bluuebunny/crossref_metadata_2025 huggingface dataset:
Dietary Sodium and Potassium Intake in Hungarian Elderly: Results from the Cross-Sectional Biomarker2019 Survey
Authors: Sarkadi-Nagy Eszter , Horváth Andrea , Varga Anita , Zámbó Leonóra , Török Andrea , Guba Georgina , Szilfai Nikolett , Zentai Andrea , Bakacs Márta
Published: August 2021

High sodium intake and inadequate potassium intake are associated with high blood pressure. The elderly are more salt sensitive than other age groups, yet a reliable estimate of the dietary sodium and potassium intake of this age group in Hungary is unavailable. The study aimed to estimate the sodium and potassium intakes in the Hungarian elderly from 24 h urine sodium and potassium excretion. In this cross-sectional study, participants were selected from patients of general practitioners practicing in western Hungary. The participants comprised 99 men and 90 women (mean age 67.1 (SD 5.4] years) who participated in the Biomarker2019 survey and returned a complete 24 h urine collection. We assessed dietary sodium and potassium by determining 24 h urinary sodium and potassium excretions and 3-day dietary records. The mean urinary sodium was 188.8 (73.5) mmoL/day, which is equivalent to 11.0 g of salt/day; and the mean urinary potassium was 65.8 (24.3) mmoL/day, which is equivalent to 3.03 g of potassium/day, after adjusting for non-urinary potassium losses. Only 7% of the subjects met the World Health Organization’s recommended target of less than 5 g of salt/day, and 33% consumed at least the recommended potassium amount of 3.5 g/day, based on the estimates from 24 h urine excretion. For most elderly, sodium intake exceeds, and potassium does not reach, dietary recommendations. The results underline the need to intensify salt reduction efforts in Hungary.
Effect of Mobile-Based Lifestyle Intervention on Weight Loss among the Overweight and Obese Elderly Population in China: A Randomized Controlled Trial
Authors: Zhou Mingzhu , Zhang Na , Zhang Yu , Yan Xinyu , Li Muxia , Guo Wen , Guo Xiaohui , He Hairong , Guo Kaiwei , Ma Guansheng
Published: August 2021

Background and Objective: Overweight or obesity, as an independent risk factor for chronic diseases, has been on the rise globally. Adopting a healthy lifestyle is positive to weight control. Mobile-based lifestyle interventions have shown potential benefits in weight loss, but most studies were carried out among non-elderly population, so it is necessary to perform well-designed randomized controlled trials among the elderly with overweight or obesity. The purpose of this study is to assess the effect of mobile-based lifestyle intervention on weight loss among the overweight and obese elderly population in China. Methods: This is a prospective, open-labeled, three-month, multicenter, randomized controlled trial involving 750 participants from five cities who were randomly assigned to dietary and physical activity interventions group (DPG; mobile phone with the App and bracelet), physical activity interventions group (PG; mobile phone with the App and bracelet) and control group (CG; no interventions and kept their lifestyle as before). The outcomes evaluated were changes in weight, body mass index (BMI), waist circumference (WC), and hip circumference (HC). Results: In total, 642 (85.6%) participants completed the study, 237 (94.8%), 203 (81.2%), and 202 (80.8%) for DPG, PG, and CG respectively. Comparing with PG and CG, the DPG showed a significant decrease in all outcomes after three months, including body weight (−4.1 kg vs. −1.0 kg; −4.1 kg vs. −0.8 kg; p < 0.05), BMI (−1.6 kg/m2 vs. −0.4 kg/m2; −1.6 kg/m2 vs. −0.3 kg/m2; p < 0.05), WC (−2.8 cm vs. −0.1 cm; −2.8 cm vs. −0.5 cm; p < 0.05), and HC (−3.8 cm vs. −1.3 cm; −3.8 cm vs. −1.3 cm; p < 0.05). Similar effects were seen across sex and BMI subgroups. Conclusions: Mobile-based lifestyle intervention obtained beneficial effect in weight loss among the elderly with overweight or obesity. Nevertheless, further studies are needed to confirm the effectiveness and its sustainability.
Induction of SARS-CoV-2-Specific IgG and IgA in Serum and Milk with Different SARS-CoV-2 Vaccines in Breastfeeding Women: A Cross-Sectional Study in Northern Spain
Authors: Lechosa-Muñiz Carolina , Paz-Zulueta María , Mendez-Legaza Jose Manuel , Irure-Ventura Juan , Cuesta González Rocío , Calvo Montes Jorge , López-Hoyos Marcos , Llorca Javier , Cabero-Pérez María Jesús
Published: August 2021

