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Biomedical subjects

T M Li

Publications and source records attributed to T M Li.

36 records · Page 2Linked to original sources

New sequential external counterpulsation for the treatment of acute myocardial infarction.

Over a period of 6 years, 52 patients with acute myocardial infarction (AMI) were treated with sequenced external counterpulsation (SECP). Of the 23 patients who experienced severe chest pain, 22 had complete relief within 30 min of SECP, and 31 of the 52 patients showed remarkable improvement in their electrocardiogram after the first hour of treatment. Fifteen patients were studied using the 35-lead ST segment elevation-mapping method. sigma ST and NST, indicators of infarct size, showed decreasing trends in seven patients treated with SECP for a period of 6 days, whereas sigma ST and NST of the control group of eight patients increased during the same period. Our hemodynamic data indicate that in four of five patients with AMI and left heart failure, the central venous pressure and cardiac output increased after SECP, whereas the pulmonary wedge pressure decreased. Measurements of the P-wave terminal force of lead V1 also demonstrated that the application of SECP can improve left ventricular function in a majority of patients with AMI.

Adult↗

Automated fluorometer/photometer system for homogeneous immunoassays.

A fully automated bench-top clinical analyzer (OPTIMATE TM; Ames/Gilford) performs homogeneous fluorescent immunoassays, colorimetric immunoassays, and determinations of routine blood analytes; drugs, enzymes, metabolites, specific proteins, and hormones in serum. Unique features include a combination fluorescence/absorbance aspirating thermocuvette, a photon-counting fluorometer/photometer, a multi-reagent distribution valve to dispense as many as three reagents plus buffer, and a user-replaceable programmable memory cartridge for software updates. We have evaluated the performance of OPTIMATE substrate-labeled fluorescent immunoassays for gentamicin, tobramycin, amikacin, theophylline, phenytoin, phenobarbital, primidone, carbamazepine, and quinidine with this automated system. A sample throughput of 92 samples per hour is achieved by reading fixed-point fluorescence results every 39 s after an initial 4-min reaction period. Precision studies indicate typical CVs of less than or equal to 6% for mid-range controls. Standard curves can be reused for as long as two weeks before recalibration. With clinical samples, results by the OPTIMATE procedure correlated well (r greater than or equal to 0.97) with those by a reference method.

Autoanalysis↗

Sequential external counterpulsation (SECP) in China.

A new SECP was developed and is currently successfully used in more than 120 clinics in China. This sequenced counterpulsation device has the effect of milking more blood as pressure is applied on the extremities in succession (first distally then proximally) and in decreasing values (265 to 200 mm Hg). In comparison of the 4 types of counterpulsation, namely nonsequenced leg and 4 limb counterpulsation, and sequenced 4 limb and 4 limb with buttock balloons, the SECP with buttock balloons method raised the DA to appreciably higher levels with respect to DA amplitude and area. DA was raised 43.9% higher than that obtained with 4 limb SECP alone. From our experimental and clinical results, we conclude that SECP with buttock balloons is a far more effective method providing greater diastolic augmentation than previously reported in the literature. Two hundred angina and 52 AMI patients who had undergone the SECP treatment constitute the basis of this report. The present work represents an analysis of 6 yrs experience with studies on the design, development and clinical evaluation of the SECP method. Ninety-seven percent of the angina patients obtained long-term symptomatic relief. Relapse was minimal. In 95.7% of the AMI patients chest pain or shock symptoms were rapidly relieved. SECP with high DA appears to be very effective in improving myocardial blood supply and ventricular function. Moreover, the simplicity and advantages of this noninvasive sequenced counterpulsation method have been reported (1063 cases) from 31 clinics in China. This method is convenient, safe, and far more effective than conventional drug therapy for patients with acute and chronic ischemic myocardial disease.

Angina Pectoris↗

Homogeneous substrate-labeled fluorescent immunoassay for carbamazepine.

