bluuebunny/crossref_metadata_2025 huggingface dataset:
An Architecture for Biometric Electronic Identification Document System Based on Blockchain †
Authors: Páez Rafael , Pérez Manuel , Ramírez Gustavo , Montes Juan , Bouvarel Lucas
Published: January 2020

This paper proposes an architecture for biometric electronic identification document (e-ID) system based on Blockchain for citizens identity verification in transactions corresponding to the notary, registration, tax declaration and payment, basic health services and registration of economic activities, among others. To validate the user authentication, a biometric e-ID system is used to avoid spoofing and related attacks. Also, to validate the document a digital certificate is used with the corresponding public and private key for each citizen by using a user’s PIN. The proposed transaction validation process was implemented on a Blockchain system in order to record and verify the transactions made by all citizens registered in the electoral census, which guarantees security, integrity, scalability, traceability, and no-ambiguity. Additionally, a Blockchain network architecture is presented in a distributed and decentralized way including all the nodes of the network, database and government entities such as national register and notary offices. The results of the application of a new consensus algorithm to our Blockchain network are also presented showing mining time, memory and CPU usage when the number of transactions scales up.
Effects of Dried Olive Pulp Dietary Supplementation on Quality Characteristics and Antioxidant Capacity of Pig Meat
Authors: Tsala Anna , Mpekelis Vasilios , Karvelis Giannis , Tsikakis Panagiotis , Goliomytis Michael , Simitzis Panagiotis
Published: January 2020

Olive pulp belongs to agro-industrial by-products, and its addition into livestock diets generally result in neutral or positive effects on performance. However, the data concerning the effects of olive by-products on pork meat characteristics are scarce. The aim of this preliminary study was therefore to examine the effects of dried olive pulp (DOP) dietary supplementation on quality parameters and oxidative stability of pig meat. Twenty finishing pigs were allocated to two groups: C that was provided with a control diet, and DOP that was fed with an isonitrogenous and isoenergetic diet supplemented with dried olive pulp at the level of 50 g/kg or 5%. As indicated, cold carcass weight, pH, lightness, redness, cooking loss, and tenderness were not influenced by DOP inclusion. Only meat yellowness (a*) was significantly decreased in DOP meat samples. Moreover, meat oxidation values tended to decrease in the DOP group after one day, but no further differences were observed after four, six, and eight days of refrigerated storage. It can be concluded that DOP dietary supplementation could be recommended as a feasible approach, especially in the Mediterranean region to reduce diet formulation costs, while no significant side effects on pork meat quality are observed.
Powder and Reconstituted Properties of Commercial Infant and Follow-On Formulas
Authors: Murphy Eoin G. , Regost Nicolas E. , Roos Yrjö H. , Fenelon Mark A.
Published: January 2020

The physical properties of 15 commercially available infant formulas (IF) and follow-on (FO) formulas were analysed. Powders made with intact milk proteins were classified into two groups; Type I—homogenous mixtures of milk powder particles (n = 6); and Type II—heterogeneous mixtures of milk powder particles and tomahawk-shaped α-lactose monohydrate crystals (n = 6). Powders made using hydrolysed proteins were classified as Type III powders (n = 3). Type II powders exhibited similar flow characteristics to Type I powders despite having significantly (p < 0.05) smaller particle size, lower circularity, and greater elongation. Type III powders exhibited lowest particles size, highest surface free fat, and poorest flow properties (p < 0.05 for all). Upon reconstitution of powders (12.5% w/w), no significant difference (p < 0.05) in apparent viscosity was observed between Type I and II powders. Reconstituted Type III powders had relatively poor stability to separation compared to Type I and II powders, caused by large starch granules and/or poor emulsification by hydrolysed proteins. Overall, this study illustrated the range of physical behaviour and structures present in commercial IF powders. In particular, the effect of dry addition of lactose and the hydrolysis of protein were found to have major effects on physical properties.
Textural Effects on Perceived Satiation and Ad Libitum Intake of Potato Chips in Males and Females
Authors: Cahayadi Jimmy , Leong Sze Ying , Oey Indrawati , Peng Mei
Published: January 2020

Food texture plays a critical role in influencing an individual’s perceived satiation and ad libitum intake. It remains unclear, however, whether such textural changes can also affect snack consumption. This study aimed to address this question by testing for changes in perceived satiation and ad libitum intake of two types of potato chips with varying hardness. In addition, the observed effect was compared across gender groups. With a crossover design, 74 participants (31 females and 43 males) performed a food consumption task for two types of chips produced from potatoes that were either untreated or treated with pulsed electric fields (PEF) technology. Sensory analyses indicated that these two types of chips had comparable hedonic value, despite a clear textural difference. Across sexes, the results revealed a significant difference in perceived satiation for the two types of chips (p = 0.009), but not in intake. By contrast, analyses of males alone revealed that male participants rated PEF-treated chips to be more satiating than the control chips and correspondingly consumed less (p < 0.05). Overall, findings from the study suggest that modifications of food texture can be a helpful tool in reducing energy intake from snack consumption. The contrasting results from different gender groups highlight the importance of considering gender effects in studies of eating behaviour.
Ovarian Transcriptomic Analyses in the Urban Human Health Pest, the Western Black Widow Spider
Authors: Miles Lindsay S. , Ayoub Nadia A. , Garb Jessica E. , Haney Robert A. , Verrelli Brian C.
Published: January 2020

