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PubMed2005

Advanced oxidation protein products (AOPP) and high-sensitive C-reactive protein (hs-CRP) in an "atheroma-free model": Down's syndrome.

Murdoch et al. in 1977 called Down syndrome an "atheroma-free model." In this preliminary study, we investigated advanced oxidation protein products (AOPP) and high-sensitive C-reactive protein (hs-CRP) in 47 age-matched Down syndrome patients and 20 age-matched healthy controls. In these healthy patients, we detected no new biochemical risk factors for atherosclerosis, such as AOPP and hs-CRP, so risks are probably considerably lower.

Adolescent↗
PubMed2006

Purification and characterization of Heteroscorpine-1 (HS-1) toxin from Heterometrus laoticus scorpion venom.

Crude venom from Thai giant scorpion, Heterometrus laoticus, most commonly found in the northeastern area of Thailand, was evaluated for PD50 (paralytic dose 50) activities from abdominal injection to cricket (Gryllus sp.) and activities against various kinds of microorganisms. It exhibited good results in disc diffusion assay for Bacillus subtilis, Klebsiella pneumoniae, Pseudomonas aeruginosa and Staphylococcus aureus. After purification, toxin showed acceptable results for PD50 determination in entrapped cricket as well as inhibitory activity against B. subtilis, K. pneumoniae, and P. aeruginosa with activities over 300 times higher than that of the crude venom. The purified fraction was showed to contain a single band in SDS-PAGE. MALDI-TOF-MS/MS analysis showed one peak of major protein with 8293Da. Partial amino acid sequence show high homology to Scorpine-a polypeptide toxin family with potassium channel blocking and defensin activity. The novel toxin was named "Heteroscorpine-1" (HS-1) as the first Scorpine from genus Heterometrus. After full length determination by PCR, HS-1 gene was found to be composed of two exons, separated by an intron. Deduction revealed 95 amino acid residues with 19 residues as the leading sequence. It showed about 80% similarity to Panscorpine and Opiscorpine.

Amino Acid Sequence↗
PubMed2005

Determination of organochlorine pesticides and their derivations in water after HS-SPME using polymethylphenylvinylsiloxane-coated fiber by GC-ECD.

A porous sol-gel polymethylphenylvinylsiloxane-coated fiber was applied to optimize the headspace solid-phase microextraction (HS-SPME) conditions for the analysis of 11 organochlorine pesticides (OCPs) and their derivations in water. Their determination was carried out using gas chromatography (GC) with electron capture detector (ECD). The method developed was applied to the analysis of lake water samples. Detection limits varied from 0.835 to 13.0 ng/l and relative standard deviations (RSD<or=11.8%) were obtained showing that the precision of the method is reliable. The method also showed good linearity for the tested concentration range with regression coefficients (R2) ranging between 0.980 and 0.999. Recoveries for all the analytes ranged from 71.5% to 115.5%. The results demonstrate the suitability of the HS-SPME/GC-ECD approach for the analysis of multi-residue OCPs in water.

Chromatography, Gas↗
PubMed2002

Improved oral bioavailability of cyclosporin A in male Wistar rats. Comparison of a Solutol HS 15 containing self-dispersing formulation and a microsuspension.

Oral bioavailability of the highly lipophilic and poorly water-soluble immunosuppressive agent cyclosporin A (CyA) in two different formulations was investigated in male Wistar rats. An aqueous microsuspension and a self-dispersing formulation composed of the surface-active ingredients Solutol HS 15:Labrafil M2125CS:oleic acid=7:2:1 (v/v/v) were administered to the animals at a dose level of 20 mg/kg. In order to calculate the absolute oral bioavailability, CyA was additionally administered intravenously at 10 mg/kg as microsuspension. It was found that the oral bioavailability of CyA in the Solutol HS 15-based formulation was twofold higher as compared to the microsuspension (69.9+/-2.8 vs. 35.7+/-3.3%, P=0.001). By contrast, the time to reach maximum plasma concentration (t(max)) and the terminal half-life (t(1/2)) did not differ significantly with the different formulations (t(max): 7.0+/-1.0 vs. 6.3+/-1.7 h; t(1/2): 20.5+/-2.9 vs. 16.7+/-4.7 h). In vitro solubility experiments demonstrated a marked increase in the aqueous solubility of CyA in the presence of the self-dispersing formulation as compared to the micronized powder alone (solubility after 120 min at 37 degrees C: 136 vs. 23.2 microg/ml in human gastric juice; 133 vs. 10.8 microg/ml in simulated intestinal juice). Most likely, the enhanced systemic exposure of CyA in the self-dispersing formulation was caused by improved solubility of CyA in the gastrointestinal fluids in the presence of the surface-active ingredients. Additional factors that may have contributed to increased oral bioavailability are inhibition of metabolism and/or transport processes as well as permeability enhancement by the co-administered excipients.

