bluuebunny/crossref_metadata_2025 huggingface dataset:
Factors Influencing Public Participation in Community Disaster Mitigation Activities: A Comparison of Model and Nonmodel Disaster Mitigation Communities
Authors: Que Ting , Wu Yuxin , Hu Shiyu , Cai Jianmin , Jiang Nan , Xing Huige
Published: September 2022

Public participation in community-organized disaster mitigation activities is important for improving disaster mitigation capacity. With data from 260 questionnaires, this study compared the current status of public participation in model disaster mitigation communities and nonmodel communities in a geological-disaster-prone area. Three community-organized disaster mitigation education activities were compared cross-sectionally. A binary logistic regression was used to analyze the effects of attitude, perceived behavioral control, disaster experience, and other key factors on the public’s choice to participate in community disaster mitigation activities. The analysis results indicated that model communities had higher public participation in two efforts, evacuation drills and self-help skills training, and lower participation in activities that invited them to express their feedback than nonmodel communities. The influence of attitudinal factors on the decision to participate in disaster mitigation activities had a high similarity across community types. The public participation in model disaster mitigation communities is influenced by factors such as subjective norms and participation cognition; the behavior of people in nonmodel communities is influenced by factors such as previous experience with disasters, perceived behavioral control, risk perception, and participation cognition and has a greater potential for disaster mitigation community construction. This study provides practical evidence and theoretical support for strengthening the sustainable development of disaster mitigation community building.
Can Inbound Tourism Improve Regional Ecological Efficiency? An Empirical Analysis from China
Authors: Zhao Liang , Xu Lifei , Li Ling , Hu Jing , Mu Lin
Published: September 2022

Inbound tourism has an important impact on regional eco-efficiency. This paper uses the panel data of 31 provincial administrative units in China from 2005 to 2019; uses the improved DEA model to measure the regional ecological efficiency; and uses the panel threshold model to investigate input, output, and efficiency from the perspective of green technology innovation. Then, it explores the heterogeneous effects of inbound tourism on ecological efficiency. This paper finds that cross-border tourism has a positive impact on the ecological efficiency of tourist destinations. However, the degree of influence varies and will be changed with the level of regional green innovation. The main conclusions are as follows: (1) From an overall perspective, inbound tourism has a significant positive effect on ecological efficiency. (2) With the increase in green innovation investment and output, the promotion effect of inbound tourism on regional ecological efficiency first increases and then decreases. (3) The higher the green innovation efficiency, the greater the promotion effect of inbound tourism on ecological efficiency. Therefore, the Chinese government should encourage the development of inbound tourism, adopt greener innovative technologies that are cleaner and more environmentally friendly, and enhance the welfare effect of tourism on green economy.
OSH Performance within TQM Application in Construction Companies: A Qualitative Study in Saudi Arabia
Authors: Alghaseb Mohammed , Alshmlani Tariq
Published: September 2022

OSH plays a significant role in construction project success. Therefore, the aim of this study is to explore the influence of total quality management (TQM) application in improving occupational safety and health (OSH) within the context of Saudi construction companies. Factors were identified from structured literature reviews of previous relevant empirical studies. Then, these factors were theoretically framed into the concept of a triple bottom line (TBL), which includes three main dimensions: social, environmental, and economic. Thus, a semistructured interview survey was used to investigate these factors to address the performance of OSH in construction companies that implement TQM. A grounded theory was used to analyze and determine these factors. Accordingly, fourteen effective factors are identified. The survey findings indicate that the most influenced factors are the control of occupational accidents/injuries, the enhancement of workforce safety, the improvement of management pledges toward OSH, and the development of work culture toward OSH. These findings are vital in exploring the influence of TQM application in Saudi construction companies for the management of improving the performance of OSH, thereby helping to reduce the level of work injuries in the construction field and boosting the safety and health of workers for construction projects.
Sexist Attitudes in Adolescents: Prevalence and Associated Factors
Authors: Vila-Cortavitarte Elena , Díaz-Gómez N. Marta , Díaz-Gómez José Miguel
Published: September 2022

Gender violence is a major public health issue. The aim of this study was to determine the prevalence of sexist attitudes that could be associated with said violence, and to identify some sociodemographic variables that predict sexism. A cross-sectional observational study was conducted with 723 adolescents between the ages of 14 to 19. Their explicit sexist attitudes were measured with the EVAMVE and EARG scales, and their implicit attitudes were measured with a series of assessment items regarding the behavior of the protagonists of a video and a story in which a young couple interacts. Explicit and implicit sexist attitudes were detected in adolescents of both sexes. Qualitatively, the assessment of the behavior of the female protagonist is striking. Regarding the sexism predictors, it was found that male adolescents, those born outside of Spain, those who were studying in a public school, those whose parents did not have university studies, and those who consumed the most pornography presented attitudes that were significantly more sexist. These results suggest that it is necessary to strengthen education in equality and prevention of gender violence in adolescents, and to develop affective-sexual education programs.
Cultural Adaptation and Validation of the Premature Infant Pain Profile-Revised (PIPP-R) Pain Measurement Scale: Research Protocol
Authors: Núñez-López Irene , Collados-Gómez Laura , Abalo Raquel , Martínez-Pérez Patricia , Moreno-Vicente Álvaro , Cid-Expósito María-Gema
Published: September 2022

