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Changes in color parameters (L*, a*, b*, and a*/b* values), carotenoid content, and singlet oxygen absorption capacity (SOAC) value were investigated at ripening of 'Matsuribayashi777', a large red-fleshed watermelon cultivar. The L* value decreased and the a* value increased up to 30 days after pollination, indicating that the flesh color turned reddish black rapidly, and the L* value increased from 40 to 50 days after pollination, indicating a slight fading of the color. The carotenoid content increased from 20 to 30 days after pollination, was high at 30 to 40 days after pollination, and decreased at 50 days after pollination. β-carotene content increased with days after pollination. The SOAC value increased from 20 to 30 days after pollination, with the highest value (12.55 µmol α- tocopherol equivalent/mL) at 30 days after pollination, and decreased at 50 days after pollination. Single correlation analysis of carotenoid content and color parameters showed a correlation of 0.898 between lycopene content and a* value. The carotenoid content has been reported to be closely related to SOAC value, and the same trend was confirmed in this study. The correlation between a* values and SOAC value at 20 to 40 days after pollination was 0.940, y = 0.9761x-13.626 (R² = 0.8838). These results suggest that the SOAC value of 'Matsuribayashi777' can be predicted from the fruit juice color measurement. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Information Science & Technology Association/Joho no Kagaku to Gijutsu is the property of Information Science & Technology Association and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of the Japan Research Association for Textile End-Uses / Sen'i Seihin Shohi Kagaku is the property of Japan Research Association for Textile End-Uses / Sen'i Seihin Shohi Kagaku and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The conventional analysis method has problems with extraction efficiency, operability, and reproducibility. In this study, we attempted to solve these problems and improve the analytical method to obtain sufficient extraction efficiency and good operability and accuracy. The conventional method was able to get sufficient extraction in dried meat products, where the extraction efficiency of the conventional method was low, by increasing the concentration of sodium hydroxide solution at the time of homogenization. Suction filtration after adding the defoaming agent was added allowed for accurate volume adjustment. The turbidity of the extract caused by insufficient addition of zinc acetate solution was removed by increasing the amount of zinc acetate solution that was added. Turbidity caused by starch was removed by adding pancreatin. The RSD of the quantitative values was improved by adding sodium hydroxide solution and 80-90℃ water and immediately homogenizing. Furthermore, by changing the dilution factor of the extract solution in the colorimetric method, the inhibition of coloration by reducing substances was suppressed, and more accurate quantitative values could be obtained than with the conventional method. The recovery rate was 78.5-105% (RSD 0.7-5.8%), which was a good result. This method was considered to be a useful analytical method that can contribute to improving the inspection accuracy of nitrite ion analysis.

Color feature analysis was applied to the characterization of color textures of 24 kinds of fabrics with various colors and patterns by using CIELAB color space. Color measurements of these fabrics were done with a commercially available color scanner. X, Y, Z, L*, a*, b*, C*ab and hab of CIE color system were obtained for each pixel (i, j) in the image of color textures by respective measurements and by appropriate transformation of the values. Mean values for L*, C*ab and hab of all pixels (AVE-L*, AVE-C*ab and AVE-hab) were obtained as color information parameters. The angular second moment (ASM), contrast (CON), correlation (COR), entropy (ENT) and fractal dimension (D) statistically extracted from distributions of L* for each fabric, were measured as visual feature parameters. A semantic differential analysis has been made on the color texture according to following feeling scales: vivid-obscure, gaudy-quiet, striking-unnoticeable, hard-soft, strong-weak, warm-cool, heavy-light, light-dark, complex-simple, variable-invariable and homogeneous-heterogeneous. The relationship between their mean sensory values and the image information parameters was examined by the factor analysis. These results indicated that feelings from patterns related to visual feature parameters (ASM, CON, COR, ENT and D); heavy, hard and dark feelings related to AVE-L*; vivid, gaudy, striking and strong feelings related to AVE-C*ab; and warm feeling related to AVE-hab. [ABSTRACT FROM AUTHOR]

The article focuses on a study which examines the color reproducibility of heat-press ceramics through the use of a staining technique. The study uses three colors in measuring the color of ceramic specimens with a small area colorimeter. It reveals that the translucency of ceramic ingots and the color and thickness of staining porcelain are significant factors in reproducing the target color.

