The essential criteria for a useful analytical technique are specificity, sensitivity, accuracy, precision, simplicity, rapidity and economy. The sensitivity and specificity of immunoassays have been improved by utilizing sensitive marker, monoclonal or clonotypic antibodies and adequate conditions for antigen-antibody reaction. Applications of various immunoassays to laboratory medicine make it possible to elucidate etiology of various diseases and make early diagnosis and prevention of the diseases such as congenital hypothyroidism and subclinical prolactinoma. Since the immunoassay has many advantages, its application should result in remarkable progress in laboratory medicine.
Tsutsugamushi disease and its etiological agent, Rickettsia tsutsugamushi, were discovered by Japanese pioneers, followed by its appearance not only almost everywhere Japan, but also in China and Southeast Asia. The incidence of the disease in our country decreased after the Second World War and, between 1965 and 1975, only a few patients were detected. However, since 1976, the number of patients with this disease in Japan started to increase and, recently about 900 patients were dectected every year. In the initial part of this review, the author refers to the recent profile of Tsutsugamushi disease in Japan, and recent knowledges about the relation of vector and rickettsia, and in the latter part, reviews the progressing studies on R. tsutsugamushi obtained in the author's laboratory, such as, the development of experimental technologies, rickettsial multiplication cycle in the host cell, discovery of various antigenic variants, chemical and antigenic structures of the rickettsia, and finally, mentions about the future problems on the disease and the rickettsia.
Paterno-Buchi reaction of benzaldehyde and 2, 3-dihydrofuran derivatives was studied. 2, 3-Dihydrofuran formed two stereoisomers of 7-phenyl-2, 6-dioxabicyclo [3. 2. 0] heptanes (Y type) selectively. Other 2, 3-dihydrofuran derivatives induced by acid catalyzed cleavage of 6-ethyl-2, 7-dioxabicyclo [3. 2. 0] hept-3-ene gave 2, 6-dioxabicyclo [3. 2. 0] heptane rings (Y type addition) and 2, 7-bicyclo [3. 2. 0] heptane rings (X type addition) in a special case. Photocycloaddition of 6-ethyl-2, 7-dioxabicyclo [3.2.0] hept-3-ene (Ib) formed two stereoisomers of 3, 6, 8-trioxatricyclo [5.2.0.02, 5] nonane skeltons (Y type) and two 4, 6, 8-trioxatricyclo-[5.2.0.02, 5] nonanes (X type). The formation of X type oxetanes was observed when an intermediate biradical was stabilized by a resonance effect. Stereoselectivity was established at the formation of X type oxetanes to form threo-aldol equivalents.
From the fronds of Costarican fern Odontosoria gymnogrammoides CHRIST (Pteridaceae), a phenolic glycoside 'odontoside (I)', was isolated together with protocatechuic acid, protocatechualdehyde, orientin (II) and 6"-O-acetyl orientin (III, a new compound). The structure of I was unequivocally established by comparing 13C nuclear magnetic resonance spectra of I and 3, 4-dimethoxybenzyl dimethylprotocatechuate, which was synthesized by the macrolidelactonization method using cesium carbonate from dimethylprotocatechuic acid and 3, 4-dimethoxybenzyl alcohol.
From Ophiopogon tuber (China), a new borneol derivative (III) and a new steroidal sapogenin named ophiogenin (IV) were isolated in addition to glycerol (I) and α-humulene (II). The structures of III and IV were established as calcium bornyl sulfate and 25R-spirost-5-en-3β, 14α, 17α-triol on the basis of several spectral data. The antimicrobial activities for the isolated constituents from Ophiopogon tuber were examined. As a result, 25S-ruscogenin and a mixture of ruscogenin and 25S-ruscogenin showed antimicrobial activities on Gram-positive bacteria.
Our previous work has indicated that there is no significant difference between the rates of glycogen synthesis from the α and β anomers of glucose in the rat diaphragm. The present study was performed to prove this fact by means of mathematical estimation of the glycogen synthesis rate. In order to obtain the values of parameters necessary for the mathematical estimation, we measured the rate of glucose uptake by the rat diaphragm, the contents of glucose and glucose-6-phosphate in the tissue, and their mutarotational rates. As a result of calculation, the rate of glycogen synthesis from α-D-glucose was shown to be indistinguishably similar to that from β-D-glucose, supporting our previous data. During the course of this study, the presence of compartmentation of glucose metabolism in the rat diaphragm was also suggested.
Sodium bisulfite is often employed as an antioxidant in injections. However, residual bisulfite in some commercial preparations was largely responsible for the cleavage of fulsultiamine and thiamine in parenteral admixtures during storage. Results are presented on the rate of fulsultiamine cleavage by variation of the bisulfite concentration (0-4×10-4M). Plots of the logarithm of percentage of residual fulsultiamine against time were linear and the cleavage was first-order with respect to fulsultiamine. It was also found that fulsultiamine and thiamine were stabilized by glucose in the admixture ; i.e., without the addition of glucose the degradation rate constant was 738M-1h-1 for fulsultiamine and 33.8M-1h-1for thiamine in aqueous solution of pH 5.0 at 20°C, and by the addition of 50% glucose the rate constant decreased to 95M-1h-1 for fulsultiamine and 4.5M-1h-1 for thiamine, respectively.
