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Malwina Sosnowska,1 Mateusz Wierzbicki,1 Barbara Nasiłowska,2 Totka Nikolaeva Bakalova,3 Klara Piotrowska,4 Barbara Strojny-Cieślak,1 Ewa Sawosz,1 Marta Kutwin1 1Department of Nanobiotechnology, Institute of Biology, Warsaw University of Life Sciences, Warsaw, Poland; 2Biomedical Engineering Center, Institute of Optoelectronics, Military University of Technology, Warsaw, Poland; 3Department of Material Science, Faculty of Mechanical Engineering, Technical University of Liberec, Liberec, Czech Republic; 4Department of Animal Nutrition, Institute of Animal Sciences, Warsaw University of Life Sciences, Warsaw, PolandCorrespondence: Malwina Sosnowska, Department of Nanobiotechnology, Institute of Biology, Warsaw University of Life Sciences, st. Ciszewskiego 8, Warsaw, 02-786, Poland, Tel +48 22 593 66 71, Email malwina_sosnowska@sggw.edu.plIntroduction and Objective: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) enters the nasal cavity, penetrates the nasal epithelial cells through the interaction of its spike protein with the host cell receptor angiotensin-converting enzyme 2 (ACE2) and then triggers a cytokine storm. We aimed to assess the biocompatibility of fullerenol nanoparticles C60(OH)40 and ectoine, and to document their effect on the protection of primary human nasal epithelial cells (HNEpCs) against the effects of interaction with the fragment of virus - spike protein. This preliminary research is the first step towards the construction of a intranasal medical device with a protective, mechanical function against SARS-CoV-2 similar to that of personal protective equipment (eg masks).Methods: We used HNEpCs and the full-length spike protein from SARS-CoV-2 to mimic the first stage of virus infection. We assessed cell viability with the XTT assay and a spectrophotometer. May–Grünwald Giemsa and periodic acid–Schiff staining served to evaluate HNEpC morphology. We assessed reactive oxygen species (ROS) production by using 2′,7′-dichlorofluorescin diacetate and commercial kit. Finally, we employed reverse transcription polymerase chain reaction, Western blotting and confocal microscopy to determine the expression of angiotensin-converting enzyme 2 (ACE2) and inflammatory cytokines.Results: There was normal morphology and unchanged viability of HNEpCs after incubation with 10 mg/L C60(OH)40, 0.2% ectoine or their composite for 24 h. The spike protein exerted cytotoxicity via ROS production. Preincubation with the composite protected HNEpCs against the interaction between the spike protein and the host membrane and prevented the production of key cytokines characteristic of severe coronavirus disease 2019, including interleukin 6 and 8, monocyte chemotactic protein 1 and 2, tissue inhibitor of metalloproteinases 2 and macrophage colony-stimulating factor.Conclusion: In the future, the combination of fullerenol and ectoine may be used to prevent viral infections as an intranasal medical device for people with reduced immunity and damaged mucous membrane. Keywords: ACE2, cytokine storm, ectoine, nasal epithelium, polyhydroxylated fullerene, spike

Marta Kutwin,1 Malwina Sosnowska-Ławnicka,1 Barbara Nasiłowska,2 Agata Lange,1 Mateusz Wierzbicki,1 Sławomir Jaworski1 1Department of Nanobiotechnology, Institute of Biology, Warsaw University of Life Sciences, Warsaw, Poland; 2Institute of Optoelectronics, Military University of Technology, Warsaw, PolandCorrespondence: Marta Kutwin, Email marta_kutwin@sggw.edu.plPurpose: The use of nanotechnology in medicine has gained attention in developing drug delivery systems. GO has the potential to deliver microRNA (miRNA) mimics or antisense structures. MiRNAs regulate gene expression and their dysregulation is implicated in diseases, including cancer. This study aims to observe changes in morphology, viability, mRNA expression of mTOR/PI3K/Akt and PTEN genes in U87, U118, U251, A172 and T98 glioblastoma cells and xenograft models after GO self-assembly with mimic miRNA-7.Methods: Colloidal suspension of graphene oxide (GO) was used for obtaining the GO-mimic miRNA-7 nanosystems by self-assembly method. The ultrastructure, size distribution and ATR-FTIR and UV-Vis spectrum were analyzed. The Zeta potential was measured to verify the stability of obtained nanosystem. The entrapment efficiency, loading capacity and released kinetics of mimic miRNA-7 form GO-mimic miRNA-7 nanosystems were analyzed. The transfection efficiency into the glioblastoma cell lines U87, U118, U251, A172 and T98 of mimic miRNA-7 delivered by GO nanosystems was measure by confocal microscopy and flow cytometry. The changes at mRNA expression level of mTOR, PI3K, AKT1 and PTEN genes was measured by qPCR analysis. The xenograft model of U87 and A172 tumour tissue was performed to analyze the effect at tumor size and volume after GO- mimic miRNA-7 nanosystem administration.Results: The ultrastructure of GO-mimic miRNA-7 nanosystems showed high affinity of mimic miRNA into the GO. The results of transfection efficiency, cell morphology and viability showed that GO -miRNA-7 effectively deliver mimics miRNA-7 into U87, U118, U251, A172 and T98 glioblastoma cells. This approach can reverse miRNA-7 expression’s downstream effects and target the mTOR PI3K/Akt pathway observed at gene expression level, reducing xenograft tumour size and volume.Conclusion: The findings of the study could have significant implications for the development of advanced and precise GO based nanosystems specifically designed for miRNA therapy in cancer treatment.Keywords: GO, micro-RNA, cancer, mTOR/PI3K/Akt