Introduction: The main objective of this study is to validate the PIPP-R scale (Premature Infant Pain Profile-Revised) for measuring neonatal pain in the Spanish hospital setting. Materials and Methods: The original scale will be translated from English into Spanish and a consensus translation will be prepared by the research team, which will be back-translated from Spanish into English. The content validity of the Spanish version of the scale will be measured using the Delphi method. Subsequently, a multicenter observational study will be conducted to assess construct validity, internal consistency, and intra-observer and inter-observer agreement. Pain will be assessed by comparing scores for a specific non-painful procedure with those for a specific painful procedure. The sample will include 300 subjects in intensive care and intermediate care units, who will be equally distributed among the participating hospitals. The subjects will be stratified into three groups by gestational age. Discussion: The original version of the PIPP-R scale is useful for objectively assessing neonatal acute and procedural pain from a gestational age of 25 weeks and over. It is important to culturally adapt the original validated scale and to test its validity and reliability in the Spanish healthcare context. The results of this study may represent significant progress in pain management.
Understanding the Non-Surgical Treatment Experience of Female Patients with Carpal Tunnel Syndrome: A Qualitative Study
Authors: Moro-López-Menchero Paloma , García-Bravo Cristina , Fernández-de-las-Peñas César , Güeita-Rodríguez Javier , Jiménez-Antona Carmen , Palacios-Ceña Domingo
Published: September 2022

Carpal tunnel syndrome (CTS) is a peripheral neuropathy of the upper extremity, characterized by pain, loss of strength, and decreased fine motor function. This study describes the experiences of women with CTS who received non-surgical treatments. A qualitative phenomenological study was undertaken. Purposive sampling was used. Women with clinical and electromyographic diagnoses of CTS were included. Eighteen in-depth interviews were conducted among women with CTS, and field notes were kept. The Giorgi’s approach was used for qualitative analysis of the data collected. Five themes emerged: (a) Seeking help and waiting for a diagnosis, (b) trying non-surgical therapeutic options, (c) avoiding invasive options, (d) treatment expectations, and (e) relationships with clinicians. The women described how diagnoses were delayed because women delay seeking help and referrals to medical specialists. Women avoid surgical options and prefer to opt for more conservative approaches, such as splinting or physical therapy. The main reason for avoiding surgical treatment is the fear of limitations and that surgery will not fully eliminate the symptoms. Conflicts may arise in the relationship with the clinician, and they demand to be able to participate in the decision-making process.
Examining the Indirect Death Surveillance System of The Great East Japan Earthquake and Tsunami
Authors: Zheng Xiang , Feng Chuyao , Ishiwatari Mikio
Published: September 2022

The long-term mortality risk of natural disasters is a key threat to disaster resilience improvement, yet an authoritative certification and a reliable surveillance system are, unfortunately, yet to be established in many countries. This study aimed to clarify the mechanism of post-disaster indirect deaths in Japan, to improve the existing disaster recovery evaluation system and support decision making in public policy. This study first investigated the definition of indirect deaths via a literature review before examining the observed number of indirect deaths via case study, census data from the Population Demographic and Household Surveys, other social surveys, and reports in the case of the Great East Japan Earthquake and Tsunami, which severely damaged northeastern Japan, especially the three prefectures, which are the target areas in this context (i.e., Fukushima, Iwate, and Miyagi). It was found that the reported number of indirect deaths was significantly underestimated. In total, 4657 indirect deaths were estimated to have occurred in the target prefectures. This was higher than the reported number, which was 3784. The overall statistics established via collaboration between local administrations and governments can be improved to provide better reference for researchers and policymakers to investigate the long-term effects of natural disaster.
The Structural Effects of Phosphorylation of Protein Arginine Methyltransferase 5 on Its Binding to Histone H4
Authors: Börzsei Rita , Bayarsaikhan Bayartsetseg , Zsidó Balázs Zoltán , Lontay Beáta , Hetényi Csaba
Published: September 2022

The protein arginine methyltransferase 5 (PRMT5) enzyme is responsible for arginine methylation on various proteins, including histone H4. PRMT5 is a promising drug target, playing a role in the pathomechanism of several diseases, especially in the progression of certain types of cancer. It was recently proved that the phosphorylation of PRMT5 on T80 residue increases its methyltransferase activity; furthermore, elevated levels of the enzyme were measured in the case of human hepatocellular carcinoma and other types of tumours. In this study, we constructed the complexes of the unmodified human PRMT5-methylosome protein 50 (MEP50) structure and its T80-phosphorylated variant in complex with the full-length histone H4 peptide. The full-length histone H4 was built in situ into the human PRMT5-MEP50 enzyme using experimental H4 fragments. Extensive molecular dynamic simulations and structure and energy analyses were performed for the complexed and apo protein partners, as well. Our results provided an atomic level explanation for two important experimental findings: (1) the increased methyltransferase activity of the phosphorylated PRMT5 when compared to the unmodified type; (2) the PRMT5 methylates only the free form of histone H4 not bound in the nucleosome. The atomic level complex structure H4-PRMT5-MEP50 will help the design of new inhibitors and in uncovering further structure–function relationships of PRMT enzymes.
Deep-Learning to Predict BRCA Mutation and Survival from Digital H&E Slides of Epithelial Ovarian Cancer
Authors: Nero Camilla , Boldrini Luca , Lenkowicz Jacopo , Giudice Maria Teresa , Piermattei Alessia , Inzani Frediano , Pasciuto Tina , Minucci Angelo , Fagotti Anna , Zannoni Gianfranco , Valentini Vincenzo , Scambia Giovanni
Published: September 2022

