bluuebunny/crossref_metadata_2025 huggingface dataset:
Landscape change patterns at three stages of the construction and operation of the TGP
Authors: Li Ruikang , Li Yangbing , Li Bo , Fu Dianji
Published: April 2021

Analyses of landscape change patterns that are based on elevation and slope can not only provide reasonable interpretations of landscape patterns but can also help to reveal evolutionary laws. However, landscape change patterns and their model in different landforms of the typical watershed in the Three Gorges Reservoir Area (TGRA) has not been quantified and assessed effectively. As a complex geographical unit, the ecological environment in the middle reach of the Yangtze River has experienced great changes due to the construction of the Three Gorges Project (TGP) and its associated human activities. Here, based mainly on a digital elevation model (DEM) and remotely sensed images from 1986, 2000, 2010, and 2017 and by using GIS technology, speeds/ trends of landscape change, the index of landscape type change intensity, landscape pattern indices, and landscape ecological security index, the spatial and temporal evolution characteristics of different elevations, slopes, and buffer landscape types were analyzed in typical watersheds, as well as an evolutionary model of the landscape pattern. The results indicated that (1) the landscape types along with the land classification and buffer zone that were influenced by the TGR construction have undergone a phased change, with the period 2000–2010 being the most dramatic period of landscape evolution during the impoundment period; (2) landscape type shifts from human-dominated farmland to nature-driven forestland and shrub-land as elevations, slopes and buffer distances increased. The landscape has shifted from diversity to relative homogeneity; (3) land types and buffer zones played essential roles in the landscape pattern index, which is reflected in the differences in landscape type indices for spatial extension and temporal characteristics. The results of this paper illustrate the spatial–temporal characteristics of various landscape types at three distinct stages in the construction of the TGR. These findings indicate that the landscape ecological security of the watershed is improving year by year. The follow-up development of the TGRA needs to consider the landscape change patterns of different landforms.
Alantolactone inhibits cervical cancer progression by downregulating BMI1
Authors: Sun Xiaodong , Xu Hongxia , Dai Tianyu , Xie Lixia , Zhao Qiang , Hao Xincai , Sun Yan , Wang Xuanbin , Jiang Nan , Sang Ming
Published: April 2021

Cervical cancer is the second most common cancer in women. Despite advances in cervical cancer therapy, tumor recurrence and metastasis remain the leading causes of mortality. High expression of BMI1 is significantly associated with poor tumor differentiation, high clinical grade, and poor prognosis of cervical cancer, and is an independent prognostic factor in cervical carcinoma. Alantolactone (AL), a sesquiterpene lactone, exhibits potent anti-inflammatory and anticancer activities. In this paper, we investigated the mechanism of AL in reducing the proliferation, migration, and invasion of HeLa and SiHa cervical cancer cells as well as its promotion of mitochondrial damage and autophagy. BMI1 silencing decreased epithelial-mesenchymal transformation-associated proteins and increased autophagy-associated proteins in HeLa cells. These effects were reversed by overexpression of BMI1 in HeLa cells. Thus, BMI1 expression is positively correlated with invasion and negatively correlated with autophagy in HeLa cells. Importantly, AL decreased the weight, volume, and BMI1 expression in HeLa xenograft tumors. Furthermore, the structure of BMI1 and target interaction of AL were virtually screened using the molecular docking program Autodock Vina; AL decreased the expression of N-cadherin, vimentin, and P62 and increased the expression of LC3B and Beclin-1 in xenograft tumors. Finally, expression of BMI1 increased the phosphorylation of STAT3, which is important for cell proliferation, survival, migration, and invasion. Therefore, we suggest that AL plays a pivotal role in inhibiting BMI1 in the tumorigenesis of cervical cancer and is a potential therapeutic agent for cervical cancer.
Synchronized resistive-pulse analysis with flow visualization for single micro- and nanoscale objects driven by optical vortex in double orifice
Authors: Nakajima Kichitaro , Nakatsuka Ryoji , Tsuji Tetsuro , Doi Kentaro , Kawano Satoyuki
Published: April 2021

Resistive-pulse analysis is a powerful tool for identifying micro- and nanoscale objects. For low-concentration specimens, the pulse responses are rare, and it is difficult to obtain a sufficient number of electrical waveforms to clearly characterize the targets and reduce noise. In this study, we conducted a periodic resistive-pulse analysis using an optical vortex and a double orifice, which repetitively senses a single micro- or nanoscale target particle with a diameter ranging from 700 nm to 2 $$\mu$$
μ
m. The periodic motion results in the accumulation of a sufficient number of waveforms within a short period. Acquired pulses show periodic ionic-current drops associated with the translocation events through each orifice. Furthermore, a transparent fluidic device allows us to synchronously average the waveforms by the microscopic observation of the translocation events and improve the signal-to-noise ratio. By this method, we succeed in distinguishing single particle diameters. Additionally, the results of measured signals and the simultaneous high-speed observations are used to quantitatively and systematically discuss the effect of the complex fluid flow in the orifices on the amplitude of the resistive pulse. The synchronized resistive-pulse analysis by the optical vortex with the flow visualization improves the pulse-acquisition rate for a single specific particle and accuracy of the analysis, refining the micro- and nanoscale object identification.
Analyzing the impact of induced magnetic flux and Fourier’s and Fick’s theories on the Carreau-Yasuda nanofluid flow
Authors: Bashir Seemab , Ramzan Muhammad , Chung Jae Dong , Chu Yu-Ming , Kadry Seifedine
Published: April 2021

