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The paper presents determinants of location of the multicultural complex of archaeological sites in the vicinity of Ulów, in the Central Roztocze upland region in south-eastern Poland. Archaeological research revealed that in the area assumed to be devoid of settlements, the settlements of prehistoric and historical communities functioned from the Palaeolithic to modern times. The region was also subjected to environmental examination. Location of sites was analysed, taking into account a convenience of communication in a regional scale and local environmental conditions. Analysis of hydrogeological, geomorphological and soil conditions was carried out, taking into account water supply, communication and the farming development. In-depth analysis included micromorphological DTM (Digital Terrain Model) and geological and soil probing. The area was found to be located on the crossing of prehistoric communication routes the course of which depended on the variability of the physiographic parameters of regions. The functioning of new cultures in the same place resulted from specific local conditions such as: easily arable soils, favourable microclimate, and particularly access to water. The presence of a source of water in a plateau area is determined tectonically (strike-slip fault), lithologically (impermeable marl horizon), and geomorphologically (dissection of the aquifer by an erosion-denudation valley).

Introduction.Type 2 diabetes is a chronic disease with an epidemic character. Its prevalence is associated with lifestyle, many environmental factors, and genetic determinants. Implementation of diet therapy is the basis for treatment of the disease.

The aim of the study was to determine the mechanical and geometric properties as well as bone tissue density of long bones in primiparous and multiparous dams of minks supplemented with β-hydroxy β-methylbutyrate (HMB) and/or 2-oxoketoglutarate (2-Ox) during gestation. Powdered 2-Ox was given at the daily dosage of 0.4 g/kg b.w. separately or simultaneously with HMB, which was administered at the daily dosage of 0.02 g/kg b.w. The study demonstrates for the first time that administration of 2-Ox and/or HMB to dams markedly influences bone tissue density and the mechanical and geometrical properties of mother`s bones in minks. Moreover, it was demonstrated that the supplementation was more effective in the thoracic limb, which was comprehensively used in contrast to the pelvic limb. The mechanical parameters and bone tissue density significantly increased in the humerus in multiparous minks. Only such diet may provide satisfactory production results in the animals. Nutritional deficiencies occurring during pregnancies may trigger body`s own reserves to cover the bone mass increase in developing foetuses and support milk production. This can prevent regeneration of dams’ organisms, which negatively affects their reproductive performance. 2-Ox or HMB may be regarded as a protective metabolite when administered orally to minks, counteracting the negative influences of pregnancy and lactation periods on bones condition. Both simultaneous treatment with 2-Ox and HMB and their separate administration were equally effective.

The aim of this study was to detect possible differences between farmed and wild-living raccoon dogs. Analysis of polymorphism in 15 microsatellite sequences led to the conclusion that raccoon dogs raised on Polish farms and wild raccoon dogs living in Poland are two genetically distinct groups of animals. Wild Polish raccoon dogs are genetically more similar to the population of wild animals from the Kaliningrad Region than to farmed animals. The analysis of microsatellite loci showed clear genetic differences between farmed and wild-living populations of raccoon dog, despite only 50 years of isolation of the two groups of animals. The farmed population was characterized by higher genetic variation than the wild-living population. On the basis of the analyses three microsatellite loci (INU014, Ren13J22 and Ren41D20) were proposed for determination of the origin of animals that have escaped from farms.

•The intracellular free iron in E.coli hemHappear to be double that in wt strain.•Increased Fe(II) and AlkB concentrations result in decreased MMS-induced mutations.•Dealkylation of dNTPs takes place in the presence of Fe(II) and not requires AlkB.

Creation of interspecific hybrids is widely common among plants and animals in order to improve economically important traits for humans. The studied material consisted of chromosomal preparations in the metaphase stage obtained from interspecies hybrids of arctic foxes (Alopex

Selected genes were analyzed in silicoin three species: red fox (Vulpes vulpes), raccoon dog (Nyctereutes procyonoides), and dog (Canis lupus familiaris). This type of analysis exemplifies current and potential research on gene expression. Four nucleotide sequences, of the genes IGF1, MYO15A, PAX3and MC1R, were obtained from the NCBI online database. The analyses focused on the presence of CpG islands and two analytical techniques, BSP and MSP. The results from three computer programs, CpG Island Searcher®, BiSearch®and MethPrimer®, were discussed in detail. The applications were compared in terms of their functionality and usefulness.

