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OBJECTIVES The aim of the study was to analyze some metalloproteinases, cytokines, and chemokines in LB patients and healthy seropositive subjects. The presence of IgM/IgG antibodies against specific Borreliella antigens was analyzed in the presence or absence of clinical manifestations of LB. MATERIAL AND METHODS The study involved 38 patients diagnosed with LB and arthralgia and/or arthritis symptoms, and 57 foresters presenting no clinical symptoms of LB. The ELISA test was applied for general screening of anti-Borreliella IgM/IgG. Western blot was used for confirmatory diagnosis of LB for the positive and borderline results. Serum IL-2, IL-4, IL-6, IL-10, IL-17A, IFN-γ, TNF, IL-8, CCL5, CXCL9/MIG, CCL2/MCP-1, CXCL10/IP-10 concentrations were measured with the use of the Human Cytometric CBA test. The concentration of MMP-2 and MMP-9 in the serum was determined with the use of ELISA tests. RESULTS Analysis of the cytokines and chemokines revealed that only the concentration of IL-2 was significantly higher (2.4 pg/m; p=0.00641) in patients with LB symptoms than in the seropositive individuals (0.4 pg/ml). The MMP2 concentration was significantly higher (233.3 ng/ml; p=0.00294) in patients with clinical manifestations of LB than in those occupationally exposed to tick bites, but did not have anti-Borreliella antibodies (192.0 ng/ml). CONCLUSIONS The presence of IgG antibodies against a number of Borreliella antigens and the differences in the IL-2 and MMP2 levels in seropositive or seronegative individuals and symptomatic LB patients, may indicate differences in the intensity of the immune response to the infection and, consequently, may induce development of clinical manifestations of the disease in seropositive and seronegative individuals.

Modern medicine, as well as the pharmaceutical, cosmetic and food industries appreciate the value of plant raw materials for their antiseptic, analgesic, anti-inflammatory and soothing effects. Medicinal plants, often found endemically, are often treated as potential sources of new substances with antimicrobial and antifungal activity. The aim of this study was to analyze the significance of selected medicinal and spice plants in terms of their ability to inhibit the growth of microorganisms, including those pathogenic to humans. The significance of active substances contained in selected plant species with high antimicrobial potential including Aloe vera L. and Chelidonium majus L. was described. The antimicrobial activity of plant oils, including those extracted from citrus and spice plants, was also analyzed. Plants are a rich source of many substances with a broad spectrum of activity. This knowledge has often been used in folk medicine, but it was not until the development of modern research methods that the mechanisms underlying the therapeutic properties of many plant substances and their proper use could be understood. New substances present in plant materials are constantly being discovered that may be of medical interest because of their great potential to be used against microorganisms or because of their antioxidant and anti-cancer properties.

BACKGROUND This study is aimed at evaluating the awareness among hunters and foresters concerning the activities undertaken for the prevention of Borrelia burgdorferi infections. MATERIAL AND METHODS The study group included 236 hunters from Poland and 339 foresters from Ukraine. The study was based on an original questionnaire including questions which concerned the applied methods of removing ticks attached to the skin, the kind of preventive measures undertaken to avoid tick bites, the evaluation of knowledge concerning the prevention of tick-borne diseases, as well as the participants' willingness to be educated on this issue. RESULTS Removing a tick with a pair of tweezers by a simple vertical move was more frequently practiced by the hunters (16.5%). Spreading an oily substance over a tick to make it go out on its own was more often applied by the foresters (18.8%). The occurrence of erythema migrans was more frequently reported by the foresters (47.2%). Among the persons declaring no knowledge of the prevention of tick-borne diseases, 77.8% of the hunters and 61.5% of the foresters were willing to gain such knowledge, and 11.1% of the hunters and 21.2% of the foresters did not want any education on this issue. The hunters more often than the foresters applied repellents and checked their body after returning from green areas. CONCLUSIONS Education is needed in certain groups undertaking hobby or vocational activities in forest areas of Poland and Ukraine. It seems reasonable to undertake research on the preferences concerning the choice of repellents among the groups at risk of tick bites. Med Pr. 2020;71(1):59-68.

