Analyses of the Polymorphisms in E. coli Strains Associated with Heat-Shock Proteins Hsp 55 Isolated from Bird Feathers

The bird feathers are often colonized by pathogenic microorganisms including mainly bacteria of the E. coli species. There is a grooving evidence that due to colonization of the pathogenic bacteria after slaughter material may lead to different zoonosis diseases that endanger human health. Poultry diseases are a very important issue both economically and epidemiologically in each country. Currently, in practice, EU postmortem monitoring programs are often used to eliminate breeding poultry infected with different pathogenic microorganisms, e.g., E. coli by introducing mandatory bird vaccination. The article describes the combination of genetic and genomic methods that were used in the analysis of species specificity of strains and their genomes, including specific pathogenic bacteria in bird feathers. The aim of the study was (i) to investigate DNA polymorphisms of the obtained bacterial strains isolated from avian feathers (ii) obtaining recombinant Hsp55 protein and defining its role as a potential component of vaccines used in poultry diseases. For the detection and analysis of DNA polymorphisms, we have optimized a new innovative method based on the deficiencies of three molecular techniques, AFLP, PCR-MP, and PCR MP. This new method can be a useful tool used in the genotyping of bacterial E. coli serotypes present on avian feathers after the slaughter process. It also allows to effectively identify a number of early stages of infectious diseases from heterogeneous avian research material. Amplification of polymorphic regions was achieved by using a lower denaturation temperature of the primers and a reduction in the number of cycles in the classical PCR, which simplifies the procedure, preserving the quality and reproducibility of the obtained results. Research of recombinant Hsp55 protein has allowed us to determine the optimal conditions for its production by the classical methods used in proteomic analysis

POTENTIAL GENETIC AGENT BFL1 FOR TARGETED THERAPY IN CHRONIC LYMPHOCYTIC LEUKEMIA

Background: Many prognostic factors have been identified in chronic lymphocytic leukemia (CLL) but new ones are still desired. The biological characterization of CLL is now being translated into novel treatment strategies. One new prognostic factor, and therapeutic target, may be BFL1. It is both a serum and a molecular marker that contributes to the progression of CLL and its resistance to chemotherapy. The aim of this study was to evaluate the prognostic value of BFL1 and to assess its correlation with other known prognostic markers in CLL for the cladribine and cyclophosphamide regimen (CC). Methods: qPCR TaqMan® Low Density Array was used for gene expression measurements. Assessment of CD38, ZAP70 and BFL-1 proteins expression was done by means of flow cytometry. Serum TK activity was measured by immunoassay. Results: Protein BFL1 expression was found to be significantly higher in CLL patients than healthy volunteers (p=0.001). Moreover its level was significantly higher in patients with no response (NR) to CC therapy (p=0.009). The expression of BFL1 was considerably down regulated during CC treatment and BFL1 mRNA levels were inversely correlated with apoptotic response. In addition, protein BFL1 expression was found to be similar to thymidine kinase (TK) concentration regarding treatment response. As far as other markers are concerned, a positive correlation was identified between BFL1 and TK (r=0.52, p=0.01). Conclusions: Our findings suggest that BFL1 contributes to chemoresistance and may be a co-existing prognostic factor in CLL in the future

The sequence diversity and expression among genes of the folic acid biosynthesis pathway in industrial Saccharomyces strains

Folic acid is an important vitamin in human nutrition and its deficiency in pregnant women's diets results in neural tube defects and other neurological damage to the fetus. Additionally, DNA synthesis, cell division and intestinal absorption are inhibited in case of adults. Since this discovery, governments and health organizations worldwide have made recommendations concerning folic acid supplementation of food for women planning to become pregnant. In many countries this has led to the introduction of fortifications, where synthetic folic acid is added to flour. It is known that Saccharomyces strains (brewing and bakers' yeast) are one of the main producers of folic acid and they can be used as a natural source of this vitamin. Proper selection of the most efficient strains may enhance the folate content in bread, fermented vegetables, dairy products and beer by 100% and may be used in the food industry. The objective of this study was to select the optimal producing yeast strain by determining the differences in nucleotide sequences in the FOL2, FOL3 and DFR1 genes of folic acid biosynthesis pathway. The Multitemperature Single Strand Conformation Polymorphism (MSSCP) method and further nucleotide sequencing for selected strains were applied to indicate SNPs in selected gene fragments. The RT qPCR technique was also applied to examine relative expression of the FOL3 gene. Furthermore, this is the first time ever that industrial yeast strains were analysed regarding genes of the folic acid biosynthesis pathway. It was observed that a correlation exists between the folic acid amount produced by industrial yeast strains and changes in the nucleotide sequence of adequate genes. The most significant changes occur in the DFR1 gene, mostly in the first part, which causes major protein structure modifications in KKP 232, KKP 222 and KKP 277 strains. Our study shows that the large amount of SNP contributes to impairment of the selected enzymes and S. cerevisiae and S. pastorianus produce reduced amounts of the investigated metabolite. The results obtained here yield a list of genetically stable yeast strains which can be implemented as a starter culture in the food industry

