bluuebunny/crossref_metadata_2025_split huggingface dataset:
PCR-Stop analysis as a new tool for qPCR assay validation
Authors: Witte Anna Kristina , Mester Patrick , Fister Susanne , Süß Beate , Wagner Martin , Rossmanith Peter 
Published: May 2018

AbstractProgressively more qPCR assays have been developed in recent years in numerous fields of application. These assays are routinely validated using calibration curves, but essential validation per se such as Poisson analysis is frequently neglected. However, validation is crucial for determination of resolution and quantitative and qualitative limits. The new test method PCR-Stop analysis presented in this work investigates assay performance during initial qPCR cycles. PCRs with one to five pre-runs are performed while the subsequent main qPCR runs reflect pre-run replication rates. Ideally, DNA doubles according to pre-runs, there is no variation between replicates and qPCR starts immediately at the first cycle with its average efficiency. This study shows two exemplary qPCR assays, both with suitable calibration curves and efficiencies. We demonstrated thereby the benefits of PCR-Stop analysis revealing quantitative and qualitative resolution of both assays, the limits of one of those assays and thus avoiding misinterpretations in qPCR analysis. Furthermore, data displayed that a well performing assay starts indeed with its average efficiency.
Nanopore sequencing of drug-resistance-associated genes in malaria parasites, Plasmodium falciparum
Authors: Runtuwene Lucky R. , Tuda Josef S. B. , Mongan Arthur E. , Makalowski Wojciech , Frith Martin C. , Imwong Mallika , Srisutham Suttipat , Nguyen Thi Lan Anh , Tuan Nghia Nguyen , Eshita Yuki , Maeda Ryuichiro , Yamagishi Junya , Suzuki Yutaka 
Published: May 2018

AbstractHere, we report the application of a portable sequencer, MinION, for genotyping the malaria parasite Plasmodium falciparum. In the present study, an amplicon mixture of nine representative genes causing resistance to anti-malaria drugs is diagnosed. First, we developed the procedure for four laboratory strains (3D7, Dd2, 7G8, and K1), and then applied the developed procedure to ten clinical samples. We sequenced and re-sequenced the samples using the obsolete flow cell R7.3 and the most recent flow cell R9.4. Although the average base-call accuracy of the MinION sequencer was 74.3%, performing >50 reads at a given position improves the accuracy of the SNP call, yielding a precision and recall rate of 0.92 and 0.8, respectively, with flow cell R7.3. These numbers increased significantly with flow cell R9.4, in which the precision and recall are 1 and 0.97, respectively. Based on the SNP information, the drug resistance status in ten clinical samples was inferred. We also analyzed K13 gene mutations from 54 additional clinical samples as a proof of concept. We found that a novel amino-acid changing variation is dominant in this area. In addition, we performed a small population-based analysis using 3 and 5 cases (K13) and 10 and 5 cases (PfCRT) from Thailand and Vietnam, respectively. We identified distinct genotypes from the respective regions. This approach will change the standard methodology for the sequencing diagnosis of malaria parasites, especially in developing countries.
A phase I single-blind clinical trial to evaluate the safety of oil palm phenolics (OPP) supplementation in healthy volunteers
Authors: Fairus Syed , Leow Soon-Sen , Mohamed Isa Naina , Tan Yew-Ai , Sundram Kalyana , Sambanthamurthi Ravigadevi 
Published: May 2018

AbstractPlant phenolics are being increasingly consumed globally with limited scientific and clinical evidence pertaining to safety and efficacy. The oil palm fruit contains a cocktail of phenolics, and palm oil production results in high volumes of aqueous by-products enriched in phenolics and bioactives. Several lines of evidence from in vitro and in vivo animal studies confirmed that the aqueous extract enriched in phenolics and other bioactives collectively known as oil palm phenolics (OPP) is safe and has potent bioactivity. A phase one clinical trial was conducted to evaluate the safety and effects of OPP in healthy volunteers. In this single-blind trial, 25 healthy human volunteers were supplemented with 450 mg gallic acid equivalent (GAE)/day of OPP or control treatments for a 60-day period. Fasting blood and urine samples were collected at days 1, 30 and 60. Medical examination was performed during these trial interventions. All clinical biochemistry profiles observed throughout the control and OPP treatment period were in the normal range with no major adverse effect (AE) or serious adverse effect (SAE) observed. Additionally, OPP supplementation resulted in improvement of total cholesterol and LDL-C levels, compared to the control treatment. The outcomes support our previous observations that OPP is safe and may have a protective role in reducing cholesterol levels.
Molecular Evidence of Genome Editing in a Mouse Model of Immunodeficiency
Authors: Abdul-Razak H. H. , Rocca C. J. , Howe S. J. , Alonso-Ferrero M. E. , Wang J. , Gabriel R. , Bartholomae C. C. , Gan C. H. V. , Garín M. I. , Roberts A. , Blundell M. P. , Prakash V. , Molina-Estevez F. J. , Pantoglou J. , Guenechea G. , Holmes M. C. , Gregory P. D. , Kinnon C. , von Kalle C. , Schmidt M. , Bueren J. A. , Thrasher A. J. , Yáñez-Muñoz R. J. 
Published: May 2018