The current study analyzes the effects of modified Fourier and Fick's theories on the Carreau-Yasuda nanofluid flow over a stretched surface accompanying activation energy with binary chemical reaction. Mechanism of heat transfer is observed in the occurrence of heat source/sink and Newtonian heating. The induced magnetic field is incorporated to boost the electric conductivity of nanofluid. The formulation of the model consists of nonlinear coupled partial differential equations that are transmuted into coupled ordinary differential equations with high nonlinearity by applying boundary layer approximation. The numerical solution of this coupled system is carried out by implementing the MATLAB solver bvp4c package. Also, to verify the accuracy of the numerical scheme grid-free analysis for the Nusselt number is presented. The influence of different parameters, for example, reciprocal magnetic Prandtl number, stretching ratio parameter, Brownian motion, thermophoresis, and Schmidt number on the physical quantities like velocity, temperature distribution, and concentration distribution are addressed with graphs. The Skin friction coefficient and local Nusselt number for different parameters are estimated through Tables. The analysis shows that the concentration of nanoparticles increases on increasing the chemical reaction with activation energy and also Brownian motion efficiency and thermophoresis parameter increases the nanoparticle concentration. Opposite behavior of velocity profile and the Skin friction coefficient is observed for increasing the stretching ratio parameter. In order to validate the present results, a comparison with previously published results is presented. Also, Factors of thermal and solutal relaxation time effectively contribute to optimizing the process of stretchable surface chilling, which is important in many industrial applications.
Using supraglottic airways by paramedics for airway management in analogue microgravity increases speed and success of ventilation
Authors: Hinkelbein Jochen , Ahlbäck Anton , Antwerber Christine , Dauth Lisa , DuCanto James , Fleischhammer Elisabeth , Glatz Carlos , Kerkhoff Steffen , Mathes Alexander , Russomano Thais , Schmitz Jan , Starck Clement , Thierry Seamus , Warnecke Tobias
Published: April 2021

In the next few years, the number of long-term space missions will significantly increase. Providing safe concepts for emergencies including airway management will be a highly challenging task. The aim of the present trial is to compare different airway management devices in simulated microgravity using a free-floating underwater scenario. Five different devices for airway management [laryngeal mask (LM), laryngeal tube (LT), I-GEL, direct laryngoscopy (DL), and video laryngoscopy (VL)] were compared by n = 20 paramedics holding a diving certificate in a randomized cross-over setting both under free-floating conditions in a submerged setting (pool, microgravity) and on ground (normogravity). The primary endpoint was the successful placement of the airway device. The secondary endpoints were the number of attempts and the time to ventilation. A total of 20 paramedics (3 female, 17 male) participated in this study. Success rate was highest for LM and LT and was 100% both during simulated microgravity and normogravity followed by the I-GEL (90% during microgravity and 95% during normogravity). However, the success rate was less for both DL (60% vs. 95%) and VL (20% vs. 60%). Fastest ventilation was performed with the LT both in normogravity (13.7 ± 5.3 s; n = 20) and microgravity (19.5 ± 6.1 s; n = 20). For the comparison of normogravity and microgravity, time to ventilation was shorter for all devices on the ground (normogravity) as compared underwater (microgravity). In the present study, airway management with supraglottic airways and laryngoscopy was shown to be feasible. Concerning the success rate and time to ventilation, the optimum were supraglottic airways (LT, LM, I-GEL) as their placement was faster and associated with a higher success rate. For future space missions, the use of supraglottic airways for airway management seems to be more promising as compared to tracheal intubation by DL or VL.
Fleeing lockdown and its impact on the size of epidemic outbreaks in the source and target regions – a COVID-19 lesson
Authors: Barbarossa Maria Vittoria , Bogya Norbert , Dénes Attila , Röst Gergely , Varma Hridya Vinod , Vizi Zsolt
Published: April 2021

The COVID-19 pandemic forced authorities worldwide to implement moderate to severe restrictions in order to slow down or suppress the spread of the disease. It has been observed in several countries that a significant number of people fled a city or a region just before strict lockdown measures were implemented. This behavior carries the risk of seeding a large number of infections all at once in regions with otherwise small number of cases. In this work, we investigate the effect of fleeing on the size of an epidemic outbreak in the region under lockdown, and also in the region of destination. We propose a mathematical model that is suitable to describe the spread of an infectious disease over multiple geographic regions. Our approach is flexible to characterize the transmission of different viruses. As an example, we consider the COVID-19 outbreak in Italy. Projection of different scenarios shows that (i) timely and stricter intervention could have significantly lowered the number of cumulative cases in Italy, and (ii) fleeing at the time of lockdown possibly played a minor role in the spread of the disease in the country.
Distribution, associations and role in the biological carbon pump of Pyrosoma atlanticum (Tunicata, Thaliacea) off Cabo Verde, NE Atlantic
Authors: Stenvers Vanessa I. , Hauss Helena , Osborn Karen J. , Neitzel Philipp , Merten Véronique , Scheer Stella , Robison Bruce H. , Freitas Rui , Hoving Henk Jan T.
Published: April 2021

