bluuebunny/crossref_metadata_2025 huggingface dataset:
The Effect of Streaming Potential and Viscous Dissipation in the Heat Transfer Characteristics of Power-Law Nanofluid Flow in a Rectangular Microchannel
Authors: Deng Shuyan , An Quan , Li Mingying
Published: April 2020

The non-Newtonian nanofluid flow becomes increasingly important in enhancing the thermal management efficiency of microscale devices and in promoting the exploration of the thermal-electric energy conversion process. The effect of streaming potential and viscous dissipation in the heat transfer characteristics of power-law nanofluid electrokinetic flow in a rectangular microchannel has been investigated to assist in the development of an energy harvesting system. The electroviscous effect caused by the streaming potential influences the hydrodynamical and thermal characteristics of flow. With the change in constitutive behavior of power-law nanofluid, the viscous dissipation effect is considered. The Poisson–Boltzmann equation, the modified Cauchy momentum equation, and the energy equation were solved. The temperature and heat transfer rate were analytically expressed for Newtonian nanofluid and numerically obtained for power-law nanofluid. The interactive influence of streaming potential, viscous dissipation, and hydrodynamical features of power-law nanofluid on the heat transfer characteristics were studied. The presence of streaming potential tends to reduce the dimensionless bulk mean temperature. The introduction of nanoparticles augments dimensionless temperature difference between channel wall and bulk flow, which decreases the heat transfer rate. The shear thinning nanofluid is more sensitive to the above effects. The temperature is a weak function of the flow behavior index.
mRNA as a Tool for Gene Transfection in 3D Cell Culture for Future Regenerative Therapy
Authors: Uchida Satoshi , Yanagihara Kayoko , Matsui Akitsugu , Kataoka Kazunori , Itaka Keiji
Published: April 2020

A combination of three-dimensional (3D) cell culturing and non-viral gene transfection is promising in improving outcomes of cell transplantation therapy. Herein, gene transfection profiles in 3D cell culture were compared between plasmid DNA (pDNA) and messenger RNA (mRNA) introduction, using mesenchymal stem cell (MSC) 3D spheroids. Green fluorescence protein (GFP) mRNA induced GFP protein expression in 77% of the cells in the spheroids, whereas only 34% of the cells became GFP positive following pDNA introduction. In mechanistic analyses, most of the cells in MSC spheroids were non-dividing, and pDNA failed to induce GFP expression in most of the non-dividing cells. In contrast, both dividing and non-dividing cells became GFP-positive after mRNA introduction, which led to a high overall percentage of GFP-positive cells in the spheroids. Consequently, mRNA encoding an osteogenic factor, runt-related transcription factor 2 (Runx2), allowed in vitro osteogenic differentiation of MSCs in spheroids more efficiently compared to Runx2 pDNA. Conclusively, mRNA exhibits high potential in gene transfection in 3D cell culture, in which the cell division rate is lower than that in monolayer culture, and the combination of mRNA introduction and 3D cell culture is a promising approach to improve outcomes of cell transplantation in future regenerative therapy.
Experimental Characterization and Simulation of Thermoplastic Polymer Flow Hesitation in Thin-Wall Injection Molding Using Direct In-Mold Visualization Technique
Authors: Regi Francesco , Guerrier Patrick , Zhang Yang , Tosello Guido
Published: April 2020

A special mold provided with a glass window was used in order to directly evaluate the flow progression during the filling phase of the injection molding process in a thin-wall cavity and to validate the simulation of the process with particular focus on the hesitation effect. The flow of the polymer was recorded at 500 frames per second using a high-speed camera (HSC). Two unfilled thermoplastic polymers, acrylonitrile butadiene styrene (ABS), and polypropylene (PP), were used to fill two different 50 mm × 18 mm staircase geometry cavities, which were specifically designed to evaluate the hesitation effect with thicknesses of 1500, 1250, 1000, 750, 500 µm (cavity insert no. 1) and 1500, 1200, 900, 600, 300 µm (cavity insert no. 2). In addition to the video recordings, the simulations were validated using the timings and the data obtained by three pressure sensors and two thermocouples located in the cavity. For each injection cycle recorded on camera the machine data were collected to carefully implement the correct boundary conditions in the simulations. The analysis of the video recordings highlighted that flow progression and hesitation were mainly influenced not only by the thickness, but also by the velocity and the material type. The simulation results were in relatively good agreement with the experiments in terms of flow pattern and progression. Filling times were predicted with an average relative error deviation of 2.5% throughout all the section thicknesses of the cavity. Lower accuracies in terms of both filling times and injection pressure were observed at increasingly thinner sections.
A Fluorescent Sensor-Assisted Paper-Based Competitive Lateral Flow Immunoassay for the Rapid and Sensitive Detection of Ampicillin in Hospital Wastewater
Authors: Lin Honggui , Fang Feixiang , Zang Jiahui , Su Jianlong , Tian Qingyuan , Kumar Kankala Ranjith , Lin Xuexia
Published: April 2020

In this study, a convenient assay method has been developed based on labeled functional nucleic acids (H-DNA) and a competitive fluorescent lateral flow immunoassay (CF-LFI) for ampicillin (AMP) detection. Herein, we designed the tunable AMP probes for AMP detection based on the AMP aptamer, and the secondary DNA fragment. The probes can generate tunable signals on the test line (T line) and control line (C line) according to the concentration of AMP. The accuracy of detection was improved by optimizing the tunable AMP probes. Under the optimal conditions, the linear concentration of AMP detection is ranged from 10 to 200 ng/L with a limit of quantitation (LOQ) value of 2.71 ng/L, and the recovery is higher than 80.5 %. Moreover, the developed method shows the potential application for AMP detection in the hospital wastewater.
3D Printed MEMS Technology—Recent Developments and Applications
Authors: Blachowicz Tomasz , Ehrmann Andrea
Published: April 2020