Administration, Oral↗
PubMed2001

Analysis of fatty acid ethyl esters in hair as possible markers of chronically elevated alcohol consumption by headspace solid-phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS).

Fatty acid ethyl esters (FAEE) are products of the nonoxidative ethanol metabolism, which are known to be detectable in blood only about 24h after the last alcohol intake. After deposition in hair they should be suitable long-term markers of chronically elevated alcohol consumption. Therefore, a method for the analysis of ethyl myristate, ethyl palmitate, ethyl oleate and ethyl stearate from hair was developed based on the extraction of the hair sample by a dimethylsulphoxide (DMSO)/n-hexane mixture, separation and evaporation of the n-hexane phase and application of headspace solid-phase microextraction (HS-SPME) in combination with gas chromatography-mass spectrometry (GC-MS) to the extract. For use as internal standards, the corresponding D(5)-ethyl esters were prepared. The HS-SPME/GC-MS measurements were automatically performed using a multi-purpose sampler. The detection limits of the FAEE were between 0.01 and 0.04ng/mg and the reproducibility was between 3.5 and 16%. By application of the method to hair samples of 21 fatalities with known heavy alcohol abuse 0.045-2.4ng/mg ethyl myristate, 0.35-13.5ng/mg ethyl palmitate, 0.25-7.7ng/mg ethyl oleate and 0.05-3.85ng/mg ethyl stearate were measured. For social drinkers (30-60g ethanol per week), the concentrations were about one order of magnitude smaller. For 10 teetotalers negative results or traces of ethyl palmitate were found. It was shown by supplementary investigations in single cases that FAEE are also present in sebum, that there is no strong difference in their concentrations between pubic, chest and scalp hair, and that they are detectable in hair segments after a 2 months period of abstinence. From the results follows that the measurement of FAEE concentrations in hair is a useful way for a retrospective detection of alcohol abuse.

Adolescent↗
PubMed2002

Home-cage activity in heterogeneous stock (HS) mice as a model of baseline activity.

Behavioral genetic work in humans indicates that clinical hyperactivity is best viewed as the extreme end of activity levels in the population. However, current animal models of hyperactivity are not studied as quantitative traits as they are either knockout models or inbred strains. Furthermore, these animal models generally demonstrate elevated locomotion in novel environments, but not in their home-cages. This is the opposite of the symptoms seen in the human condition where childhood hyperactivity is generally more pronounced in constant, unstimulating situations. In this study we filmed an outbred population of 44 heterogeneous stock (HS) mice under red light during their active phase, to assess the reliability of individual differences in home-cage behavior and extract an index of home-cage activity (HCA) level. We then compared this measure to locomotor behavior in a novel environment--the open-field. Reliable individual differences in home-cage behaviors such as running, swinging on bars, and burrowing were found, and principal component factor analysis yielded a general activity factor, which accounted for 32% of the variance and correlated 0.90 with a subjective impression of activity level. The correlation between HCA and locomotor activity in the open-field was 0.23, which was non-significant. However, the association with HCA level appeared to increase over the five minutes of the open-field, presumably as the mice habituated. Furthermore, although mice displaying particularly high and low HCA were indistinguishable early in the open-field task, they became significantly differentiated over time. We conclude that home-cage behaviors and the open-field, after habituation, display good face and construct validity, and may provide a good model of baseline activity for quantitative trait loci (QTL) discovery and functional genomics in the HS mice.

Animals↗
PubMed1991

Human beta-globin locus control region: analysis of the 5' DNase I hypersensitive site HS 2 in transgenic mice.

The human beta-globin locus control region (LCR) is essential for high-level expression of human epsilon-, gamma-, and beta-globin genes. Developmentally stable DNase I hypersensitive sites (designated HS) mark sequences within this region that are important for LCR activity. A 1.9-kilobase (kb) fragment containing the 5' HS 2 site enhances human beta-globin gene expression 100-fold in transgenic mice and also confers position-independent expression. To further define important sequences within this region, deletion mutations of the 1.9-kb fragment were introduced upstream of the human beta-globin gene, and the constructs were tested for activity in transgenic mice. Although enhancer activity was gradually lost with deletions of both 5' and 3' sequences, a 373-base-pair (bp) fragment retained the ability to confer relative position-independent expression. Three prominent DNase I footprints were observed in this region with extracts from the human erythroleukemia cell line K-562, one of which contained duplicated binding sites for transcription factor AP-1 (activator protein 1). When the 1.9-kb fragment containing an 18-bp deletion of the AP-1 binding sites was tested in transgenic mice, enhancer activity decreased 20-fold but position-independent expression was retained.

