bluuebunny/crossref_metadata_2025 huggingface dataset:
PPARα-Mediated Positive-Feedback Loop Contributes to Cold Exposure Memory
Authors: Alfaqaan Soaad , Yoshida Tomoki , Imamura Hiromi , Tsukano Chihiro , Takemoto Yoshiji , Kakizuka Akira
Published: March 2019

Fluctuations in food availability and shifts in temperature are typical environmental changes experienced by animals. These environmental shifts sometimes portend more severe changes; e.g., chilly north winds precede the onset of winter. Such telltale signs may be indicators for animals to prepare for such a shift. Here we show that HEK293A cells, cultured under starvation conditions, can “memorize” a short exposure to cold temperature (15 °C), which was evidenced by their higher survival rate compared to cells continuously grown at 37 °C. We refer to this phenomenon as “cold adaptation”. The cold-exposed cells retained high ATP levels, and addition of etomoxir, a fatty acid oxidation inhibitor, abrogated the enhanced cell survival. In our standard protocol, cold adaptation required linoleic acid (LA) supplementation along with the activity of Δ-6-desaturase (D6D), a key enzyme in LA metabolism. Moreover, supplementation with the LA metabolite arachidonic acid (AA), which is a high-affinity agonist of peroxisome proliferator-activated receptor-alpha (PPARα), was able to underpin the cold adaptation, even in the presence of a D6D inhibitor. Cold exposure with added LA or AA prompted a surge in PPARα levels, followed by the induction of D6D expression; addition of a PPARα antagonist or a D6D inhibitor abrogated both their expression, and reduced cell survival to control levels. We also found that the brief cold exposure transiently prevents PPARα degradation by inhibiting the ubiquitin proteasome system, and starvation contributes to the enhancement of PPARα activity by inhibiting mTORC1. Our results reveal an innate adaptive positive-feedback mechanism with a PPARα-D6D-AA axis that is triggered by a brief cold exposure in cells. “Cold adaptation” could have evolved to increase strength and resilience against imminent extreme cold temperatures.
Somatic symptom load in men and women from middle to high age in the Gutenberg Health Study - association with psychosocial and somatic factors
Authors: Beutel Manfred E. , Wiltink Jörg , Ghaemi Kerahrodi Jasmin , Tibubos Ana N. , Brähler Elmar , Schulz Andreas , Wild Philipp , Münzel Thomas , Lackner Karl , König Jochem , Pfeiffer Norbert , Michal Matthias , Henning Michaela
Published: March 2019

The purpose of the study was (1) to determine the prevalence of somatic symptoms in men and women in the general population and (2) to identify the contributions of psychosocial factors and somatic disease on symptom reporting. A total of 7,925 participants aged 40 to 80 years underwent medical and psychological assessments, based on the PHQ-15 (Patient Health Questionnaire). We excluded 3 items in order to avoid confounding findings: 2 items overlapping with the depression measure (PHQ-9) and the menstruation complaints item which biases sex comparisons. Pain complaints (arms, legs, joints, back pain) affected the majority of men and women, and somatic symptom reporting increased with age. When confounding has been reduced, psychosocial factors (lack of social support, adverse life events, loneliness, depression, generalized anxiety, panic, social phobia) have remained the strongest predictors of somatic symptoms. As shown by the interaction between sex and depression, depression plays a smaller role for somatic symptom reporting in women vs. men. Findings highlight the complex psychosocial and somatic contributions to somatic symptom reporting.
Highly interacting regions of the human genome are enriched with enhancers and bound by DNA repair proteins
Authors: Sobhy Haitham , Kumar Rajendra , Lewerentz Jacob , Lizana Ludvig , Stenberg Per
Published: March 2019

In specific cases, chromatin clearly forms long-range loops that place distant regulatory elements in close proximity to transcription start sites, but we have limited understanding of many loops identified by Chromosome Conformation Capture (such as Hi-C) analyses. In efforts to elucidate their characteristics and functions, we have identified highly interacting regions (HIRs) using intra-chromosomal Hi-C datasets with a new computational method based on looking at the eigenvector that corresponds to the smallest eigenvalue (here unity). Analysis of these regions using ENCODE data shows that they are in general enriched in bound factors involved in DNA damage repair and have actively transcribed genes. However, both highly transcribed regions as well as transcriptionally inactive regions can form HIRs. The results also indicate that enhancers and super-enhancers in particular form long-range interactions within the same chromosome. The accumulation of DNA repair factors in most identified HIRs suggests that protection from DNA damage in these regions is essential for avoidance of detrimental rearrangements.
Analyses of 202 plastid genomes elucidate the phylogeny of Solanum section Petota
Authors: Huang Binquan , Ruess Holly , Liang Qiqi , Colleoni Christophe , Spooner David M.
Published: March 2019

Our paper analyzes full plastid DNA sequence data of 202 wild and cultivated diploid potatoes,SolanumsectionPetota, to explore its phylogenetic utility compared to prior analyses of the same accessions using genome-wide nuclear SNPs, and plastid DNA restriction site data. The present plastid analysis discovered the same major clades as the nuclear data but with some substantial differences in topology within the clades. The considerably larger plastid and nuclear data sets add phylogenetic resolution within the prior plastid DNA restriction site data, highlight plastid/nuclear incongruence that supports hypotheses of hybridization/introgression to help explain the taxonomic difficulty in the section.
Rapid identification and phylogenetic classification of diverse bacterial pathogens in a multiplexed hybridization assay targeting ribosomal RNA
Authors: Bhattacharyya Roby P. , Walker Mark , Boykin Rich , Son Sophie S. , Liu Jamin , Hachey Austin C. , Ma Peijun , Wu Lidan , Choi Kyungyong , Cummins Kaelyn C. , Benson Maura , Skerry Jennifer , Ryu Hyunryul , Wong Sharon Y. , Goldberg Marcia B. , Han Jongyoon , Pierce Virginia M. , Cosimi Lisa A. , Shoresh Noam , Livny Jonathan , Beechem Joseph , Hung Deborah T.
Published: March 2019