We carried out performance evaluation tests using clinical isolates in a modified yeast fungal drug susceptibility test kit ASTY plus caspofungin (CPFG) (Kyokuto Pharmaceutical Industry Co., Ltd., Tokyo, Japan). We used 51 strains of clinical isolates ( Candida albicans , 30 strains; C. dubliniensis , 2 strains; C. glabrata , 4 strains; C. tropicalis , 4 strains; C. guilliermondii , 4 strains; C. parapsilosis , 4 strains; C. krusei , 3 strains). In this evaluation, minimum inhibitory concentration values of clinical isolates using the control CLSI method (Yeast-like fungi FP; Eiken Chemical, Tokyo, Japan), ASTY method, and another method (Yeast-like fungus DP; Eiken Chemical) were compared. The drugs with a concordance rate of 90% or more within ±1 tube were CPFG, amphotericin-B, 5-flucytosine, voriconazole, and those with a concordance rate of 80% or more were micafungin, fluconazole, and itraconazole. The above results clarified that the ASTY method correlated highly with the CLSI method.

In 1985, a (1→3)-β-D-glucan assay based on the discovery of a (1→3)-β-D-glucan-sensitive coagulation factor in the horseshoe crab amebocytes was developed in Japan, as a test aid in the diagnosis of invasive fungal infections. After many clinical trials, it has now been incorporated in practice guidelines both in Japan and abroad for the diagnosis and management of invasive fungal infection, and is now available worldwide. In Japan, however, two commercial kits that differ widely in sensitivity to (1→3)-β-D-glucan are used; namely, the colorimetric and the turbidimetric assays. Thus, attention must be paid when comparing measurement values between the two tests. In most cases, one pg/ml of the latter corresponds to about 7 pg/ml of the former. This raises the cutoff value of the turbidimetric assay far higher than that of the colorimetric assay, raising concern about missing many true cases of invasive fungal infections. The detection limit of the turbidimetric method is equivalent to as high as 60 pg/ml if converted to the colorimetric measurement. Thus, early diagnosis will be difficult to make based on detection of a rise in the β-D-glucan level in the lower range of concentration. The kits used overseas also differ in sensitivity; thus, global standardization or harmonization will be required to establish inter-assay data comparability and to enhance the usefulness of the tests. In the future, genetic engineering may provide an alternative source of the coagulation pathway enzymes other than the horseshoe crabs, in accordance with the global trends in animal protection, and bring about new opportunities for standardization.

The purpose of this study is to state the clinical and colorimetric features of pigmented lesions on oral mucosa. Among 201 pigmented lesions on oral mucosa, there were 74 melanin pigmentations (36.9%), 72 exogenous pigmentations (35.8%), 28 pigmented nevi (13.9%), and 27 malignant melanomas (13.4%). Melanin pigmentations were found mainly on the gingiva and lip, exogenous pigmentations were on the gingiva, pigmented nevi were found on the buccal mucosa, and malignant melanomas were on the upper gingiva. Thirty-six pigmented lesions were measured by colorimeter, and each lesion was evaluated numerically using the CIE L*a*b* colorimetric system. The statistical significance was confirmed by Tukey's test and a p value <0.05 was assumed as a significant difference. The color distribution of melanin pigmentation and exogenous pigmentation and malignant melanoma were significantly lower than normal mucosa in L*. A statistical significance was found between each pigmented lesion and normal mucosa in a* and b*. Among malignant melanoma and melanin pigmentation or exogenous pigmentation, a statistical significance was found in a* and b*. The color difference between melanin pigmentation and malignant melanoma was significant. The color distribution is thought to be different in each pigmented lesion, and we suggest that it is useful to use colorimetric features to diagnose each pigmented lesion.

