.

Fluorescence yields for 3lnl′ Rydberg series in ions belonging to the He isoelectronic sequence

Fluorescence yields are reported for 3lnl' Rydberg series members in He-like ions of N, O and Ne. Results are presented for singlet series members with n values between 3 and 9, i.e. up to the region of overlap with the states belonging to the 4l4l' manifold in these atoms. This data is required, for example, for the interpretation of charge-exchange experiments involving bare N, O and Ne nuclei. Fluorescence yields, averaged over all 3lnl' singlet states, larger than 50% are obtained from about n = 7.

Laterite as a disinfectant.

A groundwater programme monitoring flow and quality of a potable water spring in a slum district in Kampala, Uganda revealed that although latrines acted as the principal means of organic waste disposal for the 1000 plus people living in the spring’s catchment, levels of faecal indicator bacteria (TVC 45 Deg C) in spring discharge remained at or below detection during the dry season, despite the presence of high levels of chloride (45mg/l-56mg/l) and nitrate (23mg/l – 30mg/l NO3-N), indicating sewage impacts. A programme of column and batch testing of laterite underlying the area provided a means of investigating the soil’s attenuation capacity under more controlled conditions.
X-ray diffraction analyses revealed the laterite to be dominated by quartz and kaolinite with minor (<5% by volume) quantities of haematite. Batch studies revealed that over 99% of bacteriophage adsorbed to haematite in less than 5 minutes. By contrast batch tests on haematite-free soil samples from the Blue Hills in Australia showed that although they had comparable dominant mineralogy and iron coverage on their surfaces (determined from Energy dispersive X-ray fluorescence) they had negligible ability to adsorb H40/1.
Based on the results of the batch studies using natural soils, a programme of batch studies, undertaken using pure haematite showed the mineral to have an extremely high capacity to adsorb bacteriophage, and suggested that it was responsible for the levels of attenuation observed.
The results of column studies were in keeping with the findings of batch experiments. Injection of 20 pore volumes of 300 pfu/mL of the bacteriophage H40/1 into a 20mm diameter glass column packed with sand sized (Ø>500µm) laterite revealed that the column could irreversibly remove over 2.5 log10 bacteriophage over its 10cm length.
Importance:
Mineralogical and batch test data provide convincing evidence to show that laterite can potentially act as an inexpensive means of removing micro organisms from water. The material, particularly in nodular form, displays considerable potential to act as an alternative filter material to conventional quartz filter sands.

Development and validation of an ultrasensitive fluorescence planar waveguide biosensor for the detection of paralytic shellfish toxins in marine algae

Marine dinoflagellates of the genera Alexandrium are well known producers of the potent neurotoxic paralytic shellfish toxins that can enter the food web and ultimately present a serious risk to public health in addition to causing huge economic losses. Direct coastal monitoring of Alexandrium spp. can provide early warning of potential shellfish contamination and risks to consumers and so a rapid, sensitive, portable and easy-to-use assay has been developed for this purpose using an innovative planar waveguide device. The disposable planar waveguide is comprised of a transparent substrate onto which an array of toxin-protein conjugates is deposited, assembled in a cartridge allowing the introduction of sample, and detection reagents. The competitive assay format uses a high affinity antibody to paralytic shellfish toxins with a detection signal generated via a fluorescently labelled secondary antibody. The waveguide cartridge is analysed by a simple reader device and results are displayed on a laptop computer. Assay speed has been optimised to enable measurement within 15 min. A rapid, portable sample preparation technique was developed for Alexandrium spp. in seawater to ensure analysis was completed within a short period of time. The assay was validated and the LOD and CCß were determined as 12 pg/mL and 20 pg/mL respectively with an intra-assay CV of 11.3% at the CCß and an average recovery of 106%. The highly innovative assay was proven to accurately detect toxin presence in algae sampled from the US and European waters at an unprecedented cell density of 10 cells/L. © 2012 Elsevier B.V. All rights reserved.

Structural effects on the pH-dependent fluorescence of naphthalenic derivatives and consequences for sensing/switching

Naphthalenic compounds are a rich resource for designers of fluorescent sensing/switching/logic systems. The degree of internal charge transfer (ICT) character in the fluorophore excited states can vary from negligible to substantial. Naphthalene-1,8;4,5-diimides (11–13), 1,8-naphthalimides (16) and 4-chloro-1,8-naphthalimides (15) are of the former type. The latter type is represented by the 4-alkylamino-1,8-naphthalimides (1). Whether ICT-based or not, these serve as the fluorophore in ‘fluorophore-spacer-receptor’ switching systems where PET holds sway until the receptor is bound to H+. On the other hand, 4-dialkylamino-1,8-naphthalimides (3–4) show modest H+-induced fluorescence switching unless the 4-dialkylamino group is a part of a small ring (5). Electrostatic destabilization of a non-emissive twisted internal charge transfer (ICT) excited state is the origin of this behaviour. An evolution to the non-emissive twisted ICT excited state is responsible for the weak emission of the model compound 6 (and related structures 7 and 8) across the pH range. Twisted ICT excited states are also implicated in the switch 9 and its model compound 10, which are based on the 6-dialkylamino-3H-benzimidazo[2,1-a]benz[d,e]isoquinolin-3-one fluorophore.

Human aflatoxin albumin adducts quantitatively compared by ELISA, HPLC with fluorescence detection, and HPLC with isotope dilution mass spectrometry

Essential to the conduct of epidemiologic studies examining aflatoxin exposure and the risk of heptocellular carcinoma, impaired growth, and acute toxicity has been the development of quantitative biomarkers of exposure to aflatoxins, particularly aflatoxin B-1. In this study, identical serum sample sets were analyzed for aflatoxin-albumin adducts by ELISA, high-performance liquid chromatography (HPLC) with fluorescence detection (HPLC-f), and HPLC with isotope dilution mass spectrometry (IDMS). The human samples analyzed were from an acute aflatoxicosis outbreak in Kenya in 2004 (n = 102) and the measured values ranged from 0.018 to 67.0, nondetectable to 13.6, and 0.002 to 17.7 ng/mg albumin for the respective methods. The Deming regression slopes for the HPLC-f and ELISA concentrations as a function of the IDMS concentrations were 0.71 (r(2) = 0.95) and 3.3 (r(2) = 0.96), respectively. When the samples were classified as cases or controls, based on clinical diagnosis, all methods were predictive of outcome (P < 0.01). Further, to evaluate assay precision, duplicate samples were prepared at three levels by dilution of an exposed human sample and were analyzed on three separate days. Excluding one assay value by ELISA and one assay by HPLC-f, the overall relative SD were 8.7%, 10.5%, and 9.4% for IDMS, HPLC-f, and ELISA, respectively. IDMS was the most sensitive technique and HPLC-f was the least sensitive method. Overall, this study shows an excellent correlation between three independent methodologies conducted in different laboratories and supports the validation of these technologies for assessment of human exposure to this environmental toxin and carcinogen.