bluuebunny/crossref_metadata_2025 huggingface dataset:
Lactic Acid Bacteria Diversity and Characterization of Probiotic Candidates in Fermented Meats
Authors: Parlindungan Elvina , Lugli Gabriele A. , Ventura Marco , van Sinderen Douwe , Mahony Jennifer
Published: July 2021

Probiotics are defined as live microorganisms which confer health benefits to the host when administered in adequate amounts. Many lactic acid bacteria (LAB) strains have been classified as probiotics and fermented foods are an excellent source of such LAB. In this study, novel probiotic candidates from two fermented meats (pancetta and prosciutto) were isolated and characterized. LAB populations present in pancetta and prosciutto were evaluated and Lactiplantibacillus plantarum was found to be the dominant species. The antagonistic ability of selected isolates against LAB and non-LAB strains was investigated, in particular, the ability to produce anti-microbial compounds including organic acids and bacteriocins. Probiotic characteristics including antibiotic susceptibility, hydrophobicity and autoaggregation capacity; and ability to withstand simulated gastric juice, bile salt, phenol and NaCl were assessed. Among the characterized strains, L. plantarum 41G isolated from prosciutto was identified as the most robust probiotic candidate compared. Results from this study demonstrate that artisanal fermented meat is a rich source of novel strains with probiotic potential.
An Inverse Source Problem for the Generalized Subdiffusion Equation with Nonclassical Boundary Conditions
Authors: Bazhlekova Emilia
Published: July 2021

An initial-boundary-value problem is considered for the one-dimensional diffusion equation with a general convolutional derivative in time and nonclassical boundary conditions. We are concerned with the inverse source problem of recovery of a space-dependent source term from given final time data. Generalized eigenfunction expansions are used with respect to a biorthogonal pair of bases. Existence, uniqueness and stability estimates in Sobolev spaces are established.
Aronofsky’s Black Swan as a Postmodern Fairy Tale: Mirroring a Narcissistic Society
Authors: Landwehr Margarete Johanna
Published: July 2021

Based on the plot of Swan Lake, Black Swan depicts an ingenue’s metamorphosis into a woman and a prima ballerina that contains a fairy-tale plot in which a naïve heroine overcomes enemies and obstacles in order to achieve success and sexual maturity. Unlike a traditional fairy tale, this cinematic tale concludes with death and the clear distinctions between good and evil, helper and adversary and reality vs. fantasy are fluid. As in many fairy tales, the film criticizes the values of its era, namely, the narcissistic aspects of contemporary society with its excessive worship of youth, beauty and celebrity, and its most pernicious results—escape into fantasy and insanity, aggressive rivalry, violence, and self-destruction.
Associations between Hospital Quality Outcomes and Medicare Spending per Beneficiary in the USA
Authors: Cook Karyn , Foster Brent , Perry I’sis , Hoke Christy , Smith Dana , Peterson Leanne , Martin John , Korvink Michael , Gunn Laura H.
Published: July 2021

The cost of healthcare in the United States has increased over time. However, patient health outcomes have not trended with spending. There is a need to better comprehend the association between healthcare costs in the United States and hospital quality outcomes. Medicare spending per beneficiary (MSPB), a homogeneous metric across providers, can be used to evaluate the association between episodic Medicare spending and quality of care. Fifteen inpatient outcome measures were selected from Hospital Compare data among all (n = 4758) facilities and transformed to quintiles to ensure comparability across measures and to reduce the influence of outliers on the analysis. Both univariate and multiresponse multinomial ordered probit regression models were utilized across outcome domains to quantify associations between outcomes and spending. We found that MSPB was not associated with quality of care in most cases, adding evidence of a lack of outcome accountability among Medicare-funded facilities. Furthermore, worse outcomes were found to be associated with increased spending for some metrics. Policies are needed to align quality of care outcomes with the increasing costs of U.S. healthcare.
Non-Coding RNAs in the Cardiac Action Potential and Their Impact on Arrhythmogenic Cardiac Diseases
Authors: Lozano-Velasco Estefania , Aranega Amelia , Franco Diego
Published: July 2021

Cardiac arrhythmias are prevalent among humans across all age ranges, affecting millions of people worldwide. While cardiac arrhythmias vary widely in their clinical presentation, they possess shared complex electrophysiologic properties at cellular level that have not been fully studied. Over the last decade, our current understanding of the functional roles of non-coding RNAs have progressively increased. microRNAs represent the most studied type of small ncRNAs and it has been demonstrated that miRNAs play essential roles in multiple biological contexts, including normal development and diseases. In this review, we provide a comprehensive analysis of the functional contribution of non-coding RNAs, primarily microRNAs, to the normal configuration of the cardiac action potential, as well as their association to distinct types of arrhythmogenic cardiac diseases.
The Approach to Thrombosis Prevention across the Spectrum of Philadelphia-Negative Classic Myeloproliferative Neoplasms
Authors: Koschmieder Steffen
Published: July 2021

