bluuebunny/crossref_metadata_2025 huggingface dataset:
The Impact of Parental Incarceration on Sport Participation Trajectories from Adolescence to Young Adulthood
Authors: Park Ji-Won , Hwang Jongnam , Lee Chung Gun , Ahn Hyoyeon , Kim Hanbeom
Published: July 2020

Given the potential negative effects of parental incarceration on millions of people, it is critical to examine the possible short- and long-term effects of parental incarceration on individuals. This study examines the effect of parental incarceration on the sports participation trajectories of children ranging from adolescence to young adulthood. Group-based trajectory models were set up using SAS analytics software to examine how parental incarceration affects the sports participation trajectories of children from adolescence to young adulthood. Data were drawn from the first four waves of the National Longitudinal Study of Adolescent Health in the United States (N = 6504). Neither paternal nor maternal incarceration had any significant effect on the trajectories of male participants. On the other hand, females who experienced father incarceration were more likely to be in the low-stable versus high-decreasing group (coefficient = −0.721, p < 0.05). Based on the results of this study, we conclude that programs promoting sports participation are needed for females who have experienced paternal incarceration. The results of this study also suggest that group-based trajectory modeling is a useful technique to examine the trajectories of sports participation from adolescence through to young adulthood.
Return to Work and Quality of Life after Stroke in Italy: A Study on the Efficacy of Technologically Assisted Neurorehabilitation
Authors: Ghanbari Ghoshchi Sheyda , De Angelis Sara , Morone Giovanni , Panigazzi Monica , Persechino Benedetta , Tramontano Marco , Capodaglio Edda , Zoccolotti Pierluigi , Paolucci Stefano , Iosa Marco
Published: July 2020

Cerebrovascular diseases, including stroke, are historically considered diseases of old adults so only in a few studies has “return to work” (RTW) been considered as an index of rehabilitative outcome. At the moment, data on RTW in patients with stroke are highly variable: four different reviews reported the following ranges: 11–85%, 19–73%, 22–53%, and 40–45%. The absence of re-integration to work after a stroke is shown to be associated with an increase of cardiac disorders and depression, with a higher level of mortality, with social isolation and with insufficient adaptive skills. The aim of this study was to verify the effectiveness of technological treatment, performed with optic (SonicHand) and wearable (Riablo™) systems providing auditory and visual biofeedback, on RTW in patients with stroke. RTW was found to be associated with a higher independence in the activities of daily living (assessed by the Modified Barthel Index). No significant differences were found between technological versus conventional rehabilitation in terms of RTW, despite the former showing a higher odds ratio than the latter (OR = 9 vs. 6). Assistive devices were mainly used in patients who had not returned to work. Finally, quality of life was found higher in those patients who returned to work with the same conditions (work duties and time) as before stroke.
Relationships among Leisure Physical Activity, Sedentary Lifestyle, Physical Fitness, and Happiness in Adults 65 Years or Older in Taiwan
Authors: Lin Yi-Tien , Chen Mingchih , Ho Chien-Chang , Lee Tian-Shyug
Published: July 2020

The purpose of this study is to understand the relationship among leisure physical activity, sedentary lifestyle, physical fitness, and happiness in healthy elderly adults aged over 65 years old in Taiwan. Data were recruited from the National Physical Fitness Survey in Taiwan, which was proposed in the Project on the Establishment of Physical Fitness Testing Stations by the Sports Administration of the Ministry of Education. Participants were recruited from fitness testing stations set up in 22 counties and cities from October 2015 to May 2016. A total of 20,111 healthy older adults aged 65–102 years were recruited as research participants. The fitness testing procedure was described to all participants, who were provided with a standardized structured questionnaire. Participants’ data included sex, city or county of residence, living status (living together with others or living alone), education level, and income. Physical fitness testing was conducted in accordance with The Fitness Guide for Older Adults published by the Sports Administration of the Ministry of Education. The testing involved cardiorespiratory endurance, muscle strength, muscle endurance, flexibility, balance, and body composition. The t-test was used to evaluate the differences between continuous and grade variables under the two classification variables of sex, city or county of residence, and living status. We used the MARS (multivariate adaptive regression splines) model to analyze the effects of physical fitness variables and leisure physical activity variables on happiness. Among healthy elderly adults, sex, age, living status, body mass index, and leisure physical activity habits proved to be related to happiness. Aerobic endurance (2-min step test), muscular strength and endurance (30-s arm curl and 30-s chair stand tests), flexibility (back stretch and chair sit-and-reach tests), and balance ability (8-foot up-and-go tests and one-leg stance with eyes open tests) were found to be related to happiness. The results of this study indicate that increased physical activity and intensity, as well as physical fitness performance, are associated with improved happiness.
Driving Accidents, Driving Violations, Symptoms of Attention-Deficit-Hyperactivity (ADHD) and Attentional Network Tasks
Authors: Zamani Sani Seyed Hojjat , Fathirezaie Zahra , Sadeghi-Bazargani Homayoun , Badicu Georgian , Ebrahimi Safyeh , Grosz Robert Wilhelm , Sadeghi Bahmani Dena , Brand Serge
Published: July 2020

