bluuebunny/crossref_metadata_2025 huggingface dataset:
Small Molecules Attenuate the Interplay between Conformational Fluctuations, Early Oligomerization and Amyloidosis of Alpha Synuclein
Authors: Ghosh Sumanta , Kundu Amrita , Chattopadhyay Krishnananda
Published: March 2018

Aggregation of alpha synuclein has strong implications in Parkinson’s disease. The heterogeneity of folding/aggregation landscape and transient nature of the early intermediates result in difficulty in developing a successful therapeutic intervention. Here we used fluorescence measurements at ensemble and single molecule resolution to study how the late and early events of alpha synuclein aggregation modulate each other. In-vitro aggregation data was complemented using measurements inside live neuroblastoma cells by employing a small molecule labeling technique. An inhibitor molecule (arginine), which delayed the late event of amyloidosis, was found to bind to the protein, shifting the early conformational fluctuations towards a compact state. In contrast, a facilitator of late aggregation (glutamate), was found to be excluded from the protein surface. The presence of glutamate was found to speed up the oligomer formation at the early stage. We found that the effects of the inhibitor and facilitator were additive and as a result they maintained a ratio at which they cancelled each other’s influence on different stages of alpha synuclein aggregation.
Functional cyclophilin D moderates platelet adhesion, but enhances the lytic resistance of fibrin
Authors: Varjú Imre , Farkas Veronika Judit , Kőhidai László , Szabó László , Farkas Ádám Zoltán , Polgár Lívia , Chinopoulos Christos , Kolev Krasimir
Published: March 2018

In the course of thrombosis, platelets are exposed to a variety of activating stimuli classified as ‘strong’ (e.g. thrombin and collagen) or ‘mild’ (e.g. ADP). In response, activated platelets adhere to injured vasculature, aggregate, and stabilise the three-dimensional fibrin scaffold of the expanding thrombus. Since ‘strong’ stimuli also induce opening of the mitochondrial permeability transition pore (MPTP) in platelets, the MPTP-enhancer Cyclophilin D (CypD) has been suggested as a critical pharmacological target to influence thrombosis. However, it is poorly understood what role CypD plays in the platelet response to ‘mild’ stimuli which act independently of MPTP. Furthermore, it is unknown how CypD influences platelet-driven clot stabilisation against enzymatic breakdown (fibrinolysis). Here we show that treatment of human platelets with Cyclosporine A (a cyclophilin-inhibitor) boosts ADP-induced adhesion and aggregation, while genetic ablation of CypD in murine platelets enhances adhesion but not aggregation. We also report that platelets lacking CypD preserve their integrity in a fibrin environment, and lose their ability to render clots resistant against fibrinolysis. Our results indicate that CypD has opposing haemostatic roles depending on the stimulus and stage of platelet activation, warranting a careful design of any antithrombotic strategy targeting CypD.
Effects of smoking habit change on all-cause mortality and cardiovascular diseases among patients with newly diagnosed diabetes in Korea
Authors: Cho Mi Hee , Lee Kiheon , Park Sang Min , Chang Jooyoung , Choi Seulggie , Kim Kyuwoong , Koo Hye-Yeon , Jun Ji-Hye , Kim Sung Min
Published: March 2018

This study aimed to investigate the effects of smoking habit change on the risks of all-cause mortality and cardiovascular diseases (CVDs) among patients with newly diagnosed diabetes using the Korean National Sample Cohort data. Survival regression analyses for the risks of all-cause mortality and CVDs were performed. Quitters without body mass index (BMI) change (adjusted hazard ratio [aHR], 0.68; 95% confidence interval [CI], 0.46–1.00) and quitters with BMI loss (aHR, 1.76; 95% CI, 1.13–2.73) showed significantly reduced and substantially the increased risk of all-cause mortality, respectively, compared with sustained smokers. Smoking reduction after diabetes diagnosis may have potential positive effects. However, definite benefits on the health outcomes were not identified in this study. Participants who started smoking after diabetes diagnosis had higher risks of all-cause mortality and CVDs than those who were never smokers or ex-smokers, although not statistically significant. In conclusion, smoking cessation after diabetes diagnosis could reduce the risks of all-cause mortality and cardiovascular events among patients with newly diagnosed diabetes when accompanied by proper weight management. Therefore, physicians should advice patients with newly diagnosed type 2 diabetes on the importance of smoking cessation in combination with long-term weight management to maximize the benefits of smoking cessation.
A new artificial diet for western corn rootworm larvae is compatible with and detects resistance to all current Bt toxins
Authors: Ludwick Dalton C. , Meihls Lisa N. , Huynh Man P. , Pereira Adriano E. , French B. Wade , Coudron Thomas A. , Hibbard Bruce E.
Published: March 2018

Insect resistance to transgenic crops is a growing concern for farmers, regulatory agencies, the seed industry, and researchers. Since 2009, instances of field-evolved Bt resistance or cross resistance have been documented for each of the four Bt proteins available for western corn rootworm (WCR), a major insect pest. To characterize resistance, WCR populations causing unexpected damage to Bt maize are evaluated in plant and/or diet toxicity assays. Currently, it is not possible to make direct comparisons of data from different Bt proteins due to differing proprietary artificial diets. Our group has developed a new, publicly available diet (WCRMO-1) with improved nutrition for WCR larvae. For the current manuscript, we tested the compatibility of all Bt proteins currently marketed for WCR on the WCRMO-1 diet and specific proprietary diets corresponding to each toxin using a susceptible colony of WCR. We also tested WCR colonies selected for resistance to each protein to assess the ability of the diet toxicity assay to detect Bt resistance. The WCRMO-1 diet is compatible with each of the proteins and can differentiate resistant colonies from susceptible colonies for each protein. Our diet allows researchers to monitor resistance without the confounding nutritional differences present between diets.
Environmental DNA (eDNA) detects the pool frog (Pelophylax lessonae) at times when traditional monitoring methods are insensitive
Authors: Eiler Alexander , Löfgren Anders , Hjerne Olle , Nordén Sara , Saetre Peter
Published: March 2018