Gelatinous zooplankton are increasingly acknowledged to contribute significantly to the carbon cycle worldwide, yet many taxa within this diverse group remain poorly studied. Here, we investigate the pelagic tunicatePyrosoma atlanticumin the waters surrounding the Cabo Verde Archipelago. By using a combination of pelagic and benthic in situ observations, sampling, and molecular genetic analyses (barcoding, eDNA), we reveal that:P. atlanticumabundance is most likely driven by local island-induced productivity, that it substantially contributes to the organic carbon export flux and is part of a diverse range of biological interactions. Downward migrating pyrosomes actively transported an estimated 13% of their fecal pellets below the mixed layer, equaling a carbon flux of 1.96–64.55 mg C m−2 day−1. We show that analysis of eDNA can detect pyrosome material beyond their migration range, suggesting that pyrosomes have ecological impacts below the upper water column. MoribundP. atlanticumcolonies contributed an average of 15.09 ± 17.89 (s.d.) mg C m−2to the carbon flux reaching the island benthic slopes. Our pelagic in situ observations further show thatP. atlanticumformed an abundant substrate in the water column (reaching up to 0.28 m2substrate area per m2), with animals using pyrosomes for settlement, as a shelter and/or a food source. In total, twelve taxa from four phyla were observed to interact with pyrosomes in the midwater and on the benthos.
TRPM2 ion channels steer neutrophils towards a source of hydrogen peroxide
Authors: Morad Hassan , Luqman Suaib , Tan Chun-Hsiang , Swann Victoria , McNaughton Peter A.
Published: April 2021

Neutrophils must navigate accurately towards pathogens in order to destroy invaders and thus defend our bodies against infection. Here we show that hydrogen peroxide, a potent neutrophil chemoattractant, guides chemotaxis by activating calcium-permeable TRPM2 ion channels and generating an intracellular leading-edge calcium “pulse”. The thermal sensitivity of TRPM2 activation means that chemotaxis towards hydrogen peroxide is strongly promoted by small temperature elevations, suggesting that an important function of fever may be to enhance neutrophil chemotaxis by facilitating calcium influx through TRPM2. Chemotaxis towards conventional chemoattractants such as LPS, CXCL2 and C5a does not depend on TRPM2 but is driven in a similar way by leading-edge calcium pulses. Other proposed initiators of neutrophil movement, such as PI3K, Rac and lyn, influence chemotaxis by modulating the amplitude of calcium pulses. We propose that intracellular leading-edge calcium pulses are universal drivers of the motile machinery involved in neutrophil chemotaxis.
Machine learning outperforms thermodynamics in measuring how well a many-body system learns a drive
Authors: Zhong Weishun , Gold Jacob M. , Marzen Sarah , England Jeremy L. , Yunger Halpern Nicole
Published: April 2021

Diverse many-body systems, from soap bubbles to suspensions to polymers, learn and remember patterns in the drives that push them far from equilibrium. This learning may be leveraged for computation, memory, and engineering. Until now, many-body learning has been detected with thermodynamic properties, such as work absorption and strain. We progress beyond these macroscopic properties first defined for equilibrium contexts: We quantify statistical mechanical learning using representation learning, a machine-learning model in which information squeezes through a bottleneck. By calculating properties of the bottleneck, we measure four facets of many-body systems’ learning: classification ability, memory capacity, discrimination ability, and novelty detection. Numerical simulations of a classical spin glass illustrate our technique. This toolkit exposes self-organization that eludes detection by thermodynamic measures: Our toolkit more reliably and more precisely detects and quantifies learning by matter while providing a unifying framework for many-body learning.
Long-read-sequenced reference genomes of the seven major lineages of enterotoxigenic Escherichia coli (ETEC) circulating in modern time
Authors: von Mentzer Astrid , Blackwell Grace A. , Pickard Derek , Boinett Christine J. , Joffré Enrique , Page Andrew J. , Svennerholm Ann-Mari , Dougan Gordon , Sjöling Åsa
Published: April 2021

Enterotoxigenic Escherichia coli (ETEC) is an enteric pathogen responsible for the majority of diarrheal cases worldwide. ETEC infections are estimated to cause 80,000 deaths annually, with the highest rates of burden, ca 75 million cases per year, amongst children under 5 years of age in resource-poor countries. It is also the leading cause of diarrhoea in travellers. Previous large-scale sequencing studies have found seven major ETEC lineages currently in circulation worldwide. We used PacBio long-read sequencing combined with Illumina sequencing to create high-quality complete reference genomes for each of the major lineages with manually curated chromosomes and plasmids. We confirm that the major ETEC lineages all harbour conserved plasmids that have been associated with their respective background genomes for decades, suggesting that the plasmids and chromosomes of ETEC are both crucial for ETEC virulence and success as pathogens. The in-depth analysis of gene content, synteny and correct annotations of plasmids will elucidate other plasmids with and without virulence factors in related bacterial species. These reference genomes allow for fast and accurate comparison between different ETEC strains, and these data will form the foundation of ETEC genomics research for years to come.
Geometrically defined environments direct cell division rate and subcellular YAP localization in single mouse embryonic stem cells
Authors: Bertels Sarah , Jaggy Mona , Richter Benjamin , Keppler Stephan , Weber Kerstin , Genthner Elisa , Fischer Andrea C. , Thiel Michael , Wegener Martin , Greiner Alexandra M. , Autenrieth Tatjana J. , Bastmeyer Martin
Published: April 2021

