bluuebunny/crossref_metadata_2025 huggingface dataset:
TEACHING ABOUT ROLLING MOTION: EXPLORING THE EFFECTIVENESS OF AN EXTREME CASE REASONING APPROACH
Authors: Vidak Andrej , Erceg Nataša , Hasović Elvedin , Odžak Senad , Mešić Vanes
Published: February 2021

Earlier research has shown that students have tremendous difficulties with understanding certain aspects of rolling without slipping, such as the zero-velocity at the contact point and plausibility of application of the law of conservation of mechanical energy despite action of the friction force. The aim of this research was to explore whether using analogies and reasoning about extreme cases can facilitate conceptualization of the above-mentioned phenomena. A pre-test – post-test quasi-experiment has been conducted, with 93 students in the control group (CG) and 91 students in the experimental group (EG). Whereas control group students received conventional teaching, in the experimental group rolling of a cylinder has been considered as a special case of a tumbling prism for which the number of prism surfaces tended to infinity. The results of analysis of covariance showed that students from the experimental group significantly outperformed their peers from the control group on the Rolling Motion Concept Test (RMCT). Between-group differences were greater on test items that required higher level of cognitive transfer. This research suggests that using analogies and extreme case reasoning can facilitate comprehension of certain seemingly counterintuitive aspects of rolling motion.
Keywords: analogy-based teaching, energy conservation, extreme case reasoning, misconceptions, rolling motion.
PENGGUNAAN CLIP (KEPINGAN) PADA MATERI PENJUMLAHAN DAN MENGURANGAN BILANGAN BULAT DALAM MENINGKATKAN AKTIVITAS DAN HASIL BELAJAR MATEMATIKA
Authors: Wirahmad Ika Ika , Kurniawati Kurniawati
Published: February 2021

Penelitian ini adalah penelitian tindakan kelas yang bertujuan untuk meningkatkan aktivitas dan hasil belajar matematika dengan penggunaan clip(kepingan). Subjek penelitian ini adalah seluruh siswa kelas VIIA SMP Negeri 2 Donggo yang berjumlah 28 orang. Jenis penelitian ini adalah jenis penelitian tindakan kelas dengan Rancangan penelitian yang mengacu pada model penelitian yang dikemukakan oleh Kemmis dan Mc. Taggart (Arikunto, 2008: 16) yang terdiri atas 4 tahapan yaitu (1) perencanaan, (2) pelaksanaan tindakan, (3) observasi/Evaluasi dan (4) refleksi. Pengambilan data di lakukan dengan menggunakan tes hasil belajar dan hasil observasi. Data yang terkumpul di analisis dengan menggunakan analisis kualitatif dan analisis kuantitatif dengan indikator keberhasilannya dikatakan meningkat jika hasil belajar memenuhi KKM minimal ≥65 dan ketuntasan Klasikal ≥85%. Hasil penelitian menunjukan bahwa terdapat peningkatan perolehan skor rata-rata 60 berkategori rendah pada siklus I dan ketuntasan klaksikal mencapai 67,85 meningkat menjadi 85% pada siklus II dengan  perolehan skor rata-rata 70 paling rendah  pada siklus II dengan ketuntasa klaksikal mencapai  92,85. Ketuntasan klaksikal yang di tetapkan oleh depertemen pendidkan nasional yaitu 85%. Pada siklus I belum memenuhi ketuntasan klaksikal dan terjadi peningkatan pada siklus II sehingga ketuntasan klaksikal terpenuhi. Dapat di simpulkan bahwa penggunaan clip (kepingan) pada materi penjumlahan dan pengurangan bilangan bulat dapat meningkatkan aktivitas dah hasil belajar matematika siswa kelas VIIA  SMP Negeri 2 Donggo Tahun Pelajaran 2017/2018
THE FACTORS AFFECTING THE PROBLEMATIC INTERNET USAGE AND THE SOCIALIZATION LEVEL AMONG HEARING-IMPAIRED ADOLESCENTS
Authors: AKTAS KUCUKTAS Tugce , BÜLBÜL Tülay , ÖZEN Betül
Published: February 2021

This descriptive study was conducted to identify the factors affecting problematic internet usage and the socialization level among hearing-impaired adolescents. The study has been done with one hundred twenty six (97 males and 29 females) hearing-impaired students, aged between 14 and 21. The data were collected using Turkish Sign Language, by the researcher who received Sign Language Interpretation Training. ‘The Problematic Internet Usage Scale’ and ‘Socialization Level Scale’ were used as assessment instruments. The results showed that level of the problematic internet usage and the socialization of the participants were at a moderate level with mean scores of 83.8±19.8 on the Problematic Internet Usage Scale and 31.9±8.9 on the Socialization Level Scale. As hearing-impaired adolescents are encouraged to use more internet in order to increase their socialization level, risks as; problematic internet use, keeping young people away from society, and social exclusion should be taken into consideration.
YOĞUN BAKIMDA HASTASI OLAN AİLE BİREYLERİNİN GEREKSİNİMLERİNİN KARŞILANMASI İLE MEMNUNİYET DÜZEYLERİ ARASINDAKİ İLİŞKİ
Authors: ÖZTÜRK Esra , CERİT Birgül
Published: February 2021

