bluuebunny/crossref_metadata_2025 huggingface dataset:
Respiratory syncytial virus A genotype classification based on systematic intergenotypic and intragenotypic sequence analysis
Authors: Muñoz-Escalante Juan Carlos , Comas-García Andreu , Bernal-Silva Sofía , Robles-Espinoza Carla Daniela , Gómez-Leal Guillermo , Noyola Daniel E.
Published: December 2019

Respiratory syncytial virus (RSV), a leading cause of lower respiratory tract infections, is classified in two major groups (A and B) with multiple genotypes within them. Continuous changes in spatiotemporal distribution of RSV genotypes have been recorded since the identification of this virus. However, there are no established criteria for genotype definition, which affects the understanding of viral evolution, immunity, and development of vaccines. We conducted a phylogenetic analysis of 4,353 RSV-A G gene ectodomain sequences, and used 1,103 complete genome sequences to analyze the totallity of RSV-A genes. Intra- and intergenotype p-distance analysis and identification of molecular markers associated to specific genotypes were performed. Our results indicate that previously reported genotypes can be classified into nine distinct genotypes: GA1-GA7, SAA1, and NA1. We propose the analysis of the G gene ectodomain with a wide set of reference sequences of all genotypes for an accurate genotype identification.
Structure-Guided Design of a Fluorescent Probe for the Visualization of FtsZ in Clinically Important Gram-Positive and Gram-Negative Bacterial Pathogens
Authors: Ferrer-González Edgar , Fujita Junso , Yoshizawa Takuya , Nelson Julia M. , Pilch Alyssa J. , Hillman Elani , Ozawa Mayuki , Kuroda Natsuko , Al-Tameemi Hassan M. , Boyd Jeffrey M. , LaVoie Edmond J. , Matsumura Hiroyoshi , Pilch Daniel S.
Published: December 2019

Addressing the growing problem of antibiotic resistance requires the development of new drugs with novel antibacterial targets. FtsZ has been identified as an appealing new target for antibacterial agents. Here, we describe the structure-guided design of a new fluorescent probe (BOFP) in which a BODIPY fluorophore has been conjugated to an oxazole-benzamide FtsZ inhibitor. Crystallographic studies have enabled us to identify the optimal position for tethering the fluorophore that facilitates the high-affinity FtsZ binding of BOFP. Fluorescence anisotropy studies demonstrate that BOFP binds the FtsZ proteins from the Gram-positive pathogens Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium, Streptococcus pyogenes, Streptococcus agalactiae, and Streptococcus pneumoniae with Kd values of 0.6–4.6 µM. Significantly, BOFP binds the FtsZ proteins from the Gram-negative pathogens Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii with an even higher affinity (Kd = 0.2–0.8 µM). Fluorescence microscopy studies reveal that BOFP can effectively label FtsZ in all the above Gram-positive and Gram-negative pathogens. In addition, BOFP is effective at monitoring the impact of non-fluorescent inhibitors on FtsZ localization in these target pathogens. Viewed as a whole, our results highlight the utility of BOFP as a powerful tool for identifying new broad-spectrum FtsZ inhibitors and understanding their mechanisms of action.
Carrier Induced Hopping to Band Conduction in Pentacene
Authors: Rani Varsha , Kumar Pramod , Sharma Akanksha , Yadav Sarita , Singh Budhi , Ray Nirat , Ghosh Subhasis
Published: December 2019

Charge transport in organic thin films which are generally polycrystalline is typically limited by the localization of the carriers at lattice defects resulting in low carrier mobilities and carriers move from one state to another state by hopping. However, charge transport in organic semiconductors in their single crystalline phase is coherent due to band conduction and mobilities are not limited by disorder resulting in higher carrier mobility. So it is a challenge to enhance the carrier mobility in a thin film which is the preferred choice for all organic devices. Here, we show that it is possible to increase the carrier mobility in polycrystalline thin films by injecting sufficient carriers such that Fermi level can be moved into the region of high density in Gaussian density of states of molecular solids. When the hopping transport happens through the molecular energy levels whose density is low, mobility is decided by incoherent transport however, when the the hopping transport happens through the energy levels with high density, mobility is decided by coherent transport, as in band conduction. We present results highlighting the observation of both band-like and hopping conduction in polycrystalline organic thin films by varying the concentration of injected charge. More importantly the transition from hopping to band transport is reversible. The observed carrier mobilities in both the regimes match well with theoretical estimates of hopping mobility and band mobility determined from first principles density functional theory.
Critical role of CRAG, a splicing variant of centaurin-γ3/AGAP3, in ELK1-dependent SRF activation at PML bodies
Authors: Nagashima Shun , Takeda Keisuke , Shiiba Isshin , Higashi Mizuho , Fukuda Toshifumi , Tokuyama Takeshi , Matsushita Nobuko , Nagano Seiichi , Araki Toshiyuki , Kaneko Mari , Shioi Go , Inatome Ryoko , Yanagi Shigeru
Published: December 2019

