Changes in the Mechanical Properties of Alginate-Gelatin Hydrogels with the Addition of Pygeum africanum with Potential Application in Urology.

New hydrogel materials developed to improve soft tissue healing are an alternative for medical applications, such as tissue regeneration or enhancing the biotolerance effect in the tissue-implant-body fluid system. The biggest advantages of hydrogel materials are the presence of a large amount of water and a polymeric structure that corresponds to the extracellular matrix, which allows to create healing conditions similar to physiological ones. The present work deals with the change in mechanical properties of sodium alginate mixed with gelatin containing Pygeum africanum. The work primarily concentrates on the evaluation of the mechanical properties of the hydrogel materials produced by the sol-gel method. The antimicrobial activity of the hydrogels was investigated based on the population growth dynamics of Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923, as well as the degree of degradation after contact with urine using an innovative method with a urine flow simulation stand. On the basis of mechanical tests, it was found that sodium alginate-based hydrogels with gelatin showed weaker mechanical properties than without the additive. In addition, gelatin accelerates the degradation process of the produced hydrogel materials. Antimicrobial studies have shown that the presence of African plum bark extract in the hydrogel enhances the inhibitory effect on Gram-positive and Gram-negative bacteria. The research topic was considered due to the increased demand from patients for medical devices to promote healing of urethral epithelial injuries in order to prevent the formation of urethral strictures.

Evaluation of biofilm formation on acrylic resins used to fabricate dental temporary restorations with the use of 3D printing technology.

BACKGROUND: Temporary implant-retained restorations are required to support function and esthetics of the masticatory system until the final restoration is completed and delivered. Acrylic resins are commonly used in prosthetic dentistry and lately they have been used in three-dimensional (3D) printing technology. Since this technology it is fairly new, the number of studies on their susceptibility to microbial adhesion is low. Restorations placed even for a short period of time may become the reservoir for microorganisms that may affect the peri-implant tissues and trigger inflammation endangering further procedures. The aim of the study was to test the biofilm formation on acrylamide resins used to fabricate temporary restorations in 3D printing technology and to assess if the post-processing impacts microbial adhesion. METHODS: Disk-shaped samples were manufactured using the 3D printing technique from three commercially available UV-curable resins consisting of acrylate and methacrylate oligomers with various time and inhibitors of polymerization (NextDent MFH bleach, NextDent 3D Plus, MazicD Temp). The tested samples were raw, polished and glazed. The ability to create biofilm by oral streptococci (S. mutans, S. sanguinis, S. oralis, S. mitis) was tested, as well as species with higher pathogenic potential: Staphylococcus aureus, Staphylococcus epidermidis and Candida albicans. The roughness of the materials was measured by an atomic force microscope. Biofilm formation was assessed after 72 h of incubation by crystal violet staining with absorbance measurement, quantification of viable microorganisms, and imaging with a scanning electron microscope (SEM). RESULTS: Each tested species formed the biofilm on the samples of all three resins. Post-production processing resulted in reduced roughness parameters and biofilm abundance. Polishing and glazing reduced roughness parameters significantly in the NextDent resin group, while glazing alone caused significant surface smoothing in Mazic Temp. A thin layer of microbial biofilm covered glazed resin surfaces with a small number of microorganisms for all tested strains except S. oralis and S. epidermidis, while raw and polished surfaces were covered with a dense biofilm, rich in microorganisms. CONCLUSIONS: UV-curing acrylic resins used for fabricating temporary restorations in the 3D technology are the interim solution, but are susceptible to adhesion and biofilm formation by oral streptococci, staphylococci and Candida. Post-processing and particularly glazing process significantly reduce bacterial biofilm formation and the risk of failure of final restoration.

[Well-known and new variants of pathogenic Escherichia coli as a consequence of the plastic genome]. / Znane i nowe warianty patogennych Escherichia coli jako konsekwencja plastycznego genomu.

