bluuebunny/crossref_metadata_2025 huggingface dataset:
Genome-Wide Identification of the 1-Aminocyclopropane-1-carboxylic Acid Synthase (ACS) Genes and Their Possible Role in Sand Pear (Pyrus pyrifolia) Fruit Ripening
Authors: Zhang Jing-Guo , Du Wei , Fan Jing , Yang Xiao-Ping , Chen Qi-Liang , Liu Ying , Hu Hong-Ju , Luo Zheng-Rong
Published: October 2021

Ethylene production is negatively associated with storage life in sand pear (Pyrus pyrifolia Nakai), particularly at the time of fruit harvest. 1-Aminocyclopropane-1-carboxylic acid synthase (ACS) is the rate-limiting enzyme in ethylene biosynthesis and is considered to be important for fruit storage life. However, the candidate ACS genes and their roles in sand pear remain unclear. The present study identified 13 ACS genes from the sand pear genome. Phylogenetic analysis categorized these ACS genes into four subgroups (type I, type II, type III and putative AAT), and indicated a close relationship between sand pear and Chinese white pear (P. bretschneideri). According to the RNA-seq data and qRT-PCR analysis, PpyACS1, PpyACS2, PpyACS3, PpyACS8, PpyACS9, PpyACS12 and PpyACS13 were differently expressed in climacteric and non-climacteric-type pear fruits, ‘Ninomiyahakuri’ and ‘Eli No.2’, respectively, during fruit ripening. In addition, the expressions of PpyACS2, PpyACS8, PpyACS12 and PpyACS13 were found to be associated with system 1 of ethylene production, while PpyACS1, PpyACS3, and PpyACS9 were found to be associated with system 2, indicating that these ACS genes have different roles in ethylene biosynthesis during fruit development. Overall, our study provides fundamental knowledge on the characteristics of the ACS gene family in sand pear, in addition to their possible roles in fruit ripening.
Variability of Polyphenolic Compounds and Biological Activities among Perilla frutescens var. crispa Genotypes
Authors: Ju Hyun Ju , Kim Kyeoung Cheol , Kim Heekyu , Kim Ju-Sung , Hyun Tae Kyung
Published: October 2021

Perilla frutescens var. crispa (Pfc) of the family Lamiaceae is used as a medicinal plant due to its pharmacological properties. Although Pfc is an important resource for the medical nutrition industry, the variability in phytonutrients and biological activities among genotypes of Pfc is not well understood. The effects of genotype on the phytochemical composition, antioxidant activities, antimelanogenic principles, and anti-inflammatory effects of Pfc were determined using eight Pfc genotypes. Using HPLC analysis, we identified 30 polyphenolic compounds from Pfc, although variation was observed in the polyphenolic composition of Pfc genotypes. Pfc 5 exhibited antimelanogenic activity in B16F10 melanoma cells via inhibition of tyrosinase activity. In addition, Pfc 2 strongly inhibited lipopolysaccharide-induced nitric oxide production through translational downregulation of inducible NOS in RAW264 murine macrophages. Taken together, the results of our study reveal the significant impacts of genotype on phytonutrients and biological activities. This finding will assist in the breeding and genetic engineering of Pfc in order to meet future phytonutrition and health challenges.
Concentrations and Risk Assessments of Antibiotics in an Urban–Rural Complex Watershed with Intensive Livestock Farming
Authors: Lee Hyun-Jeoung , Kim Deok-Woo , Kim Chansik , Ryu Hong-Duck , Chung Eu-Gene , Kim Kyunghyun
Published: October 2021

Antibiotics used for the treatment of humans and livestock are released into the environment, whereby they pose a grave threat to biota (including humans) as they can cause the emergence of various strains of resistant bacteria. An improved understanding of antibiotics in the environment is thus vital for appropriate management and mitigation. Herein, surface water and groundwater samples containing antibiotics were analyzed in an urban–rural complex watershed (Cheongmi Stream) comprising intensive livestock farms by collecting samples across different time points and locations. The spatiotemporal trends of the residual antibiotics were analyzed, and ecological and antibiotic resistance-based risk assessments were performed considering their concentrations. The results showed that the concentrations and detection frequencies of the residual antibiotics in the surface water were affected by various factors such as agricultural activities and point sources, and were higher than those found in groundwater; however, frequent detection of antibiotics in groundwater showed that residual antibiotics were influenced by factors such as usage pattern and sewage runoff. Furthermore, few antibiotics posed ecological risks. The risk assessment methods adopted in this study can be applied elsewhere, and the results can be considered in the environmental management of residual antibiotics in the Cheongmi Stream watershed.
Trait Emotional Intelligence and Happiness of Young Adults: The Mediating Role of Perfectionism
Authors: Badri Siti Khadijah Zainal , Kong Min Yao , Wan Mohd Yunus Wan Mohd Azam , Nordin Nor Akmar , Yap Wai Meng
Published: October 2021

Perfectionism or a tendency to aim for an unrealistic standard can impair happiness. However, the potential mechanisms of perfectionism to explain the association between trait emotional intelligence (EI) and happiness are still understudied. This study explores the mediating role of perfectionism in the relationship between trait emotional intelligence (EI) and happiness among young adults. A cross-sectional sample of 259 young adults aged between 18 to 35 years old was recruited. All analyses were conducted using SPSS and AMOS Structural Equation Modeling. High trait EI was linked to low perfectionism and high happiness levels. Furthermore, perfectionism mediated the relationship between trait EI and happiness. Although high trait EI lowered maladaptive perfectionism, the negative impact of maladaptive perfectionism remained and subsequently led to decreasing happiness levels of young adults. This study offers an enhanced understanding of the role of perfectionism in explaining the happiness state of young adults. Moreover, it provides practical implications for using trait EI and managing perfectionism tendency to manage the happiness and wellbeing of the young adult population.
The Impact of Carbon Trade on Outsourcing Remanufacturing
Authors: Xia Xiqiang , Li Mengya , Li Biao , Wang Hao
Published: October 2021