Breastfeeding mothers were excluded from the clinical trials conducted for vaccines against SARS-CoV-2. Since the start of the vaccination, some doubts have arisen regarding its compatibility with breastfeeding. The aim of this study was to analyse the presence of anti-SARS-CoV-2 antibodies in breast milk and serum (IgG and IgA) of vaccinated breastfeeding women. The main variables of the observational study were: adverse related events after vaccination and determination of the presence of IgG and IgA isotypes antibodies in serum and in breast milk of vaccinated women against the SARS-CoV-2 antigens. Results: 110 breastfeeding mothers were included; 70 women (63.6%) were vaccinated with two doses of BNT162b2, 20 women (18.2%) with two doses of mRNA-1273, and 20 women (18.2%) with a single dose of ChAdOx1-S. Regarding adverse reactions and vaccine safety, 38 women had no adverse reactions; 20 (18.2%) had general malaise or adenopathies; 10 (9.1%) had a headache; and 7 (6.4%) had fever. When analysing IgG antibodies, significantly higher levels of antibodies were found in serum and breast milk from mothers vaccinated with BNT162b2 or mRNA-1273 vs. ChAdOx1-S (p < 0.001 and p = 0.001, respectively). Analysing IgA antibodies, significant differences were found when comparing mean values in serum from mothers vaccinated with BNT162b2 or mRNA-1273 vs. ChAdOx1-S (0.12, 0.16, and 0.02, respectively; p < 0.001) and breast milk of mothers vaccinated when comparing BNT16b2 vs. ChAdOx1-S. All vaccinated breastfeeding mothers had serum anti-S1 IgG antibodies in response to vaccination against SARS-CoV-2, regardless of the commercial vaccine administered. Conclusions: the anti-SARS-CoV-2 vaccines were well tolerated by the mothers and the breastfed infant. In addition, breastfeeding mothers offer their infants IgA and IgG isotype antibodies directed against SARS-CoV-2 protein S in breast milk.
The Effects of City-County Mergers on Urban Energy Intensity: Empirical Evidence from Chinese Cities
Authors: Li Zhi , Yan Ruyi , Zhang Zuo , Sun Yue , Zhang Xiaogang
Published: August 2021

The adjustment of administrative division is one of the powerful tools used to promote urbanization by the Chinese government in recent decades, and there is little literature to discuss whether the government-led expansion of urban space through city-county mergers can bring about a decline in urban energy intensity. With the multi-period difference-in-differences (DID) method and comprehensive urban datasets, this research investigates the urban energy intensity results of the city-county mergers policy experiment in China from 2000 to 2017. We present evidence that city-county mergers are indeed beneficial in reducing urban energy intensity, and that the energy-saving effect of the policy only starts to become significant in the third year after implementation. We also further adopt a series of robustness tests, such as the counterfactual test, placebo tests and PSM-DID tests to find if this effect still exists. The mechanism test with mediating effects indicates they are potential contributors to the positive effects of mergers with moderate fiscal centralization, population agglomeration and regional integration. We further explore positive effects of mergers, relying on the scientifically design official appraisal system and improve government efficiency.
A Critical Review of EU Key Indicators for the Transition to the Circular Economy
Authors: Pacurariu Roxana Lavinia , Vatca Sorin Daniel , Lakatos Elena Simina , Bacali Laura , Vlad Mircea
Published: August 2021

The objective of this paper is to analyze the extent to which the system of indicators that is used in the Monitoring Framework for the transition to the circular economy (CE) is efficient and relevant in their contribution to the sustainable development of European communities. The fundaments of the transition framework and the main characteristics of the circularity indicators are presented. A critical review was performed in order to fulfill the objective of analyzing the current indicators. It is concluded that the indicators in the current framework are (as a selection from a very broad range of indicators theoretically proposed and with estimated practical applicability) limited from the perspective of circularity only to waste generation and recycling processes containing recyclable materials, without including important circularity indicators related to the prolonging and extending the life cycle of products and materials. This paper proposes and defines such an indicator, based on the consideration of the fundamental scalars describing economy, mass, energy, time, and value, respectively. The indicator is described and its applicability in all the phases of the economy is estimated.
Exploring Shank Circumference by Stretching after Training among Volleyball Players
Authors: Chen Yi-Lang , Tsai Fang-Min , Hsu Wei-Chen , Yang Chun-Ju , Yei Ting-Yo
Published: August 2021

This preliminary study examined the effects of a stretching intervention after training and its duration (15 vs. 30 min) on participants’ shank circumference (SC) reduction and subjective discomfort score. Ten male volleyball players underwent a routine 3 h training. A two-way analysis of variance revealed that the stretching intervention had significant effects on SC reduction (p < 0.01) and subjective discomfort scores (p < 0.001). Stretching after training could help eliminate shank strain, and a slighter discomfort in shanks when stretching was also seen (score, 20.1/100). An independent-samples t test revealed a significantly higher SC reduction (p < 0.01) with 30 min of stretching (5.6 mm) than with 15 min of stretching (2.7 mm); both stretching durations reduced SC significantly more than the no-stretching condition did. The findings of this study can serve as a reference for volleyball players to alleviate shank strain after daily routine training.
Heated Tobacco Products and Nicotine Pouches: A Survey of People with Experience of Smoking and/or Vaping in the UK
Authors: Brose Leonie S. , McDermott Máirtín S. , McNeill Ann
Published: August 2021

Background: To gauge the public health impact of new nicotine products, information is needed on use among different populations. Aims were to assess in adults who smoked, vaped, did both or had recently stopped: (1) awareness, ever and current use of heated tobacco products (HTPs) and nicotine pouches (NP), (2) characteristics associated with ever use, (3) reasons for use of and satisfaction with HTPs, (4) characteristics associated with interest in use of HTPs. Methods: Online survey in the UK in 2019, n = 3883. (1) Proportion aware, ever and current (≥monthly) use; (2) ever use regressed onto socio-demographics and smoking/vaping; (3) frequency of reasons for HTP use and satisfaction; (4) interest in trying HTPs regressed onto socio-demographics and smoking/vaping status. Results: Awareness was 34.8% for HTP and 15.9% for NP; current use was 3.2% and 2.7%. Being <45 years, higher education, living in London and currently both smoking and vaping were associated with ever having used the products. Curiosity was the most common reason for HTP use (79.8%) and 72.0% of ever HTP users found them at least as satisfying as smoking. Among those not currently using HTPs, 48.5% expressed any interest—lower among those aged over 65 and higher among those smoking and vaping. Conclusions: In this sample of adults with a history of nicotine use, very few currently used heated tobacco products or nicotine pouches. Satisfaction with and interest in HTPs were substantial. The low level of use is unlikely to substantially reduce the public health impact of smoking.
Health, Identification and Pleasure: An Ethnographic Study on the Self-Management and Construction of Taijiquan Park Culture Space
Authors: Ma Xiujie , Xie Jing , Jennings George
Published: August 2021