CRMP-5-associated GTPase (CRAG), a short splicing variant of centaurin-γ3/AGAP3, is predominantly expressed in the developing brain. We previously demonstrated that CRAG, but not centaurin-γ3, translocates to the nucleus and activates the serum response factor (SRF)-c-Fos pathway in cultured neuronal cells. However, the physiological relevance of CRAG in vivo is unknown. Here, we found that CRAG/centaurin-γ3–knockout mice showed intensively suppressed kainic acid-induced c-fos expression in the hippocampus. Analyses of molecular mechanisms underlying CRAG-mediated SRF activation revealed that CRAG has an essential role in GTPase activity, interacts with ELK1 (a co-activator of SRF), and activates SRF in an ELK1-dependent manner. Furthermore, CRAG and ELK1 interact with promyelocytic leukaemia bodies through SUMO-interacting motifs, which is required for SRF activation. These results suggest that CRAG plays a critical role in ELK1-dependent SRF-c-fos activation at promyelocytic leukaemia bodies in the developing brain.
Simultaneous influence of sympathetic autonomic stress on Schlemm’s canal, intraocular pressure and ocular circulation
Authors: Chen Wei , Chen Zhiqi , Xiang Yan , Deng Chaohua , Zhang Hong , Wang Junming
Published: December 2019

This study aimed to investigate changes in Schlemm’s canal, intraocular pressure and ocular blood circulation following the activation of the sympathetic nervous system. Twenty healthy volunteers were enrolled in this study. The cold pressor test (CPT) was adopted. Cross-sectional area of Schlemm’s canal (SCAR), superficial and deep retinal vessel densities (s-RVD;d-RVD), pupil diameter (PD), intraocular pressure (IOP), mean ocular perfusion pressure (MOPP) and heart rate variability (HRV) were measured at three time-points: baseline (T0) and 5 min (T1) and 10 min (T2) after the CPT. After cold stimulation, LF/HF index (the ratio of low frenquency and high frenquency) increased significantly. IOP decreased from 16.9 ± 1.9 mmHg at baseline to 16.4 ± 2.7 mmHg at T1 and to 15.2 ± 2.7 mmHg at T2. The nasal cross-sectional area of SCAR (SCAR-n) increased from 6283.9 ± 2696.2 µm2 at baseline to 8392.9 ± 3258.7 µm2 at T1 and to 10422.0 ± 3643.8 µm2 at T2. The temporal cross-sectional area of SCAR (SCAR-t) increased from 6414.5 ± 2218.7 µm2 at baseline to 8610.8 ± 2317.1 µm2 at T1 and to 11544.0 ± 4129.2 µm2 at T2. The expansion of Schlemm’s canal was observed after the CPT might be caused by sympathetic nerve stimulation, subsequently leading to decreased IOP.
A novel index equivalent to the myocardial performance index for right ventricular functional assessment in children and adolescent patients
Authors: Hayabuchi Yasunobu , Homma Yukako , Kagami Shoji
Published: December 2019

The aims of the present study were to develop and check the utility and feasibility of a novel right ventricular (RV) functional index (RV angular velocity; RVω, s−1) derived from the angular velocity in harmonic oscillator kinematics obtained from the RV pressure waveform. We hypothesized that RVω reflects the myocardial performance index (MPI), which represents global RV function. A total of 132 consecutive patients, ranging in age from 3 months to 34 years with various cardiac diseases were included in this prospective study. RVω was defined as the difference between the peak derivative of pressure (dP/dt_max − dP/dt_min) divided by the difference between the maximum and minimum pressure (Pmax – Pmin). RVω showed significant negative correlations with the pulsed-wave Doppler-derived myocardial performance index (PWD-MPI) and the tissue Doppler imaging-derived MPI (TDI-MPI) (r = −0.52 and −0.51, respectively; both p < 0.0001). RVω also showed significant positive correlations with RV fractional area change (RVFAC) and RV ejection fraction (RVEF) (r = 0.41 and 0.39, respectively; both p < 0.0001), as well as a significant negative correlation with tricuspid E/e′ (r = −0.19, p = 0.0283). The clinical feasibility and utility of RVω for assessing global RV performance, incorporating both systolic and diastolic function, were demonstrated.
Ferromagnetism from non-magnetic ions: Ag-doped ZnO
Authors: Ali Nasir , A. R. Vijaya , Khan Zaheer Ahmed , Tarafder Kartick , Kumar Anuvesh , Wadhwa Manoj K. , Singh Budhi , Ghosh Subhasis
Published: December 2019

To develop suitable ferromagnetic oxides with Curie temperature (TC) at or above room temperature for spintronic applications, a great deal of research in doping ZnO with magnetic ions is being carried out over last decade. As the experimental results on magnetic ions doped ZnO are highly confused and controversial, we have investigated ferromagnetism in non-magnetic ion, Ag, doped ZnO. When Ag replaces Zn in ZnO, it adopts 4d9 configuration for Ag2+ which has single unpaired spin and suitable exchange interaction among these spins gives rise to ferromagnetism in ZnO with above room temperature TC. Experimentally, we have observed room temperature ferromagnetism (RTFM) in Ag-doped ZnO with Ag concentration varied from 0.03% to 10.0%. It is shown that zinc vacancy (VZn) enhances the ferromagnetic ordering (FMO) while oxygen vacancy (VO) retards the ferromagnetism in Ag-doped ZnO. Furthermore, the theoretical investigation revealed that VZn along with Ag2+ ions play a pivotal role for RTFM in Ag-doped ZnO. The Ag2+-Ag2+ interaction is ferromagnetic in the same Zn plane whereas anti-ferromagnetic in different Zn planes. The presence of VZn changes the anti-ferromagnetic to ferromagnetic state with a magnetic coupling energy of 37 meV. Finally, it has been established that the overlapping of bound magnetic polarons is responsible for RTFM in low doping concentration. However, anti-ferromagnetic coupling sets in at higher doping concentrations and hence weakens the FMO to a large extent.
Association between Serum Gamma-glutamyl transferase and Intracranial Arterial Calcification in Acute Ischemic Stroke Subjects
Authors: Yao Tao , Li Jing , Long Qi , Li Gang , Ding Yanbin , Cui Qin , Liu Zhichao
Published: December 2019

