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Background: This nationwide survey aimed to determine the status of jaundice management in Japan. Methods: A questionnaire about bilirubin level measurements and neonatal jaundice treatment was sent to 330 institutions providing neonatal care. The responses were analyzed according to institution level. Results: Of 330 institutions, 172 responded (52.1% response rate). Total bilirubin levels were measured in the central laboratory using spectrophotometry at 134 institutions and a blood gas analyzer at 81 institutions. Unbound bilirubin (UB) levels were measured by 79 institutions, while transcutaneous bilirubin measurements were taken at 63 institutions. There was no association between institution level and UB or transcutaneous bilirubin measurement. For phototherapy criteria, the Murata‐Imura criteria were adopted by 67 institutions, Nakamura criteria by 36, and Morioka criteria by 39. Light‐emitting diodes (LED) were used by 160 institutions versus fluorescent lights by 31. When a blue LED was used, 119 institutions used the high mode. There is no standard for increasing light intensity. No association was found between institution level and phototherapy criteria. UB was measured in 14 of 63 institutions using the Murata‐Imura criteria. Conclusions: There is a large variation in the management and treatment of neonatal jaundice among institutes in Japan. [ABSTRACT FROM AUTHOR]

Background Lycopene has been the object of considerable research attention recently, and the effects of the intake of lycopene, or of tomato products, have been studied in various ways. In Japan, interest in the health-promoting function of food components has increased. Objective Developing a method to determine lycopene contents in tomato that meets the Japanese Agricultural Standard (JAS). Method In the proposed JAS method, the test sample consists of fresh tomatoes; a hexane--acetone mixture is utilized as the extraction solvent. A collaborative study was conducted to evaluate the interlaboratory performance of the method. Results Ten laboratories participated and analyzed six test materials characterized by a lycopene content between 39 and 170 mg/kg as blind duplicates. After removing statistical outliers, RSDr ranged from 1.2 to 3.0% and RSDR ranged from 2.4 to 4.2%. The HorRat values were calculated and found to be in the 0.26--0.49 range. Conclusions The method for determining the lycopene content in tomato was evaluated by means of a collaborative study, and the reproducibility of this method was found to be acceptable. Highlights Intended for standardization in Japan, a method to determine lycopene content in tomato has been developed and shown to have acceptable precision in a collaborative study. [ABSTRACT FROM AUTHOR]

Phosphorus (P) plays important roles in aquatic ecosystems, but accurately determining phosphorus species (particularly bioavailable phosphate) is difficult. When phosphate concentrations are spectrophotometrically measured as soluble reactive P (SRP), the actual values are often overestimated. Ion chromatography is one method for accurately measuring orthophosphate concentrations. To clarify how the spectrophotometrical measurement overestimates actual phosphate concentrations, we compared estimates of phosphate concentrations in lake waters using both methods. SRP and orthophosphate concentrations in water samples collected from four shallow lakes around Lake Biwa, Japan, were determined monthly for 1 year by spectrophotometry and ion chromatography, respectively. SRP concentrations were frequently higher than those for orthophosphate in all lakes throughout the study period, suggesting that SRP and orthophosphate are not equivalent, although a significant relationship was found between them. Orthophosphate/SRP (ortho/SRP) ratios varied spatiotemporally in all lakes (range 0.11–1.04; mean 0.56), being well predicted by biological-mediated relevant parameters using a stepwise multiple logistic regression analysis (R2 = 0.76). The results implied that organic matter associated with biological activity contributes to the variability in ortho/SRP ratios. Such large variability in ortho/SRP ratios emphasizes the importance of accurate orthophosphate estimates for understanding P dynamics in aquatic ecosystems. [ABSTRACT FROM AUTHOR]

