bluuebunny/crossref_metadata_2025 huggingface dataset:
Functional role of respiratory supercomplexes in mice: segmentation of the Qpool and SCAF1
Authors: Calvo Enrique , Cogliati Sara , Hernansanz-Agustín Pablo , Loureiro-López Marta , Guarás Adela , Casuso Rafael A. , García-Marqués Fernando , Acín-Pérez Rebeca , Martí-Mateos Yolanda , Silla-Castro JC. , Carro-Alvarellos Marta , Huertas Jesús R. , Vázquez Jesús , Enríquez J.A.
Published: November 2019

Mitochondrial respiratory complexes assemble into different forms of supercomplexes (SC). In particular, SC III2+IV require the SCAF1 protein. However, the structural role of this factor in the formation of the respirasome (I+III2+IV) and the physiological role of SCs are controversial. Here, we study C57BL/6J mice harbouring either non-functional SCAF1, the full knock-out for SCAF1 or the wild-type version of the protein and found a growth and exercise phenotype due to the lack of functional SCAF1. By combining quantitative data-independent proteomics, high resolution 2D Blue Native Gel Electrophoresis and functional analysis of enriched respirasome fractions, we show that SCAF1 confers structural attachment between III2 and IV within the respirasome, increases NADH-dependent respiration and reduces ROS production. Furthermore, through the expression of AOX in cells and mice we confirm that CI-CIII superassembly segments the CoQ in two pools and modulates CI-NADH oxidative capacity. These data demonstrate that SC assembly, regulated by SCAF1, modulates the functionality of the electron transport chain.
Experimental control of macrophage pro-inflammatory dynamics using predictive models
Authors: Weinstock Laura D. , Forsmo James E. , Wilkinson Alexis , Ueda Jun , Wood Levi B.
Published: November 2019

Macrophage activity is a major component of the healthy response to infection and injury that consists of tightly regulated early pro-inflammatory activation followed by anti-inflammatory and regenerative activity. In numerous diseases, however, macrophage polarization becomes dysregulated and can not only impair recovery, but can also promote further injury and pathogenesis, e.g. after injury, in diabetes, or in Alzheimer’s disease. Dysregulated macrophages may either fail to polarize or become chronically polarized, resulting in increased production of cytotoxic factors, diminished capacity to clear pathogens, or failure to promote tissue regeneration. In these cases, a method of predicting and dynamically controlling macrophage polarization will enable a new strategy for treating diverse inflammatory diseases. In this work, we developed a model-predictive control framework to temporally regulate macrophage polarization. Using RAW 264.7 macrophages as a model system, we enabled temporal control by identifying transfer function models relating the polarization marker iNOS to exogenous pro- and anti-inflammatory stimuli. These stimuli-to-iNOS response models were identified using linear autoregressive with exogenous input terms (ARX) equations and were coupled with nonlinear elements to account for experimentally identified supra-additive and hysteretic effects. Using this model architecture, we were able to reproduce experimentally observed temporal iNOS dynamics induced with lipopolysaccharides (LPS) and interferon gamma (IFN-γ). Moreover, the identified model enabled the design of time-varying input trajectories to experimentally sustain the duration and magnitude of iNOS expression. By designing transfer function models with the intent to predict cell behavior, we were able to predict and experimentally obtain temporal regulation of iNOS expression using LPS and IFN-γ from both naïve and non-naïve initial states. Moreover, our data driven models revealed decaying magnitude of iNOS response to LPS stimulation over time that could be recovered using combined treatment with both LPS and IFN-γ. Given the importance of dynamic tissue macrophage polarization and overall inflammatory regulation to a broad number of diseases, the temporal control methodology presented here will have numerous applications for regulating immune activity dynamics in chronic inflammatory diseases.
A statistical framework for measuring reproducibility and replicability of high-throughput experiments from multiple sources
Authors: Ranalli Monia , Lyu Yafei , Koch Hillary , Li Qunhua
Published: November 2019

Summary

Replication is essential to reliable and consistent scientific discovery in high-throughput experiments. Quantifying the replicability of scientific discoveries and identifying sources of irreproducibility have become important tasks for quality control and data integration. In this work we introduce a novel statistical model to measure the reproducibility and replicability of findings from replicate experiments in multi-source studies. Using a nested copula mixture model that characterizes the interdependence between replication experiments both across and within sources, our method quantifies reproducibility and replicability of each candidate simultaneously in a coherent framework. Through simulation studies, an ENCODE ChIP-seq dataset and a SEQC RNA-seq dataset, we demonstrate the effectiveness of our method in diagnosing the source of discordance and improving the reliability of scientific discoveries.
Capturing the Continuous Complexity of Behavior inC. elegans
Authors: Ahamed Tosif , Costa Antonio C. , Stephens Greg J.
Published: November 2019

Animal behavior is often quantified through subjective, incomplete variables that may mask essential dynamics. Here, we develop a behavioral state space in which the full instantaneous state is smoothly unfolded as a combination of short-time posture dynamics. Our technique is tailored to multivariate observations and extends previous reconstructions through the use of maximal prediction. Applied to high-resolution video recordings of the roundwormC. elegans, we discover a low-dimensional state space dominated by three sets of cyclic trajectories corresponding to the worm’s basic stereotyped motifs: forward, backward, and turning locomotion. In contrast to this broad stereotypy, we find variability in the presence of locally-unstable dynamics, and this unpredictability shows signatures of deterministic chaos: a collection of unstable periodic orbits together with a positive maximal Lyapunov exponent. The full Lyapunov spectrum is symmetric with positive, chaotic exponents driving variability balanced by negative, dissipative exponents driving stereotypy. The symmetry is indicative of damped, driven Hamiltonian dynamics underlying the worm’s movement control.
Soil Water Carrying Capacity for Vegetation
Authors: Guo Zhong-sheng
Published: November 2019

