Evaluation of individual protein errors in silver-stained two-dimensional gels

The relationship between spot volume and variation for all protein spots observed on large format 2D gels when utilising silver stain technology and a model system based on mammalian NSO cell extracts is reported. By running multiple gels we have shown that the reproducibility of data generated in this way is dependent on individual protein spot volumes, which in turn are directly correlated with the coefficient of variation. The coefficients of variation across all observed protein spots were highest for low abundant proteins which are the primary contributors to process error, and lowest for more abundant proteins. Using the relationship between spot volume and coefficient of variation we show it is necessary to calculate variation for individual protein spot volumes. The inherent limitations of silver staining therefore mean that errors in individual protein spot volumes must be considered when assessing significant changes in protein spot volume and not global error. (C) 2003 Elsevier Science (USA). All rights reserved.

Validation of assembled nucleic acid-based tests in diagnostic microbiology laboratories

Medical microbiology and virology laboratories use nucleic acid tests (NAT) to detect genomic material of infectious organisms in clinical samples. Laboratories choose to perform assembled (or in-house) NAT if commercial assays are not available or if assembled NAT are more economical or accurate. One reason commercial assays are more expensive is because extensive validation is necessary before the kit is marketed, as manufacturers must accept liability for the performance of their assays, assuming their instructions are followed. On the other hand, it is a particular laboratory's responsibility to validate an assembled NAT prior to using it for testing and reporting results on human samples. There are few published guidelines for the validation of assembled NAT. One procedure that laboratories can use to establish a validation process for an assay is detailed in this document. Before validating a method, laboratories must optimise it and then document the protocol. All instruments must be calibrated and maintained throughout the testing process. The validation process involves a series of steps including: (i) testing of dilution series of positive samples to determine the limits of detection of the assay and their linearity over concentrations to be measured in quantitative NAT; (ii) establishing the day-to-day variation of the assay's performance; (iii) evaluating the sensitivity and specificity of the assay as far as practicable, along with the extent of cross-reactivity with other genomic material; and (iv) assuring the quality of assembled assays using quality control procedures that monitor the performance of reagent batches before introducing new lots of reagent for testing.

Implications of seasonal climate forecasts on world wheat trade: a stochastic, dynamic analysis

Improvements in seasonal climate forecasts have potential economic implications for international agriculture. A stochastic, dynamic simulation model of the international wheat economy is developed to estimate the potential effects of seasonal climate forecasts for various countries' wheat production, exports and world trade. Previous studies have generally ignored the stochastic and dynamic aspects of the effects associated with the use of climate forecasts. This study shows the importance of these aspects. In particular with free trade, the use of seasonal forecasts results in increased producer surplus across all exporting countries. In fact, producers appear to capture a large share of the economic surplus created by using the forecasts. Further, the stochastic dimensions suggest that while the expected long-run benefits of seasonal forecasts are positive, considerable year-to-year variation in the distribution of benefits between producers and consumers should be expected. The possibility exists for an economic measure to increase or decrease over a 20-year horizon, depending on the particular sequence of years.

Correlating gene expression with larval competence and the effect of age and parentage on metamorphosis in the tropical abalone Haliotis asinina

The non-geniculate crustose coralline alga (CCA) Mastophora pacifica can induce the metamorphosis of competent Haliotis asinina (Vetigastropoda) larvae. The ability to respond to this natural cue varies considerably with larval age, with a higher proportion of older larvae (e.g. 90 h) able to metamorphose in response to M. pacifica than younger larvae (e.g. 66 h). Here we document the variation in time to acquisition of competence within a larval age class. For example, after 18 h of exposure to M. pacifica, approximately 15 and 36% of 84 and 90-h-old H. asinina larvae had initiated metamorphosis, respectively. This age-dependent response to M. pacifica is also observed when different aged larvae are exposed to CCA for varying periods. A higher proportion of older larvae require shorter periods of exposure to CCA than younger larvae in order to initiate metamorphosis. In this experiment, as in the previous, a small proportion of young larvae were able to respond to brief periods of CCA exposure, suggesting that they had developed the same state of competency as the majority of their older counterparts. Comparisons of the proportions of larvae undergoing metamorphosis between families reveals that parentage also has a significant (P < 0.05) affect on whether an individual will initiate metamorphosis at a given age. These familial differences are more pronounced when younger, largely pre-competent larvae (i.e. 66 h old) are exposed to M. pacifica, with proportions of larvae undergoing metamorphosis differing by as much as 10 fold between families. As these data suggest that variation in the rate of development of the competent state has a genetic basis, and as a first step towards identifying the molecular basis to this variation, we have identified numerous genes that are differentially expressed later in larval development using a differential display approach. Spatial expression analysis of these genes suggests that they may be directly involved in the acquisition of competence, or may play a functional role in the postlarva following metamorphosis.