Araştırma, yoğun bakımda hastası olan aile bireylerinin gereksinimlerinin karşılanması ile memnuniyet düzeyleri arasındaki ilişkiyi belirlemek amacıyla gerçekleştirilmiştir. Araştırma ilişkisel tarama modeli kullanılarak tanımlayıcı tipte yapılmıştır. Veriler çalışmaya katılmaya gönüllü olan 296 hasta yakınından elde edilmiştir. Araştırma verileri Kişisel Bilgi Formu, Karşılanan Gereksinimler Envanteri ve Yoğun Bakım Memnuniyet Ölçeği kullanılarak toplanmıştır. Verilerin analizinde, tanımlayıcı istatistikler, Pearson’s korelasyon analizi ve çoklu regresyon analizi kullanılmıştır. Hasta yakınlarının yoğun bakım memnuniyetini, güven, bilgi ve yakınlık gereksinimlerinin karşılanması alt boyutlarının yordadığı, bilgi memnuniyetini, güven ve bilgi gereksinimlerinin karşılanması alt boyutlarının yordadığı, karar verme memnuniyetini ise güven ve bilgi gereksinimlerinin karşılanması alt boyutlarının yordadığı belirlenmiştir. Araştırma sonucu, hasta yakınlarının gereksinimlerinin karşılanmasının yoğun bakım memnuniyetini yordadığını göstermektedir. Dolayısıyla yoğun bakımda hastası olan aile bireylerinin gereksinimlerinin karşılanmasına yönelik düzenlemelerin yapılması yoğun bakım memnuniyetine olumlu yansıyabilir.
REMARKS ON THE ARITHMETICAL FUNCTION $a_{p}(n)$
Authors: ADEM Abdu Awel
Published: February 2021

In this paper,for any arbitrary two prime numbers $p$ and $q$ the relationship between the corresponding arithmetic functions $(a_p(n))$ and $(a_q(n))$ are investigated.~Furthermore, the general formula for statistical density of all sets on which the two arithmetic functions have the same value also established.
Periodontal Health Status and Treatment Needs of Visually Impaired Student Attending “Extraordinary School in Jember Regency
Authors: Dewi Ni Luh Putu Diah Laksmi , Budirahardjo Roedy , Sulistiyani Sulistiyani
Published: February 2021

Visually impaired people usually have limitation to do their activities in maintaining oral health including the periodontal tissue. This condition leads to inadequate efforts to maintain oral health that can cause various diseases, one of which is periodontal disease. Periodontal disease will gradually develop become severe, and therefore it is necessary to take proper care in handling it. The aim of this study was to determine the periodontal health status and the level of periodontal disease care needs of blind students in SLB Jember Regency. This research was a descriptive observational study with a cross sectional approach. The research sample of 20 people were obtained using the total sampling method. The health status of the periodontal tissue and the need for treatment are examined using CPITN. Subsequent research data are grouped and distributed based on gender and age. The result showed that the most common periodontal tissue health status was score two with clinical features of subgingival calculus or supragingival calculus with a total of 50%, while the most needed treatment needs were improvement in oral hygiene accompanied by 85% professional scaling.

Keywords: periodontal health status; periodontal treatment needs; CPITN; visually impaired
Nursing Profession, Caring and Discipline
Authors: MARTININGSIH Wiwin , Winarni Sri , Alvarado Angelito Estoesta
Published: February 2021

Nursing indeed is the art of caring and selfless giving of time, effort, love, and care. The patient is the center of nursing, and it is their responsibility to make sure they are receiving the right care that they can receive. There are so many ways to show caring that the possibilities never end. The purpose of this study was to generate knowledge which informed caring in nursing as a discipline within applying attributes of caring. Mayeroff and Roach’s attributes guided the concept analysis within the theoretical perspective of caring. The emerging themes are: Caring as the central focus of Nursing, Caring as a Profession and Discipline, Our perspective of caring, Our idea of a caring nurse, and How does Nurse care for others. Caring is everything and nurse should applying attributes of caring in the fields of nursing. Caring is not only able to care for the client, but also how the client is satisfied and the nurse always continues to improve knowledge and be innovative and creative.

Keywords: nursing profession; caring; discipline
Effect of Electrical Stimulation Conditions on Neural Stem Cells Differentiation on Cross-Linked PEDOT:PSS Films
Authors: Sordini Laura , Garrudo Fábio F. F. , Rodrigues Carlos A. V. , Linhardt Robert J. , Cabral Joaquim M. S. , Ferreira Frederico Castelo , Morgado Jorge
Published: February 2021

The ability to culture and differentiate neural stem cells (NSCs) to generate functional neural populations is attracting increasing attention due to its potential to enable cell-therapies to treat neurodegenerative diseases. Recent studies have shown that electrical stimulation improves neuronal differentiation of stem cells populations, highlighting the importance of the development of electroconductive biocompatible materials for NSC culture and differentiation for tissue engineering and regenerative medicine. Here, we report the use of the conjugated polymer poly(3,4-ethylenedioxythiophene) doped with polystyrene sulfonate (PEDOT:PSS CLEVIOS P AI 4083) for the manufacture of conductive substrates. Two different protocols, using different cross-linkers (3-glycidyloxypropyl)trimethoxysilane (GOPS) and divinyl sulfone (DVS) were tested to enhance their stability in aqueous environments. Both cross-linking treatments influence PEDOT:PSS properties, namely conductivity and contact angle. However, only GOPS-cross-linked films demonstrated to maintain conductivity and thickness during their incubation in water for 15 days. GOPS-cross-linked films were used to culture ReNcell-VM under different electrical stimulation conditions (AC, DC, and pulsed DC electrical fields). The polymeric substrate exhibits adequate physicochemical properties to promote cell adhesion and growth, as assessed by Alamar Blue® assay, both with and without the application of electric fields. NSCs differentiation was studied by immunofluorescence and quantitative real-time polymerase chain reaction. This study demonstrates that the pulsed DC stimulation (1 V/cm for 12 days), is the most efficient at enhancing the differentiation of NSCs into neurons.
Bornyl Diphosphate Synthase From Cinnamomum burmanni and Its Application for (+)-Borneol Biosynthesis in Yeast
Authors: Ma Rui , Su Ping , Guo Juan , Jin Baolong , Ma Qing , Zhang Haiyan , Chen Lingli , Mao Liuying , Tian Mei , Lai Changjiangsheng , Tang Jinfu , Cui Guanghong , Huang Luqi
Published: February 2021

