bluuebunny/crossref_metadata_2025 huggingface dataset:
Synthesizing Observations and Theory to Understand Galactic Magnetic Fields: Progress and Challenges
Authors: Beck Rainer , Chamandy Luke , Elson Ed , Blackman Eric G.
Published: December 2019

Constraining dynamo theories of magnetic field origin by observation is indispensable but challenging, in part because the basic quantities measured by observers and predicted by modelers are different. We clarify these differences and sketch out ways to bridge the divide. Based on archival and previously unpublished data, we then compile various important properties of galactic magnetic fields for nearby spiral galaxies. We consistently compute strengths of total, ordered, and regular fields, pitch angles of ordered and regular fields, and we summarize the present knowledge on azimuthal modes, field parities, and the properties of non-axisymmetric spiral features called magnetic arms. We review related aspects of dynamo theory, with a focus on mean-field models and their predictions for large-scale magnetic fields in galactic discs and halos. Furthermore, we measure the velocity dispersion of H i gas in arm and inter-arm regions in three galaxies, M 51, M 74, and NGC 6946, since spiral modulation of the root-mean-square turbulent speed has been proposed as a driver of non-axisymmetry in large-scale dynamos. We find no evidence for such a modulation and place upper limits on its strength, helping to narrow down the list of mechanisms to explain magnetic arms. Successes and remaining challenges of dynamo models with respect to explaining observations are briefly summarized, and possible strategies are suggested. With new instruments like the Square Kilometre Array (SKA), large data sets of magnetic and non-magnetic properties from thousands of galaxies will become available, to be compared with theory.
Functionally Annotating Regulatory Elements in the Equine Genome Using Histone Mark ChIP-Seq
Authors: Kingsley N. B. , Kern Colin , Creppe Catherine , Hales Erin N. , Zhou Huaijun , Kalbfleisch T. S. , MacLeod James N. , Petersen Jessica L. , Finno Carrie J. , Bellone Rebecca R.
Published: December 2019

One of the primary aims of the Functional Annotation of ANimal Genomes (FAANG) initiative is to characterize tissue-specific regulation within animal genomes. To this end, we used chromatin immunoprecipitation followed by sequencing (ChIP-Seq) to map four histone modifications (H3K4me1, H3K4me3, H3K27ac, and H3K27me3) in eight prioritized tissues collected as part of the FAANG equine biobank from two thoroughbred mares. Data were generated according to optimized experimental parameters developed during quality control testing. To ensure that we obtained sufficient ChIP and successful peak-calling, data and peak-calls were assessed using six quality metrics, replicate comparisons, and site-specific evaluations. Tissue specificity was explored by identifying binding motifs within unique active regions, and motifs were further characterized by gene ontology (GO) and protein–protein interaction analyses. The histone marks identified in this study represent some of the first resources for tissue-specific regulation within the equine genome. As such, these publicly available annotation data can be used to advance equine studies investigating health, performance, reproduction, and other traits of economic interest in the horse.
Functional Genomic Identification of Cadmium Resistance Genes from a High GC Clone Library by Coupling the Sanger and PacBio Sequencing Strategies
Authors: Chen Jinghao , Xing Chao , Zheng Xin , Li Xiaofang
Published: December 2019

Functional (meta) genomics allows the high-throughput identification of functional genes in a premise-free way. However, it is still difficult to perform Sanger sequencing for high GC DNA templates, which hinders the functional genomic exploration of a high GC genomic library. Here, we developed a procedure to resolve this problem by coupling the Sanger and PacBio sequencing strategies. Identification of cadmium (Cd) resistance genes from a small-insert high GC genomic library was performed to test the procedure. The library was generated from a high GC (75.35%) bacterial genome. Nineteen clones that conferred Cd resistance to Escherichia coli subject to Sanger sequencing directly. The positive clones were in parallel subject to in vivo amplification in host cells, from which recombinant plasmids were extracted and linearized by selected restriction endonucleases. PacBio sequencing was performed to obtain the full-length sequences. As the identities, partial sequences from Sanger sequencing were aligned to the full-length sequences from PacBio sequencing, which led to the identification of seven unique full-length sequences. The unique sequences were further aligned to the full genome sequence of the source strain. Functional screening showed that the identified positive clones were all able to improve Cd resistance of the host cells. The functional genomic procedure developed here couples the Sanger and PacBio sequencing methods and overcomes the difficulties in PCR approaches for high GC DNA. The procedure can be a promising option for the high-throughput sequencing of functional genomic libraries, and realize a cost-effective and time-efficient identification of the positive clones, particularly for high GC genetic materials.
Effect of Land Use Conversion on Surface Soil Heavy Metal Contamination in a Typical Karst Plateau Lakeshore Wetland of Southwest China
Authors: Sun Caili , Zhu Sixi , Zhao Bin , Li Wujiang , Gao Xiaoye , Wang Xiaodan
Published: December 2019