BRCA 1/2 genes mutation status can already determine the therapeutic algorithm of high grade serous ovarian cancer patients. Nevertheless, its assessment is not sufficient to identify all patients with genomic instability, since BRCA 1/2 mutations are only the most well-known mechanisms of homologous recombination deficiency (HR-d) pathway, and patients displaying HR-d behave similarly to BRCA mutated patients. HRd assessment can be challenging and is progressively overcoming BRCA testing not only for prognostic information but more importantly for drugs prescriptions. However, HR testing is not already integrated in clinical practice, it is quite expensive and it is not refundable in many countries. Selecting patients who are more likely to benefit from this assessment (BRCA 1/2 WT patients) at an early stage of the diagnostic process, would allow an optimization of genomic profiling resources. In this study, we sought to explore whether somatic BRCA1/2 genes status can be predicted using computational pathology from standard hematoxylin and eosin histology. In detail, we adopted a publicly available, deep-learning-based weakly supervised method that uses attention-based learning to automatically identify sub regions of high diagnostic value to accurately classify the whole slide (CLAM). The same model was also tested for progression free survival (PFS) prediction. The model was tested on a cohort of 664 (training set: n = 464, testing set: n = 132) ovarian cancer patients, of whom 233 (35.1%) had a somatic BRCA 1/2 mutation. An area under the curve of 0.7 and 0.55 was achieved in the training and testing set respectively. The model was then further refined by manually identifying areas of interest in half of the cases. 198 images were used for training (126/72) and 87 images for validation (55/32). The model reached a zero classification error on the training set, but the performance was 0.59 in terms of validation ROC AUC, with a 0.57 validation accuracy. Finally, when applied to predict PFS, the model achieved an AUC of 0.71, with a negative predictive value of 0.69, and a positive predictive value of 0.75. Based on these analyses, we have planned further steps of development such as proving a reference classification performance, exploring the hyperparameters space for training optimization, eventually tweaking the learning algorithms and the neural networks architecture for better suiting this specific task. These actions may allow the model to improve performances for all the considered outcomes.
Damaged DNA Is an Early Event of Neurodegeneration in Induced Pluripotent Stem Cell-Derived Motoneurons with UBQLN2P497H Mutation
Authors: Zhang Yiti , Zeng Baitao , Gu Ao , Kang Qinyu , Zhao Mingri , Peng Guangnan , Zhou Miaojin , Liu Wanxi , Liu Min , Ding Lingjie , Liang Desheng , Liu Xionghao , Liu Mujun
Published: September 2022

Ubiquilin-2 (UBQLN2) mutations lead to familial amyotrophic lateral sclerosis (FALS)/and frontotemporal dementia (FTLD) through unknown mechanisms. The combination of iPSC technology and CRISPR-mediated genome editing technology can generate an iPSC-derived motor neuron (iPSC-MN) model with disease-relevant mutations, which results in increased opportunities for disease mechanism research and drug screening. In this study, we introduced a UBQLN2-P497H mutation into a healthy control iPSC line using CRISPR/Cas9, and differentiated into MNs to study the pathology of UBQLN2-related ALS. Our in vitro MN model faithfully recapitulated specific aspects of the disease, including MN apoptosis. Under sodium arsenite (SA) treatment, we found differences in the number and the size of UBQLN2+ inclusions in UBQLN2P497H MNs and wild-type (WT) MNs. We also observed cytoplasmic TAR DNA-binding protein (TARDBP, also known as TDP-43) aggregates in UBQLN2P497H MNs, but not in WT MNs, as well as the recruitment of TDP-43 into stress granules (SGs) upon SA treatment. We noted that UBQLN2-P497H mutation induced MNs DNA damage, which is an early event in UBQLN2-ALS. Additionally, DNA damage led to an increase in compensation for FUS, whereas UBQLN2-P497H mutation impaired this function. Therefore, FUS may be involved in DNA damage repair signaling.
Reviewing the Modern Therapeutical Options and the Outcomes of Sacubitril/Valsartan in Heart Failure
Authors: Iovanovici Diana-Carina , Bungau Simona Gabriela , Vesa Cosmin Mihai , Moisi Madalina , Babes Elena Emilia , Tit Delia Mirela , Horvath Tunde , Behl Tapan , Rus Marius
Published: September 2022

Sacubitril/valsartan (S/V) is a pharmaceutical strategy that increases natriuretic peptide levels by inhibiting neprilysin and regulating the renin-angiotensin-aldosterone pathway, blocking AT1 receptors. The data for this innovative medication are mainly based on the PARADIGM-HF study, which included heart failure with reduced ejection fraction (HFrEF)-diagnosed patients and indicated a major improvement in morbidity and mortality when S/V is administrated compared to enalapril. A large part of the observed favorable results is related to significant reverse cardiac remodeling confirmed in two prospective trials, PROVE-HF and EVALUATE-HF. Furthermore, according to a subgroup analysis from the PARAGON-HF research, S/V shows benefits in HFrEF and in many subjects having preserved ejection fraction (HFpEF), which indicated a decrease in HF hospitalizations among those with a left ventricular ejection fraction (LVEF) < 57%. This review examines the proven benefits of S/V and highlights continuing research in treating individuals with varied HF characteristics. The article analyses published data regarding both the safeness and efficacy of S/V in patients with HF, including decreases in mortality and hospitalization, increased quality of life, and reversible heart remodeling. These benefits led to the HF guidelines recommendations updating and inclusion of S/V combinations a key component of HFrEF treatment.
CXCL8, CCL2, and CMV Seropositivity as New Prognostic Factors for a Severe COVID-19 Course
Authors: Pius-Sadowska Ewa , Niedźwiedź Anna , Kulig Piotr , Baumert Bartłomiej , Sobuś Anna , Rogińska Dorota , Łuczkowska Karolina , Ulańczyk Zofia , Wnęk Szymon , Karolak Igor , Paczkowska Edyta , Kotfis Katarzyna , Kawa Miłosz , Stecewicz Iwona , Zawodny Piotr , Machaliński Bogusław
Published: September 2022

