bluuebunny/crossref_metadata_2025 huggingface dataset:
A new role for RU486 (mifepristone): it protects sperm from premature capacitation during cryopreservation in buffalo
Authors: Dalal Jasmer , Kumar Pradeep , Chandolia R. K. , Pawaria Shikha , Rajendran Rasika , Sheoran Suman , Andonissamy Jerome , Kumar Dharmendra
Published: April 2019

The objective of this study was to determine the mechanism by which RU 486 (mifepristone) protects sperm to undergo premature capacitation during cryopreservation. For this, semen ejaculate (n = 20) was divided into four equal fractions and diluted using egg yolk-based extender supplemented with different concentrations of RU 486 (0, 5, 10 and 20 µM) and cryopreserved. We found that RU 486 did not impair the post-thaw sperm kinetics and motility but prevented cholesterol efflux, calcium influx, and protected CatSper channels during cryopreservation. The RU 486 protected sperm from premature capacitation which was confirmed by intracellular calcium level, expression of tyrosine phosphorylated proteins (75 and 80 kDa) and CTC (chlortetracycline) assay. Furthermore, antioxidant ability of RU 486 was reflected by the ferric reducing ability, lower production of sperm malondialdehyde and intracellular reactive oxygen species. Also, we demonstrated that RU 486 treated sperm underwent normal capacitation, zona pellucida binding and zygote cleavage indicating normal fertilizing ability of sperm. In conclusion, we report a new role of RU 486 in protecting buffalo sperm from premature capacitation during cryopreservation.
Fabrication of multifunctional titanium surfaces by producing hierarchical surface patterns using laser based ablation methods
Authors: Zwahr Christoph , Helbig Ralf , Werner Carsten , Lasagni Andrés Fabián
Published: April 2019

Textured implant surfaces with micrometer and sub-micrometer features can improve contact properties like cell adhesion and bacteria repellency. A critical point of these surfaces is their mechanical stability during implantation. Therefore, strategies capable to provide both biocompatibility for an improved implant healing and resistance to wear for protecting the functional surface are required. In this work, laser-based fabrication methods have been used to produce hierarchical patterns on titanium surfaces. Using Direct Laser Writing with a nanosecond pulsed laser, crater-like structures with a separation distance of 50 µm are produced on unpolished titanium surfaces. Directly on this texture, a hole-like pattern with 5 µm spatial period is generated using Direct Laser Interference Patterning with picosecond pulses. While the smaller features should reduce the bacterial adhesion, the larger geometry was designed to protect the smaller features from wear. On the multifunctional surface, the adherence of E. Coli bacteria is reduced by 30% compared to the untreated reference. In addition, wear test performed on the multiple-scale patterns demonstrated the possibility to protect the smaller features by the larger craters. Also, the influence of the laser treatment on the growth of a titanium oxide layer was evaluated using Energy Dispersive X-Ray Spectroscopy analysis.
Passive microinjection within high-throughput microfluidics for controlled actuation of droplets and cells
Authors: Azarmanesh Milad , Dejam Morteza , Azizian Pooya , Yesiloz Gurkan , Mohamad Abdulmajeed A. , Sanati-Nezhad Amir
Published: April 2019

Microinjection is an effective actuation technique used for precise delivery of molecules and cells into droplets or controlled delivery of genes, molecules, proteins, and viruses into single cells. Several microinjection techniques have been developed for actuating droplets and cells. However, they are still time-consuming, have shown limited success, and are not compatible with the needs of high-throughput (HT) serial microinjection. We present a new passive microinjection technique relying on pressure-driven fluid flow and pulsative flow patterns within an HT droplet microfluidic system to produce serial droplets and manage rapid and highly controlled microinjection into droplets. A microneedle is secured within the injection station to confine droplets during the microinjection. The confinement of droplets on the injection station prevents their movement or deformation during the injection process. Three-dimensional (3D) computational analysis is developed and validated to model the dynamics of multiphase flows during the emulsion generation. We investigate the influence of pulsative flows, microneedle parameters and synchronization on the efficacy of microinjection. Finally, the feasibility of implementing our microinjection model is examined experimentally. This technique can be used for tissue engineering, cells actuation and drug discovery as well as developing new strategies for drug delivery.
Toxicity and cytopathology mediated by Bacillus thuringiensis in the midgut of Anticarsia gemmatalis (Lepidoptera: Noctuidae)
Authors: Castro Bárbara Monteiro de Castro e , Martinez Luis Carlos , Barbosa Sergio Guedes , Serrão José Eduardo , Wilcken Carlos Frederico , Soares Marcus Alvarenga , da Silva Antonio Alberto , de Carvalho Amélia Guimarães , Zanuncio José Cola
Published: April 2019