Rapid bacterial identification remains a critical challenge in infectious disease diagnostics. We developed a novel molecular approach to detect and identify a wide diversity of bacterial pathogens in a single, simple assay, exploiting the conservation, abundance, and rich phylogenetic content of ribosomal RNA in a rapid fluorescent hybridization assay that requires no amplification or enzymology. Of 117 isolates from 64 species across 4 phyla, this assay identified bacteria with >89% accuracy at the species level and 100% accuracy at the family level, enabling all critical clinical distinctions. In pilot studies on primary clinical specimens, including sputum, blood cultures, and pus, bacteria from 5 different phyla were identified.
Single-cell transcriptome analysis identifies distinct cell types and niche signaling in a primary gastric organoid model
Authors: Chen Jiamin , Lau Billy T. , Andor Noemi , Grimes Susan M. , Handy Christine , Wood-Bouwens Christina , Ji Hanlee P.
Published: March 2019

The diverse cellular milieu of the gastric tissue microenvironment plays a critical role in normal tissue homeostasis and tumor development. However, few cell culture model can recapitulate the tissue microenvironment and intercellular signalingin vitro. We used a primary tissue culture system to generate a murine p53 null gastric tissue model containing both epithelium and mesenchymal stroma. To characterize the microenvironment and niche signaling, we used single cell RNA sequencing (scRNA-Seq) to determine the transcriptomes of 4,391 individual cells. Based on specific markers, we identified epithelial cells, fibroblasts and macrophages in initial tissue explants during organoid formation. The majority of macrophages were polarized towards wound healing and tumor promotion M2-type. During the course of time, the organoids maintained both epithelial and fibroblast lineages with the features of immature mouse gastric stomach. We detected a subset of cells in both lineages expressingLgr5, one of the stem cell markers. We examined the lineage-specific Wnt signaling activation, and identified thatRspo3was specifically expressed in the fibroblast lineage, providing an endogenous source of the R-spondin to activate Wnt signaling. Our studies demonstrate that this primary tissue culture system enables one to study gastric tissue niche signaling and immune responsein vitro.
Identification of polysaccharide capsules among extensively drug-resistant genitourinary Haemophilus parainfluenzae isolates
Authors: González-Díaz Aida , Tubau Fe , Pinto Miguel , Sierra Yanik , Cubero Meritxell , Càmara Jordi , Ayats Josefina , Bajanca-Lavado Paula , Ardanuy Carmen , Marti Sara
Published: March 2019

The human commensal Haemophilus parainfluenzae is emerging as an opportunistic multidrug-resistant pathogen. The objectives of this work were to characterise a new capsular operon of extensively drug-resistant (XDR) H. parainfluenzae clinical isolates and study their resistance mechanisms using whole-genome sequencing. All strains were resistant to: ß-lactams, via amino acid changes in PBP3 (S385T, I442F, V511A, N526K and V562I); quinolones, by alterations in GyrA (S84F and D88Y) and ParC (S84F and S138T); chloramphenicol, through the presence of catS; macrolides, via the presence of mel and mef(E)-carrying MEGA element; and tetracycline, through the presence of tet(M) and/or tet(B). Phylogenetic analysis revealed high genomic diversity when compared to the H. parainfluenzae genomes available on the NCBI, the isolates from this study being closely related to the Swiss XDR AE-2096513. A full capsular operon showing homology to that of H. influenzae was identified, in accordance with the observation of a capsular structure by TEM. This study describes for the first time a capsular operon in H. parainfluenzae, a major determinant of pathogenicity that may contribute to increased virulence in XDR clinical isolates. Moreover, phylogenetic analysis suggests the possible spread of an XDR-encapsulated strain in Europe.
An analysis of benign human prostate offers insights into the mechanism of apocrine secretion and the origin of prostasomes
Authors: Fullwood Nigel J. , Lawlor Alan J. , Martin-Hirsch Pierre L. , Matanhelia Shyam S. , Martin Francis L.
Published: March 2019

The structure and function of normal human prostate is still not fully understood. Herein, we concentrate on the different cell types present in normal prostate, describing some previously unreported types and provide evidence that prostasomes are primarily produced by apocrine secretion. Patients (n = 10) undergoing TURP were prospectively consented based on their having a low risk of harbouring CaP. Scanning electron microscopy and transmission electron microscopy was used to characterise cell types and modes of secretion. Zinc levels were determined using Inductively Coupled Plasma Mass Spectrometry. Although merocrine secretory cells were noted, the majority of secretory cells appear to be apocrine; for the first time, we clearly show high-resolution images of the stages of aposome secretion in human prostate. We also report a previously undescribed type of epithelial cell and the first ultrastructural image of wrapping cells in human prostate stroma. The zinc levels in the tissues examined were uniformly high and X-ray microanalysis detected zinc in merocrine cells but not in prostasomes. We conclude that a significant proportion of prostasomes, possibly the majority, are generated via apocrine secretion. This finding provides an explanation as to why so many large proteins, without a signal peptide sequence, are present in the prostatic fluid.
Wet-tip versus dry-tip regimes of osmotically driven fluid flow
Authors: Ostrenko Oleksandr , Hampe Jochen , Brusch Lutz
Published: March 2019

