bluuebunny/crossref_metadata_2025 huggingface dataset:
Real-Time Estimation and Calibration of GLONASS Inter-Frequency Phase and Code Bias
Authors: Yang Zhixin , Liu Hui , Lou Yidong , Shu Bao , Xu Longwei , Wang Yifei , Xie Baofei
Published: December 2019

The frequency division multiple access (FDMA) strategy used in GLONASS causes inter-frequency phase bias (IFPB) and inter-frequency code bias (IFCB) between receivers from different manufacturers. The existence of IFPB and IFCB significantly increases the difficulties of fixing GLONASS ambiguity and limits the accuracy and reliability of GLONASS positioning. Moreover, the initial value of IFPB and IFCB may be unavailable or unreliable with the increasing number of receivers from different manufacturers in recent years. In this study, a real-time and reliable calibration algorithm of IFPB and IFCB based on multi-GNSS assistance is proposed by providing a fixed solution. Real-time IFPB rate and IFCB can be obtained using this algorithm without the initial IFPB and IFCB. The IFPB rate for all GLONASS satellites and IFCB for each GLONASS satellite are estimated due to different characteristics of IFPB and IFCB. IFPB calibration can be divided into constant and real-time IFPB calibrations to meet the different positioning requirements. Results show that constant IFPB rate has only 2 mm difference from the mean value of real-time IFPB rate. The IFPB rate and IFCB estimated by this algorithm have excellent stability, and the change in reference satellite cannot affect the results of IFPB rate and the stability of IFCB. The centimetre-level positioning results can be obtained using two calibration methods, and the positioning results with real-time calibration method are 10%–20% better than those with the constant calibration method. Under satellite-deprived environments, the improvements of multi-GNSS positioning accuracy with constant inter-frequency bias calibration gradually increase as the satellite cut-off elevation angle increases compared with GPS/BDS, which can reach up to 0·9 cm in the vertical direction.
The unacknowledged Panayotis Stamatakis and his invaluable contribution to the understanding of Grave Circle A at Mycenae
Authors: Konstantinidi-Syvridi Eleni , Paschalidis Constantinos
Published: December 2019

The legendary excavation of Mycenae’s Grave Circle A in 1876 resulted in the celebration of Heinrich and Sophia Schliemann as the sole excavators. The crucial role of Panayotis Stamatakis, the systematic and perceptive archaeologist who was assigned to supervise the excavation as a representative of the Greek state, was – for a long time – largely unacknowledged. Stamatakis’ invaluable manuscripts, where he recorded one by one all the finds and their locations, attributing them to specific burials, went missing for more than a century. Now part of the Archive of the National Archaeological Museum in Athens, these manuscripts shed new light on the taphonomy and burial customs of Grave Circle A, revealing the biographies of the first Mycenaean rulers.
Excavation and documentation of the Rhodian countryside and Dodecanese islands in the first millennium BC
Authors: Salmon Nicholas
Published: December 2019

This contribution offers an overview of recent fieldwork and museum-based projects focused on the Rhodian countryside and Dodecanese islands. The excavations conducted by the Ephorate of Antiquities of the Dodecanese over the past two decades, paired with the study of Rhodian collections in the Louvre and British Museum, among other museums, have developed and promoted the archaeological record of the region. The Kymissala Archaeological Research Project led by the University of the Aegean and a collaborative doctoral project investigating the British Museum’s collections from Kamiros each demonstrate the potential of revisiting historic excavations through topographical surveys and archival documentation.
The quality, understandability, readability, and popularity of online educational materials for heart murmur
Authors: Arslan Derya , Sami Tutar Mahmut , Kozanhan Betul , Bagci Zafer
Published: December 2019

Objective:

Murmurs are abnormal audible heart sounds produced by turbulent blood flow. Therefore, murmurs in a child may be a source of anxiety for family members. Families often use online materials to explore possible reasons for these murmurs, given the accessibility of information on the Internet. In this study, we evaluated the quality, understandability, readability, and popularity of online materials about heart murmur.

Methods:

An Internet search was performed for “heart murmur” using the Google search engine. The global quality score (on a scale of 1 to 5, corresponding to poor to excellent quality) and Health on the Net code were used to measure the quality of information presented. The understandability of the web pages identified was measured using the Patient Education Materials Assessment Tool (score range from 0 to 100%, scores below 70% reflect poor performance). The readability of each web pages was assessed using four validated indices: the Flesch Reading Ease Score, the Flesch–Kincaid Grade Level, the Gunning Frequency of Gobbledygook, and the Simple Measure of Gobbledygook. The ALEXA traffic tool was used to reference domains’ popularity and visibility.

Results:

We identified 230 English-language patient educational materials that discussed heart murmur. After exclusion, a total of 86 web pages were evaluated for this study. The average global quality score was 4.34 (SD = 0.71; range from 3 to 5) indicating that the quality of information of most websites was good. Only 14 (16.3%) websites had Health on the Net certification. The mean understandability score for all Internet-based patient educational materials was 74.6% (SD = 12.8%; range from 31.2 to 93.7%). A score suggesting these Internet-based patient educational materials were “easy to understand”. The mean readability levels of all patient educational materials were higher than the recommended sixth-grade reading level, according to all indices applied. This means that the level of readability is difficult. The average grade level for all web pages was 10.4 ± 1.65 (range from 7.53 to 14.13). The Flesch–Kincaid Grade level was 10 ± 1.81, the Gunning Frequency of Gobbledygook level was 12.1 ± 1.85, and the Simple Measure of Gobbledygook level was 9.1 ± 1.38. The average Flesch Reading Ease Score was 55 ± 9.1 (range from 32.4 to 72.9).

