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S Kurooka

Publications and source records attributed to S Kurooka.

At least 19 recordsLinked to original sources
PubMed1990

A sandwich-enzyme immunoassay for the quantification of lipoprotein lipase and hepatic triglyceride lipase in human postheparin plasma using monoclonal antibodies to the corresponding enzymes.

We have developed a sandwich-enzyme immunoassay (EIA) for the quantification of lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL) in human postheparin plasma (PHP) using monoclonal antibodies (MAbs) directed against the corresponding enzymes purified from human PHP. The sandwich-EIA for LPL was performed by using the combination of two distinct types of anti-LPL MAbs that recognize different epitopes on the LPL molecule. The immunoreactive mass of LPL was specifically measured using a beta-galactosidase-labeled anti-LPL MAb as an enzyme-linked MAb, an anti-LPL MAb linked with the bacterial cell wall as an insolubilized MAb, and purified human PHP-LPL as a standard. The sandwich-EIA for HTGL was carried out by using two distinct anti-HTGL MAbs that recognize different epitopes on HTGL. The limit of detection was 20 ng/ml for LPL and 60 ng/ml for HTGL. Each method yielded a coefficient of variation of less than 6% in intra- and inter-assays, and a high concentration of triglyceride did not interfere with the assays. The average recovery of purified human PHP-LPL and -HTGL added to human PHP samples was 98.8% and 97.5%, respectively. The immunoreactive masses of LPL and HTGL in PHP samples, obtained at a heparin dose of 30 IU/kg, from 34 normolipidemic and 20 hypertriglyceridemic subjects were quantified by the sandwich-EIA. To assess the reliability of the measured mass values, they were compared with the corresponding enzyme activities measured by selective immunoinactivation assay using rabbit anti-human PHP-LPL and -HTGL polyclonal antisera. Both assay methods yielded a highly significant correlation in either normolipidemic (r = 0.945 for LPL; r = 0.932 for HTGL) or hypertriglyceridemic subjects (r = 0.989 for LPL; r = 0.954 for HTGL). The normal mean (+/- SD) level of lipoprotein lipase mass and activity in postheparin plasma was 223 +/- 66 ng/ml and 10.1 +/- 2.9 mumol/h per ml, and that of hepatic triglyceride lipase mass and activity was 1456 +/- 469 ng/ml and 26.4 +/- 8.7 mumol/h per ml, respectively. The present sandwich-enzyme immunoassay methods make it possible to study the molecular nature of LPL and HTGL in PHP from patients with either primary or secondary hyperlipoproteinemia.

Adult↗
PubMed1990

Competitive binding enzyme immunoassay for zonisamide, a new antiepileptic drug, with selected paired-enzyme labeled antigen and antibody.

We assessed the competitive binding between zonisamide (ZNS) in serum samples and beta-galactosidase-labeled ZNS derivatives, using competing antibodies to ZNS derivatives, and selected the best enzyme-labeled antigen and antibody for accurate enzyme immunoassay (EIA) of ZNS in serum without interference from its metabolites or from other antiepileptic drugs. This EIA, based on use of antibody linked to bacterial cell walls, has advantages over HPLC in simplicity, speed (50 samples per hour), and lack of requirement for special equipment. The concentrations of ZNS in serum as measured by the EIA correlated well with those by HPLC (n = 33, r = 0.977).

Antibodies↗
PubMed1989

Differential determination of recombinant human interleukin-1 alpha and its deamidated derivative by two sandwich enzyme immunoassays using monoclonal antibodies. Comparison with a polyclonal antibody-based competitive enzyme immunoassay.