Microelectromechanical systems (MEMS) are of high interest for recent electronic applications. Their applications range from medicine to measurement technology, from microfluidics to the Internet of Things (IoT). In many cases, MEMS elements serve as sensors or actuators, e.g., in recent mobile phones, but also in future autonomously driving cars. Most MEMS elements are based on silicon, which is not deformed plastically under a load, as opposed to metals. While highly sophisticated solutions were already found for diverse MEMS sensors, actuators, and other elements, MEMS fabrication is less standardized than pure microelectronics, which sometimes blocks new ideas. One of the possibilities to overcome this problem may be the 3D printing approach. While most 3D printing technologies do not offer sufficient resolution for MEMS production, and many of the common 3D printing materials cannot be used for this application, there are still niches in which the 3D printing of MEMS enables producing new structures and thus creating elements for new applications, or the faster and less expensive production of common systems. Here, we give an overview of the most recent developments and applications in 3D printing of MEMS.
Multiple Fungal Metabolites Including Mycotoxins in Naturally Infected and Fusarium-Inoculated Wheat Samples
Authors: Spanic Valentina , Katanic Zorana , Sulyok Michael , Krska Rudolf , Puskas Katalin , Vida Gyula , Drezner Georg , Šarkanj Bojan
Published: April 2020

In this study, the occurrence of multiple fungal metabolites including mycotoxins was determined in four different winter wheat varieties in a field experiment in Croatia. One group was naturally infected, while the second group was inoculated with a Fusarium graminearum and F. culmorum mixture to simulate a worst-case infection scenario. Data on the multiple fungal metabolites including mycotoxins were acquired with liquid chromatography with mass spectrometry (LC-MS/MS) multi-(myco)toxin method. In total, 36 different fungal metabolites were quantified in this study: the Fusarium mycotoxins deoxynivalenol (DON), DON-3-glucoside (D3G), 3-acetyldeoxynivalenol (3-ADON), culmorin (CULM), 15-hydroxyculmorin, 5-hydroxyculmorin, aurofusarin, rubrofusarin, enniatin (Enn) A, Enn A1, Enn B, Enn B1, Enn B2, Enn B3, fumonisin B1, fumonisin B2, chrysogin, zearalenone (ZEN), moniliformin (MON), nivalenol (NIV), siccanol, equisetin, beauvericin (BEA), and antibiotic Y; the Alternaria mycotoxins alternariol, alternariolmethylether, altersetin, infectopyron, tentoxin, tenuazonic acid; the Aspergillus mycotoxin kojic acid; unspecific metabolites butenolid, brevianamid F, cyclo(L-Pro-L-Tyr), cyclo(L-Pro-L-Val), and tryptophol. The most abundant mycotoxins in the inoculated and naturally contaminated samples, respectively, were found to occur at the following average concentrations: DON (19,122/1504 µg/kg), CULM (6109/1010 µg/kg), 15-hydroxyculmorin (56,022/1301 µg/kg), 5-hydroxyculmorin (21,219/863 µg/kg), aurofusarin (43,496/1266 µg/kg). Compared to naturally-infected samples, Fusarium inoculations at the flowering stage increased the concentrations of all Fusarium mycotoxins, except enniatins and siccanol in Ficko, the Aspergillus metabolite kojic acid, the Alternaria mycotoxin altersetin, and unspecific metabolites brevianamid F, butenolid, cyclo(L-Pro-L-Tyr), and cyclo(L-Pro-L-Val). In contrast to these findings, because of possible antagonistic actions, Fusarium inoculation decreased the concentrations of the Alternaria toxins alternariol, alternariolmethylether, infectopyron, tentoxin, tenuazonic acid, as well as the concentration of the nonspecific metabolite tryptophol.
Human Antimicrobial Peptide Hepcidin 25-Induced Apoptosis in Candida albicans
Authors: Chen Ruei-Ching , Lan Chung-Yu
Published: April 2020

Hepcidin 25 (hep 25) is a cysteine-rich 25-amino acid antimicrobial peptide containing the amino-terminal Cu(II)/Ni(II)-binding (ATCUN) motif. Upon metal binding, the ATCUN motif is known to be involved in the generation of reactive oxygen species (ROS), especially hydrogen peroxide and hydroxyl radicals, which act against different bacterial species. However, the antifungal activity and its correlation to the Cu(II)-ATCUN complex of Hep 25 are still poorly understood. Here, we found that ROS accumulation plays an important role in the fungicidal activity of hep 25 against Candida albicans. In addition, Annexin V-FITC staining and TUNEL assay results provide clues about the apoptosis induced by hep 25. Moreover, hep 25 also increases the generation of ROS, possibly because of copper binding to the ATCUN motif, which is relevant to its activity against C. albicans. Finally, the C. albicans killing action of hep 25 is an energy- and temperature-dependent process that does not involve targeting the membrane. Taken together, our results provide new insights into the mechanisms of hep 25 against C. albicans cells and the potential use of hep 25 and its derivatives as novel antifungal agents.
Investigation of the Causes of Shigatoxigenic Escherichia coli PCR Positive and Culture Negative Samples
Authors: Macori Guerrino , McCarthy Siobhán C. , Burgess Catherine M. , Fanning Séamus , Duffy Geraldine
Published: April 2020