Bioinsecticides and transgenic plants, based on Bacillus thuringiensis (Bt) toxins are important when managing Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae), a soybean defoliator pest. The interaction of these toxins with the caterpillar’s midgut cells determines their efficacy as an insecticide. The objective was to evaluate the toxicity of B. thuringiensis, subsp. kurstaki strain HD-1 and cytopathological changes mediated by these bacterial toxins in the midgut of A. gemmatalis caterpillars. Insecticidal efficacy was determined by calculating lethal concentration values (LC25, LC50, LC75, LC90 and LC99) in the laboratory. Midgut fragments from A. gemmatalis were extracted after bacterial ingestion and evaluated by light, transmission electron and confocal microscopy. The Bt median lethal concentrations showed toxicity [LC50 = 0.46 (0.43–0.49) mg mL−1] to fourth instar A. gemmatalis caterpillars after 108 hours. Bt induces severe cytotoxicity to A. gemmatalis midgut epithelial cells with increasing exposure over time, causing cellular disorganization, microvillus degeneration, cell fragmentation and protrusion, peritrophic membrane rupture, and cell vacuolization. The cell nuclei presented condensed chromatin and an increase in lysosome numbers. Apoptosis occurred in the midgut cells of caterpillars exposed to Bt. A regenerative response in A. gemmatalis caterpillars was observed 8 hours after exposure to Bt, however this response was not continuous. Toxins produced by Bt are harmful to A. gemmatalis at median concentration with structural damage and death of the midgut epithelial cells of this insect.
Structural analogues of roscovitine rescue the intracellular traffic and the function of ER-retained ABCB4 variants in cell models
Authors: Vauthier Virginie , Ben Saad Amel , Elie Jonathan , Oumata Nassima , Durand-Schneider Anne-Marie , Bruneau Alix , Delaunay Jean-Louis , Housset Chantal , Aït-Slimane Tounsia , Meijer Laurent , Falguières Thomas
Published: April 2019

Adenosine triphosphate binding cassette transporter, subfamily B member 4 (ABCB4) is the transporter of phosphatidylcholine at the canalicular membrane of hepatocytes. ABCB4 deficiency, due to genetic variations, is responsible for progressive familial intrahepatic cholestasis type 3 (PFIC3) and other rare biliary diseases. Roscovitine is a molecule in clinical trial that was shown to correct the F508del variant of cystic fibrosis transmembrane conductance regulator (CFTR), another ABC transporter. In the present study, we hypothesized that roscovitine could act as a corrector of ABCB4 traffic-defective variants. Using HEK and HepG2 cells, we showed that roscovitine corrected the traffic and localisation at the plasma membrane of ABCB4-I541F, a prototypical intracellularly retained variant. However, roscovitine caused cytotoxicity, which urged us to synthesize non-toxic structural analogues. Roscovitine analogues were able to correct the intracellular traffic of ABCB4-I541F in HepG2 cells. Importantly, the phospholipid secretion activity of this variant was substantially rescued by three analogues (MRT2-235, MRT2-237 and MRT2-243) in HEK cells. We showed that these analogues also triggered the rescue of intracellular traffic and function of two other intracellularly retained ABCB4 variants, i.e. I490T and L556R. Our results indicate that structural analogues of roscovitine can rescue genetic variations altering the intracellular traffic of ABCB4 and should be considered as therapeutic means for severe biliary diseases caused by this class of variations.
Efficacy and safety assessment of two enterococci phages in an in vitro biofilm wound model
Authors: Melo Luís D. R. , Ferreira R. , Costa Ana R. , Oliveira H. , Azeredo J.
Published: April 2019

Chronic wounds affect thousands of people worldwide, causing pain and discomfort to patients and represent significant economical burdens to health care systems. The treatment of chronic wounds is very difficult and complex, particularly when wounds are colonized by bacterial biofilms which are highly tolerant to antibiotics. Enterococcus faecium and Enterococcus faecalis are within the most frequent bacteria present in chronic wounds. Bacteriophages (phages) have been proposed as an efficient and alternative against antibiotic-resistant infections, as those found in chronic wounds. We have isolated and characterized two novel enterococci phages, the siphovirus vB_EfaS-Zip (Zip) and the podovirus vB_EfaP-Max (Max) to be applied during wound treatment. Both phages demonstrated lytic behavior against E. faecalis and E. faecium. Genome analysis of both phages suggests the absence of genes associated with lysogeny. A phage cocktail containing both phages was tested against biofilms formed in wound simulated conditions at a multiplicity of infection of 1.0 and a 2.5 log CFU.mL−1 reduction in the bacterial load after at 3 h of treatment was observed. Phages were also tested in epithelial cells colonized by these bacterial species and a 3 log CFU.mL−1 reduction was observed using both phages. The high efficacy of these new isolated phages against multi-species biofilms, their stability at different temperatures and pH ranges, short latent periods and non-cytotoxicity to epithelial cells suggest their therapeutic use to control infectious biofilms present in chronic wounds.
3D analysis of the whole subcutaneous adipose tissue reveals a complex spatial network of interconnected lobules with heterogeneous browning ability
Authors: Dichamp Jules , Barreau Corinne , Guissard Christophe , Carrière Audrey , Martinez Yves , Descombes Xavier , Pénicaud Luc , Rouquette Jacques , Casteilla Louis , Plouraboué Franck , Lorsignol Anne
Published: April 2019

Adipose tissue, as the main energy storage organ and through its endocrine activity, is interconnected with all physiological functions. It plays a fundamental role in energy homeostasis and in the development of metabolic disorders. Up to now, this tissue has been analysed as a pool of different cell types with very little attention paid to the organization and putative partitioning of cells. Considering the absence of a complete picture of the intimate architecture of this large soft tissue, we developed a method that combines tissue clearing, acquisition of autofluorescence or lectin signals by confocal microscopy, segmentation procedures based on contrast enhancement, and a new semi-automatic image analysis process, allowing accurate and quantitative characterization of the whole 3D fat pad organization. This approach revealed the unexpected anatomic complexity of the murine subcutaneous fat pad. Although the classical picture of adipose tissue corresponds to a superposition of simple and small ellipsoidal lobules of adipose cells separated by mesenchymal spans, our results show that segmented lobules display complex 3D poly-lobular shapes. Despite differences in shape and size, the number of these poly-lobular subunits is similar from one fat pad to another. Finally, investigation of the relationships of these subunits between each other revealed a never-described organization in two clusters with distinct molecular signatures and specific vascular and sympathetic nerve densities correlating with different browning abilities. This innovative procedure reveals that subcutaneous adipose tissue exhibits a subtle functional heterogeneity with partitioned areas, and opens new perspectives towards understanding its functioning and plasticity.
Effect of different cocoon stifling methods on the properties of silk fibroin biomaterials
Authors: Aznar-Cervantes Salvador D. , Pagan Ana , Monteagudo Santesteban Beatriz , Cenis José L.
Published: April 2019

