Adenosinetriphosphate (ATP) synthase (FoF1) is a major energy supplying enzyme of cells utilizing the proton motive force. It consists of a catalytic portion called F1 and a proton channel portion called Fo. In order to elucidate the chemical reaction of FoF1, thermophilic FoF1 (TFoF1) was used, because it is stable and could eb reconstituted without Mg-ATP. In contrast to the previous hypotheses on the ATP synthesis, direct measurement of H+ current through TFoF1 incorporated into a planar lipid bilayer, 3H+/ATP stoichiometry was obtained. The primary structure of TFoF1 was established by sequencing its operon deoxyribonucleic acid and subunit peptides. The stereochemistry of the reaction using [16O, 17O, 15O, 35S] thiophosphate supported the a pathway for associative nucleophilic displacement on a phosphoric ester without pseudorotation. The diastereoisomeric preference of Cd-ATPγS revealed that the true substrate of TFoF1 is Δ, β, γ, bidentate Mg-ATP, like adenylate kinase. The site directed mutagenesis of the residues of F1 homologous to Mg-ATP binding site of adenylate kinase revealed their essential role in the reaction. Mitchell's chemiosmotic theory was refined by these results.
Knowledge which will influence someone upon his decision making is "information"for him. Physicians have to make their decision when they prescribe medicines or give orders for injections. Recently highlevel activities for accurate selection of information are indispensable, as a result of increases in the volume and complications of the quality of drug-related information. The drug information provided by hospital pharmacists for the past 20 years has been successful, but does not always satisfy physicians, according to the substantial difference in approaches to drug information between physicians and pharmacists. I considered if a computer will be able to fill the gap. In 1980 we developed and reported in the field of medical informatics, a system for automatically checking prescriptions, an automatic dispensing apparatus, and a series of system for the administration of consumption and stock of drugs, and their automatic order execution. We have further continued to investigate database architecture to incorporate all pharmaceuticals in the NIH Price List (ca. 16 000), the data being classified into 33 items and retrievable from 14 items. I would like to introduce another plan for automatic data supply service being developed in our center. The full texts have been input of package inserts for all pharmaceuticals currently on the japanese marker (ca. 30 000) using the TOSfile 3200 optical disk system, so that comprehensive drug information is available on a 24-h basis to medical or pharmaceutical professionals throughout Japan by facsimile transmission.
3-Arylamino-2-dialkylaminomethyl-4, 4-dimethyl-2-buten-4-olides and their aza analogs were synthesized from 5, 5-dimethyltetronic acid and tetramic acid, and tested analgesic and antimicrobial activities.
A new biflavonoid, daphnodorin D (5) was isolated from the root of Daphne odora THUNB. Daphnodorin D was confirmed to be a mixture of atropisomers at C-8, C-3"linkage of 5, 7, 4'-trihydroxyflavan〓 5", 7", 4'''-trihydroxyflavone by chemical and spectroscopic means.
The insulin-like effects and the promotions of insulin release of hot-water extracts from various kinds of flesh, including meat, chicken, fishes and shellfish were studied in vitro. An examination of insulin-like effect, in which the indicator was the conversion of D-[2-3H]-glucose to lipids in free fat cells of rats, indicated the existence of insulin-like effects in flesh such as Spanish mackerel (Scomberomorus niphonius) and bonito (Katsuwonus pelamis) of Scombridae, in shellfish like abalone (Haliotis gigantea) and shortnecked calm (Tapes philippinarum), and in the flesh and fowl such as pig (Sus scrofa) and chicken (Gallus domesticus). An examination of insulin release promotion effect, in which the indicator was the release of insulin from the isolated Langerhans'islands of the hamster, indicated that flesh, fowl, fishes and shellfish used in this experiment exhibited an effect which promotes the release of insulin.
New hydrogen donors, water soluble lophine derivatives, were synthesized for the determination of hydrogen peroxide developed in the enzymatic reaction of biological substances. These chromogens have high absorptivity ; extinction coefficient is 40 000-50 000 upon oxidation with hydrogen peroxide in the presence of peroxidase. A single work reagent is used for the determination of uric acid in the serum ; the reaction is complete in less than 5 min at 37°C. The blue dye is formed at 640 nm ; a blank sample measurement is not needed. The standard curve for the method is linear for uric acid ranging from 2.5 to 25 mg/dl. Average analytical recovery of uric acid in the human sera exceeded 99% ; within-run and between-run precision studies showed CV's ⪈2.5 and ⪈3.5%, respectively. The method correlated well with the standard uricase-ultraviolet method ; the regression equation is y=0.988x+0.426 (mg/dl) and the correlation coefficient (γ) is 0.977 (n=11).