(+)-Borneol is a desirable monoterpenoid with effective anti-inflammatory and analgesic effects that is known as soft gold. (+)-bornyl diphosphate synthase is the key enzyme in the (+)-borneol biosynthesis pathway. Despite several reported (+)-bornyl diphosphate synthase genes, relatively low (+)-borneol production hinders the attempts to synthesize it using microbial fermentation. Here, we identified the highly specific (+)-bornyl diphosphate synthase CbTPS1 from Cinnamomum burmanni. An in vitro assay showed that (+)-borneol was the main product of CbTPS1 (88.70% of the total products), and the Km value was 5.11 ± 1.70 μM with a kcat value of 0.01 s–1. Further, we reconstituted the (+)-borneol biosynthetic pathway in Saccharomyces cerevisiae. After tailored truncation and adding Kozak sequences, the (+)-borneol yield was improved by 96.33-fold to 2.89 mg⋅L–1 compared with the initial strain in shake flasks. This work is the first reported attempt to produce (+)-borneol by microbial fermentation. It lays a foundation for further pathway reconstruction and metabolic engineering production of this valuable natural monoterpenoid.
Modeling of the Beam-To-Column Dowel Connection for a Single-Story RC Precast Building
Authors: Magliulo Gennaro , Di Salvatore Chiara , Ercolino Marianna
Published: February 2021

As widely known, connections performance under seismic loads can significantly affect the structural response of RC precast buildings. Within the scientific community, an increasing number of studies has been carried out on this topic, in the light of the recent earthquake aftermaths all over Europe. Indeed, connections turned out to be the weakest part of the precast structures and their failure often provoked the global collapse of the whole building. The present study aims at assessing the seismic behavior of a single-story RC precast building in terms of global collapse implementing two different models of the beam-to-column connection, a simplified and a refined one. A lumped plasticity approach is used to simulate the structural nonlinear behavior at the column base. In order to assess the refined connection model, a preliminary scheme with an isolated single dowel is validated by comparing the pushover outcomes with experimental results from literature. Nonlinear static and dynamic analyses are performed on two models of a 3D single-story RC precast building, one implementing the simple beam-to-column connection model and the other one implementing the refined mode. The comparison clearly shows that the differences are negligible if the global collapse limit state is considered.
Mutant p53 Gain-of-Function: Role in Cancer Development, Progression, and Therapeutic Approaches
Authors: Alvarado-Ortiz Eduardo , de la Cruz-López Karen Griselda , Becerril-Rico Jared , Sarabia-Sánchez Miguel Angel , Ortiz-Sánchez Elizabeth , García-Carrancá Alejandro
Published: February 2021

Frequent p53 mutations (mutp53) not only abolish tumor suppressor capacities but confer various gain-of-function (GOF) activities that impacts molecules and pathways now regarded as central for tumor development and progression. Although the complete impact of GOF is still far from being fully understood, the effects on proliferation, migration, metabolic reprogramming, and immune evasion, among others, certainly constitute major driving forces for human tumors harboring them. In this review we discuss major molecular mechanisms driven by mutp53 GOF. We present novel mechanistic insights on their effects over key functional molecules and processes involved in cancer. We analyze new mechanistic insights impacting processes such as immune system evasion, metabolic reprogramming, and stemness. In particular, the increased lipogenic activity through the mevalonate pathway (MVA) and the alteration of metabolic homeostasis due to interactions between mutp53 and AMP-activated protein kinase (AMPK) and Sterol regulatory element-binding protein 1 (SREBP1) that impact anabolic pathways and favor metabolic reprograming. We address, in detail, the impact of mutp53 over metabolic reprogramming and the Warburg effect observed in cancer cells as a consequence, not only of loss-of-function of p53, but rather as an effect of GOF that is crucial for the imbalance between glycolysis and oxidative phosphorylation. Additionally, transcriptional activation of new targets, resulting from interaction of mutp53 with NF-kB, HIF-1α, or SREBP1, are presented and discussed. Finally, we discuss perspectives for targeting molecules and pathways involved in chemo-resistance of tumor cells resulting from mutp53 GOF. We discuss and stress the fact that the status of p53 currently constitutes one of the most relevant criteria to understand the role of autophagy as a survival mechanism in cancer, and propose new therapeutic approaches that could promote the reduction of GOF effects exercised by mutp53 in cancer.
Age-Related Gene Alteration in Naïve and Memory T cells Using Precise Age-Tracking Model
Authors: Yang Xiaofeng , Wang Xin , Lei Lei , Sun Lina , Jiao Anjun , Zhu Kun , Xie Tao , Liu Haiyan , Zhang Xingzhe , Su Yanhong , Zhang Cangang , Shi Lin , Zhang Dan , Zheng Huiqiang , Zhang Jiahui , Liu Xiaobin , Wang Xin , Zhou Xiaobo , Sun Chenming , Zhang Baojun
Published: February 2021