E. coli is a diverse bacterial species encompassing commensal as well as intestinal and extraintestinal pathogenic strains. The ability to adapt to so many different niches in the host organism is determined by the extreme genomic plasticity of E. coli. The genetic diversity is due to a complex phylogenetic structure in which besides the well-known main groups A, B1, B2 and D, four new groups, C, E, F and Clad I, have been characterized recently. The mobile gene pool exchanged by horizontal gene transfer (HGT) is another important driving force in the evolution of E. coli. Pathogenicity of strains is conditioned by a specific repertoire of virulence factors located on the mobile genetic elements and transmitted by HGT. The environment changing constantly stimulates the formation of new virulence gene combinations that generate the formation, not observed so far, of new pathogenic clones of higher capacity for virulence and greater expansiveness. The presence of very similar virulence plasmids carrying conserved combinations of the virulence genes (CVP) among extraintestinal pathogenic strains in humans and birds has been observed. It indicates a real possibility of occurrence of common virulence factors. The increase in drug resistance among pathogenic E. coli is also reflected in the prevalence of highly expansive clones exhibiting both high virulence and resistance. The presented data indicate that further studies are required to determine the interdependencies of resistance and virulence at the genetic level to help improve our management of the infectious diseases caused by these bacteria.

Age as a factor influencing diversity of commensal E. coli microflora in pigs.

Commensal, intestinal E. coli microflora plays a role in maintenance of intestinal balance of the host, is responsible for defending against pathogenic E. coli. This study encompasses the analysis of BOX-PCR fingerprinting patterns, phylogenetic grouping and virulence genes prevalence among commensal E. coli isolates derived from healthy pigs. Altogether, 274 unique E. coli isolates were identified, 110 from weaned piglets (Piglets I and Piglets II) and 164 from adult sows (Sows I and Sows II). BOX-PCR analysis distinguished isolates from pigs in different age and indicated that during maturation the changes in E. coli microflora occurred. Phylogenetic grouping revealed significant differences between distribution of four phylogenetic groups among isolates derived from piglets and sows. In phylogenetic structure of isolates from the piglets group B1 prevailed significantly, while among isolates derived from the sows the majority of them were classified into phylogenetic group A. The identification of 17 virulence factors in E. coli isolates derived from healthy pigs was performed. Three of 13 intestinal (escV, ehxA, estII) and four extra-intestinal virulence genes (VGs) (hlyA, fimH, papA, sfaS) were detected in the porcine isolates. The percentage of VGs positive isolates among piglets is higher than among sows, moreover, the VGs occurring in E. coli isolates from piglets revealed greater diversity than that detected among isolates from sows.

The phenotypic and genotypic characteristics of antibiotic resistance in Escherichia coli populations isolated from farm animals with different exposure to antimicrobial agents.

The aim of the study was to determine the influence of the presence or the absence of antibiotic input on the emergence and maintenance of resistance in commensal bacteria from food producing animals. The research material constituted E. coli isolates from two animal species: swine at different age from one conventional pig farm with antibiotic input in young pigs and from beef and dairy cattle originated from organic breeding farm. The sensitivity to 16 antimicrobial agents was tested, and the presence of 15 resistance genes was examined. In E. coli from swine, the most prevalent resistance was resistance to streptomycin (88.3%), co-trimoxazole (78.8%), tetracycline (57.3%) ampicillin (49.3%) and doxycycline (44.9%) with multiple resistance in the majority. The most commonly observed resistance genes were: bla(TEM) (45.2%), tetA (35.8%), aadA1 (35.0%), sul3 (29.5%), dfrA1 (20.4%). Differences in phenotypes and genotypes of E. coli between young swine undergoing prevention program and the older ones without the antibiotic pressure occurred. A disparate resistance was found in E. coli from cattle: cephalothin (36.9%), cefuroxime (18.9%), doxycycline (8.2%), nitrofurantoin (7.7%), and concerned mainly dairy cows. Among isolates from cattle, multidrug resistance was outnumbered by resistance to one or two antibiotics and the only found gene markers were: bla(SHV), (3.4%), tetA (1.29%), bla(TEM) (0.43%) and tetC (0.43%). The presented outcomes provide evidence that antimicrobial pressure contributes to resistance development, and enteric microflora constitutes an essential reservoir of resistance genes.

