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PubMed1999

Blind separation of multichannel electrogastrograms using independent component analysis based on a neural network.

The electrogastrogram (EGG) is an abdominal surface measurement of gastric myo-electrical activity which regulates gastric contractions. It is of great clinical importance to record and analyse multichannel EGGs, which provide more information on the propagation and co-ordination of gastric contractions. EGGs are, however, contaminated by myo-electric interference from other organs and artefacts such as motion and respiration. The aim of the study is to separate the gastric signal from noisy multichannel EGGs without any information on the interference, using independent component analysis. A neural-network model is proposed, and corresponding unsupervised learning algorithms are developed to achieve the separation. The performance of the proposed method is investigated using artificial data simulating real EGG signals. Experimental EGG data are obtained from humans and dogs. The processed results of both simulated and real EGG data show the following: first, the proposed method is able to separate normal gastric slow waves from respiratory artefacts and random noises. It is also able to extract gastric slow waves, even when the EGG is contaminated by severe respiratory and ECG artefacts. Secondly, when the stomach contains various gastric electric signals with different frequencies, the proposed method is able to separate these different signals, as illustrated by simulations. These data suggest that the proposed method can be used to separate gastric slow waves, respiratory and motion artefacts, and intestinal myo-electric interference that are mixed in the EGG. It can also be used to detect gastric slow-wave uncoupling, during which the stomach has multiple gastric signals with different frequencies. It is believed that the proposed method may also be applicable to other biomedical signals.

Adult↗
PubMed1996

A rapid method for separation of plasma low and high density lipoproteins for tocopherol and carotenoid analyses.

Ultracentrifugation (UC) is the method most often employed for separation and quantification of lipoproteins. Because this procedure requires expensive laboratory equipment, a large volume of fresh sample and an inordinate amount of time, it may not be ideal for routine clinical/experimental use. The aim of the current study was to evaluate a method which combines selective precipitation (HDL-P) and immunoseparation (LDL-I) for the rapid and reliable isolation of high density lipoproteins (HDL) and low density lipoproteins (LDL) specifically for vitamin E and carotenoid determination within these fractions. Cholesterol and triacylgylcerol concentrations within the HDL and LDL were also determined to enable expression of vitamin E and carotenoid concentrations per gram of lipid. Isolation of lipoproteins by UC was used as the reference method (HDL-UC/LDL-UC). There were no significant differences between methods for alpha- and gamma-tocopherol in LDL and HDL. Carotenoids measured in HDL and LDL were comparable between the methods. The exception was higher lutein/zeaxanthin concentration in HDL-P and LDL-I compared to HDL-UC and LDL-UC, respectively. Additionally, lycopene concentration was significantly lower in LDL-I compared to LDL-UC. In comparing vitamin E and carotenoid values in lipoproteins separated from fresh and frozen plasma by the direct method, there was no difference in alpha-tocopherol or the majority of carotenoids measured. In conclusion, a combination of selective precipitation and immunoseparation of fresh or frozen plasma for subsequent alpha- and gamma-tocopherol analyses provides an accurate and reliable alternative to lipoprotein separation by UC. Additionally, carotenoid concentrations in HDL separated by selective precipitation and analyses of alpha- and beta-carotenes and beta-cryptoxanthin in LDL separated by immunoseparation are also reliable, while lycopene and lutein/zeaxanthin concentrations in LDL-I are not readily comparable to LDL-UC.

Blood Specimen Collection↗
PubMed1987

Rapid separations of diacyl- and dialkylglycerol enantiomers by high performance liquid chromatography on a chiral stationary phase.

Rapid and practical separations of 1,2- and 2,3-diacyl- and dialkyl-sn-glycerol enantiomers as their 3,5-dinitrophenyl-urethane derivatives were carried out by normal-phase high performance liquid chromatography on a chiral stationary phase, N-(R)-1-(alpha-naphthyl)ethylaminocarbonyl-(S)-valine chemically bonded to gamma-aminopropyl silanized silica. Complete separations of the racemates into enantiomers were achieved for both of the diacyl- and dialkylglycerols within 10 min using a stainless steel column (25 cm long) packed with the 5-mu particles, an isocratic elution with a mixture of hexane/ethylene dichloride/ethanol as a mobile phase and a UV detector. The sn-1,2-enantiomers were eluted ahead of the corresponding sn-2,3-enantiomers. Satisfactory separation of the sn-1,3-diacylglycerols from the corresponding enantiomers and the separation of the homologues differing in acyl and alkyl groups were also observed. The formations of hydrogen bonding and charge transfer complex between the urethane derivatives and the stationary phase may contribute to the enantiomer separations.

