The Impact of the Emotional Disposition of Healthcare Workers on the Expression of Adverse Events after Primary Vaccination against SARS-CoV-2.

Background and Objectives: Vaccination is one means of SARS-CoV-2 prevention and control. However, despite the effectiveness of vaccination, adverse reactions continue to require vigilance and monitoring. The researchers emphasize the possibility that some of the reported side effects may be psychological in origin. Based on this hypothesis, the main goal of this study was to evaluate the emotional dispositions of healthcare workers who experienced emotions before vaccination and adverse reactions after vaccination. Materials and Methods: This study was conducted between February and May 2021 in the Kaunas Clinics of the University of Health Sciences. A total of 2117 employees of the clinic departments who were vaccinated with two doses of the Pfizer-BioNTech vaccine participated in this study. Statistical analysis was performed on the data using IBM SPSS Statistics®. Results: Most participants (74.5%) experienced systemic (including local) adverse events; 16.5% experienced only local adverse events, and 9.1% experienced no adverse events. The frequency of systemic (including local) adverse events reduced with increasing age (p < 0.05). The main emotions that participants experienced before vaccination were anxiety (37.88%) and happiness (39.02%). Systemic (including local) adverse events occurred 1.26 times more frequently in women than men (77.44% vs. 61.6%, p < 0.05), while local adverse events occurred 1.4 times more often in male participants than in female participants (21.39% vs. 15.27%, p < 0.05). Among the respondents who did not experience adverse events, the most common emotion felt was happiness (25.5%), and most of the participants who experienced systemic (including local) adverse events felt anxiety (42.6%). Conclusions: The information about vaccination and potential adverse events should be targeted at younger persons. It is recommended that women, more than men, should receive professional counseling from psychologists or psychotherapists. The public dissemination of positive messages about the benefits and safety of vaccines prior to a vaccination campaign may alleviate the tension or anxiety felt regarding potential adverse events. Healthcare specialists-both those who work directly with vaccines and those who do not-should maintain a positive psychological attitude towards vaccination, as this can increase patient satisfaction with the benefits of vaccines.

Ethidium Binding to Salmonella enterica ser. Typhimurium Cells and Salmon Sperm DNA.

Bacterial resistance to antibiotics due to increased efficiency of the efflux is a serious problem in clinics of infectious diseases. Knowledge of the factors affecting the activity of efflux pumps would help to find the solution. For this, fast and trustful methods for efflux analysis are needed. Here, we analyzed how the assay conditions affect the accumulation of efflux indicators ethidium (Et+) and tetraphenylphosphonium in Salmonella enterica ser. Typhimurium cells. An inhibitor phenylalanyl-arginyl-ß-naphtylamide was applied to evaluate the input of RND family pumps into the total efflux. In parallel to spectrofluorimetric analysis, we used an electrochemical assessment of Et+ concentration. The results of our experiments indicated that Et+ fluorescence increases immediately after the penetration of this indicator into the cells. However, when cells bind a high amount of Et+, the intensity of the fluorescence reaches the saturation level and stops reacting to the accumulated amount of this indicator. For this reason, electrochemical measurements provide more trustful information about the efficiency of efflux when cells accumulate high amounts of Et+. Measurements of Et+ interaction with the purified DNA demonstrated that the affinity of this lipophilic cation to DNA depends on the medium composition. The capacity of DNA to bind Et+ considerably decreases in the presence of Mg2+, Polymyxin B or when DNA is incubated in high ionic strength media.

Interaction of ethidium and tetraphenylphosphonium cations with Salmonella enterica cells.

