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    Natural Conversation Reconstruction Tasks: The Language Classroom as a Meeting Place
    Ohashi, J (University of Technology, Sydney (UTS), 2009)
    This paper, drawing on Pratt’s notion of ‘transculturation’ and Bhabha’s ‘third space’, presents an example of language learning tasks that empower learners’ agency and promote their cross-cultural awareness and sensitivities to a different set of cultural expectations, using a naturally occurred Japanese thanking episodes. The paper discusses the merits of Natural Conversation Reconstruction Tasks (NCRTs) as a practical method for helping L2 learners develop this ‘intercultural competence’. It is based on a qualitative study of the results of one NCRT created for use in the context of teaching Japanese as a L2 in a multicultural society. It suggests the NCRT encourages the learners to explore the intersection where language use, speaker intention and L1 and L2 cultural norms meet. Such a process helps the learners become aware of socially expected patterns of communication in L1 and L2 in terms of the choices of speech act, formulaic expressions, sequential organization and politeness orientation. The learners’ comments suggest that the NCRT helps learners transcend their cultural boundaries by overcoming their narrow understanding of ‘thanking’ as ‘expressions of gratitude and appreciation’ and by cross-culturally widening their views of what counts as thanking. The NCRT with rich contextual information promotes the learners’ intercultural awareness, sensitivity to context and intercultural exploration in the space between L1 and L2, where they have authority and freedom of making sense of conversations, and pragmatics is fully integrated into language pedagogy.
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    ACE1 polymorphism and progression of SARS.
    Itoyama, S ; Keicho, N ; Quy, T ; Phi, NC ; Long, HT ; Ha, LD ; Ban, VV ; Ohashi, J ; Hijikata, M ; Matsushita, I ; Kawana, A ; Yanai, H ; Kirikae, T ; Kuratsuji, T ; Sasazuki, T (Elsevier BV, 2004-10-22)
    We have hypothesized that genetic predisposition influences the progression of SARS. Angiotensin converting enzyme (ACE1) insertion/deletion (I/D) polymorphism was previously reported to show association with the adult respiratory distress syndrome, which is also thought to play a key role in damaging the lung tissues in SARS cases. This time, the polymorphism was genotyped in 44 Vietnamese SARS cases, with 103 healthy controls who had had a contact with the SARS patients and 50 controls without any contact history. SARS cases were divided into either non-hypoxemic or hypoxemic groups. Despite the small sample size, the frequency of the D allele was significantly higher in the hypoxemic group than in the non-hypoxemic group (p=0.013), whereas there was no significant difference between the SARS cases and controls, irrespective of a contact history. ACE1 might be one of the candidate genes that influence the progression of pneumonia in SARS.
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    Polymorphisms of human leucocyte antigen genes in Maonan people in China.
    Ogata, S ; Shi, L ; Matsushita, M ; Yu, L ; Huang, XQ ; Shi, L ; Sun, H ; Ohashi, J ; Muramatsu, M ; Tokunaga, K ; Chu, JY (Wiley, 2007-02)
    We examined human leucocyte antigen (HLA) gene polymorphisms in the Maonan people from southern China. HLA-A, -B and -DRB1 alleles were determined in 108 healthy unrelated Maonan individuals by the polymerase chain reaction-Luminex method, and haplotype frequencies for HLA-A, -B and -DRB1 loci were estimated. The most frequent HLA-A alleles were A*1101 (35.2%), A*0203 (17.6%), A*0207 (13.4%) and A*2402 (13.4%); HLA-B alleles were B*1301(19.9%), B*1502 (14.8%), B*4601 (13.4%) and B*4001 (13.4%); HLA-DRB1 alleles were DRB1*1202 (17.1%), DRB1*1602 (13.0%) and DRB1*1401 (10.7%). The most common haplotypes were A*0207-B*4601 (10.6%), A*1101-B*1301 (10.0%), A*1101-B*4001 (8.4%), B*1502-DRB1*1202 (12.0%), B*4601-DRB1*1401 (5.8%), A*1101-B*1502-DRB1*1202 (7.1%) and A*0207-B*4601-DRB1*1401 (5.3%), profiles that are also found in populations from the southern region of East Asia. Phylogenetic and principal component analyses revealed that the Maonan people belong to the southeastern Asian group and are most closely related to the Buyi people.