Outsourcing remanufacturing is an important way to achieve resource recycling, green manufacturing and carbon neutrality goals. To analyze the impact of carbon trade on manufacturing/remanufacturing under outsourcing remanufacturing, this article builds a game model between an original equipment manufacturer (OEM) and a remanufacturer under the carbon trade policy. In the outsourcing remanufacturing model, this article compares the impact of the carbon trade policy on the unit retail price, sales volume, revenue, environmental impact, and consumer surplus of new and remanufactured products. The research mainly draws the following conclusions: (1) Carbon trade increases the prices of both new and remanufactured products and the cost of outsourcing. Only when certain conditions are met can increased carbon trade prices increase revenue. (2) The carbon trade policy helps reduce the adverse impact on the environment, but only when the carbon trade price is greater than a certain threshold can it increase consumer surplus. (3) Consumer preferences and carbon emissions of the unit product affect manufacturers’ profits. Increased consumer preference for remanufactured products and reduced carbon emissions of remanufactured products contribute to increased sales and revenues.
Antecedents of Workplace Bullying among Employees in Germany: Five-Year Lagged Effects of Job Demands and Job Resources
Authors: Conway Paul Maurice , Burr Hermann , Rose Uwe , Clausen Thomas , Balducci Cristian
Published: October 2021

Objectives: The aim of the present study was to examine the long-term association of job demands and job resources with self-reported exposure to workplace bullying in a representative sample of employees in Germany. Methods: We analysed a nation-wide representative cohort of employees working in the same workplace with a 5-year follow-up (S-MGA; N = 1637). The study contained self-reported measures of psychosocial working conditions, including work pace, amount of work, influence at work, role clarity and quality of leadership, and workplace bullying, and of organisational factors, including organisational restructuring and layoffs. Results: After controlling for bullying and occupational level at baseline, higher baseline levels of organisational restructuring (OR 1.73; 95% CI 1.10–2.70), work pace (1.30; 95% CI 1.01–1.66), and amount of work (1.55; 95% CI 1.21–1.99), and lower baseline levels of influence at work (0.70; 95% CI 0.55–0.90) and quality of leadership (0.64; 95% CI 0.50–0.82), were associated with an elevated risk of workplace bullying at follow-up. In all, 90% of cases of self-reported workplace bullying could be attributed to these factors. Conclusions: The study suggests that employees reporting higher demands and lower resources, as well as organisational factors such as restructuring, are at a higher long-term risk of being targets of workplace bullying. Interventions aimed at preventing workplace bullying could benefit from a focus on psychosocial working conditions and organisational factors.
Do Blood Lactate Levels Affect the Kinematic Patterns of Jump Shots in Handball?
Authors: Belcic Ivan , Rodić Sasa , Dukarić Vedran , Rupčić Tomislav , Knjaz Damir
Published: October 2021

The aim of this study was to determine whether the dynamic motor stereotype of movement (shooting technique) is violated under conditions of an increased lactate concentration in a player’s blood after a 30–15 intermittent fitness test. The hypotheses was that there would be statistically significant differences in ball speed and shooting accuracy in jump shots on the goal before and after the occurrence of fatigue in the player. The sample of respondents consisted of 10 top-level handball players of the highest competition rank in Croatia. The results showed significant differences before and after the fatigue protocol in the run-up speed (F = 5.66; p = 0.02), in the maximum speed of the forearm (F = 5.85; p = 0.02) and the hand (F = 4.01; p = 0.04), in the speed in the shoulder joint (F = 5.39; p = 0.02) and wrist joint (F = 4.06; p = 0.04), and in the ball shooting speed (F = 5.42; p = 0.02). The accuracy of the shot was, on average, lower (36.20 vs. 33.17 cm) but not significantly so. High blood lactate levels affect changes in certain kinematic parameters during the performance of a jump shot in handball. Consequently, this reduces the speed of the shot, which can affect situational performance as one of the two significant parameters of scoring success.
Urban-Rural Differences in Physical Fitness and Out-of-School Physical Activity for Primary School Students: A County-Level Comparison in Western China
Authors: Tian Yunxi , Liu Lingfang , Wang Xuhui , Zhang Xue , Zhai Yang , Wang Kai , Liu Jianjun
Published: October 2021