BACKGROUND AND OBJECTIVE: One of the main causes of bacterial resistance to antimicrobials is multidrug resistance induced by the increased efficiency of the efflux pumps. In this study we analyzed how the conditions of assay affect the efflux of indicator substrates ethidium (Et+) and tetraphenylphosphonium (TPP+) in Salmonella enterica ser. Typhimurium cells. Impact of the outer membrane permeability barrier, composition and temperature of the medium on accumulation of the indicator compounds also was analyzed. MATERIALS AND METHODS: The fluorescence of Et+ and Nile Red was measured using 96-well plates and a plate reader. In parallel to traditional studies of fluorescence we applied a constructed selective electrode to follow the accumulation of Et+ in S. enterica cells. Simultaneously with monitoring of Et+ concentration in the cell incubation medium, electrochemical measurements of TPP+ accumulation were performed. Furthermore, Et+ and TPP+ were used within the same sample as agents competing for the interaction with the efflux pumps. An inhibitor phenylalanyl-arginyl-ß-naphtylamide (PAßN) was applied to evaluate the input of RND-family pumps in the total efflux of these indicator compounds. RESULTS: S. enterica cells with the intact outer membrane (OM) bound very low amounts of Et+ or TPP+. Cells with the permeabilized OM accumulate considerably higher amounts of the indicator compounds at pH 8.0, but only Et+ was considerably accumulated at pH 6.5. At conditions of electrochemical monitoring accumulation of Et+ by the permeabilized cells at 37°C was considerably faster than at 23°C, but at the higher temperature most of the cell-accumulated Et+ was extruded back to the medium. The fluorescence of Et+ in suspension of cells incubated in 400mmol/L Tris buffer was about twice higher compared to 100mmol/L one. The inhibitory action of TPP+ on Et+ efflux was evident only in 400mmol/L Tris although PAßN effectively increased Et+ fluorescence at both buffer concentrations. CONCLUSIONS: Results of our experiments indicate that ionic strength of the incubation medium influence the selectivity, the medium temperature and the assay conditions impact the kinetics of efflux. The lower accumulated amount and the weaker fluorescence of Et+ registered in slightly acidic medium indicate that ΔΨ plays a role in the accumulation of this indicator cation. The bound amount of Et+ to the de-energized or permeabilized cells considerably varies depending on the conditions and methods of de-energization or permeabilization of cells. Tris/EDTA permeabilization of the cells does not inhibit the efflux.

Mode of the Interaction of Efflux Inhibitor Phenylalanyl-arginyl-ß-naphtylamide with Bacterial Cells.

An increased efflux activity is one of the major reasons for bacterial antibiotic resistance. The usage of efflux pump inhibitors could be a promising approach to restoring the activity of inefficient antibiotics. The interaction of the RND family efflux pump inhibitor phenylalanyl-arginyl-ß-naphthylamide (PAßN) with Salmonella enterica ser. Typhimurium cells was assayed using traditional microbiological techniques and a novel PAßN-selective electrode. Monitoring the PAßN concentration in the medium using the electrode enabled the real-time measurements of this compound's interaction with bacterial cells. We showed that S. Typhimurium cells accumulate a high amount of PAßN because of its high affinity to lipopolysaccharides (LPSs), the major constituent of the outer layer of the outer membrane, and does not affect the functioning of the plasma membrane. EDTA enhanced the binding of PAßN to S. Typhimurium cells and the purified E. coli LPSs, but the energization of the cells by glucose does not affect the cell-bound amount of this inhibitor. Polycationic antibiotic Polymyxin B released both the cells accumulated and the suspended LPS-bound PAßN.

Immune Response after Anti-SARS-CoV-2 mRNA Vaccination in Relation to Cellular Immunity, Vitamin D and Comorbidities in Hemodialysis Patients.