Personalized, 3D- printed fracture fixation plates versus commonly used orthopedic implant materials- biomaterials characteristics and bacterial biofilm formation.

Additive manufacturing enabled the development of personalized, ideally fitting medical devices. The topography of the surface of the 3D-printed implant may not only facilitate its integration but also cause its rejection, as the surface may become a reservoir for different bacterial strains. In this study, the innovative, raw, 3D- printed fracture fixation plates, manufactured by using selective laser melting (SLM) from Ti-6Al-4V were compared with commercially available, surface-modified plates commonly used in orthopedic surgery. The topography surface of the plates was studied by atomic force microscopy. Susceptibility to the development of biofilm was tested for Staphylococcus epidermidis, Staphylococcus aureus and Streptococcus mutans by using crystal violet staining of biomass, confocal, and scanning electron microscopy (SEM). 3D- printed plates showed higher roughness (Sa=131.0 nm) than commercial plates (CP1 and CP2), Sa= 60.67 nm and Sa=55.48 nm, respectively. All strains of bacteria colonized 3D- printed raw plates more densely than commercial plates. The microscopic visualization showed biofilm mostly in irregular cavities of printed plates while on commercial plates it was mainly located along the edges. The research has indicated that there is need for further development of this technology to optimize its effectiveness and safety.

MicroRNA-Based Fingerprinting of Cervical Lesions and Cancer.

The regulatory functions of microRNA (miRNA) are involved in all processes contributing to carcinogenesis and response to viral infections. Cervical cancer in most cases is caused by the persistence of high-risk human papillomavirus (HR-HPV) infection. While oncogenic human papillomaviruses induce aberrant expression of many cellular miRNAs, this dysregulation could be harnessed as a marker in early diagnosis of HR-HPV infection, cervical squamous intraepithelial lesions, and cancer. In recent years, growing data indicate that miRNAs show specific patterns at various stages of cervical pathology. The aim of this review is to systematize current reports on miRNA capacity that can be utilized in personalized diagnostics of cervical precancerous and cancerous lesions. The analysis of the resources available in online databases (National Center for Biotechnology Information-NCBI, PubMed, ScienceDirect, Scopus) was performed. To date, no standardized diagnostic algorithm using the miRNA pattern in cervical pathology has been defined. However, the high sensitivity and specificity of the reported assays gives hope for the development of non-invasive diagnostic tests that take into account the heterogeneity of tumor-related changes. Due to this variability resulting in difficult to predict clinical outcomes, precise molecular tools are needed to improve the diagnostic and therapeutic process.

Antibacterial Activity of Bacteriocinogenic Commensal Escherichia coli against Zoonotic Strains Resistant and Sensitive to Antibiotics.

Antibiotic resistance concerns various areas with high consumption of antibiotics, including husbandry. Resistant strains are transmitted to humans from livestock and agricultural products via the food chain and may pose a health risk. The commensal microbiota protects against the invasion of environmental strains by secretion of bacteriocins, among other mechanisms. The present study aims to characterize the bactericidal potential of bacteriocinogenic Escherichia coli from healthy humans against multidrug-resistant and antibiotic-sensitive strains from pigs and cattle. Bacteriocin production was tested by the double-layer plate method, and bacteriocin genes were identified by the PCR method. At least one bacteriocinogenic E. coli was detected in the fecal samples of 55% of tested individuals, adults and children. Among all isolates (n = 210), 37.1% were bacteriocinogenic and contained genes of colicin (Col) Ib, ColE1, microcin (Mcc) H47, ColIa, ColM, MccV, ColK, ColB, and single ColE2 and ColE7. Twenty-five E. coli carrying various sets of bacteriocin genes were further characterized and tested for their activity against zoonotic strains (n = 60). Strains with ColE7 (88%), ColE1-ColIa-ColK-MccH47 (85%), MccH47-MccV (85%), ColE1-ColIa-ColM (82%), ColE1 (75%), ColM (67%), and ColK (65%) were most active against zoonotic strains. Statistically significant differences in activity toward antibiotic-resistant strains were shown by commensal E. coli carrying MccV, ColK-MccV, and ColIb-ColK. The study demonstrates that bacteriocinogenic commensal E. coli exerts antagonistic activity against zoonotic strains and may constitute a defense line against multidrug-resistant strains.