Intracranial artery calcification (IAC) is an important risk factor for cerebral infarction and a key biomarker for intracranial artery stenosis. Gamma-glutamyl transferase (GGT) has been independently associated with increased cardiovascular events and coronary calcification. Our study assessed whether GGT is an independent factor for IAC in acute ischemic stroke (AIS) patients. This cross-sectional study involved a total of 754 patients with AIS (mean age: 65 ± 13.2 years). All the patients had received brain computed tomography angiography (CTA) examination to evaluate IAC. Further, serum GGT levels and other biochemical parameters were analyzed. The average GGT level in patients who died was also significantly increased (37.0 ± 26.8 vs 29.0 ± 21.5 U/L, p = 0.012). Partial correlation analysis showed that serum GGT levels were associated with NIHSS score at admission after adjustment for age and gender was considered (r = 0.150, p = 0.001). Logistic regression analysis showed that serum GGT levels independently predicted all-cause mortality (OR = 1.036, 95% CI: 1.014–1.060, p = 0.002), NIHSS scores (β = 0.051, 95% CI: 0.020–0.082, p = 0.001) and IAC scores (β = 0.006, 95% CI: 0.003–0.014, p = 0.005) in male patients. Each SD (standard deviation) increase of serum GGT levels was also associated with risk of all-cause mortality (OR = 2.272, 95% CI: 1.364–3.787, P = 0.002). GGT levels in patients with severe IAC were significantly elevated (37.6 ± 33.6 vs 28.6 ± 19.2, p < 0.001). However, serum GGT levels could not independently predict the severity of IAC in AIS patients. Our study identified that serum GGT levels were significantly elevated in patients who died, and the GGT levels had a certain association with the risk of death and IAC in male patients.
Dimorphic metabolic and endocrine disorders in mice lacking the constitutive androstane receptor
Authors: Lukowicz Céline , Ellero-Simatos Sandrine , Régnier Marion , Oliviero Fabiana , Lasserre Frédéric , Polizzi Arnaud , Montagner Alexandra , Smati Sarra , Boudou Frédéric , Lenfant Françoise , Guzylack-Pirou Laurence , Menard Sandrine , Barretto Sharon , Fougerat Anne , Lippi Yannick , Naylies Claire , Bertrand-Michel Justine , Belgnaoui Afifa Ait , Theodorou Vassilia , Marchi Nicola , Gourdy Pierre , Gamet-Payrastre Laurence , Loiseau Nicolas , Guillou Hervé , Mselli-Lakhal Laïla
Published: December 2019

Metabolic diseases such as obesity, type II diabetes and hepatic steatosis are a public health concern in developed countries. The metabolic risk is gender‐dependent. The constitutive androstane receptor (CAR), which is at the crossroads between energy metabolism and endocrinology, has recently emerged as a promising therapeutic agent for the treatment of obesity and type 2 diabetes. In this study we sought to determine its role in the dimorphic regulation of energy homeostasis. We tracked male and female WT and CAR deficient (CAR−/−) mice for over a year. During aging, CAR−/− male mice developed hypercortisism, obesity, glucose intolerance, insulin insensitivity, dyslipidemia and hepatic steatosis. Remarkably, the latter modifications were absent, or minor, in female CAR−/− mice. When ovariectomized, CAR−/− female mice developed identical patterns of metabolic disorders as observed in male mice. These results highlight the importance of steroid hormones in the regulation of energy metabolism by CAR. They unveil a sexually dimorphic role of CAR in the maintenance of endocrine and metabolic homeostasis underscoring the importance of considering sex in treatment of metabolic diseases.
Combined use of a microbial restoration substrate and avirulent Ralstonia solanacearum for the control of tomato bacterial wilt
Authors: Zheng Xuefang , Zhu Yujing , Wang Jieping , Wang Ziran , Liu Bo
Published: December 2019

Tomato bacterial wilt (BW) caused by Ralstonia solanacearum seriously restricts tomato production and no effective control measures are available. A microbial restoration substrate (MRS) had been proved to be effective control of tomato BW in a greenhouse cultivation. In this study, MRS was combined with an avirulent Ralstonia solanacearum (aRS) strain to control the disease under an open field condition. In the two consecutive year (2017 and 2018) trials, the combined use of aRS and MRS resulted in better disease control compared with either aRS or MRS alone. Moreover, the combined treatment was more effective than expected and suggesting a synergistic control effect. Compared with control (CK, non-aRS or MRS), the application of aRS and MRS treatments alone or in combination could all promote plant growth, increase root activity and yield (e.g. the yield for the treatment of aRS + MRS increased by 463.64% in 2017). Soil nutrients, including soil organic carbon, total nitrogen, total phosphorus and total potassium contents were also significantly increased by the application of aRS and MRS treatments alone or in combination (P < 0.05). The application of MRS or in combination with aRS changed the soil from acidic to neutral, which is one of the key factors for controlling BW. The soil enzymatic activities were notably influenced by the combined use of aRS and MRS, which increased urease (87.37% in 2017 and 60.89% in 2018), catalase (93.67% in 2017 and 279.37% in 2018) and alkaline phosphatase activities (193.77% in 2017 and 455.73% in 2018). These results suggest that the combination of MRS and aRS could effectively control tomato BW and thus represents a promising new tool to control this disease.
Comorbid Medical Conditions as Predictors of Overall Survival in Glioblastoma Patients
Authors: Carr Matthew T. , Hochheimer Camille J. , Rock Andrew K. , Dincer Alper , Ravindra Lakshmi , Zhang Fan Lily , Opalak Charles F. , Poulos Nora , Sima Adam P. , Broaddus William C.
Published: December 2019