Due to their abundance and ability to invade diverse environments, many arthropods have become pests of economic and health concern, especially in urban areas. Transcriptomic analyses of arthropod ovaries have provided insight into life history variation and fecundity, yet there are few studies in spiders despite their diversity within arthropods. Here, we generated a de novo ovarian transcriptome from 10 individuals of the western black widow spider (Latrodectus hesperus), a human health pest of high abundance in urban areas, to conduct comparative ovarian transcriptomic analyses. Biological processes enriched for metabolism—specifically purine, and thiamine metabolic pathways linked to oocyte development—were significantly abundant in L. hesperus. Functional and pathway annotations revealed overlap among diverse arachnid ovarian transcriptomes for highly-conserved genes and those linked to fecundity, such as oocyte maturation in vitellogenin and vitelline membrane outer layer proteins, hormones, and hormone receptors required for ovary development, and regulation of fertility-related genes. Comparative studies across arachnids are greatly needed to understand the evolutionary similarities of the spider ovary, and here, the identification of ovarian proteins in L. hesperus provides potential for understanding how increased fecundity is linked to the success of this urban pest.
Review of Watershed-Scale Water Quality and Nonpoint Source Pollution Models
Authors: Yuan Lifeng , Sinshaw Tadesse , Forshay Kenneth J.
Published: January 2020

Watershed-scale nonpoint source (NPS) pollution models have become important tools to understand, evaluate, and predict the negative impacts of NPS pollution on water quality. Today, there are many NPS models available for users. However, different types of models possess different form and structure as well as complexity of computation. It is difficult for users to select an appropriate model for a specific application without a clear understanding of the limitations or strengths for each model or tool. This review evaluates 14 more commonly used watershed-scale NPS pollution models to explain how and when the application of these different models are appropriate for a given effort. The models that are assessed have a wide range of capacities that include simple models used as rapid screening tools (e.g., Long-Term Hydrologic Impact Assessment (L-THIA) and Nonpoint Source Pollution and Erosion Comparison Tool (N-SPECT/OpenNSPECT)), medium-complexity models that require detail data input and limited calibration (e.g., Generalized Watershed Loading Function (GWLF), Loading Simulation Program C (LSPC), Source Loading and Management Model (SLAMM), and Watershed Analysis Risk Management Frame (WARMF)), complex models that provide sophisticated simulation for NPS pollution processes with intensive data and rigorous calibration (e.g., Agricultural Nonpoint Source pollution model (AGNPS/AnnAGNPS), Soil and Water Assessment Tool (SWAT), Stormwater Management Model (SWMM), and Hydrologic Simulation Program Fortran (HSPF)), and modeling systems that integrate various sub-models and tools, and contain the highest complexity to solve all phases of hydrologic, hydraulic, and chemical dynamic processes (e.g., Automated Geospatial Watershed Assessment Tool (AGWA), Better Assessment Science Integrating Point and Nonpoint Sources (BASINS) and Watershed Modeling System (WMS)). This assessment includes model intended use, components or capabilities, suitable land-use type, input parameter type, spatial and temporal scale, simulated pollutants, strengths and limitations, and software availability. Understanding the strengths and weaknesses of each watershed-scale NPS model will lead to better model selection for suitability and help to avoid misinterpretation or misapplication in practice. The article further explains the crucial criteria for model selection, including spatial and temporal considerations, calibration and validation, uncertainty analysis, and future research direction of NPS pollution models. The goal of this work is to provide accurate and concise insight for watershed managers and planners to select the best-suited model to reduce the harm of NPS pollution to watershed ecosystems.
From Braemar to Hollywood: The American Appropriation of Robert Louis Stevenson’s Pirates
Authors: Hill Richard J. , Eidam Laura
Published: January 2020

The pirate tropes that pervade popular culture today can be traced in large part to Robert Louis Stevenson’s 1883 novel, Treasure Island. However, it is the novel’s afterlife on film that has generated fictional pirates as we now understand them. By tracing the transformation of the author’s pirate captain, Long John Silver, from N. C. Wyeth’s illustrations (1911) through the cinematic performances of Wallace Beery (1934) and Robert Newton (1950), this paper demonstrates that the films have created a quintessentially “American pirate”—a figure that has necessarily evolved in response to differences in medium, the performances of the leading actors, and filmgoers’ expectations. Comparing depictions of Silver’s dress, physique, and speech patterns, his role vis-à-vis Jim Hawkins, each adaptation’s narrative point of view, and Silver’s departure at the end of the films reveals that while the Silver of the silver screen may appear to represent a significant departure from the text, he embodies a nuanced reworking of and testament to the author’s original.
Autism Spectrum Disorders: Prenatal Genetic Testing and Abortion Decision-Making among Taiwanese Mothers of Affected Children
Authors: Chen Wei-Ju , Zhao Shixi , Huang Tse-Yang , Kwok Oi-Man , Chen Lei-Shih
Published: January 2020