Molecular methods may reveal the presence of pathogens in samples through the detection of specific target gene(s) associated with microorganisms, but often, the subsequent cultural isolation of the pathogen is not possible. This discrepancy may be related to low concentration of the cells, presence of dead cells, competitive microflora, injured cells and cells in a viable but non-culturable state, free DNA and the presence of free bacteriophages which can carry the target gene causing the PCR-positive/culture-negative results. Shiga-toxigenic Escherichia coli (STEC) was used as a model for studying this phenomenon, based on the phage-encoded cytotoxins genes (Stx family) as the detection target in samples through real-time qPCR. Stx phages can be integrated in the STEC chromosome or can be isolated as free particles in the environment. In this study, a combination of PCR with culturing was used for investigating the presence of the stx1 and stx2 genes in 155 ovine recto-anal junction swab samples (method (a)-PCR). Samples which were PCR-positive and culture-negative were subjected to additional analyses including detection of dead STEC cells (method (b)-PCR-PMA dye assay), presence of Stx phages (method (c)-plaque assays) and inducible integrated phages (method (d)-phage induction). Method (a) showed that even though 121 samples gave a PCR-positive result (78%), only 68 samples yielded a culturable isolate (43.9%). Among the 53 (34.2%) PCR-positive/culture-negative samples, 21 (39.6%) samples were shown to have STEC dead cells only, eight (15.1%) had a combination of dead cells and inducible stx phage, while two samples (3.8%) had a combination of dead cells, inducible phage and free stx phage, and a further two samples had Stx1 free phages only (3.8%). It was thus possible to reduce the samples with no explanation to 20 (37.7% of 53 samples), representing a further step towards an improved understanding of the STEC PCR-positive/culture-negative phenomenon.
Autochthonous Cases of Mucosal Leishmaniasis in Northeastern Italy: Clinical Management and Novel Treatment Approaches
Authors: Gaspari Valeria , Zaghi Irene , Macrì Giovanni , Patrizi Annalisa , Salfi Nunzio , Locatelli Francesca , Carra Elena , Re Maria Carla , Varani Stefania
Published: April 2020

Mucosal leishmaniasis (ML) is a rare clinical variant of tegumentary leishmaniasis in Mediterranean Europe. Here we report on three autochthonous cases of head and neck ML in patients living in Northeastern Italy. Patients presented with non-specific, long-standing symptoms of upper respiratory tract involvement, mimicking other diseases. Parasitological diagnosis was reached by histopathology, immunohistochemistry and molecular biology on tissue specimens. Leishmania infantum was identified by molecular typing in all three cases. All patients reached a complete remission with protracted multivalent antileishmanial drugs; in one case, a novel approach of combined medical and endoscopic surgical treatment was carried out. High clinical suspicion led to a prompt diagnosis and deployment of a multivalent treatment. ML should be considered in the differential diagnosis of nasal, oral, and pharyngolaryngeal lesions in endemic areas. A prompt diagnosis is mandatory to establish a correct management; different antileishmanial medications as well as endoscopic surgical options may be required to reach a complete remission.
Ferritin: An Inflammatory Player Keeping Iron at the Core of Pathogen-Host Interactions
Authors: Moreira Ana C. , Mesquita Gonçalo , Gomes Maria Salomé
Published: April 2020

Iron is an essential element for virtually all cell types due to its role in energy metabolism, nucleic acid synthesis and cell proliferation. Nevertheless, if free, iron induces cellular and organ damage through the formation of free radicals. Thus, iron levels must be firmly controlled. During infection, both host and microbe need to access iron and avoid its toxicity. Alterations in serum and cellular iron have been reported as important markers of pathology. In this regard, ferritin, first discovered as an iron storage protein, has emerged as a biomarker not only in iron-related disorders but also in inflammatory diseases, or diseases in which inflammation has a central role such as cancer, neurodegeneration or infection. The basic research on ferritin identification and functions, as well as its role in diseases with an inflammatory component and its potential as a target in host-directed therapies, are the main considerations of this review.
Forest Tree Associated Bacterial Diffusible and Volatile Organic Compounds against Various Phytopathogenic Fungi
Authors: Kong Wei-Liang , Li Pu-Sheng , Wu Xiao-Qin , Wu Tian-Yu , Sun Xiao-Rui
Published: April 2020

Plant growth-promoting rhizobacteria (PGPR) can potentially be used as an alternative strategy to control plant diseases. In this study, strain ST–TJ4 isolated from the rhizosphere soil of a healthy poplar was found to have a strong antifungal activity against 11 phytopathogenic fungi in agriculture and forestry. Strain ST–TJ4 was identified as Pseudomonas sp. based on 16S rRNA-encoding gene sequences. The bacterium can produce siderophores, cellulase, and protease, and has genes involved in the synthesis of phenazine, 1–phenazinecarboxylic acid, pyrrolnitrin, and hydrogen cyanide. Additionally, the volatile compounds released by strain ST–TJ4 can inhibit the mycelial growth of plant pathogenic fungi more than diffusible substances can. Based on volatile compound profiles of strain ST–TJ4 obtained from headspace collection and GC–MS/MS analysis, 1-undecene was identified. In summary, the results suggested that Pseudomonas sp. ST–TJ4 can be used as a biocontrol agent for various plant diseases caused by phytopathogenic fungi.
Epidemiology and Genetic Variability of Circulating Influenza B Viruses in Uruguay, 2012–2019
Authors: Rivas María José , Alegretti Miguel , Cóppola Leticia , Ramas Viviana , Chiparelli Héctor , Goñi Natalia
Published: April 2020

