Molecular modelling of human CYP1B1 substrate interactions and investigation of allelic variant effects on metabolism

Molecular modelling of human CYP1B1 based on homology with the mammalian P450, CYP2C5, of known three-dimensional structure is reported. The enzyme model has been used to investigate the likely mode of binding for selected CYP1B1 substrates, particularly with regard to the possible effects of allelic variants of CYP1B1 on metabolism. In general, it appears that the CYP1B1 model is consistent with known substrate selectivity for the enzyme, and the sites of metabolism can be rationalized in terms of specific contacts with key amino acid residues within the CYP1B1 heme locus. Further-more, a mode of binding interaction for the inhibitor, a-naphthoflavone, is presented which accords with currently available information. The current paper shows that a combination of molecular modelling and experimental determinations on the substrate metabolism for CYP1B1 allelic variants can aid in the understanding of structure-function relationships within P450 enzymes. (C) 2003 Elsevier Science Ireland Ltd. All rights reserved.

Evaluation of a pilot telemedicine network for accident and emergency work

A pilot accident and emergency. teleconsulting service was established in Scotland. It was based at the accident and emergency department of the main hospital in Aberdeen. There were three peripheral sites in rural Grampian (Peterhead, Turriff and Huntly) and one in the Shetland Isles. The videoconferencing equipment used was connected by ISDN at 384 kbit/s. During the 15 months of the study, 1998 videoconference calls were made, of which 402 (20%) calls were made to the accident and emergency department for clinical consultations. The majority of the clinical calls (95%) were made between 09:00 and 17:00, and more than 90% were completed within 20 min. During the majority of calls (87%) one or more X-ray images were transmitted. The majority of patients (89%) received treatment without transportation to the main centre in Aberdeen. The present study demonstrated that accident and emergency teleconsultations can be technically reliable, effective in reducing the number of patient transfers and acceptable to the referring clinicians. As a result, approximately pound1.5 million has been made available by the government to develop a national system for Scotland.

Valence-tautomeric RbMnFe Prussian blue analogues:composition and time stability investigation

Three different stoichiometric forms of RbMn[Fe(CN) ]y·zHO [x = 0.96, y = 0.98, z = 0.75 (1); x = 0.94, y = 0.88, z = 2.17 (2); x = 0.61, y = 0.86, z = 2.71 (3)] Prussian blue analogues were synthesized and investigated by magnetic, calorimetric, Raman spectroscopic, X-ray diffraction, and Fe Mössbauer spectroscopic methods. Compounds 1 and 2 show a hysteresis loop between the high-temperature (HT) Fe(S = 1/2)-CN-Mn(S = 5/2) and the low-temperature (LT) Fe(S = 0)-CN-Mn(S = 2) forms of 61 and 135 K width centered at 273 and 215 K, respectively, whereas the third compound remains in the HT phase down to 5 K. The splitting of the quadrupolar doublets in the Fe Mössbauer spectra reveal the electron-transfer-active centers. Refinement of the X-ray powder diffraction profiles shows that electron-transfer-active materials have the majority of the Rb ions on only one of the two possible interstitial sites, whereas nonelectron-transfer-active materials have the Rb ions equally distributed. Moreover, the stability of the compounds with time and following heat treatment is also discussed. © Wiley-VCH Verlag GmbH & Co. KGaA, 2009.

Investigation of the role of protein kinase C (PKC) in cytostasis induced by tumour promoting phorbol esters and related compounds