With the rapid growing rate of autism spectrum disorders (ASDs), prenatal genetic testing (PGT) has been offered to detect various genomic disorders, including ASD, in Taiwan. However, disparities exist in this area, as there is limited research on factors associated with PGT utilization and relevant decision-making that may guide the regulations and ethical guidelines for culturally appropriate PGT services in Taiwan. This study proposed a comprehensively integrated theoretical framework for examining the intention to undergo PGT to detect ASD susceptibility genes and subsequent abortion decision-making among Taiwanese mothers of children affected by ASD. Survey data from 333 mothers of children with ASD in 236 elementary schools with special education services in Taiwan were collected and analyzed using structural equation modeling. Approximately two-thirds of the participants (66.6%) would undergo PGT to detect ASD susceptibility genes; more than half (53.1%) would terminate the hypothetically ASD-affected pregnancy. Abortion intention was associated with age, religion, attitudes toward PGT for detecting ASD susceptibility genes, and willingness to undergo such PGT. This study explores the potential impacts of PGT on Taiwanese society, and the findings are applicable to countries heavily influenced by Chinese culture, areas with Asian immigrants, and Western countries with such PGT services and/or research available.
Forest Roles in Particle Removal during Spring Dust Storms on Transport Path
Authors: Wei Wenjun , Wang Bing , Niu Xiang
Published: January 2020

The transpacific transport of mineral dust often occurs in spring, and a large amount of aeolian dust is deposited in the Asian continent and north Pacific. Moreover, a heterogeneous reaction occurs when dust particles are mixed with man-made pollution gases and particles. In the present study, atmospheric PM10 and PM2.5 concentrations were investigated, and a scanning electron microscope and an X-ray energy spectrometer were used to analyze the effects of dust resistance and capture by forests. It showed that (1) the PM2.5 and PM10 concentrations during a dust storm, on sunny days, and during light pollution periods, were higher in the non-forest covered area (NFC area) than in the forest covered area (FC area), except during heavy pollution events; which suggests that the forests have a strong effect on dust resistance; (2) the PM reduction efficiency of forests was highest on sunny days, followed by light pollution periods, heavy pollution periods, and during the dust storm; (3) after the dust storm, TSP captured by leaves significantly increased, especially for the broadleaved tree species; and the particulates number in the grooves on leaves’ surface increased particularly sharply. This study will help improve the dust resistance and retention efficiency of forest shelterbelt projects during dust storms.
Rome, a Policy without Politics: The Participatory Process for a Metropolitan Scale Food Policy
Authors: Mazzocchi Giampiero , Marino Davide
Published: January 2020

In light of the challenges that all cities face today, food is offered as a prism through which to read and intervene on various areas that affect the quality of life of the population: circular economy, urban metabolism, social relations, economies, and food quality. In the Roman context, in recent years, numerous initiatives have revitalized the debate on food and brought the discussion to the center of the interest of an ever-increasing number of citizens. However, these experiences appear unrelated and there is a lack of coordination and political coherence. Faced with this evidence, starting from a territorial analysis, this contribution analyzes the process that led a local group of stakeholders to formulate a proposal for a food policy for the city of Rome. The proposal contains a series of possible actions that aim, on the one hand, to recompose the relations between the city and its territory, with a view to re-localization and re-territorialization of agro-food productions and, on the other hand, to reconnect the economic and social relations that the industrialization of food chains has compromised. The network analysis of the bottom-up process, which mainly investigates networking and negotiation skills between various interests, is carried out and related to a careful analysis of the food system in the Roman context. Furthermore, an overview of the state of the art of urban food policies in Italy has been provided to better contextualize the study case. The findings show actors and topics involved in the process, identifying further development towards a more comprehensive participatory process for a systemic food strategy at the metropolitan level.
Effect of Chemotherapy, Laparoscopy, and Cytology on Stage IC Ovarian Clear Cell Carcinoma: A Long-Term, Single-Center Study
Authors: Chang Hao-Ting , Chiu Mei-Ling , Wang Tao-Yuean , Chen Tzu-Chien , Chang Chih-Long , Su Tsung-Hsien , Wang Kuo-Gong , Wang Kung-Liahng , Yang Yuh-Cheng , Chen Jen-Ruei
Published: January 2020

Ovarian clear cell carcinoma (OCCC) is the second common histology of epithelial ovarian cancer in Taiwan. Stage IC is common, especially during minimally invasive surgery. Adjuvant chemotherapy in stage IC OCCC is unavoidable, and paclitaxel-based chemotherapy in Taiwan is self-paid. However, surgical spillage from minimally invasive surgery as a cause of unfavorable prognosis is still uncertain. The information of patients with stage IC OCCC, corresponding to a period of January 1995 to December 2016, was retrospectively collected following a chart and pathology review. Data regarding surgical methods, cytology status, regimens of adjuvant chemotherapy, survivorship, progression-free survival (PFS), and overall survival (OS) period were analyzed. In total, 88 patients were analyzed, and 64 and 24 patients were treated with paclitaxel- and nonpaclitaxel-based chemotherapy, respectively. Recurrence was identical between the two groups: PFS (47.5 ± 41.36 versus 54.0 ± 53.9 months, p = 0.157) and OS (53.5 ± 38.14 versus 79.0 ± 49.42 months, p = 0.070). Of the 88 patients, 12 had undergone laparoscopy for histological confirmation before complete open staging surgery; however, their PFS (49.5 ± 46.84 versus 49.0 ± 35.55 months, p = 0.719) and OS (56.5 ± 43.4 versus 51.0 ± 32.77 months, p = 0.600) were still comparable. Cytology results were only available for 51 patients, and positive washing cytology results seemed to worsen PFS (p = 0.026) but not OS (p = 0.446). In conclusion, adjuvant nonpaclitaxel chemotherapy and laparoscopic tumor spillage before the staging operation did not worsen the outcome in stage IC OCCC. Positive washing cytology has a negative effect on PFS but not on OS.
Circular RNA-Centered Regulatory Networks in the Physiopathology of Cardiovascular Diseases
Authors: Gabriel André F. , Costa Marina C. , Enguita Francisco J.
Published: January 2020