Detection of endangered species is invaluable for conservation efforts, yet many traditional sampling techniques are ineffective at low population abundances or during certain periods of the year. Here, we compared results from a newly developed eDNA approach and the traditional observational method for the endangered pool frog (Pelophylax lessonae). Analysis using an occupancy-modeling framework indicated that the probability of pools being occupied using eDNA (0.93) was higher than for the traditional method of counting calling males and silent observed individuals (0.72). Detailed analysis revealed complementarity among the methods. That is, the traditional method gave a high rate of observation in June, whereas eDNA gave at least as many or more observations during other parts of the year. Discrepancies among the methods depended on the dominant lifecycle stage, and eDNA concentrations were higher when juveniles were present than at times when spawning occurred. eDNA concentrations were also positively related to P. lessonae observations. Our study demonstrates that an eDNA protocol for monitoring of endangered amphibian species can be particularly valuable during periods when individuals are hard to detect by observational methods, and provides guidance to sampling efforts for research and monitoring programs in other regions and systems.
Developing transmission line equations of oxygen transport for predicting oxygen distribution in the arterial system
Authors: Yan Fei , Jiang Wen-Tao , Xu Zhi , Wang Qing-Yuan , Fan Yu-Bo , Zhang Ming
Published: March 2018

The oxygen content in the arterial system plays a significant role in determining the physiological status of a human body. Understanding the oxygen concentration distribution in the arterial system is beneficial for the prevention and intervention of vascular disease. However, the oxygen concentration in the arteries could not be noninvasively monitored in clinical research. Although the oxygen concentration distribution in a vessel could be obtained from a three-dimensional (3D) numerical simulation of blood flow coupled with oxygen transport, a 3D numerical simulation of the systemic arterial tree is complicated and requires considerable computational resources and time. However, the lumped parameter model of oxygen transport derived from transmission line equations of oxygen transport requires fewer computational resources and less time to numerically predict the oxygen concentration distribution in the systemic arterial tree. In this study, transmission line equations of oxygen transport are developed according to the theory of oxygen transport in the vessel, and fluid transmission line equations are used as the theoretical reference for the development. The transmission line equations of oxygen transport could also be regarded as the theoretical basis for developing lumped parameter models of other substances in blood.
Water-Soluble Metalated Covalent Organic Nanobelts with Improved Bioavailability for Protein Transportation
Authors: Kong Weifu , Wan Jiaxun , Namuangruk Supawadee , Guo Jia , Wang Changchun
Published: March 2018

An available pathway to prepare the ionized covalent organic nanosheets (iCONs) has been proposed by a metal-assisted aqueous-phase exfoliation route from covalent organic frameworks. The soluble and belt-shaped iCONs could immobilize a large quantity of proteins (2.73 mg/mg, BSA/iCONs) and hence serve as transporters to enhance the protein uptake by cancer cells. Meanwhile, their energy-dependent endocytosis pathway via clathrin-coated pits has been proved as well.
Involvement of RSK1 activation in malformin-enhanced cellular fibrinolytic activity
Authors: Koizumi Yukio , Nagai Kenichiro , Gao Lina , Koyota Souichi , Yamaguchi Tomokazu , Natsui Miyuki , Imai Yumiko , Hasumi Keiji , Sugiyama Toshihiro , Kuba Keiji
Published: March 2018

Pharmacological interventions to enhance fibrinolysis are effective for treating thrombotic disorders. Utilizing the in vitro U937 cell line-based fibrin degradation assay, we had previously found a cyclic pentapeptide malformin A1 (MA1) as a novel activating compound for cellular fibrinolytic activity. The mechanism by which MA1 enhances cellular fibrinolytic activity remains unknown. In the present study, we show that RSK1 is a crucial mediator of MA1-induced cellular fibrinolysis. Treatment with rhodamine-conjugated MA1 showed that MA1 localizes mainly in the cytoplasm of U937 cells. Screening with an antibody macroarray revealed that MA1 induces the phosphorylation of RSK1 at Ser380 in U937 cells. SL0101, an inhibitor of RSK, inhibited MA1-induced fibrinolytic activity, and CRISPR/Cas9-mediated knockout of RSK1 but not RSK2 suppressed MA1-enhanced fibrinolysis in U937 cells. Synthetic active MA1 derivatives also induced the phosphorylation of RSK1. Furthermore, MA1 treatment stimulated phosphorylation of ERK1/2 and MEK1/2. PD98059, an inhibitor of MEK1/2, inhibited MA1-induced phosphorylation of RSK1 and ERK1/2, indicating that MA1 induces the activation of the MEK-ERK-RSK pathway. Moreover, MA1 upregulated the expression of urokinase-type plasminogen activator (uPA) and increased uPA secretion. These inductions were abrogated in RSK1 knockout cells. These results indicate that RSK1 is a key regulator of MA1-induced extracellular fibrinolytic activity.
Centrifugation-free extraction of circulating nucleic acids using immiscible liquid under vacuum pressure
Authors: Lee Hoyoon , Na Wonhwi , Park Chanhee , Park Kyong Hwa , Shin Sehyun
Published: March 2018

