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An integrated data system for the retail pharmaceutical service.

A machine has been designed for use in a retail pharmacy which counts tablets and records the count beside a machine-read bar code indicating the identity of the drug. Such data could be used for retail stock control and passed on for use by wholesaler, manufacturer and prescription pricing bureau for their routine processes. Thus present processes would be greatly simplified and some other advantages gained.

Drug and Narcotic Control↗

Determination of total and free plasma carnitine concentrations on the Dade Behring Dimension RxL: integrated chemistry system.

BACKGROUND: L-carnitine is a naturally occurring quaternary ammonium compound present in all mammalian species. Its major function is to facilitate the passage of long-chain fatty acids through the mitochondrial membrane for subsequent beta-oxidation and ketone synthesis. Clinical interest in carnitine disorders relates particularly to possible deficiency states that may result in a phenotypic spectrum that includes cardiomyopathy, skeletal myopathy, hypoglycemia and hyperammonemia. The objective of this study was to develop a method on the Dade Behring Dimension RxL analyzer for measuring free and total carnitine levels in plasma. METHODS: Plasma samples were deproteinized by ultrafiltration to remove interference by endogenous thiols. Filtrates were measured directly on the RxL for free carnitine or after alkaline hydrolysis for total carnitine by an endpoint enzymatic assay that uses carnitine acetyltransferase. RESULTS: Within-run imprecision was <5% at high and low levels for both free and total carnitine while between-day imprecision was <15%. Recovery of free carnitine from spiked plasma >90%. The method was linear between 5.0 and 150.0 micromol/l and the limit of quantification was 5.0 micromol/l. Comparison of our method with another automated procedure developed on the Hitachi 917 system using Deming regression analysis resulted in the following equations: Dimension=1.034(Hitachi)-7.44 for total carnitine (r=0.955) and Dimension=0.805(Hitachi)+1.96 for free carnitine (r=0.951), respectively. CONCLUSIONS: Our method is suitable for analyzer platforms where the level of imprecision is lower and the throughput is higher than manual methods. It also avoids the use of radioisotopes and is appropriate in labs where access to reference methods such as tandem mass spectrometry and HPLC is limited or unavailable.

Carnitine↗

Pharmacogenomics and cardiovascular drugs: need for integrated biological system with phenotypes and proteomic markers.

Personalized medicine is based on a better knowledge of biological variability, considering the important part due to genetics. When trying to identify involved genes and their products in differential cardiovascular drug responses, a five-step strategy is to be followed: 1) Pharmacokinetic-related genes and phenotypes (2) Pharmacodynamic targets, genes and products (3) Cardiovascular diseases and risks depending on specific or large metabolic cycles (4) Physiological variations of previously identified genes and proteins (5) Environment influences on them. After summarizing the most well-known genes involved in drug metabolism, we will take as example of drugs, the statins, considered as very important drugs from a Public-Health standpoint, but also for economical reasons. These drugs respond differently in human depending on multiple polymorphisms. We will give examples with common ApoE polymorphisms influencing the hypolipemic effects of statins. These drugs also have pleiotropic effects and decrease inflammatory markers. This illustrates the need to separate clinical diseases phenotypes in specific metabolic pathways, which could propose other classifications, of diseases and related genes. Hypertension is also a good example of clinical phenotype which should be followed after various therapeutic approaches by genes polymorphisms and proteins markers. Gene products are under clear environmental expression variations such as age, body mass index and obesity, alcohol, tobacco and dietary interventions which are the first therapeutical actions taken in cardiovascular diseases. But at each of the five steps, within a pharmacoproteomic strategy, we also need to use available information from peptides, proteins and metabolites, which usually are the gene products. A profiling approach, i.e., dealing with genomics, but now also with proteomics, is to be used. In conclusion, the profiling, as well as the large amount of data, will more than before render necessary an organized interpretation of DNA, RNA as well as proteins variations, both at individual and population level.

Cardiovascular Agents↗

A genome-wide expression profile and system-level integration of nuclear factor kappa B regulated genes reveals fundamental metabolic adaptations during cell growth and survival.

A murine lung alveolar carcinoma cell line (WT-Line 1) and its equally tumorigenic but non-malignant derivative transduced with a dominant negative inhibitor of NF-kappaB (mI-kappaB-Line 1), were profiled on the Affymetrix 19000 gene array platform. Two differentially expressed gene clusters were identified and integrated into a functional model. The downregulation of anti-oxidant defenses, in mI-kappaB-Line 1 cells, correlates with high levels of reactive oxygen species (ROS) and ROS damage to cellular macromolecules while the upregulation of metabolic nuclear receptors correlates with an adaptive/survival response, which involves a shift in energy metabolism toward beta-oxidative respiration. Accordingly, mI-kappaB-Line 1 cells are markedly sensitized to pharmacologic inhibition of beta-oxidative respiration. These findings are indicative of compensatory changes that could undermine anti-cancer therapies targeting NF-kappaB.

Adaptation, Physiological↗

The importance of quality of survival as an outcome measure for an integrated trauma system.

Risk-adjusted survival rates have been the principle mode of comparison between trauma systems. In mature trauma systems, it is possible that there will be further improvements in survival but these are likely to be small. In the future, the largest gains will come from quality of life and improved function of the survivors. The issues related to measuring quality of survival for trauma systems are reviewed, including feasibility, ethical considerations, risk adjustment of outcomes of survivors, and challenges for selection of instruments and administration. In addition, the preliminary experiences of measuring outcomes in survivors through the Victorian State Trauma Registry are discussed. Although function and quality of life have been identified as important factors to measure in trauma populations, a standardised protocol has not been established. The experience in Victoria suggests that monitoring of population-based outcomes in survivors is feasible and may create the basis for benchmarking the level of morbidity in survivors.

