Green synthesis of silver nanoparticles and biopolymer nanocomposites: a comparative study on physico-chemical, antimicrobial and anticancer activity

The current report was intended towards comparative study of green-synthesized biogenic Rhubarb silver nanoparticles (RS-AgNPs) and chitosan crosslinked silver nanocomposites (CSHD-AgNCs). The physico-chemical characterization was done by UV-visible, FTIR, scanning electron microscopy (SUM), transmission electron microscopy (TEM), TGA, XRD and zeta potential(zeta). The analysis and spectroscopic characterization was done by SEM and TEM and their results reveal that the nanoparticles are spherical in shape, with average size ranges from 5 to 50 nm, and was gathered by face centered cubic (FCC) structure throughout the polymer matrix and stable without any protecting or capping reagents over 450 days. The antimicrobial property of RS-AgNPs and CSHD-AgNCs (zeta = +29.6 and +32.8 mV) was evaluated against E. coli and S. aureus and showed an effective inhibitory property. The RS-AgNPs and CSHD-AgNCs were assessed for their anticancer activity against HeLa cell line by MTT method, and it reveals a dose response activity, time and cell line-dependent cytotoxicity. Based on the results obtained, the RS-AgNPs exhibited higher toxicity over CSHD-AgNCs after 24 h incubation of HeLa cells with different concentrations and is negligible for the aqueous Rhubarb extract. It was concluded that the changes in anticancer activity towards HeLa cells due to biological activity of silver nanoparticles depend on their method of biosynthesis and their physico-chemical nature.Ministry of Education, Youth and Sports of the Czech Republic-NPU Program I [LO1504]; Slovak Grant Agency VEGA [2/0124/18

Chitosan–silver nanocomposites: New functional biomaterial for health-care applications

Chitosan–silver nanocomposites (CS-HDA-AgNCs) was prepared using chitosan, biogenic silver nanocomposites, and crosslinker, hexamethylene 1,6-di(amino carboxysulfonate) (HDA). The film is flexible and transparent. Its physical, mechanical, thermal, hydrophilicity, and swelling properties were improved by HDA (2.5%). The antimicrobial activity of CS-HDA-AgNCs were not displayed any remarkable zone of inhibition but showed toxic effect in the presence of normal 3T3 fibroblasts and cancer HeLa cells. It decreases to ca. 5–7% for both cell lines. In conclusion, it can be mentioned that the CS-HDA-AgNCs, a kind of new functional biomaterial, could be useful for health-care applications. © 2018 Taylor & Francis.Ministry of Education, Youth and Sports of the Czech Republic - NPU Program I [LO1504]; Slovak Grant Agency VEGA [2/0156/15

Geographical and temporal distribution of SARS-CoV-2 clades in the WHO European Region, January to June 2020

We show the distribution of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) genetic clades over time and between countries and outline potential genomic surveillance objectives. We applied three genomic nomenclature systems to all sequence data from the World Health Organization European Region available until 10 July 2020. We highlight the importance of real-time sequencing and data dissemination in a pandemic situation, compare the nomenclatures and lay a foundation for future European genomic surveillance of SARS-CoV-2

Alimentary Infections by Tick-Borne Encephalitis Virus

Tick-borne encephalitis virus (TBEV) causes serious the neurological disease, tick-borne encephalitis (TBE). TBEV can be transmitted to humans by ticks as well as by the alimentary route, which is mediated through the consumption of raw milk products from infected ruminants such as sheep, goats, and cows. The alimentary route of TBEV was recognized in the early 1950s and many important experimental studies were performed shortly thereafter. Nowadays, alimentary TBEV infections are recognized as a relevant factor contributing to the overall increase in TBE incidences in Europe. This review aims to summarize the history and current extent of alimentary TBEV infections across Europe, to analyze experimental data on virus secretion in milk, and to review possible alimentary infection preventive measures