Marshall and Warren were the first to succeed in culturing Helicobacter pylori (H. pylori) from the gastric mucosa of patients with gastritis in 1983. H. pylori is a spiral-shaped bacterium that resides in the gastric mucosa and is one of the most common infections worldwide. H. pylori infection causes gastritis and peptic ulcers and is associated with the development of gastric cancer and MALT lymphoma. Now, a variety of accurate diagnostic tests are widely available. Both invasive tests (bacterial culture, histopathology, and RUT) and non-invasive tests (UBT and serological test) are conducted for the diagnosis of H. pylori infection. This review provides a general overview of the diagnostic methods and tests the characteristics (sensitivity and specificity) for H. pylori infection.

We investigated the effect of aging on preoperative oxidative stress by measuring serum hydroperoxide values in 35 patients who underwent elective surgery under general anesthesia. A small amount of blood was withdrawn through an intravenous catheter and serum hydroperoxide values were measured using the Free Radical Analytic System (FRAS4). Aging and oxidative stress are significantly and positively correlated (P < 0.001, r = 0.622). Although the reason is unknown, oxidative stress increases concomitantly with aging.

Smoking has various influences on oral mucosa, including color change. We quantitatively analyzed color changes of oral mucosa by smoking. To examine the influence of smoking over a wider age range, we measured the color of oral mucosa of 62 nonsmokers and 56 smokers (age 30-83 years). We measured color with a colorimeter by a perpendicular course, illuminating the measuring sites from a distance of 1 meter by artificial sunlight. Before measuring, we used standard white for calibration. We used the CIE L * a * b * colorimetric system for the color specification. The measured sites were the lips, gingiva, tongue and buccal mucosa. Before colorimetry, we examined the precision and repeatability of colorimeter, but the change was small. Moreover, we similarly examined the sex difference and age difference, but there was no significant difference. This study was approved by the Tokyo Medical and Dental University ethics screening committee. Among nonsmokers and smokers, a statistical difference was recognized in lips L *, a *, gingiva L *, a *, and buccal mucosa L *, b *. Smoking decreased the luminosity of the lips, gingiva, buccal mucosa and caused a blue coloration of gingiva and buccal mucosa.

Salivary glands from 29 species of marine carnivorous gastropods in nine families were examined for lethal activity against mice and tetramine content. Mouse lethality was assayed by intravenous injection of buffer extracts into mice, and was detected in 14 species. Heat-stability tests confirmed that toxins in four species were thermolabile, while those in eight species were thermostable. Based on the tetramine contents determined by the colorimetric method using methanolic extracts, the thermostable toxins in seven species (Neptunea eulimatalamellosa, N. vinosa, N. arthritica, N. bulbacea, N. intersculpta f. pribiloffensis, N. intersculpta f. frater pilsbry and Hemifusus tuba) were considered to be tetramine contained at high levels (more than 900 micrograms/g salivary gland), but that in one species (Buccinum opisthoplectum) appeared to be a low-molecular-weight compound differing from tetramine. It is interesting that one (Hemifusus tuba) of the seven species containing high amounts of tetramine belongs to the family Melongenidae, although the other six Neptunea species are members of the family Buccinidae, as expected from previous studies.