Protein kinase C (PKC) is considered to be the major receptor for tumour promoting phorbol esters such as 12-0- tetradecanoylphorbol-13-acetate (TPA). These agents evoke a plethora of biological effects on cells in culture. The growth of A549 human lung carcinoma cells maintained in medium fortified with 10% foetal calf serum (FCS) is arrested for 6 days by TPA and other biologically active phorbol esters. In the work described in this thesis, the hypothesis was tested that modulation of PKC activity is closely related to events pivotal for cytostasis to occur. The effect of several phorbol esters, of newly synthesized analogues of diacylglycerols (DAG) and of bryostatins (bryos) on cell growth and ability to modulate activity of PKC has been investigated.Determination of the subcellular distribution of PKC following treatment of cells with TPA and partial enzyme purification by non-denaturing poly-acrylamide gel electrophoresis revealed translocation of enzyme activity from cytosoUc to paniculate fraction. Chronic exposure of cells to TPA resulted in a time and concentration dependent degradation of enzyme activity. Synthetic DAG and DAG analogues, unable to arrest the growth of cells at non-toxic concentrations, were neither able to affect subcellular PKC distribution nor compete effectively for phorbol ester binding sites at physiologically relevant concentrations. Bryos 1,2,4 and 5, natural products, possessing antineoplastic activity in mice, elicited transient arrest of A549 cell growth in vitro. They successfully competed for phorbol ester receptors in A549 cells with exquisite affinity and induced a shift in sub-cellular PKC distribution, though not to the same extent as PTA. Enzyme down-regulation resulted from prolonged exposure of cells to nanomolar concentrations of bryos. In vivo studies demonstrated that neither PDBu nor bryo 1 was able to inhibit A549 xenograft growth in athymic mice. The growth of A549 cell populations cultured under conditions of serum-deprivation was inhibited only transiently by biologically active phorbol esters. Fortification of serum-free medium with EGF or fetuin was able to partially restore sensitivity to maintained growth arrest by PTA. PKC translocation to the paniculate cellular fraction and subsequent enzyme down-regulation, induced by TPA, occurred in a manner similar to that observed in serum-supplemented cells. However, total PKC activity and cytosolic phorbol ester binding potential were greatly reduced in the serum-deprived cell population. Western blot analysis using monospecific monoclonal antibodies revealed the presence of PKC-a in both A549 cell populations, with significantly reduced protein levels in serum- deprived cells. PKC-/9 was not detected in either cell population.

The investigation of vegetation change using remote sensing to detect and monitor migration of landfill gas

Decomposition of domestic wastes in an anaerobic environment results in the production of landfill gas. Public concern about landfill disposal and particularly the production of landfill gas has been heightened over the past decade. This has been due in large to the increased quantities of gas being generated as a result of modern disposal techniques, and also to their increasing effect on modern urban developments. In order to avert diasters, effective means of preventing gas migration are required. This, in turn requires accurate detection and monitoring of gas in the subsurface. Point sampling techniques have many drawbacks, and accurate measurement of gas is difficult. Some of the disadvantages of these techniques could be overcome by assessing the impact of gas on biological systems. This research explores the effects of landfill gas on plants, and hence on the spectral response of vegetation canopies. Examination of the landfill gas/vegetation relationship is covered, both by review of the literature and statistical analysis of field data. The work showed that, although vegetation health was related to landfill gas, it was not possible to define a simple correlation. In the landfill environment, contribution from other variables, such as soil characteristics, frequently confused the relationship. Two sites are investigated in detail, the sites contrasting in terms of the data available, site conditions, and the degree of damage to vegetation. Gas migration at the Panshanger site was dominantly upwards, affecting crops being grown on the landfill cap. The injury was expressed as an overall decline in plant health. Discriminant analysis was used to account for the variations in plant health, and hence the differences in spectral response of the crop canopy, using a combination of soil and gas variables. Damage to both woodland and crops at the Ware site was severe, and could be easily related to the presence of gas. Air photographs, aerial video, and airborne thematic mapper data were used to identify damage to vegetation, and relate this to soil type. The utility of different sensors for this type of application is assessed, and possible improvements that could lead to more widespread use are identified. The situations in which remote sensing data could be combined with ground survey are identified. In addition, a possible methodology for integrating the two approaches is suggested.

A structural investigation of the alkali metal site distribution within bioactive glass using neutron diffraction and multinuclear solid state NMR

The atomic-scale structure of Bioglass and the effect of substituting lithium for sodium within these glasses have been investigated using neutron diffraction and solid state magic angle spinning (MAS) NMR. Applying an effective isomorphic substitution difference function to the neutron diffraction data has enabled the Na-O and Li-O nearest-neighbour correlations to be isolated from the overlapping Ca-O, O-(P)-O and O-(Si)-O correlations. These results reveal that Na and Li behave in a similar manner within the glassy matrix and do not disrupt the short range order of the network former. Residual differences are attributed solely to the variation in ionic radius between the two species. Successful simplification of the 2 sites present will be intimately related to the release properties of the glass system in physiological fluids such as plasma and saliva, and hence to the bioactivity of the material. Detailed structural knowledge is therefore a prerequisite for optimizing material design.