The exact pathophysiology of severe COVID-19 is not entirely elucidated, but it has been established that hyperinflammatory responses and cytokine storms play important roles. The aim of this study was to examine CMV status, select chemokines, and complement components in COVID-19, and how concentrations of given molecules differ over time at both molecular and proteomic levels. A total of 210 COVID-19 patients (50 ICU and 160 non-ICU patients) and 80 healthy controls were enrolled in this study. Concentrations of select chemokines (CXCL8, CXCL10, CCL2, CCL3, CCR1) and complement factors (C2, C9, CFD, C4BPA, C5AR1, CR1) were examined at mRNA and protein levels with regard to a COVID-19 course (ICU vs. non-ICU group) and CMV status at different time intervals. We detected several significant differences in chemokines and complement profiles between ICU and non-ICU groups. Pro-inflammatory chemokines and the complement system appeared to greatly contribute to the pathogenesis and development of severe COVID-19. Higher concentrations of CXCL8 and CCL2 in the plasma, with reduced mRNA expression presumably through negative feedback mechanisms, as well as CMV-positive status, correlated with more severe courses of COVID-19. Therefore, CXCL8, CCL2, and CMV seropositivity should be considered as new prognostic factors for severe COVID-19 courses. However, more in-depth research is needed.
Melatonin Type 2 Receptor Activation Regulates Blue Light Exposure-Induced Mouse Corneal Epithelial Damage by Modulating Impaired Autophagy and Apoptosis
Authors: Jin Rujun , Li Ying , Jin Hui , Yoon Hee Su , Choi Ji Suk , Kim Jonghwa , Yoon Hyeon Jeong , Yoon Kyung Chul
Published: September 2022

The MT1/2 receptors, members of the melatonin receptor, belong to G protein-coupled receptors and mainly regulate circadian rhythms and sleep in the brain. Previous studies have shown that in many other cells and tissues, such as HEK293T cells and the retina, MT1/2 receptors can be involved in mitochondrial homeostasis, antioxidant, and anti-inflammatory responses. In our study, we aimed to investigate the effects of blue light (BL) exposure on the expression of melatonin and its receptors in the mouse cornea and to evaluate their functional role in corneal epithelial damage. After exposing 8-week-old C57BL/6 mice to BL at 25 and 100 J/cm2 twice a day for 14 days, a significant increase in the expression of 4-HNE and MT2 was observed in the cornea. MT2 antagonist-treated mice exposed to BL showed an increased expression of p62 and decreased expression of BAX and cleaved caspase 3 compared with mice exposed only to BL. In addition, MT2 antagonist-treated mice showed more enhanced MDA and corneal damage. In conclusion, BL exposure can induce MT2 expression in the mouse cornea. MT2 activation can modulate impaired autophagy and apoptosis by increasing the expression of BAX, an apoptosis activator, thereby regulating the progression of corneal epithelial damage induced by BL exposure.
Motor Behavioral Deficits in the Cuprizone Model: Validity of the Rotarod Test Paradigm
Authors: Lubrich Concordia , Giesler Paula , Kipp Markus
Published: September 2022

Multiple Sclerosis (MS) is a neuroinflammatory disorder, which is histopathologically characterized by multifocal inflammatory demyelinating lesions affecting both the central nervous system’s white and grey matter. Especially during the progressive phases of the disease, immunomodulatory treatment strategies lose their effectiveness. To develop novel progressive MS treatment options, pre-clinical animal models are indispensable. Among the various different models, the cuprizone de- and remyelination model is frequently used. While most studies determine tissue damage and repair at the histological and ultrastructural level, functional readouts are less commonly applied. Among the various overt functional deficits, gait and coordination abnormalities are commonly observed in MS patients. Motor behavior is mediated by a complex neural network that originates in the cortex and terminates in the skeletal muscles. Several methods exist to determine gait abnormalities in small rodents, including the rotarod testing paradigm. In this review article, we provide an overview of the validity and characteristics of the rotarod test in cuprizone-intoxicated mice.
Unraveling Mitochondrial Determinants of Tumor Response to Radiation Therapy
Authors: Zaffaroni Mattia , Vincini Maria Giulia , Corrao Giulia , Marvaso Giulia , Pepa Matteo , Viglietto Giuseppe , Amodio Nicola , Jereczek-Fossa Barbara Alicja
Published: September 2022

