Search

Lactate dehydrogenase (LDH) in blood is measured using the Japanese Society of Clinical Chemistry (JSCC) method in Japan and the International Federation of Clinical Chemistry (IFCC) method in other countries. In human clinical practice, the IFCC method replaced the JSCC method due to international standardization. Moreover, veterinary LDH measurement will also eventually shift to the IFCC method. However, the relationship between the IFCC and JSCC methods for LDH in various animals and whether they can be equated or not have not yet been investigated. This study aimed to present the changes in measurements in canines and felines after switching to the IFCC method. The plasma LDH activity of canines (N=177) and felines (N=115), who visited a secondary care veterinary clinic, was measured using the JSCC and IFCC methods. The IFCC/JSCC ratio was <1.0 in 85% of canines and 88% of felines, indicating that the IFCC method tended to show lower values than the JSCC method, presumably because LDH5 is dominant among the LDH isozymes in canines and felines. The increase in the systematic errors of both assays was in the high value range, with some samples exceeding the error tolerance from near the upper end of the reference range. When switching to the IFCC method for LDH measurement in canines and felines, each institution should consider whether the reference range and clinical diagnostic values established by the JSCC method are appropriate for continued use. [ABSTRACT FROM AUTHOR]

Much attention has been focused on the moss family Mniaceae over the past 50 years, yet new discoveries continue to be made. Plagiomnium decursivum joins P. floridanum as the second new species of Plagiomnium Section Plagiomnium described in the past ten years. Careful morphological and anatomical studies, coupled with evidence from isozymes and gene sequencing, reveal that a species from Japan referred to in an earlier paper (Wyatt & Odrzykoski 1998) as "P. acutum" is actually new to science. In addition, the taxon referred to in that paper as "P. acutum-J" is in fact P. acutum, and the taxon referred to as "P. cuspidatum-F" is now named P. floridanum (Wyatt & Odrzykoski 2012). The new species, P. decursivum, is compared to its closest relative, P. acutum, and to the other four species of Section Plagiomnium. It differs in having smaller leaves with long, broad decurrencies, longer leaf cells with corner thickenings, and shorter teeth composed of a single cell. Based on isozyme data, it is concluded that allopolyploid P. cuspidatum has P. decursivum as one haploid progenitor and that P. floridanum is most likely the other. Moreover, sequencing of two regions of chloroplast DNA (rps4 and rpL16) suggests that P. floridanum may have been the maternal parent in crosses that launched the allopolyploid. The relative isolation of P. drummondii and P. japonicum within the section is apparent from both morphology and genetics. [ABSTRACT FROM AUTHOR]

Objective: We examined the change in circulating sclerostin level during the menopausal transition and we investigated the associations of sclerositin with hormones and bone turnover markers according to each menopausal stage in cross-sectional and longitudinal studies. Methods: We conducted a cross-sectional study in 200 healthy Japanese women and divided them into 4 stages (reproductive, menopausal transition, early postmenopause and late postmenopause) by menstrual regularity, follicle-stimulating hormone level and years since menopause. Serum levels of sclerostin, bone turnover markers and reproductive hormones were measured. In addition, we examined changes in sclerostin level from the reproductive stage to menopausal transition and from menopausal transition to early postmenopause in a longitudinal study. Results: In the cross-sectional study, sclerostin level gradually increased with progression of menopausal stages and showed a significant change during the menopausal transition. Sclerostin levels significantly increased from the reproductive stage to menopausal transition and from menopausal transition to early postmenopause in the longitudinal study. A negative correlation of sclerostin with estradiol was found in early postmenopause. Sclerostin levels were negatively correlated with bone-specific alkaline phosphatase levels in the reproductive stage and menopausal transition and with tartrate-resistant acid phosphatase-5b in menopausal transition. Conclusion: The change in sclerostin has already occurred in the early stage of menopausal transition and sclerostin level increases with progression of menopausal stages. Elevated sclerostin levels during the menopausal transition may be involved in relative decline in bone formation against increase in bone resorption. [ABSTRACT FROM AUTHOR]

Protobothrops tokarensis ( Pt ), a Crotalinae snake, inhabits only Takarajima and Kodakarajima islands of the Tokara Islands located in the immediate north of Amami-Oshima island of Japan. Kodakarajima P . tokarensis venom gland cDNA library gave four types of phospholipase A 2 (PLA 2 ) cDNAs encoding neutral [Asp 49 ]PLA 2 , basic [Asp 49 ]PLA 2 , highly basic [Asp 49 ]PLA 2 , and [Lys 49 ]PLA 2 . As the amino acid sequences encoded by their open reading frames (ORFs) were identical to those of PLA2, PLA-B, PLA-N, and BPI (a [Lys 49 ]PLA 2 ), respectively, from Amami-Oshima P. flavoviridis ( Pf ) venom, they were named Pt PLA2, Pt PLA-B, Pt PLA-N, and Pt BPI. Chromatography of P . tokarensis venom gave three PLA 2 isozymes, Pt PLA2, Pt PLA-B, and Pt BPI. However, BPII and BPIII ([Lys 49 ]PLA 2 s) expressed in Amami-Oshima P . flavoviridis venom were not found in P . tokarensis venom. Genomic polymerase chain reaction (PCR) for P . tokarensis liver DNAs with the unique primers gave Pt BPI gene. Notably it was found that LINE (long interspersed nuclear element)-1 fragment is inserted into second intron of Pt BPI gene. The LINE-1 fragment may prevent duplication of Pt BPI gene and thus formation of plural [Lys 49 ]PLA 2 genes in P . tokarensis genome. The interisland variegation of venom [Lys 49 ]PLA 2 isozyme genes in Protobothrops genus snakes in the southwestern islands of Japan is discussed. [ABSTRACT FROM AUTHOR]

Objective. Bokusoku (BK) is an extract from the Quercus cortex used in folk medicine for treatment of skin disorders and convergence, and is present in jumihaidokuto, a traditional Japanese medicine that is prescribed for purulent skin diseases like acne vulgaris. The excess of sebum production induced by androgen is involved in the development of acne. Our aim is to examine whether BK and its constituents inhibit testosterone metabolism and testosterone-induced sebum synthesis. Methods. Measurements of 5α-reductase activity and lipogenesis were performed using rat liver microsomes and hamster sebocytes, respectively. Results. BK dose-dependently reduced the conversion of testosterone to a more active androgen, dihydrotestosterone in a 5α-reductase enzymatic reaction. Twenty polyphenols in BK categorized as gallotannin, ellagitannin, and flavonoid were identified by LC-MS/MS. Nine polyphenols with gallate group, tetragalloyl glucose, pentagalloyl glucose, eugeniin, 1-desgalloyl eugeniin, casuarinin, castalagin, stenophyllanin C, (-)-epicatechin gallate, and (-)-epigallocatechin gallate, inhibited testosterone metabolism. In particular, pentagalloyl glucose showed the strongest activity. BK and pentagalloyl glucose suppressed testosterone-induced lipogenesis, whereas they weakly inhibited the lipogenic action of insulin. Conclusions. BK inhibited androgen-related pathogenesis of acne, testosterone conversion, and sebum synthesis, partially through 5α-reductase inhibition, and has potential to be a useful agent in the therapeutic strategy of acne. [ABSTRACT FROM AUTHOR]

