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Biomedical subjects

L Sun

Publications and source records attributed to L Sun.

At least 541 records · Page 30Linked to original sources

Comparison of enzyme immunoassay and gas chromatography for determination of carbamazepine and ethosuximide in human serum.

Patients' sera were analyzed for carbamazepine and ethosuximide by enzyme immunoassay (x) and gas chromatography (y), and the results were compared. The correlation coefficients were: for carbamazepine, x vs. y 0.94 (n = 93); for ethosuximide, x vs. y 0.99 (n = 30). These results suggest that the two methods could be used interchangeably. Ten serum samples supplemented with carbamazepine (2.5 to 12.5 mg/liter) and ethosuximide (20.0 to 130.0 mg/liter) were analyzed by both methods. The correlation coefficients were: x vs. y 0.99 (n = 10) for carbamazepine and x vs. y 0.99 (n =10) for ethosuximide.

Carbamazepine↗

Radioimmunoassay, enzyme immunoassay, spectrophotometry, and gas-liquid chromatography compared for determination of phenobarbital and diphenylhydantoin.

Sera from epileptic patients were assayed for phenobarbital and diphenylhydantoin by four different analytical procedures. Quantitative results obtained by radioimmunoassay (I) and enzyme immunoassay (II) were compared to each other and to the results obtained on aliquots of the same sample by gas-liquid chromatography (III) and ultraviolet spectrophotometry (IV). For phenobarbital the correlation coefficients were I vs. II, 0.909; I vs. III, 0.947; II vs. III, 0.917; I vs. IV, 0.950; II vs. IV, 0.953. For diphenylhydantoin the correlation coefficients were I vs. II, 0.953; I vs. III, 0.951; II vs. III, 0.957; I vs. IV, 0.862; II vs. IV, 0.898. The immunoassays can be substituted for liquid chromatography or ultraviolet spectrophotometry without changing the resulting clinical interpretations.

Chromatography, Gas↗

Primidone analyses: correlation of gas-chromatographic assay with enzyme immunoassay.

Serum specimens from patients on primidone therapy were assayed by two currently available procedures: a commercially available enzyme immunoassay (EMIT) and a published gas-chromatographic procedure. Both procedures were done with commercially available materials and equipment. Results by the two procedures agreed well, which suggests that the two methods could be used interchangeably. For the 94 specimens studied, the correlation coefficient was 0.98, and the least-squares values of slope and intercept were, respectively, 0.97 and 0.51 mg/liter.

Chromatography, Gas↗

Radioimmunoassay and enzyme immunoassay compared for determination of digoxin.

Patients' sera were analyzed for digoxin by using two different radioimmunoassays and an enzyme immunoassay. Quantitative results obtained by enzyme immunoassay (I) were compared to results obtained on aliquots of the same sample by the radioimmunoassays (II and III). The correlation coefficients were: I vs. II 0.90, n=108; I vs. III 0.94, n=102; and II vs. III 0.95, n=158. Day-to-day precision (10 days) on a low control (1.3 mug/liter) and a high control 3.0 mg/liter), expressed as coefficients of variation, were: I, 13% and 7.8%, II, 4.0% and 4.7%; and III, 8.9% and 4.2%. Ten digoxin-supplemented samples (0-8 mug/liter) were analyzed by the three methods. Correlation coefficients were: supplemented sample vs. I, O.99; supplemented sample vs. II, 0.97; supplemented sample vs. III, 0.98.

Digoxin↗

The specificity of different classes of ethylating agents toward various sites of HeLa cell DNA in vitro and in vivo.

