Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance

The polymerase chain reaction (PCR) is widely used for applications which require a high level of specificity and reliability, such as genetic testing, clinical diagnostics, blood screening, forensics and biodefense. Great improvements to PCR performance have been achieved by the use of Hot Start activation strategies that aim to prevent DNA polymerase extension until more stringent, higher temperatures are reached. Herein we present a novel Hot Start activation approach in PCR where primers contain one or two thermolabile, 4-oxo-1-pentyl (OXP) phosphotriester (PTE) modification groups at 3′-terminal and 3′-penultimate internucleotide linkages. Studies demonstrated that the presence of one or more OXP PTE modifications impaired DNA polymerase primer extension at the lower temperatures that exist prior to PCR amplification. Furthermore, incubation of the OXP-modified primers at elevated temperatures was found to produce the corresponding unmodified phosphodiester (PDE) primer, which was then a suitable DNA polymerase substrate. The OXP-modified primers were tested in conventional PCR with endpoint detection, in one-step reverse transcription (RT)–PCR and in real-time PCR with SYBR Green I dye and Taqman® probe detection. When OXP-modified primers were used as substitutes for unmodified PDE primers in PCR, significant improvement was observed in the specificity and efficiency of nucleic acid target amplification

La traducción y los textos bilingües (japonés-inglés) para estudiantes de japonés : análisis de las antologías de relatos cortos y de las revistas de idioma

El treball estudia els dos tipus de textos bilingües japonès-anglès (llibres i revistes) que existeixen per a estudiants de japonès. La primera part analitza quatre antologies des de la perspectiva de la paratextualitat i de la teoria funcionalista de Christiane Nord (1997). A continuació, s'estudien les tècniques de traducció, a partir dels desenvolupaments teòrics d'Amparo Hurtado i Lucía Molina (2002), en el conte «The Third Night» de Natsume Sôseki. A la segona part s'investiguen dues revistes d'idiomes mensuals, Nihongo Journal i Hiragana Times. Es tracta d'analitzar la influència del propòsit pedagògic en les traduccions. Se segueix el procediment ja emprat a la primera part, exceptuant l'anàlisi de les tècniquesEn el trabajo se tratan los dos tipos de textos bilingües (libros y revistas) japonés-inglés disponibles para estudiantes de japonés. En la primera parte del trabajo se analizan cuatro antologías teniendo en cuenta la paratextualidad y usando la teoría funcionalista de Christiane Nord (1997). A continuación, se estudian las técnicas de traducción en el cuento «The Third Night» de Natsume Sôseki, a partir de los desarrollos teóricos de Amparo Hurtado y Lucía Molina (2002). En la segunda parte se investiga la influencia del propósito pedagógico en las traducciones de dos revistas de idioma mensuales, Nihongo Journal y Hiragana Times. Se sigue el mismo procedimiento que el empleado en la primera parte, exceptuando el análisis de las técnicasThis thesis treats two types of Japanese-English bilingual texts (anthologies and magazines) available for those who study Japanese. The first part analyzes four anthologies on paratextuality and then by using functionalist approach of Christiane Nord (1997). Finally, the translation techniques applied in the short story "The Third Night" by Natsume Sôseki are studied based on the theoretical developments of Amparo Hurtado and Lucía Molina (2002). The second part examines the influence of pedagogical purpose on two kinds of monthly language magazines: Nihongo Journal and Hiragana Times. The procedure of the first part is employed except for the analysis of translation technique

Dipole-quadrupole interactions and the nature of phase III of compressed hydrogen

A new class of strongly infrared active structures is identified for phase III of compressed molecular H2 by constant-pressure ab initio molecular dynamics and density-functional perturbation calculations. These are planar quadrupolar structures obtained as a distortion of low-pressure quadrupolar phases, after they become unstable at about 150 GPa due to a zone-boundary soft phonon. The nature of the II-III transition and the origin of the IR activity are rationalized by means of simple electrostatics, as the onset of a stabilizing dipole-quadrupole interaction.Comment: 4 pages, 3 figures. To appear in Phys. Rev. Let

Quest for a potent antimalarial drug lead: synthesis and evaluation of 6,7-dimethoxyquinazoline-2,4-diamines

Quinazolines have long been known to exert varied pharmacologic activities that make them suitable for use in treating hypertension, viral infections, tumors, and malaria. Since 2014, we have synthesized approximately 150 different 6,7-dimethoxyquinazoline-2,4-diamines and evaluated their antimalarial activity via structure-activity relationship studies. Here, we summarize the results and report the discovery of 6,7-dimethoxy-N(4)-(1-phenylethyl)-2-(pyrrolidin-1-yl)quinazolin-4-amine (20, SSJ-717), which exhibits high antimalarial activity as a promising antimalarial drug lead