BACKGROUND: Capnometry detects hypoventilation earlier than pulse oximetry while supplemental oxygen is being administered. We compared the end-tidal CO2 (...) measured using a newly developed oxygen nasal cannula with a CO2-sampling port and the ... in extubated subjects after abdominal surgery. We also investigated whether the difference between ... and ... is affected by resting, by spontaneous breathing with the mouth consciously closed, and by deep breathing with the mouth closed. METHODS: Adult post-abdominal surgery subjects admitted to the ICU were enrolled. After extubation, oxygen was supplied at 4 L/min using a capnometry-type oxygen cannula. The breathing frequency, ... and ... were measured after 30 min of oxygen supplementation. PETro was continuously measured during rest, during breathing with the mouth consciously closed, and during deep breathing with the mouth closed. The difference between ... and ... during various breathing patterns was analyzed using the Bland-Altman method. RESULTS: Twenty subjects were included. The bias ± SD (limits of agreement) for breathing frequency measured by capnometry compared with those obtained by direct measurement was 0.4 ± 3.6 (-6.7 to 7.4). In ... compared with ..., the biases (limits of agreement) were 14.8 ± 8.2 (-1.3 to 30.9) at rest, 10.2 ± 6.4 (-2.3 to 22.7) with the mouth closed, and 7.7 ± 5.6 (-3.2 to 18.6) for deep breathing with the mouth closed. ... determined using the capnometry device yielded unreliable and widely ranging values under various breathing patterns. However, deep breathing with the mouth closed decreased the difference between ... and ..., as compared with other breathing patterns. CONCLUSIONS: ... measurements under deep breathing with mouth closed with a capnometry-type oxygen cannula improved the prediction of the absolute value of ... in extubated post-abdominal surgical subjects without respiratory dysfunction. [ABSTRACT FROM AUTHOR]

Background Persistent patent ductus arteriosus (PDA) is a frequent complication in preterm infants. Single nucleotide polymorphisms (SNP) in several genes, including angiotensin II receptor, type 1 ( AGTR1), transcription factor AP-2 beta ( TFAP2B) and tumor necrosis factor receptor-associated factor 1 ( TRAF1), have been reported to be associated with PDA in preterm infants. The aim of this study was to evaluate the relationships between PDA in preterm infants and polymorphisms in AGTR1, TFAP2B and TRAF1 in the Japanese population. Methods The subjects consisted of 107 preterm infants with gestational age <32 weeks. Extremely low-birthweight infants were treated with prophylactic indomethacin during the first 24 h after birth. Five SNP, namely, rs5186 in AGTR1, rs987237 and rs6930924 in TFAP2B, and rs1056567 and rs10985070 in TRAF1, were genotyped using TaqMan SNP genotyping assays. Results There were no significant differences in the distributions of the genotypes and allele frequencies of all studied SNP between the PDA group (n = 46) and the non-PDA group (n = 61). Conclusions There were no significant associations between the studied SNP and the incidence of PDA in Japanese preterm infants. These SNP may not be clinically important predisposing factors for PDA in Japanese preterm infants. [ABSTRACT FROM AUTHOR]

Background: In order to keep up the optimal iron status in chronic hemodialysis patients, it is important to know how much iron is lost due to hemodialysis. Residual blood associated with the hemodialysis procedure together with blood sampling inevitably causes the loss of iron in chronic hemodialysis patients. Recent advances in hemodialysis techniques might have reduced this complication. In this cross-sectional study, we directly measured total iron loss by hemodialysis.Methods: Two hundred thirty-nine patients who received chronic hemodialysis at Otowa Memorial Hospital were enrolled; 65.7% of patients were men, and mean age was 67 ± 6.4 years (mean ± SD) and 43.2% were diabetic. Residual blood in blood tubing set and dialyzer after rinse back with saline was collected and homogenized. The iron content including free, protein-bound and heme iron was measured using an atomic absorption spectrometry.Results: The mean iron content in residual blood was 1,247.3 ± 796.2 µg (mean ± SD) and the median was 1,002 µg (95% CI 377.6-3,461.6 µg), indicating 160.8 mg (95% CI 58.9-540.0 mg) iron loss annually when hemodialysis was performed 156 times a year. Fifty milliliter whole blood for monthly blood test and another 2 ml of whole blood lost by paracentesis at every dialysis session contains 228.6 and 118.9 mg iron at 11 g/dl hemoglobin, respectively. Therefore, an annual total iron loss due to hemodialysis comes to 508.3 mg (95% CI 406.4-887.5 mg).Conclusions: Five hundred milligram of annual iron supplementation might be sufficient to maintain iron status in hemodialysis patients, which is less than the dose recommended as 1,000-2,000 mg a year. Further study will be required to verify this iron supplementation dosage with recent hemodialysis procedure. [ABSTRACT FROM AUTHOR]

