bluuebunny/crossref_metadata_2025 huggingface dataset:
Psychometric Testing of the Korean Version of the Attitudes toward the Advance Directives in Low-Income Chronically Ill Older Adults
Authors: Kim JinShil , Heo Seongkum , Hong Sun Woo , Kim HeeRyang , Jung Ahrang , An Minjeong , Shim JaeLan
Published: March 2020

The purpose of this study was to examine the psychometric properties of the Advance Directive Attitude Survey in Korean (K-ADAS), a measure of attitudes toward advance directives (ADs). A total of 118 low-income, community-dwelling older adults (mean age, 75.09 years) participated. An exploratory factor analysis (EFA) was conducted to determine the factor structure of the K-ADAS. Validity was further assessed by known associations of the K-ADAS with perceived susceptibility and severity using part of the Advance Care Planning surveys. Its reliability was examined by calculating alpha coefficients. EFA determined a three-factor structure model with good model fit. Validity was further supported with significant correlations between the K-ADAS and susceptibility and severity. Reliability was supported by adequate level of Cronbach’s alpha. The K-ADAS was a valid and reliable measure for assessment of AD attitudes with a sound model fit. Thus, the K-ADAS can be used to assess AD attitudes among community-dwelling elders.
Development of General Exposure Factors for Risk Assessment in Korean Children
Authors: Yoon Hyojung , Yoo Sun-Kyoung , Seo Jungkwan , Kim Taksoo , Kim Pyeongsoon , Kim Pil-Je , Park Jinhyeon , Heo Jung , Yang Wonho
Published: March 2020

There has been an increasing need for the risk assessment of external environmental hazards in children because they are more sensitive to hazardous chemical exposure than adults. Therefore, the development of general exposure factors is required for appropriate risk assessment in Korean children. This study aimed to determine the general exposure factors among Korean children aged ≤18 years. We developed the recommended exposure factors across five categories: physiological variables, inhalation rates, food and drinking water intake, time–activity patterns, and soil and dust ingestion. National databases were used, and direct measurements and questionnaire surveys of representative samples were performed to calculate the inhalation rate, water intake, and soil ingestion rate. With regard to the time–activity patterns, the daily inhalation rates ranged from 9.49 m3/day for children aged 0–2 years to 14.98 m3/day for those aged 16–18 years. This study found that Korean children spent an average of 22.64 h indoors, 0.63 h outdoors, and 0.73 h in-transit on weekdays. The general exposure factors of Korean children were studied for the first time, and these results could be used to assess children’s exposure and risk. They also suggest the differences compared with the results of international results.
Analysis of Heavy Metal Contamination of Agricultural Soils and Related Effect on Population Health—A Case Study for East River Basin in China
Authors: He Liping , Hu Wei , Wang Xiaofeng , Liu Yu , Jiang Yan , Meng Yanbin , Xiao Qipeng , Guo Xinxin , Zhou Yanfeng , Bi Yongyi , Lu Yuanan
Published: March 2020

To understand the heavy metal pollution in agricultural soils along the East River basin and assess the pollution related health effect to local residents, interviews and archived data were obtained to identify the study sites affected by polluted tailing. Soil samples were collected and tested for heavy metal content and the Comprehensive Pollution Index (CPI). The degree of pollution of agricultural soils in the area was assessed using GIS-based Spatial distribution map of heavy metals and the trend of soil heavy metal risk. Two villages (Matian and Zhudui) near the East River were included in this study for health effect assessment. A total of 193 residents aged 15 or above from each village were tested for the present status of chronic diseases. Convenient sampling method was used to collect blood samples from 78 residents for heavy metal concentration. The contents of Pb, Cd, As, Zn, and Cu in the agricultural soils were all over the standards with a moderate to severe CPI. Among these metals, Cd was the highest followed by Pb, and Cu was the lowest. The contents of Pb, Cd, As, and Zn tend to be higher in soils closer to the river. The prevalence of chronic diseases was over 30%, which is significantly higher than the report from the national central region (23.15%). The average blood lead level (BLL) among children under 14 years is 7.42 μg/dL. Although the adults in Matian village had a significantly higher BLL (χ2 = 8.70, p = 0.03) as compared to Zhudui village, there was no significant difference for the prevalence of chronic diseases between the two villages (χ2 = 3.23, p = 0.09). The mean BLL of children and the proportion of children with BLL ≥ 10 µg/dL in this study are equivalent to the national average. The higher BLL concentration and prevalence of chronic diseases in adults might be due to their long-term exposure to heavy metal contamination environment and higher background level of heavy metals. Findings from this study will form the baseline information for local government to the development of effective approaches to control the heavy metal contamination and reduce the pollution related adverse health effect on local residents.
The Impact of Social Support on the Health of the Rural Elderly in China
Authors: Bai Yunli , Bian Fang , Zhang Linxiu , Cao Yueming
Published: March 2020