Mechanotransduction via yes-associated protein (YAP) is a central mechanism for decision-making in mouse embryonic stem cells (mESCs). Nuclear localization of YAP is tightly connected to pluripotency and increases the cell division rate (CDR). How the geometry of the extracellular environment influences mechanotransduction, thereby YAP localization, and decision-making of single isolated mESCs is largely unknown. To investigate this relation, we produced well-defined 2D and 2.5D microenvironments and monitored CDR and subcellular YAP localization in single mESCs hence excluding cell–cell interactions. By systematically varying size and shape of the 2D and 2.5D substrates we observed that the geometry of the growth environment affects the CDR. Whereas CDR increases with increasing adhesive area in 2D, CDR is highest in small 2.5D micro-wells. Here, mESCs attach to all four walls and exhibit a cross-shaped cell and nuclear morphology. This observation indicates that changes in cell shape are linked to a high CDR. Inhibition of actomyosin activity abrogate these effects. Correspondingly, nuclear YAP localization decreases in inhibitor treated cells, suggesting a relation between cell shape, intracellular forces, and cell division rate. The simplicity of our system guarantees high standardization and reproducibility for monitoring stem cell reactions and allows addressing a variety of fundamental biological questions on a single cell level.
Multi-seasonal systematic camera-trapping reveals fluctuating densities and high turnover rates of Carpathian lynx on the western edge of its native range
Authors: Duľa Martin , Bojda Michal , Chabanne Delphine B. H. , Drengubiak Peter , Hrdý Ľuboslav , Krojerová-Prokešová Jarmila , Kubala Jakub , Labuda Jiří , Marčáková Leona , Oliveira Teresa , Smolko Peter , Váňa Martin , Kutal Miroslav
Published: April 2021

Camera-trapping and capture-recapture models are the most widely used tools for estimating densities of wild felids that have unique coat patterns, such as Eurasian lynx. However, studies dealing with this species are predominantly on a short-term basis and our knowledge of temporal trends and population persistence is still scarce. By using systematic camera-trapping and spatial capture-recapture models, we estimated lynx densities and evaluated density fluctuations, apparent survival, transition rate and individual's turnover during five consecutive seasons at three different sites situated in the Czech–Slovak–Polish borderland at the periphery of the Western Carpathians. Our density estimates vary between 0.26 and 1.85 lynx/100 km2 suitable habitat and represent the lowest and the highest lynx densities reported from the Carpathians. We recorded 1.5–4.1-fold changes in asynchronous fluctuated densities among all study sites and seasons. Furthermore, we detected high individual’s turnover (on average 46.3 ± 8.06% in all independent lynx and 37.6 ± 4.22% in adults) as well as low persistence of adults (only 3 out of 29 individuals detected in all seasons). The overall apparent survival rate was 0.63 ± 0.055 and overall transition rate between sites was 0.03 ± 0.019. Transition rate of males was significantly higher than in females, suggesting male-biased dispersal and female philopatry. Fluctuating densities and high turnover rates, in combination with documented lynx mortality, indicate that the population in our region faces several human-induced mortalities, such as poaching or lynx-vehicle collisions. These factors might restrict population growth and limit the dispersion of lynx to other subsequent areas, thus undermining the favourable conservation status of the Carpathian population. Moreover, our study demonstrates that long-term camera-trapping surveys are needed for evaluation of population trends and for reliable estimates of demographic parameters of wild territorial felids, and can be further used for establishing successful management and conservation measures.
Orexinergic descending inhibitory pathway mediates linalool odor-induced analgesia in mice
Authors: Higa Yurina , Kashiwadani Hideki , Sugimura Mitsutaka , Kuwaki Tomoyuki
Published: April 2021

Linalool odor exposure induces an analgesic effect in mice. This effect disappeared in the anosmic model mice, indicating that olfactory input evoked by linalool odor triggered this effect. Furthermore, hypothalamic orexinergic neurons play a pivotal role in this effect. However, the neuronal circuit mechanisms underlying this effect have not been fully addressed. In this study, we focused on the descending orexinergic projection to the spinal cord and examined whether this pathway contributes to the effect. We assessed the effect of intrathecal administration of orexin receptor antagonists on linalool odor-induced analgesia in the tail capsaicin test. We found that the selective orexin type 1 receptor antagonist, but not the selective orexin type 2 receptor antagonist, prevented the odor-induced analgesic effect. Furthermore, immunohistochemical analyses of c-Fos expression induced by the capsaicin test revealed that neuronal activity of spinal cord neurons was suppressed by linalool odor exposure, which was prevented by intrathecal administration of the orexin 1 receptor antagonist. These results indicate that linalool odor exposure drives the orexinergic descending pathway and suppresses nociceptive information flow at the spinal level.
Urinary neopterin of wild chimpanzees indicates that cell-mediated immune activity varies by age, sex, and female reproductive status
Authors: Negrey Jacob D. , Behringer Verena , Langergraber Kevin E. , Deschner Tobias
Published: April 2021