Rapid urbanization of China has brought lifestyle changes resulting in a continuous decline in children’s physical fitness (PF) and out-of-school physical activity (PA). To date, studies have been focused on correlates of PF and out-of-school PA, and patterns and trends based on geographic diversity and urban-rural contrasts. Western China, with a large rural population, has substantial urban-rural differences, but little work has been done to compare its children’s physical fitness (PF) and out-of-school physical activity (PA) at a county level. A total of 715 primary school students (grades 3–6) were surveyed from one urban school (n = 438) and four rural schools (n = 277) in a county-level administrative unit, Yangling District, Shaanxi, in western China. Physical fitness index (PFI) was measured and calculated based on the revised Chinese Student Physical Fitness Standards. Out-of-school PA and other variables of demographics, behavior and perception were collected using questionnaires. Statistical analyses explored urban-rural differences and correlates of PFI and out-of-school PA. We found that the PFI (72.86 vs. 79.67) and weekly moderate-to-vigorous physical activity (MVPA) duration (167.57 vs. 220.08) of urban students were significantly lower than those of rural students. Weekly MVPA duration had the largest positive impact on PFI. Perceived availability of PA spaces was positively associated with both the urban and rural students’ PF and PA, while screen time was negatively associated with PF and PA, especially for rural students. Facilitators of PA frequency include the perceived availability of PA time and parental educational level. Parents’ PA habits had a positive impact on urban students’ PA. No association between active school commuting and PF or PA was found. Our findings revealed that PF and out-of-school PA of urban students were clearly lower than among rural students. The health of rural children at the county level in western China should be paid much more attention during the process of rapid urbanization.
Educational Programs for the Promotion of Health at School: A Systematic Review
Authors: Pérez-Jorge David , González-Luis María Alejandra , Rodríguez-Jiménez María del Carmen , Ariño-Mateo Eva
Published: October 2021

Context: Health promotion programs generate healthy changes in the educational community. However, not all of them meet the expected objectives due to multiple factors that affect their development, such as the teachers overload work, the lack of specific training, the lack of time to carry out health promotion activities, the lack of flexibility of the programs, and their non-inclusion in the training programs of the centers. Objective: To know the scope of the strategies and programs that promote healthy habits among students in compulsory educational stages. Data sources: a systematic review of articles in English, using the Web of Science (WOS), Medline, and PsycINFO databases.
Association of COMT Polymorphisms with Multiple Physical Activity-Related Injuries among University Students in China
Authors: Chen Shangmin , Cai Weicong , Duan Shiwei , Gao Lijie , Yang Wenda , Gao Yang , Jia Cunxian , Zhang Hongjuan , Li Liping
Published: October 2021

The catechol-O-methyltransferase (COMT) is a candidate gene to provide promising evidence of psychiatric disorders, but there is a knowledge gap between the genetic factor and multiple physical activity-related injuries (PARIs). The aim of this study was to explore the contribution of COMT to the risk of PARIs among university students in the Chinese Han population. We can further search for the intrinsic risk factors for the occurrence of multiple physical activity injuries and provide a scientific basis for early screening and precise intervention for the high-risk group of college students with multiple PARIs. A 1:1 matched case-control study of 61 PARIs cases and 61 healthy controls were carried out. DNA samples of the participants were isolated from saliva and genotyped on eight SNPs of the COMT gene (rs9265, rs4680, rs6269, rs4818, rs4633, rs165655, rs165656, and rs165722) using the MALDI-TOF MS method. We found that rs6269 and rs4818 were significantly associated with PARIs, and rs6269-GG and rs4818-GG contributed to the reduced risk of PARIs. Further haplotype analysis showed a four-marker C-G-C-G haplotype (rs165722-rs6269-rs4633-rs4818) acted with a protective role in the development of PARIs (p = 0.037; OR: 0.474, 95% CI: 0.269 to 0.834). However, the interactions between club membership and rs6269 or rs4818 would significantly increase the risk of PARIs (both p < 0.001, OR: 5.121 and 4.977, respectively). This is the first study to find the contribution of COMT to PARIs occurrence, suggesting that the COMT polymorphisms and the gene–environment interactions may alter the risk of PARIs.
Examining the Connection between Religion and Attitude toward Socio-Economic Human Rights and Attitude toward Euthanasia and Abortion among Romanian Orthodox Adolescents: Contrasting the Effects of Intrinsic and Extrinsic Religiosity
Authors: Rogobete Silviu E. , Francis Leslie J. , McKenna Ursula
Published: October 2021

This study examines the complex connection linking religion, social attitudes, and human rights in Romania, drawing on the classic distinction between extrinsic religiosity (as reflected in church attendance) and intrinsic religiosity (as reflected in personal prayer). The hypothesis that these forms of religiosity may function differently in relation to different areas of social attitudes is tested among Romanian Orthodox adolescents (N = 400), drawing on validated measures developed by the International Empirical Research Program Religion and Human Rights 2.0 to assess attitude toward socio-economic human rights and attitude toward euthanasia and abortion. In respect of attitude toward euthanasia and abortion, church attendance and personal prayer work in the same direction and with cumulative effect. Lowest acceptance of euthanasia and abortion is found among young people who attend church and pray. In respect of attitude toward socio-economic human rights, church attendance and personal prayer work in opposite directions. Frequent church attendance (extrinsic religiosity) is associated with lower endorsement of socio-economic human rights. Frequent prayer (intrinsic religiosity) is associated with higher endorsement of socio-economic human rights.
The Prediction of Running Velocity during the 30–15 Intermittent Fitness Test Using Accelerometry-Derived Metrics and Physiological Parameters: A Machine Learning Approach
Authors: Di Credico Andrea , Perpetuini David , Chiacchiaretta Piero , Cardone Daniela , Filippini Chiara , Gaggi Giulia , Merla Arcangelo , Ghinassi Barbara , Di Baldassarre Angela , Izzicupo Pascal
Published: October 2021