The public space of a park is one of the most important carriers of social interaction and cultural practice in urban areas. Taking an ethnography of Taijiquan in Chengdu (China) as a case study, this article explores the production of Taijiquan’s “park culture space” (PCS). Our analysis revealed that the development of PCS not only transformed “public space” in the park to a “private space” through Taijiquan practice and exchange but also transformed “material space” in the park into “social space” with identification. We found that working on the process of self-managing Taijiquan’s “park culture space” included the democratic operation mechanism of communication and consultation, the cooperative operation mechanism of mutual benefit, and the incentive operation mechanism of balancing interests. Taijiquan’s “park culture space” was the reproduction of public space that was not only bonded with Taijiquan practice but was also reconstructed and expanded by Taijiquan practitioners. Furthermore, it involved the return of Taijiquan practitioners’ historical memory and collective life experience alongside the construction of Taijiquan practitioners’ group identity and the development of self-organization.
Spatiotemporal Change and the Natural–Human Driving Processes of a Megacity’s Coastal Blue Carbon Storage
Authors: Cai Wenbo , Zhu Qing , Chen Meitian , Cai Yongli
Published: August 2021

Coastal blue carbon storage (CBCS) plays a key role in addressing global climate change and realizing regional carbon neutrality. Although blue carbon has been studied for some years, there is little understanding of the influence of a megacity’s complex natural and human-driven processes on CBCS. Taking the Shanghai coastal area as an example, this study investigated the spatiotemporal change in CBCS using the InVEST (Integrated Valuation of Ecosystem Services and Tradeoffs) model during 1990–2015, and analyzed the response of the CBCS to a megacity’s complex natural- and human-driven processes through a land use/land cover transition matrix and hierarchical clustering. The results were as follows: (1) Thirty-three driving processes were identified in the study area, including four natural processes (e.g., accretion, succession, erosion, etc.), two human processes (reclamation and restoration) and twenty-seven natural–human coupled processes; they were further combined into single and multiple processes with positive and negative influences on the CBCS into four types (Mono+, Mono−, Multiple+ and Multiple− driving processes). (2) Shanghai’s CBCS increased from 1659.44 × 104 Mg to 1789.78 ×104 Mg, though the amount of Shanghai’s coastal carbon sequestration showed a decreasing trend in three periods: 51.28 × 104 Mg in 1990–2000, 42.90 × 104 Mg in 2000–2009 and 36.15 × 104 Mg in 2009–2015, respectively. (3) There were three kinds of spatiotemporal patterns in the CBCS of this study area: high adjacent to the territorial land, low adjacent to the offshore waters in 1990; high in the central part, low in the peripheral areas in 2009 and 2015; and a mixed pattern in 2000. These patterns resulted from the different driving processes present in the different years. This study could serve as a blueprint for restoring and maintaining the CBCS of a megacity, to help mitigate the conflicts between socioeconomic development and the conservation of the CBCS, especially in the Shanghai coastal area.
Refinement Proposals for Geodiversity Assessment—A Case Study in the Bakony–Balaton UNESCO Global Geopark, Hungary
Authors: Pál Márton , Albert Gáspár
Published: August 2021

Geodiversity is the variety of natural elements that are excluded from biodiversity, such as: geological, geomorphological, and soil features including their properties, systems, and relationships. Geodiversity assessment measures these features, emphasising the characteristics and physical fragility of the examined areas. In this study, a quantitative methodology has been applied in Bakony–Balaton UGGp, Hungary. The Geopark’s area was divided into 2 × 2 km cells in which geodiversity indices were calculated using various data: maps, spatial databases, and elevation models. However, data sources differ significantly in each country: thematic information may not be entirely public or does not have the appropriate scale and complexity. We proposed to use universal data—geomorphons and a watercourse network—derived from Digital Elevation Models (DEMs) to calculate geomorphological diversity. Making a balance between the base materials was also an aim of this research. As sources with different data densities are used, some abiotic elements may be overrepresented, while others seem to have less significance. The normalisation of thematic layers solves this problem: it gives a proportion to each sub-element and creates a balanced index. By applying worldwide accessible digital base data and statistical standardization methods, abiotic nature quantification may open new perspectives in geoconservation.
The Drought-Mediated Soybean GmNAC085 Functions as a Positive Regulator of Plant Response to Salinity
Authors: Hoang Xuan Lan Thi , Chuong Nguyen Nguyen , Hoa Tran Thi Khanh , Doan Hieu , Van Pham Hoang Phuong , Trang Le Dang Minh , Huyen Pham Ngoc Thai , Le Dung Tien , Tran Lam-Son Phan , Thao Nguyen Phuong
Published: August 2021