Background: Iran has serious problems with traffic-related injuries and death. A major reason for traffic accidents is cognitive failure due to deficits in attention. In this study, we investigated the associations between traffic violations, traffic accidents, symptoms of attention-deficit/hyperactivity disorder (ADHD), age, and on an attentional network task in a sample of Iranian adults. Methods: A total of 274 participants (mean age: 31.37 years; 80.7% males) completed questionnaires covering demographic information, driving violations, traffic accidents, and symptoms of ADHD. In addition, they underwent an objective attentional network task (ANT), based on Posner’s concept of attentional networks. Results: More frequent traffic violations, correlated with lower age and poorer performance on the attentional network tasks. Higher symptoms of ADHD were associated with more accidents and more traffic violations, but not with the performance of the attentional tasks. Higher ADHD scores, a poorer performance on attentional network tasks, and younger age predicted traffic violations. Only higher symptoms of ADHD predicted more traffic accidents. Conclusions: In a sample of Iranian drivers, self-rated symptoms of ADHD appeared to be associated with traffic violations and accidents, while symptoms of ADHD were unrelated to objectively assessed performance on an attentional network task. Poor attentional network performance was a significant predictor of traffic violations but not of accidents. To increase traffic safety, both symptoms of ADHD and attentional network performance appear to merit particular attention.
Population Trends and Urbanization: Simulating Density Effects Using a Local Regression Approach
Authors: Polinesi Gloria , Recchioni Maria Cristina , Turco Rosario , Salvati Luca , Rontos Kostas , Rodrigo-Comino Jesús , Benassi Federico
Published: July 2020

Density-dependent population growth regulates long-term urban expansion and shapes distinctive socioeconomic trends. Despite a marked heterogeneity in the spatial distribution of the resident population, Mediterranean European countries are considered more homogeneous than countries in other European regions as far as settlement structure and processes of metropolitan growth are concerned. However, rising socioeconomic inequalities among Southern European regions reflect latent demographic and territorial transformations that require further investigation. An integrated assessment of the spatio-temporal distribution of resident populations in more than 1000 municipalities (1961–2011) was carried out in this study to characterize density-dependent processes of metropolitan growth in Greece. Using geographically weighted regressions, the results of our study identified distinctive local relationships between population density and growth rates over time. Our results demonstrate that demographic growth rates were non-linearly correlated with other variables, such as population density, with positive and negative impacts during the first (1961–1971) and the last (2001–2011) observation decade, respectively. These findings outline a progressive shift over time from density-dependent processes of population growth, reflecting a rapid development of large metropolitan regions (Athens, Thessaloniki) in the 1960s, to density-dependent processes more evident in medium-sized cities and accessible rural regions in the 2000s. Density-independent processes of population growth have been detected in the intermediate study period (1971–2001). This work finally discusses how a long-term analysis of demographic growth, testing for density-dependent mechanisms, may clarify the intrinsic role of population concentration and dispersion in different phases of the metropolitan cycle in Mediterranean Europe.
Modeling Major Rural Land-Use Changes Using the GIS-Based Cellular Automata Metronamica Model: The Case of Andalusia (Southern Spain)
Authors: Navarro Cerrillo Rafael M. , Palacios Rodríguez Guillermo , Clavero Rumbao Inmaculada , Lara Miguel Ángel , Bonet Francisco Javier , Mesas-Carrascosa Francisco-Javier
Published: July 2020

The effective and efficient planning of rural land-use changes and their impact on the environment is critical for land-use managers. Many land-use growth models have been proposed for forecasting growth patterns in the last few years. In this work; a cellular automata (CA)-based land-use model (Metronamica) was tested to simulate (1999–2007) and predict (2007–2035) land-use dynamics and land-use changes in Andalucía (Spain). The model was calibrated using temporal changes in land-use covers and was evaluated by the Kappa index. GIS-based maps were generated to study major rural land-use changes (agriculture and forests). The change matrix for 1999–2007 showed an overall area change of 674971 ha. The dominant land uses in 2007 were shrubs (30.7%), woody crops on dry land (17.3%), and herbaceous crops on dry land (12.7%). The comparison between the reference and the simulated land-use maps of 2007 showed a Kappa index of 0.91. The land-cover map for the projected PRELUDE scenarios provided the land-cover characteristics of 2035 in Andalusia; developed within the Metronamica model scenarios (Great Escape; Evolved Society; Clustered Network; Lettuce Surprise U; and Big Crisis). The greatest differences were found between Great Escape and Clustered Network and Lettuce Surprise U. The observed trend (1999–2007–2035) showed the greatest similarity with the Big Crisis scenario. Land-use projections facilitate the understanding of the future dynamics of land-use change in rural areas; and hence the development of more appropriate plans and policies
Inter-Regulation of Kv4.3 and Voltage-Gated Sodium Channels Underlies Predisposition to Cardiac and Neuronal Channelopathies
Authors: Clatot Jérôme , Neyroud Nathalie , Cox Robert , Souil Charlotte , Huang Jing , Guicheney Pascale , Antzelevitch Charles
Published: July 2020