Genomic organization of human arylamine N-acetyltransferase Type I reveals alternative promoters that generate different 5'-UTR splice variants with altered translational activities

In humans, a polymorphic gene encodes the drug-metabolizing enzyme NATI (arylamine N-acetyltransferase Type 1), which is widely expressed throughout the body. While the protein-coding region of NATI is contained within a single exon, examination of the human EST (expressed sequence tag) database at the NCBI revealed the presence of nine separate exons, eight of which were located in the 5'non-coding region of NATI. Differential splicing produced at least eight unique mRNA isoforms that could be grouped according to the location of the first exon, which suggested that NATI expression occurs from three alternative promoters. Using RT (reverse transcriptase)-PCR, we identified one major transcript in various epithelial cells derived from different tissues. In contrast, multiple transcripts were observed in blood-derived cell lines (CEM, THP-1 and Jurkat), with a novel variant, not identified in the EST database, found in CEM cells only. The major splice variant increased gene expression 9-11-fold in a luciferase reporter assay, while the other isoforrns were similar or slightly greater than the control. We examined the upstream region of the most active splice variant in a promoter-reporter assay, and isolated a 257 bp sequence that produced maximal promoter activity. This sequence lacked a TATA box, but contained a consensus Sp1 site and a CAAT box, as well as several other putative transcription-factor-binding sites. Cell-specific expression of the different NATI transcripts may contribute to the variation in NATI activity in vivo.

Pollen transport and deposition in riverine and marine environments within the humid tropics of northeastern Australia

Mechanisms of pollen transport in the humid tropics region of northeastern Australia were investigated to support the interpretation of a long Quaternary pollen record from ODP Site 820 located on the adjacent continental slope. Pollen analysis of surface sediment samples from the channels of two major river catchments demonstrated internal consistency in pollen spectra and little fluvial pollen sorting in relation to sediment variation. Differences in modem pollen spectra between catchments reflect existing variation in vegetation cover that, in turn, reflects climatic differences between catchments. Recent pollen spectra from top samples of the ODP core have sufficient in common with the riverine samples to suggest that the rivers are contributing a major pollen component to the offshore sediments, but these have been size sorted by marine action. Recent pollen samples from core tops taken from the Grafton Passage on the continental shelf that was thought to be the major passage for pollen transport to ODP Site 820 show significant differences to both riverine and ODP samples and suggest that pollen are dispersed across the continental shelf and through the outer Great Barrier Reef system in a more complex way than anticipated. (c) 2004 Elsevier B.V. All rights reserved.

Butyrylcholinesterase: Association with the metabolic syndrome and identification of 2 gene loci affecting activity

Background: Plasma cholinesterase activity is known to be correlated with plasma triglycerides, HDL- and LDL-cholesterol, and other features of the metabolic syndrome. A role in triglyceride metabolism has been proposed. Genetic variants that decrease activity have been studied extensively, but the factors contributing to overall variation in the population are poorly understood. We studied plasma cholinesterase activity in a sample of 2200 adult twins to assess covariation with cardiovascular risk factors and components of the metabolic syndrome, to determine the degree of genetic effects on enzyme activity, and to search for quantitative trait loci affecting activity. Methods and Results: Cholinesterase activity was lower in women than in men before the age of 50, but increased to activity values similar to those in males after that age. There were highly significant correlations with variables associated with the metabolic syndrome: plasma triglyceride, HDL- and LDL-cholesterol, apolipoprotein B and E, urate, and insulin concentrations; gamma-glutamyltransferase and aspartate and alanine aminotransferase activities; body mass index; and blood pressure. The heritability of plasma cholinesterase activity was 65%. Linkage analysis with data from the dizygotic twin pairs showed suggestive linkage on chromosome 3 at the location of the cholinesterase WHO gene and also on chromosome 5. Conclusions: Our results confirm and extend the connection between cholinesterase, cardiovascular risk factors, and metabolic syndrome. They establish a substantial heritability for plasma cholinesterase activity that might be attributable to variation near the structural gene and at an independent locus. (c) 2006 American Association for Clinical Chemistry.