Aim To provide insights into genetic differentiation between insular endemic Weigela coraeensis var. fragrans and its progenitor variety W. coraeensis var. coraeensis, the population genetic structure of both varieties was examined, and factors promoting genetic differentiation between the two taxa were explored. Location The natural range of W. coraeensis ( sensu lato) throughout mainland Japan (Honshu) and the Izu Islands. Methods The analysis included 349 and 504 individuals across the mainland (Honshu) and the Izu Islands, respectively, using 10 allozyme and 10 microsatellite loci. The population genetic structure of W. coraeensis was assessed by analysing genetic diversity indices for each population, genetic differentiation among populations, model-based Bayesian clustering or distance-based clustering, and bottleneck tests. Results The level of genetic diversity in each of the populations on the Izu Islands was negatively correlated with geographical distance between each island and the mainland. The populations on the mainland and on the Izu Islands were genetically differentiated to a certain extent; however, the microsatellite analyses suggested that gene flow also occurred between the mainland and the islands, and among individual islands. These microsatellite analyses also suggested recent bottlenecks in several populations in both areas. Main conclusions The decrease in genetic diversity throughout the Izu Islands, which correlated with distance to the mainland, Honshu, may be the result of a repeated founder effect occurring at a series of inter-island colonizations from north to south. The stepping stone-like configuration of the islands may have played a role in the dispersal of the species. Geographical isolation by sea would effectively result in genetic differentiation of W. coraeensis between mainland Honshu and the Izu Islands, although some gene flow may still occur between Honshu and the northern Izu Islands. The differentiation process of the endemic plants on the Izu Islands is anagenetic but not completed, and the study of these plants will provide insightful knowledge concerning the evolution of insular endemics. [ABSTRACT FROM AUTHOR]

Populations of Enchytraeus that reproduce by fragmentation are distributed worldwide, but their specieslevel taxonomy is unresolved because morphological differences are inconclusive. Th erefore, we compared the isozyme and RAPD-PCR patterns of 5 populations of fragmenting enchytraeids from widely distant localities (Japan, Iran, Greece, and Brazil). Among these populations, 3 were identifi ed (E. japonensis, E. bigeminus, and E. dudichi) and 2 were unidentified. Multiple isozyme bands suggested polyploidy in all investigated populations. In all RAPD-PCR algorithms except 1, the fragmenting group formed a cluster separate from 2 nonfragmenting Enchytraeus species. All 5 populations were clearly separable on the levels of protein and DNA, but the unidentifi ed Greek populations clustered more closely with E. japonensis. It remains unknown whether the fragmenting group in Enchytraeus is monophyletic or polyphyletic. Although every investigated population may deserve species rank, a consistent classification is impossible at present. [ABSTRACT FROM AUTHOR]

Serum alkaline phosphatase (ALP) activity is frequently measured in toxicity studies. Itoh et al. (2002) reported that a commercially available polyacrylamide-gel (PAG) disk electrophoresis kit used in humans (AlkPhor System, Jokoh Co., Ltd., Tokyo, Japan) for identifying serum ALP isoenzymes was useful for veterinary clinicopathological diagnosis in mongrel dogs. In the present study, based on the report of Itoh et al. (2002), we tried to expand the application range of this kit to laboratory beagle dogs which are commonly used in toxicity studies. In order to identify the origin of each ALP isoenzyme, tissue ALP extracts from the liver, bone and small intestine and serum samples were treated with neuraminidase, anti-small intestinal ALP antibody, ALP inhibitor levamisole and/or wheat germ agglutinin (WGA). The main serum ALP isoenzymes in 5-month-old intact beagle dogs were bone-derived (bone and atypical ALP: corresponding to human variant bone ALP) and they tended to decrease with age. However, liver-derived ALP isoenzyme greatly increased in the serum of cholestasis model dogs. The cholestasis model dogs also had a large molecular ALP detected in the resolving gel. This ALP could be originated from intestinal ALP or corticosteroid-induced ALP (CALP), because the activity remained even after levamisole inhibition. CALP was observed in intact laboratory beagle dogs with individual differences. These results suggest that the present method is a useful tool for detecting serum ALP isoenzymes in laboratory beagle dogs and concomitant levamisole inhibition with another gel is applicable for the evaluation of organ toxicity. [ABSTRACT FROM AUTHOR]

Weedy rice (Oryza sativa L.) plants found in Okayama, Japan, exhibit various morphophysiological characteristics, and are roughly classified into japonica-type and indica-type. We characterized the weedy strains collected from nine sites with molecular markers in flanking regions of two diagnostic isozyme loci, Acp1 and Sdh1. Haplotype analysis for closely-linked SSR markers flanking Acp1 revealed that 1) japonica-type weedy strains shared the same haplotypes with local varieties, and 2) indica-type weedy strains carrying an indica-specific isozyme allele possessed two haplotypes. One of the two haplotypes was common to a forage variety, 'Moretsu'. PCR analysis revealed that the dwarf weedy strains carried the deletion inside OsGA20ox2 as the semi-dwarf allele (sd1). The deletion was also found in 'Moretsu'. The other haplotype was shared with indica forage variety, 'Tetep', which showed the same sequence haplotype for Sdh1 with the indica-type weedy strains. The results obtained from this study suggested that the weedy strains in Okayama have multiple origins and semi-dwarfness possibly derived from 'Moretsu' or indica modern varieties. [ABSTRACT FROM AUTHOR]

We surveyed allozyme variations among ten Nymphoides coreana populations in southwestern Japan. The percentage of polymorphic loci, the mean number of alleles per locus, and the effective number of alleles per locus varied little among populations. The observed heterozygosity and expected heterozygosity were low. The proportion of gene diversity of the species within populations ( Hs) was also low. These indices all indicate that genetic diversity was low within the populations. The inbreeding coefficient within a population, Fis, was significantly larger than expected in each local population. A comparison between Hs and the proportional gene diversity of the species among populations ( Dst) showed that almost all variations occurred among populations. Large genetic differentiation was also supported by the high value of gene diversity between populations relative to the combined populations ( Gst). Nymphoides coreana is considered to be a selfing species, and genetic variations among the populations were shown to be distinct. [ABSTRACT FROM AUTHOR]