Non-native plantation can effectively conserve soil and water and improve ecological environment,but soil desiccation occurs and then changes to severe desiccation in most forest and grass lands of water-limited regions, which causes soil degradation, vegetation decline and crop failure, so it is necessary to regulate the changed relationship between plant growth and soil water. However, there is a lack of a universally accepted theory to provide guidance of sustainable use of soil water resources and sustainable management of forest and sustainable produce of crop in such regions. The theory of regulating the relationship between plant growth and soil water in water-limited regions includes the Soil Water Resources Use Limit by Plant (SWRULP) and Soil Water Carrying Capacity for Vegetation (SWCCV). The SWCCV is the population in a plant population or density of indicator plants in a plant community when the soil water supply is equal to soil water consumption in the root zone and a given time, and changes with plant community type, location and time. The degree of coverage, productivity and benefits of a plant community when population quantity of an indicator plant equals SWCCV should be the theoretical basis for ensuring sustainable use of soil water resources and sustainable development of forest.
Maternal Odor Reduces the Neural Response to Fearful Faces in Human Infants
Authors: Jessen Sarah
Published: November 2019

Abstract

Maternal odor is known to play an important role in mother-infant-interaction in many altricial species such as rodents. However, we only know very little about its role in early human development. The present study therefore investigated the impact of maternal odor on infant brain responses to emotional expression. We recorded the electroencephalographic (EEG) signal of seven-month-old infants watching happy and fearful faces. Infants in two control groups exposed to no specific odor (control 1) or the odor of a different infant’s mother (control 2) showed the expected EEG fear response. Crucially, this response was markedly absent in the experimental group exposed to their mother’s odor. Thus, infants respond differently to fear signals in the presence of maternal odor. Our data therefore suggest that maternal odor can be a strong modulator of social perception in human infants.
Nutrient sensing; transcriptomic response and regulation of gut motility in an agastric vertebrate
Authors: Le Hoang T. M. D. , Lie Kai K. , Etayo Angela , Rønnestad Ivar , Sæle Øystein
Published: November 2019

The transcriptome of nutrient sensing and the regulation of gut motility by nutrients in a stomachless fish with a short digestive tract; the ballan wrasse (Labrus berggylta) were investigated. Using an in vitro model, we differentiate how signals initiated by physical stretch and nutrients modulate the gut evacuation rate and motility patterns, and transcriptomic changes. Stretch on the intestine by inert cellulose initiated fast evacuation out of the anterior intestine compared to the digestible protein and lipid. Stretch on the intestine upregulated genes associated with increased muscle activity, whereas nutrients stimulated pathways related to ribosomal activity and the increase in the expression of several neuropeptides which are directly involved in gut motility regulation. Our findings show that physical pressure in the intestine initiate contractions propelling the matter towards the exit, whereas the sensing of nutrients modulates the motility to prolong the residence of digesta in the digestive tract for optimal digestion.

Summary statement

Pressure by food speed up peristalsis in the intestine, but the intestines ability to sense nutrients slow down peristalsis for better digestion. This is partly controlled by genetic regulation.
A model to study orienting responses in zebrafish, and applications towards the emotion-cognition interaction
Authors: do Nascimento Bianca Gomes , Oliveira e Oliveira Hingrid Suzzan Tarso , Lima Silva Hadda Tercya , de Siqueira-Silva Diógenes Henrique , Lima-Maximino Monica , Maximino Caio
Published: November 2019

Orienting responses (ORs) are whole-organism reflexes that are elicited by innocuous stimuli, and which decrease in magnitude after stimulus repetition. ORs represent relatively simple responses that can be used to study attentional processes, and are modulated by the organism’s state, including arousal and activation levels, as well as by emotional processes. Here we describe a simple method to study ORs in zebrafish, a model organism increasingly being used in behavioural neuroscience. After presentation of a static visual stimulus, an OR is elicited, characterized by approaching the stimulus and orienting towards it. After repeated stimulation, OR decreases, suggesting habituation. These responses are qualitatively altered by exposure to a fear-eliciting alarm substance (i.e., derived from the skin of a conspecific), since exposed animals avoid the visual stimulus and orient either away from the stimulus or towards it, but at a distance. The protocol can be used to study orienting responses, as well as the impact of fear and arousal on these reflexes.
Kinetic Analysis of Lipid Metabolism in Live Breast Cancer Cells via Nonlinear Optical Microscopy
Authors: Hou J. , Reid N. E. , Tromberg B. J. , Potma E. O.
Published: November 2019