Understanding the surface chemistry and reactivity of 2D MXenes is of fundamental importance in developing more advanced functional materials. In this work, we describe multilayered Ti3C2MXene-based materials such as Ti3C2/Al3＋, Ti3C2/In3+and Ti3C2/Ga3+, obtained by modifying the MXene surface with aluminum-, indium- and gallium alkoxides. In the synthesis of these materials, organometallic compounds of group 13 metals (Et3Al, Me3In, and Et3Ga) were chosen as precursors for the alkoxides to modify the surface. To attach organometallic compounds to the MXene surface, their high reactivity towards the terminal OH, = O and F groups present on the MXene surface was used. In the next step, exposure of the surface-attached aluminum-, indium and gallium alkyls to air allowed their transformation into metal alkoxides due to the high reactivity of the metal-carbon bonds to oxygen and water. In our method of MXene modification, the alkoxide precursors can be extended to other organometallic compounds, such as groups 1, 2 and 12 alkyls. Microbiological studies of the developed 2D Ti3C2/Al3＋, Ti3C2/In3+and Ti3C2/Ga3+alkoxides showed no acute ecotoxicity to the tested microorganisms. The obtained materials did not influence bioluminescent/biosensor-type microorganisms nor reduce the viability of other types of microorganisms, such as Staphylococcus aureus, Escherichia colior Candida albicans.The obtained results clearly indicate that the surface modification of 2D Ti3C2MXene with group 13 metal compounds is highly prospective toward the development of various types of materials that do not show any adverse effects to living organisms.

This study was designed to determine the degree of genetic distinctiveness between farmed and wild foxes (Vulpes vulpes). Analysis of polymorphism in 16 microsatellite sequences led to the conclusion that red foxes raised on Polish farms and wild foxes living in Poland are two groups of genetically distinct animals. Farmed Polish foxes are genetically more similar to the population of wild animals from North America than they are to the free-living population in Poland, as confirmed by the fact that the farmed animals are descended from animals raised in Canada.

We have developed a novel method for efficient structuring of the surface of materials by applying femtosecond near infrared laser pulses simultaneously with a weak extreme ultraviolet beam, which leads to a very strong radiation-matter interaction and brings a dramatic increase of the surface processing speed. We present our recent experimental results on surface nanostructuring of thin films of amorphous carbon and polymethyl methacrylate deposited on bulk substrates and discuss the underlying physical mechanisms. In the case of amorphous carbon, large areas of laser-induced periodic surface structures with a spatial period of 550 nm were created, having their origin in laser-induced convective currents. Our method provides a powerful tool for fast modification of tribological properties of the irradiated sample.

Assessment of Selenium Concentration in Selected Organs of Farmed Raccoon Dogs (Nyctereutes Procyonoides)The aim of the study was to determine selenium concentrations in the liver, kidneys, lungs, heart and muscles of farmed raccoon dogs (Nyctereutes procyonoides) and to evaluate their impact on hair coat quality. Selenium concentration was determined using the modified Watkinson's spectrofluorometric method. Subjects were 20 farmed raccoon dogs (Nyctereutes procyonoides) at the age of 8-9 months, which were kept on a farm in south-eastern Poland. The results show that liver selenium content averaged 0.23±0.10 μg/g w.w. (wet weight). The concentrations ranged from 0.04 to 0.49 μg/g w.w. Kidney selenium concentration (0.49±0.17 μg/g w.w. on average) was over twice that of liver concentration. Animals with higher scores for hair coat quality had lower selenium concentrations in the kidneys and liver, and higher selenium concentrations in muscles, but the differences were not significant. When relating Se concentrations determined in the liver of raccoon dogs to the biochemical criteria, it is concluded that 80% of the analysed raccoon dogs were deficient in this element and 20% had marginal levels. The results obtained in our study suggest that the food used on the farm did not fully meet the Se requirement of the raccoon dogs.