Conclusion:

We demonstrated that web pages describing heart murmurs were understandable and high quality. However, the readability level of the websites was above the recommended sixth-grade reading level. Readability of written materials from online sources need to be improved. However, care must be taken to ensure that the information of web pages is of a high quality and understandable.
Serbian Twin Registry
Authors: Smederevac Snežana , Mitrović Dušanka , Sadiković Selka , Milovanović Ilija , Branovački Bojan , Dinić Bojana M. , Nikolašević Željka , Kodžopeljić Jasmina , Bugarski Ignjatović Vojislava , Semnic Marija , Vujanić Stankov Tijana , Vučinić Nataša , Oljača Milan , Pajić Dejan , Stojadinović Aleksandra , Krstić Tatjana , Milutinović Aleksandra
Published: December 2019

The first twin study in Serbia began in 2011 as a part of the research project, ‘Psychological Foundations of Mental Health: Hereditary and Environmental Factors’. At the same time, the research team from the Faculty of Philosophy and Faculty of Medicine in Novi Sad established the first Serbian twin registry. The registry is intended primarily for the purpose of the research in behavioral genetics, as well as potential future studies in human genetics. It includes information on 1658 volunteers, including twin-pairs, their parent and siblings. The behavioral genetic study of adult twins has been focused on the hereditary and environmental sources of variance of different psychological characteristics, such as personality traits, cognitive abilities, executive functions and aggression, as well as some anthropometric measures and aspects of mental and physical health. Certain molecular genetic analyses have also been performed. The research team is currently starting the longitudinal twin study of children, which will be focused on different indicators of emotional, cognitive and physical development.
Observed Subdecadal Variations of European Summer Temperatures
Authors: Müller W.A. , Borchert L. , Ghosh R.
Published: December 2019

We identify subdecadal variations in European summer temperatures in coupled and uncoupled century‐long reanalyses. Spectral analyses reveal significant peaks at 5–10 years in the midtwentieth century. The subdecadal variations show substantial amplitudes of ~1–1.5 °C, associated with extremely warm summers during their positive phases. We use forced ocean model experiments and show that the European summer temperature variations are associated with the subdecadal coupled North Atlantic climate system. A positive winter NAO‐like forcing is associated with changes in the ocean circulation and mass and heat convergence occurring 1–2 years prior to European summer temperature rise. Ocean heat content and sea surface temperature increase in the subtropical North Atlantic. The atmospheric response is barotropic and induces wave activity fluxes toward the European continent, modulation of the jet positions, and blocking frequency. The atmospheric response establishes a pathway connecting the subdecadal coupled North Atlantic climate system to European summer temperature.
Solar Wind Deflection in the Foreshock: Model‐Data Comparison
Authors: Gutynska O. , Omidi N. , Sibeck D. G. , Nemecek Z. , Safrankova J. , Lynnyk A.
Published: December 2019

We present the results of a 2.5‐D (2‐D in space and 3‐D in currents and electromagnetic fields) electromagnetic hybrid simulation run for foreshock wave activity during an interval of a radial interplanetary magnetic field (IMF). The simulation predicts nonlinear waves and foreshock expansion in connection with the development of cavitons and the formation of the foreshock compressional boundary. Fluctuations of the ion velocity
component are associated with wave fronts corresponding to small wave normal angles and oscillate around 0 in the far upstream region. At distances closer to the bow shock, these fluctuations become asymmetric with duskward flow perturbations dominating at dusk and dawnward flow perturbations at dawn. We compare this simulation with THEMIS observations of large‐amplitude quasi‐monochromatic variations of ion velocity in the foreshock under radial IMF. A case study of simultaneous dual observations of unipolar
fluctuations is followed by a statistical study of similar solar wind deflections in the foreshock. The observations that confirm the simulation predictions indicate that, in addition to ion heating, ultralow‐frequency waves participate in the magnetosheath‐like plasma flow deflection in the foreshock during periods of small IMF cone angles.
Quasiperiodic Saturn Auroral Hiss Observed During a Cassini Proximal Orbit
Authors: Menietti J. D. , Palmaerts B. , Zahlava J. , Averkamp T. F. , Groene J. B. , Kurth W. S.
Published: December 2019

Saturn auroral hiss is intense whistler mode emission similar in morphology to terrestrial auroral hiss, and is observed at high latitude very often in quasiperiodic episodes with a period of approximately 1 hr. Bader et al. (2019) report auroral pulsations that may be due to duskside magnetodisk reconnection. The source of the 1‐hr period is not definitively known but has been purported to be due to second harmonic Alfven waves standing along near planet magnetic field lines (Yates et al., 2016). Observations of auroral hiss at high latitude along Cassini proximal orbits are often excellent, and we have focused on an event for which we have concurrent ultraviolet auroral images as well as electron flux data. A series of repeating auroral hiss episodes is observed to initiate near the magnetic field line that traverses a Saturn kilometric radiation source region in each hemisphere, with periodic episodes of hiss recurring at higher L‐shells. Magnetic field lines centered on individual hiss episodes have auroral footprints that lie near and within a region of intense auroral ultraviolet emissions. These observations have a parallel in terrestrial return current electron beam‐generated auroral hiss seen near magnetic field lines supporting auroral kilometric radiation source regions. Recent findings link periodic plasma injections with Saturn reconnection sites observed preferentially on the duskside. These injections may spawn Saturn kilometric radiation source regions and periodic auroral hiss emission in nearby return current regions.
Equatorial Magnetosonic Waves: Do Nonlinear Interactions Play a Role in Their Evolution?
Authors: Walker S. N. , Balikhin M. A. , Yearby K. H. , Aryan H.
Published: December 2019