Extraction of cell-free DNA (cfDNA), which exists at an extremely low concentration in plasma, is a critical process for either targeted-sensing or massive sequencing of DNAs. However, such small amount of DNA cannot be fully obtained without high-speed centrifugation (<20,000 g). Here, we developed a centrifugation-free cfDNA extraction method and system that utilizes an immiscible solvent under single low vacuum pressure throughout the entire process. It has been named Pressure and Immiscibility-Based EXtraction (PIBEX). The amounts of extracted cfDNA by PIBEX were compared with those extracted by the conventional gold standards such as QIAGEN using quantitative PCR (qPCR). The PIBEX system showed equal performance regarding extraction amount and efficiency compared to the existing method. Because the PIBEX eliminates the troublous and repetitive centrifugation processes in DNA extraction, it can be further utilized in microfluidic-sample preparation systems for circulating nucleic acids, which would lead to an integrated sample-to-answer system in liquid biopsies.
Source reconstruction of airborne toxics based on acute health effects information
Authors: Argyropoulos Christos D. , Elkhalifa Samar , Fthenou Eleni , Efthimiou George C. , Andronopoulos Spyros , Venetsanos Alexandros , Kovalets Ivan V. , Kakosimos Konstantinos E.
Published: March 2018

The intentional or accidental release of airborne toxics poses great risk to the public health. During these incidents, the greatest factor of uncertainty is related to the location and rate of released substance, therefore, an information of high importance for emergency preparedness and response plans. A novel computational algorithm is proposed to estimate, efficiently, the location and release rate of an airborne toxic substance source based on health effects observations; data that can be readily available, in a real accident, contrary to actual measurements. The algorithm is demonstrated by deploying a semi-empirical dispersion model and Monte Carlo sampling on a simplified scenario. Input data are collected at varying receptor points for toxics concentrations (C; standard approach) and two new types: toxic load (TL) and health effects (HE; four levels). Estimated source characteristics are compared with scenario values. The use ofTLrequired the least number of receptor points to estimate the release rate, and demonstrated the highest probability (>90%).HErequired more receptor points, thanC, but with lesser deviations while probability was comparable, if not better. Finally, the algorithm assessed very accurately the source location when usingCandTLwith comparable confidence, butHEdemonstrated significantly lower confidence.
Involvement of AMP-activated protein kinase and Death Receptor 5 in TRAIL-Berberine-induced apoptosis of cancer cells
Authors: Ke Rong , Vishnoi Kanchan , Viswakarma Navin , Santha Sreevidya , Das Subhasis , Rana Ajay , Rana Basabi
Published: March 2018

Our previous studies indicated that combination of Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and PPARγ ligand Troglitazone (TZD), can induce significant apoptosis in various TRAIL-resistant prostate and hepatocellular carcinoma (HCC) cells. These also suggested serine/threonine kinase AMP-activated protein kinase (AMPK) to be a mediator of TRAIL-TZD-induced apoptosis. To further validate AMPK’s role in TRAIL sensitization, we determined the apoptotic potential of TRAIL in combination with the natural compound Berberine (BBR), the latter being a potent activator of AMPK. These demonstrated a significant reduction of cell viability and induction of apoptosis (increased cleavage of caspase 3, 8, 9) when treated with TRAIL-BBR combination. This apoptosis is attenuated in cells overexpressing AMPKα-dominant negative (DN) or following AMPKα knockdown, confirming involvement of AMPK. To identify potential downstream mediators involved, an apoptosis RT2 PCR array analysis was performed. These showed induction of several genes including TNFRSF10B (expresses DR5) and Harakiri following BBR treatment, which were further validated by qPCR analysis. Furthermore, knocking down DR5 expression significantly attenuated TRAIL-BBR-induced apoptosis, suggesting DR5 to be a mediator of this apoptosis. Our studies indicate that combination of TRAIL and AMPK activator BBR might be an effective means of ameliorating TRAIL-resistance involving DR5 in advanced cancer.
Enhanced Ti0.84Ta0.16N diffusion barriers, grown by a hybrid sputtering technique with no substrate heating, between Si(001) wafers and Cu overlayers
Authors: Mühlbacher Marlene , Greczynski Grzegorz , Sartory Bernhard , Schalk Nina , Lu Jun , Petrov Ivan , Greene J. E. , Hultman Lars , Mitterer Christian
Published: March 2018

We compare the performance of conventional DC magnetron sputter-deposited (DCMS) TiN diffusion barriers between Cu overlayers and Si(001) substrates with Ti0.84Ta0.16N barriers grown by hybrid DCMS/high-power impulse magnetron sputtering (HiPIMS) with substrate bias synchronized to the metal-rich portion of each pulse. DCMS power is applied to a Ti target, and HiPIMS applied to Ta. No external substrate heating is used in either the DCMS or hybrid DCMS/HiPIMS process in order to meet future industrial thermal-budget requirements. Barrier efficiency in inhibiting Cu diffusion into Si(001) while annealing for 1 hour at temperatures between 700 and 900 °C is investigated using scanning electron microscopy, X-ray diffraction, four-point-probe sheet resistance measurements, transmission electron microscopy, and energy-dispersive X-ray spectroscopy profiling. Ti0.84Ta0.16N barriers are shown to prevent large-scale Cu diffusion at temperatures up to 900 °C, while conventional TiN barriers fail at ≤700 °C. The improved performance of the Ti0.84Ta0.16N barrier is due to film densification resulting from HiPIMS pulsed irradiation of the growing film with synchronized Ta ions. The heavy ion bombardment dynamically enhances near-surface atomic mixing during barrier-layer deposition.
Supervised Learning Classifiers for Electrical Impedance-based Bladder State Detection
Authors: Dunne Eoghan , Santorelli Adam , McGinley Brian , Leader Geraldine , O’Halloran Martin , Porter Emily
Published: March 2018