Marlena Zielińska-Górska,1 Malwina Sosnowska-Ławnicka,1 Sławomir Jaworski,1 Agata Lange,1 Karolina Daniluk,1 Barbara Nasiłowska,2 Bartosz Bartosewicz,2 André Chwalibog,3 Ewa Sawosz1 1Department of Nanobiotechnology, Institute of Biology, Warsaw University of Life Sciences, Warsaw, 02-787, Poland; 2Institute of Optoelectronics, Military University of Technology, Warsaw, 00-908, Poland; 3Department of Veterinary and Animal Sciences, University of Copenhagen, Frederiksberg, 1870, DenmarkCorrespondence: Marlena Zielińska-Górska, Department of Nanobiotechnology, Institute of Biology, Warsaw University of Life Sciences, Ciszewskiego 8 St., bldg. 23, room 110, Warsaw, 02-786, Poland, Tel +48225936665, Email marlena_zielinska_gorska@sggw.edu.plBackground: Bacterial skin infections, including Staphylococcus aureus, are a powerful and still not fully resolved problem. The aim of this research was to determine the possibility of using a complex of graphene oxide (GO) encrusted with silver nanoparticles as an effective antibacterial agent against S. aureus and to assess its pro-inflammatory properties.Methods: The tests were carried out in vitro on EpiDerm™ Skin, an artificial skin model (MatTek in vitro Life Science Laboratories, Slovak Republic), and the fibroblast cell line (HFF-2 from ATCC, USA). Both models were infected with S. aureus bacteria (ATCC 25923) and then treated with antibiotics or our experimental factors: silver nanoparticles (AgNPs, Nano-koloid, Poland), graphene oxide (GO, NanoPoz, Poland), and complex AgNP-GO (hydrocolloid created by self-assembly).Results: The antibacterial effectiveness of the AgNP-GO complex was equivalent to that of the antibiotic. In addition, an increase in the level of pro-inflammatory cytokines was observed under the influence of antibiotic administration, in contrast to the effect of AgNP-GO, which showed very limited pro-inflammatory activity.Conclusion: Hydrocolloid of the AgNP-GO complex, administered in the form of a liquid dressing, may act as an antibacterial agent and also reduce inflammation induced by S. aureus infection.Keywords: human skin, inflammation state, nanomaterials, antibacterial properties

Purpose: Biofilms are one of the main threats related to bacteria. Owing to their complex structure, in which bacteria are embedded in the extracellular matrix, they are extremely challenging to eradicate, especially since they can inhabit both biotic and abiotic surfaces. This study aimed to create an effective antibiofilm nanofilm based on graphene oxide-metal nanoparticles (GOM-NPs)., Methods: To create nanofilms, physicochemical analysis was performed, including zeta potential (Zp) (and the nanocomposites stability in time) and size distribution measurements, scanning transmission electron microscopy (STEM), energy dispersive X-ray analysis (EDX), and atomic force microscopy (AFM) of the nanofilm surfaces. During biological analysis, reactive oxygen species (ROS) and antioxidant capacity were measured in planktonic cells treated with the nanocomposites. Thereafter, biofilm formation was checked via crystal violet staining, biofilm thickness was assessed by confocal microscopy using double fluorescent staining, and biofilm structure was analyzed by scanning electron microscopy., Results: The results showed that two of the three nanocomposites were effective in reducing biofilm formation (GOAg and GOZnO), although the nanofilms were characterized by the roughest surface, indicating that high surface roughness is unfavorable for biofilm formation by the tested bacterial species ( Staphylococcus aureus (ATCC 25923), Salmonella enterica (ATCC 13076), Pseudomonas aeruginosa (ATCC 27853))., Conclusion: The performed analysis indicated that graphene oxide may be a platform for metal nanoparticles that enhances their properties (eg colloidal stability, which is maintained over time). Nanocomposites based on graphene oxide with silver nanoparticles and other types of nanocomposites with zinc oxide were effective against biofilms, contributing to changes throughout the biofilm structure, causing a significant reduction in the thickness of the structure, and affecting cell distribution. A nanocomposite consisting of graphene oxide with copper nanoparticles inhibited the biofilm, but to a lesser extent., Competing Interests: The author(s) report no conflicts of interest in this work., (© 2024 Lange et al.)

