Keiko Yamaoka

Involvement of fibroblast growth factor 18 in dedifferentiation of cultured human chondrocytes

Objective:  Chondrocytes inevitably decrease production of cartilaginous matrices during long‐term cultures with repeated passaging; this is termed dedifferentiation. To learn more concerning prevention of dedifferentiation, we have focused here on the fibroblast growth factor (FGF) family that influences chondrocyte proliferation or differentiation.

Selection and Effect of Ointment Bases for Preparing Collagenase Inhibitor Ointment Using High-Performance Liquid Chromatography and Franzcell Apparatus

After the dorsal subcutaneous administration of injectable collagen implant derived from bovine dermis (Zyderm; INAMED, Santa Barbara, CA) to mice, ointments that contain 3 types of collagenase inhibitors, Esculetin (6,7-dihydroxy-2H-chromen-2-one), ONO-4817 [(2S,4S)-N-hydroxy-5-ethoxymethyloxy-2-methyl-4-(4-phenoxybenzoyl) amino-pentanamide], and MMI270 (CGS27023A) {N-hydroxy-2(R)-[(4-methoxysulfonyl)-(3-picolyl)-amino]-3-methylbutanamide hydrochloride monohydrate} were applied daily on the dorsal region of mice (injection site), and intradermal Zyderm was extirpated after 30, 60, and 90 days to measure the level of hydroxyproline. Furthermore, dermal tissue was examined by Azan staining and immunostaining. A significant difference was observed in the level of hydroxyproline in the Esculetin and the ONO-4817 ointment groups compared with that in the control group after 30 days. A significant difference was also observed in the level of hydroxyproline in the Esculetin ointment group compared with that in the control group after 60 and 90 days. Histologically, 90 days after the application of the ointment, dense localization of type III collagen was observed around the injected Zyderm in the group applied Esculetin ointment compared with the control group. Therefore, it was indicated that Esculetin suppressed the degradation of collagen, and further facilitated the qualitative changes that increased neo-collagen, and that the collagen implant with hypodermic injection remained on behalf of ointments contained within the collagenase inhibitors that were applied on the skin surface.

Viscometric assay for determining lysozyme activity

Abstract A method was developed for determining the activity of lysozyme using two pipettes as a viscometer. The proposed viscometric assay uses carboxymethylchitin as a substrate and measures the enzyme activity at 22°C. The method requires only simple manipulations with the use of readily available laboratory equipment and the determination of enzyme activity is rapid.

The in vivo effect of esculetin ointment and esculetin-mixed Zyderm for Zyderm.

Background: Injectable collagen is often used for treatment of wrinkles or scars in cosmetic surgery. However, it is degraded within a short period after subcutaneous injection. The authors aimed to achieve a long-lasting effect of the filler with a new collagenase inhibitor, esculetin (6,7-dihydroxy-2H-chromen-2-one). Method: Nude mice were divided into two study groups and a control group (35 mg cattle collagen): (1) those implanted with Zyderm 0.3 g subcutaneously into the dorsal region followed by daily topical application of 5% esculetin ointment (0.5 g/day) to the skin of the implanted area (the 5% esculetin ointment group), and (2) those implanted with a mixture of Zyderm 0.3 g and esculetin (1 to 4 mM) (the esculetin-mixed Zyderm groups).1 In each group, Zyderm was removed at different time points to measure the wet weight and hydroxyproline level. Furthermore, each removed Zyderm specimen was sectioned for histologic examination with Azan staining and immunostaining. Results: In the esculetin ointment group and the 2 mM esculetin-mixed Zyderm group, the hydroxyproline levels at 30, 60, and 90 days were significantly higher than those in the control group, suggesting that esculetin suppresses the biodegradation of Zyderm. There was no significant difference in hydroxyproline level between the esculetin ointment group and the 2 mM esculetin-mixed Zyderm group; biodegradation occurred to a similar extent with either method of application. Conclusions: An atelocollagen implant is used as a safe and effective scaffold material for tissue regeneration. Future applications of the present study are expected.

Stability of Various Compositions in the Solution after being Excluded the Plastic Exterior Bag for TPN Kit and after being Broken Kit's Partition.

We evaluated the stability of various compositions after the plastic exterior bag of the TPN kit leaked out and the kits partition broke.PN-twin-2 and Aminotripa 2 leaked out of the plastic exterior bag in the double chambered type TPN kit, after being stored at room temperature for 14 days, a slight degree of coloration was noticed on the amino acid layer, while the A350 level (index for tryptophan degradation) was increasing.Coloration was evident after the kits partition broke, and the residual rate of glucose and amino acids were over 95%.As for Unicaliq N, the residual glucose and amino acid rates were also over 95%.