In the global threat of SARS-CoV-2, individuals undergoing maintenance dialysis represent a vulnerable population with an increased risk of severe COVID-19 outcomes. Therefore, immunization against SARS-CoV-2 is an essential component of healthcare strategy for these patients. Existing data indicate that they tend to exhibit a reduced immune response to vaccines compared to the general population. Our study aimed to assess both humoral and cellular immune responses following two doses of an anti-SARS-CoV-2 mRNA vaccine, an ability to maintain adequate antibody titers over time, and potential relations with vitamin D, comorbidities and other factors in hemodialysis patients based on a single center experience. A total of 41/45 patients (91.1%) responded to the second dose of the anti-SARS-CoV-2 mRNA vaccine. The titer of anti-SARS-CoV-2 IgG class antibodies and levels of T cells three to four weeks after vaccination were lower in dialysis patients than in healthy controls. Antibodies titer in dialysis patients had a positive correlation with B lymphocytes and was related to cardiovascular diseases. The level of CD4+ cells had a negative correlation with hemodialysis vintage, as did the vitamin D level with post-vaccination seroconversion and decline in anti-SARS-CoV-2 antibodies titer during six months after vaccination. Hemodialysis patients had decreased amounts of CD4+ and CD8+ cells and lower levels of anti-SARS-CoV-2 antibodies than healthy controls. Therefore, chronic hemodialysis could lead to diminished cellular immunity and humoral immune response to the anti-SARS-CoV-2 mRNA vaccination and reduced protection from COVID-19. Comorbidity in cardiovascular diseases was associated with a lower level of specific anti-SARS-CoV-2 antibody titer. Vitamin D may be important in maintaining stable levels of anti-SARS-CoV-2 antibodies, while the duration of dialysis treatment could be one of the factors decreasing anti-SARS-CoV-2 antibody titer and determining lower CD4+ cell counts.

Prevalence of COVID-19 in Kidney Transplant Patients in Relation to Their Immune Status after Repeated Anti-SARS-CoV-2 Vaccination.

The prospective study was conducted to evaluate the prevalence of COVID-19 in kidney transplant patients in relation to their immune status after three doses of the BNT162b2 (Pfizer-BioNTech) vaccine during one post-pandemic year based on the experience of one center-Hospital of Lithuanian University of Health Sciences. Thirty-three patients were invited for a follow-up visit 3 to 6 weeks after anti-SARS-CoV-2 vaccination and were obliged to report having COVID-19 during the one-year post-pandemic period. Forty-two percent of patients developed antibody response against SARS-CoV-2 after the third dose of the vaccine. The number of COVID-19 cases during the post-pandemic period did not differ significantly between seropositive and seronegative patients. However, only seronegative patients were hospitalized due to COVID-19. The anti-SARS-CoV-2 antibody titer in seropositive patients correlated with a relative number of CD3+ cells (R = 0.685, p = 0.029). The CD8+/CD38+ ratio in this group increased 2-fold after the anti-SARS-CoV-2 vaccination. Higher antibody response to the COVID-19 vaccine was associated with better kidney function. The anti-SARS-CoV-2 antibody titer relation with the components of cellular immunity (CD3+ cells and CD8+/CD38+ ratio) shows a role of both chains during the response to the anti-SARS-CoV-2 vaccine in kidney transplant patients.

Visible-Light-Driven Photocatalytic Inactivation of Bacteria, Bacteriophages, and Their Mixtures Using ZnO-Coated HDPE Beads as Floating Photocatalyst.

Semiconductor materials used as photocatalysts are considered among the most effective ways to treat biologically polluted water. Certainly, efficiency depends on the selection of photocatalyst and its substrate, as well as the possibility of its application in a broader spectrum of light. In this study, a reactive magnetron sputtering technique was applied for the immobilisation of ZnO photocatalyst on the surface of HDPE beads, which were selected as the buoyant substrates for enhanced photocatalytic performance and easier recovery from the treated water. Moreover, the study compared the effect on the inactivation of the microorganism between ZnO-coated HDPE beads without Ni and with Ni underlayer. Crystal structure, surface morphology, and chemical bonds of as-deposited ZnO films were investigated by X-ray diffraction, scanning electron microscopy, and X-ray photoelectron spectroscopy, respectively. Visible-light-induced photocatalytic treatment was performed on the Gram-negative and Gram-positive bacteria and bacteriophages PRD1, T4, and their mixture. Higher bacteria inactivation efficiency was obtained using the ZnO photocatalyst with Ni underlayer for the treatment of S. Typhimurium and M. Luteus mixtures. As for infectivity of bacteriophages, T4 alone and in the mixture with PRD1 were more affected by the produced photocatalyst, compared with PRD1.

Sustainable green strategy for recovery of glucose from end-of-life euro banknotes.