Investigating the behavior of breast cancer cells via reaction kinetics may help unravel the mechanisms that underlie metabolic changes in tumors. However, obtaining human in vivo kinetic data is challenging due to difficulties associated with measuring these parameters. Non-destructive methods of measuring lipid content in live cells, provide a novel approach to quantitatively model lipid synthesis and consumption. In this study, two-photon excited fluorescence (TPEF) was used to determine metabolic rates via the cell’s optical redox ratio (ORR) as reported by fluorescence intensity ratios of metabolic coenzymes, nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD+). Concurrently, coherent Raman scattering (CRS) microscopy was used to probe de novo intracellular lipid content. Combining non-linear optical microscopy and Michaelis-Menten-kinetics based simulations, we isolated fatty acid synthesis/consumption rates and elucidated effects of altered lipid metabolism in T47D breast cancer cells. When treated with 17β-Estradiol (E2), cancer cells showed a 3-fold increase in beta-oxidation rate as well as a 50% increase in cell proliferation rate. Similarly, the rate of de novo lipid synthesis in cancer cells treated with E2 was increased by 60%. Furthermore, we treated T47D cells with etomoxir (ETO) and observed that cancer cells treated with ETO exhibited a ∼70% reduction in β-oxidation. These results show the ability to probe lipid alterations in live cells with minimum interruption, to characterize both glucose and lipid metabolism in breast cancer cells via quantitative kinetic models and parameters.

Statement of Significance

Combining non-linear optical microscopy (NLOM) and deuterium labeling provides insight into lipid metabolism in live cancer cells during cancer development and progression. The dynamic metabolic data is modelled with Michaelis-Menten-kinetics to independently quantify the lipid synthesis and utilization in cancer cells. Changes in lipid levels are found to originate from de novo lipid synthesis using glucose as a source, lipid consumption from β-oxidation and lipid consumption from cell proliferation, processes that can separately analyzed with the Michaelis-Menten model. In this work, we isolate fatty acid synthesis/consumption rates and elucidated effects of altered lipid metabolism in T47D breast cancer cells in response to estradiol stimulation and etomoxir treatment, dynamic processes that cannot be easily observed without the application of appropriate models.
Assessment of anesthesia on physiological stability and bold signal reliability during visual or acoustic stimulation in the cat
Authors: Levine Alexandra , Li Benson , Barnes Paisley , Lomber Stephen G. , Butler Blake E.
Published: November 2019

Background

Neuroimaging methods including fMRI provide powerful tools to observe whole-brain functional networks. This is particularly powerful in animal models, allowing these networks to be probed using complementary methods. However, most animals must be anesthetized for neuroimaging, giving rise to complications resulting from anesthetic effects on the animal’s physiological and neurological functions. For example, an established protocol for feline neuroimaging involves co-administration of ketamine and isoflurane – the latter of which is known to suppress cortical function.

New Method

Here, we compare this established protocol to alfaxalone, a single-agent anesthetic for functional neuroimaging. We first compare the two in a controlled environment to assess relative safety and to measure physiological stability over an extended time window. We then compare patterns of auditory and visually-evoked activity measured at 7T to assess mean signal strength and between-subjects signal variability.

Results in Comparison with Existing Methods

We show that alfaxalone results in more stable respiratory rates over the 120 minutes testing period, with evidence of smaller between measurements variability within this time window, when compared to ketamine plus isoflurane. Moreover, we demonstrate that both agents evoke similar mean BOLD signals across animals, but that alfaxalone elicits more consistent BOLD activity in response to sound stimuli across all ROIs observed.

Conclusions

Alfaxalone is observed to be more physiologically stable, evoking a more consistent BOLD signal across animals than the co-administration of ketamine and isoflurane. Thus, an alfaxalone-based protocol may represent a better approach for neuroimaging in animal models requiring anesthesia.
A Graph-Directed Approach for Creation of a Homology Modeling Library: Application to Venom Structure Prediction
Authors: Mansbach Rachael A , Chakraborty Srirupa , Travers Timothy , Gnanakaran S.
Published: November 2019

Many toxins are short, cysteine-rich peptides that are of great interest as novel therapeutic leads and of great concern as lethal biological agents due to their high affinity and specificity for various receptors involved in neuromuscular transmission. To perform initial candidate identification for design of a drug impacting a particular receptor or for threat assessment as a harmful toxin, one requires a set of candidate structures of reasonable accuracy with potential for interaction with the target receptor. In this article, we introduce a graph-based algorithm for identifying good extant template structures from a library of evolutionarily-related cysteine-containing sequences for structural determination of target sequences by homology modeling. We employ this approach to study the conotoxins, a set of toxin peptides produced by the family of aquatic cone snails. Currently, of the approximately six thousand known conotoxin sequences, only about three percent have experimentally characterized three-dimensional structures, leading to a serious bottleneck in identifying potential drug candidates. We demonstrate that the conotoxin template library generated by our approach may be employed to perform homology modeling and greatly increase the number of characterized conotoxin structures. We also show how our approach can guide experimental design by identifying and ranking sequences for structural characterization in a similar manner. Overall, we present and validate an approach for venom structure modeling and employ it to expand the library of extant conotoxin structures by almost 300% through homology modeling employing the template library determined in our approach.
The thrombopoietin receptor agonist eltrombopag inhibits human cytomegalovirus replication via iron chelation
Authors: Vogel Jens-Uwe , Schmidt Sophie , Schmidt Daniel , Rothweiler Florian , Koch Benjamin , Baer Patrick C. , Rabenau Holger F. , Michel Detlef , Stamminger Thomas , Michaelis Martin , Cinatl Jindrich
Published: November 2019