Abiotic stress factors, such as drought and salinity, are known to negatively affect plant growth and development. To cope with these adverse conditions, plants have utilized certain defense mechanisms involved in various aspects, including morphological, biochemical and molecular alterations. Particularly, a great deal of evidence for the biological importance of the plant-specific NAM, ATAF1/2, CUC2 (NAC) transcription factors (TFs) in plant adaptation to abiotic stress conditions has been reported. A previous in planta study conducted by our research group demonstrated that soybean (Glycine max) GmNAC085 mediated drought resistance in transgenic Arabidopsis plants. In this study, further characterization of GmNAC085 function in association with salt stress was performed. The findings revealed that under this condition, transgenic soybean plants overexpressing GmNAC085 displayed better germination rates than wild-type plants. In addition, biochemical and transcriptional analyses showed that the transgenic plants acquired a better defense system against salinity-induced oxidative stress, with higher activities of antioxidant enzymes responsible for scavenging hydrogen peroxide or superoxide radicals. Higher transcript levels of several key stress-responsive genes involved in the proline biosynthetic pathway, sodium ion transporter and accumulation of dehydrins were also observed, indicating better osmoprotection and more efficient ion regulation capacity in the transgenic lines. Taken together, these findings and our previous report indicate that GmNAC085 may play a role as a positive regulator in plant adaptation to drought and salinity conditions.
Putative Role of Vitamin D for COVID-19 Vaccination
Authors: Chiu Sheng-Kang , Tsai Kuo-Wang , Wu Chia-Chao , Zheng Cai-Mei , Yang Chung-Hsiang , Hu Wan-Chung , Hou Yi-Chou , Lu Kuo-Cheng , Chao You-Chen
Published: August 2021

Severe acute respiratory syndrome coronavirus 2 is a new, highly pathogenic virus that has recently elicited a global pandemic called the 2019 coronavirus disease (COVID-19). COVID-19 is characterized by significant immune dysfunction, which is caused by strong but unregulated innate immunity with depressed adaptive immunity. Reduced and delayed responses to interferons (IFN-I/IFN-III) can increase the synthesis of proinflammatory cytokines and extensive immune cell infiltration into the airways, leading to pulmonary disease. The development of effective treatments for severe COVID-19 patients relies on our knowledge of the pathophysiological components of this imbalanced innate immune response. Strategies to address innate response factors will be essential. Significant efforts are currently underway to develop vaccines against SARS-CoV-2. COVID-19 vaccines, such as inactivated DNA, mRNA, and protein subunit vaccines, have already been applied in clinical use. Various vaccines display different levels of effectiveness, and it is important to continue to optimize and update their composition in order to increase their effectiveness. However, due to the continuous emergence of variant viruses, improving the immunity of the general public may also increase the effectiveness of the vaccines. Many observational studies have demonstrated that serum levels of vitamin D are inversely correlated with the incidence or severity of COVID-19. Extensive evidence has shown that vitamin D supplementation could be vital in mitigating the progression of COVID-19 to reduce its severity. Vitamin D defends against SARS-CoV-2 through a complex mechanism through interactions between the modulation of innate and adaptive immune reactions, ACE2 expression, and inhibition of the renin-angiotensin system (RAS). However, it remains unclear whether Vit-D also plays an important role in the effectiveness of different COVID-19 vaccines. Based on analysis of the molecular mechanism involved, we speculated that vit-D, via various immune signaling pathways, plays a complementary role in the development of vaccine efficacy.
Functional Dental Pulp Regeneration: Basic Research and Clinical Translation
Authors: Xie Zhuo , Shen Zongshan , Zhan Peimeng , Yang Jiayu , Huang Qiting , Huang Shuheng , Chen Lingling , Lin Zhengmei
Published: August 2021

Pulpal and periapical diseases account for a large proportion of dental visits, the current treatments for which are root canal therapy (RCT) and pulp revascularisation. Despite the clinical signs of full recovery and histological reconstruction, true regeneration of pulp tissues is still far from being achieved. The goal of regenerative endodontics is to promote normal pulp function recovery in inflamed or necrotic teeth that would result in true regeneration of the pulpodentinal complex. Recently, rapid progress has been made related to tissue engineering-mediated pulp regeneration, which combines stem cells, biomaterials, and growth factors. Since the successful isolation and characterisation of dental pulp stem cells (DPSCs) and other applicable dental mesenchymal stem cells, basic research and preclinical exploration of stem cell-mediated functional pulp regeneration via cell transplantation and cell homing have received considerably more attention. Some of this effort has translated into clinical therapeutic applications, bringing a ground-breaking revolution and a new perspective to the endodontic field. In this article, we retrospectively examined the current treatment status and clinical goals of pulpal and periapical diseases and scrutinized biological studies of functional pulp regeneration with a focus on DPSCs, biomaterials, and growth factors. Then, we reviewed preclinical experiments based on various animal models and research strategies. Finally, we summarised the current challenges encountered in preclinical or clinical regenerative applications and suggested promising solutions to address these challenges to guide tissue engineering-mediated clinical translation in the future.
Angioedema and Fatty Acids
Authors: Wada Akane , Sawada Yu , Sugino Hitomi , Nakamura Motonobu
Published: August 2021

Angioedema is a life-threatening emergency event that is associated with bradykinin and histamine-mediated cascades. Although bradykinin-mediated angioedema currently has specific therapeutic options, angioedema is sometimes intractable with current treatments, especially histamine-mediated angioedema, suggesting that some other mediators might contribute to the development of angioedema. Fatty acids are an essential fuel and cell component, and act as a mediator in physiological and pathological human diseases. Recent updates of studies revealed that these fatty acids are involved in vascular permeability and vasodilation, in addition to bradykinin and histamine-mediated reactions. This review summarizes each fatty acid’s function and the specific receptor signaling responses in blood vessels, and focuses on the possible pathogenetic role of fatty acids in angioedema.
Photopharmacological Applications for Cherenkov Radiation Generated by Clinically Used Radionuclides
Authors: Krebs Melanie , Döbber Alexander , Rodat Theo , Lützen Ulf , Zhao Yi , Zuhayra Maaz , Peifer Christian
Published: August 2021