Mosquito Biodiversity Patterns Around Urban Environments in South-Central Okinawa Island, Japan

Okinawa is the largest, most urbanized, and densely populated island in the Ryukyus Archipelago, where mosquito species diversity has been thoroughly studied. However, the south-central Okinawa mosquito fauna has been relatively poorly studied. Here, we present results from a mosquito faunal survey in urban environments of Nishihara city, south-central Okinawa. Mosquitoes were sampled biweekly, from April 2007 to March 2008, at 3 different environments: a forest preserve, an animal farm, and a water reservoir. We employed 4 mosquito collection methods: 1) oviposition traps; 2) light traps; 3) sweep nets; and 4) larval surveys of tree holes, leaf axils, and artificial water containers. We collected a total of 568 adults and 10,270 larvae belonging to 6 genera and 13 species, including 6 species of medical importance: Aedes albopictus, Armigeres subalbatus, Anopheles Hyrcanus group, Culex bitaeniorhynchus, Cx. quinquefasciatus, and Cx. tritaeniorhynchus. Mosquito species composition was similar to data from previous studies in Okinawa Island. The flattening of the species accumulation curve suggests that our diversity sampling was exhaustive with light and oviposition traps, as well as the coincidence between the species richness we found in the field and estimates from the Chao2 index, a theoretical estimator of species richness based on species abundance. This study highlights the importance of combining several sampling techniques to properly characterize regional mosquito fauna and to monitor changes in the presence of mosquito species

Isomeric excitation energy for 99^{99}Inm^{m} from mass spectrometry reveals constant trend next to doubly magic 100^{100}Sn

The excitation energy of the 1/2$^-$ isomer in $^{99}$In at ${N=50}$ is measured to be 671(37) keV and the mass uncertainty of the 9/2$^+$ ground state is significantly reduced using the ISOLTRAP mass spectrometer at ISOLDE/CERN. The measurements exploit a major improvement in the resolution of the multi-reflection time-of-flight mass spectrometer. The results reveal an intriguing constancy of the $1/2^-$ isomer excitation energies in neutron-deficient indium that persists down to the $N = 50$ shell closure, even when all neutrons are removed from the valence shell. This trend is used to test large-scale shell model, \textit{ab initio}, and density functional theory calculations. The models have difficulties describing both the isomer excitation energies and ground-state electromagnetic moments along the indium chain.Comment: 13 pages, 4 figure

Comprehensive biomarker analyses identifies HER2, EGFR, MET RNA expression and thymidylate synthase 5'UTR SNP as predictors of benefit from S-1 adjuvant chemotherapy in Japanese patients with stage II/III gastric cancer

Purpose: A comprehensive molecular analysis was conducted to identify prognostic and predictive markers for adjuvant S-1 chemotherapy in stage II/III Japanese gastric cancer (GC) patients and to evaluate their potential suitability for alternative cytotoxic or targeted drugs. Experimental Design: We investigated genetic polymorphisms of enzymes potentially involved in 5-fluoruracil (5-FU) metabolism as well as platinum resistance, previously identified genomic subtypes potentially predicting 5-FU benefit, and mRNA expression levels of receptor tyrosine kinases and KRAS as potential treatment targets in a single institution cohort of 252 stage II/III GC patients treated with or without S-1 after D2 gastrectomy. Results: 88% and 62% GC had a potentially 5-FU sensitive phenotype by SNP analyses of TS 3'UTR, and TS 5'UTR, respectively. 24%, 46%, 40%, 5%, and 44% GC had a potentially platinum sensitive phenotype by SNP analyses of GSTP1, ERCC1 rs11615, ERCC1 rs3212986, ERCC2, and XRCC1, respectively. High HER2, EGFR, FGFR2, or MET mRNA expression was observed in 49%, 66%, 72%, and 54% GC, respectively. High HER2 expression was the only significant prognosticator (HR=3.912, 95%CI: 1.706-8.973, p=0.0005). High HER2 (p=0.031), low EGFR (p=0.124), high MET (p=0.165) RNA expression, and TS 5'UTR subtype 2R/2R, 2R/3C, or 3C (p=0.058) were significant independent predictors for S-1 resistance. Conclusions: The present study suggests that platinum-based or RTK targeted agents could be alternative treatment options for a substantial subgroup of Japanese GC patients currently treated with S-1. HER2, EGFR, MET, and TS 5'UTR SNP appear to be promising predictive markers for S-1 resistance warranting validation in an independent GC series