bluuebunny/crossref_metadata_2025 huggingface dataset:
Caspase-1-dependent inflammasomes mediate photoreceptor cell death in photo-oxidative damage-induced retinal degeneration
Authors: Wooff Yvette , Fernando Nilisha , Wong Josephine H. C. , Dietrich Catherine , Aggio-Bruce Riemke , Chu-Tan Joshua A. , Robertson Avril A. B. , Doyle Sarah L. , Man Si Ming , Natoli Riccardo
Published: February 2020

Activation of the inflammasome is involved in the progression of retinal degenerative diseases, in particular, in the pathogenesis of Age-Related Macular Degeneration (AMD), with NLRP3 activation the focus of many investigations. In this study, we used genetic and pharmacological approaches to explore the role of the inflammasome in a mouse model of retinal degeneration. We identify that Casp1/11−/− mice have better-preserved retinal function, reduced inflammation and increased photoreceptor survivability. While Nlrp3−/− mice display some level of preservation of retinal function compared to controls, pharmacological inhibition of NLRP3 did not protect against photoreceptor cell death. Further, Aim2−/−, Nlrc4−/−, Asc−/−, and Casp11−/− mice show no substantial retinal protection. We propose that CASP-1-associated photoreceptor cell death occurs largely independently of NLRP3 and other established inflammasome sensor proteins, or that inhibition of a single sensor is not sufficient to repress the inflammatory cascade. Therapeutic targeting of CASP-1 may offer a more promising avenue to delay the progression of retinal degenerations.
Wild blueberry proanthocyanidins shape distinct gut microbiota profile and influence glucose homeostasis and intestinal phenotypes in high-fat high-sucrose fed mice
Authors: Rodríguez-Daza Maria-Carolina , Daoust Laurence , Boutkrabt Lemia , Pilon Geneviève , Varin Thibault , Dudonné Stéphanie , Levy Émile , Marette André , Roy Denis , Desjardins Yves
Published: February 2020

Blueberries are a rich source of polyphenols, widely studied for the prevention or attenuation of metabolic diseases. However, the health contribution and mechanisms of action of polyphenols depend on their type and structure. Here, we evaluated the effects of a wild blueberry polyphenolic extract (WBE) (Vaccinium angustifolium Aiton) on cardiometabolic parameters, gut microbiota composition and gut epithelium histology of high-fat high-sucrose (HFHS) diet-induced obese mice and determined which constitutive polyphenolic fractions (BPF) was responsible for the observed effects. To do so, the whole extract was separated in three fractions, F1) Anthocyanins and phenolic acids, F2) oligomeric proanthocyanidins (PACs), phenolic acids and flavonols (PACs degree of polymerization DP < 4), and F3) PACs polymers (PACs DP > 4) and supplied at their respective concentration in the whole extract. After 8 weeks, WBE reduced OGTT AUC by 18.3% compared to the HFHS treated rodents and the F3 fraction  contributed the most to this effect. The anthocyanin rich F1 fraction did not reproduce this response. WBE and the BPF restored the colonic mucus layer. Particularly, the polymeric PACs-rich F3 fraction increased the mucin-secreting goblet cells number. WBE caused a significant 2-fold higher proportion of Adlercreutzia equolifaciens whereas oligomeric PACs-rich F2 fraction increased by 2.5-fold the proportion of Akkermansia muciniphila. This study reveals the key role of WBE PACs in modulating the gut microbiota and restoring colonic epithelial mucus layer, providing a suitable ecological niche for mucosa-associated symbiotic bacteria, which may be crucial in triggering health effects of blueberry polyphenols.
Nandrolone induces a stem cell-like phenotype in human hepatocarcinoma-derived cell line inhibiting mitochondrial respiratory activity
Authors: Agriesti Francesca , Tataranni Tiziana , Pacelli Consiglia , Scrima Rosella , Laurenzana Ilaria , Ruggieri Vitalba , Cela Olga , Mazzoccoli Carmela , Salerno Monica , Sessa Francesco , Sani Gabriele , Pomara Cristoforo , Capitanio Nazzareno , Piccoli Claudia
Published: February 2020

Nandrolone is a testosterone analogue with anabolic properties commonly abused worldwide, recently utilized also as therapeutic agent in chronic diseases, cancer included. Here we investigated the impact of nandrolone on the metabolic phenotype in HepG2 cell line. The results attained show that pharmacological dosage of nandrolone, slowing cell growth, repressed mitochondrial respiration, inhibited the respiratory chain complexes I and III and enhanced mitochondrial reactive oxygen species (ROS) production. Intriguingly, nandrolone caused a significant increase of stemness-markers in both 2D and 3D cultures, which resulted to be CxIII-ROS dependent. Notably, nandrolone negatively affected differentiation both in healthy hematopoietic and mesenchymal stem cells. Finally, nandrolone administration in mice confirmed the up-regulation of stemness-markers in liver, spleen and kidney. Our observations show, for the first time, that chronic administration of nandrolone, favoring maintenance of stem cells in different tissues would represent a precondition that, in addition to multiple hits, might enhance risk of carcinogenesis raising warnings about its abuse and therapeutic utilization.
PHTNet: Characterization and Deep Mining of Involuntary Pathological Hand Tremor using Recurrent Neural Network Models
Authors: Shahtalebi Soroosh , Atashzar Seyed Farokh , Samotus Olivia , Patel Rajni V. , Jog Mandar S. , Mohammadi Arash
Published: February 2020

