bluuebunny/crossref_metadata_2025 huggingface dataset:
A Wnt-BMP4 signalling axis induces MSX and NOTCH proteins and promotes growth suppression and differentiation in neuroblastoma
Authors: Szemes Marianna , Melegh Zsombor , Bellamy Jacob , Kollareddy Madhu , Catchpoole Daniel , Malik Karim
Published: February 2020

The Wnt and bone morphogenetic protein (BMP) signalling pathways are known to be crucial in the development of neural crest lineages, including the sympathetic nervous system. Surprisingly, their role in paediatric neuroblastoma, the prototypic tumour arising from this lineage, remains relatively uncharacterised. We previously demonstrated that Wnt/β-catenin signalling can have cell-type specific effects on neuroblastoma phenotypes, including growth inhibition and differentiation, and that BMP4 mRNA and protein were induced by Wnt3a/Rspo2. In this study, we characterise the phenotypic effects of BMP4 on neuroblastoma cells, demonstrating convergent induction of MSX homeobox transcription factors by Wnt and BMP4 signalling and BMP4-induced growth suppression and differentiation. Immunohistochemical analysis of BMP4 expression in primary neuroblastomas confirms a striking absence of BMP4 in poorly differentiated tumours, in contrast to high expression in ganglion cells. These results are consistent with a tumour suppressive role for BMP4 in neuroblastoma. RNA sequencing following BMP4 treatment revealed induction of Notch signalling, verified by increases of Notch3 and Hes1 proteins. Together, our data demonstrate for the first time Wnt-BMP-Notch signalling crosstalk associated with growth suppression of neuroblastoma.
Characterization of the motor cortex transcriptome supports microgial-related key events in amyotrophic lateral sclerosis
Authors: Dols-Icardo Oriol , Montal Victor , Sirisi Sònia , López-Pernas Gema , Cervera-Carles Laura , Querol-Vilaseca Marta , Muñoz Laia , Belbin Olivia , Alcolea Daniel , Molina-Porcel Laura , Pegueroles Jordi , Turón-Sans Janina , Blesa Rafael , Lleó Alberto , Fortea Juan , Rojas-García Ricard , Clarimón Jordi
Published: February 2020

Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease characterized by the degeneration of upper and lower motor neurons. A major neuropathological finding in ALS is the coexistence of glial activation and aggregation of the phosphorylated transactive response DNA-binding protein 43-kDa (pTDP43) in the motor cortex at the earliest stages of the disease. Despite this, the transcriptional alterations associated with these pathological changes in this major vulnerable brain region have yet to be fully characterized. Here, we have performed massive RNA sequencing of the motor cortex of ALS (n=11) and healthy controls (HC; n=8). We report extensive RNA expression alterations at gene and isoform levels, characterized by the enrichment of neuroinflammatory and synapse related pathways. The assembly of gene co-expression modules confirmed the involvement of these two principal transcriptomic changes, and showed a strong negative correlation between them. Furthermore, cell-type deconvolution using human single-nucleus RNA sequencing data as reference demonstrated that microglial cells are overrepresented in ALS compared to HC. Importantly, we also show for the first time in the human ALS motor cortex, that microgliosis is mostly driven by the increased proportion of a microglial subpopulation characterized by gene markers overlapping with the recently described disease associated microglia (DAM). Using immunohistochemistry, we further evidenced that this microglial subpopulation is overrepresented in ALS and that variability in pTDP43 aggregation among patients negatively correlates with the proportion of microglial cells. In conclusion, we report that neuroinflammatory changes in ALS motor cortex are dominated by microglia which is concomitant with a reduced expression of postsynaptic transcripts, in which DAM might have a prominent role. Microgliosis therefore represents a promising avenue for therapeutic intervention in ALS.
EXO70D isoforms mediate selective autophagic degradation of Type-A ARR proteins to regulate cytokinin sensitivity
Authors: Acheampong Atiako Kwame , Shanks Carly , Chang Chai-Yi , Schaller G. Eric , Dagdas Yasin , Kieber Joseph J.
Published: February 2020

The phytohormone cytokinin influences many aspects of plant growth and development, several of which also involve the cellular process of autophagy, including leaf senescence, nutrient re-mobilization, and developmental transitions. The Arabidopsis type-A Response Regulators (type-A ARR) are negative regulators of cytokinin signaling that are transcriptionally induced in response to cytokinin. Here, we describe a mechanistic link between cytokinin signaling and autophagy, demonstrating that plants modulate cytokinin sensitivity through autophagic regulation of type-A ARR proteins. Type-A ARR proteins were degraded by autophagy in an AUTOPHAGY-RELATED (ATG)5-dependent manner. EXO70D family members interacted with Type-A ARR proteins, likely in a phosphorylation-dependent manner, and recruited them to autophagosomes via interaction with the core autophagy protein, ATG8. Consistently, loss-of-function exo70D1,2,3 mutants compromised targeting of type-A ARRs to autophagic vesicles, have elevated levels of type-A ARR proteins, and are hyposensitive to cytokinin. Disruption of both type-A ARRs and EXO70D1,2,3 compromised survival in carbon-deficient conditions, suggesting interaction between autophagy and cytokinin responsiveness in response to stress. These results indicate that the EXO70D proteins act as selective autophagy receptors to target type-A ARR cargos for autophagic degradation, demonstrating that cytokinin signaling can be modulated by selective autophagy.
Telomere length heterogeneity in ALT cells is maintained by PML-dependent localization of the BTR complex to telomeres
Authors: Loe Taylor K , Li Julia Su Zhou , Zhang Yuxiang , Azeroglu Benura , Boddy Michael Nicholas , Denchi Eros Lazzerini
Published: February 2020