With the dramatic trend of global aging, the physical and mental health of the rural elderly has attracted significant attention. Social support plays an important role in improving the health of the elderly. However, assessing the impact of social support on the physical and mental health of the elderly is challenging in rural China. This paper analyzes the impact of social support on the physical and mental health of the Chinese rural elderly based on data collected from households and village cadres. Probit, Oprobit, and ordinary least squares (OLS) are used to estimate these effects. The results show that 24.3% of the rural elderly are in bad physical health, and 32.9% of them are depressed. Physical and mental health is worse among the female elderly and among those who are in western provinces. Having pensions, taking care of grandchildren, and communicating with children by telephone are shown to significantly improve the mental health of the rural elderly. The government could promote the mental health of the rural elderly by improving public health services, increasing pensions, providing free mobile phones to elderly people in poverty, and advocating that the younger generation provide emotional support.
Neonicotinoids: Spreading, Translocation and Aquatic Toxicity
Authors: Mörtl Mária , Vehovszky Ágnes , Klátyik Szandra , Takács Eszter , Győri János , Székács András
Published: March 2020

Various environmental and ecotoxicological aspects related to applications of neonicotinoid insecticides are assessed. Dosages of neonicotinoids applied in seed coating materials were determined and are compared to other applications (spray and granule). Environmental levels in soils and affecting factors in translocation are discussed. Excretion of neonicotinoids via guttation from coated maize seeds up to two months upon emergence, as well as cross-contamination of plants emerged from non-coated seeds or weeds nearby have been demonstrated. Contamination of surface waters is discussed in scope of a worldwide review and the environmental fate of the neonicotinoid active ingredients and the formulating surfactant appeared to be mutually affected by each other. Toxicity of neonicotinoid active ingredients and formulations on Daphnia magna completed with some investigations of activity of the detoxifying glutathione S-transferase enzyme demonstrated the modified toxicity due to the formulating agents. Electrophysiological results on identified central neurons of the terrestrial snail Helix pomatia showed acetylcholine antagonist (inhibitory) effects of neonicotinoid insecticide products, but no agonist (ACh-like) effects were recorded. These data also suggested different molecular targets (nicotinergic acetylcholine receptors and acetylcholine esterase enzyme) of neonicotinoids in the snail central nervous system.
A Cost-Minimization Analysis of a Medical Record-based, Store and Forward and Provider-to-provider Telemedicine Compared to Usual Care in Catalonia: More Agile and Efficient, Especially for Users
Authors: López Seguí Francesc , Franch Parella Jordi , Gironès García Xavier , Mendioroz Peña Jacobo , García Cuyàs Francesc , Adroher Mas Cristina , García-Altés Anna , Vidal-Alaball Josep
Published: March 2020

Background: Telemedicine (interconsultation between primary and hospital care teams) has been operating in the counties of Central Catalonia Bages, Moianès and Berguedà since 2011, specializing in teledermatology, teleulcers, teleophthalmology and teleaudiometries. For the period until the end of 2019, a total of 52,198 visits were recorded. Objective: To analyze the differential costs between telemedicine and usual care in a semi-urban environment. Methodology: A cost-minimization evaluation, including direct and indirect costs from a societal perspective, distinguishing healthcare and user’s costs, was carried out over a three-month period. Results: Telemedicine saved € 780,397 over the period analyzed. A differential cost favorable to telemedicine of about € 15 per visit was observed, with the patient being the largest beneficiary of this saving (by 85%) in terms of shorter waiting times and travel costs. From the healthcare system perspective, moving the time spent in a hospital care consultation to primary care is efficient in terms of the total time devoted per patient. In social terms and in this context, telemedicine is more efficient than usual care. Conclusion: Allowing users to save time in terms of consultation and travel is the main driver of interconsultation between primary and hospital care savings in a semi-urban context. The telemedicine service is also economically favorable for the healthcare system, enabling it to provide a more agile service, which also benefits healthcare professionals.
Intracranial Vertebrobasilar Calcification in Patients with Ischemic Stroke Is a Predictor of Recurrent Stroke, Vascular Disease, and Death: A Case-Control Study
Authors: Magdič Jožef , Cmor Nino , Kaube Matevž , Hojs Fabjan Tanja , Hauer Larissa , Sellner Johann , Pikija Slaven
Published: March 2020

Intracranial artery calcification can be detected on nonenhanced brain computer tomography (NECT) and is a predictor of early vascular events. Here, we assessed the impact of vertebrobasilar artery calcification (VBC) on the long-term risk for recurrent stroke and vascular events. We performed a case-control trial of all consecutive stroke patients admitted to the University Hospital of Maribor, Slovenia over a period of 14 months. VBC was defined as presence of a hyperdense area within vertebrobasilar arteries that exceeds > 90 Hounsfield units as seen on NECT. Clinical follow-up information was obtained from the hospital documentation system and mortality registry of the district and included recurrent stroke, subsequent vascular events (myocardial infarction, heart failure, peripheral arterial occlusive disease), and death. We followed a total of 448 patients for a median of 1505 days (interquartile range, IQR 188-2479). Evidence for VBC was present in 243 (54.2%) patients. Median age was 76 years, recurrent stroke occurred in 33 (7.4%), any vascular events in 71 (15.8%), and death in 276 (61.6%). VBC was associated with a higher risk of recurrent stroke (hazard ratio, HR 3.13, 95% confidence interval (CI 1.35–7.20)) and vascular events (HR 2.05, 95% CI 1.21–3.47). Advanced age, male gender, and ischemic stroke involving the entire anterior circulation raised the likelihood for death. We conclude that the presence of VBC in patients with ischemic stroke is a short- and long-term prognostic factor for stroke recurrence and subsequent manifestation of acute vascular disease. Further understanding of the pathophysiology of VBC is warranted.
A Scoping Review of Children and Adolescents’ Active Travel in Ireland
Authors: Costa João , Adamakis Manolis , O’Brien Wesley , Martins João
Published: March 2020