The global aging phenomenon has increased the number of individuals with age-related neurological movement disorders including Parkinson’s Disease (PD) and Essential Tremor (ET). Pathological Hand Tremor (PHT), which is considered among the most common motor symptoms of such disorders, can severely affect patients’ independence and quality of life. To develop advanced rehabilitation and assistive technologies, accurate estimation/prediction of nonstationary PHT is critical, however, the required level of accuracy has not yet been achieved. The lack of sizable datasets and generalizable modeling techniques that can fully represent the spectrotemporal characteristics of PHT have been a critical bottleneck in attaining this goal. This paper addresses this unmet need through establishing a deep recurrent model to predict and eliminate the PHT component of hand motion. More specifically, we propose a machine learning-based, assumption-free, and real-time PHT elimination framework, the PHTNet, by incorporating deep bidirectional recurrent neural networks. The PHTNet is developed over a hand motion dataset of 81 ET and PD patients collected systematically in a movement disorders clinic over 3 years. The PHTNet is the first intelligent systems model developed on this scale for PHT elimination that maximizes the resolution of estimation and allows for prediction of future and upcoming sub-movements.
An optimized workflow for single-cell transcriptomics and repertoire profiling of purified lymphocytes from clinical samples
Authors: Hanamsagar Richa , Reizis Timothy , Chamberlain Mathew , Marcus Robert , Nestle Frank O. , de Rinaldis Emanuele , Savova Virginia
Published: February 2020

Establishing clinically relevant single-cell (SC) transcriptomic workflows from cryopreserved tissue is essential to move this emerging immune monitoring technology from the bench to the bedside. Improper sample preparation leads to detrimental cascades, resulting in loss of precious time, money and finally compromised data. There is an urgent need to establish protocols specifically designed to overcome the inevitable variations in sample quality resulting from uncontrollable factors in a clinical setting. Here, we explore sample preparation techniques relevant to a range of clinically relevant scenarios, where SC gene expression and repertoire analysis are applied to a cryopreserved sample derived from a small amount of blood, with unknown or partially known preservation history. We compare a total of ten cell-counting, viability-improvement, and lymphocyte-enrichment methods to highlight a number of unexpected findings. Trypan blue-based automated counters, typically recommended for single-cell sample quantitation, consistently overestimate viability. Advanced sample clean-up procedures significantly impact total cell yield, while only modestly increasing viability. Finally, while pre-enrichment of B cells from whole peripheral blood mononuclear cells (PBMCs) results in the most reliable BCR repertoire data, comparable T-cell enrichment strategies distort the ratio of CD4+ and CD8+ cells. Furthermore, we provide high-resolution analysis of gene expression and clonotype repertoire of different B cell subtypes. Together these observations provide both qualitative and quantitative sample preparation guidelines that increase the chances of obtaining high-quality single-cell transcriptomic and repertoire data from human PBMCs in a variety of clinical settings.
Correlation between adenoma detection rate and polyp detection rate at endoscopy in a non-screening population
Authors: Murphy B. , Myers E. , O’Shea T. , Feeley K. , Waldron B.
Published: February 2020

It is understood that colorectal adenomas progress to colonic adenocarcinoma. Adenoma detection rate (ADR) at endoscopy has been used as a key performance indicator at endoscopy and is inversely associated with diagnosis of interval colorectal cancer. As most endoscopy reporting systems do not routinely incorporate histological assessment, ADR reporting is a cumbersome task. Polyp Detection Rate (PDR) has therefore been adopted as a surrogate marker for ADR. A prospectively maintained database of colonoscopies performed between July 2015 and July 2017 was analysed. This was cross referenced with a histological database. Statistical analysis was performed using IBM SPSS, version 24. Inferential procedures employed included the Pearson’s correlation coefficient (r) and Binomial logistic regression. Of 2964 procedures performed by 8 endoscopists, overall PDR was 27% and ADR was 19%. The PDR, ADR, adenoma to polyp detection rate quotient (APDRQ) and estimated ADR (PDR x APDRQ group average = 0.72) was calculated for each individual. There was a strong positive linear correlation between PDR and ADR,r(8) = 0.734, p = 0.038 and between PDR and estimated ADR, r(8) = 0.998, p < 0.001. Adenoma detection rate strongly correlated with estimated ADR, r(8) = 0.720, p = 0.044. With the exclusion of a moderate outlier, these correlations increased in both strength and significance. There was a stronger correlation between PDR and ADR,r(7) = 0.921, p = 0.003 and between ADR and estimated ADR, r(7) = 0.928, p = 0.003.
A monolithic nanostructured-perovskite/silicon tandem solar cell: feasibility of light management through geometry and materials selection
Authors: Elshorbagy Mahmoud H. , López-Fraguas Eduardo , Chaudhry Fateh A. , Sánchez-Pena José Manuel , Vergaz Ricardo , García-Cámara Braulio
Published: February 2020

The use of several layers of different materials, taking advantage of their complementary bandgap energies, improves the absorption in multi-junction solar cells. Unfortunately, the inherent efficiency increment of this strategy has a limitation: each interface introduces optical losses. In this paper, we study the effects of materials and geometry in the optical performance of a nanostructured hybrid perovskite – silicon tandem solar cell. Our proposed design increases the performance of both subcells by managing light towards the active layer, as well as by minimizing reflections losses in the interfaces. We sweep both refractive index and thickness of the transport layers and the dielectric spacer composing the metasurface, obtaining a range of these parameters for the proper operation of the device. Using these values, we obtain a reduction in the optical losses, in particular they are more than a 33% lower than those of a planar cell, mainly due to a reduction of the reflectivity in the device. This approach leads to an enhancement in the optical response, widens the possibilities for the manufacturers to use different materials, and allows wide geometrical tolerances.
Statistical learning for vocal sequence acquisition in a songbird
Authors: James Logan S. , Sun Herie , Wada Kazuhiro , Sakata Jon T.
Published: February 2020