Longitudinal Genetic Analysis of Plasma Lipids

The consensus from published studies is that plasma lipids are each influenced by genetic factors, and that this contributes to genetic variation in risk of cardiovascular disease. Heritability estimates for lipids and lipoproteins are in the range .48 to .87, when measured once per study participant. However, this ignores the confounding effects of biological variation measurement error and ageing, and a truer assessment of genetic effects on cardiovascular risk may be obtained from analysis of longitudinal twin or family data. We have analyzed information on plasma high-density lipoprotein (HDL) and low-density lipoprotein (LDL) cholesterol, and triglycerides, from 415 adult twins who provided blood on two to five occasions over 10 to 17 years. Multivariate modeling of genetic and environmental contributions to variation within and across occasions was used to assess the extent to which genetic and environmental factors have long-term effects on plasma lipids. Results indicated that more than one genetic factor influenced HDL and LDL components of cholesterol, and triglycerides over time in all studies. Nonshared environmental factors did not have significant long-term effects except for HDL. We conclude that when heritability of lipid risk factors is estimated on only one occasion, the existence of biological variation and measurement errors leads to underestimation of the importance of genetic factors as a cause of variation in long-term risk within the population. In addition our data suggest that different genes may affect the risk profile at different ages.

SUPORTE SOCIAL E AUTOEFICÁCIA EM IDOSOS

As questões do desenvolvimento humano desde tempos remotos são de interesse das diversas áreas do conhecimento. Tendo em vista o aumento da expectativa de vida, as questões relacionadas ao processo de envelhecimento ganharam importância, principalmente no que tange às políticas de saúde. A saúde é preocupação central no campo do envelhecimento, uma vez que problemas de saúde podem acarretar diversas dificuldades para os idosos e para suas famílias. Este estudo teve por objetivo buscar identificar as associações entre percepções de suporte social e autoeficácia em idosos. Suporte social e autoeficácia são variáveis estudadas em contextos diversos. Estudos revelam que ambas são capazes de colaborar para proteger e promover saúde. Suporte social diz respeito ao grupo de informações que leva o indivíduo a acreditar que é valorizado, amado e estimado e que o faz sentir-se pertencente a uma rede social de comunicação recíproca. Autoeficácia pode ser definida como o conjunto de crenças que o indivíduo tem sobre suas capacidades para organizar e executar ações para o cumprimento de tarefas específicas. Portanto, identificar a existência de associações entre as percepções de suporte social e autoeficácia pode contribuir para dar maior solidez aos conhecimentos já existentes sobre esses temas e sobre o processo de envelhecimento. Participaram do estudo 61 idosos de ambos os sexos, com idade média de 67 anos, com grau de escolaridade que variou entre ensino fundamental até pós-graduação e participantes em ações de promoção à saúde. Todos foram voluntários e autorizaram sua participação por meio da assinatura de termo de consentimento. A coleta de dados foi feita mediante a aplicação de questionário contendo escalas para avaliar os construtos, além de questões para caracterização dos participantes. Os dados foram analisados eletronicamente por meio do cálculo de frequências, percentuais, médias e desvios padrão. Para se testar a hipótese, foi calculada a correlação (r de Pearson) entre as variáveis do estudo. Entre os resultados obtidos, a percepção de suporte social, tanto na dimensão emocional quanto na dimensão prática, para os idosos participantes deste estudo, é de que muitas vezes são amados, cuidados, estimados, valorizados e, que pertencem a uma rede de comunicação e obrigações mútuas. Em se tratando da percepção da autoeficácia, constatou-se que esses participantes possuem crenças de que às vezes é verdade que são eficazes e que conseguem organizar e executar ações para o cumprimento de tarefas específicas. Aponta-se ainda que não existem correlações significativas entre percepção de suporte social e autoeficácia, ou seja, os níveis de percepção de suporte social não variam proporcionalmente à variação das crenças de autoeficácia em idosos. Isto significa dizer que para estes participantes, não seria eficaz investir em melhorias do suporte social, pois isto não teria impacto em suas crenças de autoeficácia. Indica-se a realização de novas pesquisas com idosos não participantes de ações de promoção à saúde, pois se pode pensar que idosos não engajados nestes tipos de ações poderiam necessitar de maior suporte social e assim, terem suas crenças de autoeficácia associadas à sua percepção de suporte social.