Urinary Incontinence affects over 200 million people worldwide, severely impacting the quality of life of individuals. Bladder state detection technology has the potential to improve the lives of people with urinary incontinence by alerting the user before voiding occurs. To this end, the objective of this study is to investigate the feasibility of using supervised machine learning classifiers to determine the bladder state of ‘full’ or ‘not full’ from electrical impedance measurements. Electrical impedance data was obtained from computational models and a realistic experimental pelvic phantom. Multiple datasets with increasing complexity were formed for varying noise levels in simulation. 10-Fold testing was performed on each dataset to classify ‘full’ and ‘not full’ bladder states, including phantom measurement data. Support vector machines and k-Nearest-Neighbours classifiers were compared in terms of accuracy, sensitivity, and specificity. The minimum and maximum accuracies across all datasets were 73.16% and 100%, respectively. Factors that contributed most to misclassification were the noise level and bladder volumes near the threshold of ‘full’ or ‘not full’. This paper represents the first study to use machine learning for bladder state detection with electrical impedance measurements. The results show promise for impedance-based bladder state detection to support those living with urinary incontinence.
Temperature induced crossing in the optical bandgap of mono and bilayer MoS2 on SiO2
Authors: Park Youngsin , Chan Christopher C. S. , Taylor Robert A. , Kim Yongchul , Kim Nammee , Jo Yongcheol , Lee Seung W. , Yang Woochul , Im Hyunsik , Lee Geunsik
Published: March 2018

Photoluminescence measurements in mono- and bilayer-MoS2 on SiO2 were undertaken to determine the thermal effect of the MoS2/SiO2 interface on the optical bandgap. The energy and intensity of the photoluminescence from monolayer MoS2 were lower and weaker than those from bilayer MoS2 at low temperatures, whilst the opposite was true at high temperatures above 200 K. Density functional theory calculations suggest that the observed optical bandgap crossover is caused by a weaker substrate coupling to the bilayer than to the monolayer.
Foxn1 expression in keratinocytes is stimulated by hypoxia: further evidence of its role in skin wound healing
Authors: Kur-Piotrowska Anna , Bukowska Joanna , Kopcewicz Marta M. , Dietrich Mariola , Nynca Joanna , Slowinska Mariola , Gawronska-Kozak Barbara
Published: March 2018

Recent studies have shown that the transcription factor Foxn1, which is expressed in keratinocytes, is involved in the skin wound healing process, yet how Foxn1 functions remains largely unknown. Our latest data indicate that Foxn1 drives skin healing via engagement in re-epithelization and the epithelial-mesenchymal transition (EMT) process. In the present study, 2D-DIGE proteomic profiling analysis of in vitro cultured keratinocytes transfected with adenoviral vector carrying Foxn1-GFP or GFP alone (control) revealed forty proteins with differential abundance between the compared groups. Among the proteins with Foxn1-dependent expression, several enable adaptation to hypoxia. Subsequent experiments revealed that hypoxic conditions (1% O2) stimulate endogenous and exogenous (transfected Ad-Foxn1) Foxn1 expression in cultured keratinocytes. A proteomics analysis also identified proteins that can act as a factors controlling the balance between cell proliferation, differentiation and apoptosis in response to Foxn1. We also showed that in C57BL/6 keratinocytes, the stimulation of Foxn1 by hypoxia is accompanied by increases in Mmp-9 expression. These data corroborate the detected co-localization of Foxn1 and Mmp-9 expression in vivo in post-wounding skin samples of Foxn1::Egfp transgenic mice. Together, our data indicate that Foxn1 orchestrates cellular changes in keratinocytes in both physiological (self-renewal) and pathological (skin wound healing) contexts.
Epigallocatechin-3-gallate alleviates bladder overactivity in a rat model with metabolic syndrome and ovarian hormone deficiency through mitochondria apoptosis pathways
Authors: Lee Yi-Lun , Lin Kun-Ling , Wu Bin-Nan , Chuang Shu-Mien , Wu Wen-Jeng , Lee Yung-Chin , Ho Wan-Ting , Juan Yung-Shun
Published: March 2018

Metabolic syndrome (MetS) and ovarian hormone deficiency could affect bladder storage dysfunction. Epigallocatechin-3-gallate (EGCG), a polyphenolic compound in green tea, has been shown to protect against ovarian hormone deficiency induced overactive bladder (OAB). The present study investigated oxidative stress induced by MetS and bilateral ovariectomy (OVX), and elucidated the mechanism underlying the protective effect of EGCG (10 umol/kg/day) on bladder overactivity. Rats were fed with high fat high sugar (HFHS) diet to induce MetS and received ovariectomy surgery to deprive ovarian hormone. By dieting with HFHS for 6 months, rats developed MetS and OAB. MetS + OVX deteriorated bladder storage dysfunction more profound than MetS alone. MetS and MetS + OVX rats showed over-expression of inflammatory and fibrosis markers (1.7~3.8-fold of control). EGCG pretreatment alleviated storage dysfunction, and protected the bladders from MetS and OVX - induced interstitial fibrosis changes. Moreover, OVX exacerbated MetS related bladder apoptosis (2.3~4.5-fold of control; 1.8~2.6-fold of Mets group), enhances oxidative stress markers (3.6~4.3-fold of control; 1.8~2.2-fold of Mets group) and mitochondrial enzyme complexes subunits (1.8~3.7-fold of control; 1.5~3.4-fold of Mets group). EGCG pretreatment alleviated bladder apoptosis, attenuated oxidative stress, and reduced the mitochondrial and endoplasmic reticulum apoptotic signals. In conclusions, HFHS feeding and ovarian hormone deficiency enhances the generation of oxidative stress mediated through mitochondrial pathway. EGCG reduced the generation of oxidative stress and lessened bladder overactivity.
Exploring the capabilities of monochromated electron energy loss spectroscopy in the infrared regime
Authors: Hachtel Jordan A. , Lupini Andrew R. , Idrobo Juan Carlos
Published: March 2018

