bluuebunny/crossref_metadata_2025 huggingface dataset:
The Multifunctionality of Exosomes; from the Garbage Bin of the Cell to a Next Generation Gene and Cellular Therapy
Authors: Shrivastava Surya , Morris Kevin V.
Published: January 2021

Exosomes are packaged with a variety of cellular cargo including RNA, DNA, lipids and proteins. For several decades now there has been ongoing debate as to what extent exosomes are the garbage bin of the cell or if these entities function as a distributer of cellular cargo which acts in a meaningful mechanistic way on target cells. Are the contents of exosomes unwanted excess cellular produce or are they selective nucleic acid packaged nanoparticles used to communicate in a paracrine fashion? Overexpressed RNAs and fragments of DNA have been shown to collect into exosomes which are jettisoned from cells in response to particular stimuli to maintain homeostasis suggesting exosomes are functional trash bins of the cell. Other studies however have deciphered selective packaging of particular nucleic acids into exosomes. Nucleic acids packaged into exosomes are increasingly reported to exert transcriptional control on recipient cells, supporting the notion that exosomes may provide a role in signaling and intracellular communication. We survey the literature and conclude that exosomes are multifunctional entities, with a plethora of roles that can each be taken advantage to functionally modulate cells. We also note that the potential utility of developing exosomes as a next generation genetic therapy may in future transform cellular therapies. We also depict three models of methodologies which can be adopted by researchers intending to package nucleic acid in exosomes for developing gene and cell therapy.
Hereditary Haemorrhagic Telangiectasia, an Inherited Vascular Disorder in Need of Improved Evidence-Based Pharmaceutical Interventions
Authors: Snodgrass Ryan O. , Chico Timothy J. A. , Arthur Helen M.
Published: January 2021

Hereditary haemorrhagic telangiectasia (HHT) is characterised by arteriovenous malformations (AVMs). These vascular abnormalities form when arteries and veins directly connect, bypassing the local capillary system. Large AVMs may occur in the lungs, liver and brain, increasing the risk of morbidity and mortality. Smaller AVMs, known as telangiectases, are prevalent on the skin and mucosal lining of the nose, mouth and gastrointestinal tract and are prone to haemorrhage. HHT is primarily associated with a reduction in endoglin (ENG) or ACVRL1 activity due to loss-of-function mutations. ENG and ACVRL1 transmembrane receptors are expressed on endothelial cells (ECs) and bind to circulating ligands BMP9 and BMP10 with high affinity. Ligand binding to the receptor complex leads to activation of the SMAD1/5/8 signalling pathway to regulate downstream gene expression. Various genetic animal models demonstrate that disruption of this pathway in ECs results in AVMs. The vascular abnormalities underlying AVM formation result from abnormal EC responses to angiogenic and haemodynamic cues, and include increased proliferation, reduced migration against the direction of blood flow and an increased EC footprint. There is growing evidence that targeting VEGF signalling has beneficial outcomes in HHT patients and in animal models of this disease. The anti-VEGF inhibitor bevacizumab reduces epistaxis and has a normalising effect on high cardiac output in HHT patients with hepatic AVMs. Blocking VEGF signalling also reduces vascular malformations in mouse models of HHT1 and HHT2. However, VEGF signalling is complex and drives numerous downstream pathways, and it is not yet clear which pathway (or combination of pathways) is critical to target. This review will consider the recent evidence gained from HHT clinical and preclinical studies that are increasing our understanding of HHT pathobiology and informing therapeutic strategies.
Long Non-Coding RNA FENDRR: Gene Structure, Expression, and Biological Relevance
Authors: Szafranski Przemyslaw , Stankiewicz Paweł
Published: January 2021

The FOXF1 Adjacent Noncoding Developmental Regulatory RNA (Fendrr) plays an important role in the control of gene expression in mammals. It is transcribed in the opposite direction to the neighboring Foxf1 gene with which it shares a region containing promoters. In humans, FENDRR is located on chromosome 16q24.1, and is positively regulated both by the FOXF1 distant lung-specific cis-acting enhancer and by trans-acting FOXF1. Fendrr has been shown to function as a competing endogenous RNA, sponging microRNAs and protein factors that control stability of mRNAs, and as an epigenetic modifier of chromatin structure around gene promoters and other regulatory sites, targeting them with histone methyltrasferase complexes. In mice, Fendrr is essential for development of the heart, lungs, and gastrointestinal system; its homozygous loss causes embryonic or perinatal lethality. Importantly, deregulation of FENDRR expression has been causatively linked also to tumorigenesis, resistance to chemotherapy, fibrosis, and inflammatory diseases. Here, we review the current knowledge on the FENDRR structure, expression, and involvement in development and tissue maintenance.
Integrated Analysis of Methylomic and Transcriptomic Data to Identify Potential Diagnostic Biomarkers for Major Depressive Disorder
Authors: Xie Yinping , Xiao Ling , Chen Lijuan , Zheng Yage , Zhang Caixia , Wang Gaohua
Published: January 2021