Glioblastoma (GBM) is an aggressive central nervous system tumor with a poor prognosis. This study was conducted to determine any comorbid medical conditions that are associated with survival in GBM. Data were collected from medical records of all patients who presented to VCU Medical Center with GBM between January 2005 and February 2015. Patients who underwent surgery/biopsy were considered for inclusion. Cox proportional hazards regression modeling was performed to assess the relationship between survival and sex, race, and comorbid medical conditions. 163 patients met inclusion criteria. Comorbidities associated with survival on individual-characteristic analysis included: history of asthma (Hazard Ratio [HR]: 2.63; 95% Confidence Interval [CI]: 1.24–5.58; p = 0.01), hypercholesterolemia (HR: 1.95; 95% CI: 1.09–3.50; p = 0.02), and incontinence (HR: 2.29; 95% CI: 0.95–5.57; p = 0.07). History of asthma (HR: 2.22; 95% CI: 1.02–4.83; p = 0.04) and hypercholesterolemia (HR: 1.99; 95% CI: 1.11–3.56; p = 0.02) were associated with shorter survival on multivariable analysis. Surgical patients with GBM who had a prior history of asthma or hypercholesterolemia had significantly higher relative risk for mortality on individual-characteristic and multivariable analyses.
Proline improves switchgrass growth and development by reduced lignin biosynthesis
Authors: Guan Cong , Cen Hui-Fang , Cui Xin , Tian Dan-Yang , Tadesse Dimiru , Zhang Yun-Wei
Published: December 2019

Transgenic switchgrass overexpressing Lolium perenne L. delta1-pyrroline 5-carboxylate synthase (LpP5CS) in group I (TG4 and TG6 line) and group II (TG1 and TG2 line) had significant P5CS and ProDH enzyme activities, with group I plants (TG4 and TG6) having higher P5CS and lower ProDH enzyme activity, while group II plants had higher ProDH and lower P5CS enzyme activity. We found group II transgenic plants showed stunted growth, and the changed proline content in overexpressing transgenic plants may influence the growth and development in switchgrass. RNA-seq analysis showed that KEGG enrichment included phenylpropanoid biosynthesis pathway among group I, group II and WT plants, and the expression levels of genes related to lignin biosynthesis were significantly up-regulated in group II. We also found that lignin content in group II transgenic plants was higher than that in group I and WT plants, suggesting that increased lignin content may suppress switchgrass growth and development. This study uncover that proline can appropriately reduce lignin biosynthesis to improve switchgrass growth and development. Therefore, appropriate reduction in lignin content and increase in biomass are important for bioenergy crop to lower processing costs for biomass fermentation-derived fuels.
Distinct pathways for zinc metabolism in the terrestrial slug Arion vulgaris
Authors: Dvorak Martin , Schnegg Raimund , Salvenmoser Willy , Palacios Òscar , Lindner Herbert , Zerbe Oliver , Hansel Armin , Leiminger Markus , Steiner Gerhard , Dallinger Reinhard , Lackner Reinhard
Published: December 2019

In most organisms, the concentration of free Zn2+ is controlled by metallothioneins (MTs). In contrast, no significant proportions of Zn2+ are bound to MTs in the slug, Arion vulgaris. Instead, this species possesses cytoplasmic low-molecular-weight Zn2+ (LMW Zn) binding compound that divert these metal ions into pathways uncoupled from MT metabolism. Zn2+ is accumulated in the midgut gland calcium cells of Arion vulgaris, where they associate with a low-molecular-weight ligand with an apparent molecular mass of ~ 2,000 Da. Mass spectrometry of the semi-purified LMW Zn binding compound combining an electrospray ion source with a differential mobility analyser coupled to a time-of-flight mass spectrometer revealed the presence of four Zn2+-containing ion signals, which arise from disintegration of one higher MW complex resulting in an ion-mobility diameter of 1.62 nm and a molecular mass of 837 Da. We expect that the novel Zn2+ ion storage pathway may be shared by many other gastropods, and particularly species that possess Cd-selective MT isoforms or variants with only very low affinity to Zn2+.
Identification of an optimal method for extracting RNA from human skin biopsy, using domestic pig as a model system
Authors: Reimann Ene , Abram Kristi , Kõks Sulev , Kingo Külli , Fazeli Alireza
Published: December 2019

To evaluate skin tissue gene expression patterns correctly, extracting sufficient quantities of good quality RNA is essential. However, RNA extraction from skin tissue is challenging, as the hyaluronic acid-collagen matrix is extremely difficult to homogenize. Although there are multiple ways to extract RNA from skin, there are no comparative studies that identify the most critical steps, e.g. sample collection, storage and homogenization. We analysed the various steps involved in RNA extraction (i.e. biopsy collection as dry biopsy or into nucleotide stabilizing reagents, different storage conditions, enzymatic digestion, stator-rotor and bead motion-based homogenizing combined with column-based RNA purification). We hypothesised that domestic pig skin is applicable as a model for human skin studies. Altogether twenty different workflows were tested on pig skin and the four most promising workflows were tested on human skin samples. The optimal strategy for extracting human skin RNA was to collect, store and homogenize the sample in RLT lysis buffer from the RNeasy Fibrous Tissue Kit combined with beta-mercaptoethanol. Both stator-rotor and bead motion-based homogenizing were found to result in high quality and quantity of extracted RNA. Our results confirmed that domestic pig skin can be successfully used as a model for human skin RNA studies.
Stepped vitrification technique for human ovarian tissue cryopreservation
Authors: Leonel Ellen Cristina Rivas , Corral Ariadna , Risco Ramon , Camboni Alessandra , Taboga Sebastião Roberto , Kilbride Peter , Vazquez Marina , Morris John , Dolmans Marie-Madeleine , Amorim Christiani A.
Published: December 2019