Background: Genetic variants in voltage-gated sodium channels (Nav) encoded by SCNXA genes, responsible for INa, and Kv4.3 channels encoded by KCND3, responsible for the transient outward current (Ito), contribute to the manifestation of both Brugada syndrome (BrS) and spinocerebellar ataxia (SCA19/22). We examined the hypothesis that Kv4.3 and Nav variants regulate each other’s function, thus modulating INa/Ito balance in cardiomyocytes and INa/I(A) balance in neurons. Methods: Bicistronic and other constructs were used to express WT or variant Nav1.5 and Kv4.3 channels in HEK293 cells. INa and Ito were recorded. Results: SCN5A variants associated with BrS reduced INa, but increased Ito. Moreover, BrS and SCA19/22 KCND3 variants associated with a gain of function of Ito, significantly reduced INa, whereas the SCA19/22 KCND3 variants associated with a loss of function (LOF) of Ito significantly increased INa. Auxiliary subunits Navβ1, MiRP3 and KChIP2 also modulated INa/Ito balance. Co-immunoprecipitation and Duolink studies suggested that the two channels interact within the intracellular compartments and biotinylation showed that LOF SCN5A variants can increase Kv4.3 cell-surface expression. Conclusion: Nav and Kv4.3 channels modulate each other’s function via trafficking and gating mechanisms, which have important implications for improved understanding of these allelic cardiac and neuronal syndromes.
Genetic Dissection and Identification of Candidate Genes for Salinity Tolerance Using Axiom®CicerSNP Array in Chickpea
Authors: Soren Khela Ram , Madugula Praveen , Kumar Neeraj , Barmukh Rutwik , Sengar Meenu Singh , Bharadwaj Chellapilla , Sharma Parbodh Chander , Singh Sarvjeet , Bhandari Aditi , Singh Jogendra , Sanwal Satish Kumar , Pal Madan , P.R. Sneha Priya , Mann Anita , Sagurthi Someswar Rao , PS Shanmugavadivel , Siddique Kadambot H.M. , Singh Narendra Pratap , Roorkiwal Manish , Varshney Rajeev K
Published: July 2020

Globally, chickpea production is severely affected by salinity stress. Understanding the genetic basis for salinity tolerance is important to develop salinity tolerant chickpeas. A recombinant inbred line (RIL) population developed using parental lines ICCV 10 (salt-tolerant) and DCP 92-3 (salt-sensitive) was screened under field conditions to collect information on agronomy, yield components, and stress tolerance indices. Genotyping data generated using Axiom®CicerSNP array was used to construct a linkage map comprising 1856 SNP markers spanning a distance of 1106.3 cM across eight chickpea chromosomes. Extensive analysis of the phenotyping and genotyping data identified 28 quantitative trait loci (QTLs) explaining up to 28.40% of the phenotypic variance in the population. We identified QTL clusters on CaLG03 and CaLG06, each harboring major QTLs for yield and yield component traits under salinity stress. The main-effect QTLs identified in these two clusters were associated with key genes such as calcium-dependent protein kinases, histidine kinases, cation proton antiporter, and WRKY and MYB transcription factors, which are known to impart salinity stress tolerance in crop plants. Molecular markers/genes associated with these major QTLs, after validation, will be useful to undertake marker-assisted breeding for developing better varieties with salinity tolerance.
Cells Derived from Human Long Bone Appear More Differentiated and More Actively Stimulate Osteoclastogenesis Compared to Alveolar Bone-Derived Cells
Authors: Kelder Cindy , Kleverlaan Cornelis J. , Gilijamse Marjolijn , Bakker Astrid D. , de Vries Teun J.
Published: July 2020

Osteoblasts derived from mouse skulls have increased osteoclastogenic potential compared to long bone osteoblasts when stimulated with 1,25(OH)2 vitamin D3 (vitD3). This indicates that bone cells from specific sites can react differently to biochemical signals, e.g., during inflammation or as emitted by bioactive bone tissue-engineering constructs. Given the high turn-over of alveolar bone, we hypothesized that human alveolar bone-derived osteoblasts have an increased osteogenic and osteoclastogenic potential compared to the osteoblasts derived from long bone. The osteogenic and osteoclastogenic capacity of alveolar bone cells and long bone cells were assessed in the presence and absence of osteotropic agent vitD3. Both cell types were studied in osteogenesis experiments, using an osteogenic medium, and in osteoclastogenesis experiments by co-culturing osteoblasts with peripheral blood mononuclear cells (PBMCs). Both osteogenic and osteoclastic markers were measured. At day 0, long bones seem to have a more late-osteoblastic/preosteocyte-like phenotype compared to the alveolar bone cells as shown by slower proliferation, the higher expression of the matrix molecule Osteopontin (OPN) and the osteocyte-enriched cytoskeletal component Actin alpha 1 (ACTA1). This phenotype was maintained during the osteogenesis assays, where long bone-derived cells still expressed more OPN and ACTA1. Under co-culture conditions with PBMCs, long bone cells also had a higher Tumor necrose factor-alfa (TNF-α) expression and induced the formation of osteoclasts more than alveolar bone cells. Correspondingly, the expression of osteoclast genes dendritic cell specific transmembrane protein (DC-STAMP) and Receptor activator of nuclear factor kappa-Β ligand (RankL) was higher in long bone co-cultures. Together, our results indicate that long bone-derived osteoblasts are more active in bone-remodeling processes, especially in osteoclastogenesis, than alveolar bone-derived cells. This indicates that tissue-engineering solutions need to be specifically designed for the site of application, such as defects in long bones vs. the regeneration of alveolar bone after severe periodontitis.
HLA-G Expression in Human Mesenchymal Stem Cells (MSCs) Is Related to Unique Methylation Pattern in the Proximal Promoter as well as Gene Body DNA
Authors: Yen B. Linju , Hwa Hsiao-Lin , Hsu Pei-Ju , Chen Pei-Min , Wang Li-Tzu , Jiang Shih-Sheng , Liu Ko-Jiunn , Sytwu Huey-Kang , Yen Men-Luh
Published: July 2020