Telomeres consist of TTAGGG repeats bound by protein complexes that serve to protect the natural end of linear chromosomes. Most cells maintain telomere repeat lengths by utilizing the enzyme telomerase, although there are some cancer cells that use a telomerase-independent mechanism of telomere extension, termed Alternative Lengthening of Telomeres (ALT). Cells that employ ALT are characterized, in part, by the presence of specialized PML nuclear bodies called ALT-associated PML-Bodies (APBs). APBs localize to and cluster telomeric ends together with telomeric and DNA damage factors, which led to the proposal that these bodies act as a platform on which ALT can occur. However, the necessity of APBs and their function in the ALT pathway has remained unclear. Here, we used CRISPR/Cas9 to delete PML and APB components from ALT-positive cells to cleanly define the function of APBs in ALT. We find that PML is required for the ALT mechanism, and that this necessity stems from APBs’ role in localizing the BLM-TOP3A-RMI (BTR) complex to ALT telomere ends. Strikingly, recruitment of the BTR complex to telomeres in a PML-independent manner bypasses the need for PML in the ALT pathway, suggesting that BTR localization to telomeres is sufficient to sustain ALT activity.
Phosphoproteomic analysis of mammalian infectiveTrypanosoma bruceisubjected to heat shock suggests atypical mechanisms for thermotolerance
Authors: Ooi Cher P. , Benz Corinna , Urbaniak Michael D.
Published: February 2020

The symptoms of African sleeping sickness, caused by the parasiteTrypanosoma brucei, can include periods of fever as high as 41 °C which triggers a heat shock response in the parasite. To capture events involved in sensing and responding to heat shock in the mammalian infective form we have conducted a SILAC-based quantitative proteomic and phosphoproteomic analysis ofT. bruceicells treated at 41 °C for 1h. Our analysis identified 193 heat shock responsive phosphorylation sites with an average of 5-fold change in abundance, but only 20 heat shock responsive proteins with average of 1.5-fold change. These data indicate that protein abundance does not rapidly respond (≤1 h) to heat shock, and that the changes observed in phosphorylation site abundance are larger and more widespread. The heat shock responsive phosphorylation sites showed enrichment of RNA binding proteins with putative roles in heat shock response included P-body / stress granules and the eukaryotic translation initiation 4F complex. The ZC3H11-MKT1 complex, which stabilises mRNAs of thermotolerance proteins, appears to represent a key signal integration node in the heat shock response.
A 3D human liver model of nonalcoholic steatohepatitis
Authors: Duriez Marion , Jacquet Agnes , Hoet Lucile , Roche Sandrine , Bock Marie-Dominique , Rocher Corinne , Haussy Gilles , Vigé Xavier , Bocskei Zsolt , Slavnic Tamara , Martin Valérie , Guillemot Jean-Claude , Didier Michel , Kannt Aimo , Orsini Cécile , Mikol Vincent , Le Fèvre Anne-Céline
Published: February 2020

Abstract

We have developed anin vitropreclinical 3D Non-Alcoholic SteatoHepatitis (NASH) model by co-culturing four human primary liver cell types: hepatocytes, stellate, endothelial and Kupffer cells. Cells were embedded in a hydrogel of rat collagen in 96-well plate and a NASH-like environment was induced with a medium containing free fatty acids (FFAs) and tumor necrosis factor α (TNFα). This model was characterized by biochemical, imaging and transcriptomics analysis. On the one hand, we succeed in defining suitable culture conditions to maintain the 3D co-culture up to 10 daysin vitrowith the lowest level of steatosis, and reproducible low levels of inflammation and fibrosis. On the other hand, we induced NASH disease with a custom medium mimicking NASH features (hepatocyte injury, steatosis, inflammation and fibrosis). The 10-day cell viability and cost effectiveness of the model make it suitable for medium throughput drug screening and provide attractive avenues to better understand disease physiology and to identify and characterize new drug targets.

Summary

We developed a 3D human liver model which exhibits many features of non-alcoholic steatohepatitis and that could become a platform for medium throughput drug screening.
Phasing and imputation of single nucleotide polymorphism data of missing parents of bi-parental plant populations
Authors: Gonen Serap , Wimmer Valentin , Gaynor R. Chris , Byrne Ed , Gorjanc Gregor , Hickey John M.
Published: February 2020

This paper presents an extension to a heuristic method for phasing and imputation of genotypes of descendants in bi-parental populations so that it can phase and impute genotypes of parents of bi-parental populations that are fully ungenotyped or partially genotyped. The imputed genotypes of the parent are then used to impute low-density genotyped descendants of the bi-parental population to high-density. The extension works in three steps. First, it identifies whether a parent has no or low-density genotypes available and it identifies all of its relatives that have high-density genotypes. Second, using the high-density information of relatives, it determines whether the parent is homozygous or heterozygous for a given locus. Third, it phases heterozygous positions of the parent by matching haplotypes to its relatives.