Birdsong is a learned communicative behavior that consists of discrete acoustic elements (“syllables”) that are sequenced in a controlled manner. While the learning of the acoustic structure of syllables has been extensively studied, relatively little is known about sequence learning in songbirds. Statistical learning could contribute to the acquisition of vocal sequences, and we investigated the nature and extent of sequence learning at various levels of song organization in the Bengalese finch, Lonchura striata var. domestica. We found that, under semi-natural conditions, pupils (sons) significantly reproduced the sequence statistics of their tutor’s (father’s) songs at multiple levels of organization (e.g., syllable repertoire, prevalence, and transitions). For example, the probability of syllable transitions at “branch points” (relatively complex sequences that are followed by multiple types of transitions) were significantly correlated between the songs of tutors and pupils. We confirmed the contribution of learning to sequence similarities between fathers and sons by experimentally tutoring juvenile Bengalese finches with the songs of unrelated tutors. We also discovered that the extent and fidelity of sequence similarities between tutors and pupils were significantly predicted by the prevalence of sequences in the tutor’s song and that distinct types of sequence modifications (e.g., syllable additions or deletions) followed distinct patterns. Taken together, these data provide compelling support for the role of statistical learning in vocal production learning and identify factors that could modulate the extent of vocal sequence learning.
Porous Si-SiO2 based UV Microcavities
Authors: Jimenéz-Vivanco María R. , García Godofredo , Carrillo Jesús , Agarwal Vivechana , Díaz-Becerril Tomás , Doti Rafael , Faubert Jocelyn , Lugo J. E.
Published: February 2020

Obtaining silicon-based photonic-structures in the ultraviolet range would expand the wavelength bandwidth of silicon technology, where it is normally forbidden. Herein, we fabricated porous silicon microcavities by electrochemical etching of alternating high and low refraction index layers; and were carefully subjected to two stages of dry oxidation at 350 °C for 30 minutes and 900 °C, with different oxidation times. In this way, we obtained oxidized porous silicon that induces a shift of a localized mode in the ultraviolet region. The presence of Si-O-Si bonds was made clear by FTIR absorbance spectra. High-quality oxidized microcavities were shown by SEM, where their mechanical stability was clearly visible. We used an effective medium model to predict the refractive index and optical properties of the microcavities. The model can use either two or three components (Si, SiO2, and air). The latter predicts that the microcavities are made almost completely of SiO2, implying less photon losses in the structure. The theoretical photonic-bandgap structure and localized photonic mode location showed that the experimental spectral peaks within the UV photonic bandgap are indeed localized modes. These results support that our oxidation process is very advantageous to obtain complex photonic structures in the UV region.
SIAH2-mediated and organ-specific restriction of HO-1 expression by a dual mechanism
Authors: Chillappagari Shashipavan , Belapurkar Ratnal , Möller Andreas , Molenda Nicole , Kracht Michael , Rohrbach Susanne , Schmitz M. Lienhard
Published: February 2020

The intracellular levels of the cytoprotective enzyme heme oxygenase-1 (HO-1) are tightly controlled. Here, we reveal a novel mechanism preventing the exaggerated expression of HO-1. The analysis of mice with a knock-out in the ubiquitin E3 ligase seven in absentia homolog 2 (SIAH2) showed elevated HO-1 protein levels in specific organs such as heart, kidney and skeletal muscle. Increased HO-1 protein amounts were also seen in human cells deleted for the SIAH2 gene. The higher HO-1 levels are not only due to an increased protein stability but also to elevated expression of the HO-1 encoding HMOX1 gene, which depends on the transcription factor nuclear factor E2-related factor 2 (NRF2), a known SIAH2 target. Dependent on its RING (really interesting new gene) domain, expression of SIAH2 mediates proteasome-dependent degradation of its interaction partner HO-1. Additionally SIAH2-deficient cells are also characterized by reduced expression levels of glutathione peroxidase 4 (GPX4), rendering the knock-out cells more sensitive to ferroptosis.
Pre-operative sera interleukin-6 in the diagnosis of high-grade serous ovarian cancer
Authors: Kampan Nirmala Chandralega , Madondo Mutsa Tatenda , Reynolds John , Hallo Julene , McNally Orla M. , Jobling Thomas W. , Stephens Andrew N. , Quinn Michael A. , Plebanski Magdalena
Published: February 2020

Pre-operative discrimination of malignant masses is crucial for accurate diagnosis and prompt referral to a gynae oncology centre for optimal surgical intervention. HGSOC progression is correlated with local and systemic inflammation. We hypothesised that inclusion of inflammatory biomarkers in sera may improve diagnostic tests. In the training cohort, we tested four existing clinical tests (RMI score and ROMA, CA125 and HE4) and a panel of 28 immune soluble biomarkers in sera from 66 patients undergoing surgery for suspected ovarian cancer. Six promising immune biomarkers alone, or in combination with conventional tests, were subsequently analysed in an independent validation cohort (n = 69). IL-6 was identified as the main driver of variability followed closely by conventional diagnostic tests. Median sera IL-6 was higher in HGSOC patients compared to those with a benign mass or controls with normal ovaries (28.3 vs 7.3 vs 1.2 pg/ml, p < 0.0001). The combination of IL-6 further improved the overall predictive probability of the conventional tests. Modelling a two-step triage of women with a suspicious ovarian mass, with IL-6 > 3.75 pg/ml as primary triage followed by conventional tests (CA125 or RMI score) identified ovarian cancer in patients with a misclassification rate of 4.54–3.03%, superior to the use of CA125 or RMI alone (9.09 to 10.60). The validation cohort demonstrated a similar improvement in the diagnostic sensitivity following addition of IL-6. IL-6 in combination with conventional tests may be a useful clinical biomarker for triage of patients with a suspected malignant ovarian mass.
The effect of group size, age and handling frequency on inter-male aggression in CD 1 mice
Authors: Jirkof Paulin , Bratcher Natalie , Medina Letty , Strasburg Donna , Ebert Paige , Gaskill Brianna N.
Published: February 2020