SARS-CoV-2 Exploits Non-Canonical Autophagic Processes to Replicate, Mature, and Egress the Infected Vero E6 Cells

The coronavirus transforms the cytoplasm of susceptible cells to support virus replication. It also activates autophagy-like processes, the role of which is not well understood. Here, we studied SARS-CoV-2-infected Vero E6 cells using transmission electron microscopy and autophagy PCR array. After 6–24 h post-infection (hpi), the cytoplasm of infected cells only contained double-membrane vesicles, phagophores, and phagosomes engulfing virus particles and cytoplasmic debris, including damaged mitochondria. The phagosomes interacted with the viral nucleoprotein complex, virus particles, mitochondria, and lipid droplets. The phagosomes transformed into egress vacuoles, which broke through the plasmalemma and discharged the virus particles. The Vero E6 cells exhibited pronounced virus replication at 6 hpi, which stabilized at 18–24 hpi at a high level. The autophagy PCR array tests revealed a significant upregulation of 10 and downregulation of 8 autophagic gene markers out of 84. Altogether, these results underline the importance of autophagy-like processes for SARS-CoV-2 maturation and egress, and point to deviations from a canonical autophagy response

Co-Circulation of West Nile, Usutu, and Tick-Borne Encephalitis Viruses in the Same Area: A Great Challenge for Diagnostic and Blood and Organ Safety

Viral infections caused by viruses from the family Flaviviridae such as Zika (ZIKV), Dengue (DENV), yellow fever (YFV), tick-borne encephalitis (TBEV), West Nile (WNV), and Usutu (USUV) are some of the most challenging diseases for recognition in clinical diagnostics and epidemiological tracking thanks to their short viremia, non-specific symptoms, and high cross-reactivity observed in laboratory techniques. In Central Europe, the most relevant endemic flaviviruses are mosquito-borne WNV and USUV, and tick-borne TBEV. All three viruses have been recognised to be responsible for human neuroinvasive diseases. Moreover, they are interrupting the blood and transplantation safety processes, when the great efforts made to save a patient’s life could be defeated by acquired infection from donors. Due to the trend of changing distribution and abundance of flaviviruses and their vectors influenced by global change, the co-circulation of WNV, USUV, and TBEV can be observed in the same area. In this perspective, we discuss the problems of flavivirus diagnostics and epidemiology monitoring in Slovakia as a model area of Central Europe, where co-circulation of WNV, USUV, and TBEV in the same zone has been recently detected. This new situation presents multiple challenges not only for diagnostics or surveillance but particularly also for blood and organ safety. We conclude that the current routinely used laboratory diagnostics and donor screening applied by the European Union (EU) regulations are out of date and the novel methods which have become available in recent years, e.g., next-gene sequencing or urine screening should be implemented immediately

Co-Circulation of West Nile, Usutu, and Tick-Borne Encephalitis Viruses in the Same Area: A Great Challenge for Diagnostic and Blood and Organ Safety

Viral infections caused by viruses from the family Flaviviridae such as Zika (ZIKV), Dengue (DENV), yellow fever (YFV), tick-borne encephalitis (TBEV), West Nile (WNV), and Usutu (USUV) are some of the most challenging diseases for recognition in clinical diagnostics and epidemiological tracking thanks to their short viremia, non-specific symptoms, and high cross-reactivity observed in laboratory techniques. In Central Europe, the most relevant endemic flaviviruses are mosquito-borne WNV and USUV, and tick-borne TBEV. All three viruses have been recognised to be responsible for human neuroinvasive diseases. Moreover, they are interrupting the blood and transplantation safety processes, when the great efforts made to save a patient’s life could be defeated by acquired infection from donors. Due to the trend of changing distribution and abundance of flaviviruses and their vectors influenced by global change, the co-circulation of WNV, USUV, and TBEV can be observed in the same area. In this perspective, we discuss the problems of flavivirus diagnostics and epidemiology monitoring in Slovakia as a model area of Central Europe, where co-circulation of WNV, USUV, and TBEV in the same zone has been recently detected. This new situation presents multiple challenges not only for diagnostics or surveillance but particularly also for blood and organ safety. We conclude that the current routinely used laboratory diagnostics and donor screening applied by the European Union (EU) regulations are out of date and the novel methods which have become available in recent years, e.g., next-gene sequencing or urine screening should be implemented immediately