Land use conversion could directly or indirectly influence heavy metal geochemistry by changing soil properties. The aim of this study was to explore the effect of land use conversion on surface soil heavy metal contamination in the karst plateau lakeshore wetlands of Southwest China. Based on this, a total of 120 soil samples were collected from 30 sites from different types of land uses (farmlands, grasslands and woodlands) around a lake in Suohuangcang National Wetland Park in August 2017. Contents of As, Cd, Cu, Cr, Hg, Pb and Zn were analyzed, and soil heavy metal contamination was assessed in all three land use types. Results showed that land use transformation from farmland to grassland or woodland was not conducive to the release of soil heavy metal. Surface soil of all three land use types have been moderately polluted by As, Cr, Pb, and Zn, and grassland and woodland also had moderate Cd contamination. The pollution load index (PLI) results revealed low heavy metal contamination in grassland and woodland but no contamination in farmland. Although the integrated contamination in the studied region did not pose a serious potential ecological risk (RI < 150), it might affect human health through the water supply and food chain. Therefore, it is necessary to monitor and control As, Cd, Cr, Pb, and Zn concentrations of surface soil through controlling pollutants, improving waste treatment, as well as strengthening supervision and management in the vicinity of the Suohuangcang National Wetland Park.
Nutrition Education Practices of Health Teachers from Shanghai K-12 Schools: The Current Status, Barriers and Willingness to Teach
Authors: Li Fan , Yuan Yaqun , Xu Xinming , Chen Jingsi , Li Jiaxuan , He Gengsheng , Chen Bo
Published: December 2019

China is facing challenges in both undernutrition and overnutrition, resulting from unhealthy diets. Nutrition education early in life, especially in school settings, has been reported to be effective in addressing these challenges. However, little is known about how nutrition education is delivered in schools in China. This study aimed to investigate the current status of delivering nutrition education by health teachers in Shanghai and to determine the barriers and resources that influence the teachers’ practices and their willingness to teach nutrition. In 2016–2017, a cross-sectional questionnaire survey was conducted on 904 health teachers from 823 K-12 schools in Shanghai, China. There were 722 (79.9%) teachers that had the experience of teaching nutrition, but only 137 (19.0% of 722) spent ≥1 h teaching nutrition courses in each school year. Only 18.6% of the teachers had received a formal education in nutrition in college. About 88.5% of teachers expressed their willingness to teach nutrition in the future. The three major reasons for never teaching nutrition were categorized as: nutrition being taught by other teachers (39.5%), willing to teach but lack of knowledge (37.9%), and the subject not being required by school administrators (31.3%). Teachers who spent more time or were more willing to teach nutrition courses were those who were female, from private schools, had a better background in receiving nutrition education, and were more concerned about nutrition. Our data show that nutrition education is at a formative stage in Shanghai, China.
Predicting Bullying through Motivation and Teaching Styles in Physical Education
Authors: Montero-Carretero Carlos , Barbado David , Cervelló Eduardo
Published: December 2019

From the theoretical framework offered by the self-determination theory, the objective of the study was to test a predictor model of bullying behaviors based on the physical education teacher’s supportive style, the students’ satisfaction of basic psychological needs, and self-determined motivation. A total of 608 students of both sexes, between 11 and 15 years of age, from primary and secondary schools in the province of Alicante (Spain) voluntarily completed questionnaires to measure each of the variables under study. The design of the study was cross-sectional. The results showed that the autonomy supportive style positively predicted the satisfaction of basic psychological needs, which, in turn, positively predicted self-determined motivation towards physical education. The latter negatively predicted bullying perpetration and bullying victimization. The controlling style presented inverse relationships to those of the autonomy supportive style. These results are in line with the positions of the self-determination theory and underline the potential responsibility of physical education teachers in the struggle against bullying, and how, by supporting autonomy and avoiding a controlling style, they can help reduce bullying perpetration and victimization.
Fruit and Vegetable Purchases in Farmer’s Market Stands: Analysing Survey and Sales Data
Authors: Rebouillat Pauline , Bonin Sarah , Kestens Yan , Chaput Sarah , Drouin Louis , Mercille Geneviève
Published: December 2019

Farmers’ market implementation holds promise for increasing access to healthy foods. Although rarely measured, purchase data constitute an intermediate outcome between food environment and actual consumption. In a study conducted with two seasonal Fruits and Vegetables (FV) stands in a disadvantaged area of Montréal (Canada), we analysed how accessibility, perception, and mobility-related factors were associated with FV purchase. This analysis uses a novel measure of FV purchasing practices based on sales data obtained from a mobile application. A 2016 survey collected information on markets’ physical access, perceived access to FV in the neighbourhood, usual FV consumption and purchases. Multivariate models were used to analyse three purchasing practice indicators: number of FV portions, FV variety and expenditures. Average shoppers purchased 12 FV portions of three distinct varieties and spent 5$. Shoppers stopping at the market on their usual travel route spent less (p = 0.11), bought fewer portions (p = 0.03) and a lesser FV variety (p < 0.01). FV stands may complement FV dietary intake. Individuals for whom the market is on their usual travel route might make more frequent visits and, therefore, smaller purchases. The novel data collection method allowed analysis of multiple purchase variables, is precise and easy to apply at unconventional points of sales and could be transposed elsewhere.
Youth Engaged Participatory Air Monitoring: A ‘Day in the Life’ in Urban Environmental Justice Communities
Authors: Johnston Jill E. , Juarez Zully , Navarro Sandy , Hernandez Ashley , Gutschow Wendy
Published: December 2019