The occurrence of nonlinear interactions between discrete wave frequencies has been shown to play a significant role in the propagation and evolution of some plasma wave modes. In this paper we take advantage of closely spaced Cluster measurements to investigate the possibility nonlinear interactions occurring between the discrete emissions that are observed in a region of Equatorial Magnetosonic Wave generation. Based on transfer function analysis, it is shown that the role of nonlinear interactions plays a negligible role in the wave evolution as the emissions propagate from one satellite to the other. A bicoherence analysis of the individual signals also fails to find the existence of nonlinear interactions in the evolution of equatorial magnetosonic waves.
Magnesium and Lithium Isotopic Evidence for a Remnant Oceanic Slab Beneath Central Tibet
Authors: Tian Heng‐Ci , Teng Fang‐Zhen , Hou Zeng‐Qian , Tian Shi‐Hong , Yang Wei , Chen Xin‐Yang , Song Yu‐Cai
Published: December 2019

The voluminous Eocene postcollisional potassium‐rich magmatism in central Tibet is crucial in understanding the deep geodynamic process and related uplift history of the Tibetan Plateau. Here, we present an integrated study of whole‐rock Sr‐Nd‐Mg‐Li isotopes and zircon U‐Pb ages on two suites of lavas from Duocaima and Nariniya in the northern Qiangtang block. The Duocaima high‐K calc‐alkaline trachyandesites were formed at ~35 Ma and are enriched in light rare earth elements and large ion lithophile elements. They have slightly lower δ26Mg (−0.41‰ to −0.33‰) and δ7Li (+1.0‰ to +2.6‰) relative to the mantle. Together with their high 87Sr/86Sr(i) (0.7066–0.7067) and low ɛNd(t) (−1.96 to −1.61) values, they most likely originated from the lithospheric mantle previously metasomatized by melts from subducted carbonate‐bearing sediments. Model calculation suggests that ~7–11% sediments were added into the mantle via melting. By contrast, the contemporaneous Nariniya adakitic trachytes have higher δ26Mg (−0.13‰ to −0.02‰) and δ7Li (+3.3‰ to +5.4‰) values, as well as higher Pb/Ce and Ba/La ratios, which most likely resulted from the thickened lower crust that was metasomatized by fluids released from downgoing Songpan‐Ganzi oceanic slab. Our combined isotopic study suggests the existence of a remnant oceanic slab beneath central Tibetan Plateau during the late Eocene and implies that the initial uplift of central Tibet was before ~38 Ma.
Enhancement of Atmospheric Stability by Anomalous Elevated Aerosols During Winter in China
Authors: Lu Xin , Mao Feiyue , Pan Zengxin , Gong Wei , Zhu Yannian , Yang Jie
Published: December 2019

Changes in anthropogenic aerosols with a low level of scientific cognition affect the atmospheric environment and radiative balance of the Earth. We determine anomalous elevated aerosol layers (EALs) and their source and effects on regional atmospheric stability in China by using 2007–2016 data from multiple satellite observations and models. Anomalous EALs occur during winter over Eastern China, and they are mainly distributed above 2 km and can even spread up to 4–5 km despite the disadvantageous atmospheric convection conditions during winter. Anthropogenic pollutants that mainly originate from local‐range transport with enhanced atmospheric updraft play a dominant role (about 60%) in this situation. Furthermore, anomalous EALs result in considerable warming in the upper atmosphere (+0.32 K/day) and cooling on the surface (−0.04 K/day) over Eastern China. They decrease surface incident radiation by an average of 3.4 W/m2. The opposite heating rate change in the upper atmosphere and surface caused by anomalous EALs consequently increases the low atmospheric stability by an average of 0.39 K, and this increment weakens the dissipation of atmospheric dispersion. Related studies have indicated that increased atmospheric stability can severely exacerbate atmospheric pollution near the surface.
Laboratory and Numerical Investigation of Saltwater Intrusion Processes in a Circular Island Aquifer
Authors: Memari Sama S. , Bedekar Vivek S. , Clement T. Prabhakar
Published: December 2019

A novel radial flow experimental system to study saltwater intrusion processes in an island aquifer is presented. The study investigated steady state and transient scenarios involving advancing and receding saltwater wedges in a circular island. The experimental results were simulated using the density‐coupled version of the MODFLOW‐USG code. The experimental data along with the model simulation results are employed to develop a new radial benchmark problem for testing density‐coupled models used for simulating saltwater intrusion processes. The experimental data for transient changes in toe position and freshwater storage level indicated an asymmetric pattern where the intrusion time scale is greater than the recession time scale. Numerical experiments were completed to further investigate this asymmetric effect and to intercompare the associated transient transport processes in circular and linear strip islands. We also analyzed the sensitivity of island geometry in controlling the freshwater storage levels under different recharge conditions. Modeling results show that for similar‐sized systems, circular islands are more efficient in storing freshwater than linear strip islands.
Characterization and printability of Sodium alginate -Gelatin hydrogel for bioprinting NSCLC co-culture
Authors: Mondal Arindam , Gebeyehu Aragaw , Miranda Mariza , Bahadur Divya , Patel Nilkumar , Ramakrishnan Subhramanian , Rishi Arun K. , Singh Mandip
Published: December 2019