The secretion of osmolytes into a lumen and thereby caused osmotic water inflow can drive fluid flows in organs without a mechanical pump. Such fluids include saliva, sweat, pancreatic juice and bile. The effects of elevated fluid pressure and the associated mechanical limitations of organ function remain largely unknown since fluid pressure is difficult to measure inside tiny secretory channels in vivo. We consider the pressure profile of the coupled osmolyte-flow problem in a secretory channel with a closed tip and an open outlet. Importantly, the entire lateral boundary acts as a dynamic fluid source, the strength of which self-organizes through feedback from the emergent pressure solution itself. We derive analytical solutions and compare them to numerical simulations of the problem in three-dimensional space. The theoretical results reveal a phase boundary in a four-dimensional parameter space separating the commonly considered regime with steady flow all along the channel, here termed “wet-tip” regime, from a “dry-tip” regime suffering ceased flow downstream from the closed tip. We propose a relation between the predicted phase boundary and the onset of cholestasis, a pathological liver condition with reduced bile outflow. The phase boundary also sets an intrinsic length scale for the channel which could act as a length sensor during organ growth.
Chaotic optical power dropouts driven by low frequency bias forcing in a mid-infrared quantum cascade laser
Authors: Spitz Olivier , Wu Jiagui , Carras Mathieu , Wong Chee-Wei , Grillot Frédéric
Published: March 2019

Mid-infrared quantum cascade lasers operating under external optical feedback and external periodic bias forcing are shown to exhibit a deterministic chaotic pattern composed of frequencies which are linked to the one of the forcing. Results also show that both the amplitude and the frequency of the forcing play a key role in the number of retrieved spikes per modulation period. These findings are of paramount importance for chaotic operation of quantum cascade lasers in applications such as optical countermeasure systems and secure atmospheric transmission lines, as well as for simulating neuronal systems and the communication between neurons due to sudden bursts.
Observation of an expert model induces a skilled movement coordination pattern in a single session of intermittent practice
Authors: Friedman Jason , Korman Maria
Published: March 2019

We tested how observation of a skilled pattern of planar movements can assist in the learning of a new motor skill, which otherwise requires rigorous long-term practice to achieve fast and smooth performance. Sixty participants performed a sequence of planar hand movements on pre-test, acquisition, post-test and 24 h post-training blocks, under 1 of 4 conditions: an observation group (OG), a slowed observation group (SOG), a random motion control group (RMCG) and a double physical training control group (DPTCG). The OG and SOG observed an expert model’s right hand performing the study task intermittently throughout acquisition, RMCG observed random dots movement instead of a model. Participants in the DPTCG received extra physical practice trials instead of the visually observed trials. Kinematic analysis revealed that only in conditions with observation of an expert model there was an instant robust improvement in motor planning of the task. This step-wise improvement was not only persistent in post-training retests but was also apparently implicit and subject to further incremental improvements in movement strategy over the period of 24 hours. The rapid change in motor strategy was accompanied by a transient within-session increase in spatial error for the observation groups, but this went away by 24 h post-training. We suggest that observation of hand movements of an expert model coaligned with self-produced movements during training can significantly condense the time-course of ecologically relevant drawing/writing skill mastery.
Improving estimation of puma (Puma concolor) population density: clustered camera-trapping, telemetry data, and generalized spatial mark-resight models
Authors: Murphy Sean M. , Wilckens David T. , Augustine Ben C. , Peyton Mark A. , Harper Glenn C.
Published: March 2019

Obtaining reliable population density estimates for pumas (Puma concolor) and other cryptic, wide-ranging large carnivores is challenging. Recent advancements in spatially explicit capture-recapture models have facilitated development of novel survey approaches, such as clustered sampling designs, which can provide reliable density estimation for expansive areas with reduced effort. We applied clustered sampling to camera-traps to detect marked (collared) and unmarked pumas, and used generalized spatial mark-resight (SMR) models to estimate puma population density across 15,314 km2 in the southwestern USA. Generalized SMR models outperformed conventional SMR models. Integrating telemetry data from collars on marked pumas with detection data from camera-traps substantially improved density estimates by informing cryptic activity (home range) center transiency and improving estimation of the SMR home range parameter. Modeling sex of unmarked pumas as a partially identifying categorical covariate further improved estimates. Our density estimates (0.84–1.65 puma/100 km2) were generally more precise (CV = 0.24–0.31) than spatially explicit estimates produced from other puma sampling methods, including biopsy darting, scat detection dogs, and regular camera-trapping. This study provides an illustrative example of the effectiveness and flexibility of our combined sampling and analytical approach for reliably estimating density of pumas and other wildlife across geographically expansive areas.
Circulating CD56bright NK cells inversely correlate with survival of melanoma patients
Authors: de Jonge Kaat , Ebering Anna , Nassiri Sina , Maby-El Hajjami Hélène , Ouertatani-Sakouhi Hajer , Baumgaertner Petra , Speiser Daniel E.
Published: March 2019

