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Objective: To evaluate the usefulness of our original liquid-based cell preparation system AMAPS (aspiration material preparation system) and to compare it with the AutoSmear system in breast aspiration cytology. Study Design: A total of 487 specimens of fine-needle aspiration cytology of the breast were retrieved, of which 250 were processed with AMAPS and 237 with the AutoSmear method (before the introduction of AMAPS). A final histological diagnosis was obtained by an excisional biopsy or a surgical resection in 148 cases. Results: Cell recovery rates were significantly improved with AMAPS (96.8 and 99.1% in Papanicolaou and Diff-Quik, respectively) compared with the AutoSmear method (40.9 and 42.3%, respectively; p < 0.01). Within-run and day-to-day reproducibility of cell recovery was satisfactory, with coefficients of variations of 6.8 and 8.7%, respectively. Following the introduction of AMAPS in breast cytology, the unsatisfactory rate decreased significantly (from 16.0 to 8.8%; p < 0.01), while the diagnostic sensitivity for malignancy did not change (97.8 to 98.1%). Moreover, the diagnostic specificity for benign lesions increased from 75 to 93.8%, thus decreasing the excision rate of fibrocystic disease. Conclusion: AMAPS may serve as an alternative to the conventional technique or commercially available liquid-based cytology systems.

The effects of ifenprodil on respiratory activity in both the presence and absence of cyanide were examined using guinea pig brain mitochondria and mouse brain cytochrome oxidase preparations. Ifenprodil stimulated respiratory activity, exhibiting maximal effect at a concentration of 0.8 m m in the presence of succinate and potassium phosphate (Pi) among the various substrates, anions, and phosphonate compounds tested. The stimulatory action of ifenprodil on respiration could not be observed with frozen and thawed brain mitochondria. The stimulatory action was not affected by oligomycin but was reduced markedly by mersalyl. Dinitrophenol and flufenamic acid did not require the presence of Pi for their stimulatory action on respiration. Ifenprodil abolished completely the cyanide-induced inhibition of respiration with succinate as a substrate, but elicited no significant effects against the cyanide-induced inhibition of cytochrome oxidase activity in both in vivo and in vitro experiments. The results suggest that ifenprodil seems to act on the mitochondrial membrane by accelerating Pi and succinic acid transport through the membrane as well as by stimulating succinate oxidation, and that these stimulatory actions of ifenprodil may be involved in its protective effects against cyanide toxicity.

In finding that 2-(4-benzyl-piperidino)-1-(4-hydroxyphenyl)-1-propanol (BPHPP) produced high concentrations of blood glucose in rats, it was observed that (1) the maximum concentration of blood glucose was obtained 1 hr after oral administration at a dose of 100 mg/kg of BPHPP, (2) that BPHPP has no effect on the secretion of adrenal corticosterone, (3) that BPHPP markedly inhibited cAMP phosphodiesterase activity, in vitro, and (4) that in the preliminary experiment, the hepatic cAMP levels increased 1.7-fold by the administration (10 mg/kg) of BPHPP. From these results, it was presumed that the high concentrations of blood glucose may be the results of elevations in the intracellular cAMP levels due to the inhibitory action of BPHPP on cAMP phosphodiesterase.

Effects of ifenprodil (2-(4-benzyl-piperidino)-1-(4-hydroxyphenyl)-1-propanol) tartrate on the cardiovascular system were studied in mongrel dogs anesthetized with pentobarbital sodium and with alpha-chloralose and in rabbits. Ifenprodil (12.5 to 5,000 mug/kg i.v.) lowered systemic blood pressure in a dose-dependent manner in association with increased heart rate and augmented myocardial contractile force in dogs. Fall in blood pressure and tachycardia caused by ifenprodil were inhibited by pretreatment with phenoxybenzamine (5 mg/kg). The pressor responses to adrenaline and bilateral carotid occlusion were moderately reduced by pretreatment with ifenprodil (5 mg/kg), while the response to noradrenaline was attenuated only slightly. Ifenprodil (less than 1 mg/kg i.v.) decreased the total peripheral resistance and resistances of common carotid, internal carotid, vertebral, coronary, superior mesenteric, renal and femoral vascular beds in dogs. Ifenprodil increased the vertebral and femoral blood flow consistently (93.6 and 174.0%, respectively) but decreased the renal blood flow. Intra-arterial injection of ifenprodil (0.01 to 5 mug/kg) produced a dose-dependent increase in blood flow of vascular beds tested. The femoral and vertebral arterial blood flows were in particular consistently increased. Increases in the vertebral and femoral blood flows following i.v. and i.a. injections of ifenprodil were only slightly attenuated by pretreatment with phenoxybenzamine. Intravenous injections of ifenprodil (12.5 to 5,000 mug/kg) produced tachycardia in conscious rabbits but caused bradycardia after pretreatment with reserpine. From these results it was revealed that ifenprodil causes a fall in systemic blood pressure and an increase in the blood flow particularly in vertebral and femoral vasculatures following the vasodilating action directly on vascular smooth muscle. Blocking action of alpha-adrenoceptors in vessels was also demonstrated.