3D bioprinting improves orientation of in vitro tumor models by offering layer by layer positioning of cancer cells and cancer associated fibroblasts (CAFs) which can replicate tumor microenvironment. Aim of this study was to develop a sodium alginate -gelatin (SA-GL) hydrogel by optimizing rheological parameters to print non-small cell lung cancer (NSCLC) patient derived xenograft (PDX) cells and lung CAFs co-cultures. SA-GL hydrogels were prepared, and rheological properties were evaluated. Both the cells were mixed with the hydrogel and printed using INKREDIBLE bioprinter. Hydrogels prepared with 3.25% and 3.5% (w/v) SA and 4% (w/v) GL showed higher printability and cell viability. A significant decline in viscosity with shear rate was observed in these hydrogels suggesting the shear thinning property of hydrogels. Spheroid size distribution after 15 days was in the diameter range of 50–1100 µm. Up-regulation of vimentin, α-SMA and loss of E-cadherin in co-culture spheroids confirmed cellular crosstalk. This study demonstrates that rheological optimization of SA-GL hydrogel enhances printability and viability of NSCLC PDX and CAF co-culture which allows 3D co-culture spheroid formation within the printed scaffold. Therefore, this model can be used for studying high throughput drug screening and other pre-clinical applications.
Quantitative MRI Biomarkers of Stereotactic Radiotherapy Outcome in Brain Metastasis
Authors: Karami Elham , Soliman Hany , Ruschin Mark , Sahgal Arjun , Myrehaug Sten , Tseng Chia-Lin , Czarnota Gregory J. , Jabehdar-Maralani Pejman , Chugh Brige , Lau Angus , Stanisz Greg J. , Sadeghi-Naini Ali
Published: December 2019

About 20–40% of cancer patients develop brain metastases, causing significant morbidity and mortality. Stereotactic radiation treatment is an established option that delivers high dose radiation to the target while sparing the surrounding normal tissue. However, up to 20% of metastatic brain tumours progress despite stereotactic treatment, and it can take months before it is evident on follow-up imaging. An early predictor of radiation therapy outcome in terms of tumour local failure (LF) is crucial, and can facilitate treatment adjustments or allow for early salvage treatment. In this study, an MR-based radiomics framework was proposed to derive and investigate quantitative MRI (qMRI) biomarkers for the outcome of LF in brain metastasis patients treated with hypo-fractionated stereotactic radiation therapy (SRT). The qMRI biomarkers were constructed through a multi-step feature extraction/reduction/selection framework using the conventional MR imaging data acquired from 100 patients (133 lesions), and were applied in conjunction with machine learning techniques for outcome prediction and risk assessment. The results indicated that the majority of the features in the optimal qMRI biomarkers characterize the heterogeneity in the surrounding regions of tumour including edema and tumour/lesion margins. The optimal qMRI biomarker consisted of five features that predict the outcome of LF with an area under the curve (AUC) of 0.79, and a cross-validated sensitivity and specificity of 81% and 79%, respectively. The Kaplan-Meier analyses showed a statistically significant difference in local control (p-value < 0.0001) and overall survival (p = 0.01). Findings from this study are a step towards using qMRI for early prediction of local failure in brain metastasis patients treated with SRT. This may facilitate early adjustments in treatment, such as surgical resection or salvage radiation, that can potentially improve treatment outcomes. Investigations on larger cohorts of patients are, however, required for further validation of the technique.
Experimental arthritis and Porphyromonas gingivalis administration synergistically decrease bone regeneration in femoral cortical defects
Authors: Okumura Go , Kondo Naoki , Sato Keisuke , Yamazaki Kazuhisa , Ohshima Hayato , Kawashima Hiroyuki , Ogose Akira , Endo Naoto
Published: December 2019

Porphyromonas gingivalis infection can lead to periodontitis and dysbiosis, which are known risk factors for rheumatoid arthritis (RA). We investigated whether P. gingivalis administration affected bone regeneration in mice with or without arthritis. We administered P. gingivalis to male DBA/1 J mice that were or were not sensitised to type II collagen-induced arthritis (CIA). All mice underwent drilling of bilateral femurs. We histologically evaluated new bone regeneration (bone volume of the defect [BVd]/tissue volume of the defect [TVd]) using micro-computed tomography (micro-CT), osteoclast number/bone area, and active osteoblast surface/bone surface (Ob.S/BS). We measured serum cytokine levels and bone mineral density of the proximal tibia using micro-CT. CIA resulted in significantly reduced bone regeneration (BVd/TVd) at all time-points, whereas P. gingivalis administration showed similar effects at 2 weeks postoperatively. CIA resulted in higher osteoclast number/bone area and lower Ob.S/BS at 2 and 3 weeks postoperatively, respectively. However, P. gingivalis administration resulted in lower Ob.S/BS only at 2 weeks postoperatively. During later-stage bone regeneration, CIA and P. gingivalis administration synergistically decreased BVd/TVd, increased serum tumour necrosis factor-α, and resulted in the lowest bone mineral density. Therefore, RA and dysbiosis could be risk factors for prolonged fracture healing.
A CRISPR-Cas9 System for Genome Editing of Fusarium proliferatum
Authors: Ferrara Massimo , Haidukowski Miriam , Logrieco Antonio F. , Leslie John F. , Mulè Giuseppina
Published: December 2019

