bluuebunny/crossref_metadata_2025 huggingface dataset:
Vitamin D Enhances Radiosensitivity of Colorectal Cancer by Reversing Epithelial-Mesenchymal Transition
Authors: Yu Xinyue , Wang Qian , Liu Baocai , Zhang Ning , Cheng Guanghui
Published: August 2021

Colorectal cancer (CRC) is often resistant to conventional therapies. Previous studies have reported the anticancer effects of vitamin D in several cancers, its role in radiotherapy (RT) remains unknown. We found that 1α, 25-dihydroxyvitamin D3 (VD3), the biologically active form of vitamin D, had antitumor effect on CRC and sensitized CRC cells to ionizing radiation (IR). VD3 demonstrated synergistic effect in combination with IR, which were detected by colony formation and cell proliferation assay. Radiosensitivity restoration induced by VD3 was associated with a series of phenotypes, including apoptosis, autophagy, and epithelial-mesenchymal transition (EMT). Using proteomics, “regulation of cell migration” and “cadherin” were found to be obviously enriched GO terms. Moreover, cystatin D and plasminogen activator inhibitor-1 (PAI-1), the differentially expressed proteins, were associated with EMT. Next, we confirmed the contributions of these two genes in enhancing IR sensitivity of CRC cells upon inhibition of EMT. As determined by proteomics, the mechanism underlying such sensitivity involved partially block of JAK/STAT3 signaling pathway. Furthermore, VD3 also elicited sensitization to RT in xenograft CRC models without additional toxicity. Our study revealed that VD3 was able to act in synergy with IR both in vitro and in vivo and could also confer radiosensitivity by regulating EMT, thereby providing a novel insight for elevating the efficacy of therapeutic regimens.
CTRP1 Attenuates Cerebral Ischemia/Reperfusion Injury via the PERK Signaling Pathway
Authors: Fei Huizhi , Xiang Pu , Luo Wen , Tan Xiaodan , Gu Chao , Liu Maozhu , Chen Mengyuan , Wang Qiong , Yang Junqing
Published: August 2021

Cerebral ischemic stroke is one of the leading causes of death worldwide. Previous studies have shown that circulating levels of CTRP1 are upregulated in patients with acute ischemic stroke. However, the function of CTRP1 in neurons remains unclear. The purpose of this study was to explore the role of CTRP1 in cerebral ischemia reperfusion injury (CIRI) and to elucidate the underlying mechanism. Middle cerebral artery occlusion/reperfusion (MCAO/R) and oxygen–glucose deprivation/reoxygenation (OGD/R) models were used to simulate cerebral ischemic stroke in vivo and in vitro, respectively. CTRP1 overexpression lentivirus and CTRP1 siRNA were used to observe the effect of CTRP1 expression, and the PERK selective activator CCT020312 was used to activate the PERK signaling pathway. We found the decreased expression of CTRP1 in the cortex of MCAO/R-treated rats and OGD/R-treated primary cortical neurons. CTRP1 overexpression attenuated CIRI, accompanied by the reduction of apoptosis and suppression of the PERK signaling pathway. Interference with CTRP1 expression in vitro aggravated apoptotic activity and increased the expression of proteins involved in the PERK signaling pathway. Moreover, activating the PERK signaling pathway abolished the protective effects of CTRP1 on neuron injury induced by CIRI in vivo and in vitro. In conclusion, CTRP1 protects against CIRI by reducing apoptosis and endoplasmic reticulum stress (ERS) through inhibiting the PERK-dependent signaling pathway, suggesting that CTRP1 plays a crucial role in the pathogenesis of CIRI.
SNAI2-Induced CircMTO1 Promotes Cell Proliferation and Inhibits Apoptosis Through the miR-320b/MCL1 Axis in Human Granulosa-Like Tumor Cells
Authors: Duan Jie , Cai Hongning , Huang Yanming , Shi Liangyan
Published: August 2021

Polycystic ovary syndrome (PCOS), one of the most common types of endocrine diseases, is characterized by a high prevalence among women of reproductive-age. However, its pathogenesis and molecular mechanisms remain unclear. CircMTO1 has been reported to participate in numerous biological processes, but, its role in PCOS progression remains unknown. In the current study, we elucidated the expression and circRNA characterization of circMTO1 in human granulosa-like tumor cells. We found that circMTO1 knockdown promoted human granulosa-like tumor cell proliferation and inhibited its apoptosis rate. Next, we explored the underlying molecular mechanisms by using a series of experiments. Our results revealed the effect of the novel circMTO1/miR-320b/MCL1 axis in human granulosa-like tumor cells. Furthermore, we found that the expression of circMTO1 was induced by Snail family transcriptional repressor 2 (SNAI2) in human granulosa-like tumor cells. Our results may provide potential targets for PCOS research and a novel direction for the diagnosis and treatment of PCOS.
Redefining the Distributional Boundaries and Phylogenetic Relationships for Ctenomids From Central Argentina
Authors: Carnovale Cecilia Soledad , Fernández Gabriela Paula , Merino Mariano Lisandro , Mora Matías Sebastián
Published: August 2021