In aged individuals, age-related changes in immune cells, especially T cell deficiency, are associated with an increased incidence of infection, tumor, and autoimmune disease, as well as an impaired response to vaccination. However, the features of gene expression levels in aged T cells are still unknown. Our previous study successfully tracked aged T cells generated from one wave of developing thymocytes of young age by a lineage-specific and inducible Cre-controlled reporter (TCRδCreERR26ZsGreen mouse strain). In this study, we utilized this model and genome-wide transcriptomic analysis to examine changes in gene expression in aged naïve and memory T cell populations during the aging process. We identified profound gene alterations in aged CD4 and CD8 T cells. Both aged CD4+ and CD8+ naïve T cells showed significantly decreased organelle function. Importantly, genes associated with lymphocyte activation and function demonstrated a significant increase in aged memory T cells, accompanied by upregulation of immunosuppressive markers and immune checkpoints, revealing an abnormal T cell function in aged cells. Furthermore, aging significantly affects T cell survival and death signaling. While aged CD4 memory T cells exhibited pro-apoptotic gene signatures, aged CD8 memory T cells expressed anti-apoptotic genes. Thus, the transcriptional analysis of gene expression and signaling pathways in aged T cell subsets shed light on our understanding of altered immune function with aging, which will have great potential for clinical interventions for older adults.
Role of Non-coding RNAs on the Radiotherapy Sensitivity and Resistance of Head and Neck Cancer: From Basic Research to Clinical Application
Authors: Zhang Xixia , Yang Jing
Published: February 2021

Head and neck cancers (HNCs) rank as the sixth common and the seventh leading cause of cancer-related death worldwide, with an estimated incidence of 600,000 cases and 40–50% mortality rate every year. Radiotherapy is a common local therapeutic modality for HNC mainly through the function of ionizing radiation, with approximately 60% of patients treated with radiotherapy or chemoradiotherapy. Although radiotherapy is more advanced and widely used in clinical practice, the 5-year overall survival rates of locally advanced HNCs are still less than 40%. HNC cell resistance to radiotherapy remains one of the major challenges to improve the overall survival in HNC patients. Non-coding RNAs (ncRNAs) are newly discovered functional small RNA molecules that are different from messenger RNAs, which can be translated into a protein. Many previous studies have reported the dysregulation and function of ncRNAs in HNC. Importantly, researchers reported that several ncRNAs were also dysregulated in radiotherapy-sensitive or radiotherapy-resistant HNC tissues compared with the normal cancer tissues. They found that ectopically elevating or knocking down expression of some ncRNAs could significantly influence the response of HNC cancer cells to radiotherapy, indicating that ncRNAs could regulate the sensitivity of cancer cells to radiotherapy. The implying mechanism for ncRNAs in regulating radiotherapy sensitivity may be due to its roles on affecting DNA damage sensation, inducing cell cycle arrest, regulating DNA damage repair, modulating cell apoptosis, etc. Additionally, clinical studies reported thatin situncRNA expression in HNC tissues may predict the response of radiotherapy, and circulating ncRNA from body liquid serves as minimally invasive therapy-responsive and prognostic biomarkers in HNC. In this review, we aimed to summarize the current function and mechanism of ncRNAs in regulating the sensitivity of HNC cancer cells to radiotherapy and comprehensively described the state of the art on the role of ncRNAs in the prognosis prediction, therapy monitoring, and prediction of response to radiotherapy in HNC.
FOntCell: Fusion of Ontologies of Cells
Authors: Cabau-Laporta Javier , Ascensión Alex M. , Arrospide-Elgarresta Mikel , Gerovska Daniela , Araúzo-Bravo Marcos J.
Published: February 2021

High-throughput cell-data technologies such as single-cell RNA-seq create a demand for algorithms for automatic cell classification and characterization. There exist several cell classification ontologies with complementary information. However, one needs to merge them to synergistically combine their information. The main difficulty in merging is to match the ontologies since they use different naming conventions. Therefore, we developed an algorithm that merges ontologies by integrating the name matching between class label names with the structure mapping between the ontology elements based on graph convolution. Since the structure mapping is a time consuming process, we designed two methods to perform the graph convolution: vectorial structure matching and constraint-based structure matching. To perform the vectorial structure matching, we designed a general method to calculate the similarities between vectors of different lengths for different metrics. Additionally, we adapted the slower Blondel method to work for structure matching. We implemented our algorithms into FOntCell, a software module in Python for efficient automatic parallel-computed merging/fusion of ontologies in the same or similar knowledge domains. FOntCell can unify dispersed knowledge from one domain into a unique ontology in OWL format and iteratively reuse it to continuously adapt ontologies with new data endlessly produced by data-driven classification methods, such as of the Human Cell Atlas. To navigate easily across the merged ontologies, it generates HTML files with tabulated and graphic summaries, and interactive circular Directed Acyclic Graphs. We used FOntCell to merge the CELDA, LifeMap and LungMAP Human Anatomy cell ontologies into a comprehensive cell ontology. We compared FOntCell with tools used for the alignment of mouse and human anatomy ontologies task proposed by the Ontology Alignment Evaluation Initiative (OAEI) and found that the Fβ alignment accuracies of FOntCell are above the geometric mean of the other tools; more importantly, it outperforms significantly the best OAEI tools in cell ontology alignment in terms of Fβ alignment accuracies.
The Lack of SNARE Protein Homolog Syn8 Influences Biofilm Formation of Candida glabrata
Authors: Chen Xinyue , Iwatani Shun , Kitamoto Toshitaka , Chibana Hiroji , Kajiwara Susumu
Published: February 2021

Biofilm formation of Candida species is considered to be a pathogenic factor of host infection. Since biofilm formation of Candida glabrata has not been as well studied as that of Candida albicans, we performed genetic screening of C. glabrata, and three candidate genes associated with biofilm formation were identified. Candida glabrata SYN8 (CAGL0H06325g) was selected as the most induced gene in biofilm cells for further research. Our results indicated that the syn8Δ mutant was defective not only in biofilm metabolic activity but also in biofilm morphological structure and biomass. Deletion of SYN8 seemed to have no effect on extracellular matrix production, but it led to a notable decrease in adhesion ability during biofilm formation, which may be linked to the repression of two adhesin genes, EPA10 and EPA22. Furthermore, hypersensitivity to hygromycin B and various ions in addition to the abnormal vacuolar morphology in the syn8Δ mutant suggested that active vacuolar function is required for biofilm formation of C. glabrata. These findings enhance our understanding of biofilm formation in this fungus and provide information for the development of future clinical treatments.
BMPER Ameliorates Renal Fibrosis by Inhibiting Tubular Dedifferentiation and Fibroblast Activation
Authors: Xie Ting , Xia Zunen , Wang Wei , Zhou Xiangjun , Xu Changgeng
Published: February 2021