Abstract: Three species of hoplonemerteans, all very similar in external features (Oerstedia oculata, Oerstedia phoresiae, and Oerstedia zebra) were compared using 23 allozyme loci. The results of this study confirm the specific validity of O. phoresiae. At 20 of the 23 loci this species shares no common alleles with O. oculata (D =2.718) and at 21 of the 23 loci O. phoresiae shares no common alleles with O. zebra (D =2.730). Most likely O. zebra is a color variation of O. oculata, because these species are very similar in their allozyme markers (D =0.000). [Copyright &y& Elsevier]

Populations of the genus Gekko from three southern islands of the Tokara Group, Ryukyu Archipelago are known to be unique in lacking distinct preanal pores even in adult males. Recently, we found an almost identical morphotype from five islands of the Amami Group, where only G. hokouensis, a species having distinct preanal pores in adult males, has hitherto been recorded. To clarify the taxonomic status of these poreless geckos from the Tokara and Amami Groups, we conducted allozyme and morphological comparisons of Gekko specimens collected from these eight islands and one adjacent island. Results of the allozyme analyses showed the presence of fixed allelic differences at 7-14 loci between samples of the poreless form and sympatric G. hokouensis from each of the Amami islands, confirming their reproductive isolation. Our data further revealed the presence of two genetically divergent groups within the poreless form that are distinct from each other by fixed allelic differences at no less than 30% of loci examined. Ranges of these two groups are contiguous with each other but do not overlap on a single island. Distance analysis showed that one of the two groups is genetically much closer to G. hokouensis than to the other. These results strongly suggest that each of them represents an evolutionarily independent unit and deserves recognition as a full species. We describe these species based on a set of characters that clearly discriminate them from all other congeneric species hitherto described, and we briefly discuss their historical biogeography. [ABSTRACT FROM AUTHOR]

Allozyme diversity was examined in 30 populations of the maritime perennial plant Hedyotis strigulosa var. parviflora, which is distributed from subtropical islands to the central mainland of Japan. Genetic diversity within populations tended to be larger in southern island populations than in northern mainland populations. In the southern part of the distribution, the population size is generally large and populations are distributed more continuously than in the northern area, resulting in the larger effective size of southern populations as a whole. These factors play a major role in maintaining greater genetic diversity in the southern populations. By contrast, genetic diversity in the northern populations is very low, probably resulting from bottlenecks of population establishments during recolonization from refugial area to the northern areas. Geographically close populations were located near each other in the multidimensional scaling and the phenogram based on genetic distances, suggesting that gene flow among remote populations is rather limited. The pattern of genetic diversity in H. strigulosa var. parviflora is likely caused by the distribution expansion of the species; in the last glacier era, the species was restricted to the southern area; its advance to the northern area is relatively recent. Another variety endemic only to the Daito Islands, H. strigulosa var. luxurians, has lower genetic diversity than H. strigulosa var. parviflora and has genetically diverged from H. strigulosa var. parviflora. © 2008 The Linnean Society of London, Biological Journal of the Linnean Society, 2008, 93, 679–688. [ABSTRACT FROM AUTHOR]

Twelve isolates of Japanese Phytophthora infestans, which differed from the major genotypes US-1, JP-1, JP-2, and JP-3, were analyzed for RG57 fingerprints, mtDNA haplotypes, two allozyme genotypes, and mating types. Genotypes JP-1.1, JP-2.1, JP-2.2, JP-3.1, and JP-4 were newly defined. JP-1.1 and JP-2.1 were isolated discontinuously from potato fields in several years, and JP-1.1 was found in Hokkaido and Kagoshima. These results show that some minor genotypes can overwinter and disperse from their original site. [ABSTRACT FROM AUTHOR]

Abstract: Postmortem studies regarding schizophrenia revealed altered expression of genes related to γ-amino butyric acid neurotransmission system. One of the most consistent findings is the reduced level of 67 kDa glutamic acid decarboxylase isoform (GAD67). Moreover, several studies reported positive associations between the GAD67 gene (GAD1) and schizophrenia. These reasons, motivated us to carry out replication study regarding association between GAD1 (fourteen tagging SNPs) and schizophrenia in Japanese population (562 schizophrenic patients and 470 controls). However we couldn''t confirm significant association that had been previously reported. Considering size of our sample and strategy that corresponds well with the approaches used in gene-based association analysis, our conclusion is that GAD1 does not play a major role in schizophrenia in Japanese population. [Copyright &y& Elsevier]

The loach Misgurnus anguillicaudatus is a freshwater fish which include natural polyploids and clones besides most bisexually reproducing diploids. Recent genetic studies using allozymes differentiated the populations in northern region of Hokkaido from other populations in Japan. Here, we developed 23 new sets of polymerase chain reaction primers to detect polymorphic microsatellites loci and tested them in 20 individuals from Memanbetsu town (northern Hokkaido) and Kita village (southern Hokkaido). These markers can be potentially diagnostic for identification of two populations. [ABSTRACT FROM AUTHOR]

Using starch gel electrophoresis, geographical enzyme variation was surveyed in 46 populations of the Japanese salamander Hynobius nebulosus, which occurs widely in western Japan. This species exhibits substantial local genetic differentiation and is more diverse in inland regions than on small islands. In several different analyses, two groups of populations, one from Kyushu Island to the westernmost part of Chugoku district, Honshu (western group), and another from Shikoku through Kinki to Chubu district, Honshu (eastern group), were consistently recognized. Most of the remaining populations were placed in two less clearly-defined groups: the montane group from the Chugoku Mountains and the Chugoku group from the coastal regions of Chugoku district. The divergence patterns of H. nebulosus are thought to be related to the geological history of its range. The populations near the boundary of the western group were found to share several alleles that were predominant on both sides of the boundary, indicating past secondary contact. Differentiation in the montane group did not follow the isolation by distance model, probably because several populations included were genetically mixed with lowland populations of the Chugoku group. © 2006 The Linnean Society of London, Biological Journal of the Linnean Society, 2006, 89, 311–330. [ABSTRACT FROM AUTHOR]

Abstract  The genetic population structure of the Japanese freshwater goby Gymnogobius castaneus was investigated on the basis of analysis of gene products of 19 allozyme loci. Two diverged groups were detected, one being endemic to the Kanto region and the other extensively distributed in eastern Japan. These two groups were distinguishable from each other by a complete allelic substitution in one locus, G3PDH*. In the Kanto region, both groups were distributed in the same river basin, being distinguishable by a complete allelic substitution in four loci, G3PDH*, GPI-2*, PGDH*, and PGM*. These results suggest that these two groups showed reproductive isolation. [ABSTRACT FROM AUTHOR]

