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Biomedical subjects

M K Walsh

Publications and source records attributed to M K Walsh.

At least 19 recordsLinked to original sources

DAYCENT national-scale simulations of nitrous oxide emissions from cropped soils in the United States.

Until recently, Intergovernmental Panel on Climate Change (IPCC) emission factor methodology, based on simple empirical relationships, has been used to estimate carbon (C) and nitrogen (N) fluxes for regional and national inventories. However, the 2005 USEPA greenhouse gas inventory includes estimates of N2O emissions from cultivated soils derived from simulations using DAYCENT, a process-based biogeochemical model. DAYCENT simulated major U.S. crops at county-level resolution and IPCC emission factor methodology was used to estimate emissions for the approximately 14% of cropped land not simulated by DAYCENT. The methodology used to combine DAYCENT simulations and IPCC methodology to estimate direct and indirect N2O emissions is described in detail. Nitrous oxide emissions from simulations of presettlement native vegetation were subtracted from cropped soil N2O to isolate anthropogenic emissions. Meteorological data required to drive DAYCENT were acquired from DAYMET, an algorithm that uses weather station data and accounts for topography to predict daily temperature and precipitation at 1-km2 resolution. Soils data were acquired from the State Soil Geographic Database (STATSGO). Weather data and dominant soil texture class that lie closest to the geographical center of the largest cluster of cropped land in each county were used to drive DAYCENT. Land management information was implemented at the agricultural-economic region level, as defined by the Agricultural Sector Model. Maps of model-simulated county-level crop yields were compared with yields estimated by the USDA for quality control. Combining results from DAYCENT simulations of major crops and IPCC methodology for remaining cropland yielded estimates of approximately 109 and approximately 70 Tg CO2 equivalents for direct and indirect, respectively, mean annual anthropogenic N2O emissions for 1990-2003.

Agriculture↗

Performance of dairy cattle clones and evaluation of their milk composition.

Genetic and phenotypic performance of U.S. Holstein embryo-split and nuclear-transfer clones was documented for yield and fitness traits. For cows, mean genetic superiority based on pedigree was 186 kg of milk, 9 kg of fat, and 7 kg of protein for embryo-split clones and 165, 10, and 8 kg, respectively, for nuclear-transfer clones compared with the population for the same birth year; pedigree advantage for male clones generally was slightly greater. Estimates of genetic merit that considered a clone's own performance as well as pedigree merit were slightly lower for embryo-split cows than for their full siblings for yield but not for milk composition (fat and protein percentages), mastitis resistance (somatic cell score), longevity (productive life), or cow fertility (daughter pregnancy rate); no corresponding genetic differences were found for nuclear-transfer cows or for cloned bulls regardless of clone type. For bulls, estimated genetic merit based on daughter yield was more similar for clone pairs with apparent identical genotype than for pairs from the same biotechnology but nonidentical as confirmed by blood typing. Yield deviations were lower for clones than for their full siblings. Milk composition (total solids, fat, fatty acid profile, lactose, and protein) also was compared for nuclear-transfer clones (Brown Swiss, Holstein, and Holstein-Jersey cross) with non-cloned cows and literature values; no differences were found for gross chemical composition of milk. No obvious differences were evident between cloned and non-cloned animals or for the milk that they produced.

Animals↗

Risk-adjusted capitation funding models for chronic disease in Australia: alternatives to casemix funding.

Under Australian casemix funding arrangements that use Diagnosis-Related Groups (DRGs) the average price is policy based, not benchmarked. Cost weights are too low for State-wide chronic disease services. Risk-adjusted Capitation Funding Models (RACFM) are feasible alternatives. A RACFM was developed for public patients with cystic fibrosis treated by an Australian Health Maintenance Organization (AHMO). Adverse selection is of limited concern since patients pay solidarity contributions via Medicare levy with no premium contributions to the AHMO. Sponsors paying premium subsidies are the State of Victoria and the Federal Government. Cost per patient is the dependent variable in the multiple regression. Data on DRG 173 (cystic fibrosis) patients were assessed for heteroskedasticity, multicollinearity, structural stability and functional form. Stepwise linear regression excluded non-significant variables. Significant variables were 'emergency' (1276.9), 'outlier' (6377.1), 'complexity' (3043.5), 'procedures' (317.4) and the constant (4492.7) (R(2)=0.21, SE=3598.3, F=14.39, Prob<0.0001. Regression coefficients represent the additional per patient costs summed to the base payment (constant). The model explained 21% of the variance in cost per patient. The payment rate is adjusted by a best practice annual admission rate per patient. The model is a blended RACFM for in-patient, out-patient, Hospital In The Home, Fee-For-Service Federal payments for drugs and medical services; lump sum lung transplant payments and risk sharing through cost (loss) outlier payments. State and Federally funded home and palliative services are 'carved out'. The model, which has national application via Coordinated Care Trials and by Australian States for RACFMs may be instructive for Germany, which plans to use Australian DRGs for casemix funding. The capitation alternative for chronic disease can improve equity, allocative efficiency and distributional justice. The use of Diagnostic Cost Groups (DCGs) is a promising alternative classification system for capitation arrangements.