Monochromated electron energy loss spectroscopy (EELS) is one of the leading techniques to study materials properties that correspond to low (<5 eV) energy losses (i.e. band-gaps, plasmons, and excitons) with nanoscale spatial resolution. Recently a new generation of monochromators have become available, opening regimes and unlocking excitations that were previously unobservable in the electron microscope. The capabilities of these new instruments are still being explored, and here we study the effect of monochromation on various aspects of EELS analysis in the infrared (<1 eV) regime. We investigate the effect of varying levels of monochromation on energy resolution, zero-loss peak (ZLP) tail reduction, ZLP tail shape, signal-to-noise-ratio, and spatial resolution. From these experiments, the new capabilities of monochromated EELS are shown to be highly promising for the future of localized spectroscopic analysis.
Rapid selection of sulphadoxine-resistant Plasmodium falciparum and its effect on within-population genetic diversity in Papua New Guinea
Authors: Mita Toshihiro , Hombhanje Francis , Takahashi Nobuyuki , Sekihara Makoto , Yamauchi Masato , Tsukahara Takahiro , Kaneko Akira , Endo Hiroyoshi , Ohashi Jun
Published: March 2018

The ability of the human malarial parasite Plasmodium falciparum to adapt to environmental changes depends considerably on its ability to maintain within-population genetic variation. Strong selection, consequent to widespread antimalarial drug usage, occasionally elicits a rapid expansion of drug-resistant isolates, which can act as founders. To investigate whether this phenomenon induces a loss of within-population genetic variation, we performed a population genetic analysis on 302 P. falciparum cases detected during two cross-sectional surveys in 2002/2003, just after the official introduction of sulphadoxine/pyrimethamine as a first-line treatment, and again in 2010/2011, in highly endemic areas in Papua New Guinea. We found that a single-origin sulphadoxine-resistant parasite isolate rapidly increased from 0% in 2002/2003 to 54% in 2010 and 84% in 2011. However, a considerable number of pairs exhibited random associations among 10 neutral microsatellite markers located in various chromosomes, suggesting that outcrossing effectively reduced non-random associations, albeit at a low average multiplicity of infection (1.35–1.52). Within-population genetic diversity was maintained throughout the study period. This indicates that the parasites maintained within-population variation, even after a clonal expansion of drug-resistant parasites. Outcrossing played a role in the preservation of within-population genetic diversity despite low levels of multiplicity of infection.
In-situ Observation of Hierarchical Self-Assembly Driven by Bicontinuous Gelation in Mixed Nanodisc Dispersions
Authors: Pujala Ravi Kumar , Schneijdenberg C. T. W. M. , van Blaaderen Alfons , Bohidar H. B.
Published: March 2018

The search for new functional soft materials with precise and reconfigurable structures at the nano and meso-scale is a major challenge as well as objective of the current science. Patchy colloids of different shapes and functionalities are considered important new building blocks of a bottom-up approach towards rational design of new soft materials largely governed by anisotropic interactions. Herein, we investigate the self-assembly, growth of hierarchical microstructures and aging dynamics of 2D nano-platelets of two different aspect ratios (Laponite ~25 and Montmorillonite ~250) which form gels with different porosity that is achieved by tuning their mixing ratios. Qualitative in situ real-space studies are carried out, including fluorescent confocal microscopy imaging of the bicontinuous gelation process or final states, which provides dynamic visualization of the self-organization. The bicontinuous gels exhibit a foam-like morphology having pores of a few micrometers in size that can be tuned by varying the mixing ratio of nanoplatelets. It is shown that this new class of clay gels has unique and tunable physical properties that will find potential applications in the development of low cost lithium ion batteries, nanocomposites and nuclear waste management.
Enhanced labeling density and whole-cell 3D dSTORM imaging by repetitive labeling of target proteins
Authors: Venkataramani Varun , Kardorff Markus , Herrmannsdörfer Frank , Wieneke Ralph , Klein Alina , Tampé Robert , Heilemann Mike , Kuner Thomas
Published: March 2018

With continuing advances in the resolving power of super-resolution microscopy, the inefficient labeling of proteins with suitable fluorophores becomes a limiting factor. For example, the low labeling density achieved with antibodies or small molecule tags limits attempts to reveal local protein nano-architecture of cellular compartments. On the other hand, high laser intensities cause photobleaching within and nearby an imaged region, thereby further reducing labeling density and impairing multi-plane whole-cell 3D super-resolution imaging. Here, we show that both labeling density and photobleaching can be addressed by repetitive application of trisNTA-fluorophore conjugates reversibly binding to a histidine-tagged protein by a novel approach called single-epitope repetitive imaging (SERI). For single-plane super-resolution microscopy, we demonstrate that, after multiple rounds of labeling and imaging, the signal density is increased. Using the same approach of repetitive imaging, washing and re-labeling, we demonstrate whole-cell 3D super-resolution imaging compensated for photobleaching above or below the imaging plane. This proof-of-principle study demonstrates that repetitive labeling of histidine-tagged proteins provides a versatile solution to break the ‘labeling barrier’ and to bypass photobleaching in multi-plane, whole-cell 3D experiments.
Effects of methamphetamine abuse on spatial cognitive function
Authors: Luo Yan-Lin , Bian Jing-Wei , Zheng Zhi-Jun , Zhao Li , Han Song , Sun Xiao-Hong , Li Jun-Fa , Ni Guo-Xin
Published: March 2018