The phylogeography of Japanese conifers studied using polymorphic allozyme and organelle (chloroplast and mitochondrial) DNA markers is reviewed. Three Abies species showed a clear geographical cline based on mtDNA polymorphisms. Allozyme markers revealed a clear genetic cline in A. mariesii, with no mtDNA polymorphism. Refugia of these species during the last glaciation were probably located along the southwestern Pacific side of Japan. Based on allozyme analysis, Pinus pumila may have migrated from Siberia southward from Hokkaido to higher mountains in central Japan. Mitochondrial DNA polymorphisms clearly differentiated the pentaphylla and parviflora varieties of Pinus parviflora and suggest that refugia of the latter species were located south of the central mountains of Chubu district during the last glaciation. Current populations in the Chubu highlands, Tohoku, and Hokkaido presumably became established by colonization from these refugia. Phylogeographic analysis of Cryptomeriajaponica using allozymes and DNA markers did not reveal distinct patterns. However, microsatellite analysis revealed highest allelic diversity in putative refugia of the last glaciation. Allozyme analysis of Chamaecyparis obtusa revealed a clear cline. Recently, substantial numbers of loci based on expressed sequence tags for population and genome mapping were developed. Cryptorneria japonica and Chamaecyparis obtusa belong to different families, but molecular phylogeny based on chloroplast sequence data revealed that they are closely related and that sequence tag site markers of C. japonica are highly applicable to C. obtusa. Using such markers, researchers have attempted to clarify the genetic structure of the two species. This paper also discusses the most successful approaches (to date) for elucidating the genetic structure of the above-mentioned species and other phylogeographic issues related to Japanese conifers. [ABSTRACT FROM AUTHOR]

Isozyme analyses were conducted to study the geographic variation of Capsicum frutescens L. in Southeast and East Asia, and to investigate its dispersal routes into Japan. Eight enzymes (EST, FM, G6PD, GR, ME(A), PGI, PGM, ShDH) were variable among accessions of C. frutescens in Southeast and East Asia. Accessions from the Ryukyu Islands were closely related to those in Indonesia, whereas accessions from the Bonin Islands showed exactly the same isozyme patterns as those from Indonesia and Northern Thailand. Accessions in the Ryukyu Islands were different from those in the Bonin Islands, suggesting that there may be two independent dispersal routes into Japan. One route was from Indonesia via the Philippines or Taiwan, or directly to the Ryukyu Islands, and another was from Indonesia via the Mariana Islands, or other islands in the Pacific, to the Bonin Islands. [ABSTRACT FROM AUTHOR]

Features the genetic diversity of the vulnerable aquatic macrophyte Sparganium erectum and its congeners in rural wetland in Japan. Decrease of habitats of Sparganium erectum L through the development of modern cultural practices of rice; Significance of the of the shape of inflorescence and fruit traits in an emerged condition on genus characters; Result of the comparison between genetic distances among genotypes with geographic distances among natural populations.

Genetic variation at 10 allozyme loci was analyzed in 14 populations ofPolygala reinii(Polygalaceae), a perennial herb endemic to central Honshu, Japan, with a fragmented geographical distribution. The levels of genetic variation within species (P=80.0,A=3.10,HE=0.303) and within populations (P=42.1,A=1.61,HE=0.163) were considerably higher than the mean for other endemic plants or short-lived perennial herbs. Genetic differentiation among populations was also high (GST=0.404). The genetic distance phenogram tended to show a clustering of the populations reflecting the fragmentation of the species range. A principal component analysis revealed the same tendency, as well as three groupings of populations in the Tokai district, on the Kii Peninsula and in the northern Kinki district. A negative correlation was obtained between the levels of gene flow and geographical distance among the populations (r=-0.745,P<0.0001). These results indicated limited gene flow among populations inP. reinii, presumably due to the geographical isolation accompanying the fragmented distribution. On the other hand, the geographical differentiation between the Japan Sea and Pacific Ocean sides was found inP. reinii, suggesting the influence of postglacial migration on the establishment of the genetic structure of this species. [ABSTRACT FROM AUTHOR]

Genetic diversities of the nuclear and mitochondrial genomes in Pinus parviflora were studied in 16 populations, which were distributed across most of the species' range in Japan. Six mitochondrial DNA haplotypes were identified among the 16 populations. The intrapopulation diversity of allozymes was similar to that of other endemic woody species (HS=0.259). Although P. parviflora is distributed in discrete populations, differentiation between these was very low (GST=0.044). In addition, the extent of genetic differentiation between two varieties (var. pentaphylla and var. parviflora) was extremely low (GVT=0.001). Intrapopulation diversity of mitochondrial DNA was also very low (HS=0.098), but population differentiation was high (GST=0.863). Moreover, the distribution of haplotypes reflected the taxonomic differences between P. parviflora var. pentaphylla and var. parviflora. The populations of var. pentaphylla and var. parviflora contained different haplotypes. Differing modes of inheritance may account for the differences in nuclear and mitochondrial genetic diversity.Heredity (2003) 91, 510-518. doi:10.1038/sj.hdy.6800349 [ABSTRACT FROM AUTHOR]

From 25 populations of Pseudorasbora in Japan, polymorphism at 22 allozyme loci indicated that the level of genetic differentiation between Pseudorasbora parva and Pseudorasbora pumila (D=0·421-0·517) was greater than that between the two subspecies of P. pumila, P. pumila pumila and P. pumila subsp. (D=0·164), consistent with morphological differences. While P. parva displayed genetic variation (H=0·003-0·100) similar to other freshwater fishes, P. pumila pumila and P. pumila subsp, populations showed no genetic variation. In five of the 15 populations collected from the contact zone between P. parva and P. pumila pumila, hybrids were detected by allozyme analyses. All hybrids were presumed to represent the F[sub 1] generation, because they were heterozygous at all 12 loci diagnostic between P. parva and P. pumilapumila. Although four populations were characterized by high frequencies of F[sub 1] hybrids (c. 40%), only one of the parental species was observed in each case. The results indicated that the two species have hybridized easily under natural conditions, but cannot coexist in the long-term. It is suggested that continued invasion of P. parva would hasten the extinction of P. pumila pumila. [ABSTRACT FROM AUTHOR]