With about 68 recognized living species, subterranean rodents of the genus Ctenomys are found in a multiplicity of habitats, from the dunes of the Atlantic coast to the Andes Mountains, including environments ranging from humid steppes of Pampas to the dry deserts of Chaco region. However, this genus needs an exhaustive reevaluation of its systematic and phylogenetic relationships regarding the different groups that compose it. This knowledge is essential to propose biodiversity conservation strategies both at species level and at higher hierarchical levels. In order to clarify the taxonomy and the recent evolutionary history from populations of Ctenomys in the Pampas region, Argentina, phylogenetic relationships among them were evaluated using mitochondrial DNA sequences: gene encoding cytochrome b protein (1,140 bp) and the non-coding D-loop region (434 bp). To infer the divergence times inside the Ctenomys clade, a Bayesian calibrate tree using fossil remains data from different families within Caviomorpha was performed at first. Secondly, that calibration data was used as priors in a new Bayesian phylogenetic inference within the genus Ctenomys. This phylogenetic tree emphasized on species currently distributed on the Pampas region, more precisely considering both the talarum and mendocinus groups. Bayesian inferences (BI) were integrated with the results of a Maximum Likelihood approach (ML). Based on these results, the distributional limits of the mendocinus and talarum groups appear to be related to the physiognomy of the Pampas region soils. On the other hand, the validity of C. pundti complex as a differentiated species of C. talarum is debated. According to previous evidence from morphological and chromosomal studies, these results show a very low divergence between those species that originally were classified within the talarum group. Mitochondrial DNA sequences from populations associated with these putative species have not recovered as reciprocal monophyletic groups in the phylogenetic analyses. In conclusion, C. talarum and C. pundti complex might be considered as the same biological species, or lineages going through a recent or incipient differentiation process. The results obtained in this study have important implications for conservation policies and practices, since both species are currently categorized as Vulnerable and Endangered, respectively.
Antibody Response to SARS-CoV-2 Membrane Protein in Patients of the Acute and Convalescent Phase of COVID-19
Authors: Jörrißen Philipp , Schütz Paula , Weiand Matthias , Vollenberg Richard , Schrempf Inga Marie , Ochs Kevin , Frömmel Christopher , Tepasse Phil-Robin , Schmidt Hartmut , Zibert Andree
Published: August 2021

Understanding the course of the antibody response directed to individual epitopes of SARS-CoV-2 proteins is crucial for serological assays and establishment of vaccines. Twenty-one synthetic peptides were synthesized that have ten amino acids overlap and cover the complete membrane (M) protein. Plasma samples from 32 patients having acute disease and 30 patients from the convalescent phase were studied. Only peptide M01 (aa 1–20) and to a lesser extent peptide M21 (aa 201–222) showed specific reactivity as compared to historical control plasma samples. Peptide M01 was recognized by IgM- (71.9%) and IgG-specific antibodies (43.8%) during the acute phase as early as day 8 PIO. In a longitudinal analysis, a higher reactivity was observed for the IgM response directed to peptide M01 following day 20 PIO as compared to earlier time points of the acute phase. In the convalescent phase, antibody reactivity to the two M-specific peptides was significantly lower (<30% seropositivity). A fusion protein encoding major parts of RBD also showed higher rates of recognition during acute (50.0%) and lower rates in the convalescent phase (23.3%). Taken together, our results suggest that during the acute phase of COVID-19 antibodies are raised to two linear epitopes of the SARS-CoV-2 M protein, located at the very N- and C-termini, showing almost similar levels of reactivity as immunodominant linear epitopes derived from the spike and nucleocapsid protein. Anti-M is also present in the convalescent phase of COVID-19 patients, however at lower levels, with the N-terminus of the M protein as a preferred target.
Distribution of Cetaceans in the Canary Islands (Northeast Atlantic Ocean): Implications for the Natura 2000 Network and Future Conservation Measures
Authors: Herrera Inma , Carrillo Manuel , Cosme de Esteban Marcial , Haroun Ricardo
Published: August 2021

The waters of the Canary Islands are considered a hotspot for marine biodiversity, especially regarding cetacean species. Based on this fact, this study pays attention to the spatial distribution pattern of cetacean species and the conservation role of the Natura 2000 Network, a set of Special Areas of Conservation (SACs), which were defined mainly based on data compiled in 1996, under the framework of the European Habitats Directive. In recent years, the declaration of conservation areas for cetaceans between the Tenerife—La Gomera Islands by two global conservation programs, Mission Blue (“Hope Spots”) and Whale Heritage Site (“Whale Sanctuary”) sent clear signals of scientific and social interest to promote better protection of the cetacean species in the Canary Islands. The main aim of the designated SACs is the conservation of its biological and ecological diversity, ensuring the long-term survival of the target species in the waters around islands. In this case, the enactment of the SACs was based only on the sparse data available for the common bottlenose dolphin, Tursiops truncatus. This study shows that the spatial distribution of cetaceans in the Canary archipelago generated from a large database of cetacean sightings, from 2007 to 2018. The results obtained show the main marine areas where the different cetacean species are distributed around the different islands of the archipelago. The spatial distribution maps of the cetacean species, when compared with the existing SACs of the Natura 2000, show the need to extend these SACs into the open sea to include more cetacean species and a larger number of individuals for better conservation of the endangered marine mammals. As a consequence, some suggestions were proposed to improve and update the role of SACs in European Northeast Atlantic waters as a key environmental tool for cetacean conservation. The data supporting the recent declarations of these two new milestones the “Hope Spot” and the “Whale Sanctuary” enhance more keystone information to promote a large marine protected area in the Eastern Atlantic Ocean, such as the “Macaronesian Biodiversity and Ecological Migration Corridor for Cetaceans,” a conservation figure that has been already proposed in the scientific literature as a deserving candidate of governmental regulations and policies by Portugal and Spain; it would also require joint cross-border cooperation efforts for marine spatial planning.
Application of 3D Balanced Growth Theory to the Formation of Bulk Amorphous Alloys
Authors: Wu YaQi , Zhang Yong , Zhang Tao
Published: August 2021