Multipotent human mesenchymal stem cells (MSCs) harbor clinically relevant immunomodulation, and HLA-G, a non-classical MHC class I molecule with highly restricted tissue expression, is one important molecule involved in these processes. Understanding of the natural regulatory mechanisms involved in expression of this elusive molecule has been difficult, with near exclusive reliance on cancer cell lines. We therefore studied the transcriptional control of HLA-G in primary isolated human bone marrow- (BM), human embryonic stem cell-derived (hE-), as well as placenta-derived MSCs (P-MSCs), and found that all 3 types of MSCs express 3 of the 7 HLA-G isoforms at the gene level; however, fibroblasts did not express HLA-G. Protein validation using BM- and P-MSCs demonstrated expression of 2 isoforms including a larger HLA-G-like protein. Interferon-γ (IFN-γ) stimulation upregulated both gene and protein expression in MSCs but not the constitutively expressing JEG-3 cell line. Most interestingly in human MSCs and placental tissue, hypomethylation of CpG islands not only occurs on the HLA-G proximal promoter but also on the gene body as well, a pattern not seen in either of the 2 commonly used choriocarcinoma cell lines which may contribute to the unique HLA-G expression patterns and IFN-γ-responsiveness in MSCs. Our study implicates the importance of using normal cells and tissues for physiologic understanding of tissue-specific transcriptional regulation, and highlight the utility of human MSCs in unraveling the transcriptional regulation of HLA-G for better therapeutic application.
Vesicular Transport of Encapsulated microRNA between Glial and Neuronal Cells
Authors: Lukiw Walter J. , Pogue Aileen I.
Published: July 2020

Exosomes (EXs) and extracellular microvesicles (EMVs) represent a diverse assortment of plasma membrane-derived nanovesicles, 30–1000 nm in diameter, released by all cell lineages of the central nervous system (CNS). They are examples of a very active and dynamic form of extracellular communication and the conveyance of biological information transfer essential to maintain homeostatic neurological functions and contain complex molecular cargoes representative of the cytoplasm of their cells of origin. These molecular cargoes include various mixtures of proteins, lipids, proteolipids, cytokines, chemokines, carbohydrates, microRNAs (miRNA) and messenger RNAs (mRNA) and other components, including end-stage neurotoxic and pathogenic metabolic products, such as amyloid beta (Aβ) peptides. Brain microglia, for example, respond to both acute CNS injuries and degenerative diseases with complex reactions via the induction of a pro-inflammatory phenotype, and secrete EXs and EMVs enriched in selective pathogenic microRNAs (miRNAs) such as miRNA-34a, miRNA-125b, miRNA-146a, miRNA-155, and others that are known to promote neuro-inflammation, induce complement activation, disrupt innate–immune signaling and deregulate the expression of neuron-specific phosphoproteins involved in neurotropism and synaptic signaling. This communication will review our current understanding of the trafficking of miRNA-containing EXs and EMVs from astrocytes and “activated pro-inflammatory” microglia to target neurons in neurodegenerative diseases with an emphasis on Alzheimer’s disease wherever possible.
Lipid Biomarkers as Predictors of Diastolic Dysfunction in Diabetes with Poor Glycemic Control
Authors: Khedr Dina , Hafez Mona , Lumpuy-Castillo Jairo , Emam Soha , Abdel-Massih Antoine , Elmougy Fatma , Elkaffas Rasha , Mahillo-Fernández Ignacio , Lorenzo Oscar , Musa Noha
Published: July 2020

Uncontrolled type-1 diabetes (T1DM) can lead to dyslipidaemia and albuminuria, which may promote cardiovascular injuries. However, some lipidemic factors could be useful in predicting cardiac dysfunction. Seventy-eight adolescents under insulin treatment due to a 6-year history of T1DM and were retrospectively examined. Glycemia, lipidemia, and albuminuria were measured in addition to development of cardiovascular abnormalities Both girls and boys showed higher HbA1c and fasting blood glucose and 27.1% females and 33.3% males exhibited microalbuminuria though their plasma levels of total cholesterol (TC), triglycerides (TG), and low-density lipoproteins (LDL) and high-density lipoproteins (HDL lipoproteins were in the normal range. They exhibited a preserved systolic function, but 50% of females and 66.6% of males had developed diastolic failures. Interestingly, girls with diastolic dysfunction showed significantly lower concentrations of HDL and higher TC/HDL and TG/HDL ratios. In fact, low HDL levels (OR 0.93; 95% CI 0.88–0.99; p = 0.029) and high TC/HDL (OR 2.55; 95% CI 1.9–5.45; p = 0.016) and TG/HDL (OR 2.74; 95% CI 1.12–6.71; p = 0.028) ratios associated with the development of diastolic complications. The cut-off values for HDL, TC/HDL, and TG/HDL were 49 mg/dL, 3.0 and 1.85, respectively. HDL and TC/HDL and TG/HDL ratios may be useful for predicting diastolic dysfunction in girls with uncontrolled T1DM.
A Novel Synthetic Compound (E)-5-((4-oxo-4H-chromen-3-yl)methyleneamino)-1-phenyl-1H-pyrazole-4-carbonitrile Inhibits TNFα-Induced MMP9 Expression via EGR-1 Downregulation in MDA-MB-231 Human Breast Cancer Cells
Authors: Jeong Munki , Jung Euitaek , Lee Young Han , Seo Jeong Kon , Ahn Seunghyun , Koh Dongsoo , Lim Yoongho , Shin Soon Young
Published: July 2020