Major depressive disorder (MDD) is a mental illness with high incidence and complex etiology, that poses a serious threat to human health and increases the socioeconomic burden. Currently, high-accuracy biomarkers for MDD diagnosis are urgently needed. This paper aims to identify novel blood-based diagnostic biomarkers for MDD. Whole blood DNA methylation data and gene expression data from the Gene Expression Omnibus database are downloaded. Then, differentially expressed/methylated genes (DEGs/DMGs) are identified. In addition, we made a systematic analysis of the DNA methylation on 5′-C-phosphate-G-3′ (CpGs) in all of the gene regions, as well as different gene regions, and then we defined a “dominant” region. Subsequently, integrated analysis is employed to identify the robust MDD-related blood biomarkers. Finally, a gene expression classifier and a methylation classifier are constructed using the random forest algorithm and the leave-one-out cross-validation method. Our results demonstrate that DEGs are mainly involved in the inflammatory response-associated pathways, while DMGs are primarily concentrated in the neurodevelopment- and neuroplasticity-associated pathways. Our integrated analysis identified 46 hypo-methylated and up-regulated (hypo-up) genes and 71 hyper-methylated and down-regulated (hyper-down) genes. One gene expression classifier and two DNA methylation classifiers, based on the CpGs in all of the regions or in the dominant regions are constructed. The gene expression classifier possessed the best predictive ability, followed by the DNA methylation classifiers, based on the CpGs in both the dominant regions and all of the regions. In summary, the integrated analysis of DNA methylation and gene expression has identified 46 hypo-up genes and 71 hyper-down genes, which could be used as diagnostic biomarkers for MDD.
Butanol Tolerance of Lactiplantibacillus plantarum: A Transcriptome Study
Authors: Petrov Kaloyan , Arsov Alexander , Petrova Penka
Published: January 2021

Biobutanol is a promising alternative fuel with impaired microbial production thanks to its toxicity. Lactiplantibacillus plantarum (L. plantarum) is among the few bacterial species that can naturally tolerate 3% (v/v) butanol. This study aims to identify the genetic factors involved in the butanol stress response of L. plantarum by comparing the differential gene expression in two strains with very different butanol tolerance: the highly resistant Ym1, and the relatively sensitive 8-1. During butanol stress, a total of 319 differentially expressed genes (DEGs) were found in Ym1, and 516 in 8-1. Fifty genes were upregulated and 54 were downregulated in both strains, revealing the common species-specific effects of butanol stress: upregulation of multidrug efflux transporters (SMR, MSF), toxin-antitoxin system, transcriptional regulators (TetR/AcrR, Crp/Fnr, and DeoR/GlpR), Hsp20, and genes involved in polysaccharide biosynthesis. Strong inhibition of the pyrimidine biosynthesis occurred in both strains. However, the strains differed greatly in DEGs responsible for the membrane transport, tryptophan synthesis, glycerol metabolism, tRNAs, and some important transcriptional regulators (Spx, LacI). Uniquely upregulated in the butanol-resistant strain Ym1 were the genes encoding GntR, GroEL, GroES, and foldase PrsA. The phosphoenolpyruvate flux and the phosphotransferase system (PTS) also appear to be major factors in butanol tolerance.
Graph Algorithms for Mixture Interpretation
Authors: Crysup Benjamin , Woerner August E. , King Jonathan L. , Budowle Bruce
Published: January 2021

The scale of genetic methods are presently being expanded: forensic genetic assays previously were limited to tens of loci, but now technologies allow for a transition to forensic genomic approaches that assess thousands to millions of loci. However, there are subtle distinctions between genetic assays and their genomic counterparts (especially in the context of forensics). For instance, forensic genetic approaches tend to describe a locus as a haplotype, be it a microhaplotype or a short tandem repeat with its accompanying flanking information. In contrast, genomic assays tend to provide not haplotypes but sequence variants or differences, variants which in turn describe how the alleles apparently differ from the reference sequence. By the given construction, mitochondrial genetic assays can be thought of as genomic as they often describe genetic differences in a similar way. The mitochondrial genetics literature makes clear that sequence differences, unlike the haplotypes they encode, are not comparable to each other. Different alignment algorithms and different variant calling conventions may cause the same haplotype to be encoded in multiple ways. This ambiguity can affect evidence and reference profile comparisons as well as how “match” statistics are computed. In this study, a graph algorithm is described (and implemented in the MMDIT (Mitochondrial Mixture Database and Interpretation Tool) R package) that permits the assessment of forensic match statistics on mitochondrial DNA mixtures in a way that is invariant to both the variant calling conventions followed and the alignment parameters considered. The algorithm described, given a few modest constraints, can be used to compute the “random man not excluded” statistic or the likelihood ratio. The performance of the approach is assessed in in silico mitochondrial DNA mixtures.
VIRMOTIF: A User-Friendly Tool for Viral Sequence Analysis
Authors: Rajaei Pedram , Jahanian Khadijeh Hoda , Beheshti Amin , Band Shahab S. , Dehzangi Abdollah , Alinejad-Rokny Hamid
Published: January 2021

Bioinformatics and computational biology have significantly contributed to the generation of vast and important knowledge that can lead to great improvements and advancements in biology and its related fields. Over the past three decades, a wide range of tools and methods have been developed and proposed to enhance performance, diagnosis, and throughput while maintaining feasibility and convenience for users. Here, we propose a new user-friendly comprehensive tool called VIRMOTIF to analyze DNA sequences. VIRMOTIF brings different tools together as one package so that users can perform their analysis as a whole and in one place. VIRMOTIF is able to complete different tasks, including computing the number or probability of motifs appearing in DNA sequences, visualizing data using the matplotlib and heatmap libraries, and clustering data using four different methods, namely K-means, PCA, Mean Shift, and ClusterMap. VIRMOTIF is the only tool with the ability to analyze genomic motifs based on their frequency and representation (D-ratio) in a virus genome.
Validity and Psychometric Properties of the Internet Gaming Disorder Scale in Three Large Independent Samples of Children and Adolescents
Authors: Paschke Kerstin , Sack Peter-Michael , Thomasius Rainer
Published: January 2021