Since the emergence of amorphous alloys as a new class of materials, efficiency improvements have been made in optimizing the fabrication process, the mechanization of alloy formation, and the size of the alloys themselves. Amorphous alloys have been used in precision instruments as they possess excellent magnetic properties, corrosion resistance, wear resistance, high strength, hardness, toughness, high electrical resistivity, and electromechanical coupling properties. Because their hysteresis losses are lower than those of traditional transformer cores, the conversion efficiency of equipment has been significantly improved, thereby saving energy and protecting the environment. Hence, amorphous iron cores have replaced traditional materials. Amorphous alloys also show excellent performance as anti-corrosion and wear-resistant coatings. The process of preparing amorphous alloys starts with an amorphous alloy film obtained by evaporation deposition and then proceeds to the use of a high cooling rate ribbon spinning method to finally obtain a thin strip of an amorphous alloy. A widely used method of copper mold suction casting is then used to prepare the bulk amorphous alloy. The sizes of amorphous alloys have been continually increasing, which has resulted in increasingly serious challenges, such as cooling rate and thermal stability limitations. In addition, crystals can form at low cooling rates. The latent heat of crystallization is released when crystals are formed, which causes damage to the amorphous area so that the size of amorphous alloys is reduced. Because of these difficulties, new processes that eliminate the cooling rate gradient, such as 3D additive manufacturing, ultrasonic production, and mold design, combined with the concept of “entropy control” component design and the economic theory of “balanced development,” lead to a three-dimensional bulk amorphous alloy being proposed. The theory of balanced growth provides a new concept for the development and application of bulk amorphous alloys. This review offers a retrospective view of recent studies of amorphous alloys and provides a description of the formation of amorphous alloys and amorphous phases and the criteria required to predict the successful formation of amorphous alloys. Then, we address the problem of size limitation confronting current production methods. The three-dimensional balanced growth theory of bulk amorphous alloys was formulated from a flexible adaptation of the balanced growth theory of economics. We have confidence that the production and development of bulk amorphous alloys have a bright future.
Case Report: A Novel Variant c.2262+3A>T of the SCN5A Gene Results in Intron Retention Associated With Incessant Ventricular Tachycardias
Authors: Yin Jie , Zhou Jia , Chen Jinlong , Xu Ting , Zhang Zhongman , Zhang Han , Yuan Chang , Cheng Xueying , Qin Yuming , Zheng Bixia , Wang Chunli , Yang Shiwei , Jia Zhanjun
Published: August 2021

Objective: Voltage-gated sodium channel Nav1.5 encoded by the SCN5A gene plays crucial roles in cardiac electrophysiology. Previous genetic studies have shown that mutations in SCN5A are associated with multiple inherited cardiac arrhythmias. Here, we investigated the molecular defect in a Chinese boy with clinical manifestations of arrhythmias.

Methods: Gene variations were screened using whole-exome sequencing and validated by direct Sanger sequencing. A minigene assay and reverse transcription PCR (RT-PCR) were performed to confirm the effects of splice variants in vitro. Western blot analysis was carried out to determine whether the c.2262+3A>T variant produced a truncated protein.

Results: By genetic analysis, we identified a novel splice variant c.2262+3A>T in SCN5A gene in a Chinese boy with incessant ventricular tachycardias (VT). This variant was predicted to activate a new cryptic splice donor site and was identified by in silico analysis. The variant retained 79 bp at the 5′ end of intron 14 in the mature mRNA. Furthermore, the mutant transcript that created a premature stop codon at 818 amino acids [p.(R818*)] could be produced as a truncated protein.

Conclusion: We verified the pathogenic effect of splicing variant c.2262+3A>T, which disturbed the normal mRNA splicing and caused a truncated protein, suggesting that splice variants play an important role in the molecular basis of early onset incessant ventricular tachycardias, and careful molecular profiling of these patients will be essential for future effective personalized treatment options.
Patient Reported Experiences and Delays During the Diagnostic Pathway for Pulmonary Fibrosis: A Multinational European Survey
Authors: van der Sar Iris G. , Jones Steve , Clarke Deborah L. , Bonella Francesco , Fourrier Jean Michel , Lewandowska Katarzyna , Bermudo Guadalupe , Simidchiev Alexander , Strambu Irina R. , Wijsenbeek Marlies S. , Parfrey Helen
Published: August 2021

Introduction: Pulmonary fibrosis includes a spectrum of diseases and is incurable. There is a variation in disease course, but it is often progressive leading to increased breathlessness, impaired quality of life, and decreased life expectancy. Detection of pulmonary fibrosis is challenging, which contributes to considerable delays in diagnosis and treatment. More knowledge about the diagnostic journey from patients' perspective is needed to improve the diagnostic pathway. The aims of this study were to evaluate the time to diagnosis of pulmonary fibrosis, identify potential reasons for delays, and document patients emotions.

Methods: Members of European patient organisations, with a self-reported diagnosis of pulmonary fibrosis, were invited to participate in an online survey. The survey assessed the diagnostic pathway retrospectively, focusing on four stages: (1) time from initial symptoms to first appointment in primary care; (2) time to hospital referral; (3) time to first hospital appointment; (4) time to final diagnosis. It comprised open-ended and closed questions focusing on time to diagnosis, factors contributing to delays, diagnostic tests, patient emotions, and information provision.