Air pollution in Southern California does not impact all communities equally; communities of color are disproportionately burdened by poor air quality and more likely to live near industrial facilities and freeways. Government regulatory monitors do not have the spatial resolution to provide air quality information at the neighborhood or personal scale. We describe the A Day in the Life program, an approach to participatory air monitoring that engages youth in collecting data that they can then analyze and use to take action. Academics partnered with Los Angeles-based youth environmental justice organizations to combine personal air monitoring, participatory science, and digital storytelling to build capacity to address local air quality issues. Eighteen youth participants from four different neighborhoods wore portable personal PM2.5 (fine particles <2.5 µm in diameter) monitors for a day in each of their respective communities, documenting and mapping their exposure to PM2.5 during their daily routine. Air monitoring was coupled with photography and videos to document what they experienced over the course of their day. The PM2.5 exposure during the day for participants averaged 10.7 µg/m3, although the range stretched from <1 to 180 µg/m3. One-third of all measurements were taken <300 m from a freeway. Overall, we demonstrate a method to increase local youth-centered understanding of personal exposures, pollution sources, and vulnerability to air quality.
Evidence of Adaptation to Increasing Temperatures
Authors: Weitensfelder Lisbeth , Moshammer Hanns
Published: December 2019

In times of rising temperatures, the question arises on how the human body adapts. When assumed that changing climate leads to adaptation, time series analysis should reveal a shift in optimal temperatures. The city of Vienna is especially affected by climate change due to its location in the Alpine Range in Middle Europe. Based on mortality data, we calculated shifts in optimal temperature for a time period of 49 years in Vienna with Poisson regression models. Results show a shift in optimal temperature, with optimal temperature increasing more than average temperature. Hence, results clearly show an adaptation process, with more adaptation to warmer than colder temperatures. Nevertheless, some age groups remain more vulnerable than others and less able to adapt. Further research focusing on vulnerable groups should be encouraged.
Childcare Center Characteristics Moderate the Effects of a Physical Activity Intervention
Authors: Saunders Ruth P. , Dowda Marsha , Pfeiffer Karin A. , Brown William H. , Pate Russell R.
Published: December 2019

Center-based early childhood education and care (ECEC) programs are well-positioned to create positive impacts on the health and development of large numbers of young children by promoting physical activity using evidence-based programs. Studies testing physical activity programs for young children should examine the circumstances under which programs are most effective by assessing the role of contextual factors on program outcomes. The purpose of this study was to examine the moderating effects of baseline ECEC center characteristics on the relationship between the Study of Health and Activity in Preschool Environments (SHAPES) intervention and moderate-to-vigorous physical activity (MVPA). MVPA was assessed via accelerometry; center characteristics, practices, and social and physical environments were assessed by director interview and observation; and center quality was assessed using the Early Childhood Environment Rating Scale-Revised Edition. Mixed-model analyses of covariance (ANCOVAs) examined intervention effects on MVPA during the school day; interactions between baseline center variables and group assignment (intervention vs. control) tested for moderation. Two center instructional practices, two social environment characteristics, and one physical environment characteristic at baseline moderated the effects of SHAPES on MVPA outcomes. Assessing baseline practices and center characteristics may aid efforts to match centers with interventions likely to increase physical activity as well as suggest additional intervention strategies to test.
Mortality in Via Ferrata Emergencies in Austria from 2008 to 2018
Authors: Ströhle Mathias , Haselbacher Matthias , Rugg Christopher , Walpoth Alex , Konetschny Ricarda , Paal Peter , Mair Peter
Published: December 2019

Although the European Alps now have more than 1000 via ferratas, limited data exist on the actual incidence of fatal events in via ferratas and their causes. This retrospective study analysed data from a registry maintained by the Austrian Alpine Safety Board (n = 161,855, per 11 September 2019). Over a 10-year period from 1 November 2008 to 31 October 2018, all persons involved in a via ferrata-related emergency were included (n = 1684), of which 64% were male. Most emergencies were caused by blockage due to exhaustion and/or misjudgement of the climber’s own abilities. Consequently, more than half of all victims were evacuated uninjured. Only 62 (3.7%) via ferrata-related deaths occurred. Falling while climbing unsecured was the most common cause of death, and males had a 2.5-fold higher risk of dying in a via ferrata accident. The mortality rate was highest in technically easy-to-climb sections (Grade A, 13.2%/B, 4.9%), whereas the need to be rescued uninjured was highest in difficult routes (Grade D, 59.9%/E, 62.7%). Although accidents in via ferratas are common and require significant rescue resources, fatal accidents are rare. The correct use of appropriate equipment in technically easy-to-climb routes can prevent the majority of these fatalities.
Bisphenols as a Legacy Pollutant, and Their Effects on Organ Vulnerability
Authors: Kim Jong-Joo , Kumar Surendra , Kumar Vinay , Lee Yun-Mi , Kim You-Sam , Kumar Vijay
Published: December 2019