A snapshot of the lives of women with polycystic ovary syndrome:a photovoice investigation

Polycystic ovary syndrome affects 6  percent of women. Symptoms include hirsutism, acne, and infertility. This research explores the impact of polycystic ovary syndrome on women's lives using photovoice. Nine participants photographed objects related to their quality of life and made diary entries explaining each photograph. Three themes emerged from thematic analysis of the diaries: control (of symptoms and polycystic ovary syndrome controlling their lives), perception (of self, others, and their situation), and support (from relationships, health care systems, and education). These findings illuminate positive aspects of living with polycystic ovary syndrome and the role pets and social networking sites play in providing support for women with polycystic ovary syndrome.

Residue-specific investigation of the relationship between oxidation and activity of a dual specificity phosphatase by mass spectrometry

Redox regulation of signalling pathways is critical in proliferation and apoptosis; redox imbalance can lead to pathologies such as inflammation and cancer. Vaccinia H1-related protein (VHR; DUSP3) is a dual-specificity phosphatase important in controlling MAP kinase activity during cell cycle. the active-site motif contains a cysteine that acts as a nucleophile during catalysis. We used VHR to investigate the effect of oxidation in vitro on phosphatase activity, with the aim of determining how the profile of site-specific modification related to catalytic activity. Recombinant human VHR was expressed in E. coli and purified using a GST-tag. Protein was subjected to oxidation with various concentrations of SIN-1 or tetranitromethane (TNM) as nitrating agents, or HOCl. the activity was assayed using either 3-O-methylfluorescein phosphate with fluorescence detection or PIP3 by phosphate release with malachite green. the sites of oxidation were mapped using HPLC coupled to tandem mass spectrometry on an ABSciex 5600TripleTOF following in-gel digestion. More than 25 different concentration-dependent oxidative modifications to the protein were detected, including oxidations of methionine, cysteine, histidine, lysine, proline and tyrosine, and the % oxidized peptide (versus unmodified peptide) was determined from the extracted ion chromatograms. Unsurprisingly, methionine residues were very susceptible to oxidation, but there was a significant different in the extent of their oxidation. Similarly, tyrosine residues varied greatly in their modifications: Y85 and Y138 were readily nitrated, whereas Y38, Y78 and Y101 showed little modification. Y138 must be phosphorylated for MAPK phosphatase activity, so this susceptibility impacts on signalling pathways. Di- and tri- oxidations of cysteine residues were observed, but did not correlate directly with loss of activity. Overall, the catalytic activity did not correlate with redox state of any individual residue, but the total oxidative load correlated with treatment concentration and activity. This study provides the first comprehensive analysis of oxidation modifications of VHR, and demonstrates both heterogenous oxidant effects and differential residue susceptibility in a signalling phosphatase.

An investigation into the techniques for detecting hedgehogs in a rural landscape

Various techniques and devices have been developed for the purpose of detecting wildlife but many only provide optimum results in particular habitats, for certain species or under ideal weather conditions. It is therefore advantageous to understand the efficiency and suitability of techniques under different scenarios. The effectiveness of methods for detecting rural Irish hedgehogs was investigated as part of a larger study in April 2008. Road kill sightings and questionnaires were employed to locate possible hedgehog sites. Six sites were subsequently selected, and in these areas trapping, spotlighting and foot print tunnels were employed to investigate whether hedgehogs were indeed in the surrounding landscape. Infrared thermal imagery was examined as a detection device. Trapping and infrared imagery failed to detect hedgehogs in areas where they had previously been recorded. Footprint tunnels proved to be unsuccessful in providing absolute proof of hedgehogs in an area. No single method of detection technique could be relied upon to conclude the presence of hedgehogs in an area. A combination of methods is therefore recommended. However, spotlighting was the most effective method, taking a mean of 4 nights to detect a hedgehog, in comparison to 48 nights if footprint tunnels were used as a sole method of detection. This was also suggested by rarefaction curves of these two detection techniques, where over a 48 night period hedgehogs were expected to be recorded 27 times through spotlighting and just 5 times in an equivalent period of footprint tunnel nights.