The roles of NK cells in human melanoma remain only partially understood. We characterized NK cells from peripheral blood ex vivo by flow cytometry obtained from late stage (III/IV) melanoma patients. Interestingly, we found that the abundance of CD56bright NK cells negatively correlate with overall patient survival, together with distant metastases, in a multivariate cox regression analysis. The patients’ CD56bright NK cells showed upregulation of CD11a, CD38 and CD95 as compared to healthy controls, pointing to an activated phenotype as well as a possible immune regulatory role in melanoma patients. After stimulation in vitro, CD56bright NK cells produced less TNFα and GMCSF in patients than controls. Furthermore, IFNγ production by the CD56bright NK cells correlated inversely with overall survival. Our results highlight that abundance and function of CD56bright NK cells are associated with melanoma patient survival, emphasizing the potential of NK cell subsets for biomarker discovery and future therapeutic targeting.
‘Genome skimming’ with the MinION hand-held sequencer identifies CITES-listed shark species in India’s exports market
Authors: Johri Shaili , Solanki Jitesh , Cantu Vito Adrian , Fellows Sam R. , Edwards Robert A. , Moreno Isabel , Vyas Asit , Dinsdale Elizabeth A.
Published: March 2019

Chondrichthyes - sharks, rays, skates, and chimeras, are among the most threatened and data deficient vertebrate species. Global demand for shark and ray derived products, drives unregulated and exploitative fishing practices, which are in turn facilitated by the lack of ecological data required for effective conservation of these species. Here, we describe a Next Generation Sequencing method (using the MinION, a hand-held portable sequencing device from Oxford Nanopore Technologies), and analyses pipeline for molecular ecological studies in Chondrichthyes. Using this method, the complete mitochondrial genome and nuclear intergenic and protein-coding sequences were obtained by direct sequencing of genomic DNA obtained from shark fin tissue. Recovered loci include mitochondrial barcode sequences- Cytochrome oxidase I, NADH2, 16S rRNA and 12S rRNA- and nuclear genetic loci such as 5.8S rRNA, Internal Transcribed Spacer 2, and 28S rRNA regions, which are commonly used for taxonomic identification. Other loci recovered were the nuclear protein-coding genes for antithrombin or SerpinC, Immunoglobulin lambda light chain, Preprogehrelin, selenium binding protein 1(SBP1), Interleukin-1 beta (IL-1β) and Recombination-Activating Gene 1 (RAG1). The median coverage across all genetic loci was 20x and sequence accuracy was ≥99.8% compared to reference sequences. Analyses of the nuclear ITS2 region and the mitochondrial protein-encoding loci allowed accurate taxonomic identification of the shark specimen asCarcharhinus falciformis, a CITES Appendix II species. MinION sequencing provided 1,152,211 bp of new shark genome, increasing the number of sequenced shark genomes to five. Phylogenetic analyses using both mitochondrial and nuclear loci provided evidence thatPrionace glaucais nested withinCarcharhinus, suggesting the need for taxonomic reassignment ofP.glauca. We increased genomic information about a shark species for ecological and population genetic studies, enabled accurate identification of the shark tissue for biodiversity indexing and resolved phylogenetic relationships among multiple taxa. The method was independent of amplification bias, and adaptable for field assessments of other Chondrichthyes and wildlife species in the future.
Development and Validation of Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) for Rapid Detection of ZIKV in Mosquito Samples from Brazil
Authors: Silva Severino Jefferson Ribeiro da , Paiva Marcelo Henrique Santos , Guedes Duschinka Ribeiro Duarte , Krokovsky Larissa , Melo Fábio Lopes de , Silva Maria Almerice Lopes da , Silva Adalúcia da , Ayres Constância Flávia Junqueira , Pena Lindomar J.
Published: March 2019

The rapid spread of Zika virus (ZIKV) represents a global public health problem, especially in areas that harbor several mosquito species responsible for virus transmission, such as Brazil. In these areas, improvement in mosquito control needs to be a top priority, but mosquito viral surveillance occurs inefficiently in ZIKV-endemic countries. Quantitative reverse transcription PCR (qRT-PCR) is the gold standard for molecular diagnostic of ZIKV in both human and mosquito samples. However, the technique presents high cost and limitations for Point-of-care (POC) diagnostics, which hampers its application for a large number of samples in entomological surveillance programs. Here, we developed and validated a one-step reverse transcription LAMP (RT-LAMP) platform for detection of ZIKV in mosquito samples. The RT-LAMP assay was highly specific for ZIKV and up to 10,000 times more sensitive than qRT-PCR. Assay validation was performed using 60 samples from Aedes aegypti and Culex quinquefasciatus mosquitoes collected in Pernambuco State, Brazil, which is at the epicenter of the Zika epidemic. The RT-LAMP had a sensitivity of 100%, specificity of 91.18%, and overall accuracy of 95.24%. Thus, our POC diagnostics is a powerful and inexpensive tool to monitor ZIKV in mosquito populations and will allow developing countries to establish better control strategies for this devastating pathogen.
Pathogenetic basis of Takenouchi-Kosaki syndrome: Electron microscopy study using platelets in patients and functional studies in a Caenorhabditis elegans model
Authors: Uehara Tomoko , Suzuki Hidenori , Okamoto Nobuhiko , Kondoh Tatsuro , Ahmad Ayesha , O’Connor Bridget C. , Yoshina Sawako , Mitani Shohei , Kosaki Kenjiro , Takenouchi Toshiki
Published: March 2019