Introduction and Objective: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) enters the nasal cavity, penetrates the nasal epithelial cells through the interaction of its spike protein with the host cell receptor angiotensin-converting enzyme 2 (ACE2) and then triggers a cytokine storm. We aimed to assess the biocompatibility of fullerenol nanoparticles C 60 (OH) 40 and ectoine, and to document their effect on the protection of primary human nasal epithelial cells (HNEpCs) against the effects of interaction with the fragment of virus - spike protein. This preliminary research is the first step towards the construction of a intranasal medical device with a protective, mechanical function against SARS-CoV-2 similar to that of personal protective equipment (eg masks)., Methods: We used HNEpCs and the full-length spike protein from SARS-CoV-2 to mimic the first stage of virus infection. We assessed cell viability with the XTT assay and a spectrophotometer. May-Grünwald Giemsa and periodic acid-Schiff staining served to evaluate HNEpC morphology. We assessed reactive oxygen species (ROS) production by using 2',7'-dichlorofluorescin diacetate and commercial kit. Finally, we employed reverse transcription polymerase chain reaction, Western blotting and confocal microscopy to determine the expression of angiotensin-converting enzyme 2 (ACE2) and inflammatory cytokines., Results: There was normal morphology and unchanged viability of HNEpCs after incubation with 10 mg/L C 60 (OH) 40 , 0.2% ectoine or their composite for 24 h. The spike protein exerted cytotoxicity via ROS production. Preincubation with the composite protected HNEpCs against the interaction between the spike protein and the host membrane and prevented the production of key cytokines characteristic of severe coronavirus disease 2019, including interleukin 6 and 8, monocyte chemotactic protein 1 and 2, tissue inhibitor of metalloproteinases 2 and macrophage colony-stimulating factor., Conclusion: In the future, the combination of fullerenol and ectoine may be used to prevent viral infections as an intranasal medical device for people with reduced immunity and damaged mucous membrane., Competing Interests: Dr Malwina Sosnowska, Prof Mateusz Wierzbicki, Dr Barbara Strojny-Cieślak, Prof Ewa Sawosz and Dr Marta Kutwin report a pending patent “Composition with barrier and virucidal properties against coronaviruses, especially SARS-CoV-2, method of obtaining it and use” (Poland patent pending, number P.448505). The authors declare no conflict of interest in this work., (© 2024 Sosnowska et al.)

This paper presents the results of studies on the effects of four types of aerosols containing an aqueous dispersed suspension of graphene oxide (GO) and an aqueous dispersed suspension of graphene oxide with the addition of curcumin (GO + C), silver nanoparticles (GO + Ag), and hypochlorous acid (GO + HClO) on selected structural and biological properties. Structural studies were carried out using electron microscopy, including a scanning electron microscope (SEM), scanning transmission electron microscopy (STEM), laser emission spectroscopy (LIBS), and absorption spectra in the infrared range attuned total reflectance (FTIR-ATR). The growth inhibition zone and viability of Staphylococcus aureus and Pseudomonas aeruginosa bacteria were studied. Studies have shown that the addition of silver nanoparticles and hypochlorous acid to the nanostructures of graphene oxide suspension improves bactericidal properties. In addition, it was observed that the application of a dispersed graphene oxide suspension in the form of an aerosol enriched with hypochlorous acid and silver nanoparticles results in the formation of a fairly uniform layer of graphene flakes, characterized by the presence of admixtures used.