Aggression in male mice often leads to injury and death, making social housing difficult. We tested whether (1) small group size, (2) early age of allocation to a group decreases aggression and 3) manipulation increases aggression in male mice. A 14wk study was performed to assess the following conditions in male CD-1/ICR mice: group size (1, 2, or 3), age at grouping (5 or 7wks), and manipulation (daily scruffing or minimal weekly handling). Wounds, body weights, food consumption, nest scores, sucrose consumption, fecal corticosterone and blood for hematology were collected. At the end of the study, mice were euthanized and pelted to assess wounding with the pelt aggression lesion scale (PALS). No signs of acute or chronic stress were observed in any of the groups. Trio housed mice showed less bite wounds than pair housed mice. In general, mice in larger groups ate less but weighed more. Individually housed mice, however, had high nest scores, low body weights, and increased sucrose and food consumption. These results suggest that even when nesting material is provided, individual mice may be experiencing thermal stress. Based on this data, CD-1 mice can successfully be housed for up to 14wks and groups of 3 may be the best for reducing even minor levels of aggression (i.e. wounding).
Probing nanomechanical responses of cell membranes
Authors: Kim Jichul
Published: February 2020

Despite the importance in various cellular processes, the nanomechanical responses of the living cell membrane have been elusive due to complexities in the membrane associated with the hidden architecture of multiple molecular components, including the lipid bilayer. Here, combined experimental and theoretical frameworks that can probe and interpret nanomechanical responses of the cell membrane are demonstrated. A magnetic tweezer assay was introduced to apply pico-Newton scale forces to lipids and E-cadherin molecules at the living cell surface. Two unique classes of force-extension curves were identified: one with a deflection transition (Type I) and another with a discontinuous transition (Type II). The repeated observations of these responses, regardless of cell type and targeted cell surface molecule, suggest the Type I and II curves are the primary nanomechanical responses of cell membranes. To reproduce these responses in vitro, a model system using synthetic lipid vesicles was also developed. Together with a finite element model of lipid bilayers, the reproduced responses suggest that the confined fluidity and curvature constraints imposed on the lipid bilayer components of the cell membrane are the main parameters responsible for the generation of these responses. This work provides an insight into how forces on membrane molecules propagate to the lipid bilayer components to generate specific nanomechanical responses. In addition, the consistent results obtained using different methodologies demonstrate that the presented force-probing assays and the theoretical model can serve a combined testbed to investigate nanoscale mechanics of the living cell membrane.
Enhanced pre-pubertal nutrition upregulates mitochondrial function in testes and sperm of post-pubertal Holstein bulls
Authors: Johnson Chinju , Dance Alysha , Kovalchuk Igor , Kastelic John , Thundathil Jacob
Published: February 2020

Supplemental energy and protein during calf-hood (2–30 wk) in dairy bulls hastened puberty (~1 mo), upregulated steroid biosynthesis, concentrations of reproductive hormones and Sertoli cell maturation, with larger testes and greater sperm production (~25%) in mature bulls. The objective was to evaluate effects of feeding high (20.0% crude protein [CP], 67.9% total digestible nutrients [TDN]), control/medium (17.0% CP, 66.0% TDN) and low (12.2% CP, 62.9% TDN) diets from 2 to 30 wk on post-pubertal testes of Holstein bulls. Based on RNA sequencing, 497 and 2961 genes were differentially expressed (P < 0.1) in high- vs low- and high- vs medium-diet groups, respectively. According to KEGG analysis, oxidative phosphorylation and ribosome pathways were upregulated in high- vs medium- and low-diet groups, with majority of upregulated genes encoding for essential subunits of complex I, III, IV and V of OXYPHOS pathway. In addition, mitochondrial translation, mitotic nuclear division and cell division were enriched in high- vs medium-diet groups. Consistent with these results, a greater percentage of sperm from high-diet bulls were progressively motile and had normal mitochondrial function compared to medium-diet sperm (P < 0.1). Thus, enhanced early life nutrition upregulated mitochondrial function in testes and sperm of post-pubertal Holstein bulls.
Modification of the head proteome of nurse honeybees (Apis mellifera) exposed to field-relevant doses of pesticides
Authors: Zaluski Rodrigo , Bittarello Alis Correia , Vieira José Cavalcante Souza , Braga Camila Pereira , Padilha Pedro de Magalhaes , Fernandes Mileni da Silva , Bovi Thaís de Souza , Orsi Ricardo de Oliveira
Published: February 2020