Background: Problematic gaming has become a major health issue in children and adolescents resulting in the need for targeted valid and reliable screening instruments. This study aimed to explore the psychometric properties and criterion validity of the widely used 9-item Internet Gaming Disorder Scale (IGDS) in young gamers. Methods: Three independent samples were drawn from socio-demographically representative cross-sectional telephone surveys collected in the years 2016 (N = 762), 2017 (N = 777), and 2018 (N = 784) and analyzed separately. Results: The IGDS revealed psychometric properties suitable for screening in large samples. Cronbach’s alpha was 0.563, 0.724, and 0.778. The unidimensionality assumption was challenged. At-risk and pathological gamers compared to normal gamers reported longer digital media use and more emotional symptoms and hyperactivity/inattention with clinical relevance to medium effect sizes. The comparison of at-risk and pathological gamers indicated a partial distinction between the two problematic gaming groups. Conclusions: The IGDS could be shown to be an overall suitable and valid tool to identify pathological gamers in childhood and adolescence according to the DSM-5 criteria on a population level. However, the polythetic structure limits comparability with the recent ICD-11 criteria. At-risk gamers appeared as a heterogeneous group warranting more research.
Regional Disparity and Patients Mobility: Benefits and Spillover Effects of the Spatial Network Structure of the Health Services in China
Authors: Fu Liping , Xu Kaibo , Liu Feng , Liang Lu , Wang Zhengmin
Published: January 2021

Background: The distribution of medical resources in China is seriously imbalanced due to imbalanced economic development in the country; unbalanced distribution of medical resources makes patients try to seek better health services. Against this backdrop, this study aims to analyze the spatial network characteristics and spatial effects of China’s health economy, and then find evidence that affects patient mobility. Methods: Data for this study were drawn from the China Health Statistical Yearbooks and China Statistical Books. The gravitational value of China’s health spatial network was calculated to establish a network of gravitational relationships. The social network analysis method was used for centrality analysis and spillover effect analysis. Results: A gravity correlation matrix was constructed among provinces by calculating the gravitational value, indicating the spatial relationships of different provinces in the health economic network. Economically developed provinces, such as Shanghai and Jiangsu, are at the center of the health economic network (centrality degree = 93.333). These provinces also play a strong intermediary role in the network and have connections with other provinces. In the CONCOR analysis, 31 provinces are divided into four blocks. The spillover effect of the blocks indicates provinces with medical resource centers have beneficial effects, while provinces with insufficient resources have obvious spillover effects. Conclusion: There is a significant gap in the geographical distribution of medical resources, and the health economic spatial network structure needs to be improved. Most medical resources are concentrated in economically developed provinces, and these provinces’ positions in the health economic spatial network are becoming more centralized. By contrast, economically underdeveloped regions are at the edge of the network, causing patients to move to provinces with medical resource centers. There are health risks of the increasing pressure to seek medical treatment in developed provinces with abundant medical resources.
Physical Activity, Mental Health and Consumption of Medications in Pre-Elderly People: The National Health Survey 2017
Authors: Carmona-Torres Juan Manuel , Cobo-Cuenca Ana Isabel , Pozuelo-Carrascosa Diana P. , Latorre-Román Pedro Ángel , Párraga-Montilla Juan Antonio , Laredo-Aguilera José Alberto
Published: January 2021

Background: The promotion of Physical Activity (PA) is an important public health goal to reduce comorbidity and diseases associated with aging such as anxiety and depression. Aim: To investigate the association between level of PA, mental health and the consumption of medications among a representative cohort of Spanish pre-elderly people. Methods: Cross-sectional study with 5977 participants aged 50 to 64 years who participated in the National Health Survey in Spain 2017. The levels of PA were evaluated using the International Physical Activity Questionnaire and the mental conditions were measured by the Goldberg Health Questionnaire-12 (GHQ-12). The chi-square test was used for qualitative variables; Pearson’s correlation was conducted between GHQ-12 score with different quantitative variables; and a logistic regression was used to determine the association between PA and mental health with the sociodemographic characteristics. Results: The participants were 51.9% women and 48.1% men with a mean age of 56.79 years, and 35.5% of participants had a low level of PA. A low level of PA was associated with cases of mental health vulnerability, anxiety and depression (in women), the consumption of more medications and greater multimorbidity. Conclusion: It is important that people reach old age with an optimal health status in order to reduce age-related disability and morbidity. More than a third of the Spanish pre-elderly do not reach the levels of PA recommended by the WHO. People who had low level of PA consumed more medications and had higher mental health vulnerability and greater multimorbidity.
Sinus Floor Elevation via an Osteotome Technique without Biomaterials
Authors: Andrés-García Rodrigo , Ríos-Santos José Vicente , Herrero-Climent Mariano , Bullón Pedro , Fernández-Farhall Javier , Gómez-Menchero Alberto , Fernández-Palacín Ana , Ríos-Carrasco Blanca
Published: January 2021

