• The Korean Journal of Pharmacology
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• v.16 no.2 s.27
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• pp.55-70
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• 1980

The pharmacological and microbiological studies of Cefoperazone (T-1551, Toyama Chemical Co., Japan) were conducted in vitro and in vivo. The studies included stability and physicochemical characteristics, antimicrobial activity, animal and human pharmacokinetics, animal pharmacodynamics and safety evaluation of Cefoperazone sodium for injection. 1) Stability and physicochemical characteristics. Sodium salt of cefoperazone for injection had a general appearance of white crystalline powder which contained 0.5% water, and of which melting point was $187.2^{\circ}C$. The pH's of 10% and 25% aqueous solutions were 5.03 ana 5.16 at $25^{\circ}C$. The preparations of cefoperazone did not contain any pyrogenic substances and did not liberate histamine in cats. The drug was highly compatible with common infusion solutions including 5% Dextrose solution and no significant potency decrease was observed in 5 hours after mixing. Powdered cefoperazone sodium contained in hermetically sealed and ligt-shielded container was highly stable at $4^circ}C{\sim}37^{\circ}C$ for 12 weeks. When stored at $4^{\circ}C$ the potency was retained almost completely for up to one year. 2) Antimicrobial activity against clinical isolates. Among the 230 clinical isolates included, Salmonella typhi was the most susceptible to cefoperazone, with 100% inhibition at MIC of ${\leq}0.5{\mu}g/ml$. Cefoperazone was also highly active against Streptococcus pyogenes(group A), Kletsiella pneumoniae, Staphylococcus aureus and Shigella flexneri, with 100% inhibition at $16{\mu}g/ml$ or less. More than 80% of Escherichia coli, Enterobacter aerogenes and Salmonella paratyphi was inhibited at ${\leq}16{\mu}/ml$, while Enterobacter cloaceae, Serratia marcescens and Pseudomonas aerogenosa were somewhat less sensitive to cefoperagone, with inhibitions of 60%, 55% and 35% respectively at the same MIC. 3) Animal pharmacokinetics Serum concentration, organ distritution and excretion of cefoperazone in rats were observed after single intramuscular injections at doses of 20 mg/kg and 50 mg/kg. The extent of protein binding to human plasma protein was also measured in vitro br equilibrium dialysis method. The mean Peak serum concentrations of $7.4{\mu}g/ml$ and $16.4{\mu}/ml$ were obtained at 30 min. after administration of cefoperazone at doses of 20 mg/kg and 50 mg/kg respectively. The tissue concentrations of cefoperazone measured at 30 and 60 min. were highest in kidney. And the concentrations of the drug in kidney, liver and small intestine were much higher than in blood. Urinary and fecal excretion over 24 hours after injetcion ranged form 12.5% to 15.0% in urine and from 19.6% to 25.0% in feces, indicating that the gastrointestinal system is more important than renal system for the excretion of cefoperazone. The extent of binding to human plasma protein measured by equilibrium dialysis was $76.3%{\sim}76.9%$, which was somewhat lower than the others utilizing centrifugal ultrafiltration method. 4) Animal pharmacodynamics Central nervous system : Effects of cefoperazone on the spontaneous movement and general behavioral patterns of rats, the pentobarbital sleeping time in mice and the body temperature in rabbits were observed. Single intraperitoneal injections at doses of $500{\sim}2,000mg/kg$ in rats did not affect the spontaneous movement ana the general behavioral patterns of the animal. Doses of $125{\sim}500mg/kg$ of cefoperazone injected intraperitonealy in mice neither increased nor decreased the pentobarbital-induced sleeping time. In rabbits the normal body temperature was maintained following the single intravenous injections of $125{\sim}2,000mg/kg$ dose. Respiratory and circulatory system: Respiration rate, blood pressure, heart rate and ECG of anesthetized rabbits were monitored for 3 hours following single intravenous injections