Population child health: understanding and addressing complex health needs.

Advances in paediatric care mean that more children with complex medical problems (heart disease, neurodevelopmental problems and so on) are surviving their early years. This has important implications for the design and delivery of healthcare given their extensive multidisciplinary requirements and susceptibility to poor outcomes when not optimally managed. Importantly, their medical needs must also be understood and addressed within the context of the child and family's life circumstances. There is growing recognition that many other factors contribute to a child's complex health needs (CHNs), for example, family problems, fragmentation of health and care provision, psychological difficulties or social issues.To facilitate proactive care for these patients, we must develop accurate ways to identify them. Whole Systems Integrated Care-an online platform that integrates routinely collected data from primary and secondary care-offers an example of how to do this. An algorithm applied to this data identifies children with CHNs from the entire patient population. When tested in a large inner-city GP practice, this analysis shows good concordance with clinical opinion and identifies complex children in the population to a much higher proportion than expected. Ongoing refinement of these data-driven processes will allow accurate quantification and identification of need in local populations, thus aiding the development of tailored services.

Discovery of xanthine oxidase inhibitors from a complex mixture using an online, restricted-access material coupled with column-switching liquid chromatography with a diode-array detection system.

To find potential lead compounds for antigout drug discovery, an automated online, restricted-access material coupled with column-switching liquid chromatography with a diode-array detection (RAM-LC-DAD) system was developed for screening of xanthine oxidase (XO) inhibitors and their affinity rankings in complex mixtures. The system was first evaluated by analyzing a mixture of six compounds with known inhibition of XO. Nonspecific binding to the denatured XO was investigated and used as the control for screening. Subsequently, the newly developed system was applied to screening of a natural product, Oroxylum indicum extract, and four compounds which could specifically interact with XO were found and identified as oroxin B, oroxin A, baicalin, and baicalein. The results were verified by a competitive binding test using the known competitive inhibitor allopurinol and were further validated by an inhibition assay in vitro. The online RAM-LC-DAD system developed was shown to be a simple and effective strategy for the rapid screening of bioactive compounds from a complex mixture.

Analysis of free and esterified sterols in edible oils by online reversed phase liquid chromatography-gas chromatography (RPLC-GC) using the through oven transfer adsorption desorption (TOTAD) interface.

An online reversed phase liquid chromatography-gas chromatography (RPLC-GC) method is proposed to quantify free, total and esterified sterols of edible oils. To determine free sterols the diluted oils are injected into the liquid chromatograph, where free sterols are separated from triglycerides and the sterol fraction is automatically transferred to the gas chromatograph to be analysed. To determine total sterols the samples were saponified with potassium hydroxide in ethanolic solution and the unsaponifiable fraction was extracted with diethyl ether. The extract was then analysed by RPLC-GC, avoiding the laborious thin layer chromatography step used in the Official European Union (EU) Method. The relative standard deviations (RSDs) from the absolute peak area varied from 7.6% to 15.8%. Limits of detection (LODs) were less than 8.5 mg/kg. No variability in retention time was observed. The method was applied to the determination of total sterols in edible oil samples and the results were compared with those obtained with the Official EU Method. Good agreement was found between both methods, except in the case of campesterol.

Gas chromatographic high-throughput screening techniques in catalysis.

Discovering highly efficient catalysts is of great scientific and economical interest. Advances in high-throughput assays in combination with sophisticated analytical techniques have increased the rapidity with which catalysts can be identified and optimized. Understanding how kinetics in the mechanism of catalysis is controlled by structural parameters is essential for a directed design of catalysts. To identify such rate-controlling elementary steps and to develop and refine models, comprehensive experimental kinetic data of a broad variety of substrates are necessary. In the present article concepts of high-throughput screening techniques in catalysis using gas chromatography are reviewed in a survey covering the period from 1998 to 2007. To cover also the origins of concepts and groundbreaking experiments in this research area milestones going back to 1950 are also reviewed. The first part of the review will focus on off-line gas chromatographic analysis, the second part on on-line gas chromatographic analysis covering sequential, parallelized and high-throughput multiplexing gas chromatography. The third part presents recent advances in the integration of chemical transformation and analysis in gas chromatography. The present review article describes the state-of-the-art, scope and limitations, and applications of these different high-throughput screening approaches.

HPLC micro-fractionation coupled to a cell-based assay for automated on-line primary screening of calcium antagonistic components in plant extracts.