A study of inputs and their influence on technological innovative activity

Technological innovation has been widely studied: however surprisingly little is known about the experience of managing the process. Most reports tend to be generalistic and/or prescriptive whereas it is argued that multiple sources of variation in the process limit the value of these. A description of the innovation process is given together with a presentation of what is knovrn from existing studies. Gaps identified in this area suggest that a variety of organisational influences are important and an attempt is made to identify some of these at individual, group and organisational level. A simple system model of the innovation management process is developed. Further investigation of the influence of these factors was made possible through an extended on-site case study. Methodology for this based upon participant observation coupled wth a wide and flexible range of techniques is described. Evidence is presented about many aspects of the innovation process from a number of different levels and perspectives: the attempt is to demonstrate the extent to which variation due to contingent influences takes place. It is argued that problems identified all relate to the issue of integration. This theme is also developed from an analytical viewoint and it is suggested that organisational response to increases in complexity in the external environment will be to match them with internal complexity. Differentiation of this kind will require extensive and flexible integration, especially in those inherently uncertain areas associated with innovation. Whilst traditionally a function of management, it is argued that integration needs have increased to the point where a new specialism is required. The concept of integration specialist is developed from this analysis and attempts at simple integrative change during the research are described. Finally a strategy for integration - or rather for building in integrative capability - ln the organisation studied is described.

The tribology of laminated magnetic recording heads

This thesis investigates the mechanisms that lead to pole tip recession (PTR) in laminated magnetic recording heads (also known as "sandwich heads"). These heads provide a platform for the utilisation of advanced soft magnetic thin films in practical recording heads suitable for high frequency helical scan tape recording systems. PTR results from a differential wear of the magnetic pole piece from the tape-bearing surface of the head. It results in a spacing loss of the playback or read signal of 54.6dB per recording wavelength separation of the poles from the tape. PTR depends on the material combination used in the head, on the tape type and the climate - temperature and relative humidity (r.h.). Five head materials were studied: two non-magnetic substrate materials- sintered multi granular CaTi03 and composite CaTi03/ZrTi04/Ti02 and three soft magnetic materials- amorphous CoNbZr, and nanocrystalline FeNbSiN and FeTaN. Single material dummy heads were constructed and their wear rates measured when cycling them in a Hi-8 camcorder against commercially available metal particulate (MP) and metal evaporated (ME) tapes in three different climates: 25°C/20%r.h., 25°C/80%r.h. and 40°C/80%r.h. X-ray photoelectron spectroscopy (XPS) was used to examine changes the head surface chemistry. Atomic force microscopy (AFM) was used to examine changes in head and tape surface topography. PTR versus cycling time of laminated heads of CaTi03/ZrTiO4/Ti02 and FeTaN construction was measured using AFM. The principal wear mechanism observed for all head materials was microabrasion caused by the mating body - the tape surface. The variation in wear rate with climate and tape type was due to a variation in severity in this mechanism, except for tape cycling at 40°C in which gross damage was observed to be occurring to the head surface. Two subsidiary wear mechanisms were found: third body scratching (all materials) and grain pullout (both ceramics and FeNbSiN). No chemical wear was observed, though tribochemical reactions were observed on the metal head surfaces. PTR was found to be caused by two mechanisms - the first differential microabrasion of the metal and substrate materials and which was characterised by a low (~10nm) equilibrium value. The second was by deep ploughing by third body debris particles, thought mainly to be grain pullout particles. This level of PTR caused by this mechanism was often more severe, and of a non-equilibrium nature. It was observed more for ME tape, especially at 40°C/80%r.h. and 25°c/20%r.h. Two other phenomena on the laminated head pole piece were observed and commented upon: staining and ripple texturing.