Measuring exercise variables is one of the most important points to consider to maximize physiological adaptations. High-intensity interval training (HIIT) is a useful method to improve both cardiovascular and neuromuscular performance. The 30–15IFT is a field test reflecting the effort elicited by HIIT, and the final velocity reached in the test is used to set the intensity of HIIT during the training session. In order to have a valid measure of the velocity during training, devices such as GPS can be used. However, in several situations (e.g., indoor setting), such devices do not provide reliable measures. The aim of the study was to predict exact running velocity during the 30–15IFT using accelerometry-derived metrics (i.e., Player Load and Average Net Force) and heart rate (HR) through a machine learning (ML) approach (i.e., Support Vector Machine) with a leave-one-subject-out cross-validation. The SVM approach showed the highest performance to predict running velocity (r = 0.91) when compared to univariate approaches using PL (r = 0.62), AvNetForce (r = 0.73) and HR only (r = 0.87). In conclusion, the presented multivariate ML approach is able to predict running velocity better than univariate ones, and the model is generalizable across subjects.
Accessibility Assessment of Buildings Based on Multi-Source Spatial Data: Taking Wuhan as a Case Study
Authors: Yang Xue , Cao Yanjia , Wu Anqi , Guo Mingqiang , Dong Zhen , Tang Luliang
Published: October 2021

The question of whether each building of housing estate has equal access to nearby social service resources (e.g., public transportation service, catering, entertainment, etc.) is a major concern of citizens. This paper takes Wuhan as a case to explore the equality in social service resource sharing of the housing estate at a microscopic level by analyzing the accessibility of each building under different travel patterns. To estimate the accessibility of each building, we developed a novel model with multi-travel modes and residential suitability evaluation of residents. The specific values of the parameters involved in the proposed model were extracted from the multi-source spatial data such as social media data, census data, point of interest, and road network data. These data were acquired from multiple platforms, e.g., Gaode map, OSM (OpenStreetMap), and GeoQ. We chose three types of districts in the city of Wuhan, including the old central district, new central district, and suburban district. We applied the proposed model to assess the accessibility of communities in these districts. Based on the results, we further analyzed whether and to what extent the distribution of each building in urban communities is equitable for social service resource sharing in China.
Induction of Apoptotic Temperature in Photothermal Therapy under Various Heating Conditions in Multi-Layered Skin Structure
Authors: Kim Donghyuk , Kim Hyunjung
Published: October 2021

Recently, photothermal therapy has attracted attention as an alternative treatment to conventional surgical techniques because it does not lead to bleeding and patients quickly recover after treatment compared to incisional surgery. Photothermal therapy induces tumor cell death through an increase in the temperature using the photothermal effect, which converts light energy into thermal energy. This study was conducted to perform numerical analysis based on heat transfer to induce apoptosis of tumor tissue under various heating conditions in photothermal therapy. The Monte Carlo method was applied to evaluate a multi-layered skin structure containing squamous cell carcinoma. Tissue-equivalent phantom experiments verified the numerical model. Based on the effective apoptosis retention ratio, the numerical analysis results showed the quantitative correlation for the laser intensity, volume fraction of gold nanorods injected into the tumor, and cooling time. This study reveals optimal conditions for maximizing apoptosis within tumor tissue while minimizing thermal damage to surrounding tissues under various heating conditions. This approach may be useful as a standard treatment when performing photothermal therapy.
Tubular Cell Cycle Response upon AKI: Revising Old and New Paradigms to Identify Novel Targets for CKD Prevention
Authors: De Chiara Letizia , Conte Carolina , Antonelli Giulia , Lazzeri Elena
Published: October 2021

Acute kidney injury (AKI) is characterized by a rapid deterioration of kidney function, representing a global healthcare concern. In addition, AKI survivors frequently develop chronic kidney disease (CKD), contributing to a substantial proportion of disease burden globally. Yet, over the past 30 years, the burden of CKD has not declined to the same extent as many other important non-communicable diseases, implying a substantial deficit in the understanding of the disease progression. The assumption that the kidney response to AKI is based on a high proliferative potential of proximal tubular cells (PTC) caused a critical confounding factor, which has led to a limited development of strategies to prevent AKI and halt progression toward CKD. In this review, we discuss the latest findings on multiple mechanisms of response related to cell cycle behavior of PTC upon AKI, with a specific focus on their biological relevance. Collectively, we aim to (1) provide a new perspective on interpreting cell cycle progression of PTC in response to damage and (2) discuss how this knowledge can be used to choose the right therapeutic window of treatment for preserving kidney function while avoiding CKD progression.
Interleukin-15 in Outcomes of Pregnancy
Authors: Gordon Scott M.
Published: October 2021

Interleukin-15 (IL-15) is a pleiotropic cytokine that classically acts to support the development, maintenance, and function of killer lymphocytes. IL-15 is abundant in the uterus prior to and during pregnancy, but it is subject to tight spatial and temporal regulation. Both mouse models and human studies suggest that homeostasis of IL-15 is essential for healthy pregnancy. Dysregulation of IL-15 is associated with adverse outcomes of pregnancy. Herein, we review producers of IL-15 and responders to IL-15, including non-traditional responders in the maternal uterus and fetal placenta. We also review regulation of IL-15 at the maternal–fetal interface and propose mechanisms of action of IL-15 to facilitate additional study of this critical cytokine in the context of pregnancy.
BET Proteins as Attractive Targets for Cancer Therapeutics
Authors: Sarnik Joanna , Popławski Tomasz , Tokarz Paulina
Published: October 2021

