Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.10.2 (focal adhesion kinase)
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A method for the selective quantitative determination of inorganic arsenic [As(III) + As(V)] in seafood was developed. In order to do so, various procedures for the solubilization and extraction of inorganic arsenic quoted in the literature were tested. None provided satisfactory recoveries for As(III) and As(V) in real samples. Consequently, a methodology was developed which included solubilization with HCl and subsequent extraction with chloroform. The arsenic was solubilized in 9 mol l-1 hydrochloric acid. After reduction by hydrobromic acid and hydrazine sulfate, the inorganic arsenic was extracted into chloroform, back-extracted into 1 mol l-1 HCl, dry-ashed, and quantified by hydride generation-atomic absorption spectrometry (HG-AAS). The analytical features of the method are as follows: detection limit, 3.07 ng g-1 As (fresh mass); precision (RSD), 4.0%; recovery, As(III) 99%, As(V) 96%. In the optimized conditions, other arsenic species--dimethylarsinic acid (DMA), arsenobetaine (AB), arsenocholine (AC) and tetramethylarsonium-ion (TMA+)--were not co-extracted. However, different percentages of minor species were extracted with chloroform: monomethylarsonic acid (MMA) 100%, and trimethylarsine oxide (TMAO) 3-10%. Real samples and reference materials of seafood (DORM-1, DORM-2, TORT-2, CRM-278 and SRM-1566a) were analyzed. The analysis of DORM-1 provided an inorganic arsenic value of 124 +/- 4 ng g-1 As, dry mass (dm), which is very close to the value obtained by other authors using high performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS) and ionic chromatography-hydride generation-atomic absorption spectrometry (IC-HG-AAS).
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PMID:Optimization of the solubilization, extraction and determination of inorganic arsenic [As(III) + (As(V)] in seafood products by acid digestion, solvent extraction and hydride generation atomic absorption spectrometry. 1060 84

An optimised BCR three steps sequential extraction procedure (BCR SEP) and several single extractions with KCl, NH(4)Cl, Na(4)P(2)O(7) and 0.5 mol L(-1) HCl were used for the fractionation of Al, Cu, Fe, Mn, Pb and Zn in CRMs and in samples from a mining area with sulphidic deposits. A good interlaboratory comparability was obtained for Cu, Pb and Zn in CRM 483, CRM 701, SRM 2710 and SRM 2711 by BCR SEP. The reliability of the results obtained is also very satisfactory. Some differences were found between our results and the indicative data for Al and Fe fractionation. However, serious discrepancies were found for Mn, not only for individual steps of the fractionation, but for the data obtained overall (sum of 1-3 steps), and for the total concentration as well. Our results could be utilized as a contribution to the existing indicative values for CRM 483, SRM 2710 and SRM 2711 for interlaboratory study. Moreover, data for the fractionation of elements mentioned above for CRM 701 are first presented here.A high correlation between 0.5 mol L(-1) of HCl-extractable amounts of the elements studied, and the sum of the three steps of BCR SEP in acid sulphatic weathering products and naturally acidified soils was established, which allows us to suggest this rapid and cost-effective single extraction procedure as a valuable tool in contamination assessment.
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PMID:Fractionation of various elements in CRMs and in polluted soils. 1498 15

The aim of the present study was to evaluate in vitro data on platelets collected by apheresis, processed on a preparation set followed by photochemical treatment (PCT). Fifteen single-donor platelet concentrates (PCs) were collected by apheresis (COM.TEC blood cell separator, Fresenius, Bad Homburg, Germany). The platelets were transferred to the preparation set and plasma was removed after centrifugation to resuspend the platelets in approximately 37% plasma and 63% platelet additive solution InterSol. PCT was done by exposing the platelets to amotosalen HCl followed by illumination with ultraviolet light. Blood cell counts and in vitro PLT function were measured up to 5 days. An average of 3.44 +/- 0.28 x 10(11) platelets were collected in a product volume of 351 +/- 21 mL. Plasma removal resulted in a mean platelet loss of 7.8%. After PCT, a progressive decrease in platelet function was observed. LDH level rose through storage (171 +/- 81 U/L) to levels approximating LDH levels observed post-collection (180 +/- 103 U/L). There was a gradual decrease of the platelets to respond to hypotonic shock response from 90 +/- 9 % post-plasma reduction to 48 +/- 16% at day 5. All PLT units met the European requirements for leukoreduction and the pH limit of 6.8 up to day 5 post-collection. The new preparation set was capable of producing platelet units meeting the requirements for PCT. Despite differences observed in in vitro platelet function parameters, PLTs at storage day 5 fit the German and European guidelines.
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PMID:In vitro evaluation of COM.TEC apheresis platelet concentrates using a preparation set and pathogen inactivation over a storage period of five days. 1559 47