A theoretical and experimental analysis of the no-load air gap magnetic field in squirrel cage induction machines

In induction machines the tooth frequency losses due to permeance variation constitute a signif'icant, portion of the total loss. In order to predict and estimate these losses it, is essential to obtain a clear understanding of the no-load distribution of the air gap magnetic field and the magnitude of flux pulsation in both stator and rotor teeth. The existing theories and methods by which the air gap permeance variation in a doubly slotted structure is calculated are either empirical or restricted. The main objective of this thesis is to obtain a detailed analysis of the no-load air gap magnetic field distribution and the effect of air gap geometry on the magnitude and waveform of the tooth flux pulsation. In this thesis a detaiiled theoretical and experimental analysis of flux distribution not only leads to a better understanding of the distribution of no-load losses but also provides theoretical analysis for calculating the losses with greater accuracy

Creep and fatigue of a single crystal nickel base superalloy

The introduction of single crystal casting techniques has led to the development of existing nickel-base superalloys to produce materials with optimum mechanical properties in the single crystal condition. As single crystals are known to be anisotropic, a study is needed to determine the general mechanical properties of these materials, and determine the effects of crystal orientation upon them. A study has been carried out to identify the effect of orientation and temperature on the creep and fatigue properties of a development single crystal superalloy, SRR 99. Creep testing and crystal rotation experiments have been made on SRR 99 and an earlier development alloy, SRR 9. Fatigue experiments at elevated temperatures have been carried out on both notched and un-notched specimens of alloy SRR 99. To aid in this analysis, several analytical techniques have been employed including Laue x-ray orientation analysis, measurement of strain by photographic methods and microstructural examination. Crystal rotation experiments have indicated that shear of 1 precipitates by lbrace111rbrace< 112> slip systems is operative during primary creep deformation at temperatures of 750oC and 850oC. The effect of orientation variation obtained by standard casting practices was not found to be significant. Creep rupture was found to be associated with multiple crack initiation from micropores. Fatigue crack initiation in un-notched specimens was found to be related to microporosity and microstructural defects. Failure was predominantly by crystallographic crack growth on lbrace111rbrace planes. The use of linear elastic fracture mechanics to describe fatigue crack propagation in alloy SRR 99 was found to be acceptable at temperatures up to 850oC. Variation of temperature, frequency and crystal orientation was found to have only moderate effect upon crack propagation rates.

Individual differences in the sensory perception of fats

Excessive consumption of dietary fat is acknowledged to be a widespread problem linked to a range of medical conditions. Despite this, little is known about the specific sensory appeal held by fats and no previous published research exists concerning human perception of non-textural taste qualities in fats. This research aimed to address whether a taste component can be found in sensory perception of pure fats. It also examined whether individual differences existed in human taste responses to fat, using both aggregated data analysis methods and multidimensional scaling. Results indicated that individuals were able to detect both the primary taste qualities of sweet, salty, sour and bitter in pure processed oils and reliably ascribe their own individually-generated taste labels, suggested that a taste component may be present in human responses to fat. Individual variation appeared to exist, both in the perception of given taste qualities and in perceived intensity and preferences. A number of factors were examined in relation to such individual differences in taste perception, including age, gender, genetic sensitivity to 6-n-propylthiouracil, body mass, dietary preferences and intake, dieting behaviours and restraint. Results revealed that, to varying extents, gender, age, sensitivity to 6-n-propylthiouracil, dietary preferences, habitual dietary intake and restraint all appeared to be related to individual variation in taste responses to fat. However, in general, these differences appeared to exist in the form of differing preferences and levels of intensity with which taste qualities detected in fat were perceived, as opposed to the perception of specific taste qualities being associated with given traits or states. Equally, each of these factors appeared to exert only a limited influence upon variation in sensory responses and thus the potential for using taste responses to fats as a marker for issues such as over-consumption, obesity or eating disorder is at present limited.

Experimental and theoretical studies on a distributed temperature sensor based on Brillouin scattering

Results are reported from recent research on the use of the Brillouin gain/loss mechanism for distributed sensing. A theoretical model of the interaction of the pulsed and CW beams is described and compared with experiments. Results from a system with a 51 km sensing length are presented. We finally investigate issues related to the variation within the sensing fiber of the polarizations of the two beams.