Fusarium proliferatumcauses diverse diseases of many economically important plants. The fungus produces several mycotoxins of which the fumonisins are the most toxic. Currently, deletion of key genes for mycotoxin biosynthesis is a laborious and time-consuming procedure. We developed a novel CRISPR/Cas9-based genome-editing tool for the direct delivery of preassembled Cas9 ribonucleoproteins into protoplasts ofF. proliferatum. Our CRISPR–Cas9 system couples a site-specific double-strand DNA break mediated by two Cas9 ribonucleoproteins with microhomology recombination requiring only 50-bp regions flanking the target gene. This system reduces the risk of off-target mutations and minimizes the risk of altering any gene adjacent to the target region. We used this tool to delete a polyketide synthase gene (FUM1) required for fumonisin biosynthesis. The mutants generated are no longer able to produce fumonisins, confirming the key role ofFUM1in fumonisin biosynthesis. Our CRISPR-Cas9 system is an important new tool for genetic studies ofFusarium.
Dissecting multi drug resistance in head and neck cancer cells using multicellular tumor spheroids
Authors: Azharuddin Mohammad , Roberg Karin , Dhara Ashis Kumar , Jain Mayur Vilas , Darcy Padraig , Hinkula Jorma , Slater Nigel K. H. , Patra Hirak K.
Published: December 2019

One of the hallmarks of cancers is their ability to develop resistance against therapeutic agents. Therefore, developing effective in vitro strategies to identify drug resistance remains of paramount importance for successful treatment. One of the ways cancer cells achieve drug resistance is through the expression of efflux pumps that actively pump drugs out of the cells. To date, several studies have investigated the potential of using 3-dimensional (3D) multicellular tumor spheroids (MCSs) to assess drug resistance; however, a unified system that uses MCSs to differentiate between multi drug resistance (MDR) and non-MDR cells does not yet exist. In the present report we describe MCSs obtained from post-diagnosed, pre-treated patient-derived (PTPD) cell lines from head and neck squamous cancer cells (HNSCC) that often develop resistance to therapy. We employed an integrated approach combining response to clinical drugs and screening cytotoxicity, monitoring real-time drug uptake, and assessing transporter activity using flow cytometry in the presence and absence of their respective specific inhibitors. The report shows a comparative response to MDR, drug efflux capability and reactive oxygen species (ROS) activity to assess the resistance profile of PTPD MCSs and two-dimensional (2D) monolayer cultures of the same set of cell lines. We show that MCSs provide a robust and reliable in vitro model to evaluate clinical relevance. Our proposed strategy can also be clinically applicable for profiling drug resistance in cancers with unknown resistance profiles, which consequently can indicate benefit from downstream therapy.
A natural polymorphism in Zika virus NS2A protein responsible of virulence in mice
Authors: Ávila-Pérez Gines , Nogales Aitor , Park Jun-Gyu , Márquez-Jurado Silvia , Iborra Francisco J. , Almazan Fernando , Martínez-Sobrido Luis
Published: December 2019

Zika virus (ZIKV) infection is currently one of the major concerns in human public health due to its association with neurological disorders. Intensive effort has been implemented for the treatment of ZIKV, however there are not currently approved vaccines or antivirals available to combat ZIKV infection. In this sense, the identification of virulence factors associated with changes in ZIKV virulence could help to develop safe and effective countermeasures to treat ZIKV or to prevent future outbreaks. Here, we have compared the virulence of two related ZIKV strains from the recent outbreak in Brazil (2015), Rio Grande do Norte Natal (RGN) and Paraiba. In spite of both viruses being identified in the same period of time and region, significant differences in virulence and replication were observed using a validated mouse model of ZIKV infection. While ZIKV-RGN has a 50% mouse lethal dose (MLD50) of ~105 focus forming units (FFUs), ZIKV-Paraiba infection resulted in 100% of lethality with less than 10 FFUs. Combining deep-sequencing analysis and our previously described infectious ZIKV-RGN cDNA clone, we identified a natural polymorphism in the non-structural protein 2 A (NS2A) that increase the virulence of ZIKV. Moreover, results demonstrate that the single amino acid alanine to valine substitution at position 117 (A117V) in the NS2A was sufficient to convert the attenuated rZIKV-RGN in a virulent Paraiba-like virus (MLD50 < 10 FFU). The mechanism of action was also evaluated and data indicate that substitution A117V in ZIKV NS2A protein reduces host innate immune responses and viral-induced apoptosis in vitro. Therefore, amino acid substitution A117V in ZIKV NS2A could be used as a genetic risk-assessment marker for future ZIKV outbreaks.
Prognostic factors and outcome of adult allogeneic hematopoietic stem cell transplantation patients admitted to intensive care unit during transplant hospitalization
Authors: Michel Christian S. , Teschner Daniel , Schmidtmann Irene , Theobald Matthias , Hauptrock Beate , Wagner-Drouet Eva M. , Radsak Markus P.
Published: December 2019