The genetic variation in Eumeces latiscutatus from the main islands of Japan and the northern Ryukyus was investigated and compared with that of its close relatives (E. okadae and E. marginatus) using allozyme data. All three samples from the northern Tokara Island Group, currently identified as E. latiscutatus, were shown to belong to E. marginatus and not to E. latiscutatus. The non-monophyly of the northern Tokara samples and the great genetic differences within these samples may have resulted from colonization from more than one source population by northeastward overseas dispersal. The samples from the Izu Peninsula were genetically much closer to E. okadae than the other conspecific ones. This indicates that the samples from the Izu Peninsula and the other samples of E. latiscutatus should be treated as distinct species, and that E. latiscutatus from the Izu Peninsula and E. okadae from the Izu Island Group may be treated as conspecific. Samples from western Japan were genetically well differentiated from those of eastern Japan. Within the western group, the samples from the Osumi Island Group was genetically distinct from those from the other regions, by possessing unique alleles. Our phenograms also reveal a distant affinity between samples from the Danjo island Group and the main islands of Japan+ This may be the result of long geographic isolation of the Osumi and Danjo Island Groups from Kyushu. By contrast, samples from Sapporo and Aomori were poorly differentiated genetically in spite of the long separation of these two localities by the Tsugaru Strait. This suggests that overseas dispersal of E. latiscutatus occurred across this strait after its formation. [ABSTRACT FROM AUTHOR]

Genetic variation of Japanese rice cultivars were examined. Five of 450 lowland cultivars and another five of 200 upland cultivars were determined as the indica type by using isozyme genotypes and the remainder were of the japonica type. The major characteristics of these indica cultivars, revealed a slender shape of grains, a short apiculus hair length, a positive allele for Ph reaction, and allele-3 for the Pgd1 locus. Three of these indica cultivars showed a non-deletion ORF100, which is essential to the japonica-type plastid. The plastid subtype identity (PS-ID) sequences of these plastids is 6C7A, which is also a japonica-specific repeat unit. Thus, these cultivars were concluded to be naturally generated cytoplasm substituted lines. These plastids were introduced into a indica genetic background from japonica cultivars grown elsewhere. The rest of the indica cultivars revealed a deletion-type ORF100 and plastid subtype 8C8A, both of which are indica-specific. These cultivars carried indica-type allelic constitutions for diagnostic isozyme loci. However, other characters were identical to the cytoplasm-substituted cultivars in Japan. In East and Southeast Asia, cultivars carrying a indica-type nuclear genotype with a japonica-type plastid are restricted to Aus cultivars in the Bengal region. Genetic and historical records suggest that Japanese indica cultivars and the Aus cultivars are closely related. The Aus cultivars acquire necessary genetic constitutions from both indica and japonica cultivars through naturally occurring out-crossing to adapt to a particular cultivation condition in the region. The wide adaptability enabled them to be introduced into a northern region like Japan. [ABSTRACT FROM AUTHOR]

Characterization was investigated on the 38 kDa and 42 kDa chitinase (EC3.2.1.14) isozymes from the liver of Japanese common squid Todarodes pacificus . Optimum pH toward colloidal chitin was observed at pH 3.0 for the 38 kDa chitinase, and pH 3.0 and 9.0 for the 42 kDa chitinase. K m and k cat of the 38 kDa and 42 kDa chitinases toward a longer substrate, glycol chitin, were 0.071 mg/mL and 1.22/s, and 0.074 mg/mL and 0.196/s, respectively. Alternatively, strong substrate inhibition of both chitinases were observed toward a short substrate, N -acetylchitopentaose (GlcNAc5 ). Both chitinases decomposed not only chitin but also chitosan (D. A. 95%). The cleavage pattern and reaction rate were investigated using N -acetylchitooligosaccharides (GlcNAcn , n = 2–6). Both chitinases hydrolyzed GlcNAcn (n = 4,5, and 6). The release of GlcNAc was not observed. The speed of the reaction was observed to be in the following order: GlcNAc4 > GlcNAc5 > GlcNAc6 for the 38 kDa chitinase, and GlcNAc6 > GlcNAc5 > GlcNAc4 for the 42 kDa chitinase. Both the chitinases released p -nitrophenol from p -nitrophenyl GlcNAcn (n = 2, 3, and 4). N-terminal amino acid sequences of the 38 kDa and 42 kDa chitinases were YLLSXYFTNWSQYRPGAGKYFPQNI and EYRKVXYYTNWSQYREVPAKFFPEN, respectively. [ABSTRACT FROM AUTHOR]

The influence of fragmentation on local genetic population structure of the fluvial sculpin Cottus nozawae was studied by comparing populations in two regions: the Tohoku District, the southern margin of the species native range, and Hokkaido Island, the center of its native range, in Japan, based on restriction fragment length polymorphism analysis of mitochondrial DNA amplified by means of the polymerase chain reaction. Relatively high genetic divergences were found among populations from different regions. Differences in the mobility of organisms and their distributional fragmentation determine their phylogeographic structure. Various population genetic models such as isolation by distance, and stepping-stone models have been proposed for estimating the effect of fragmentation on the distribution of genetic variation. However, empirical study of the spatial genetic structure of fragmented populations of fishes is rare. Genetic divergence between the populations from Hokkaido Island and the Tohoku District was high, based on allozyme variation and restriction fragment length polymorphism analysis of whole mitochondrial DNA. The present study tested the hypothesis that if levels of population subdivision are affected by the distributional pattern, genetic divergence among populations within river systems in the distribution area at the southern margin, Tohoku District, would be higher than that in the north-central area, Hokkaido Island. Genetic variation was estimated by grouping the studied populations into the two main regions and separately analyzing populations among river systems within a single region and among tributary populations within a single river system.

Examines the mating system and population genetics of an androdioecious tree Fraxinus lanuginosa Koids in Hokkaido, Japan. Use of electrophoretic allozyme methods; Variations in male frequency; Estimation of outcrossing rates.

Gene products of 18 allozyme loci from 1268 individuals of a Japanese freshwater goby called donko, Odontobutis obscura (Odontobutidae; Gobioidei), from 33 localities in the Koya River, Yamaguchi Prefecture, Japan, were investigated to determine the extent of genetic divergence and gene flow within a river metapopulation. Genetic indices including GST(mean FST 0.182), FIT(mean 0.192) and D(mean 0.015) indicated a considerable divergence of local populations in the river. The genetic distance (D) and channel distance between pairs of populations did not show a good correlation, and geographical neighbors were not always genetic neighbors. Therefore, the genetic divergence of populations is attributable to independent genetic drift with restricted gene flow among populations. The agricultural dams and weirs constructed across the river must be responsible for the restricted gene flow. The metapopulation structure of O. obscura in the Koya River may be barely sustained by one-way gene flow only from the upper to the lower populations. An occasional artificial transplantation of some individuals from the lower to the upper populations may be one alternative to maintain a river metapopulation structure safely. [ABSTRACT FROM AUTHOR]