Tubulointerstitial fibrosis is both a pathological manifestation of chronic kidney disease and a driving force for the progression of kidney disease. A previous study has shown that bone morphogenetic protein-binding endothelial cell precursor-derived regulator (BMPER) is involved in lung fibrogenesis. However, the role of BMPER in renal fibrosis remains unknown. In the present study, the expression of BMPER was examined by real-time PCR, Western blot and immunohistochemical staining. The in vitro effects of BMPER on tubular dedifferentiation and fibroblast activation were analyzed in cultured HK-2 and NRK-49F cells. The in vivo effects of BMPER were dissected in unilateral ureteral obstruction (UUO) mice by delivery of BMPER gene via systemic administration of plasmid vector. We reported that the expression of BMPER decreased in the kidneys of UUO mice and HK-2 cells. TGF-β1 increased inhibitor of differentiation-1 (Id-1) and induced epithelial mesenchymal transition in HK-2 cells, and knockdown of BMPER aggravated Id-1 up-regulation, E-cadherin loss, and tubular dedifferentiation. On the contrary, exogenous BMPER inhibited Id-1 up-regulation, prevented E-cadherin loss and tubular dedifferentiation after TGF-β1 exposure. In addition, exogenous BMPER suppressed fibroblast activation by hindering Erk1/2 phosphorylation. Knockdown of low-density lipoprotein receptor-related protein 1 abolished the inhibitory effect of BMPER on Erk1/2 phosphorylation and fibroblast activation. Moreover, delivery of BMPER gene improved renal tubular damage and interstitial fibrosis in UUO mice. Therefore, BMPER inhibits TGF-β1-induced tubular dedifferentiation and fibroblast activation and may hold therapeutic potential for tubulointerstitial fibrosis.
NADPH and Mito-Apocynin Treatment Protects Against KA-Induced Excitotoxic Injury Through Autophagy Pathway
Authors: Liu Na , Lin Miao-Miao , Huang Si-Si , Liu Zi-Qi , Wu Jun-Chao , Liang Zhong-Qin , Qin Zheng-Hong , Wang Yan
Published: February 2021

Aim

Previous research recognizes that NADPH can produce reduced glutathione (GSH) as a coenzyme and produce ROS as a substrate of NADPH oxidase (NOX). Besides, excessive activation of glutamate receptors results in mitochondrial impairment. The study aims at spelling out the effects of NADPH and Mito-apocynin, a NOX inhibitor which specifically targets the mitochondria, on the excitotoxicity induced by Kainic acid (KA) and its mechanism.

Methods

Thein vivoneuronal excitotoxicity model was constructed by stereotypically injecting KA into the unilateral striatum of mice. Administrated NADPH (i.v, intravenous) 30 min prior and Mito-apocynin (i.g, intragastric) 1 day prior, respectively, then kept administrating daily until mice were sacrificed 14 days later. Nissl staining measured the lesion of striatum and survival status of neurons. Cylinder test of forelimb asymmetry and the adhesive removal test reflected the behavioral deficit caused by neural dysfunction. Determined Total superoxide dismutase (T-SOD), malondialdehyde (MDA), and GSH indicated oxidative stress. Western blot presented the expression levels of LC3-II/LC3-I, SQSTM1/p62, TIGAR, and NOX4. Assessed oxygen consumption rate using High-Resolution Respirometry.In vitro, the MitoSOX Indicator reflected superoxide released by neuron mitochondria. JC-1 and ATP assay Kit were used to detect mitochondrial membrane potential (MMP) and energy metabolism, respectively.

Results

In this study, we have successfully established excitotoxic model by KAin vivoandin vitro. KA induced decreased SOD activity and increased MDA concentration. KA cause the change of LC3-II/LC3-I, SQSTM1/p62, and TIGAR expression, indicating the autophagy activation. NADPH plays a protective rolein vivoandin vitro. It reversed the KA-mediated changes in LC3, SQSTM1/p62, TIGAR, and NOX4 protein expression. Mito-apocynin inhibited KA-induced increases in mitochondrial NOX4 expression and activity. Compared with NADPH, the combination showed more significant neuroprotective effects, presenting more neurons survive and better motor function recovery. The combination also better inhibited the over-activated autophagy.In vitro, combination of NADPH and Mito-apocynin performed better in restoring mitochondria membrane potential.

Conclusion

In summary, combined administration of NADPH and NOX inhibitors offers better neuroprotection by reducing NADPH as a NOX substrate to generate ROS. The combined use of NADPH and Mito-apocynin can better restore neurons and mitochondrial function through autophagy pathway.
Benefits of Zebrafish Xenograft Models in Cancer Research
Authors: Chen Xingyu , Li Yongyun , Yao Tengteng , Jia Renbing
Published: February 2021