Journal Article↗

Asynchronous synapse elimination in neonatal motor units: studies using GFP transgenic mice.

In developing muscle, synapse elimination reduces the number of motor axons that innervate each postsynaptic cell. This loss of connections is thought to be a consequence of axon branch trimming. However, branch retraction has not been observed directly, and many questions remain, such as: do all motor axons retract branches, are eliminated branches withdrawn synchronously, and are withdrawing branches localized to particular regions? To address these questions, we used transgenic mice that express fluorescent proteins in small subsets of motor axons, providing a unique opportunity to reconstruct complete axonal arbors and identify all the postsynaptic targets. We found that, during early postnatal development, each motor axon loses terminal branches, but retracting branches withdraw asynchronously and without obvious spatial bias, suggesting that local interactions at each neuromuscular junction regulate synapse elimination.

Aging↗

Optimizing the immobilization of single-stranded DNA onto glass beads.

The attachment of single-stranded DNA to a solid support has many biotechnology and molecular biology applications. This paper compares different immobilization chemistries to covalently link single-stranded DNA (20 base pairs), oligo(1), onto glass beads via a 5'-amino terminal end. Immobilization methods included a one-step 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and a two-step EDC reaction to succinylated and PEG-modified glass beads. The third method used 1,4-phenylene diisothiocyanate to immobilize oligo(1) to aminopropyl glass beads. The influence of coupling buffer, oligo(1) concentration, and EDC concentration was also investigated. The one-step EDC-mediated procedure with succinylated or PEG-modified beads in 0.1 M MES buffer, pH 4.5, resulted in the highest immobilization efficiency, 82-89%. EDC concentrations greater than 50 mM and oligo(1) concentrations of 3 microg/g bead were required for effective immobilization. A complementary oligonucleotide, oligo(2), was able to hybridize to the immobilized oligo(1) with a 58% efficiency. This oligonucleotide was subsequently released at 70 degrees C. The relationship between the surface density of oligo(1) and the hybridization efficiency of the complementary oligonucleotide is described.

DNA, Single-Stranded↗

Rapid fractionation of bovine transferrin using immobilized gangliosides.

Bovine transferrin (BTF) was fractionated from bovine whey using ganglioside affinity chromatography. After loading the immobilized matrix with a 2% whey solution, the matrix was washed with sodium acetate buffer at pH 4 containing 1 M NaCl before elution of BTF with sodium phosphate buffers at pH 7. Concanavalin-A affinity and ion exchange chromatography were used for further purification. The ganglioside column showed a 74.2% BTF recovery from whey and BTF was enriched to 61% purity with ion exchange chromatography. Bovine transferrin was identified by SDS-PAGE and western analysis. The Concanavalin-A affinity and ion exchange chromatography steps enriched BTF in the samples and removed other whey proteins from ganglioside purified fractions. These results indicate that immobilized ganglioside can be used to fractionate BTF from bovine whey. Our novel ganglioside affinity chromatography is rapid and efficient for the fractionation of BTF from whey.

Animals↗

Affinity enrichment of bovine lactoferrin in whey.

Bovine lactoferrin was enriched in various whey samples by affinity chromatography using immobilized gangliosides. Bovine gangliosides were isolated from fresh buttermilk using a combination of ultrafiltration and organic extraction. Isolated gangliosides were covalently immobilized onto controlled-pore glass beads. The immobilized matrix contained 66 micrograms of gangliosides per gram of beads. After loading the matrix with reconstituted whey protein isolate (WPI) or whey protein concentrate (WPC), the matrix was washed with sodium phosphate buffer (pH 7) followed by sodium acetate buffer (pH 4) before elution of lactoferrin with 1 M NaCl in sodium acetate buffer. From the intensities of the protein bands in SDS-PAGE, lactoferrin constituted a minimum of 40% of the total protein in the salt eluted sample. WPI, pretrated by heating and ultrafiltration, showed the highest lactoferrin purity among protein sources, while WPI (10% wt/vol) showed the highest recovery. These results show that immobilized gangliosides can be used to enrich the lactoferrin content of whey.