Animals↗
PubMed1995

Robotics in the hematology laboratory. An evaluation of the productivity of the Sysmex HS-330.

The productivity of a robotic hematology system (Sysmex HS-330) was compared with that of existing automated, semi-automated, and manual systems (Coulter Counter-Model STKS; Sysmex R-1000 reticulocyte counter. Geometric Data Miniprep slide maker, respectively) in the clinical hematology laboratory of a large hospital. On average, for a batch of 50 samples, the HS-330 performed a blood count with a 5-part differential, a reticulocyte count, and prepare a blood smear 23 minutes faster than could trained technologists using existing equipment. An estimated 1.8 medical technologists could be assigned to other tasks; however, because of the equipment's high cost, in a laboratory processing 1,000 samples per day, 4.6 years would be needed before any actual cost savings were experienced.

Efficiency↗
PubMed2006

HS-SPME determination of volatile carbonyl and carboxylic compounds in different matrices.

Specific chromatographic methodologies are developed for the analysis of carboxylic acids (C(2)-C(6), benzoic) and aldehydes (C(2)-C(10)) of low molecular weight in diverse matrices, such as air, automotive exhaust gases, human breath, and aqueous matrices. For carboxylic acids, the method is based on their reaction with pentafluorobenzyl bromide in aqueous solution, followed by the separation and identification of the resultant pentafluorobenzyl esters by means of headspace (HS)-solid-phase microextraction (SPME) combined with gas chromatography (GC) and electron capture detection (ECD). Detection limits in the microg/m(3) range are reached, with relative standard deviation (RSD) less than 10% and linear response (R(2) > 0.99) over two orders of magnitude. The analytical methodology for aldehydes is based on SPME with simultaneous derivatization of the analytes on the fiber, by reaction with pentafluorophenylhydrazine. The derivatization reagent is previously deposited on the SPME fiber, which is then exposed to the gaseous matrix or the HS of the sample solution. The pentafluorophenyl hydrazones formed on the fiber are analyzed selectively by means of GC-ECD, with detection limits in the ng/m(3) range, RSD less than 10%, and linear response (R(2) > 0.99) over two orders of magnitude.

Aldehydes↗
PubMed1992

Molecular and developmental studies of a sperm acrosome antigen recognized by HS-63 monoclonal antibody.

A conserved mouse sperm antigen (MSA-63) recognized by a monoclonal antibody (HS-63) was isolated from mouse testes by single-step immunoaffinity chromatography. Isolated MSA-63 preparation was shown to be a group of proteins ranging from 24-84 kDa and with isoelectric points (pIs) ranging from 4.0-6.0 when analyzed by two-dimensional (2-D) gel electrophoresis. Microsequencing techniques were employed to determine the relationships of various protein spots on 2-D gels. Partial amino acid sequences of some protein spots in isolated MSA-63 preparation were shown to be homologous to mouse actins, while others revealed homology only to the SP-10 protein. Rabbit antisera raised against isolated MSA-63 antigen preparation were used to immunoscreen a mouse testis cDNA library. Isolated cDNA clones carrying a 1.2-kb insert were used to obtain nucleotide sequences containing open-reading frames and to deduce the corresponding amino acid sequence of MSA-63. A high degree of homology was observed between MSA-63 and a known human sperm antigen, SP-10, at DNA/protein levels. Amino acid sequences of tryptic peptides derived from protein spots of 24-47 kDa and pIs of 4.2-4.4 were found to be identical to those deduced from isolated cDNA clones. The gene expression of MSA-63 during spermatogenesis in mice was studied using a specific cDNA probe as well as HS-63. It was observed that MSA-63 was not expressed until the postmeiotic stages of spermatogenesis.

Acrosome↗
PubMed2001

Hepatocyte-mediated transport to the bile of AMI-HS, a particulate contrast agent.