Patients with myeloproliferative neoplasm (MPN) are potentially facing diminished life expectancy and decreased quality of life, due to thromboembolic and hemorrhagic complications, progression to myelofibrosis or acute leukemia with ensuing signs of hematopoietic insufficiency, and disturbing symptoms such as pruritus, night sweats, and bone pain. In patients with essential thrombocythemia (ET) or polycythemia vera (PV), current guidelines recommend both primary and secondary measures to prevent thrombosis. These include acetylsalicylic acid (ASA) for patients with intermediate- or high-risk ET and all patients with PV, unless they have contraindications for ASA use, and phlebotomy for all PV patients. A target hematocrit level below 45% is demonstrated to be associated with decreased cardiovascular events in PV. In addition, cytoreductive therapy is shown to reduce the rate of thrombotic complications in high-risk ET and high-risk PV patients. In patients with prefibrotic primary myelofibrosis (pre-PMF), similar measures are recommended as in those with ET. Patients with overt PMF may be at increased risk of bleeding and thus require a more individualized approach to thrombosis prevention. This review summarizes the thrombotic risk factors and primary and secondary preventive measures against thrombosis in MPN.
Comparative Transcriptomic Analysis of Differentially Expressed Transcripts Associated with Flowering Time of Loquat (Eriobotya japonica Lindl.)
Authors: An Haishan , Jiang Shuang , Zhang Jiaying , Xu Fangjie , Zhang Xueying
Published: July 2021

Flowering is an important phenophase of plant species, however, knowledge about the regulatory mechanism controlling flowering cues in loquat is limited. To identify candidate genes regulating flowering time in loquat, we used RNA-Seq technology to conduct a comparative transcriptome analysis of differentiating apical buds collected from the early-flowering variety ‘Baiyu’ and the late-flowering variety ‘Huoju’. A total of 28,842 differentially expressed transcripts (DETs) were identified. Of these, 42 DETs controlled flowering time while 17 other DETs were associated with the ABA signaling pathway. Compared with those in ‘Huoju’, EjFT, EjFY, EjFLK, and EjCAL1-like were significantly upregulated in ‘Baiyu’. Moreover, transcripts of the ABA 8′-hydroxylases (EjABH2, EjABH4, and EjABH4-like2), the ABA receptors (EjPYL4/8), and the bZIP transcription factor EjABI5-like were upregulated in ‘Baiyu’ compared with ‘Huoju’. Hence, they might regulate loquat flowering time. There was no significant difference between ‘Baiyu’ and ‘Huoju’ in terms of IAA content. However, the ABA content was about ten-fold higher in the apical buds of ‘Baiyu’ than in those of ‘Huoju’. The ABA:IAA ratio sharply rose and attained a peak during bud differentiation. Thus, ABA is vital in regulating floral bud formation in loquat. The results of the present study help clarify gene transcription during loquat flowering.
Parental Warmth, Gratitude, and Prosocial Behavior among Chinese Adolescents: The Moderating Effect of School Climate
Authors: Luo Haocheng , Liu Qingqi , Yu Chengfu , Nie Yangang
Published: July 2021

Parental warmth plays an important role in the development of adolescents’ physical and mental health. There are numerous empirical studies indicating a relationship between parental warmth and prosocial behavior among adolescents, although the underlying mechanisms of this association remain unclear. Adopting a longitudinal design across two time points, the present study proposes a moderated mediation model to explore the mediating role of gratitude and the moderating role of the school climate between parental warmth and prosocial behavior. The sample consisted of 934 participants (483 boys and 451 girls) who participated in the second assessment and completed questionnaires assessing gratitude, school climate, and prosocial behavior in April 2019. Their parents participated in the first assessment and completed a questionnaire pertaining to parental warmth in October 2018. After controlling for the gender and age of the adolescents, the results showed that the positive association between parental warmth and prosocial behavior is mediated by gratitude, and school climate does play a moderating role in the second half of the mediating path. Specifically, the school climate can play a protective role in adolescents with low levels of gratitude. For adolescents with less gratitude, a strong school climate can promote more prosocial behaviors and can effectively alleviate the negative prediction of low levels of gratitude. This study provides a theoretical explanation for the generation of adolescents’ prosocial behavior and provides theoretical guidance for the interventions of schools and parents.
Exploration of Mental Readiness for Enhancing Dentistry in an Inter-Professional Climate
Authors: McDonald Judy M. , Paganelli Corrado
Published: July 2021

Competencies required for dentistry go far beyond the academic or scientific spheres. They incorporate important mental readiness concepts at its core with an appropriate balance of operational readiness (i.e., technical, physical, mental readiness). The aim of this exploratory study was to investigate the importance of mental readiness for optimal performance in the daily challenges faced by dentists using an Operational Readiness Framework. One-on-one interviews were conducted with a select group of seasoned dentists to determine their mental readiness before, during and after successfully performing in challenging situations. Quantitative and qualitative analyses of mental readiness were applied. Study findings were compared with a Wheel of Excellence based on results from other high-performance domains such as surgery, policing, social services and Olympic athletics. The analysis revealed that specific mental practices are required to achieve peak performance, and the balance between physical, technical and mental readiness underpins these dentists’ competency. Common elements of success were found—commitment, confidence, visualization, mental preparation, focus, distraction control, and evaluation and coping. This exploration confirmed many similarities in mental readiness practices engaged across high-risk professions. Universities, clinics and hospitals are looking for innovative ways to build teamwork and capacity through inter-professional collaboration. Results from these case studies warrant further investigation and may be significant enough to stimulate innovative curriculum design. Based on these preliminary dentistry findings, three training/evaluation tools from other professions in population health were adapted to demonstrate future application.
Prevalence and Associated Factors of Falls among Older Adults between Urban and Rural Areas of Shantou City, China
Authors: Chen Xiaodong , Lin Zeting , Gao Ran , Yang Yijian , Li Liping
Published: July 2021