Bisphenols are widely used in the synthesis of polycarbonate plastics, epoxy resins, and thermal paper, which are used in manufacturing items of daily use. Packaged foods and drinks are the main sources of exposure to bisphenols. These chemicals affect humans and animals by disrupting the estrogen, androgen, progesterone, thyroid, and aryl hydrocarbon receptor functions. Bisphenols exert numerous harmful effects because of their interaction with receptors, reactive oxygen species (ROS) formation, lipid peroxidation, mitochondrial dysfunction, and cell signal alterations. Both cohort and case-control studies have determined an association between bisphenol exposure and increased risk of cardiovascular diseases, neurological disorders, reproductive abnormalities, obesity, and diabetes. Prenatal exposure to bisphenols results in developmental disorders in animals. These chemicals also affect the immune cells and play a significant role in initiating the inflammatory response. Exposure to bisphenols exhibit age, gender, and dose-dependent effects. Even at low concentrations, bisphenols exert toxicity, and hence deserve a critical assessment of their uses. Since bisphenols have a global influence on human health, the need to discover the underlying pathways involved in all disease conditions is essential. Furthermore, it is important to promote the use of alternatives for bisphenols, thereby restricting their uses.
Employment Experiences among Young Malaysian Adults with Learning Disabilities
Authors: Harun Dzalani , Che’ Din Normah , Mohd Rasdi Hanif Farhan , Shamsuddin Khadijah
Published: December 2019

The aim of this study was to describe the employment experiences of persons with learning disabilities (LDs) in developing countries, such as Malaysia. Factors associated with respondents’ employment were also determined. A cross-sectional survey was conducted among young adults with LD who left the special education programs in secondary schools in Kuala Lumpur and Selangor. Ninety young adults with LD, aged 18 to 25 years were interviewed face-to-face at an agreed upon convenient place on their working experiences after leaving secondary schools. A total of 13 respondents were excluded from the analysis because their intellectual quotient (IQ) score demonstrated a high possibility of intellectual disability with IQ estimation <70. Of the 77 young adults analyzed, 74.0% reported having work experience and 64.9% were working at the time of interview. Statistical analysis showed significant associations between individual, family, and community factors with respondents’ employment. Two factors made a unique statistically significant contribution to the model (gender, p = 0.043 and adult service: Financial support p = 0.012). This study suggests the current school-to-work transition program at secondary and post-secondary schools should be improved to better prepare young adults with LD with necessary skills relevant for the current job market so that they could improve their employability.
How Do Type 2 Diabetes Patients Value Urban Integrated Primary Care in China? Results of a Discrete Choice Experiment
Authors: Wang Xin , Song Kuimeng , Zhu Paiyi , Valentijn Pim , Huang Yixiang , Birch Stephen
Published: December 2019

Objectives: Fragmented healthcare in China cannot meet the needs of the growing number of type 2 diabetes patients. The World Health Organization proposed an integrated primary care approach to address the needs of patients with chronic conditions. This study aims to measure type 2 diabetes patients’ preferences for urban integrated primary care in China. Methods: A discrete choice experiment was designed to measure type 2 diabetes patient preferences for seven priority attributes of integrated care. A two-stage sampling survey of 307 type 2 diabetes mellitus (T2DM) patients in 16 community health stations was carried out. Interviews were conducted to explore the reasons underpinning the preferences. A logit regression model was used to estimate patients’ willingness to pay and to analyze the expected impact of potential policy changes. Results: Travel time to care providers and experience of care providers are the most valued attributes for respondents rather than out-of-pocket cost. Attention to personal situation, the attentiveness of care providers, and the friendliness and helpfulness of staff were all related to interpersonal communication between patients and health care providers. Accurate health information and multidisciplinary care were less important attributes. Conclusions: The study provides an insight into type 2 diabetes patients’ needs and preferences of integrated primary care. People-centered interventions, such as increasing coverage by family doctor and cultivating mutual continuous relationships appear to be key priorities of policy and practice in China.
The Association between Living Status Transitions, Behavior Changes and Family Relationship Improvement among Methadone Maintenance Treatment Participants in Guangdong, China
Authors: Liu Shaokun , Zou Xia , Huang Xiaoling , Liu Yin , Lu Qian , Ling Li
Published: December 2019

The quality of family relationships is important for individual and family well-being. Improving family relationships is also an important goal in methadone maintenance treatment (MMT). Little is known about factors associated with the improvement of family relationships among MMT clients. This study aimed to identify factors associated with family relationship improvement in MMT. We retrospectively analyzed existing data from 2006 to 2014 at 15 MMT clinics in Guangdong, China, including 2171 subjects with 4691 follow-ups. Generalized estimating equations were used to investigate the association between living status transitions, behavior changes and family relationship improvement, with covariates controlled for. Family relationship improvement was found in 23.1% of all follow-up intervals. Participants who began living with family, living on a regular wage, and gained employment were more likely to have improved family relationships. The quality of family relationships also improved among participants who ceased contact with drug-addicted fellows, ceased drug use, and those who were sexually active. These results suggest that improvement in living status, positive changes in drug use, and sexual activity are associated with family relationship improvement and corresponding interventions may be developed to facilitate clients’ recovery.
Mechanistic Insight of Na/K-ATPase Signaling and HO-1 into Models of Obesity and Nonalcoholic Steatohepatitis
Authors: Pratt Rebecca , Lakhani Hari Vishal , Zehra Mishghan , Desauguste Rutmann , Pillai Sneha S. , Sodhi Komal
Published: December 2019