Gas hydrate investigation at the Amsterdam mud volcano

We investigated gas hydrate in situ inventories as well as the composition and principal transport mechanisms of fluids expelled at the Amsterdam mud volcano (AMV; 2,025 m water depth) in the Eastern Mediterranean Sea. Pressure coring (the only technique preventing hydrates from decomposition during recovery) was used for the quantification of light hydrocarbons in near-surface deposits. The cores (up to 2.5 m in length) were retrieved with an autoclave piston corer, and served for analyses of gas quantities and compositions, and pore-water chemistry. For comparison, gravity cores from sites at the summit and beyond the AMV were analyzed. A prevalence of thermogenic light hydrocarbons was inferred from average C1/C2+ ratios <35 and d13C-CH4 values of -50.6 per mil. Gas venting from the seafloor indicated methane oversaturation, and volumetric gas-sediment ratios of up to 17.0 in pressure cores taken from the center demonstrated hydrate presence at the time of sampling. Relative enrichments in ethane, propane, and iso-butane in gas released from pressure cores, and from an intact hydrate piece compared to venting gas suggest incipient crystallization of hydrate structure II (sII). Nonetheless, the co-existence of sI hydrate can not be excluded from our dataset. Hydrates fill up to 16.7% of pore volume within the sediment interval between the base of the sulfate zone and the maximum sampling depth at the summit. The concave-down shapes of pore-water concentration profiles recorded in the center indicate the influence of upward-directed advection of low-salinity fluids/fluidized mud. Furthermore, the SO42- and Ba2+ pore-water profiles in the central part of the AMV demonstrate that sulfate reduction driven by the anaerobic oxidation of methane is complete at depths between 30 cm and 70 cm below seafloor. Our results indicate that methane oversaturation, high hydrostatic pressure, and elevated pore-water activity caused by low salinity promote fixing of considerable proportions of light hydrocarbons in shallow hydrates even at the summit of the AMV, and possibly also of other MVs in the region. Depending on their crystallographic structure, however, hydrates will already decompose and release hydrocarbon masses if sediment temperatures exceed ca. 19.3°C and 21.0°C, respectively. Based on observations from other mud volcanoes, the common occurrence of such temperatures induced by heat flux from below into the immediate subsurface appears likely for the AMV.

Please Stop Rubbing Your Relationship In My Face(book): An Investigation of Online Romantic Social Comparison

It is well-documented that social networking sites such as Facebook set the stage for social comparison. Such comparison has been linked to a number of negative outcomes including envy, negative moods, and lower self-esteem. The present research aims to extend current understanding of online social comparison by investigating how it pertains to romantic relationships. I hypothesized that for individuals high in attachment anxiety (compared to those low in this construct), online romantic social comparison might be related to negative consequences—which, in the current project, was operationalized as lower mood/affect and state self-esteem. Further, I hypothesized that there would be an interaction between attachment anxiety and relationship insecurities on these negative outcomes, such that the expected difference of attachment anxiety would be more pronounced under conditions priming relationship insecurities, relative to a control condition. Two experiments were conducted, one of which focused on single individuals, and the second focusing on individuals who were themselves in dating relationships. The paradigms of each entailed experimental manipulation of a key relationship-related variable (for single individuals, pessimism for future relationships; for dating individuals, the presence or absence of rejection threat), subsequent exposure to romantic content from Facebook, and finally, measures of affect and state self-esteem. I discovered partial support for the hypothesis that some single individuals—particularly those with higher, rather than lower, attachment anxiety—do indeed report feeling more negative moods and lower state self-esteem following exposure to romantic online content, in contrast to single individuals who had instead viewed neutral online content. The association between attachment anxiety and negative outcome was especially pertinent if individuals had been primed to believe that their own future romantic prospects were grim, or if attention had been drawn to their singleness. Among dating individuals, less support for hypotheses was found; however, exploratory post-hoc analyses revealed a promising (albeit weak) trend indicating that reinvestigation of the current hypotheses would be prudent.