Transcriptional dysregulation is a hallmark of cancer and can be an essential driver of cancer initiation and progression. Loss of transcriptional control can cause cancer cells to become dependent on certain regulators of gene expression. Bromodomain and extraterminal domain (BET) proteins are epigenetic readers that regulate the expression of multiple genes involved in carcinogenesis. BET inhibitors (BETis) disrupt BET protein binding to acetylated lysine residues of chromatin and suppress the transcription of various genes, including oncogenic transcription factors. Phase I and II clinical trials demonstrated BETis’ potential as anticancer drugs against solid tumours and haematological malignancies; however, their clinical success was limited as monotherapies. Emerging treatment-associated toxicities, drug resistance and a lack of predictive biomarkers limited BETis’ clinical progress. The preclinical evaluation demonstrated that BETis synergised with different classes of compounds, including DNA repair inhibitors, thus supporting further clinical development of BETis. The combination of BET and PARP inhibitors triggered synthetic lethality in cells with proficient homologous recombination. Mechanistic studies revealed that BETis targeted multiple essential homologous recombination pathway proteins, including RAD51, BRCA1 and CtIP. The exact mechanism of BETis’ anticancer action remains poorly understood; nevertheless, these agents provide a novel approach to epigenome and transcriptome anticancer therapy.
The Expression and Localisation of G-Protein-Coupled Inwardly Rectifying Potassium (GIRK) Channels Is Differentially Altered in the Hippocampus of Two Mouse Models of Alzheimer’s Disease
Authors: Alfaro-Ruiz Rocío , Martín-Belmonte Alejandro , Aguado Carolina , Hernández Félix , Moreno-Martínez Ana Esther , Ávila Jesús , Luján Rafael
Published: October 2021

G protein-gated inwardly rectifying K+ (GIRK) channels are the main targets controlling excitability and synaptic plasticity on hippocampal neurons. Consequently, dysfunction of GIRK-mediated signalling has been implicated in the pathophysiology of Alzheimer´s disease (AD). Here, we provide a quantitative description on the expression and localisation patterns of GIRK2 in two transgenic mice models of AD (P301S and APP/PS1 mice), combining histoblots and immunoelectron microscopic approaches. The histoblot technique revealed differences in the expression of GIRK2 in the two transgenic mice models. The expression of GIRK2 was significantly reduced in the hippocampus of P301S mice in a laminar-specific manner at 10 months of age but was unaltered in APP/PS1 mice at 12 months compared to age-matched wild type mice. Ultrastructural approaches using the pre-embedding immunogold technique, demonstrated that the subcellular localisation of GIRK2 was significantly reduced along the neuronal surface of CA1 pyramidal cells, but increased in its frequency at cytoplasmic sites, in both P301S and APP/PS1 mice. We also found a decrease in plasma membrane GIRK2 channels in axon terminals contacting dendritic spines of CA1 pyramidal cells in P301S and APP/PS1 mice. These data demonstrate for the first time a redistribution of GIRK channels from the plasma membrane to intracellular sites in different compartments of CA1 pyramidal cells. Altogether, the pre- and post-synaptic reduction of GIRK2 channels suggest that GIRK-mediated alteration of the excitability in pyramidal cells could contribute to the cognitive dysfunctions as described in the two AD animal models.
Increased Expression of N2BA Titin Corresponds to More Compliant Myofibrils in Athlete’s Heart
Authors: Kellermayer Dalma , Kiss Bálint , Tordai Hedvig , Oláh Attila , Granzier Henk L. , Merkely Béla , Kellermayer Miklós , Radovits Tamás
Published: October 2021

Long-term exercise induces physiological cardiac adaptation, a condition referred to as athlete’s heart. Exercise tolerance is known to be associated with decreased cardiac passive stiffness. Passive stiffness of the heart muscle is determined by the giant elastic protein titin. The adult cardiac muscle contains two titin isoforms: the more compliant N2BA and the stiffer N2B. Titin-based passive stiffness may be controlled by altering the expression of the different isoforms or via post-translational modifications such as phosphorylation. Currently, there is very limited knowledge about titin’s role in cardiac adaptation during long-term exercise. Our aim was to determine the N2BA/N2B ratio and post-translational phosphorylation of titin in the left ventricle and to correlate the changes with the structure and transverse stiffness of cardiac sarcomeres in a rat model of an athlete’s heart. The athlete’s heart was induced by a 12-week-long swim-based training. In the exercised myocardium the N2BA/N2B ratio was significantly increased, Ser11878 of the PEVK domain was hypophosphorlyated, and the sarcomeric transverse elastic modulus was reduced. Thus, the reduced passive stiffness in the athlete’s heart is likely caused by a shift towards the expression of the longer cardiac titin isoform and a phosphorylation-induced softening of the PEVK domain which is manifested in a mechanical rearrangement locally, within the cardiac sarcomere.
ASP-Enzymosomes with Saccharomyces cerevisiae Asparaginase II Expressed in Pichia pastoris: Formulation Design and In Vitro Studies of a Potential Antileukemic Drug
Authors: Girão Luciana F. C. , Carvalheiro Manuela Colla , Ferreira-Silva Margarida , da Rocha Surza L. G. , Perales Jonas , Martins M. Bárbara F. , Ferrara Maria Antonieta , Bon Elba P. S. , Corvo M. Luísa
Published: October 2021