Results: Two hundred and seventy three participants (214 idiopathic pulmonary fibrosis, 28 sarcoidosis, 31 other) from 13 countries responded. Forty percent of individuals took ≥1 year to receive a final diagnosis. Greatest delays were reported in stage 1, with only 50.2% making an appointment within 3 months. For stage 2, 73.3% reported a hospital referral within three primary care visits. However, 9.9% reported six or more visits. After referral, 76.9% of patients were assessed by a specialist within 3 months (stage 3) and 62.6% received a final diagnosis within 3 months of their first hospital visit (stage 4). Emotions during the journey were overall negative. A major need for more information and support during and after the diagnostic process was identified.

Conclusion: The time to diagnose pulmonary fibrosis varies widely across Europe. Delays occur at each stage of the diagnostic pathway. Raising awareness about pulmonary fibrosis amongst the general population and healthcare workers is essential to shorten the time to diagnosis. Furthermore, there remains a need to provide patients with sufficient information and support at all stages of their diagnostic journey.
Occurrence, Distribution, Evolutionary Relationships, Epidemiology, and Management of Orthotospoviruses in China
Authors: Zhang Zhongkai , Zheng Kuanyu , Zhao Lihua , Su Xiaoxia , Zheng Xue , Wang Tiantian
Published: August 2021

Orthotospoviruses are responsible for serious crop losses worldwide. Orthotospoviral diseases have spread rapidly in China over the past 10 years and are now found in 19 provinces. Currently, 17 Orthotospovirus species have been reported in China, including eight newly identified species from this genus. The number of new highly pathogenic Orthotospovirus strains or species has increased, likely because of the virus species diversity, the wide range of available hosts, adaptation of the viruses to different climates, and multiple transmission routes. This review describes the distribution of Orthotospovirus species, host plants, typical symptoms of infection under natural conditions, the systemic infection of host plants, spatial clustering characteristics of virus particles in host cells, and the orthotospoviral infection cycle in the field. The evolutionary relationships of orthotospoviruses isolated from China and epidemiology are also discussed. In order to effectively manage orthotospoviral disease, future research needs to focus on deciphering the underlying mechanisms of systemic infection, studying complex/mixed infections involving the same or different Orthotospovirus species or other viruses, elucidating orthotospovirus adaptative mechanisms to multiple climate types, breeding virus-resistant plants, identifying new strains and species, developing early monitoring and early warning systems for plant infection, and studying infection transmission routes.
Time-Specific Pattern of Iron Deposition in Different Regions in Parkinson's Disease Measured by Quantitative Susceptibility Mapping
Authors: Fu Xiaodi , Deng Wenbin , Cui Xiangqin , Zhou Xiao , Song Weizheng , Pan Mengqiu , Chi Xiao , Xu Jinghui , Jiang Ying , Wang Qun , Xu Yunqi
Published: August 2021

Studies have shown the spatial specificity of cranial iron deposition in different regions in Parkinson's disease (PD). However, the time-specific patterns of iron deposition are not yet clear. The purpose of this study was to investigate the time pattern of iron variations and its clinical relevance in multiple gray matter nuclei in PD using quantitative susceptibility mapping (QSM). Thirty controls and 33 PD patients were enrolled, namely, 11 cases of early stage of PD (ESP) and 22 cases of advanced stage of PD (ASP) according to the Hoehn-Yahr stages. The iron content in the subcortical nuclei covering substantia nigra (SN), red nucleus (RN), head of the caudate nucleus (CN), globus pallidus (GP), and putamen (PT) was measured using QSM, and the clinical symptoms of PD were evaluated by various rating scales. The QSM values in SN, RN, GP, and PT significantly increased in PD patients compared with the controls. Further subgroup comparison with the controls indicated that the iron content in SN and GP (paleostriatum) gradually elevated in the whole disease duration and was related to clinical features. While the iron content in RN and PT (neostriatum) only elevated significantly in ESP patients, further iron deposition was not obvious in ASP patients. Our study confirmed that QSM could be used as a disease biomarker and could be suitable for longitudinal monitoring. However, considering the temporal characteristics of iron deposition in neostriatum, iron deposition in the neostriatum should be paid more attention in the early stage of the disease, even in the preclinical stage, in future research.
A Bulk Retrospective Study of Robot-Assisted Stereotactic Biopsies of Intracranial Lesions Guided by Videometric Tracker
Authors: Liu Huan-Guang , Liu Yu-Ye , Zhang Hua , Meng Fan-Gang , Zhang Kai , Zhu Guan-Yu , Chen Ying-Chuan , Liu De-Feng , Zhang Jian-Guo , Yang An-Chao
Published: August 2021

Background: Biopsies play an important role in the diagnosis of intracranial lesions, and robot-assisted procedures are increasingly common in neurosurgery centers. This research investigates the diagnoses, complications, and technology yield of 700 robotic frameless intracranial stereotactic biopsies conducted with the Remebot system.

Method: This research considered 700 robotic biopsies performed between 2016 and 2020 by surgeons from the Department of Functional Neurosurgery in Beijing's Tiantan Hospital. The data collected included histological diagnoses, postoperative complications, operation times, and the accuracy of robotic manipulation.