In this work, pine and birch wood were modified by graphene oxide using a single vacuum impregnation method. The research results indicate that the impregnation of wood with graphene oxide increases the crystallinity of cellulose in both pine and birch wood, and the increase in crystallinity observed in the case of birch was more significant than in the case of pine. FT-IR analyses of pine samples impregnated with graphene oxide showed changes in intensity in the absorption bands of 400-600, 700-1500 cm -1 , and 3200-3500 cm -1 and a peak separation of 1102 cm -1 , which may indicate new C-O-C connections. In the case of birch, only some differences were noticed related to the vibrations of the OH group. The proposed modification also affects changes in the color of the wood surface, with earlywood containing more graphene oxide than latewood. Analysis of scanning electron microscope images revealed that graphene oxide adheres flat to the cell wall. Considering the differences in the anatomical structure of both wood species, the research showed a statistically significant difference in water absorption and retention of graphene oxide in wood cells. Graphene oxide does not block the flow of water in the wood, as evidenced by the absorbability of the working liquid at the level of 580-602 kg/m 3 , which corresponds to the value of pure water absorption by wood in the impregnation method using a single negative pressure. In this case, higher graphene oxide retention values were obtained for pine wood.

This research studied the phenolic content compared with the antioxidant properties of various O. vulgare (Lamiaceae) cultivars grown in Poland. The research results in this paper indicate that the dominant ingredient in all oregano cultivars was rosmarinic acid, known for its strong antioxidant properties. The highest amounts of rosmarinic acid (87.16 ± 4.03 mg/g dm) were identified in the O. vulgare spp. hirtum (Link) Ietsw. Other metabolites identified in the studied extracts include luteolin O-di-glucuronide-O-di-pentoside (30.79 ± 0.38 mg/g dm in the 'Aureum' cultivar), 4'-O-glucopyranosyl-3', 4'-dihydroxy benzyl-protocatechuate (19.84 ± 0.60 mg/g dm in the 'Margerita' cultivar), and p -coumaroyl-triacetyl-hexoside (25.44 ± 0.18 mg/g dm in the 'Margerita' cultivar). 'Hot & spicy' and 'Margerita' cultivars were characterized by the highest activity in eliminating OH • and O 2 •- radicals. Extracts from Greek oregano had the highest ability to scavenge DPPH radicals and chelate iron ions. This research has also provided new evidence that oregano has anti-migratory, cytotoxic properties and influences the viability of gastric cancer cells (the highest cytotoxicity was attributed to the 'Hot & spicy' cultivar, which performed the worst in antioxidant properties tests). Extracts from the tested cultivars at a concentration of 0.625% effectively inhibited the growth of S. aureus and P. aeruginosa bacteria. It seems that the oregano grown in Poland is of good quality and can be successfully grown on a large scale if the appropriate use is found.

This publication presents the effect of hypochlorous acid dry mist as a disinfectant on selected bacteria, viruses, spores, and fungi as well as on portable Microlife OXY 300 finger pulse oximeters and electronic systems of Raspberry Pi Zero microcomputers. The impact of hypochlorous acid on microbiological agents was assessed at concentrations of 300, 500, and 2000 ppm of HClO according to PN-EN 17272 (Variant I). Studies of the impact of hypochlorous acid fog on electronic components were carried out in an aerosol chamber at concentrations of 500 ppm and 2000 ppm according to two models consisting of 30 (Variant II) and 90 fogging cycles (Variant III). Each cycle included the process of generating a dry mist of hypochlorous acid (25 mL/m 3 ), decontamination of the test elements, as well as cleaning the chamber of the disinfectant agent. The exposure of the materials examined on hypochlorous acid dry mist in all variants resulted in a decrease in the number of viruses, bacteria, spores, and fungi tested. In addition, the research showed that in the variants of hypochlorous acid fogging cycles analyzed, no changes in performance parameters and no penetration of dry fog of hypochlorous acid into the interior of the tested medical devices and electronic systems were observed.

This paper presents the results of research on the impact of graphene paper on selected bacterial strains. Graphene oxide, from which graphene paper is made, has mainly bacteriostatic properties. Therefore, the main goal of this research was to determine the possibility of using graphene paper as a carrier of a medicinal substance. Studies of the degree of bacterial inhibition were performed on Staphylococcus aureus and Pseudomonas aeruginosa strains. Graphene paper was analyzed not only in the state of delivery but also after the incorporation of the antibiotics ciprofloxacin, cefazolin, and methicillin into its structures. In addition, Fourier-Transform Infrared Spectroscopy, contact angle, and microscopic analysis of bacteria on the surface of the examined graphene paper samples were also performed. Studies have shown that graphene paper with built-in ciprofloxacin had a bactericidal effect on the strains of Staphylococcus aureus and Pseudomonas aeruginosa . In contrast, methicillin, as well as cefazolin, deposited on graphene paper acted mainly locally. Studies have shown that graphene paper can be used as a carrier of selected medicinal substances.