Improving websites in terms of UX usability

Tematyka niniejszej pracy obejmuje nowy projekt i implementację, istniejącej już w sieci, strony internetowej fundacji charytatywnej, skupiając się na aspekcie użyteczności i estetyki witryny. W pracy omówione zostały podstawowe terminy związane z użytecznością oraz UX oraz uzasadniona została istotność wyżej wymienionych konceptów w budowaniu zaufania użytkowników do firmy lub organizacji. Praca uwzględnia analizę UX oraz UI poprzedniej wersji serwisu, uwydatniając, często niezauważalne na poziomie świadomym, błędy, mające wpływ na chęć korzystania z usług oferowanych przez właścicieli witryny. Zawarte są również badania z użytkownikami, mające na celu udowodnienie słuszności redesign’u oraz pokazanie, jakie emocje towarzyszą użytkownikom podczas korzystania z serwisów internetowych - zarówno tych niedbale i nieintuicyjnie zaprojektowanych, jak i tych spełniających normy estetyczne i funkcjonalne. Szczegółowa analiza wyników badań pokazuje skuteczność redesign’u i konieczność dbania o aspekty UX’owe podczas projektowania serwisów internetowych. Praca wzbogacona jest o porównania widoków starej oraz nowej wersji witryny oraz szczegółowe transkrypcje sesji testowych na obu wersjach serwisu.This thesis covers the redesign and reimplementation of an existing charity website, focusing on the usability and aesthetics aspects of the website. The paper discusses basic terms related to usability and UX and justifies the importance of these concepts in building user's trust in a company or organization. The paper includes a UX and UI analysis of the previous version of the website, highlighting the errors, often unnoticed on a conscious level, that affects the willingness to use the services offered by the owners of the website. It also includes user-centered research to prove the validity of the redesign and to show what emotions accompany users while using websites - both those carelessly and nonintuitively designed and those meeting aesthetic and functional standards. Detailed analysis of the research results shows the effectiveness of redesign and the necessity of taking care of UX aspects while designing websites. The paper is enriched with comparisons of views of the old and new versions of the website and detailed transcriptions of test sessions on both versions of the website

Research on the population of Grey wolf (Canis lupus) in Gorce National Park

Wilk (Canis lupus) jest gatunkiem szeroko rozpowszechnionym, prawie na całym świecie. Żyje w różnorodnych siedliskach od lasów strefy umiarkowanej po tereny pustynne. Wśród ludzi wzbudza zarówno pozytywne jak i negatywne skojarzenia. Przez wiele lat dążono do jego wytępienia i w wielu rejonach świata niestety odniosło to skutek. Wprowadzenie ochrony w kolejnych dekadach pozwoliło na zatrzymanie tego procesu likwidacji „niepożądanych” gatunków. Ustalenie zakresu zabiegów ochronnych jest ważnym zadaniem i wymaga stałego monitoringu tego gatunku. Stosuje się w tym celu między innymi analizę diety oraz analizę nagrań z fotopułapek. Metody te zostały również wykorzystane w niniejszej pracy. Badania prowadzone były na terenie Gorczańskiego Parku Narodowego. Dokonano analizy 166 odchodów wilka, na podstawie której wykazano, że najczęściej konsumowanym pokarmem były ssaki kopytne (95,23% biomasy), z przeważającą preferencją jelenia szlachetnego (Cervus elaphus). Uzupełnienie diety stanowiły średniej wielkości ssaki, takie jak bóbr (Castor fiber) czy borsuk (Meles meles) oraz materiał roślinny. Analiza 192 nagrań z udziałem tego gatunku potwierdziła występowanie wilków na terenie GPN, a najliczniej nagrana grupa liczyła 8 osobników. Wilki są najbardziej aktywne w okresie jesienno-zimowym, głównie nocą (68 ze 114 nagrań). Najczęściej przejawianym zachowaniem było przejście (58,5% wszystkich nagrań) oraz penetracja terenu (25,4%). Na nagraniach udało się zaobserwować także pogoń wilka z jeleniem.The wolf (Canis lupus) is a species with a wide geographic distribution worldwide. It inhabits a variety of habitats, from temperate forests to desert areas. Humans associate this species both with positive and negative connections. For many years it was exterminated and, in many cases, unfortunately this brought positive effect. However, the introduction of species protection allowed to stop this process. Establishing the type of protection procedures is an important task and requires constant monitoring of the species. Many methods are used for this purpose including diet analysis and use of camera traps. These two methods have been used in this work. The study was carried out in the Gorce National Park. An analysis of 166 wolf faeces showed that the most dominant food consumed were ungulates (95.23% BIO), with the prevailing preference of the red deer (Cervus elaphus). The diet was comprised by medium-sized mammals such as European beaver (Castor fiber) or European badger (Meles meles) and plant material. The analysis of 192 recordings from camera traps showed that the biggest group (pack) of wolves recorded in GPN included 8 individuals. Wolves are more active in autumn and winter seasons, and 68 out of 114 records with the presence of wolves were recorded at night. The most common manifested behaviour was the passing by (58.5%) and territorial penetration of the area (25.4% of all records). In one case the camera trap recorded pursuit of the red deer bull by the wolf