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    MxA transcripts with distinct first exons and modulation of gene expression levels by single-nucleotide polymorphisms in human bronchial epithelial cells.
    Noguchi, S ; Hijikata, M ; Hamano, E ; Matsushita, I ; Ito, H ; Ohashi, J ; Nagase, T ; Keicho, N (Springer Science and Business Media LLC, 2013-02)
    Myxovirus resistance A (MxA) is a major interferon (IFN)-inducible antiviral protein. Promoter single-nucleotide polymorphisms (SNPs) of MxA near the IFN-stimulated response element (ISRE) have been frequently associated with various viral diseases, including emerging respiratory infections. We investigated the expression profile of MxA transcripts with distinct first exons in human bronchial epithelial cells. For primary culture, the bronchial epithelium was isolated from lung tissues with different genotypes, and total RNA was subjected to real-time reverse transcription polymerase chain reaction. The previously reported MxA transcript (T1) and a recently registered transcript with a distinct 5' first exon (T0) were identified. IFN-β and polyinosinic-polycytidylic acid induced approximately 100-fold higher expression of the T1 transcript than that of the T0 transcript, which also had a potential ISRE motif near its transcription start site. Even without inducers, the T1 transcript accounted for approximately two thirds of the total expression of MxA, levels of which were significantly associated with its promoter and exon 1 SNPs (rs17000900, rs2071430, and rs464138). Our results suggest that MxA observed in respiratory viral infections is possibly dominated by the T1 transcript and partly influenced by relevant 5' SNPs.
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    Elucidating the origin of HLA-B*73 allelic lineage: Did modern humans benefit by archaic introgression?
    Yasukochi, Y ; Ohashi, J (Springer Science and Business Media LLC, 2017-01)
    A previous study reported that some of the human leukocyte antigen (HLA) alleles and haplotypes in present-day humans were acquired by admixture with archaic humans; specifically, an exceptionally diverged HLA-B*73 allele was proposed to be transmitted from Denisovans, although the DNA sequence of HLA-B*73 has not been detected in the Denisovan genome. Here, we argue against the hypothesis that HLA-B*73 introgressed from Denisovans into early modern humans. A phylogenetic analysis revealed that HLA-B*73:01 formed a monophyletic group with a chimpanzee MHC-B allele, strongly suggesting that the HLA-B*73 allelic lineage has been maintained in humans as well as in chimpanzees since the divergence of humans and chimpanzees. The global distribution of HLA-B*73 allele showed that the population frequency of HLA-B*73 in west Asia (0.24 %)-a possible site of admixture with Denisovans-is lower than that in Europe (0.72 %) and in south Asia (0.69 %). Furthermore, HLA-B*73 is not observed in Melanesia even though the Melanesian genome contains the highest proportion of Denisovan ancestry in present-day human populations. Single nucleotide polymorphisms in HLA-A*11-HLA-C*12:02 or HLA-A*11-C*15 haplotypes, one of which was assumed to be transmitted together with HLA-B*73 from Denisovans by the study of Abi-Rached and colleagues, were not differentiated from those in other HLA-A-C haplotypes in modern humans. These results do not support the introgression hypothesis. Thus, we conclude that it is highly likely that HLA-B*73 allelic lineage has been maintained in the direct ancestors of modern humans.
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    Genetic evidence for contribution of human dispersal to the genetic diversity of EBA-175 in Plasmodium falciparum.