Using three different monoclonal antibodies (McAb no. 374, no. 964 and no. 1190) to human interleukin-1 alpha (rHu-IL-1 alpha), we have established two sandwich enzyme immunoassays (EIA) to differentiate rHu-IL-1 alpha and its deamidated derivative (rHu-Asp36-IL-1 alpha) where the asparagine at position 36 (counting from the N-terminus) of rHu-IL-1 alpha is converted to Asp. The McAb no. 1190 reacts specifically with rHu-IL-alpha and not with the rHu-Asp36-IL-1 alpha whereas both no. 374 and no. 964 can react with the two different forms of rHu-IL-1 alpha. The first EIA (S-EIA I) which uses the McAb no. 964 labelled with horse-radish peroxidase and the McAb no. 1190 fixed to the microtiter plate, only measure rHu-IL-1 alpha. The second EIA (S-EIA II) which uses enzyme labelled no. 964 and no. 374 fixed to the plate, can detect both rHu-IL-1 alpha and rHu-Asp36-IL-1 alpha and this assay of total rHu-IL-1 alpha is comparable to a competitive EIA using an enzyme-labelled rHu-IL-1 alpha and an anti-rHu-IL-1 alpha polyclonal antibody. Thus, the level of rHu-Asp36-IL-1 alpha in the samples containing the two IL-1 alpha s can be calculated by subtracting the level measured by S-EIA I from that measured by S-EIA II. The two EIA systems with an assay range of 1.5-100 ng/ml do not recognize IL-1 beta, IL-2, rHu-TNF alpha, IFN-alpha and IFN-gamma of human origin.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Simple enzyme immunoassay methods for recombinant human tumor necrosis factor alpha and its antibodies using a bacterial cell wall carrier.

We have developed simple methods for measuring recombinant human tumor necrosis factor alpha (rHu-TNF alpha) and antibodies to rHu-TNF alpha in the sera of animals intravenously injected with rHu-TNF alpha. rHu-TNF alpha was measured by a competitive binding enzyme immunoassay (C-EIA) using standard rHu-TNF alpha, beta-galactosidase labeled rHu-TNF alpha as enzyme-labeled antigen (E-Ag) and anti-rabbit IgG goat immunoglobulins coupled to bacterial cell walls (insolubilized second antibody). In contrast, anti-rHu-TNF alpha antibodies were measured by a sandwich EIA (S-EIA) using purified anti-rHu-TNF alpha rabbit IgG as standard, beta-galactosidase labeled rHu-TNF alpha as E-Ag, and rHu-TNF alpha coupled to bacterial cell walls as insolubilized antigen. C-EIA permits the determination of serum rHu-TNF alpha within the range of 2-150 U/ml (about 0.7-52 ng/ml) with a CV of below 7.6% and 99% recovery. S-EIA permits the determination of anti-rHu-TNF alpha antibodies within the range of 70-1000 ng/ml with a CV of less than 4% and 94.8-106.9% recovery.

Animals↗
PubMed1987

Role of bradykinin generating and degrading systems in the vascular permeability response induced with kaolin in rats.

We investigated how bradykinin mediates inflammatory reactions in rats, via measurements of bradykinin by enzyme immunoassay method in inflammatory tissue fluids. Vascular permeability was increased markedly during the first 10 min and then declined quickly after the infusion of a kaolin suspension (10 mg/ml) in 0.8% carboxymethl-cellulose solution into an air pouch formed on the back of rats. Bradykinin in the exudate reached a maximum 5 min after the challenge and then decreased quickly. Local treatment with DL-2-mercaptomethyl-3-guanidinoethylthiopropanoic acid, an inhibitor of kininase I, and captopril, an inhibitor of kininase II in the first 10-min period, each enhanced the vascular permeability increase accompanied by the elevation of the bradykinin level, whereas soybean trypsin inhibitor, a plasma kallikrein inhibitor, lowered both vascular permeability and bradykinin. When applied in the period of 3.5 to 4 hr after the challenge, only the kininase II inhibitor was effective in elevating both vascular permeability and the bradykinin level, whereas soybean trypsin inhibitor was ineffective on vascular permeability. A bradykinin-degrading activity appeared in the exudate as early as 10 min after the challenge. These results suggest that bradykinin plays an essential role for the sudden rise of the vascular permeability observed immediately after the infusion of kaolin suspension. In the later stage (3.5-4 hr), bradykinin level remained below the assay limit of 0.07 ng/ml in spite of its active generation, presumably because of its rapid degradation by the kininases, although it still played a definite role in the vascular permeability increase.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed1987

Differential assay of salivary and pancreatic alpha-amylase in serum and urine, with use of monoclonal antibody to human salivary amylase immobilized on bacterial cell wall.