Influenza B viruses (IBV) are an important cause of morbidity and mortality during interpandemic periods in the human population. Two phylogenetically distinct IBV lineages, B/Yamagata and B/Victoria, co-circulate worldwide and they present challenges for vaccine strain selection. Until the present study, there was little information regarding the pattern of the circulating strains of IBV in Uruguay. A subset of positive influenza B samples from influenza-like illness (ILI) outpatients and severe acute respiratory illness (SARI) inpatients detected in sentinel hospitals in Uruguay during 2012–2019 were selected. The sequencing of the hemagglutinin (HA) and neuraminidase (NA) genes showed substitutions at the amino acid level. Phylogenetic analysis reveals the co-circulation of both lineages in almost all seasonal epidemics in Uruguay, and allows recognizing a lineage-level vaccine mismatch in approximately one-third of the seasons studied. The epidemiological results show that the proportion of IBV found in ILI was significantly higher than the observed in SARI cases across different groups of age (9.7% ILI, 3.2% SARI) and patients between 5–14 years constituted the majority (33%) of all influenza B infection (p < 0.05). Interestingly, we found that individuals >25 years were particularly vulnerable to Yamagata lineage infections.
Biodegradation of Allethrin by a Novel Fungus Fusarium proliferatum Strain CF2, Isolated from Contaminated Soils
Authors: Bhatt Pankaj , Zhang Wenping , Lin Ziqiu , Pang Shimei , Huang Yaohua , Chen Shaohua
Published: April 2020

Continuous use of allethrin has resulted in heavy environmental contamination and has raised public concern about its impact on human health, yet little is known about the kinetics and microbial degradation of this pesticide. This study reported the degradation kinetics in a novel fungal strain, Fusarium proliferatum CF2, isolated from contaminated agricultural fields. Strain CF2 utilized 50 mg·L−1 of allethrin as the sole carbon source for growth in minimal salt medium and tolerated high concentrations of allethrin of up to 1000 mg·L−1. The optimum degradation conditions for strain CF2 were determined to be a temperature of 26 °C and pH 6.0 using response surface methodology. Under optimum conditions, strain CF2 completely degraded allethrin within 144 hours. The degradation kinetics of allethrin followed first order reaction kinetics. Kinetics analysis showed that its half-life was substantially reduced by 507.1 hours, as compared to the uninoculated control. This study provides new insights into the microbial degradation of allethrin with fungal F. proliferatum CF2.
Pathogenesis of Uveitis in Ebola Virus Disease Survivors: Evolving Understanding from Outbreaks to Animal Models
Authors: Hartley Caleb , Bavinger J. Clay , Kuthyar Sanjana , Shantha Jessica G. , Yeh Steven
Published: April 2020

Ebola virus disease (EVD) and emerging infectious disease threats continue to threaten life, prosperity and global health security. To properly counteract EVD, an improved understanding of the long-term impact of recent EVD outbreaks in West Africa and the Democratic Republic of Congo are needed. In the wake of recent outbreaks, numerous health sequelae were identified in EVD survivors. These findings include joint pains, headaches, myalgias, and uveitis, a vision-threatening inflammatory condition of the eye. Retrospective and more recent prospective studies of EVD survivors from West Africa have demonstrated that uveitis may occur in 13–34% of patients with an increase in prevalence from baseline to 12-month follow-up. The clinical spectrum of disease ranges from mild, anterior uveitis to severe, sight-threatening panuveitis. Untreated inflammation may ultimately lead to secondary complications of cataract and posterior synechiae, with resultant vision impairment. The identification of Ebola virus persistence in immune privileged organs, such as the eye, with subsequent tissue inflammation and edema may lead to vision loss. Non-human primate models of EVD have demonstrated tissue localization to the eye including macrophage reservoirs within the vitreous matter. Moreover, in vitro models of Ebola virus have shown permissiveness in retinal pigment epithelial cells, potentially contributing to viral persistence. Broad perspectives from epidemiologic studies of the outbreak, animal modeling, and immunologic studies of EVD survivors have demonstrated the spectrum of the eye disease, tissue specificity of Ebola virus infection, and antigen-specific immunologic response. Further studies in these areas will elucidate the mechanisms of this highly prevalent disease with the potential for improved therapeutics for Ebola virus in immune-privileged sites.
Emergence of a Multidrug-Resistant Enterobacter hormaechei Clinical Isolate from Egypt Co-Harboring mcr-9 and blaVIM-4
Authors: Soliman Ahmed M. , Maruyama Fumito , Zarad Hoda O. , Ota Atsushi , Nariya Hirofumi , Shimamoto Toshi , Shimamoto Tadashi
Published: April 2020

This study describes the first full genomic sequence of an mcr-9 and blaVIM-4-carrying multidrug-resistant Enterobacter hormaechei clinical isolate from Egypt. The strain was isolated in April 2015 from the sputum of a patient in Cairo, Egypt. The mcr-9 and blaVIM-4 genes were identified by PCR screening and DNA sequencing; the isolate was subjected to antimicrobial susceptibility testing, conjugation experiments, and whole genomic sequencing. mcr-9 and blaVIM-4 were carried by an IncHI2 plasmid, pAMS-38a (281,121 bp in size); the plasmid also carried genes conferring resistance against sulfonamides (sul1), quinolones (qnrA1), trimethoprim (dfrA1), β-lactams (blaTEM-1B), aminoglycosides (aac (6’)-II, aadA23, aadA2b, and ant(2’’)-Ia). The strain was susceptible to colistin (MIC, <0.25 μg/mL); this could be due to the absence of the qseC/qseB regulatory system located downstream of mcr-9 in Enterobacterales, which is involved in the induction of colistin-resistance. The genetic context of mcr-9 and blaVIM-4 was identified as IS1-mcr-9-IS903-pcoS-∆pcoE-rcnA and intI1-blaVIM-4—aac (6’)-II-dfrA1-∆aadA23-smr-ISPa21-qacE∆1, respectively. This is the first report of an mcr-9 and blaVIM-4 /IncHI2-carrying multidrug-resistant E. hormaechei clinical isolate from Africa and the Middle East. Plasmids of the IncHI2 group and the two insertion sequences (IS1, and IS903) might be the main vehicles for dissemination of mcr-9. Further screening for mcr-9 is essential for identifying its incidence and to prevent its dissemination.
Experimental Infection of Dogs with Toscana Virus and Sandfly Fever Sicilian Virus to Determine Their Potential as Possible Vertebrate Hosts
Authors: Muñoz Clara , Ayhan Nazli , Ortuño Maria , Ortiz Juana , Gould Ernest A. , Maia Carla , Berriatua Eduardo , Charrel Remi N.
Published: April 2020