There appears to be a lack of existing data that comprehensively summarizes the evidence of children and adolescents’ active travel in the Republic of Ireland. In lieu of this, a scoping review was conducted to map the existing literature (2000–2020) on children and adolescents’ active travel in the Republic of Ireland. A scoping review design extracted a total of 19 publications, which show a consistent focus on the identified population’s active travel patterns, mainly to and from school, mostly self-report and cross-sectional research study designs; however, there are few longitudinal data, intervention and participatory studies. Key issues from these identified scoping review studies are discussed with the potential to better inform policy makers, practitioners and researchers to delineate programmes and strategies for promoting active travel among children and adolescents in the Republic of Ireland.
Reconnaissance of Surface Water Estrogenicity and the Prevalence of Intersex in Smallmouth Bass (Micropterus Dolomieu) Inhabiting New Jersey
Authors: Iwanowicz Luke R. , Smalling Kelly L. , Blazer Vicki S. , Braham Ryan P. , Sanders Lakyn R. , Boetsma Anna , Procopio Nicholas A. , Goodrow Sandra , Buchanan Gary A. , Millemann Daniel R. , Ruppel Bruce , Vile John , Henning Brian , Abatemarco John
Published: March 2020

The observation of testicular oocytes in male fishes has been utilized as a biomarker of estrogenic endocrine disruption. A reconnaissance project led in the Northeastern United States (US) during the period of 2008–2010 identified a high prevalence of intersex smallmouth bass on or near US Fish & Wildlife Service National Wildlife Refuges that included the observation of 100% prevalence in smallmouth bass males collected from the Wallkill River, NJ, USA. To better assess the prevalence of intersex smallmouth bass across the state of New Jersey, a tiered reconnaissance approach was initiated during the fall of 2016. Surface water samples were collected from 101 (85 river, 16 lake/reservoir) sites across the state at base-flow conditions for estrogenicity bioassay screening. Detectable estrogenicity was observed at 90% of the sites and 64% were above the US Environmental Protection Agency trigger level of 1 ng/L. Median surface water estrogenicity was 1.8 ng/L and a maximum of 6.9 ng/L E2EqBLYES was observed. Adult smallmouth bass were collected from nine sites, pre-spawn during the spring of 2017. Intersex was identified in fish at all sites, and the composite intersex prevalence was 93.8%. Prevalence across sites ranged from 70.6% to 100%. In addition to intersex, there was detectable plasma vitellogenin in males at all sites. Total estrogenicity in surface water was determined at these fish collection sites, and notable change over time was observed. Correlation analysis indicated significant positive correlations between land use (altered land; urban + agriculture) and surface water estrogenicity. There were no clear associations between land use and organismal metrics of estrogenic endocrine disruption (intersex or vitellogenin). This work establishes a baseline prevalence of intersex in male smallmouth bass in the state of New Jersey at a limited number of locations and identifies a number of waterbodies with estrogenic activity above an effects-based threshold.
Understanding Alternatives to Tobacco Production in Kenya: A Qualitative Analysis at the Sub-National Level
Authors: Clark Madelyn , Magati Peter , Drope Jeffrey , Labonte Ronald , Lencucha Raphael
Published: March 2020

Tobacco is a key cash crop for many farmers in Kenya, although there is a variety of challenges associated with tobacco production. This study seeks to understand alternatives to tobacco production from the perspective of government officials, extension officers, and farmers at the sub-national level (Migori, Busia, and Meru) in Kenya. The study analyzes data from qualitative key-informant interviews with government officials and extension officers (n = 9) and focus group discussions (FGDs) with farmers (n = 5). Data were coded according to pre-identified categories derived from the research aim, namely, opportunities and challenges of tobacco farming and alternative crops, as well findings that illustrate the policy environment that shapes the agricultural context in these regions. We highlight important factors associated with the production of non-tobacco agricultural commodities, including the factors that shape the ability of these non-agricultural commodities to serve as viable alternatives to tobacco. The results highlight the effect that several factors, including access to capital, markets, and governmental assistance, have on farmer decisions. The results additionally display the structured policy approaches that are being promoted in governmental offices towards agricultural production, as well as the institutional shortcomings that inhibit their implementation at the sub-national level.
Malocclusion, Dental Caries and Oral Health-Related Quality of Life: A Comparison between Adolescent School Children in Urban and Rural Regions in Peru
Authors: Cadenas de Llano-Pérula Maria , Ricse Estela , Fieuws Steffen , Willems Guy , Orellana-Valvekens Maria Fernanda
Published: March 2020