Background: To investigate the prevalence of falls and associated factors among older adults in urban and rural areas and to facilitate the design of fall prevention interventions. Methods: We used cluster random sampling to investigate the sociodemographic information, living habits, medical status, falls, home environment, and balance ability among 649 older adult participants. Univariate and multivariate logistic regression were used to examine the associated factors of falls. Results: The incidence of falls among older adults in Shantou City was 20.65%. Among them, the incidence was 27.27% in urban areas and 16.99% in rural areas. The rate of injury from falls among older adults was 14.48%, with18.61% in urban area and 12.20% in rural area. Multivariate analysis showed that the associated factors of falls among older adults in Shantou City included a high school or below education level (OR = 2.387, 95% CI: 1.305–4.366); non-farming as the previous occupation (OR = 2.574, 95% CI: 1.613–4.109); incontinence(OR = 2.881, 95% CI: 1.517–5.470); lack of fall prevention education (OR = 1.856, 95% CI: 1.041–3.311); and reduced balance ability (OR = 3.917, 95% CI: 2.532–6.058). Discussion: Older adults have a higher rate of falling in Shantou City, compared to the average rate in China. There are similarities and differences in the associated factors of falls among older adults between urban and rural areas of Shantou City. Targeted interventions for older adults in different regions may be more effective in reducing the risk of falls.
Regulation of Mindfulness-Based Music Listening on Negative Emotions Related to COVID-19: An ERP Study
Authors: Liu Xiaolin , Liu Yong , Shi Huijuan , Li Ling , Zheng Maoping
Published: July 2021

The current study aimed to explore the behavioral and neural correlates of mindfulness-based music listening regulation of induced negative emotions related to COVID-19 using the face–word Stroop task. Eighty-five young adults visited the laboratory and were randomly assigned to three groups: a calm music group (CMG: n = 28), a happy music group (HMG: n = 30), and a sad music group (SMG: n = 27). Negative emotions were induced in all participants using a COVID-19 video, followed by the music intervention condition. Participants underwent the face–word Stroop tasks during which event-related potentials (ERPs) were recorded. The N2, N3, P3, and late positive component (LPC) were investigated. The results showed that calm music and happy music effectively regulate young adults’ induced negative emotions, while young adults experienced more negative emotions when listening to sad music; the negative mood states at the post-induction phase inhibited the reaction of conflict control in face–word Stroop tasks, which manifested as lower accuracy (ACC) and slower reaction times (RTs). ERP results showed negative mood states elicited greater N2, N3, and LPC amplitudes and smaller P3 amplitudes. Further studies are needed to develop intervention strategies to enhance emotion regulation related to COVID-19 for other groups.
Intellectual Structure and Evolution of Accounting Conservatism Research: Past Trends and Future Research Suggestions
Authors: Bhutta Umair , Martins Jéssica Nunes , Mata Mário Nuno , Raza Ali , Dantas Rui Miguel , Correia Anabela Batista , Rafiq Muhammad
Published: July 2021

Accounting conservatism (AC) is one of the components of financial reporting, and has been widely studied by academicians to identify its impact on information quality. Scholars in accounting have started to explore how AC is related to different fundamental functional areas of organizations. The interest of the scholars has resulted in an increasing number of publications in this field. In this study, we examined 408 indexed publications related to AC. This work’s objectives include analyzing the regional distribution, size, and evolution of this knowledge base by identifying key authors, documents, and journals while exploring current literature, scholarly structure, and highlighting contemporary trends. The findings of the study concluded that most of the studies are conducted in developed nations contexts. However, there are still areas that need further exploration to obtain more profound insights on the subject. This bibliometric review inspires a new generation of researchers on the topic by giving them an overview of the past studies related to AC.
A Trajectory Privacy Protection Method Based on Random Sampling Differential Privacy
Authors: Ma Tinghuai , Song Fagen
Published: July 2021

With the popularity of location-aware devices (e.g., smart phones), a large number of trajectory data were collected. The trajectory dataset can be used in many fields including traffic monitoring, market analysis, city management, etc. The collection and release of trajectory data will raise serious privacy concerns for users. If users’ privacy is not protected enough, they will refuse to share their trajectory data. In this paper, a new trajectory privacy protection method based on random sampling differential privacy (TPRSDP), which can provide more security protection, is proposed. Compared with other methods, it takes less time to run this method. Experiments are conducted on two real world datasets to validate the proposed scheme, and the results are compared with others in terms of running time and information loss. The performance of the scheme with different parameter values is verified. The setting of the new scheme parameters is discussed in detail, and some valuable suggestions are given.
Synergistic Effect of the Long-Term Overexpression of Bcl-2 and BDNF Lentiviral in Cell Protecting against Death and Generating TH Positive and CHAT Positive Cells from MSC
Authors: Borkowska Paulina , Zielinska Aleksandra , Paul-Samojedny Monika , Stojko Rafał , Kowalski Jan
Published: July 2021