I have observed discrepancies in the measurement of serum pancreatic lipase(LIP, Triacylglycerol acylhydrolase, EC 3.1.1.3) activities by non-enzymatic and enzymatic methods using diglyceride(1,2-dilinoleoyl-2-palmitoyl-glycerol, DLPG) as a substrate. The greatest discrepancies were seen in the post-heparin sera of patients with acute coronary syndrome(ACS). It is well-known that serum levels of lipoprotein lipase(LPL, EC 3.1.1.34) and hepatic triglyceride lipase(HTL, EC 3.1.1.3) show a significant increase after the administration of heparin in hemodialysis patients, and even larger quantities of heparin are administered before cardioangiography for ACS. Moreover, inhibitors of LPL and HTL induce almost complete reduction of increased LIP activity in post-heparin serum as measured by enzymatic assays. Pseudohyperlipasemia and positive interference from post-heparin serum LIP activity in enzymatic assays are considered to largely reflect LPL and HTL activities. Therefore, enzymatic reagent kits need improvement.

In recent years, we sporadically had some cases of accidental ingestion of drinks contaminated with detergent. In such cases it is necessary to identify the detergent for medical care or criminal investigation. Therefore, we made an attempt to establish a simple screening method of detergents, including household detergents, dishwashing detergents and shampoos using color reaction, thin-layer chromatography (TLC) and gel-permeation chromatography (GPC). In color reaction and TLC tests, it was possible to identify the individual sample. In the GPC test, several samples showed characteristic chromatograms, and even the same types of detergents did not give the same chromatogram patterns. From the above-mentioned results, it was suggested that this simple and rapid method may practically be useful to predict the contamination of detergent when controls are available, although not suitable for classification of detergent types.

The application of the endotoxin test for globulin and other blood products were investigated by two different limulus amebocyte lysate (LAL) test methods, colorimetric and kinetic turbidimetric methods, using two endotoxin-specific reagents. By the dilution of the blood products in 40 times or more, spiked endotoxin in the products was recovered accurately showing neither inhibition nor enhancement. The definite difference was not shown between the results obtained by the two LAL test methods. According to the method of the endotoxin test described under General Tests of The Japanese Pharmacopeia (thirteenth edition), JPXIII, the maximum valid dilution (MVD) for these products will be calculated to be 40 or more, so it is capable to measure the endotoxin limit for each product. This study indicates that the endotoxin test is applicable to measure the endotoxin content in globulin and other blood products as an alternative method for the rabbit pyrogen test.

Skin color of free flaps can represent their vascular conditions after reconstructive surgery. Thus far, however, surgeons have subjectively inspected flap color changes. More precise and objective measurement is necessary for early detection of vascular insufficiency. In order to evaluate color changes of flaps we used a portable non-contact type colorimeter with a L*a*b* color space (L* is brightness and a*, b* are chromaticness) recommended by the CIE (Commission Internationale d'Eclairage). Initially, we successfully performed objective evaluation of flap color changes in rats with or without a tied femoral artery or vein. Vein failure showed a decrease in a* and b* soon after surgery, while the artery failure appeared as b* and L* after 12 and 24 hours, respectively. Then, the color differences between sites of clinical free flaps were examined and considered to be relatively negligible against the time-dependent changes using delta a*, delta b*, delta L*, which represents the difference in color of an objective point from a control point. Next, we demonstrated that the color of abdominal skin did not significantly differ from that of forearm skin using delta a*, delta b*, delta L*. The clinical free flaps were measured with these parameters pre- and post-operatively and their normal course of color changes was revealed. The findings showed that a* changed with characteristic ups and downs before and after elevation due to congestion.

A new tetrazolium compound, WST-1, a sodium salt of 4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate, was introduced into clinical chemistry. The compound's dehydrogenase activities are useful in the colorimetric determination of the bioreducibility of cells. It proved to have a sensitivity similar to MTT, and produces a highly colored formazan dye under reductive conditions. Compared with the MTT assay, we found the WST-1 assay to be more sensitive, more convenient, and more exact. We used the new assay to study aging-related changes in human lymphocyte blastformation. Four groups of healthy adults aged 40-49 years (N = 40), 50-59 years (N = 40), 60-69 years (N = 40), 70-76 years (N = 40) were examined. Moreover, we studied lymphocytes from 16 patients (47-74 years) with probable Alzheimer's disease. For healthy controls, a negative correlation (r = -0.3108, p < 0.05) between age and enzyme activity was noted. The regression equation was: Y = -0.0085X + 1.473 where X and Y designate the age of the individual and enzyme activity (absorbance), respectively. On the other hand, for patients with Alzheimer's disease, age and enzyme activity did not correlate. These results indicate that the WST-1 assay may be useful in the study of changes associated with aging, and also more so than the MTT assay.