Rural, isolated areas benefit less from caries prevention programs and access to treatment than urban areas, and, hence, differences in oral health can be expected. This study aims to assess the prevalence of caries and malocclusion in urban and rural areas of Peru and to compare them with patients’ oral health perception. A total of 1062 adolescents were examined in five schools of rural (Titicaca) and urban (Lima and Cuzco) areas in Peru. Decay Missing Filled Teeth’s Surfaces, the Index of Complexity, Outcome and Need and the Child Oral Health Impact Profile short form-19 (COHIP-SF 19) were used to assess caries, severity of malocclusion and Oral Health Quality of Life, respectively. Significant differences in the prevalence (p = 0.001) and degree of caries (p = 0.001) were found between regions. The prevalence of caries was the highest in Cuzco (97.65%), followed by Titicaca (88.81%) and Lima (76.42%). The severity of malocclusion was the lowest in Titicaca. There was a negative relation between malocclusion, caries and COHIP-SF 19, with no evidence of a difference between the regions. This suggests that the higher the prevalence of caries and the more severe the malocclusion, the poorer the perception of oral health. In our study, rural areas presented a lower severity of malocclusion than urban areas.
How Is the Self-Perceived Work Ability Affected by the Duration of Unemployment, Heart Rate Variability and the Amount of Physical Activity in Older Long-Term Unemployed Persons?
Authors: Bumann Anke , Niederer Daniel , Santarossa Constanze , Banzer Winfried , Vogt Lutz
Published: March 2020

Background: This study investigated whether work ability is associated with the duration of unemployment, heart rate variability (HRV), and the level of physical activity. Methods: Thirty-four unemployed persons (mean 55.7 ± standard deviation 33.3 years, 22 female, 12 male, unemployed: range 1–22.5 years) participated in the cross-sectional study. The Work Ability Index (WAI) and International Physical Activity Questionnaire (IPAQ) were applied. Short-term (five minutes) resting HRV (Low Frequency (LF), High Frequency (HF), Total Power (TP)) was collected. Results: Work ability was positively associated with the HRV: LF (r = 0.383; p = 0.025), HF (r = 0.412; p = 0.015) and TP (r = 0.361; p = 0.036). The WAI showed a positive linear correlation with the amount of total physical activity (r = 0.461; p = 0.006) as well as with the amount of moderate to vigorous physical activity (r = 0.413; p = 0.015). No association between the WAI and the duration of unemployment occurred. Conclusions: the relation between self-perceived work ability, health-associated parameters, the HRV and the level of physical activity points out the relevance of health-care exercise and the need of stress-reducing interventions to improve perceived work ability. Our results point out the need for the further and more holistic development of healthcare for the unemployed.
Ocular Cell Lines and Genotoxicity Assessment
Authors: Souto Eliana B. , Campos Joana R. , Da Ana Raquel , Martins-Gomes Carlos , Silva Amélia M. , Souto Selma B. , Lucarini Massimo , Durazzo Alessandra , Santini Antonello
Published: March 2020

Genotoxicity screening tests aim to evaluate if and to what extent a compound in contact with the human body (e.g., a drug molecule, a compound from the environment) interacts with DNA. The comet assay is a sensitive method used to predict the risk of DNA damage in individual cells, as it quantifies the tape breaks, being the alkaline version (pH > 13) the most commonly used in the laboratory. Epithelial cells serve as biomatrices in genotoxicity assessments. As ca. 80% of solid cancers are of epithelial origin, the quantification of the DNA damage upon exposure of epithelial cells to a drug or drug formulation becomes relevant. Comet assays run in epithelial cells also have clinical applications in human biomonitoring, which assesses whether and to what extent is the human body exposed to environmental genotoxic compounds and how such exposure changes over time. Ocular mucosa is particularly exposed to environmental assaults. This review summarizes the published data on the genotoxicity assessment in estimating DNA damage in epithelial cells with a special focus on ocular cell lines. General comet assay procedures for ex vivo and in vivo epithelium samples are also described.
Pharmacological Mechanisms Underlying the Neuroprotective Effects of Alpinia oxyphylla Miq. on Alzheimer’s Disease
Authors: Xu Jia , Wang Fang , Guo Jiejie , Xu Chunshuang , Cao Yanzi , Fang Zhiling , Wang Qinwen
Published: March 2020

Alpinia oxyphylla Miq. (i.e., A. oxyphylla), a traditional Chinese medicine, can exert neuroprotective effects in ameliorating mild cognitive impairment and improving the pathological hallmarks of Alzheimer’s disease (AD). Here, 50 active compounds and 164 putative targets were collected and identified with 251 clinically tested AD-associated target proteins using network pharmacology approaches. Based on the Gene Ontology/Kyoto Encyclopedia of Genes and Genomes pathway enrichments, the compound-target-pathway-disease/protein–protein interaction network constructions, and the network topological analysis, we concluded that A. oxyphylla may have neuroprotective effects by regulating neurotransmitter function, as well as brain plasticity in neuronal networks. Moreover, closely-related AD proteins, including the amyloid-beta precursor protein, the estrogen receptor 1, acetylcholinesterase, and nitric oxide synthase 2, were selected as the bottleneck nodes of network for further verification by molecular docking. Our analytical results demonstrated that terpene, as the main compound of A. oxyphylla extract, exerts neuroprotective effects, providing new insights into the development of a natural therapy for the prevention and treatment of AD.
Loss of ISWI ATPase SMARCA5 (SNF2H) in Acute Myeloid Leukemia Cells Inhibits Proliferation and Chromatid Cohesion
Authors: Zikmund Tomas , Paszekova Helena , Kokavec Juraj , Kerbs Paul , Thakur Shefali , Turkova Tereza , Tauchmanova Petra , Greif Philipp A. , Stopka Tomas
Published: March 2020