Escolar (Lepidocybium flavobrunneum) belonging to family Gempylidae are caught by a rope stretching fishing. The sale of escolars for food, however, is prohibited in Japan because their muscles are rich in wax which may cause a food poisoning. In order to use this wax as a base of medicine or cosmetics, the purification and refinement of the wax and its safety were investigated. Escolar's muscle contained about 20% lipid and its 88.8% consisted of wax. The wax was composed of C32, C34, C36 and C38 and the main component was C34H66O2. The components of alcohols, as unsaponifiable substances of escolar wax, were mainly C16 : 0 and C18 : 1, as well as those of sperm whale (Physeter catodon) wax. The components of fatty acids, as saponifiable substances of the wax, was mainly C18 : 1 and those of highly unsaturated fatty acids a little. Escolar wax was refined by deacidification, decolorization with activated white soil, hydrogenation with Cu-Cr catalyst and molecular distillation. The refined wax was semisolid at room temperature and smelled faintly like margarine. Its safety was asseyed by acute toxicity test, skin irritation test, eye's mucous membrane irritation test and contact sensitization test. The results showed that the refined wax was highly safe, indicating the usefulness for a base of medicine of cosmetics.
Extra weak chemiluminescence (CL) from 30 kinds of crude drugs observed during extraction was measured. Among these drugs, Atractylodis Lanceae Rhizoma, Zingiberis Rhizoma, Ginseng Radix, Magnoliae Cortex, Astragali Radix, Rehmanniae Radix, Cnidii Rhizoma, Glycyrrhizae Radix, Bupleuri Radix and Fritillariae Bulbus showed high CL intensities. Furthermore, the emission spectra of these drugs resembled each other and had emission bands identical with those from active oxygen. CL generated from Glycyrrhizae Radix was easily quenched by replacing air or oxygen gas by nitrogen gas. It was also quenched effectively in the presence of antioxidants such as ascorbic acid or thioglycerol. The water extract of Glycyrrhizae Radix was fractionated by column chromatography. The fractions containing triterpenoids, flavones, cumarins, amino acids and amino acid derivatives generated intense CL. However, the authentic solution of glycyrrhetic acid, liquiritin or amino acids did not show CL. It was assumed that CL produced on extraction of Glycyrrhizae Radix originated from active oxygen generated by oxidation of amino acid derivatives. This chemiluminescence technique may be useful as a monitor of the quality of Kampo Herbal Drugs.
The mechanism of interaction of furosemide (FS) with chloral hydrate (CH) was investigated in rabbits. The co-administration of CH markedly enhanced the plasma concentration of FS, and reduced the amount of cumulative urinary excretion of FS after intravenous bolus injection. Furthermore, the co-administration of CH was found to significantly decrease the renal clearance (Clr) of FS, but to have no effect on its extrarenal clearance (Clnr). These clearly indicate that the interaction of FS with CH is generated during renal excretion. Interestingly, the co-administration of CH reduced the amount of cumulative urinary excretion of phenolsulfonphthalein, which is excreted exclusively by tubular secretion, whereas it did not affect the amount of cumulative urinary excretion of inulin, which is excreted exclusively by glomerular filtration. The co-administration of CH caused no change in urinary pH after intravenous bolus injection of FS. These findings suggest that the co-administration of CH may depress the renal excretion of FS by inhibiting the tubular secretion.
The oxidized metabolites appeared in the rat plasma after intravenous administration of benzenesulfonamide (Ia), 1-cyclohexyl-3-benzenesulfonylurea (Ib) and 1-n-butyl-3-benzene-sulfonylurea (Ic) with a (±)-α-hydroxyethyl substituent at the para position were investigated. The metabolically produced ketones were identified with those of authentic samples by comparing their high performance liquid chromatographic retention times. On the other hand, Ib (hydroxyhexamide) and its major metabolite, IIb (acetohexamide) extracted from the rat plasma, were identified by gas chromatography-mass spectrometry after trimethyl-silylation. These alcohols and respective ketones are presumably related to reversible drug-metabolite in rats.
In order to examine analgesic activity, 6-alkoxy-4-N, N-(dialkylamino) (or morphorino)-alkylamino-2-phenyl-3 (2H)-pyridazinones (III) were synthesized. Among the compounds tested, 4-(3-dimethylaminopropylamino)-6-ethoxy-2-phenyl-3 (2H)-pyridazinone (IIIg) revealed about six-fold strong activity of that of aminopyrine in an analgesic test by the modified Haffner's method. The safety margin of IIIg as an analgesic agent was considerably higher than that of aminopyrine.
From the leaves of Artocarpus integra MERR., which is used as a herbal drug in Indonesia, (±)-catechin was isolated as an active principle with an inhibitory effect on stress ulcer formation in restrained and water-immersed mice. The structure of (±)-catechin was elucidated on the basis of spectral comparison with authentic (+)-catechin.
D-Cysteinolic acid (2-L-amino-3-hydroxy-1-propane sulfonic acid) isolated from sardine Sardinops melanostica was tested for pharmacological activities along with taurine. It inhibited the aggregation of rat platelet induced by collagen, thrombin and ADP, more effectively than taurine. However, it was less effective than taurine on the stability of rat erythrocyte membrane and inhibitory activity of gastric juice secretion in pylorus-ligated rat, and not effective on the blood pressure of carotid artery in spontaneously hypertensive rat.