Usually, Euro banknotes are made from cotton substrates and their waste is disposed of in landfill or is incinerated. In order to valorize the end-of-life euro banknotes (ELEBs), the substrates were used in this research for cellulase production via submerged fungal fermentation (SFF), and the resultant fungal cellulase w s used in ELEBs hydrolysis process for extraction of glucose. The experiments were started by exposing the ELEBs to different types of pretreatments, including milling process, alkali (NaOH/urea solution), and acid leaching to remove any contamination (e.g. dyes) and to decrease the crystallinity of cellulose (the main element in cotton substrate) thus increasing the degradation rate during the fermentation process. The effect of pretreatments on the morphology and chemical composition of ELEBs was observed using Scanning Electron Microscope and Energy Dispersive Spectrometry. Afterwards, Trichoderma reesei-DSM76 was used for cellulase production from the treated ELEBs with high cellulase activity (12.97 FPU/g). The resultant cellulase was upscaled in a bioreactor and used in ELEBs hydrolysis. Finally, the results showed that the optimized pretreatment methods (milling followed by leaching process) significantly improved the cellulase activity and glucose recovery, which was estimated by 96%. According to the obtained results, the developed strategy has a great potential for conversion of ELEBs into a glucose product that could be used in biofuels and bioplastics applications.

Photocatalytic Inactivation of Salmonella typhimurium by Floating Carbon-Doped TiO2 Photocatalyst.

Photocatalysis application is considered as one of the most highly promising techniques for the reduction in wastewater pollution. However, the majority of highly efficient photocatalyst materials are obtained as fine powders, and this causes a lot of photocatalyst handling and reusability issues. The concept of the floating catalyst proposes the immobilization of a photocatalytic (nano)material on relatively large floating substrates and is considered as an encouraging way to overcome some of the most challenging photocatalysis issues. The purpose of this study is to examine floating photocatalyst application for Salmonella typhimurium bacteria inactivation in polluted water. More specifically, high-density polyethylene (HDPE) beads were used as a photocatalyst support for the immobilization of carbon-doped TiO2 films forming floating photocatalyst structures. Carbon-doped TiO2 films in both amorphous and anatase forms were deposited on HDPE beads using the low-temperature magnetron sputtering technique. Bacteria inactivation, together with cycling experiments, revealed promising results by decomposing more than 95% of Salmonella typhimurium bacteria in five consecutive treatment cycles. Additionally, a thorough analysis of the deposited carbon-doped TiO2 film was performed including morphology, elemental composition and mapping, structure, and depth profiling. The results demonstrate that the proposed method is a suitable technique for the formation of high-quality photocatalytic active films on thermal-sensitive substrates.

Antifungal activity of styrylpyridinium compounds against Candida albicans.

We synthesized a set of 13 new and earlier described styrylpyridinium compounds (N-alkyl styrylpyridinium salts with bromide or tosylate anions) in order to evaluate antifungal activity against C. albicans cells, to assay the possible synergism with fluconazole, and to estimate cytotoxicity to mammalian cells. All compounds were synthesized according to a well-known two-step procedure involving alkylation of γ-picoline with appropriate alkyl bromide and further condensation with substituted benzaldehyde. Compounds with long N-alkyl chains (C18 H37 -C20 H41 ) had no antifungal activity against the cells of all tested C. albicans strains. Other styrylpyridinium compounds were able to inhibit yeast growth at the concentrations of 0.06-16 µg/ml. At fungicidal concentrations, the compound with the CN- group was least toxic to mammalian cells, showed the most effective synergism with fluconazole, and only slightly inhibited the respiration of C. albicans. The compound with the 4'-diethylamino group exhibited the strongest fungicidal properties and effectively blocked the respiration of C. albicans cells. However, toxicity to mammalian cells was also high. Summarizing, the results of our study indicate that styrylpyridinium compounds are promising candidates in the development of new antifungal drugs.

Analysis of the Oxidative Stress Regulon Identifies soxS as a Genetic Target for Resistance Reversal in Multidrug-Resistant Klebsiella pneumoniae.