The sandfly-borne Toscana phlebovirus (TOSV), a close relative of the sandfly fever Sicilian phlebovirus (SFSV), is one of the most common causes of acute meningitis or meningoencephalitis in humans in the Mediterranean Basin. However, most of human phlebovirus infections in endemic areas either are asymptomatic or cause mild influenza-like illness. To date, a vertebrate reservoir for sandfly-borne phleboviruses has not been identified. Dogs are a prime target for blood-feeding phlebotomines and are the primary reservoir of human sandfly-borne Leishmania infantum. However, there are no definitive studies to assess whether dogs play a significant role as a reservoir host for human phlebovirus survival in the environment. Here, we have evaluated the susceptibility of domestic dogs to infection by TOSV and SFSV following the direct inoculation of the infectious virus. After experimental infection, the presence of viral RNA was investigated in plasma, urine, saliva, conjunctiva, faeces, semen, and bone marrow samples from 0 to 91 days postinoculation (dpi), as well as in plasma, saliva, and tears samples at 760 dpi. None of the challenged dogs developed clinical signs of infection with either TOSV or SFSV. SFSV RNA was never detected. TOSV RNA was not in any of the specimen types, except for plasma samples that showed low viral loads, although irregularly. None of the dogs developed detectable neutralizing antibodies after a single challenge dose of either TOSV or SFSV. However, a second challenge dose of virus given 56 days later elicited neutralizing antibodies, implying that the first inoculation of virus primed the animals for an anamnestic response following the second challenge. These results demonstrated that healthy domestic dogs are not highly susceptible to infection by TOSV or SFSV and do not develop significant viremia or excrete virus following infection. Consequently, dogs are unlikely natural reservoir hosts of infection and do not appear to play a significant role in phlebovirus transmission cycles.
FDA-Approved Drugs Efavirenz, Tipranavir, and Dasabuvir Inhibit Replication of Multiple Flaviviruses in Vero Cells
Authors: Stefanik Michal , Valdes James J. , Ezebuo Fortunatus C. , Haviernik Jan , Uzochukwu Ikemefuna C. , Fojtikova Martina , Salat Jiri , Eyer Ludek , Ruzek Daniel
Published: April 2020

Vector-borne flaviviruses (VBFs) affect human health worldwide, but no approved drugs are available specifically to treat VBF-associated infections. Here, we performed in silico screening of a library of U.S. Food and Drug Administration-approved antiviral drugs for their interaction with Zika virus proteins. Twelve hit drugs were identified by the docking experiments and tested in cell-based antiviral assay systems. Efavirenz, tipranavir, and dasabuvir at micromolar concentrations were identified to inhibit all VBFs tested; i.e., two representatives of mosquito-borne flaviviruses (Zika and West Nile viruses) and one representative of flaviviruses transmitted by ticks (tick-borne encephalitis virus). The results warrant further research into these drugs, either individually or in combination, as possible pan-flavivirus inhibitors.
Editorial for Special Issue “Heavy Minerals”
Authors: Andò Sergio
Published: April 2020

This special volume, published 13 years after the monumental volume “Heavy Minerals in Use” edited by Maria Mange and David Wright, demonstrates that the use of heavy minerals as provenance tracers is alive and in full health [...]
Native Seedling Colonization on Stockpiled Mine Soils Is Constrained by Site Conditions and Competition with Exotic Species
Authors: Baethke Kathy A. , Ploughe Laura W. , Gardner Wendy C. , Fraser Lauchlan H.
Published: April 2020

Sites disturbed through mining practices can be challenging to restore with native vegetation, as the path of ecological succession is often unknown and hard to predict. We conducted an establishment study that explored restoration strategies to increase native vegetation on a newly formed stockpile of soil at New Gold’s New Afton Mine in British Columbia, Canada. Establishment of native species in semi-arid grasslands is often constrained by seed and seedling microsite limitations, so treatments were imposed to assist in reducing these limitations. We established a fully factorial design with a native seed treatment (seeded and unseeded) and four soil preparations: raking, hydroseed slurry, a combination of raking and hydroseed slurry, and no preparation. Raking assisted in increasing total seedling establishment, regardless of whether the site was seeded or not. Raking and seeding increased the number of native seedlings and resulted in the greatest species richness, suggesting that native seedling establishment is primarily seed-limited, but that microclimate is also important for the establishment of some native seeds. We found that exotic species were able to capitalize on the disturbance and outcompeted the native species, but reducing seed-limitations by sowing more native seeds and increasing available microclimates by raking or tilling may increase native species’ success at the establishment phase.
Assessment of Water Quality and Soil Salinity in the Agricultural Coastal Plain (Ravenna, North Italy)
Authors: Vittori Antisari Livia , Speranza Maria , Ferronato Chiara , De Feudis Mauro , Vianello Gilmo , Falsone Gloria
Published: April 2020