As a promisingin vivotool for cancer research, zebrafish have been widely applied in various tumor studies. The zebrafish xenograft model is a low-cost, high-throughput tool for cancer research that can be established quickly and requires only a small sample size, which makes it favorite among researchers. Zebrafish patient-derived xenograft (zPDX) models provide promising evidence for short-term clinical treatment. In this review, we discuss the characteristics and advantages of zebrafish, such as their transparent and translucent features, the use of vascular fluorescence imaging, the establishment of metastatic and intracranial orthotopic models, individual pharmacokinetics measurements, and tumor microenvironment. Furthermore, we introduce how these characteristics and advantages are applied other in tumor studies. Finally, we discuss the future direction of the use of zebrafish in tumor studies and provide new ideas for the application of it.
TGF-β-Induced Endothelial to Mesenchymal Transition Is Determined by a Balance Between SNAIL and ID Factors
Authors: Ma Jin , van der Zon Gerard , Gonçalves Manuel A. F. V. , van Dinther Maarten , Thorikay Midory , Sanchez-Duffhues Gonzalo , ten Dijke Peter
Published: February 2021

Endothelial-to-mesenchymal transition (EndMT) plays an important role in embryonic development and disease progression. Yet, how different members of the transforming growth factor-β (TGF-β) family regulate EndMT is not well understood. In the current study, we report that TGF-β2, but not bone morphogenetic protein (BMP)9, triggers EndMT in murine endothelial MS-1 and 2H11 cells. TGF-β2 strongly upregulates the transcription factor SNAIL, and the depletion of Snail is sufficient to abrogate TGF-β2-triggered mesenchymal-like cell morphology acquisition and EndMT-related molecular changes. Although SLUG is not regulated by TGF-β2, knocking out Slug also partly inhibits TGF-β2-induced EndMT in 2H11 cells. Interestingly, in addition to SNAIL and SLUG, BMP9 stimulates inhibitor of DNA binding (ID) proteins. The suppression of Id1, Id2, or Id3 expression facilitated BMP9 in inducing EndMT and, in contrast, ectopic expression of ID1, ID2, or ID3 abrogated TGF-β2-mediated EndMT. Altogether, our results show that SNAIL is critical and indispensable for TGF-β2-mediated EndMT. Although SLUG is also involved in the EndMT process, it plays less of a crucial role in it. In contrast, ID proteins are essential for maintaining endothelial traits and repressing the function of SNAIL and SLUG during the EndMT process. These data suggest that the control over endothelial vs. mesenchymal cell states is determined, at least in part, by a balance between the expression of SNAIL/SLUG and ID proteins.
Plasmalogen-Based Liquid Crystalline Multiphase Structures Involving Docosapentaenoyl Derivatives Inspired by Biological Cubic Membranes
Authors: Angelova Angelina , Angelov Borislav , Drechsler Markus , Bizien Thomas , Gorshkova Yulia E. , Deng Yuru
Published: February 2021

Structural properties of plasmenyl-glycerophospholipids (plasmalogens) have been scarcely studied for plasmalogens with long polyunsaturated fatty acid (PUFA) chains, despite of their significance for the organization and functions of the cellular membranes. Elaboration of supramolecular assemblies involving PUFA-chain plasmalogens in nanostructured mixtures with lyotropic lipids may accelerate the development of nanomedicines for certain severe pathologies (e.g., peroxisomal disorders, cardiometabolic impairments, and neurodegenerative Alzheimer’s and Parkinson’s diseases). Here, we investigate the spontaneous self-assembly of bioinspired, custom-produced docosapentaenoyl (DPA) plasmenyl (ether) and ester phospholipids in aqueous environment (pH 7) by synchrotron small-angle X-ray scattering (SAXS) and cryogenic transmission electron microscopy (cryo-TEM). A coexistence of a liquid crystalline primitive cubic Im3m phase and an inverted hexagonal (HII) phase is observed for the DPA-ethanolamine plasmalogen (C16:1p-22:5n6 PE) derivative. A double-diamond cubic Pn3m phase is formed in mixed assemblies of the phosphoethanolamine plasmalogen (C16:1p-22:5n6 PE) and monoolein (MO), whereas a coexistence of cubic and lamellar liquid crystalline phases is established for the DPA-plasmenyl phosphocholine (C16:1p-22:5n6 PC)/MO mixture at ambient temperature. The DPA-diacyl phosphoinositol (22:5n6-22:5n6 PI) ester lipid displays a propensity for a lamellar phase formation. Double membrane vesicles and multilamellar onion topologies with inhomogeneous distribution of interfacial curvature are formed upon incorporation of the phosphoethanolamine plasmalogen (C16:1p-22:5n6 PE) into dioleoylphosphocholine (DOPC) bilayers. Nanoparticulate formulations of plasmalogen-loaded cubosomes, hexosomes, and various multiphase cubosome- and hexosome-derived architectures and mixed type nano-objects (e.g., oil droplet-embedding vesicles or core–shell particles with soft corona) are produced with PUFA-chain phospholipids and lipophilic antioxidant-containing membrane compositions that are characterized by synchrotron SAXS and cryo-TEM imaging. The obtained multiphase nanostructures reflect the changes in the membrane curvature induced by the inclusion of DPA-based PE and PC plasmalogens, as well as DPA-PI ester derivative, and open new opportunities for exploration of these bioinspired nanoassemblies.
Targeting POLE2 Creates a Novel Vulnerability in Renal Cell Carcinoma via Modulating Stanniocalcin 1
Authors: Zhang Chuanjie , Shen Yan , Gao Lili , Wang Xiaojing , Huang Da , Xie Xin , Xu Danfeng , He Hongchao
Published: February 2021

Objective

The aim of this study is to investigate the biological functions and the underlying mechanisms of DNA polymerase epsilon subunit 2 (POLE2) in renal cell carcinoma (RCC).