The thrombopoietin receptor agonist eltrombopag was successfully used against human cytomegalovirus (HCMV)-associated thrombocytopenia refractory to immunomodulatory and antiviral drugs. These effects were ascribed to effects of eltrombopag on megakaryocytes. Here, we tested whether eltrombopag may also exert direct antiviral effects. Therapeutic eltrombopag concentrations inhibited HCMV replication in human fibroblasts and adult mesenchymal stem cells infected with six different virus strains and drug-resistant clinical isolates. Eltrombopag also synergistically increased the anti-HCMV activity of the mainstay drug ganciclovir. Time-of-addition experiments suggested that eltrombopag interferes with HCMV replication after virus entry. Eltrombopag was effective in thrombopoietin receptor-negative cells, and addition of Fe3+prevented the anti-HCMV effects, indicating that it inhibits HCMV replication via iron chelation. This may be of particular interest for the treatment of cytopenias after haematopoietic stem cell transplantation, as HCMV reactivation is a major reason for transplantation failure. Since therapeutic eltrombopag concentrations are effective against drug-resistant viruses and synergistically increase the effects of ganciclovir, eltrombopag is also a drug repurposing candidate for the treatment of therapy-refractory HCMV disease.
Spermiogenesis in the acoelSymsagittifera roscoffensis: nucleus-plasma membrane contact sites and microtubules
Authors: Hayes Matthew J. , Zakrzewski Anne-C. , Levine Tim P. , Telford Maximilian J.
Published: November 2019

Symsagittifera roscoffensisis a small marine worm found in the intertidal zone of sandy beaches around the European shores of the Atlantic.S. roscoffensisis a member of the Acoelomorpha, a group of flatworms formerly classified with the Platyhelminthes, but now recognised as Xenacoelomorpha, a separate phylum of disputed affinity. We have used electron microscopy to examine the process of spermiogenesis (the final stage of spermatogenesis) inS. roscoffensis, by which spermatids form highly elongated spermatozoa. Their nuclei are long and thread-like, running most of the cell’s length and during the process a pair of flagella are fully incorporated into the cell body. Two previously undescribed inter-organelle contact sites form at different stages of spermiogenesis. Strikingly, there is an extensive nucleus-plasma membrane contact site. Golgi-derived granules containing electron-dense filaments line up along the spermatid plasma membrane, undergo a conformational change, and donate material that forms a peri-nuclear layer that cements this contact site. We also show in earlier stage spermatids that the same granules are associated with microtubules, presumably for traffic along the elongating cell. We identify a second spermiogenesis-specific contact site where sheaths engulfing each internalising flagellum contact the nuclear envelope. Finally, detailed studies of the spermatozoon axonemes show that the central keel has varying numbers of microtubules along the length of the cell, and is likely to be a centriole derivative.

Summary sentence

During spermiogenesis in the acoel flatwormSymsagittifera roscoffensis,two previously unidentified contact sites contribute to the structure of the mature spermatozoon and the axonemal structures show direct continuity between doublet and dense core microtubules.
Impaired Hippocampal-cortical interactions during sleep and memory reactivation without consolidation in a mouse model of Alzheimer’s disease
Authors: Benthem S. D. , Skelin I. , Moseley S. C. , Dixon J. R. , Melilli A. S. , Molina L. , McNaughton B. L. , Wilber A. A.
Published: November 2019

Spatial learning is impaired in preclinical Alzheimer’s disease (AD). We reported similar impairments in 3xTg-AD mice learning aspatial reorientation task. Memory reactivation during sleep is critical for learning related plasticity, and memory consolidation is correlated with hippocampal sharp wave ripple (SWR) density, cortical delta waves (DWs), and their temporal coupling - postulated as a physiological substrate of memory consolidation. Finally, hippocampal-cortical dyscoordination is prevalent in individuals with AD. Thus, we hypothesized impaired memory consolidation mechanisms in hippocampal-cortical networks could account for spatial memory deficits. We assessed sleep architecture, SWR/DW dynamics and memory reactivation in a mouse model of tauopathy and amyloidosis implanted with a recording array targeting isocortex and hippocampus. Mice underwent daily recording sessions of rest-task-rest while learning thespatial reorientation task. We assessed memory reactivation by matching activity patterns from the approach to the unmarked reward zone to patterns during slow wave sleep (SWS). AD mice had more SWS, but reduced SWR density. The increased SWS compensated for reduced SWR density so there was no reduction in SWR number. Conversely, DW density was not reduced so the number of DWs was increased. In control mice hippocampal SWR-cortical DW coupling was strengthened in post-task-sleep and was correlated with performance on thespatial reorientation taskthe following day. However, in AD mice SWR-DW coupling was reduced and not correlated with behavior, suggesting behavioral decoupling. Thus, reduced SWR-DW coupling may cause impaired learning in AD and may serve as a biomarker for early AD related changes.