ISWI chromatin remodeling ATPase SMARCA5 (SNF2H) is a well-known factor for its role in regulation of DNA access via nucleosome sliding and assembly. SMARCA5 transcriptionally inhibits the myeloid master regulator PU.1. Upregulation of SMARCA5 was previously observed in CD34+ hematopoietic progenitors of acute myeloid leukemia (AML) patients. Since high levels of SMARCA5 are necessary for intensive cell proliferation and cell cycle progression of developing hematopoietic stem and progenitor cells in mice, we reasoned that removal of SMARCA5 enzymatic activity could affect the cycling or undifferentiated state of leukemic progenitor-like clones. Indeed, we observed that CRISPR/cas9-mediated SMARCA5 knockout in AML cell lines (S5KO) inhibited the cell cycle progression. We also observed that the SMARCA5 deletion induced karyorrhexis and nuclear budding as well as increased the ploidy, indicating its role in mitotic division of AML cells. The cytogenetic analysis of S5KO cells revealed the premature chromatid separation. We conclude that deleting SMARCA5 in AML blocks leukemic proliferation and chromatid cohesion.
Claudin-12 Knockout Mice Demonstrate Reduced Proximal Tubule Calcium Permeability
Authors: Plain Allein , Pan Wanling , O’Neill Deborah , Ure Megan , Beggs Megan R. , Farhan Maikel , Dimke Henrik , Cordat Emmanuelle , Alexander R. Todd
Published: March 2020

The renal proximal tubule (PT) is responsible for the reabsorption of approximately 65% of filtered calcium, primarily via a paracellular pathway. However, which protein(s) contribute this paracellular calcium pore is not known. The claudin family of tight junction proteins confers permeability properties to an epithelium. Claudin-12 is expressed in the kidney and when overexpressed in cell culture contributes paracellular calcium permeability (PCa). We therefore examined claudin-12 renal localization and its contribution to tubular paracellular calcium permeability. Claudin-12 null mice (KO) were generated by replacing the single coding exon with β-galactosidase from Escherichia coli. X-gal staining revealed that claudin-12 promoter activity colocalized with aquaporin-1, consistent with the expression in the PT. PTs were microperfused ex vivo and PCa was measured. PCa in PTs from KO mice was significantly reduced compared with WT mice. However, urinary calcium excretion was not different between genotypes, including those on different calcium containing diets. To assess downstream compensation, we examined renal mRNA expression. Claudin-14 expression, a blocker of PCa in the thick ascending limb (TAL), was reduced in the kidney of KO animals. Thus, claudin-12 is expressed in the PT, where it confers paracellular calcium permeability. In the absence of claudin-12, reduced claudin-14 expression in the TAL may compensate for reduced PT calcium reabsorption.
Endocrine-Disrupting Chemicals and Their Effects during Female Puberty: A Review of Current Evidence
Authors: Lucaccioni Laura , Trevisani Viola , Marrozzini Lucia , Bertoncelli Natascia , Predieri Barbara , Lugli Licia , Berardi Alberto , Iughetti Lorenzo
Published: March 2020

Puberty is the process of physical changes between childhood and adulthood during which adolescents reach sexual maturity and become capable of reproduction. It is considered one of the main temporal windows of susceptibility for the influence of the endocrine-disrupting chemicals (EDCs). EDCs may act as single chemical agents or as chemical mixtures; they can be pubertal influencers, accelerating and anticipating the processing of maturation of secondary sexual characteristics. Moreover, recent studies have started to point out how exposure to EDCs during puberty may predispose to breast cancer later in life. In fact, the estrogen-mimicking endocrine disruptors (EEDs) may influence breast tissue development during puberty in two main ways: the first is the action on the proliferation of the breast stromal cells, the second concerns epigenetic mechanisms. The aim of this mini-review was to better highlight what is new and what is not completely known regarding the role of EDCs during puberty.
Effects of Thryptophan Hydroxylase Blockade by P-Chlorophenylalanine on Contextual Memory Reconsolidation after Training of Different Intensity
Authors: Deryabina Irina B. , Andrianov Viatcheslav V. , Muranova Lyudmila N. , Bogodvid Tatiana K. , Gainutdinov Khalil L.
Published: March 2020

The processes of memory formation and its storage are extremely dynamic. Therefore, the determination of the nature and temporal evolution of the changes that underlie the molecular mechanisms of retrieval and cause reconsolidation of memory is the key to understanding memory formation. Retrieval induces the plasticity, which may result in reconsolidation of the original memory and needs critical molecular events to stabilize the memory or its extinction. 4-Chloro-DL-phenylalanine (P-chlorophenylalanine-PCPA) depresses the most limiting enzyme of serotonin synthesis the tryptophan hydroxylase. It is known that PCPA reduces the serotonin content in the brain up to 10 times in rats (see Methods). We hypothesized that the PCPA could behave the similar way in snails and could reduce the content of serotonin in snails. Therefore, we investigated the effect of PCPA injection on contextual memory reconsolidation using a protein synthesis blocker in snails after training according to two protocols of different intensities. The results obtained in training according to the first protocol using five electrical stimuli per day for 5 days showed that reminding the training environment against the background of injection of PCPA led to a significant decrease in contextual memory. At the same time, the results obtained in training according to the second protocol using three electrical stimuli per day for 5 days showed that reminding the training environment against the injection of PCPA did not result in a significant change in contextual memory. The obtain results allowed us to conclude that the mechanisms of processes developed during the reconsolidation of contextual memory after a reminding depend both on the intensity of learning and on the state of the serotonergic system.
The Rice Basic Helix–Loop–Helix 79 (OsbHLH079) Determines Leaf Angle and Grain Shape
Authors: Seo Hyoseob , Kim Suk-Hwan , Lee Byoung-Doo , Lim Jung-Hyun , Lee Sang-Ji , An Gynheung , Paek Nam-Chon
Published: March 2020