Methods

The datasets of POLE2 expression in The Cancer Genome Atlas Kidney Clear Cell Carcinoma (TCGA-KIRC) and International Cancer Genome Consortium (ICGC) databases was selected and the correlation between POLE2 and various clinicopathological parameters was analyzed. The POLE2 expression in RCC tissues was examined by immunohistochemistry. The POLE2 knockdown cell lines were constructed. In vitro and in vivo experiments were carried out to investigate the function of POLE2 on cellular biology of RCC, including cell viability assay, clone formation assay, flow cytometry, wound-healing assay, Transwell assay, qRT-PCR, Western blot, etc. Besides, microarray, co-immunoprecipitation, rescue experiment, and Western blot were used to investigate the molecular mechanisms underlying the functions of POLE2.

Results

POLE2 was overexpressed in RCC tissues, and high expression of POLE2 was correlated with poor prognosis of RCC. Furthermore, knockdown of POLE2 significantly inhibited cell proliferation, migration, and facilitated apoptosis in vitro. In vivo experiments revealed that POLE2 attenuated RCC tumorigenesis and tumor growth. we also illuminated that stanniocalcin 1 (STC1) was a downstream gene of POLE2, which promoted the occurrence and development of RCC. Besides, knockdown of POLE2 significantly upregulated the expression levels of Bad and p21 while the expression levels of HSP70, IGF-I, IGF-II, survivin, and sTNF-R1 were significantly downregulated. Western blot analysis also showed that knockdown of POLE2 inhibited the expression levels of Cancer-related pathway proteins including p-Akt, CCND1, MAPK9, and PIK3CA.

Conclusion

Knockdown of POLE2 attenuates RCC cells proliferation and migration by regulating STC1, suggesting that POLE2-STC1 may become a potential target for RCC therapy.
Beneficial and Detrimental Effects of Reactive Oxygen Species on Lifespan: A Comprehensive Review of Comparative and Experimental Studies
Authors: Shields Hazel J. , Traa Annika , Van Raamsdonk Jeremy M.
Published: February 2021

Aging is the greatest risk factor for a multitude of diseases including cardiovascular disease, neurodegeneration and cancer. Despite decades of research dedicated to understanding aging, the mechanisms underlying the aging process remain incompletely understood. The widely-accepted free radical theory of aging (FRTA) proposes that the accumulation of oxidative damage caused by reactive oxygen species (ROS) is one of the primary causes of aging. To define the relationship between ROS and aging, there have been two main approaches: comparative studies that measure outcomes related to ROS across species with different lifespans, and experimental studies that modulate ROS levels within a single species using either a genetic or pharmacologic approach. Comparative studies have shown that levels of ROS and oxidative damage are inversely correlated with lifespan. While these studies in general support the FRTA, this type of experiment can only demonstrate correlation, not causation. Experimental studies involving the manipulation of ROS levels in model organisms have generally shown that interventions that increase ROS tend to decrease lifespan, while interventions that decrease ROS tend to increase lifespan. However, there are also multiple examples in which the opposite is observed: increasing ROS levels results in extended longevity, and decreasing ROS levels results in shortened lifespan. While these studies contradict the predictions of the FRTA, these experiments have been performed in a very limited number of species, all of which have a relatively short lifespan. Overall, the data suggest that the relationship between ROS and lifespan is complex, and that ROS can have both beneficial or detrimental effects on longevity depending on the species and conditions. Accordingly, the relationship between ROS and aging is difficult to generalize across the tree of life.
Loss of Active Neurogenesis in the Adult Shark Retina
Authors: Hernández-Núñez Ismael , Robledo Diego , Mayeur Hélène , Mazan Sylvie , Sánchez Laura , Adrio Fátima , Barreiro-Iglesias Antón , Candal Eva
Published: February 2021

Neurogenesis is the process by which progenitor cells generate new neurons. As development progresses neurogenesis becomes restricted to discrete neurogenic niches, where it persists during postnatal life. The retina of teleost fishes is thought to proliferate and produce new cells throughout life. Whether this capacity may be an ancestral characteristic of gnathostome vertebrates is completely unknown. Cartilaginous fishes occupy a key phylogenetic position to infer ancestral states fixed prior to the gnathostome radiation. Previous work from our group revealed that the juvenile retina of the catshark Scyliorhinus canicula, a cartilaginous fish, shows active proliferation and neurogenesis. Here, we compared the morphology and proliferative status of the retina in catshark juveniles and adults. Histological and immunohistochemical analyses revealed an important reduction in the size of the peripheral retina (where progenitor cells are mainly located), a decrease in the thickness of the inner nuclear layer (INL), an increase in the thickness of the inner plexiform layer and a decrease in the cell density in the INL and in the ganglion cell layer in adults. Contrary to what has been reported in teleost fish, mitotic activity in the catshark retina was virtually absent after sexual maturation. Based on these results, we carried out RNA-Sequencing (RNA-Seq) analyses comparing the retinal transcriptome of juveniles and adults, which revealed a statistically significant decrease in the expression of many genes involved in cell proliferation and neurogenesis in adult catsharks. Our RNA-Seq data provides an excellent resource to identify new signaling pathways controlling neurogenesis in the vertebrate retina.
Restoration of FVIII Function and Phenotypic Rescue in Hemophilia A Mice by Transplantation of MSCs Derived From F8-Modified iPSCs
Authors: Qiu Liyan , Xie Mi , Zhou Miaojin , Liu Xionghao , Hu Zhiqing , Wu Lingqian
Published: February 2021