Background and Objective Biofilm formation occurs through the events of cooperative growth and competitive survival among multiple species. Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans are important periodontal pathogens. The aim of this study was to demonstrate competitive or cooperative interactions between these two species in co-cultured biofilm. Material and Methods P. gingivalis strains and gingipain mutants were cultured with or without A. actinomycetemcomitans. Biofilms formed on glass surfaces were analyzed by crystal violet staining and colony counting. Preformed A. actinomycetemcomitans biofilms were treated with P. gingivalis culture supernatants. Growth and proteolytic activities of gingipains were also determined. Results Monocultured P. gingivalis strains exhibited a range of biofilm-formation abilities and proteolytic activities. The ATCC33277 strain, noted for its high biofilm-formation ability and proteolytic activity, was found to be dominant in biofilm co-cultured with A. actinomycetemcomitans. In a time-resolved assay, A. actinomycetemcomitans was primarily the dominant colonizer on a glass surface and subsequently detached in the presence of increasing numbers of ATCC33277. Detachment of preformed A. actinomycetemcomitans biofilm was observed by incubation with culture supernatants from highly proteolytic strains. Conclusion These results suggest that P. gingivalis possesses a competitive advantage over A. actinomycetemcomitans. As the required biofilm-formation abilities and proteolytic activities vary among P. gingivalis strains, the diversity of the competitive advantage is likely to affect disease recurrence during periodontal maintenance. [ABSTRACT FROM AUTHOR]

Abstract: Introduction: The aim of this study was to compare cyclic fatigue resistance of four nickel-titanium rotary systems and to evaluate their surface, fractographic, and matrix morphology. Methods: Four models of endodontic rotary files (EasyShape [Komet/Gebr. Brasseler, Lemgo, Germany], ProTaper [Dentsply Maillefer, Ballaigues, Switzerland], NRT [MANI Inc, Toshigi-Ken, Japan], and AlphaKite [Komet/Gebr. Brasseler]) were subjected to fatigue testing in artificial canals with angle of curvature of 45° and 60° and a radius of curvature of 5 mm until fracture occurred. Nickel-titanium (NiTi) alloy properties were investigated by light microscopy, environmental scanning electron microscopy (ESEM), and energy dispersive x-ray spectrophotometry (EDS). ESEM analysis was conducted on new files to examine surface characteristics and on fractured samples to identify the crack origin and the fractographic features. Results: Analysis of variance testing revealed significant differences (P < .001) among the groups. NRT files had the highest fatigue resistance followed by AlphaKite, EasyShape, and ProTaper. All the new files presented surface imperfections. Fractographic analysis found the crack initiation to originate at the level of surface irregularities. Optical microscope inspection of the NiTi alloy matrix disclosed different-sized nonmetallic inclusions among models. EDS analysis of these inclusions showed that they were composed of carbon and oxygen in addition to nickel and titanium. Under light microscopy, austenitic grains appeared larger near the handle and smaller near the tip in all instruments. Conclusions: NRT files presented the longest fatigue life. All samples showed surface irregularities and nonmetallic inclusions. Austenitic grains were smaller near the tip than near the handle. The angle of curvature was confirmed to influence the fatigue life of NiTi instruments. [Copyright &y& Elsevier]

The article discusses the effect of coffee intake on the concentration of hippuric acid (HA) in urine. According to the authors, HA is used for the estimation of exposure of toluene among the industrial workers in Japan, which can be affected by benzoic acid consumption. According to the results presented coffee intake in the workers who were not exposed to toluene increased their HA level and spectrophotometric analysis have confirmed the absence of benzoic acid in coffee.