Summary Dialysis of potato 105,000 × g supernatant resulted in a dramatic increase in 3′, 5′- cyclic AMP phosphodiesterase activity, due to the removal of some unknown inhibitor of small molecular size, through this process. The inhibitor obtained by gel filtration was resistant to boiling at neutral, acidic, or alkaline pH and also to charcoal treatment. Kinetic analysis showed that inhibition was dependent on inhibitor concentration in the reaction mixture and that it was noncompetitive.

An inhibitor of s-glucuronidase was isolated from porcine sublingual gland by successive fractionation of trypsin extracts of the latter on Sephadex G-100, DEAE-cellulose, Sephadex G-200, and DEAE-cellulose. Its purity and homogeneity were established by DEAE-cellulose column chromatography, ultracentrifugation, and electrophoresis on cellulose-acetate membrane. The sedimentation coefficient of the purified s-glucuronidase inhibitor was 3.75 S (S200, w), and the molecular weight was determined to be 340 000 from Sephadex G-200 column chromatography. The inhibitor contained 17.5% protein, 20.8% total hexoses, 19.9% hexosamine, 21.8% N-acetylneuraminic acid, and 9.6% fucose. The inhibition was non-competitive, and it was completely suppressed by the addition of NaCl, KCl, Na2SO4, or CaCl2, respectively.

An adequate amount of typhoid-paratyphoid vaccine (TPV) or bovine liver lipid which had been emulsified in bovine serum was injected intravenously to rabbits of two groups two times at an interval of 4 days. After the second sensitization, it was found that a new specific substance had appeared in the circulating blood of each rabbit, and that it had accelerated the endogenous respiration of mouse liver cells remarkably under aerobic conditions.An effective serum was used as a starting material and examined first for stability against heat, acid, and alkali and dialyzability agains semi-dialytic cellophane membrane. Then, the extraction and purification were carried out by the column-chromatographic method. In this method, the deproteinized serum filtrate was absorbed on Amberlite IR 120H+ resin with 100-200 meshes. After that, the effective substance was extracted as a single substance through the column-chromatographic fractionation by effusion of 0.1N-HCl solution. Finally, it was identified as uric acid by paper chromatography, continuous ultraviolet and infrared absorption spectra, element dialysis, and measurement of melting point. On the other hand, uric acid was compared with some metabolic intermediates of ATP, with the exception of uric acid itself, as substrates for the oxidative respiration of mouse liver homogenate or mitochondria.

This experiment was tried to extract and purify the substance which accerelatesboth the promotion and production of antibody and to determine its molecular structureAt first, according to the method reported in the previous report, a crude effective substance was extracted by the treatments with Amberlite IR 120 H+ resin and hydrochloric acid in serum which was separated from the rabbit's blood in the early stage after the second vaccination of typoid vaccine containing O antigen.Secondly, the crude substance was repeatedly treated, taking the following procedures: i. e. it was dissolved in water, absorbed with an active animal carbon-powder and filtrated, then condensed and finally was purified into a needle crystal.Thirdly, the physical and chemical properties of the substance thus purified coincided completely with the characters of the substance which had been reported several times before. Nevertheless it did not exhibit the promoting and producing actions of antibody. From this fact, the purified substance could not be determined to be the very object. But it was determined to be “hypoxanthine” as the results of colorimetriclphenomena, protein-reactions, elementary analyses, paperchromatograms and spectro-photometric measurements by ultraviolet- and infrared-ray, etc.

L-Cystine dihydrazide and L-cystine bis (p-alkoxyanilide) derivatives listed in Tables I, II, and III were prepared and submitted to antibacterial tests with Mycobacterium tuberculosis. Two methods were used for the synthesis of these compounds. The one was du Vigneaud's method and the other was a new route to cysteinyl peptides. In the antibacterial test in vitro, the strongest activity among these compounds was found in L-cystine bis (isonicotinoylhydrazide) which was inhibitory in a concentration of 0.1γ/cc. L-Cystine bis (p-pentyloxyanilide) showed antibacterial action in the range of 1∼0.3γ/cc.