Carbamazepine is an anticonvulsant drug useful in the management of epilepsy. Because of the narrow therapeutic range, serum carbamazepine monitoring is useful for ensuring adequate drug therapy without toxicity. We report the development of a homogeneous substrate-labeled fluorescent immunoassay for carbamazepine in human serum. A carbamazepine fluorogenic reagent (FR) has been synthesized. Upon hydrolysis by beta-galactosidase, the nonfluorescent FR produces a fluorescent product. This enzymic hydrolysis sin inhibited when the FR binds with antibody against carbamazepine. The inhibition is relieved when carbamazepine competes with FR for available antibody binding sites. Thus, increasing levels of carbamazepine result in increasing levels of fluorescence that can be conveniently monitored with any conventional fluorometer. For low, medium, and high control sera (4, 12, and 16 micrograms carbamazepine/ml), the within-run coefficient of variation for the assay is 5.5%, 1.6%, and 2.9%, respectively, while the respective between-run coefficients of variation are 3.5%, 1.9%, and 2.3%. Fifty-three clinical serum samples were assayed by the SLFIA, gas chromatography (GC), high pressure liquid chromatography (HPLC), and an enzyme immunoassay method. The SLFIA method compares favorably with the HPLC technique (r - 0.97, slope = 1.10, y-intercept = 1.21), the enzyme immunoassay (r = 0.98, slope = 1.07, y-intercept = 0.82), and the GC method (r = 0.95, slope = 1.01, y-intercept = -0.03).

Antibodies↗

Homogeneous fluorescent immunoassay with dry reagents.

We described an assay for theophylline in serum in which the reagents for the substrate-labeled fluorescent immunoassay procedure are dry reagents in a paper matrix. This assay format is both rapid and convenient. In one such paper strip assay, the competing drug conjugate is present in the diluting buffer (dilution-into-conjugate strip assay); in another, all necessary assay reagents are contained on the strip (integral strip assay). The latter strip can be prepared by using appropriate solvents in a "two-dip" procedure, which prevents premature reaction of the assay constituents. The dilution-into-conjugates strip assays yield results that compare well with those by the corresponding solution assay (r = 0.975, standard error of estimate = 1.67 mg/L) and by "high-performance" liquid chromatography (r = 0.980, standard error of estimate = 1.49 mg/L). The integral strip results were compared with those by an enzyme immunoassay for theophylline (r = 0.990, standard error of estimate = 2.07 mg/L). Both strip assays require only a single dilution step before analysis.

Humans↗

Homogeneous substrate-labeled fluorescent immunoassay for theophylline in serum.

A substrate-labeled fluorescent immunoassay for theophylline in serum is described. 8-(3-Aminopropyl)-theophylline is covalently attached to a fluorogenic enzyme substrate, 7-beta-galactosylcoumarin-3-carboxylic acid. Hydrolysis of this theophylline-labeled substrate by beta-galactosidase yields a fluorescent product. When antibody to theophylline interacts with this substrate, the resulting complex is inactive as an enzyme substrate. For measuring theophylline, competitive protein-binding reactions are set up, with the theophylline in the sample competing with the substrate for the antibody-binding sites. The substrate not bound to antibody is hydrolyzed by beta-galactosidase, producing fluorescence that is proportional to the theophylline concentration. Results for theophylline determined by this method in clinical samples of serum correlated well (r > 0.96) with results obtained by gas-chromatographic or enzyme immunoassay procedures. The within-run CV for three control samples ranged from 1.1 to 2.8%, the between-run CB from 2.3 to 4.5%.

Antibody Specificity↗

Effect of pressure upon the fluorescence of various flavodoxins.

The effects of hydrostatic pressure in the range of 10(-3) to 11 kbar on the fluorescence of flavodoxins from Peptostreptococcus elsdenii, Desulfovibrio vulgaris, Azotobacter vinelandii, and Clostridium MP were investigated. The first three flavoproteins showed under high pressure enhancements of flavin fluorescence of over 50 times resulting from the release of flavin mononucleotide from the protein complex. The Clostridial flavodoxin showed a very much smaller fluorescence change. At pH 7.5 the high-pressure fluorescence changes of the flavodoxins of D. vulgaris and P. elsdenii were not reversed by decompression, but in A. Vinelandii the pressure changes were over 80% reversible. At pH 5 over 80% reversibility was restored to the flavodoxins of D. vulgaris and P. elsdenii, although the pressure dependence of the fluorescence changes was very similar in the reversible and irreversible cases. The midpoint pressures in the reversible reactions were 4.7 kbar (D. vulgaris), 8.7 kbar (P. elsdenii), and 10.6 kbar (A. vinelandii) indicating specific differences in the flavin binding regions. Apparent volume changes in these reactions were 65-75 mL/mol indicating participation of a large fraction of the protein in the pressure-induced changes. The irreversible changes are not related to protein aggregation and are believed to result from a pressure-dependent covalent modification, not yet characterized, of the flavin binding region of the protein.