According to classic Hirschfeld studies, the first teeth to be lost are the first and second maxillary molars. After the teeth are extracted and the alveolar process is developed, the maxillary sinus is reabsorbed and pneumatized with a decrease in bone availability in the posterior sector of the maxilla. This process often creates the need to perform regeneration techniques for the placement of implants in this area due to the low availability of bone. The most frequently used and documented technique for the elevation of the sinus maxillary floor is elevation by the side window, as proposed by Tatum. In 1994, Summers proposed a technique that allowed the elevation of the sinus floor from a crestal access using an instrument called an osteotome, as well as the placement of the implant in the same surgical act. The aimed of the study was to evaluate the survival of 32 implants placed in posterior maxilla with bone availability less than 5 mm performing a sinus lift augmentation technique with osteotome without biomaterials. The results of this study show a survival rate of 100% for 32 implants placed in situations with an initial bone availability of 2 to 5 mm without the use of graft material. The infra-drilling technique used offers an increase in the primary stability of implants that allows adequate osteointegration Implants placed were charged at 12 weeks. In all cases, spontaneous bone formation was observed, even in cases where a positive Valsalva maneuver was observed. This proposed technique reduces treatment time and the need for more invasive maxillary sinus augmentation techniques.
COVID-19-Related Changes in Perceived Household Food Waste in the United States: A Cross-Sectional Descriptive Study
Authors: Cosgrove Kelly , Vizcaino Maricarmen , Wharton Christopher
Published: January 2021

Food waste contributes to adverse environmental and economic outcomes, and substantial food waste occurs at the household level in the US. This study explored perceived household food waste changes during the COVID-19 pandemic and related factors. A total of 946 survey responses from primary household food purchasers were analyzed. Demographic, COVID-19-related household change, and household food waste data were collected in October 2020. Wilcoxon signed-rank was used to assess differences in perceived food waste. A hierarchical binomial logistic regression analysis was conducted to examine whether COVID-19-related lifestyle disruptions and food-related behavior changes increased the likelihood of household food waste. A binomial logistic regression was conducted to explore the contribution of different food groups to the likelihood of increased food waste. Perceived food waste, assessed as the estimated percent of food wasted, decreased significantly during the pandemic (z = −7.47, p < 0.001). Food stockpiling was identified as a predictor of increased overall food waste during the pandemic, and wasting fresh vegetables and frozen foods increased the odds of increased food waste. The results indicate the need to provide education and resources related to food stockpiling and the management of specific food groups during periods of disruption to reduce food waste.
A Spatio-Temporal Analysis of the Health Situation in Poland Based on Functional Discriminant Coordinates
Authors: Krzyśko Mirosław , Wołyńki Waldemar , Szymkowiak Marcin , Wojtyła Andrzej
Published: January 2021

The aim of this study was to investigate if the provinces of Poland are homogeneous in terms of the observed spatio-temporal data characterizing the health situation of their inhabitants. The health situation is understood as a set of selected factors influencing inhabitants’ health and the healthcare system in their area of residence. So far, studies concerning the health situation of selected territorial units have been based on data relating to a specific year rather than longer periods. The task of assessing province homogeneity was carried out in two stages. In stage one, the original spatio-temporal data space (space of multivariate time series) was transformed into a functional discriminant coordinates space. The resulting functional discriminant coordinates are synthetic measures of the health situation of inhabitants of particular provinces. These measures contain complete information regarding 8 diagnostic variables examined over a period of 6 years. In the second stage, the Ward method, commonly used in cluster analysis, was applied in order to identify groups of homogeneous provinces in the space of functional discriminant coordinates. Sixteen provinces were divided into four clusters. The homogeneity of the clusters was confirmed by the multivariate functional coefficient of variation.
Rural Financial Development Impacts on Agricultural Technology Innovation: Evidence from China
Authors: Liu Yuyu , Ji Duan , Zhang Lin , An Jingjing , Sun Wenyan
Published: January 2021

Agricultural technology innovation is key for improving productivity, sustainability, and resilience in food production and agriculture to contribute to public health. Using panel data of 31 provinces in China from 2003 to 2015, this study examines the impact of rural financial development on agricultural technology innovation from the perspective of rural financial scale and rural finance efficiency. Furthermore, it examines how the effects of rural financial development vary in regions with different levels of marketization and economic development. The empirical results show that the development of rural finance has a significant and positive effect on the level of agricultural technology innovation. Rural finance efficiency has a significantly positive effect on innovation in regions with a low degree of marketization, while the rural financial scale has a significantly positive effect on technological innovation in regions with a high degree of marketization. Further analysis showed that improving the level of agricultural technology innovation is conducive to rural economic development. This study provides new insights into the effects of rural financial development on sustainable agricultural development from the perspective of agricultural technology innovation.
Agriculture Development, Pesticide Application and Its Impact on the Environment
Authors: Tudi Muyesaier , Daniel Ruan Huada , Wang Li , Lyu Jia , Sadler Ross , Connell Des , Chu Cordia , Phung Dung Tri
Published: January 2021