A continuous assay using internally quenched fluorescent peptides with the general sequence Abz-peptidyl-(Dnp)P-OH (Abz = ortho-aminobenzoic acid; Dnp = 2,4-dinitrophenyl) was optimized for the measurement of angiotensin I-converting enzyme (ACE) in human plasma and rat tissues. Abz-FRK(Dnp)P-OH, which was cleaved at the Arg-Lys bond by ACE, was used for the enzyme evaluation in human plasma. Enzymatic activity was monitored by continuous recording of the fluorescence (lambda ex = 320 nm and lambda em = 420 nm) at 37 degrees C, in 0.1 M Tris-HCl buffer, pH 7.0, with 50 mM NaCl and 10 microM ZnCl2. The assays can be performed directly in the cuvette of the fluorimeter and the hydrolysis followed for 5 to 10 min. ACE measurements in the plasma of 80 healthy patients with Hip-His-Leu and with Abz-FRK(Dnp)P-OH correlated closely (r = 0.90, P < 0.001). The specificity of the assay was demonstrated by the complete inhibition of hydrolysis by 0.5 microM lisinopril or captopril. Abz-FRK(Dnp)P-OH cleavage by ACE was monitored in rat lung, kidney, heart, and liver homogenates in the presence of a cocktail of inhibitors containing trans-epoxy-succinyl-L-leucylamido-(4-guanido)-butene, pepstatin, phenyl-methylsulfonyl fluoride, N-tosyl-L-phenylalanyl-chloromethyl ketone, and N-tosyl-lysyl-chloromethyl ketone to prevent undesirable hydrolysis. ACE activity in lung, heart and kidney homogenates, but not in liver homogenates, was completely abolished by 0.5 microM lisinopril or captopril. The advantages of the method are the procedural simplicity and the high sensitivity providing a rapid assay for ACE determinations.
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PMID:A continuous fluorescent assay for the determination of plasma and tissue angiotensin I-converting enzyme activity. 1593 79

This paper describes a novel hydride atomizer based on atmospheric pressure dielectric barrier discharge (DBD) plasma. The plasma was generated with a 3700-V, 20.3-kHz, and 5-W electrical power supply and easily sustained with inert gases (He or Ar) at a flow rate of 250 mL.min(-1) after optimization. However, it cannot be sustained with N2. This atomizer offers the advantages of low operation temperature and low power consumption in comparison with the currently used electrothermal quartz atomization operated at 900 degrees C with a power supply of several hundred watts. To confirm the utility of the proposed atomizer, four arsenic species (As(III), As(V), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA)) were determined by the present atomization technique. A hyphenation of HPLC coupled to hydride generation AAS with the optimized DBD atomizer has been successfully used for the speciation of arsenic in order to demonstrate the potential of this atomizer in the present study. The characteristics of the DBD atomizer and the effects of different parameters (discharge gas, gas flow rate, voltage, HCl concentration, KBH4 concentration) are discussed in the paper. Compared with other hydride atomization techniques, the proposed method shows the following features: (1) small size (70 mm x 15 mm x 5 mm), which is preferable for the miniaturization of the total analytical system; (2) low power consumption (< or =5 W), which indicates the possibility of the development of portable, fieldable analytical instrumentation for in situ detection using battery as power supply; (3) low atomizer temperature (approximately 70 degrees C), which is in favor of the compactness of the total instruments; (4) avoidance of residue moisture removal in comparison with the existed GD system, which leads to the facility of the system. The analytical figures of the present technique were evaluated. The detection limits of As(III), As(V), MMA, and DMA obtained with HG-DBD-AAS were 1.0, 11.8, 2.0, and 18.0 microg.L(-1), respectively. The accuracy of the system was verified by the determination of arsenic in reference material of orchard leaves SRM 1571. The concentration of As determined by the present method agreed well with the reference values. The speciation of arsenic in the freeze-dried urine SRM 2670 were carried out, and the results obtained were in agreement with the results of HPLC-ICPMS and the reported values by other laboratories.
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PMID:Atomization of hydride with a low-temperature, atmospheric pressure dielectric barrier discharge and its application to arsenic speciation with atomic absorption spectrometry. 1644 62