Introduction and objective The incidence of Lyme borreliosis (LB) is increasing in many countries in Europe, including Poland and Slovakia. The aim of the study was to analyze the exposure to tick bites and undertaking LB prevention activities among students of medical fields of studies in Poland (PL) and Slovakia (SK). Material and methods The study was conducted among 611 students from Poland (296 students) and Slovakia (315 students). The applied research tool was the questionnaire. It consisted of questions about exposure to ticks (occurrence of an episode of tick bites, bite site, methods used for tick removal) and the frequency of undertaking preventive actions (using repellents, checking the body after returning from green areas). Pearson Chi-square statistics were calculated to assess significant differences between students from the study countries PL/SK and gender in each country. Results Among surveyed students the episode of a tick bite was reported by 352 persons (57.6%). Students from PL most frequently removed ticks with tweezers with a swift, steady movement (26.6%), while students from SK removed the tick by applying a fatty substance so that it would come off by itself (30.1%). Most of the surveyed students, being outdoors, did not apply ticks repellents (34.7%) or used them rarely (48.9%). At the same time, 49.4% of students stated that they always checked the body to search for any attached tick after returning from green areas. Conclusions Considering the fact that the surveyed people were the students of medical fields of studies, and in the future became qualified medical personnel, the frequency of using the analyzed methods of prophylaxis of LB appears to be too small. The results obtained can help in the education of students of medical fields.

Introduction Lyme borreliosis/Lyme disease is caused by Borrelia burgdorferi and is one of the most common vector-borne diseases transmitted by ticks. Material and Methods A total of 136 Ixodes ricinus ticks, collected in the Ternopil (Ukraine) region, including 126 adults (70 females and 56 males), and 10 nymphs were examined. The identification of the species and their developmental form was based on morphological characteristics. Results PCR with B5S-Bor and 23S-Bor primers resulted in Borrelia burgdorferi sensu lato DNA amplification among six ticks (4.4%). The detailed analysis based on the DNA sequencing showed the presence of DNA of Borrelia afzelii in four samples; the remaining two represented Borrelia burgdorferi sensu lato complex, although their genospecies were not determined. The research confirmed the dominance of Borrelia afzelii genospecies in the ticks from Ukraine. Conclusion It seems reasonable to undertake similar research in ticks from other regions of Ukraine. Knowledge in this field can be useful for public health and planning the prevention of tick-borne diseases.

The aim of our study was the collection of seroprevalence data for Toxoplasma gondii, Coxiella burnetii, Trichinella spp., and Francisella tularensis from hunters in Lublin Province. The antibodies against T. gondii and C. burnetii were recorded in 38.5% and 16.2% of the sera, respectively. 4.05% of the sera were seropositive for both T. gondii and C. burnetii. None of the sera tested reacted positively with F. tulariensis or Trichinella spp. Seroprevalence of T. gondii and C. burnetii is common among the hunters from Lublin Province. It seems reasonable to undertake similar research among hunters from other regions of eastern Poland.

Background. Lyme borreliosis is the most frequent tick-borne disease in Europe and North America, and the number of registered cases is on the increase. Frequent presence in the habitats of ticks enhances the risk of tick bites and possible infection with Borrelia burgdorferi spirochetes. Objective. The aim of the study was to assess the risk of B. burgdorferi infection posed to hunters and other individuals exposed to activity-related contact with ticks. Material and methods. The study was carried out in the northern part of the Lublin Province (eastern Poland) and involved 150 individuals exposed to tick bites (110 hunters and 40 individuals exposed to activity-related contact with ticks). The analysis of sera for the presence of B. burgdorferi IgM and IgG antibodies was carried out. All 150 individuals were tested with the ELISA assay, and positive and borderline results of the assay were verified with the Western blot test. All study participants completed a questionnaire, which provided information about exposure to ticks, application of prophylactic measures, and awareness of Lyme borreliosis. Results. The ELISA assay revealed a positive or borderline result in at least one of the classes of B. burgdorferi antibodies in 63.3% (95/150) of the individuals (IgM 14.0%, IgG 63.3%). Verification carried out with the Western blot test showed a positive or borderline result in at least one of the antibody classes in 38.0% (57/150) of the examined persons (IgM 2.7%, IgG 36.7%). Abdomen (56.0%) and legs (53.7%) were the most frequently bitten body regions. Tick bites on the abdomen were significantly more frequently declared by hunters. Inspection of the body after returning from natural areas was more popular prophylactic method than use of repellents. Inspection of the body was significantly more often used in the group of the hunters. Conclusions. The risk of B. burgdorferi infection among hunters and other individuals undertaking activities associated with exposure to tick bites in the study area is high.