Breast cancer is a common malignancy among women worldwide. Gelatinases such as matrix metallopeptidase 2 (MMP2) and MMP9 play crucial roles in cancer cell migration, invasion, and metastasis. To develop a novel platform compound, we synthesized a flavonoid derivative, (E)-5-((4-oxo-4H-chromen-3-yl)methyleneamino)-1-phenyl-1H-pyrazole-4-carbonitrile (named DK4023) and characterized its inhibitory effects on the motility and MMP2 and MMP9 expression of highly metastatic MDA-MB-231 breast cancer cells. We found that DK4023 inhibited tumor necrosis factor alpha (TNFα)-induced motility and F-actin formation of MDA-MB-231 cells. DK4023 also suppressed the TNFα-induced mRNA expression of MMP9 through the downregulation of the TNFα-extracellular signal-regulated kinase (ERK)/early growth response 1 (EGR-1) signaling axis. These results suggest that DK4023 could serve as a potential platform compound for the development of novel chemopreventive/chemotherapeutic agents against invasive breast cancer.
Loss of miR-145-5p Causes Ceruloplasmin Interference with PHD-Iron Axis and HIF-2α Stabilization in Lung Adenocarcinoma-Mediated Angiogenesis
Authors: Tsai Ying-Ming , Wu Kuan-Li , Chang Yung-Yun , Chang Wei-An , Huang Yung-Chi , Jian Shu-Fang , Tsai Pei-Hsun , Lin Yi-Shiuan , Chong Inn-Wen , Hung Jen-Yu , Hsu Ya-Ling
Published: July 2020

For decades, lung cancer has been the leading cause of cancer-related death worldwide. Hypoxia-inducible factors (HIFs) play critical roles in mediating lung cancer development and metastasis. The present study aims to clarify how HIF’s over-activation affects lung cancer angiogenesis not only in a normoxic condition, but also a hypoxic niche. Our study shows that human lung cancer exhibits elevated levels of ceruloplasmin (CP), which has a negative impact on the prognosis of patients. CP affects the cellular Fe2+ level, which inactivates prolyl hydroxylase (PHD) 1 and 2, resulting in HIF-2α enhancement. Increased HIF-2α leads to vascular endothelial growth factor-A (VEGF-A) secretion and angiogenesis. The expression of CP is under the epigenetic control of miR-145-5p. Restoration of miR-145-5p by miRNA mimics transfection decreases CP expression, increases Fe2+ and PHD1/2 levels and HIF hydroxylation while reduced HIF-2α levels resulting in the inhibition of tumor angiogenesis. In contrast, inhibition of miR-145-5p by miRNA inhibitors increases the expression of CP and VEGF-A in lung cancer cells. Significantly, miR-145-5p expression is lost in the tumor samples of lung cancer patients, and low miR-145-5p expression is strongly correlated with a shorter overall survival time. In conclusion, the current study reveals the clinical importance and prognostic value of miR-145-5p and CP. It identifies a unique mechanism of HIF-2α over-activation, which is mediated by iron imbalance of the iron-PHD coupling that modulates tumor angiogenesis.
Molecular Basis of the Pathogenic Mechanism Induced by the m.9191T>C Mutation in Mitochondrial ATP6 Gene
Authors: Su Xin , Dautant Alain , Godard François , Bouhier Marine , Zoladek Teresa , Kucharczyk Roza , di Rago Jean-Paul , Tribouillard-Tanvier Déborah
Published: July 2020

Probing the pathogenicity and functional consequences of mitochondrial DNA (mtDNA) mutations from patient’s cells and tissues is difficult due to genetic heteroplasmy (co-existence of wild type and mutated mtDNA in cells), occurrence of numerous mtDNA polymorphisms, and absence of methods for genetically transforming human mitochondria. Owing to its good fermenting capacity that enables survival to loss-of-function mtDNA mutations, its amenability to mitochondrial genome manipulation, and lack of heteroplasmy, Saccharomyces cerevisiae is an excellent model for studying and resolving the molecular bases of human diseases linked to mtDNA in a controlled genetic background. Using this model, we previously showed that a pathogenic mutation in mitochondrial ATP6 gene (m.9191T>C), that converts a highly conserved leucine residue into proline in human ATP synthase subunit a (aL222P), severely compromises the assembly of yeast ATP synthase and reduces by 90% the rate of mitochondrial ATP synthesis. Herein, we report the isolation of intragenic suppressors of this mutation. In light of recently described high resolution structures of ATP synthase, the results indicate that the m.9191T>C mutation disrupts a four α-helix bundle in subunit a and that the leucine residue it targets indirectly optimizes proton conduction through the membrane domain of ATP synthase.
Combination of NKT14m and Low Dose IL-12 Promotes Invariant Natural Killer T Cell IFN-γ Production and Tumor Control
Authors: Guan Peng , Schaub Robert , Nichols Kim E. , Das Rupali
Published: July 2020

Invariant natural killer T (iNKT) cells are innate-like T lymphocytes characterized by the expression of an invariant T cell receptor (iTCR) that recognizes glycolipid antigens presented by the MHC I-like CD1d molecule. Following antigenic stimulation, iNKT cells rapidly produce large amounts of cytokines that can trans-activate dendritic cells (DC) and promote the anti-tumor functions of cytotoxic lymphocytes, such as natural killer (NK) and CD8 T cells. Additionally, iNKT cells can mediate robust and direct cytotoxicity against CD1d+ tumor targets. However, many tumors down-regulate CD1d and evade iNKT cell attack. To circumvent this critical barrier to iNKT cell anti-tumor activity, a novel monoclonal antibody (mAb), NKT14 has been recently developed. This agonistic antibody binds directly and specifically to the iTCR of murine iNKT cells. In the current study, we demonstrate that NKT14m mediates robust activation, cytokine production and degranulation of murine iNKT cells, in vitro. Consistently, NKT14m also promoted iNKT cell activation and immunomodulatory functions, in vivo. Finally, administration of NKT14m with low dose interleukin (IL)-12 further augmented iNKT cell IFN-γ production in vivo, and this combination conferred superior suppression of tumor cell growth compared to NKT14m or IL-12 alone. Together, these data demonstrate that a combination treatment consisting of low dose IL-12 and iTCR-specific mAb may be an attractive alternative to activate iNKT cell anti-tumor functions.
Molecular Mechanisms, Epidemiology, and Clinical Importance of β-Lactam Resistance in Enterobacteriaceae
Authors: De Angelis Giulia , Del Giacomo Paola , Posteraro Brunella , Sanguinetti Maurizio , Tumbarello Mario
Published: July 2020