Stifling treatments are applied to silk cocoons in order to kill the pupae, preventing the emergence of moths and allowing to preserve the silk during long periods of time. All of them involve the application of aggressive steps, such as sun exposure, hot steam from boiling water or hot air, during hours or even days. None of the scientific articles related to silk fibroin biomaterials has previously taken into account this fact in its section of materials and methods. In this work, the consequences of the stifling treatments most commonly used by the silk producing countries and companies are explored in depth, using fibroin films as biomaterial model. The protein degradation (visualised by SDS-PAGE) was dramatically increased in all the fibroin dissolutions produced from stifled cocoons; heavy and light chains of fibroin were specially degraded, reducing their presence along the lanes of the gel compared to the negative control (untreated fresh cocoons). Structural changes are also described for annealed silk fibroin films. The β-sheet content, analysed by means of infrared spectroscopy, was significantly higher when stifling was performed at higher temperature (70 °C and 85 °C). It is also exposed the impact of the stifling on the mechanical properties of the materials. Tensile strength and strain at break values were detected as significantly lower when this procedure was carried out by means of dry heat (85 °C) and sun exposure. On the other hand, and contrary to expectations, the proliferation of fibroblasts growing on the materials was improved by all the different stifling methods, compared to negative control, being this improvement, especially accentuated, on the films produced with fibroin purified from cocoons treated with dry heat.
CRISPR-Cas9 Editing in Maize: Systematic Evaluation of Off-target Activity and Its Relevance in Crop Improvement
Authors: Young Joshua , Zastrow-Hayes Gina , Deschamps Stéphane , Svitashev Sergei , Zaremba Mindaugas , Acharya Ananta , Paulraj Sushmitha , Peterson-Burch Brooke , Schwartz Chris , Djukanovic Vesna , Lenderts Brian , Feigenbutz Lanie , Wang Lijuan , Alarcon Clara , Siksnys Virginijus , May Gregory , Chilcoat N. Doane , Kumar Sandeep
Published: April 2019

CRISPR-Cas9 enabled genome engineering has great potential for improving agriculture productivity, but the possibility of unintended off-target edits has evoked some concerns. Here we employ a three-step strategy to investigate Cas9 nuclease specificity in a complex plant genome. Our approach pairs computational prediction with genome-wide biochemical off-target detection followed by validation in maize plants. Our results reveal high frequency (up to 90%) on-target editing with no evidence of off-target cleavage activity when guide RNAs were bioinformatically predicted to be specific. Predictable off-target edits were observed but only with a promiscuous guide RNA intentionally designed to validate our approach. Off-target editing can be minimized by designing guide RNAs that are different from other genomic locations by at least three mismatches in combination with at least one mismatch occurring in the PAM proximal region. With well-designed guides, genetic variation from Cas9 off-target cleavage in plants is negligible, and much less than inherent variation.
Comparative analysis of serologic cross-reactivity using convalescent sera from filovirus-experimentally infected fruit bats
Authors: Schuh Amy J. , Amman Brian R. , Sealy Tara S. , Flietstra Timothy D. , Guito Jonathan C. , Nichol Stuart T. , Towner Jonathan S.
Published: April 2019

With the exception of Reston and Bombali viruses, the marburgviruses and ebolaviruses (family Filoviridae) cause outbreaks of viral hemorrhagic fever in sub-Saharan Africa. The Egyptian rousette bat (ERB) is a natural reservoir host for the marburgviruses and evidence suggests that bats are also natural reservoirs for the ebolaviruses. Although the search for the natural reservoirs of the ebolaviruses has largely involved serosurveillance of the bat population, there are no validated serological assays to screen bat sera for ebolavirus-specific IgG antibodies. Here, we generate filovirus-specific antisera by prime-boost immunization of groups of captive ERBs with all seven known culturable filoviruses. After validating a system of filovirus-specific indirect ELISAs utilizing infectious-based virus antigens for detection of virus-specific IgG antibodies from bat sera, we assess the level of serological cross-reactivity between the virus-specific antisera and heterologous filovirus antigens. This data is then used to generate a filovirus antibody fingerprint that can predict which of the filovirus species in the system is most antigenically similar to the species responsible for past infection. Our filovirus IgG indirect ELISA system will be a critical tool for identifying bat species with high ebolavirus seroprevalence rates to target for longitudinal studies aimed at establishing natural reservoir host-ebolavirus relationships.
An HRM Assay to Differentiate Sheeppox Virus Vaccine Strains from Sheeppox Virus Field Isolates and other Capripoxvirus Species
Authors: Chibssa Tesfaye Rufael , Settypalli Tirumala Bharani K. , Berguido Francisco J. , Grabherr Reingard , Loitsch Angelika , Tuppurainen Eeva , Nwankpa Nick , Tounkara Karim , Madani Hafsa , Omani Amel , Diop Mariane , Cattoli Giovanni , Diallo Adama , Lamien Charles Euloge
Published: April 2019