The study aimed to test the durability of coatings under natural conditions. The present study focused on the changes in wettability and additional properties of the coatings under natural conditions. The specimens were subjected to outdoor exposure and additionally immersed in the pond. Impregnating porous anodized aluminum is a popular production method for hydrophobic and superhydrophobic surfaces. However, prolonged exposure of such coatings to natural conditions causes leaching of the impregnate and, thus, the loss of hydrophobic properties. After the loss of hydrophobic properties, all kinds of impurities and fouling adhere better to the porous structure. Additionally, deterioration of anti-icing and anti-corrosion properties was observed. Finally, the self-cleaning, anti-fouling, anti-icing and anti-corrosion properties were comparable or even worse to those of the hydrophilic coating. In the case of superhydrophobic specimens, during outdoor exposure there was no loss of superhydrophobicity, self-cleaning and anti-corrosion properties. Still, despite this, the icing delay time dropped. During outdoor exposure, the structure, which initially had anti-icing properties, may degrade. Nevertheless, the hierarchical structure responsible for the superhydrophobic effect can still be preserved. The superhydrophobic coating initially had the best anti-fouling properties. However, the coating was also gradually losing its superhydrophobic properties during water immersion.

This paper presents the results of an analysis of carbon (in the form of graphene oxide) deposited on the surface of threads made from stainless steel 316 and titanium alloy Ti6Al4V used in orthopedics using Laser Induced Breakdown Spectroscopy (LIBS). The aim of the article is to indicate the possibility of using the LIBS spectra for the study of thin layers, including graphene derivatives and other elements. Stratigraphic measurements allowed the detection of differences in the spectra peaks of individual elements, not only in the surface layer itself and in the native material, but also in the intermediate layer connecting the two layers. Due to the clear difference in the outline of the spectrum of graphene oxide and the spectrum of the native material of the samples analyzed, a clear incorporation of carbon atoms into the surface layer was observed. A factor analysis was performed, which confirmed the incorporation of graphene oxide into the surface layer of the native material of the elements examined.

A material strength investigation along with a detailed microfractography analysis of fractures formed during static tensile tests of steel Armstal 550 was performed. The tests in this research were conducted in a temperature range of 298 to 973 K. In addition, during tensile tests at ambient temperature, optical measurements of strain maps and the curvature of the neck were performed. The minimum cross-sectional diameter and the radius of the neck curvature during tensile tests were obtained. The data can be directly used to obtain the true stress-strain curve. The material property analysis confirmed the high strength of the Armstal 550 alloy. The ultimate strength at room temperature equals 2.14 GPa, whereas the yield point equals 1.65 GPa. A decrease in the strength parameters along with an increase in temperature was noted. This is a typical phenomenon related to a change in the density and thermal expansion of steel under the influence of the temperature increase. For example, at a temperature of 500 °C, the ultimate strength is more than 50% less than at room temperature. An in-depth analysis of the metallography and microfractography of fractures resulting from static tensile tests showed the formation of atypical nano- and microstructures with an elongated shape. Local nano- and microstructures were observed at different levels of intensity for different temperatures. The largest clusters of nanoparticles were present on the surfaces of the specimens examined at a temperature of 973 K. Scanning microscopy analysis confirmed the presence of molybdenum oxides.

This work shows the effect of graphene oxide deposition on microsieves' surfaces of gold and nickel foils, on DU 145 tumor cells of the prostate gland. The sieves were made by a laser ablation process. The graphene oxide (GO) deposition process was characterized by the complete covering of the inner edges of the microholes and the flat surface between the holes with GO. Electron microscanning studies have shown that due to the deposition method applied, graphene oxide flakes line the interior of the microholes, reducing the unevenness of the downstream surfaces during the laser ablation process. The presence of graphene oxide was confirmed by Fourier infrared spectroscopy. During the screening (sieving) process, the microsieves were placed in a sieve column. Gold foil is proven to be a very good material for the screening of cancer cells, but even more so after screening as a substrate for re-culture of the DU 145. This allows a potential recovery of the cells and the development of a targeted therapy. The sieved cells were successfully grown on the microsieves used in the experiment. Graphene oxide remaining on the surface of the nickel sieve has been observed to increase the sieving effect. Although graphene oxide improved separation efficiency by 9.7%, the nickel substrate is not suitable for re-culturing of the Du 145 cells and the development of a targeted therapy compared to the gold one.

