Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.4.1.14 (SPS)
 813 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A sensitive and selective bioanalytical method for diclofenac using reversed-phase HPLC and fluorescence detection is described. Diclofenac was detected as its fluorescent derivative after on-line post-column photoderivatization. Irradiation with UV light of diclofenac in aqueous solutions leads to the sequential loss of both chlorine substituents and ring closure. The major product, carbazole-1-acetic acid, was detected by a fluorescence detector using an excitation wavelength of 286 nm and an emission wavelength of 360 nm. The self-made reactor was a crocheted ethylene and tetrafluoroethylene (ETFE, named TEFZEL) capillary, 20 m in length, wound directly around a 253.7 nm UV lamp. The capillary was crocheted in order to overcome peak widening. Chromatographic separation was achieved by using a Regis SPS 100 RP-8 column (5 microm; 150 mm x 4.6 mm i.d.) and a LiChrospher 100 RP-18 (5 microm) guard column from E. Merck. The detection limit was 1 ng ml(-1) at an injection volume of 20 microl. Daily relative standard deviations (RSD) were 5.5%, (73 ng diclofenac/ml, n = 9), and 5.1% (405 ng diclofenac/ml, n = 6), respectively. Chromatograms of human aqueous humor and human serum containing diclofenac, and figures showing the time dependent increase/decrease of the photoderivatization product, are shown.
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PMID:Crocheted ETFE-reactor for on-line post-column photoderivatization of diclofenac in high-performance liquid chromatography. 950 51

The chromatographic behavior of an alkyl-diol silica (ADS, 25 x 4 mm I.D.) and a semipermeable surface (SPS, 10 x 10 mm I.D.) supports two types of restricted-access media (RAM), which served as precolumns in column-switching systems for direct injection of large volumes of plasma samples (500 microl), was studied with regard to peak performance, retention and column back pressure. The adsorption of matrix proteins both on sealings (porous frits and sieves) and packings was also examined. Columns of ADS and SPS were unchanged after the injection of 10-20 ml human plasma under normal working conditions. Even when changes occurred on the precolumns (>50 ml of plasma in total), it was still possible to regenerate the column performance by replacing the column sieves, or by washing and removing columns from the system for a period, since the changes were more related to the blockage of sealings and/or the adsorption of proteins on the hydrophilic surfaces. Proteins could eventually be unspecifically adsorbed on the hydrophobic ligand of the support. It was found on one ADS column that the retention decreased by 20% and the pressure increased 30 bar after an intensive loading of 75 ml plasma (injection volume, 500 microl) without reconditioning procedure. Studies showed that the column sealings played the most important role for the lifetime of RAM columns. For ADS columns, using sieves without polytetrafluoroethylene (PTFE) nets were the best. No significant difference in column life span between SPS and ADS was found.
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PMID:Evaluation of liquid chromatographic behavior of restricted-access media precolumns in the course of direct injection of large volumes of plasma samples in column-switching systems. 951 77

Early during fruit ripening in kiwifruit (Actinidia deliciosa var. deliciosa [A. Chev.], C.F. Liang and A.R. Ferguson cv. Hayward), starch is broken down to sucrose and hexose sugars. Concomitantly, sucrose-phosphate synthase (SPS, EC 2.3.1.14) activity measured with saturating substrate increased, suggesting that SPS is induced in response to a higher requirement for sucrose synthesis. A 2584 bp long partial cDNA clone encoding SPS was isolated from ripening kiwifruit. cDNA fragments encoding the 5' end were isolated by PCR, and sequencing revealed at least four closely related (> 96% identity) mRNAs expressed early in kiwifruit ripening. Southern hybridisations in a diploid relative of kiwifruit, Actinidia chinensis (Planch.) var. chinensis, were consistent with the presence of a small gene family. Western analysis indicated a 125 kDa SPS protein present in all tissues of A. chitensis at all stages of development. Steady-state levels of SPS mRNA in A. chinensis increased near fruit maturity as net starch degradation began on the vine, and increased again during ethylene treatment of fruit after harvest. After removal from ethylene SPS transcript levels decreased, only to increase again as fruit moved into the climacteric and starch breakdown was completed. Exposure to low temperatures also caused an increase in SPS transcript level. These results indicate that SPS mRNA increases in kiwifruit in response to the presence of new substrate sourced from starch degradation, in response to ethylene and in response to low temperature.
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PMID:Sucrose-phosphate synthase steady-state mRNA increases in ripening kiwifruit. 952 Feb 77