High performance liquid chromatography (HPLC) micro-fractionation was successfully coupled to an automated 45Ca2+ uptake assay using GH4C1 cells for the separation of natural product extracts and for the primary detection of their calcium antagonistic components. The reliability of the procedure was first established with a reference solution consisting of pure compounds with a known effect on the Ca2+ uptake. No loss of activity was observed to occur after HPLC micro-fractionation. Extracts of Peucedanum palustre and Pinus sylvestris, showing high and no inhibition of Ca2+ uptake as total extracts, respectively, were analysed and the inhibitory activity of the P. palustre extract could be traced to two components, identified as columbianadin and isoimperatorin. As expected, no significant inhibition was observed with the micro-fractionated P. sylvestris samples. In summary, the procedure was found to be applicable for primary detection of calcium antagonistic components in complex matrices and to significantly reduce the time previously needed for bioactivity-guided isolation.

Electroencephalographic(EEG)-based communication: EEG control versus system performance in humans.

People can learn to control electroencephalographic (EEG) sensorimotor rhythm amplitude so as to move a cursor to select among choices on a computer screen. We explored the dependence of system performance on EEG control. Users moved the cursor to reach a target at one of four possible locations. EEG control was measured as the correlation (r(2)) between rhythm amplitude and target location. Performance was measured as accuracy (% of targets hit) and as information transfer rate (bits/trial). The relationship between EEG control and accuracy can be approximated by a linear function that is constant for all users. The results facilitate offline predictions of the effects on performance of using different EEG features or combinations of features to control cursor movement.

[Model construction for reperfusion of the isolated pig liver]. / Versuchsaufbau zur Reperfusion der isolierten Schweineleber.

In recent years thermal ablation of liver tumors as a minimally invasive method became a promising alternative to conventional strategies such as chemotherapy or resection of liver tissue. Thereby an electrode is placed inside the tumor delivering energy in the form of high frequency current into the target volume to achieve and maintain a tissue temperature between 60 and 100 degrees C. Cells exposed to this thermic stress undergo coagulation necrosis and are irreversibly damaged. To protect vital liver structures from heat, it is necessary to develop an online temperature monitoring system. An experimental setup perfusing isolated pig livers under physiological conditions with 0.9% NaCl solution was established to develop and evaluate the measuring technique.

On-line instrument confirmation: how can we check that our instruments are working?

As on-line instrumentation becomes increasingly important for operating modern wastewater treatment plants it is necessary to have diagnostic methods that can indicate sensor deviation at an early stage. Examples of on-line diagnosis methods used at the Rya WWTP (Göteborg, Sweden) are presented. These include on-line mass balance calculations and soft sensors, which make indirect parameter estimates. By comparing the estimates with the corresponding on-line sensors it is possible to make control decisions and to chose alternative strategies. These comparative methods are often useful for process monitoring and diagnosis as well. Some other soft sensor applications using exponential filters are also presented.

Setup and procedure for online identification of electrically stimulated muscle with Matlab Simulink.

This paper first describes a laboratory setup for biomechanical experiments that runs within the universal simulation environment Matlab Simulink. The overall system comprises a personal computer, two AMTI (Advanced Mechanical Technology, Inc., Watertown, MA 02472) force plates, Parotec force-sensor shoe insoles, Optotrak system for noncontact three-dimensional (3-D)-position measuring, and a computer-controlled four-channel electrical stimulator. Conceptually, the most important application is implementation of closed-loop electrical stimulation of intact and paralyzed subjects in the laboratory. Second, the system was tested in real-time muscle model identification procedure during a standing experiment. The plantarflexors of three nonimpaired subjects were excited with pseudorandom binary sequences (PRBSs) with small deviations around selected operating points. Electrically stimulated muscles were presented with a linear local dynamic block that was identified with a recursive least-square method (RARX). RARX block was designed with fundamental Matlab Simulink blocks that support real-time operation. Introduced was online estimation of model output, which offers a great manner of instant model validation. Two modes of operation with online validation were tested. In the first mode, the operating point for selected excitation level was identified online. In the second mode, the operating point was measured in preceding experiments. Both procedures resulted in satisfying second-order models that will be used in the adaptive controller design.

Clinical cerebral microdialysis: a methodological study.