Complexes of p-nitrophenol and p-nitrophenolate anion with heptakis-(2, 6-di-O-methyl)-β-cyclodextrin (DM-β-CD) in aqueous and organic solutions have been studied by spectroscopic analyses. Inclusion complex formation was observed on the basis of the changes of ultraviolet-visible absorption spectra in aqueous and organic solutions. The stability constants of the complexes were determined by the Benesi-Hildebrand equation. It was found that the stability constant of p-nitrophenolate anion-DM-β-CD complex was greater than that of p-nitrophenol-DM-β-CD complex in the aqueous solutions. Proton nuclear magnetic resonance data showed that the mode of complex formation of p-nitrophenol-DM-β-CD was the same as that of p-nitrophenolate anion-DM-β-CD in D2O. The apparent pKa value of p-nitrophenol decreased with increasing concentration of DM-β-CD.
The kinetic dissolution behaviour of carbamazepine (CBZ) polymorphs, three anhydrous forms (I, II, and III forms) and one dihydrate form (IV form), were investigated in detail by the rotating disk and dispersion methods. All of anhydrous forms showed a simultaneous phase transition from an anhydrous form to a dihydrate form during dissolution. Accordingly, a model involving a term of phase transition during dissolution was used, and the solubilities of anhydrous forms, Caa, and the rate constants of crystallization process, kr, were calculated by the rotating disk method. I and II forms showed the highest values of the solubility and the phase transition rate, respectively. Furthermore, the values of Caa and kr obtained by the dispersion method used in the above model adding a term of variable particle size were in fair agreement with those by the rotating disk method.
Metabolism of 6-[(R)-2-[3-(3, 4-dihydroxybenzoyl)-3-(3-hydroxypropyl)-1-ureido]-2-phenylacetamido] penicillanic acid (AO-1100), a new ureidopenicillin having catechol moiety, was studied in vitro, using catechol-O-methyltransferase (COMT) prepared from the rat and rabbit livers. Main two metabolites were detected by means of high performance liquid chromatography and identified as the monomethyl derivatives (M1, M2) of AO-1110 by the total synthesis. It was found that AO-1100 was metabolized easily by COMT prepared from the rat liver, and not easily by COMT from the rabbit liver. The metabolites (M1, M2) showed greatly reduced antibacterial activity against gram-negative bacteria including Pseudomonas aeruginosa.
In order to examine antifungal activity, several amino derivatives of 2-phenyl-3 (2H)-pyridazinone were synthesized. Among the compounds tested, 2-(p-aminophenyl)-4, 5, 6-trichloro-3 (2H)-pyridazinone (IId) and 4-amino-6-chloro-2-(p-nitrophenyl)-3(2H)-pyridazine-thione (IX) revealed strong antifungal activities in vitro.
In the course of screening of natural products for Ca2+-blocking activity in the rat thoracic aorta, the extract of Boenninghausenia albiflora var. japonica was found to have such activity. The extract was further fractionated by column chromatography to identify the active constituents. The active principle was identified as a coumarin derivative, chalepensin. In addition, the inhibitory effects of coumarin related compounds, such as coumarin, umbelliferone, 7-isopentenyloxycoumarin, aurapten, esculin, esculetin, esculetin 6, 7-dimethyl ether, decursin, psoralen, bergapten, xanthotoxin and imperatorin were examined. Decursin and imperatorin showed inhibitory effects similarly to that of chalepensin and two orders of magnitude less active than verapamil.
Five flavones dioxygenated at C-2'and 6'(5, 7, 2', 6'-tetrahydroxy-, 5, 7, 2'-trihydroxy-6'-methoxy-, 5, 2'-dihydroxy-7, 8, 6'-trimethoxy-, 5, 2', 6'-trihydroxy-7, 8-dimethoxy-, and 5, 7, 2'-trihydroxy-8, 6'-dimethoxyflavone) isolated from Scutellaria spp. were synthesized to confirm their structures. Spectroscopic comparisons of the synthetics with the corresponding natural flavones confirmed the respective structures.
Effects of hydrolizate (III·HCl) of Flutazolam (I) on the antianxiety, the anticonvulsion, the prolongation of the sleep, the muscle relaxation and the antihypoxia were compared with I in mice by the established methods. It was demonstrated that III·HCl inhibited the anxiety and pentetrazol-induced convulsion, prolonged the thiopental-induced sleep, and produced the skeletal muscle relaxation and the protection against hypobaric-hypoxia. These potencies of III·HCl were equipotent to I. These results suggested that III·HCl might be able to be delivered in an injection form.