The role of research in UX (User Experience) design for computer applications

Tematyka niniejszej pracy obejmuje analizę tematu badań UX oraz ich roli w tworzeniu przydatnych i łatwych w użyciu produktów cyfrowych. W pracy omówione zostały podstawowe terminy związane z użytecznością, UX, oraz UCD, a także wyszczególnione zostały najważniejsze metody badawcze z zakresu pozyskiwania wymagań oraz ewaluacji użyteczności aplikacji komputerowych. Praca, oprócz szczegółowego omówienia poszczególnych metod zawiera także rozdziały dotyczące przygotowania do badań oraz analizy uzyskanych wyników. Zawarte są również transkrypcje i omówienie przeprowadzonych na potrzebę tej pracy badań z użytkownikami. Niniejsza praca zawiera zatem wiedzę teoretyczną dotyczącą tematu badań UX oraz ilustrację tej wiedzy na praktycznym przykładzie. Zamieszczone w niniejszej pracy informacje udowadniają słuszność i potrzebę przeprowadzania badań z użytkownikami, i ukazują jak cenne dla projektantów są informacje, które można dzięki nim uzyskać.This thesis covers the analysis of the topic of UX research and its role in creating useful and easy-to-use digital products. The paper discusses the basic terms related to usability, UX, and UCD, and details the most important research methods in the field of requirements capturing and usability evaluation of computer applications. In addition to a detailed overview of each method, the paper includes chapters on research preparation and analysis of the results. Also included are transcriptions and a review of the user research conducted for the purpose of this paper. Therefore, this paper contains theoretical knowledge on the topic of UX research and illustration of this knowledge with a practical example. The information included in this paper proves the validity and necessity of user research, and shows how valuable for designers are the information that can be obtained through it

Intra-strains diversity of expression of polymorphic PKS4 gene in comparison in zearalenone production by Fusarium graminearum during in vitro cultivation

Filamentous fungi belonging to the Fusarium genus are responsible for large economic losses due to their high pathogenicity and toxigenicity. Fusarium sp. may produce variety of mycotoxins, one of them is zearalenone (ZEA). The presence of the PKS4 gene shows the possibility of zearalenone biosynthesis by Fusarium sp. In this study, in four Fusarium graminearum and one Fusarium poae strains the presence of PKS4 genes and ZEA concentrations were determined. The presence of the PKS4 gene was confirmed by classical polymerase chain reaction (PCR) in three of four strains of F. graminearum. One strain with no PKS4 gene detected was found while still producing ZEA. In the present study, a real-time PCR assay has been successfully performed for the relative expression of Fusarium strains based on new designed primers targeting the PKS4 gene involved in ZEA biosynthesis. Result shows that P56/4 strain of F. graminearum has the highest mRNA level, in the range of 12, what correlates to the high production of this mycotoxin. In this study, a real-time PCR assay has been successfully developed for the prediction of the production of ZEA by F. graminearum strains by PCR real-time techniques based on primers targeting the gene, PKS4, involved in ZEA biosynthesis. The special significance was pointed to occurring genes polymorphism