The combined phenotype of thrombocytopenia accompanied by intellectual disability in patients with a de novo heterozygous mutation, i.e., p.Tyr64Cys in CDC42, signifies a clinically recognizable novel syndrome that has been eponymized as “Takenouchi-Kosaki syndrome” (OMIM #616737). In the present study, a detailed phenotypic analysis performed for a total of five patients with Takenouchi-Kosaki syndrome revealed that intellectual disability, macrothrombocytopenia, camptodactyly, structural brain abnormalities with sensorineural deafness, hypothyroidism, and frequent infections comprise the cardinal features of this condition. A morphologic analysis of platelets derived from three affected individuals was performed using electron microscopy. The platelets of the three patients were large and spherical in shape. Furthermore, platelet α-granules were decreased, while vacuoles were increased. We further performed a functional analysis of p.Tyr64Cys in CDC42 through CRISPR/Cas9-mediated gene editing in a Caenorhabditis elegans model. This functional analysis suggested that the mutant allele has hypomorphic effects. Takenouchi-Kosaki syndrome is clinically recognizable by the combined phenotype of intellectual disability, macrothrombocytopenia, camptodactyly, structural brain abnormalities with sensorineural deafness, hypothyroidism, and frequent infections as well as the identification of a heterozygous de novo mutation in CDC42, i.e., p.Tyr64Cys.
Night workers have lower levels of antioxidant defenses and higher levels of oxidative stress damage when compared to day workers
Authors: Teixeira Kely R. C. , dos Santos Camila P. , de Medeiros Luciana A. , Mendes Jordane A. , Cunha Thúlio M. , De Angelis Kátia , Penha-Silva Nilson , de Oliveira Erick P. , Crispim Cibele A.
Published: March 2019

The effects of circadian misalignment and work shift on oxidative stress profile of shift workers have not been explored in the literature. The present study aimed to evaluate the role of shift work (day and night) and social jetlag - a measure of circadian misalignment - with oxidative stress markers. A cross-sectional study was performed with 79 men (21–65 years old, 27.56 ± 4.0 kg/m2) who worked the night shift (n = 37) or daytime (n = 42). The analyzed variables included anthropometric measures and determination of systemic levels of markers of oxidative damage and antioxidant defense. Social jetlag was calculated by the absolute difference between the mean sleep point on working and rest days. The night group presented higher systemic values of thiobarbituric acid reactive substances and hydrogen peroxide, and lower levels of nitrite, total antioxidant capacity, and catalase and superoxide dismutase activities in relation to the day group. However, social jetlag was not associated with oxidative stress-related biomarkers analyzed in the night group. These results suggest that the night worker has higher levels of oxidative stress damage and lower levels of antioxidant defenses, while social jetlag was not a possible responsible factor for this condition.
Systemic LPS-induced Aβ-solubilization and clearance in AβPP-transgenic mice is diminished by heparanase overexpression
Authors: Jendresen Charlotte , Digre Andreas , Cui Hao , Zhang Xiao , Vlodavsky Israel , Li Jin-Ping , Nilsson Lars N. G.
Published: March 2019

Amyloid-β (Aβ) is the main constituent of amyloid deposits in Alzheimer’s disease (AD). The neuropathology is associated with neuroinflammation. Here, we investigated effects of systemic lipopolysaccharide (LPS)-treatment on neuroinflammation and Aβ deposition in AβPP-mice and double-transgenic mice with brain expression of AβPP and heparanase, an enzyme that degrades HS and generates an attenuated LPS-response. At 13 months of age, the mice received a single intraperitoneal injection of 50 µg LPS or vehicle, and were sacrificed 1.5 months thereafter. Aβ in the brain was analyzed histologically and biochemically after sequential detergent extraction. Neuroinflammation was assessed by CD45 immunostaining and mesoscale cytokine/chemokine ELISA. In single-transgenic mice, LPS-treatment reduced total Aβ deposition and increased Tween-soluble Aβ. This was associated with a reduced CXCL1, IL-1β, TNF-α-level and microgliosis, which correlated with amyloid deposition and total Aβ. In contrast, LPS did not change Aβ accumulation or inflammation marker in the double-transgenic mice. Our findings suggest that a single pro-inflammatory LPS-stimulus, if given sufficient time to act, triggers Aβ-clearance in AβPP-transgenic mouse brain. The effects depend on HS and heparanase.
Androgens Induce Invasiveness of Triple Negative Breast Cancer Cells Through AR/Src/PI3-K Complex Assembly
Authors: Giovannelli Pia , Di Donato Marzia , Auricchio Ferdinando , Castoria Gabriella , Migliaccio Antimo
Published: March 2019

Breast cancer (BC) is still characterized by high morbidity and mortality. A specific BC subtype named triple negative BC (TNBC) lacks estrogen and progesterone receptors (ER and PR, respectively) and is characterized by the absence of overexpression/amplification of human epidermal growth factor receptor 2 (HER2). The androgen receptor (AR) is expressed in TNBC, although its function in these cancers is still debated. Moreover, few therapeutic options are currently available for the treatment of TNBC. In this study, we have used TNBC-derived MDA-MB231 and MDA-MB453 cells that, albeit at different extent, both express AR. Androgen challenging induces migration and invasiveness of these cells. Use of the anti-androgen bicalutamide or AR knockdown experiments show that these effects depend on AR. Furthermore, the small peptide, S1, which mimics the AR proline-rich motif responsible for the interaction of AR with SH3-Src, reverses the effects in both cell lines, suggesting that the assembly of a complex made up of AR and Src drives the androgen-induced motility and invasiveness. Co-immunoprecipitation experiments in androgen-treated MDA-MB231 and MDA-MB453 cells show that the AR/Src complex recruits p85α, the regulatory subunit of PI3-K. In such a way, the basic machinery leading to migration and invasiveness is turned-on. The S1 peptide inhibits motility and invasiveness of TNBC cells and disrupts the AR/Src/p85α complex assembly in MDA-MB231 cells. This study shows that the rapid androgen activation of Src/PI3-K signaling drives migration and invasiveness of TNBC cells and suggests that the S1 peptide is a promising therapeutic option for these cancers.
Response of healthy local tomato (Solanum lycopersicum L.) populations to grafting in organic farming
Authors: Moreno Marta María , Villena Jaime , González-Mora Sara , Moreno Carmen
Published: March 2019