The advantage of stepped vitrification (SV) is avoiding ice crystal nucleation, while decreasing the toxic effects of high cryoprotectant concentrations. We aimed to test this method for human ovarian tissue cryopreservation. Ovarian cortex was taken from 7 fertile adult women. Samples were subjected to an SV protocol performed in an automatic freezer, which allowed sample transfer to ever higher concentrations of dimethyl sulfoxide (DMSO) as the temperature was reduced. Histological evaluation of the vitrified-warmed tissue showed large numbers of degenerated follicles after 24 hours of in vitro culture. We therefore evaluated DMSO perfusion rates by X-ray computed tomography, ice crystal formation by freeze-substitution, and cell toxicity by transmission electron microscopy, seeking possible reasons why follicles degenerated. Although cryoprotectant perfusion was considered normal and no ice crystals were formed in the tissue, ultrastructural analysis detected typical signs of DMSO toxicity, such as mitochondria degeneration, alterations in chromatin condensation, cell vacuolization and extracellular matrix swelling in both stromal and follicular cells. The findings indicated that the method failed to preserve follicles due to the high concentrations of DMSO used. However, adaptations can be made to avoid toxicity to follicles caused by elevated levels of cryoprotectants.
Chest Movement and Respiratory Volume both Contribute to Thoracic Bioimpedance during Loaded Breathing
Authors: Blanco-Almazán Dolores , Groenendaal Willemijn , Catthoor Francky , Jané Raimon
Published: December 2019

Bioimpedance has been widely studied as alternative to respiratory monitoring methods because of its linear relationship with respiratory volume during normal breathing. However, other body tissues and fluids contribute to the bioimpedance measurement. The objective of this study is to investigate the relevance of chest movement in thoracic bioimpedance contributions to evaluate the applicability of bioimpedance for respiratory monitoring. We measured airflow, bioimpedance at four electrode configurations and thoracic accelerometer data in 10 healthy subjects during inspiratory loading. This protocol permitted us to study the contributions during different levels of inspiratory muscle activity. We used chest movement and volume signals to characterize the bioimpedance signal using linear mixed-effect models and neural networks for each subject and level of muscle activity. The performance was evaluated using the Mean Average Percentage Errors for each respiratory cycle. The lowest errors corresponded to the combination of chest movement and volume for both linear models and neural networks. Particularly, neural networks presented lower errors (median below 4.29%). At high levels of muscle activity, the differences in model performance indicated an increased contribution of chest movement to the bioimpedance signal. Accordingly, chest movement contributed substantially to bioimpedance measurement and more notably at high muscle activity levels.
A systems biology approach reveals neuronal and muscle developmental defects after chronic exposure to ionising radiation in zebrafish
Authors: Murat El Houdigui Sophia , Adam-Guillermin Christelle , Loro Giovanna , Arcanjo Caroline , Frelon Sandrine , Floriani Magali , Dubourg Nicolas , Baudelet Emilie , Audebert Stéphane , Camoin Luc , Armant Olivier
Published: December 2019

Contamination of the environment after the Chernobyl and Fukushima Daiichi nuclear power plant (NPP) disasters led to the exposure of a large number of humans and wild animals to radioactive substances. However, the sub-lethal consequences induced by these absorbed radiological doses remain understudied and the long-term biological impacts largely unknown. We assessed the biological effects of chronic exposure to ionizing radiation (IR) on embryonic development by exposing zebrafish embryo from fertilization and up to 120 hours post-fertilization (hpf) at dose rates of 0.5 mGy/h, 5 mGy/h and 50 mGy/h, thereby encompassing the field of low dose rates defined at 6 mGy/h. Chronic exposure to IR altered larval behaviour in a light-dark locomotor test and affected cardiac activity at a dose rate as low as 0.5 mGy/h. The multi-omics analysis of transcriptome, proteome and transcription factor binding sites in the promoters of the deregulated genes, collectively points towards perturbations of neurogenesis, muscle development, and retinoic acid (RA) signaling after chronic exposure to IR. Whole-mount RNA in situ hybridization confirmed the impaired expression of the transcription factors her4.4 in the central nervous system and myogenin in the developing muscles of exposed embryos. At the organ level, the assessment of muscle histology by transmission electron microscopy (TEM) demonstrated myofibers disruption and altered neuromuscular junctions in exposed larvae at 5 mGy/h and 50 mGy/h. The integration of these multi-level data demonstrates that chronic exposure to low dose rates of IR has an impact on neuronal and muscle progenitor cells, that could lead to motility defects in free swimming larvae at 120 hpf. The mechanistic understanding of these effects allows us to propose a model where deregulation of RA signaling by chronic exposure to IR has pleiotropic effects on neurogenesis and muscle development.
Mn-TAT PTD-Ngb attenuates oxidative injury by an enhanced ROS scavenging ability and the regulation of redox signaling pathway
Authors: Zhang Cui , Hao Xuehui , Chang Jiaying , Geng Zhirong , Wang Zhilin
Published: December 2019