Background: High incidence of gallbladder cancer (GBC) is reported from North India, with elevated concentrations of heavy metals in water and soil. This Indo-Japan collaborative study compared presence of heavy metals in gallbladder tissues., Methods: Heavy metal concentrations were estimated in Indian GBC and cholecystitis tissues and compared with Japanese GBC and cholecystitis tissues. Spectrophotometry was done for 13 Indian gallbladder tissues (8 GBC, 5 cholecystitis) and 9 Japanese (5 GBC, 4 cholecystitis). Transmission electron microscopy (TEM) thin foil element analysis was done in 10 Indian samples (6 GBC, 4 cholecystitis)., Results: Chromium, lead, arsenic and zinc were significantly high in Indian GBC compared with Japanese GBC. Chromium, lead and arsenic were significantly high in the Indian cholecystitis tissues compared to the Japanese. TEM of Indian tissues demonstrated electron dense deposits in GBC., Conclusion: Heavy metals- chromium, lead, arsenic and zinc are potential carcinogens in Indian GBC from endemic areas. This preliminary study links presence of heavy metals in gallbladder cancer tissues in endemic areas.

The most appropriate method to preserve Bufferin 81-mg tablets dispensed for unit-dose packaging in the hospital pharmacy was examined. The surface color change of the tablets was investigated over time by spectrophotometry, and the decomposition rate of aspirin was measured by high-performance liquid chromatography (HPLC). To overcome these, it was found that we can effectively prevent color changes and preserve the quality by maintaining the humidity as 55% or less, storage with drying agent in a plastic or aluminum pack. It was revealed that the color changes became greater and the decomposition rate became higher as time passed. Color changes markedly affect the patients' compliance, and are found to be a very important factor. It was considered that the clarity of the correlation between the color change and decomposition rate may contribute to a decrease in the number of tablets discarded before the expiration date., (Copyright 2010 Elsevier B.V. All rights reserved.)

Aim: The aim of this study was to analyse occlusal parameters and body variables to examine the factors accounting for masticatory performance of Japanese children in relation to adults., Methods: Sixty-six children and 24 adults chewed three pieces of a colour-changeable chewing gum, separately, for 30 s, 60 s, and 90 s. The colour change was measured at 30 s using L* a* b* colour space system. Biting force, occlusal contact areas, and occlusal pressure were recorded by Dental Prescale. Number of strokes, and the height and weight of the subjects were included in the statistical analysis., Results: Within each sample, Student's t-test revealed significant differences among a* values at the three chewing times in which the value of a* (indicator of red) increased as the length of chewing time increased (P < 0.05). Multiple regression analysis showed that the rate of mandibular movement in preschool children has a significant impact on masticatory performance (P < 0.05). In schoolchildren, the rate of mandibular movement, biting force, and body weight were significant factors (P < 0.05). In adults, occlusal contact area, biting force, and the rate of mandibular movement were the significant factors (P < 0.05)., Conclusion: Factors affecting masticatory performance differ according to the developmental stage of the individual.

Recently, more immediate and precise cultivar-identifying methods targeting the specific and/or introduced gene(s) have been put into practical use for various rice cultivars. However trustworthy and innovative the biotechnological analyses may be, DNA purity and quality do have unpredictable influences upon the identification. Extraction methods of rice DNA have hardly ever been compared in a comprehensive manner. In this study, we investigated extraction characteristics of three methods by using 10 rice cultivars and then examined template availability of rice DNAs thereby extracted. An UV spectrophotometric study with a view toward methods revealed three different facts: The Isoplant II kit method with inhibitor absorption yielded the most DNAs, the Takara kit method with magnetic trapping produced the best DNAs free from contaminative proteins, and the enzymatic digestion method exclusively with enzymatic digestions prepared the best DNAs free from contaminative polysaccharides. Moreover, with a view toward cultivars, an insignificant difference in yield was not entirely bore out, and some difference in cultivar might cause significant difference in yield; however, no significant difference in purity was found among the cultivars used. On the other hand, electrophoretic images of the DNAs from the same cultivars showed considerable differences in quality among the methods. Furthermore, the DNA extracts from certain brands of rice proved really available for cultivar identification by using polymerase chain reaction (PCR) related to sequence-tagged sites. Therefore, this study suggested that these extraction methods may be used as the situation demands and that the DNAs thereby extracted might work successfully even in cultivar-identifying PCRs.