16 alpha-[(18)F]fluoro-17beta-estradiol ([(18)F]FES) is a radiotracer for imaging estrogen receptors by positron emission tomography. We developed a clinically applicable automatic preparation system for [(18)F]FES by modifying a cassette-type [(18)F]fluorodeoxyglucose synthesizer. Two milligrams of 3-O-methoxymethyl-16,17-O-sulfuryl-16-epiestriol in acetonitrile was heated at 105 degrees C for 10 min with dried [(18)F]fluoride. The resultant solution was evaporated and hydrolyzed with 0.2 N HCl in 90% acetonitrile/water at 95 degrees C for 10 min under pressurized condition. The neutralization was carried out with 2.8% NaHCO(3), and then the high-performance liquid chromatography (HPLC) purification was performed. The desired radioactive fraction was collected and the solvent was replaced by 10 ml of saline, and then passed through a 0.22-microm filter into a pyrogen-free vial as the final product. The HPLC purification data demonstrated that [(18)F]FES was synthesized with a yield of 76.4+/-1.9% (n=5). The yield as the final product for clinical use was 42.4+/-3.2% (n=5, decay corrected). The total preparation time was 88.2+/-6.4 min, including the HPLC purification and the solvent replacement process. The radiochemical purity of the final product was >99%, and the specific activity was more than 111 GBq/micromol. The final product was stable for more than 6 h in saline containing sodium ascorbate. This new preparation system enables us to produce [(18)F]FES safe for clinical use with high and reproducible yield.
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PMID:Automatic synthesis of 16 alpha-[(18)F]fluoro-17beta-estradiol using a cassette-type [(18)F]fluorodeoxyglucose synthesizer. 1654 84

We have developed a fully automatic method for the synthesis of 16alpha-[18F]fluoroestradiol ([18F]FES) using a disposable cassette system and conventional [18F]FDG module. [18F]FES was synthesized using a GE TracerLab MX module and a modified module control program. Following [18F]fluorination, we hydrolyzed the product three times with a mixture of 2N HCl and CH(3)CN. After HPLC purification, the decay corrected radiochemical yield of [18F]FES was 45.3+/-2.8%, which was stable to 98.2+/-0.2% at 6h after synthesis. This new automated synthesis method provides high and reproducible yields with the advantage of a disposable cassette system.
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PMID:The automatic production of 16alpha-[18F]fluoroestradiol using a conventional [18F]FDG module with a disposable cassette system. 1696 65

The biosorption of copper(II), lead(II), iron(III) and cobalt(II) on Bacillus sphaericus-loaded Diaion SP-850 resin for preconcentration-separation of them have been investigated. The sorbed analytes on biosorbent were eluted by using 1 mol L(-1) HCl and analytes were determined by flame atomic absorption spectrometry. The influences of analytical parameters including amounts of pH, B. sphaericus, sample volume etc. on the quantitative recoveries of analytes were investigated. The effects of alkaline, earth alkaline ions and some metal ions on the retentions of the analytes on the biosorbent were also examined. Separation and preconcentration of Cu, Pb, Fe and Co ions from real samples was achieved quantitatively. The detection limits by 3 sigma for analyte ions were in the range of 0.20-0.75 microg L(-1) for aqueous samples and in the range of 2.5-9.4 ng g(-1) for solid samples. The validation of the procedure was performed by the analysis of the certified standard reference materials (NRCC-SLRS 4 Riverine Water, SRM 2711 Montana soil and GBW 07605 Tea). The presented method was applied to the determination of analyte ions in green tea, black tea, cultivated mushroom, boiled wheat, rice and soil samples with successfully results.
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PMID:Biosorption of copper(II), lead(II), iron(III) and cobalt(II) on Bacillus sphaericus-loaded Diaion SP-850 resin. 1738 50