Radiotherapy represents a highly targeted and efficient treatment choice in many cancer types, both with curative and palliative intents. Nevertheless, radioresistance, consisting in the adaptive response of the tumor to radiation-induced damage, represents a major clinical problem. A growing body of the literature suggests that mechanisms related to mitochondrial changes and metabolic remodeling might play a major role in radioresistance development. In this work, the main contributors to the acquired cellular radioresistance and their relation with mitochondrial changes in terms of reactive oxygen species, hypoxia, and epigenetic alterations have been discussed. We focused on recent findings pointing to a major role of mitochondria in response to radiotherapy, along with their implication in the mechanisms underlying radioresistance and radiosensitivity, and briefly summarized some of the recently proposed mitochondria-targeting strategies to overcome the radioresistant phenotype in cancer.
TXNIP Promotes Human NK Cell Development but Is Dispensable for NK Cell Functionality
Authors: Persyn Eva , Wahlen Sigrid , Kiekens Laura , Taveirne Sylvie , Van Loocke Wouter , Van Ammel Els , Van Nieuwerburgh Filip , Taghon Tom , Vandekerckhove Bart , Van Vlierberghe Pieter , Leclercq Georges
Published: September 2022

The ability of natural killer (NK) cells to kill tumor cells without prior sensitization makes them a rising player in immunotherapy. Increased understanding of the development and functioning of NK cells will improve their clinical utilization. As opposed to murine NK cell development, human NK cell development is still less understood. Here, we studied the role of thioredoxin-interacting protein (TXNIP) in human NK cell differentiation by stable TXNIP knockdown or overexpression in cord blood hematopoietic stem cells, followed by in vitro NK cell differentiation. TXNIP overexpression only had marginal effects, indicating that endogenous TXNIP levels are sufficient in this process. TXNIP knockdown, however, reduced proliferation of early differentiation stages and greatly decreased NK cell numbers. Transcriptome analysis and experimental confirmation showed that reduced protein synthesis upon TXNIP knockdown likely caused this low proliferation. Contrary to its profound effects on the early differentiation stages, TXNIP knockdown led to limited alterations in NK cell phenotype, and it had no effect on NK cell cytotoxicity or cytokine production. Thus, TXNIP promotes human NK cell differentiation by affecting protein synthesis and proliferation of early NK cell differentiation stages, but it is redundant for functional NK cell maturation.
Multiple Timescale Dynamic Analysis of Functionally-Impairing Mutations in Human Ileal Bile Acid-Binding Protein
Authors: Horváth Gergő , Balterer Bence , Micsonai András , Kardos József , Toke Orsolya
Published: September 2022

Human ileal bile acid-binding protein (hI-BABP) has a key role in the enterohepatic circulation of bile salts. Its two internal binding sites exhibit positive cooperativity accompanied by a site-selectivity of glycocholate (GCA) and glycochenodeoxycholate (GCDA), the two most abundant bile salts in humans. To improve our understanding of the role of dynamics in ligand binding, we introduced functionally impairing single-residue mutations at two key regions of the protein and subjected the mutants to NMR relaxation analysis and MD simulations. According to our results, mutation in both the vicinity of the C/D (Q51A) and the G/H (Q99A) turns results in a redistribution of motional freedom in apo hI-BABP. Mutation Q51A, deteriorating the site-selectivity of GCA and GCDA, results in the channeling of ms fluctuations into faster motions in the binding pocket hampering the realization of key side chain interactions. Mutation Q99A, abolishing positive binding cooperativity for GCDA, leaves ms motions in the C-terminal half unchanged but by decoupling βD from a dynamic cluster of the N-terminal half displays an increased flexibility in the vicinity of site 1. MD simulations of the variants indicate structural differences in the portal region and mutation-induced changes in dynamics, which depend on the protonation state of histidines. A dynamic coupling between the EFGH portal, the C/D-region, and the helical cap is evidenced highlighting the interplay of structural and dynamic effects in bile salt recognition in hI-BABP.
Loss of Function of the RRMF Domain in OsROS1a Causes Sterility in Rice (Oryza sativa L.)
Authors: Xu Jian-Hong , Irshad Faiza , Yan Yan , Li Chao
Published: September 2022

For crop seed production, the development of anthers and male fertility are the main agronomic traits and key biological processes for flowering plants. Active DNA demethylation regulates many plant developmental processes and is ensured by 5-meC DNA glycosylase enzymes. To find out the role of OsROS1a, OsROS1a gene editing mutants were generated using the CRISPR/Cas9 system. The osros1a mutants had shrink spikelets, smaller anthers and pollen grains, and were not stained by iodine staining showing a significant reduction in total soluble sugar and starch contents as compared to wildtype (WT), which caused complete male sterility. Similarly, the expression of genes involved in pollen and anther development was decreased in osros1a mutants as compared to WT. Furthermore, bisulfite sequencing showed that the CG and CHG methylation of the OsPKS2 gene promoter was significantly increased in the osros1a mutant, which caused a reduced expression of OsPKS2 in osros1a mutants. DNA methylation of the TDR gene promoter was similar between WT and osros1a mutants, indicating that the DNA methylation effect by OsROS1a was gene specific. The expression of OsROS1a in the mutants was not changed, but it produced a frame-shift mutation to truncate the Pem-CXXC and RRMF domains. Combined with previous studies, our findings suggested that the RRMF domain in OsROS1a is the functional domain and loss of RRMF for OsROS1a causes sterility in rice.
Restoration of Mitochondrial Function Is Essential in the Endothelium-Dependent Vasodilation Induced by Acacetin in Hypertensive Rats
Authors: Li Yuan , Dang Qingya , Li Zhiyi , Han Chuting , Yang Yan , Li Miaoling , Li Pengyun
Published: September 2022