Pesticides are indispensable in agricultural production. They have been used by farmers to control weeds and insects, and their remarkable increases in agricultural products have been reported. The increase in the world’s population in the 20th century could not have been possible without a parallel increase in food production. About one-third of agricultural products are produced depending on the application of pesticides. Without the use of pesticides, there would be a 78% loss of fruit production, a 54% loss of vegetable production, and a 32% loss of cereal production. Therefore, pesticides play a critical role in reducing diseases and increasing crop yields worldwide. Thus, it is essential to discuss the agricultural development process; the historical perspective, types and specific uses of pesticides; and pesticide behavior, its contamination, and adverse effects on the natural environment. The review study indicates that agricultural development has a long history in many places around the world. The history of pesticide use can be divided into three periods of time. Pesticides are classified by different classification terms such as chemical classes, functional groups, modes of action, and toxicity. Pesticides are used to kill pests and control weeds using chemical ingredients; hence, they can also be toxic to other organisms, including birds, fish, beneficial insects, and non-target plants, as well as air, water, soil, and crops. Moreover, pesticide contamination moves away from the target plants, resulting in environmental pollution. Such chemical residues impact human health through environmental and food contamination. In addition, climate change-related factors also impact on pesticide application and result in increased pesticide usage and pesticide pollution. Therefore, this review will provide the scientific information necessary for pesticide application and management in the future.
Serum Concentration of Fluoride in Patients with Alcoholic Liver Cirrhosis from the Lublin Region in Eastern Poland
Authors: Prystupa Andrzej , Sak Jarosław , Kiciński Paweł , Stenzel-Bembenek Agnieszka , Błażewicz Anna
Published: January 2021

In view of previous reports, it is important to determine the relationship between liver function and the level of fluoride in the serum. The aim of this study was to investigate serum concentrations of fluoride in 72 patients with alcoholic liver cirrhosis, living in the region of Lublin (Eastern Poland) divided based on the severity of disease according to the Child-Turcotte-Pugh criteria. Higher plasma fluoride concentrations were associated with changes in liver related parameters. In all groups of analyzed patients with different stages of alcoholic liver cirrhosis, elevated levels of plasma fluoride and increased activities of both alanine aminotransferase (ALT) and total bilirubin concentration were shown.
Spatial Patterns of COVID-19 Incidence in Relation to Crime Rate Across London
Authors: Sun Yeran , Huang Ying , Yuan Ke , Chan Ting On , Wang Yu
Published: January 2021

COVID-19 containment policies are not only curbing the spread of COVID-19 but also changing human behavior. According to the routine activity theory, owing to lockdown, the closure of entertainment sites (e.g., pubs and bars), an increase in stay-at-home time, and an increase in police patrols are likely to influence chance of committing a crime. In this study, we aimed to further examine the spatial association of COVID-19 infection rate and crime rate. Particularly, we empirically validated the speculation that increase in COVID-19 cases is likely to reduce crime rate. In the empirical study, we investigated whether and how COVID-19 infection rate is spatially associated with crime rate in London. As the spatial data used are mainly areal data, we adopted a spatial regression mode (i.e., the “random effects eigenvector spatial filtering model”) to investigate the spatial associations after controlling for the socioeconomic factors. More specifically, we investigated the associations for all the four crime categories in three consequent months (March, April, and May of 2020). The empirical results indicate that 1) crime rates of the four categories have no statistically significant associations with COVID-19 infection rate in March; 2) violence-against-the-person rate has no statistically significant association with COVID-19 infection rate; and 3) robbery rate, burglary rate, and theft and handling rate have a statistically significant and negative association with COVID-19 infection rate in both April and May.
Phytochemicals in Gynecological Cancer Prevention
Authors: Woźniak Marta , Krajewski Rafał , Makuch Sebastian , Agrawal Siddarth
Published: January 2021

Gynecological cancer confers an enormous burden among women worldwide. Accumulating evidence points to the role of phytochemicals in preventing cervical, endometrial, and ovarian cancer. Experimental studies emphasize the chemopreventive and therapeutic potential of plant-derived substances by inhibiting the early stages of carcinogenesis or improving the efficacy of traditional chemotherapeutic agents. Moreover, a number of epidemiological studies have investigated associations between a plant-based diet and cancer risk. This literature review summarizes the current knowledge on the phytochemicals with proven antitumor activity, emphasizing their effectiveness and mechanism of action in gynecological cancer.
Coxsackievirus B3 Infection of Human iPSC Lines and Derived Primary Germ-Layer Cells Regarding Receptor Expression
Authors: Böhnke Janik , Pinkert Sandra , Schmidt Maria , Binder Hans , Bilz Nicole Christin , Jung Matthias , Reibetanz Uta , Beling Antje , Rujescu Dan , Claus Claudia
Published: January 2021

The association of members of the enterovirus family with pregnancy complications up to miscarriages is under discussion. Here, infection of two different human induced pluripotent stem cell (iPSC) lines and iPSC-derived primary germ-layer cells with coxsackievirus B3 (CVB3) was characterized as an in vitro cell culture model for very early human development. Transcriptomic analysis of iPSC lines infected with recombinant CVB3 expressing enhanced green fluorescent protein (EGFP) revealed a reduction in the expression of pluripotency genes besides an enhancement of genes involved in RNA metabolism. The initial distribution of CVB3-EGFP-positive cells within iPSC colonies correlated with the distribution of its receptor coxsackie- and adenovirus receptor (CAR). Application of anti-CAR blocking antibodies supported the requirement of CAR, but not of the co-receptor decay-accelerating factor (DAF) for infection of iPSC lines. Among iPSC-derived germ-layer cells, mesodermal cells were especially vulnerable to CVB3-EGFP infection. Our data implicate further consideration of members of the enterovirus family in the screening program of human pregnancies. Furthermore, iPSCs with their differentiation capacity into cell populations of relevant viral target organs could offer a reliable screening approach for therapeutic intervention and for assessment of organ-specific enterovirus virulence.
Central Apolipoprotein A-IV Stimulates Thermogenesis in Brown Adipose Tissue
Authors: Pence Sydney , LaRussa Zachary , Shen Zhijun , Liu Min , Coschigano Karen , Shi Haifei , Lo Chunmin
Published: January 2021