Hemophilia A (HA), an X-linked recessive congenital bleeding disorder, affects 80%–85% of patients with hemophilia. Nearly half of severe cases of hemophilia are caused by a 0.6-Mb genomic inversion (Inv22) that disrupts F8. Although viral-based gene therapy has shown therapeutic effects for hemophilia B (HB), this promising approach is not applicable for HA at the present stage; this limitation is mainly due to the large size of F8 cDNA, which far exceeds the adeno-associated virus (AAV) packaging capacity. We previously reported an in situ genetic correction of Inv22 in HA patient-specific induced pluripotent stem cells (HA-iPSCs) by using TALENs. We also investigated an alternative strategy for targeted gene addition, in which cDNA of the B-domain deleted F8 (BDDF8) was targeted at the rDNA locus of HA-iPSCs using TALENickases to restore FVIII function. Mesenchymal stem cells (MSCs) have low immunogenicity and can secrete FVIII under physiological conditions; in this study, MSCs were differentiated from F8-corrected iPSCs, BDDF8-iPSCs, and HA-iPSCs. Differentiated MSCs were characterized, and FVIII expression efficacy in MSCs was verified in vitro. The three types of MSCs were introduced into HA mice via intravenous injection. Long-term engraftment with restoration of FVIII function and phenotypic rescue was observed in HA mice transplanted with F8-corrected iMSCs and BDDF8-iMSCs. Our findings suggest that ex vivo gene therapy using iMSCs derived from F8-modified iPSCs can be feasible, effective, and promising for the clinical translation of therapeutic gene editing of HA and other genetic birth defects, particularly those that involve large sequence variants.
Impaired Barrier Function and Immunity in the Colon of Aldo-Keto Reductase 1B8 Deficient Mice
Authors: Wang Xin , Khoshaba Ramina , Shen Yi , Cao Yu , Lin Minglin , Zhu Yun , Cao Zhe , Liao Duan-Fang , Cao Deliang
Published: February 2021

Aldo-keto reductase 1B10 (AKR1B10) is downregulated in human ulcerative colitis (UC) and colorectal cancer, being a potential pathogenic factor of these diseases. Aldo-keto reductase 1B8 (AKR1B8) is the ortholog in mice of human AKR1B10. Targeted AKR1B8 deficiency disrupts homeostasis of epithelial self-renewal and leads to susceptibility to colitis and carcinogenesis. In this study, we found that in AKR1B8 deficient mice, Muc2 expression in colon was diminished, and permeability of colonic epithelium increased. Within 24 h, orally administered FITC-dextran penetrated into mesenteric lymph nodes (MLN) and liver in AKR1B8 deficient mice, but not in wild type controls. In the colon of AKR1B8 deficient mice, neutrophils and mast cells were markedly infiltrated, γδT cells were numerically and functionally impaired, and dendritic cell development was altered. Furthermore, Th1, Th2, and Th17 cells decreased, but Treg and CD8T cells increased in the colon and MLN of AKR1B8 deficient mice. In colonic epithelial cells of AKR1B8 deficient mice, p-AKT (T308 and S473), p-ERK1/2, p-IKBα, p-p65 (S536), and IKKα expression decreased, accompanied with downregulation of IL18 and CCL20 and upregulation of IL1β and CCL8. These data suggest AKR1B8 deficiency leads to abnormalities of intestinal epithelial barrier and immunity in colon.
RAMP2-AS1 Regulates Endothelial Homeostasis and Aging
Authors: Lai Chih-Hung , Chen Aleysha T. , Burns Andrew B. , Sriram Kiran , Luo Yingjun , Tang Xiaofang , Branciamore Sergio , O’Meally Denis , Chang Szu-Ling , Huang Po-Hsun , Shyy John Y-J. , Chien Shu , Rockne Russell C. , Chen Zhen Bouman
Published: February 2021

The homeostasis of vascular endothelium is crucial for cardiovascular health and endothelial cell (EC) aging and dysfunction could negatively impact vascular function. Leveraging transcriptome profiles from ECs subjected to various stimuli, including time-series data obtained from ECs under physiological pulsatile flow vs. pathophysiological oscillatory flow, we performed principal component analysis (PCA) to identify key genes contributing to divergent transcriptional states of ECs. Through bioinformatics analysis, we identified that a long non-coding RNA (lncRNA) RAMP2-AS1 encoded on the antisense of RAMP2, a determinant of endothelial homeostasis and vascular integrity, is a novel regulator essential for EC homeostasis and function. Knockdown of RAMP2-AS1 suppressed RAMP2 expression and caused EC functional changes promoting aging, including impaired angiogenesis and increased senescence. Our study demonstrates an integrative approach to quantifying EC aging based on transcriptome changes, which also identified a number of novel regulators, including protein-coding genes and many lncRNAs involved EC functional modulation, exemplified by RAMP2-AS1.
Mechanistic and Therapeutic Implications of Extracellular Vesicles as a Potential Link Between Covid-19 and Cardiovascular Disease Manifestations
Authors: Pironti Gianluigi , Andersson Daniel C. , Lund Lars H.
Published: February 2021

Extracellular vesicles (EVs), which are cell released double layered membrane particles, have been found in every circulating body fluid, and provide a tool for conveying diverse information between cells, influencing both physiological and pathological conditions. Viruses can hijack the EVs secretory pathway to exit infected cells and use EVs endocytic routes to enter uninfected cells, suggesting that EVs and viruses can share common cell entry and biogenesis mechanisms. SARS-CoV-2 is responsible of the coronavirus disease 2019 (Covid-19), which may be accompanied by severe multi-organ manifestations. EVs may contribute to virus spreading via transfer of virus docking receptors such as CD9 and ACE2. Covid-19 is known to affect the renin angiotensin system (RAS), and could promote secretion of harmful EVs. In this scenario EVs might be linked to cardiovascular manifestations of the Covid-19 disease through unbalance in RAS. In contrast EVs derived from mesenchymal stem cells or cardiosphere derived cells, may promote cardiovascular function due to their beneficial effect on angiogenesis, fibrosis, contractility and immuno-modulation. In this article we assessed the potential impact of EVs in cardiovascular manifestations of Covid-19 and highlight potential strategies to control the extracellular signaling for future therapies.