Mitochondrial dysfunction in the endothelium contributes to the progression of hypertension and plays an obligatory role in modulating vascular tone. Acacetin is a natural flavonoid compound that has been shown to possess multiple beneficial effects, including vasodilatation. However, whether acacetin could improve endothelial function in hypertension by protecting against mitochondria-dependent apoptosis remains to be determined. The mean arterial pressure (MAP) in Wistar Kyoto (WKY) rats, spontaneously hypertensive rats (SHR) administered with acacetin intraperitoneally for 2 h or intragastrically for six weeks were examined. The endothelial injury was evaluated by immunofluorescent staining and a transmission electron microscope (TEM). Vascular tension measurement was performed to assess the protective effect of acacetin on mesenteric arteries. Endothelial injury in the pathogenesis of SHR was modeled in HUVECs treated with Angiotensin II (Ang II). Mitochondria-dependent apoptosis, the opening of Mitochondrial Permeability Transition Pore (mPTP) and mitochondrial dynamics proteins were determined by fluorescence activated cell sorting (FACS), immunofluorescence staining and western blot. Acacetin administered intraperitoneally greatly reduced MAP in SHR by mediating a more pronounced endothelium-dependent dilatation in mesenteric arteries, and the vascular dilatation was reduced remarkably by NG-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NO synthesis. While acacetin administered intragastrically for six weeks had no apparent effect on MAP, it improved the endothelium-dependent dilatation in SHR by activating the AKT/eNOS pathway and protecting against the abnormalities of endothelium and mitochondria. Furthermore, acacetin remarkably inhibited Ang II induced apoptosis by inhibiting the increased expression of Cyclophilin D (CypD), promoted the opening of mPTP, ROS generation, ATP loss and disturbance of dynamin-related protein 1 (DRP1)/optic atrophy1 (OPA1) dynamics in HUVECs. This study suggests that acacetin protected against endothelial dysfunction in hypertension by activating the AKT/eNOS pathway and modulating mitochondrial function by targeting mPTP and DRP1/OPA1-dependent dynamics.
Topical Treatment of Actinic Keratosis and Metalloproteinase Expression: A Clinico-Pathological Retrospective Study
Authors: Campione Elena , Di Prete Monia , Di Raimondo Cosimo , Costanza Gaetana , Palumbo Vincenzo , Garofalo Virginia , Mazzilli Sara , Franceschini Chiara , Dika Emi , Bianchi Luca , Orlandi Augusto
Published: September 2022

Actinic keratosis is an intraepithelial proliferation of atypical keratinocytes that could progress into invasive squamous cell carcinoma. Most evidence suggests an important role of the dermal matrix metalloproteinases in the progression of atypical skin epithelial lesions. We evaluated the clinical efficacy of three different therapeutic modalities (a medical device containing 0.8% piroxicam cream and 50+ sunscreen, photodynamic therapy, and ingenol mebutate gel) to treat suspicious actinic keratoses, which were biopsied for histopathological examination and then analyzed for the expression of matrix metalloproteinases by immunohistochemistry. Clinical, dermoscopic, and reflectance confocal microscopy evaluations revealed a gradual decrease in all standard scores validated for actinic keratosis assessment at the end of the treatments. From a histopathological point of view, we documented the substantial restoration of normal skin architecture, while the immunohistochemical evaluation of matrix metalloproteinases showed a reduction in expression in the treated skin lesions compared to the baseline. As actinic keratoses are considered the precursors of squamous cell carcinoma, their treatment is crucial to prevent the development of a more aggressive disease. Our study monitored the evolution of actinic keratoses subjected to three different topical therapies, with the value of correlating clinical and histopathological findings. Moreover, as the matrix metalloproteinases are largely recognized factors involved in the pathogenesis and evolution of actinic keratosis to squamous cell carcinoma, the demonstration by immunohistochemistry of a reduction in their expression after the treatments adds new valuable concern to the field.
Dynamic Assessment of Tissue and Plasma EGFR-Activating and T790M Mutations with Droplet Digital PCR Assays for Monitoring Response and Resistance in Non-Small Cell Lung Cancers Treated with EGFR-TKIs
Authors: Ho Hsiang-Ling , Wang Fang-Yu , Chiang Chi-Lu , Tsai Chun-Ming , Chiu Chao-Hua , Chou Teh-Ying
Published: September 2022

Assessing tumor EGFR mutation status is necessary for the proper management of patients with advanced non–small cell lung cancer (NSCLC). We evaluated the impact of dynamic analyses of the plasma and tissue EGFR mutation using ultra-sensitive droplet digital PCR (ddPCR) assays to manage NSCLC patients treated with EGFR tyrosine kinase inhibitors (EGFR-TKIs). Paired tumor tissues and plasma samples from 137 EGFR-mutated lung adenocarcinoma patients prior to the first-line EGFR-TKIs treatment (at baseline) and at disease progression were subjected to EGFR mutation analysis using ddPCR, together with the analyses of the clinicopathological characteristics and treatment outcomes. Patients with EGFR-activating mutations detected in baseline plasma were associated with bone metastasis (p = 0.002) and had shorter progression-free survival (12.9 vs. 17.7 months, p = 0.02) and overall survival (24.0 vs. 39.4 months, p = 0.02) compared to those without. Pre-treatment EGFR T790M mutation found in baseline tumor tissues of 28 patients (20.4%; 28/137) was significantly associated with brain metastasis (p = 0.005) and a shorter brain metastasis-free survival (p = 0.001). The presence of EGFR T790M mutations in baseline tumor tissues did not correlate with the emergence of acquired EGFR T790M mutations detected at progression. At disease progression, acquired EGFR T790M mutations were detected in 26.6% (21/79) of the plasma samples and 42.9% (15/35) of the rebiopsy tissues, with a concordance rate of 71.4% (25/35). The dynamic monitoring of tissue and plasma EGFR mutation status at baseline and progression using ddPCR has a clinical impact on the evaluation of EGFR-TKIs treatment efficacy and patient outcomes, as well as the emergence of resistance in NSCLC.
An Analysis of the Effects of In Vitro Photodynamic Therapy on Prostate Cancer Tissue by Histopathological Examination and Magnetic Resonance Imaging
Authors: Aebisher David , Osuchowski Michał , Bartusik-Aebisher Dorota , Krupka-Olek Magdalena , Dynarowicz Klaudia , Kawczyk-Krupka Aleksandra
Published: September 2022