Results: Among the 700 surgeries, the positive rate of the biopsies was 98.2%. The most common histological diagnoses were gliomas, which accounted for 62.7% of cases (439/700), followed by lymphoma and germinoma, which accounted for 18.7% (131/700) and 7.6% (53/700). Bleeding was found in 14 patients (2%) by post-operation computed tomography scans. A total of 29 (4.14%) patients had clinical impairments after the operation, and 9 (1.29%) experienced epilepsy during the operation. The post-biopsy mortality rate was 0.43%. Operation time—from marking the cranial point to suturing the skin—was 16.78 ± 3.31 min (range 12–26 min). The target error was 1.13 ± 0.30 mm, and the entry point error was 0.99 ± 0.24 mm.

Conclusion: A robot-assisted frameless intracranial stereotactic biopsy guided by a videometric tracker is an efficient, safe, and accurate method for biopsies.
ANK3 Gene Polymorphism Rs10994336 Influences Executive Functions by Modulating Methylation in Patients With Bipolar Disorder
Authors: Tang Lili , Liu Juan , Zhu Yue , Duan Jia , Chen Yifan , Wei Yange , Gong Xiaohong , Wang Fei , Tang Yanqing
Published: August 2021

Background: A large body of evidence suggests that epigenetic modification including DNA methylation plays a critical role in BD's pathogenesis while the identification of methylation quantitative trait loci (meQTLs) shed light on the interpretation of the function of genetic variants in non-coding regions. The intronic single nucleotide polymorphism (SNP) rs10994336 within the ANK3 has emerged as one of the most replicated risk variants for bipolar disorder (BD) in genome-wide association studies. Whether rs10994336 functions as a meQTL to mediate the association between genotype and phenotype remains unclear.

Method: A total of 154 patients with BD and 181 healthy controls (HC) were recruited. The genotypes of rs10994336 and methylation levels of CpG sites within ANK3 were tested. Executive functions were assessed using a computerized version of the Wisconsin Card Sorting Test (WCST).

Results: Bipolar disorder patients with the risk-T allele of rs10994336 scored lower on tests of executive function compared to homozygous CC carriers, after controlling for age, gender, and education level. No significant difference was found in HC individuals. The risk-T allele is associated with a lower methylation level of CpG site cg02172182 in HC after multiple corrections and replicated in the BD group in the same direction. Further mediation analysis revealed that the cg02172182 methylation significantly mediated the association between the polymorphism rs10994336 and PE index of WCST in patients with BD.

Conclusion: Our study suggests that BD-related genetic variant rs10994336 in ANK3 impacts executive functions by modulating ANK3 methylation, supporting the theory that methylation acts as a mediator between genotype and phenotype.
Pharmacological Manipulation of Wnt/β-Catenin Signaling Pathway in Human Neural Precursor Cells Alters Their Differentiation Potential and Neuronal Yield
Authors: Telias Michael , Ben-Yosef Dalit
Published: August 2021

The canonical Wnt/β-catenin pathway is a master-regulator of cell fate during embryonic and adult neurogenesis and is therefore a major pharmacological target in basic and clinical research. Chemical manipulation of Wnt signaling duringin vitroneuronal differentiation of stem cells can alter both the quantity and the quality of the derived neurons. Accordingly, the use of Wnt activators and blockers has become an integral part of differentiation protocols applied to stem cells in recent years. Here, we investigated the effects of the glycogen synthase kinase-3β inhibitor CHIR99021, which upregulates β-catenin agonizing Wnt; and the tankyrase-1/2 inhibitor XAV939, which downregulates β-catenin antagonizing Wnt. Both drugs and their potential neurogenic and anti-neurogenic effects were studied using stable lines human neural precursor cells (hNPCs), derived from embryonic stem cells, which can be induced to generate mature neurons by chemically-defined conditions. We found that Wnt-agonism by CHIR99021 promotes induction of neural differentiation, while also reducing cell proliferation and survival. This effect was not synergistic with those of pro-neural growth factors during long-term neuronal differentiation. Conversely, antagonism of Wnt by XAV939 consistently prevented neuronal progression of hNPCs. We show here how these two drugs can be used to manipulate cell fate and how self-renewing hNPCs can be used as reliable humanin vitrodrug-screening platforms.
The Heparan Sulfate Binding Peptide in Tumor Progression of Triple-Negative Breast Cancer
Authors: Melo Carina Mucciolo , Wang Huawei , Fujimura Ken , Strnadel Jan , Meneghetti Maria Cecília Zorél , Nader Helena Bonciani , Klemke Richard L. , Pinhal Maria Aparecida Silva
Published: August 2021

Angiogenesis is the formation of new vessels from pre-existing vasculature. The heparan sulfate chains from endothelial cell proteoglycans interact with the major angiogenic factors, regulating blood vessels´ formation. Since the FDA´s first approval, anti-angiogenic therapy has shown tumor progression inhibition and increased patient survival. Previous work in our group has selected an HS-binding peptide using a phage display system. Therefore, we investigated the effect of the selected peptide in angiogenesis and tumor progression. The HS-binding peptide showed a higher affinity for heparin N-sulfated. The HS-binding peptide was able to inhibit the proliferation of human endothelial umbilical cord cells (HUVEC) by modulation of FGF-2. It was verified a significant decrease in the tube formation of human endothelial cells and capillary formation of mice aorta treated with HS-binding peptide. HS-binding peptide also inhibited the formation of sub-intestinal blood vessels in zebrafish embryos. Additionally, in zebrafish embryos, the tumor size decreased after treatment with HS-binding peptide.
Wrist, but Not Back, Isometric Contraction Induced Widespread Hypoalgesia in Healthy Participants
Authors: Mailloux Catherine , Wideman Timothy H. , Massé-Alarie Hugo
Published: August 2021

Objective: Exercise may reduce pain sensitivity. This phenomenon called exercise-induced hypoalgesia is observed in different types of exercises and involves the activation of endogenous pain modulation systems. Although the effect of limb exercise on pain sensitivity has often been tested, few studies explored the impact of back exercises that are often used to treat low back pain. The main objective is to measure the effect of back-muscle exercise on pain sensitivity and compare it to the effect of a limb-muscle exercise.

