[Establishing indicators for diagnosis of cholinergic crisis].

Cholinergic crisis is an adverse effect of an anticholinesterase agent, which is one of the cholinergic agents. Cholinergic crisis may induce serious conditions such as breathing difficulties. Cholinergic crisis is often diagnosed by an abnormally low level of serum cholinesterase (ChE). However, ChE value is not an appropriate indicator of cholinergic crisis since it has a high inter-individual variation, even though its intra-individual variation is low. Therefore, an indicator with less inter-individual variation capable of preventing the risk of cholinergic crisis was investigated. The results of correlation test between ChE and serum albumin (Alb) showed a strong positive correlation; r = 0.778 in BCG method(Bromo cresol green method) and r = 0.766 in the BCP-improved method for Alb. In addition, the variations of Alb values are much lower than the drastic depression of ChE values in cholinergic crisis. Thus, it is considered that the ratio of ChE and Alb (ChE/Alb) can be a useful indicator of cholinergic crisis. As a result of ROC (Receiver operating characteristic) analysis, the ratios of ChE and Alb values using the BCG method (ChE/Alb (BCG)) were 20.7, 87.0, 156.8 for the Cutoff value, Likelihood ratio and Odds ratio respectively. When using the BCP-improved method for Alb, the ratios of ChE and Alb (ChE/Alb(BCP improved)) were 25.0, 93.7, 180.1 for the Cutoff value, Likelihood ratio and Odds ratio respectively. The ChE/Alb ratio appears to be an excellent indicator of cholinergic crisis diagnosis since it shows a high likelihood ratio as well as a high odds ratio.

[Urinary diacetylspermine: its analysis and performance as a novel tumor marker].

N1,N12-Diacetylspermine (DiAcSpm) is excreted in the urine of healthy persons as a minor component of urinary polyamine. It is a promising tumor marker, since its excretion is frequently elevated in patients with various types of cancers. DiAcSpm was first detected and characterized by HPLC fractionation followed by enzymatic detection, but more recently, antibodies highly specific for DiAcSpm was prepared, and an ELISA system applicable to determination of urinary DiAcSpm was established. Measurement of urinary DiAcSpm using this ELISA system revealed that DiAcSpm is able to detect early stage (m and sm) colon cancers which CEA and CA19-9 cannot detect. DiAcSpm may also serve as a prognostic indicator and a marker for recurrence of colon cancer. Urinary DiAcSpm is elevated in metastatic and primary brain tumors including grade 3 and 4 gliomas and primary central nervous system lymphoma. In these primary brain tumors changes in urinary DiAcSpm were well correlated with the efficacy of treatments, recurrence of disease and increased malignancy of a tumor. DiAcSpm may be useful as a comprehensive indicator of the activeness of a brain tumor lesion in a patient. DiAcSpm was elevated in hepatocellular carcinoma, but patients with liver cirrhosis also showed considerably elevated levels of DiAcSpm.

Development and application of serum cholinesterase activity measurement using benzoylthiocholine iodide.

BACKGROUND: Measurement of cholinesterase activity in serum is important to identify substantial liver disease and damage by pesticides, and to assess the degree of development of fatty liver and preoperative risk. Many procedures using various artificial substrates have been developed but suffer from problems with substrate specificity and interference by endogenous substances. METHODS: An assay pseudocholinesterase (ChE, EC 3.1.1.8 acylcholine acylhydrolase) activity was developed using a stable substrate specific to ChE, benzoylthiocholine iodide (BZTC). The thiocholine generated by hydrolysis of BZTC by ChE activity reacts with 2, 2'-dipyridildisulfide (2-PDS) to produce 2-thiopyridine (2-TP), which is measured at 340 nm. Optimum pH, buffer types and concentrations, substrate concentrations, and optimum conditions of the color reaction were investigated. The substrate specificity, test interferences, correlation with other measurement methods, and reference interval were evaluated. RESULTS: The optimum pH of this method was 7.8, and 3-[4-(2-hydroxyethyl)-1-piperazinyl] propanesulfonic acid (EPPS) buffer solution was selected. Constant activity was shown at buffer concentrations >200 mmol/l, and the maximum activity was shown at a substrate concentration of 0.2 mmol/l. When a Hill plot was utilized, the Hill number was 1.08 and 1.09. The reaction velocity at this substrate concentration was 94% of V(max). The K(m) of ChE to BZTC was between 1.2 x 10(-2) and 1.3 x 10(-2) mmol/l. The range was 0-300 U/l. The coefficients of variation (CV) for 20 measurements of serum containing 53.1, 96.6, and 270.7 U/l of ChE were 0.82%, 0.76%, and 0.54%, respectively. The relative reactivity of acetylcholinesterase (AChE) to this substrate was 2%. The correlation factors of this method to three other methods were between 0.993 and 0.998. CONCLUSIONS: This method provides excellent specificity, reproducibility, a wide measurement range, and minimal interference from endogenous substances to common serum analytes. Correlation of this method with conventional methods was good. Because the reagents are stable after preparation, this assay is useful for routine analysis.