Changes in plant architecture, such as leaf size, leaf shape, leaf angle, plant height, and floral organs, have been major factors in improving the yield of cereal crops. Moreover, changes in grain size and weight can also increase yield. Therefore, screens for additional factors affecting plant architecture and grain morphology may enable additional improvements in yield. Among the basic Helix-Loop-Helix (bHLH) transcription factors in rice (Oryza sativa), we found an enhancer-trap T-DNA insertion mutant of OsbHLH079 (termed osbhlh079-D). The osbhlh079-D mutant showed a wide leaf angle phenotype and produced long grains, similar to the phenotypes of mutants with increased brassinosteroid (BR) levels or enhanced BR signaling. Reverse transcription-quantitative PCR analysis showed that BR signaling-associated genes are largely upregulated in osbhlh079-D, but BR biosynthesis-associated genes are not upregulated, compared with its parental japonica cultivar ‘Dongjin’. Consistent with this, osbhlh079-D was hypersensitive to BR treatment. Scanning electron microscopy revealed that the expansion of cell size in the adaxial side of the lamina joint was responsible for the increase in leaf angle in osbhlh079-D. The expression of cell-elongation-associated genes encoding expansins and xyloglucan endotransglycosylases/hydrolases increased in the lamina joints of leaves in osbhlh079-D. The regulatory function of OsbHLH079 was further confirmed by analyzing 35S::OsbHLH079 overexpression and 35S::RNAi-OsbHLH079 gene silencing lines. The 35S::OsbHLH079 plants showed similar phenotypes to osbhlh079-D, and the 35S::RNAi-OsbHLH079 plants displayed opposite phenotypes to osbhlh079-D. Taking these observations together, we propose that OsbHLH079 functions as a positive regulator of BR signaling in rice.
Hepatitis C Virus Entry: An Intriguingly Complex and Highly Regulated Process
Authors: Colpitts Che , Tsai Pei-Ling , Zeisel Mirjam
Published: March 2020

Hepatitis C virus (HCV) is a major cause of chronic hepatitis and liver disease worldwide. Its tissue and species tropism are largely defined by the viral entry process that is required for subsequent productive viral infection and establishment of chronic infection. This review provides an overview of the viral and host factors involved in HCV entry into hepatocytes, summarizes our understanding of the molecular mechanisms governing this process and highlights the therapeutic potential of host-targeting entry inhibitors.
Does Melatonin Exert Its Effect on Ram Sperm Capacitation Through Nitric Oxide Synthase Regulation?
Authors: Miguel-Jiménez Sara , Carvajal-Serna Melissa , Calvo Silvia , Casao Adriana , Cebrián-Pérez José Álvaro , Muiño-Blanco Teresa , Pérez-Pe Rosaura
Published: March 2020

Nitric oxide (NO·), synthesized from L-arginine by nitric oxide synthase (NOS), is involved in sperm functionality. NOS isoforms have been detected in spermatozoa from different species, and an increment in NOS activity during capacitation has been reported. This work aims to determine the presence and localization of NOS isoforms in ram spermatozoa and analyse their possible changes during in vitro capacitation. Likewise, we investigated the effect of melatonin on the expression and localization of NOS and NO· levels in capacitated ram spermatozoa. Western blot analysis revealed protein bands associated with neuronal NOS (nNOS) and epithelial NOS (eNOS) but not with inducible NOS (iNOS). However, the three isoforms were detected by indirect immunofluorescence (IFI), and their immunotypes varied over in vitro capacitation with cAMP-elevating agents. NO· levels (evaluated by DAF-2-DA/PI staining) increased after in vitro capacitation, and the presence of L-arginine in the capacitating medium raised NO· production and enhanced the acrosome reaction. Incubation in capacitating conditions with a high-cAMP medium with melatonin modified the NOS distribution evaluated by IFI, but no differences in Western blotting were observed. Melatonin did not alter NO· levels in capacitating conditions, so we could infer that its role in ram sperm capacitation would not be mediated through NO· metabolism.
ECM Characterization Reveals a Massive Activation of Acute Phase Response during FSGS
Authors: Bukosza Eva Nora , Kornauth Christoph , Hummel Karin , Schachner Helga , Huttary Nicole , Krieger Sigurd , Nöbauer Katharina , Oszwald André , Razzazi Fazeli Ebrahim , Kratochwill Klaus , Aufricht Christoph , Szénási Gabor , Hamar Peter , Gebeshuber Christoph A.
Published: March 2020