To improve knowledge on salt leaching suitability on different soils, in Arenosols and Cambisols croplands in the coastal area of Ravenna (Italy), soil samples were collected in the non-irrigation winter period and irrigation summer period. Concurrently, waters of the canal network were also investigated. Soil samples were analyzed for pH, carbonate, total organic carbon (TOC), particle size distribution, electrical conductivity (EC), bulk density (BD) and water content at field capacity (FC). Water samples were investigated for pH, EC, biological and chemical oxygen demand, sodium adsorption ratio, phosphorus, nitrogen, sulfates and chlorides. All soils had low TOC concentrations and Arenosols showed the lowest clay content, BD and FC. Soils had similar EC values in winter, but in summer the lowest ones were observed in Arenosols, suggesting that irrigation mitigated salinization in Arenosols, while the high clay content, BD and FC prevented or limited the salt leaching in Cambisols. In summer, the increase of total nitrogen and biological oxygen demand, especially in drainage channels, might suggest the leaching of soluble nutrients and organic matter from soils due to the high irrigation water volumes. Finally, our findings stress the need to consider soil type and properties to contrast soil salinization without negative effects on soil C leaching caused by salt leaching practice.
Zoned Laurite from the Merensky Reef, Bushveld Complex, South Africa: “Hydrothermal” in Origin?
Authors: Zaccarini Federica , Garuti Giorgio
Published: April 2020

Laurite, ideally (Ru,Os)S2, is a common accessory mineral in podiform and stratiform chromitites and, to a lesser extent, it also occurs in placer deposits and is associated with Ni-Cu magmatic sulfides. In this paper, we report on the occurrence of zoned laurite found in the Merensky Reef of the Bushveld layered intrusion, South Africa. The zoned laurite forms relatively large crystals of up to more than 100 µm, and occurs in contact between serpentine and sulfides, such as pyrrhotite, chalcopyrite, and pentlandite, that contain small phases containing Pb and Cl. Some zoned crystals of laurite show a slight enrichment in Os in the rim, as typical of laurite that crystallized at magmatic stage, under decreasing temperature and increasing sulfur fugacity, in a thermal range of about 1300–1000 °C. However, most of the laurite from the Merensky Reef are characterized by an unusual zoning that involves local enrichment of As, Pt, Ir, and Fe. Comparison in terms of Ru-Os-Ir of the Merensky Reef zoned laurite with those found in the layered chromitites of the Bushveld and podiform chromitites reveals that they are enriched in Ir. The Merensky Reef zoned laurite also contain high amount of As (up to 9.72 wt%), Pt (up to 9.72 wt%) and Fe (up to 14.19 wt%). On the basis of its textural position, composition, and zoning, we can suggest that the zoned laurite of the Merensky Reef is “hydrothermal” in origin, having crystallized in the presence of a Cl- and As-rich hydrous solution, at temperatures much lower than those typical of the precipitation of magmatic laurite. Although, it remains to be seen whether the “hydrothermal” laurite precipitated directly from the hydrothermal fluid, or it represents the alteration product of a pre-existing laurite reacting with the hydrothermal solution.
Characterization of Ginkgo biloba Leaf Flavonoids as Neuroexocytosis Regulators
Authors: Ban Choongjin , Park Joon-Bum , Cho Sora , Kim Hye Rin , Kim Yong Joon , Bae Hyungjin , Kim Chinhan , Kang Hakhee , Jang Davin , Shin Yong Sub , Kim Dae-Ok , Kim Hyunggun , Kweon Dae-Hyuk
Published: April 2020

Ginkgo biloba leaf (GBL) is known as a potential source of bioactive flavonoids, such as quercetin, arresting the neuronal soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE)-zippering. Here, the GBL flavonoids were isolated in two different manners and then examined for their bioactivity, physicochemical stability, and biocompatibility. The majority of flavonoids in the non-hydrolyzed and acidolyzed isolates, termed non-hydrolyzed isolate (NI) and acidolyzed isolate (AI) hereafter, were rich in flavonol glycosides and aglycones, respectively. Glycosidic/aglyconic quercetin and kaempferol were abundant in both NI and AI, whereas a little of apigenin, luteolin, and isorhamnetin were found in AI. NI was more thermostable in all pH ranges than quercetin, kaempferol, and AI. NI and AI both inhibited neurotransmitter release from differentiated neuronal PC-12 cells. NI and AI showed 1/2–1/3 lower EC50/CC50 values than quercetin and kaempferol. The NI and AI exhibited no toxicity assessed by the tests on chorioallantoic membranes of hen’s eggs, removing toxicological concerns of irritation potential. Moreover, GBL isolates, particularly AI, showed antioxidant and anti-inflammatory activities in the use below the CC50 levels. Taken together, these results suggest that GBL isolates that are rich in antioxidant flavonoids are effective anti-neuroexocytotic agents with high stability and low toxicity.
Sensing Precursors of Illegal Drugs—Rapid Detection of Acetic Anhydride Vapors at Trace Levels Using Photoionization Detection and Ion Mobility Spectrometry
Authors: Bocos-Bintintan Victor , Ghira George-Bogdan , Anton Mircea , Martiniuc Aurel-Vasile , Ratiu Ileana-Andreea
Published: April 2020