We implemented the new algorithm in an extension of the AlphaPlantImptue software and tested its accuracy of imputing missing parent genotypes in simulated bi-parental populations from different scenarios. We also tested the accuracy of imputation of the missing parent’s descendants using the true genotype of the parent and compared this to using the imputed genotypes of the parent. Our results show that across all scenarios, the accuracy of imputation of a parent, measured as the correlation between true and imputed genotypes, was > 0.98 and did not drop below ∼ 0.96. The imputation accuracy of a parent was always higher when it was inbred than when it was outbred and when it had low-density genotypes. Including ancestors of the parent at HD, increasing the number of crosses and the number of high-density descendants all increased the accuracy of imputation. The high imputation accuracy achieved for the parent across all scenarios translated to little or no impact on the accuracy of imputation of its descendants at low-density.

Key Message

New fast and accurate method for phasing and imputation of SNP chip genotypes within diploid bi-parental plant populations.
The use of nanovibration to discover specific and potent bioactive metabolites that stimulate osteogenic differentiation in mesenchymal stem cells
Authors: Hodgkinson Thomas , Tsimbouri P. Monica , Llopis-Hernandez Virginia , Campsie Paul , Scurr David , Childs Peter G , Phillips David , Donnelly Sam , Wells Julia A , Salmeron-Sanchez Manuel , Burgess Karl , Alexander Morgan , Vassalli Massimo , Oreffo Richard O.C. , Reid Stuart , France David J , Dalby Matthew J
Published: February 2020

Bioactive metabolites have wide-ranging biological activities and are a potential source of future research and therapeutic tools. Here, we use nanovibrational stimulation to induce the osteogenic differentiation of MSCs, in the absence of off-target differentiation. We show that this differentiation method, which does not rely on the addition of exogenous growth factors to the culture media, provides an artefact-free approach to identifying bioactive metabolites that specifically and potently induce osteogenesis. We first identify a highly specific metabolite as the endogenous steroid, cholesterol sulphate. Next, a screen of other small molecules with a similar steroid scaffold identified fludrocortisone acetate as being both specific and having highly potent osteogenic-inducing activity. These findings demonstrate that physical priciples can be used to identify bioactive metabolites and then metabolite potency can be optimised by examining structure-function relationship.
SPAs promote thermomorphogenesis via regulating the phyB-PIF4 module inArabidopsis
Authors: Lee Sanghwa , Paik Inyup , Huq Enamul
Published: February 2020

High ambient temperature due to global warming has a profound influence on plant growth and development at all stages of life cycle. Plant response to high ambient temperature termed thermomorphogenesis is characterized by hypocotyl and petiole elongation, and hyponastic growth at seedling stage. However, the molecular mechanism of thermomorphogenesis is still rudimentary. Here, we show that a set of fourSUPPRESSOR OF PHYA-105(SPA) genes are required for thermomorphogenesis. Consistently, SPAs are necessary for global gene expression changes in response to high ambient temperature. SPA1 level is unaffected, while the thermosensor phyB is stabilized in thespaQmutant at high ambient temperature. Furthermore, in the absence of fourSPAgenes, the pivotal transcription factor PIF4 fails to accumulate, indicating a role of SPAs in regulating the phyB-PIF4 module at high ambient temperature. SPA1 directly phosphorylates PIF4in vitro, and a mutant SPA1 affecting the kinase activity fails to rescue the PIF4 level as well as the thermo-insensitive phenotype ofspaQ, suggesting that the SPA1 kinase activity is necessary for thermomorphogenesis. Taken together, these data suggest that SPAs integrate light and temperature signaling via fine tuning the phyB-PIF4 module.
Mitochondria dynamics in postmitotic cells drives neurogenesis through Sirtuin-dependent chromatin remodeling
Authors: Iwata Ryohei , Vanderhaeghen Pierre
Published: February 2020

The conversion of neural stem cells into neurons is associated with massive remodeling of organelles and chromatin, but whether and how these are linked to control neuronal fate commitment remains unknown. We examined and manipulated mitochondria dynamics with high temporal resolution during mouse and human cortical neurogenesis. We reveal that shortly after cortical stem cells have divided, daughter cells that retain high levels of mitochondria fission will become neurons, while those destined to self-renew undergo rapid mitochondria fusion. Induction of mitochondria fusion after mitosis redirects daughter cells towards cell self-renewal, but only during a restricted time window, which is doubled in human cortical stem cells with higher self-renewing potential. Mitochondria dynamics drives neurogenesis through modulation of the NAD+sensor Sirtuin-1, leading to Histone deacetylation and chromatin remodeling necessary for neurogenic conversion. Our data reveal a post-mitotic critical period of neurogenesis, linking mitochondria state with cell fate.
Improved design and analysis of practical minimizers
Authors: Zheng Hongyu , Kingsford Carl , Marçais Guillaume
Published: February 2020

Motivation

Minimizers are methods to samplek-mers from a sequence, with the guarantee that similar set ofk-mers will be chosen on similar sequences. It is parameterized by thek-mer lengthk, a window lengthwand an order on thek-mers. Minimizers are used in a large number of softwares and pipelines to improve computation efficiency and decrease memory usage. Despite the method’s popularity, many theoretical questions regarding its performance remain open. The core metric for measuring performance of a minimizer is the density, which measures the sparsity of sampledk-mers. The theoretical optimal density for a minimizer is 1/w, provably not achievable in general. For givenkandw, little is known about asymptotically optimal minimizers, that is minimizers with densityO(1/w).