Introduction: Although HEV infection in pigs does not pose a major economic risk to pork production, the risk of zoonotic transmission to humans is an important aspect of public health. HEV genotype 3 infections were reported in developed countries in individuals who had consumed raw meat or meat products from deer, wild boars, or pigs. The aim of the study was the analysis of the occurrence of HEV-specific antibodies among wild boars and domestic pigs in Poland. Material and Methods: A total of 290 samples from wild boars and 143 samples from pigs were tested. The antibodies were tested by ELISA. Results: The presence of anti-HEV IgG was demonstrated in 44.1% of pigs and 31.0% of wild boars. Anti-HEV IgG antibodies were detected in 1.4% of samples from pigs and in 2.1% of samples from wild boars at borderline level. The statistical analysis shows significant differences in the positive results for anti-HEV IgG between the groups of pigs and wild boars (P = 0.0263). Conclusion: Regular surveillance of the occurrence of HEV in swine and wild boars should be performed in the future.

International audience; Small molecules containing quinone and/or oxime moieties have been found as promising anti-fungal agents. One of them is 4-AN, a recently reported potent anti-Candida compound, which inhibits the formation of hyphae, decreases the level of cellular phosphoproteome, and finally shows no toxicity towards human erythrocytes and zebrafish embryos. Here, further research on 4-AN is presented. The results revealed that the compound: (i) Kills Candida clinical isolates, including these with developed antibiotic resistance, (ii) affects mature biofilm, and (iii) moderately disrupts membrane permeability. Atomic force microscopy studies revealed a slight influence of 4-AN on the cell surface architecture. 4-AN was also shown to inhibit multiple various protein kinases, a characteristic shared by most of the ATP-competitive inhibitors. The presented compound can be used in novel strategies in the fight against candidiasis, and reversible protein phosphorylation should be taken into consideration as a target in designing these strategies.

Purpose. In this study, we applied various assays to find new activities of 1,4-naphthoquinone derivatives for potential anti-Candida albicans applications. Methodology. These assays determined (a) the antimicrobial effect on growth/cell multiplication in fungal cultures, (b) the effect on formation of hyphae and biofilm, (c) the influence on cell membrane integrity, (d) the effect on cell morphology using atomic force microscopy, and (e) toxicity against zebrafish embryos. We have demonstrated the activity of these compounds against different Candida species and clinical isolates of C. albicans. Key findings. 1,4-Naphthoquinones significantly affected fungal strains at 8–250 mg l−1 of MIC. Interestingly, at concentrations below MICs, the chemicals showed effectiveness in inhibition of hyphal formation and cell aggregation in Candida. Of note, atomic force microscopy (AFM) analysis revealed an influence of the compounds on cell morphological properties. However, at low concentrations (0.8–31.2 mg l−1), it did not exert any evident toxic effects on zebrafish embryos. Conclusions. Our research has evidenced the effectiveness of 1,4-naphthoquinones as potential anti-Candida agents.

Background Atomic force microscopy (AFM) is an experimental technique which recently has been used in biology, microbiology, and medicine to investigate the topography of surfaces and in the evaluation of mechanical properties of cells. The aim of this study was to evaluate the influence of the complement system and specific anti-Borrelia antibodies in in vitro conditions on the modification of nanomechanical features of B. burgdorferi B31 cells. Material and methods In order to assess the influence of the complement system and anti-Borrelia antibodies on B. burgdorferi s.s. B31 spirochetes, the bacteria were incubated together with plasma of identified status. The samples were applied on the surface of mica disks. Young's modulus and adhesive forces were analyzed with a NanoScope V, MultiMode 8 AFM microscope (Bruker) by the PeakForce QNM technique in air using NanoScope Analysis 1.40 software (Bruker). Results/conclusion The average value of flexibility of spirochetes' surface expressed by Young's modulus was 10185.32 MPa, whereas the adhesion force was 3.68 nN. AFM is a modern tool with a broad spectrum of observational and measurement abilities. Young's modulus and the adhesion force can be treated as parameters in the evaluation of intensity and changes which take place in pathogenic microorganisms under the influence of various lytic factors. The visualization of the changes in association with nanomechanical features provides a realistic portrayal of the lytic abilities of the elements of the innate and adaptive human immune system.