Prostate cancer can significantly shorten the lifetime of a patient, even if he is diagnosed at an early stage. The development of minimally-invasive focal therapies such as photodynamic therapy to reduce the number of neoplastic cells while sparing delicate structures is extremely advantageous for treating prostate cancer. This study investigates the effect of photodynamic therapy performed in prostate tissue samples in vitro, using quantitative magnetic resonance imaging and histopathological analysis. Prostate tissue samples were treated with oxygenated solutions of Rose Bengal (RB) or protoporphyrin IX disodium salt (PpIX), illuminated with visible light, and then analyzed for changes in morphology by microscopy and by measurement of spin–lattice and spin–spin relaxation times at 1.5 Tesla. In the treated prostate tissue samples, histopathological images revealed chromatin condensation and swelling of the stroma, and in some cases, thrombotic necrosis and swelling of the stroma accompanied by pyknotic nuclei occurred. Several samples had protein fragments in the stroma. Magnetic resonance imaging of the treated prostate tissue samples revealed differences in the spin–lattice and spin–spin relaxation times prior to and post photodynamic action.
Cellular Lactate Spectroscopy Using 1.5 Tesla Clinical Apparatus
Authors: Truszkiewicz Adrian , Bartusik-Aebisher Dorota , Zalejska-Fiolka Jolanta , Kawczyk-Krupka Aleksandra , Aebisher David
Published: September 2022

Cellular lactate is a key cellular metabolite and marker of anaerobic glycolysis. Cellular lactate uptake, release, production from glucose and glycogen, and interconversion with pyruvate are important determinants of cellular energy. It is known that lactate is present in the spectrum of neoplasms and low malignancy (without necrotic lesions). Also, the appearance of lactate signals is associated with anaerobic glucose, mitochondrial dysfunction, and other inflammatory responses. The aim of this study was the detection of lactate in cell cultures with the use of proton magnetic resonance (1H MRS) and a 1.5 Tesla clinical apparatus (MR OPTIMA 360), characterized as a medium-field system. In this study, selected metabolites, together with cellular lactate, were identified with the use of an appropriate protocol and management algorithm. This paper describes the results obtained for cancer cell cultures. This medium-field system has proven the possibility of detecting small molecules, such as lactate, with clinical instruments. 1H MRS performed using clinical MR apparatus is a useful tool for clinical analysis.
Genome-Wide Identification, Characterization and Expression Analysis of Toll-like Receptors in Marbled Rockfish (Sebastiscus marmoratus)
Authors: Zhang Yuan , Wang Xiaoyan , Han Fei , Gao Tianxiang
Published: September 2022

Toll-like receptor (TLR) is a cluster of type I transmembrane proteins that plays a role in innate immunity. Based on the marbled rockfish (Sebastiscus marmoratus) genome database, this study used bioinformatics methods to identify and analyze its TLR gene family members. The results showed that there were 11 TLR gene family members in Sebastiscus marmoratus (SmaTLR), which could be divided into five different subfamilies. The number of amino acids encoded by the Smatlr genes ranged from 637 to 1206. The physicochemical properties of the encoded proteins of different members were also computed. The results of protein structure prediction, phylogenetic relation, and motif analysis showed that the structure and function of the SmaTLRs were relatively conserved. Quantitative Real-Time PCR (qRT-PCR) analysis revealed the expression patterns of SmaTLRs in the gill, liver, spleen, head kidney, kidney, and intestine. SmaTLRs were widely detected in the tested tissues, and they tended to be expressed higher in immune-related tissues. After polyriboinosinic polyribocytidylic acid (poly(I:C)) challenge, SmaTLR14, SmaTLR3, SmaTLR5S, SmaTLR7, and SmaTLR22 were significantly upregulated in the spleen or liver. The results of this study will help to understand the status of TLR gene family members of marbled rockfish and provide a basis for further study of the functional analysis of this gene family.
Theoretical Explanation for the Rarity of Antibody-Dependent Enhancement of Infection (ADE) in COVID-19
Authors: Boldova Anna E. , Korobkin Julia D. , Nechipurenko Yury D. , Sveshnikova Anastasia N.
Published: September 2022