The sites and extent of ethyl products of neutral ethylation of HeLa cell DNA by [14-C]diethyl sulfate, [14-C]ethyl methanesulfonate, and [14-C]ethylnitrosourea have been determined in vitro and in vivo, and found to differ significantly depending on the ethylating agents. Diethyl sulfate and ethyl methanesulfonate ethylate the bases of HeLa cell DNA in the following order: 7-ethylguanine greater than 3-ethyladenine greater than 1-ethyladenine, 7-ethyladenine greater than 3-ethylguanine, 3-ethylcytosine, O-6-ethylguanine. Ethyl bases accounted for 84-87% of the total ethyl groups associated with HeLa cell DNA. Ethylnitrosourea, in contrast, has particular affinity for the O-6 position of guanine. It ethylates the bases of HeLa cell DNA in the following order: O-6-ethylguanine, 7-ethylguanine greater than 3-ethyladenine greater than 3-ethylguanine, 3-ethylthymine greater than 1-ethyladenine, 7-ethyladenine, 3-ethylcytosine. Ethylation of the bases only accounts for 30% of the total ethylation in the case of ethylnitrosourea. The remaining 70% of the [14-C]ethyl groups, introduced in vivo and in vitro, are in the form of phosphotriesters which after perchloric acid hydrolysis are found as [14-CA1ethanol and [14-C]ethyl phosphate. In contrast, phosphotriesters amounted to only 8-20% of total ethylation in in vivo or in vitro diethyl sulfate and ethyl methanesulfonate treated HeLa cell DNA, and 25% of the total methylation in in vitro methylnitrosourea treated HeLa cell DNA. Alkylation at the N-7 and N-3 positions of purines in DNA destabilizes the glycosidic linkages. Part of 7-ethylguanine and 3-ethyladenine are found to be spontaneously released during the ethylation reaction. Incorporation of the 14-C of the alkylating agents into normal DNA bases of HeLa cells can be eliminated by performing the alkylations, in the presence of cytosine arabinoside, for 1 hr.

Binding Sites↗

Effects of alkylation of phosphodiesters and of bases of infectivity and stability of tobacco mosaic virus RNA.

Upon ethyl nitrosourea treatment of RNA of tobacco mosaic virus, up to four phosphodiester groups may be alkylated per molecule without chain breakage, as shown be sucrose gradient centrifugation. This indicates that ribophosphotriesters are quite stable. However, when this alkylation reaction is of longer duration and 6 to 10 triesters are formed, then an average of 1 to 2 breaks occurs and little or no intact RNA can be isolated. Methyl nitrosourea is less effective in forming triesters (about 25% of total alkyl groups compared to about 65% for ethyl nitrosourea), and a greater number of alkyl groups can, therefore, be introduced before breaks occur. Diethyl sulfate and dimethyl sulfate, which alkylate almost only the bases of nucleic acids, do not cause significant degradation of RNA of tobacco mosaic virus, even when as many as 70 alkyl groups are bound. All types of alkylation cause similar losses in viral infectivity at low levels of alkylation. Thus, an average of two chemical events leads to one lethal event, regardless of the nature of the alkylating reaction, which, for example, is with dimethyl sulfate about 65% on the N-7 guanine while with ethyl nitrosourea it is about 65% on phosphodiesters. It is thus concluded that all alkyl groups, whether on the base or on the phosphate, have the same potential to cause inactivation and that inactivation of RNA can result from phosphotriester formation per se.

Alkylation↗

A longitudinal study of emergency room visits and air pollution for Prince George, British Columbia.

A new method for regression analysis of longitudinal counts is applied to data from Prince George, British Columbia, previously analysed by Knight et al. The data consist of daily recordings of the number of emergency room visits for each of four categories of respiratory diseases, along with measurements of meteorological variables and air pollution. We use a state-space model assuming conditionally independent Poisson counts for the four categories given a latent morbidity process, the latent process being a gamma Markov process. The main objective of the investigation was to examine the relationship between air pollution and respiratory morbidity, taking into account seasonality and meteorological conditions. We found that total reduced sulphur significantly influences the expected number of emergency room visits for the four disease categories, in agreement with the conclusion by Knight et al. However, our final model is simpler than theirs; in particular we found no evidence of seasonal variation beyond that explained by the meteorological variables.

Air Pollution↗

Synthesis and terminal functionalization of a polymerizable phosphatidylethanolamine.

