Age-dependent changes in eumelanin composition in hairs of various ethnic origins

Hair pigmentation is one of the most conspicuous phenotypes of humans. From a chemical point of view, however, data remain scarce regarding human hair pigmentation characteristics. To determine melanin content and composition in human eumelanic hair from individuals of different ethnic origins and at different ages, we collected hair from 56 subjects with eumelanic hair from each group of African-American, East Asian, and Caucasian origin. The 56 subjects consist of 14, seven each of males and females, each from four age classes of younger than 11, between 12 and 19, between 20 and 45, and older than 46. We analysed hair colour scale, total melanin value, and contents of pyrrole-2,3,5-tricarboxylic acid (PTCA) and pyrrole-2,3-dicarboxylic acid (PDCA). We measured age-dependent increases in the relative quantity of eumelanin in pigmented human hairs in the three ethnic groups. Regarding melanin composition, we observed an increase in the PDCA/PTCA ratio with age in African-American and Caucasian hairs until approaching the quite constant level of the ratio in East Asian hairs in the elderly individuals. Our results evidence differences in the content and composition of eumelanin in human hair among African-American, Caucasian and East Asian individuals. Furthermore, we show evidence of age-dependent changes in the quantity and quality of eumelanin in pigmented human hairs. In particular, the age-dependent modification of the PDCA/PTCA ratio, a marker for 5,6-dihydroxyindole units in eumelanin, suggests a chronological evolution of hair follicle melanocyte phenotype (e.g. decrease in dopachrome tautomerase expression).     

Accumulative gene integration into a pre-determined site using Cre/loxP

Site-specific gene recombination systems, such as Cre/loxP, have been used for genetic modification of cells and organisms in both basic and applied research. We previously developed an accumulative gene integration system (AGIS), in which target gene cassettes could be repeatedly integrated into a pre-determined site on a plasmid or cellular genome by recombinase-mediated cassette exchange (RMCE), using Cre and mutated loxPs. In the present study, we designed a simplified AGIS. For gene integration into a target site, the previous system used two loxP sites in the acceptor DNA, whereas the new system uses a single loxP site. The gene integration reactions were repeated four times in vitro using Cre protein and specific plasmids. The expected integration reactions mediated by Cre occurred at the loxP sites, resulting in integration of four target genes. The system was also used for genomic integration of reporter genes using Chinese hamster ovary (CHO) cells. The reporter genes were efficiently introduced into the CHO genome in a Cre-dependent manner, and transgene expression was detected after the integration reaction. The expression levels of the reporter genes were enhanced, corresponding to the increase of transgene copy number. Recombinase-mediated AGIS provides a useful tool for the modification of cellular genomes.     

Selective production of lactic acid in continuous anaerobic acidogenesis by extremely low pH operation

The selective production of lactic acid by anaerobic acidogenesis with low pH control was examined using a chemostat culture. By decreasing culture pH to 3.5 in a chemostat culture containing mixed microbial populations for anaerobic acidogenesis, heterolactic fermentation became dominant, resulting in the selective production of lactic acid and ethanol. This phenomenon was reversible between the acidic and neutral conditions, and was not affected by the dilution rate. The extremely low pH operation was effective for selective lactic acid production in anaerobic acidogenesis.     

Lysophosphatic acid modulates prostaglandin secretion in the bovine uterus

Lysophosphatidic acid (LPA) modulates prostaglandin (PG) synthesis via LPA receptor 3 (LPAR3) in the murine endometrium. The lack of functional LPAR3 in mice may lead to embryo mortality. In the present study, we examined the role of LPA in the bovine uterus. We confirmed that LPA is locally produced and released from the bovine endometrium. Moreover, there are enzymes involved in LPA synthesis (phospholipase (PL) D(2) and PLA2G1B) in the bovine endometrium during estrous cycle and early pregnancy. Expression of the receptor for LPA (LPAR1) was positively correlated with the expression of PGE(2) synthase (PGES) and negatively correlated with the expression of PGF(2alpha) synthase (aldose reductase with 20 alpha-hydroxysteroid dehydrogenase activity - PGFS) during early pregnancy. In vivo LPA induced P4 and PGE(2) secretion was inhibited by LPAR1 antagonist (Ki16425). The overall results indicate that LPA is locally produced and released from the bovine endometrium. Moreover, LPAR1 gene expression in the endometrium during the estrous cycle and early pregnancy indicates that LPA may play autocrine and/or paracrine roles in the bovine uterus. LPAR1 gene expression is positively correlated with the expression of the enzyme responsible for luteotropic PGE(2) production (PGES) in endometrium. In cow, LPA stimulates P4 and PGE(2) secretion. Thus, LPA in the bovine reproductive tract may indirectly (via endometrium) or directly support corpus luteum action via the increase of P4 synthesis and the increase of PGE(2)/PGF(2)(alpha) ratio. It suggests that LPA may serve as an important factor in the maintenance of early pregnancy in cow.     