Asymmetric broadband connections in the home provide a limited upstream pipe to the Internet. This limitation makes various applications, such as remote backup and sharing high definition video, impractical. However, homes in a neighborhood often have high bandwidth wireless networks, whose bandwidth exceeds that of a single wired uplink. Moreover, most (wired and wireless) connections are idle most of the time. In this paper, we examine the fundamental requirements of a system that aggregates upstream broadband connections in a neighborhood using wireless communication between homes. A scheme addressing this problem must operate efficiently in an environment that is: i) highly lossy; ii) broadcast in nature; and iii) half-duplex. We propose a novel scheme, Link-alike, that addresses those three challenges using opportunistic wireless reception, a novel wireless broadcast rate control scheme, and preferential use of the wired downlink. Through analytical and experimental evaluation, we demonstrate that our approach provides significantly better throughput than previous solutions based on TCP or UDP unicast.

A multiplex PCR assay was developed for the detection and differentiation of the Yersinia enterocoliticaand Yersinia pseudotuberculosisisolates in both pure bacterial cultures and pig tonsils. The assay was based on the amplification of the ail, inv, yadA, and ystBgenes. The PCR products, corresponding to the ailgene and the plasmid-borne yadAgene or only one product corresponding to the ailgene, were detected in Y. enterocolitica4 biotype isolates. All of the Y. pseudotuberculosisisolates (n= 6) tested gave a positive PCR reaction for the invgene. For all tested Y. enterocolitica1A biotype isolates (n= 31), one product corresponding to the ystBgene was observed. The multiplex PCR assay was used to detect Y. enterocoliticaand Y. pseudotuberculosisstrains in pig tonsil samples obtained from 80 slaughtered pigs from three different herds. The presence of at least one of the specific PCR amplification products of ail-, ystB-, yadA-, and inv-specific sequences was observed in 11 samples (13.75%). These results of the multiplex PCR assay were compared with the results of conventional, microbiological testing. Y. enterocoliticaisolates were cultured from only 3 (3.75%) of the 80 pig tonsils examined. The multiplex PCR assay was shown to be an efficient tool for differentiation between the pYV plasmid–bearing Y. enterocoliticaisolates, the plasmidless Y. enterocoliticaisolates, the Y. enterocoliticabiotype 1A isolates, and the Y. pseudotuberculosisisolates with and without the pYV plasmid in naturally contaminated pig tonsils. This indicates that this assay is useful to control food processing and track the source of contamination.

ABSTRACTTo study the population structure of Enterococcus faecalisfrom Polish hospitals, 291 isolates were typed by pulsed-field gel electrophoresis and a novel multilocus sequence typing scheme (P. Ruiz-Garbajosa et al., J. Clin. Microbiol. 44:2220-2228, 2006). The isolates originated from geographically widespread medical institutions and were recovered during a 10-year period (1996 to 2005) from different clinical sources. The analysis grouped the isolates into five epidemic and 71 sporadic clones. The importance of the previously identified global clonal complexes CC2 and CC9 was corroborated by our findings that two of the Polish epidemic clones, A and J, were classified into these clonal complexes (CCs). However, the two most predominant clones, C (ST40) and F (CC87), did not cluster in the aforementioned CCs and may represent novel epidemic CCs. These clones may have emerged in Central Europe. Clone F, carrying glycopeptide resistance determinants of VanA or VanB phenotypes, caused several outbreaks in hematology units and appeared to be the most prevalent clone in recent years in Poland. Antimicrobial susceptibility testing and additional tests for pathogenicity-related phenotypes (hemolysin and gelatinase production) and genes (asa1and esp) were performed to further characterize these epidemic clones. Multidrug resistance, glycopeptide resistance, presence of asa1, and production of hemolysin appeared to be statistically significant features related to epidemicity. Production of gelatinase was significant for two of the epidemic clones, whereas presence of the espgene was not specific for the epidemic clones.