Non-coding regulatory RNAs are generated as a core output of the eukaryotic genomes, being essential players in cell biology. At the organism level, they are key functional actors in those tissues and organs with limited proliferation capabilities such as the heart. The role of regulatory networks mediated by non-coding RNAs in the pathophysiology of cardiovascular conditions is starting to be unveiled. However, a deeper knowledge of the functional interactions among the diverse non-coding RNA families and their phenotypic consequences is required. This review presents the current knowledge about the functional crosstalk between circRNAs and other biomolecules in the framework of the cardiovascular diseases.
Supporting Cell-Based Tendon Therapy: Effect of PDGF-BB and Ascorbic Acid on Rabbit Achilles Tenocytes In Vitro
Authors: Evrova Olivera , Kellenberger Damian , Calcagni Maurizio , Vogel Viola , Buschmann Johanna
Published: January 2020

Cell-based tendon therapies with tenocytes as a cell source need effective tenocyte in vitro expansion before application for tendinopathies and tendon injuries. Supplementation of tenocyte culture with biomolecules that can boost proliferation and matrix synthesis is one viable option for supporting cell expansion. In this in vitro study, the impacts of ascorbic acid or PDGF-BB supplementation on rabbit Achilles tenocyte culture were studied. Namely, cell proliferation, changes in gene expression of several ECM and tendon markers (collagen I, collagen III, fibronectin, aggrecan, biglycan, decorin, ki67, tenascin-C, tenomodulin, Mohawk, α-SMA, MMP-2, MMP-9, TIMP1, and TIMP2) and ECM deposition (collagen I and fibronectin) were assessed. Ascorbic acid and PDGF-BB enhanced tenocyte proliferation, while ascorbic acid significantly accelerated the deposition of collagen I. Both biomolecules led to different changes in the gene expression profile of the cultured tenocytes, where upregulation of collagen I, Mohawk, decorin, MMP-2, and TIMP-2 was observed with ascorbic acid, while these markers were downregulated by PDGF-BB supplementation. Vice versa, there was an upregulation of fibronectin, biglycan and tenascin-C by PDGF-BB supplementation, while ascorbic acid led to a downregulation of these markers. However, both biomolecules are promising candidates for improving and accelerating the in vitro expansion of tenocytes, which is vital for various tendon tissue engineering approaches or cell-based tendon therapy.
Unveiling the Molecular Basis of the Noonan Syndrome-Causing Mutation T42A of SHP2
Authors: Toto Angelo , Malagrinò Francesca , Visconti Lorenzo , Troilo Francesca , Gianni Stefano
Published: January 2020

Noonan syndrome (NS) is a genetic disorder caused by the hyperactivation of the RAS-MAPK molecular pathway. About 50% of NS cases are caused by mutations affecting the SHP2 protein, a multi-domain phosphatase with a fundamental role in the regulation of the RAS-MAPK pathway. Most NS-causing mutations influence the stability of the inactive form of SHP2. However, one NS-causing mutation, namely T42A, occurs in the binding pocket of the N-SH2 domain of the protein. Here, we present a quantitative characterization of the effect of the T42A mutation on the binding of the N-terminal SH2 domain of SHP2 with a peptide mimicking Gab2, a fundamental interaction that triggers the activation of the phosphatase in the cellular environment. Our results show that whilst the T42A mutation does not affect the association rate constant with the ligand, it causes a dramatic increase of the affinity for Gab2. This effect is due to a remarkable decrease of the microscopic dissociation rate constant of over two orders of magnitudes. In an effort to investigate the molecular basis of the T42A mutation in causing Noonan syndrome, we also compare the experimental results with a more conservative variant, T42S. Our findings are discussed in the context of the structural data available on SHP2.
Antagonistic Autoantibodies to Insulin-Like Growth Factor-1 Receptor Associate with Poor Physical Strength
Authors: Schwiebert Christian , Kühnen Peter , Becker Niels-Peter , Welsink Tim , Keller Theresa , Minich Waldemar B. , Wiegand Susanna , Schomburg Lutz
Published: January 2020