Animals↗

Solid-phase capture of proteins, spores, and bacteria.

Current methods for the detection of pathogens in food and water samples generally require a preenrichment step that allows selective enrichment of the test organism. The objective of this research was to eliminate an enrichment step to allow detection of bacteria directly in food and water samples in 30 min. A high-flow-rate, fluidized bed to capture and concentrate large (bacteria and spores) and small (protein) molecules was developed. This format, ImmunoFlow, is volume independent and uses large beads (greater than 3 mm in diameter) when capturing bacteria to prevent sample clogging when testing food samples. Detection of bound targets was done using existing enzyme-linked immunosorbent assay (ELISA) protocols. Four antibodies (anti-Escherichia coli O157:H7, -Bacillus globigii, -bovine serum albumin [BSA], and -ovalbumin [OVA]) were covalently coupled to various glass and ceramic beads. Very small amounts of BSA (<1 ng) and OVA (0.2 to 4.0 microg) were detected. Various industrial and environmental samples were used to observe the effect of the sample composition on the capture of anti-B. globigii and anti-E. coli O157:H7 modified beads. The lower limit of detection for both E. coli O157:H7 and B. globigii was 1 spore/cell independent of the sample size. The activity of anti-B. globigii modified beads declined after 3 days. Anti-E. coli O157:H7 modified beads declined in their capture ability after 2 days in various storage buffers. Storage temperature (4 and 25 degrees C) did not influence the stability. The ImmunoFlow technology is capable of capturing bacteria and spores directly from samples, with subsequent detection in an ELISA format in 30 min.

Antibodies, Monoclonal↗

Interinstitutional variation in glycohemoglobin monitoring and glycemic control of diabetic patients.

OBJECTIVES: To compare how frequently institutions monitor glycohemoglobin in diabetic patients, the level of glycemic control achieved and to identify institutional factors associated with higher rates of monitoring and lower glycohemoglobin levels. METHODS: A total of 212 institutions retrospectively abstracted laboratory and outpatient records of up to 30 diabetic patients who had initial glycohemoglobin monitoring performed in their laboratories. Data from a cohort of 5586 diabetic patients and 17 365 assays were analyzed. RESULTS: Overall, 31.3% of patients underwent glycohemoglobin monitoring at least quarterly, the frequency recommended by the American Diabetes Association (ADA) to stabilize patients at target hemoglobin A(1c) (HbA(1c)) levels. A total of 64.9% of patients were monitored at least semiannually, the ADA recommendation for patients with stable diabetes in glycemic control (final HbA(1c) level <7%). When we compared the top and bottom deciles of the 212 institutions, there was more than an eightfold difference in the proportion of patients monitored at least quarterly and more than a twofold difference in the proportion of patients monitored at least semiannually. Glycemic control was assessed by examining the value of the last glycohemoglobin determination on record after at least 8 months of management. For all 5586 diabetic patients, the median value of the last HbA(1c) assay was 7.4%. Comparing the top and bottom deciles, there was almost a fourfold difference among institutions in the proportion of diabetic patients in glycemic control. The use of reminders to order glycohemoglobin monitoring was associated with higher rates of semiannual monitoring (P <.05) and tighter glycemic control (P <.05). In addition, patients who were monitored more frequently experienced glycohemoglobin reductions of greater magnitude (P <.001). The presence of diabetes clinics and the use of rapid methods for testing glycohemoglobin were not associated with monitoring frequency or glycohemoglobin levels. CONCLUSIONS: There is wide interinstitutional variation in the frequency with which diabetic patients are monitored and the level of glycemic control achieved. The use of prompting systems to remind providers to order glycohemoglobin monitoring was associated with more frequent monitoring and superior glycemic control.

Blood Glucose↗

Solitary blood cultures: a College of American Pathologists Q-probes study of 132,778 blood culture sets in 333 small hospitals.