ABSTRACTA major limitation of adenovirus type 5 (Ad5)-based gene therapy, the inability to target therapeutic genes to selected cell types, is attributable to the natural tropism of the virus for the widely expressed coxsackievirus-adenovirus receptor (CAR) protein. Modifications of the Ad5 fiber knob domain have been shown to alter the tropism of the virus. We have developed a novel system to rapidly evaluate the function of modified fiber proteins in their most relevant context, the adenoviral capsid. This transient transfection/infection system combines transfection of cells with plasmids that express high levels of the modified fiber protein and infection with Ad5.ßgal.?F, an E1-, E3-, and fiber-deleted adenoviral vector encoding ß-galactosidase. We have used this system to test the adenoviral transduction efficiency mediated by a panel of fiber protein mutants that were proposed to influence CAR interaction. A series of amino acid modifications were incorporated via mutagenesis into the fiber expression plasmid, and the resulting fiber proteins were subsequently incorporated onto adenoviral particles. Mutations located in the fiber knob AB and CD loops demonstrated the greatest reduction in fiber-mediated gene transfer in HeLa cells. We also observed effects on transduction efficiency with mutations in the FG loop, indicating that the binding site may extend to the adjacent monomer in the fiber trimer and in the HI loop. These studies support the concept that modification of the fiber knob domain to diminish or ablate CAR interaction should result in a detargeted adenoviral vector that can be combined simultaneously with novel ligands for the development of a systemically administered, targeted adenoviral vector.

A growing number of antiangiogenesis strategies have been investigated for the treatment of cancer and other angiogenesis-dependent diseases. One of the most promising strategies is to systemically administer one or more antiangiogenic proteins frequently enough to achieve a sufficient long-term steady state level of the protein(s) to achieve the maximum beneficial effect. However, the utility of this strategy is limited because of many technical difficulties, including obtaining both the quantity and quality of the protein(s) necessary for optimal therapeutic benefit. To overcome these difficulties, we hypothesized that a single administration of a replication-defective adenoviral vector expressing a secretable antiangiogenic protein could achieve an optimal long-term systemic concentration. We constructed a recombinant adenoviral vector, Av3mEndo, which encodes a secretable form of murine endostatin. We demonstrated secretion of endostatin from several cell lines transduced with Av3mEndo. Partially purified endostatin secreted from Av3mEndo-transduced mammalian cells was shown to potently inhibit endothelial cell migration in vitro. A single intravenous administration of Av3mEndo in mice was shown to result in (1) prolonged and elevated levels of circulating endostatin, (2) partial inhibition of VEGF-induced angiogenesis in a VEGF implant angiogenesis model, and (3) prolonged survival and in 25% of mice the complete prevention of tumor growth in a prophylactic human colon/liver metastasis xenograft murine model. These results support our contention that adenoviral vectormediated expression of an antiangiogenic protein(s) represents an attractive therapeutic approach to cancer and other angiogenesis-dependent diseases.

ABSTRACTHepatocyte growth factor/scatter factor (HGF/SF) stimulates numerous cellular activities capable of contributing to the metastatic phenotype, including growth, motility, invasiveness, and morphogenetic transformation. When inappropriately expressed in vivo, an HGF/SF transgene induces numerous hyperplastic and neoplastic lesions. NK1 and NK2 are natural splice variants of HGF/SF; all interact with a common receptor, Met. Although both agonistic and antagonistic properties have been ascribed to each isoform in vitro, NK1 retains the full spectrum of HGF/SF-like activities when expressed as a transgene in vivo. Here we report that transgenic mice broadly expressing NK2 exhibit none of the phenotypes characteristic of HGF/SF or NK1 transgenic mice. Instead, when coexpressed in NK2-HGF/SF bitransgenic mice, NK2 antagonizes the pathological consequences of HGF/SF and discourages the subcutaneous growth of transplanted Met-containing melanoma cells. Remarkably, the metastatic efficiency of these same melanoma cells is dramatically enhanced in NK2 transgenic host mice relative to wild-type recipients, rivaling levels achieved in HGF/SF and NK1 transgenic hosts. Considered in conjunction with reports that in vitro NK2 induces scatter, but not other activities, these data strongly suggest that cellular motility is a critical determinant of metastasis. Moreover, our results demonstrate how alternatively structured ligands can be exploited in vivo to functionally dissociate Met-mediated activities and their downstream pathways.