The presence of co-infections induced by tick-borne pathogens in humans is an important epidemiological phenomenon. This issue has attracted growing attention of doctors and people working under conditions of an increased risk of being exposed to tick bites.The research group consisted of 93 individuals with current anti-immunoglobulin M/G (IgM/ IgG) Borrelia burgdorferi or IgG anti-Anaplasma phagocytophilum. The respondents were identified during the screening survey in a group of farmers and foresters occupationally exposed to tick bites. The aim of the work was to analyse the frequency of antibodies to specific antigens of B. burgdorferi and the levels of cytokines in forestry workers and farmers with B. burgdorferi monoinfections and B. burgdorferi / A. phagocytophilum co-infections. Statistical analysis was performed using the Chi2, Mann-Whitney U and Kruskal-Wallis tests.There is a stronger generation of IgG antibodies to B. burgdorferi antigens in patients with B. burgdorferi / A. phagocytophilum co-infections, such as variable major protein-like sequence expressed (VlsE) (p0.05), p19 (p0.02), p17 (p0.05) and complement regulator-acquiring surface protein 3 (CRASP3) (p0.02) compared to persons with B. burgdorferi monoinfections. The discrepancies in the synthesis of cytokines interleukin 6 (IL-6), IL-10, and tumor necrosis factor α (TNF-α) have not been found in persons with B. burgdorferi monoinfections and B. burgdorferi / A. phagocytophilum co-infection.The immune response directed against B. burgdorferi is stronger in patients co-infected with B. burgdorferi and A. phagocytophilum than in those with monoinfection.Występowanie u ludzi koinfekcji patogenami przenoszonymi przez kleszcze jest istotnym zjawiskiem epidemiologicznym, któremu coraz więcej uwagi poświęcają zarówno lekarze, jak i osoby pracujące w warunkach zwiększonego ryzyka ukłucia przez kleszcze.Grupa badana obejmowała 93 osoby z obecnymi przeciwciałami immunoglobuliny M/G (IgM/IgG) anty-Borrelia burgdorferi i IgG anty-Anaplasma phagocytophilum, wyłonione podczas badań przesiewowych z grupy rolników i leśników zawodowo narażonych na pokłucia przez kleszcze. Celem pracy była ocena częstości występowania przeciwciał IgM/IgG dla specyficznych antygenów B. burgdorferi oraz poziomu wybranych cytokin u rolników i leśników w zależności od obecności monoinfekcji B. burgdorferi lub współzakażenia B. burgdorferi / A. phagocytophilum. Uzyskane wyniki badań poddano analizie statystycznej (test Chi2, Manna-Whitneya, Kruskala-Wallisa).U osób z koinfekcją B. burgdorferi i A. phagocytophilum istnieje silniejsze generowanie przeciwciał IgG dla antygenów B. burgdorferi, takich jak VlsE (variable major protein- like sequence expressed) (p0,05), p19 (p0,02), p17 (p0,05) i CRASP3 (complement regulator-acquiring surface protein 3) (p0,02) w porównaniu z osobami z monoinfekcją B. burgdorferi. Nie stwierdza się różnic w poziomie generowanych cytokin interleukiny 6 (IL-6), IL-10, czynnika martwicy nowotworu α (tumor necrosis factor α – TNF-α) u osób z monoinfekcją B. burgdorferi i koinfekcją B. burgdorferi / A. phagocytophilum.U pacjentów z jednoczesnym zakażeniem B. burgdorferi i A. phagocytophilum odpowiedź immunologiczna skierowana przeciwko B. burgdorferi jest silniejsza niż w przypadku monoinfekcji. Med. Pr. 2015;66(5):645–651.

The aim of the present work was to evaluate the topography of spirochetes’ cells Borrelia burgdorferi s.s. B31 in atomic force microscope (AFM). Results: The length of spirochetes B. burgdorferi has ranged between 15,38-22,68µm. The cells of spirochetes do not constitute structures of a fixed diameter and height. Thus, in order to identify real parameters of cells, the horizontal distance and vertical distance have been used in the measurements. The average value of a spirochetes’ diameter has been estimated by taking series of measures and it is 0,40 μm. The average value of a spirochetes’ height has been estimated by taking series of measures and it is 70,14 nm. The analysis of a relation between measured parameters of spirochetes: diameter and height revealed that along with the growth of diameter of a bacteria cell, its height also grows. The average value of a fibers’ diameter has been estimated by taking series of measurements and it is 0,09 μm and the average height of fibers was 7,91 nm. Conclusions: The atomic force microscope (AFM) is a modern tool with a broad spectrum of observatory and measure abilities and is a technique which has been used in biology and microbiology to investigate the topography of surface and in the evaluation properties of cells.

The aim of this study was to analyze the seroprevalence of rubella and cytomegalia among young women. The study included 175 healthy women from the Biała Podlaska District, aged 16 to 35 years. Anti-rubella and anti-CMV IgG were determined by ELISA. 172 (98.3%) study subjects tested positive for rubella antibodies, 1 (0.6%) was seroindeterminate and 2 (1.1%) were seronegative. CMV antibodies were detected in 119 (68.0%) participants; the series included also 1 (0.6%) seroindeterminate and 55 (31.4%) seronegative women. The levels of rubella and CMV antibodies were not significantly affected by age, place of residence and educational level of the study subjects.