Investigations of the present factors affecting the population characteristics and relative abundance of nile perch (Lates niloticus) at selected trawling sites of western part of Lake Victoria, Uganda, Kagegi Gulf

This report is based on the investigation of the factors affecting population characteristics and relative abundance of Nile perch in selected sites of Kagegi Gulf Lake Victoria Uganda in the month of November 2006. Nine (9) stations were sampled at depth strata of 0-10, 10-20, 20-30 and 30-40 m the mean catch rates were as follows; 8.75±5.5, 4.77±2.3, 6.33±0.3 and 1.34±1.1 tonnes per square kilometer respectively. The catch rates differed at various depth levels with p-value of 0.2940 at 5% level of significance. Limnological parameters were temperature 25.15±0.28, 23.68±0.20, 24.74±0.13 and 25.3±0.20°C; pH of 8.0±0.00, 7.7±0.11; 7.66±0.33 and 6.32±0.14, dissolved oxygen 7.37±0.24, 6.44±0.30, 6.32±0.14 and 6.22±0.14 mg/l; Total nitrogen 589.82±97.2, 514.34±68.8, 690.44±257.8 and 809.03±45.02 µgL-respectively with a p-value of 0.4392 at 5% level of significance. Prey type of Nile perch indicated 65.2% of haplochromine in 0-10 m depth and other strata >10 metre were dominated by Caridina nilotica. Generally investigations indicated that the catch rates of Nile perch at Kagegi gulf in various depth strata probably depended on both the physical and chemical parameters mentioned above.

Mössbauer Spectroscopic Study andMagnetic Investigation of Iron(III) Complexes on a DendrimericBasis

The functionalization of the molecular surface of various dendrimer generations with a phosphorous core and external amine groups is obtained by converting those amine groups into the corresponding imines of salicylaldehyde creating multiple coordination sites for the iron atoms. Treatment with iron(III) chloride yields multinuclear iron(III) complexes on a dendrimeric basis. The obtained multinuclear molecular systems exhibit extremely high total spin values. The influence of the generation growth on this type of coordination compounds is investigated by Mossbauer spectroscopy and SQUIDmagnetometry.

Investigation on water and sediment quality variation from coast to off shore and its relation to bent hoses biodiversity: case study Bandar tahery

This paper deals with the qualification of water and sediment particularly those of benthos, as well as their interaction results, through careful laboratory researches within 24 experimental sites around the bank of Bandar Taheri, Persian Gulf water of Iranian borders, under 13th phase south Pars project. Samples were carried to the laboratories and careful experimental tests such as physical chemical, heavy metal, nitrate ammoniac, toc and other biologic tests including various type of benthos count were performed. Data gained through Shanon and Dankan statistical analyses were also studied to determine the water, sediment and pollution rate. Resulted information would classify the area as less polluted area which is rather away from critical environmental zone, another word the area could be liable to change to an undesired one while the density rates of principal metals follows the Fe>Mn>Zn>Cu>Pb>Cd>N pattern.

INVESTIGATION BY MASS SPECTROMETRY OF THE UBIQUITOME AND PROTEIN CARGO OF EXOSOMES DERIVED FROM MYELOID-DERIVED SUPPRESSOR CELLS

Exosomes released by myeloid-derived suppressor cells (MDSC) are 30 nm in diameter extracellular vesicles that have been shown to carry biologically active proteins as well as ubiquitin molecules. Ubiquitin is known to have many functions, including involvement in the formation of exosomes, although the exact role is highly contested. In the study reported here, the proteome and ubiquitome of MDSC exosomes has been investigated by bottom-up proteomics techniques. This report identifies more than 1000 proteins contained in the MDSC exosome cargo and 489 sites of ubiquitination in more than 300 ubiquitinated proteins based on recognition of glycinylglycine tagged peptides without antibody enrichment. This has allowed extensive chemical and biological characterization of the ubiquitinated cohort compared to that of the entire protein cargo to support hypotheses on the role of ubiquitin in exosomes.