Methods: Twenty-three participants who were pain-free performed a 4-min wrist flexion isometric contraction followed by a 4-min low back extension, separated by a 20-min break. Pressure pain thresholds were tested at two low back (S1 spinous process, lumbar erector spinae muscle) and two wrist (capitate bone, wrist flexor muscles) sites before and after each exercise. For each exercise, sites were considered as remote or local in relation to the muscles contracted during the exercise. An independent sample of 11 participants was recruited to confirm the influence of low back extension on pain sensitivity.

Results: Wrist exercise induced a larger increase in pain sensitivity than back exercise at the remote site. Only wrist exercise induced a hypoalgesia effect at both the local and the remote sites. Back exercise induced a similar effect in the independent sample.

Conclusions: This study showed that back and wrist exercises induced a distinct effect on pain sensitivity in participants who were pain-free. The wrist exercise induced a systemic reduction in pain sensitivity (locally and remotely), whereas the back exercise did not. This differential effect may be present because wrist exercise induced most fatigue compared with the back exercise.
Epidemiology of Bleeding in Critically Ill Children
Authors: Sequeira Jake , Nellis Marianne E. , Karam Oliver
Published: August 2021

Objective: Bleeding can be a severe complication of critical illness, but its true epidemiologic impact on children has seldom been studied. Our objective is to describe the epidemiology of bleeding in critically ill children, using a validated clinical tool, as well as the hemostatic interventions and clinical outcomes associated with bleeding.

Design: Prospective observational cohort study.

Setting: Tertiary pediatric critical care unit

Patients: All consecutive patients (1 month to 18 years of age) admitted to a tertiary pediatric critical care unit

Measurements and Main Results: Bleeding events were categorized as minimal, moderate, severe, or fatal, according to the Bleeding Assessment Scale in Critically Ill Children. We collected demographics and severity at admission, as evaluated by the Pediatric Index of Mortality. We used regression models to compare the severity of bleeding with outcomes adjusting for age, surgery, and severity. Over 12 months, 902 critically ill patients were enrolled. The median age was 64 months (IQR 17; 159), the median admission predicted risk of mortality was 0.5% (IQR 0.2; 1.4), and 24% were post-surgical. Eighteen percent of patients experienced at least one bleeding event. The highest severity of bleeding was minimal for 7.9% of patients, moderate for 5.8%, severe for 3.8%, and fatal for 0.1%. Adjusting for age, severity at admission, medical diagnosis, type of surgery, and duration of surgery, bleeding severity was independently associated with fewer ventilator-free days (p < 0.001) and fewer PICU-free days (p < 0.001). Adjusting for the same variables, bleeding severity was independently associated with an increased risk of mortality (adjusted odds ratio for each bleeding category 2.4, 95% CI 1.5; 3.7, p < 0.001).

Conclusion: Our data indicate bleeding occurs in nearly one-fifth of all critically ill children, and that higher severity of bleeding was independently associated with worse clinical outcome. Further multicenter studies are required to better understand the impact of bleeding in critically ill children.
Novel Mechanism for an Old Drug: Phenazopyridine is a Kinase Inhibitor Affecting Autophagy and Cellular Differentiation
Authors: Preynat-Seauve Olivier , Nguyen Evelyne Bao-Vi , Westermaier Yvonne , Héritier Margaux , Tardy Sébastien , Cambet Yves , Feyeux Maxime , Caillon Aurélie , Scapozza Leonardo , Krause Karl-Heinz
Published: August 2021

Phenazopyridine is a widely used drug against urinary tract pain. The compound has also been shown to enhance neural differentiation of pluripotent stem cells. However, its mechanism of action is not understood. Based on its chemical structure, we hypothesized that phenazopyridine could be a kinase inhibitor. Phenazopyridine was investigated in the following experimental systems: 1) activity of kinases in pluripotent stem cells; 2) binding to recombinant kinases, and 3) functional impact on pluripotent stem cells. Upon addition to pluripotent stem cells, phenazopyridine induced changes in kinase activities, particularly involving Mitogen-Activated Protein Kinases, Cyclin-Dependent Kinases, and AKT pathway kinases. To identify the primary targets of phenazopyridine, we screened its interactions with 401 human kinases. Dose-inhibition curves showed that three of these kinases interacted with phenazopyridine with sub-micromolar binding affinities: cyclin-G-associated kinase, and the two phosphatidylinositol kinases PI4KB and PIP4K2C, the latter being known for participating in pain induction. Docking revealed that phenazopyridine forms strong H-bonds with the hinge region of the ATP-binding pocket of these kinases. As previous studies suggested increased autophagy upon inhibition of the phosphatidyl-inositol/AKT pathway, we also investigated the impact of phenazopyridine on this pathway and found an upregulation. In conclusion, our study demonstrates for the first time that phenazopyridine is a kinase inhibitor, impacting notably phosphatidylinositol kinases involved in nociception.
High-Throughput Drug Screening System Based on Human Induced Pluripotent Stem Cell-Derived Atrial Myocytes ∼ A Novel Platform to Detect Cardiac Toxicity for Atrial Arrhythmias
Authors: Honda Yayoi , Li Jun , Hino Aya , Tsujimoto Shinji , Lee Jong-Kook
Published: August 2021