Methamphetamine (MA) abuse has been rising rapidly over the past decade, however, its impact in spatial cognitive function remains unknown. To understand its effect on visuospatial ability and spatial orientation ability, 40 MA users and 40 non-MA users conducted the Simple Reaction Task (Task 1), the Spatial Orientation Task (Task 2), and the Mental Rotation Task (Task 3), respectively. There was no significant difference in either accuracy or reaction time (RT) between 2 groups in Task 1. During Task 2, in comparison with non-MA users, MA users performed poorer on RT, but not in accuracy for foot and hand stimuli. In addition, both non-MA and MA users responded much more quickly to upward stimuli than downward stimuli on vertical surface, however, only non-MA users exhibited leftward visual field advantage in horizontal orientation processing. As for Task 3, MA users exhibited poorer performance and more errors than their healthy counterparts. For each group, linear relationship was revealed between RT and orientation angle, whereas MA abuse led to longer intercept for all stimuli involved. Our findings suggested that MA abuse may lead to a general deficit in the visuospatial ability and the spatial orientation ability with more serious impact in the former.
Drug resistance of oral bacteria to new antibacterial dental monomer dimethylaminohexadecyl methacrylate
Authors: Wang Suping , Wang Haohao , Ren Biao , Li Xiaodong , Wang Lin , Zhou Han , Weir Michael D. , Zhou Xuedong , Masri Radi M. , Oates Thomas W. , Cheng Lei , Xu Hockin H. K.
Published: March 2018

Only two reports exist on drug-resistance of quaternary ammonium monomers against oral bacteria; both studies tested planktonic bacteria for 10 passages, and neither study tested biofilms or resins. The objectives of this study were to investigate the drug-resistance of Streptococcus mutans, Streptococcus sanguinis and Streptococcus gordonii against dimethylaminohexadecyl methacrylate (DMAHDM), and to evaluate biofilms on resins with repeated exposures for 20 passages for the first time. DMAHDM, dimethylaminododecyl methacrylate (DMADDM) and chlorhexidine (CHX) were tested with planktonic bacteria. Biofilms were grown on a resin containing 3% DMAHDM. Minimum-inhibitory concentrations were measured. To detect drug-resistance, the survived bacteria from the previous passage were used as inoculum for the next passage for repeated exposures. S. gordonii developed drug-resistance against DMADDM and CHX, but not against DMAHDM. Biofilm colony-forming units (CFU) on DMAHDM-resin was reduced by 3–4 log; there was no difference from passages 1 to 20 (p > 0.1). No drug-resistance to DMAHDM was detected for all three bacterial species. In conclusion, this study showed that DMAHDM induced no drug-resistance, and DMAHDM-resin reduced biofilm CFU by 3–4 log, with no significant change from 1 to 20 passages. DMAHDM with potent antibacterial activities and no drug-resistance is promising for dental applications.
Interrupted breeding in a songbird migrant triggers development of nocturnal locomotor activity
Authors: Mukhin Andrey , Kobylkov Dmitry , Kishkinev Dmitry , Grinkevich Vitaly
Published: March 2018

Long-distance avian migrants, e.g. Eurasian reed warblers (Acrocephalus scirpaceus), can precisely schedule events of their annual cycle. However, the proximate mechanisms controlling annual cycle and their interplay with environmental factors are poorly understood. We artificially interrupted breeding in reed warblers by bringing them into captivity and recording birds’ locomotor activity for 5–7 days. Over this time, most of the captive birds gradually developed nocturnal locomotor activity not observed in breeding birds. When the birds were later released and radio-tracked, the individuals with highly developed caged activity performed nocturnal flights. We also found that reed warblers kept indoors without access to local cues developed a higher level of nocturnal activity compared to the birds kept outdoors with an access to the familiar environment. Also, birds translocated from a distant site (21 km) had a higher motivation to fly at night-time after release compared to the birds captured within 1 km of a study site. Our study suggests that an interrupted breeding triggers development of nocturnal locomotor activity in cages, and the level of activity is correlated with motivation to perform nocturnal flights in the wild, which can be restrained by familiar environment.
Long-term Western diet fed apolipoprotein E-deficient rats exhibit only modest early atherosclerotic characteristics
Authors: Rune Ida , Rolin Bidda , Lykkesfeldt Jens , Nielsen Dennis Sandris , Krych Łukasz , Kanter Jenny E. , Bornfeldt Karin E. , Kihl Pernille , Buschard Karsten , Josefsen Knud , Fels Johannes Josef , Mortensen Alan , Christoffersen Berit , Kirk Rikke Kaae , Hansen Axel Kornerup
Published: March 2018