The suppressive effect of salicylazosulfapyridine on IL-6 production by human coronary artery endothelial cells.

It is thought that prominent production of inflammatory cytokines and expression of adhesion molecules on the endothelial cells activated by unkown pathogens induce the formation of the vasculitis in Kawasaki disease (KD) . To study the anti-inflammatory effect of salicylazosulfapyridine (SASP) on the cardiovasculitis of KD, we examined the effect of SASP on IL-6 production by human coronary artery endothelial cells (HCAEC) . We used HCAEC because IL-6 production by HCAEC was greater than by human umbilical vein endothelial cells. SASP at less than 0.5 mM suppressed IL-6 production by LPS-induced HCAEC, and higher concentrations of SASP affected cell viability. On the other hand, SASP did not suppress IL-6 production by the cells stimulated with cytokines such as TNFα, IL-1 β and IL-4. Sulfapyridine and 5-aminosalicylic acid, which are degradation products of SASP, did not suppress IL-6 production.These results show that SASP may have a direct inhibitoy effect on HCAEC activated by external substances such as LPS, which may be an effective agent in the cardiovasculitis of KD.

Secondary Contamination of Eyedrops.

We studied secondary contamination of eyedrops at our university hospital and Okinaga Ophthalmic Hospital.We found that 14 (2.77%) of 506 eyedrops containing 31 kinds of ophthalmic solutions recovered from outpatients (203), were contaminated.We did not find relationship between secondary contamination and a kind of antiseptic on eyedrops.

Variation by combination of hyperalimentation with fat emulsion (2)

Three types of fat emulsions were combined with hyperalimentation; the appearance, pH, and light microscopic observations on the mixture kept in a cool place at room temperature were made. The changes in the diameters of the fat particles were studied by Coulter nanosizer and the addition of the multiple vitamin causing the physical changes on the fat particles was also investigated. The enlargement of the diameters of the fat particles occurred due to the aggregation of the fat particles after mixing the emulsions with the hyperalimentation. A significant difference in stability of the mixture was observed between egg yolk lecithin and soybean lecithin when used as emulsifiers. The addition of the multiple vitamin inhivited the changes in the diameters of the fat particles, thus giving no hindrance in the usual clinical use. It was found that the fat soluble vitamins in the mixture worked as solubilizing agents to inhibit the chages in the diameters.

Effect of Ointments and Ointment Constituents on Oxidation Stability of Ergocalciferol

Stability of ergocalciferol (EC) in absorption ointment, hydrophilic ointment and seven kinds of ointment constituents were examined; in addition, antioxidative activities of gallic acid derivatives for EC in hydrophilic ointment were studied. Samples were exposed to air at 40°C and their oxidation stability was measured with high performance liquid chromatography (HPLC). Several kinds-of EC decomposition products were detected by HPLC. It was confirmed by the use of dual UV-wavelength that peaks corresponding to EC and other substances did not overlap each other.The results are as follows: 1) The amount of EC in hydrophilic ointment decreased ; however, that in absorption ointment did not change. 2) The amount of EC decreased markedly in lauromacrogol and sorbitan sesquioleate. 3) The amount of EC decreased as the proportion of stearyl alcohol in ointment increased, while cetanol showed reversible behavior. 4) The antioxidative activities of gallic acid derivatives for EC in hydrophilic ointment were as followes in the order of strength: lauryl gallate, isoamyl gallate, n-propyl gallate, ethyl gallate, gallic acid. The results of the experiment suggested that higher alcohol concentration is an important factor in the stabilization of EC in ointment.

Inquiries on Suspected Contents of Prescriptions at Four University Hospital Pharmacies

Details of inquiries on the suspected contents of the prescriptions were studied from February to April 1980 and from July to September 1981 at 4 university hospital pharmacies. By plotting a x-R control chart of the number of inquiries in each pharmacy, the changes of mean value and variance of the number could be checked quickly. This made possible to take a suitable and immediate action to these changes. Classified by department of consultation, the inquiries received at the internal department gave 40 to 50% of the total inquiries. Furthermore, the inquiries of drugs were about 60% in one pharmacy while the inquiries on amounts of drugs were about 50% in the other pharmacies. This implies that pertinent drug information on the inquiries in each pharmacy should be provided to physicians, primarily to internists.