The study of free-living animal populations is necessary to understand life history trade-offs associated with immune investment. To investigate the role of life history strategies in shaping proinflammatory cell-mediated immune function, we analyzed age, sex, and reproductive status as predictors of urinary neopterin in 70 sexually mature chimpanzees (Pan troglodytes) at Ngogo, Kibale National Park, Uganda. In the absence of clinical signs of acute infectious disease, neopterin levels significantly increased with age in both male and female chimpanzees, as observed in humans and several other vertebrate species. Furthermore, males exhibited higher neopterin levels than females across adulthood. Finally, females with full sexual swellings, pregnant females, and post-reproductive females, the oldest individuals in our sample, exhibited higher neopterin levels than lactating females and cycling females without full swellings. Variation in females’ neopterin levels by reproductive status is consistent with post-ovulatory and pregnancy-related immune patterns documented in humans. Together, our results provide evidence of ample variation in chimpanzee immune activity corresponding to biodemographic and physiological variation. Future studies comparing immune activity across ecological conditions and social systems are essential for understanding the life histories of primates and other mammals.
Blue light blind-spot stimulation upregulates b-wave and pattern ERG activity in myopes
Authors: Amorim-de-Sousa Ana , Schilling Tim , Fernandes Paulo , Seshadri Yeshwanth , Bahmani Hamed , González-Méijome José Manuel
Published: April 2021

Upregulation of retinal dopaminergic activity may be a target treatment for myopia progression. This study aimed to explore the viability of inducing changes in retinal electrical activity with short-wavelength light targeting melanopsin-expressing retinal ganglion cells (ipRGCs) passing through the optic nerve head. Fifteen healthy non-myopic or myopic young adults were recruited and underwent stimulation with blue light using a virtual reality headset device. Amplitudes and implicit times from photopic 3.0 b-wave and pattern electroretinogram (PERG) were measured at baseline and 10 and 20 min after stimulation. Relative changes were compared between non-myopes and myopes. The ERG b-wave amplitude was significantly larger 20 min after blind-spot stimulation compared to baseline (p < 0.001) and 10 min (p < 0.001) post-stimulation. PERG amplitude P50-N95 also showed a significant main effect for ‘Time after stimulation’ (p < 0.050). Implicit times showed no differences following blind-spot stimulation. PERG and b-wave changes after blind-spot stimulation were stronger in myopes than non-myopes. It is possible to induce significant changes in retinal electrical activity by stimulating ipRGCs axons at the optic nerve head with blue light. The results suggest that the changes in retinal electrical activity are located at the inner plexiform layer and are likely to involve the dopaminergic system.
OxCOVID19 Database, a multimodal data repository for better understanding the global impact of COVID-19
Authors: Mahdi Adam , Błaszczyk Piotr , Dłotko Paweł , Salvi Dario , Chan Tak-Shing , Harvey John , Gurnari Davide , Wu Yue , Farhat Ahmad , Hellmer Niklas , Zarebski Alexander , Hogan Bernie , Tarassenko Lionel
Published: April 2021

Oxford COVID-19 Database (OxCOVID19 Database) is a comprehensive source of information related to the COVID-19 pandemic. This relational database contains time-series data on epidemiology, government responses, mobility, weather and more across time and space for all countries at the national level, and for more than 50 countries at the regional level. It is curated from a variety of (wherever available) official sources. Its purpose is to facilitate the analysis of the spread of SARS-CoV-2 virus and to assess the effects of non-pharmaceutical interventions to reduce the impact of the pandemic. Our database is a freely available, daily updated tool that provides unified and granular information across geographical regions.

Design type
Data integration objective

Measurement(s)
Coronavirus infectious disease, viral epidemiology

Technology type(s)
Digital curation

Factor types(s)

Sample characteristic(s)
Homo sapiens
Inhibition of glypican-1 expression induces an activated fibroblast phenotype in a human bone marrow-derived stromal cell-line
Authors: Kaur Sukhneeraj P. , Verma Arti , Lee Hee. K. , Barnett Lillie M. , Somanath Payaningal R. , Cummings Brian S.
Published: April 2021

Cancer-associated fibroblasts (CAFs) are the most abundant stromal cell type in the tumor microenvironment. CAFs orchestrate tumor-stromal interactions, and contribute to cancer cell growth, metastasis, extracellular matrix (ECM) remodeling, angiogenesis, immunomodulation, and chemoresistance. However, CAFs have not been successfully targeted for the treatment of cancer. The current study elucidates the significance of glypican-1 (GPC-1), a heparan sulfate proteoglycan, in regulating the activation of human bone marrow-derived stromal cells (BSCs) of fibroblast lineage (HS-5). GPC-1 inhibition changed HS-5 cellular and nuclear morphology, and increased cell migration and contractility. GPC-1 inhibition also increased pro-inflammatory signaling and CAF marker expression. GPC-1 induced an activated fibroblast phenotype when HS-5 cells were exposed to prostate cancer cell conditioned media (CCM). Further, treatment of human bone-derived prostate cancer cells (PC-3) with CCM from HS-5 cells exhibiting GPC-1 loss increased prostate cancer cell aggressiveness. Finally, GPC-1 was expressed in mouse tibia bone cells and present during bone loss induced by mouse prostate cancer cells in a murine prostate cancer bone model. These data demonstrate that GPC-1 partially regulates the intrinsic and extrinsic phenotype of human BSCs and transformation into activated fibroblasts, identify novel functions of GPC-1, and suggest that GPC-1 expression in BSCs exerts inhibitory paracrine effects on the prostate cancer cells. This supports the hypothesis that GPC-1 may be a novel pharmacological target for developing anti-CAF therapeutics to control cancer.
Surface modification of PMMA polymer and its composites with PC61BM fullerene derivative using an atmospheric pressure microwave argon plasma sheet
Authors: Sikora Andrzej , Czylkowski Dariusz , Hrycak Bartosz , Moczała-Dusanowska Magdalena , Łapiński Marcin , Dors Mirosław , Jasiński Mariusz
Published: April 2021