Sensitive real-time detection of vapors produced by the precursors, reagents and solvents used in the illegal drugs manufacture represents a priority nowadays. Acetic anhydride (AA) is the key chemical used as acetylation agent in producing the illegal drugs heroin and methaqualone. This study was directed towards quick detection and quantification of AA in air, using two fast and very sensitive analytical techniques: photoionization detection (PID) and ion mobility spectrometry (IMS). Results obtained indicated that both PID and IMS can sense AA at ultra-trace levels in air, but while PID produces a non-selective response, IMS offers richer information. Ion mobility spectrometric response in the positive ion mode presented one product ion, at reduced ion mobility K0 of 1.89 cm2 V−1 s−1 (almost overlapped with positive reactant ion peak), while in the negative ion mode two well separated product ions, with K0 of 1.90 and 1.71 cm2 V−1 s−1, were noticed. Our study showed that by using a portable, commercial IMS system (model Mini IMS, I.U.T. GmbH Berlin) AA can be easily measured at concentrations of 0.05 ppmv (0.2 mg m−3) in negative ion mode. Best selectivity and sensitivity of the IMS response were therefore achieved in the negative operation mode.
Evaluation of the Cytotoxic Activity of the Usnea barbata (L.) F. H. Wigg Dry Extract
Authors: Popovici Violeta , Bucur Laura Adriana , Schröder Verginica , Gherghel Daniela , Mihai Cosmin Teodor , Caraiane Aureliana , Badea Florin Ciprian , Vochița Gabriela , Badea Victoria
Published: April 2020

The secondary metabolites of lichens have proven to be promising sources of anticancer drugs; one of the most important of these is usnic acid, which is a phenolic compound with dibenzofuran structure that is responsible for the numerous biological actions of lichens of genus Usnea. As a result, in this study, we related to this phenolic secondary metabolite. The aim of the present study is the evaluation of the cytotoxic activity of Usnea barbata (L.) F. H. Wigg dry acetone extract (UBE). In advance, the usnic acid content was determined in various extracts of Usnea barbata (L.) F. H. Wigg: the liquid extracts were found in water, ethanol, acetone, and the dry acetone extract; the highest usnic acid quantity was found in the dry acetone extract. First, the cytotoxic action of UBE was assessed using Brine Shrimp Lethality (BSL) test; a significant lethal effect was obtained after 24 h of treatment at high used concentrations of UBE, and it was quantified by the high mortality rate of the Artemia salina (L.) larvae. Secondly, in vitro cytotoxicity of UBE was evaluated on human tongue squamous cells carcinoma, using CAL 27 (ATCC® CRL-2095™) cell line. The most intense cytotoxic effect of UBE on CAL 27 cells was registered after 24 h; this response is directly proportional with the tested UBE concentrations. The obtained results have been reported regarding usnic acid content of UBE, and the data show that CAL 27 cells death was induced by apoptosis and high oxidative stress.
Rapid Raman Spectroscopic Analysis of Stress Induced Degradation of the Pharmaceutical Drug Tetracycline
Authors: Domes Christian , Frosch Timea , Popp Juergen , Frosch Torsten
Published: April 2020

Stress factors caused by inadequate storage can induce the unwanted degradation of active compounds in pharmaceutical formulations. Resonance Raman spectroscopy is presented as an analytical tool for rapid monitoring of small concentration changes of tetracycline and the metabolite 4˗epianhydrotetracycline. These degradation processes were experimentally induced by changes in temperature, humidity, and irradiation with visible light over a time period of up to 23 days. The excitation wavelength λexc = 413 nm was proven to provide short acquisition times for the simultaneous Raman spectroscopic detection of the degradation of tetracycline and production of its impurity in small sample volumes. Small concentration changes could be detected (down to 1.4% for tetracycline and 0.3% for 4-epianhydrotetracycline), which shows the potential of resonance Raman spectroscopy for analyzing the decomposition of pharmaceutical products.
Synthesis, Characterization and Biological Activity of Novel Cu(II) Complexes of 6-Methyl-2-Oxo-1,2-Dihydroquinoline-3-Carbaldehyde-4n-Substituted Thiosemicarbazones
Authors: Ramachandran Eswaran , Gandin Valentina , Bertani Roberta , Sgarbossa Paolo , Natarajan Karuppannan , Bhuvanesh Nattamai S. P. , Venzo Alfonso , Zoleo Alfonso , Mozzon Mirto , Dolmella Alessandro , Albinati Alberto , Castellano Carlo , Reis Conceição Nuno , C. Guedes da Silva M. Fátima , Marzano Cristina
Published: April 2020

Three new 6-methyl-2-oxo-1,2-dihydroquinoline-3-carbaldehyde-thiosemicarbazones-N-4-substituted pro-ligands and their Cu(II) complexes (1, -NH2; 2, -NHMe; 3, -NHEt) have been prepared and characterized. In both the X-ray structures of 1 and 3, two crystallographically independent complex molecules were found that differ either in the nature of weakly metal-binding species (water in 1a and nitrate in 1b) or in the co-ligand (water in 3a and methanol in 3b). Electron Paramagnetic Resonance (EPR) measurements carried out on complexes 1 and 3 confirmed the presence of such different species in the solution. The electrochemical behavior of the pro-ligands and of the complexes was investigated, as well as their biological activity. Complexes 2 and 3 exhibited a high cytotoxicity against human tumor cells and 3D spheroids derived from solid tumors, related to the high cellular uptake. Complexes 2 and 3 also showed a high selectivity towards cancerous cell lines with respect to non-cancerous cell lines and were able to circumvent cisplatin resistance. Via the Transmission Electron Microscopy (TEM) imaging technique, preliminary insights into the biological activity of copper complexes were obtained.
Molecular Crystallization Inhibitors for Salt Damage Control in Porous Materials: An Overview
Authors: Bracciale Maria Paola , Sammut Svetlana , Cassar JoAnn , Santarelli Maria Laura , Marrocchi Assunta
Published: April 2020