Patients undergoing allogeneic hematopoietic stem cell transplantation have a high morbidity and mortality, especially after admission to intensive care unit (ICU) during peri-transplant period. The objective of this study was to identify new clinical and biological parameters and validate prognostic scores associated with ICU, short-and long-term survival. Significant differences between ICU survivors and ICU non-survivors for the clinical parameters invasive mechanical ventilation, urine output, heart rate, mean arterial pressure, and amount of vasopressors have been measured. Among prognostic scores (SOFA, SAPSII, PICAT, APACHE II, APACHE IV) assessing severity of disease and predicting outcome of critically ill patients on ICU, the APACHE II score has shown most significant difference (p = 0.002) and the highest discriminative power (area under the ROC curve (AUC) 0.74). An elevated level of lactate at day of admission was associated with poor survival on ICU and the most significant independent parameter (p < 0.001). In our cohort kidney damage with low urine output has a highly relevant impact on ICU, short- and long-term overall survival. The APACHE II score was superior predicting ICU mortality compared to all other tested prognostic scores for patients on ICU during peri-transplant period.
Differentiating external zeitgeber impact on peripheral circadian clock resetting
Authors: Heyde Isabel , Oster Henrik
Published: December 2019

Circadian clocks regulate physiological functions, including energy metabolism, along the 24-hour day cycle. The mammalian clock system is organized in a hierarchical manner with a coordinating pacemaker residing in the hypothalamic suprachiasmatic nucleus (SCN). The SCN clock is reset primarily by the external light-dark cycle while other zeitgebers such as the timing of food intake are potent synchronizers of many peripheral tissue clocks. Under conflicting zeitgeber conditions, e.g. during shift work, phase synchrony across the clock network is disrupted promoting the development of metabolic disorders. We established a zeitgeber desynchrony (ZD) paradigm to quantify the differential contributions of the two main zeitgebers, light and food, to the resetting of specific tissue clocks and the effect on metabolic homeostasis in mice. Under 28-hour light-dark and 24-hour feeding-fasting conditions SCN and peripheral clock, as well as activity and hormonal rhythms showed specific periodicities aligning in-between those of the two zeitgebers. During ZD, metabolic homeostasis was cyclic with mice gaining weight under synchronous and losing weight under conflicting zeitgeber conditions. In summary, our study establishes an experimental paradigm to compare zeitgeber input in vivo and study the physiological consequences of chronodisruption.
Restoring colistin sensitivity in colistin-resistant E. coli: Combinatorial use of MarR inhibitor with efflux pump inhibitor
Authors: Sundaramoorthy Niranjana Sri , Suresh Pavithira , Selva Ganesan Subramaniapillai , GaneshPrasad ArunKumar , Nagarajan Saisubramanian
Published: December 2019

Antibiotics like colistin are the last resort to deal with infections by carbapenem-resistant Enterobacteriaceae (CREB). Resistance to colistin severely restricts therapeutic options. To tackle this dire situation, urgent measures to restore colistin sensitivity are needed. In this study, whole-genome sequencing of colistin-resistant E. coli strain was performed and the genome analysis revealed that the strain belonged to the sequence type ST405. Multiple mutations were observed in genes implicated in colistin resistance, especially those related to the L-Ara-4-N pathway but mgrB was unmutated and mcr1-9 genes were missing. MarR inhibitor salicylate was used to re-sensitize this strain to colistin, which increased the negative charge on the cell surface especially in colistin resistant E. coli (U3790 strain) and thereby facilitated a decrease in colistin MIC by 8 fold. It is indeed well known that MarR inhibition by salicylate triggers the expression of AcrAB efflux pumps through MarA. So, in order to fully restore colistin sensitivity, a potent efflux pump inhibitor (BC1), identified earlier by this group was employed. The combination of colistin with both salicylate and BC1 caused a remarkable 6 log reduction in cell counts of U3790 in time-kill assay. Infection of muscle tissue of zebrafish with U3790 followed by various treatments showed that the combination of colistin + salicylate + BC1 was highly effective in reducing bioburden in infected muscle tissue by 4 log fold. Thus, our study shows that a combination of MarR inhibitor to enhance colistin binding and efflux pump inhibitor to reduce colistin extrusion was highly effective in restoring colistin sensitivity in colistin-resistant clinical isolate of E. coli in vitro and in vivo.
Effects of cold exposure revealed by global transcriptomic analysis in ferret peripheral blood mononuclear cells
Authors: Reynés Bàrbara , van Schothorst Evert M. , Keijer Jaap , Palou Andreu , Oliver Paula
Published: December 2019

Animal studies, mostly performed in rodents, show the beneficial anti-obesity effects of cold studies. This is due to thermogenic activation of brown adipose tissue (BAT), a tissue also recently discovered in adult humans. Studies in humans, however, are hampered by the accessibility of most tissues. In contrast, peripheral blood mononuclear cells (PBMC) are accessible and share the expression profile of different sets of genes with other tissues, including those that reflect metabolic responses. Ferrets are an animal model physiologically closer to humans than rodents. Here, we investigated the effects on ferrets of one-week acclimation to 4 °C by analysing the PBMC transcriptome. Cold exposure deeply affected PBMC gene expression, producing a widespread down-regulation of genes involved in different biological pathways (cell cycle, gene expression regulation/protein synthesis, immune response, signal transduction, and genes related to extracellular matrix/cytoskeleton), while thermogenic and glycogenolysis-related processes were increased. Results obtained in PBMC reflected those of adipose tissue, but hardly those of the liver. Our study, using ferret as a model, reinforce PBMC usefulness as sentinel biological material for cold-exposure studies in order to deepen our understanding of the general and specific pathways affected by cold acclimation. This is relevant for future development of therapies to be used clinically.
Place of death trends among patients with dementia in Japan: a population-based observational study
Authors: Koyama Toshihiro , Sasaki Misato , Hagiya Hideharu , Zamami Yoshito , Funahashi Tomoko , Ohshima Ayako , Tatebe Yasuhisa , Mikami Naoko , Shinomiya Kazuaki , Kitamura Yoshihisa , Sendo Toshiaki , Hinotsu Shiro , Kano Mitsunobu R.
Published: December 2019