We previously reported (Clin Chim Acta 1986;159:89) that bacterial cell wall chemically coated with a monoclonal antibody specific to human salivary (S) amylase (EC 3.2.1.1) could be successfully used to separate S and pancreatic (P) amylase in solution. We have now applied this method to serum and urine samples and found that the activities of S and P amylases so measured correlated well with those measured by the isoamylase inhibitor method. The present method is simple and reliable for routine clinical tests.

Antibodies, Monoclonal↗
PubMed1984

Serum lipase response to cerulein secretin stimulation test in patients with pancreas-associated diseases as measured by sensitive colorimetric assay (a BALB-DTNB method).

We have compared responsiveness of serum lipase and amylase activity to the pancreatic exocrine stimulation with cerulein and secretin (CS test) in normal subjects and patients with pancreas-related and other diseases. The lipase and amylase activities were measured by a sensitive colorimetric method, the BALB-DTNB method and the Caraway method, respectively. The percentage of positive lipase and amylase response cases was as follows: confirmed chronic pancreatitis (N = 22), 27 and 14%; suspected chronic pancreatitis (N = 37), 46 and 32%; pancreatic cancer (N = 16), 44 and 25%; biliary tract diseases (N = 11), 14 and 14%; miscellaneous (N = 11), 0 and 18%; normal subjects (N = 13), and partial pancreatectomy (N = 5), 0 and 0%, respectively. The serum lipase response cannot be regarded as specific for pancreatic diseases because the lipase response cases were found in biliary tract diseases as well. However, in view of frequent, fast, and intense responsiveness to the CS test, the serum lipase activity measured by the BALB-DTNB method may be more useful than the serum amylase as an auxiliary diagnostic aid for suspected pancreatitis which might develop into confirmed chronic pancreatitis or cancer of the head or body of the pancreas.

Adolescent↗
PubMed1983

Semi-automated continuous-flow enzyme immunoassay for antiepileptic drugs in serum.

We have developed a semi-automated method for measuring five kinds of antiepileptic drugs in serum by successfully adapting commercial competitive-binding enzyme immunoassay kits (MARKIT; Dainippon) for use with a continuous-flow analyzer (Technicon AutoAnalyzer II equipped with a dialyzer). The free enzyme-labeled drug is automatically separated by a microfilter from the competitive immunoreaction mixture between labeled and unlabeled drug for anti-drug immunoglobulin coupled to bacterial cell walls. The concentrations of the antiepileptic drugs in serum samples can be determined by automated measurement of enzyme activity of the enzyme-labeled drugs. Results of the semi-automated method correlated well with those obtained by manual enzyme immunoassay, gas-liquid chromatography, and "high-pressure" liquid chromatography. The correlation coefficients were all greater than 0.95, showing the practicality of this method for therapeutic monitoring of antiepileptic drugs.

Anticonvulsants↗
PubMed1982

Assays of serum lipase by the "BALB-DTNB method" mechanized for use with discrete and continuous-flow analyzers.

We successfully adapted the dimercaprol (BAL) tributyrate-5,5'-dithiobis(2-nitrobenzoic acid) method (J. Biochem. 81: 361, 1977) for assay of lipase in human serum to a discrete analyzer (the TBA 880) (I) or a continuous-flow analyzer (AutoAnalyzer, Type II) (II). In both, BAL-tributyrate is used as substrate, in combination with serum esterase inhibitors and a chromogenic reagent for the SH group of the liberated BAL. Serum lipase activities of patients with pancreatic diseases, measured at 90 or 40 samples per hour by I or II, respectively, correlated well with those measured by the corresponding manual method or by Kaplan's radioassay (Anal. Biochem. 33: 213, 1970). The correlation coefficients were all greater than 0.95, and the coefficients of variation were less than 8%, showing the practical usefulness of these procedures.