The bacterial enzyme asparaginase is the main treatment option for acute lymphoblastic leukemia. However, it causes side effects, such as immunological reactions, and presents undesirable glutaminase activity. As an alternative, we have been studying asparaginase II from Saccharomyces cerevisiae, coded by ASP3 gene, which was cloned and expressed in Pichia pastoris. The recombinant asparaginase (ASP) presented antileukemic activity and a glutaminase activity 100 times lower in comparison to its asparaginase activity. In this work, we describe the development of a delivery system for ASP via its covalent attachment to functionalized polyethylene glycol (PEG) polymer chains in the outer surface of liposomes (ASP-enzymosomes). This new delivery system demonstrated antiproliferative activity against K562 (chronic myeloid leukemia) and Jurkat (acute lymphocytic leukemia) cell lines similar to that of ASP. The antiproliferative response of the ASP-enzymosomes against the Jurkat cells suggests equivalence to that of the free Escherichia coli commercial asparaginase (Aginasa®). Moreover, the ASP-enzymosomes were stable at 4 °C with no significant loss of activity within 4 days and retained 82% activity up to 37 days. Therefore, ASP-enzymosomes are a promising antileukemic drug.
Bee Venom Melittin Disintegrates the Respiration of Mitochondria in Healthy Cells and Lymphoblasts, and Induces the Formation of Non-Bilayer Structures in Model Inner Mitochondrial Membranes
Authors: Gasanoff Edward , Liu Yipeng , Li Feng , Hanlon Paul , Garab Győző
Published: October 2021

In this paper, we examined the effects of melittin, a bee venom membrane-active peptide, on mitochondrial respiration and cell viability of healthy human lymphocytes (HHL) and Jurkat cells, as well as on lymphoblasts from acute human T cell leukemia. The viability of melittin-treated cells was related to changes in O2 consumption and in the respiratory control index (RCI) of mitochondria isolated from melittin-pretreated cells as well as of mitochondria first isolated from cells and then directly treated with melittin. It was shown that melittin is three times more cytotoxic to Jurkat cells than to HHL, but O2 consumption and RCI values of mitochondria from both cell types were equally affected by melittin when melittin was directly added to mitochondria. To elucidate the molecular mechanism of melittin’s cytotoxicity to healthy and cancer cells, the effects of melittin on lipid-packing and on the dynamics in model plasma membranes of healthy and cancer cells, as well as of the inner mitochondrial membrane, were studied by EPR spin probes. The affinity of melittin binding to phosphatidylcholine, phosphatidylserine, phosphatidic acid and cardiolipin, and binding sites of phospholipids on the surface of melittin were studied by 31P-NMR, native PAGE and AutoDock modeling. It is suggested that the melittin-induced decline of mitochondrial bioenergetics contributes primarily to cell death; the higher cytotoxicity of melittin to cancer cells is attributed to its increased permeability through the plasma membrane.
DRP1-Mediated Mitochondrial Fission Regulates Lung Epithelial Response to Allergen
Authors: Bruno Sierra R. , Kumar Amit , Mark Zoe F. , Chandrasekaran Ravishankar , Nakada Emily , Chamberlain Nicolas , Mihavics Bethany , Walzer Joseph , Cahoon Jonathon , Dixon Anne E. , Cunniff Brian , Anathy Vikas
Published: October 2021

Mitochondria regulate a myriad of cellular functions. Dysregulation of mitochondrial control within airway epithelial cells has been implicated in the pro-inflammatory response to allergens in asthma patients. Because of their multifaceted nature, mitochondrial structure must be tightly regulated through fission and fusion. Dynamin Related Protein 1 (DRP1) is a key driver of mitochondrial fission. During allergic asthma, airway epithelial mitochondria appear smaller and structurally altered. The role of DRP1-mediated mitochondrial fission, however, has not been fully elucidated in epithelial response to allergens. We used a Human Bronchial Epithelial Cell line (HBECs), primary Mouse Tracheal Epithelial Cells (MTECs), and conditional DRP1 ablation in lung epithelial cells to investigate the impact of mitochondrial fission on the pro-inflammatory response to house dust mite (HDM) in vitro and in vivo. Our data suggest that, following HDM challenge, mitochondrial fission is rapidly upregulated in airway epithelial cells and precedes production of pro-inflammatory cytokines and chemokines. Further, deletion of Drp1 in lung epithelial cells leads to decreased fission and enhanced pro-inflammatory signaling in response to HDM in vitro, as well as enhanced airway hyper-responsiveness (AHR), inflammation, differential mucin transcription, and epithelial cell death in vivo. Mitochondrial fission, therefore, regulates the lung epithelial pro-inflammatory response to HDM.
Extended Cheese Whey Fermentation Produces a Novel Casein-Derived Antibacterial Polypeptide That Also Inhibits Gelatinases MMP-2 and MMP-9
Authors: Santos Maria Isabel , Lima Ana , Mota Joana , Rebelo Patrícia , Ferreira Ricardo Boavida , Pedroso Laurentina , Ferreira Maria Adélia , Sousa Isabel
Published: October 2021

Our previous works produced a whey fermentation methodology that yielded antibacterial activity and potential inhibition of matrix metalloproteases (MMP)-2 and -9. Here, we evaluated if these activities were due to fermentation-produced peptides. Prolonged fermentation was carried out in the presence of our specific lactic acid bacteria (LAB) consortium. LAB fermentation yielded a total of 11 polypeptides, which were predominantly produced after 6 days of fermentation. One which was derived from beat casein presented a particularly high antibacterial activity against food pathogenic bacteria and was more effective than standard food disinfectants. This polypeptide was further studied and was also found to be active against several strains of pathogenic bacteria, including methicillin-resistant Staphylococcus aureus (MRSA), in a dose-dependent manner. It also inhibited MMP-2 and MMP-9 whilst reducing HT29 cancer cell migration in vitro. Overall, this novel whey-derived polypeptide presents dual antibacterial and anti-inflammatory activity, revealing a strong potential to be used in functional foods or as a nutraceutical. Its identification and further characterization can open novel perspectives in the field of preventive/curative diets related to gut microbiota, gut inflammation, and cancer prevention, particularly if used in in vivo studies.
CsCDPK6, a CsSAMS1-Interacting Protein, Affects Polyamine/Ethylene Biosynthesis in Cucumber and Enhances Salt Tolerance by Overexpression in Tobacco
Authors: Zhu Heyuan , He Meiwen , Jahan Mohammad , Wu Jianqiang , Gu Qinsheng , Shu Sheng , Sun Jin , Guo Shirong
Published: October 2021