Neurological diseases have a close relationship to excessive reactive oxygen species (ROS). Neuroglobin (Ngb), an intrinsic protective factor, protected cells from hypoxic/ischemic injury. In the present, we reported a novel neuroprotective manganese porphyrin reconstituted metal protein, Mn-TAT PTD-Ngb, consisting of a HIV Tat protein transduction domain sequence (TAT PTD) attached to the N-terminal of apo-Ngb. Mn-TAT PTD-Ngb had a stronger ROS scavenging ability than that of TAT PTD-Ngb, and reduced intracellular ROS production and restored the function of the mitochondria and inhibited the mitochondria-dependent apoptosis. Besides, Mn-TAT PTD-Ngb activated the phosphoinositide-3 kinase (PI3K)/Akt signaling pathway, which up-regulated the expression of nuclear factor E2-related factor 2 (Nrf2), Heme oxygenase-1 (HO-1), superoxide dismutase (SOD), catalase (CAT). The results showed that the redox chemistry of Mn-TAT PTD-Ngb and redox regulation of multiple signaling pathways attenuated the oxidative injury.
The Relation between Alpha/Beta Oscillations and the Encoding of Sentence induced Contextual Information
Authors: Terporten René , Schoffelen Jan-Mathijs , Dai Bohan , Hagoort Peter , Kösem Anne
Published: December 2019

Pre-stimulus alpha (8–12 Hz) and beta (16–20 Hz) oscillations have been frequently linked to the prediction of upcoming sensory input. Do these frequency bands serve as a neural marker of linguistic prediction as well? We hypothesized that if pre-stimulus alpha and beta oscillations index language predictions, their power should monotonically relate to the degree of predictability of incoming words based on past context. We expected that the more predictable the last word of a sentence, the stronger the alpha and beta power modulation. To test this, we measured neural responses with magnetoencephalography of healthy individuals during exposure to a set of linguistically matched sentences featuring three levels of sentence context constraint (high, medium and low constraint). We observed fluctuations in alpha and beta power before last word onset, and modulations in M400 amplitude after last word onset. The M400 amplitude was monotonically related to the degree of context constraint, with a high constraining context resulting in the strongest amplitude decrease. In contrast, pre-stimulus alpha and beta power decreased more strongly for intermediate constraints, followed by high and low constraints. Therefore, unlike the M400, pre-stimulus alpha and beta dynamics were not indexing the degree of word predictability from sentence context.
The effect of ligand amount, affinity and internalization on PSMA-targeted imaging and therapy: A simulation study using a PBPK model
Authors: Begum Nusrat J. , Glatting Gerhard , Wester Hans-Jürgen , Eiber Matthias , Beer Ambros J. , Kletting Peter
Published: December 2019

The aim of this work was to investigate the effect of ligand amount, affinity and internalization of prostate-specific membrane antigen (PSMA)-specific ligands on the activity concentrations for PET/CT imaging and on the absorbed doses for therapy. A physiologically-based pharmacokinetic (PBPK) model for PSMA-specific ligands was implemented. Thirteen virtual patients with metastatic castration-resistant prostate cancer were analysed. Simulations were performed for different combinations of association rates kon (0.1–0.01 L/nmol/min), dissociation rates koff (0.1–0.0001 min−1), internalization rates λint (0.01–0.0001 min−1) and ligand amounts (1–1000 nmol). For imaging the activity was normalized to volume and injected activity (68Ga-PSMA at 1 h). For therapy the absorbed dose was calculated for 7.3 ± 0.3 GBq 177Lu-PSMA. The effect of the investigated parameters on therapy were larger compared to imaging. For imaging, the combination of properties leading to the highest tumour uptake was kon = 0.1 L/nmol/min, koff = 0.01 min−1 for typical ligand amounts (1–10 nmol). For therapy, the higher the internalization rate, the larger was the required ligand amount for optimal tumour-to-kidney ratios. The higher the affinity, the more important was the choice of the optimal ligand amount. PBPK modelling provides insight into the pharmacokinetics of PSMA-specific ligands. Further in silico and in vivo studies are required to verify the influence of the analysed parameters.
An improved simple method for the identification of Mycobacteria by MALDI-TOF MS (Matrix-Assisted Laser Desorption- Ionization mass spectrometry)
Authors: Alcolea-Medina Adela , Fernandez M. T. Cabezas , Montiel N. , García M. P. Luzón , Sevilla C. Delamo , North Nathan , Lirola M. J. Martínez , Wilks Mark
Published: December 2019

The aim of this study was to establish a simple method for the rapid identification of Mycobacteria species by MALDI-TOF (Matrix-Assisted Laser Desorption/Ionization-Time of Flight Mass spectrometry) using the Bruker MALDI-TOF Biotyper system (Bruker Daltonik, Bremen, Germany). A multicentre, prospective, and single blind study was performed in three European Hospitals, two Spanish and one UK hospital from May to August 2018. The BD BACTEC MGIT (Becton Dickinson, Berks, UK) liquid culture system was used in all three centres for the growth of Mycobacteria. When signal positive, tubes were removed from the analyser and in addition to standard laboratory procedures were subcultured on blood agar plates for MALDI-TOF analysis. Plates were incubated aerobically for 1 to 7 days at 37 °C and inspected every day. Once any growth was visible, it was transferred to the steel target plate, overlaid with 1 μl of neat formic acid and 1 μl HCCA matrix (alpha hydroxyl 4 cinnamic acid), and analysed in a Bruker Biotyper MALDI-TOF. Results given by MALDI-TOF were compared with the reference methods used for identification in the different centres. At two Spanish hospitals, identification by MALDI-TOF was only attempted on presumptive non-tuberculosis mycobacteria (NTM) and the results were initially compared with the results obtained by a commercial reverse hybridisation assay, GenoType CM/AS (Hain Lifescience, Tübingen, Germany). At the UK Hospital, identification of any presumptive mycobacteria was attempted and compared with the results obtained by whole genome sequencing (WGS). Overall in 142/167 (85%) of cases the identifications obtained were concordant; all Mycobacterium tuberculosis (MTB) isolates 43/43 (100%), 57/76 (75%) of the rapid growing nontuberculous mycobacteria (NTM), and 42/48 (85%) slow growing NTM tested were identified correctly. We report a new, easy, cheap and quick method for isolation and identification of Mycobacterium spp. without the need for additional steps or equipment and this method is in routine used in all three centres.
Experimental evidence of mechanical lumpy skin disease virus transmission by Stomoxys calcitrans biting flies and Haematopota spp. horseflies
Authors: Sohier C. , Haegeman A. , Mostin L. , De Leeuw I. , Campe W. Van , De Vleeschauwer A. , Tuppurainen E. S. M. , van den Berg T. , De Regge N. , De Clercq K.
Published: December 2019