Susceptibility to biofilm formation on 3D-printed titanium fixation plates used in the mandible: a preliminary study.

Background: In the oral and maxillofacial surgery, fixation plates are commonly used for the stabilization of bone fragments. Additive manufacturing has enabled us to design and create personalized fixation devices that would ideally fit any given fracture. Aim: The aim of the present preliminary study was to assess the susceptibility of 3D-printed titanium fixation plates to biofilm formation. Methods: Plates were manufactured using selective laser melting (SLM) from Ti-6Al-4 V. Reference strains of Streptococcus mutans, Staphyloccocus epidermidis, Staphylococcus aureus, Lactobacillus rhamnosus, and Candida albicans, were tested to evaluate the material's susceptibility to biofilm formation over 48 hours. Biofilm formations were quantified by a colorimetric method and colony-forming units (CFU) quantification. Scanning electron microscopy (SEM) visualized the structure of the biofilm. Results: Surface analysis revealed the average roughness of 102.75 nm and irregular topography of the tested plates. They were susceptible to biofilm formation by all tested strains. The average CFUs were as follows: S. mutans (11.91 x 107) > S.epidermidis (4.45 x 107) > S. aureus (2.3 x 107) > C.albicans (1.22 x 107) > L. rhamnosus (0.78 x 107). Conclusions: The present preliminary study showed that rough surfaces of additively manufactured titanium plates are susceptible to microbial adhesion. The research should be continued in order to compare additively manufactured plates with other commercially available osteotomy plates. Therefore, we suggest caution when using this type of material.

Evaluation of the Antibacterial Activity of Ag- and Au-Nanoparticles Loaded TiO2 Nanotubes.

Current research on the antibacterial properties of implant surfaces has focused on using titanium nanotubes (TNTs) with diameters of 100 and 200 nm, which simultaneously show the best antibacterial properties, poor osseointegration, and ability to immobilize proteins. Therefore, the research aimed to develop an implantable material based on titanium dioxide nanotubes with a diameter of 50 nm doped with silver (AgNPs) and gold nanoparticles (AuNPs), indicating good absorption and antibacterial properties. Moreover, metallic nanoparticles deposited by varying methods should maintain sphericity and lack of agglomeration. For this purpose, the surface charge, wettability, stability of nanoparticles, and antibacterial properties against Gram-positive and Gram-negative bacteria, i. e., Staphylococcus epidermidis, Streptococcus mutans, and Pseudomonas aeruginosa , were performed. Obtained results indicate a greater resistance to leaching of silver nanoparticles compared to gold nanoparticles. These results are reflected in microbiological studies, both into the time and the effectiveness of the implantable material's antibacterial activity. A greater antibacterial effect of AgNPs than AuNPs has been confirmed. Also, AgNPs inhibit the multiplication of Gram-negative bacteria to a greater extent than Gram-positive bacteria. It has been proven that the TNT platforms deposited with metal nanoparticles via the voltammetric method are more effective in deactivating microorganisms. Besides, the results have proven that smaller TNTs effectively reduce live bacteria as nanotubes with a diameter of 100 and 200 nm.

Extended Phylogeny and Extraintestinal Virulence Potential of Commensal Escherichia coli from Piglets and Sows.