Natural autoantibodies to the IGF1 receptor (IGF1R-aAb) have been described in relation to Graves’ ophthalmopathy. Other physiological roles of natural IGF1R-aAb are not known. We hypothesized that IGF1R-aAb may be related to muscle development. Serum samples (n = 408) from young overweight subjects (n = 143) were collected during a lifestyle intervention study. Anthropometric parameters, along with leptin, IGF1 and IGF1R-aAb concentrations, were analyzed, and the subjects were categorized into positive or negative for IGF1R-aAb. Eleven out of 143 subjects (7.7%) were positive for IGF1R-aAb. Identified IGF1R-aAb were molecularly characterized and showed antagonistic activity in vitro impairing IGF1-mediated IGF1R activation. Mean body weight, height or age were similar between IGF1R-aAb-positive and -negative subjects, but IGF1 concentrations differed. Jumping ability, as well as right and left handgrip strengths, were lower in the IGF1R-aAb-positive as compared to the IGF1R-aAb-negative subjects. We conclude that natural IGF1R-aAb are detectable in apparently healthy subjects and are capable of antagonizing IGF1-dependent IGF1R activation. Moreover, the presence of IGF1R-aAb is associated with poor physical strength. Although the causality of this association is unclear, the data imply a potential influence of IGF1R autoimmunity on muscle development.
Identification of 56 Proteins Involved in Embryo–Maternal Interactions in the Bovine Oviduct
Authors: Banliat Charles , Tsikis Guillaume , Labas Valérie , Teixeira-Gomes Ana-Paula , Com Emmanuelle , Lavigne Régis , Pineau Charles , Guyonnet Benoit , Mermillod Pascal , Saint-Dizier Marie
Published: January 2020

The bovine embryo develops in contact with the oviductal fluid (OF) during the first 4–5 days of pregnancy. The aim of this study was to decipher the protein interactions occurring between the developing embryo and surrounding OF. In-vitro produced 4–6 cell and morula embryos were incubated or not (controls) in post-ovulatory OF (OF-treated embryos) and proteins were then analyzed and quantified by high resolution mass spectrometry (MS) in both embryo groups and in OF. A comparative analysis of MS data allowed the identification and quantification of 56 embryo-interacting proteins originated from the OF, including oviductin (OVGP1) and several annexins (ANXA1, ANXA2, ANXA4) as the most abundant ones. Some embryo-interacting proteins were developmental stage-specific, showing a modulating role of the embryo in protein interactions. Three interacting proteins (OVGP1, ANXA1 and PYGL) were immunolocalized in the perivitelline space and in blastomeres, showing that OF proteins were able to cross the zona pellucida and be taken up by the embryo. Interacting proteins were involved in a wide range of functions, among which metabolism and cellular processes were predominant. This study identified for the first time a high number of oviductal embryo-interacting proteins, paving the way for further targeted studies of proteins potentially involved in the establishment of pregnancy in cattle.
Maternal Obesity Alters Placental Cell Cycle Regulators in the First Trimester of Human Pregnancy: New Insights for BRCA1
Authors: Hoch Denise , Bachbauer Martina , Pöchlauer Caroline , Algaba-Chueca Francisco , Tandl Veronika , Novakovic Boris , Megia Ana , Gauster Martin , Saffery Richard , Glasner Andreas , Desoye Gernot , Majali-Martinez Alejandro
Published: January 2020

In the first trimester of pregnancy, placental development involves a wide range of cellular processes. These include trophoblast proliferation, fusion, and differentiation, which are dependent on tight cell cycle control. The intrauterine environment affects placental development, which also includes the trophoblast cell cycle. In this work, we focus on maternal obesity to assess whether an altered intrauterine milieu modulates expression and protein levels of placental cell cycle regulators in early human pregnancy. For this purpose, we use first trimester placental tissue from lean and obese women (gestational week 5+0–11+6, n = 58). Using a PCR panel, a cell cycle protein array, and STRING database analysis, we identify a network of cell cycle regulators increased by maternal obesity in which breast cancer 1 (BRCA1) is a central player. Immunostaining localizes BRCA1 predominantly to the villous and the extravillous cytotrophoblast. Obesity-driven BRCA1 upregulation is not able to be explained by DNA methylation (EPIC array) or by short-term treatment of chorionic villous explants at 2.5% oxygen with tumor necrosis factor α (TNF-α) (50 mg/mL), leptin (100 mg/mL), interleukin 6 (IL-6) (100 mg/mL), or high glucose (25 nM). Oxygen tension rises during the first trimester, but this change in vitro has no effect on BRCA1 (2.5% and 6.5% O2). We conclude that maternal obesity affects placental cell cycle regulation and speculate this may alter placental development.
Characterization of Human Induced Pluripotent Stem Cell-Derived Hepatocytes with Mature Features and Potential for Modeling Metabolic Diseases
Authors: Holmgren Gustav , Ulfenborg Benjamin , Asplund Annika , Toet Karin , Andersson Christian X , Hammarstedt Ann , Hanemaaijer Roeland , Küppers-Munther Barbara , Synnergren Jane
Published: January 2020

There is a strong anticipated future for human induced pluripotent stem cell-derived hepatocytes (hiPS-HEP), but so far, their use has been limited due to insufficient functionality. We investigated the potential of hiPS-HEP as an in vitro model for metabolic diseases by combining transcriptomics with multiple functional assays. The transcriptomics analysis revealed that 86% of the genes were expressed at similar levels in hiPS-HEP as in human primary hepatocytes (hphep). Adult characteristics of the hiPS-HEP were confirmed by the presence of important hepatocyte features, e.g., Albumin secretion and expression of major drug metabolizing genes. Normal energy metabolism is crucial for modeling metabolic diseases, and both transcriptomics data and functional assays showed that hiPS-HEP were similar to hphep regarding uptake of glucose, low-density lipoproteins (LDL), and fatty acids. Importantly, the inflammatory state of the hiPS-HEP was low under standard conditions, but in response to lipid accumulation and ER stress the inflammation marker tumor necrosis factor α (TNFα) was upregulated. Furthermore, hiPS-HEP could be co-cultured with primary hepatic stellate cells both in 2D and in 3D spheroids, paving the way for using these co-cultures for modeling non-alcoholic steatohepatitis (NASH). Taken together, hiPS-HEP have the potential to serve as an in vitro model for metabolic diseases. Furthermore, differently expressed genes identified in this study can serve as targets for future improvements of the hiPS-HEP.
The Influence of Met Receptor Level on HGF-Induced Glycolytic Reprogramming in Head and Neck Squamous Cell Carcinoma
Authors: Boschert Verena , Klenk Nicola , Abt Alexander , Janaki Raman Sudha , Fischer Markus , Brands Roman C. , Seher Axel , Linz Christian , Müller-Richter Urs D. A. , Bischler Thorsten , Hartmann Stefan
Published: January 2020