Demands for tomato local varieties are increasing worldwide, especially in organic farming mainly for their high sensory value and attractive appearance. This is the case of the “Moruno” tomato type, widely grown in the Mediterranean countries and greatly due to its highly appreciated organoleptic attributes but low yield or a short postharvest period. For this reason, the study aimed to assess if grafting of local “Moruno” populations (Mor-62, Mor-204) using commercial rootstocks (King-Kong F1, K; Multifort F1, M; Spirit F1, S) affects yield, fruit nutritional and functional composition, postharvest storage and consumer acceptance. Results showed differences between both populations, while rootstocks were only different for the glucose content and the flavour quality. Grafting improved the marketable yield (~43%), fruit number (~22%) and mean fruit weight (~12%), but had no incidence on the blossom-end rot disorder. This technique increased the carotenoid (lycopene, β-carotene and total carotene) content but decreased the organic acids (malic and citric) and sugar (fructose and glucose) rates, while ascorbic acid was not affected. The fruit postharvest storage was not practically modified. However, the overall flavour preference and visual appearance varied depending on the scion.
Super-resolution microscopy reveals significant impact of M2e-specific monoclonal antibodies on influenza A virus filament formation at the host cell surface
Authors: Kolpe Annasaheb , Arista-Romero Maria , Schepens Bert , Pujals Silvia , Saelens Xavier , Albertazzi Lorenzo
Published: March 2019

Influenza A virions are highly pleomorphic, exhibiting either spherical or filamentous morphology. The influenza A virus strain A/Udorn/72 (H3N2) produces copious amounts of long filaments on the surface of infected cells where matrix protein 1 (M1) and 2 (M2) play a key role in virus filament formation. Previously, it was shown that an anti-M2 ectodomain (M2e) antibody could inhibit A/Udorn/72 virus filament formation. However, the study of these structures is limited by their small size and complex structure. Here, we show that M2e-specific IgG1 and IgG2a mouse monoclonal antibodies can reduce influenza A/Udorn/72 virus plaque growth and infectivity in vitro. Using Immuno-staining combined with super-resolution microscopy that allows us to study structures beyond the diffraction limit, we report that M2 is localized at the base of viral filaments that emerge from the membrane of infected cells. Filament formation was inhibited by treatment of A/Udorn/72 infected cells with M2e-specific IgG2a and IgG1 monoclonal antibodies and resulted in fragmentation of pre-existing filaments. We conclude that M2e-specific IgGs can reduce filamentous influenza A virus replication in vitro and suggest that in vitro inhibition of A/Udorn/72 virus replication by M2e-specific antibodies correlates with the inhibition of filament formation on the surface of infected cells.
Development of a subsurface LIBS sensor for in situ groundwater quality monitoring with applications in CO2 leak sensing in carbon sequestration
Authors: Hartzler D. A. , Jain J. C. , McIntyre D. L.
Published: March 2019

Sub-surface activity such as geologic carbon sequestration (GCS) has the potential to contaminate groundwater sources with dissolved metals originating from sub-surface brines or leaching of formation rock. Therefore, a Laser Induced Breakdown Spectroscopy (LIBS) based sensor is developed for sub-surface water quality monitoring. The sensor head is built using a low cost passively Q-switched (PQSW) laser and is fiber coupled to a pump laser and a gated spectrometer. The prototype sensor head was constructed using off the shelf components and a custom monolithic, PQSW laser and testing has verified that the fiber coupled design performs as desired. The system shows good calibration linearity for tested elements (Ca, Sr, and K), quick data collection times, and Limits of Detection (LODs) that are comparable to or better than those of table top, actively Q-switched systems. The fiber coupled design gives the ability to separate the PQSW LIBS excitation laser from the pump source and spectrometer, allowing these expensive and fragile components to remain at the surface while only the low-cost, all optical sensor head needs to be exposed to the hostile downhole environment.
Increased FUS levels in astrocytes leads to astrocyte and microglia activation and neuronal death
Authors: Ajmone-Cat Maria Antonietta , Onori Angela , Toselli Camilla , Stronati Eleonora , Morlando Mariangela , Bozzoni Irene , Monni Emanuela , Kokaia Zaal , Lupo Giuseppe , Minghetti Luisa , Biagioni Stefano , Cacci Emanuele
Published: March 2019

Mutations of Fused in sarcoma (FUS), a ribonucleoprotein involved in RNA metabolism, have been found associated with both familial and sporadic cases of amyotrophic lateral sclerosis (ALS). Notably, besides mutations in the coding sequence, also mutations into the 3′ untranslated region, leading to increased levels of the wild-type protein, have been associated with neuronal death and ALS pathology, in ALS models and patients. The mechanistic link between altered FUS levels and ALS-related neurodegeneration is far to be elucidated, as well as the consequences of elevated FUS levels in the modulation of the inflammatory response sustained by glial cells, a well-recognized player in ALS progression. Here, we studied the effect of wild-type FUS overexpression on the responsiveness of mouse and human neural progenitor-derived astrocytes to a pro-inflammatory stimulus (IL1β) used to mimic an inflammatory environment. We found that astrocytes with increased FUS levels were more sensitive to IL1β, as shown by their enhanced expression of inflammatory genes, compared with control astrocytes. Moreover, astrocytes overexpressing FUS promoted neuronal cell death and pro-inflammatory microglia activation. We conclude that overexpression of wild-type FUS intrinsically affects astrocyte reactivity and drives their properties toward pro-inflammatory and neurotoxic functions, suggesting that a non-cell autonomous mechanism can support neurodegeneration in FUS-mutated animals and patients.
Detection and characterisation of canine astrovirus, canine parvovirus and canine papillomavirus in puppies using next generation sequencing
Authors: Bhatta Tarka Raj , Chamings Anthony , Vibin Jessy , Alexandersen Soren
Published: March 2019