Results

We derive a necessary and sufficient condition for existence of asymptotically optimal minimizers. We also provide a randomized algorithm, called the Miniception, to design minimizers with the best theoretical guarantee to date on density in practical scenarios. Constructing and using the Miniception is as easy as constructing and using a random minimizer, which allows the design of efficient minimizers that scale to the values ofkandwused in current bioinformatics software programs.

Availability

Reference implementation of the Miniception and the codes for analysis can be found athttps://github.com/kingsford-group/miniception.

Contact

gmarcais@cs.cmu.edu
Canopy position has a stronger effect than tree species identity on phyllosphere bacterial diversity in a floodplain hardwood forest
Authors: Herrmann Martina , Geesink Patricia , Richter Ronny , Küsel Kirsten
Published: February 2020

The phyllosphere is a challenging microbial habitat in which microorganisms can flourish on organic carbon released by plant leaves but are also exposed to harsh environmental conditions. Here, we assessed the relative importance of canopy position – top, mid, and bottom at a height between 31 m and 20 m – and tree species identity for shaping the phyllosphere microbiome in a floodplain hardwood forest. Leaf material was sampled from three tree species - maple (Acer pseudoplatanus L.), oak (Quercus robur L.), and lime (Tilia cordata MILL.) - at the Leipzig canopy crane facility (Germany). Estimated bacterial species richness (Chao1) and bacterial abundances approximated by quantitative PCR of 16S rRNA genes exhibited clear vertical trends with a strong increase from the top to the mid and bottom position of the canopy. 30 Operational Taxonomic Units (OTUs) formed the core microbiome, which accounted for 77% of all sequence reads. These core OTUs showed contrasting trends in their vertical distribution within the canopy, pointing to different ecological preferences and tolerance to presumably more extreme conditions at the top position of the canopy. Co-occurrence analysis revealed distinct tree species-specific OTU networks, and 55-57% of the OTUs were unique to each tree species. Overall, the phyllosphere microbiome harbored surprisingly high fractions of Actinobacteria of up to 46%. Our results clearly demonstrate strong effects of the position in the canopy on phyllosphere bacterial communities in a floodplain hardwood forest and - in contrast to other temperate or tropical forests - a strong predominance of Actinobacteria.
Forensic implication of cadaveric behaviors of the Japanese pavement ant (Tetramorium tsushimae Emery) attending to rabbit cadavers
Authors: Park Sang‐Hyun , Moon Tae‐Young
Published: February 2020

It is noticed by some researchers that ants may cause some forensic confusion in understanding death scenes. In the aspect of relationship between the ant‐animal decomposition, we observed the behavior of ants congregating on dead animals. Amongst various species of ants visiting dead rabbits, Tetramorium tsushimae Emery was noticeable in producing scratched scars, mounds, and nests around dead rabbits and covering them with soils. The behaviors are likely to interfere or to disturb the usual process of decomposition succession lead by maggots. We discussed that a series of cadaveric behaviors of T. tsushimae may be used as a potential forensic hint not to misunderstand death scenes disturbed by the species.
Food allergy immunotherapy: Oral immunotherapy and epicutaneous immunotherapy
Authors: Kim Edwin H. , Burks Arvil Wesley
Published: February 2020

IgE‐mediated food allergy remains a significant and growing problem across the globe. Of the various treatment modalities, oral immunotherapy (OIT) and epicutaneous immunotherapy (EPIT) have been the best studied. Across various studies of OIT for egg, milk, and peanut allergy, strong levels of desensitization have been shown. With egg and peanut OIT, a limited remission, or sustained unresponsiveness (SU), has further been demonstrated. These advances have been further validated by successful phase 2 and phase 3 studies of peanut OIT. EPIT, using daily administrations of a proprietary patch, demonstrated efficacy as well as safety and tolerability in parallel phase 2 studies; however, its phase 3 study did not meet its primary efficacy outcome. Despite its good track record of desensitization, the safety and tolerability of OIT has remained a question. EPIT, on the other hand, has proven safe and tolerable; however, the adequacy of its desensitization has remained to be determined. As OIT and EPIT continue their march toward regulatory review, optimizations for immunotherapy and novel therapies continue to be developed providing hope for food allergy patients everywhere.
Difficult birth is the main contributor to birth‐related fracture and accidents to other neonatal fractures
Authors: Högberg Ulf , Fellman Vineta , Thiblin Ingemar , Karlsson Ruth , Wester Knut
Published: February 2020

Aim

Specific birth‐related fractures have been studied; underestimates might be a problem. We aimed to assess all fractures diagnosed as birth‐related as well as other neonatal fractures.

Methods

A population‐based study on all infants born in Sweden 1997‐2014; data were retrieved from the Swedish Health Registers (10th version of International Classification of Diseases. Outcome measures were birth‐related fractures (ICD‐10 P‐codes) and other neonatal fractures (ICD‐10 S‐codes).

Results

The overall fracture incidence was 2.9 per 1000 live birth (N = 5336); 92.6% had P‐codes and 7.4% (S‐codes). Some birth‐related fractures were diagnosed beyond the neonatal period. Other neonatal fractures could have been birth‐related. Clavicle fracture (88.8%) was associated with adverse maternal and infant anthropometrics and birth complications. The few neonates with rib fractures all had concomitant clavicle fracture. For skull fractures, a minor part was birth‐related and most were associated with accidents. Half of the long bone fractures were associated with accidents. Birth‐related femur fractures were associated with bone fragility risk factors. Five infants with abuse diagnoses had fractures: skull (4), long bone (2) and rib (1).