Head and neck squamous cell carcinoma (HNSCC) is known to overexpress a variety of receptor tyrosine kinases, such as the HGF receptor Met. Like other malignancies, HNSCC involves a mutual interaction between the tumor cells and surrounding tissues and cells. We hypothesized that activation of HGF/Met signaling in HNSCC influences glucose metabolism and therefore substantially changes the tumor microenvironment. To determine the effect of HGF, we submitted three established HNSCC cell lines to mRNA sequencing. Dynamic changes in glucose metabolism were measured in real time by an extracellular flux analyzer. As expected, the cell lines exhibited different levels of Met and responded differently to HGF stimulation. As confirmed by mRNA sequencing, the level of Met expression was associated with the number of upregulated HGF-dependent genes. Overall, Met stimulation by HGF leads to increased glycolysis, presumably mediated by higher expression of three key enzymes of glycolysis. These effects appear to be stronger in Methigh-expressing HNSCC cells. Collectively, our data support the hypothesized role of HGF/Met signaling in metabolic reprogramming of HNSCC.
Novel Segmented Concentration Addition Method to Predict Mixture Hormesis of Chlortetracycline Hydrochloride and Oxytetracycline Hydrochloride to Aliivibrio fischeri
Authors: Ge Huilin , Zhou Min , Lv Daizhu , Wang Mingyue , Xie Defang , Yang Xinfeng , Dong Cunzhu , Li Shuhuai , Lin Peng
Published: January 2020

Hormesis is a concentration-response phenomenon characterized by low-concentration stimulation and high-concentration inhibition, which typically has a nonmonotonic J-shaped concentration-response curve (J-CRC). The concentration addition (CA) model is the gold standard for studying mixture toxicity. However, the CA model had the predictive blind zone (PBZ) for mixture J-CRC. To solve the PBZ problem, we proposed a segmented concentration addition (SCA) method to predict mixture J-CRC, which was achieved through fitting the left and right segments of component J-CRC and performing CA prediction subsequently. We selected two model compounds including chlortetracycline hydrochloride (CTCC) and oxytetracycline hydrochloride (OTCC), both of which presented J-CRC to Aliivibrio fischeri (AVF). The seven binary mixtures (M1–M7) of CTCC and OTCC were designed according to their molar ratios of 12:1, 10:3, 8:5, 1:1, 5:8, 3:10, and 1:12 referring to the direct equipartition ray design. These seven mixtures all presented J-CRC to AVF. Based on the SCA method, we obtained mixture maximum stimulatory effect concentration (ECm) and maximum stimulatory effect (Em) predicted by SCA, both of which were not available for the CA model. The toxicity interactions of these mixtures were systematically evaluated by using a comprehensive approach, including the co-toxicity coefficient integrated with confidence interval method (CTCICI), CRC, and isobole analysis. The results showed that the interaction types were additive and antagonistic action, without synergistic action. In addition, we proposed the cross point (CP) hypothesis for toxic interactive mixtures presenting J-CRC, that there was generally a CP between mixture observed J-CRC and CA predicted J-CRC; the relative positions of observed and predicted CRCs on either side of the CP would exchange, but the toxic interaction type of mixtures remained unchanged. The CP hypothesis needs to be verified by more mixtures, especially those with synergism. In conclusion, the SCA method is expected to have important theoretical and practical significance for mixture hormesis.
Calmodulin-Mediated Regulation of Gap Junction Channels
Authors: Peracchia Camillo
Published: January 2020

Evidence that neighboring cells uncouple from each other as one dies surfaced in the late 19th century, but it took almost a century for scientists to start understanding the uncoupling mechanism (chemical gating). The role of cytosolic free calcium (Ca2+i) in cell–cell channel gating was first reported in the mid-sixties. In these studies, only micromolar [Ca2+]i were believed to affect gating—concentrations reachable only in cell death, which would discard Ca2+i as a fine modulator of cell coupling. More recently, however, numerous researchers, including us, have reported the effectiveness of nanomolar [Ca2+]i. Since connexins do not have high-affinity calcium sites, the effectiveness of nanomolar [Ca2+]i suggests the role of Ca-modulated proteins, with calmodulin (CaM) being most obvious. Indeed, in 1981 we first reported that a CaM-inhibitor prevents chemical gating. Since then, the CaM role in gating has been confirmed by studies that tested it with a variety of approaches such as treatments with CaM-inhibitors, inhibition of CaM expression, expression of CaM mutants, immunofluorescent co-localization of CaM and gap junctions, and binding of CaM to peptides mimicking connexin domains identified as CaM targets. Our gating model envisions Ca2+-CaM to directly gate the channels by acting as a plug (“Cork” gating model), and probably also by affecting connexin conformation.
Pharmacological Chaperones: A Therapeutic Approach for Diseases Caused by Destabilizing Missense Mutations
Authors: Liguori Ludovica , Monticelli Maria , Allocca Mariateresa , Hay Mele Bruno , Lukas Jan , Cubellis Maria Vittoria , Andreotti Giuseppina
Published: January 2020