AbstractGenome editing is the introduction of directed modifications in the genome, a process boosted to therapeutic levels by designer nucleases. Building on the experience of ex vivo gene therapy for severe combined immunodeficiencies, it is likely that genome editing of haematopoietic stem/progenitor cells (HSPC) for correction of inherited blood diseases will be an early clinical application. We show molecular evidence of gene correction in a mouse model of primary immunodeficiency. In vitro experiments in DNA-dependent protein kinase catalytic subunit severe combined immunodeficiency (Prkdc scid) fibroblasts using designed zinc finger nucleases (ZFN) and a repair template demonstrated molecular and functional correction of the defect. Following transplantation of ex vivo gene-edited Prkdc scid HSPC, some of the recipient animals carried the expected genomic signature of ZFN-driven gene correction. In some primary and secondary transplant recipients we detected double-positive CD4/CD8 T-cells in thymus and single-positive T-cells in blood, but no other evidence of immune reconstitution. However, the leakiness of this model is a confounding factor for the interpretation of the possible T-cell reconstitution. Our results provide support for the feasibility of rescuing inherited blood disease by ex vivo genome editing followed by transplantation, and highlight some of the challenges.
Homoharringtonine induced immune alteration for an Efficient Anti-tumor Response in Mouse Models of Non-small Cell Lung Adenocarcinoma Expressing Kras Mutation
Authors: Weng Tzu-Yang , Wu Hsuan Franziska , Li Chung-Yen , Hung Yu-Hsuan , Chang Yu-Wei , Chen Yi-Ling , Hsu Hui-Ping , Chen Yu-Hung , Wang Chih-Yang , Chang Jang-Yang , Lai Ming-Derg 
Published: May 2018

AbstractHomoharringtonine (HHT), an inhibitor of protein synthesis, has been used to treat leukemia. Its therapeutic effects on non-small cell lung adenocarcinoma carrying KRAS mutation and their immune system are less understood. The present study examined the therapeutic efficacy and the immune effects of HHT in two murine lung tumor models, xenograft and transgenic, carrying the Kras mutation G12D and G12C respectively. HHT exhibited efficient anticancer activity, significantly suppressing lung tumor growthin vitroandin vivo. The levels of 22 cytokines and chemokines in splenocytes of tumor-bearing mice were examined. Interleukin-12 expression was lower in splenocytes of HHT-treated mice when compared to the controls as demonstrated by a cytokine array and an enzyme-linked immunosorbent assay. The expression levels of CD80, CD86, and CD69 in B220+B cells from splenocytes of HHT-treated mice were higher than that of control mice in two mouse tumor models. Furthermore, antitumor effect of HHT was attenuated with depletion of B cells. Increased numbers of CD80+and CD86+B cells were observed in the mice treated with narciclasine, another translation inhibitor. In conclusion, HHT changed the features of immune cells, and exhibited efficient anti-tumor activity against lung tumor carrying mutant Kras expression.
A comprehensive analysis of e-CAS cell line reveals they are mouse macrophages
Authors: Evans Elizabeth , Paillot Romain , López-Álvarez María Rocío 
Published: May 2018

AbstractThe 3Rs principles (Replacement, Reduction and Refinement) are focused on finding alternatives to the use of animals in research. In this regard, cell lines are popular and useful tools for the replacement of primary cells in in vitro studies. However, around 15–30% of cell lines used in research have been misidentified or cross-contaminated generating concerns about the results obtained from experiments that use them. Here we described how old aliquots of an equine macrophage cell line (e-CAS) stored at the Animal Health Trust did not contain equine cells but macrophages of murine origin (m-CAS).
A new genetic mechanism of natural gas accumulation
Authors: Yang Chengyu , Ni Zhiyong , Wang Tieguan , Chen Zhonghong , Hong Haitao , Wen Long , Luo Bing , Wang Wenzhi 
Published: May 2018