Commensal Escherichia coli, naturally occurring in the intestinal tract, can be the origin of extraintestinal pathogenic E. coli (ExPEC) strains. ExPEC causes high mortality and significant economic losses in the swine industry in several countries and poses a serious threat to public health worldwide. The aim of this study was to analyze the extended phylogenetic structure and extraintestinal virulence potential in two groups of commensal E. coli isolates from post-weaning piglets and sows. The phylogenetic assignment to eight groups was determined using the revised Clermont phylogenetic typing method in quadruplex PCR. Identification of extraintestinal virulence genes (VGs) and adhesin operon genes was performed using multiplex or simplex PCR. The revised phylogenetic assignment allowed us to distinguish E. coli with significantly higher (groups C and F) or lower (group E) virulence potential in isolates from piglets. The majority of the tested VGs occurred more frequently in isolates from piglets than from sows, with statistically significant differences for seven genes: fimH, papAH, iutA, iroN, ompT, traT, and iss. Complete operons for type I and P fimbriae significantly prevailed among E. coli from piglets. This study provides insight into the extended phylogenetic structure of porcine commensal E. coli and showed that these strains, particularly from piglets, constitute a considerable reservoir of extraintestinal VGs and may increase the potential risk of extraintestinal infections.

Phylogenetic background, virulence gene profiles, and genomic diversity in commensal Escherichia coli isolated from ten mammal species living in one zoo.

Three hundred commensal Escherichia coli recovered from healthy herbivorous, carnivorous, and omnivorous mammals from one zoo were characterized for their phylogenetic origin, intestinal virulence gene (VG) prevalence, and genomic diversity. The phylogenetic structure of the E. coli (groups A, B1, B2, and D) from the herbivores was homogenous, with a prevailing representation of group B1. In the carnivores and omnivores, the phylogenetic diversity was species specific with a higher representation of group A compared to the herbivores. Of 16 intestinal VGs in the whole set, 8 were detected and they formed 13 VG profiles. In the herbivores, all the VG-positive isolates belonged to group B1 and harboured the genes eaeA, eastI, ehxA, stx1, and stx2, which separately or in combination formed 8 VG profiles. In the carnivores and omnivores, the VG-positive isolates frequently belonged to group A and harboured the estI and estII genes or a combination of eastI and estI, forming three VG profiles. Single genes cnf2, in group B2, and eastI, in group D, were found. Similarity analysis of pulsed-field gel electrophoresis (PFGE) patterns revealed closer relatedness between the isolates from carnivores and omnivores than those from herbivores. The comparison between the prevalence of phylogenetic groups and the phylogenetic origin of VG-positive isolates in the examined E. coli suggested, that E. coli from group B1 in herbivores and E. coli from group A rather than B1 in carnivores and omnivores are "best adapted" to the host organism. The groups revealed different preferences in the acquisition and maintenance of intestinal VGs.

Complexity of Antibiotic Resistance in Commensal Escherichia coli Derived from Pigs from an Intensive-Production Farm.

Antibiotics in animal husbandry are used to maintain welfare, but lead to the generation of resistant strains. We analyzed commensal multidrug-resistant Escherichia coli from pigs at the beginning and end of the production cycle in a farm with a farrow-to-finish system in order to investigate whether clonal spread or horizontal gene transfer constitutes the main factor responsible for the prevalence of resistance in this environment. Among 380 isolates, 56 multidrug-resistant E. coli with a similar resistant phenotype were selected for more detailed investigations including a genomic similarity analysis and the detection of mobile elements. Isolates carried blaTEM-1, aadA1, strA/B, tetA, tetB, tetC, dfrA1, dfrA5, dfrA7, dfrA12, sul1, sul2, sul3, and qnrS resistance genes, with the common co-occurrence of genes encoding the same resistance phenotype. A pulse-field gel electrophoresis analysis of the genomic similarity of multidrug-resistant E. coli showed ≤65% similarity of most of the tested strains and did not reveal a dominant clone responsible for the prevalence of resistance. Class 1 and 2 integrons and transposons 7 and 21 were detected among mobile elements; however, some were truncated. Plasmids were represented by 11 different incompatibility groups (K, FIB, I1, FIIA, FIC, FIA, Y, P, HI1, B/O, and T). Genetic resistance traits were unevenly spread in the clonal groups and suggested the major rearrangement of genetic material by horizontal gene transfer. The present results revealed that in commensal E. coli from pigs in a homogeneous farm environment, there was no dominant clone responsible for the spread of resistance and persistence in the population.