Conclusion

Birth‐related and other neonatal fractures are rarely diagnosed. Difficult birth is the main contributor to birth‐related fracture and accidents to other neonatal fractures.
Efficacy gap between phase II and subsequent phase III studies in oncology
Authors: Vreman Rick A. , Belitser Svetlana V. , Mota Ana T.M. , Hövels Anke M. , Goettsch Wim G. , Roes Kit C.B. , Leufkens Hubert G.M. , Mantel‐Teeuwisse Aukje K.
Published: February 2020

Aims

There is a trend for more flexibility in timing of evidence generation in relation to marketing authorization, including the option to complete phase III trials after authorization or not at all. This paper investigated the relation between phase II and III clinical trial efficacy in oncology.

Methods

All oncology drugs approved by the European Medicines Agency (2007–2016) were included. Phase II and phase III trials were matched based on indication and treatment and patient characteristics. Reported objective response rates (ORR), median progression‐free survival (PFS) and median overall survival (OS) were analysed through weighted mixed‐effects regression with previous treatment, treatment regimen, blinding, randomization, marketing authorization type and cancer type as covariates.

Results

A total of 81 phase II‐III matches were identified including 252 trials. Mean (standard deviation) weighted difference (phase III minus II) was −4.2% (17.4) for ORR, 2.1 (6.7) months for PFS and −0.3 (5.1) months for OS, indicating very small average differences between phases. Differences varied substantially between individual indications: from −46.6% to 47.3% for ORR, from −5.3 to 35.9 months for PFS and from −13.3 to 10.8 months for OS. All covariates except blinding were associated with differences in effect sizes for at least 1 outcome.

Conclusions

The lack of marked average differences between phases may encourage decision‐makers to regard the quality of design and total body of evidence instead of differentiating between phases of clinical development. The large variability emphasizes that replication of study findings remains essential to confirm efficacy of oncology drugs and discern variables associated with demonstrated effects.
We are all in this together: The role of individuals’ social identities in problematic engagement with video games and the internet
Authors: Travaglino Giovanni A. , Li Zhuo , Zhang Xingruo , Lu Xian , Choi Hoon‐Seok
Published: February 2020

Individuals’ engagement with video games and the internet features both social and potentially pathological aspects. In this research, we draw on the social identity approach and present a novel framework to understand the linkage between these two aspects. In three samples (Nstudy1 = 304, Nstudy2 = 160, and Nstudy3 = 782) of young Chinese people from two age groups (approximately 20 and 16 years old), we test the associations between relevant social identities and problematic engagement with video games and the internet. Across studies, we demonstrate that individuals’ identification as ‘gamers’ or ‘frequent internet users’ predicts problematic engagement with video games and the internet through stronger perceived social support from such groups. Moreover, we demonstrate that individuals’ identification as ‘students’ (Studies 2–3) is negatively associated with problematic engagement via social support from other students. Finally, in Study 3, we examine the articulation between social support from these three groups and subjective sense of loneliness. Findings indicate that, whereas perceived support from students is negatively associated with loneliness, the association between perceived support from gamers and internet users and loneliness is weaker and positive. Theoretical implications and directions for future research are discussed. Taken together, the studies highlight the importance of considering the social context of individuals’ problematic engagement with technologies, and the role of different group memberships.
Underdogs make an alliance: The co‐experience of rejection promotes cooperation
Authors: Miao Xiao‐Yan , Chan Kai Qin , Gao Cong , Lv Sa‐Sa , Zhu Yuxi , Wang Zuo‐Jun
Published: February 2020

Social rejection research has largely focused on the consequences of rejection when individuals experience rejection alone. Yet little is known about the reaction of those co‐experiencing rejection. We tested the hypothesis that the co‐experience of rejection increases cooperation between the co‐experiencers. Three experiments provided supporting evidence for the hypothesis. The participants cooperated more when they co‐experienced rejection than when they experienced rejection alone. The need to belong mediated the relationship between those co‐experiencing rejection and cooperation. These findings shed light on the factors that initiate the formation of small groups, especially deviant ones.
Characterisation of endogenous Claudin‐1 expression, motility and susceptibility to hepatitis C virus in CRISPR knock‐in cells
Authors: Clément Camille M.H. , Deffieu Maika S. , Dorobantu Cristina M. , Baumert Thomas F. , Ayala‐Nunez Nilda Vanesa , Mély Yves , Ronde Philippe , Gaudin Raphael
Published: February 2020

Background Information

Claudin‐1 (CLDN1) is a four‐span transmembrane protein localised at cell–cell tight junctions (TJs), playing an important role in epithelial impermeability and tissue homoeostasis under physiological conditions. Moreover, CLDN1 expression is up‐regulated in several cancers, and the level of CLDN1 expression has been proposed as a prognostic marker of patient survival.

Results

Here, we generated and characterised a novel reporter cell line expressing endogenous fluorescent levels of CLDN‐1, allowing dynamic monitoring of CLDN‐1 expression levels. Specifically, a hepatocellular carcinoma Huh7.5.1 monoclonal cell line was bioengineered using CRISPR/Cas9 to endogenously express a fluorescent TagRFP‐T protein fused at the N‐terminus of the CLDN1 protein. These cells were proved useful to measure CLDN1 expression and distribution in live cells. However, the cells were resistant to hepatitis C virus (HCV) infection, of which CLDN1 is a viral receptor, while retaining permissiveness to VSV‐G‐decorated pseudoparticles. Nonetheless, the TagRFP‐CLDN1+/+ cell line showed expected CLDN1 protein localisation at TJs and the cell monolayer had similar impermeability and polarisation features as its wild‐type counterpart. Finally, using fluorescence recovery after photobleaching (FRAP) approaches, we measured that the majority of endogenous and overexpressed TagRFP‐CLDN1 diffuses rapidly within the TJ, whereas half of the overexpressed EGFP‐CLDN1 proteins were stalled at TJs.