Acylation↗
PubMed2004

Separation of thoraco-omphalopagus Siamese twin.

A pair of female thoraco-omphalopagus twins, with two pairs of lungs, common diaphragm and separate pleural cavities was separated at the age of 5(1/2) months after a parasitic relationship had developed between them. Before separation both the babies developed recurrent respiratory tract infection and frequent diarrhea. They were treated with medical therapy and made fit for anesthesia. The combined weight of the twins was 4(1/2) kg at birth and the combined weight at the time of separation was 9(1/2) kg. One baby was smaller in growth compared to the other baby. The closure of anterior abdominal wall was difficult in both babies. However, we could close it without use of any biodegradable patches or tissue expander. Both the babies survived and are doing well after 3 months of separation.

Female↗
PubMed2003

Parental separation at birth and criminal behaviour in adulthood--a long-term follow-up of the Finnish Christmas Seal Home children.

BACKGROUND: Parental absence has been connected with later criminality. We studied the association between very early separation and criminality in a unique data set. METHODS: The index cohort consisted of 2,906 subjects born between 1945 and 1965 in Finland who were temporarily isolated from their family immediately after birth and sent to adequate nursing homes due to tuberculosis in the family. The average separation time was 7 months. For every index subject, two reference subjects (matched for sex, year of birth and place of birth) were gathered. Data on criminal offences were obtained from Statistics Finland arising from adolescence to middle age, between January 1, 1977 and December 31, 1998. The association between parental separation and subcategories of non-violent and violent criminality and violent recidivism, respectively, in male and female offspring was analysed. Subjects who had committed at least two violent crimes were defined as violent recidivists. RESULTS: Of the male index subjects, 12.1% as compared with 7.1% of the reference cohort (estimated relative risk RR 1.73; 95% CI 1.42-2.11) had committed violent offences. Of the male index subjects 5.2% and of the male reference subjects 3.6% were violent recidivists (RR 1.47; 1.10-1.98). Of the male index subjects 26.3% and of the reference cohort 23.4% had committed nonviolent crimes (RR 1.14; 1.01-1.29). Among females, non-violent crimes were committed by 7.9% of the index subjects and by 5.0% of the reference subjects, respectively (RR 1.54; 1.18-2.00). Violent crimes were rare among female subjects. CONCLUSION: Criminal behaviour was more prevalent among both male and female subjects separated at birth from their families because of tuberculosis in the family than in the reference cohort. Especially violent crimes were prevalent in the male index cohort. Even so, the differences between the index and reference cohorts were rather modest. Very early separation may have some, although limited, influence on later criminality in the offspring.

Adult↗
PubMed1999

Repeated neonatal maternal separation alters intravenous cocaine self-administration in adult rats.

Behavioural responses to psychostimulant drugs can be profoundly affected by early environmental influences. The aim of this study was to describe the effects of repeated brief separations of rat pups from their dams during the early neonatal period on cocaine self-administration behaviour as adults. Lister hooded rats exposed to a repeated maternal separation procedure (REMS) showed altered acquisition and maintenance of cocaine self-administration as adults, the effects being dose and gender-dependent. Overall, the patterns of acquisition of self-administration across three doses of cocaine (0.05, 0.08 and 0.5 mg/injection) suggested a rightward shift in the acquisition dose-effect functions for the REMS animals relative to control animals. At 0.05 mg/injection, there was a retarded acquisition of cocaine self-administration in male and female neonatally separated rats. At 0.08 mg/injection there was a facilitated acquisition in female neonatally separated subjects. After establishment of stable self-administration of the training dose, in the same cohort of subjects, rightward and downward shifts in the cocaine self-administration dose-effect functions were determined for female and male REMS subjects, respectively, relative to their controls. The dose-effect function for both female groups was shifted to the left of that of the respective male groups, although the lighter body weights of the females meant that they administered a higher unit dose per unit body weight than the males. Whereas male REMS subjects tended to self-administer less cocaine than the controls at the dose eliciting maximal responding (0.03 mg/injection) and to make fewer lever responses overall at each dose tested, female REMS subjects self-administered significantly more cocaine than their respective controls at a dose of 0.03 mg/injection. There was no differential sensitivity to the rate-altering effects of the selective dopamine D2 receptor antagonist, eticlopride, or to the selective dopamine D1 receptor antagonist, SCH 23390. These data provide further evidence that altered early environment affects drug-taking behaviour in a developmentally specific and gender-specific manner, with the effects of neonatal separation contrasting with previously published data on the effects of post-weaning isolation rearing.