Sheep poxvirus (SPPV), goat poxvirus (GTPV) and lumpy skin disease virus (LSDV) affect small ruminants and cattle causing sheeppox (SPP), goatpox (GTP) and lumpy skin disease (LSD) respectively. In endemic areas, vaccination with live attenuated vaccines derived from SPPV, GTPV or LSDV provides protection from SPP and GTP. As live poxviruses may cause adverse reactions in vaccinated animals, it is imperative to develop new diagnostic tools for the differentiation of SPPV field strains from attenuated vaccine strains. Within the capripoxvirus (CaPV) homolog of the variola virus B22R gene, we identified a unique region in SPPV vaccines with two deletions of 21 and 27 nucleotides and developed a High-Resolution Melting (HRM)-based assay. The HRM assay produces four distinct melting peaks, enabling the differentiation between SPPV vaccines, SPPV field isolates, GTPV and LSDV. This HRM assay is sensitive, specific, and provides a cost-effective means for the detection and classification of CaPVs and the differentiation of SPPV vaccines from SPPV field isolates.
Chemogenetic stimulation of the infralimbic cortex reverses alcohol-induced fear memory overgeneralization
Authors: Scarlata M. J. , Lee S. H. , Lee D. , Kandigian S. E. , Hiller A. J. , Dishart J. G. , Mintz G. E. , Wang Z. , Coste G. I. , Mousley A. L. , Soler I. , Lawson K. , Ng A. J. , Bezek J. L. , Bergstrom H. C.
Published: April 2019

Post-traumatic stress disorder (PTSD) and alcohol use disorder (AUD) are often comorbid. Drinking tends to increase following trauma, which may exacerbate PTSD-related symptoms. Despite a clear relationship between excessive alcohol use and PTSD, how alcohol impacts the expression of traumatic fear remains unclear. This study aims to determine the neurobehavioral impact of chronic alcohol (ethanol; EtOH) on the expression of established fear memories in C57BL/6 N mice. We show that chronic EtOH selectively augments cued fear memory generalization and impairs fear extinction retrieval, leaving the expression of the original cued response intact. Immunohistochemistry for Arc/arg3.1 (Arc) revealed EtOH-induced decreases in Arc expression in the infralimbic cortex (IL) and basolateral amygdala complex (BLA) that were associated with cued fear memory overgeneralization. Chemogenetic stimulation of IL pyramidal neurons reversed EtOH-driven fear memory overgeneralization, identifying a role for the IL in cued fear memory precision. Considering the modulatory influence of the IL over conditioned fear expression, these data suggest a model whereby chronic EtOH-driven neuroadaptations in the IL promote fear memory overgeneralization. These findings provide new mechanistic insight into how excessive alcohol use, following exposure to a traumatic event, can exacerbate symptoms of traumatic fear.
Susceptibility to infection with Borrelia afzelii and TLR2 polymorphism in a wild reservoir host
Authors: Gomez-Chamorro Andrea , Battilotti Florian , Cayol Claire , Mappes Tapio , Koskela Esa , Boulanger Nathalie , Genné Dolores , Sarr Anouk , Voordouw Maarten Jeroen
Published: April 2019

The study of polymorphic immune genes in host populations is critical for understanding genetic variation in susceptibility to pathogens. Controlled infection experiments are necessary to separate variation in the probability of exposure from genetic variation in susceptibility to infection, but such experiments are rare for wild vertebrate reservoir hosts and their zoonotic pathogens. The bank vole (Myodes glareolus) is an important reservoir host ofBorrelia afzelii, a tick-borne spirochete that causes Lyme disease. Bank vole populations are polymorphic for Toll-like receptor 2 (TLR2), an innate immune receptor that recognizes bacterial lipoproteins. To test whether the TLR2 polymorphism influences variation in the susceptibility to infection withB.afzelii, we challenged pathogen-free, lab-born individuals of known TLR2 genotype withB.afzelii-infected ticks. We measured the spirochete load in tissues of the bank voles. The susceptibility to infection withB.afzeliifollowing an infected tick bite was very high (95%) and did not differ between TLR2 genotypes. The TLR2 polymorphism also had no effect on the spirochete abundance in the tissues of the bank voles. Under the laboratory conditions of our study, we did not find that the TLR2 polymorphism in bank voles influenced variation in the susceptibility toB.afzeliiinfection.
Genetic analysis reveals unprecedented diversity of a globally-important plant pathogenic genus
Authors: Garfinkel Andrea R. , Coats Katie P. , Sherry Don L. , Chastagner Gary A.
Published: April 2019