RATIONALE AND OBJECTIVES: The elimination of hepatocyte-directed particulate contrast agents has not been studied in the same detail as particles eliminated mainly by the mononuclear phagocyte system. The aim of the present study was to elucidate the fate of these particles by a multidisciplinary approach. METHODS: After intravenous injection of AMI-HS particles directed to the hepatocytes, rats were killed and cytological studies, by both electron microscopy and histochemistry, and spectroscopic studies of the bile were performed. The data were compared with a dynamic magnetic resonance study of the heart and liver. RESULTS: The particles were rapidly cleared from the blood by Kupffer cells and hepatocytes and then found first in the vascular and later in the biliary pole of the hepatocytes. After 24 hours, a relaxometric characterization of the bile showed the presence of unchanged particles in the bile. CONCLUSIONS: These results show the capacity of the liver to excrete unchanged AMI-HS particles directly into the bile.

Animals↗
PubMed1989

Sensitivity to ethanol hypnosis and modulation of chloride channels does not cosegregate with pentobarbital sensitivity in HS mice.

Several findings suggest that barbiturates and alcohol produce their sedative effects through a common neural and possibly a common genetic mechanism. We tested this hypothesis by examining the correlation between ethanol and pentobarbital sedative effects in individual animals from a genetically heterogeneous population. The duration of pentobarbital-induced hypnosis (sleep-time) was unrelated to the sleep-time produced by ethanol in heterogeneous stock (HS) mice. Therefore, the present study also examined the effect of ethanol, pentobarbital, and flunitrazepam on muscimol-stimulated chloride flux into brain membranes prepared from HS mice selected for differences in pentobarbital- and ethanol-induced sleep-time. Brain membranes from mice selected for differences in ethanol sleep-time were differentially responsive to ethanol- and flunitrazepam-, but not to pentobarbital-induced augmentation of muscimol-stimulated chloride flux. No differences in augmentation of chloride flux by ethanol, pentobarbital, or flunitrazepam were found in membranes prepared from mice differentially sensitive to pentobarbital hypnosis. The ability of muscimol to stimulate chloride uptake was not related to ethanol or pentobarbital sensitivity. These findings suggest that sensitivity to ethanol is not likely to be genetically linked to pentobarbital sensitivity.

Animals↗
PubMed2005

The HS:1 serostrain of Campylobacter jejuni has a complex teichoic acid-like capsular polysaccharide with nonstoichiometric fructofuranose branches and O-methyl phosphoramidate groups.

Recently, the CPS biosynthetic loci for several strains of Campylobacter jejuni were sequenced and revealed evidence for multiple mechanisms of structural variation. In this study, the CPS structure for the HS:1 serostrain of C. jejuni was determined using mass spectrometry and NMR at 600 MHz equipped with an ultra-sensitive cryogenically cooled probe. Analysis of CPS purified using a mild enzymatic method revealed a teichoic acid-like [-4)-alpha-d-Galp-(1-2)-(R)-Gro-(1-P](n), repeating unit, where Gro is glycerol. Two branches at C-2 and C-3 of galactose were identified as beta-d-fructofuranoses substituted at C-3 with CH(3)OP(O)(NH(2))(OR) groups. Structural heterogeneity was due to nonstoichiometric glycosylation at C-3 of galactose and variable phosphoramidate groups. Identical structural features were found for cell-bound CPS on intact cells using proton homonuclear and (31)P heteronuclear two-dimensional HR-MAS NMR at 500 MHz. In contrast, spectroscopic data acquired for hot water/phenol purified CPS was complicated by the hydrolysis and subsequent loss of labile groups during extraction. Collectively, the results of this study established the importance of using sensitive isolation techniques and HR-MAS NMR to examine CPS structures in vivo when labile groups are present. This study uncovered how incorporation of variable O-methyl phosphoramidate groups on nonstoichiometric fructose branches is used in C. jejuni HS:1 as a strategy to produce a highly complex polysaccharide from its small CPS biosynthetic locus and a limited number of sugars.

Campylobacter jejuni↗
PubMed1985

Comparison and diagnostic value of the coccidioidin heat-stable (HS and tube precipitin) antigens in immunodiffusion.

Coccidioides immitis produces two unrelated heat-stable antigens which are diagnostically useful in immunodiffusion tests. One, the tube precipitin antigen, is valuable for specifically detecting antibody and diagnosing early primary cases of coccidioidomycosis. The other heat-stable antigen, designated HS, is the most useful coccidioidin antigen for specifically immunoidentifying C. immitis cultures. Both of the antigens were compared and evaluated for their usefulness in exoantigen and serologic immunodiffusion tests. Our studies indicated that the detection of tube precipitin antigen is of limited value for immunoidentifying C. immitis isolates because the antigen is common to certain gymnoascaceous saprophytes, such as Arachniotus, Auxarthron, and Malbranchea species, that form alternate arthroconidia. Antibodies to HS antigens are infrequently found in human sera from patients with coccidioidomycosis and are thus of little serodiagnostic value.