S-adenosylmethionine synthetase (SAMS) plays a crucial role in regulating stress responses. In a recent study, we found that overexpression of the cucumber gene CsSAMS1 in tobacco can affect the production of polyamines and ethylene, as well as enhancing the salt stress tolerance of tobacco, but the exact underlying mechanisms are elusive. The calcium-dependent protein kinase (CDPK) family is ubiquitous in plants and performs different biological functions in plant development and response to abiotic stress. We used a yeast two-hybrid system to detect whether the protein CDPK6 could interact with SAMS1 and verified their interaction by bimolecular fluorescence complementation (BiFC) and co-immunoprecipitation (Co-IP) assays. To further explore the function of cucumber CDPK6, we isolated and characterized CsCDPK6 in cucumber. CsCDPK6 is a membrane protein that is highly expressed under various abiotic stresses, including salt stress. It was also observed that ectopic overexpression of CsCDPK6 in tobacco enhanced salt tolerance. Under salt stress, CsCDPK6-overexpressing lines enhanced the survival rate and reduced stomatal apertures in comparison to wild-type (WT) lines, as well as lowering malondialdehyde (MDA) and hydrogen peroxide (H2O2) contents and causing less relative electrolyte leakage. Moreover, repression of CsCDPK6 expression by virus-induced gene silencing (VIGS) in cucumber seedling cotyledons under salt stress increased ethylene production and promoted the transformation from putrescine (Put) to spermidine (Spd) and spermine (Spm). These findings shed light on the interaction of CsSAMS1 and CsCDPK6, which functions positively to regulate salt stress in plants.
A Small-Scale Analysis of Elevational Species Richness and Beta Diversity Patterns of Arthropods on an Oceanic Island (Terceira, Azores)
Authors: de Vries Jan Peter Reinier , van Loon Emiel , Borges Paulo A. V.
Published: October 2021

We present an analysis of arthropod diversity patterns in native forest communities along the small elevation gradient (0–1021 m a.s.l.) of Terceira island, Azores (Portugal). We analysed (1) how the alpha diversity of Azorean arthropods responds to increasing elevation and (2) differs between endemic, native non-endemic and introduced (alien) species, and (3) the contributions of species replacement and richness difference to beta diversity. Arthropods were sampled using SLAM traps between 2014 and 2018. We analysed species richness indicators, the Hill series and beta diversity partitioning (species replacement and species richness differences). Selected orders (Araneae, Coleoptera, Hemiptera and Psocoptera) and endemic, native non-endemic and introduced species were analysed separately. Total species richness shows a monotonic decrease with elevation for all species and Coleoptera and Psocoptera, but peaks at mid-high elevation for Araneae and endemic species. Introduced species richness decreases strongly with elevation especially. These patterns are most likely driven by climatic factors but also influenced by human disturbance. Beta diversity is, for most groups, the main component of total (gamma) diversity along the gradient but shows no relation with elevation. It results from a combined effect of richness decrease with elevation and species replacement in groups with many narrow-ranged species.
The Structure of the Endophallus Is a New Promising Feature and a Key to Study of Taxonomy of the Subgenus Metallotimarcha of the Genus Timarcha (Coleoptera, Chrysomelidae) in the Caucasus
Authors: Bieńkowski Andrzej
Published: October 2021

A comparative morphological study of the members of the subgenus Metallotimarcha revealed that Caucasian Timarcha hummelii is a separate species, which differs from European members of the subgenus (T. metallica, T. corinthia, T. gibba) by the internal structure of male aedeagus, namely, manubrium of the endophallus. Morphology of manubrium is described for all species of the subgenus. External characters and the shape of male aedeagus do not permit separating the members of the subgenus distinctly. Examination of both external characters and endophallus structure in the specimens from the Caucasus revealed that T. armeniaca and T. hummelii starcki are the synonyms of T. hummelii hummelii, but not separate species or subspecies. Neotypes of T. hummelii and T. armeniaca are designated. Syntypes of T. hummelii starcki are examined. The key to the species for the subgenus Metallotimarcha is compiled.
Full-Length SMRT Transcriptome Sequencing and SSR Analysis of Bactrocera dorsalis (Hendel)
Authors: Ouyang Huili , Wang Xiaoyun , Zheng Xialin , Lu Wen , Qin Fengping , Chen Chao
Published: October 2021