Comparison of Commensal Escherichia coli Isolates from Adults and Young Children in Lubuskie Province, Poland: Virulence Potential, Phylogeny and Antimicrobial Resistance.

Commensal Escherichia coli population is a dynamic structure which may be important in the pathogenesis of extraintestinal infections. The aim of this study was the comparison of genetic diversity of commensal E. coli isolates from two age group-adults and young children. E. coli strains were isolated on MacConkey agar and identified by biochemical tests. Determination of four major phylogenetic groups, identification of virulence genes and antimicrobial resistance determinants were performed by using multiplex or simplex PCR. Phenotypic analysis of resistance was based on disc-diffusion method. The prevalence of virulence genes was significantly higher among isolates from adults than from young children. Phylogroup B2 predominated among E. coli from adults, whereas phylogroup A was the most common in isolates from young children. The analyses of antimicrobial resistance revealed that resistance to at least one antimicrobial agent and multidrug-resistance were detected significantly more frequent in the isolates from adults than from young children. This study documented that the commensal E. coli isolates from adults showed greater genetic diversity than from young children and constitutes a substantial reservoir of the virulence genes typical for extraintestinal pathogenic E. coli.

Prevalence of antibiotic resistance profile in relation to phylogenetic background among commensal Escherichia coli derived from various mammals.

The paper describes the prevalence of resistant strains within the genetic structure of E. coli (phylogenetic group A, B1, B2 and D). A total of 200 commensal E. coli strains have been derived from 10 species of healthy animals residing on ZOO Safari Park area, in Swierkocin, Poland. The phylogenetic structure of E. coli has been analysed with the use of a PCR-based method. The strains were tested in terms of their susceptibility to eight classes of antibiotics: aminoglycosides, penicillins, cephalosporins, tetracyclines, nitrofurans, sulphonamides, phinicols, and quinolones. The genetic structure of E. coli revealed a not uniform distribution of strains among the four phylogenetic groups with significantly numerous representation of groups A and B1. Resistant E. coli were found within each of the phylogenetic groups. Strains resistant to one class of antibiotics occurred significantly more frequently in phylogenetic groups B2 and D (potential pathogens), whereas strains resistant to more than one class of antibiotics belonged to phylogenetic groups A and B1 (typical commensals) in a prevailing number of cases.

Prevalence of virulence determinants and antimicrobial resistance among commensal Escherichia coli derived from dairy and beef cattle.

Cattle is a reservoir of potentially pathogenic E. coli, bacteria that can represent a significant threat to public health, hence it is crucial to monitor the prevalence of the genetic determinants of virulence and antimicrobial resistance among the E. coli population. The aim of this study was the analysis of the phylogenetic structure, distribution of virulence factors (VFs) and prevalence of antimicrobial resistance among E. coli isolated from two groups of healthy cattle: 50 cows housed in the conventional barn (147 isolates) and 42 cows living on the ecological pasture (118 isolates). The phylogenetic analysis, identification of VFs and antimicrobial resistance genes were based on either multiplex or simplex PCR. The antimicrobial susceptibilities of E. coli were examined using the broth microdilution method. Two statistical approaches were used to analyse the results obtained for two groups of cattle. The relations between the dependent (VFs profiles, antibiotics) and the independent variables were described using the two models. The mixed logit model was used to characterise the prevalence of the analysed factors in the sets of isolates. The univariate logistic regression model was used to characterise the prevalence of these factors in particular animals. Given each model, the odds ratio (OR) and the 95% confidence interval for the population were estimated. The phylogroup B1 was predominant among isolates from beef cattle, while the phylogroups A, B1 and D occurred with equal frequency among isolates from dairy cattle. The frequency of VFs-positive isolates was significantly higher among isolates from beef cattle. E. coli from dairy cattle revealed significantly higher resistance to antibiotics. Some of the tested resistance genes were present among isolates from dairy cattle. Our study showed that the habitat and diet may affect the genetic diversity of commensal E. coli in the cattle. The results suggest that the ecological pasture habitat is related to the increased spreading rate of the VFs, while the barn habitat is characterised by the higher levels of antimicrobial resistance among E. coli.