This study investigated the potential of restricted access media (RAM) columns used as a first column in coupled-column LC hyphenated to thermospray tandem mass spectrometry (LC/LC-TSP/MS/MS) for the fast, selective, and sensitive determination of target drugs in serum samples. Because of their wide range in polarity, salbutamol and clenbuterol were chosen as model compounds and representatives of the class of beta 2-agonists. Three types of RAM columns were tested: (i) Pinkerton ISRP (internal surface reversed phase, 5 microns), (ii) SPS (semipermeable surface, 5 microns C18), and (iii) RP-18 ADS (alkyl-diol silica, 25 microns). A 3-micron C18 column (50 mm x 4.6 mm i.d.) was chosen as the second column. Tandem mass spectrometric detection was carried out in the selected reaction monitoring (one parent-->one daughter) mode. With regard to retention and, moreover, the peak elution volume of the analytes, the ISRP material was found to perform best: a 50-mm x 4.6-mm i.d. ISRP column in combination with a 100% aqueous buffer (pH of 7.0 +/- 0.2) allowed the injection of large volumes (up to 200 microL) of sample without additional band broadening of the analytes and provided sufficient preseparation between analytes and large-molecule serum constituents. Under the selected conditions, both analytes could be determined in serum samples up to a limit of quantification (LOQ) of 0.5 ng/mL, with a sample throughput of 7 and 5 h-1 for salbutamol and clenbuterol, respectively. Method validation was carried out by analyzing, in the course of several days, calf and human serum samples spiked with the analytes. In the case of salbutamol, the overall recovery from serum samples spiked at levels between 0.5 and 50 ppb (n = 33) was 103.4%, with a repeatability of 12.7% and reproducibility of 14.3%. The overall recovery for clenbuterol was 99.6% (n = 15, spiked level 0.5-5 ppb), with a repeatability of 15.2% and reproducibility of 16.4%. The adopted LC/LC-TSP/MS/ MS analyzer appeared to be very robust under the selected conditions, and, after the period of analysis involving the processing of more than 100 mL of serum, neither loss of chromatographic performance nor pressure increase of columns or of the interface was observed.
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PMID:The potential of restricted access media columns as applied in coupled-column LC/LC-TSP/MS/MS for the high-speed determination of target compounds in serum. Application to the direct trace analysis of salbutamol and clenbuterol. 955 93

Sucrose-phosphate synthase (SPS, EC 2.4.1.14) biochemical properties and peptide composition have been analyzed in rice leaf seedlings. SPS was purified using DEAE-Sephacel chromatography, gel filtration on Sepharose 6B and anion exchange chromatography on Mono Q. At this stage two enzyme forms (SPS-I and -II) were separated. SPS-II was purified 90-fold; however, SPS-I presented a lower specific activity regarding the previous purification step and an unstable activity. Both enzyme forms had similar apparent Km values for Fru-6P but the SPS-I Km for UDP-Glc was ca. 10-fold higher than the SPS-II one. In addition, they differentiate in the capacity of being modulated by Glc-6-P and Pi: while SPS-II activity was inhibited by Pi and activated by Glc-6-P, SPS-I was not affected by either effectors. A native molecular mass of ca. 420 kDa was found by gel filtration. In SPS expression analysis using leaf rice and wheat germ SPS antibodies, a 116 kDa polypeptide was revealed in rice leaf extracts and no polypeptide was immunoactive in rice roots.
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PMID:Studies on sucrose-phosphate synthase from rice leaves. 962 Apr 36

One of the current issues in the investigation of language by means of event-related brain potentials (ERPs) is whether there is an ERP effect that can be specifically related to the processing of syntactic information. It has been claimed that a late positivity (P600 or SPS-syntactic positive shift) occurring to syntactic violations or ambiguities qualifies as such an effect. In the present investigation we compared ERPs elicited by morphosyntactic (case inflection errors), semantic, and orthographic (misspelled words) violations in a group of young German subjects. All three types of violations gave rise to late positivities having the characteristics of the previously described P600/SPS. In an earlier time window, however, semantic violations were associated with a centroparietally distributed N400 component, whereas syntactic violations gave rise to a negativity of smaller amplitude that had a frontocentral distribution. In light of the present experiment, the view that the P600/SPS as a whole reflects specific syntactic processes appears to be untenable and an alternative interpretation is proposed. The different distributions of the late positive shifts merit further investigation.
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PMID:Brain potentials and syntactic violations revisited: no evidence for specificity of the syntactic positive shift. 962 87