Analysis of Variance↗
PubMed2002

Influence of soluble polymer polyvinylpyrrolidone on separation of small peptides and amino acids by microchip-based capillary electrophoresis.

In microchip-based capillary electrophoresis, the resolution and separation efficiency of small peptides and amino acids can be noticeably improved by adding a low molecular weight (30,000) soluble polymer additive, polyvinypyrrolidone in the separation medium. Several separation conditions such as injection time and electrophoretic buffer have been investigated and optimized. Using an electro-stacking scheme, the resolution and separation efficiency of small peptides and amino acids can be enhanced significantly. Under the optimal conditions, the separation of fluorescein isothiocyanate Isomer I-labeled small peptides and amino acids was successfully achieved within 100 s.

Amino Acids↗
PubMed2004

Separation of fatty acid methyl esters by comprehensive two-dimensional supercritical fluid chromatography with packed columns and programming of sampling duration.

Fatty acid methyl esters from various fats and oils were separated by comprehensive two-dimensional supercritical fluid chromatography with conventional packed columns and FID detection. The first dimension was a silica gel column and the second dimension was an ODS column. This combination was largely orthogonal for the separation of fatty acid methyl esters. The first dimension separations were primarily based on the number of double bonds while the second dimension separations were based on the chain length. The highly-ordered chromatograms and improved resolution allowed the easy detection and identification of minor components. Although the first dimension separations were performed under isobaric conditions where the peak width increased in proportion to the retention, the programming of the sampling duration allowed us to maintain the optimum re-injection frequency (3-4 times) per peak into the second dimension and so to minimize the total analysis time without deteriorating the resolution.

Animals↗
PubMed2004

Investigation of the separation of scandium and rare earth elements from red mud by use of reversed-phase HPLC.

A chromatographic method has been developed for separation and determination of scandium (Sc) and rare earth elements (REEs) in samples from a red mud (RM)-utilization process. Reversed-phase high-performance liquid chromatography (RP-HPLC) with post-column derivatization using 4-(2-pyridylazo)-resorcinol (PAR) and UV-visible detection at 520 nm was tested using different gradient elution profiles and pH values to optimize separation and recovery, primarily for Sc but also for yttrium and the individual lanthanides, from iron present in the samples. The separation was performed in less than 20 min by use of a mobile phase gradient. The concentration of alpha-hydroxyisobutyric acid ( alpha-HIBA), as eluent, was altered from 0.06 to 0.4 mol L(-1) (pH 3.7) and 0.01 mol L(-1) sodium salt n-octane sulfonic acid (OS) was used as modifier. Very low detection limits in the nanogram range and a good resolution for Sc and REEs except for Y/Dy were achieved. Before application of the method to the red mud samples and to the corresponding bauxites, Sc and REEs were leached from red mud with 0.6 mol L(-1) HNO(3) and mostly separated, as a group, from the main elements by ion exchange/selective elution (6 mol L(-1) HNO(3)) in accordance with a pilot-plant process developed in this laboratory. After evaporation of the eluent to dryness the extracted elements were re-dissolved in the mobile phase. By use of this chromatographic method Sc, which is the most expensive of the elements investigated and occurs in economically interesting concentrations in red mud, could be separated not only from co-existing Fe but also from Y/Dy, Yb, Er, Ho, Gd, Eu, Sm, Nd, Pr, Ce and La. All the elements investigated were individually recovered. Their recoveries were found to be nearly quantitative.