Conclusions

The Huh7.5.1 TagRFP‐CLDN1+/+ edited cell line showed physiological features comparable to that of non‐edited cells, but became resistant to HCV infection. Our data also highlight the important impact of the fluorescent protein chosen for endogenous tagging.

Significance

Although HCV‐related studies may not be achieved with these cells, our work provides a novel tool to study the cell biology of TJ‐associated proteins and a potential screening strategy measuring CLDN1 expression levels.
Vaccines targeting the primary amino acid sequence and conformational epitope of amyloid‐β had distinct effects on neuropathology and cognitive deficits in EAE/AD mice
Authors: Yu Xiao‐Lin , Zhu Jie , Liu Xiang‐meng , Xu Peng‐xin , Zhang Yue , Liu Rui‐tian
Published: February 2020

Background and Purpose

Immunotherapeutic intervention is one of the most promising strategies for the prevention and treatment of Alzheimer's disease (AD). Although they showed great success in AD mouse models, the clinical trials of many immune approaches failed due to low efficacy and safety. Thus, an animal model which can show the potential side effects of vaccines or antibodies is urgently needed. In this study, we generated EAE/AD mice by crossing APP/PS1 mice with experimental autoimmune encephalomyelitis (EAE) mice. We then investigated the efficacy and safety of two vaccines: the immunogens of which were Aβ1‐42 aggregates (Aβ42 vaccine) and an oligomer‐specific conformational epitope (AOE1 vaccine), respectively.

Experimental Approach

EAE/AD mice were immunized with the Aβ42 vaccine or AOE1 vaccine five times at biweekly intervals. After the final immunization, cognitive function was evaluated by the Morris water maze, Y maze, and object recognition tests. Neuropathological changes in the mouse brains were analysed by immunohistochemistry and ELISA.

Key Results

In contrast to previous findings in conventional AD animal models, Aβ42 immunization promoted neuroinflammation, enhanced Aβ levels and plaque burden, and failed to restore cognitive deficits in EAE/AD mice. By contrast, AOE1 immunization dramatically attenuated neuroinflammation, reduced Aβ levels, and improved cognitive performance in EAE/AD mice.

Conclusion and Implications

These results suggest that the EAE/AD mouse model can exhibit the potential side effects of AD immune approaches that conventional AD animal models fail to display. Furthermore, strategies specifically targeting Aβ oligomers may be safe and show clinical benefit for AD treatment.
The role of astrocytes in oxidative stress of central nervous system: A mixed blessing
Authors: Chen Yaxing , Qin Chen , Huang Jianhan , Tang Xin , Liu Chang , Huang Keru , Xu Jianguo , Guo Gang , Tong Aiping , Zhou Liangxue
Published: February 2020

Central nervous system (CNS) maintains a high level of metabolism, which leads to the generation of large amounts of free radicals, and it is also one of the most vulnerable organs to oxidative stress. Emerging evidences have shown that, as the key homeostatic cells in CNS, astrocytes are deeply involved in multiple aspects of CNS function including oxidative stress regulation. Besides, the redox level in CNS can in turn affect astrocytes in morphology and function. The complex and multiple roles of astrocytes indicate that their correct performance is crucial for the normal functioning of the CNS, and its dysfunction may result in the occurrence and progression of various neurological disorders. To date, the influence of astrocytes in CNS oxidative stress is rarely reviewed. Therefore, in this review we sum up the roles of astrocytes in redox regulation and the corresponding mechanisms under both normal and different pathological conditions.
The utility of elastography during EBUS‐TBNA in a population with a high prevalence of anthracosis
Authors: Abedini Atefeh , Razavi Fatemeh , Farahani Mehrdad , Hashemi Marzieh , Emami Habib , Mohammadi Forouzan , Kiani Arda
Published: February 2020

Background

Ultrasound elastography, is a pioneer sonographic modality that is conducted during endobronchial ultrasound‐guided transbronchial needle aspiration (EBUS‐TBNA) in order to increase the accuracy of sampling location. The current study aims to evaluate the usefulness of elastography during EBUS‐TBNA in a population with a high prevalence of anthracosis.

Methods

This prospective single‐blinded study was performed on 69 lymph nodes (LNs) of patients with mediastinal lymphadenopathy undergoing EBUS‐TBNA and EBUS‐elastography from October 2017 to July 2018. The stiffness level of the tissue was translated into a color to demonstrate the hardness of tissue. Blue and total areas of each section were measured to calculate the hardness of each LN.

Results

Sixty‐nine LNs were evaluated by elastography. Twenty percent of LNs were malignant. There was a statistical difference between malignant and non‐malignant nodes based on color dominancy (P = 0.032). However, with the exclusion of anthracosis nodes from the analysis, the difference was more significant (P < 0.001). Moreover, when the blue dominancy was used as the predictor of malignancy or anthracosis, the results showed a significant correlation (P < 001).