Stimulation of thermogenesis in brown adipose tissue (BAT) could have far-reaching health benefits in combatting obesity and obesity-related complications. Apolipoprotein A-IV (ApoA-IV), produced by the gut and the brain in the presence of dietary lipids, is a well-known short-term satiating protein. While our previous studies have demonstrated reduced diet-induced thermogenesis in ApoA-IV-deficient mice, it is unclear whether this reduction is due to a loss of peripheral or central effects of ApoA-IV. We hypothesized that central administration of ApoA-IV stimulates BAT thermogenesis and that sympathetic and sensory innervation is necessary for this action. To test this hypothesis, mice with unilateral denervation of interscapular BAT received central injections of recombinant ApoA-IV protein or artificial cerebrospinal fluid (CSF). The effects of central ApoA-IV on BAT temperature and thermogenesis in mice with unilateral denervation of the intrascapular BAT were monitored using transponder probe implantation, qPCR, and immunoblots. Relative to CSF, central administration of ApoA-IV significantly increased temperature and UCP expression in BAT. However, all of these effects were significantly attenuated or prevented in mice with unilateral denervation. Together, these results clearly demonstrate that ApoA-IV regulates BAT thermogenesis centrally, and this effect is mediated through sympathetic and sensory nerves.
Blood-Brain Barrier Disruption Increases Amyloid-Related Pathology in TgSwDI Mice
Authors: Abdallah Ihab M. , Al-Shami Kamal M. , Yang Euitaek , Kaddoumi Amal
Published: January 2021

In Alzheimer’s disease (AD), several studies have reported blood-brain barrier (BBB) breakdown with compromised function. P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) are transport proteins localized at the BBB luminal membrane and play an important role in the clearance of amyloid-β (Aβ). The purpose of this study was to investigate the effect of pharmacological inhibition of Aβ efflux transporters on BBB function and Aβ accumulation and related pathology. Recently, we have developed an in vitro high-throughput screening assay to screen for compounds that modulate the integrity of a cell-based BBB model, which identified elacridar as a disruptor of the monolayer integrity. Elacridar, an investigational compound known for its P-gp and BCRP inhibitory effect and widely used in cancer research. Therefore, it was used as a model compound for further evaluation in a mouse model of AD, namely TgSwDI. TgSwDI mouse is also used as a model for cerebral amyloid angiopathy (CAA). Results showed that P-gp and BCRP inhibition by elacridar disrupted the BBB integrity as measured by increased IgG extravasation and reduced expression of tight junction proteins, increased amyloid deposition due to P-gp, and BCRP downregulation and receptor for advanced glycation end products (RAGE) upregulation, increased CAA and astrogliosis. Further studies revealed the effect was mediated by activation of NF-κB pathway. In conclusion, results suggest that BBB disruption by inhibiting P-gp and BCRP exacerbates AD pathology in a mouse model of AD, and indicate that therapeutic drugs that inhibit P-gp and BCRP could increase the risk for AD.
Filifactor alocis and Tumor Necrosis Factor-Alpha Stimulate Synthesis of Visfatin by Human Macrophages
Authors: Nogueira Andressa Vilas Boas , Nokhbehsaim Marjan , Damanaki Anna , Eick Sigrun , Kirschneck Christian , Schröder Agnes , Jantsch Jonathan , Deschner James
Published: January 2021

There is little known about the effect of the periodontopathogen Filifactor alocis on macrophages as key cells of the innate immune defense in the periodontium. Therefore, the aim of the present study was to investigate the effect of F. alocis and additionally of the pro-inflammatory cytokine tumor necrosis factor-alpha (TNFα) on visfatin and other pro-inflammatory and proteolytic molecules associated with periodontitis in human macrophages. The presence of macrophage markers CD14, CD86, CD68, and CD163 was examined in gingival biopsies from healthy individuals and periodontitis patients. Human macrophages were incubated with F. alocis and TNFα for up to 2 d. The effects of both stimulants on macrophages were determined by real-time PCR, ELISA, immunocytochemistry, and immunofluorescence. F. alocis was able to significantly stimulate the synthesis of visfatin by human macrophages using TLR2 and MAPK pathways. In addition to visfatin, F. alocis was also able to increase the synthesis of cyclooxygenase 2, TNFα, and matrix metalloproteinase 1. Like F. alocis, TNFα was also able to stimulate the production of these proinflammatory and proteolytic molecules. Our results highlight the pathogenetic role of F. alocis in periodontal diseases and also underline the involvement of visfatin in the aetiopathogenesis of periodontitis.
Design of RGDS Peptide-Immobilized Self-Assembling β-Strand Peptide from Barnacle Protein
Authors: Fujii Daisuke , Takase Kento , Takagi Ami , Kamino Kei , Hirano Yoshiaki
Published: January 2021