Journal Article↗
PubMed2005

Microemulsion liquid chromatographic method for characterisation of fosinopril sodium and fosinoprilat separation with chemometrical support.

The properties of the eluent are the essential factors governing the efficiency in the high-performance liquid chromatography (HPLC) method. A novel approach in retention modelling in the liquid chromatographic separation of fosinopril sodium and its degradation product, fosinoprilat, applying a microemulsion as the mobile phase, was used. The modifications of the mobile phase included the changes to the type of the lipophilic phase, the type and concentration of co-surfactant and surfactant, as well as the pH of the mobile phase. In this study, a full factorial 2(3) design, as the optimal method for screening of the experiment, was applied for selecting factors which had an influence on separation. Optimisation was done by a central composite design. An appropriate resolution with reasonable retention times was obtained with a microemulsion containing 0.9% w/w of cyclohexane, 2.2% w/w of sodium dodecyl sulphate (SDS), 8.0% w/w of n-butanol and 88.9% of aqueous 25 mM disodium phosphate, the pH of which was adjusted to 2.8 with 85% orthophosphoric acid. Separations were performed on an X-Terra 50-mmx4.6-mm, 3.5-mum particle size column at 30 degrees C. UV detection was performed at 220 nm and with a flow rate of 0.3 mL min(-1). The established method was validated and applied for analysis of appropriate tablets. The proposed chromatographic procedure for the separation of fosinopril sodium and its degradation product is less expensive compared with the conventional reversed-phase HPLC method, as well as being simple and rapid. The optimised and validated method can be used for separation, identification and simultaneous determination of fosinopril sodium and fosinoprilat in bulk drug and in pharmaceutical dose forms.

Chromatography, High Pressure Liquid↗
PubMed2005

Performance and selectivity of polymeric pseudostationary phases for the electrokinetic separation of amino acid derivatives and peptides.

Two polymeric pseudostationary phases, one an acrylamide polymer and the second a siloxane polymer, have been investigated for the separation of naphthalene-2,3-dicarboxaldehyde (NDA)-derivatized amino acids and small peptides. The dervatized amino acids were detected by UV absorbance and laser-induced fluorescence (LIF) detection. The polymers provided very high efficiency and good selectivity for the separation of the amino acids. The separation selectivity using the polymers was significantly different from that of SDS micelles, and there were subtle differences in selectivities between the polymers. Although very good detection limits were obtained with LIF detection, a significant background signal was observed when the polymers were not washed to remove fluorescent impurities. The polymers did not separate the peptides very well. It is postulated that the fixed covalent structure of the polymers prevents them from interacting strongly or efficiently with the peptides, which are large in relation to the analytes typically separated by electrokinetic chromatography using polymers.

Acrylamides↗
PubMed2005

Separation of linear gramicidins using carbon dioxide-containing mobile phases.

Packed-column supercritical-fluid chromatography (pSFC) is presented as a novel method for separating and analyzing gramicidin samples. By use of methanol-modified carbon dioxide as a mobile phase the pentadecapeptides gramicidin A (gA), gramicidin B (gB), and gramicidin C (gC) are readily separated and eluted from a PRP-1 poly(styrene-divinylbenzene) column. Although optimum separation conditions are typically achieved near a column temperature of 40 degrees C, a column pressure of 11 MPa, and 30% methanol modifier, pressure and modifier gradients around these values are also found to improve the overall separation time. Measurements indicate that the mobile phase solubility of gramicidin under these conditions is 5.0+/-0.4 microg mL(-1). Collection of individual peaks during chromatography achieved analytical-scale isolation of 2 microg refined gC from 20 microg injected gramicidin D. Further, supercritical-fluid extraction of 200 microg gramicidin D from a Chromosorb 102 support packed into the vessel produced 57 microg gA in 90% purity. The results establish that carbon dioxide-based mobile phases can be successfully used for the separation of individual gramicidin species.

Carbon Dioxide↗
PubMed2006

Isocratic separation of ginsenosides by high-performance liquid chromatography on a diol column at subambient temperatures.