Conclusion

The usefulness of elastography in selecting the hardest area of tissue that is appropriate for diagnosing diseases has been proven previously. Since in countries with a high prevalence of anthracosis, blue color achieved using elastography predicts either malignancy or anthracosis so, cases with blue dominancy of LNs in elastography and the white color in the EBUS‐TBNA indicate anthracosis‐caused calcification should be reconsidered.
An investigation of the association between ADRB2 gene polymorphisms and asthma in Kazakh population
Authors: Akparova Almira , Aripova Akmaral , Abishev Meirzhan , Kazhiyakhmetova Baglan , Pirmanova Ainur , Bersimbaev Rakhmetkazhi
Published: February 2020

Introduction

Beta‐2‐adrenergic receptor (ADRB2) is present in the cells of the respiratory tract, including bronchial smooth muscle cells and bronchial epithelium, and is a target for endogenous catecholamines and drugs used to treat the obstructive lung diseases.

Objectives

This study aimed to investigate the possible association of the Arg16Gly and Gln27Glu polymorphisms of the ADRB2 gene with asthma and its endophenotypes in the Kazakh population.

Methods

A total of 70 asthmatic patients and 80 healthy controls were genotyped for Arg16Gly and Gln27Glu polymorphisms of the ADRB2 gene by using quantitative real‐time polymerase chain reaction. Statistical analysis was performed with the Graph Pad InStat 7 Software.

Results

No associations between the asthma patients and healthy individuals were found when the allele and genotype distribution of Arg16Gly and Gln27Glu single nucleotide polymorphisms were compared. Analysis of the haplotype frequencies showed statistically significant differences between patients with asthma and controls for Arg16Gly/Gln27Gln and Arg16Gly/Gln27Glu haplotypes (odds ratio [OR] = 2.12, 95% confidence interval [CI] = 0.87‐5.16 and OR = 2.25, 95% CI = 0.89‐5.67 respectively). The Arg16 allele and Arg16Arg genotype frequencies were higher in patients with uncontrolled asthma than in controls (χ2 = 5.17, df = 1, P = 0.02 and χ2 = 5.36, df = 1, P = 0.02 respectively).

Conclusion

The results of this study support the possible involvement of Arg16Gly polymorphism in the development of uncontrolled asthma, and indicate that Arg16Gly/Gln27Gln and Arg16Gly/Gln27Glu haplotypes are more common in asthma patients in the Kazakh population.
A learning curve in using orphan liver allografts for transplantation
Authors: Rana Abbas , Joshi Manasi , Price Matthew Brent , Ganni Saif , Bakhtiyar Syed S. , Vierling John M. , Galvan Nhu Thao , Cotton Ronald T. , O'Mahony Christine A. , Kanwal Fasiha , Goss John A.
Published: February 2020

Given the critical shortage of donor livers, marginal liver allografts have potential to increase donor supply. We investigate trends and long‐term outcomes of liver transplant using national share allografts transplanted after rejection at the local and regional levels. We studied a cohort of 75 050 candidates listed in the Organ Procurement and Transplantation Network for liver transplantation between 2002 and 2016. We compared patients receiving national share and regional/local share allografts from 2002‐2006, 2007‐2011, and 2012‐2016, performing multivariate Cox regression for graft survival. Recipient and center‐level covariates that were not significant (P < .05) were removed. Graft survival of national share allografts improved over time. National share allografts had a 26% increased risk for graft failure in 2002‐2006 but no impact on graft survival in 2007‐2011 and 2012‐2016. The cold ischemia time (CIT) of national share allografts decreased from 10.4 to 8.0 hours. We demonstrate that CIT had significant impact on graft survival using national share allografts (CIT <6 hours: hazard ratio 0.75 and CIT >12 hours: hazard ratio 1.25). Despite a trend toward sicker recipients and poorer quality allografts, graft survival outcomes using national share allografts have improved to benchmark levels. Reduction in cold ischemia time is a possible explanation.
Nevus psiloliparus: Newly described histopathological features from transverse sections
Authors: Roman Jorge , Taylor Nicholas A. , Oza Vikash S. , Kim Randie H.
Published: February 2020

Nevus psiloliparus is a rare fatty tissue nevus that is a marker for encephalocraniocutaneous lipomatosis, a neurocutaneous syndrome with ocular and central nervous system anomalies. Clinically, nevus psiloliparus is often described as a congenital alopecia and appears as an irregularly shaped, circumscribed area of alopecia on the scalp. Histopathology demonstrates a near‐complete absence of mature hair follicles with preservation of arrector pili muscles and mature adipocytes within the dermis. The pathogenesis of nevus psiloliparus may be related to mosaic mutations in fibroblast growth factor receptor 1. Herein we report the histopathological features of a nevus psiloliparus in an 11‐year‐old girl diagnosed from transverse sections, which show “shadow” follicular units characterized by columns of loosely arranged collagen and a relative paucity of elastic fibers.
HERGEST CROFT GARDEN, AN INTERNATIONAL PLANT COLLECTION IN HEREFORDSHIRE
Authors: Lawrence  , Banks Elizabeth
Published: February 2020

The history of the garden at Hergest Croft is given, and the varied sources of some of the most interesting trees are described.
THE MANAGEMENT OF THE WOODY PLANT COLLECTION AT HERGEST CROFT
Authors: Griffiths Rowan
Published: February 2020

The management of the collection at Hergest Croft is described. The effects and control of pests such as deer and grey squirrels are discussed. Endangered species in the collection are assessed.
921. CALYCANTHUS CHINENSIS
Authors: Lancaster Roy , Rix Martyn
Published: February 2020