Dementia is a major public health concern in ageing societies. Although the population of Japan is among the most aged worldwide, long-term trends in the place of death (PoD) among patients with dementia is unknown. In this Japanese nationwide observational study, we analysed trends in PoD using the data of patients with dementia who were aged ≥65 years and died during 1999–2016. Trends in the crude death rates and PoD frequencies were analysed using the Joinpoint regression model. Changes in these trends were assessed using the Joinpoint regression analysis in which significant change points, the annual percentage change (APC) and average APCs (AAPC) in hospitals, homes, or nursing homes were estimated. During 1999–2016, the number of deaths among patients with dementia increased from 3,235 to 23,757 (total: 182,000). A trend analysis revealed increased mortality rates, with an AAPC of 8.2% among men and 9.3% among women. Most patients with dementia died in the hospital, although the prevalence of hospital deaths decreased (AAPC: -1.0%). Moreover, the prevalence of nursing home deaths increased (AAPC: 5.6%), whereas the prevalence of home deaths decreased (AAPC: -5.8%). These findings support a reconsideration of the end-of-life care provided to patients with dementia.
Genetic risk score for risk prediction of diabetic nephropathy in Han Chinese type 2 diabetes patients
Authors: Liao Li-Na , Li Tsai-Chung , Li Chia-Ing , Liu Chiu-Shong , Lin Wen-Yuan , Lin Chih-Hsueh , Yang Chuan-Wei , Chen Ching-Chu , Chang Chiz-Tzung , Yang Ya-Fei , Liu Yao-Lung , Kuo Huey-Liang , Tsai Fuu-Jen , Lin Cheng-Chieh
Published: December 2019

We evaluated whether genetic information could offer improvement on risk prediction of diabetic nephropathy (DN) while adding susceptibility variants into a risk prediction model with conventional risk factors in Han Chinese type 2 diabetes patients. A total of 995 (including 246 DN cases) and 519 (including 179 DN cases) type 2 diabetes patients were included in derivation and validation sets, respectively. A genetic risk score (GRS) was constructed with DN susceptibility variants based on findings of our previous genome-wide association study. In derivation set, areas under the receiver operating characteristics (AUROC) curve (95% CI) for model with clinical risk factors only, model with GRS only, and model with clinical risk factors and GRS were 0.75 (0.72–0.78), 0.64 (0.60–0.68), and 0.78 (0.75–0.81), respectively. In external validation sample, AUROC for model combining conventional risk factors and GRS was 0.70 (0.65–0.74). Additionally, the net reclassification improvement was 9.98% (P = 0.001) when the GRS was added to the prediction model of a set of clinical risk factors. This prediction model enabled us to confirm the importance of GRS combined with clinical factors in predicting the risk of DN and enhanced identification of high-risk individuals for appropriate management of DN for intervention.
The neurocognitive gains of diagnostic reasoning training using simulated interactive veterinary cases
Authors: Nassar Maaly
Published: December 2019

The present longitudinal study ascertained training-associated transformations in the neural underpinnings of diagnostic reasoning, using a simulation game named “Equine Virtual Farm” (EVF). Twenty participants underwent structural, EVF/task-based and resting-state MRI and diffusion tensor imaging (DTI) before and after completing their training on diagnosing simulated veterinary cases. Comparing playing veterinarian versus seeing a colorful image across training sessions revealed the transition of brain activity from scientific creativity regions pre-training (left middle frontal and temporal gyrus) to insight problem-solving regions post-training (right cerebellum, middle cingulate and medial superior gyrus and left postcentral gyrus). Further, applying linear mixed-effects modelling on graph centrality metrics revealed the central roles of the creative semantic (inferior frontal, middle frontal and angular gyrus and parahippocampus) and reward systems (orbital gyrus, nucleus accumbens and putamen) in driving pre-training diagnostic reasoning; whereas, regions implicated in inductive reasoning (superior temporal and medial postcentral gyrus and parahippocampus) were the main post-training hubs. Lastly, resting-state and DTI analysis revealed post-training effects within the occipitotemporal semantic processing region. Altogether, these results suggest that simulation-based training transforms diagnostic reasoning in novices from regions implicated in creative semantic processing to regions implicated in improvised rule-based problem-solving.
Biological Carbon Recovery from Sugar Refinery Washing Water into Microalgal DHA: Medium Optimization and Stress Induction
Authors: Moon Myounghoon , Park Won-Kun , Suh William I. , Chang Yong Keun , Lee Bongsoo
Published: December 2019