We report the design and synthesis of bifunctional phospholipid conjugates, which contain a polymerizable acrylate group and a terminal linker, such as biotin or N-(epsilon-maleimidocaproyl (EMC), to facilitate bioconjugation reactions. The lipid conjugate can be used to generate a multifunctional substrate-supported phospholipid film that is further stabilized via in-situ photocopolymerization.

Acrylates↗

5'-sulfhydryl-modified RNA: initiator synthesis, in vitro transcription, and enzymatic incorporation.

The detailed syntheses of the sulfhydryl-modified guanosine monophosphates 5'-deoxy-5'-thioguanosine-5'-monophosphorothioate (GSMP), O-[omega-sulfhydryl-tetra(ethylene glycol)]-O-(5'-guanosine) monophosphate (5'-HS-PEG4-GMP), and O-[omega-sulfhydryl-di(ethylene glycol)]-O-(5'-guanosine) monophosphate (5'-HS-PEG2-GMP) are reported. Transcription reactions employing GSMP, 5'-HS-PEG4-GMP, or 5'-HS-PEG2-GMP as the initiator nucleotide for T7 RNA polymerase introduce a thiol group at the 5'-end of RNA. The efficiency of thiol incorporation at the 5'-terminus of modified RNA compounds was assayed with three different thiol-reactive biotinylated reagents followed by streptavidin gel-shift methods. The transcription efficiency with various ratios of GTP to 5'-HS-PEG2-GMP was explored by reaction with a sulfhydryl-reactive maleimide-conjugated protein. This is an efficient method to incorporate enzymatically a thiol group into the 5'-end of RNA.

Biotinylation↗

Neoglycophospholipids with alkyl spacers: synthesis via an improved reductive amination and monolayer properties.

An efficient synthesis of neoglycophospholipids with variable length alkyl spacer chains is described. Neoglycophospholipids tethered by alkyl chains of 3, 5, 7, 10, and 16 methylene units were synthesized in good overall yields in four steps. The key intermediates, omega-oxoalkyl glycopyranosides, were synthesized in two steps by glycosidation of chloro (or ethylthio) glycosides with a diol followed by oxidation of the remaining hydroxy group to an aldehyde functionality. Conjugation of the omega-glycoalkyl aldehyde with distearoylphosphatidylethanolamine via an improved reductive amination procedure significantly enhanced efficiency and yields with respect to those from traditional procedures. The amphiphilic properties of the neoglycophospholipids were characterized at the air-water interface. While the carbohydrate head group had relatively little effect, the length of the alkyl spacer profoundly influenced surface area-pressure isotherms.

Amines↗

Flow through, immunomagnetic cell separation.

A brief, process-oriented overview of immunologically based cell separation technology is presented. In addition, the design and preliminary experimental data of two unique flow-through immunomagnetic cell separation devices are presented. The first design is based on a dipole magnetic field, while the second design is basis on a quadrupole magnetic field. The dipole design can "fractionate" an inlet, magnetically labeled, cell stream into different outlet streams on the basis of the degree to which the cell is immunomagnetically labeled. The quadrupole separator splits an inlet, immunomagnetically labeled, cell stream into two outlet streams in which the purity, recovery, and potentially the degree to which the cells are immunomagnetically labeled is controlled by the flow rates in the inlet and outlet flows. A 99% purity and 86% recovery have been achieved with this system. Some distinct advantages of these two systems are the potential of high purity, recovery, and throughput at a cost which is potentially significantly lower than current, comparable technologies.

Electrochemistry↗

Uniform band intensities in fluorescent dye terminator sequencing.

The use of Cyanine dye (Cy5 and Cy5.5) labeled dideoxy terminators with Thermo Sequenase DNA polymerase in DNA sequencing provides uniform band intensity, improved sequence read-length, and accuracy. It also greatly improves the ability to detect single base heterozygotes with dye-terminator sequencing method.

Base Sequence↗