A simple and sensitive spectrophotometric method for the determination of human serum albumin (HSA) was established based on the ternary complex-formation reaction of HSA with o-sulfophenylfluorone (SPF) as a xanthene dye and metal ion (niobium(V) and bismuth(III)) in the presence of a dispersion agent. This new method enabled the determination of HSA in the range of 1 - 15 microg/ml HSA by measuring the difference of the absorbance at 530 nm between HSA-SPF-metal ion and SPF-metal ion solutions. In the determination of HSA, this method is about 2-times more sensitive than the Pyrogallol Red-molybdenum(VI) method (PR method), which accounts for more than 80% of the quantification methods for urinary protein assays in Japan. There was no significant difference between the results obtained by the present method and the PR method for human urine samples. The binding process between the SPF-metal complex and HSA was studied by determining the binding parameters and the thermodynamic parameters.

The purpose of this study was to investigate the effect of cool exposure on lipolytic response during prolonged intermediate-intensity exercise in humans. Eight male subjects participated in this study; they performed 120-min cycle ergometer exercise at 60% of maximal oxygen uptake (VO2max) in a climatic chamber at 10 degrees C (C) and 30 degrees C (WH). There were no significant differences in oxygen uptake and respiratory exchange ratio between the two conditions during the prolonged exercise. Significant influences of cool exposure were observed in the changes in both heart rate and rectal temperature (p<0.01). Although cool exposure had no significant effects on plasma triglyceride, free fatty acid, and glycerol levels, changes in adrenaline and noradrenaline levels at C were significantly lower than WH during the prolonged exercise (p<0.01). Changes in the ratio of glycerol to noradrenaline (Gly/Nad), as an index of lipolytic efficiency, were significantly high at C as compared with WH (p<0.01). These results suggest that cool exposure has an influence on lipid metabolism during prolonged intermediate-intensity exercise, from the viewpoint of efficiency in lipolysis.

The concentrations of D- and L-lactate, methylglyoxal and pyruvate were measured in tissues of normal and starved Octopus ocellatus. D-Lactate was always more abundant than L-lactate in the tissues. D-Lactate, pyruvate and methylglyoxal were present in 320, 94 and 43 times higher concentrations in tentacle of O. ocellatus of control group than those in normal rat skeletal muscle. The D-lactate concentration in the tentacle of O. ocellatus was 17-fold higher than that in Octopus vulgars. The activities of enzymes involved with D-lactate metabolism such as pyruvate kinase, octopine dehydrogenase, glyoxalase I and II and lactate dehydrogenase were measured in those tissues. The activities of glyoxalase I and II, and D-lactate dehydrogenase were increased in mantle and tentacle of starved octopus, while the levels of D-lactate and related metabolites were lowered in these tissues. The experimental results presented in this report and up to the present indicate that D-lactate is actively used for energy production in the tentacle and mantle of the starved animals. In octopus, especially starved octopus D-lactate was actively produced from methylglyoxal, which is formed via aminoacetone from threonine and glycine., (Copyright 2005 Wiley-Liss, Inc.)

We purified an extracellular hemoglobin with the molecular mass of ca. 440 kDa from the whole homogenates of Oligobrachia mashikoi (phylum Pogonophora) by a one-step gel-filtration. The preparation was pure to be crystallized. The P50 values of the hemoglobin and the fresh blood prepared from O. mashikoi were about 0.82 Torr and 0.9 Torr, respectively, which were much lower than the P50 value of human hemoglobin. However, the n values of the hemoglobin and the blood were about 1.2 and 1.1, respectively. Using the improved tricine SDS-PAGE, we could separate O. mashikoi hemoglobin into four kinds of the globin chains, A1, A2, B1 and B2, and succeeded for the first time in cloning and sequencing of the complete cDNA encoding B1 globin gene, in addition to A1, A2 and B2 globin genes in full length. We found that all globin genes have the extracellular signal sequences in each molecule and the distal His of the B1 globin chain is replaced to Gln. Finally, we constructed phylogenetic trees of the hemoglobins from Pogonophora, Vestimentifera and Annelida.