Understanding the effect of pesticides on the survival of honeybee colonies is important because these pollinators are reportedly declining globally. In the present study, we examined the changes in the head proteome of nurse honeybees exposed to individual and combined pesticides (the fungicide pyraclostrobin and the insecticide fipronil) at field-relevant doses (850 and 2.5 ppb, respectively). The head proteomes of bees exposed to pesticides were compared with those of bees that were not exposed, and proteins with differences in expression were identified by mass spectrometry. The exposure of nurse bees to pesticides reduced the expression of four of the major royal jelly proteins (MRJP1, MRJP2, MRJP4, and MRJP5) and also several proteins associated with carbohydrate metabolism and energy synthesis, the antioxidant system, detoxification, biosynthesis, amino acid metabolism, transcription and translation, protein folding and binding, olfaction, and learning and memory. Overall, when pyraclostrobin and fipronil were combined, the changes in protein expression were exacerbated. Our results demonstrate that vital proteins and metabolic processes are impaired in nurse honeybees exposed to pesticides in doses close to those experienced by these insects in the field, increasing their susceptibility to stressors and affecting the nutrition and maintenance of both managed and natural colonies.
Alkyladenine DNA glycosylase deficiency uncouples alkylation-induced strand break generation from PARP-1 activation and glycolysis inhibition
Authors: Alhumaydhi Fahad A. , de O. Lopes Debora , Bordin Diana L. , Aljohani Abdullah S. M. , Lloyd Cameron B. , McNicholas Michael D. , Milano Larissa , Charlier Clara F. , Villela Izabel , Henriques João Antonio P. , Plant Kathryn E. , Elliott Ruan M. , Meira Lisiane B.
Published: February 2020

DNA alkylation damage is repaired by base excision repair (BER) initiated by alkyladenine DNA glycosylase (AAG). Despite its role in DNA repair, AAG-initiated BER promotes cytotoxicity in a process dependent on poly (ADP-ribose) polymerase-1 (PARP-1); a NAD+-consuming enzyme activated by strand break intermediates of the AAG-initiated repair process. Importantly, PARP-1 activation has been previously linked to impaired glycolysis and mitochondrial dysfunction. However, whether alkylation affects cellular metabolism in the absence of AAG-mediated BER initiation is unclear. To address this question, we temporally profiled repair and metabolism in wild-type and Aag−/− cells treated with the alkylating agent methyl methanesulfonate (MMS). We show that, although Aag−/− cells display similar levels of alkylation-induced DNA breaks as wild type, PARP-1 activation is undetectable in AAG-deficient cells. Accordingly, Aag−/− cells are protected from MMS-induced NAD+ depletion and glycolysis inhibition. MMS-induced mitochondrial dysfunction, however, is AAG-independent. Furthermore, treatment with FK866, a selective inhibitor of the NAD+ salvage pathway enzyme nicotinamide phosphoribosyltransferase (NAMPT), synergizes with MMS to induce cytotoxicity and Aag−/− cells are resistant to this combination FK866 and MMS treatment. Thus, AAG plays an important role in the metabolic response to alkylation that could be exploited in the treatment of conditions associated with NAD+ dysregulation.
The endocannabinoid hydrolase FAAH is an allosteric enzyme
Authors: Dainese Enrico , Oddi Sergio , Simonetti Monica , Sabatucci Annalaura , Angelucci Clotilde B. , Ballone Alice , Dufrusine Beatrice , Fezza Filomena , De Fabritiis Gianni , Maccarrone Mauro
Published: February 2020

Fatty acid amide hydrolase (FAAH) is a membrane-bound homodimeric enzyme that in vivo controls content and biological activity of N-arachidonoylethanolamine (AEA) and other relevant bioactive lipids termed endocannabinoids. Parallel orientation of FAAH monomers likely allows both subunits to simultaneously recruit and cleave substrates. Here, we show full inhibition of human and rat FAAH by means of enzyme inhibitors used at a homodimer:inhibitor stoichiometric ratio of 1:1, implying that occupation of only one of the two active sites of FAAH is enough to fully block catalysis. Single W445Y substitution in rat FAAH displayed the same activity as the wild-type, but failed to show full inhibition at the homodimer:inhibitor 1:1 ratio. Instead, F432A mutant exhibited reduced specific activity but was fully inhibited at the homodimer:inhibitor 1:1 ratio. Kinetic analysis of AEA hydrolysis by rat FAAH and its F432A mutant demonstrated a Hill coefficient of ~1.6, that instead was ~1.0 in the W445Y mutant. Of note, also human FAAH catalysed an allosteric hydrolysis of AEA, showing a Hill coefficient of ~1.9. Taken together, this study demonstrates an unprecedented allosterism of FAAH, and represents a case of communication between two enzyme subunits seemingly controlled by a single amino acid (W445) at the dimer interface. In the light of extensive attempts and subsequent failures over the last decade to develop effective drugs for human therapy, these findings pave the way to the rationale design of new molecules that, by acting as positive or negative heterotropic effectors of FAAH, may control more efficiently its activity.
Microfabrication of re-entrant surface with hydrophobicity/oleophobicity for liquid foods
Authors: Yamaguchi Masaki
Published: February 2020

Re-entrant texturing may potentially improve the hydrophobicity and oleophobicity of a surface. The food industry requires a microfabrication method to keep surfaces clean without leaving a packaging residue for applications such as food bottles, food containers, and preservation bags. The goal of this study is thus to establish a microfabrication method for re-entrant texturing with spherical curvature to produce hydrophobic/oleophobic surfaces for liquid foods, such as soy sauce and canola oil. Samples with a spherical curvature are created from an ultra-violet-cure (UV-cure) resin and poly (tetrafluoroethylene) (PTFE) microbeads with diameters between 2.26 to 1,353 microns by spin coating on a glass substrate. The resin thickness, the mass and diameter of the microbeads, and the spin coater rotation speed are used as the microfabrication parameters. A side view of samples showing the spherical curvature reveals that a re-entrant texture indeed forms. Distilled water, soy sauce, and canola oil are dropped softly onto the re-entrant surface, however, the droplets cannot be placed stably. For appropriate microbead diameters, the apparent contact angles of soy sauce and canola oil showed 130.2 and 119.4 degrees, respectively. This facile fabrication method for re-entrant surfaces could prove useful for generating hydrophobic/oleophobic surfaces for Newtonian liquid foods.
Melatonin Protects Rabbit Somatic Cell Nuclear Transfer (SCNT) Embryos from Electrofusion Damage
Authors: Qu Pengxiang , Shen Chong , Du Yue , Qin Hongyu , Luo Shiwei , Fu Sixin , Dong Yue , Guo Shuaiqingying , Hu Fang , Xue Ying , Liu Enqi
Published: February 2020