We determined in vitro interactions when amphoteric B (AMPH) is combined with various antimicrobial agents against yeast by using a colorimetric microdilution transfer plate technique, principally based on the current National Committee for Clinical Laboratory Standards (NCCLS) M27-T. An oxidation-reduction dye, sodium resazurin, was used as a color indicator to detect the growth of yeasts, and a standard two-dimensional, two-agent microdilution checkerboard in RPMI 1640 was employed to determine in vitro interactions; synergistic, indifferent or antagonistic. The study included 125 clinical isolates of Candida species and nine reference strains of American Type Culture Collection described in M27-T. Among the 34 antimicrobial agents first tested, polymyxin B (PL), rifampicin (RFP), tetracycline (TC) and erythromycin (EM) showed significant synergism. The fractional inhibitory concentration (FIC) indices of the respective agents were; PL 0.16 to 0.51 (mean, 0.315), RFP 0.13 to 0.56 (0.255), TC 0.06 to 0.75 (0.353) and EM 0.27 to 1.0 (0.550). RFP was the most potent agent, 118 of 125 clinical isolates (94.4%) showing synergism (FIC, < or = 0.5). With these results, we can conclude that several antibacterial agents are potentially effective when combined with AMPH against yeasts, probably due to alteration in the permeability barrier of the surface membrane by AMPH. Antifungal synergism may be promising for more effective, and less toxic therapy, and thus in vivo study will be necessary to determine their clinical significance.

A new colorimetric microdilution antifungal susceptibility testing using an air-dried microplate containing serially diluted antifungal agents and an oxidation-reduction dye, sodium resazurin, as a color indicator to detect the growth of yeasts was evaluated at multiple sites. Six ATCC reference strains described in the National Committee for Clinical Laboratory Standards (NCCLS) M27-T, two quality control and four reference strains, were repeatedly tested against five antifungal agents, amphotericin B(AMPH), flucytosine (5-FC), fluconazole (FCZ), miconazole (MCZ) and itraconazole (ITZ) at 17 different laboratories. Of 1,521 MIC determinations, 1,393 (91.6%) were within the acceptable limits described in the M27-T, ranging from 23.8% to 100%. However, significant discrepancies from the M27-T limits were noted in some combinations, such as FCZ against C. albicans ATCC 90028. In addition, the MIC limits for MCZ and ITZ against the six reference strains were determined. A total of 1,645 clinical isolates of yeasts were tested against five antifungal agents. Most isolates of C. albicans were highly susceptible to the five agents with MIC90 ranging from 0.03 microgram/ml (MCZ) to 1.0 microgram/ml (AMPH). The MICs against AMPH and 5-FC were tightly clustered, except with C. krusei and Trichosporon spp. Whereas, the distribution of MICs for FCZ, MCZ and ITZ had a wide range from < or = 0.03 microgram/ml to > 64 micrograms/ml. From these results, it can be concluded that our colorimetric broth microdilution antifungal susceptibility test is reliable, easy-to-perform and a suitable alternative for the determination of the MICs in clinical microbiology laboratories.