of cefoperazone at doses of $125{\sim}2,000mg/kg$. The respiration rate decreased by $3{\sim}l7%$ at all the doses of cefoperazone administered. Blood pressure did not show any changes but slight decrease from 130/113 to 125/107 by the highest dose(2,000 mg/kg) injected in this experiment. The dosages of 1,000 and 2,000 mg/kg seemed to slightly decrease the heart rate, but it was not significantly different from the normal control. All the doses of cefoperazone injected were not associated with any abnormal changes in ECG findings throughout the monitering period. Autonomic nervous system and smooth muscle: Effects of cefoperazone on the automatic movement of rabbit isolated small intestine, large intestine, stomach and uterus were observed in vitro. The autonomic movement and tonus of intestinal smooth muscle increased at dose of $40{\mu}g/ml$ in small intestine and at 0.4 mg/ml in large intestine. However, in stomach and uterine smooth muscle the autonomic movement was slightly increased by the much higher doses of 5-10 mg/ml. Blood: In vitro osmotic fragility of rabbit RBC suspension was not affected by cefoperazone of $1{\sim}10mg/ml$. Doses of 7.5 and 10 mg/ml were associated with 11.8% and 15.3% prolongation of whole blood coagulation time. Liver and kidney function: When measured at 3 hours after single intravenous injections of cefoperaonze in rabbits, the values of serum GOT, GPT, Bilirubin, TTT, BUN and creatine were not significantly different from the normal control. 5) Safety evaluation Acute toxicity: The acute toxicity of cefoperazone was studied following intraperitoneal and intravenous injections to mice(A strain, 4 week old) and rats(Sprague-Dawler, 6 week old). The LD_(50)'s of intraperitonealy injected cefoperazone were 9.7g/kg in male mice, 9.6g/kg in female mice and over 15g/kg in both male and female rats. And when administered intravenously in rats, LD_(50)'s were 5.1g/kg in male and 5.0g/kg in female. Administrations of the high doses of the drug were associated with slight inhibition of spontaneous movement and convulsion. Atdominal transudate and intestinal hyperemia were observed in animals administered intraperitonealy. In rats receiving high doses of the drug intravenously rhinorrhea and pulmonary congestion and edema were also observed. Renal proximal tubular epithelial degeneration was found in animals dosing in high concentrations of cefoperazone. Subacute toxicity: Rats(Sprague-Dawley, 6 week old) dosing 0.5, 1.0 and 2.0 g/kg/day of cefoperazone intraperitonealy were observed for one month and sacrificed at 24 hours after the last dose. In animals with a high dose, slight inhibition of spontaneous movement was observed during the experimental period. Soft stool or diarrhea appeared at first or second week of the administration in rats receiving 2.0g/kg. Daily food consumption and weekly weight gain were similar to control during the administration. Urinalysis, blood chemistry and hematology after one month administration were not different from control either. Cecal enlargement, which is an expected effect of broad spectrum antibiotic altering the normal intestinal microbial flora, was observed. Intestinal or peritoneal congestion and peritonitis were found. These findings seemed to be attributed to the local irritation following prolonged intraperitoneal injections of hypertonic and acidic cefoperazone solution. Among the histopathologic findings renal proximal tubular epithelial degeneration was characteristic in rats receiving 1 and 2g/kg/day, which were 10 and 20 times higher than the maximal clinical dose (100 mg/kg) of the drug. 6) Human pharmacokinetics Serum concentrations and urinary excretion were determined following a single intravenous injection of 1g cefoperazone in eight healthy, male volunteers. Mean serum concentrations of 89.3, 61.3, 26.6, 12.3, 2.3, and $1.8{\mu}g/ml$ occured at 1,2,4,6,8 and 12 hours after injection respectively, and the biological half-life was 108 minutes. Urinary excretion over 24 hours after injection was up to 43.5% of administered dose.