Translational photopharmacological applications are limited through irradiation by light showing wavelengths within the bio-optical window. To achieve sufficient tissue penetration, using wavelengths >500 nm is mandatory. Nevertheless, the majority of photopharmacological compounds respond to irradiation with more energetic UV light, which shows only a minor depth of tissue penetration in the µm range. Thus, we became interested in UV light containing Cherenkov radiation (CR) induced as a by-product by clinically employed radionuclides labeling specific tissues. Therefore, CR may be applicable in novel photopharmacological approaches. To provide evidence for the hypothesis, we verified the clinically established radionuclides 68Ga and 90Y but not 18F in clinically used activities to be capable of generating CR in aqueous solutions. We then investigated whether the generated CR was able to photoactivate the caged kinase inhibitor cagedAZD5438 as a photoresponsive model system. Herein, 21% uncaging of the model system cagedAZD5438 occurred by incubation with 90Y, along with a non-specific compound decomposition for 68Ga and partly for 90Y. The findings suggest that the combination of a clinically employed radionuclide with an optimized photoresponsive agent could be beneficial for highly focused photopharmacological therapies.
Zebrafish Model as a Screen to Prevent Cyst Inflation in Autosomal Dominant Polycystic Kidney Disease
Authors: Oliveira Inês , Jacinto Raquel , Pestana Sara , Nolasco Fernando , Calado Joaquim , Lopes Susana Santos , Roxo-Rosa Mónica
Published: August 2021

In autosomal dominant polycystic kidney disease (ADPKD), kidney cyst growth requires the recruitment of CFTR (cystic fibrosis transmembrane conductance regulator), the chloride channel that is defective in cystic fibrosis. We have been studying cyst inflation using the zebrafish Kupffer’s vesicle (KV) as model system because we previously demonstrated that knocking down polycystin 2 (PC2) induced a CFTR-mediated enlargement of the organ. We have now quantified the PC2 knockdown by showing that it causes a 73% reduction in the number of KV cilia expressing PC2. According to the literature, this is an essential event in kidney cystogenesis in ADPKD mice. Additionally, we demonstrated that the PC2 knockdown leads to a significant accumulation of CFTR-GFP at the apical region of the KV cells. Furthermore, we determined that KV enlargement is rescued by the injection of Xenopus pkd2 mRNA and by 100 µM tolvaptan treatment, the unique and approved pharmacologic approach for ADPKD management. We expected vasopressin V2 receptor antagonist to lower the cAMP levels of KV-lining cells and, thus, to inactivate CFTR. These findings further support the use of the KV as an in vivo model for screening compounds that may prevent cyst enlargement in this ciliopathy, through CFTR inhibition.
A Gain-of-Function Mutant of IAA7 Inhibits Stem Elongation by Transcriptional Repression of EXPA5 Genes in Brassica napus
Authors: Wei Tao , Zhang Li , Zhu Ruijia , Jiang Xuefei , Yue Chu , Su Ying , Ren Hongpei , Wang Maolin
Published: August 2021

Plant height is one of the most important agronomic traits of rapeseeds. In this study, we characterized a dwarf Brassica napus mutant, named ndf-2, obtained from fast neutrons and DES mutagenesis. Based on BSA-Seq and genetic properties, we identified causal mutations with a time-saving approach. The ndf-2 mutation was identified on chromosome A03 and can result in an amino acid substitution in the conserved degron motif (GWPPV to EWPPV) of the Auxin/indole-3-acetic acid protein 7 (BnaA03.IAA7) encoded by the causative gene. Aux/IAA protein is one of the core components of the auxin signaling pathway, which regulates many growth and development processes. However, the molecular mechanism of auxin signal regulating plant height is still not well understood. In the following work, we identified that BnaARF6 and BnaARF8 as interactors of BnaA03.IAA7 and BnaEXPA5 as a target of BnaARF6 and BnaARF8. The three genes BnaA03.IAA7, BnaARF6/8 and BnaEXPA5 were highly expressed in stem, suggesting that these genes were involved in stem development. The overexpression of BnaEXPA5 results in larger rosettes leaves and longer inflorescence stems in Arabidopsis thaliana. Our results indicate that BnaA03.IAA7- and BnaARF6/8-dependent auxin signal control stem elongation and plant height by regulating the transcription of BnaEXPA5 gene, which is one of the targets of this signal.
Involvement of Virus-Induced Interferon Production in IgG Autoantibody-Mediated Anemia
Authors: Legrain Sarah , Su Dan , Gaignage Mélanie , Breukel Cor , Claassens Jill , Brouwers Conny , Linssen Margot M. , Izui Shozo , Verbeek J. Sjef , Coutelier Jean-Paul
Published: August 2021