In the apolipoprotein E–deficient mouse, the gut microbiota has an impact on the development of atherosclerosis, but whether such correlations are also present in rats requires investigation. Therefore, we studied female SD-Apoe
tm1sage
(Apoe−/−) rats fed either a Western diet or a low-fat control diet with or without gluten, which is known to promote gut microbiota changes, until 20 weeks of age. We hypothesized that the manifestation of atherosclerosis would be more severe in Apoe−/− rats fed the Western high-fat diet, as compared with rats fed the low-fat diet, and that atherosclerosis would be accelerated by gluten. Both Western diet-feeding and gluten resulted in significant changes in gut microbiota, but the microbiota impact of gluten was transient. Compared with Apoe−/− rats fed a low-fat diet, Western diet-fed Apoe−/− rats were heavier and became glucose intolerant with increased levels of oxidative stress. They developed early fatty streak lesions in their aortic sinus, while there was no evidence of atherosclerosis in the thoracic aorta. No conclusions could be made on the impact of gluten on atherosclerosis. Although Western diet-fed Apoe−/− rats exhibited a more human-like LDL dominated blood lipid profile, signs of obesity, type 2 diabetes and cardiovascular disease were modest.
Causal Inference in the Perception of Verticality
Authors: de Winkel Ksander N. , Katliar Mikhail , Diers Daniel , Bülthoff Heinrich H.
Published: March 2018

The perceptual upright is thought to be constructed by the central nervous system (CNS) as a vector sum; by combining estimates on the upright provided by the visual system and the body’s inertial sensors with prior knowledge that upright is usually above the head. Recent findings furthermore show that the weighting of the respective sensory signals is proportional to their reliability, consistent with a Bayesian interpretation of a vector sum (Forced Fusion, FF). However, violations of FF have also been reported, suggesting that the CNS may rely on a single sensory system (Cue Capture, CC), or choose to process sensory signals based on inferred signal causality (Causal Inference, CI). We developed a novel alternative-reality system to manipulate visual and physical tilt independently. We tasked participants (n = 36) to indicate the perceived upright for various (in-)congruent combinations of visual-inertial stimuli, and compared models based on their agreement with the data. The results favor the CI model over FF, although this effect became unambiguous only for large discrepancies (±60°). We conclude that the notion of a vector sum does not provide a comprehensive explanation of the perception of the upright, and that CI offers a better alternative.
Combining streptozotocin and unilateral nephrectomy is an effective method for inducing experimental diabetic nephropathy in the ‘resistant’ C57Bl/6J mouse strain
Authors: Uil Melissa , Scantlebery Angelique M. L. , Butter Loes M. , Larsen Per W. B. , de Boer Onno J. , Leemans Jaklien C. , Florquin Sandrine , Roelofs Joris J. T. H.
Published: March 2018

Diabetic nephropathy (DN) is the leading cause of chronic kidney disease. Animal models are essential tools for designing new strategies to prevent DN. C57Bl/6 (B6) mice are widely used for transgenic mouse models, but are relatively resistant to DN. This study aims to identify the most effective method to induce DN in a type 1 (T1D) and a type 2 diabetes (T2D) model in B6 mice. For T1D-induced DN, mice were fed a control diet, and randomised to streptozotocin (STZ) alone, STZ+unilateral nephrectomy (UNx), or vehicle/sham. For T2D-induced DN, mice were fed a western (high fat) diet, and randomised to either STZ alone, STZ+UNx, UNx alone, or vehicle/sham. Mice subjected to a control diet with STZ +UNx developed albuminuria, glomerular lesions, thickening of the glomerular basement membrane, and tubular injury. Mice on control diet and STZ developed only mild renal lesions. Furthermore, kidneys from mice on a western diet were hardly affected by diabetes, UNx or the combination. We conclude that STZ combined with UNx is the most effective model to induce T1D-induced DN in B6 mice. In our hands, combining western diet and STZ treatment with or without UNx did not result in a T2D-induced DN model in B6 mice.
Epigenetic alterations are associated with monocyte immune dysfunctions in HIV-1 infection
Authors: Espíndola Milena S. , Soares Luana S. , Galvão-Lima Leonardo J. , Zambuzi Fabiana A. , Cacemiro Maira C. , Brauer Verônica S. , Marzocchi-Machado Cleni M. , de Souza Gomes Matheus , Amaral Laurence R. , Martins-Filho Olindo A. , Bollela Valdes R. , Frantz Fabiani G.
Published: March 2018

Monocytes are key cells in the immune dysregulation observed during human immunodeficiency virus (HIV) infection. The events that take place specifically in monocytes may contribute to the systemic immune dysfunction characterized by excessive immune activation in infected individuals, which directly correlates with pathogenesis and progression of the disease. Here, we investigated the immune dysfunction in monocytes from untreated and treated HIV + patients and associated these findings with epigenetic changes. Monocytes from HIV patients showed dysfunctional ability of phagocytosis and killing, and exhibited dysregulated cytokines and reactive oxygen species production afterM. tuberculosischallengein vitro. In addition, we showed that the expression of enzymes responsible for epigenetic changes was altered during HIV infection and was more prominent in patients that had high levels of soluble CD163 (sCD163), a newly identified plasmatic HIV progression biomarker. Among the enzymes, histone acetyltransferase 1 (HAT1) was the best epigenetic biomarker correlated with HIV - sCD163 high patients. In conclusion, we confirmed that HIV impairs effector functions of monocytes and these alterations are associated with epigenetic changes that once identified could be used as targets in therapies aiming the reduction of the systemic activation state found in HIV patients.
Retrospective Study of Hemodynamic Changes Before and After Carotid Stenosis Formation by Vessel Surface Repairing
Authors: Li Xiao , Sun Beibei , Zhao Huilin , Ge Xiaoqian , Liang Fuyou , Li Xuanyu , Xu Jianrong , Liu Xiaosheng
Published: March 2018