The study’s objectives were to examine the effects of electrofusion on rabbit somatic cell nuclear transfer (SCNT) embryos, and to test melatonin as a protective agent against electrofusion damage to SCNT embryos. The levels of reactive oxygen species (ROS), the epigenetic state (H3K9me3), and the content of endoplasmic reticulum (ER) stress-associated transcripts (IRE-1 and CHOP) were measured. Melatonin was added during the preimplantation development period. The total blastocyst cell numbers were counted, and the fragmentation rate and apoptotic index were determined and used to assess embryonic development. Electrofusion increased (1) ROS levels at the 1-, 2-, 4-, and 8-cell stages; (2) H3K9me3 levels at the 2-, 4-, and 8-cell stage; and (3) the expression of IRE-1 and CHOP at the 8-cell, 16-cell, morula, and blastocyst stages. The treatment of SCNT embryos with melatonin significantly reduced the level of ROS and H3K9me3, and the expression levels of IRE-1 and CHOP. This treatment also significantly reduced the fragmentation rate and apoptotic index of blastocysts and increased their total cell number. In conclusion, the electrofusion of rabbit SCNT embryos induced oxidative stress, disturbed the epigenetic state, and caused ER stress, while melatonin reduced this damage. Our findings are of signal importance for improving the efficiency of SCNT and for optimizing the application of electrical stimulation in other biomedical areas.
Comparative Clinicopathologic Characteristics and Outcomes of Paediatric and Adult Xp11 Translocation Renal Cell Carcinomas: a Retrospective Multicentre Study in China
Authors: Ma Wenliang , Liu Ning , Zhuang Wenyuan , Li Weijian , Qu Feng , Sun Jing , Xu Wei , Zhang Lihua , Jia Ruipeng , Xu Linfeng , Zhao Xiaozhi , Li Xiaogong , Zhang Gutian , Guo Hongqian , Li Dongmei , Gan Weidong
Published: February 2020

This study aimed to compare the clinicopathologic features and prognosis in patients with Xp11 translocation renal cell carcinomas (RCCs). In total, 8083 RCCs were screened at five centres from January 2007 to December 2018, including 8001 adults (≥18 years) and 82 children (<18 years). Finally, 73 adults and 17 children were identified as Xp11 translocation RCCs, accounting for 1.1% (90 of 8083) of the RCCs. However, 4 children and 1 adult were excluded because of loss to follow-up when performing the survival analysis. The proportion of paediatric and adult Xp11 translocation RCCs was 20.7% (17 of 82) and 0.9% (73 of 8001) of RCCs, respectively, and the incidence in children and adults was significantly different (P < 0.01). Lymph node positivity (LN+) most commonly occurred in children (58.8%) compared with adults (28.8%; P = 0.02), but children with LN+ showed significantly higher five-year overall survival and progression-free rates (OS: 75.0%; PFS: 64.8%) than adult patients (OS: 40.3%; PFS: 0%) (log-rank PPFS < 0.01; POS = 0.04). Multivariable analysis indicated that local lymph node metastasis was associated with both PFS (HR = 0.10; 95% CI 0.02–0.51; P = 0.01) and OS (HR = 0.11; 95% CI 0.01–0.98; P = 0.04) in adults. Adult patients with LN+ may indicate a worse prognosis than paediatric patients.
Physiological mechanism underlying the effect of high temperature during anthesis on spikelet-opening of photo-thermo-sensitive genic male sterile rice lines
Authors: Chen Jing , Xu Yangdong , Fei Keqi , Wang Rui , He Jiang , Fu Lidong , Shao Shimei , Li Ke , Zhu Kuanyu , Zhang Weiyang , Wang Zhiqin , Yang Jianchang
Published: February 2020

Decrease in the grain yield resulted from a low percentage of opened spikelets under high temperature (HT) during anthesis is a serious problem in the seed production of photo-thermo-sensitive genic male sterile (PTGMS) rice (Oryza sativaL.) lines, and the mechanism is little understood. Elucidating the physiological mechanism underlying the effect of HT during anthesis on spikelet-opening of PTGMS lines would have great significance in exploring the effective way to mitigate the adverse effect of HT. In this study, two PTGMS lines and one restorer line of rice were used and were subjected to normal temperature (NT) and HT treatments. The results showed that, compared with NT, HT significantly decreased the percentage of opened spikelets, fertilization percentage and seed-setting by significantly increasing the percentage of wrapped spikelets and reducing the spikelet-opening angle, length of spikelet-opening time. The HT significantly decreased the contents of soluble sugars, jasmonic acid (JA) and methyl jasmonate (MeJA) in the lodicules before and at glume-opening, which were significantly correlated with and accounts for the low percentage of opened spikelets under HT for rice, especially for the PTGMS lines.
Target prediction and validation of microRNAs expressed from FSHR and aromatase genes in human ovarian granulosa cells
Authors: Rooda Ilmatar , Hensen Kati , Kaselt Birgitta , Kasvandik Sergo , Pook Martin , Kurg Ants , Salumets Andres , Velthut-Meikas Agne
Published: February 2020