Infection with viruses, such as the lactate dehydrogenase-elevating virus (LDV), is known to trigger the onset of autoimmune anemia through the enhancement of the phagocytosis of autoantibody-opsonized erythrocytes by activated macrophages. Type I interferon receptor-deficient mice show enhanced anemia, which suggests a protective effect of these cytokines, partly through the control of type II interferon production. The development of anemia requires the expression of Fcγ receptors (FcγR) I, III, and IV. Whereas LDV infection decreases FcγR III expression, it enhances FcγR I and IV expression in wild-type animals. The LDV-associated increase in the expression of FcγR I and IV is largely reduced in type I interferon receptor-deficient mice, through both type II interferon-dependent and -independent mechanisms. Thus, the regulation of the expression of FcγR I and IV, but not III, by interferons may partly explain the exacerbating effect of LDV infection on anemia that results from the enhanced phagocytosis of IgG autoantibody-opsonized erythrocytes.
Effects on Metabolism in Astrocytes Caused by cGAMP, Which Imitates the Initial Stage of Brain Metastasis
Authors: Okawa Toya , Hara Kurumi , Goto Momoko , Kikuchi Moe , Kogane Masataka , Hatakeyama Hiroto , Tanaka Hiroki , Shirane Daiki , Akita Hidetaka , Hisaka Akihiro , Sato Hiromi
Published: August 2021

The second messenger 2′3′-cyclic-GMP-AMP (cGAMP) is thought to be transmitted from brain carcinomas to astrocytes via gap junctions, which functions to promote metastasis in the brain parenchyma. In the current study, we established a method to introduce cGAMP into astrocytes, which simulates the state of astrocytes that have been invaded by cGAMP around tumors. Astrocytes incorporating cGAMP were analyzed by metabolomics, which demonstrated that cGAMP increased glutamate production and astrocyte secretion. The same trend was observed for γ-aminobutyric acid (GABA). Conversely, glutamine production and secretion were decreased by cGAMP treatment. Due to the fundamental role of astrocytes in regulation of the glutamine–glutamate cycle, such metabolic changes may represent a potential mechanism and therapeutic target for alteration of the central nervous system (CNS) environment and the malignant transformation of brain carcinomas.
Activation of Neuronal Nicotinic Receptors Inhibits Acetylcholine Release in the Neuromuscular Junction by Increasing Ca2+ Flux through Cav1 Channels
Authors: Zhilyakov Nikita , Arkhipov Arsenii , Malomouzh Artem , Samigullin Dmitry
Published: August 2021

Cholinergic neurotransmission is a key signal pathway in the peripheral nervous system and in several branches of the central nervous system. Despite the fact that it has been studied extensively for a long period of time, some aspects of its regulation still have not yet been established. One is the relationship between the nicotine-induced autoregulation of acetylcholine (ACh) release with changes in the concentration of presynaptic calcium levels. The mouse neuromuscular junction of m. Levator Auris Longus was chosen as the model of the cholinergic synapse. ACh release was assessed by electrophysiological methods. Changes in calcium transients were recorded using a calcium-sensitive dye. Nicotine hydrogen tartrate salt application (10 μM) decreased the amount of evoked ACh release, while the calcium transient increased in the motor nerve terminal. Both of these effects of nicotine were abolished by the neuronal ACh receptor antagonist dihydro-beta-erythroidine and Cav1 blockers, verapamil, and nitrendipine. These data allow us to suggest that neuronal nicotinic ACh receptor activation decreases the number of ACh quanta released by boosting calcium influx through Cav1 channels.
Targeting Lysosomes to Reverse Hydroquinone-Induced Autophagy Defects and Oxidative Damage in Human Retinal Pigment Epithelial Cells
Authors: Abokyi Samuel , Shan Sze-Wan , Lam Christie Hang-I , Catral Kirk Patrick , Pan Feng , Chan Henry Ho-Lung , To Chi-Ho , Tse Dennis Yan-Yin
Published: August 2021

In age-related macular degeneration (AMD), hydroquinone (HQ)-induced oxidative damage in retinal pigment epithelium (RPE) is believed to be an early event contributing to dysregulation of inflammatory cytokines and vascular endothelial growth factor (VEGF) homeostasis. However, the roles of antioxidant mechanisms, such as autophagy and the ubiquitin-proteasome system, in modulating HQ-induced oxidative damage in RPE is not well-understood. This study utilized an in-vitro AMD model involving the incubation of human RPE cells (ARPE-19) with HQ. In comparison to hydrogen peroxide (H2O2), HQ induced fewer reactive oxygen species (ROS) but more oxidative damage as characterized by protein carbonyl levels, mitochondrial dysfunction, and the loss of cell viability. HQ blocked the autophagy flux and increased proteasome activity, whereas H2O2 did the opposite. Moreover, the lysosomal membrane-stabilizing protein LAMP2 and cathepsin D levels declined with HQ exposure, suggesting loss of lysosomal membrane integrity and function. Accordingly, HQ induced lysosomal alkalization, thereby compromising the acidic pH needed for optimal lysosomal degradation. Pretreatment with MG132, a proteasome inhibitor and lysosomal stabilizer, upregulated LAMP2 and autophagy and prevented HQ-induced oxidative damage in wildtype RPE cells but not cells transfected with shRNA against ATG5. This study demonstrated that lysosomal dysfunction underlies autophagy defects and oxidative damage induced by HQ in human RPE cells and supports lysosomal stabilization with the proteasome inhibitor MG132 as a potential remedy for oxidative damage in RPE and AMD.
Redox Signaling and Sarcopenia: Searching for the Primary Suspect
Authors: Foreman Nicholas A. , Hesse Anton S. , Ji Li Li
Published: August 2021