An improved high-performance liquid chromatographic method for separation of a number of ginsenosides has been developed. The influence of temperature (from 0 to 25 degrees C) on the retention and separation of the ginsenosides was studied by applying a binary mobile phase (acetonitrile/water, 82:18 v/v) and a diol column (LiChrospher 100 Diol). The column temperature is one of the more important parameters for the retention and separation of the components investigated. Selected thermodynamic parameters, including changes of enthalpy (deltaH degrees) and entropy (deltaS degrees), were estimated from linear van't Hoff plots, and possible retention mechanisms were discussed. Moreover, the best separation conditions were selected based on optimization criteria including maximum retention time (t(R max)), minimum resolution (R(s min)), and relative resolution product (r). Temperature regions close to 14 degrees C offered the highest selectivity and almost equal distribution of the ginsenosides peaks across the chromatogram. Under such isocratic conditions, excellent separation of chromatographic standards and selected ginseng samples was achieved in less than 16 min.

Chromatography, High Pressure Liquid↗
PubMed2006

Development of an on-line immobilized-enzyme reversed-phase HPLC method for protein digestion and peptide separation.

This paper describes use of a novel glass bead-based immobilized-enzyme micro column for simple and swift on-line protein digestion then peptide separation by reversed-phase HPLC. The inexpensive and easily made immobilized-enzyme micro column was prepared from aminopropyl controlled-pore glass that was reacted first with glutaraldehyde then with trypsin in the presence of phosphate buffer. Tryptic digestion of bovine serum albumin (BSA) was achieved simply by passing pretreated protein solution through the laboratory made immobilized-trypsin column; the tryptic fragments were then separated by reversed-phase HPLC. The peptide separation was found to be identical to separation of a sample which had undergone conventional enzymatic protein digestion in solution. Digestion of BSA by the immobilized-trypsin column decreased with increasing flow rate of the solution through the column, and 1.0 muL min(-1) was found to be the optimum flow rate for on-line protein digestion with our system. It was also found that the sample required pretreatment with urea before injection, because of a change in the properties of the protein in the presence of urea, and the immobilized-trypsin column lost its function in the presence of acetonitrile. This on-line proteomics system enables simple and rapid protein digestion and was successfully applied to partially micro two-dimensional (2D) chromatographic separation of proteins.

Chromatography, High Pressure Liquid↗
PubMed2006

Pretreatment and one-shot separating analysis of whole catecholamine metabolites in plasma by using LC/MS.

Catecholamines are biogenic amines that play an important role in the nervous system. Some catecholamines have been used as tumor makers of phenochromocytoma, paraganglioma and neuroblastoma. The analysis of total catecholamine metabolites should be useful for one-shot screening of multiple aspects of diseases; however, it is difficult to do this, because the catecholamine metabolites are divided into three groups: five amines, one amino acid and three carbonic acids. Catecholamines and small molecules were separated from plasma proteins by an internal-surface reversed-phase column (protein-coated octadeyclsilica column) and were analyzed by liquid chromatography (LC)/mass spectrometry (MS) using electrospray ionization time-of-flight MS. Using a reversed-phase column and hydrophilic mobile phases, we succeeded in the separation of nine catecholamines, all of which had similar structures. These nine substances were eluted in the following order: norepinephrine, epinephrine, normetanephrine, dopamine, metanephrine, 3,4-dihydroxyphenylalanine, vanillomandelic acid, 3,4-dihydroxyphenylacetic acid and homovanillic acid. The reproducibility of this method was acceptable. The highest coefficient of variation was 7.4%. In addition, various types of compounds were separated from and detected in plasma proteins by applying LC/MS. The plasma direct injection method, which uses an internal-surface reversed-phase column and an ion-pair reagent, allowed us to separate small molecules from plasma proteins. MS detected some compounds that high-performance LC could not succeed in separating and detecting with UV detection. We think that the method can be applied to find new markers in neuroblastoma, by comparing the plasma of patients with that of normal infants. The method can be also used to help in making a diagnosis of other diseases and finding their new makers.

Catecholamines↗
PubMed2006

Magnetic particles for the separation and purification of nucleic acids.