MicroRNAs (miRNAs) are known post-transcriptional regulators of various biological processes including ovarian follicle development. We have previously identified miRNAs from human pre-ovulatory ovarian granulosa cells that are expressed from the intronic regions of two key genes in normal follicular development: FSH receptor (FSHR) andCYP19A1, the latter encoding the aromatase enzyme. The present study aims to identify the target genes regulated by these miRNAs: hsa-miR-548ba and hsa-miR-7973, respectively. The miRNAs of interest were transfected into KGN cell line and the gene expression changes were analyzed by Affymetrix microarray. Potential miRNA-regulated genes were further filtered by bioinformatic target prediction algorithms and validated for direct miRNA:mRNA binding by luciferase reporter assay.LIFR,PTEN,NEO1andSP110were confirmed as targets for hsa-miR-548ba. Hsa-miR-7973 target genesADAM19,PXDNandFMNL3also passed all verification steps. Additionally, the expression pattern of the miRNAs was studied in human primary cumulus granulosa cell culture in relation to the expression of their host genes and FSH stimulation. Based on our findings we propose the involvement of hsa-miR-548ba in the regulation of follicle growth and activation via LIFR and PTEN. Hsa-miR-7973 may be implicated in the modulation of extracellular matrix and cell-cell interactions by regulating the expression of its identified targets.
Gene expression in immortalized versus primary isolated cardiac endothelial cells
Authors: Deng Lisa , Pollmeier Luisa , Zhou Qian , Bergemann Stella , Bode Christoph , Hein Lutz , Lother Achim
Published: February 2020

Endothelial cells take pivotal roles in the heart and the vascular system and their differentiation, subspecification and function is determined by gene expression. A stable, in vitro cardiac endothelial cell line could provide high cell numbers as needed for many epigenetic analyses and facilitate the understanding of molecular mechanisms involved in endothelial cell biology. To test their suitability for transcriptomic or epigenetic studies, we compared the transcriptome of cultured immortalized mouse cardiac endothelial cells (MCEC) to primary cardiac endothelial cells (pEC). Whole transcriptome comparison of MCEC and pEC showed a correlation of 0.75–0.77. Interestingly, correlation of gene expression declined in endothelial cell-typical genes. In MCEC, we found a broad downregulation of genes that are highly expressed in pEC, including well-described markers of endothelial cell differentiation. Accordingly, systematic analysis revealed a downregulation of genes associated with typical endothelial cell functions in MCEC, while genes related to mitotic cell cycle were upregulated when compared to pEC. In conclusion, the findings from this study suggest that primary cardiac endothelial cells should preferably be used for genome-wide transcriptome or epigenome studies. The suitability of in vitro cell lines for experiments investigating single genes or signaling pathways should be carefully validated before use.
Double and Triple Ionisation of Isocyanic Acid
Authors: Eland J. H. D. , Squibb R. J. , Sterling A. J. , Wallner M. , Roos A. Hult , Andersson J. , Axelsson V. , Johansson E. , Teichter A. , Stranges S. , Brunetti B. , Dyke J. M. , Duarte F. , Feifel R.
Published: February 2020

Double and triple ionisation spectra of the reactive molecule isocyanic acid (HNCO) have been measured using multi-electron and ion coincidence techniques combined with synchrotron radiation and compared with high-level theoretical calculations. Vertical double ionisation at an energy of 32.8 ± 0.3 eV forms the 3A” ground state in which the HNCO2+ ion is long lived. The vertical triple ionisation energy is determined as 65 ± 1 eV. The core-valence double ionisation spectra resemble the valence photoelectron spectrum in form, and their main features can be understood on the basis of a simple and rather widely applicable Coulomb model based on the characteristics of the molecular orbitals from which electrons are removed. Characteristics of the most important dissociation channels are examined and discussed.
Single-pixel imaging using caustic patterns
Authors: Toninelli Ermes , Stellinga Daan , Sephton Bereneice , Forbes Andrew , Padgett Miles J.
Published: February 2020

Single-pixel imaging uses a time-varying transmission mask placed in the illumination to achieve imaging without the use of detector arrays. While most research in this field uses sophisticated masks implemented using spatial light modulators, such methods are not available at all lengthscales and wavelengths of illumination. Here we show that alternatively a sequence of projected caustic intensity patterns can be used as the basis for the single-pixel imaging of objects. Caustics can be formed using slowly varying random phase masks, such as for example the surface of a swimming pool, which potentially makes using caustics an option at a range of lengthscales and wavelengths.
Enhanced Dy3+ white emission via energy transfer in spherical (Lu,Gd)3Al5O12 garnet phosphors
Authors: Li Jinkai , Wang Wenzhi , Liu Bin , Duan Guangbin , Liu Zongming
Published: February 2020

The Dy3+doped (Lu,Gd)3Al5O12garnet phosphors with spherical morphology were obtained via homogeneous precipitation method, followed by calcination at 1100 °C. The particle morphology does not change significantly, but can be controlled by adjusting the urea content. The synthesis, structure, luminescent properties of precursor and resultant particles were analyzed by the combined technologies of XRD, FE-SEM, PLE/PL decay behavior. The (Lu0.975Dy0.025)AG phosphors display strong blue and yellow emission at ~481 nm (4F9/2 → 6H15/2transition of Dy3+) and ~582 nm4F9/2 → 6H13/2transition of Dy3+), respectively. The phosphors have similar color coordinate and temperature of (~0.33, ~0.34), ~5517 K, respectively, which are closed to the white emission. The particle size and luminescent intensity decreased while the lifetime increased with the urea concentration increasing. The Gd3+addition does not alter the shape/position of emission peaks, but enhance the blue and yellow emission of Dy3+owing to the efficient Gd3+ → Dy3+energy transfer. The [(Lu1-xGdx)0.975Dy0.025]3Al5O12phosphors are expected to be widely used in the lighting and display areas.
Endoplasmic reticulum stress promotes inflammation-mediated proteolytic activity at the ocular surface
Authors: Woodward Ashley M. , Di Zazzo Antonio , Bonini Stefano , Argüeso Pablo
Published: February 2020