Obesity is a multifaceted pathophysiological condition that has been associated with lipid accumulation, adipocyte dysfunction, impaired mitochondrial biogenesis and an altered metabolic profile. Redox imbalance and excessive release of inflammatory mediators have been intricately linked in obesity-associated phenotypes. Hence, understanding the mechanisms of redox signaling pathways and molecular targets exacerbating oxidative stress is crucial in improving health outcomes. The activation of Na/K-ATPase/Src signaling, and its downstream pathways, by reactive oxygen species (ROS) has been recently implicated in obesity and subsequent nonalcoholic steatohepatitis (NASH), which causes further production of ROS creating an oxidant amplification loop. Apart from that, numerous studies have also characterized antioxidant properties of heme oxygenase 1 (HO-1), which is suppressed in an obese state. The induction of HO-1 restores cellular redox processes, which contributes to inhibition of the toxic milieu. The novelty of these independent mechanisms presents a unique opportunity to unravel their potential as molecular targets for redox regulation in obesity and NASH. The attenuation of oxidative stress, by understanding the underlying molecular mechanisms and associated mediators, with a targeted treatment modality may provide for improved therapeutic options to combat clinical disorders.
Increased Adenine Nucleotide Degradation in Skeletal Muscle Atrophy
Authors: Miller Spencer G. , Hafen Paul S. , Brault Jeffrey J.
Published: December 2019

Adenine nucleotides (AdNs: ATP, ADP, AMP) are essential biological compounds that facilitate many necessary cellular processes by providing chemical energy, mediating intracellular signaling, and regulating protein metabolism and solubilization. A dramatic reduction in total AdNs is observed in atrophic skeletal muscle across numerous disease states and conditions, such as cancer, diabetes, chronic kidney disease, heart failure, COPD, sepsis, muscular dystrophy, denervation, disuse, and sarcopenia. The reduced AdNs in atrophic skeletal muscle are accompanied by increased expression/activities of AdN degrading enzymes and the accumulation of degradation products (IMP, hypoxanthine, xanthine, uric acid), suggesting that the lower AdN content is largely the result of increased nucleotide degradation. Furthermore, this characteristic decrease of AdNs suggests that increased nucleotide degradation contributes to the general pathophysiology of skeletal muscle atrophy. In view of the numerous energetic, and non-energetic, roles of AdNs in skeletal muscle, investigations into the physiological consequences of AdN degradation may provide valuable insight into the mechanisms of muscle atrophy.
Characterization and Immobilization of a Novel SGNH Family Esterase (LaSGNH1) from Lactobacillus acidophilus NCFM
Authors: Le Ly Thi Huong Luu , Yoo Wanki , Jeon Sangeun , Kim Kyeong Kyu , Kim T. Doohun
Published: December 2019

The SGNH family esterases are highly effective biocatalysts due to their strong catalytic efficiencies, great stabilities, relatively small sizes, and ease of immobilization. Here, a novel SGNH family esterase (LaSGNH1) from Lactobacillus acidophilus NCFM, which has homologues in many Lactobacillus species, was identified, characterized, and immobilized. LaSGNH1 is highly active towards acetate- or butyrate-containing compounds, such as p-nitrophenyl acetate or 1-naphthyl acetate. Enzymatic properties of LaSGNH1, including thermal stability, optimum pH, chemical stability, and urea stability, were investigated. Interestingly, LaSGNH1 displayed a wide range of substrate specificity that included glyceryl tributyrate, tert-butyl acetate, and glucose pentaacetate. Furthermore, immobilization of LaSGNH1 by crosslinked enzyme aggregates (CLEAs) showed enhanced thermal stability and efficient recycling property. In summary, this work paves the way for molecular understandings and industrial applications of a novel SGNH family esterase (LaSGNH1) from Lactobacillus acidophilus.
Comparative Effects and Mechanisms of Chitosan and Its Derivatives on Hypercholesterolemia in High-Fat Diet-Fed Rats
Authors: Chiu Chen-Yuan , Yen Tsai-En , Liu Shing-Hwa , Chiang Meng-Tsan
Published: December 2019

The present study investigated and compared the effects of different molecular weights of chitosan (high molecular weight chitosan (HC) and low molecular weight chitosan (LC)) and its derivatives (chitosan oligosaccharide (CO)) on cholesterol regulation in high-fat (HF) diet-fed rats. A diet supplementation of 5% HC, 5% LC, or 5% CO for 8 weeks showed hypocholesterolemic potential in HF diet-fed rats. Unexpectedly, a 5% CO-supplemented diet exerted hepatic damage, producing increased levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and tumor necrosis factor-alpha (TNF-α). The supplementation of HC and LC, unlike CO, significantly decreased the hepatic total cholesterol (TC) levels and increased the fecal TC levels in HF diet-fed rats. The hepatic protein expression of the peroxisome proliferator-activated receptor-α (PPARα) in the HF diet-fed rats was markedly decreased, which could be significantly reversed by both HC and LC, but not CO, supplementation. Unlike the supplementation of CO, both HC and LC supplementation could effectively reverse the HF-inhibited/induced gene expressions of the low-density lipoprotein receptor (LDLR) and cholesterol 7α-hydroxylase (CYP7A1), respectively. The upregulated intestinal acyl-CoA cholesterol acyltransferase 2 (ACAT2) protein expression in HF diet-fed rats could be reversed by HC and LC, but not CO, supplementation. Taken together, a supplementation of 5% CO in HF diet-fed rats may exert liver damage via a higher hepatic cholesterol accumulation and a higher intestinal cholesterol uptake. Both HC and LC effectively ameliorated the hypercholesterolemia and regulated cholesterol homeostasis via the activation and inhibition of hepatic (AMPKα and PPARα) and intestinal (ACAT2) cholesterol-modulators, respectively, as well as the modulation of downstream signals (LDLR and CYP7A1).
The Impact of Glycosylation of Osteopontin on Urinary Stone Formation
Authors: Anan Go , Yoneyama Tohru , Noro Daisuke , Tobisawa Yuki , Hatakeyama Shingo , Sutoh Yoneyama Mihoko , Yamamoto Hayato , Imai Atsushi , Iwamura Hiromichi , Kohada Yuki , Mikami Jotaro , Ito Jun , Kaiho Yasuhiro , Yoneyama Takahiro , Hashimoto Yasuhiro , Sato Makoto , Ohyama Chikara
Published: December 2019