Sugar refinery washing water (SRWW) contains abundant levels of carbon sources and lower levels of contaminants than other types of wastewater, which makes it ideal for heterotrophic cultivation of microalgae. Here, carbon sources in SRWW were utilized for conversion into the form of value-added docosahexaenoic acid (DHA) usingAurantiochytriumsp. KRS101. Since SRWW is not a defined medium, serial optimizations were performed to maximize the biomass, lipid, and DHA yields by adjusting the nutrient (carbon, nitrogen, and phosphorus) concentrations as well as the application of salt stress. Optimum growth performance was achieved with 30% dilution of SRWW containing a total organic carbon of 95,488 mg L−1. Increasing the nutrient level in the medium by supplementation of 9 g L−1KH2PO4and 20 g L−1yeast extract further improved the biomass yield by an additional 14%, albeit at the expense of a decrease in the lipid content. Maximum biomass, lipid, and DHA yields (22.9, 6.33, and 2.03 g L−1, respectively) were achieved when 35 g L−1sea salt was applied on a stationary phase for osmotic stress. These results demonstrate the potential of carbon-rich sugar refinery washing water for DHA production usingAurantiochytriumsp. KRS101 and proper cultivation strategy.
Evaluation of mitochondria in oocytes following γ-irradiation
Authors: Wang Qiaochu , Stringer Jessica M. , Liu Jun , Hutt Karla J.
Published: December 2019

Standard cytotoxic cancer treatments, such as radiation, can damage and deplete the supply of oocytes stored within the ovary, which predisposes females to infertility and premature menopause later in life. The mechanisms by which radiation induces oocyte damage have not been completely elucidated. The objective of this study was to determine if γ-irradiation changes mitochondrial characteristics in oocytes, possibly contributing to a reduction in oocyte number and quality. Immature oocytes were collected from postnatal day (PN) 9–11 C57Bl6 mice 3, 6 and 24 hours after 0.1 Gy γ-irradiation to monitor acute mitochondrial changes. Oocytes were classified as small (>20 µm) or growing (40–60 µm). Mitochondrial membrane potential was lost in 20% and 44% of small oocytes (~20 µm) at 3 and 6 hours after γ-irradiation, respectively, consistent with the induction of apoptosis. However, mitochondrial mass, distribution and membrane potential in the surviving small oocytes were similar to the non-irradiated controls at both time points. At 24 hours after γ-irradiation, all mitochondrial parameters analysed within immature oocytes were similar to untreated controls. Mitochondrial parameters within growing oocytes were also similar to untreated controls. When mice were superovulated more than 3 weeks after γ-irradiation, there was a significant reduction in the number of mature oocytes harvested compared to controls (Control 18 ± 1 vs 0.1 Gy 4 ± 1, n = 6/16 mice, p < 0.05). There was a slight reduction in mitochondrial mass in mature oocytes after γ-irradiation, though mitochondrial localization, mtDNA copy number and ATP levels were similar between groups. In summary, this study shows that γ-irradiation of pre-pubertal mice is associated with loss of mitochondrial membrane potential in a significant proportion of small immature oocytes and a reduction in the number of mature oocytes harvested from adult mice. Furthermore, these results suggest that immature oocytes that survive γ-irradiation and develop through to ovulation contain mitochondria with normal characteristics. Whether the oocytes that survive radiation and eventually undergo meiosis can support fertility remains to be determined.
Impact of spaceflight on the murine thymus and mitigation by exposure to artificial gravity during spaceflight
Authors: Horie Kenta , Kato Tamotsu , Kudo Takashi , Sasanuma Hiroki , Miyauchi Maki , Akiyama Nobuko , Miyao Takahisa , Seki Takao , Ishikawa Tatsuya , Takakura Yuki , Shirakawa Masaki , Shiba Dai , Hamada Michito , Jeon Hyojung , Yoshida Nobuaki , Inoue Jun-ichiro , Muratani Masafumi , Takahashi Satoru , Ohno Hiroshi , Akiyama Taishin
Published: December 2019

The environment experienced during spaceflight may impact the immune system and the thymus appears to undergo atrophy during spaceflight. However, molecular aspects of this thymic atrophy remain to be elucidated. In this study, we analysed the thymi of mice on board the international space station (ISS) for approximately 1 month. Thymic size was significantly reduced after spaceflight. Notably, exposure of mice to 1 × gusing centrifugation cages in the ISS significantly mitigated the reduction in thymic size. Although spaceflight caused thymic atrophy, the global thymic structure was not largely changed. However, RNA sequencing analysis of the thymus showed significantly reduced expression of cell cycle-regulating genes in two independent spaceflight samples. These reductions were partially countered by 1 × gexposure during the space flights. Thus, our data suggest that spaceflight leads to reduced proliferation of thymic cells, thereby reducing the size of the thymus, and exposure to 1 × gmight alleviate the impairment of thymus homeostasis induced by spaceflight.
Genomic and functional gene studies suggest a key role of beta-carotene oxygenase 1 like (bco1l) gene in salmon flesh color
Authors: Helgeland Hanna , Sodeland Marte , Zoric Nina , Torgersen Jacob Seilø , Grammes Fabian , von Lintig Johannes , Moen Thomas , Kjøglum Sissel , Lien Sigbjørn , Våge Dag Inge
Published: December 2019

Red coloration of muscle tissue (flesh) is a unique trait in several salmonid genera, including Atlantic salmon. The color results from dietary carotenoids deposited in the flesh, whereas the color intensity is affected both by diet and genetic components. Herein we report on a genome-wide association study (GWAS) to identify genetic variation underlying this trait. Two SNPs on ssa26 showed strong associations to the flesh color in salmon. Two genes known to be involved in carotenoid metabolism were located in this QTL- region: beta-carotene oxygenase 1 (bco1) and beta-carotene oxygenase 1 like (bco1l). To determine whether flesh color variation is caused by one, or both, of these genes, functional studies were carried out including mRNA and protein expression in fish with red and pale flesh color. The catalytic abilities of these two genes were also tested with different carotenoids. Our results suggest bco1l to be the most likely gene to explain the flesh color variation observed in this population.