A colorimetric test method using the microplate culture technique for the determination of susceptibility of Mycobacterium tuberculosis against antimycobacterial agents was developed and evaluated by the multicenter study. The test method utilizes an oxidation-reduction dye, 2,3-diphenyl-5-thienyl-(2)-tetrazolium chloride (STC), as an indicator of mycobacterial growth. When compared to the presently available test method, some modifications were also included; lower inoculum density (10-fold dilution), inclusion of an inoculum diluted 1:100 as a growth control, and the preparation of inoculum preincubated in Middlebrook 7H9 broth and spectrophotometrically adjusted to McFarland #1 turbidity. The test method evaluated was highly precise and reliable to detect antimycobacterial resistances when the ATCC reference strains were tested. Also, the interpretations of the test result were highly comparable to those determined by the method of NCCLS M24-P, the % agreements ranging from 76.1% (ethambutol) to 91.3% (streptomycin). The test results were also comparable to those determined by Ogawa media; > 90% agreed with susceptible, intermediate, or resistant. The appearance of mycobacterial colonies on the test media was easily read, and the test results were more comparable to those of NCCLS M24-P. With these results, it can be concluded that the colorimetric microplate susceptibility test method described will be more suitable for clinical mycobacteriology laboratories.

A colorimetric broth microdilution modification of the National Committee for Clinical Laboratory Standards (NCCLS) for antifungal susceptibility testing was developed and evaluated. The test method modification includes; air-dried microdilution trays, in which serial two-fold dilutions of three antifungal agents, amphotericin B (AMPH), flucytosine (5-FC) and fluconazole (FCZ) were first prepared and evaporated, then reconstituted by adding 100 microliters of yeast inocula, and an oxidation-reduction color indicator (sodium resazurin, Sigma) added to RPMI 1640 medium buffered to pH7.0 with 0.165M morpholinepropanesulfonic acid. The trays were incubated in air at 35 degrees C and were inspected after 24 and 48hr of incubation. The MICs were defined as the lowest concentration of the respective agents; no color change (blue) for AMPH, and slight color change (blue to purple) for 5-FC and FCZ. The MICs of AMPH, 5-FC and FCZ were determined for four reference strains and 100 clinical isolates, in comparison with the NCCLS macrodilution method. The four reference strains were tested seven times each by macrodilution method (MACRO) and 14 times each against all three antifungal agents by microdilution method (MICRO). Overall, 100% of MICs determined by MACRO, 96% of those determined at 24hr incubation by MICRO, and 93% of those determined at 48hr incubation by MICRO fell within the 3-log2 dilutions, although 20 to 33% were out of the acceptable MIC ranges of the NCCLS M27-P proposal. Excellent reproducibility in determining growth endpoint by color change was also demonstrated, giving 99 to 100% agreements in duplicated dilutions. When the MICs determined for 100 clinical isolates against three antifungal agents were compared, those determined at 24hr incubation by MICRO gave 61.7% of agreement within the 3-log2 dilutions of the NCCLS MACRO, and those determined at 48hr incubation by MICRO were 86.2%. The MICRO read at 24hr trended to the lower MICs, except for 5-FC. While the MICs of MICRO read at 48hr were mostly comparable (78 to 97% agreement) to those of MACRO, especially against 5-FC (97%) and AMPH (85%), but the discrepant MICs of Candida tropicalis against FCZ were noted. With these results, it can be concluded that the resazurin colorimetric broth microdilution method is easy to perform and highly precise, and may provide MICs comparable to those determined by the reference NCCLS macrodilution method.

Enzyme immunoassay in measured its enzyme activity as a signal. This signal detection method have been progressing from colorimetric to fluorometric determination, and most recently amongst for more sensitive substrate chemiluminescent determination is popular. The methods, colorimetric determination is recognized to be less sensitive. We confirmed that the alkaline phosphatase labeled enzyme, cycling method significantly increased its sensitivity by changing the substrate. If the minimum detection limit is not sensitive enough by colorimetric determination, fluorometric is useful, and chemiluminescence can be applied when ultra sensitive method is required. We briefly described the characteristics of the main substrates which may very the sensitivity in these assays.