Despite being members of gut microbiota, Enterobacteriaceae are associated with many severe infections such as bloodstream infections. The β-lactam drugs have been the cornerstone of antibiotic therapy for such infections. However, the overuse of these antibiotics has contributed to select β-lactam-resistant Enterobacteriaceae isolates, so that β-lactam resistance is nowadays a major concern worldwide. The production of enzymes that inactivate β-lactams, mainly extended-spectrum β-lactamases and carbapenemases, can confer multidrug resistance patterns that seriously compromise therapeutic options. Further, β-lactam resistance may result in increases in the drug toxicity, mortality, and healthcare costs associated with Enterobacteriaceae infections. Here, we summarize the updated evidence about the molecular mechanisms and epidemiology of β-lactamase-mediated β-lactam resistance in Enterobacteriaceae, and their potential impact on clinical outcomes of β-lactam-resistant Enterobacteriaceae infections.
Carbonic Anhydrase VI in Skin Wound Healing Study on Car6 Knockout Mice
Authors: Pemmari Toini , Laakso Jaakko , Patrikainen Maarit S. , Parkkila Seppo , Järvinen Tero A. H.
Published: July 2020

Carbonic anhydrases (CAs) contribute to tumor cell migration by generating an acidic environment through the conversion of carbon dioxide to bicarbonate and a proton. CA VI is secreted to milk and saliva, and it could contribute to wound closure, as a potential trophic factor, in animals that typically lick their wounds. Our aim was to investigate whether human CA VI improves skin-wound healing in full-thickness skin-wound models. The effect was studied in Car6 −/− knockout mice and wild type littermates. Half of both mice strains were given topically administered, milk-derived CA VI after wounding and eight hours later. The amount of topically given CA VI exceeded the predicted amount of natural saliva-delivered CA VI. The healing was followed for seven days and studied from photographs and histological sections. Our results showed no significant differences between the treatment groups in wound closure, re-epithelization, or granulation tissue formation, nor did the Car6 genotype affect the healing. Our results demonstrate that CA VI does not play a major role in skin-wound healing and also suggest that saliva-derived CA VI is not responsible for the licking-associated improved wound healing in animals.
Transcriptomic Profiling of Young Cotyledons Response to Chilling Stress in Two Contrasting Cotton (Gossypium hirsutum L.) Genotypes at the Seedling Stage
Authors: Cheng Gongmin , Zhang Longyan , Wang Hantao , Lu Jianhua , Wei Hengling , Yu Shuxun
Published: July 2020

Young cotyledons of cotton seedlings are most susceptible to chilling stress. To gain insight into the potential mechanism of cold tolerance of young cotton cotyledons, we conducted physiological and comparative transcriptome analysis of two varieties with contrasting phenotypes. The evaluation of chilling injury of young cotyledons among 74 cotton varieties revealed that H559 was the most tolerant and YM21 was the most sensitive. The physiological analysis found that the ROS scavenging ability was lower, and cell membrane damage was more severe in the cotyledons of YM21 than that of H559 under chilling stress. RNA-seq analysis identified a total of 44,998 expressed genes and 19,982 differentially expressed genes (DEGs) in young cotyledons of the two varieties under chilling stress. Weighted gene coexpression network analysis (WGCNA) of all DEGs revealed four significant modules with close correlation with specific samples. The GO-term enrichment analysis found that lots of genes in H559-specific modules were involved in plant resistance to abiotic stress. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that pathways such as plant hormone signal transduction, MAPK signaling, and plant–pathogen interaction were related to chilling stress response. A total of 574 transcription factors and 936 hub genes in these modules were identified. Twenty hub genes were selected for qRT-PCR verification, revealing the reliability and accuracy of transcriptome data. These findings will lay a foundation for future research on the molecular mechanism of cold tolerance in cotyledons of cotton.
Modulated Electro-Hyperthermia Resolves Radioresistance of Panc1 Pancreas Adenocarcinoma and Promotes DNA Damage and Apoptosis In Vitro
Authors: Forika Gertrud , Balogh Andrea , Vancsik Tamas , Zalatnai Attila , Petovari Gabor , Benyo Zoltan , Krenacs Tibor
Published: July 2020

The poor outcome of pancreas ductal adenocarcinomas (PDAC) is frequently linked to therapy resistance. Modulated electro-hyperthermia (mEHT) generated by 13.56 MHz capacitive radiofrequency can induce direct tumor damage and promote chemo- and radiotherapy. Here, we tested the effect of mEHT either alone or in combination with radiotherapy using an in vivo model of Panc1, a KRAS and TP53 mutant, radioresistant PDAC cell line. A single mEHT shot of 60 min induced ~50% loss of viable cells and morphological signs of apoptosis including chromatin condensation, nuclear shrinkage and apoptotic bodies. Most mEHT treatment related effects exceeded those of radiotherapy, and these were further amplified after combining the two modalities. Treatment related apoptosis was confirmed by a significantly elevated number of annexin V single-positive and cleaved/activated caspase-3 positive tumor cells, as well as sub-G1-phase tumor cell fractions. mEHT and mEHT+radioterapy caused the moderate accumulation of γH2AX positive nuclear foci, indicating DNA double-strand breaks and upregulation of the cyclin dependent kinase inhibitor p21waf1 besides the downregulation of Akt signaling. A clonogenic assay revealed that both mono- and combined treatments affected the tumor progenitor/stem cell populations too. In conclusion, mEHT treatment can contribute to tumor growth inhibition and apoptosis induction and resolve radioresistance of Panc1 PDAC cells.
PEG-8 Laurate Fermentation of Staphylococcus epidermidis Reduces the Required Dose of Clindamycin Against Cutibacterium acnes
Authors: Marito Shinta , Keshari Sunita , Huang Chun-Ming
Published: July 2020