    Yasukochi, Y ; Naka, I ; Patarapotikul, J ; Hananantachai, H ; Ohashi, J (Springer Science and Business Media LLC, 2015-08-01)
    BACKGROUND: The 175-kDa erythrocyte binding antigen (EBA-175) of Plasmodium falciparum plays a crucial role in merozoite invasion into human erythrocytes. EBA-175 is believed to have been under diversifying selection; however, there have been no studies investigating the effect of dispersal of humans out of Africa on the genetic variation of EBA-175 in P. falciparum. METHODS: The PCR-direct sequencing was performed for a part of the eba-175 gene (regions II and III) using DNA samples obtained from Thai patients infected with P. falciparum. The divergence times for the P. falciparum eba-175 alleles were estimated assuming that P. falciparum/Plasmodium reichenowi divergence occurred 6 million years ago (MYA). To examine the possibility of diversifying selection, nonsynonymous and synonymous substitution rates for Plasmodium species were also estimated. RESULTS: A total of 32 eba-175 alleles were identified from 131 Thai P. falciparum isolates. Their estimated divergence time was 0.13-0.14 MYA, before the exodus of humans from Africa. A phylogenetic tree for a large sequence dataset of P. falciparum eba-175 alleles from across the world showed the presence of a basal Asian-specific cluster for all P. falciparum sequences. A markedly more nonsynonymous substitutions than synonymous substitutions in region II in P. falciparum was also detected, but not within Plasmodium species parasitizing African apes, suggesting that diversifying selection has acted specifically on P. falciparum eba-175. CONCLUSIONS: Plasmodium falciparum eba-175 genetic diversity appeared to increase following the exodus of Asian ancestors from Africa. Diversifying selection may have played an important role in the diversification of eba-175 allelic lineages. The present results suggest that the dispersals of humans out of Africa influenced significantly the molecular evolution of P. falciparum EBA-175.
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    Generation of rodent malaria parasites with a high mutation rate by destructing proofreading activity of DNA polymerase δ.
    Honma, H ; Hirai, M ; Nakamura, S ; Hakimi, H ; Kawazu, S-I ; Palacpac, NMQ ; Hisaeda, H ; Matsuoka, H ; Kawai, S ; Endo, H ; Yasunaga, T ; Ohashi, J ; Mita, T ; Horii, T ; Furusawa, M ; Tanabe, K (Oxford University Press (OUP), 2014-08)
    Plasmodium falciparum malaria imposes a serious public health concern throughout the tropics. Although genetic tools are principally important to fully investigate malaria parasites, currently available forward and reverse tools are fairly limited. It is expected that parasites with a high mutation rate can readily acquire novel phenotypes/traits; however, they remain an untapped tool for malaria biology. Here, we generated a mutator malaria parasite (hereinafter called a 'malaria mutator'), using site-directed mutagenesis and gene transfection techniques. A mutator Plasmodium berghei line with a defective proofreading 3' → 5' exonuclease activity in DNA polymerase δ (referred to as PbMut) and a control P. berghei line with wild-type DNA polymerase δ (referred to as PbCtl) were maintained by weekly passage in ddY mice for 122 weeks. High-throughput genome sequencing analysis revealed that two PbMut lines had 175-178 mutations and a 86- to 90-fold higher mutation rate than that of a PbCtl line. PbMut, PbCtl, and their parent strain, PbWT, showed similar course of infection. Interestingly, PbMut lost the ability to form gametocytes during serial passages. We believe that the malaria mutator system could provide a novel and useful tool to investigate malaria biology.
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    A replication study confirms the association of GWAS-identified SNPs at MICB and PLCE1 in Thai patients with dengue shock syndrome.
    Dang, TN ; Naka, I ; Sa-Ngasang, A ; Anantapreecha, S ; Chanama, S ; Wichukchinda, N ; Sawanpanyalert, P ; Patarapotikul, J ; Tsuchiya, N ; Ohashi, J (Springer Science and Business Media LLC, 2014-05-17)
    BACKGROUND: Dengue shock syndrome (DSS), a severe life-threatening form of dengue infection, mostly occurs in children. A recent genome wide association study (GWAS) identified two SNPs, rs3132468 of major histocompatibility complex class I polypeptide-related sequence B (MICB) and rs3765524 of phospholipase C, epsilon 1 (PLCE1), associated with DSS in Vietnamese children. In this study, to examine whether an identical association is found in a different population, the association of these two SNPs with DSS was assessed in Thai children with dengue. METHODS: The rs3132468 and rs3765524 SNPs were genotyped in 917 Thai children with dengue: 76 patients with DSS and 841 patients with non-DSS. The allele frequencies were compared between DSS and non-DSS groups by one-sided Fisher's exact test. The association of rs3132468 and rs3765524 with the mRNA expression levels of MICB and PLCE1 were assessed in EBV-transformed lymphoblastoid cell lines. RESULTS: The reported DSS-risk alleles were significantly associated with DSS in Thai patients with dengue (one-sided P = 0.0213 and odds ratio [OR] = 1.58 for rs3132468-C and one-sided P = 0.0252 and OR = 1.49 for rs3765524-C). The rs3132468-C allele showed a significant association with lower mRNA level of MICB (P = 0.0267), whereas the rs3765524-C allele did not. These results imply that the MICB molecule may play an important role in the prevention of DSS in dengue infection. CONCLUSIONS: Together with previous association studies, we conclude that rs3132468-C at MICB and rs3765524-C at PLCE1 confer risk of DSS in Southeast Asians.