Lumpy skin disease (LSD) is a devastating disease of cattle characterized by fever, nodules on the skin, lymphadenopathy and milk drop. Several haematophagous arthropod species like dipterans and ticks are suspected to play a role in the transmission of LSDV. Few conclusive data are however available on the importance of biting flies and horseflies as potential vectors in LSDV transmission. Therefore an in vivo transmission study was carried out to investigate possible LSDV transmission by Stomoxys calcitrans biting flies and Haematopota spp. horseflies from experimentally infected viraemic donor bulls to acceptor bulls. LSDV transmission by Stomoxys calcitrans was evidenced in 3 independent experiments, LSDV transmission by Haematopota spp. was shown in one experiment. Evidence of LSD was supported by induction of nodules and virus detection in the blood of acceptor animals. Our results are supportive for a mechanical transmission of the virus by these vectors.
Normal GCAPs partly compensate for altered cGMP signaling in retinal dystrophies associated with mutations in GUCA1A
Authors: Dell’Orco Daniele , Dal Cortivo Giuditta
Published: December 2019

Missense mutations in the GUCA1A gene encoding guanylate cyclase-activating protein 1 (GCAP1) are associated with autosomal dominant cone/cone-rod (CORD) dystrophies. The nature of the inheritance pattern implies that a pool of normal GCAP proteins is present in photoreceptors together with the mutated variant. To assess whether human GCAP1 and GCAP2 may similarly regulate the activity of the retinal membrane guanylate cyclase GC-1 (GC-E) in the presence of the recently discovered E111V-GCAP1 CORD-variant, we combined biochemical and in silico assays. Surprisingly, human GCAP2 does not activate GC1 over the physiological range of Ca2+ whereas wild-type GCAP1 significantly attenuates the dysregulation of GC1 induced by E111V-GCAP1. Simulation of the phototransduction cascade in a well-characterized murine system, where GCAP2 is able to activate the GC1, suggests that both GCAPs can act in a synergic manner to mitigate the effects of the CORD-mutation. We propose the existence of a species-dependent compensatory mechanism. In murine photoreceptors, slight increases of wild-type GCAPs levels may significantly attenuate the increase in intracellular Ca2+ and cGMP induced by E111V-GCAP1 in heterozygous conditions. In humans, however, the excess of wild-type GCAP1 may only partly attenuate the mutant-induced dysregulation of cGMP signaling due to the lack of GC1-regulation by GCAP2.
Zebrafish structural development in Mueller-matrix scanning microscopy
Authors: Le Gratiet Aymeric , d’Amora Marta , Duocastella Marti , Marongiu Riccardo , Bendandi Artemi , Giordani Silvia , Bianchini Paolo , Diaspro Alberto
Published: December 2019

Zebrafish are powerful animal models for understanding biological processes and the molecular mechanisms involved in different human diseases. Advanced optical techniques based on fluorescence microscopy have become the main imaging method to characterize the development of these organisms at the microscopic level. However, the need for fluorescence probes and the consequent high light doses required to excite fluorophores can affect the biological process under observation including modification of metabolic function or phototoxicity. Here, without using any labels, we propose an implementation of a Mueller-matrix polarimeter into a commercial optical scanning microscope to characterize the polarimetric transformation of zebrafish preserved at different embryonic developmental stages. By combining the full polarimetric measurements with statistical analysis of the Lu and Chipman mathematical decomposition, we demonstrate that it is possible to quantify the structural changes of the biological organization of fixed zebrafish embryos and larvae at the cellular scale. This convenient implementation, with low light intensity requirement and cheap price, coupled with the quantitative nature of Mueller-matrix formalism, can pave the way for a better understanding of developmental biology, in which label-free techniques become a standard tool to study organisms.
High degree of polyclonality hinders somatic mutation calling in lung brush samples of COPD cases and controls
Authors: Thun Gian-Andri , Derdak Sophia , Castro-Giner Francesc , Apunte-Ramos Katherine , Águeda Lidia , Wjst Matthias , Boland Anne , Deleuze Jean-François , Kolsum Umme , Heiss-Neumann Marion S. , Nowinski Adam , Gorecka Dorota , Hohlfeld Jens M. , Welte Tobias , Brightling Christopher E. , Parr David G. , Prasse Antje , Müller-Quernheim Joachim , Greulich Timm , Stendardo Mariarita , Boschetto Piera , Barta Imre , Döme Balázs , Gut Marta , Singh Dave , Ziegler-Heitbrock Loems , Gut Ivo G.
Published: December 2019