Autoanalysis↗
PubMed1978

Properties of serum lipase in patients with various pancreatic diseases. Analysis by a new serum lipase assay method (the BALB-DTNB method) in combination with gel-filtration and iso-electrofocusing techniques.

Very low levels of lipase can easily be measured by a new serum lipase assay method (the BALB-DTNB method), using BAL-tributyrate (BALB) as a substrate, 5,5'-dithiobis(2-nitrobenzoic acid) as a chromogenic SH reagent, phenylmethylsulfonylfluoride as an inhibitor of esterases and sodium dodecyl sulfate as a surfactant. The BALB-DTNB method has a higher sensitivity than the conventional serum lipase assay methods, and proved useful for analyzing the properties of serum lipases in combination with gel-filtration on a Sephacryl S 200 column and isoelectrofocusing in an Ampholine column. Serum samples containing high levels of lipases from patients with pancreatic diseases or patients in whom the pancreatic exocrine gland had been stimulated by injecting caerulein and secretin were analyzed by these methods. The lipolytic profiles obtained indicated the presence of a lipase with an estimated molecular weight of 46,000 and isoelectric points of 7.4, 6.8, or/and 6.4. A lipase with properties similar to those of the serum lipase was found to be present in human pancreatic juice.

Chromatography, Gel↗
PubMed1977

A novel and simple colorimetric assay for human serum lipase.

A new and simple colorimetric method for human serum lipase [EC 3.1.1.3] assay has been developed, using 2,3-dimercaptopropan-1-ol tributyroate as a substrate, 5,5'-dithiobis(2-nitro-benzoic acid) as a chromogenic reagent, phenylmethylsulfonyl fluoride as an inhibitor of serum esterases, and sodium dodecylsulfate as a lipase activator. The method requires only 50 micron1X2 of serum sample and a reaction time of less than 30 min. The method is reproducible and sensitive enough to measure low levels of lipase activity in normal and abnormal sera. The gel filtration of serum samples on a Sephadex G-200 column gave one peak of lipase activity, when measured by the present method, and the molecular weight of the enzyme was identical with that of lipase of human pancreatic origin, confirming the specificity of this new method for the serum lipase.

Chromatography, Gel↗
PubMed1976

Relationship between the structures of S-acyl thiol compounds and their rates of hydrolysis by pancreatic lipase and hepatic carboxylic esterase.

About 100 S-fatty acyl thiol compounds designed as substrates for pancreatic lipase [EC 3.1.1.3] were synthesized and tested for susceptibility to hydrolysis by hog pancreatic lipase and hog hepatic carboxylic esterase [EC 3.1.1.1] using 5,5'-dithiobis(2-nitrobenzoic acid) as a chromogenic reagent to determine the hydrolytic rates of their S-acyl bonds. In general, the hydrolytic rates of S-acyl bonds by the lipase were fast with thioglycerol type thiol moieties, slow with dithioethyleneglycol type or monothiol type, and negligible with thiopolyol type. As for the acyl moieties, the hydrolysis of the S-acyl bonds was fast with C3-5 acyl groups, followed by C6-8 acyl groups, and the rate decreased as the acyl chain length deviated from these values or branched. On the other hand, the hydrolysis of S-acyl bonds by the esterase occurred with all types of S-acyl esters except for esters of long S-acyl chains. Of all the compounds tested with the lipase, the rate of hydrolysis of S-acyl bond was maximum with 2,3-dimercaptopropan-1-ol tributyroate [I], high with 3-mercaptopropane-1,2-diol tributyroate [II], but negligible with the analogous compound, 1,3-dimercaptopropan-2-ol tributyroate. Compounds [I] and [II] may be practically useful as substrates for lipase assay in human serum samples pretreated with phenylmethylsulfonyl fluoride, a potent inhibitor against both serum arylesterase [EC 3.1.1.2] and hepatic esterase, which attack [I] and [II].

Alcohols↗