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    Association of the endothelial protein C receptor (PROCR) rs867186-G allele with protection from severe malaria.
    Naka, I ; Patarapotikul, J ; Hananantachai, H ; Imai, H ; Ohashi, J (Springer Science and Business Media LLC, 2014-03-17)
    BACKGROUND: Cytoadhesion of Plasmodium falciparum-infected erythrocytes to endothelial cells in microvessels is a remarkable characteristic of severe malaria. The endothelial protein C receptor (EPCR), encoded by the endothelial protein C receptor gene (PROCR), has recently been identified as an endothelial receptor for specific P. falciparum erythrocyte membrane protein 1 (PfEMP1) subtypes containing domain cassettes (DCs) 8 and 13. The PROCR rs867186-G allele (serine-to-glycine substitution at position 219 of EPCR; 219Gly) has been shown to be associated with higher levels of plasma soluble EPCR (sEPCR). In this study, the association of PROCR rs867186 with severe malaria is examined in Thai population. METHODS: A total of 707 Thai patients with P. falciparum malaria (341 with severe malaria and 336 with mild malaria) were genotyped for rs867186. To assess the association of PROCR rs867186 with severe malaria, three models (dominant, recessive and allelic) were evaluated. The rates of non-synonymous and synonymous substitutions were estimated for the coding sequence of the PROCR gene. RESULTS: The rs867186-GG genotype was significantly associated with protection from severe malaria (P-value=0.026; odds ratio=0.33; 95% confidence interval=0.12-0.90). Evolutionary analysis provided no evidence of strong positive selection acting on the PROCR gene. CONCLUSION: The rs867186-GG genotype showed significant association with protection from severe malaria. The present results suggest that PfEMP1-EPCR interaction, which can mediate cytoadhesion and/or reduce cytoprotective and anti-inflammatory effects, is crucial to the pathogenesis of severe malaria.
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    Diversifying selection on the thrombospondin-related adhesive protein (TRAP) gene of Plasmodium falciparum in Thailand.
    Ohashi, J ; Suzuki, Y ; Naka, I ; Hananantachai, H ; Patarapotikul, J ; Blader, IJ (Public Library of Science (PLoS), 2014)
    Sporozoites of Plasmodium falciparum are transmitted to human hosts by Anopheles mosquitoes. Thrombospondin-related adhesive protein (TRAP) is expressed in sporozoites and plays a crucial role in sporozoite gliding and invasion of human hepatocytes. A previous study showed that the TRAP gene has been subjected to balancing selection in the Gambian P. falciparum population. To further study the molecular evolution of the TRAP gene in Plasmodium falciparum, we investigated TRAP polymorphisms in P. falciparum isolates from Suan Phueng District in Ratchaburi Province, Thailand. The analysis of the entire TRAP coding sequences in 32 isolates identified a total of 39 single nucleotide polymorphisms (SNPs), which comprised 37 nonsynonymous and two synonymous SNPs. McDonald-Kreitman test showed that the ratio of the number of nonsynonymous to synonymous polymorphic sites within P. falciparum was significantly higher than that of the number of nonsynonymous to synonymous fixed sites between P. falciparum and P. reichenowi. Furthermore, the rate of nonsynonymous substitution was significantly higher than that of synonymous substitution within Thai P. falciparum. These results indicate that the TRAP gene has been subject to diversifying selection in the Thai P. falciparum population as well as the Gambian P. falciparum population. Comparison of our P. falciparum isolates with those from another region of Thailand (Tak province, Thailand) revealed that TRAP was highly differentiated between geographically close regions. This rapid diversification seems to reflect strong recent positive selection on TRAP. Our results suggest that the TRAP molecule is a major target of the human immune response to pre-erythrocytic stages of P. falciparum.