Calycanthus chinensis (W.C. Cheng & S.Y. Chang) W.C. Cheng & S.Y. Chang ex P.T. Li, is illustrated. Its distribution in China and instructions for its cultivation are given. The other members of the genus Calycanthus are discussed.
922. LINDERA OBTUSILOBA
Authors: Lancaster Roy , Rix Martyn
Published: February 2020

Lindera obtusiloba Blume is illustrated. Its distribution in China, Japan and Korea is reported. The synonyms are listed and discussed. Instructions for its cultivation are given.
925. GRIFFITHARIA HEDLUNDII
Authors: Rushforth Keith
Published: February 2020

Griffitharia hedlundii (C.K. Schneider) Rushforth (syn. Sorbus hedlundii C.K. Schneider) is illustrated. Its distribution and ecology is reported. The synonyms are listed and described. Instructions for its cultivation are given.
926. THOMSONARIA ELEONORAE
Authors: Rushforth Keith
Published: February 2020

Thomsonaria eleonorae is illustrated. Its distribution and ecology is reported. The synonyms are listed and described. Instructions for its cultivation are given.
929. ACER MORIFOLIUM
Authors: Crowley Dan
Published: February 2020

Acer morifolium Koidz. is illustrated. Its history, ecology and distribution in Japan, are noted. Instructions for its cultivation are given.
931. RHODODENDRON SUOILENHENSE
Authors: Rushforth Keith , Huong Nguyen Thi Thanh , Chamberlain David
Published: February 2020

A new species of Rhododendron, R. suoilenhense D.F. Chamberlain, Nguyen Thi Thanh Huong & Rushforth, is described from northern Vietnam and illustrated. Its discovery and distribution are noted. Suggestions for its cultivation are given.
Non‐malignant fibroblastic/myofibroblastic tumors in pediatric age group: Clues and pitfalls to the cytological diagnosis
Authors: Agarwal Harshita , Singh Lavleen , Gupta Natasha , Sengar Mamta
Published: February 2020

Introduction

Fibroblastic/myofibroblastic tumors constitute 12% of all pediatric soft tissue tumors with the majority of them belonging to the benign and intermediate prognostic categories. They are often misdiagnosed owing to their variable clinical presentation and unusual microscopic features. The diagnosis, specially cytological diagnosis of benign and intermediate categories is difficult due to paucity of cellular component and increased amount of extracellular matrix as compared to malignant ones. We hereby discuss the Fine needle aspiration cytology (FNAC) findings of non‐malignant fibroblastic/myofibroblastic lesion in the pediatric age group encountered at our institute.

Methods

All the benign and intermediate fibroblastic/myofibroblastic/fibroadipocytic lesions (age 0‐12 years) diagnosed on FNAC over a period of 3½ years (Jan 2016‐ July 2019), with availability of corresponding histopathology were included in the study.

Results

A total of seven pediatric benign and intermediate fibroblastic/myofibroblastic lesions with histopathological confirmation were identified which included Infantile digital fibromatosis (IDF) (n = 2), Lipofibromatosis (n = 1), Fibrous hamartoma of infancy (FHI) (n = 1), Fibromatosis colli (FC) (n = 2) and myofibroma/myofibromatosis (n = 1). FNAC smears were mainly paucicellular with presence of benign spindle shaped cells in a collagenous stroma common to almost all the cases. A few additional findings such as degenerated skeletal muscle fibres, muscle giant cells and mature adipose tissue were also present in some cases.

Conclusion

Fibroblastic/myofibroblastic tumors although uncommon, form an important category that must be considered in the differential diagnosis of pediatric soft tissue tumors. FNAC cytology features when assessed in a proper clinical setting (specially the age and site of presentation) are helpful in suggesting probable preoperative diagnosis in these lesions.
What are the characteristics of the best type 1 diabetes patient education programmes (from diagnosis to long‐term care), do they improve outcomes and what is required to make them more effective?
Authors: Heller S. R. , Gianfrancesco C. , Taylor C. , Elliott J.
Published: February 2020

The last 20 years have witnessed a marked change in approaches to the management of type 1 diabetes in the UK. This is exemplified by National Institute of Health and Care Excellence (NICE) guidance which acknowledges that reaching and maintaining target glucose depends on people with type 1 diabetes effectively implementing flexible intensive insulin therapy. The guidance emphasizes that successful self‐management requires the acquisition of complex skills and is best achieved by participation in high‐quality structured education. Controlled trials and other research have shown that programmes teaching self‐management can lower glucose levels while reducing hypoglycaemia, improve psychological outcomes and are highly cost‐effective. An important principle of successful programmes is therapeutic education in which learning becomes a partnership between the professional and the person with diabetes who learns to fit diabetes into his/her everyday life. Other recommended elements of programmes include a written curriculum, group teaching by a professional multidisciplinary team and quality assurance. Yet many participants struggle post‐course to implement and maintain skills, and overall HbA1c levels, particularly in the UK, remain far from target. Recent studies have identified the barriers to sustained effective self‐management and concluded that even high‐quality programmes generally lack critical components. These include incorporating evidence from behaviour change research, exploiting the promise of new technologies in reducing the burden of self‐management, and providing structured professional support once people have completed the training. Studies are currently underway to evaluate structured training courses which have added these elements and examine whether they can lower glucose to levels closer to target without impairing quality of life.