Mesenchymal stem cells (MSC) are potentially a good material for transplantation in many diseases, including neurodegenerative diseases. The main problem with using them is the low percentage of surviving cells after the transplant procedure and the naturally poor ability of MSC to spontaneously differentiate into certain types of cells, which results in their poor integration with the host cells. The aim and the novelty of this work consists in the synergistic overexpression of two genes, BCL2 and BDNF, using lentiviral vectors. According to our hypothesis, the overexpression of the BCL2 gene is aimed at increasing the resistance of cells to stressors and toxic factors. In turn, the overexpression of the BDNF gene is suspected to direct the MSC into the neural differentiation pathway. As a result, it was shown that the overexpression of both genes and the overproduction of proteins is permanent and persists for at least 60 days. The synergistically transduced MSC were significantly more resistant to the action of staurosporine; 12 days after transduction, the synergistically transduced MSC had a six-times greater survival rate. The overexpression of the Bcl-2 and BDNF proteins was sufficient to stimulate a significant overexpression of the CHAT gene, and under specific conditions, the TH, TPH1, and SYP genes were also overexpressed. Modified MSC are able to differentiate into cholinergic and dopaminergic neurons, and the release of acetylcholine and dopamine may indicate their functionality.
In Vitro Osteoinductivity Assay of Hydroxylapatite Scaffolds, Obtained with Biomorphic Transformation Processes, Assessed Using Human Adipose Stem Cell Cultures
Authors: Iaquinta Maria Rosa , Torreggiani Elena , Mazziotta Chiara , Ruffini Andrea , Sprio Simone , Tampieri Anna , Tognon Mauro , Martini Fernanda , Mazzoni Elisa
Published: July 2021

In this study, the in vitro biocompatibility and osteoinductive ability of a recently developed biomorphic hydroxylapatite ceramic scaffold (B-HA) derived from transformation of wood structures were analyzed using human adipose stem cells (hASCs). Cell viability and metabolic activity were evaluated in hASCs, parental cells and in recombinant genetically engineered hASC-eGFP cells expressing the green fluorescence protein. B-HA osteoinductivity properties, such as differentially expressed genes (DEG) involved in the skeletal development pathway, osteocalcin (OCN) protein expression and mineral matrix deposition in hASCs, were evaluated. In vitro induction of osteoblastic genes, such as Alkaline phosphatase (ALPL), Bone gamma-carboxyglutamate (gla) protein (BGLAP), SMAD family member 3 (SMAD3), Sp7 transcription factor (SP7) and Transforming growth factor, beta 3 (TGFB3) and Tumor necrosis factor (ligand) superfamily, member 11 (TNFSF11)/Receptor activator of NF-κB (RANK) ligand (RANKL), involved in osteoclast differentiation, was undertaken in cells grown on B-HA. Chondrogenic transcription factor SRY (sex determining region Y)-box 9 (SOX9), tested up-regulated in hASCs grown on the B-HA scaffold. Gene expression enhancement in the skeletal development pathway was detected in hASCs using B-HA compared to sintered hydroxylapatite (S-HA). OCN protein expression and calcium deposition were increased in hASCs grown on B-HA in comparison with the control. This study demonstrates the biocompatibility of the novel biomorphic B-HA scaffold and its potential use in osteogenic differentiation for hASCs. Our data highlight the relevance of B-HA for bone regeneration purposes.
Targeting Leukemic Stem Cells in Chronic Myeloid Leukemia: Is It Worth the Effort?
Authors: Soverini Simona , De Santis Sara , Monaldi Cecilia , Bruno Samantha , Mancini Manuela
Published: July 2021

Chronic myeloid leukemia (CML) is a classical example of stem cell cancer since it arises in a multipotent hematopoietic stem cell upon the acquisition of the t(9;22) chromosomal translocation, that converts it into a leukemic stem cell (LSC). The resulting BCR-ABL1 fusion gene encodes a deregulated tyrosine kinase that is recognized as the disease driver. Therapy with tyrosine kinase inhibitors (TKIs) eliminates progenitor and more differentiated cells but fails to eradicate quiescent LSCs. Thus, although many patients obtain excellent responses and a proportion of them can even attempt treatment discontinuation (treatment free remission [TFR]) after some years of therapy, LSCs persist, and represent a potentially dangerous reservoir feeding relapse and hampering TFR. Over the past two decades, intensive efforts have been devoted to the characterization of CML LSCs and to the dissection of the cell-intrinsic and -extrinsic mechanisms sustaining their persistence, in an attempt to find druggable targets enabling LSC eradication. Here we provide an overview and an update on these mechanisms, focusing in particular on the most recent acquisitions. Moreover, we provide a critical appraisal of the clinical relevance and feasibility of LSC targeting in CML.
New Concept and Apparatus for Cytocentrifugation and Cell Processing for Microscopy Analysis
Authors: Ligasová Anna , Koberna Karel
Published: July 2021

Cytocentrifugation is a common technique for the capture of cells on microscopic slides. It usually requires a special cytocentrifuge or cytorotor and cassettes. In the study presented here, we tested the new concept of cytocentrifugation based on the threaded connection of the lid and the sample holder to ensure an adjustable flow of solutions through the filters and the collection of the filtered solutions in the reservoir during centrifugation. To test this concept, we developed a device for the preparation of cell samples on circular coverslips. The device was tested for the capture and sample processing of both eukaryotic and prokaryotic cells, cell nuclei, and mitochondria for microscopy analysis including image cytometry. Moreover, an efficient procedure was developed for capturing formaldehyde-fixed cells on non-treated coverslips without cell drying. The results showed that the tested arrangement enables the effective capture and processing of all of the tested samples and the developed device represents an inexpensive alternative to common cytocentrifuges, as only the paper filter is consumed during sample processing, and no special centrifuge, cytorotor, or cassette is necessary. As no additional system of solution removal is required during sample staining, the tested concept also facilitates the eventual automation of the staining procedure.
Diethylstilbestrol, a Novel ANO1 Inhibitor, Exerts an Anticancer Effect on Non-Small Cell Lung Cancer via Inhibition of ANO1
Authors: Seo Yohan , Jeong Sung Baek , Woo Joo Han , Kwon Oh-Bin , Lee Sion , Oh Hye In , Jo Sungwoo , Park Seon Ju , Namkung Wan , Moon Uk Yeol , Lee Sungwoo
Published: July 2021