We designed three types of RGD-containing barnacle adhesive proteins using self-assembling peptides. In the present study, three types of RGD-containing peptides were synthesized by solid-phase peptide synthesis, and the secondary structures of these peptides were analyzed by CD and FT-IR spectroscopy. The mechanical properties of peptide hydrogels were characterized by a rheometer. We discuss the correlation between the peptide conformation, and cell attachment and cell spreading activity from the viewpoint of developing effective tissue engineering scaffolds. We created a peptide-coated cell culture substrate by coating peptides on a polystyrene plate. They significantly facilitated cell adhesion and spreading compared to a non-coated substrate. When the RGDS sequence was modified at N- or C-terminal of R-Y, it was found that the self-assembling ability was dependent on the strongly affects hydrogel formation and cell adhesion caused by its secondary structure.
Centrifugation Removes a Population of Large Vesicles, or “Macroparticles,” Intermediate in Size to RBCs and Microvesicles
Authors: Larson Michael C. , Hogg Neil , Hillery Cheryl A.
Published: January 2021

Microparticles or microvesicles (MPs/MVs) are sub-cellular vesicles with a growing number of known biological functions. Microvesicles from a variety of parent cells within the vascular system increase in numerous pathological states. Red blood cell-derived MVs (RMVs) are relatively less studied than other types of circulating MVs despite red blood cells (RBCs) being the most abundant intravascular cell. This may be in part due the echoes of past misconceptions that RBCs were merely floating anucleate bags of hemoglobin rather than dynamic and responsive cells. The initial aim of this study was to maximize the concentration of RMVs derived from various blood or blood products by focusing on the optimal isolation conditions without creating more MVs from artificial manipulation. We found that allowing RBCs to sediment overnight resulted in a continuum in size of RBC membrane-containing fragments or vesicles extending beyond the 1 µm size limit suggested by many as the maximal size of an MV. Additionally, dilution and centrifugation factors were studied that altered the resultant MV population concentration. The heterogeneous size of RMVs was confirmed in mice models of hemolytic anemia. This methodological finding establishes a new paradigm in that it blurs the line between RBC, fragment, and RMV as well as suggests that the concentration of circulating RMVs may be widely underestimated given that centrifugation removes the majority of such RBC-derived membrane-containing particles.
HIF1A: A Putative Modifier of Hemochromatosis
Authors: Pelucchi Sara , Ravasi Giulia , Arosio Cristina , Mauri Mario , Piazza Rocco , Mariani Raffaella , Piperno Alberto
Published: January 2021

HFE-related hereditary hemochromatosis (HH) is characterized by marked phenotypic heterogeneity. Homozygosity for p.C282Y is a low penetrance genotype suggesting that the HFE-HH is a multifactorial disease resulting from a complex interaction involving a major gene defect, genetic background and environmental factors. We performed a targeted NGS-based gene panel to identify new candidate modifiers by using an extreme phenotype sampling study based on serum ferritin and iron removed/age ratio. We found an increased prevalence of the HIF1A p.Phe582Ser and p.Ala588Thr variants in patients with a severe iron and clinical phenotype. Accordingly, Huh-7 cells transfected with both variants showed significantly lower HAMP promoter activity by luciferase assay. The qRT-PCR assays showed a downregulation of hepcidin and an upregulation of the HIF1A target genes (VEGF, HMOX, FUR, TMPRSS6) in cells transfected with the HIF1A-P582S vector. We identified mutations in other genes (e.g., Serpina1) that might have some relevance in single cases in aggravating or mitigating disease manifestation. In conclusion, the present study identified HIF1A as a possible modifier of the HFE-HH phenotype cooperating with the genetic defect in downregulating hepcidin synthesis. In addition, this study highlights that an NGS-based approach could broaden our knowledge and help in characterizing the genetic complexity of HFE-HH patients with a severe phenotype expression.
Integrating LCM-Based Spatio-Temporal Transcriptomics Uncovers Conceptus and Endometrial Luminal Epithelium Communication that Coordinates the Conceptus Attachment in Pigs
Authors: Wang Feiyu , Zhao Shilei , Deng Dadong , Wang Weiwei , Xu Xuewen , Liu Xiaolei , Zhao Shuhong , Yu Mei
Published: January 2021

Attachment of conceptus to the endometrial luminal epithelium (LE) is a critical event for early placentation in Eutheria. Since the attachment occurs at a particular site within the uterus, a coordinated communication between three spatially distinct compartments (conceptus and endometrial LE from two anatomical regions of the uterus to which conceptus attaches and does not attach) is essential but remains to be fully characterized. Using the laser capture microdissection (LCM) technique, we firstly developed an approach that can allow us to pair the pig conceptus sample with its nearby endometrial epithelium sample without losing the native spatial information. Then, a comprehensive spatio-temporal transcriptomic profile without losing the original conceptus-endometrium coordinates was constructed. The analysis shows that an apparent difference in transcriptional responses to the conceptus exists between the endometrial LE from the two anatomically distinct regions in the uterus. In addition, we identified the communication pathways that link the conceptus and endometrial LE and found that these pathways have important roles in conceptus attachment. Furthermore, a number of genes whose expression is spatially restricted in the two different anatomical regions within the uterus were characterized for the first time and two of them (SULT2A1 and MEP1B) may cooperatively contribute to establish conceptus attachment in pigs. The results from our study have implications in understanding of conceptus/embryo attachment in pigs and other large polytocous species.
Effect of Pd Precursor Salts on the Chemical State, Particle Size, and Performance of Activated Carbon-Supported Pd Catalysts for the Selective Hydrogenation of Palm Biodiesel
Authors: Udomsap Parncheewa , Eiad-Ua Apiluck , Chen Shih-Yuan , Mochizuki Takehisa , Chollacoop Nuwong , Yoshimura Yuji , Nishi Masayasu , Tateno Hiroyuki , Takagi Hideyuki
Published: January 2021