The present investigation was made to clarify the action of adrenalin on the formation of inosine, which was identified as antibody-promoting factor. The results obtained were as follows.Adrenalin displayed its action at two locations in the course of metabolic degradation of ATP and adenosine. At one location, it accelerated the degradation from ATP or adenosine to inosine and hypoxanthine, and at the other it inhibited the conversion from inosine and hypoxanthine to xanthine. Especially, when adrenalin and adenosine were used together, inosine and hypoxanthine were found to have increased about 1.7 times after incubation for 40-60 minutes, but xanthine did not behave in the same manner. The optimal rate of adrenalin was 100μg per ml of basic medium.

The possible mechanisms involved in the stimulatory effect of ifenprodil on respiration of guinea pig brain mitochondria were studied as compared with those of 2,4-dinitrophenol (DNP). Ifenprodil did not stimulate mitochondrial respiration in the absence of inorganic phosphate and the presence of oligomycin, but DNP did. The respiration with ifenprodil was not affected but that with DNP was markedly potentiated by nucleotides such as ATP, GTP, CTP, ITP, and UTP. Ifenprodil and DNP added after succinate similarly stimulated succinate oxidation, but, when added before succinate, DNP was less effective. Both ifenprodil and DNP inhibited succinate dehydrogenase activity of mitochondria. The stimulatory effect of ifenprodil on respiration with Tris-succinate as substrate was enhanced in a concentration-dependent manner by Tris-phospate at concentrations of 0–4 m m , reached plateau at 4–8 m m and declined at more than 8 m m . However, in the presence of K + , Tris-phosphate caused a concentration-dependent facilitation in the effect of ifenprodil at concentrations ranging from 0 to 40 m m ; this facilitation was far more marked than in the presence of Na + or in the presence of both Na + and K + . When Tris-succinate was substrate, Na + decreased markedly the respiratory control index (RCI) of mitochondria, but K + did not. Mg 2+ also decreased the RCI of mitochondria and reduced the stimulatory effect of ifenprodil on respiration. These findings suggest that ifenprodil differs from DNP in a mode of stimulatory action on mitochondrial respiration and stimulates respiration by presumably affecting intramitochondrial levels of K + , phosphate, and succinate.

In in vivo experiments, dose-dependent protection against the lethality of cyanide was elicited by im pretreatment with ifenprodil in rats, guinea pigs, rabbits, and mice. The decline in respiration rate, blood pressure, and heart rate, and the flat electroencephalogram induced by iv administration of sodium cyanide was also reversed by previous administration of ifenprodil in these species. Methemoglobinemia was not observed after protective doses of ifenprodil in rats and ifenprodil did not affect thiocyanate formation induced by sodium cyanide and sodium thiosulfate in rats. In vitro, ifenprodil exhibited a stimulative effect on respiration of mitochondria isolated from guinea-pig liver, this effect being the most prominent with succinate as substrate and in the presence of phosphate. In addition, the inhibitory effect of potassium cyanide on the respiration was completely abolished by ifenprodil. These protective effects of ifenprodil on cyanide intoxication were also observed with both ifenprodil tartrate and hydrochloride, and tartaric acid did not exert this action. Experiments using ultraviolet absorption and proton nuclear magnetic resonance spectra did not show that ifenprodil reacts directly with free cyanide ion to trap it. The results suggest that ifenprodil prevents the signs and lethality of cyanide, and a part of the mechanism can be ascribed to antagonism at the mitochondrial level, though the mechanism is not clear at present.

For the purification of the bacterial thiaminase II produced by Bacillus aneurinolyticus Kimura et Aoyama, the 7-day culture in the synthetic medium was subjected to osmotic concentration using the equipment devised by the authors and fractional saturation with ammonium sulfate. The crude enzyme obtained with 0.3-0.65 ammonium sulfate saturation was passed through each of a column of cation or anion exchange resin or through a column of both kinds of resin. By these procedures the enzyme activity was highly increased. The addition of chelate reagents, particularly EDTA, was proved to be accerelative and a crystalline substance presumably of the thiaminase could be obtained. In the process of purification the enzyme activity and absorption spectrum of each fraction were determined. Some properties of the purified preparation were reported.