Azotobacter↗

Volume changes in the formation of internal complexes of flavinyltryptophan peptides.

The effect of pressure, up to 10 kbar, on the fluorescence yield and lifetime of two flavinyltryptophan peptides was investigated. These peptides differed only in the number of methylene groups, respectively three and five, separating the chromophores. At atmospheric pressure the closed nonfluorescent form predominated in both compounds constitutin 94% of the total in the short-linked peptide and 80% in the long-linked one. The fluorescence of both peptides decreased at high pressure and the volume change upon formation of the nonfluorescent complex in the short peptide (--1.8 mL/mol) was less than half of the change in the long peptide (--4.8 mL/mol) or the value for FAD (--4.3 mL/mol). The much smaller compressibility of the short peptides is attributed to the mechanical constraint to the approach of the interacting rings, imposed by the short link. Mechanical constraints of similar nature may be expected to be operative in proteins. Their importance in pressure denaturation is discussed.

Flavoproteins↗

Effects of pressure upon the fluorescence of the riboflavin binding protein and its flavin mononucleotide complex.

The effect of pressure in the range of 10(-3)-10 kbars upon the ultraviolet fluorescence of the riboflavin binding protein and the fluorescences of its complex with flavin mononucleotide has been studied. The fluorescence spectrum of the isolated protein showed a reversible red shift of 12nm (1000 cm-1) at high pressure, indicating the reversible exposure of the tryptophan to solvent. From the pressure dependence of the visible fluorescence of the protein-flavin complex in the region of 1-4 kbars the volume change in dissociation of the protein-ligand complex was estimated to be +3.3ml/mol. A very sharp increase in fluorescence-up to 30-fold of the low-pressure value-takes place in the region 5-8 kbars. This increase is due to release of the flavin from the complex and is assigned to pressure denaturation of the protein. The midpoint, rho 1/2, of this transition was found at 6.5 kbars and the change in volume, delta, in the reaction (native-to-denatured) was calculated to be -74ml/mol. Addition of up to 30% methanol results in a progressive decrease both in delta and rho 1/2, in agreement with the concept that hydrophobic bonding stabilizes the native structure.

Bromosuccinimide↗

Performance of a fully automated in vitro allergy testing system.

BACKGROUND: Since the development of the radioallergosorbent test (RAST) for quantification of allergen-specific IgE, numerous non-radoisotopic methods have been devised which combine the proven cellulose disc technology with enzyme-linked immunoassay methods. The HY.TEC EIA (Hycor Biomedical, Inc. Irvine, CA) was compared with Pharmacia CAP with respect to overall system features and assay performance characteristics. METHODS: The HY.TEC EIA and Pharmacia CAP were compared with respect to calibrator range, sensitivity, type of detection, type of solid phase, throughput, and mode of operation. To determine the assay sensitivity and specificity for a variety of allergens, a total of 2,447 tests were performed on both CAP and HY.TEC EIA. The samples were scored positive in both cases using a cutoff of 0.35 IU/mL. RESULTS: The general features of the HY.TEC EIA system are comparable to Pharmacia UniCAP, with the added advantage of higher throughput. Intra-assay precision was 7% and inter-assay precision was 9-15%. Using CAP as a comparative method, HY.TEC EIA has a sensitivity of 94.0% and a specificity of 94.4%. CONCLUSIONS: The HY.TEC EIA demonstrates excellent agreement with the Pharmacia CAP system in the determination of allergen-specific IgE. With the automation necessary in today's clinical laboratory, we conclude that the HY.TEC EIA is a state-of-the-art tool for the diagnosis of allergic disease.

Allergens↗