The probiotic activity of skin Staphylococcus epidermidis (S. epidermidis) bacteria can elicit diverse biological functions via the fermentation of various carbon sources. Here, we found that polyethylene glycol (PEG)-8 Laurate, a carbon-rich molecule, can selectively induce the fermentation of S. epidermidis, not Cutibacterium acnes (C. acnes), a bacterium associated with acne vulgaris. The PEG-8 Laurate fermentation of S. epidermidis remarkably diminished the growth of C. acnes and the C. acnes-induced production of pro-inflammatory macrophage-inflammatory protein 2 (MIP-2) cytokines in mice. Fermentation media enhanced the anti-C. acnes activity of a low dose (0.1%) clindamycin, a prescription antibiotic commonly used to treat acne vulgaris, in terms of the suppression of C. acnes colonization and MIP-2 production. Furthermore, PEG-8 Laurate fermentation of S. epidermidis boosted the activity of 0.1% clindamycin to reduce the sizes of C. acnes colonies. Our results demonstrated, for the first time, that the PEG-8 Laurate fermentation of S. epidermidis displayed the adjuvant effect on promoting the efficacy of low-dose clindamycin against C. acnes. Targeting C. acnes by lowering the required doses of antibiotics may avoid the risk of creating drug-resistant C. acnes and maintain the bacterial homeostasis in the skin microbiome, leading to a novel modality for the antibiotic treatment of acne vulgaris.
An Integrative Analysis of Transcriptome, Proteome and Hormones Reveals Key Differentially Expressed Genes and Metabolic Pathways Involved in Flower Development in Loquat
Authors: Jing Danlong , Chen Weiwei , Hu Ruoqian , Zhang Yuchen , Xia Yan , Wang Shuming , He Qiao , Guo Qigao , Liang Guolu
Published: July 2020

Flower development is a vital developmental process in the life cycle of woody perennials, especially fruit trees. Herein, we used transcriptomic, proteomic, and hormone analyses to investigate the key candidate genes/proteins in loquat (Eriobotrya japonica) at the stages of flower bud differentiation (FBD), floral bud elongation (FBE), and floral anthesis (FA). Comparative transcriptome analysis showed that differentially expressed genes (DEGs) were mainly enriched in metabolic pathways of hormone signal transduction and starch and sucrose metabolism. Importantly, the DEGs of hormone signal transduction were significantly involved in the signaling pathways of auxin, gibberellins (GAs), cytokinin, ethylene, abscisic acid (ABA), jasmonic acid, and salicylic acid. Meanwhile, key floral integrator genes FLOWERING LOCUS T (FT) and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1) and floral meristem identity genes SQUAMOSA PROMOTER BINDING LIKE (SPL), LEAFY (LFY), APETALA1 (AP1), and AP2 were significantly upregulated at the FBD stage. However, key floral organ identity genes AGAMOUS (AG), AP3, and PISTILLATA (PI) were significantly upregulated at the stages of FBE and FA. Furthermore, transcription factors (TFs) such as bHLH (basic helix-loop-helix), NAC (no apical meristem (NAM), Arabidopsis transcription activation factor (ATAF1/2) and cup-shaped cotyledon (CUC2)), MYB_related (myeloblastosis_related), ERF (ethylene response factor), and C2H2 (cysteine-2/histidine-2) were also significantly differentially expressed. Accordingly, comparative proteomic analysis of differentially accumulated proteins (DAPs) and combined enrichment of DEGs and DAPs showed that starch and sucrose metabolism was also significantly enriched. Concentrations of GA3 and zeatin were high before the FA stage, but ABA concentration remained high at the FA stage. Our results provide abundant sequence resources for clarifying the underlying mechanisms of the flower development in loquat.
FOXC2 Disease Mutations Identified in Lymphedema Distichiasis Patients Impair Transcriptional Activity and Cell Proliferation
Authors: Tavian Daniela , Missaglia Sara , Michelini Sandro , Maltese Paolo Enrico , Manara Elena , Mordente Alvaro , Bertelli Matteo
Published: July 2020

FOXC2 is a member of the human forkhead-box gene family and encodes a regulatory transcription factor. Mutations in FOXC2 have been associated with lymphedema distichiasis (LD), an autosomal dominant disorder that primarily affects the limbs. Most patients also show extra eyelashes, a condition known as distichiasis. We previously reported genetic and clinical findings in six unrelated families with LD. Half the patients showed missense mutations, two carried frameshift mutations and a stop mutation was identified in a last patient. Here we analyzed the subcellular localization and transactivation activity of the mutant proteins, showing that all but one (p.Y109*) localized to the nucleus. A significant reduction of transactivation activity was observed in four mutants (p.L80F, p.H199Pfs*264, p.I213Tfs*18, p.Y109*) compared with wild type FOXC2 protein, while only a partial loss of function was associated with p.V228M. The mutant p.I213V showed a very slight increase of transactivation activity. Finally, immunofluorescence analysis revealed that some mutants were sequestered into nuclear aggregates and caused a reduction of cell viability. This study offers new insights into the effect of FOXC2 mutations on protein function and shows the involvement of aberrant aggregation of FOXC2 proteins in cell death.
Controlling the Kinetics of an Enzymatic Reaction through Enzyme or Substrate Confinement into Lipid Mesophases with Tunable Structural Parameters
Authors: Mendozza Marco , Balestri Arianna , Montis Costanza , Berti Debora
Published: July 2020