AbstractNatural gas of organic origin is primarily biogenic or thermogenic; however, the formation of natural gas is occasionally attributed to hydrothermal activity. The Precambrian dolomite reservoir of the Anyue gas field is divided into three stages. Dolomite-quartz veins were precipitated after two earlier stages of dolomite deposition. Fluid inclusions in the dolomite and quartz are divided into pure methane (P-type), methane-bearing (M-type), aqueous (W-type), and solid bitumen-bearing (S-type) inclusions. The W-type inclusions within the quartz and buried dolomite homogenized between 107 °C and 223 °C. Furthermore, the trapping temperatures and pressures of the fluid (249 °C to 319 °C and 1619 bar to 2300 bar, respectively) are obtained from the intersections of the isochores of the P-type and the coeval W-type inclusions in the quartz. However, the burial history of the reservoir indicates that the maximum burial temperature did not exceed 230 °C. Thus, the generation of the natural gas was not caused solely by the burial of the dolomite reservoir. The results are also supported by the presence of paragenetic pyrobitumen and MVT lead-zinc ore. A coupled system of occasional invasion by hydrothermal fluids and burial of the reservoir may represent a new genetic model for natural gas accumulation in this gas field.
Influence of Household Water Filters on Bacteria Growth and Trace Metals in Tap Water of Doha, Qatar
Authors: Nriagu Jerome , Xi Chuanwu , Siddique Azhar , Vincent Annette , Shomar Basem 
Published: May 2018

AbstractDeteriorating water quality from aging infrastructure, growing threat of pollution from industrialization and urbanization, and increasing awareness about waterborne diseases are among the factors driving the surge in worldwide use of point-of-entry (POE) and point-of-use (POU) filters. Any adverse influence of such consumer point-of-use systems on quality of water at the tap remains poorly understood, however. We determined the chemical and microbiological changes in municipal water from the point of entry into the household plumbing system until it leaves from the tap in houses equipped with filters. We show that POE/POU devices can induce significant deterioration of the quality of tap water by functioning as traps and reservoirs for sludge, scale, rust, algae or slime deposits which promote microbial growth and biofilm formation in the household water distribution system. With changes in water pressure and physical or chemical disturbance of the plumbing system, the microorganisms and contaminants may be flushed into the tap water. Such changes in quality of household water carry a potential health risk which calls for some introspection in widespread deployment of POE/POU filters in water distribution systems.
The bioeroding sponge Cliona orientalis will not tolerate future projected ocean warming
Authors: Ramsby Blake D. , Hoogenboom Mia O. , Smith Hillary A. , Whalan Steve , Webster Nicole S. 
Published: May 2018

AbstractCoral reefs face many stressors associated with global climate change, including increasing sea surface temperature and ocean acidification. Excavating sponges, such as Cliona spp., are expected to break down reef substrata more quickly as seawater becomes more acidic. However, increased bioerosion requires that Cliona spp. maintain physiological performance and health under continuing ocean warming. In this study, we exposed C. orientalis to temperature increments increasing from 23 to 32 °C. At 32 °C, or 3 °C above the maximum monthly mean (MMM) temperature, sponges bleached and the photosynthetic capacity of Symbiodinium was compromised, consistent with sympatric corals. Cliona orientalis demonstrated little capacity to recover from thermal stress, remaining bleached with reduced Symbiodinium density and energy reserves after one month at reduced temperature. In comparison, C. orientalis was not observed to bleach during the 2017 coral bleaching event on the Great Barrier Reef, when temperatures did not reach the 32 °C threshold. While C. orientalis can withstand current temperature extremes (<3 °C above MMM) under laboratory and natural conditions, this species would not survive ocean temperatures projected for 2100 without acclimatisation or adaptation (≥3 °C above MMM). Hence, as ocean temperatures increase above local thermal thresholds, C. orientalis will have a negligible impact on reef erosion.
Investigation of dose-rate effects and cell-cycle distribution under protracted exposure to ionizing radiation for various dose-rates
Authors: Matsuya Yusuke , McMahon Stephen J. , Tsutsumi Kaori , Sasaki Kohei , Okuyama Go , Yoshii Yuji , Mori Ryosuke , Oikawa Joma , Prise Kevin M. , Date Hiroyuki 
Published: May 2018