Atypical antipsychotics such as olanzapine have high affinity for multiple monoamine neurotransmitter receptors and are the mainstay of pharmacological therapy for treatment of schizophrenia. In addition to blocking monoamine receptors, these drugs also affect intracellular signaling cascades. We now report that 24-h treatment with 300 nM olanzapine causes desensitization of serotonin (5-HT)(2A) receptors in A1A1v cells, a rat cortical cell line, as indicated by a reduction in inositol phosphate accumulation following stimulation with a 5-HT(2A/2C) receptor agonist (-)-1-(2,5-dimethoxy-4-lodophenyl)-2-aminopropane HCl. Olanzapine treatment for 24 h increased the levels of 5-HT(2A) receptors in both cytosol (234 +/- 34% of control level) and membrane fractions (206 +/- 14% of control levels) and RGS7 proteins in both cytosol (193 +/- 32% of control levels) and membrane fractions (160 +/- 18% of control levels) as measured on Western blots. Increased phosphorylation of Janus tyrosine kinase (JAK) 2 and increased phosphorylation and nuclear translocation of signal transducer and activator of transcription (STAT) 3 with 24-h olanzapine treatment demonstrate activation of the JAK-STAT signaling cascade. Pretreatment with a JAK inhibitor, AG490 [alpha-cyano-(3,4-dihydroxy)-N-benzylcinnamide], prevented the olanzapine-induced increase in membrane RGS7 protein levels; AG490 alone had no effect on RGS7 protein levels. We verified that treatment with AG490 reduced phosphorylation of JAK2 and inhibited the nuclear localization of phospho-STAT3. Interestingly, treatment with the JAK inhibitor had no effect on 5-HT(2A) receptor protein levels. These data suggest that olanzapine-induced activation of the JAK-STAT signaling cascade causes increased expression of RGS7 protein, which in turn could mediate desensitization of 5-HT(2A) receptor signaling caused by olanzapine because RGS7 binds to Galpha(q) protein and accelerates GTP hydrolysis.
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PMID:Olanzapine increases RGS7 protein expression via stimulation of the Janus tyrosine kinase-signal transducer and activator of transcription signaling cascade. 1739 3

As the film formation of the dry coating process differs completely from conventional coating methods it is of certain interest to define a parameter like the minimum film formation temperature (MFT) used for aqueous dispersion based processes in order to describe an efficient film formation. Film formation occurs mainly during the curing step following the coating phase. Therefore, the film formation of the dry coating process was analyzed with regard to the two process parameters influencing film formation, namely curing temperature and time. Theophylline pellets were coated using the enteric polymer HPMCAS and TEC/Myvacet as plasticizer composition. The polymer and the plasticizer were applied to the pellets simultaneously, except for the beginning of the coating step when the plasticizer has been sprayed 30 s before powder feeding was started. The coated pellets were cured for five different time periods at eight different temperatures. Drug release was determined in 0.1 N HCl. Their surface and cross-sectional morphologies were examined by scanning electron microscopy. The glass transition temperature of the obtained films as well as of the polymer plasticizer mixture obtained by casting a film using an organic solution of the coating components was determined by thermomechanical analysis. At higher curing temperature and/or extended curing time an enhancement of acid resistance is observed. The glass transition temperature of the coating mixture was determined to be 51.7+/-3.3 degrees C. It is close to the temperature needed for film formation. Enteric resistant pellets are obtained after curing for 0.75 h at 55 degrees C. However, enteric resistance was achieved as well slightly below the glass transition temperature at 45 degrees C applying long curing periods of at least 12 h. This may be caused by the plasticizer gradient along the coat which is caused by spraying the plasticizer 30 s before starting powder feeding. This results in a higher plasticizer concentration of the inner layers of the coat relatively to the outer ones. Due to this film formation starts at the inner layers even below the glass transition temperature. As the plasticizer diffuses to the outer layers during curing according to the plasticizer gradient, film formation proceeds.
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PMID:Characterization of the film formation of the dry coating process. 1745 28


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