Objective: The daily bisphenol A (BPA) intake level of Japanese pregnant women was surveyed based on the measurement of the urinary excretion level of a BPA metabolite., Methods: Spot urine samples were collected from 56 pregnant women who visited the gynecology division of a hospital for a routine health check between June and October 2003. The urinary concentrations of the BPA metabolite and creatinine were measured by GC/MS/MS and spectrophotometry, respectively. Daily BPA intake was assumed to be equal to daily excretion., Results: The daily intake of BPA among Japanese pregnant women was estimated to be in the range of < 0.3 to 7.9 microg/day (median < 2.0 microg/day), being consistent with the levels in previous studies for non-pregnant Japanese women. This level was far below the current Acceptable Daily Intake (0.01 mg/kg/day) which was set by the European Commission. The maximum estimated intake per body weight (0.16 microg/kg/day) reached 1/10 of the Lowest Adverse Effect Level of BPA for pregnant mice for a reproductive effect on the offspring (2 microg/kg/day)., Conclusion: It is desirable to lessen BPA intake from a precautionary viewpoint, particularly in pregnant women.

This study was conducted to estimate the requirements of calcium (Ca), magnesium (Mg) and phosphorus (P) in Japanese young adults. From 1986 to 2000, 109 volunteers 123 males, 86 females), ranging from 18 to 28 y old, took part in mineral balance studies after written informed consent was obtained. The duration of the study periods ranged from 5 to 12 d, with 2-4 d of adaptation. Foodstuffs used in each study were selected from those commercially available. The minerals present in diet, feces, urine and sweat were measured by atomic absorption spectrophotometer (Ca, Mg) or spectrophotometer (P). The dietary intakes of Ca, Mg and P ranged from 4.83-23.58, 2.44-7.83 and 13.46-45.69 mg/ kg BW/d, respectively. Dietary intake (Intake) of Ca was positively correlated to apparent absorption (A.A.) (r2 = 0.425), which was also correlated with urine excretion (Urine) (r2 = 0.327) and balance (Bal) (r2 = 0.382). Intake of Ca was slightly but significantly correlated with Bal (r2 = 0.036, p = 0.048). The mean value and upper limit of the 95% confidence interval for the regression equation between Intake and Bal when balance is equal to zero (Mean and upper limit) for Ca were 11.752 and 12.555 mg/kg BW/d, respectively. Intake of Mg was positively correlated to A.A. (r2 = 0.451), which was also correlated with Urine (r2 = 0.486) and Bal (r2 = 0.349). However, Intake of Mg was not correlated with Bal. Intake of P was positively correlated with A.A. (r2 = (0.959), which was also correlated with both Urine (r2 = 0.908) and Bal (r2 = 0.135). Intake of P was slightly but significantly correlated with Bal (r2 = 0.103, p = 0.0013). Mean and upper limits for P were 22.584 and 24.059 mg/kg BW/d, respectively. Intakes of Mg and P correlated negatively with their respective A.A. rates (%) (r2 = 0.120 for Mg, r2 = 0.109 for P). However, there was not much of a correlation for Ca. Balance of Ca was positively correlated with that of Mg (r2 = 0.541), but not with that of P.

The uptake of plutonium(VI) and uranium(VI) by lichen biomass was studied in the foliose lichen Parmotrema tinctorum to elucidate the migration behavior of Pu and U in the terrestrial environment. Pu and U uptake by P. tinctorum averaged 0.040+/-0.010 and 0.055+/-0.015 g gdry (-1), respectively, after 96 h incubation with 4.0 x 10(14) mol 1(-1) Pu solutions of pH 3, 4 and 5. SEM observations showed that the accumulated Pu is evenly distributed on the upper and lower surfaces of P. tinctorum, in contrast to U(VI), which accumulated in both cortical and medullary layers. UV/VIS absorption spectroscopy demonstrates that a fraction of Pu(VI) in the solution is reduced to Pu(V) by the organic substances released from P. tinctorum, and the accumulated Pu on the surface is reduced to Pu(IV), while U(VI) keeps the oxidation state of VI. Since the solubility of Pu(IV) hydroxides is very low, reduced Pu(VI) does not penetrate to the medullary layers, but is probably precipitated as Pu(IV) hydroxides on the cortical lichen surface. It is concluded that the uptake and reduction of Pu(VI) by lichens is important to determine the mobilization and oxidation states of Pu in the terrestrial environment.