The term “pharmacological chaperone” was introduced 20 years ago. Since then the approach with this type of drug has been proposed for several diseases, lysosomal storage disorders representing the most popular targets. The hallmark of a pharmacological chaperone is its ability to bind a protein specifically and stabilize it. This property can be beneficial for curing diseases that are associated with protein mutants that are intrinsically active but unstable. The total activity of the affected proteins in the cell is lower than normal because they are cleared by the quality control system. Although most pharmacological chaperones are reversible competitive inhibitors or antagonists of their target proteins, the inhibitory activity is neither required nor desirable. This issue is well documented by specific examples among which those concerning Fabry disease. Direct specific binding is not the only mechanism by which small molecules can rescue mutant proteins in the cell. These drugs and the properly defined pharmacological chaperones can work together with different and possibly synergistic modes of action to revert a disease phenotype caused by an unstable protein.
The Mitochondrial Ca2+ Overload via Voltage-Gated Ca2+ Entry Contributes to an Anti-Melanoma Effect of Diallyl Trisulfide
Authors: Nakagawa Chinatsu , Suzuki-Karasaki Manami , Suzuki-Karasaki Miki , Ochiai Toyoko , Suzuki-Karasaki Yoshihiro
Published: January 2020

Allium vegetables such as garlic (Allium sativum L.) are rich in organosulfur compounds that prevent human chronic diseases, including cancer. Of these, diallyl trisulfide (DATS) exhibits anticancer effects against a variety of tumors, including malignant melanoma. Although previous studies have shown that DATS increases intracellular calcium (Ca2+) in different cancer cell types, the role of Ca2+ in the anticancer effect is obscure. In the present study, we investigated the Ca2+ pathways involved in the anti-melanoma effect. We used melittin, the bee venom that can activate a store-operated Ca2+ entry (SOCE) and apoptosis, as a reference. DATS increased apoptosis in human melanoma cell lines in a Ca2+-dependent manner. It also induced mitochondrial Ca2+ (Ca2+mit) overload through intracellular and extracellular Ca2+ fluxes independently of SOCE. Strikingly, acidification augmented Ca2+mit overload, and Ca2+ channel blockers reduced the effect more significantly under acidic pH conditions. On the contrary, acidification mitigated SOCE and Ca2+mit overload caused by melittin. Finally, Ca2+ channel blockers entirely inhibited the anti-melanoma effect of DATS. Our findings suggest that DATS explicitly evokes Ca2+mit overload via a non-SOCE, thereby displaying the anti-melanoma effect.
Expression, Purification, and Characterization of Anti-Zika virus Envelope Protein: Polyclonal and Chicken-Derived Single Chain Variable Fragment Antibodies
Authors: Mwale Pharaoh Fellow , Lee Chi-Hsin , Lin Liang-Tzung , Leu Sy-Jye , Huang Yun-Ju , Chiang Liao-Chun , Mao Yan-Chiao , Yang Yi-Yuan
Published: January 2020

Zika virus (ZIKV) is a new and emerging virus that has caused outbreaks worldwide. The virus has been linked to congenital neurological malformations in neonates and Guillain–Barré syndrome in adults. Currently there are no effective vaccines available. As a result, there is a great need for ZIKV treatment. In this study, we developed single chain variable fragment (scFv) antibodies that target the ZIKV envelope protein using phage display technology. We first induced an immune response in white leghorn laying hens against the ZIKV envelope (E) protein. Chickens were immunized and polyclonal immunoglobulin yolk (IgY) antibodies were extracted from egg yolks. A high-level titer of anti-ZIKV_E IgY antibodies was detected using enzyme-linked immunosorbent assay (ELISA) after the third immunization. The titer persisted for at least 9 weeks. We constructed two antibody libraries that contained 5.3 × 106 and 4.5 × 106 transformants. After biopanning, an ELISA phage assay confirmed the enrichment of specific clones. We randomly selected 26 clones that expressed ZIKV scFv antibodies and classified them into two groups, short-linker and long-linker. Of these, four showed specific binding activities toward ZIKV_E proteins. These data suggest that the polyclonal and monoclonal scFv antibodies have the diagnostic or therapeutic potential for ZIKV.
Viability of Neural Networks for Core Technologies for Resource-Scarce Languages
Authors: Loubser Melinda , Puttkammer Martin J.
Published: January 2020

In this paper, the viability of neural network implementations of core technologies (the focus of this paper is on text technologies) for 10 resource-scarce South African languages is evaluated. Neural networks are increasingly being used in place of other machine learning methods for many natural language processing tasks with good results. However, in the South African context, where most languages are resource-scarce, very little research has been done on neural network implementations of core language technologies. In this paper, we address this gap by evaluating neural network implementations of four core technologies for ten South African languages. The technologies we address are part of speech tagging, named entity recognition, compound analysis and lemmatization. Neural architectures that performed well on similar tasks in other settings were implemented for each task and the performance was assessed in comparison with currently used machine learning implementations of each technology. The neural network models evaluated perform better than the baselines for compound analysis, are viable and comparable to the baseline on most languages for POS tagging and NER, and are viable, but not on par with the baseline, for Afrikaans lemmatization.
Cost Assessment of Five Different Maize Grain Handling Techniques to Reduce Postharvest Losses from Insect Contamination
Authors: Darfour Bernard , Rosentrater Kurt A.
Published: January 2020