AbstractDuring exposure to ionizing radiation, sub-lethal damage repair (SLDR) competes with DNA damage induction in cultured cells. By virtue of SLDR, cell survival increases with decrease of dose-rate, so-called dose-rate effects (DREs). Here, we focused on a wide dose-rate range and investigated the change of cell-cycle distribution during X-ray protracted exposure and dose-response curves via hybrid analysis with a combination of in vitro experiments and mathematical modelling. In the course of flow-cytometric cell-cycle analysis and clonogenic assays, we found the following responses in CHO-K1 cells: (1) The fraction of cells in S phase gradually increases during 6 h exposure at 3.0 Gy/h, which leads to radio-resistance. (2) Slight cell accumulation in S and G2/M phases is observed after exposure at 6.0 Gy/h for more than 10 hours. This suggests that an increase of SLDR rate for cells in S phase during irradiation may be a reproducible factor to describe changes in the dose-response curve at dose-rates of 3.0 and 6.0 Gy/h. By re-evaluating cell survival for various dose-rates of 0.186–60.0 Gy/h considering experimental-based DNA content and SLDR, it is suggested that the change of S phase fraction during irradiation modulates the dose-response curve and is possibly responsible for some inverse DREs.
Association of a rare variant of the TNFSF13B gene with susceptibility to Rheumatoid Arthritis and Systemic Lupus Erythematosus
Authors: González-Serna David , Ortiz-Fernández Lourdes , Vargas Sofía , García Antonio , Raya Enrique , Fernández-Gutierrez Benjamín , López-Longo Francisco Javier , Balsa Alejandro , González-Álvaro Isidoro , Narvaez Javier , Gómez-Vaquero Carmen , Sabio José Mario , García-Portales Rosa , González-Escribano María Francisca , Tolosa Carles , Carreira Patricia , Kiemeney Lambertus , Coenen Marieke J. H. , Witte Torsten , Schneider Matthias , González-Gay Miguel Ángel , Martín Javier 
Published: May 2018

AbstractA rare variant (BAFF-var) of the tumor necrosis factor superfamily 13b (TNFSF13B) gene has been recently associated with multiple sclerosis (MS) and systemic lupus erythematosus (SLE). The aim of this study was to investigate the association between TNFSF13B BAFF-var and susceptibility to rheumatoid arthritis (RA) and replicate that association in SLE. 6,218 RA patients, 2,575 SLE patients and 4,403 healthy controls from three different countries were included in the study. TNFSF13B BAFF-var was genotyped using TaqMan allelic discrimination assay. PLINK software was used for statistical analyses. TNFSF13B BAFF-var was significantly associated with RA (p = 0.015, OR = 1.21, 95% CI = 1.03–1.41) in the Spanish cohort. A trend of association was observed in the Dutch (p = 0.115) and German (p = 0.228) RA cohorts. A meta-analysis of the three RA cohorts included in this study revealed a statistically significant association (p = 0.002, OR = 1.24, 95% CI = 1.10–1.38). In addition, TNFSF13B BAFF-var was significantly associated with SLE in the Spanish (p = 0.001, OR = 1.41, 95% CI = 1.14–1.74) and the German cohorts (p = 0.030, OR = 1.86, 95% CI = 1.05–3.28), with a statistically significant p-value obtained in the meta-analysis (p = 0.0002, OR = 1.46, 95% CI = 1.09–2.32). The results obtained confirm the known association of TNFSF13B BAFF-var with SLE and, for the first time, demonstrate that this variant contributes to susceptibility to RA.
Sub-threshold signal encoding in coupled FitzHugh-Nagumo neurons
Authors: Masoliver Maria , Masoller Cristina 
Published: May 2018

AbstractDespite intensive research, the mechanisms underlying the neural code remain poorly understood. Recent work has focused on the response of a single neuron to a weak, sub-threshold periodic signal. By simulating the stochastic FitzHugh-Nagumo (FHN) model and then using a symbolic method to analyze the firing activity, preferred and infrequent spike patterns (defined by the relative timing of the spikes) were detected, whose probabilities encode information about the signal. As not individual neurons but neuronal populations are responsible for sensory coding and information transfer, a relevant question is how a second neuron, which does not perceive the signal, affects the detection and the encoding of the signal, done by the first neuron. Through simulations of two stochastic FHN neurons we show that the encoding of a sub-threshold signal in symbolic spike patterns is a plausible mechanism. The neuron that perceives the signal fires a spike train that, despite having an almost random temporal structure, has preferred and infrequent patterns which carry information about the signal. Our findings could be relevant for sensory systems composed by two noisy neurons, when only one detects a weak external input.
The evolution of genomic and epigenomic features in two Pleurotus fungi
Authors: Zhang Zhibin , Wen Jiawei , Li Juzuo , Ma Xintong , Yu Yanan , Tan Xiao , Wang Qi , Liu Bao , Li Xiaomeng , Li Yu , Gong Lei 
Published: May 2018