Genus Botrytis contains approximately 35 species, many of which are economically-important and globally-distributed plant pathogens which collectively infect over 1,400 plant species. Recent efforts to genetically characterize genus Botrytis have revealed new species on diverse host crops around the world. In this study, surveys and subsequent genetic analysis of the glyceraldehyde-3-phosate dehydrogenase (G3PDH), heat-shock protein 60 (HSP60), DNA-dependent RNA polymerase subunit II (RPB2), and necrosis and ethylene-inducing proteins 1 and 2 (NEP1 and NEP2) genes indicated that Botrytis isolates collected from peony fields in the United States contained more species diversity than ever before reported on a single host, including up to 10 potentially novel species. Together, up to 16 different phylogenetic species were found in association with peonies in the Pacific Northwest, which is over a third of the total number of species that are currently named. Furthermore, species were found on peonies in Alaska that have been described on other host plants in different parts of the world, indicating a wider geographic and host distribution than previously thought. Lastly, some isolates found on peony share sequence similarity with unnamed species found living as endophytes in weedy hosts, suggesting that the isolates found on peony have flexible lifestyles as recently discovered in the genus. Selected pathogenicity, growth, and morphological characteristics of the putatively new Botrytis species were also assessed to provide a basis for future formal description of the isolates as new species.
Factors associated with chronic obstructive pulmonary disease exacerbation, based on big data analysis
Authors: Lee Jongmin , Jung Hyun Myung , Kim Sook Kyung , Yoo Kwang Ha , Jung Ki-Suck , Lee Sang Haak , Rhee Chin Kook
Published: April 2019

Preventing exacerbation in chronic obstructive pulmonary disease (COPD) patients is crucial, but requires identification of the exacerbating factors. To date, no integrated analysis of patient-derived and external factors has been reported. To identify factors associated with COPD exacerbation, we collected data, including smoking status, lung function, and COPD assessment test scores, from 594 COPD patients in the Korean COPD subgroup study (KOCOSS), and merged these data with patients’ Korean Health Insurance Review and Assessment Service data for 2007–2012. We also collected primary weather variables, including levels of particulate matter <10 microns in diameter, daily minimum ambient temperature, as well as respiratory virus activities, and the logs of web queries on COPD-related issues. We then assessed the associations between these patient-derived and external factors and COPD exacerbations. Univariate analysis showed that patient factors, air pollution, various types of viruses, temperature, and the number of COPD-related web queries were associated with COPD exacerbation. Multivariate analysis revealed that the number of exacerbations in the preceding year, female sex, COPD grade, and influenza virus detection rate, and lowest temperature showed significant association with exacerbation. Our findings may help COPD patients predict when exacerbations are likely, and provide intervention as early as possible.
A comparative study of energy and carbon efficiency for emerging countries using panel stochastic frontier analysis
Authors: Jin Taeyoung , Kim Jinsoo
Published: April 2019

The demand for energy has been increasing significantly worldwide. Consequently, carbon emissions have accelerated, since energy usage involves carbon dioxide. Given that the available energy has quantitative restriction feature, efficient usage of energy becomes crucial. Energy efficiency is expected to improve over time with technological advancements. However, the adoption of low-carbon energy technology caused by the growing concern about carbon emissions may actually offset energy efficiency, owing to the higher cost compared with traditional energy methods. We conducted a stochastic frontier analysis to examine energy efficiency in the views of both economic and ecological aspect view during 1995–2016 for 21 emerging countries selected from Morgan Stanley Capital International, using energy consumption, economic complexity index and the other factors of production based on the Cobb-Douglas production function. Mexico was identified as one of the most energy-efficient countries; however, Mexico could not be classified as real energy efficient, as it demonstrated the highest carbon inefficiency level. We also categorized countries demonstrating increased economic energy efficiency and decreased carbon inefficiency as frontier country, and identified six such countries.
Propagation and Selectivity of Axonal Loss in Leber Hereditary Optic Neuropathy
Authors: Coussa Razek Georges , Merat Pooya , Levin Leonard A.
Published: April 2019

Leber hereditary optic neuropathy (LHON) is a syndrome of subacute loss of central vision associated with mutations in mitochondrial DNA coding for components of complex I. LHON preferentially involves small axons in the temporal optic nerve, but the reason is unclear. We performed a Monte Carlo simulation of the spread of injury in LHON axons to better understand the predilection for small axons. Optic nerve slices were modeled as grids containing axons with sizes from reported regional distributions. The propagation of injury from a localized concentration of superoxide was simulated as the spread via passive diffusion from one axon to adjacent axons, with basal production and scavenging rate proportional to axonal area and volume, respectively. Axonal degeneration occurred when intra-axonal concentrations reached a toxic threshold. Simulations demonstrated that almost all small and medium axons degenerated by the time steady-state was reached, but about 50% of large axons were preserved. The location of initial injury affected time to steady state, with nasal injuries reaching steady state faster than temporal injuries. The pattern of axonal degeneration in the simulations mirrored both visual fields and optic nerve histology from patients with LHON. These results provide insight into the nature of axonal loss in LHON.
Development of a novel cultivation technique for uncultured soil bacteria
Authors: Chaudhary Dhiraj Kumar , Khulan Altankhuu , Kim Jaisoo
Published: April 2019

In this study, a new diffusion bioreactor was developed to cultivate hidden bacterial communities in their natural environment. The newly developed method was investigated to cultivate microbial communities from the forest soil, and the results were evaluated against traditional culture methods and compared to the results of a pyrosequencing-based molecular survey. The molecular analysis revealed that a diverse bacterial population was present in the soil sample. However, both the newly developed method and the traditional method recovered more than 400 isolates, which belonged to only four phyla: Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes. Although these isolates were distributed over only four major phyla, the use of the newly developed technique resulted in the successful cultivation of 35 previously uncultured strains, whereas no such strains were successfully cultivated by the traditional method. Furthermore, the study also found that the recovery of uncultured bacteria and novel isolates was related to sampling season, incubation period, and cultivation media. The use of soil collected in summer, a prolonged incubation period, and low-substrate modified media increased the recovery of uncultured and novel isolates. Overall, the results indicate that the newly designed diffusion bioreactor can mimic the natural environment, which permits the cultivation of previously uncultured bacteria.
Reduced caveolae density in arteries of SHR contributes to endothelial dysfunction and ROS production
Authors: Potje Simone R. , Grando Marcella D. , Chignalia Andreia Z. , Antoniali Cristina , Bendhack Lusiane M.
Published: April 2019