Osteopontin (OPN) is a matrix glycoprotein of urinary calculi. This study aims to identify the role of aberrant glycosylation of OPN in urolithiasis. We retrospectively measured urinary glycosylated OPN normalized by urinary full-length-OPN levels in 110 urolithiasis patients and 157 healthy volunteers and 21 patients were prospectively longitudinal follow-up during stone treatment. The urinary full-length-OPN levels were measured using enzyme-linked immunosorbent assay and glycosylated OPN was measured using a lectin array and lectin blotting. The assays were evaluated using the area under the receiver operating characteristics curve to discriminate stone forming urolithiasis patients. In the retrospective cohort, urinary Gal3C-S lectin reactive- (Gal3C-S-) OPN/full-length-OPN, was significantly higher in the stone forming urolithiasis patients than in the healthy volunteers (p < 0.0001), with good discrimination (AUC, 0.953), 90% sensitivity, and 92% specificity. The Lycopersicon esculentum lectin analysis of urinary full-length-OPN showed that urinary full-length-OPN in stone forming urolithiasis patients had a polyLacNAc structure that was not observed in healthy volunteers. In the prospective longitudinal follow-up study, 92.8% of the stone-free urolithiasis group had Gal3C-S-OPN/full-length-OPN levels below the cutoff value after ureteroscopic lithotripsy (URS), whereas 71.4% of the residual-stone urolithiasis group did not show decreased levels after URS. Therefore, Gal3C-S-OPN/full-length-OPN levels could be used as a urolithiasis biomarker.
Recent Developments in Nanofiber Fabrication and Modification for Bone Tissue Engineering
Authors: Udomluck Nopphadol , Koh Won-Gun , Lim Dong-Jin , Park Hansoo
Published: December 2019

Bone tissue engineering is an alternative therapeutic intervention to repair or regenerate lost bone. This technique requires three essential components: stem cells that can differentiate into bone cells, growth factors that stimulate cell behavior for bone formation, and scaffolds that mimic the extracellular matrix. Among the various kinds of scaffolds, highly porous nanofibrous scaffolds are a potential candidate for supporting cell functions, such as adhesion, delivering growth factors, and forming new tissue. Various fabricating techniques for nanofibrous scaffolds have been investigated, including electrospinning, multi-axial electrospinning, and melt writing electrospinning. Although electrospun fiber fabrication has been possible for a decade, these fibers have gained attention in tissue regeneration owing to the possibility of further modifications of their chemical, biological, and mechanical properties. Recent reports suggest that post-modification after spinning make it possible to modify a nanofiber’s chemical and physical characteristics for regenerating specific target tissues. The objectives of this review are to describe the details of recently developed fabrication and post-modification techniques and discuss the advanced applications and impact of the integrated system of nanofiber-based scaffolds in the field of bone tissue engineering. This review highlights the importance of nanofibrous scaffolds for bone tissue engineering.
Effect of trans-Cinnamaldehyde on Methicillin-Resistant Staphylococcus aureus Biofilm Formation: Metabolic Activity Assessment and Analysis of the Biofilm-Associated Genes Expression
Authors: Kot Barbara , Sytykiewicz Hubert , Sprawka Iwona , Witeska Małgorzata
Published: December 2019

The effects of trans-cinnamaldehyde (TC) on transcriptional profiles of biofilm-associated genes and the metabolic activity of two methicillin-resistant Staphylococcus aureus (MRSA) strains showing a different degree of adherence to polystyrene, were evaluated. Metabolic activity of S. aureus in biofilm was significantly decreased in the presence of TC at 1/2 minimum biofilm inhibition concentration (MBIC). Expression levels of the genes encoding laminin binding protein (eno), elastin binding protein (ebps) and fibrinogen binding protein (fib) in the presence of TC at 1/2 MBIC were lower than in untreated biofilm in both the weakly and strongly adhering strain. The highest decrease of expression level was observed in case of fib in the strongly adhering strain, in which the amount of fib transcript was 10-fold lower compared to biofilm without TC. In the presence of TC at 1/2 MBIC after 3, 6, 8 and 12 h, the expression level of icaA and icaD, that are involved in the biosynthesis of polysaccharide intercellular adhesin, was above half lower in the weakly adhering strain compared to biofilm without TC. In the strongly adhering strain the highest decrease in expression of these genes was observed after 3 and 6 h. This study showed that TC is a promising anti-biofilm agent for use in MRSA biofilm-related infections.
Characterization and High-Level Periplasmic Expression of Thermostable α-Carbonic Anhydrase from Thermosulfurimonas Dismutans in Escherichia Coli for CO2 Capture and Utilization
Authors: Jo Byung Hoon , Hwang In Seong
Published: December 2019