To improve the oxidative stability of biodiesel fuel (BDF), the polyunsaturated fatty acid methyl esters (poly-FAME) presented in commercial palm oil-derived biodiesel fuel (palm-BDF) were selectively hydrogenated to monounsaturated fatty acid methyl esters (mono-FAME) under a mild condition (80 °C, 0.5 MPa) using activated carbon (AC)-supported Pd catalysts with a Pd loading of 1 wt.%. The partially hydrotreated palm-BDF (denoted as H-FAME) which has low poly-FAME components is a new type of BDF with enhanced quality for use in high blends. In this study, we reported that the chemical states and particle sizes of Pd in the prepared Pd/AC catalysts were significantly influenced by the Pd precursors, Pd(NO3)2 and Pd(NH3)4Cl2, and thus varied their hydrogenation activity and product selectivity. The 1%Pd/AC (nit) catalyst, prepared using Pd(NO3)2, presented high performance for selective hydrogenation of poly-FAME into mono-FAME with high oxidation stability, owning to its large Pd particles (8.4 nm). Conversely, the 1%Pd/AC (amc) catalyst, prepared using Pd(NH3)4Cl2, contained small Pd particles (2.7 nm) with a little Cl residues, which could be completely removed by washing with an aqueous solution of 0.1 M NH4OH. The small Pd particles gave increased selectivity toward unwanted-FAME components, particularly the saturated fatty acid methyl esters during the hydrogenation of poly-FAME. This selectivity is unprofitable for improving the biodiesel quality.
Neuroprotective Effect of Apolipoprotein D in Cuprizone-Induced Cell Line Models: A Potential Therapeutic Approach for Multiple Sclerosis and Demyelinating Diseases
Authors: Martínez-Pinilla Eva , Rubio-Sardón Núria , Peláez Rafael , García-Álvarez Enrique , del Valle Eva , Tolivia Jorge , Larráyoz Ignacio M. , Navarro Ana
Published: January 2021

Apolipoprotein D (Apo D) overexpression is a general finding across neurodegenerative conditions so the role of this apolipoprotein in various neuropathologies such as multiple sclerosis (MS) has aroused a great interest in last years. However, its mode of action, as a promising compound for the development of neuroprotective drugs, is unknown. The aim of this work was to address the potential of Apo D to prevent the action of cuprizone (CPZ), a toxin widely used for developing MS models, in oligodendroglial and neuroblastoma cell lines. On one hand, immunocytochemical quantifications and gene expression measures showed that CPZ compromised neural mitochondrial metabolism but did not induce the expression of Apo D, except in extremely high doses in neurons. On the other hand, assays of neuroprotection demonstrated that antipsychotic drug, clozapine, induced an increase in Apo D synthesis only in the presence of CPZ, at the same time that prevented the loss of viability caused by the toxin. The effect of the exogenous addition of human Apo D, once internalized, was also able to directly revert the loss of cell viability caused by treatment with CPZ by a reactive oxygen species (ROS)-independent mechanism of action. Taken together, our results suggest that increasing Apo D levels, in an endo- or exogenous way, moderately prevents the neurotoxic effect of CPZ in a cell model that seems to replicate some features of MS which would open new avenues in the development of interventions to afford MS-related neuroprotection.
Five Silkworm 30K Proteins Are Involved in the Cellular Immunity against Fungi
Authors: Ye Lin , Zhang Yan , Dong Zhaoming , Guo Pengchao , Zhao Dongchao , Li Haoyun , Hu Hang , Zhou Xiaofang , Chen Haiqin , Zhao Ping
Published: January 2021

Background: 30K proteins are a major group of nutrient storage proteins in the silkworm hemolymph. Previous studies have shown that 30K proteins are involved in the anti-fungal immunity; however, the molecular mechanism involved in this immunity remains unclear. Methods: We investigated the transcriptional expression of five 30K proteins, including BmLP1, BmLP2, BmLP3, BmLP4, and BmLP7. The five recombinant 30K proteins were expressed in an Escherichia coli expression system, and used for binding assays with fungal cells and hemocytes. Results: The transcriptional expression showed that the five 30K proteins were significantly upregulated after injection of pathogen-associated molecular patterns to the fifth instar larvae, indicating the possibility of their involvement in immune response. The binding assay showed that only BmLP1 and BmLP4 can bind to both fungal cells and silkworm hemocytes. Furthermore, we found that BmLP1-coated and BmLP4-coated agarose beads promote encapsulation of hemocytes in vitro. The hemocyte encapsulation was blocked when the BmLP1-coated beads were preincubated with BmLP1 specific polyclonal antibodies. Conclusions: These results demonstrate that 30K proteins are involved in the cellular immunity of silkworms by acting as pattern recognition molecules to directly recruit hemocytes to the fungal surface.