A growing body of evidence implicates endoplasmic reticulum (ER) stress in the pathogenesis of chronic inflammatory and autoimmune disorders. Here, we demonstrate that the proinflammatory cytokine TNFα stimulates matrix metalloproteinase 9 (MMP9) at the ocular surface through a c-Fos-dependent mechanism of ER stress. We found positive reactivity of the molecular chaperone BiP/GRP78 in conjunctival epithelium of patients with ocular cicatricial pemphigoid and increased levels of BiP/GRP78, sXBP1 and GRP94 in human corneal epithelial cells treated with TNFα. Pharmacological blockade of ER stress in vitro using dexamethasone or the chemical chaperones TUDCA and 4PBA attenuated MMP9 expression and secretion in the presence of TNFα. Moreover, expression analysis of genes associated with inflammation and autoimmunity identified the c-Fos proto-oncogene as a mediator of ER stress responses in epithelial cells. Substantially less TNFα-induced MMP9 expression occurred when c-Fos signaling was suppressed with a function-blocking antibody. Taken together, these results indicate that activation of ER stress contributes to promote inflammation-mediated proteolytic activity and uncovers a target for restoring tissue homeostasis in ocular autoimmune disease.
Organosilicons of different molecular size and chemical structure as consolidants for waterlogged archaeological wood – a new reversible and retreatable method
Authors: Broda Magdalena , Dąbek Izabela , Dutkiewicz Agnieszka , Dutkiewicz Michał , Popescu Carmen-Mihaela , Mazela Bartłomiej , Maciejewski Hieronim
Published: February 2020

Ineffectiveness of the chemicals applied so far for waterlogged wood conservation created the need to develop new more, efficient and reliable agents. As an alternative, a new method with the use of organosilicon compounds differing in chemical composition and molecular weight has been investigated. The results obtained show the potential of organosilicons as consolidants in waterlogged wood conservation able to effectively stabilise wood dimensions upon drying. The best wood stabilisers were low-molecular organosilicons enable to penetrate the cell wall as well as chemicals with functional groups capable of interacting with wood polymers and forming stabilising coatings on the cell wall surface. The best anti-shrink efficiency values were obtained for (3-Mercaptopropyl)trimethoxysilane, (3-Aminopropyl)triethoxysilane, 1,3-Bis(3-aminopropyl)-1,1,3,3-tetramethyldisiloxane, reaching 98, 91 and 91%, respectively. Most of the applied organosilicons reduced wood hygroscopicity, which limits the risk of further dimensional changes of wood exposed to a variable air moisture content and potentially reduces wood biodegradation. In the light of our studies, the proposed method of waterlogged wood conservation with organosilicons is potentially reversible in the case of siloxanes and amino-silanes as well as retreatable, which complies with the requirements of the conservation ethics.
Effect of mild blast-induced TBI on dendritic architecture of the cortex and hippocampus in the mouse
Authors: Ratliff Whitney A. , Mervis Ronald F. , Citron Bruce A. , Schwartz Brian , Rubovitch Vardit , Schreiber Shaul , Pick Chaim G.
Published: February 2020

Traumatic brain injury (TBI) has been designated as a signature injury of modern military conflicts. Blast trauma, in particular, has come to make up a significant portion of the TBIs which are sustained in warzones. Though most TBIs are mild, even mild TBI can induce long term effects, including cognitive and memory deficits. In our study, we utilized a mouse model of mild blast-related TBI (bTBI) to investigate TBI-induced changes within the cortex and hippocampus. We performed rapid Golgi staining on the layer IV and V pyramidal neurons of the parietal cortex and the CA1 basilar tree of the hippocampus and quantified dendritic branching and distribution. We found decreased dendritic branching within both the cortex and hippocampus in injured mice. Within parietal cortex, this decreased branching was most evident within the middle region, while outer and inner regions resembled that of control mice. This study provides important knowledge in the study of how the shockwave associated with a blast explosion impacts different brain regions.
Double obstacles increase gait asymmetry during obstacle crossing in people with Parkinson’s disease and healthy older adults: A pilot study
Authors: Orcioli-Silva Diego , Barbieri Fabio Augusto , dos Santos Paulo Cezar Rocha , Beretta Victor Spiandor , Simieli Lucas , Vitorio Rodrigo , Lirani-Silva Ellen , Gobbi Lilian Teresa Bucken
Published: February 2020

Gait asymmetry during unobstructed walking in people with Parkinson’s disease (PD) has been well documented. However, under complex situations, such as environments with double obstacles, gait asymmetry remains poorly understood in PD. Therefore, the aim of this study was to analyze inter-limb asymmetry while crossing a single obstacle and double obstacles (with different distances between them) in people with PD and healthy older adults. Nineteen people with PD and 19 healthy older people performed three conditions: (i) walking with one obstacle (Single); (ii) walking with two obstacles with a 50 cm distance between them (Double-50); (iii) walking with two obstacles with a 108 cm distance between them (Double-108). The participants performed the obstacle crossing with both lower limbs. Asymmetry Index was calculated. We found that people with PD presented higher leading and trailing toe clearance asymmetry than healthy older people. In addition, participants increased asymmetry in the Double-50 compared to Single condition. It can be concluded that people with PD show higher asymmetry during obstacle crossing compared to healthy older people, independently of the number of obstacles. In addition, a challenging environment induces asymmetry during obstacle crossing in both people with PD and healthy older people.