The method of enzyme activity related in enzyme immunoassay was described. The detection methods were colorimetric, fluorometric and chemi- or bioluminometric assay for enzyme, and the sensitivity of three detection methods were compared. The order of detection limit was chemi- or bioluminometry > fluorometry > colorimetry. But it is important that the selection of detection method of enzyme will make fit on your desirable sensitivity of analysis.

Reverse transcriptase (RT) inhibiting antibody in a series of plasma of HIV-1-seropositive subjects was quantitatively measured by poly A-linked colorimetric microtiter plate assay. The plasma were obtained from 6 asymptomatic carrier (AC)s and from 3 patients who progressed to AIDS. They had been followed 29-51 months. RT inhibiting antibody levels in the plasma were measured by inhibition assay against HTLV-IIIB RT activity. In five of the 6 AC cases, RT inhibiting antibodies in the serial plasma maintained high levels, and 50% inhibiting titers of the serial plasma did not decrease throughout the observation periods (45-51 months). HIV isolation from peripheral blood mononuclear cell (PBMC) of these 5 ACs did not succeed, and HIV p24 antigens were not detected in the plasma. In one AC case (046) RT inhibiting antibody levels gradually decreased after 48 months. In this case, HIV p24 antigen was not detected in the serial plasma throughout the observation period (48 months), but HIV was isolated from PBMC after 27 months. On the other hand, RT inhibiting antibody levels in the serial plasma of all 3 patients who progressed to AIDS gradually decreased in observation periods (29-35 months). HIV strains were isolated from these 3 cases. These results suggest that reduction of RT inhibiting antibody levels correlate well with the success of HIV isolation and with progression of clinical manifestation.

Polyvinylpolypyrroridone (PVPP) has been used in many countries for removing polyphenols in beer and wine, and was permitted as a food additive in Japan in April 1995. Prior to this approval of the compound, we studied procedures for identifying PVPP and its water soluble substances by colorimetry and infrared (IR) spectrometry for the establishment of the specification.

The role of M-CSF in the process of mammalian development has been drawing attention. Unlike mammalian embryonic development, in which M-CSF is supplied by the maternal body, avian embryonic development begins and proceeds under the influence of a certain amount of M-CSF present in each fertilized egg, and its concentration can be experimentally controlled while observing the embryo's developmental state. Apart from this fact, aves provide us with many other advantages in the study of embryonic development. It is, however, not easy to separate multipotent hematopoietic stem-cell fractions from avian bone marrow. In addition, because a macrophage colony forming assay in soft agar, though it has been widely used, requires a large number of cells and a long culture period, the method of assay is not adequate for the sensitive detection of M-CSF activity in small scale samples. We have established a method for the depletion of nucleated erythrocytes from chicken bone marrow cell suspensions using percoll density gradient centrifugation, and have also developed a new method for determining chicken M-CSF-like activity employing a liquid culture. In this method, a 100 microliters aliquot of fractionated hematopoietic stem cells, 1 x 10(5), was placed in a well of 96-well flat-bottom culture plate, 100 microliters of sample was then added to each well, and the uptake of neutral red was measured after 4 days of culture. These procedures represent a simple and sensitive means of detecting M-CSF-like activity in chicken serum of x60 to x32 dilutions.(ABSTRACT TRUNCATED AT 250 WORDS)

Plasma antithrombin III activity and concentration were measured simultaneously on COBAS MIRA-S by colorimetry using a synthetic substrate and by immunoturbidimetry, respectively, in normal subjects (n = 30), in patients with liver disease (n = 37), and in those with thrombosis (n = 76). The activity/concentration ratio of plasma antithrombin III, in which concentration was represented as the ratio to the mean concentration for normal, was evaluated in each group. Results showed that the ratio was significantly lower in patients with liver diseases (0.76 +/- 0.18, mean +/- SD, p < 0.01) and in thrombotic patients (0.89 +/- 0.18, p < 0.01) than that in normal (1.07 +/- 0.06), demonstrating the predominance of antithrombin III molecules with low activity in the former groups. This simple and rapid (< 10 min) method is applicable for detecting such the constitutional change in plasma antithrombin III.

