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        Population-Based Molecular Epidemiology of Leprosy in Cebu, Philippines

        Sakamuri, R. M.,Kimura, M.,Li, W.,Kim, H.-C.,Lee, H.,Kiran, M. D.,Black, W. C.,Balagon, M.,Gelber, R.,Cho, S.-N.,Brennan, P. J.,Vissa, V. American Society for Microbiology 2009 Journal of clinical microbiology Vol.47 No.9

        <P>To address the persisting problem of leprosy in Cebu, Philippines, we compiled a database of more than 200 patients who attend an established referral skin clinic. We described the patient characteristics in conventional demographic parameters and also applied multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA) and single nucleotide polymorphism (SNP) typing for Mycobacterium leprae in biopsied skin lesion samples. These combined approaches revealed that transmission is ongoing, with the affected including the young Cebuano population under 40 years of age in both crowded cities and rural areas of the island. The emergence of multicase families (MCF) is indicative of infection unconstrained by standard care measures. For the SNPs, we designed a low-cost PCR-restriction fragment length polymorphism typing method. MLVA in M. leprae was highly discriminatory in this population yet could retain broad groups, as defined by the more stable SNPs, implying temporal marker stability suitable for interpreting population structures and evolution. The majority of isolates belong to an Asian lineage (SNP type 1), and the rest belong to a putative postcolonial lineage (SNP type 3). Specific alleles at two VNTR loci, (GGT)5 and 21-3, were highly associated with SNP type 3 in this population. MLVA identified M. leprae genotype associations for patients with known epidemiological links such as in MCFs and in some villages. These methods provide a molecular database and a rational framework for targeted approaches to search and confirm leprosy transmission in various scenarios.</P>

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        Density, ultrasonic velocity, viscosity and their excess parameters of the binary mixtures of N,N-dimethylaniline+1-alkanols (C3-C5), +2-alkanols (C3-C4), +2-methyl-1-propanol, +2-methyl-2-propanol at 303.15K

        Sivarambabu Sakamuri,Gowrisankar Manukonda,Venkateswarlu Ponneri,Sivakumar Kasibhatta 한국화학공학회 2013 Korean Journal of Chemical Engineering Vol.30 No.5

        The density, ultrasonic velocity of sound and viscosity of binary mixtures of N,N-dimethyl aniline (N,NDMA)with 1-propanol, +2-propanol, +1-butanol, +2-butanol, +1-pentanol, +2-methyl-1-propanol, +2-methyl-2-propanol were measured at 303.15 K. These experimental data have been used to calculate excess volume V E, excess ultrasonic speeds uE, excess intermolecular free length Lf E, excess acoustic impedance Z E, excess isentropic compressibility κs E, deviation in viscosity Δη and excess Gibbs free energy of activation of viscous flow (G*E ). The values of Lf Eand κs E are negative over the wide range of composition for all the binary mixtures, while the values of Z Eare positive. These results have been used to discuss the nature of interaction between unlike molecules in terms of hydrogen bonding, dipole-dipole interaction, proton-acceptor interaction and dispersive forces. The viscosity data have been correlated using three equations: Grunberg and Nissan, Katti & Chaudhri and Hind et al. The excess/deviations were fitted by a Redlich-Kister equation and the results were analyzed in terms of specific interactions present in these mixtures.

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        Rapid Variable-Number Tandem-Repeat Genotyping for Mycobacterium leprae Clinical Specimens

        Kimura, M.,Sakamuri, R. M.,Groathouse, N. A.,Rivoire, B. L.,Gingrich, D.,Krueger-Koplin, S.,Cho, S.-N.,Brennan, P. J.,Vissa, V. American Society for Microbiology 2009 Journal of clinical microbiology Vol.47 No.6

        <P>Mycobacterium leprae is the noncultivable pathogen of leprosy. Since the genome sequence of an isolate of M. leprae has become available, multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA) has been explored as a tool for strain typing and identification of chains of transmission of leprosy. In order to discover VNTRs and develop methods transferable to clinical samples, MLVA was applied to a global collection of M. leprae isolates derived from leprosy patients and propagated in armadillo hosts. PCR amplification, agarose gel electrophoresis, and sequencing methods were applied to DNA extracts from these infected armadillo tissues (n = 21). We identified polymorphisms in 15 out of 25 short-tandem-repeat (STR) loci previously selected by in silico analyses of the M. leprae genome. We then developed multiplex PCR for amplification of these 15 loci in four separate PCRs suitable for fluorescent fragment length analysis and demonstrated STR profiles highly concordant with those from the sequencing methods. Subsequently, we extended this method to DNA extracts from human clinical specimens, such as skin biopsy specimens (n = 30). With these techniques, mapping of multiple loci and differentiation of genotypes have been possible using total DNA extracts from limited amounts of clinical samples at a reduced cost and with less time. These practical methods are therefore available and applicable to answer focused epidemiological questions and to allow monitoring of the transmission of M. leprae in different countries where leprosy is endemic.</P>

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