The aim of this study was to obtain a superhydrophobic coating by modifying anodized aluminum using polydimethylsiloxane (PDMS). In order to obtain a superhydrophobic coating on an aluminum substrate, a multistage treatment was implemented. Specimens of aluminum were treated by abrasive blasting, anodization in sulfuric acid, impregnation by PDMS, rinsing in toluene to remove excess of PDMS, and curing. A rough surface with an additional low free energy layer on it resulted in a superhydrophobic effect. The coating obtained has an average contact angle of 159°. The specimens were tested in terms of durability in natural conditions. Additionally, anti-icing and anti-fouling properties were evaluated. The coating was compared with anodized aluminum obtained by a basic process.

This paper presents the effect of deposited graphene oxide coating on fatigue life of austenitic steel 1.4541 at 20 °C, 100 °C, and 200 °C. The study showed a decrease in the fatigue life of samples with a deposited graphene oxide layer in comparison with reference samples at 20 °C and 100 °C. However, an increase in fatigue life of samples with a deposited graphene oxide layer in comparison with reference samples occurred at 200 °C. This relationship was observed for the nominal stress amplitude of 370 and 420 MPa. Measurements of temperature during the tensile failure of the sample and microfractographic analysis of fatigue fractures were performed. Tests have shown that graphene oxide deposited on the steel surface provides an insulating layer. A higher temperature of the samples with a deposited graphene oxide layer was observed during fracture compared to the reference samples.

A near 1-keV photons from the Xe/He plasma produced by the interaction of laser beam with a double stream gas puff target were employed for studies of L absorption edges of period 4 transitional metals with atomic number Z from 26 to 30. The dual-channel, compact NEXAFS system was employed for the acquisition of the absorption spectra. L 1-3 absorption edges of the samples were identified in transmission mode using broadband emission from the Xe/He plasma to show the applicability of such source and measurement system to the NEXAFS studies of the transition metals, including magnetic materials.

This paper presents the results of investigations of the effect of graphene oxide and surface shot peening on the mechanical properties and fatigue life of bolts made of austenitic 304 steel. An innovative method for the uniform deposition of graphene oxide on screws is presented. The process involved activating the surface using plasma and then performing graphene oxide deposition using centrifugal force and vacuum drying. The screw specimens prepared in this way were subjected to a surface peening process. Comparative studies have shown that the combination of graphene oxide deposition and shot peening processes results in an increase in fatigue life of approximately 42 ÷ 275% (depending on the stress amplitude level) compared to the as-delivered samples. The results presented are promising and may provide a basis for further research on the application of graphene and its derivatives to increase fatigue life and improve the mechanical properties of machine components.

This paper presents the results of the interaction of graphene oxide (GO) on MDA-MB-231 and SW-954 cancer cell lines. The tests were carried out in two variants. In the first one, GO was sprayed on a Petri dish and then, the cancer cell lines were cultured. In the second variant, the cells were covered with an aerosol containing GO. In both variants, cancer cell lines were incubated and tested every 24, 48, and 72 h. After each time period, cell viability and surface morphology were measured. The tests after 72 h showed that coating with GO aerosol caused a reduction in cell viability by 52.7% and 26.4% for MDA-MB-231 and SW-954 cancer cell lines, respectively, with respect to a reference sample (without the influence of GO aerosol). Tests where GO is a culture medium demonstrated a decrease in cell viability by approximately 4.3% compared to a reference sample for both considered cell lines.

The graphene paper microsieves can be applied in the filtration of biological fluids or separation of solid particles from exploitation fluids. To produce graphene paper microsieves for specific applications, good control over fabrication should be achieved. In this study, a laser ablation method using a picosecond laser was applied to fabricate graphene paper microsieves. Holes in the microsieves were drilled using pulsed laser radiation with a pulse energy from 5 to 100 µJ, a duration of 60 ps, a wavelength of 355 nm, and a repetition rate of 1 kHz. The impact method was applied using 10 to 100 pulses to drill one hole. To produce holes of a proper diameter which could separate biological particles of a certain size (≥10 µm), optimum parameters of graphene paper laser ablation were defined using the MATLAB software taking into account laser pulse energy, repetition rate, and a desired hole diameter. A series of structural tests were carried out to determine the quality of an edge and a hole shape. Experimental results and Laguerre-Gauss calculations in MATLAB were then compared to perform the analysis of the distribution of diffraction fringes. Optimum experimental parameters were determined for which good susceptibility of the graphene paper to laser processing was observed., Competing Interests: The authors declare no conflict of interest.