• Applied Biological Chemistry
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• v.14 no.1
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• pp.59-97
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• 1971

As a study on the cellulase of Myriococcum albomyces the culture media for enzyme formation and properties of its crude preparation were investigated and the crude enzyme preparation was further fractionated. The results are summarized as follows: 1. Wheat bran solid culture produced stronger activities of cellulase than rice bran or defatted soy bean meal solid culture. 2. Shaking culture with wheat bran, rice bran or defatted soy bean meal produced higher cellulase activities than solid culture with the corresponding media. 3. The enzyme formation was higher at $45^{\circ}C$ than at $37^{\circ}C$ or $50^{\circ}C$ regardless of the kind of culture medium. 4. The formation of CMCase activity was more promoted by organic nitrogen source than inorganic nitrogen source. 5. The formation of cellulase activities were increased 1.5 to 3.0-fold by adding CMC, Avicel or cellulose powder as an inducer into 5% wheat bran basal medium. 6. Cellulase production using a tank culture procedure with addition of CMC or Avicel as an inducer was the highest at fifth day and thereafter decreased slightly. 7. The crude enzyme preparation showed pH optimum in 4.0 to 4.5, and pH stability in the range of 3.5 to 8.0. Optimum temperature for the activity was $65^{\circ}C$ which was higher than among other cellulases and it was stable at $60^{\circ}C$ for 120 minutes. 8. Dialyzed crude enzyme was activated by $Ca^{++}$ and $Mg^{++}$, but inhibited by $Hg^{++}$, $Cu^{++}$ and $Ag^{+}$. 9. Four different types of cellulase, i. e., fraction I, fraction II-a, fraction II-b, and fraction III were purified from the culture filtrate of Myriococcum albomyces through a sequence of ammonium sulfate fractionation, and elution chromatography on DEAE-Sephadex A-25, Amberlite CG-25 type 2 and hydroxyapatite columns. 10. These four cellulase fractions were showed to be homogenous by electrophoresis and ultracentrifugation and also gave a typical ultraviolet absorption spectrum of protein. 11. Four purified fraction showed different specificity toward substrates, fraction I has a stronger activity toward Avicel, cellulose powder, and gauze than that of other cellulase fractions. Fraction II-a had a powerful activity toward cellobiose but it was almost inactive agaisnt fibrous cellulose contrary to fraction I. On the contrary, the main component fraction II-b had a fairly higher activity on CMC and Avicel. Activity of fraction II-b toward cellobiose was about one-third of that of fraction II-a and activity on Avicel was lower than that of fraction I. Fraction III had a more powerful activity in decreasing viscosity of CMC. 12. Final hydrolysis products of fibrous cellulose by each fraction were cellobiose and glucose. Whereas oligosaccharides were predominant in the early stage of hydrolysis, prolonged reaction produced more glucose than cellobiose. Fraction I and fraction II-a acted synergically on Avicel. 13. Optimum pH for the activities of cellulase fraction I, fraction II-a, fraction II-b and fraction III were found to be 5.5, 5.0, 4.0 and $4.0{\sim}4.5$, respectively. These fractions were found to be stable in the range of pH $3.0{\sim}7.5$. 14. Optimum temperature for the activities of fraction I, fraction II-a, fraction II-b, and fraction III were $50^{\circ}C$, $55^{\circ}C$, $60^{\circ}C$ and $55^{\circ}C$, respectively. No less of activity was found by heating 120 minutes at $55^{\circ}C$ and fraction II-a was more stable than the others at $60^{\circ}C$. 15. Fraction I and fraction II-b were activated by $Ca^{++}$ and $Mg^{++}$ but inhibited by $Hg^{++}$ and $Ag^{+}$.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.3
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• pp.49-82
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• 1965

For the stable and high yields of low-land rice in Korea, the characteristics of rice plant for the vegetative and physiological responses, plant type formation, and yield components have been studied in order to obtain the fundamental data for the improvement of cultural practices, especially for the ideal fertilizer application. Furthermore the environmental conditions in Korea including temperatures, light, precipitation, and soil conditions have been compared in the broad sense with those in Japan, and the application of nitrogen, phosphorus, potassium, silicate and other micro-nutrients were described in relation to the characteristics of environmental conditions for the improvement of fertilizer application. 1. The average yield of polished-rice per 10 are in Korea is about 204 kg and this values are much less than those in Japan and Taiwan where they produce 77% to 13% more than in Korea. The rate of yield increase a year in Korea is 4.2 kg, but in Japan and Taiwan the rates of yield increase a year are 81 % and 62%, respectively. It was also found that the coefficient of variation of yield is 7.7% in Korea, 6.7% in Japan and 2.5% in Taiwan. This means that the stability of producing rice in Korea is very low when compared with those in Japan and Taiwan. 