The use of inhibition chemicals holds the prospect of an efficient strategy to control crystallization in porous materials, thereby potentially contributing to the prevention or mitigation of the salt decay phenomenon in modern as well as historical building materials in a more sustainable manner. In this review, we first provide an essential background on the mechanism of salt crystallization and on the factors influencing this phenomenon; next, we illustrate the mechanism at the basis of the action of crystal growth inhibitors, and critically discuss the major advances in the development of different families of inhibitors, particularly focusing on their influence on salt transport and crystallization within the structure of porous media. Specifically, correlations between the crystallization inhibition processes in porous materials and variables, such as porous substrate composition and properties, contaminant salt type and concentrations, microclimatic conditions, inhibiting solution concentration and properties, and application methods, will be highlighted. Environmental aspects, limitations, and problems associated with some inhibition chemicals are also taken into account. Finally, a survey and a discussion on the most representative experimental techniques and instrumentation available to assess qualitatively and quantitatively the inhibitor effectiveness, as well as recently developed modelling tools are given out.
Advances on Antiviral Activity of Morus spp. Plant Extracts: Human Coronavirus and Virus-Related Respiratory Tract Infections in the Spotlight
Authors: Thabti Inès , Albert Quentin , Philippot Stéphanie , Dupire François , Westerhuis Brenda , Fontanay Stéphane , Risler Arnaud , Kassab Thomas , Elfalleh Walid , Aferchichi Ali , Varbanov Mihayl
Published: April 2020

(1) Background: Viral respiratory infections cause life-threatening diseases in millions of people worldwide every year. Human coronavirus and several picornaviruses are responsible for worldwide epidemic outbreaks, thus representing a heavy burden to their hosts. In the absence of specific treatments for human viral infections, natural products offer an alternative in terms of innovative drug therapies. (2) Methods: We analyzed the antiviral properties of the leaves and stem bark of the mulberry tree (Morus spp.). We compared the antiviral activity of Morus spp. on enveloped and nonenveloped viral pathogens, such as human coronavirus (HCoV 229E) and different members of the Picornaviridae family—human poliovirus 1, human parechovirus 1 and 3, and human echovirus 11. The antiviral activity of 12 water and water–alcohol plant extracts of the leaves and stem bark of three different species of mulberry—Morus alba var. alba, Morus alba var. rosa, and Morus rubra—were evaluated. We also evaluated the antiviral activities of kuwanon G against HCoV-229E. (3) Results: Our results showed that several extracts reduced the viral titer and cytopathogenic effects (CPE). Leaves’ water-alcohol extracts exhibited maximum antiviral activity on human coronavirus, while stem bark and leaves’ water and water-alcohol extracts were the most effective on picornaviruses. (4) Conclusions: The analysis of the antiviral activities of Morus spp. offer promising applications in antiviral strategies.
Insights on Ultrafiltration-Based Separation for the Purification and Quantification of Methotrexate in Nanocarriers
Authors: Marques Sara S. , Ramos Inês I. , Fernandes Sara R. , Barreiros Luisa , Lima Sofia A. C. , Reis Salette , Domingues M. Rosário M. , Segundo Marcela A.
Published: April 2020

The evaluation of encapsulation efficiency is a regulatory requirement for the characterization of drug delivery systems. However, the difficulties in efficiently separating nanomedicines from the free drug may compromise the achievement of accurate determinations. Herein, ultrafiltration was exploited as a separative strategy towards the evaluation of methotrexate (MTX) encapsulation efficiency in nanostructured lipid carriers and polymeric nanoparticles. The effect of experimental conditions such as pH and the amount of surfactant present in the ultrafiltration media was addressed aiming at the selection of suitable conditions for the effective purification of nanocarriers. MTX-loaded nanoparticles were then submitted to ultrafiltration and the portions remaining in the upper compartment of the filtering device and in the ultrafiltrate were collected and analyzed by HPLC-UV using a reversed-phase (C18) monolithic column. A short centrifugation time (5 min) was suitable for establishing the amount of encapsulated MTX in nanostructured lipid carriers, based on the assumption that the free MTX concentration was the same in the upper compartment and in the ultrafiltrate. The defined conditions allowed the efficient separation of nanocarriers from the free drug, with recoveries of >85% even when nanoparticles were present in cell culture media and in pig skin surrogate from permeation assays.
A Gram-Scale Limonene Production Process with Engineered Escherichia coli
Authors: Rolf Jascha , Julsing Mattijs K. , Rosenthal Katrin , Lütz Stephan
Published: April 2020

Monoterpenes, such as the cyclic terpene limonene, are valuable and important natural products widely used in food, cosmetics, household chemicals, and pharmaceutical applications. The biotechnological production of limonene with microorganisms may complement traditional plant extraction methods. For this purpose, the bioprocess needs to be stable and ought to show high titers and space-time yields. In this study, a limonene production process was developed with metabolically engineered Escherichia coli at the bioreactor scale. Therefore, fed-batch fermentations in minimal medium and in the presence of a non-toxic organic phase were carried out with E. coli BL21 (DE3) pJBEI-6410 harboring the optimized genes for the mevalonate pathway and the limonene synthase from Mentha spicata on a single plasmid. The feasibility of glycerol as the sole carbon source for cell growth and limonene synthesis was examined, and it was applied in an optimized fermentation setup. Titers on a gram-scale of up to 7.3 g·Lorg−1 (corresponding to 3.6 g·L−1 in the aqueous production phase) were achieved with industrially viable space-time yields of 0.15 g·L−1·h−1. These are the highest monoterpene concentrations obtained with a microorganism to date, and these findings provide the basis for the development of an economic and industrially relevant bioprocess.