Antibodies, Fungal↗
PubMed1978

Comparison of continuous and discontinuous counterimmunoelectrophoresis with immunodiffusion in identification of candida antibody using HS antigen.

We have compared continuous counterimmunoelectrophoresis (CCIE) and discontinuous counterimmunoelectrophoresis (DCIE) with immunodiffusion (ID) in the identification of Candida albicans antibody using HS antigen. Eighteen sera with a titer of 1/1 to 1/32 1/32 by CCIE were titered by CCIE and ID. Fewer were positive by CCIE and ID (13 of 18 and 9 of 18, respectively). Furthermore, the titers were lower when measured by these two methods in comparison to DCIE (4 of 18 and 3 of 18 sera had titers greather than or equal to 1/8 in comparison with 7 of 18 greater than or equal to 1/8 by DCIE). Finally, DCIE was found to be much faster than CCIE and ID. After 20 min of electrophoresis, 17 of 21 sera were positive by DCIE, whereas none were positive by CCIE. Even after 90 min, only 10 of 21 were positive by CCIE, whereas 4 of 21 were positive by ID. The increased sensitivity and speed of DCIE over CCIE and ID suggest that DCIE is preferable to these other methods in the measurement of candida antibody using HS antigen.

Antibodies, Fungal↗
PubMed1998

First experience with the REMEDi HS urine benzodiazepine assay.

Ninety eight urine samples were analysed with an immunoassay benzodiazepine kit. A total of 68 urine specimens that were presumptively positive for benzodiazepines were evaluated by the REMEDi HS urine benzodiazepine assay (BIO-RAD, Munich, Germany). Of this number, 53 (78%) specimens were found by REMEDi to contain one or more benzodiazepines or their metabolites, and 15 (22%) were found to be negative. From the discordant group of 15 samples, eight were found to be negative using conventional chromatographic procedures (HPLC or GC/MS), while seven contained one or more benzodiazepines or metabolites, each of which were below the individual cut-off level specified by the manufacturer. Additionally 30 urine specimens that were negative for benzodiazepines using immunoassay were also tested by REMEDi. Two samples were found to be positive. These results could not be confirmed by other chromatographic techniques. The REMEDi HS benzodiazepine assay can be a very useful complementary technique in the clinical/forensic toxicology laboratory, especially for the identification of the parent benzodiazepines administered. The assay provides a rapid result in emergency situations and is useful in confirmation of preliminary positive immunoassay results.

Automation↗
PubMedDate not supplied

Analysis of the volatile components of five Turkish Rhododendron species by headspace solid-phase microextraction and GC-MS (HS-SPME-GC-MS).

Volatile constituents of various solvent extracts (n-hexane, CH2Cl2, H2O) of 15 different organs (leaves, flowers, fruits) of five Rhododendron species (Ericaceae) growing in Turkey were trapped with headspace solid-phase microextraction (HS-SPME) technique and analyzed by GC-MS. A total of 200 compounds were detected and identified from organic extracts, while the water extracts contained only traces of few volatiles. The CH2Cl2 extract of the R. luteum flowers was found to exhibit the most diverse composition: 34 compounds were identified, with benzyl alcohol (16.6%), limonene (14.6%) and p-cymene (8.4%) being the major compounds. The CH2Cl2-solubles of R. x sochadzeae leaves contained only phenyl ethyl alcohol. This study indicated appreciable intra-specific variations in volatile compositions within the genus. Different anatomical parts also showed altered volatile profiles. This is the first application of HS-SPME-GC-MS on the volatiles of Rhododendron species.

Benzyl Alcohol↗
PubMed1983

Reactivation of VX-inhibited cholinesterase by 2-PAM and HS-6 in rats.

Atropinized rats intoxicated with ethyl-S-2-diisopropyl aminoethyl methyl phosphonothioate (VX), 15 mg/kg iv, were divided into three groups and were treated with normal saline, iv, 30 mg/kg of 2-PAM C1, iv, and 30 mg/kg of HS-6, iv. One hr after administration of therapy they were decapitated and cholinesterase (ChE) activity was determined on blood, brain and diaphragm tissue. Both 2-PAM C1 and HS-6 markedly reactivated VX-inhibited blood and diaphragm ChE. Brain ChE activity was not significantly reactivated by either oxime. The effectiveness of these oximes in restoration of VX-inactivated ChE in vivo offers an explanation as to why conventional atropine/oxime therapy is so effective against VX intoxication.

Animals↗