Non-small cell lung cancer (NSCLC) is one of the leading causes of cancer-related mortality; thus, therapeutic targets continue to be developed. Anoctamin1 (ANO1), a novel drug target considered for the treatment of NSCLC, is a Ca2+-activated chloride channel (CaCC) overexpressed in various carcinomas. It plays an important role in the development of cancer; however, the role of ANO1 in NSCLC is unclear. In this study, diethylstilbestrol (DES) was identified as a selective ANO1 inhibitor using high-throughput screening. We found that DES inhibited yellow fluorescent protein (YFP) fluorescence reduction caused by ANO1 activation but did not inhibit cystic fibrosis transmembrane conductance regulator channel activity or P2Y activation-related cytosolic Ca2+ levels. Additionally, electrophysiological analyses showed that DES significantly reduced ANO1 channel activity, but it more potently reduced ANO1 protein levels. DES also inhibited the viability and migration of PC9 cells via the reduction in ANO1, phospho-ERK1/2, and phospho-EGFR levels. Moreover, DES induced apoptosis by increasing caspase-3 activity and PARP-1 cleavage in PC9 cells, but it did not affect the viability of hepatocytes. These results suggest that ANO1 is a crucial target in the treatment of NSCLC, and DES may be developed as a potential anti-NSCLC therapeutic agent.
From Affinity to Proximity Techniques to Investigate Protein Complexes in Plants
Authors: Kerbler Sandra M. , Natale Roberto , Fernie Alisdair R. , Zhang Youjun
Published: July 2021

The study of protein–protein interactions (PPIs) is fundamental in understanding the unique role of proteins within cells and their contribution to complex biological systems. While the toolkit to study PPIs has grown immensely in mammalian and unicellular eukaryote systems over recent years, application of these techniques in plants remains under-utilized. Affinity purification coupled to mass spectrometry (AP-MS) and proximity labeling coupled to mass spectrometry (PL-MS) are two powerful techniques that have significantly enhanced our understanding of PPIs. Relying on the specific binding properties of a protein to an immobilized ligand, AP is a fast, sensitive and targeted approach used to detect interactions between bait (protein of interest) and prey (interacting partners) under near-physiological conditions. Similarly, PL, which utilizes the close proximity of proteins to identify potential interacting partners, has the ability to detect transient or hydrophobic interactions under native conditions. Combined, these techniques have the potential to reveal an unprecedented spatial and temporal protein interaction network that better understands biological processes relevant to many fields of interest. In this review, we summarize the advantages and disadvantages of two increasingly common PPI determination techniques: AP-MS and PL-MS and discuss their important application to plant systems.
Mesenchymal Stem Cell-Derived Small Extracellular Vesicles Protect Cardiomyocytes from Doxorubicin-Induced Cardiomyopathy by Upregulating Survivin Expression via the miR-199a-3p-Akt-Sp1/p53 Signaling Pathway
Authors: Lee Ji Yoon , Chung Jihwa , Byun Yeongju , Kim Kyoung Hwa , An Shung Hyun , Kwon Kihwan
Published: July 2021

Cardiotoxicity is associated with the long-term clinical application of doxorubicin (DOX) in cancer patients. Mesenchymal stem cell-derived small extracellular vesicles (MSC-sEVs) including exosomes have been suggested for the treatment of various diseases, including ischemic diseases. However, the effects and functional mechanism of MSC-sEVs in DOX-induced cardiomyopathy have not been clarified. Here, MSC-sEVs were isolated from murine embryonic mesenchymal progenitor cell (C3H/10T1/2) culture media, using ultrafiltration. H9c2 cardiac myoblast cells were pretreated with MSC-sEVs and then exposed to DOX. For in vivo studies, male C57BL/6 mice were administered MSC-sEVs intravenously, prior to a single dose of DOX (15 mg/kg, intraperitoneal). The mice were sacrificed 14 days after DOX treatment. The results showed that MSC-sEVs protected cardiomyocytes from DOX-induced cell death. H9c2 cells treated with DOX showed downregulation of both phosphorylated Akt and survivin, whereas the treatment of MSC-sEVs recovered expression, indicating their anti-apoptotic effects. Three microRNAs (miRNAs) (miR 199a-3p, miR 424-5p, and miR 21-5p) in MSC-sEVs regulated the Akt-Sp1/p53 signaling pathway in cardiomyocytes. Among them, miR 199a-3p was involved in regulating survivin expression, which correlated with the anti-apoptotic effects of MSC-sEVs. In in vivo studies, the echocardiographic results showed that the group treated with MSC-sEVs recovered from DOX-induced cardiomyopathy, showing improvement of both the left ventricle fraction and ejection fraction. MSC-sEVs treatment also increased both survivin and B-cell lymphoma 2 expression in heart tissue compared to the DOX group. Our results demonstrate that MSC-sEVs have protective effects against DOX-induced cardiomyopathy by upregulating survivin expression, which is mediated by the regulation of Akt activation by miRNAs in MSC-sEVs. Thus, MSC-sEVs may be a novel therapy for the prevention of DOX-induced cardiomyopathy.
Evaluation of Chemical Changes in Laboratory-Induced Colistin-Resistant Klebsiella pneumoniae
Authors: Pruss Agata , Kwiatkowski Paweł , Łopusiewicz Łukasz , Masiuk Helena , Sobolewski Peter , Fijałkowski Karol , Sienkiewicz Monika , Smolak Adam , Giedrys-Kalemba Stefania , Dołęgowska Barbara
Published: July 2021