This paper presents the results of experimental investigations of the plasma surface modification of a poly(methyl methacrylate) (PMMA) polymer and PMMA composites with a [6,6]-phenyl-C61-butyric acid methyl ester fullerene derivative (PC61BM). An atmospheric pressure microwave (2.45 GHz) argon plasma sheet was used. The experimental parameters were: an argon (Ar) flow rate (up to 20 NL/min), microwave power (up to 530 W), number of plasma scans (up to 3) and, the kind of treated material. In order to assess the plasma effect, the possible changes in the wettability, roughness, chemical composition, and mechanical properties of the plasma-treated samples’ surfaces were evaluated by water contact angle goniometry (WCA), atomic force microscopy (AFM), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and X-ray photoelectron spectroscopy (XPS). The best result concerning the water contact angle reduction was from 83° to 29.7° for the PMMA material. The ageing studies of the PMMA plasma-modified surface showed long term (100 h) improved wettability. As a result of plasma treating, changes in the samples surface roughness parameters were observed, however their dependence on the number of plasma scans is irregular. The ATR-FTIR spectra of the PMMA plasma-treated surfaces showed only slight changes in comparison with the spectra of an untreated sample. The more significant differences were demonstrated by XPS measurements indicating the surface chemical composition changes after plasma treatment and revealing the oxygen to carbon ratio increase from 0.1 to 0.4.
Engineered adeno-associated virus 3 vector with reduced reactivity to serum antibodies
Authors: Ito Mika , Takino Naomi , Nomura Takamasa , Kan Akihiko , Muramatsu Shin-ichi
Published: April 2021

The natural serotypes of adeno-associated virus (AAV) or their variants, such as AAV8 and AAV5, are commonly used as vectors in the clinical programs for liver-targeted gene therapy. While AAV8 vectors are not highly efficient at targeting primary human hepatocytes, AAV3 vectors have recently demonstrated remarkable efficiency at targeting both human and non-human primate hepatocytes. However, the presence of high levels of neutralizing antibodies (NAbs) impedes transduction into hepatocytes, representing a major obstacle to the clinical application of AAV3 vectors. Herein, we engineered the viral capsid to reduce its reactivity with pre-existing NAbs, thereby enhancing the transduction efficiency. By introducing three substitutions (S472A, S587A, and N706A) on the surface loop of AAV3B capsid protein, we generated a triple mutant AAV3 (AAV.GT5) vector with less reactivity to anti-AAV capsid NAbs. While the transduction efficiency of AAV.GT5 into human hepatocellular cell lines was similar to those of parental AAV3B, it was 50-fold higher for hepatocytes derived from humanized mice compared to AAV8 vectors. Moreover, the AAV.GT5 vector yield was similar to those of the AAV2 and AAV3B vectors. Thus, high resistance to pre-existing NAbs makes AAV.GT5 a promising candidate for future liver-targeted gene therapy clinical trials.
Low serum neurofilament light chain values identify optimal responders to dimethyl fumarate in multiple sclerosis treatment
Authors: Walo-Delgado Paulette Esperanza , Sainz de la Maza Susana , Villarrubia Noelia , Monreal Enric , Medina Silvia , Espiño Mercedes , Fernández-Velasco José Ignacio , Rodríguez-Martín Eulalia , Roldán Ernesto , Lourido Daniel , Muriel Alfonso , Masjuan-Vallejo Jaime , Costa-Frossard Lucienne , Villar Luisa María
Published: April 2021

Serum neurofilament light chains (sNfL) are biomarkers of disease activity in multiple sclerosis (MS), but their value to predict response to treatment, and their association with patient immunological profile, need to be further explored. We studied 80 relapsing–remitting MS patients initiating dimethyl fumarate (DMF) treatment. sNfL levels were explored at baseline and at 3, 6 and 12 months by single molecule array. Blood lymphocyte subsets were measured at baseline and at 6 months by flow cytometry. Patients were followed a year and classified as NEDA (no evidence of disease activity) or ODA (ongoing disease activity). NEDA patients had lower sNfL levels at baseline (p = 0.0001), and after three (p = 0.004) and six (p = 0.03) months of DMF treatment. Consequently, low baseline sNfL values (≤ 12 pg/ml) increased the probability of NEDA (OR 5.8; CI 1.82–15.6; p = 0.002, after correcting by disease activity in the previous year), and associated with significant reductions of central memory CD4+ T lymphocytes, interferon-gamma+ CD8+ T lymphocytes, Natural Killer T cells, and memory B cells upon DMF treatment, being the highest differences in memory B cells (p < 0.0001). This shows that low baseline sNfL values identify MS patients with higher probability of optimal response to DMF and of a reduction in effector immune cells.
Estrogen withdrawal alters cytoskeletal and primary ciliary dynamics resulting in increased Hedgehog and osteoclastogenic paracrine signalling in osteocytes
Authors: Geoghegan Ivor P. , McNamara Laoise M. , Hoey David A.
Published: April 2021