AbstractPleurotus tuoliensis (Bailinggu, designated Pt) and P. eryngii var. eryngii (Xingbaogu, designated Pe) are highly valued edible mushrooms. We report de novo assemblies of high-quality genomes for both mushrooms based on PacBio RS II sequencing and annotation of all identified genes. A comparative genomics analysis between Pt and Pe with P. ostreatus as an outgroup taxon revealed extensive genomic divergence between the two mushroom genomes primarily due to the rapid gain of taxon-specific genes and disruption of synteny in either taxon. The re-appraised phylogenetic relationship between Pt and Pe at the genome-wide level validates earlier proposals to designate Pt as an independent species. Variation of the identified wood-decay-related gene content can largely explain the variable adaptation and host specificity of the two mushrooms. On the basis of the two assembled genome sequences, methylomes and the regulatory roles of DNA methylation in gene expression were characterized and compared. The genome, methylome and transcriptome data of these two important mushrooms will provide valuable information for advancing our understanding of the evolution of Pleurotus and related genera and for facilitating genome- and epigenome-based strategies for mushroom breeding.
Resolving the smell of wood - identification of odour-active compounds in Scots pine (Pinus sylvestris L.)
Authors: Schreiner Linda , Bauer Patrick , Buettner Andrea 
Published: May 2018

AbstractBeing one of the most common trees in forests, Pinus sylvestris L. is a frequently used raw material for wood products. Its specific odour is, however, mostly unresolved to date. Accordingly, we investigated Scots pine wood samples grown in Germany for their main odorant composition. We employed dedicated odorant analysis techniques such as gas chromatography-olfactometry (GC-O) and aroma extract dilution analysis (AEDA) and successfully detected 44 odour-active compounds; of these, 39 substances were successfully identified by gas chromatography-mass spectrometry/olfactometry (GC-MS/O) and two-dimensional gas chromatography-mass spectrometry/olfactometry (2D-GC-MS/O). Among the main odorants found were (E,E)-nona-2,4-dienal, vanillin, phenylacetic acid, 3-phenylpropanoic acid, δ-octalactone and α-pinene, all of them having been detected with high flavour dilution factors during GC-O analyses. The majority of the identified odorants were fatty acid degradation products, plus some terpenoic substances and odorous substances resulting from the degradation of lignin. Although some of the detected substances have previously been reported as constituents of wood, 11 substances are reported here for the first time as odour-active compounds in wood, amongst them heptanoic acid, γ-octalactone, δ-nonalactone and (E,Z,Z)-trideca-2,4,7-trienal.
Exploration of an urban lake management model to simulate chlorine interference based on the ecological relationships among aquatic species
Authors: Yan Zhiqiang , Wang Yafei , Wu Di , Xia Beicheng 
Published: May 2018

AbstractIn eutrophic lakes, algae are known to be sensitive to chlorine, but the impact of chlorine on the wider ecosystem has not been investigated. To quantitatively investigate the effects of chlorine on the urban lake ecosystem and analyze the changes in the aquatic ecosystem structure, a dynamic response model of aquatic species to chlorine was constructed based on the biomass density dynamics of aquatic species of submerged macrophytes, phytoplankton, zooplankton, periphyton, and benthos. The parameters were calibrated using data from the literature and two simulative experiments. The model was then validated using field data from an urban lake with a surface area of approximately 8000 m2 located in the downtown area of Guangzhou, South China. The correlation coefficient (R), root mean square error-observations standard deviation ratio (RSR) and index of agreement (IOA) were used to evaluate the accuracy and reliability of the model and the results were consistent with the observations (0.446 R < 0.985, RSR < 0.7, IOA > 0.6). Comparisons between the simulated and observed trends confirmed the feasibility of using this model to investigate the dynamics of aquatic species under chlorine interference. The model can help managers apply a modest amount of chlorine to control eutrophication and provides scientific support for the management of urban lakes.