Farmers in developing nations encounter high postharvest losses mainly attributable to the lack of modern techniques for threshing, cleaning, grading, and grain storage. Mechanized handling of grain in developing countries is rare, although the technology is effective against insects and pest infestations. The objective was to evaluate the cost-effectiveness of five grain handling techniques that have the ability to reduce postharvest losses from insect infestation. The five methods were metal silo plus all accessories (m. silo + acc.), metal silo only (m. silo), woven polypropylene plus phosphine (w. PP. + Phos.), woven polypropylene only (w. PP.), and Purdue Improved Crop Storage bags only (PICS). The functional unit used was handling 1 kg of maize grain. The cost analysis of each technique was calculated based on equations using a spreadsheet. The annual capital and operational costs of handling using m. silo + acc. or m. silo were very high, unlike the PICS, w. PP. + Phos., or w. PP. The annual capital and operational costs decreased as production scale increased. Food security (due to reduced insects and pest infestations) and financial prospects of farmers can improve when the grain is mechanically handled with m. silo + acc. or m. silo.
Molecular Phylogeny and Infraordinal Classification of Zoraptera (Insecta)
Authors: Kočárek Petr , Horká Ivona , Kundrata Robin
Published: January 2020

Zoraptera is a small and predominantly tropical insect order with an unresolved higher classification due to the extremely uniform external body morphology. We, therefore, conducted a multigene molecular phylogeny of extant Zoraptera and critically re-evaluated their morphological characters in order to propose a natural infraordinal classification. We recovered a highly-resolved phylogeny with two main clades representing major evolutionary lineages in Zoraptera, for which we propose family ranks. The two families exhibit striking differences in male genitalia and reproductive strategies. Each family contains two subclades (subfamilies) supported by several morphological synapomorphies including the relative lengths of the basal antennomeres, the number and position of metatibial spurs, and the structure of male genitalia. The newly proposed higher classification of Zoraptera includes the family Zorotypidae stat. revid. with Zorotypinae Silvestri, 1913 (Zorotypus stat. revid., Usazoros Kukalova-Peck and Peck, 1993 stat. restit.) and Spermozorinae subfam. nov. (Spermozoros gen. nov.), and Spriralizoridae fam. nov. with Spiralizorinae subfam. nov. (Spiralizoros gen. nov., Scapulizoros gen. nov., Cordezoros gen. nov., Centrozoros Kukalova-Peck and Peck, 1993, stat. restit., Brazilozoros Kukalova-Peck and Peck, 1993, stat. restit.), and Latinozorinae subfam. nov. (Latinozoros Kukalova-Peck and Peck, 1993, stat. restit.). An identification key and morphological diagnoses for all supraspecific taxa are provided.
Circulating Biomarkers of Cell Adhesion Predict Clinical Outcome in Patients with Chronic Heart Failure
Authors: Bouwens Elke , van den Berg Victor J. , Akkerhuis K. Martijn , Baart Sara J. , Caliskan Kadir , Brugts Jasper J. , Mouthaan Henk , van Ramshorst Jan , Germans Tjeerd , Umans Victor A. W. M. , Boersma Eric , Kardys Isabella
Published: January 2020

Cardiovascular inflammation and vascular endothelial dysfunction are involved in chronic heart failure (CHF), and cellular adhesion molecules are considered to play a key role in these mechanisms. We evaluated temporal patterns of 12 blood biomarkers of cell adhesion in patients with CHF. In 263 ambulant patients, serial, tri-monthly blood samples were collected during a median follow-up of 2.2 (1.4–2.5) years. The primary endpoint (PE) was a composite of cardiovascular mortality, HF hospitalization, heart transplantation and implantation of a left ventricular assist device and was reached in 70 patients. We selected the baseline blood samples in all patients, the two samples closest to a PE, or, for event-free patients, the last sample available. In these 567 samples, associations between biomarkers and PE were investigated by joint modelling. The median age was 68 (59–76) years, with 72% men and 74% New York Heart Association class I–II. Repeatedly measured levels of Complement component C1q receptor (C1qR), Cadherin 5 (CDH5), Chitinase-3-like protein 1 (CHI3L1), Ephrin type-B receptor 4 (EPHB4), Intercellular adhesion molecule-2 (ICAM-2) and Junctional adhesion molecule A (JAM-A) were independently associated with the PE. Their rates of change also predicted clinical outcome. Level of CHI3L1 was numerically the strongest predictor with a hazard ratio (HR) (95% confidence interval) of 2.27 (1.66–3.16) per SD difference in level, followed by JAM-A (2.10, 1.42–3.23) and C1qR (1.90, 1.36–2.72), adjusted for clinical characteristics. In conclusion, temporal patterns of C1qR, CDH5, CHI3L1, EPHB4, ICAM2 and JAM-A are strongly and independently associated with clinical outcome in CHF patients.