Caveolae are plasma membrane invaginations enriched with high cholesterol and sphingolipid content; they also contain caveolin proteins in their structure. Endothelial nitric oxide synthase (eNOS), an enzyme that synthesizes nitric oxide (NO) by converting L-arginine to L-citrulline, is highly concentrated in plasma membrane caveolae. Hypertension is associated with decreased NO production and impaired endothelium-dependent relaxation. Understanding the molecular mechanisms that follow hypertension is important. For this study, we hypothesized that spontaneously hypertensive rat (SHR) vessels should have a smaller number of caveolae, and that the caveolae structure should be disrupted in these vessels. This should impair the eNOS function and diminish NO bioavailability. Therefore, we aimed to investigate caveolae integrity and density in SHR aortas and mesenteric arteries and the role played by caveolae in endothelium-dependent relaxation. We have been able to show the presence of caveolae-like structures in SHR aortas and mesenteric arteries. Increased phenylephrine-induced contractile response after treatment with dextrin was related to lower NO release. In addition, impaired acetylcholine-induced endothelium-dependent relaxation could be related to decreased caveolae density in SHR vessels. The most important finding of this study was that cholesterol depletion with dextrin induced eNOS phosphorylation at Serine1177 (Ser1177) and boosted reactive oxygen species (ROS) production in normotensive rat and SHR vessels, which suggested eNOS uncoupling. Dextrin plus L-NAME or BH4 decreased ROS production in aorta and mesenteric arteries supernatant’s of both SHR and normotensive groups. Human umbilical vein endothelial cells (HUVECs) treated with dextrin confirmed eNOS uncoupling, as verified by the reduced eNOS dimer/monomer ratio. BH4, L-arginine, or BH4 plus L-arginine inhibited eNOS monomerization. All these results showed that caveolae structure and integrity are essential for endothelium-dependent relaxation. Additionally, a smaller number of caveolae is associated with hypertension. Finally, caveolae disruption promotes eNOS uncoupling in normotensive and hypertensive rat vessels and in HUVECs.
The utility of STOP-BANG questionnaire in the sleep-lab setting
Authors: Kuczyński Wojciech , Mokros Łukasz , Stolarz Aleksandra , Białasiewicz Piotr
Published: April 2019

Polysomnography (PSG) is considered the gold standard in obstructive sleep apnea-hypopnea syndrome (OSAS) diagnostics, but its availability is still limited. Thus, it seems useful to assess patients pre-diagnostic risk for OSAS to prioritize the use of this examination. The purpose of this study was to assess positive (PPV) and negative (NPV) predictive values of the STOP BANG questionnaire (SBQ) in patients with presumptive diagnosis of OSAS. From a database of 1,171 (880 men) patients of a university based sleep center, 1,123 (847 men) met eligibility criteria and their SBQ scores were subject to the Bayesian analysis. The analysis of PPV and NPV was conducted at all values of SBQ for all subjects, but also separately for males and females, and for total sleep time (TS) and for sleep in the lateral position (LP). The probability of OSAS (AHI ≥ 5) and at least moderate OSAS (AHI ≥ 15) for TS was 0.766 and 0.516, while for LP the values were 0.432 and 0.289, respectively. Overall, due to low specificity, SBQ had low PPV for TS and LP. Negative test result (SBQ < 3) revealed NPV of 0.620 at AHI < 5 and 0.859 at AHI < 15 for TS, while in LP NPV values were 0.935 at AHI < 5 and 1.0 at AHI < 15, (n = 31), while SBQ < 4 generated NPV of 0.943 in LP (n = 105). SBQ did not change probabilities of OSAS to confirm or rebut diagnosis for TS. However, it is highly probable that SQB can rule out OSAS diagnosis at AHI ≥ 15 for LP.
The Late Effects of Radiation Therapy on Skeletal Muscle Morphology and Progenitor Cell Content are Influenced by Diet-Induced Obesity and Exercise Training in Male Mice
Authors: D’Souza Donna , Roubos Sophia , Larkin Jillian , Lloyd Jessica , Emmons Russell , Chen Hong , De Lisio Michael
Published: April 2019

Radiation exposure during muscle development induces long-term decrements to skeletal muscle health, which contribute to reduced quality of life in childhood cancer survivors. Whether the effects of radiation on skeletal muscle are influenced by relevant physiological factors, such as obesity and exercise training remains unknown. Using skeletal muscle from our previously published work examining the effects of obesity and exercise training on radiation-exposed bone marrow, we evaluated the influence of these physiological host factors on irradiated skeletal muscle morphology and cellular dynamics. Mice were divided into control and high fat diet groups with or without exercise training. All mice were then exposed to radiation and continued in their intervention group for an additional 4 weeks. Diet-induced obesity resulted in increased muscle fibrosis, while obesity and exercise training both increased muscle adiposity. Exercise training enhanced myofibre cross-sectional area and the number of satellite cells committed to the myogenic lineage. High fat groups demonstrated an increase in p-NFĸB expression, a trend for a decline in IL-6, and increase in TGFB1. These findings suggest exercise training improves muscle morphology and satellite cell dynamics compared to diet-induced obesity in irradiated muscle, and have implications for exercise interventions in cancer survivors.
Age-Dependent Variation of Lamina Cribrosa Displacement During the Standardized Valsalva Maneuver
Authors: Kim Yong Woo , Lee Dong Hyun , Lim Hyung Bin , Oh Baek-Lok , Kim Young Kook , Girard Michael J. A. , Mari Jean Martial , Park Ki Ho , Jeoung Jin Wook
Published: April 2019