Global vaccination against the SARS-CoV-2 virus has proved to be highly effective. However, the possibility of antibody-dependent enhancement of infection (ADE) upon vaccination remains underinvestigated. Here, we aimed to theoretically determine conditions for the occurrence of ADE in COVID-19. We developed a series of mathematical models of antibody response: model Ab—a model of antibody formation; model Cv—a model of infection spread in the body; and a complete model, which combines the two others. The models describe experimental data on SARS-CoV and SARS-CoV-2 infections in humans and cell cultures, including viral load dynamics, seroconversion times and antibody concentration kinetics. The modelling revealed that a significant proportion of macrophages can become infected only if they bind antibodies with high probability. Thus, a high probability of macrophage infection and a sufficient amount of pre-existing antibodies are necessary for the development of ADE in SARS-CoV-2 infection. However, from the point of view of the dynamics of pneumocyte infection, the two cases where the body has a high concentration of preexisting antibodies and a high probability of macrophage infection and where there is a low concentration of antibodies in the body and no macrophage infection are indistinguishable. This conclusion could explain the lack of confirmed ADE cases for COVID-19.
Photochemical Consideration in the Interactions between Blood Proteins and Layered Inorganic Materials
Authors: Yamaguchi Tetsuo , Kim Hyoung-Mi , Oh Jae-Min
Published: September 2022

Interactions between layered double hydroxide (LDH) nanomaterials and plasma proteins according to their particle size and surface charge were evaluated. The LDHs with different particle size (150, 350 and 2000 nm) were prepared by adjusting hydrothermal treatment and urea hydrolysis and subsequent organic coating with citrate, malite and serite was applied to control the surface charge (ζ-potential: −15, 6 and 36 mV). Adsorption isotherms and Stern–Volmer plots for fluorescence quenching indicated that the human blood plasma had weak interactions toward all the types of LDHs. The adsorption isotherms did not show significant differences in the size and surface charges, while the fluorescence quenching ratio increased with the increase in the surface charge, implying that electrostatic interaction played a major role in their interactions. The fluorescence quenching of three types of plasma proteins (human serum albumin, γ-globulin and fibrinogen) by the surface charge-controlled LDHs suggested that the proteins adsorbed on the LDHs with a single layer and additional proteins were weakly adsorbed to surround the LDHs with adsorbed proteins. It was concluded that the LDH nanomaterials are fairly compatible for blood components due to the protein corona while the electrostatic interaction can affect their interaction with the proteins.
Gadolinium Accumulation and Toxicity on In Vitro Grown Stevia rebaudiana: A Case-Study on Gadobutrol
Authors: Scurtu Violeta Florina , Clapa Doina , Leopold Loredana Florina , Ranga Floricuța , Iancu Ștefania D. , Cadiș Adrian Ionuț , Coman Vasile , Socaci Sonia Ancuța , Moț Augustin C. , Coman Cristina
Published: September 2022

Gadolinium-based contrast agents are molecular complexes which are extensively used for diagnostic purposes. Apart from their tremendous contribution to disease diagnostics, there are several issues related to their use. They are extremely stable complexes and potential contaminants of surface and ground waters, an issue which is documented worldwide. The irrigation of fields with contaminated surface waters or their fertilization with sludge from wastewater treatment plants can lead to the introduction of Gd into the human food supply chain. Thus, this study focused on the potential toxicity of Gd on plants. For this purpose, we have studied the molecular effects of gadobutrol (a well-known MRI contrast agent) exposure on in vitro-grown Stevia rebaudiana. The effects of gadobutrol on plant morphology, on relevant plant metabolites such as chlorophylls, carotenoids, ascorbic acids (HPLC), minerals (ICP-OES), and on the generation of free radical species (MDA assay and EPR) were assessed. Exposures of 0.01, 0.05, 0.1, 1, and 3 mM gadobutrol were used. We found a correlation between the gadobutrol dose and the plant growth and concentration of metabolites. Above the 0.1. mM dose of gadobutrol, the toxic effects of Gd+3 ions became significant.
Metallophenolomics: A Novel Integrated Approach to Study Complexation of Plant Phenolics with Metal/Metalloid Ions
Authors: Fedenko Volodymyr S. , Landi Marco , Shemet Sergiy A.
Published: September 2022

Plant adaptive strategies have been shaped during evolutionary development in the constant interaction with a plethora of environmental factors, including the presence of metals/metalloids in the environment. Among adaptive reactions against either the excess of trace elements or toxic doses of non-essential elements, their complexation with molecular endogenous ligands, including phenolics, has received increasing attention. Currently, the complexation of phenolics with metal(loid)s is a topic of intensive studies in different scientific fields. In spite of the numerous studies on their chelating capacity, the systemic analysis of phenolics as plant ligands has not been performed yet. Such a systematizing can be performed based on the modern approach of metallomics as an integral biometal science, which in turn has been differentiated into subgroups according to the nature of the bioligands. In this regard, the present review summarizes phenolics–metal(loid)s’ interactions using the metallomic approach. Experimental results on the chelating activity of representative compounds from different phenolic subgroups in vitro and in vivo are systematized. General properties of phenolic ligands and specific properties of anthocyanins are revealed. The novel concept of metallophenolomics is proposed, as a ligand-oriented subgroup of metallomics, which is an integrated approach to study phenolics–metal(loid)s’ complexations. The research subjects of metallophenolomics are outlined according to the methodology of metallomic studies, including mission-oriented biometal sciences (environmental sciences, food sciences and nutrition, medicine, cosmetology, coloration technologies, chemical sciences, material sciences, solar cell sciences). Metallophenolomics opens new prospects to unite multidisciplinary investigations of phenolic–metal(loid) interactions.