The glomerular basement membrane (GBM) and extra-cellular matrix (ECM) are essential to maintain a functional interaction between the glomerular podocytes and the fenestrated endothelial cells in the formation of the slit diaphragm for the filtration of blood. Dysregulation of ECM homeostasis can cause Focal segmental glomerulosclerosis (FSGS). Despite this central role, alterations in ECM composition during FSGS have not been analyzed in detail yet. Here, we characterized the ECM proteome changes in miR-193a-overexpressing mice, which suffer from FSGS due to suppression of Wilms’ tumor 1 (WT1). By mass spectrometry we identified a massive activation of the acute phase response, especially the complement and fibrinogen pathways. Several protease inhibitors (ITIH1, SERPINA1, SERPINA3) were also strongly increased. Complementary analysis of RNA expression data from both miR-193a mice and human FSGS patients identified additional candidate genes also mainly involved in the acute phase response. In total, we identified more than 60 dysregulated, ECM-associated genes with potential relevance for FSGS progression. Our comprehensive analysis of a murine FSGS model and translational comparison with human data offers novel targets for FSGS therapy.
In Search for Genes Related to Atherosclerosis and Dyslipidemia Using Animal Models
Authors: Poznyak Anastasia V. , Grechko Andrey V. , Wetzker Reinhard , Orekhov Alexander N.
Published: March 2020

Atherosclerosis is a multifactorial chronic disease that affects large arteries and may lead to fatal consequences. According to current understanding, inflammation and lipid accumulation are the two key mechanisms of atherosclerosis development. Animal models based on genetically modified mice have been developed to investigate these aspects. One such model is low-density lipoprotein (LDL) receptor knockout (KO) mice (ldlr−/−), which are characterized by a moderate increase of plasma LDL cholesterol levels. Another widely used genetically modified mouse strain is apolipoprotein-E KO mice (apoE−/−) that lacks the primary lipoprotein required for the uptake of lipoproteins through the hepatic receptors, leading to even greater plasma cholesterol increase than in ldlr−/− mice. These and other animal models allowed for conducting genetic studies, such as genome-wide association studies, microarrays, and genotyping methods, which helped identifying more than 100 mutations that contribute to atherosclerosis development. However, translation of the results obtained in animal models for human situations was slow and challenging. At the same time, genetic studies conducted in humans were limited by low sample sizes and high heterogeneity in predictive subclinical phenotypes. In this review, we summarize the current knowledge on the use of KO mice for identification of genes implicated in atherosclerosis and provide a list of genes involved in atherosclerosis-associated inflammatory pathways and their brief characteristics. Moreover, we discuss the approaches for candidate gene search in animals and humans and discuss the progress made in the field of epigenetic studies that appear to be promising for identification of novel biomarkers and therapeutic targets.
Effects of Independent and Combined Water-Deficit and High-Nitrogen Treatments on Flag Leaf Proteomes during Wheat Grain Development
Authors: Zhu Dong , Zhu Gengrui , Zhang Zhen , Wang Zhimin , Yan Xing , Yan Yueming
Published: March 2020

We present the first comprehensive proteome analysis of wheat flag leaves under water-deficit, high-nitrogen (N) fertilization, and combined treatments during grain development in the field. Physiological and agronomic trait analyses showed that leaf relative water content, total chlorophyll content, photosynthetic efficiency, and grain weight and yield were significantly reduced under water-deficit conditions, but dramatically enhanced under high-N fertilization and moderately promoted under the combined treatment. Two-dimensional electrophoresis detected 72 differentially accumulated protein (DAP) spots representing 65 unique proteins, primarily involved in photosynthesis, signal transduction, carbohydrate metabolism, redox homeostasis, stress defense, and energy metabolism. DAPs associated with photosynthesis and protein folding showed significant downregulation and upregulation in response to water-deficit and high-N treatments, respectively. The combined treatment caused a moderate upregulation of DAPs related to photosynthesis and energy and carbohydrate metabolism, suggesting that high-N fertilization can alleviate losses in yield caused by water-deficit conditions by enhancing leaf photosynthesis and grain storage compound synthesis.
Different Trafficking Phenotypes of Niemann-Pick C1 Gene Mutations Correlate with Various Alterations in Lipid Storage, Membrane Composition and Miglustat Amenability
Authors: Brogden Graham , Shammas Hadeel , Walters Friederike , Maalouf Katia , Das Anibh M. , Naim Hassan Y. , Rizk Sandra
Published: March 2020

Niemann-Pick Type C (NPC) is an autosomal recessive lysosomal storage disease leading to progressive neurodegeneration. Mutations in the NPC1 gene, which accounts for 95% of the cases, lead to a defect in intra-lysosomal trafficking of cholesterol and an accumulation of storage material including cholesterol and sphingolipids in the endo-lysosomal system. Symptoms are progressive neurological and visceral deterioration, with variable onset and severity of the disease. This study investigates the influence of two different NPC1 mutations on the biochemical phenotype in fibroblasts isolated from NPC patients in comparison to healthy, wild type (WT) cells. Skin derived fibroblasts were cultured from one patient compound-heterozygous for D874V/D948Y mutations, which presented wild-type like intracellular trafficking of NPC1, and a second patient compound- heterozygous for I1061T/P887L mutations, which exhibited a more severe biochemical phenotype as revealed in the delayed trafficking of NPC1. Biochemical analysis using HPLC and TLC indicated that lipid accumulations were mutation-dependent and correlated with the trafficking pattern of NPC1: higher levels of cholesterol and glycolipids were associated with the mutations that exhibited delayed intracellular trafficking, as compared to their WT-like trafficked or wild type (WT) counterparts. Furthermore, variations in membrane structure was confirmed in these cell lines based on alteration in lipid rafts composition as revealed by the shift in flotillin-2 (FLOT2) distribution, a typical lipid rafts marker, which again showed marked alterations only in the NPC1 mutant showing major trafficking delay. Finally, treatment with N-butyldeoxynojirimycin (NB-DNJ, Miglustat) led to a reduction of stored lipids in cells from both patients to various extents, however, no normalisation in lipid raft structure was achieved. The data presented in this study help in understanding the varying biochemical phenotypes observed in patients harbouring different mutations, which explain why the effectiveness of NB-DNJ treatment is patient specific.
Immunotherapy Strategies for the Prevention and Treatment of Distinct Stages of Type 1 Diabetes: An Overview
Authors: Rapini Novella , Schiaffini Riccardo , Fierabracci Alessandra
Published: March 2020