Objective: Although most patients with type 1 diabetes are considered to have T-cell-mediated autoimmune disease, a method of measuring of pancreatic beta-cell-specific T-cell function in cases of type 1 diabetes has yet to be established. Here, we focused on interferon-inducible protein-10 (IP-10), a chemokine that promotes the migration of activated T-helper 1 (Th1) cells and measured serum IP-10 levels in patients with human type 1 diabetes, which is regarded as a Th1-mediated disease.Research Design and Methods: Serum samples were obtained from diabetic patients, and the levels of autoantibodies (GAD and insulinoma-associated protein-2 [IA-2]) and IP-10 were measured. Diabetic patients positive for either or both of the autoantibodies were classified as Ab+ type 1, and those negative for both were classified as Ab type 1. To evaluate islet antigen-specific responses, peripheral blood from patients stimulated with or without GAD was used, and intracellular cytokine staining for flowcytometry was performed.Results: The Ab+ and Ab- type 1 groups both showed a significantly higher serum IP-10 level than the healthy subjects (P < 0.001 and P < 0.05, respectively), and the IP-10 level in the recent-onset Ab+ subgroup was significantly higher than that in the established (longstanding) Ab+ subgroup (P < 0.002). Furthermore, there was a significant positive correlation between the serum IP-10 level and the number of GAD-reactive gamma-interferon-producing CD4+ cells in the Ab+ type 1 group (P < 0.007).Conclusions: Our findings demonstrate that measurement of serum IP-10 concentrations is useful in patients with type 1 diabetes. [ABSTRACT FROM AUTHOR]

SUMMARY:Allozyme variation of diploid loach Misgurnus anguillicaudatus was examined in samples collected from 44 localities in Japan. Genetic differentiation was estimated based on allele frequencies of 12 enzyme loci. Nine loci (36.9%) were polymorphic (< 0.95 in major allele frequency) and average heterozygosities were unusually high (Ho = 0.110 ± 0.051, He = 0.131 ± 0.052). The coefficient of gene differentiation (Gst = 0.774) suggested high genetic differentiation among populations. Based on the unweighted pair-group method using arithmetic averages (UPGMA) dendrogram, loach populations were grouped into six clusters. The population of Memanbetsu, Hokkaido (group 1) was remarkably different from others (D = 0.286). Populations distributed in Niigata, Nagano, Tochigi, Saitama, Chiba and Shizuoka Prefectures (group 6) were also genetically distinct from other loaches (D = 0.192). These results suggest genetic differentiation among groups 1, 6, and other loaches may be inter-subspecies level. Other loaches could be separated to four clusters (groups 2–5) at D = 0.100–0.065. Genetic differentiation among them seems to be at local race level. [ABSTRACT FROM AUTHOR]

Genetic variability was examined at 16 putative allozymic loci in the narrow endemic fern Polystichum otomasui. Although this species is distributed in only a few valleys within an approximately 10 km×6 km area on the Kyusyu Island of Japan, there are a relatively large number of individuals in this area. The percentage of polymorphic loci (P), the number of alleles per locus (A), the observed heterozygosity (HO) and the expected heterozygosity (HE) were 61.9, 1.93, 0.166 and 0.177, respectively, at the population level, and P, A and HE were 81.3, 2.69 and 0.199, respectively, at the species level, indicating that P. otomasui has extremely high allozymic variation for a fern species. The GST value among the 10 populations examined within the range was 0.108, suggesting that the interpopulation gene flow is high enough to impede genetic structuring. The mating system estimates, Wright's fixation index and the intragametophytic selfing rate, indicate that the species is outcrossing. High genetic variability, in spite of narrowness of the distribution, may have resulted from a recent rapid decline in the population and/or the large effective population size resulting from the extensive gene flow among the populations. [ABSTRACT FROM AUTHOR]

The effects of introgression between two sympatric species on intraspecific genetic variation were studied in the land snail species Mandarina anijimana and M. mandarina on the island of Anijima. Allozyme analysis suggests that extensive introgression between this sympatric pair of species is occurring at several sites. In spite of this extensive gene flow, these species remain morphologically distinct. This suggests that distinctive sympatric forms can persist for long periods despite significant gene flow between them. Because there are no effective geographical barriers in the island, the geographical variation within species in allozyme frequencies shown here are mostly produced by geographical variation in the level of gene exchange between the two species. These breakdowns in reproductive isolation between species mostly occur in populations in the transitional zone between shrub and forest or at sites with poor vegetation. Mandarina anijimana is rare in the transitional zone and is never found in the forest, but alleles introduced from M. anijimana are found at high frequency in the populations of M. mandarina in these areas. This suggests ‘frequency-dependent’ hybridization in which the rare species at a site mates with individuals of the other sympatric species. [ABSTRACT FROM AUTHOR]

Current-year needles from individual trees were used to study the genetic diversity in 10 natural populations of Picea glehnii (Masters) in Hokkaido, Japan, by polyacrylamide gel electrophoresis. Data from 12 polymorphic loci encoding 33 alleles identified by nine enzyme systems were analysed. Mean number of alleles per locus, percentage polymorphic loci and observed and expected heterozygosities were 1.98, 75 per cent, 0.08 and 0.088, respectively. These genetic parameters varied considerably among populations. Diversity among the populations was small with a mean F[sub ST] of 0.022 and genetic distance of 0.0017. However, a X²-test showed that allele frequencies were different (P < 0.05) among the populations at 10 of the 12 loci; cluster and canonical discriminant analyses indicated that some of the populations were very different from others; and correlation analyses revealed significant relationships between some of the allele frequencies and longitude, latitude and altitude. Results suggest that genetic variation in P. glehnii is both geographically clinal and population-specific. [ABSTRACT FROM AUTHOR]

Isoenzyme variation of 198 isolates of Phytophthora infestans collected from many locations in Japan during 1987-90, and of four pre-1987 isolates, was examined using starch gel electrophoresis. A previously unreported allele at malic isoenzyme locus/ME (90) was observed. An association between mating types and isoenzyme genotypes at three isoenzyme loci, glucosephosphate isomerase (GPI-1), peptidase (PEP-l) and ME, was found. At PEP-1, the A2 isolates from Japan had a previously unreported genotype (96/96). Normal segregation at the malic isoenzyme locus occurred in a cross of Japanese and Mexican parents. The results provide evidence of a change in the population genetic structure of P. infestans in Japan. [ABSTRACT FROM AUTHOR]

