Investigation of biological matrices for novel biomarkers by modern mass spectrometric methods.

The primary objective was to introduce novel or develop existing techniques for the identification of new biomarkers within a range of biological matrices by modem mass spectrometric methods. Samples interrogated were hemodialysis concentrate, whole tissue sections and whole blood, with each having inherent challenges for use with mass spectrometry. Hence, published research has focused on other biological matrices or modes of detection for achieving the relevant aim. This current work overcame these issues by improving sample preparation including, the use of existing protocols for completely novel applications. Haemodialysate solution has proved most fruitful for identifying new candidate biomarkers. We have reproducibly detected 15 known and 6 novel uremic solutes within hemodialysate, a biological matrix previously deemed unsuitable for liquid chromatography/electrospray ionisation-mass spectrometry (LC/ESI-MS). This work included a validation of the novel methodology with stability and reproducibility investigations to test robustness. This highlighted a previously unrecorded thermally labile nature of some uremic solutes within the dialysate solution. A putative structural assignment has been made for 4 novel uremic solutes named, 5-(amino-1,2,-dihydroxy-ethyl)-3-nitrosooxy-[ 1,2,4]trioxine-3,6-diol, 2-(5,6-diamino-6-diazenyl-cyclohex-l-enyl)-2-hydroxy-acetimidic acid, N-[2-(7-hydroxy-3-methyl-ocatahydro-imidazo[ 1,5-alpha]pyridine-6-yl)-2-oxo-acetyl]-guanidine, and 3-(6-hydroxy-cyclohexa- 1,3-dienyl)-2-imino-3-oxopropionaldehyde. We have also identified that the chemical nature of solutes will dictate their removal during dialysis treatment and highly polar conventional biomarkers, urea and creatinine, are not representative of non-polar analyte excretion. This allows us to knowledgably suggest recommendations to improve future treatment modalities. The mass spectrometric analysis of whole tissue sections, in particular those that are paraffin embedded, pose a new range of challenges. Current MALDI matrices are unable to penetrate deep within tissue limiting their use to the tissue surface only. We have evaluated a range of novel dansylated MALDI matrices for this purpose that is detectable by fluorescence spectroscopy to aid in locating the matrix compound following application. Each dansylated MALDI matrix showed better penetration into the tissue sections, yet maintaining fluorescence detection, when compared to standard matrices CHCA, sinapinic acid and DHB. Of these novel matrices dansylhydrazine proved most successful in ionising proteins and peptides by forming a protonated molecule and related adducts. These additional mass shifted peaks, when included in a tryptic peptide database search, can improve the probability of the original protein/peptide identification. We now have the potential to obtain a total image of frozen tissue by using CHCA and dansylhydrazine in combination to ionise proteins/peptides at the surface or at depth, respectively. Further work is required for the preparation protocols with paraffin embedded sections for this total imaging principle to be applied. Finally we have illustrated the advantages of discovering novel haemoglobin variants in blood with a new ion mobility time-of-flight mass spectrometer, the Synapt HDMS system (Waters, MA, USA). We have identified a new variant that co-elutes with glycated haemoglobin peaks present in chromatograms used for conventional blood screening. Ion mobility technology and data extraction enhances the clarity of the results regarding multiple charging and variant characteristics. This enabled the exact determination of the amino acid substitution or mutation for the variant, with its assignment to a haemoglobin chain and the specific location within the chain

A good place to raise a family? Comparing parents, service-providers\u27, and media perspectives of the inner and outer suburban areas of Melbourne

In Australia, the suburbs have historically been the favoured place to raise children. However this is being challenged both by social change and government policy encouraging innerurban renewal. We examined how inner-urban areas compare with more traditional suburban locations as places to raise a family. Recognising that there are many influences on perceptions of place, we included the opinions of parents, service-providers and the media in the two locations.Research focused on two municipalities in Melbourne, one located >25km and the other <10km from the CBD. Themes were obtained and compared from in-depth interviews with parents, serviceproviders and analysis of municipality-specific and state-wide newspaper articles.Service provision was the only theme common at all levels of analysis. For all other themes, differences occurred between perspectives of service-providers, media and parents, as well as between the two residential locations. These in-depth snapshots on the challenges and rewards of raising children in different urban locations can help inform government in planning healthy neighbourhoods that better serve the needs of contemporary Australian families

Estimating the potential of beekeeping to alleviate household poverty in rural Uganda

<div><p>Robust evidence underpinning the role of beekeeping in poverty alleviation is currently lacking. This study estimated the production potential for beekeepers in Northern Uganda by quantifying current production assets (equipment and knowledge) and impact on rural income streams range of proposed interventions. Intervention scenarios evaluated the economic benefits to be derived from different hive types combined with year-round provision of a nectar source (<i>Calliandra calothyrsus</i>) planted at varying density. Findings show that the type and number of beehive combinations used influenced the amount of revenue streams generated by the beekeepers. Addition of 20 log hives increased incomes 10 times, 20 KTBs increased revenues 16 times and Langstroth 18 times. Adding <i>Calliandra</i> trees as a forage source to the baseline scenario yielded revenues up to 17.6 times higher than the baseline. Implying that good management plus the introduction of a reliable nectar source, to off-set dry season challenges (absconding), could improve beekeeping productivity in Northern Uganda. Further research is required to validate <i>in situ</i> the impact of modelled scenarios on both honey yield and other ecosystem service benefits.</p></div

Does quantitative heterogeneity of human fetal hemoglobin (Hb F) reveal friends or foes of KLF1 in globin gene switching?

The chemical heterogeneity of fetal hemoglobin (Hb F) due to variable ratios of the Gγ and Aγ globin subunits reflects genetic complexity because of common dimorphisms such as Hb F Sardegna (or Aγ75(E19) Ile>Thr; also known as AγT) in Caucasians, and common variants such the Gγ globin variant, Hb F Malta I (or Gγ117(G19) His>Arg) that is in tight linkage disequilibrium with the β globin variant Hb Valletta (or β87(F3) Thr>Pro) and is found in 1.8% of neonates from Malta.peer-reviewe

A promising Start? The Local Network Fund for Children and Young People: Interim Findings from the National Evaluation

This is a summary of the interim evaluation report of the National Evaluation of the Local Network Fund (LNF) for Children and Young People. It is based on data gathered during the first phase of the evaluation (between October 2002 to December 2003). A final report of the National Evaluation will be available early in 2005. A consortium of research organisations, led by the University of Hull and including BMRB Social Research, The University of York and the University of Sheffield were commissioned in August 2002 by the-then Children and Young People’s Unit (CYPU) to carry out the evaluation