We have developed a fully enzymatic method to measure 1,5 anhydro-D-glucitol (1,5-AG) in serum through use of pyranose oxidase (PROD: EC 1.1.3.10), glucokinase (EC 2.7.1.2), and an ATP-regenerating system. In a previous report (Clin Chem 1989;35:2039-43) the glucose interfering with the measurement of 1,5-AG was removed with a minicolumn. In the method used here, glucokinase and an ATP-regenerating system efficiently convert glucose to the unreactive compound, glucose 6-phosphate, making the method selective for 1,5-AG. The hydrogen peroxide produced in the oxidation of 1,5-AG by PROD is detected with a standard enzymatic color-developing system. The within-run and day-to-day precision (CV) of this method was 0.52-1.29% and 1.17-4.48%, respectively. The correlation (r) between the results obtained with our proposed method (y) and those obtained with the mini-column method (x) was 0.998 (y = 1.007x + 0.493 mg/L; n = 100; Sy/x = 0.641 mg/L). This newly developed method allows quicker and easier measurement of serum 1,5-AG than previously described methods.

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A simple high performance liquid chromatographic method combined with an enzyme sensor has been developed to measure 1,5-anhydroglucitol in urine. The enzyme sensor consists of a hydrogen peroxide electrode and a chitosan membrane of an immobilized pyranose oxidase. As the system does not resist interfering substances, urine samples are first purified by passing them through a two-layer column packed with (1) strongly basic anion (OH- form, the upper layer) and (2) strongly acidic cation (H+ form, the lower layer) exchange resins. 1,5-Anhydroglucitol is efficiently recovered in the flow-through fraction of the column. In this system, the minimum detectable concentration of 1,5-anhydroglucitol is 0.1 mg/L, and the measurable range extends from 0.1 to 60 mg/L. The coefficient of variation values of the within-day and day-to-day precisions are 3.0-6.5% and and 4.4-6.7% respectively, and there is good agreement between the results measured by our method and those obtained by the gas-liquid chromatographic/mass spectrometric method (r = 0.994). The method we have described here has been successfully used to elucidate a mechanism for the reducing 1,5-anhydroglucitol level in the serum and plasma of patients.

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IntroductionCorticosteroid therapy of postdiphtheric paralysis was first reported by H. J. Roberts in 1956l. We also had used corticosteroid therapy in three cases of postdiphtheric paralysis with excellent results and suggested the possible involvement of allergic factors in the etiology of this disease2.As for diphtheria, combined use of antibiotics and cortisosteroids had beell reported by several authors3, 4, 5 , 6. However, limited use of corticosteroids for the alleviation of symptoms and signs caused by diphtheria bacilli has not yet been reported.In order to determine the beneficial effect of corticosteroids on the clinical course of diphtheria, 9 patients were treated with corticosteroids alone in an initial stage which was then followed by antibiotics. Material and MethodsThe patients included the total number admitted to the Department of Pediatrics, Gunma University School of Medicine, during the period between September, 1962, and August, 1963. They consisted of 4 males and 5 females and ranged in age between 2 years and 13 years except for one 30-year-old female. I n all cases the diagnosis was clinically and bacteriologically established. Treatment before admission:Neither antibiotics nor antiserum were given before admission. Findings a t admission:Fever: Six out of 9 cases had fever and 3 were afebrile: 4 were in the 37°C range; 1 in the 38°C range; and 1 in the 39°C range.Pseudomembrane : All cases had pseudomembranes in the pharynx. Depending on size and nature, they were divided into four groups: 1) Patches in both tonsils, (+) in 4 cases; 2) patches in one tonsil and a membrane on the other tonsil, (+ + ) in 2 ; 3) membranes on both tonsils, (+ + +) in 3.Bacteriological studies : Using stained slide preparation and culture (Arakawa media), diphtheria bacilli were identified in all cases. Type differentiation and virulence tests were not made.