Based on biomechanical theory, lamina cribrosa (LC) displacement, the key component of progressive glaucomatous change, is presumed to be dependent on intraocular pressure (IOP) as well as tissue stiffness of LC. In the performance of the Valsalva maneuver, both IOP and cerebrospinal fluid pressure can increase. The present study investigated the age-dependent variation of LC displacement during the standardized Valsalva maneuver in healthy subjects. Sixty-three (63) eyes (age range: 20–76 years) were prospectively underwent IOP measurement and Cirrus HD-OCT optic disc scans before and during the standardized Valsalva maneuver. During the standardized Valsalva maneuver, the IOP significantly increased from 13.2 ± 2.9 mmHg to 18.6 ± 5.2 mmHg (P < 0.001). The maximal LC depth significantly decreased in the younger age groups (age: 20 s to 40 s) but not in the older age groups (age: over 50). The BMO distance did not change significantly. Younger age (P = 0.009), a smaller increase of IOP during the Valsalva maneuver (P = 0.002), and greater baseline maximal LC depth (P = 0.013) were associated with more anterior displacement of the LC during the standardized Valsalva maneuver. Taken together, age as well as translaminar pressure dynamics seems to play a crucial role in LC biomechanics.
Violent paroxysmal activity drives self-feeding magma replenishment at Mt. Etna
Authors: Viccaro Marco , Giuffrida Marisa , Zuccarello Francesco , Scandura Mariabenedetta , Palano Mimmo , Gresta Stefano
Published: April 2019

A new sequence of eruptions occurred at Mt. Etna volcano during the first half of 2017, after almost 8 months of quiescence. These episodes had low-to-mild intensity and markedly differ from the violent paroxysms occurred at the Voragine Crater (VOR) during December 2015 and May 2016. Despite the general weak explosive nature of the eruptions, the activity during 2017 revealed unusually complex dynamics of magma ascent and interaction. Detection and investigation of such dynamics required a multidisciplinary approach in which bulk rock compositions, crystal chemical zoning, diffusion chronometry and ground deformation data have been combined. Bulk rock major and trace elements suggest that the 2017 magmas followed a differentiation path similar to that experienced by magmas erupted at Mt. Etna during the 2015–16 eruptions at VOR. Olivine core compositions and zoning patterns indicate the presence of multiple magmatic environments at depth that strictly interacted each other through some episodes of intrusion and mixing before and during the 2017 eruptive events. Timescales retrieved from diffusion chronometry on olivine normal and reverse zoning correlate well with the ground deformation stages detected through geodetic data and associated models, thus allowing to track the evolution through time of the 2017 volcanic activity. Combination of all petrological and geodetic observations supports the idea that dynamics of magma transfer driving the eruptive episodes of 2017 have been a direct consequence of the violent eruptions occurred at VOR on May 2016, which boosted the ascent of new magma from depth and improved the efficiency of the plumbing system to transfer it upward to the surface. We propose a mechanism of self-feeding replenishment of the volcano plumbing system during 2017, where magma recharge from depth is triggered by sudden unloading of the magma column consequential to the violent paroxysmal activity occurred on May 2016 at VOR.
Non-uniformity smoothing of direct-driven fuel target implosion by phase control in heavy ion inertial fusion
Authors: Sato R. , Kawata S. , Karino T. , Uchibori K. , Ogoyski A. I.
Published: April 2019

We have proposed a dynamic smoothing method based on a phase control to smooth plasma non-uniformities in perturbed plasma systems. In this paper, the dynamic smoothing method is applied to a spherical direct-driven fuel target implosion in heavy ion inertial confinement fusion. We found that the wobbling motion of each heavy ion beam (HIB) axis induces a phase-controlled HIBs energy deposition, and consequently the phase-controlled implosion acceleration is realized, so that the HIBs irradiation non-uniformity is successfully smoothed. HIB accelerators provide a well-established performance to oscillate a HIB axis at a high frequency. In inertial confinement fusion, a fuel implosion uniformity is essentially significant for achieving the DT fuel compression and for releasing the fusion energy, and the non-uniformity of the implosion acceleration should be less than a few %. The results in this paper demonstrate that the wobbling HIBs would provide an improvement in the fuel target implosion uniformity.
A novel multidomain acyl-CoA carboxylase in Saccharopolyspora erythraea provides malonyl-CoA for de novo fatty acid biosynthesis
Authors: Livieri Andrea L. , Navone Laura , Marcellin Esteban , Gramajo Hugo , Rodriguez Eduardo
Published: April 2019