This study evaluates the electrical potential and chemical alterations in laboratory-induced colistin-resistant Klebsiella pneumoniae, as compared to the susceptible strain using spectroscopic analyses. The minimal inhibitory concentration (MIC) of colistin, ζ-potential and chemical composition analysis of K. pneumoniae strains are determined. The results obtained for the K. pneumoniaeCol-R with induced high-level colistin resistance (MIC = 16.0 ± 0.0 mg/L) are compared with the K. pneumoniaeCol-S strain susceptible to colistin (MIC = 0.25 ± 0.0 mg/L). Fourier transform infrared (FTIR) and Raman spectroscopic studies revealed differences in bacterial cell wall structures and lipopolysaccharide (LPS) of K. pneumoniaeCol-R and K. pneumoniaeCol-S strains. In the beginning, we assumed that the obtained results could relate to a negative charge of the bacterial surface and different electrostatic interactions with cationic antibiotic molecules, reducing the affinity of colistin and leading to its lower penetration into K. pneumoniaeCol-R cell. However, no significant differences in the ζ-potential between the K. pneumoniaeCol-R and K. pneumoniaeCol-S strains are noticed. In conclusion, this mechanism is most probably associated with recognisable changes in the chemical composition of the K. pneumoniaeCol-R cell wall (especially in LPS) when compared to the susceptible strain.
Synthesis of pH and Glucose Responsive Silk Fibroin Hydrogels
Authors: Tao Xiaosheng , Jiang Fujian , Cheng Kang , Qi Zhenzhen , Yadavalli Vamsi K. , Lu Shenzhou
Published: July 2021

Silk fibroin (SF) has attracted much attention due to its high, tunable mechanical strength and excellent biocompatibility. Imparting the ability to respond to external stimuli can further enhance its scope of application. In order to imbue stimuli-responsive behavior in silk fibroin, we propose a new conjugated material, namely cationic SF (CSF) obtained by chemical modification of silk fibroin with ε-Poly-(L-lysine) (ε-PLL). This pH-responsive CSF hydrogel was prepared by enzymatic crosslinking using horseradish peroxidase and H2O2. Zeta potential measurements and SDS-PAGE gel electrophoresis show successful synthesis, with an increase in isoelectric point from 4.1 to 8.6. Fourier transform infrared (FTIR) and X-ray diffraction (XRD) results show that the modification does not affect the crystalline structure of SF. Most importantly, the synthesized CSF hydrogel has an excellent pH response. At 10 wt.% ε-PLL, a significant change in swelling with pH is observed. We further demonstrate that the hydrogel can be glucose-responsive by the addition of glucose oxidase (GOx). At high glucose concentration (400 mg/dL), the swelling of CSF/GOx hydrogel is as high as 345 ± 16%, while swelling in 200 mg/dL, 100 mg/dL and 0 mg/dL glucose solutions is 237 ± 12%, 163 ± 12% and 98 ± 15%, respectively. This shows the responsive swelling of CSF/GOx hydrogels to glucose, thus providing sufficient conditions for rapid drug release. Together with the versatility and biological properties of fibroin, such stimuli-responsive silk hydrogels have great potential in intelligent drug delivery, as soft matter substrates for enzymatic reactions and in other biomedical applications.
Antithrombotic Therapy in Patients with Peripheral Artery Disease: A Focused Review on Oral Anticoagulation
Authors: Rivera-Caravaca José Miguel , Camelo-Castillo Anny , Ramírez-Macías Inmaculada , Gil-Pérez Pablo , López-García Cecilia , Esteve-Pastor María Asunción , Orenes-Piñero Esteban , Tello-Montoliu Antonio , Marín Francisco
Published: July 2021

Peripheral artery disease (PAD) is a major cause of morbidity and mortality but it is usually underdiagnosed and undertreated. Patients with PAD present dysregulated procoagulant, anticoagulant, and fibrinolytic pathways leading to arterial and venous thrombosis. The risk of several ischemic-related complications could be mitigated with appropriate antithrombotic therapy, which plays a central role in all types of PAD. For years, antiplatelets have been indicated in patients with symptomatic PAD or those who have undergone revascularization. Unfortunately, a non-negligible proportion of patients with PAD will suffer from adverse events during the follow-up, even despite proper medical therapies for the prevention of PAD complications. Thus, there is room for improving clinical outcomes in these patients. Given the implication of both, primary and secondary hemostasis in arterial thrombosis and the pathophysiology of PAD, the combination of antiplatelets and anticoagulants has emerged as a potential antithrombotic alternative to antiplatelets alone. In this narrative review article, we have highlighted the most recent evidence about antithrombotic therapy in PAD patients, with a special focus on oral anticoagulation. Certainly, COMPASS and VOYAGER PAD trials have shown promising results. Thus, rivaroxaban in combination with aspirin seem to reduce cardiovascular outcomes with a similar bleeding risk compared to aspirin alone. Nevertheless, results from real-world studies are needed to confirm these observations, and other trials will provide novel evidence about the safety and efficacy of emerging anticoagulant agents.
Carfilzomib Promotes the Unfolded Protein Response and Apoptosis in Cetuximab-Resistant Colorectal Cancer
Authors: Zulkifli Ahmad , Tan Fiona H. , Areeb Zammam , Stuart Sarah F. , Gomez Juliana , Paradiso Lucia , Luwor Rodney B.
Published: July 2021