Evaluation of proarrhythmic properties is critical for drug discovery. In particular, QT prolongation in electrocardiograms has been utilized as a surrogate marker in many evaluation systems to assess the risk of torsade de pointes and lethal ventricular arrhythmia. Recently, new evaluation systems based on human iPS cell-derived cardiomyocytes have been established. On the other hand, in clinical situations, it has been reported that the incidence of atrial arrhythmias such as atrial fibrillation has been increasing every year, with the prediction of a persistent increase in the near future. As to the increased incidence of atrial arrhythmias, in addition to the increased population of geriatric patients, a wide variety of drug treatments may be related, as an experimental method to detect drug-induced atrial arrhythmia has not been established so far. In the present study, we characterized the atrial-like cardiomyocytes derived from human induced pluripotent stem cells and examined their potential for the evaluation of drug-induced atrial arrhythmia. Atrial-like cardiomyocytes were induced by adding retinoic acid (RA) during the process of myocardial differentiation, and their characteristics were compared to those of RA-free cardiomyocytes. Using gene expression and membrane potential analysis, it was confirmed that the cells with or without RA treatment have atrial or ventricular like cardiomyocytes, respectively. Using the ultra-rapid activating delayed rectifier potassium current (IKur) channel inhibitor, which is specific to atrial cardiomyocytes, Pulse width duration (PWD) 30cF prolongation was confirmed only in atrial-like cardiomyocytes. In addition, ventricular like cardiomyocytes exhibited an early after depolarization by treatment with rapidly activating delayed rectifier potassium current (IKr) channel inhibitor, which induces ventricular arrhythmia in clinical situations. Here, we have established a high-throughput drug evaluation system using human iPS cell-derived atrial-like cardiomyocytes. Based on the obtained data, the system might be a valuable platform to detect potential risks for drug-induced atrial arrhythmias.
Plastome Evolution in the Hyperdiverse Genus Euphorbia (Euphorbiaceae) Using Phylogenomic and Comparative Analyses: Large-Scale Expansion and Contraction of the Inverted Repeat Region
Authors: Wei Neng , Pérez-Escobar Oscar A. , Musili Paul M. , Huang Wei-Chang , Yang Jun-Bo , Hu Ai-Qun , Hu Guang-Wan , Grace Olwen M. , Wang Qing-Feng
Published: August 2021

Withc.2,000 species,Euphorbiais one of the largest angiosperm genera, yet a lack of chloroplast genome (plastome) resources impedes a better understanding of its evolution. In this study, we assembled and annotated 28 plastomes from Euphorbiaceae, of which 15 were newly sequenced. Phylogenomic and comparative analyses of 22 plastome sequences from all four recognized subgenera withinEuphorbiarevealed that plastome length inEuphorbiais labile, presenting a range of variationc.42 kb. Large-scale expansions of the inverted repeat (IR) region were identified, and at the extreme opposite, the near-complete loss of the IR region (with only 355 bp left) was detected for the first time in Euphorbiaceae. Other structural variations, including gene inversion and duplication, and gene loss/pseudogenization, were also observed. We screened the most promising molecular markers from both intergenic and coding regions for phylogeny-based utilities, and estimated maximum likelihood and Bayesian phylogenies from four datasets including whole plastome sequences. The monophyly ofEuphorbiais supported, and its four subgenera are recovered in a successive sister relationship. Our study constitutes the first comprehensive investigation on the plastome structural variation inEuphorbiaand it provides resources for phylogenetic research in the genus, facilitating further studies on its taxonomy, evolution, and conservation.
Detailed Emotional Profile of Secondary Education Students Toward Learning Physics and Chemistry
Authors: Dávila-Acedo María Antonia , Airado-Rodríguez Diego , Cañada-Cañada Florentina , Sánchez-Martín Jesús
Published: August 2021

The present research arises from the need to identify the emotions that K-7 to K-10 students experience toward the learning of Physics and Chemistry, since it is a fact that there is a decrease in the number of students choosing itineraries related to Science. Different blocks of contents have been considered in each subject in order to identify emotions toward each one of them. The considered sample consisted of 149 K-8 students, 152 K-9 students and 130 K-10 students from several middle and high schools in Badajoz (Spain) during the 2014–2015 school year. Students experienced more positive emotions toward the content of Chemistry than toward those of Physics. A decrease was detected in the mean frequency of positive emotions such as joy, fun, and tranquility from K-8 to K-10, as well as an increase in negative emotions such as boredom, anxiety, disgust, fear, nervousness, worry, and sadness. It has also been found that positive emotions toward Chemistry contents are mainly related to teachers’ methods and attitudes, while negative emotions toward Physics contents are related to the exclusive use of the textbook, solving Physics problems, or giving oral presentations of the topics in class.
Are Narrow Focus Exhaustivity Inferences Bayesian Inferences?
Authors: Schreiber Alexander , Onea Edgar
Published: August 2021