Bactrocera dorsalis (Hendel), as one of the most notorious and destructive invasive agricultural pests in the world, causes damage to over 250 different types of fruits and vegetables throughout tropical and subtropical areas. PacBio single-molecule real-time (SMRT) sequencing was used to generate the full-length transcriptome data of B. dorsalis. A total of 40,319,890 subreads (76.6 Gb, clean reads) were generated, including 535,241 circular consensus sequences (CCSs) and 386,916 full-length non-concatemer reads (FLNCs). Transcript cluster analysis of the FLNC reads revealed 22,780 high-quality reads (HQs). In total, 12,274 transcripts were functionally annotated based on four different databases. A total of 1978 SSR loci were distributed throughout 1714 HQ transcripts, of which 1926 were complete SSRs and 52 were complex SSRs. Among the total SSR loci, 2–3 nucleotide repeats were dominant, occupying 83.62%, of which di- and tri- nucleotide repeats were 39.38% and 44.24%, respectively. We detected 105 repeat motifs, of which AT/AT (50.19%), AC/GT (39.15%), CAA/TTG (32.46%), and ACA/TGT (10.86%) were the most common in di- and tri-nucleotide repeats. The repeat SSR motifs were 12–190 bp in length, and 1638 (88.02%) were shorter than 20 bp. According to the randomly selected microsatellite sequence, 80 pairs of primers were designed, and 174 individuals were randomly amplified by PCR using primers. The number of primers that had amplification products with clear bands and showed good polymorphism came to 41, indicating that this was a feasible way to explore SSR markers from the transcriptomic data of B. dorsalis. These results lay a foundation for developing highly polymorphic microsatellites for researching the functional genomics, population genetic structure, and genetic diversity of B. dorsalis.
Hereditary Hemorrhagic Telangiectasia: Success of the Osler Calendar for Documentation of Treatment and Course of Disease
Authors: Seebauer Caroline T. , Freigang Viola , Schwan Franziska E. , Fischer René , Bohr Christopher , Kühnel Thomas S. , Andorfer Kornelia E. C.
Published: October 2021

Hereditary hemorrhagic telangiectasia (HHT; Rendu-Osler-Weber syndrome) affects the capillary and larger vessels, leading to arteriovenous shunts. Epistaxis is the main symptom impairing quality of life. The aim of the Osler Calendar is to offer information about the extent of the systemic disease and the current state of treatment. A care plan with information on the rare disease and self-treatment of epistaxis was created. Organ examinations and ongoing treatments were recorded. A questionnaire documents the treatment success, including patient satisfaction, frequency of hemorrhage and hemoglobin levels. The patients using the Osler Calendar for at least one year (n = 54) were surveyed. Eighty-five percent of patients (n = 46) used the calendar to gain information about HHT. Seventy-two percent (n = 39) used the Osler Calendar for instructions on the self-treatment of nosebleeds. The calendar increased patients’ understanding for the need for organ screenings from 48% (n = 26) to 81% (n = 44). Seventy-nine percent (n = 43) of patients confirmed that the Osler Calendar documented their therapeutic process either well or very well. Fifty-two percent (n = 28) saw an improvement in the therapeutic process due to the documentation. The Osler Calendar records the individual intensity of the disease and facilitates the communication between attending physicians. It is a tool for specialists to review treatment strategies. Furthermore, the calendar enhances patients’ comprehension of their condition.
Autoimmunity in Primary Immunodeficiency Disorders: An Updated Review on Pathogenic and Clinical Implications
Authors: Costagliola Giorgio , Cappelli Susanna , Consolini Rita
Published: October 2021

During the last years, studies investigating the intriguing association between immunodeficiency and autoimmunity led to the discovery of new monogenic disorders, the improvement in the knowledge of the pathogenesis of autoimmunity, and the introduction of targeted treatments. Autoimmunity is observed with particular frequency in patients with primary antibody deficiencies, such as common variable immunodeficiency (CVID) and selective IgA deficiency, but combined immunodeficiency disorders (CIDs) and disorders of innate immunity have also been associated with autoimmunity. Among CIDs, the highest incidence of autoimmunity is described in patients with autoimmune polyendocrine syndrome 1, LRBA, and CTLA-4 deficiency, and in patients with STAT-related disorders. The pathogenesis of autoimmunity in patients with immunodeficiency is far to be fully elucidated. However, altered germ center reactions, impaired central and peripheral lymphocyte negative selection, uncontrolled lymphocyte proliferation, ineffective cytoskeletal function, innate immune defects, and defective clearance of the infectious agents play an important role. In this paper, we review the main immunodeficiencies associated with autoimmunity, focusing on the pathogenic mechanisms responsible for autoimmunity in each condition and on the therapeutic strategies. Moreover, we provide a diagnostic algorithm for the diagnosis of PIDs in patients with autoimmunity.
How to Assess the Efficacy of Interventions for Actinic Keratosis? A Review with a Focus on Long-Term Results
Authors: Steeb Theresa , Wessely Anja , Petzold Anne , Schmitz Lutz , Dirschka Thomas , Berking Carola , Heppt Markus V.
Published: October 2021

Actinic keratoses (AK) are common lesions of the skin caused by cumulative sun exposure. Since AK may progress to invasive cutaneous squamous cell carcinoma (cSCC), guidelines uniformly recommend early and consequent treatment. A variety of interventions are available; however, most randomized controlled trials, meta-analyses, and guidelines focus on outcomes that are usually evaluated 8–12 weeks after the end of treatment. Importantly, these assessments can capture the short-term, transient outcomes, but do not allow any conclusions about long-term results to be drawn and do not reflect the probability of transition towards cSCC. Until now, few studies have assessed the long-term results of interventions for AK. Indeed, finding the most appropriate end-point and adjunct time point for determining the long-term results of interventions for AK remains a challenge. Here, we provide an overview of the different ways of measuring the efficacy of AK treatments, such as using recurrence rates or sustained clearance rates, and discuss methodological aspects. Furthermore, we highlight the importance of evidence from post-marketing surveillance trials for the detection of efficacy values and safety signals. Additionally, we emphasize that a follow-up period of 12 months might not be sufficient to reflect the long-term results and stress the urgent need for a longer follow-up period and regular risk-stratified surveillance.