Gastroenteritis in young animals is a clinical presentation with many infectious and non- infectious aetiologies. We used next generation sequencing (NGS) to investigate the possible infectious causes of gastroenteritis in puppies from a dog kennel in Victoria, Australia. The near complete genome of a canine astrovirus was obtained from pooled faecal samples, and was found to be 94.7% identical with a canine astrovirus detected in the United Kingdom in 2012. The phylogenetic analysis of the capsid gene found similarities to those of canine astroviruses identified in Italy in 2005 and in UK and Hungary in 2012, but distant from that of a canine astrovirus previously identified in Australia in 2012. Thus, different serotypes of canine astrovirus are likely circulating in Australia. The close relationship to European astroviruses also suggested that there had been recent movements of ancestor canine astroviruses between Australia and Europe. NGS also detected other infections in the puppies including several canine papillomaviruses and a canine parvovirus (vaccine strain) as well as a very low level of campylobacter. Canine astrovirus was the probable cause of diarrhoea in these puppies, with the possible involvement of campylobacter bacteria. NGS was effective as a non-targeted method to determine the likely infectious cause of gastroenteritis.
Association of symptoms of attention deficit-hyperactivity disorder and impulsive-aggression with severity of suicidal behavior in adult attempters
Authors: Conejero I. , Jaussent I. , Lopez R. , Guillaume S. , Olié E. , Hebbache C. , Cohen R. F. , Kahn J. P. , Leboyer M. , Courtet P. , Lopez-Castroman J.
Published: March 2019

Literature emphasizes the relationship between attention deficit-hyperactivity disorder (ADHD) and suicidal behavior (SB). However, the link between ADHD and the severity of SB is yet to be determined. We investigated the association between a probable diagnosis of ADHD and the severity of SB in 539 hospitalized suicide attempters, and determined the role of comorbid psychiatric diagnoses. The severity of SB was defined as the number of suicide attempts, age at first suicide attempt, seriousness and violence of suicide attempts. A diagnosis of probable adult ADHD (probable ADHD) was defined as the presence of both current ADHD symptoms and ADHD symptoms in childhood. We evaluated the combined effect of high impulsive-aggression levels and probable ADHD. Probable ADHD was not associated with early or frequent suicide attempts after adjustment for psychiatric disorders and treatment intake. High levels of impulsive-aggression increased the risk of an early suicide attempt, particularly in patients with ADHD symptoms, and independently of other clinical factors. The association between serious suicide attempts and probable ADHD remained significant after adjustment. Although ADHD is involved in suicidal vulnerability, psychiatric comorbidities and impulsive-aggression appear to largely explain the severity of SB in adult attempters with ADHD symptoms.
Steroidogenic Factor 1 (Nr5a1) is Required for Sertoli Cell Survival Post Sex Determination
Authors: Anamthathmakula Prashanth , Miryala Chandra Suma Johnson , Moreci Rebecca S. , Kyathanahalli Chandrashekara , Hassan Sonia S. , Condon Jennifer C. , Jeyasuria Pancharatnam
Published: March 2019

The elevated level of Steroidogenic Factor 1 (Nr5a1, Sf-1) expression in the male gonadal development pathway, post sex determination, implies a vital role in testis gonadal differentiation. In this study we generated Sertoli cell-specific Nr5a1 KO mice (SC-SF-1−/−) at E14.5, which coincides with testis development post sex determination, using the Amh-Cre mouse model. Analysis of SC-SF-1−/− (Sertoli cell specific Nr5a1 knockout) testes demonstrated apoptosis as early as E15. Further analysis revealed that SC-SF-1−/− gonads displayed lower MDM2 levels resulting in elevated TP53 levels, which we believe may lead to apoptosis of the Sertoli cell population, inferring the possibility that NR5A1 directly regulates MDM2 expression. By E15.5, the Sertoli cell and germ cell population declined in SC-SF-1−/− mice resulting in the disruption of seminiferous cords with limited cord structure remaining at E18.5. Due to the loss of Sertoli and germ cells, the testis weights of SC-SF-1−/− mice at 6-weeks were much reduced; however, SC-SF-1−/− seminal vesicles weights were comparable suggesting intact Leydig cell androgen production. We conclude that NR5A1 regulates the TP53 pathway during development, is essential for fetal Sertoli cell survival and controls the cell cycle of Sertoli cells during differentiation.
Assessment of arterial stiffness variables in patients with rheumatoid arthritis: A mediation analysis
Authors: Taverner Delia , Paredes Silvia , Ferré Raimon , Masana Lluís , Castro Antoni , Vallvé Joan-Carles
Published: March 2019