Estrogen deficiency during post-menopausal osteoporosis leads to osteoclastogenesis and bone loss. Increased pro-osteoclastogenic signalling (RANKL/OPG) by osteocytes occurs following estrogen withdrawal (EW) and is associated with impaired focal adhesions (FAs) and a disrupted actin cytoskeleton. RANKL production is mediated by Hedgehog signalling in osteocytes, a signalling pathway associated with the primary cilium, and the ciliary structure is tightly coupled to the cytoskeleton. Therefore, the objective of this study was to investigate the role of the cilium and associated signalling in EW-mediated osteoclastogenic signalling in osteocytes. We report that EW leads to an elongation of the cilium and increase in Hedgehog and osteoclastogenic signalling. Significant trends were identified linking cilia elongation with reductions in cell area and % FA area/cell area, indicating that cilia elongation is associated with disruption of FAs and actin contractility. To verify this, we inhibited FA assembly via αvβ3 antagonism and inhibited actin contractility and demonstrated an elongated cilia and increased expression of Hh markers and Rankl expression. Therefore, our results suggest that the EW conditions associated with osteoporosis lead to a disorganisation of αvβ3 integrins and reduced actin contractility, which were associated with an elongation of the cilium, activation of the Hh pathway and osteoclastogenic paracrine signalling.
Impact of metformin treatment during pregnancy on maternal outcomes: a systematic review/meta-analysis
Authors: Tarry-Adkins Jane L. , Ozanne Susan E. , Aiken Catherine E.
Published: April 2021

We systematically assessed the impact of metformin treatment on maternal pregnancy outcomes. PubMed, Ovid Embase, Medline, Web of Science, ClinicalTrials.gov and Cochrane databases were systematically searched (inception-1st February 2021). Randomised controlled trials reporting pregnancy outcomes in women randomised to metformin versus any other treatment for any indication were included. Outcomes included gestational weight gain (GWG), pre-eclampsia, gestational hypertension, preterm birth, gestational age at delivery, caesarean section, gestational diabetes, glycaemic control, and gastrointestinal side-effects. Two independent reviewers conducted screening, with a third available to evaluate disagreements. Risk-of-bias and GRADE assessments were conducted using Cochrane Risk-of-Bias and GRADE-pro software. Thirty-five studies (n = 8033 pregnancies) met eligibility criteria. GWG was lower in pregnancies randomised to metformin versus other treatments (1.57 kg ± 0.60 kg; I2 = 86%, p < 0.0001), as was likelihood of pre-eclampsia (OR 0.69, 95% CI 0.50–0.95; I2 = 55%, p = 0.02). The risk of gastrointestinal side-effects was greater in metformin-exposed versus other treatment groups (OR 2.43, 95% CI 1.53–3.84; I2 = 76%, p = 0.0002). The risk of other maternal outcomes assessed was not significantly different between metformin-exposed versus other treatment groups. Metformin for any indication during pregnancy is associated with lower GWG and a modest reduced risk of pre-eclampsia, but increased gastrointestinal side-effects compared to other treatments.
Modeling and live imaging of mechanical instabilities in the zebrafish aorta during hematopoiesis
Authors: Chalin Dmitrii , Bureau Charlotte , Parmeggiani Andrea , Rochal Sergei , Kissa Karima , Golushko Ivan
Published: April 2021

All blood cells originate from hematopoietic stem/progenitor cells (HSPCs). HSPCs are formed from endothelial cells (ECs) of the dorsal aorta (DA), via endothelial-to-hematopoietic transition (EHT). The zebrafish is a primary model organism to study the process in vivo. While the role of mechanical stress in controlling gene expression promoting cell differentiation is actively investigated, mechanisms driving shape changes of the DA and individual ECs remain poorly understood. We address this problem by developing a new DA micromechanical model and applying it to experimental data on zebrafish morphogenesis. The model considers the DA as an isotropic tubular membrane subjected to hydrostatic blood pressure and axial stress. The DA evolution is described as a movement in the dimensionless controlling parameters space: normalized hydrostatic pressure and axial stress. We argue that HSPC production is accompanied by two mechanical instabilities arising in the system due to the plane stress in the DA walls and show how a complex interplay between mechanical forces in the system drives the emerging morphological changes.
N-Acetyl-l-leucine improves functional recovery and attenuates cortical cell death and neuroinflammation after traumatic brain injury in mice
Authors: Hegdekar Nivedita , Lipinski Marta M. , Sarkar Chinmoy
Published: April 2021

Traumatic brain injury (TBI) is a major cause of mortality and long-term disability around the world. Even mild to moderate TBI can lead to lifelong neurological impairment due to acute and progressive neurodegeneration and neuroinflammation induced by the injury. Thus, the discovery of novel treatments which can be used as early therapeutic interventions following TBI is essential to restrict neuronal cell death and neuroinflammation. We demonstrate that orally administered N-acetyl-l-leucine (NALL) significantly improved motor and cognitive outcomes in the injured mice, led to the attenuation of cell death, and reduced the expression of neuroinflammatory markers after controlled cortical impact (CCI) induced experimental TBI in mice. Our data indicate that partial restoration of autophagy flux mediated by NALL may account for the positive effect of treatment in the injured mouse brain. Taken together, our study indicates that treatment with NALL would be expected to improve neurological function after injury by restricting cortical cell death and neuroinflammation. Therefore, NALL is a promising novel, neuroprotective drug candidate for the treatment of TBI.
Stimulus dependence of interocular suppression
Authors: Lew Wei Hau , Stevenson Scott B. , Coates Daniel R.
Published: April 2021