Background: The accessibility of the remineralizing ions in teeth's environment is essential for their incorporation into caries-affected dentin. Novel bioglass-reinforced materials capable of releasing fluoride, calcium and phosphates may be particularly useful in the tissue remineralization process. A novel restorative material, ACTIVA BioActive-Restorative (Pulpdent Corp., Watertown, USA), is a hydrophilic resin-modified glassionomer cement (RMGIC) enriched with bioglass particles and fortified with a patented rubberized polymer resin. Its application in restorative dentistry may be significant, promoting remineralization of carious lesions., Objectives: The aim of the study was to compare the fluoride ion release profiles from a bioglass-reinforced RMGIC, a conventional glass-ionomer cement (GIC) and a nanohybrid restorative polymer resin., Material and Methods: The quantity of fluoride ions released from ACTIVA, Ketac Molar Quick Aplicap and Tetric EvoCeram was assessed using a fluoride-specific electrode. The surface characteristics of the preand post-experimental specimens were studied using a scanning electron microscope (SEM) and confocal microscope. An X-ray powder diffraction (XRD) analysis was additionally used to examine the chemical compositions of the dental materials., Results: The greatest quantity of fluoride ions was freed from the GIC specimens (20.698-54.118 ppm), followed by the bioglass-reinforced RMGIC (from 1.236 to 15.552 ppm) and nanohybrid polymer resin (0.370-1.148 ppm). The pre-experimental specimens of the bioglass-reinforced RMGIC were porous, while the post-experimental specimens were smoother with visible micro-cracks. The XRD analysis of the bioglass particles confirmed that the material was composed mainly of fluoride (27.70 mass%), silicon (15.62 mass%), aluminum (5.91 mass%), and calcium (5.40 mass%)., Conclusions: The fluoride ion release profile of ACTIVA was lower than the GIC Keta Molar Quick Aplicap, but significantly higher than the nanohybrid restorative polymer resin Tetric EvoCeram.

The results of studies on the fabrication and characterization of silver nanoisland films (SNIFs) using pulsed laser deposition (PLD) and the evaluation of these films as potential surface-enhanced Raman scattering (SERS) substrates are reported. The SNIFs with thicknesses in a range of 4.7 ± 0.2 nm to 143.2 ± 0.2 nm were deposited under different conditions on silicon substrates. Size and morphology of the fabricated silver nanoislands mainly depend on the substrate temperature, and number and energy of the laser pulses. SERS properties of the fabricated films were evaluated by measuring SERS spectra of para -mercaptoaniline (pMA) molecules adsorbed on them. SERS enhancement factors are shown to depend on the SNIF morphology, which is modified by changes of the deposition conditions. The highest enhancement factor in the range of 10 5 was achieved for SNIFs that have oval and circular silver nanoislands with small distances between them.

Background: Graft-versus-host-disease (GvHD) is the major cause of morbidity and mortality after stem cell transplantation. The development of early prediction methods is therefore of importance. Our aim was to analyze the usefulness of early donor chimerism monitoring after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in T cells and in CD4+ and CD8+ (lineage chimerism) for GvHD prediction., Material and Methods: Chimerism was analyzed in 76 consecutive adult patients using RQ-PCR TaqMan technology on DNA extracted from Pan T, CD4+, and CD8+ cell subsets on Day 5, 10, 15 and 30 after allo-HSCT., Results: The threshold of chimerism predictive for GvHD was the same for all tested cell subsets. In acute myeloid leukemia (AML) patients treated with myeloablative conditioning (MAC), the threshold predictive for acute graft versus host disease was 95% and 99% for Day 10 and Day 15, respectively. In patients treated with reduced intensity conditioning (RIC), the threshold predictive for chronic graft versus host disease was 98% on Day 10. The differences were statistically significant., Conclusions: Chimerism analysis in T cell subsets by RQ-PCR on Day 10 and Day 15 after transplantation is useful for prediction of aGvHD (AML patients after MAC) and cGvHD (patients after RIC). However, there was no difference in the results between chimerism in the T cell subsets. Our RQ-PCR protocol was highly sensitive and proved effective for analysis of lineage chimerism.