Nucleic acid separation is an increasingly important tool for molecular biology. Before modern technologies could be used, nucleic acid separation had been a time- and work-consuming process based on several extraction and centrifugation steps, often limited by small yields and low purities of the separation products, and not suited for automation and up-scaling. During the last few years, specifically functionalised magnetic particles were developed. Together with an appropriate buffer system, they allow for the quick and efficient purification directly after their extraction from crude cell extracts. Centrifugation steps were avoided. In addition, the new approach provided for an easy automation of the entire process and the isolation of nucleic acids from larger sample volumes. This review describes traditional methods and methods based on magnetic particles for nucleic acid purification. The synthesis of a variety of magnetic particles is presented in more detail. Various suppliers of magnetic particles for nucleic acid separation as well as suppliers offering particle-based kits for a variety of different sample materials are listed. Furthermore, commercially available manual magnetic separators and automated systems for magnetic particle handling and liquid handling are mentioned.

Automation↗
PubMed2004

A new model for separation between brain dopamine and serotonin transporters in 123I-beta-CIT SPECT measurements: normal values and sex and age dependence.

123I-beta-CIT is a radioactive ligand for single-photon emission computed tomography (SPECT) imaging of the pre-synaptic (transporter) re-uptake sites for dopamine (DAT) and serotonin (5HTT), and it is widely used to visualize monoamine turnover. Since 123I-beta-CIT uptake occurs at 5HTT and DAT sites in conjunction with the presence of freely soluble 123I-beta-CIT in brain tissue, adequate separation of these three components is necessary. However, only partial separation is possible with current methods. Two main strategies have previously been used for 123I-beta-CIT component separation, based on the following considerations: (1) the faster uptake rate for 5HTT compared with DAT enables temporal separation by performing 5HTT imaging at 1-2 h and DAT imaging at 20-24 h; (2) blocking the 5HTT re-uptake with citalopram renders 123I-beta-CIT imaging DAT (non-5HTT) specific. In a new analytical model, we combined these two approaches with methods to isolate the passively dissolved 123I-beta-CIT in brain tissue from the monoamine transporter uptake, and to correct the 5HTT and DAT values for concomitant uptake. The new analytical model was used to study brain 5HTT and DAT in 23 normal subjects, with the aim of clarifying the effect of age and sex. A significant correlation between 5HTT and DAT values was found only in the thalamus, indicating successful component separation. Negative correlations between age and DAT were found for basal ganglia, thalami, brain stem and temporal lobes, but not for the frontal, parietal or occipital regions. No correlation with age was found for 5HTT. We found no sex difference for 5HTT or DAT.

Adult↗
PubMed2006

Value of PET/CT versus PET and CT performed as separate investigations in patients with Hodgkin's disease and non-Hodgkin's lymphoma.

PURPOSE: The aim of this study was to assess the clinical benefit of combined [(18)F]FDG PET/CT in patients with malignant lymphoma as compared to separately performed PET and CT. METHODS: Overall, 100 patients with Hodgkin's disease (HD) or non-Hodgkin's lymphoma (NHL) were included in this study. Co-registered PET/CT with [(18)F]FDG and contrast medium was performed in 50 consecutive patients with NHL (n=38) or HD (n=12) for initial staging (IS) (n=12) or re-treatment staging (RS) (n=38). Another 50 patients with NHL (n=32) or HD (n=18) underwent separate PET and CT investigations within a time frame of 10 days for IS (n=22) or RS (n=28). Lymphoma involvement was separately evaluated for seven different regions in each patient. Each patient had clinical follow-up evaluation for >6 months. PET and CT data were analysed separately as well as side-by-side or in fused mode. RESULTS: In the PET/CT group, region-based evaluation for lymphoma involvement suggested a sensitivity/specificity of 85%/91% for CT, 98%/99% for PET and 98%/99% for PET/CT. In the PET and CT group, region-based evaluation showed a sensitivity/specificity of 87%/80% for CT, 98%/99% for PET and 98%/100% for PET and CT read side by side. CONCLUSION: PET was superior to CT alone and was improved further by side-by-side reading of both examinations. However, no significant difference was observed between PET/CT and separate PET and CT imaging in patients with lymphoma.

Adult↗