Cetuximab is a common treatment option for patients with wild-type K-Ras colorectal carcinoma. However, patients often display intrinsic resistance or acquire resistance to cetuximab following treatment. Here we generate two human CRC cells with acquired resistance to cetuximab that are derived from cetuximab-sensitive parental cell lines. These cetuximab-resistant cells display greater in vitro proliferation, colony formation and migration, and in vivo tumour growth compared with their parental counterparts. To evaluate potential alternative therapeutics to cetuximab-acquired-resistant cells, we tested the efficacy of 38 current FDA-approved agents against our cetuximab-acquired-resistant clones. We identified carfilzomib, a selective proteosome inhibitor to be most effective against our cell lines. Carfilzomib displayed potent antiproliferative effects, induced the unfolded protein response as determined by enhanced CHOP expression and ATF6 activity, and enhanced apoptosis as determined by enhanced caspase-3/7 activity. Overall, our results indicate a potentially novel indication for carfilzomib: that of a potential alternative agent to treat cetuximab-resistant colorectal cancer.
CMR-Based Risk Stratification of Sudden Cardiac Death and Use of Implantable Cardioverter–Defibrillator in Non-Ischemic Cardiomyopathy
Authors: Keil Laura , Chevalier Céleste , Kirchhof Paulus , Blankenberg Stefan , Lund Gunnar , Müllerleile Kai , Magnussen Christina
Published: July 2021

Non-ischemic cardiomyopathy (NICM) is one of the most important entities for arrhythmias and sudden cardiac death (SCD). Previous studies suggest a lower benefit of implantable cardioverter–defibrillator (ICD) therapy in patients with NICM as compared to ischemic cardiomyopathy (ICM). Nevertheless, current guidelines do not differentiate between the two subgroups in recommending ICD implantation. Hence, risk stratification is required to determine the subgroup of patients with NICM who will likely benefit from ICD therapy. Various predictors have been proposed, among others genetic mutations, left-ventricular ejection fraction (LVEF), left-ventricular end-diastolic volume (LVEDD), and T-wave alternans (TWA). In addition to these parameters, cardiovascular magnetic resonance imaging (CMR) has the potential to further improve risk stratification. CMR allows the comprehensive analysis of cardiac function and myocardial tissue composition. A range of CMR parameters have been associated with SCD. Applicable examples include late gadolinium enhancement (LGE), T1 relaxation times, and myocardial strain. This review evaluates the epidemiological aspects of SCD in NICM, the role of CMR for risk stratification, and resulting indications for ICD implantation.
Elastic Properties of Taurine Single Crystals Studied by Brillouin Spectroscopy
Authors: Kang Dong Hoon , Oh Soo Han , Ko Jae-Hyeon , Lee Kwang-Sei , Kojima Seiji
Published: July 2021

The inelastic interaction between the incident photons and acoustic phonons in the taurine single crystal was investigated by using Brillouin spectroscopy. Three acoustic phonons propagating along the crystallographic b-axis were investigated over a temperature range of −185 to 175 °C. The temperature dependences of the sound velocity, the acoustic absorption coefficient, and the elastic constants were determined for the first time. The elastic behaviors could be explained based on normal lattice anharmonicity. No evidence for the structural phase transition was observed, consistent with previous structural studies. The birefringence in the ac-plane indirectly estimated from the split longitudinal acoustic modes was consistent with one theoretical calculation by using the extrapolation of the measured dielectric functions in the infrared range.
In Silico Target Prediction of Overexpressed microRNAs from LPS-Challenged Zebrafish (Danio rerio) Treated with the Novel Anti-Inflammatory Peptide TnP
Authors: Disner Geonildo R. , Falcão Maria A. P. , Lima Carla , Lopes-Ferreira Monica
Published: July 2021

miRNAs regulate gene expression post-transcriptionally in various processes, e.g., immunity, development, and diseases. Since their experimental analysis is complex, in silico target prediction is important for directing investigations. TnP is a candidate peptide for anti-inflammatory therapy, first discovered in the venom of Thalassophryne nattereri, which led to miRNAs overexpression in LPS-inflamed zebrafish post-treatment. This work aimed to predict miR-21, miR-122, miR-731, and miR-26 targets using overlapped results of DIANA microT-CDS and TargetScanFish software. This study described 513 miRNAs targets using highly specific thresholds. Using Gene Ontology over-representation analysis, we identified their main roles in regulating gene expression, neurogenesis, DNA-binding, transcription regulation, immune system process, and inflammatory response. miRNAs act in post-transcriptional regulation, but we revealed that their targets are strongly related to expression regulation at the transcriptional level, e.g., transcription factors proteins. A few predicted genes participated concomitantly in many biological processes and molecular functions, such as foxo3a, rbpjb, rxrbb, tyrobp, hes6, zic5, smad1, e2f7, and npas4a. Others were particularly involved in innate immunity regulation: il17a/f2, pik3r3b, and nlrc6. Together, these findings not only provide new insights into the miRNAs mode of action but also raise hope for TnP therapy and may direct future experimental investigations.
Function of Circular RNAs in Fish and Their Potential Application as Biomarkers
Authors: Rbbani Golam , Nedoluzhko Artem , Galindo-Villegas Jorge , Fernandes Jorge M. O.
Published: July 2021