Prospective observation of hemodynamic changes before and after the formation of atherosclerotic stenosis in the carotid artery is difficult. Thus, a vessel surface repairing method was used for retrospective hemodynamic study before and after atherosclerotic stenosis formation in carotid artery. The three-dimensional geometry of sixteen sinus atherosclerotic stenosis carotid arteries were repaired and restored as normal arteries. Computational fluid dynamics analysis was performed to estimate wall shear stress (WSS), velocity and vortex in atherosclerosis-free areas and sinus in stenosis-repaired carotid artery. The analysis was also performed in the stenotic segment and upstream and downstream of stenosis in stenotic carotid artery. Compared to the atherosclerosis-free areas in stenosis-repaired carotid artery, sinus presented significantly lower WSS (P < 0.05), lower velocity (P < 0.05) and apparent vortex. Compared to the sinus, the WSS in the upstream of stenosis was lower (P < 0.05), while in the downstream area was similar (P = 0.87), both upstream and downstream of stenosis demonstrated similar velocity to sinus (P = 0.76 and P = 0.36, respectively) and apparent vortex. Atherosclerosis-prone areas including normal carotid sinus and upstream and downstream of stenosis in stenotic carotid artery were subjected to lower WSS and velocity as well as apparent vortex, thereby might be associated with the formation and progress of atherosclerosis.
Discrimination between the human prostate normal and cancer cell exometabolome by GC-MS
Authors: Lima Ana Rita , Araújo Ana Margarida , Pinto Joana , Jerónimo Carmen , Henrique Rui , Bastos Maria de Lourdes , Carvalho Márcia , Guedes de Pinho Paula
Published: March 2018

Serum prostate-specific antigen (PSA) is currently the most used biomarker in clinical practice for prostate cancer (PCa) detection. However, this biomarker has several drawbacks. In this work, an untargeted gas chromatography-mass spectrometry (GC-MS)-based metabolomic profiling of PCa cells was performed to prove the concept that metabolic alterations might differentiate PCa cell lines from normal prostate cell line. For that, we assessed the differences in volatile organic compounds (VOCs) profile in the extracellular medium (exometabolome) of four PCa cell lines and one normal prostate cell line at two pH values (pH 2 and 7) by GC-MS. Multivariate analysis revealed a panel of volatile metabolites that discriminated cancerous from normal prostate cells. The most altered metabolites included ketones, aldehydes and organic acids. Among these, we highlight pentadecane-2-one and decanoic acid, which were significantly increased in PCa compared to normal cells, and cyclohexanone, 4-methylheptan-2-one, 2-methylpentane-1,3-diol, 4-methylbenzaldehyde, 1-(3,5-dimethylfuran-2-yl)ethanone, methyl benzoate and nonanoic acid, which were significantly decreased in PCa cells. The PCa volatilome was markedly influenced by the VOCs extraction pH, though the discriminant capability was similar. Overall, our data suggest that VOCs monitoring has the potential to be used as a PCa screening methodology.
Activation of the complement system in an osteosarcoma cell line promotes angiogenesis through enhanced production of growth factors
Authors: Jeon Hyungtaek , Han Seung Ro , Lee Suhyuk , Park Sang June , Kim Joo Heon , Yoo Seung-Min , Lee Myung-Shin
Published: March 2018

There is increasing evidence that the complement system is activated in various cancer tissues. Besides being involved in innate immunity against pathogens, the complement system also participates in inflammation and the modulation of tumor microenvironment. Recent studies suggest that complement activation promotes tumor progression in various ways. Among some cancer cell lines, we found that human bone osteosarcoma epithelial cells (U2-OS) can activate the alternative pathway of the complement system by pooled normal human serum. Interestingly, U2-OS cells showed less expression of complement regulatory proteins, compared to other cancer cell lines. Furthermore, the activated complement system enhanced the production of growth factors, which promoted angiogenesis of human endothelial cells. Our results demonstrated a direct linkage between the complement system and angiogenesis using the in vitro model, which suggest the complement system and related mechanisms might be potential targets for cancer treatment.
An intronic mutation in Chd7 creates a cryptic splice site, causing aberrant splicing in a mouse model of CHARGE syndrome
Authors: Ogier Jacqueline M. , Arhatari Benedicta D. , Carpinelli Marina R. , McColl Bradley K. , Wilson Michael A. , Burt Rachel A.
Published: March 2018

Alternate splicing is a critical regulator of gene expression in eukaryotes, however genetic mutations can cause erroneous splicing and disease. Most recorded splicing disorders are caused by mutations of splice donor/acceptor sites, however intronic mutations can affect splicing. Clinical exome analyses largely ignore intronic sequence, limiting the detection of mutations to within coding regions. We describe ‘Trooper’, a novel mouse model of CHARGE syndrome harbouring a pathogenic point mutation in Chd7. The mutation is 18 nucleotides upstream of exon 10 and creates a cryptic acceptor site, causing exon skipping and partial intron retention. This mutation, though detectable in exome sequence, was initially dismissed by computational filtering due to its intronic location. The Trooper strain exhibited many of the previously described CHARGE-like anomalies of CHD7 deficient mouse lines; including hearing impairment, vestibular hypoplasia and growth retardation. However, more common features such as facial asymmetry and circling were rarely observed. Recognition of these characteristic features prompted manual reexamination of Chd7 sequence and subsequent validation of the intronic mutation, highlighting the importance of phenotyping alongside exome analyses. The Trooper mouse serves as a valuable model of atypical CHARGE syndrome and reveals a molecular mechanism that may underpin milder clinical presentation of the syndrome.