The aim of the study was the analysis of the occurrence of specific antibodies against HEV among hunters and foresters who are at risk to be exposed. The study group consisted of 210 hunters (23–80 years old) and 60 foresters (22–64 years old). Anti-HEV IgG were present in 3.81% of the samples of the hunters and in 5% of the samples of the foresters. The statistical analysis shows no significant differences in the results anti-HEV IgG between the groups of hunters and foresters (p = 0.5278). Significantly higher anti-HEV IgG titers were found in the older age group (> 55 years old).

Objectives The objective of the study was assessment of risk of infection with Borrelia burgdorferi in the area of southern Podlasie in Poland, near the border with Belarus, by analysis of post-exposure procedure, and evaluation of asymptomatic infection in adolescents bitten by a tick, confirmed by serologic tests. Material and methods The study was conducted among 128 healthy individuals aged 16-20 who declared being bitten by a tick. The level of IgM and IgG class antibodies was determined using the immunoenzymatic test (Borrelia 14 kD + OspC IgM ELISA and Borrelia IgG + VlsE ELISA, DRG Diagnostics). Positive and doubtful results were confirmed using the Western blot method (EUROLINE-WB, EUROIMMUN). Results In the study group, the largest number of respondents (59.4%) declared tick bite in the region of the lower extremities, most often in the knee pit. Among the methods for removing the tick the largest number of respondents indicated removing it with the use of tweezers, with a simple, swift steady movement (29.7%), and pulling it out with the fingers (22.7%). In the ELISA test, a positive or doubtful result in at least one class was observed in 25.0% of respondents (n=32/128): in IgM class - 23.4% (n=30/128), and in IgG class - 4.7% (n=6/128). After verification with the Western blot test, infection was confirmed in 5.5% of respondents (n=7/128): in IgM class - 1.6% (n=2/128), in IgG class - 3.9% (n=5/128). In IgM class antibodies, the Western blot test confirmed positive or doubtful results of the ELISA test in 6.7%, while in IgG class antibodies in 83.3%. Conclusion Evaluation of the actual infection with Borrelia spp. using serologic tests is difficult due to a certain non-specificity of the ELISA test, especially in IgM class antibodies, and difficulties with performance of a wide scope of specific Western blot tests. The variety of methods of tick removal declared by adolescents suggests that a wider education of society concerning appropriate methods of removing the tick should become an especially important element of prophylactic actions in the area of borreliosis.

Summary: The aim of our this work was analysis of speciϐic immune respon se for B. burgdorferi according to in vivo antigens in regard to symptoms subjectively perceived by foresters, in 2009 and 2012. Material and methods: study group include 41 foresters (19 - 86 years old), from the forest district in Lubelskie region. All persons underwent serological diagnostic. We used 2 phase diagnosis method of Lyme diseases according to standards. Used tests include Borrelia antigens from in vivo group, which are not use in standard diagnosis test. The study was carried out in 2009 and then again in 2012. Results: The analysis of antibodies for Borrelia proteins, including the in vivo, demonstrated how active, long-term and time-variable is the process of B. burgdorferi infection. Conclusions: The expression of Borrelia proteins and high immunogenicity translates into effectiveness of diagnostic procedures concerning B. burgdorferi identiϐication. Considering to not clear clinical manifestatio n and changeable immune response require individual approach to each patient.

Background: The aim of the work was to analyze the incidence of infection with Borrelia burgdorferi in forestry workers and farmers, major groups occupationally exposed to tick bites. Material and Methods: The study group included 275 workers (171 foresters and 104 farmers). The control group consisted of 45 people, who have not been occupationally exposed to tick bites. The screening Elisa and Wb tests for the presence of anti-Borrelia IgM/IgG antibodies were performed in all subjects of the study and control groups. Statistical analysis was performed using the Chi2 test. Results: The positive results denoting the presence of anti-Borrelia IgM/IgG antibodies were found in 55% of farmers and 28% of foresters occupationally exposed to Lyme borreliosis and coming from the area of South Podlasie Lowland and Lublin Polesie. The differences between the forestry workers and the control group (p ≤ 0.00001) and between farmers and the control group (p ≤ 0.001) were statistically significant. The species, such as B. spielmanii and B. bavariensis, which have not yet been reported in Poland, are significant etiologic agents of Lyme disease. Conclusion: The risk of occupational exposure to the B. burgdorferi infection is high for foresters and farmers, and the infection with spirochetes is frequently confirmed on the basis of positive results of the Wb test. The presence of specific antibodies against protein antigens of B. spielmanii and B. bavariensis suggest that these bacteria can cause Lyme disease both independently and in participation with other Borrelia species, which influences the development of the clinical manifestations of infection. Med Pr 2014;65(1):109–117