In bacteria, the defense system deployed to counter oxidative stress is orchestrated by three transcriptional factors, SoxS, SoxR, and OxyR. Although the regulon that these factors control is known in many bacteria, similar data are not available for Klebsiella pneumoniae. To address this data gap, oxidative stress was artificially induced in K. pneumoniae MGH78578 using paraquat and the corresponding oxidative stress regulon recorded using transcriptome sequencing (RNA-seq). The soxS gene was significantly induced during oxidative stress, and a knockout mutant was constructed to explore its functionality. The wild type and mutant were grown in the presence of paraquat and subjected to RNA-seq to elucidate the soxS regulon in K. pneumoniae MGH78578. Genes that are commonly regulated both in the oxidative stress and soxS regulons were identified and denoted as the oxidative SoxS regulon; these included a group of genes specifically regulated by SoxS. Efflux pump-encoding genes and global regulators were identified as part of this regulon. Consequently, the isogenic soxS mutant was found to exhibit a reduction in the minimum bactericidal concentration against tetracycline compared to that of the wild type. Impaired efflux activity, allowing tetracycline to be accumulated in the cytoplasm to bactericidal levels, was further evaluated using a tetraphenylphosphonium (TPP+) accumulation assay. The soxS mutant was also susceptible to tetracycline in vivo in a zebrafish embryo model. We conclude that the soxS gene could be considered a genetic target against which an inhibitor could be developed and used in combinatorial therapy to combat infections associated with multidrug-resistant K. pneumoniae. IMPORTANCE Antimicrobial resistance is a global health challenge. Few new antibiotics have been developed for use over the years, and preserving the efficacy of existing compounds is an important step to protect public health. This paper describes a study that examines the effects of exogenously induced oxidative stress on K. pneumoniae and uncovers a target that could be useful to harness as a strategy to mitigate resistance.

Floating TiO2 photocatalyst for efficient inactivation of E. coli and decomposition of methylene blue solution.

The anatase phase TiO2 films with nanocrystalline structure were successfully deposited on a water-floating non-expanded polystyrene (PS) beads via magnetron sputtering. The combination of UVB light and PS beads with TiO2 film was used for decomposition of methylene blue as well as inactivation tests for intact and EDTA-treated Escherichia coli bacteria. Crystal structure, elemental composition, elemental mapping, surface morphology and chemical bonds of TiO2 film were investigated. E. coli inactivation experiments showed that such floating photocatalyst could destroy >90% bacteria in 45 min under UVB irradiation. Results demonstrated that combination of TiO2 and UVB light leads to disruption of the outer membrane which causes effective inactivation of E. coli bacteria.

Antimicrobial copper nanoparticles synthesized from waste printed circuit boards using advanced chemical technology.

Waste Printed Circuit Boards (WPCBs) were classified as one of the most important resources for urban mining containing high purity Copper (Cu) and other valuable materials. Recently, a dissolution recycling approach enhanced by ultrasonic treatment succeeded in the liberation of Cu foils from WPCBs as received. This research aims to synthesize Copper Nanoparticles (Cu-NPs) from the recovered Cu by using an advanced chemistry approach to obtain nano-product with high added value taking into consideration environmental risks. The experiments were carried out on the Cu foils recovered from the three types of WPCBs with different purity of Cu (Motherboard, Video Card, and Random Access Memory (RAM)). The synthesis process was performed in two stages: (a) preparation of Copper (II) Sulfate aqueous solutions from the recovered Cu and (b) chemical reduction of solutions for synthesis of Cu-NPs by using Native Cyclodextrins (NCDs), particularly ß-NCD as stabilizers. The efficiency of the developed approach for raw material of different purity was assessed and the final yield and the estimated recovery cost of synthesized Cu-NPs were calculated with high accuracy as well as the properties of the synthesized Cu-NPs. The obtained Cu-NPs were examined using SEM-EDS, TEM, XRD, Raman Spectroscopy, and TGA. To maximize the potential biomedical application benefits, the antibacterial activity of Cu-NPs was investigated by the standard microdilution method for E. coli, P. aeruginosa, and S. aureus bacterial cultures. The results showed that the produced Cu-NPs had an average size of 7 nm and yield 90%, while the preparation costs were 6 times lower in comparison to the commercial counterparts. In addition, the results indicated that the synthesized Cu-NPs from RAM sample had a good antimicrobial action.