Unlabelled: It is recommended that all patients after allogeneic hematopoietic stem cell transplantation (alloHSCT) should be monitored for CMV infection markers. The aim of the study was to check the usefulness of quantitative DNA CMV monitoring after alloHSCT., Material and Method: DNA CMV was tested by real-time PCR in sera and blood samples twice a week until 30th day after alloHSCT thereafter, once a week until 100th day and then, once every 2-3 weeks. 832 samples from 16 patients were tested. All patients were anti-CMV positive or/and received stem cells from seropositive donors. Introduction of antiviral treatment was based on initial viral load and its rate of increase., Results: DNA CMV was detected in 13/16 patients; in 3 before 30h day after allo HSCT (group I) and in 10 (group II) after 30th day. In all patients from group I clinical symptoms were observed and DNA CMV was detected in sera and blood samples. Peak viral load was 2490-34 620 geq/ml. Although antiviral treatment was applied, reinfection was observed and infection lasted from 28 to 91 days. In 6 group II patients, clinical symptoms were observed and DNA CMV in sera and blood was detected for 16-56 days, DNA CMV peak load was 100-8950 geq/ml. In the remaining 4 patients, no clinical symptoms were observed--DNA CMV was detected in blood only for 7 to 14 days. In one patient with peak viral load 10,540 geq/ml, antiviral treatment was applied. In 3 with viral load of 400-2000 geq/ml, treatment was not introduced. The quantitative DNA CMV results were taken into account before the change of antiviral drugs for more effective drugs and the decrease of drug dose due to side effects., Conclusions: Application of quantititative DNA CMV testing allowed to optimise antiviral drug administration in immunosupressed patients after alloHSCT

Background: Osteolytic bone destruction caused by increase of osteolytic activity is a major manifestation of multiple myeloma (MM). Pamidronate (3-amino-1-hydroxypropylidene)-1,1-bisphosphonate) inhibits osteoclastic activity and reduces bone resorption., Methods: Since October 1995 the efficacy of pamidronate is evaluated in MM patients all receiving anti-myeloma chemotherapy acc. to VMCP/VBAP alternating regimen. 46 patients with stage III myeloma and osteolytic lesions were randomized to receive either pamidronate (Aredia; Novartis) 60 mg i.v. in 4-hour infusion monthly (n = 23) or chemotherapy alone (control group n = 23). Estimation of performance status, quality of life, pain score, analgesic consumption, serum calcium concentration and twenty four-hours Calcium excretion, urine Calcium/creatinine ratio is done at least once a month (before pamidronate administration) while X-ray skeletal survey--before treatment and then every six months., Results: In the first months of treatment apparent reduction of bone pain occurred. Hypercalcaemia was revealed in 6 patients at entry into the study. In 5 of these patients pamidronate restored and maintained normocalcaemia for a median 6 months. In 3 patients an aggressive plasma cell proliferation was accompanied by reoccurrence of hypercalcaemia. At skeletal X-ray examination performed after 6 and 12 cycles of pamidronate and by comparing each of consecutive imaging with previous one the progression of osteolysis was respectively found in 67% and 39% of patients. In the control group corresponding figures were: 79% and 70%. The mean number of skeletal events (pathologic fracture, radiation to bone and spinal cord compression) per year was lower in the pamidronate group (1.82) than in control-patients (2.72), p < 0.013. The proportion of patients who developed skeletal event (excluding vertebral fractures) was lower in the pamidronate group -34% v 52%. Adverse events of pamidronate: hypocalcaemia (< 2 mmol/l) observed in 7 patients occurred in particular patients beginning from 2 to 7 days after drug administration. In 2 patients hypocalcaemia that appeared in 24 hours after drug infusion was accompanied by blood pressure decrease; in one case systolic blood pressure dropped up to 60 mmHg, in the other one--to 90 mmHg. Muscular pain and fever up to 39 degrees C (transient and self-limiting "influenza like syndrom") occurred in 5 patients, in two patients after several hours and in three other--after some dozens of hours from drug administration. In one case hypertransaminasaemia was observed., Conclusions: In the first year of treatment monthly intravenous pamidronate administration as an adjunct to chemotherapy in patients with advanced multiple myeloma with osteolysis is an efficient approach in prevention and treatment of hyperacalcaemia, hypercalciuria and bone pain. It also shows some preventive effect on bone lesion occurrence.