Sarcopenia, the age-related decline in muscle mass and function, derives from multiple etiological mechanisms. Accumulative research suggests that reactive oxygen species (ROS) generation plays a critical role in the development of this pathophysiological disorder. In this communication, we review the various signaling pathways that control muscle metabolic and functional integrity such as protein turnover, cell death and regeneration, inflammation, organismic damage, and metabolic functions. Although no single pathway can be identified as the most crucial factor that causes sarcopenia, age-associated dysregulation of redox signaling appears to underlie many deteriorations at physiological, subcellular, and molecular levels. Furthermore, discord of mitochondrial homeostasis with aging affects most observed problems and requires our attention. The search for the primary suspect of the fundamental mechanism for sarcopenia will likely take more intense research for the secret of this health hazard to the elderly to be unlocked.
Evaluation of Epigenetic and Radiomodifying Effects during Radiotherapy Treatments in Zebrafish
Authors: Pucci Gaia , Forte Giusi Irma , Cavalieri Vincenzo
Published: August 2021

Radiotherapy is still a long way from personalizing cancer treatment plans, and its effectiveness depends on the radiosensitivity of tumor cells. Indeed, therapies that are efficient and successful for some patients may be relatively ineffective for others. Based on this, radiobiological research is focusing on the ability of some reagents to make cancer cells more responsive to ionizing radiation, as well as to protect the surrounding healthy tissues from possible side effects. In this scenario, zebrafish emerged as an effective model system to test for radiation modifiers that can potentially be used for radiotherapeutic purposes in humans. The adoption of this experimental organism is fully justified and supported by the high similarity between fish and humans in both their genome sequences and the effects provoked in them by ionizing radiation. This review aims to provide the literature state of the art of zebrafish in vivo model for radiobiological studies, particularly focusing on the epigenetic and radiomodifying effects produced during fish embryos’ and larvae’s exposure to radiotherapy treatments.
Iterated Virtual Screening-Assisted Antiviral and Enzyme Inhibition Assays Reveal the Discovery of Novel Promising Anti-SARS-CoV-2 with Dual Activity
Authors: Hamdy Rania , Fayed Bahgat , Mostafa Ahmed , Shama Noura M. Abo , Mahmoud Sara Hussein , Mehta Chetan Hasmukh , Nayak Yogendra , M. Soliman Sameh S.
Published: August 2021

Unfortunately, COVID-19 is still a threat to humankind and has a dramatic impact on human health, social life, the world economy, and food security. With the limited number of suggested therapies under clinical trials, the discovery of novel therapeutic agents is essential. Here, a previously identified anti-SARS-CoV-2 compound named Compound 13 (1,2,5-Oxadiazole-3-carboximidic acid, 4,4′-(methylenediimino) bis,bis[[(2-hydroxyphenyl)methylene]hydrazide) was subjected to an iterated virtual screening against SARS-CoV-2 Mpro using a combination of Ligand Designer and PathFinder. PathFinder, a computational reaction enumeration tool, was used for the rapid generation of enumerated structures via default reaction library. Ligand designer was employed for the computerized lead optimization and selection of the best structural modification that resulted in a favorable ligand–protein complex. The obtained compounds that showed the best binding to Mpro were re-screened against TMPRSS2, leading to the identification of 20 shared compounds. The compounds were further visually inspected, which resulted in the identification of five shared compounds M1–5 with dual binding affinity. In vitro evaluation and enzyme inhibition assay indicated that M3, an analogue of Compound 13 afforded by replacing the phenolic moiety with pyridinyl, possesses an improved antiviral activity and safety. M3 displayed in vitro antiviral activity with IC50 0.016 µM and Mpro inhibition activity with IC50 0.013 µM, 7-fold more potent than the parent Compound 13 and potent than the antivirals drugs that are currently under clinical trials. Moreover, M3 showed potent activity against human TMPRSS2 and furin enzymes with IC50 0.05, and 0.08 µM, respectively. Molecular docking, WaterMap analysis, molecular dynamics simulation, and R-group analysis confirmed the superiority of the binding fit to M3 with the target enzymes. WaterMap analysis calculated the thermodynamic properties of the hydration site in the binding pocket that significantly affects the biological activity. Loading M3 on zinc oxide nanoparticles (ZnO NPs) increased the antiviral activity of the compound 1.5-fold, while maintaining a higher safety profile. In conclusion, lead optimized discovery following an iterated virtual screening in association with molecular docking and biological evaluation revealed a novel compound named M3 with promising dual activity against SARS-CoV-2. The compound deserves further investigation for potential clinical-based studies.
Reconstruction of the Diaminopimelic Acid Pathway to Promote L-lysine Production in Corynebacterium glutamicum
Authors: Liu Ning , Zhang Ting-Ting , Rao Zhi-Ming , Zhang Wei-Guo , Xu Jian-Zhong
Published: August 2021

The dehydrogenase pathway and the succinylase pathway are involved in the synthesis of L-lysine in Corynebacterium glutamicum. Despite the low contribution rate to L-lysine production, the dehydrogenase pathway is favorable for its simple steps and potential to increase the production of L-lysine. The effect of ammonium (NH4+) concentration on L-lysine biosynthesis was investigated, and the results indicated that the biosynthesis of L-lysine can be promoted in a high NH4+ environment. In order to reduce the requirement of NH4+, the nitrogen source regulatory protein AmtR was knocked out, resulting in an 8.5% increase in L-lysine production (i.e., 52.3 ± 4.31 g/L). Subsequently, the dehydrogenase pathway was upregulated by blocking or weakening the tetrahydrodipicolinate succinylase (DapD)-coding gene dapD and overexpressing the ddh gene to further enhance L-lysine biosynthesis. The final strain XQ-5-W4 could produce 189 ± 8.7 g/L L-lysine with the maximum specific rate (qLys,max.) of 0.35 ± 0.05 g/(g·h) in a 5-L jar fermenter. The L-lysine titer and qLys,max achieved in this study is about 25.2% and 59.1% higher than that of the original strain without enhancement of dehydrogenase pathway, respectively. The results indicated that the dehydrogenase pathway could serve as a breakthrough point to reconstruct the diaminopimelic acid (DAP) pathway and promote L-lysine production.
Contrasting Local and Macroscopic Effects of Collagen Hydroxylation
Authors: Varma Sameer , Orgel Joseph P. R. O. , Schieber Jay D.
Published: August 2021