In the study of social anxiety, it is common to differentiate between social interaction versus performance anxiety. The Social Interaction Anxiety Scale was designed to assess social interaction anxiety, and the Social Phobia Scale to assess fear of scrutiny by others (Mattick and Clarke, 1989). In common use, these scales are typically administered together and treated as subscales of a larger measure. However, the joint factor structure of these instruments has never been examined; therefore, it is unclear whether or not the items on these scales actually represent distinct aspects of social anxiety. In the present study, a confirmatory factor analysis of the pooled items from the SIAS and SPS failed to adequately fit the data. An exploratory factor analysis yielded three factors: (1) interaction anxiety, (2) anxiety about being observed by others, and (3) fear that others will notice anxiety symptoms. However, hierarchical factor analysis suggested that these factors all load on a single higher-order factor, social anxiety. Relationships of the first-order factors to other measures of social and performance fear and avoidance are examined, and implications of our findings for the assessment of social phobia are discussed.
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PMID:Factor structure of the Social Interaction Anxiety Scale and the Social Phobia Scale. 967 Jun 4

Plant 3-hydroxy-3-methylglutaryl-CoA reductase(HMGR; EC 1.1.1.34) and sucrose-phosphate synthase (SPS; EC 2.4.1.14) and synthetic peptides designed from the known phosphorylation sites of plant HMGR (SAMS*: KSHMKYNRSTKDVK), rat acetyl-CoA carboxylase (SAMS: HMRSAMSGLHLVKRR), spinach SPS (SP2: GRRJRRISSVEJJDKK), and spinach NADH:nitrate reductase (NR6: GPTLKRTASTPFJNTTSK) were used to characterize kinase activities from cauliflower (Brassica oleracea L. ) inflorescences. The three major peaks of protein kinase activity resolved by anion-exchange FPLC are homologs of those observed previously in spinach leaves and thus are designated PKI, PKIV, and PKIII, listed in order of elution. PKIV was the most active in terms of phosphorylation and inactivation of recombinant Nicotiana HMGR and was also strictly Ca2+ dependent. The novel aspects are that PKIII has not been detected in previous cauliflower studies, that SAMS* is a more specific peptide substrate to identify potential HMGR kinases, and that the major HMGR kinase in cauliflower is Ca2+ dependent. Of the three major kinases that phosphorylated the SP2 peptide only PKI (partially Ca2+ sensitive) and PKIII (Ca2+ insensitive) inactivated native spinach leaf SPS. Cauliflower extracts contained endogenous SPS that was inactivated by endogenous kinase(s) in an ATP-dependent manner and this may be one of the substrate target proteins for PKI and/or PKIII. The substrate specificity of the three kinase peaks was studied using synthetic peptide variants of the SP2 sequence. All three kinases had a strong preference for peptides with a basic residue at P-6 (as in SP2 and SAMS*; SAMS has a free amino terminus at this position) or a Pro at P-7 (as in NR6). This requirement for certain residues at P-6 or P-7 was not recognized in earlier studies but appears to be a general requirement. In plant HMGR, a conserved His residue at P-6 is involved directly in catalysis and this may explain why substrates reduced HMGR phosphorylation in vitro.
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PMID:3-Hydroxy-3-methylglutaryl-coenzyme A reductase kinase and sucrose-phosphate synthase kinase activities in cauliflower florets: Ca2+ dependence and substrate specificities. 967 40

A sensitive and selective bioanalytical method for simultaneous determination of diclofenac and oxybuprocaine in human aqueous humor using reversed-phase HPLC and electrochemical detection is described. Chromatographic separation was achieved by using a Regis SPS 100 RP-8 column (5 microns; 150 x 4.6 mm I.D.). This support is coated with a hydrophilic polyoxyethylenepolymer. It allows protein-containing samples to be injected directly onto the column. The electrochemical detector permit a detection limit of 500 pg diclofenac per ml (daily relative standard deviation 6.3%) and 50 ng oxybuprocaine per ml (daily R.S.D. 2.6%), respectively. Results of administered and measured drug-concentrations in time dependent decrease are presented.
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PMID:Simultaneous determination of diclofenac and oxybuprocaine in human aqueous humor with HPLC and electrochemical detection. 980 Jun 54

A dual-task paradigm was used to explore the effects of cognitive load on social problem solving and autobiographical memory retrieval. The role that gender may play in mediating the relationship was also examined. Participants performed a secondary task concurrently with two primary tasks: (a) a cueing task, and (b) the Means-End Problem-Solving (MEPS) Task, during which they were required to attend to the memories retrieved during solution generation. Two dual-task conditions were employed in order that two levels of secondary task difficulty could be explored. Level of difficulty proved to be an important factor in the effects of resource reduction on the two primary tasks. Retrieval during the MEPS was effected by both the easy and difficult secondary task whereas retrieval on the cueing task was affected by the difficult task only. The results also showed that females (in contrast to males) favoured a more detailed SPS style using a specific memory database. Consequently, under central executive pressure, females' performance was significantly affected while males' performance remained largely unchanged.
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PMID:Gender differences in the dual-task effects on autobiographical memory retrieval during social problem solving. 985 6


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