OBJECTIVE: To determine the frequency with which solitary blood culture samples were submitted to laboratories serving small hospitals and to ascertain whether certain hospital practices relating to the performance of blood cultures were associated with lower solitary blood culture rates (SBCRs). DESIGN: Participants in the College of American Pathologists Q-Probes laboratory quality improvement program collected data prospectively on the numbers of solitary blood culture sets from adult patients submitted to their laboratories and answered questions about their institutions' practice characteristics relating to the collection of blood culture specimens. SETTING AND PARTICIPANTS: Three hundred thirty-three public and private institutions with a median occupied bed size of 57. Participants were located in the United States (n = 329), Canada (n = 3), and Australia (n = 1). MAIN OUTCOME MEASURE: The solitary blood culture rate was defined as the number of instances in which only 1 blood culture venipuncture was performed on an individual patient during a 24-hour period divided by the total number of blood culture venipunctures that were performed during the study period. RESULTS: Participants submitted data on 132 778 adult patient blood culture sets. The SBCRs were 3.4% or less in the top-performing 10% of participating institutions (90th percentile and above), 12.7% in the midrange of participating institutions (50th percentile), and 42.5% or more in the bottom-performing 10% of participating institutions (10th percentile and below). In half the participating institutions, the SBCRs for inpatients were 8.3% or less and for outpatients, 22% or less. Solitary blood culture rates were lower for institutions in which phlebotomists rather than nonphlebotomists routinely collected blood culture specimens, in which internal policies required drawing at least 2 blood culture sets, in which hospital personnel contacted clinicians when their laboratories received requests for solitary blood culture sets, and in which quality control programs monitored SBCRs routinely. CONCLUSIONS: Hospitals can achieve SBCRs under 5%. Those hospitals with particularly high SBCRs may lower their rates by altering certain institutional practices.

Adult↗

Influence of a poly-ethylene glycol spacer on antigen capture by immobilized antibodies.

The use of spacers to distance an immobilized antibody from the surface of a support matrix introduces flexibility, which can reduce steric interferences between antibodies leading to a higher antigen capture efficiency. In this paper we investigated the use of a spacer molecule, poly-ethylene glycol (PEG), between the matrix surface and antibodies for the capture of Bacillus globigii, E. coli O157:H7, and ovalbumin. The antigen capture efficiency was determined using a surface ELISA method. Antibodies against the antigens were covalently immobilized either directly or via PEG to glass surfaces using a one-step EDC reaction. The amount of antibody immobilized was determined before blocking the nonspecific binding sites with bovine serum albumin. Antibodies immobilized via a PEG spacer showed a higher capture efficiency compared to direct immobilization, which was more pronounced with large antigens. Antibodies immobilized on glass supports were stable at 65 degrees C for at least 80 min, and the capture efficiency increased with heating at 65 degrees C for 20 min.

Animals↗

Use of amino acid analysis for estimating the individual concentrations of proteins in mixtures.

The concentrations of five individual proteins in a mixture were determined from one amino acid analysis of the mixture by solving for each protein using simultaneous equations. Dried casein and whey were separated into five individual protein components using reversed-phase HPLC. Individual proteins were collected and analyzed for amino acid composition. These data were used as standards. Mixtures of purified proteins were analyzed for total amino acid composition and the concentrations of individual proteins in the mixtures were determined by solving simultaneous equations based on the amino acid analysis composition of the standards.

Amino Acids↗

Characterization and expression of the pseudomonas putida bacterioferritin alpha subunit gene.

The root-colonizing pseudomonad Pseudomonas putida (Pp) appears to produce two subunits, alpha and beta, of the iron-binding protein, bacterioferritin. A gene encoding the alpha-bacterioferritin subunit was located adjacent to the major catalase in Pp. The deduced protein sequence of the Pp bfralpha gene had a very high identity with other alpha-subunits, possessing conserved amino acids responsible for ferroxidase activity. The gene also lacked a deduced methionine at residue 52, associated with heme binding in beta-subunits. An antibody generated toward the Escherichia coli (E. coli) multifunctional single subunit bacterioferritin recognized two proteins in the Pp extract, a 22 kDa protein likely to be a beta-subunit and, to a lesser extent, a 23 kDa band. The 23 kDa band was absent in a Pp mutant in which the bfralpha gene was disrupted. Loss of alpha-bacterioferritin stimulated production of fluorescent siderophore. Growth on media and on root surfaces was not impaired by deletion of the alpha-bacterioferritin. Transcription of bfralpha was independent of the catalase gene and was dependent on iron. The transcript levels from bfralpha decreased in iron deficiency experienced during stationary-phase or upon treatment during growth with an iron chelator.