Significance Statement

Understanding the relationship between network dynamics and cognition early in Alzheimer’s disease progression is critical for identifying therapeutic targets for earlier treatment. We assessed hippocampal-cortical interactions during sleep in AD mice as a potential cause of early spatial learning and memory deficits. We identified compensatory sleep changes in AD mice, that ameliorated some brain dysfunction. Despite the compensatory changes, impaired spatial navigation and impaired hippocampal–cortical (sharp wave ripple-delta wave) interactions were apparent in AD mice. In control but not AD mice hippocampal-cortical interactions were correlated with performance on the spatial task, the following day, suggesting a potential mechanism of impaired consolidation in AD mice. Thus, changes in hippocampal-cortical brain dynamics during sleep may underlie early memory deficits in AD.
Protein function achieved through multiple covalent states
Authors: Butera Diego , Hogg Philip J.
Published: November 2019

The structure of proteins is defined by two main types of covalent bonds; the peptide bonds that link the amino acid residues and disulfide bonds that link pairs of cysteine amino acids. Disulfide bonds are introduced during protein folding and their formation is assumed to be complete in the mature, functional protein. We tested this assumption by quantifying the redox state of disulfide bonds in human blood proteins in their native environment. Using a differential cysteine alkylation and mass spectrometry method, we measured the redox state of disulfide bonds in circulating fibrinogen and von Willebrand factor. There is an extraordinary disulfide lability in the proteins, with 27 bonds in the two proteins ranging from 3 to 50% reduced in healthy human donors. Modelling of the data indicates that the proteins exist in hundreds of different disulfide-bonded states in the circulation. Different covalent states of fibrinogen are associated with different binding activities and their distribution is changed by fluid shear forces and altered in patients with cardiovascular disease, indicating that the different states have different functions and are dynamic. These findings have implications for protein function generally and how proteins are targeted in experimental settings and for therapeutic purposes.
Spatiotemporal patterns of rodent hippocampal field potentials uncover spatial representations
Authors: Cao Liang , Varga Viktor , Chen Zhe S.
Published: November 2019

Spatiotemporal patterns of large-scale spiking and field potentials of the rodent hippocampus encode spatial representations during maze run, immobility and sleep. Here, we showed that multi-site hippocampal field potential amplitude at ultra-high frequency band (FPAuhf) provides not only a fast and reliable reconstruction of the rodent’s position in wake, but also a readout of replay content during sharp wave ripples. This FPAuhf feature may serve as robust real-time decoding strategy from large-scale (up to 100,000 electrodes) recordings in closed-loop experiments. Furthermore, we developed unsupervised learning approaches to extract low-dimensional spatiotemporal FPAuhf features during run and ripple periods, and to infer latent dynamical structures from lower-rank FPAuhf features. We also developed a novel optical flow-based method to identify propagating spatiotemporal LFP patterns from multi-site array recordings, which can be used for decoding application. Finally, we developed a prospective decoding strategy to predict animal’s future decision in goal-directed navigation.
Pregnancy alters innate immune responses to Zika virus infection in the genital tract
Authors: Lesteberg Kelsey E. , Fader Dana S. , Beckham J. David
Published: November 2019

Recent outbreaks of Zika virus (ZIKV) have been associated with birth defects, including microcephaly and neurological impairment. However, the mechanisms which confer increased susceptibility to ZIKV during pregnancy remain unclear. We hypothesized that poor outcomes from ZIKV infection during pregnancy are due in part to pregnancy-induced alteration of innate immune cell frequencies and cytokine expression. To examine the impact of pregnancy on innate immune responses, we inoculated pregnant and non-pregnant female C57BL/6 mice with 5×105FFU of ZIKV intravaginally. Innate immune cell frequencies and cytokine expression were measured by flow cytometry at day 3 post infection. Compared to non-pregnant mice, pregnant mice exhibited higher frequencies of uterine macrophages (CD68+) and tolerogenic dendritic cells (CD11c+ CD103+ and CD11c+ CD11b+). Additionally, ZIKV-infected pregnant mice had lower frequencies of CD45+ IL-12+ and CD11b+ IL-12+ cells in the uterus and spleen. These data show that pregnancy results in an altered innate immune response to ZIKV infection in the genital tract of mice and that pregnancy-associated immune modulation may play an important role in the severity of acute ZIKV infection.

Importance

Pregnant females longer duration that viremia following infection with Zika virus but the mechanism of this is not established. Innate immune cellular responses are important for controlling virus infection and are important for development and maintenance of pregnancy. Thus, the acute immune response to Zika virus during pregnancy may be altered so that the pregnancy can be maintained. To examine this interaction, we utilized a mouse model of Zika virus infection during pregnancy using intravaginal inoculation. We found that following Zika virus infection, pregnant mice exhibited increased expression of tolerant or non-inflammatory dendritic cells. Additionally, we found that pregnant mice have significantly depressed ability to secrete the cytokine IL-12 from innate immune cells in the uterus and the spleen while maintaining MHCII expression. These findings show that pregnancy-induced changes in the innate immune cells are biased towards tolerance and can result in decreased antigen-dependent stimulation of immune responses.
Six cases of perforating pilomatricoma: Anetodermic changes with expression of matrix metalloproteinases
Authors: Watabe Daisuke , Mori Shiho , Akasaka Toshihide , Motegi Sei‐ichiro , Ishikawa Osamu , Amano Hiroo
Published: November 2019

Perforating pilomatricoma (PP) is a rare clinical variant of pilomatricoma presenting as a crusted or ulcerated nodule. Previous reports have suggested that the tumor cells perforate the epidermis through a process of transepithelial elimination. Here, we report six cases of PP and examine the mechanism of transepithelial elimination in PP. Histologically, the dermis above or around the tumor nest exhibited edema, dilated vascular spaces, sparse collagen bundles and absence of elastic fibers, suggesting anetodermic changes in all cases. Immunohistochemistry demonstrated many CD68‐positive macrophages around the tumor nests. Matrix metallopeptidase (MMP)‐9 and MMP‐12 were expressed in the inflammatory cells and tumor cells, and were also present in the epidermis and fibroblasts in all cases. We speculate that in PP anetodermic change caused by MMP and elastases including MMP‐9 and MMP‐12 may precede elimination of the tumor.
Behavioural complementarity among frugivorous birds and lizards can promote plant diversity in island ecosystems
Authors: Morán‐López Teresa , González‐Castro Aarón , Morales Juan Manuel , Nogales Manuel
Published: November 2019

The behavioural complementarity of fruit‐eating animals is thought to exert a key role in plant community assembly. However, a mechanistic understanding of the causal links between the two processes is still lacking.