The Limulus test, which has been adopted as the Test for Bacterial Endotoxins in the JP XII, can detect or quantitate endotoxins of Gram-negative bacterial origin using blood corpuscle extracts (Limulus amebocyte lysate, LAL) of horseshoe crabs (Limulus polyphemus, Tachypleus tridentatus, etc.). It may be conducted by the gel-clot or spectrophotometric (turbidimetric and colorimetric) techniques, the former being based on gel formation due to the activation of LAL by endotoxins. The turbidimetric technique is based on the LAL turbidity change during the gel formation and the colorimetric technique on activation of peptide hydrolytic enzymes in LAL. The Limulus test has been unofficially utilized as a simple and highly sensitive method for the determination of endotoxins in parenteral drugs in lieu of the in vivo Pyrogen Test using rabbits. For the Bacterial Endotoxins Test of the JP XII, the gel-clot technique alone was adopted, the technique being only allowed for Injection. Although most parenteral drugs show inhibition or enhancement in practice, this test can be most easily conducted by eliminating interfering effects through dilution of specimens by a factor not exceeding the maximum valid dilution (MVD) with water. Since MVD is dependent on the sensitivity of applied methodology, the turbidimetric and colorimetric techniques, which are more sensitive than the gel-clot technique, have a distinct advantage. The JP, as the leading Pharmacopoeia for the international harmonization of Bacterial Endotoxins Testing, has presented a "Draft towards International Harmonization of Bacterial Endotoxins Test", whose main purpose is the introduction of supplementary turbidimetric and colorimetric techniques. Under these circumstances the following subjects are discussed: (1) the proposal that, with a view towards international harmonization of the technical requirements of Pharmacopoeias, both the turbidimetric and colorimetic techniques should be included together with the gel-clot technique, (2) the differences in the testing principles and/or conditions prescribed in the current Bacterial Endotoxins Test of JP, USP and EP, and (3) the worldwide situation for Endotoxin Reference Standards.

The present work details an improved application of kit to the measurement of lipid peroxides in human serum. Kits based on the reaction of lipid hydroperoxides with a leucomethylene blue derivative in the presence of hemoglobin. Overall sensitivity was increased by taking samples 5 times larger than those indicated in product literature for the system. It was necessary, however, to compensate actual data for protein error. To do this, the following empirically obtained formula was employed: A = a/(-3.45x + 98) x 100, where A (nmol/ml) indicates compensated data for lipid peroxides, a (nmol/ml) indicates the data for the actual measurement of lipid peroxides, and x (g/dl) indicates the total protein. We obtained what we considered to be satisfactory results by the above formula, as it allowed us to observe that lipid peroxides in fresh human serum undergo a change, momentarily.

The stability of allantoin was studied in pH 3.0, 6.0, and 8.0 buffer solutions at 50 degrees C for 55 d. Allantoin was determined by high performance liquid chromatography (HPLC) with a cation exchange mode and colorimetry. The known degradation compounds, allantoic acid and glyoxylic acid, were simultaneously determined by ion chromatography. Urea was determined by HPLC with an ion pair mode. In the stability studies on pH 6.0 and 8.0 solutions, new degradation compounds of allantoin were identified as two growing unknown peaks in the HPLC chromatogram. Isolation of these compounds by preparative HPLC and 1H, 15N and 13C-NMR spectra revealed that they were condensates of allantoin and glyoxylic acid, which also reacted positively in phenylhydrazine colorimetry. The amounts of these compounds were calculated from the determination results of other allantoin related compounds in the buffer solutions and were estimated as the biggest amount among all other compound. As they were found to be stable in an alkaline solution, conditions for hydrolysis in the colorimetric determination of allantoin were further examined. It was found that the longer hydrolysis gave more than 90% recovery in the colorimetry.