Chronic obstructive pulmonary disease (COPD) is induced by cigarette smoking and characterized by inflammation of airway tissue. Since smokers with COPD have a higher risk of developing lung cancer than those without, we hypothesized that they carry more mutations in affected tissue. We called somatic mutations in airway brush samples from medium-coverage whole genome sequencing data from healthy never and ex-smokers (n = 8), as well as from ex-smokers with variable degrees of COPD (n = 4). Owing to the limited concordance of resulting calls between the applied tools we built a consensus, a strategy that was validated with high accuracy for cancer data. However, consensus calls showed little promise of representing true positives due to low mappability of corresponding sequence reads and high overlap with positions harbouring known genetic polymorphisms. A targeted re-sequencing approach suggested that only few mutations would survive stringent verification testing and that our data did not allow the inference of any difference in the mutational load of bronchial brush samples between former smoking COPD cases and controls. High polyclonality in airway brush samples renders medium-depth sequencing insufficient to provide the resolution to detect somatic mutations. Deep sequencing data of airway biopsies are needed to tackle the question.
Eye lens radiation exposure in paediatric interventional treatment of retinoblastoma
Authors: Obesso A. , Alejo L. , Huerga C. , Sánchez-Muñoz F. , Corredoira E. , Fernández-Prieto A. , Frutos R. , Marín B. , Garzón G. , Peralta J. , Ubeda C. , Guibelalde E.
Published: December 2019

Retinoblastoma represents 3% of cancers in children under fifteen years of age. The standard paediatric treatment for saving the affected eye is supraselective intra-arterial chemotherapy performed in interventional rooms. In order to address the radiation toxicity due to the angiography, the aim of this study was to determine the typical dose value corresponding to the procedure, estimate the paediatric patients’ eye lens dose and study the relationship between dose indicators and dose to the lens. An automatic dose management software was installed in two interventional rooms to obtain the distribution of the dose indicators kerma-area product and reference-point air kerma, getting a typical value 16 Gy·cm2 and 130 mGy, respectively (n = 35). The eye lens dose estimates were obtained with photoluminescent dosimeters placed on the patient’s eyelids. In the left eye, the entrance surface air kerma was 44.23 ± 2.66 mGy, and 12.72 ± 0.89 mGy in the right eye (n = 10). There was a positive correlation between dose to the lens per procedure and dose indicators, with R2 > 0.65 for both eyes. Based on this information, the threshold for the onset of radiation-induce cataracts (500 mGy) will be exceeded if the treatment is performed for more than 8 sessions.
Relation between platelet coagulant and vascular function, sex-specific analysis in adult survivors of childhood cancer compared to a population-based sample
Authors: Panova-Noeva Marina , Wagner Bianca , Nagler Markus , Arnold Natalie , Prochaska Jürgen H. , Eckerle Susan , Spronk Henri M. , Merzenich Hiltrud , Wingerter Arthur , Schneider Astrid , Danckwardt Sven , ten Cate Hugo , Faber Jörg , Wild Philipp S.
Published: December 2019

Female sex is a risk factor for long-term adverse outcome in cancer survivors, however very little is known for the underlying pathophysiological mechanisms rendering the increased risk. This study investigated sex-specifically the relation between thrombin generation (TG) with and without presence of platelets and vascular function in 200 adult survivors of a childhood cancer compared to 335 population-based control individuals. TG lag time, peak height and endogenous thrombin potential (ETP) measured in presence and absence of platelets were correlated to reflection index (RI) and stiffness index (SI). A sex-specific correlation analysis showed a negative relation in female survivors for platelet-dependent peak height and/or ETP and RI only. An age adjusted linear regression model confirmed the negative association between RI and platelet-dependent ETP (beta estimate: −6.85, 95% confidence interval: −12.19,−1.51) in females. Adjustment for cardiovascular risk factors resulted in loss of the association, whereby arterial hypertension and obesity showed the largest effects on the observed association. No other relevant associations were found in male and female cancer survivors and all population-based controls. This study demonstrates a link between platelet coagulant and vascular function of resistance vessels, found in female cancer survivors, potentially mediated by the presence of arterial hypertension and obesity.
An acute session of motor imagery training induces use-dependent plasticity
Authors: Ruffino Célia , Gaveau Jérémie , Papaxanthis Charalambos , Lebon Florent
Published: December 2019

Motor imagery, defined as the mental representation of an action without movement-related sensory inputs, is a well-known intervention to improve motor performance. In the current study, we tested whether use-dependent plasticity, a mechanism underlying motor learning, could be induced by an acute session of motor imagery. By means of transcranial magnetic stimulation (TMS) over the left primary motor cortex, we evoked isolated thumb movements in the right hand and assessed corticospinal excitability in the flexor and extensor pollicis brevis muscles. We measured the mean TMS-induced movement direction before and after an acute session of motor imagery practice. In a first experiment, participants of the imagery group were instructed to repeatedly imagine their thumb moving in a direction deviated by 90° from the pre-test movement. This group, but not the control group, deviated the post-training TMS-induced movements toward the training target direction (+44° ± 62° and −1° ± 23°, respectively). Interestingly, the deviation magnitude was driven by the corticospinal excitability increase in the agonist muscle. In a second experiment, we found that post-training TMS-induced movements were proportionally deviated toward the trained direction and returned to baseline 30 minutes after the motor imagery training. These findings suggest that motor imagery induces use-dependent plasticity and, this neural process is accompanied by corticospinal excitability increase in the agonist muscle.