This study assesses whether complementarity between dispersers in feeding and microhabitat‐use behaviour enhances community‐scale dispersal services, resulting in a more diverse community of seedlings.

We used a Bayesian approach to connect a comprehensive database of seed dispersal effectiveness at a community scale with a transition probability model that accounts for behavioural complementarity. Our model system was the thermosclerophyllous shrubland of the Canary Islands. There, fleshy‐fruited plants rely on two types of frugivores: lizards and birds.

Lizards consumed all plant species and preferentially used open areas, whereas birds foraged for small single‐seeded fruits and dispersed their seeds beneath plants. Through feeding on different sets of plants, they generated a rich seed‐rain community. By diversifying the microhabitat of deposition, more species could find suitable recruitment sites.

Distinct foraging and microhabitat‐use choices led to complementary dispersal services. Lizards ensured that all plant species were present in the seedling community, while birds promoted a more even distribution of them. As a result, diversity in the community of seedlings was enhanced.

Overall, our work underscores that behavioural complementarity promotes diversity in the early‐regenerating plant communities. These enhanced dispersal services rely on the presence of all functional groups. Thus, in communities where frugivores display unique behaviours, preserving a diverse community of dispersers should be a conservation target.

A free Plain Language Summary can be found within the Supporting Information of this article.
Elevated TRIM23 expression predicts cisplatin resistance in lung adenocarcinoma
Authors: Zhang Youwei , Du He , Li Yang , Yuan Yuan , Chen Bi , Sun Sanyuan
Published: November 2019

The tripartite motif containing 23 (TRIM23) gene is a member of the tripartite motif (TRIM) family that participates in many pathophysiological processes. However, the role of TRIM23 in lung adenocarcinoma (LUAD) remains unclear. In the present study, TRIM23 was first screened by next‐generation sequencing between the cisplatin (DDP)‐resistant A549/DDP cell line and the parental A549 cell line, combined with integrated analysis of the Gene Expression Omnibus (GEO) data (E‐GEOD‐43493 and E‐GEOD‐43494). The expression of TRIM23 was then verified to be upregulated in the DDP‐resistant LUAD cells and tissues. The knockdown of TRIM23 expression in A549/DDP cells caused increased apoptosis, decreased IC50 values of DDP, NF‐κB nuclear translocation, inhibition of cell proliferation in vitro and in vivo, inhibition of GLUT1/3 expression, glucose uptake, and lactate and ATP production. TRIM23 overexpression resulted in the opposite effects in A549 cells. In addition, the inhibition of proliferation in A549 cells caused by NF‐κB signaling inhibitor PTDC or glycolysis inhibitor 3‐BrPA could be weakened by TRIM23 overexpression. Furthermore, immunohistochemical analysis revealed that TRIM23 was upregulated in 46.1% (70/152) of LUAD cases, and elevated TRIM23 expression was correlated with high expression of NF‐κB, poor cellular differentiation, and adverse overall survival (OS) and disease‐free survival (DFS). In conclusion, our study demonstrates that TRIM23 acts as an oncogene in LUAD and promotes DDP resistance by regulating glucose metabolism via the TRIM23/NF‐κB/ GLUT1/3 axis.
Effect of clomiphene citrate treatment on the Sertoli cells of dysmetabolic obese men with low testosterone levels
Authors: Pelusi Carla , Fanelli Flaminia , Baccini Margherita , Triggiani Vincenzo , Bartolomeo Nicola , Carbone Matteo Domenico , De Pergola Giovanni , Di Dalmazi Guido , Pagotto Uberto , Pasquali Renato , Giagulli Vito Angelo
Published: November 2019

Background

Clomiphene citrate (CC) has been shown to restore the hypothalamic‐pituitary‐gonadal (HPG) axis by increasing testosterone (T) levels to physiological levels in patients with dysmetabolic conditions such as obesity, metabolic syndrome and type 2 diabetes mellitus (T2DM). However, the data are unclear regarding the effects on Sertoli cell (SC) function.

Aim

To study SC function by assessing Inhibin B (IB) and anti‐Mullerian hormone (AMH) levels at baseline and after 3 months of CC treatment.

Materials and methods

This is an ancillary study of a cross‐over, randomised, double‐blind, placebo‐controlled trial performed to evaluate androgen response to CC treatment in dysmetabolic obese subjects with low T levels treated with metformin. We evaluated SC function by assessing IB and AMH levels at baseline and after 3 months of each treatment in ten dysmetabolic obese subjects with low T levels. In all subjects, the influence of the clinical characteristics, metabolic and hormonal baseline parameters on SC and Leydig (LC) function, evaluated respectively with AMH, IB, follicle‐stimulating hormone (FSH) and T levels, was tested.