Australia↗

Real-time transmission of pediatric echocardiograms using a single ISDN line.

We tested the adequacy of a videoconferencing system using a single integrated systems digital network (ISDN) line (128 kilobits per second) for the remote diagnosis of children with suspected congenital heart disease (CHD). Real-time echocardiogram interpretation was compared to subsequent videotape review in 401 studies with concordance in 383 (95.5%) studies. A new diagnosis of CHD was made in 98 studies. Immediate patient transfer was arranged based upon a real-time diagnosis in five studies. In 300 studies, a normal diagnosis obviated further evaluation. A single ISDN line is adequate for transmission of pediatric echocardiograms and it allows for remote management of patients with CHD.

Adolescent↗

An integrated vector system for the eukaryotic expression of antibodies or their fragments after selection from phage display libraries.

Phage display is now an established method to select antibody fragments specific for a wide range of diverse antigens. In particular, isolation of human monoclonal antibodies has become a reality and for most purposes bacterial expression of the selected recombinant antibody fragments is sufficient. However, there are some cases where the expression of complete human immunoglobulin in mammalian cells is, if not essential, at least desirable. For this reason we have designed and constructed a set of mammalian expression vectors which permit facile and rapid cloning of antibody genes for both transient and stable expression in mammalian cells. Immunoglobulin genes may be cloned into these expression vectors as V regions or as Fabs for expression as either complete antibodies or as Fab fragments, using restriction sites which are rare in human V genes. All the important elements in the vectors--promoter, leader sequence, constant domains and selectable markers--are flanked by unique restriction sites, allowing simple substitution of elements. The vectors have been evaluated using the variable regions from the neutralizing anti-nerve growth factor (NGF) antibody, alphaD11, and the V regions from 2E10, a scFv selected from a scFv phagemid library.

Animals↗

[Methologic contribution to blood transfusion materials surveillance].

To reduce seriousness and frequency of iatrogenic risk implies prevention policies and efficient operational systems for vigilance. This risk management implies definition of precise organizations and procedures able to locate and to notify quickly undesirable events. This is the case about single use medical devices (SUMD) used in blood transfusion. This article is a contribution to the organisation of the implemented material vigilance in blood transfusion, collectively carried out with actors concerned (users, manufacturers, National Commission for Material Vigilance). It presents a lot of tools and methods to favour practices harmonization, as well as preventive a curative (specifications before purchase, main part of the quality contract between customer and supplier; internal control plan; index for medical device used in transfusion; illustrated glossaries for three main families of medical devices; index about symptomatic events; definitions of seriousness levels with their operational consequences; methods to manage a single use medical device judged as defective; tool for the review of incidents according to reference and batch). Then, the management of incidents about SUMD is presented within a material vigilance system integrated into the quality system of the institution, for user as for manufacturer. This is done in a chronological order with successively description of the incident, the assessment of the impact, the management of the associated risk, the periodical review of incidents and management of matters in dispute.

Algorithms↗

Accuracy of alternative representations for integrated biochemical systems.

The Michaelis-Menten formalism often provides appropriate representations of individual enzyme-catalyzed reactions in vitro but is not well suited for the mathematical analysis of complex biochemical networks. Mathematically tractable alternatives are the linear formalism and the power-law formalism. Within the power-law formalism there are alternative ways to represent biochemical processes, depending upon the degree to which fluxes and concentrations are aggregated. Two of the most relevant variants for dealing with biochemical pathways are treated in this paper. In one variant, aggregation leads to a rate law for each enzyme-catalyzed reaction, which is then represented by a power-law function. In the other, aggregation produces a composite rate law for either net rate of increase or net rate of decrease of each system constituent; the composite rate laws are then represented by a power-law function. The first variant is the mathematical basis for a method of biochemical analysis called metabolic control, the latter for biochemical systems theory. We compare the accuracy of the linear and of the two power-law representations for networks of biochemical reactions governed by Michaelis-Menten and Hill kinetics. Michaelis-Menten kinetics are always represented more accurately by power-law than by linear functions. Hill kinetics are in most cases best modeled by power-law functions, but in some cases linear functions are best. Aggregation into composite rate laws for net increase or net decrease of each system constituent almost always improves the accuracy of the power-law representation. The improvement in accuracy is one of several factors that contribute to the wide range of validity of this power-law representation. Other contributing factors that are discussed include the nonlinear character of the power-law formalism, homeostatic regulatory mechanisms in living systems, and simplification of rate laws by regulatory mechanisms in vivo.

Biochemistry↗

Integrated nanoreactor systems: triggering the release and mixing of compounds inside single vesicles.

We present a method that allows the on-demand release and mixing of zepto- to femtoliter volumes of solutions in the interior of vesicular nanoreactors. The reactors comprise a nested system of lipid vesicles, part of which release their cargo in the interior of the others during a thermotropic phase transition. The performance of individual reactors immobilized on glass is characterized using confocal microscopy and a fluorescent dye that reports dilution during the release. The results confirm the predicted temperature-induced response and reveal a release transition width of 3 degrees C with a half time of approximately 1 min.

Lipid Bilayers↗