OBJECT: Clinical microdialysis enables monitoring of the cerebral extracellular chemistry of neurosurgical patients. Introduction of the technique into different hospitals' neurosurgical units has resulted in variations in the method of application. There are several variables to be considered, including length of the catheter membrane, type of perfusion fluid, flow rate of perfusion fluid, and on-line compared with delayed analysis of samples. The objects of this study were as follows: 1) to determine the effects of varying catheter characteristics on substance concentration; 2) to determine the relative recovery and true extracellular concentration by varying the flow rate and extrapolating to zero flow; and 3) to compare substance concentration obtained using a bedside enzyme analyzer with that of off-line high-performance liquid chromatography (HPLC). METHODS: A specially designed bolt was used to conduct two adjacent microdialysis catheters into the frontal cortex of patients with head injury or poor-grade subarachnoid hemorrhage who were receiving ventilation. One reference catheter (10-mm membrane, perfused with Ringer's solution at 0.3 microl/minute) was constant for all studies. The other catheter was varied in terms of membrane length (10 mm or 30 mm), perfusion fluid (Ringer's solution or normal saline), and flow rate (0.1-1.5 microl/minute). The effect of freezing the samples on substance concentration was established by on-line analysis and then repeated analysis after storage at -70 degrees C for 3 months. Samples assayed with the bedside enzyme analyzer were reassessed using HPLC for the determination of glutamate concentrations. CONCLUSIONS: Two adjacent microdialysis catheters that were identical in membrane length, perfusion fluid, and flow rate showed equivalent results. Variations in perfusion fluid and freezing and thawing of samples did not result in differences in substance concentration. Catheter length had a significant impact on substance recovery. Variations in flow rate enabled the relative recovery to be calculated using a modification of the extrapolation-to-zero-flow method. The recovery was approximately 70% at 0.3 microl/minute and 30% at 1 microl/minute (10-mm membrane) for all analytes. Glutamate results obtained with the enzyme analyzer showed good correlation with those from HPLC.

Mathematical algorithm for qualitative and semiquantitative analysis of petroleum hydrocarbons in solid wastes using on-line gas chromatography.

Non-degradated mineral-oils like gasoline, solvent naphtha, diesel fuel, fuel and lubricating oils provide a characteristic fingerprint gas chromatogram. This visual classification, e.g. in solid wastes, is complicated due to the simultaneous presence of several mineral-oils. Therefore, a mathematical algorithm for the separation of gas chromatographic fingerprint of "single mixtures" of aliphatic hydrocarbons is developed. The technique is essential for analysis of time-overlapping "single mixtures" of petroleum hydrocarbons (so-called "complex mixtures") and it relies on the concentration-varying hydrocarbons during evaporation. It is possible to separate the data from the gas chromatogram of a "complex mixture" of hydrocarbons into the chromatograms of the pure "single mixtures" and to give their respective concentrations. A synthetic "complex mixture" of kerosene, diesel fuel and lubricating oil is used to illustrate the method.

Timing of EEG-based cursor control.

Recent studies show that humans can learn to control the amplitude of electroencephalography (EEG) activity in specific frequency bands over sensorimotor cortex and use it to move a cursor to a target on a computer screen. EEG-based communication could be a valuable new communication and control option for those with severe motor disabilities. Realization of this potential requires detailed knowledge of the characteristic features of EEG control. This study examined the course of EEG control after presentation of a target. At the beginning of each trial, a target appeared at the top or bottom edge of the subject's video screen and 1 sec later a cursor began to move vertically as a function of EEG amplitude in a specific frequency band. In well-trained subjects, this amplitude was high at the time the target appeared and then either remained high (i.e., for a top target) or fell rapidly (i.e., for a bottom target). Target-specific EEG amplitude control began 0.5 sec after the target appeared and appeared to wax and wane with a period of approximately 1 sec until the cursor reached the target (i.e., a hit) or the opposite edge of the screen (i.e., a miss). Accuracy was 90% or greater for each subject. Top-target errors usually occurred later in the trial because of failure to reach and/or maintain sufficiently high amplitude, whereas bottom-target errors usually occurred immediately because of failure to reduce an initially high amplitude quickly enough. The results suggest modifications that could improve performance. These include lengthening the intertrial period, shortening the delay between target appearance and cursor movement, and including time within the trial as a variable in the equation that translates EEG into cursor movement.

Fat content for nutritional labeling by supercritical fluid extraction and an on-line lipase catalyzed reaction.

A method using sequential supercritical fluid extraction (SFE) and enzymatic transesterification has been developed for the rapid determination of total nutritional fat content in meat samples. SFE conditions of 12.16 MPa and 50 degrees C were utilized to extract lipid species from the sample matrix. The enzymatic transesterification of the lipids by methanol was catalyzed by an immobilized lipase isolated from Candida antarctica. Conversion of the triglycerides to fatty acid methyl esters was monitored by supercritical fluid chromatography, while the fatty acid content of the extract was determined by capillary gas chromatography (GC). Total fat, saturated fat and monounsaturated fat contents were calculated from the GC data and compared to values from traditional extraction and lipid determination methods. Both off-line SFE and automated SFE followed by on-line GC analysis using two different instruments were utilized in this study. The enzymatic-based SFE method gave comparable results to the organic solvent extraction-based method followed by conventional BF3-catalyzed esterification.

Determination of polycyclic aromatic hydrocarbons in edible oils and fats by on-line donor-acceptor complex chromatography and high-performance liquid chromatography with fluorescence detection.