Carbonic anhydrase (CA) is a diffusion-controlled enzyme that rapidly catalyzes carbon dioxide (CO2) hydration. CA has been considered as a powerful and green catalyst for bioinspired CO2 capture and utilization (CCU). For successful industrial applications, it is necessary to expand the pool of thermostable CAs to meet the stability requirement under various operational conditions. In addition, high-level expression of thermostable CA is desirable for the economical production of the enzyme. In this study, a thermostable CA (tdCA) of Thermosulfurimonas dismutans isolated from a deep-sea hydrothermal vent was expressed in Escherichia coli and characterized in terms of expression level, solubility, activity and stability. tdCA showed higher solubility, activity, and stability compared to those of CA from Thermovibrio ammonificans, one of the most thermostable CAs, under low-salt aqueous conditions. tdCA was engineered for high-level expression by the introduction of a point mutation and periplasmic expression via the Sec-dependent pathway. The combined strategy resulted in a variant showing at least an 8.3-fold higher expression level compared to that of wild-type tdCA. The E. coli cells with the periplasmic tdCA variant were also investigated as an ultra-efficient whole-cell biocatalyst. The engineered bacterium displayed an 11.9-fold higher activity compared to that of the recently reported system with a halophilic CA. Collectively these results demonstrate that the highly expressed periplasmic tdCA variant, either in an isolated form or within a whole-cell platform, is a promising biocatalyst with high activity and stability for CCU applications.
Expression and Function of Host Defense Peptides at Inflammation Sites
Authors: Prasad Suhanya V. , Fiedoruk Krzysztof , Daniluk Tamara , Piktel Ewelina , Bucki Robert
Published: December 2019

There is a growing interest in the complex role of host defense peptides (HDPs) in the pathophysiology of several immune-mediated inflammatory diseases. The physicochemical properties and selective interaction of HDPs with various receptors define their immunomodulatory effects. However, it is quite challenging to understand their function because some HDPs play opposing pro-inflammatory and anti-inflammatory roles, depending on their expression level within the site of inflammation. While it is known that HDPs maintain constitutive host protection against invading microorganisms, the inducible nature of HDPs in various cells and tissues is an important aspect of the molecular events of inflammation. This review outlines the biological functions and emerging roles of HDPs in different inflammatory conditions. We further discuss the current data on the clinical relevance of impaired HDPs expression in inflammation and selected diseases.
Differences in Stability of Viral and Viral-Cellular Fusion Transcripts in HPV-Induced Cervical Cancers
Authors: Ehrig Franziska , Häfner Norman , Driesch Corina , Kraus Christiansen Irene , Beer Katrin , Schmitz Martina , Runnebaum Ingo B. , Dürst Matthias
Published: December 2019

HPV-DNA integration results in dysregulation of viral oncogene expression. Because viral-cellular fusion transcripts inherently lack the viral AU-rich elements of the 3’UTR, they are considered to be more stable than episome-derived transcripts. The aim of this study is to provide formal proof for this assumption by comparing the stability of viral early transcripts derived from episomal and integrated HPV16 DNA, respectively. Full-length cDNA of three fusion transcripts comprising viral and cellular sequences in sense orientation were amplified and cloned into the adeno-viral-vector pAd/CMV/V5-DEST. The most abundant HPV16 oncogene transcript E6*I-E7-E1vE4-E5 with and without 3’UTR, served as reference and control, respectively. Human primary keratinocytes were transduced using high titer virus stocks. qRT-PCR was performed to determine mRNA stability in relation to GAPDH in the presence of actinomycin-D. In four independent transduction experiments, all three viral-cellular fusion transcripts were significantly more stable compared to the episome-derived reference. Among the three viral-cellular fusion transcripts the most stable transcript was devoid of the instability core motif “AUUUA”. Unexpectedly, there was no significant difference in the stability between the episome-derived transcripts either with or without 3’UTR, indicating that the AU-rich elements of the 3’UTR are not contributing to RNA stability. Instead, the three “AUUUA” motifs located in the untranslated region between the viral E4 and E5 genes may be responsible for the instability. This is the first report showing that authentic viral-cellular fusion transcripts are more stable than episome-derived transcripts. The longer half-life of the fusion transcripts may result in increased levels of viral oncoproteins and thereby drive the carcinogenic process.
Characterization of Melanoma Cell Lines Resistant to Vemurafenib and Evaluation of Their Responsiveness to EGFR- and MET-Inhibitor Treatment
Authors: Dratkiewicz Ewelina , Simiczyjew Aleksandra , Pietraszek-Gremplewicz Katarzyna , Mazurkiewicz Justyna , Nowak Dorota
Published: December 2019

Constitutively active mutated BRAF kinase occurs in more than 40% of patients suffering from melanoma. To block its activity, a specific inhibitor, vemurafenib, is applied as a therapy. Unfortunately, patients develop resistance to this drug rather quickly. Previously, we demonstrated that pairs of inhibitors directed against EGFR (epidermal growth factor receptor) and MET (hepatocyte growth factor receptor) trigger a synergistic cytotoxic effect in human melanoma cells, and decrease their invasive abilities. In this study, we aimed to generate and characterize melanoma cells resistant to vemurafenib treatment, and then to evaluate the effectiveness of a previously developed therapy in this model. We showed that melanoma cells resistant to the BRAF inhibitor are characterized by a lower proliferation rate and they acquire a spindle-like shape. Using Western Blot, we also noticed increased levels of EGFR, MET, and selected markers of cancer stem cells in generated cell lines. Resistant cells also exhibited increased invasive abilities and elevated proteolytic activity, observed using scratch wound assays and gelatin zymography. Moreover, combination therapy reduced their viability, as measured with a colorimetric cytotoxicity test, and decreased invasiveness. The obtained results validate the application of combination therapy directed against EGFR and MET in melanoma cells resistant to treatment with inhibitors of mutated BRAF.
Neurochemical Characterization of Neurons Expressing Estrogen Receptor β in the Hypothalamic Nuclei of Rats Using in Situ Hybridization and Immunofluorescence
Authors: Kanaya Moeko , Higo Shimpei , Ozawa Hitoshi
Published: December 2019