The demographic genetic structure of a metapopulation of the Japanese beech (Fagus crenata Blume) in the Ogawa Forest Preserve was analyzed using allozyme variabilities as genetic markers. A total of 138 mature trees in an area of 30 ha were first mapped; their DBH was measured and leaf samples were collected and genotyped. A total of 38 multilocus genotypes in a combination of six loci and 22 alleles were detected. Spatial analyses, using Moran's/and coancestry (Rij) showed there was no conspicuous pattern of associations between genetic similarities and individual distances in scales larger than 30 m. In smaller, 5 m interval scales, however, gene dispersal from mother trees to juvenile trees was found in the ranges of 10 to 20 m in distance. The indirect measures of gene flow using multilocus genotypes revealed that most progenies were derived from the nearest patch populations and also from neighboring patch populations. Genetic subdivisions of the Ogawa metapopulation were analyzed on the basis of patch populations (Gst=0.234) and local populations (Gst=0.094), reflecting the effects of population fragmentation. Genetic neighborhood area (A) obtained in Ogawa was 3050-4091 m², and effective population size (Ne) was 1.2-2.1 mature individuals. A very small Ne in Ogawa may reflect topographic complexity and past artificial disturbances of the habitats. [ABSTRACT FROM AUTHOR]

We investigated chronological changes in allozyme diversities in Fagus crenata populations by means of Size-class discriminations, using allozyme variations in two transects established in a mixed beech-oak forest in the Ogawa Forest Preserve. All individuals within transects were divided into six size-classes based on their diameters: class 0, seedlings; class 1. juveniles <5 mm; class 2, 5 < juveniles < 10 mm; class 3, 10 < juveniles < 25 mm; class 4, 25 Adh, Pgi and Pgm2 (in Transect 1) and Got2 (in Transect 2). The differences in genetic structures among different size-classes showed the effects of past repeated reproductive events by different mother trees, some of which had already died. Examination in the year that they had become established (1994) and three years (1997) later showed a considerable decline in the number of multilocus genotypes. Thus, the current state of genetic diversity among all individuals in the Ogawa population that we sampled reflects the consequences of repeated reproductive events and complex interactions among various biotic elements and abiotic environmental regimes in the habitats. [ABSTRACT FROM AUTHOR]

Ainsliaea apiculata is a cleistogamous perennial. This species occasionally hybridizes with A. faurieana on Yakushima Island, Japan and the hybrids are fertile. A cytological study of these hybrids revealed no irregularity in their male meiosis and the occurrence of genetic segregation through successful reproduction. Allozyme variation provided evidence for segregation through selfing in the hybrids. Observation on pollen grain number showed that these hybrids have cleistogamous flowers in various frequencies. Cleistogamy is dominantly inherited in the hybrids and selfing by cleistogamous flowers is considered to contribute to the formation of hybrid populations. [ABSTRACT FROM AUTHOR]

The article examines the study concerning differentiations of allele frequency at Malic enzyme-1 locus among subspecies of white-spotted char, Salvelinus leucomaenis, in Japan. In the study, the researchers gathered char samples from different regions of the country. Allozyme analysis and genetic differentiation were done within and between local samples. Furthermore, the result indicated that a allele was dominantly observed in 14 samples from Lake Shikotsu to Hino River. On the other hand, a larger range of diversified allele frequency was found in eight samples from the Hakuta River to Takatsu River.

Describes the purification and properties of chitinase isozyme from the liver of Japanese common squid Todarodes pacificus. Purification steps; Hydrolization of chitin to N-acetylchitooligosaccharides; Chitinase activity; Hydrolysis of chitosan.

In what has become known as the Yusho incident, thousands of people in western Japan were poisoned by the accidental ingestion of rice bran oil contaminated with polychlorinated biphenyls (PCBs) and various dioxins and dioxin-like compounds. In this study, we investigated the accumulation patterns of 69 PCB congeners in the blood of Yusho patients in comparison with those of non-exposed controls. The blood samples were collected at medical check-ups in 2004 and 2005. To compare the patterns of PCB congeners, we calculated the concentration ratio of each congener relative to the 2,2',4,4',5,5'-hexaCB (CB153) concentration. The concentration ratios of tetra- and penta-chlorinated congeners in the blood of Yusho patients were significantly lower than those of controls. To examine the cytochrome P450 (CYP)-dependent metabolic potential of the 2,3',4,4'5-pentaCB (CB118), CB153, and 2,3,3',4,4'5-hexaCB (CB156) congeners, we conducted PCB-CYP (CYP1A1, CYP1A2, CYP2A6, and CYP2B6) docking simulation by in silico analysis. The docking models showed that human CYP1A1, CYP2A6, and CYP2B6 isozymes have the potential to metabolize CB118 and CB153. On the other hand, it was inferred that CB156 is difficult to be metabolized by these four CYP isozymes. These results indicate that CYP1 and CYP2 isozymes may be involved in the characteristic accumulation patterns of PCB congeners in the blood of Yusho patients.

Key Message: Suaeda maritima varieties native to Japan and Egypt were cultured under aseptic conditions. The varieties differed in genetic distance but exhibited similar expression profiles of superoxide dismutase isozyme genes. The expression characteristics of superoxide dismutase (SOD; EC 1.15.1.1) isozyme genes from halophytic Suaeda marit ima plants native to Japan and Egypt were analyzed using young plants grown under aseptic conditions. A phylogenetic tree based on internal transcribed spacer sequences suggested that Egyptian S. maritima is related to European and India S. maritima, while Japanese S. maritima belongs to a separate clade. An in-gel SOD activity staining assay revealed that leaves from both the Egyptian and Japanese varieties showed high levels of CuZn-SOD and Fe-SOD activity, but no Mn-SOD activity; conversely, stems from both varieties showed Mn-SOD activity as well as other SOD isozyme activities. In Japanese S. maritima leaves, SOD activity was increased by incubation in growth medium containing 400 mM NaCl, while Egyptian S. maritima leaves showed elevated SOD activity in the absence of high salt. Genes encoding Mn-SOD and Fe-SOD were isolated from both plant types. RT-PCR analysis revealed that all SOD isozyme-encoding genes were expressed at the same levels in leaves from both plant types grown in normal or high-salt medium. In contrast, the expression of genes encoding choline monooxygenase and betaine aldehyde dehydrogenase, which are involved in betacyanin biosynthesis, was increased in high-salt medium. In leaves of Japanese S. maritima plants, Fe deficiency without high salt exposure preferentially decreased Fe-SOD activity. On the other hand, Fe deficiency with high salt exposure decreased not only Fe-SOD activity but also CuZn-SOD activity, suggesting that Fe availability is involved in the up-regulation of SOD isozymes mediating salt tolerance.