Interocular suppression is the phenomenon in which the signal from one eye inhibits the other eye in the presence of dissimilar images. Various clinical and laboratory-based tests have been used to assess suppression, which vary in color, contrast, and stimulus size. These stimulus variations may yield different spatial extents of suppression, which makes it difficult to compare the outcomes. To evaluate the role of stimulus characteristics, we measured the suppression zone using a binocular rivalry paradigm in normally-sighted observers by systematically varying the stimulus parameters. The stimuli consist of a constantly visible horizontal reference seen by one eye while two vertical suppressors were presented to the other eye. With a keypress, the suppressors appeared for 1 s, to induce a transient suppression zone in the middle part of the reference. Subjects adjusted the width between the suppressors to determine the zone. The zone decreased significantly with increasing spatial frequency and lower contrast. The width was 1.4 times larger than the height. The zone was smaller with negative compared to positive contrast polarity but independent of eye dominance, luminance, and colored filters. A departure from scale invariance was captured with a model suggesting a stimulus-dependent and a small fixed non-stimulus-dependent portion.
Direct conversion of porcine primary fibroblasts into hepatocyte-like cells
Authors: Fráguas-Eggenschwiler Mariane , Eggenschwiler Reto , Söllner Jenny-Helena , Cortnumme Leon , Vondran Florian W. R. , Cantz Tobias , Ott Michael , Niemann Heiner
Published: April 2021

The pig is an important model organism for biomedical research, mainly due to its extensive genetic, physiological and anatomical similarities with humans. Until date, direct conversion of somatic cells into hepatocyte-like cells (iHeps) has only been achieved in rodents and human cells. Here, we employed lentiviral vectors to screen a panel of 12 hepatic transcription factors (TF) for their potential to convert porcine fibroblasts into hepatocyte-like cells. We demonstrate for the first time, hepatic conversion of porcine somatic cells by over-expression of CEBPα, FOXA1 and HNF4α2 (3TF-piHeps). Reprogrammed 3TF-piHeps display a hepatocyte-like morphology and show functional characteristics of hepatic cells, including albumin secretion, Dil-AcLDL uptake, storage of lipids and glycogen and activity of cytochrome P450 enzymes CYP1A2 and CYP2C33 (CYP2C9 in humans). Moreover, we show that markers of mature hepatocytes are highly expressed in 3TF-piHeps, while fibroblastic markers are reduced. We envision piHeps as useful cell sources for future studies on drug metabolism and toxicity as well as in vitro models for investigation of pig-to-human infectious diseases.
Apoptotic cells in mouse blastocysts are eliminated by neighbouring blastomeres
Authors: Pisko Jozef , Špirková Alexandra , Čikoš Štefan , Olexiková Lucia , Kovaříková Veronika , Šefčíková Zuzana , Fabian Dušan
Published: April 2021

Apoptosis is a physiological process that occurs commonly during the development of the preimplantation embryo. The present work examines the ability of apoptotic embryonic cells to express a signal promoting their phagocytosis, and quantifies the ability of neighbouring, normal embryonic cells to perform that task. Microscopic analysis of mouse blastocysts revealed phosphatidylserine externalization to be 10 times less common than incidence of apoptotic cells (as detected by TUNEL). In spite of the low frequency of phosphatidylserine-flipping (in inner cell mass, no annexin V staining was recorded), fluorescence staining of the plasma membrane showed more than 20% of apoptotic cells to have been engulfed by neighbouring blastomeres. The mean frequency of apoptotic cells escaping phagocytosis by their extrusion into blastocyst cavities did not exceed 10%. Immunochemically visualised RAC1 (an enzyme important in actin cytoskeleton rearrangement) was seen in phagosome-like structures containing a nucleus with a condensed morphology. Gene transcript analysis showed that the embryonic cells expressed 12 receptors likely involved in phagocytic process (Scarf1, Msr1, Cd36, Itgav, Itgb3, Cd14, Scarb1, Cd44, Stab1, Adgrb1, Cd300lf, Cd93). In conclusion, embryonic cells possess all the necessary mechanisms for recognising, engulfing and digesting apoptotic cells, ensuring the clearance of most dying blastomeres.
Cytocompatibility of stabilized black phosphorus nanosheets tailored by directly conjugated polymeric micelles for human breast cancer therapy
Authors: Biedulska M. , Jakóbczyk P. , Sosnowska M. , Dec B. , Muchlińska A. , Zaczek A. J. , Nidzworski D. , Bogdanowicz R.
Published: April 2021

The novel procedure of few-layer black phosphorus (FLBP) stabilization and functionalisation was here proposed. The cationic polymer PLL and non-ionic PEG have been involved into encapsulation of FLBP to allow sufficient time for further nanofabrication process and overcome environmental degradation. Two different spacer chemistry was designed to bind polymers to tumor-homing peptides. The efficiency of functionalisation was examined by RP-HPLC, microscopic (TEM and SEM) and spectroscopic (FT-IR and Raman) techniques as well supported by ab-initio modelling. The cell and dose dependent cytotoxicity of FLBP and its bioconjugates was evaluated against HB2, MCF-7 and MDA-MB-231 cell lines. Functionalisation allowed not only for improvement of environmental stability, but also enhances therapeutic effect by abolished the cytotoxicity of FLBP against HB2 cell line. Moreover, modification of FLBP with PLL caused increase of selectivity against highly aggressive breast cancer cell lines. Results indicate the future prospect application of black phosphorus nanosheets as nanocarrier, considering its unique features synergistically with conjugated polymeric micelles.