Results

No significant changes were observed for IB and AMH concentrations after each treatment period. Whereas T and oestradiol (E2) levels were shown to be significantly higher in the CC plus metformin phase (CC/Met) only. No clinical, metabolic or hormonal parameters showed significant effects on serum AMH at baseline or after treatments. However, baseline T, dihydrotestosterone (DHT) and E2 positively affected IB levels during CC/Met therapy (P = .003, P = .038 and P = .049, respectively). Baseline leptin and FSH had a negative (P = 031) and positive (P = .048) respectively role on T levels during CC/Met, as they were statistically significant compared to the placebo period (Plac/Met).

Conclusion

Unlike the LC activity, CC was unable to influence SC function, as shown by the lack of IB and AMH serum modifications, thus suggesting an intrinsic nonreversible defect of SC cells in patients with dysmetabolic conditions.
Reduced perforin release from T cells as a mechanism underlying hypothermia‐mediated neuroprotection
Authors: Matsui Tomohiro , Yoshida Yusuke
Published: November 2019

Objective

The underlying mechanisms of therapeutic hypothermia, which protects neurons after severe brain damage, are partially understood. T cells infiltrate the brain within days after cerebral ischemia and play essential roles in exacerbating the delayed phase of brain injury by producing cytotoxic factors, which were also systematically shown to be involved in brain damage. Periphery brain abnormalities are interesting, because the periphery might constitute a pathway to the central nervous system and therefore could also be a target of therapeutic hypothermia. We elucidated the effects of hypothermia and hyperthermia on peripheral T cell‐derived perforin (Pfn), and the underlying mechanism of Pfn‐mediated neurotoxicity and death of brain microvascular endothelial cells (BMVECs).

Methods

We determined the levels of Pfn produced by activated CD4+ and CD8+ T cells obtained from healthy humans under hypothermic, normothermic and hyperthermic conditions. The viability in response to Pfn treatment was assessed in neuronal Neuro‐2a and brain microvascular endothelial bEnd.3 cells.

Results

Compared with normothermia, Pfn release in both CD4+ and CD8+ T cells was reduced by hypothermia, but augmented by hyperthermia. Pfn caused the death of Neuro‐2a cells, and induced both apoptosis and necrosis in bEnd.3 cells; both effects were concentration‐dependent.

Conclusions

Hypothermia reduces T cell‐derived release of Pfn, which mediates neuronal cell death and apoptosis/necrosis of BMVECs, suggesting that therapeutic hypothermia elicits protection in the neurons and BMVECs in the delayed phase of brain injury by directly reducing the release of T cell‐derived Pfn. BMVECs protection can aid in reduced vascular permeability and blood–brain barrier protection.
Performance of bone‐level implants with conical connections in the anterior maxilla: A 5‐year prospective cohort study
Authors: Meijndert Caroliene M. , Raghoebar Gerry M. , Santing Hendrik J. , Vissink Arjan , Meijer Henny J. A.
Published: November 2019

Objectives

To assess clinical, radiographic and aesthetic outcomes, and satisfaction of patients treated with single implant restorations in the maxillary aesthetic region 5 years after final restoration.

Material and methods

Sixty patients (29 male/31 female, mean age 37 years) with a missing anterior tooth in the maxilla (39 central incisors/10 lateral incisors/5 cuspids/6 first premolars) received a bone‐level implant with conical connection. In 29 patients, a bone augmentation procedure was necessary before implant placement (autogenous bone grafts mixed with spongiosa granules). All implants (12 with 3.3 mm diameter/ 48 with 4.1 mm diameter) were loaded after 3 months of submerged healing. The restoration consisted of an individually designed full‐zirconia abutment veneered with porcelain. Follow‐up with clinical and radiographic assessment was conducted until 60 months after the final restoration. Aesthetic outcome of the restoration was determined with the Pink Esthetic Score–White Esthetic Score (PES‐WES). Patient satisfaction was assessed with a VAS scale and satisfaction questionnaire.

Results

Fifty patients completed the 5‐year follow‐up. Implant survival was 100%, restoration survival 98%. Mean bone‐level change was −0.13 ± 0.66 mm with a median (IQR) pocket probing depth of 2.75 [2.25; 3.25]. The mean PES and WES scores were 6.6 ± 1.7 and 7.8 ± 1.5, respectively. Patient satisfaction was high (92.1 ± 7.8 on 100 mm VAS scale). There were no differences between patients with or without a bone augmentation procedure.

Conclusion

Bone‐level implants with a conical connection are a reliable treatment option in single‐tooth replacements in the maxillary aesthetic zone.
Prognosis of oral squamous cell carcinoma with perineural invasion: A comparative study of classification types
Authors: Park Jae , Megow Anna , Swalling Adam , Hodge John‐Charles , Foreman Andrew , Boase Samuel , Valentine Rowan , Krishnan Suren , Ooi Eng H.
Published: November 2019

Objective

To investigate the histological location and extent of perineural invasion (PNI) as prognostic factors.

Design

Retrospective review of medical records and histological analysis of 116 patients with oral squamous cell carcinoma (OSCC).

Setting

Two major public tertiary hospitals treating head and neck cancer, Royal Adelaide Hospital and Flinders Medical Centre, in South Australia.

Participants

Patients diagnosed with OSCC who underwent primary surgical treatment with curative intent at these two centres from January 1, 2005 through December 31, 2015.

Main Outcome Measures

The primary end points were disease‐free survival (DFS) and disease‐specific survival (DSS).