Circular RNAs (circRNAs) are an emerging class of regulatory RNAs with a covalently closed-loop structure formed during pre-mRNA splicing. Recent advances in high-throughput RNA sequencing and circRNA-specific computational tools have driven the development of novel approaches to their identification and functional characterization. CircRNAs are stable, developmentally regulated, and show tissue- and cell-type-specific expression across different taxonomic groups. They play a crucial role in regulating various biological processes at post-transcriptional and translational levels. However, the involvement of circRNAs in fish immunity has only recently been recognized. There is also broad evidence in mammals that the timely expression of circRNAs in muscle plays an essential role in growth regulation but our understanding of their expression and function in teleosts is still very limited. Here, we discuss the available knowledge about circRNAs and their role in growth and immunity in vertebrates from a comparative perspective, with emphasis on cultured teleost fish. We expect that the interest in teleost circRNAs will increase substantially soon, and we propose that they may be used as biomarkers for selective breeding of farmed fish, thus contributing to the sustainability of the aquaculture sector.
The Consequences of Soluble Epoxide Hydrolase Deletion on Tumorigenesis and Metastasis in a Mouse Model of Breast Cancer
Authors: Kesavan Rushendhiran , Frömel Timo , Zukunft Sven , Brüne Bernhard , Weigert Andreas , Wittig Ilka , Popp Rüdiger , Fleming Ingrid
Published: July 2021

Epoxides and diols of polyunsaturated fatty acids (PUFAs) are bioactive and can influence processes such as tumor cell proliferation and angiogenesis. Studies with inhibitors of the soluble epoxide hydrolase (sEH) in animals overexpressing cytochrome P450 enzymes or following the systemic administration of specific epoxides revealed a markedly increased incidence of tumor metastases. To determine whether PUFA epoxides increased metastases in a model of spontaneous breast cancer, sEH-/- mice were crossed onto the polyoma middle T oncogene (PyMT) background. We found that the deletion of the sEH accelerated the growth of primary tumors and increased both the tumor macrophage count and angiogenesis. There were small differences in the epoxide/diol content of tumors, particularly in epoxyoctadecamonoenic acid versus dihydroxyoctadecenoic acid, and marked changes in the expression of proteins linked with cell proliferation and metabolism. However, there was no consequence of sEH inhibition on the formation of metastases in the lymph node or lung. Taken together, our results confirm previous reports of increased tumor growth in animals lacking sEH but fail to substantiate reports of enhanced lymph node or pulmonary metastases.
Treating Seizures after Hypoxic-Ischemic Encephalopathy—Current Controversies and Future Directions
Authors: Zhou Kelly Q. , McDouall Alice , Drury Paul P. , Lear Christopher A. , Cho Kenta H. T. , Bennet Laura , Gunn Alistair J. , Davidson Joanne O.
Published: July 2021

Seizures are common in newborn infants with hypoxic-ischemic encephalopathy and are highly associated with adverse neurodevelopmental outcomes. The impact of seizure activity on the developing brain and the most effective way to manage these seizures remain surprisingly poorly understood, particularly in the era of therapeutic hypothermia. Critically, the extent to which seizures exacerbate brain injury or merely reflect the underlying evolution of injury is unclear. Current anticonvulsants, such as phenobarbital and phenytoin have poor efficacy and preclinical studies suggest that most anticonvulsants are associated with adverse effects on the developing brain. Levetiracetam seems to have less potential neurotoxic effects than other anticonvulsants but may not be more effective. Given that therapeutic hypothermia itself has significant anticonvulsant effects, randomized controlled trials of anticonvulsants combined with therapeutic hypothermia, are required to properly determine the safety and efficacy of these drugs. Small clinical studies suggest that prophylactic phenobarbital administration may improve neurodevelopmental outcomes compared to delayed administration; however, larger high-quality studies are required to confirm this. In conclusion, there is a distinct lack of high-quality evidence for whether and to what extent neonatal seizures exacerbate brain damage after hypoxia-ischemia and how best to manage them in the era of therapeutic hypothermia.
Effect of Lipopolysaccharide-Induced Inflammatory Challenge on β-Glucuronidase Activity and the Concentration of Quercetin and Its Metabolites in the Choroid Plexus, Blood Plasma and Cerebrospinal Fluid
Authors: Domżalska Małgorzata , Wiczkowski Wiesław , Szczepkowska Aleksandra , Chojnowska Sylwia , Misztal Tomasz , Walter Fruzsina R. , Deli Maria A. , Ishikawa Hiroshi , Schroten Horst , Schwerk Christian , Skipor Janina
Published: July 2021

Quercetin-3-glucuronide (Q3GA), the main phase II metabolite of quercetin (Q) in human plasma, is considered to be a more stable form of Q for transport with the bloodstream to tissues, where it can be potentially deconjugated by β-glucuronidase (β-Gluc) to Q aglycone, which easily enters the brain. This study evaluates the effect of lipopolysaccharide (LPS)-induced acute inflammation on β-Gluc gene expression in the choroid plexus (ChP) and its activity in blood plasma, ChP and cerebrospinal fluid (CSF), and the concentration of Q and its phase II metabolites in blood plasma and CSF. Studies were performed on saline- and LPS-treated adult ewes (n = 40) receiving Q3GA intravenously (n = 16) and on primary rat ChP epithelial cells and human ChP epithelial papilloma cells. We observed that acute inflammation stimulated β-Gluc activity in the ChP and blood plasma, but not in ChP epithelial cells and CSF, and did not affect Q and its phase II metabolite concentrations in plasma and CSF, except Q3GA, for which the plasma concentration was higher 30 min after administration (p < 0.05) in LPS- compared to saline-treated ewes. The lack of Q3GA deconjugation in the ChP observed under physiological and acute inflammatory conditions, however, does not exclude its possible role in the course of neurodegenerative diseases.