Acetyl-CoA carboxylases (ACCs) are enzyme complexes generally composed of three catalytic domains and distributed in all organisms. In prokaryotes and plastids of most plants, these domains are encoded in distinct subunits forming heteromeric complexes. Distinctively, cytosolic ACCs from eukaryotes and plastids of graminaceous monocots, are organized in a single multidomain polypeptide. Until now, no multidomain ACCs had been discovered in bacteria. Here, we show that a putative multidomain ACC in Saccharopolyspora erythraea is encoded by the sace_4237 gene, representing the first prokaryotic ACC homodimeric multidomain complex described. The SACE_4237 complex has both acetyl-CoA and propionyl-CoA carboxylase activities. Importantly, we demonstrate that sace_4237 is essential for S. erythraea survival as determined by the construction of a sace_4237 conditional mutant. Altogether, our results show that this prokaryotic homodimeric multidomain ACC provides malonyl-CoA for de novo fatty acid biosynthesis. Furthermore, the data presented here suggests that evolution of these enzyme complexes, from single domain subunits to eukaryotic multidomain ACCs, occurred in bacteria through domain fusion.
High levels of drug resistance in commensal E. coli in a cohort of children from rural central India
Authors: Purohit Manju Raj , Lindahl Lars Falkdalen , Diwan Vishal , Marrone Gaetano , Lundborg Cecilia Stålsby
Published: April 2019

The world is experiencing crisis of antibiotic resistance not only in pathogenic but also in commensal bacteria. We determine the prevalence of antibiotic resistance in commensal Escherichia coli in young children in rural setting of central India and search for its correlations with demographic and behavioral factors. At seven time points during a period of 2 years we collected stool samples from 125 children; aged 1–3 in a rural area of Madhya Pradesh. We isolated six isolates of E. coli per stool sample and subjected them to antibiotic susceptibility testing. We found resistance to ampicillin, quinolones, cephalosporins, sulfamethoxazole, co-trimoxazole, in at least one isolate from 89% to 100% of children. Extended spectrum beta lactamase producing E. coli were identified in all but one child and multidrug resistance was identified in isolates from all children. Female gender (p = 0.04) and higher wealth (p = 0.03) was significantly correlated with less antibiotic resistance. Thus, the high prevalence of antibiotic resistance in commensal E. coli in rural community from India needs urgent measures to control the growing antibiotic resistance crisis.
A transnational perspective of global and regional ecosystem service flows from and to mountain regions
Authors: Schirpke Uta , Tappeiner Ulrike , Tasser Erich
Published: April 2019

The spatial relationships of ecosystem services are complex and poorly understood due to spatial mismatches between areas of provision and the areas that benefit. In this study, we assess the spatial flows of six key ecosystem services from and to mountain regions at the regional and global level. We identify major directions of spatial flow and illustrate different types and transfer mechanisms with detailed examples focusing on the European Alps and surrounding lowlands. Our results demonstrate that the spatial flows of ecosystem services range from local to global interactions and extend far beyond the regional level for most of the ecosystem services assessed. Transportation processes encompass passive biophysical processes and the active transportation of goods, distribution of information and traveling of people. Decision and policy-making can use this enhanced understanding to influence ecosystem service transfer and consequently manage natural resources in a sustainable way.
Role of Oxide/Metal Bilayer Electrodes in Solution Processed Organic Field Effect Transistors
Authors: Ablat Abduleziz , Kyndiah Adrica , Houin Geoffroy , Alic Tugbahan Yilmaz , Hirsch Lionel , Abbas Mamatimin
Published: April 2019

High performance, air stable and solution-processed small molecule 2,7-dioctyl[1]benzothieno[3,2-b]benzothiophene (C8-BTBT) based organic field-effect transistors (OFETs) with various electrode configurations were studied in detail. The contact resistance of OFET devices with Ag, Au, WO3/Ag, MoO3/Ag, WO3/Au, and MoO3/Au were compared. Reduced contact resistance and consequently improved performance were observed in OFET devices with oxide interlayers compared to the devices with bare metal electrodes. The best oxide/metal combination was determined. The possible mechanisms for enhanced electrical properties were explained by favorable morphological and electronic structure of organic/metal oxide/metal interfaces.
Reduced Basal Nitric Oxide Production Induces Precancerous Mammary Lesions via ERBB2 and TGFβ
Authors: Ren Gang , Zheng Xunzhen , Bommarito Matthew , Metzger Samantha , Walia Yashna , Letson Joshua , Schroering Allen , Kalinoski Andrea , Weaver David , Figy Christopher , Yeung Kam , Furuta Saori
Published: April 2019

One third of newly diagnosed breast cancers in the US are early-stage lesions. The etiological understanding and treatment of these lesions have become major clinical challenges. Because breast cancer risk factors are often linked to aberrant nitric oxide (NO) production, we hypothesized that abnormal NO levels might contribute to the formation of early-stage breast lesions. We recently reported that the basal level of NO in the normal breast epithelia plays crucial roles in tissue homeostasis, whereas its reduction contributes to the malignant phenotype of cancer cells. Here, we show that the basal level of NO in breast cells plummets during cancer progression due to reduction of the NO synthase cofactor, BH4, under oxidative stress. Importantly, pharmacological deprivation of NO in prepubertal to pubertal animals stiffens the extracellular matrix and induces precancerous lesions in the mammary tissues. These lesions overexpress a fibrogenic cytokine, TGFβ, and an oncogene, ERBB2, accompanied by the occurrence of senescence and stem cell-like phenotype. Consistently, normalization of NO levels in precancerous and cancerous breast cells downmodulates TGFβ and ERBB2 and ameliorates their proliferative phenotype. This study sheds new light on the etiological basis of precancerous breast lesions and their potential prevention by manipulating the basal NO level.