In successful communication, the literal meaning of linguistic utterances is often enriched by pragmatic inferences. Part of the pragmatic reasoning underlying such inferences has been successfully modeled as Bayesian goal recognition in the Rational Speech Act (RSA) framework. In this paper, we try to model the interpretation of question-answer sequences with narrow focus in the answer in the RSA framework, thereby exploring the effects of domain size and prior probabilities on interpretation. Should narrow focus exhaustivity inferences be actually based on Bayesian inference involving prior probabilities of states, RSA models should predict a dependency of exhaustivity on these factors. We present experimental data that suggest that interlocutors do not act according to the predictions of the RSA model and that exhaustivity is in fact approximately constant across different domain sizes and priors. The results constitute a conceptual challenge for Bayesian accounts of the underlying pragmatic inferences.
Electrophysiological Correlates of Character Transposition in the Left and Right Visual Fields
Authors: Zhang Er-Hu , Lai Xue-Xian , Li Defeng , Lei Victoria Lai Cheng , Chen Yiqiang , Cao Hong-Wen
Published: August 2021

This study examined the brain activity elicited by the hemispheric asymmetries and morpheme transposition of two-character Chinese words (canonical and transposed word) and pseudowords using event-related potentials (ERPs) with a dual-target rapid serial visual presentation (RSVP) task. Electrophysiological results showed facilitation effects for canonical words with centrally presented visual field (CVF) and right visual field (RVF) presentations but not with left visual field (LVF) presentations, as reflected by less negative N400 amplitudes. Moreover, more positive late positive component (LPC) amplitudes were observed for both canonical words and transposed words irrespective of the visual fields. More importantly, transposed words elicited a more negative N400 amplitude and a less positive LPC amplitude compared with the amplitudes elicited by canonical words for CVF and RVF presentations. For LVF presentations, transposed words elicited a less negative N250 amplitude compared with canonical words, and there was no significant difference between canonical words and transposed words in the N400 effect. Taken together, we concluded that character transposition facilitated the mapping of whole-word orthographic representation to semantic information in the LVF, as reflected by the N250 component, and such morpheme transposition influenced whole-word semantic processing in CVF and RVF presentations, as reflected by N400 and LPC components.
Anatomical Study of the Hepatic Veins in Segment 4 of the Liver Using Three-Dimensional Visualization
Authors: Zhang Jun , Guo Xiaochao , Qiao Qilu , Zhao Jianxun , Wang Xin
Published: August 2021

Objective: The current study aimed to examine the anatomical structure of the hepatic vein of segment IV liver (S4) of the liver using three-dimensional (3D) visualization technology in order to explore the surgical value of the middle hepatic vein (MHV) manipulation and highlight the importance of current research in hepatic surgery.

Methods: Between January 2014 and December 2019, 52 patients with abdominal diseases(not including hepatic disease) were selected for multiphasic computed tomography-enhanced scans of the upper abdomen. A 3D visualization system was utilized to display the structural details of the hepatic veins in S4 of their livers. Couinaud's eight-segment classification system was used to denote the liver' sections.

Results: The constructed 3D model clearly displayed vascular morphological characteristics and their location in the liver, hepatic artery and vein system, and portal vein system. Of the 52 patients, 43 had an umbilical fissure vein (UFV) (82.7%), 19 had an accessory S4 liver vein (36.5%), 16 had both a UFV (30.8%) and an accessory S4 liver vein, and 6 had neither (11.5%). A total of 79% of the patients with a UFV and 74.2% of those with an accessory S4 liver vein had venous blood returning into the left hepatic vein.

Conclusion: 3D visualization technology was used to determine hepatic venous return of S4 hepatic veins and was found to improve the safety of evaluation in hepatic surgery.
In Vitro Antimicrobial Susceptibilities of Francisella tularensis subsp. holarctica Isolates from Tularemia Outbreaks That Occurred from the End of the 20th Century to the 2020s in Spain
Authors: Martínez-Martínez Sonia , Rodríguez-Ferri Elías-Fernando , Rodríguez-Lázaro David , Hernández Marta , Gómez-Campillo José-Ignacio , Martínez-Nistal María del Carmen , Fernández-Natal María-Isabel , García-Iglesias María-José , Mínguez-González Olga , Gutiérrez-Martín César-Bernardo
Published: August 2021

A collection of 177 Francisella tularensis subsp. holarctica clinical isolates (29 from humans and 148 from animals, mainly hares and voles) was gathered from diverse tularemia outbreaks in the Castilla y León region (northwestern Spain) that occurred from the end of the 20th century to the 2020s. Along with four F. tularensis subsp. holarctica reference strains, all of these clinical isolates were tested using a broth microdilution method to determine their susceptibility to 22 antimicrobial agents, including β-lactams, aminoglycosides and one member each of the tetracycline, glycylcycline, quinolone and sulphonamide classes. Many multi-resistance profiles were found among the tested isolates, but especially among those of human origin (all but two isolates showed resistance to at least 13 of 18 antimicrobial agents). Even so, all human isolates were susceptible to gentamicin and tobramycin, while more than 96% of animal isolates were susceptible to these two aminoglycosides. Ciprofloxacin showed activity against more than 92% of animal and human isolates. However, almost 21% of human isolates were resistant to tetracycline, and more than 65% were resistant to tigecycline. Finally, a quite similar activity to other F. tularensis subsp. holarctica isolates collected 20 years earlier in Spain was observed.