The effect of pH on Cu(I) and Cu(II) forms of the isolated soluble domain of the stellacyanin from Rhus vernicifera (SCu), the Japanese lacquer tree, has been studied by electronic and NMR spectroscopy and using direct electrochemical measurements. A pK(a) value of 10.1-10.4 is observed for the alkaline transition in this oxidized phytocyanin and results in a slightly altered active-site structure, as indicated by changes in the visible and paramagnetic (1)H NMR spectra. Electrochemical studies show that the pK(a) value for this transition in SCu(I) (reduced SCu) is 11.0. These results are compared with those recently obtained for other phytocyanins, and in particular umecyanin. In all cases, the alkaline transition is caused by the deprotonation of the surface lysine residue adjacent to the axial ligand. This lysine residue is completely conserved in known phytocyanin sequences. Also highlighted in these studies are the remarkable active-site similarities between stellacyanin and umecyanin.

A speciation scheme of trace molybdenum was proposed for river water based on size fractionation by filtration and ultrafiltration and the catalytic spectrophotometric determination of the reactive molybdenum concentration (CR). The total concentration (CT) of molybdenum was determined by the same method after acid decomposition to obtain the concentration (CT - CR) of unreactive molybdenum. Most molybdenum in natural river-water samples was found to be reactive species. A large part of the molybdenum was found in the fraction of molecular weight (MW) < 10(3), and was estimated to be MoO4(2-) from the chemical equilibria of molybdate ions. The residual part of molybdenum was found in the colloidal and particle fractions (MW > or = 10(4)), and was characterized as reactive molybdenum adsorbed or complexed on humic iron aggregates. The coexistence of silicate contributed to a decrease of the particle size of humic iron aggregates associated with molybdenum. The above-mentioned speciation results were confirmed by an analysis of artificial samples. The changes in the fractionation results by acidification (0.1 M HCl) were also used to characterize molybdenum in natural water.

An unidentified bacterial strain, SCRC-21406, isolated from the intestine of a marine fish, Glossanodon semifasciatus, produced docosahexaenoic acid at 23% (mol/mol) [= 28% (w/w)] of total fatty acids in a medium containing 0.5% (wt/vol) peptone and 0.1% (wt/vol) yeast extract at 12 degrees C under atmospheric pressure. The cell yield was 0.43 g/L. The major lipids of the strain were phosphatidylethanolamine and phophatidylglycerol. Docosahexaenoic acid was localized at the sn-2 positions of both phospholipids. The amounts of polyunsaturated fatty acids other than docosahexaenoic acid were extremely small [< 3% (mol/mol)]. Monounsaturated fatty acids of the cis-7, cis-9 and cis-11 types were detected.

Folic acid was tested for the preparation of "Folic Acid Reference Standard (Control 921)". The quality of the raw material was examined and compared with the previous Folic Acid Reference Standard (Control 862). Analytical data obtained were as follows: water content, 7.52%; free amines, 0.1%; infrared spectrum, the same as that of the JP Reference Standard; thin-layer chromatography, three impurities were detected; high-performance liquid chromatography (HPLC), two impurities were detected; assay, 101.2% by spectrophotometry at 550 nm and 100.4% by HPLC. Based on the above results, the raw material was authorized as the Japanese Pharmacopoeia Reference Standard (Control 921).

The "Acid Red Standard (C.I. 45100)" for the Dye Standard of the National Institute of Hygienic Sciences was prepared. The content of this Dye Standard determined by the titanium trichloride method was 96.4% on the average. The ultraviolet-visible absorption and infrared spectra of the Dye Standard were also measured.