Results

The presence of PNI as a binary factor alone did not significantly influence the clinical outcomes. Extratumoural (ET) PNI as measured from the tumour edge was associated with worse DFS on multivariate analyses. Multifocal PNI was associated with worse DFS and DSS. DFS in multifocal PNI was worse irrespective of whether adjuvant therapy was administered.

Conclusions

The presence of multifocal and ET PNI in OSCC is associated with poor clinical outcomes. Patients with multifocal PNI were associated with worse DFS even with adjuvant therapy.
Matrix metalloproteinases MMP‐1, MMP‐2, and MMP‐13 are overexpressed in primary nodular melanoma
Authors: Zamolo Gordana , Grahovac Maja , Žauhar Gordana , Vučinić Damir , Kovač Leo , Brajenić Nika , Grahovac Blaženka
Published: November 2019

Background

The spread and invasion of malignant melanoma cells involve degradation and reorganization of the extracellular matrix by the activation of several matrix metalloproteinases (MMPs). This study analyzed the expression of MMP‐1, MMP‐2, and MMP‐13 proteins in primary nodular melanoma (NM) and dysplastic nevi (DN) as a significant risk factor for melanoma development. The secondary goal was to analyze the correlation of MMPs protein expression in NM with tumor invasion, BRAF V600 mutation status, and overall survival.

Methods

Immunohistochemistry for MMP‐1, MMP‐2, and MMP‐13 was performed on nodular melanoma (n = 52) and dysplastic nevi (n = 28) on tissue microarray (TMA). BRAF V600 mutation analysis on NM samples was performed by the Sanger sequencing method.

Results

A high level of MMPs expression in NM samples (>30%) compared with DN (<8%) was statistically significant (P < 0.001). BRAF V600 mutations were detected in 15 of 39 (38.5%) NM samples. This study revealed an interesting finding that MMP‐1 and MMP‐13 protein expression in the BRAF V600 mutated melanomas were significantly lower than in the BRAF V600 wild type (P < 0.05).

Conclusion

Cox analysis revealed that Clark categories, Breslow thickness, and MMP‐1 high protein expression are predictive factors for shorter overall survival (P < 0.05).
Intercropping impacts the host location behaviour and population growth of aphids
Authors: Mansion‐Vaquié Agathe , Ferrer Aurélie , Ramon‐Portugal Felipe , Wezel Alexander , Magro Alexandra
Published: November 2019

Increasing intrafield plant diversity has been shown to regulate pest populations in various agroecosystems. Among the suggested mechanisms for this bottom‐up pest control, the disruptive crop hypothesis states that herbivores' abilities to locate and colonize their host plants are reduced by the presence of non‐host plants. Under laboratory conditions, we evaluated how intercropping wheat and legumes modifies the behaviour of apterous cereal aphids,Sitobion avenae(Fabricius) (Hemiptera: Aphididae), in terms of host plant location and population growth. We compared two intercropping systems – soft winter wheat,Triticum aestivumL. (Poaceae), associated with winter pea,Pisum sativumL., or with white clover,Trifolium repensL. (both Fabaceae) – and sole stands of soft winter wheat. Aphids needed more time to locate their wheat host plant and then spent less time on wheat when it was intercropped with clover. At the population level, and accounting for host plant biomass, only intercropping wheat with clover significantly reduced aphid densities on wheat, as this was particularly disruptive toS. avenaebehaviour and population growth.Our laboratory study points out that the species used as non‐host plants and their density are important parameters that should be taken into account in field studies on intercropping systems.
Intercropping oilseed rape as a potential relay crop for enhancing the biological control of green peach aphids and aphid‐transmitted virus diseases
Authors: Lai Rongquan , Hu Hanqing , Wu Xiaoting , Bai Jingjing , Gu Gang , Bai Jianbao , Zhou Ting , Lin Tianran , Zhong Xiujin
Published: November 2019

Tobacco viruses transmitted by green peach aphids, Myzus persicae (Sulzer) (Hemiptera: Aphididae), cause severe disease in flue‐cured tobacco, Nicotiana tabacum L. (Solanaceae), in China and throughout the world. Field experiments were conducted in 2016 and 2017 in Longyan City, Fujian Province, China, to determine whether M. persicae and aphid‐transmitted virus diseases are affected by intercropping of oilseed rape, Brassica napus L. (Brassicaceae), in tobacco fields. The results showed that, compared with those in monocultured fields, the densities of M. persicae and winged aphids in intercropped fields significantly decreased in both 2016 and 2017. In particular, the appearance of winged aphids was delayed by ca. 7 days. Moreover, the densities of Aphidius gifuensis Ashmead (Hymenoptera: Aphidiidae), a parasitoid of the aphid, significantly increased in 2016 and 2017. Accordingly, the incidence rates of aphid‐transmitted virus diseases (those caused by the cucumber mosaic virus, potato virus Y, and tobacco etch virus) significantly decreased in the intercropped fields in 2016 and 2017. Tobacco yields and monetary value significantly increased in 2016 (by 10–25 and 14–29%, respectively) and 2017 (by 17–22 and 22–34%, respectively). Consequently, our results suggest that intercropping oilseed rape in tobacco fields is a good approach to regulating and controlling aphids and tobacco mosaic viruses, for example potyvirus, and this intercropping can help control aphid‐transmitted virus diseases in tobacco.