Antimicrobial resistance in commensal Escherichia coli from pigs during metaphylactic trimethoprim and sulfamethoxazole treatment and in the post-exposure period.

The prevalence of trimethoprim (TMP) and sulfamethoxazole (SMX) resistance in commensal E. coli from pigs was tested in this study. E. coli was derived from three groups of piglets in successive stages of metaphylactic therapy and from two groups of sows 10 and 18 weeks after the treatment. MIC values of TMP and SMX were determined for a total of 352 strains. The presence of resistance genes (dfrA1, dfrA5, dfrA7, dfrA12, dfrA17, sul1, sul2, sul3) and class 1 and 2 integron-associated dfrA gene cassettes was tested. Resistance to TMP was very high during the administration of the antimicrobial (from 97 to 100%) and amounted to 86% and 69% in the post-exposure period; MIC > 32 mg/L. The isolates from all groups of pigs were resistant to sulfamethoxazole, with MIC > 1028 mg/L. The dfrA1 and sul1 genes (as part of integrons) dominated in E. coli from piglets, but the dfrA12 and sul1 genes were prevalent in E. coli from sows. Coexistence of the different dfrA genes was detected in 71 isolates from all groups of swine. Transcription analysis revealed that most of these genes were not transcribed, particularly gene cassettes of class 1 integrons. The research revealed a high level of resistance associated with the metaphylactic treatment, persistence and circulation of resistance in bacterial populations. Diverse genetic background with multiple and not transcribed resistance genes was observed.

The influence of hypoxic physical activity on cfDNA as a new marker of vascular inflammation.

The phenomenon of circulating cell-free DNA (cfDNA) is important for many biomedical disciplines including the field of exercise biochemistry and physiology. It is likely that cfDNA is released into the plasma by apoptosis of endothelial cells and circulating endothelial progenitor cells (EPCs), and/or by NETosis of immune cells induced by strenuous exercise. Increases of cfDNA are described to be a potential hallmark for the overtraining syndrome, and might be related to aseptic vascular inflammation in athletes. Yet, the relevance of systemic inflammation and cfDNA with endothelial dysfunction in athletes still remains unclear. In this review article, we provide a current overview of exercise-induced cfDNA release to the circulation with special emphasis on its relationship with apoptosis and NETosis and the effect of hypoxic physical activity on vascular inflammation in athletes.

Type 1 fimbriae in commensal Escherichia coli derived from healthy humans.

Type 1 fimbriae are one of the most important factors of Escherichia coli adaptation to different niches in the host. Our study indicated that the genetic marker--fimH gene occurred commonly in commensal E. coli derived from healthy humans but expression of the type 1 fimbriae was not observed. Identification of fim structural subunit genes (fimA-fimH) and recombinase fimE and fimB genes showed that many of the strains were carrying an incomplete set of genes and the genes expression study revealed that in strains with complete set of fim genes, the fimC gene, encoding the chaperone protein, was not expressed.

(AC)n microsatellite polymorphism and 14-nucleotide deletion in exon 42 ankyrin-1 gene in several families with hereditary spherocytosis in a population of South-Western Poland.

Defects in ankyrin-1 have been implicated in approximately half of all patients with hereditary spherocytosis. However, not all polymorphisms in this gene lead to the changes in expressed protein or to the changes of the level of its expression. In this study, we report on several cases of the (AC)n microsatellite polymorphism in 3' untranslated region of ANK1 gene found in nine families (19 patients) with hereditary spherocytosis (HS) and also in ten healthy individuals from the same territory. We also found that 14-nucleotide deletion in this region of ANK1 which was shifted by five nucleotides in relation to another 14-nucleotide deletion listed in Single Nucleotide Polymorphism National Center for Biotechnology Information (SNP NCBI) database. This deletion seems to be present only in individuals with 11/14 and 13/14 AC repeats what would be an interesting correlation between these two features. However, comparison of the data obtained for HS patients and healthy individuals indicates that both polymorphisms are not connected to the pathology of hereditary spherocytosis.