2. It was learned from the results obtained from the 'annual yield estimation experiment' that there are big differences in the respect of plant type formations between rice crops grown in Japan and Korea. The important differences found were as follows: (1) The numbers of spikelets per 3.3 square meters are 891 in Korea and 1, 007 in Japan(13% more than in Korea). (2) The numbers of tillers per 3.3 square meters at the stage of maximum tillering are 1, 150 in Korea, but in Japan they showed 19% more than in Korea. (3) The ratio of effective tillers to total tillers is 77.5% in Korea and 74.7% in Japan, which seems to be higher in Korea than in Japan. But the ratio in Korea is very low when considered the numbers of total tillers in both countries. (4) The ratio of grain to straw is 85.4% in Korea and 96.3% in Japan. 3. The average temperatures during the growing season at the area of Suwon, Kwangjoo and Taegu are almost same as those in the district of Jookokoo(Fookoo yama) in Japan, i.e., the temperatures during the rice-growing season in Korea are similar to those in the southern-warm regions of Japan. 4. Considering the minimum temperatures at the stage of limiting transplanting, 13$^{\circ}C$, the time of transplanting might be 30 to 40 days earlier than presently practicing transplanting time, which comes around June 10. 5. The temperatures during the vegetative growth in Korea were higher than those temperatures that needed in the protein synthesis which ate the main metabolism during this stage. However, the temperatures at the time of reproductive growth was lower than the temperatures that needed in the sugar assimilation which is main metabolism in this stage. In this point of view, it might be considered that the proper time of growing rice plant in Korea would be rather earlier. 6. The temperatures and the day light conditions at the time of first tillering stage of rice plant, when planted as presenting transplanting practices, are very satisfactory, but the poor day light length, high temperatures and too wet conditions in the time of last-tillering stage(mid or last July) might cause the occurrence of disease such as blast. 7. The heading stage of rice plants at each region through nations when planted as presently practicing method comes when the day light length is short. 8. It was shown that the accumulated average air-temperature at the time of maturing stage was not enough and the heading time was too late, when considered the annual deviations of mean temperatures and low minimum temperatures. 9. The nitrogen content of each plant part at the each growing stage was very high at the stage of vegetative growth when compared with the nitrogen content at the stage of reproductive growth after heading. In this respect it was believed to be important to prevent the nutrient shortages at the reproductive stages, especially after the heading. 10. The area of unsatisfactory irrigation paddy fields and natural rain-fed paddy fields are getting reduced in Korea. The correlation between the rate of reducing unsatisfactory irrigation and natural rain-fed paddy fields and the rate of yield increase were computed. The correlation coefficients(r) between the area of unsatisfactory irrigation paddy fields and yield increase were +0.525, and between the natural rain-fed paddy fields and yield increase, +0.832 and between the unsatisfactory irrigation plus natural rain-fed paddy fields and yield increase, +0.84. And there were. highly significant positive correlations between natural rain-fed paddy fields and yield increases indicating that the less the area of natural rain-fed paddy fields, the greater the yields per unit area. 11. The results obtained from the fertilizer experiments (yield performance trials) conducted in both Korea and Japan showed that the yield of non-fertilized plots per 10 are was 231 kg in Korea and 360 kg in Japan. On the basis of this it might be concluded that the fertility of soil in Korea is lower than that in Japan. Furthermore it was. also found that the yields of non-nitrogen applied plots per 10 are were 236 kg in Korea and 383 kg in Japan. This also indicates that the yields of rice in Korea are largely depending on the nitrogen content in the soil. 12. The followings were obtained when the chemical natures of soils in both Korea and Japan were compared. (1) The content of organic matter, total nitrogen, exchangeable calcium, and magnesium in Korea were no more than the half those in Japan. (2) The content of N/2 chloride and soluble silicate in low-land soil were on the average lower in Korea. (3) The exchange capacity of bases in Korea was no more than half that in Japan. 13. It was also observed by comparing the soil nature of the soil with high yielding capacity with the soil with low yielding capacity that the exchange capacity of bases, exchangeable calcium and magnesium, potassium, phosphorus, manganese, silicate and iron were low in the soil with low yielding capacity. 14. The depth of furrow slice was always deeper in the soil with high yielding capacity, and the depth of furrow slice in Korea was also shallower than that in Japan. 15. Summarizing the various conditions mentioned previously and considering the effects of silicate and trace elements such as manganese and iron besides three elements on the physiological and plant type formation of rice crops, more realistic and more ideal fertilizing practices were proposed. proposed.