Photoimmobilization of zwitterionic polymers on surfaces to reduce cell adhesion

Simple and robust methods for modifying hydrophobic polymer surfaces with zwitterionic polymers using UV irradiation were developed. Two random zwitterionic copolymers consisting of either carboxybetaine or sulfobetaine methacrylamide monomers and monomers bearing a photolabile azidophenyl group were directly photoimmobilized on polymeric surfaces (polyester, polyethylene and polystyrene) via covalent interactions in a spatially controlled manner. These copolymers were also electrospun to form self-standing mats. The modified surfaces were characterized by X-ray photoelectron spectroscopy, atomic force microscopy, infrared spectroscopy and contact angle measurements. The electrospinning method involved the use of a trifluoroethanol solution with a copolymer concentration in the range from 2 to 10 wt.%. BHK 21 cell adhesion to both modified surfaces and mats was dramatically reduced compared to unmodified surfaces.Scopu

Fitness of mCherry Reporter Tick-Borne Encephalitis Virus in Tick Experimental Models

The tick-borne encephalitis virus (TBEV) causes a most important viral life-threatening illness transmitted by ticks. The interactions between the virus and ticks are largely unexplored, indicating a lack of experimental tools and systematic studies. One such tool is recombinant reporter TBEV, offering antibody-free visualization to facilitate studies of transmission and interactions between a tick vector and a virus. In this paper, we utilized a recently developed recombinant TBEV expressing the reporter gene mCherry to study its fitness in various tick-derived in vitro cell cultures and live unfed nymphal Ixodes ricinus ticks. The reporter virus was successfully replicated in tick cell lines and live ticks as confirmed by the plaque assay and the mCherry-specific polymerase chain reaction (PCR). Although a strong mCherry signal determined by fluorescence microscopy was detected in several tick cell lines, the fluorescence of the reporter was not observed in the live ticks, corroborated also by immunoblotting. Our data indicate that the mCherry reporter TBEV might be an excellent tool for studying TBEV-tick interactions using a tick in vitro model. However, physiological attributes of a live tick, likely contributing to the inactivity of the reporter, warrant further development of reporter-tagged viruses to study TBEV in ticks in vivo

The Drug-Loaded Electrospun Poly(ε-Caprolactone) Mats for Therapeutic Application

Diclofenac sodium salt (DSS)-loaded electrospun nanofiber mats on the base of poly(ε-caprolactone) (PCL) were investigated as biocompatible nanofibrous mats for medical applications with the ability to inhibit bacterial infections. The paper presents the characteristics of fibrous mats made by electrospinning and determines the effect of medicament on the fiber morphology, chemical, mechanical and thermal properties, as well as wettability. PCL and DSS-loaded PCL nanofibrous mats were characterized using scanning electron microscopy, transmission electron microscopy, attenuated total reflectance-Fourier transform infrared spectrometry, dynamic mechanical analysis, and contact angle measurements. Electron paramagnetic resonance measurements confirmed the lifetime of DSS before and after application of high voltage during the electrospinning process. In vitro biocompatibility was studied, and it was proved to be of good viability with ~92% of the diploid human cells culture line composed of lung fibroblast (MRC 5) after 48 h of incubation. Moreover, the significant activity of DSS-loaded nanofibers against cancer cells, Ca Ski and HeLa, was established as well. It was shown that 12.5% (m/V) is the minimal concentration for antibacterial activity when more than 99% of Escherichia coli (Gram-negative) and 99% of Staphylococcus aureus (Gram-positive) have been exterminated