Lipid liquid crystalline mesophases, resulting from the self-assembly of polymorphic lipids in water, have been widely explored as biocompatible drug delivery systems. In this respect, non-lamellar structures are particularly attractive: they are characterized by complex 3D architectures, with the coexistence of hydrophobic and hydrophilic regions that can conveniently host drugs of different polarities. The fine tunability of the structural parameters is nontrivial, but of paramount relevance, in order to control the diffusive properties of encapsulated active principles and, ultimately, their pharmacokinetics and release. In this work, we investigate the reaction kinetics of p-nitrophenyl phosphate conversion into p-nitrophenol, catalysed by the enzyme Alkaline Phosphatase, upon alternative confinement of the substrate and of the enzyme into liquid crystalline mesophases of phytantriol/H2O containing variable amounts of an additive, sucrose stearate, able to swell the mesophase. A structural investigation through Small-Angle X-ray Scattering, revealed the possibility to finely control the structure/size of the mesophases with the amount of the included additive. A UV–vis spectroscopy study highlighted that the enzymatic reaction kinetics could be controlled by tuning the structural parameters of the mesophase, opening new perspectives for the exploitation of non-lamellar mesophases for confinement and controlled release of therapeutics.
Carotid Body and Metabolic Syndrome: Mechanisms and Potential Therapeutic Targets
Authors: Kim Lenise J. , Polotsky Vsevolod Y.
Published: July 2020

The carotid body (CB) is responsible for the peripheral chemoreflex by sensing blood gases and pH. The CB also appears to act as a peripheral sensor of metabolites and hormones, regulating the metabolism. CB malfunction induces aberrant chemosensory responses that culminate in the tonic overactivation of the sympathetic nervous system. The sympatho-excitation evoked by CB may contribute to the pathogenesis of metabolic syndrome, inducing systemic hypertension, insulin resistance and sleep-disordered breathing. Several molecular pathways are involved in the modulation of CB activity, and their pharmacological manipulation may lead to overall benefits for cardiometabolic diseases. In this review, we will discuss the role of the CB in the regulation of metabolism and in the pathogenesis of the metabolic dysfunction induced by CB overactivity. We will also explore the potential pharmacological targets in the CB for the treatment of metabolic syndrome.
The T2 Toxin Produced by Fusarium spp. Impacts Porcine Duodenal Nitric Oxide Synthase (nNOS)-Positive Nervous Structures—The Preliminary Study
Authors: Rychlik Andrzej , Gonkowski Slawomir , Kaczmar Ewa , Obremski Kazimierz , Calka Jaroslaw , Makowska Krystyna
Published: July 2020

T2 toxin synthetized by Fusarium spp. negatively affects various internal organs and systems, including the digestive tract and the immune, endocrine, and nervous systems. However, knowledge about the effects of T2 on the enteric nervous system (ENS) is still incomplete. Therefore, during the present experiment, the influence of T2 toxin with a dose of 12 µg/kg body weight (b.w.)/per day on the number of enteric nervous structures immunoreactive to neuronal isoform nitric oxide synthase (nNOS—used here as a marker of nitrergic neurons) in the porcine duodenum was studied using the double immunofluorescence method. Under physiological conditions, nNOS-positive neurons amounted to 38.28 ± 1.147%, 38.39 ± 1.244%, and 35.34 ± 1.151 of all enteric neurons in the myenteric (MP), outer submucous (OSP), and inner submucous (ISP) plexuses, respectively. After administration of T2 toxin, an increase in the number of these neurons was observed in all types of the enteric plexuses and nNOS-positive cells reached 46.20 ± 1.453% in the MP, 45.39 ± 0.488% in the OSP, and 44.07 ± 0.308% in the ISP. However, in the present study, the influence of T2 toxin on the intramucosal and intramuscular nNOS-positive nerves was not observed. The results obtained in the present study indicate that even low doses of T2 toxin are not neutral for living organisms because they may change the neurochemical characterization of the enteric neurons.
Discrimination of CpG Methylation Status and Nucleotide Differences in Tissue Specimen DNA by Oligoribonucleotide Interference-PCR
Authors: Shimizu Takeshi , Fujita Toshitsugu , Fukushi Sakie , Horino Yuri , Fujii Hodaka
Published: July 2020

Oligoribonucleotide (ORN) interference-PCR (ORNi-PCR) is a method in which PCR amplification of a target sequence is inhibited in a sequence-specific manner by the hybridization of an ORN with the target sequence. Previously, we reported that ORNi-PCR could detect nucleotide mutations in DNA purified from cultured cancer cell lines or genome-edited cells. In this study, we investigated whether ORNi-PCR can discriminate nucleotide differences and CpG methylation status in damaged DNA, such as tissue specimen DNA and bisulfite-treated DNA. First, we showed that ORNi-PCR could discriminate nucleotide differences in DNA extracted from acetone-fixed paraffin-embedded rat liver specimens or formalin-fixed paraffin-embedded human specimens. Rat whole blood specimens were compatible with ORNi-PCR for the same purpose. Next, we showed that ORNi-PCR could discriminate CpG methylation status in bisulfite-treated DNA. These results demonstrate that ORNi-PCR can discriminate nucleotide differences and CpG methylation status in multiple types of DNA samples. Thus, ORNi-PCR is potentially useful in a wide range of fields, including molecular biology and medical diagnosis.