Amplified fragment length polymorphism of the AWA1 gene of sake yeasts for identification of sake yeast strains

Sake yeasts are used for sake brewing and have a crucial role in the quality of sake, since they produce not only ethanol but also various compounds that provide sake flavors. Therefore, the appropriate selection and monitoring of a strain used in sake mash is important. However, the identification of specific sake yeast strains has been difficult, because sake yeasts have similar characteristics in taxonomic and physiological analyses. We found amplified fragment length polymorphisms (AFLPs) in the PCR products of the AWA1 gene of sake yeast strains. The AWA1 gene encodes a cell wall protein that is responsible for foam formation in sake mash. This polymorphism of the AWA1 gene can be used for the identification of sake yeast strains.     

Myosin is solubilized in a neutral and low ionic strength solution containing l-histidine

Myosin, one of the major myofibrillar proteins, is insoluble at low and physiological ionic strength and soluble at high ionic strength. In this study, the behavior and morphology of myosin solubilized in a low ionic strength solution containing l-histidine (l-His) was investigated. More than 80% of myosin was solubilized in a low ionic strength solution with dialysis against a solution containing 1 mM KCl and 5 mM l-His. Transmission electron microscopy with rotary shadowing demonstrated that the rod of myosin in a low ionic strength solution containing l-His is longer than that of myosin in a high ionic strength solution. The elongation of the myosin rod in a low ionic strength solution containing l-His would inhibit the formation of a filament, resulting in the solubilization of myosin.     

Colorimetric microdetermination of sulphites in foods by use of the modified rankine apparatus. IV.

Summary Detection and determination of traces of sulphites in foods was attempted by use of the modified Rankine apparatus and pararosaniline colorimetry. Replacement of alkaline titration reported previously by pararosaniline colorimetry lowered the absolute detection limit from 30 g (titration method) to 2 g. In view of clean analysis, in the color developing system, 0.1 N-sodium hydroxide was used in place of mercuric chloride solution commonly used as an absorbant of sulphites. In order to prevent oxidative decomposition of sulphites during operation, nitrogen gas was used as carrier instead of air. Dimedone and sodium azide were used for the elimination of aldehydes and nitrites, respecitvely, in the sample, which will disturb the color development of sulphites with pararosaniline-formaldehyde reagents. With this improved method, it was possible to determine the residual sulphites in frozen peeled shrimps, sugared beans and other foods with low sulphite contents accurately.

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Colorimetrische Mikrobestimmung von Sulfiten in Lebensmitteln bei Anwendung der modifizierten IV. Rankine Apparatur

Zusammenfassung Geringe Sulfitmengen in Lebensmitteln (geschälte Garnelen, gezuckerte Bohnen) können colorimetrisch bestimmt werden. Die neuentwickelte Methode beruht auf einer Kombination von colorimetrischer Bestimmung mittels p-Rosanilin und der Bestimmungsmethode nach Rankine. Auf diese Weise lassen sich Gehalte von 2 g noch genau bestimmen. Bei der Farbentwicklung wurde das giftige Quecksilbertetrachlorid durch 0.1 n-NaOH ersetzt, anstelle von Luft Stickstoff als Trägergas verwendet und somit eine Oxydation des Sulfits während der Bestimmung vermieden. Da Nitrit und Aldehyde die Farbentwicklung stören, wurde ihr Einfluß durch Dimedon und Natriumazid ausgeschaltet.