Type 1 diabetes mellitus is a heterogeneous disorder characterized by destruction of pancreatic β cells, culminating in absolute insulin deficiency. The goals of Type 1 diabetes care, established by the Diabetes Control and Complications Trial (DCCT), are to achieve good glycemic control, to prevent hyperglycaemia (which is associated with long-term microvascular and macrovascular complications) and to avoid recurrent episodes of hypoglycaemia (which may have adverse effects on cognitive function). However, despite continuing optimization of insulin therapy regimes, the actual hormonal substitutive administration acts only to treat the symptoms without an effect on disease pathology and etiopathogenesis. In recent decades, a great deal of interest has been focused on prevention approaches in high-risk individuals, based on the hypothesis that a therapeutic intervention, if applied at the early stage of disease, might contribute to maintaining endogenous β cell function by preserving the residual β cell reservoir from autoimmune attack. This manuscript provides an overview of the most important immunotherapeutic interventions established so far for Type 1 diabetes treatment at different stages of disease that have reached an advanced stage of assessment.
TmPGRP-SA regulates Antimicrobial Response to Bacteria and Fungi in the Fat Body and Gut of Tenebrio molitor
Authors: Keshavarz Maryam , Jo Yong Hun , Edosa Tariku Tesfaye , Bae Young Min , Han Yeon Soo
Published: March 2020

Antimicrobial immune response is mediated by a signal-transducing sensor, peptidoglycan recognition protein-SA (PGRP-SA), that can recognize non-self molecules. Although several studies have focused on the involvement of Drosophila PGRP-SA in antimicrobial peptide (AMP) expression in response to infections, studies on its role in Tenebrio molitor are lacking. Here, we present a functional analysis of T. molitor PGRP-SA (TmPGRP-SA). In the absence of microbes, TmPGRP-SA was highly expressed in the late-larval fat body, followed by hemocytes, and gut. Interestingly, following Escherichia coli, Staphylococcus aureus, and Candida albicans infections, the mRNA level of TmPGRP-SA was significantly upregulated in both the fat body and gut. TmPGRP-SA silencing had a significant effect on the mortality rates for all the microbes tested. Moreover, TmPGRP-SA is required for regulating the expression of eight AMP genes namely TmTenecin-1, -2, and -4; TmDefensin-1 and -2; TmColeoptericin-1; and TmAttacin-1b and -2 in the fat body in response to E. coli and S. aureus infections. TmPGRP-SA is essential for the transcription of TmTenecin-2, -4; TmDefensin-2; TmColeoptericin-1, -2; and TmAttacin-1a, -1b, and -2 in the gut upon E. coli and C. albicans infections. However, TmPGRP-SA does not regulate AMP expression in the hemocytes. Additionally, TmDorsal isoform X2, a downstream Toll transcription factor, was downregulated in TmPGRP-SA-silenced larval fat body following E. coli and S. aureus challenges, and in the gut following E. coli and C. albicans challenges.
Computational Models Using Multiple Machine Learning Algorithms for Predicting Drug Hepatotoxicity with the DILIrank Dataset
Authors: Ancuceanu Robert , Hovanet Marilena Viorica , Anghel Adriana Iuliana , Furtunescu Florentina , Neagu Monica , Constantin Carolina , Dinu Mihaela
Published: March 2020

Drug-induced liver injury (DILI) remains one of the challenges in the safety profile of both authorized and candidate drugs, and predicting hepatotoxicity from the chemical structure of a substance remains a task worth pursuing. Such an approach is coherent with the current tendency for replacing non-clinical tests with in vitro or in silico alternatives. In 2016, a group of researchers from the FDA published an improved annotated list of drugs with respect to their DILI risk, constituting “the largest reference drug list ranked by the risk for developing drug-induced liver injury in humans” (DILIrank). This paper is one of the few attempting to predict liver toxicity using the DILIrank dataset. Molecular descriptors were computed with the Dragon 7.0 software, and a variety of feature selection and machine learning algorithms were implemented in the R computing environment. Nested (double) cross-validation was used to externally validate the models selected. A total of 78 models with reasonable performance were selected and stacked through several approaches, including the building of multiple meta-models. The performance of the stacked models was slightly superior to other models published. The models were applied in a virtual screening exercise on over 100,000 compounds from the ZINC database and about 20% of them were predicted to be non-hepatotoxic.