Various off-line methods for clean-up and sample enrichment are available for the analysis of polycyclic aromatic hydrocarbons (PAHs) in edible oils and fats. These methods consist of laborious and time consuming procedures. This study reports an on-line method using LC-LC coupling. After clean-up of the sample on a donor-acceptor complex chromatography (DACC) column the PAHs are transferred to and separated on an analytical HPLC column. Quantification is carried out with fluorescence detection. The DACC column clean-up is fast and is carried out during the HPLC run of the previous sample. Compared to the traditional methods this automated on-line method saves considerable time and significantly reduces the amount of solvent waste. The method uses common HPLC equipment and its performance has been evaluated.

Automated on-line gel permeation chromatography-gas chromatography for the determination of organophosphorus pesticides in olive oil.

An on-line combination of gel permeation chromatography and gas chromatography has been designed using either a laboratory-built or a commercially available LC-GC apparatus to determine organophosphorus pesticides in olive oil. Gel permeation chromatography was used for sample pretreatment, viz. to separate the low-molecular-mass pesticides from the higher-molecular-mass fat constituents of the oil. A mixture of n-decane and the azeotropic mixture of ethyl acetate and cyclohexane was found to give an adequate separation between the fat and the organophosphorus pesticides. The pesticide-containing fraction, monitored by a UV detector, was transferred on-line to the gas chromatograph using a loop-type interface. n-Decane (6%, v/v) was added to the eluent in order to widen the application range of the transfer technique towards more volatile pesticides. After solvent evaporation through the solvent vapour exit and subsequent GC separation, the compounds were selectively detected with a thermionic or a flame photometric detector. The set-up allowed the direct analysis of oil samples after dilution in the gel permeation chromatography eluent without further sample clean-up. Detection limits were about 5 and 10 micrograms/kg with the thermionic and the flame photometric detector, respectively, when using an injection volume of only 30 microliters of the 20-fold diluted oil. The total procedure was linear in the 0.01-10 mg/kg range for both detectors. For twenty organophosphorus pesticides, the relative standard deviations were 3-13% at the 20-60 micrograms/kg level.

On-line high-performance liquid chromatography-solvent evaporation-high-performance liquid chromatography-capillary gas chromatography-flame ionisation detection for the analysis of mineral oil polyaromatic hydrocarbons in fatty foods.

An automated on-line method is described that involves a first LC separation on a large column, evaporation of a 6-ml fraction in an on-line solvent evaporator, a second LC separation using a different mobile phase, fractionating the components of interest and transfer to GC through the in-line vaporizer/overflow interface. The method is designed for the injection of a large amount of food extract (e.g. up to 200 mg of fat) and is applied to the analysis of mineral oil material in a linseed oil.

[Laser-induced thermotherapy of liver metastases with MRI control. Prospective results of an optimized therapy procedure]. / Laserinduzierte Thermotherapie (LITT) von Lebermetastasen unter MRT-Kontrolle. Prospektive Ergebnisse eines optimierten Therapieverfahrens.

In a prospective study MR-guided laser-induced thermotherapy (LITT) of liver metastases was optimized and the clinical parameters evaluated. Seventeen patients (4 women, 13 men) with a total of 29 lesions underwent LITT in 25 sessions through 1 March 1994. Twelve of them suffered from colorectal carcinomas, 5 from other primary tumors. Under local anesthesia a maximum of 5 lesions per patient were treated. For MR-guided LITT a neodymium yttrium aluminum garnet laser (Nd-YAG 1064 nm) with energy varying from 3 to 7.5 W was used. Energy and application time were defined through a computer program; they varied, dependent on the lesion size, location and consistency. To monitor the progress of LITT two special thermosensitive MR sequences (Thermo-Turbo-FLASH and FLASH-2D sequences) were individually optimized, whereby the increase in temperature correlates with an increase in signal loss. All procedures were well tolerated without significant early or late side effects. In only one patient was minimal air documentated in the pleural cavity with spontaneous resorption. Patients were dismissed without complaints after 24-h hospitalization. The control parameters of the contrast-enhanced dynamic MRI documented at optimal response a hypointense signal around the tip of the laser applicator, in accordance with laser-induced necrosis. In lesions 20 mm or smaller, nearly 100% tumor necrosis was achieved and in lesions larger than 20 mm, only 50%. Follow-up evaluation 6 months after treatment shows a significantly better response in lesions with a diameter of 20 mm or smaller, with a local tumor control rate of 66%. In lesions larger than 20 mm only a local tumor control rate of 35% could be achieved. Laser-induced thermotherapy (LITT) represents a new, minimally invasive technique for tumor destruction and a high response rate in small liver metastases.