Estrogens play an essential role in multiple physiological functions in the brain, including reproductive neuroendocrine, learning and memory, and anxiety-related behaviors. To determine these estrogen functions, many studies have tried to characterize neurons expressing estrogen receptors known as ERα and ERβ. However, the characteristics of ERβ-expressing neurons in the rat brain still remain poorly understood compared to that of ERα-expressing neurons. The main aim of this study is to determine the neurochemical characteristics of ERβ-expressing neurons in the rat hypothalamus using RNAscope in situ hybridization (ISH) combined with immunofluorescence. Strong Esr2 signals were observed especially in the anteroventral periventricular nucleus (AVPV), bed nucleus of stria terminalis, hypothalamic paraventricular nucleus (PVN), supraoptic nucleus, and medial amygdala, as previously reported. RNAscope ISH with immunofluorescence revealed that more than half of kisspeptin neurons in female AVPV expressed Esr2, whereas few kisspeptin neurons were found to co-express Esr2 in the arcuate nucleus. In the PVN, we observed a high ratio of Esr2 co-expression in arginine-vasopressin neurons and a low ratio in oxytocin and corticotropin-releasing factor neurons. The detailed neurochemical characteristics of ERβ-expressing neurons identified in the current study can be very essential to understand the estrogen signaling via ERβ.
Radioprotective and Antimutagenic Effects of Pycnanthus angolensis Warb Seed Extract against Damage Induced by X rays
Authors: Achel Daniel Gyingiri , Alcaraz-Saura Miguel , Castillo Julián , Olivares Amparo , Alcaraz Miguel
Published: December 2019

Although different studies have demonstrated different applications of Pycnanthus angolensis extracts in traditional African and Asian medicine, its possible antimutagenic or genoprotective capacities have never been explored. We studied these capabilities of Pycnanthus angolensis seed extract (PASE) by means of the two micronucleus assays, determining the frequency of micronucleus (MN) yield in mouse bone marrow (in vivo) and in human lymphocytes blocked by cytochalasin B (in vitro). PASE exhibited a significant genoprotective capacity (p < 0.001) against X-rays with a protection factor of 35% in both in vivo and in vitro assays. Further, its radioprotective effects were determined by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-tetrazolium bromide (MTT) cell viability test in two cell lines: one being radiosensitive (i.e., human prostate epithelium (PNT2) cells) and the other being radioresistant (i.e., B16F10 melanoma cells). In the radiosensitive cells, PASE showed a protection factor of 35.5%, thus eliminating 43.8% of X-ray-induced cell death (p < 0.001) and a dose reduction factor of 2.5. In the radioresistant cells, a protection factor of 29% (p < 0.001) with a dose reduction factor of 4 was realized. PASE elicited a greater radioprotective capacity than the substances currently used in radiation oncology and, thus, could be developed as a nutraceutical radioprotectant for workers and patients exposed to ionizing radiation.
Correlation of Micro-Computed Tomography Assessment of Valvular Mineralisation with Histopathological and Immunohistochemical Features of Calcific Aortic Valve Disease
Authors: Solache-Berrocal Guillermo , Barral-Varela Ana María , Areces-Rodríguez Sheila , Junco-Vicente Alejandro , Vallina-Álvarez Aitana , Corte-Torres María Daniela , Valdivielso José Manuel , Llosa Juan Carlos , Morís César , Martín María , Rodríguez Isabel
Published: December 2019

Aortic valve stenosis is a serious disease with increasing prevalence in developed countries. Research aimed at uncovering the molecular mechanisms behind its main cause, aortic valve calcification, is thus crucial for the development of future therapies. It is frequently difficult to measure the extent of mineralisation in soft tissues and some methods require the destruction of the sample. Micro-computed tomography (µCT), a non-destructive technique, was used to quantify the density and volume of calcium deposits on cusps from 57 explanted aortic valves. Conventional and immunostaining techniques were used to characterise valve tissue degeneration and the inflammatory and osteogenic stage with several markers. Although most of the analysed cusps came from severe stenosis patients, the µCT parameter bone volume/tissue volume ratio distinguished several degrees of mineralisation that correlated with the degree of structural change in the tissue and the amount of macrophage infiltration as determined by CD68 immunohistochemistry. Interestingly, exosomal markers CD63 and Alix co-localised with macrophage infiltration surrounding calcium deposits, suggesting that those vesicles could be produced at least in part by these immune cells. In conclusion, we have shown that the ex vivo assessment of aortic valve mineralisation with µCT reflects the molecular and cellular changes in pathological valves during progression towards stenosis. Thus, our results give additional validity to quantitative μCT as a convenient laboratory tool for basic research on this type of cardiovascular calcification.