Phylogenetic relationships of the Chugoku-Kyushu and Shikoku groups of the Japanese spinous loach, Cobitis takatsuensis, among the diploid congeners in Japan, C. biwae and Cobitis sp. complex with Niwaella delicata as an outgroup, were investigated by analyzing 20 protein-coding loci. The two groups of C. takatsuensis are clearly diverged genetically, with the genetic distance corresponding to the species level (average D?=?0.27). The two groups form a monophyletic cluster (bootstrap probability, P?=?94.9%) that is a sister cluster of C. biwae ( P?=?63.1%). The monophyletic cluster of the Cobitis sp. complex ( P?=?93.2%) is the most distantly related in the genus. The present results differ mostly from the mitochondrial phylogeny previously known in which the Chugoku-Kyushu group of C. takatsuensis is connected to the eastern group of C. biwae and the Shikoku group is connected to the Cobitis sp. complex with the western group of C. biwae. The contradiction between the allozymic and mitochondrial phylogenies suggests that the Shikoku group of C. takatsuensis and the western group of C. biwae received mitochondrial introgression from the Cobitis sp. complex. [ABSTRACT FROM AUTHOR]

Leigh syndrome (LS) is an early-onset progressive neurodegenerative disorder characterized by unique, bilateral neuropathological findings in brainstem, basal ganglia, cerebellum and spinal cord. LS is genetically heterogeneous, with the majority of the causative genes affecting mitochondrial malfunction, and many cases still remain unsolved. Here, we report male sibs affected with LS showing ketonemia, but no marked elevation of lactate and pyruvate. To identify their genetic cause, we performed whole exome sequencing. Candidate variants were narrowed down based on autosomal recessive and X-linked recessive models. Only one hemizygous missense mutation (c.665G>C, p.W222S) in glycogenin-2 (GYG2) (isoform a: NM&#95;001079855) in both affected sibs and a heterozygous change in their mother were identified, being consistent with the X-linked recessive trait. GYG2 encodes glycogenin-2 (GYG2) protein, which plays an important role in the initiation of glycogen synthesis. Based on the structural modeling, the mutation can destabilize the structure and result in protein malfunctioning. Furthermore, in vitro experiments showed mutant GYG2 was unable to undergo the self-glucosylation, which is observed in wild-type GYG2. This is the first report of GYG2 mutation in human, implying a possible link between GYG2 abnormality and LS.

Objective: We investigated the association between endometriosis and polymorphisms in the N-acetyltransferase 1 (NAT1) and N-acetyltransferase 2 (NAT2) genes in a Japanese population, having previously demonstrated a positive association with NAT2 polymorphisms in a UK population., Methods: Genotyping for NAT1 alleles *3, *4, *10, and *11, and NAT2 alleles *4, *5A, *5B, *5C, *6A, and *7B was performed using polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) and allele-specific PCR (AS-PCR) analysis in 145 ethnically Japanese, endometriosis patients and 182 controls. The NAT1 and NAT2 allele and genotype frequencies were compared in cases and controls using the Fisher exact test., Results: No significant differences between cases and controls were observed in the frequencies of the NAT1 and NAT2 alleles (P = .13; P = .91) and genotypes (P = .24; P = .79), and the NAT2 acetylation phenotypes (P = .46). Dividing the cases into a subgroup, consisting of women with severe disease only (n = 80), had no effect on the results., Conclusion: The distribution of NAT1 and NAT2 allele and genotype frequencies were not significantly different between Japanese cases and controls. Our findings suggest that polymorphisms in NAT1 and NAT2 are unlikely to be associated with an increased risk of endometriosis in the Japanese population.

Broadcast-spawning corals expel eggs and sperm, and the fertilized eggs develop into planulae in the water column. As these sessile corals generally disperse during the planktonic larval stage, their larval characteristics (e.g., survival and settlement rates) are thought to be important for their dispersal. Although some studies of coral larval dispersal have focused on the maximum time that larvae can remain viable and settle, the relevance of this maximum settlement competency period for long distance dispersal remains unclear. To examine the relationship between competency periods and genetic differentiation, we performed laboratory experiments to investigate settlement rates of planulae and determine the degree of genetic differentiation in Acropora digitifera in the Ryukyu Archipelago, southern Japan. In addition, we compared our findings to published data for A. tenuis, which was studied using our methods. Our results indicated that the maximum settlement competency period was lower in A. digitifera planulae (54 days) than in A. tenuis (69 days) planulae. The mean survival rates at 45 days and 59 days after spawning were less than 10%. Furthermore, percentages of planulae that remained viable and settle at 30 days after spawning (survival rate x settlement rates at 30 days) were approximately 18% and 25% in A. digitifera and A. tenuis, respectively. By contrast, gene flow (N(e)m: number of migrations per generation) was significantly higher in A. digitifera (7.8 to 41.4) than in A. tenuis (3.1 to 22.5). These results indicate that the settlement competency period and survival rates are unlikely to be robust predictors of gene flow. Overall, we detected significant genetic differentiation between Kerama and Okinawa in A. digitifera. As direct observation of planula dispersal between Kerama and Okinawa has been reported, we concluded that genetic mixing is not complete, but that some localized planulae may disperse from Kerama to Okinawa via a specific current depending on reef or locality.

In clinical practice, one drug is frequently used in combination with one or more other drugs, rather than as a sole regimen, and therefore healthcare providers need to carefully consider drug interactions. As mechanisms of drug interactions, metabolic enzymes of drugs are seen as one of the most likely interactive sites, where a majority of drugs are metabolized by cytochrome P450 (CYP). For this reason, providing appropriate information on CYP in package inserts is of grave importance. In fact, the package insert is the primary tool for supplying information on drugs to healthcare providers. The present study was designed to determine how many package inserts of prescription drugs marketed in Japan were providing CYP information. We searched the April 2003 version of "Drugs in Japan DB," which listed 2,022 prescription drugs, and found that only 239 package inserts (11.8%) mentioned CYP information and that only 194 (9.6%) specified CYP isozymes. To assess the improvement of package inserts, we searched "Drugs in Japan DB" from the January 2000 version to the April 2003 version. We found that CYP information had increased year by year (eg, 7.8-11.8% annually). For newly approved drugs, an analysis of the relationship between approval year and CYP information in package inserts (April 2003 version) revealed that recently approved drugs had more CYP information (eg, 45.5-51.3% of drugs in 1999-2002, compared to 6.8-26.1% in 1991-1996). A search for regulatory review documents for new drugs approved from 1999 to 2002 suggested that this recent improvement could be related to the increased number of studies identifying CYP isozymes involved in the metabolism or interaction with other drugs. Another reason for the recent improvement may be the fact that the guideline for package inserts for prescription drugs was revised in 1997, and the guidelines for drug interaction and pharmacokinetic studies were published between 1997 and 1999.