The raw materials of d-camphor and dl-camphor were examined for preparation of the "d-Camphor Reference Standard" and "dl-Camphor Reference Standard". Analytical data obtained were as follows: ultraviolet spectrum, lambda max = 290nm; infrared spectrum, 2958, 1742, 1045cm-1; optical rotation, [alpha]D20 = +42.7 degrees (d-camphor), [alpha]D20 = -0.3 degrees (dl-camphor); melting point, 180 degrees C (d-camphor), 179 degrees C (dl-camphor); gas-chromatography (GC), one impurity was detected in d-camphor and three impurities in dl-camphor; assay, 99.5% (d-camphor), 99.5% (dl-camphor) by GC. Based on the above results, these raw materials were authorized as the Japanese Pharmacopoeia Standard (Control 911).

The "New Coccine Standard (C.I. 16255)" for the Dye Standard of the National Institute of Hygienic Sciences was prepared. The content of this Dye Standard determined by the titanium trichloride method was 97.5% on the average. The ultraviolet-visible absorption and infrared spectra of the Dye Standard were also measured.

The raw material of pyridoxine hydrochloride was examined for preparation of the "Pyridoxine Hydrochloride Reference Standard". Analytical data obtained were as follows: loss on drying, 0.00%; pH, 2.87; melting point, 204.6 degrees C (decomposition); infrared spectrum, same as Pyridoxine Hydrochloride Reference Standard (Control 879); thin-layer chromatography, nonimpurities were detected; assay, 100.0% by non-aqueous titration and 100.0% by UV spectrophotometry. Based on the above results, the raw material was authorized as the Japanese Pharmacopoeia Standard (Control 911).

Methyldigoxin was tested for preparation of the "Methyldigoxin Reference Standard (Control 881)". Analytical data obtained were as follows: infrared spectrum 3456, 1738, 1086 cm-1; ultraviolet spectrum lambda max = 219 nm; absorbance E1cm1% (219 nm) = 193.1; water 0.99%; optical rotation [alpha]546.1(20) = +22.5 degrees; thin-layer chromatography and high-performance liquid chromatography indicates two impurities, respectively; quantitative analysis of acetone 3.87%; assay by spectrophotometry 100.3% against Digoxin Reference Standard (Control 802). On the basis of the above results, this material was authorized as the National Institute of the Hygienic Sciences Reference Standard (Control 881).

The raw material of mestranol was examined for preparation of the "Mestranol Reference Standard". Analytical results for the sample were as follows: UV spectrum indicated absorption maxima at 279 and 287 nm and absorptivity at 279 nm E1%1cm 66; IR spectrum indicated specific absorption at 1612, 1578, 1505, 1253, and 1060 cm-1; optical rotation +3.9 degrees; melting point 153.1 degrees; loss on drying 0.03%; TLC and HPLC analyses indicated one impurity, respectively; the purity was assumed to be 99.3% by HPLC analysis. Based on the results, the present raw material was authorized to be the Reference Standard of the National Institute of Hygienic Sciences.

Some biochemical properties of a new red cell human carbonic anhydrase variant, CA Ih Hiroshima, have been determined. Evidence is presented that the amino acid substitution in the Japanese variant is not the same as the previously characterized CA Ic variant from Guam of similar electrophoretic mobility. Based on a comparison with the normal CA I isoenzyme, a proposal for the site of the amino acid substitution is presented.

During the course of a survey, a new hemoglobin, designated hemoglobin Yoshizuka, has been encountered in a Japanese family. Clinically, mild anemia was noted in five of six heterozygous individuals but no other significant abnormalities were found. Hemoglobin Yoshizuka is characterized by the substitution of aspartic acid for asparagine at the tenth residue of the G helix in the beta-chain. Reduced oxygen affinity with almost normal heme-heme interaction was found to be a property of this abnormal hemoglobin. The asparagine residue G10(108)beta lies in the internal cavity of the tetrameric molecule and its main chain carbonyl is thought to be hydrogen bonded to histidine G10(103)alpha at the region of contact between alpha- and beta-chains. It would appear likely that the introduction of a carboxyl group into the central cavity might result in interactions between the polar groups and the substituted side chain, disrupting the system of hydrogen bonds which contribute to the stability of the contacts between unlike subunits.