Collagen is heavily hydroxylated. Experiments show that proline hydroxylation is important to triple helix (monomer) stability, fibril assembly, and interaction of fibrils with other molecules. Nevertheless, experiments also show that even without hydroxylation, type I collagen does assemble into its native D-banded fibrillar structure. This raises two questions. Firstly, even though hydroxylation removal marginally affects macroscopic structure, how does such an extensive chemical change, which is expected to substantially reduce hydrogen bonding capacity, affect local structure? Secondly, how does such a chemical perturbation, which is expected to substantially decrease electrostatic attraction between monomers, affect collagen’s mechanical properties? To address these issues, we conduct a benchmarked molecular dynamics study of rat type I fibrils in the presence and absence of hydroxylation. Our simulations reproduce the experimental observation that hydroxylation removal has a minimal effect on collagen’s D-band length. We also find that the gap-overlap ratio, monomer width and monomer length are minimally affected. Surprisingly, we find that de-hydroxylation also has a minor effect on the fibril’s Young’s modulus, and elastic stress build up is also accompanied by tightening of triple-helix windings. In terms of local structure, de-hydroxylation does result in a substantial drop (23%) in inter-monomer hydrogen bonding. However, at the same time, the local structures and inter-monomer hydrogen bonding networks of non-hydroxylated amino acids are also affected. It seems that it is this intrinsic plasticity in inter-monomer interactions that preclude fibrils from undergoing any large changes in macroscopic properties. Nevertheless, changes in local structure can be expected to directly impact collagen’s interaction with extra-cellular matrix proteins. In general, this study highlights a key challenge in tissue engineering and medicine related to mapping collagen chemistry to macroscopic properties but suggests a path forward to address it using molecular dynamics simulations.
Increased Potential of Bone Formation with the Intravenous Injection of a Parathyroid Hormone-Related Protein Minicircle DNA Vector
Authors: Kim Jang-Woon , Park Narae , Kang Jaewoo , Kim Yena , Jung Hyerin , Rim Yeri Alice , Ju Ji Hyeon
Published: August 2021

Osteoporosis is commonly treated via the long-term usage of anti-osteoporotic agents; however, poor drug compliance and undesirable side effects limit their treatment efficacy. The parathyroid hormone-related protein (PTHrP) is essential for normal bone formation and remodeling; thus, may be used as an anti-osteoporotic agent. Here, we developed a platform for the delivery of a single peptide composed of two regions of the PTHrP protein (1–34 and 107–139); mcPTHrP 1–34+107–139 using a minicircle vector. We also transfected mcPTHrP 1–34+107–139 into human mesenchymal stem cells (MSCs) and generated Thru 1–34+107–139-producing engineered MSCs (eMSCs) as an alternative delivery system. Osteoporosis was induced in 12-week-old C57BL/6 female mice via ovariectomy. The ovariectomized (OVX) mice were then treated with the two systems; (1) mcPTHrP 1–34+107–139 was intravenously administered three times (once per week); (2) eMSCs were intraperitoneally administered twice (on weeks four and six). Compared with the control OVX mice, the mcPTHrP 1–34+107–139-treated group showed better trabecular bone structure quality, increased bone formation, and decreased bone resorption. Similar results were observed in the eMSCs-treated OVX mice. Altogether, these results provide experimental evidence to support the potential of delivering PTHrP 1–34+107–139 using the minicircle technology for the treatment of osteoporosis.
Simultaneous Monitoring of Monoclonal Antibody Variants by Strong Cation-Exchange Chromatography Hyphenated to Mass Spectrometry to Assess Quality Attributes of Rituximab-Based Biotherapeutics
Authors: Di Marco Fiammetta , Berger Thomas , Esser-Skala Wolfgang , Rapp Erdmann , Regl Christof , Huber Christian G.
Published: August 2021

Different manufacturing processes and storage conditions of biotherapeutics can lead to a significant variability in drug products arising from chemical and enzymatic post-translational modifications (PTMs), resulting in the co-existence of a plethora of proteoforms with different physicochemical properties. To unravel the heterogeneity of these proteoforms, novel approaches employing strong cation-exchange (SCX) high-performance liquid chromatography (HPLC) hyphenated to mass spectrometry (MS) using a pH gradient of volatile salts have been developed in recent years. Here, we apply an established SCX-HPLC-MS method to characterize and compare two rituximab-based biotherapeutics, the originator MabThera® and its Indian copy product Reditux™. The study assessed molecular differences between the two drug products in terms of C-terminal lysine variants, glycosylation patterns, and other basic and acidic variants. Overall, MabThera® and Reditux™ displayed differences at the molecular level. MabThera® showed a higher degree of galactosylated and sialylated glycoforms, while Reditux™ showed increased levels of oligomannose and afucosylated glycoforms. Moreover, the two drug products showed differences in terms of basic variants such as C-terminal lysine and N-terminal truncation, present in Reditux™ but not in MabThera®. This study demonstrates the capability of this fast SCX-HPLC-MS approach to compare different drug products and simultaneously assess some of their quality attributes.