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Studies on the Analyses of Sulphites in Foods (IV)     

Establishment of a modified rankine method for the separate determination of free and combined sulphites in foods. III

Summary Considerable improvements were made to the original Rankine method. Replacement of aspiration with an injection system contributed a great deal to the simplification of procedure, being accompanied with an increase in reproducibility. Air (flow rate 1.01/min) was used for injection because the use of inert gas gave little increase in recovery rate.Sodium bisulphite (free sulphite) and three kinds of combined sulphite compound (bisulphite adducts of acetaldehyde, pyruvic acid and D-mannose) were used to find the most suitable conditions for the separate determination of free and combine sulphites.Free sulphite was expelled from the sample by bubbling at 0 °C for 30 min. It was confirmed that no combined sulphite was dissociated under these conditions. The phosphoric acid concentration had an important role in the liberation of sulphite. When 25% phosphoric acid was used, more than 99% of free sulphite was expelled by cold bubbling and more than 99% of combined sulphite was recovered by heating afterwards for 10 min.The scope of the modified Rankine method was also extended to the determination of sulphite in concentrated orange juice.

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Verwendung der modifizierten Rankine-Methode zur getrennten Bestimmung von Sulfiten in Lebensmitteln. III

Zusammenfassung Die Rankine-Methode wurde bedeutend verbessert. Ein Umtausch der Aspiration mit Blasensystem trug beträchtlich zur Vereinfachung des Bestimmungsverfahrens bei, und die Reproduzierbarkeit wurde verbessert. Luft (Fließrate 1,01/min) wurde als Blasengas benutzt, da der Gebrauch von Inertgas für die Wiederfindungsrate unbedeutend ist.Natriumhydrogensulfit (freies Sulfit) und drei Arten gebundener Sulfite (Acetaldehydhydrogensulfit, Pyruvathydrogensulfit undd-Mannosehydrogensulfit) dienten dazu, die geeignetsten Bedingungen für die getrennte Bestimmung der freien und gebundenen Sulfite zu ermitteln.Freies Sulfit wurde bei 0 °C durch 30 min Durchblasen vertrieben. Hierbei ging kein gebundenes Sulîit verloren. Die Phosphorsäurekonzentration war wichtig für die Freisetzung des Sulfites. Wenn man 25%ige Phosphorsäure verwendet, werden > 99% freien Sulfites beim Durchblasen in der Kälte vertrieben, während > 99% gebundenen Sulfites durch nachheriges 10 min langes Erhitzen wiedergewonnen werden.Die modifizierte Rankine-Methode wurde weiterhin für konzentrierte Säfte verwendet.

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Studies on the Analyses of Sulphites in Foods (III)     

Depolymerization of fluoroalkyl methacrylate polymers as studied with thermogravimetry

Thermal behaviors of a few kinds of poly(fluoroalkyl methacrylate) prepared by γ-or UV-ray polymerization were investigated by using thermogravimetric measurements with the intermittent analysis of the gaseous products. The degradation of fluoroalkyl methacrylate polymers, monomeric units of which were CH₂=C(CH₃)COOCH₂(CF₂CF₂)_nH, n = 1, 2, and 3, proceeded according to the depolymerization mechanism reproducing the pristine monomer exclusively, but the thermogram in inert atmosphere showed the features of a two-step reaction. Two species of polymer differing in the heat stability were supposed to exist in the polymeric substance produced by γ- or UV-ray irradiation, and the fraction of polymer having lower heat stability increased with the increasing length of the fluoroalkyl ester group. In air, however, the thermogram of poly(fluoroalkyl methacrylate) showed no such a stepwise weight decrease as was observed in inert atomsphere with the elevating temperature, and the temperatures at which the depolymerization was introduced shifted to a much higher region. The results were ascribed to the reaction of initiating polymer radicals produced on polymer having lower stability with oxygen to form hydroperoxide, which once stabilized the polymer radicals and obstructed the initiaition of the unzipping reaction till higher temperature.     

Sensory evaluation of beef patties and sausages containing lipids with various component fatty acids

Sensory scores of beef patties and emulsion-type sausages containing varying levels of lipids with various component fatty acids were investigated. Sensory scores of beef patties containing solid fat were relatively constant at every fat level examined. However, scores of beef patties containing liquid oil decreased with increasing level of added oil. In the case of sausages, on the other hand, sensory evaluation scores markedly decreased with increasing the level of solid fat, while no significant change was observed in panel scores when liquid oil was mixed into sausages. Even 10% level of added lipids satisfactorily gave preferable acceptability to beef patties and sausages.