We aimed to study arterial stiffness variables in patients with rheumatoid arthritis (RA), specifically considering their associations with path model mediation analysis. We examined arterial stiffness expressed by the pulse wave velocity (PVW), augmentation index (AIx), distensibility, and clinical and biochemical characteristics in a cohort of 214 RA patients. Variable associations were analysed using multivariate linear regression analysis. We also used path model mediation analysis for PWV variable. Our results indicate that age, systolic blood pressure (SBP), and body mass index (BMI) were significantly associated with PWV, and collectively accounted for 32% of PWV variability. The parallel mediation analysis showed that SBP and BMI accounted for 21% and 7% (a total of 28%) of the total effect of age on PWV, respectively, indicating a partial mediation effect. The associated variables with AIx were age and tender joint count, while those with distensibility were BMI and sex, overall accounting for 16.5% and 4.7% of the variation in AIx and distensibility, respectively. We observed no associations of arterial stiffness with inflammatory variables, disease activity and duration, or cholesterol levels. In conclusion, in our population of RA patients, age is the most important variable that determines the increase in PWV. We have also shown that a significant proportion of the negative effects of age on PWV occurs through increases in SBP and BMI. In our study, lipid and inflammation variables were not associated with an increase in arterial stiffness.
Tracking Decitabine Incorporation into Malignant Myeloid Cell DNA in vitro and in vivo by LC-MS/MS with Enzymatic Digestion
Authors: Chilakala Sujatha , Feng Ye , Li Lan , Mahfouz Reda , Quteba Ebrahem , Saunthararajah Yogen , Xu Yan
Published: March 2019

The DNA hypomethylating agents decitabine and 5-azacytidine are the only two drugs approved for treatment of all subtypes of the myeloid malignancy myelodysplastic syndromes (MDS). The key to drug activity is incorporation into target cell DNA, however, a practical method to measure this incorporation is un-available. Here, we report a sensitive and specific LC-MS/MS method to simultaneously measure decitabine incorporation and DNA hypomethylation. A stable heavy isotope of 2′-deoxycytidine was used as an internal standard and one-step multi-enzyme digestion was used to release the DNA bound drug. Enzyme-released decitabine along with other mononucleosides were separated by a reverse-phase C18column and quantified by mass spectrometry using multiple-reaction-monitoring (MRM) mode, with a lower limit of quantitation at 1.00 nM.In vitrostudies demonstrated dosage and time-dependent incorporation of decitabine into myeloid leukemia cell DNA that correlated with extent of DNA hypomethylation. When applied to clinical samples serially collected from MDS patients treated with decitabine, the method again demonstrated correlation between decitabine DNA-incorporation and DNA hypomethylation. This novel assay to measure the intended molecular pharmacodynamic effect of decitabine therapy can therefore potentially provide insights into mechanisms underlying sensitivity versus resistance to therapy.
Serum lipidomic analysis from mixed neutron/X-ray radiation fields reveals a hyperlipidemic and pro-inflammatory phenotype
Authors: Laiakis Evagelia C. , Canadell Monica Pujol , Grilj Veljko , Harken Andrew D. , Garty Guy Y. , Astarita Giuseppe , Brenner David J. , Smilenov Lubomir , Fornace Albert J.
Published: March 2019

Heightened threats for nuclear terrorism using improvised nuclear devices (IND) necessitate the development of biodosimetry assays that could rapidly assess thousands of individuals. However, the radiation exposures from an IND may be complex due to mixed fields of neutrons and photons (γ-rays), shielding from buildings, and proximity to the epicenter among others. In this study we utilized lipidomics to analyze serum samples from mice exposed to various percentages of neutrons and X-rays to a total dose of 3 Gy. Triacylglycerides, phosphatidylserines, lysophosphatidylethanolamines, lysophosphatidylcholines (LPCs), sphingolipids, and cholesteryl esters all showed delayed increases at day 7 compared to day 1 after irradiation, while diacylglycerides decreased in mixed field exposures and phosphatidylcholines (PCs) remained largely unchanged. Individual lipid molecules with a high degree of unsaturation exhibited the highest fold changes in mixed fields compared to photons alone. More importantly, the increased ratio of LPCs to PCs of each irradiation group compared to control could be used as a radiation biomarker and highlights the existence of a pro-inflammatory phenotype. The results showed that even a small percentage of neutrons in a mixed field can lead to high biological responses with implications for accurate biodosimetry, triage and medical managements of exposed populations.
Pharmacokinetic and pharmacodynamic actions of clozapine-N-oxide, clozapine, and compound 21 in DREADD-based chemogenetics in mice
Authors: Jendryka Martin , Palchaudhuri Monika , Ursu Daniel , van der Veen Bastiaan , Liss Birgit , Kätzel Dennis , Nissen Wiebke , Pekcec Anton
Published: March 2019

Muscarinic Designer Receptors Exclusively Activated by Designer Drugs (DREADD) gated by clozapine-N-oxide (CNO) allow selective G-protein cascade activation in genetically specified cell-types in vivo. Here we compare the pharmacokinetics, off-target effects and efficacy of CNO, clozapine (CLZ) and compound 21 (Cmpd-21) at the inhibitory DREADD human Gi-coupled M4 muscarinic receptor (hM4Di). The half maximal effective concentration (EC50) of CLZ was substantially lower (0.42 nM) than CNO (8.1 nM); Cmpd-21 was intermediate (2.95 nM). CNO was back-converted to CLZ in mice, and CLZ accumulated in brain tissue. However, CNO itself also entered the brain, and free cerebrospinal fluid (CSF) levels were within the range to activate hM4Di directly, while free (CSF) CLZ levels remained below the detection limit. Furthermore, directly injected CLZ was strongly converted to its pharmacologically active metabolite, norclozapine. Cmpd-21 showed a superior brain penetration and long-lasting presence. Although we identified a wide range of CNO and Cmpd-21 off-targets, there was hardly any nonspecific behavioural effects among the parameters assessed by the 5-choice-serial-reaction-time task. Our results suggest that CNO (3–5 mg/kg) and Cmpd-21 (0.4–1 mg/kg) are suitable DREADD agonists, effective at latest 15 min after intraperitoneal application, but both require between-subject controls for unspecific effects.