Bacterial Proteins↗

Forecasting hospital expenditure in Victoria: lessons from Europe and Canada.

This paper specifies an econometric model to forecast State government expenditure on recognised public hospitals in Victoria. The OECD's recent cross-country econometric work exploring factors affecting health spending was instructive. The model found that Victorian Gross State Product, population aged under 4 years, the mix of public and private patients in public hospitals, introduction of casemix funding and funding cuts, the proportion of public beds to total beds in Victoria and technology significantly impacted on expenditure. The model may have application internationally for forecasting health costs, particularly in short and medium-term budgetary cycles.

Adolescent↗

A new ambulatory classification and funding model for radiation oncology: non-admitted patients in Victorian hospitals.

The Victorian Department of Human Services has developed a classification and funding model for non-admitted radiation oncology patients. Agencies were previously funded on an historical cost input basis. For 1996-97, payments were made according to the new Non-admitted Radiation Oncology Classification System and include four key components. Fixed grants are based on Weighted Radiation Therapy Services targets for megavoltage courses, planning procedures (dosimetry and simulation) and consultations. The additional throughput pool covers additional Weighted Radiation Therapy Services once targets are reached, with access conditional on the utilisation of a minimum number of megavoltage fields by each hospital. Block grants cover specialised treatments, such as brachytherapy, allied health payments and other support services. Compensation grants were available to bring payments up to the level of the previous year. There is potential to provide incentives to promote best practice in Australia through linking appropriate practice to funding models. Key Australian and international developments should be monitored, including economic evaluation studies, classification and funding models, and the deliberations of the American College of Radiology, the American Society for Therapeutic Radiology and Oncology, the Trans-Tasman Radiation Oncology Group and the Council of Oncology Societies of Australia. National impact on clinical practice guidelines in Australia can be achieved through the Quality of Care and Health Outcomes Committee of the National Health and Medical Research Council.

Abstracting and Indexing↗

Fostering progress and development (in an anaesthetic department) at a time of cost containment.

The lessons learned in industry can be applied to anaesthetic departments. Progress and development are part of improving quality, and evidence is emerging that improved quality can reduce costs. Improving quality requires anaesthetic departments to develop a clear customer focus. Education, training, research and quality improvement are essential components of a quality anaesthetic department. Some of the cost reductions are achieved by development of partnerships with customers and suppliers. The emphasis is likely to shift from cost reduction to quality improvement and anaesthetic departments should anticipate this change.

Journal Article↗

Fish consumption and the 30-year risk of fatal myocardial infarction.

BACKGROUND: Epidemiologic data on the possible benefit of eating fish to reduce the risk of coronary heart disease have been inconsistent. We used data from the Chicago Western Electric Study to examine the relation between base-line fish consumption and the 30-year risk of death from coronary heart disease. METHODS: The study participants were 1822 men who were 40 to 55 years old and free of cardiovascular disease at base line. Fish consumption, as determined from a detailed dietary history, was stratified (0, 1 to 17, 18 to 34, and > or = 35 g per day). Mortality from coronary heart disease, ascertained from death certificates, was classified as death from myocardial infarction (sudden or nonsudden) or death from other coronary causes. RESULTS: During 47,153 person-years of follow-up, there were 430 deaths from coronary heart disease; 293 were due to myocardial infarctions (196 were sudden, 94 were nonsudden, and 3 were not classifiable). Cox proportional-hazards regression showed that for men who consumed 35 g or more of fish daily as compared with those who consumed none, the relative risks of death from coronary heart disease and from sudden or nonsudden myocardial infarction were 0.62 (95 percent confidence interval, 0.40 to 0.94) and 0.56 (95 percent confidence interval, 0.33 to 0.93), respectively, with a graded relation between the relative risks and the strata of fish consumption (P for trend = 0.04 and 0.02, respectively). These findings were accounted for by the relation of fish consumption to nonsudden death from myocardial infarction (relative risk, 0.33; 95 percent confidence interval, 0.12 to 0.91; P for trend= 0.007). CONCLUSIONS: These data show an inverse association between fish consumption and death from coronary heart disease, especially nonsudden death from myocardial infarction.

Adult↗