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M Ugorski

Publications and source records attributed to M Ugorski.

At least 19 recordsLinked to original sources
PubMed2006

Differential effect of IFN-tau on proliferation and distribution of lymphocyte subsets in one-way mixed lymphocyte reaction in cows and heifers.

IFN-tau is a signaling protein secreted by the bovine conceptus during the peri-implantation period and responsible for pregnancy recognition. Its main role is the prevention of pulsatile release of luteolytic PGF2alpha, but it also exerts immunomodulatory activities characteristic for other type I interferons. The aim of the study was to examine the effect of IFN-tau on the proliferation and distribution of peripheral blood lymphocyte subsets during one-way mixed lymphocyte reaction (MLR) in cows and heifers. IFN-tau inhibited the proliferative response of lymphocytes in MLR both in cows and heifers in a dose-dependent manner, but cow lymphocytes were less susceptible than those ones from heifers. It was also showed that IFN-tau differentially changed lymphocyte subsets distribution in MLR in cows and heifers. In cows, the relative percentage of CD8(+) cells after MRL in the presence of IFN-tau was significantly lower than in heifers. Differential effect of rIFN-tau on proliferation and lymphocyte subsets distribution in a one-way MRL in cows and heifers indicated that the age of the mother is an important factor in immunomodulatory effect towards developing bovine embryo.

Analysis of Variance↗
PubMed2006

Analysis of the presence of ail, ystA and ystB genes in Yersinia enterocolitica strains isolated from aborting sows and aborted fetuses.

Forty-five Yersinia enterocolitica strains isolated from aborted fetuses and placentas and from vaginal and rectal swabs of aborting sows were subjected to serotyping, biochemical typing and polymerase chain reaction multiplex analyses to detect the presence of the ail, yst A and ystB genes. The isolates were recovered from the internal organs (tonsil, lung, liver, spleen, kidney, mesentheric lymph nodes, small intestine and rectal intestine) of 18 (18.6%) of 97 aborted fetuses examined, two (8%) of 25 aborted placentas and 27 (15.8%) of 172 examined aborting sows. Serotyping of Y. enterocolitica revealed that only six (13.3%) of the examined isolates belonged to serotype O:3, with a considerable number of isolates (31.1%) having serotype O:5, while biochemical studies showed that as many as 40 of the 45 strains belonged to biotype 1A. As expected, the Y. enterocolitica strains of bioserotype 4/O:3 contained ail and ystA genes, while strains of biotype 1A contained only the ystB gene.

Aborted Fetus↗
PubMed2005

Combined effect of surface electrostatic charge and poly(ethyl glycol) on the association of liposomes with colon carcinoma cells.

Particulate drug formulations are considered to be a means that may improve the pharmacokinetics and biodistribution of active compounds. By using them, drug distribution is determined solely by the properties of the carrier. The surface properties of such supramolecular aggregates determine how they will interact with various biological structures. Among others, surface electrostatic charge and surface grafted polymers are considered to be among the major factors affecting its interaction with proteins and cells. In this article, we present experimental evidence that properly selected surface electrostatic charge and grafted polymers can alter the association of liposomes with colon cancer cells. The dependence of the adsorption of liposomes onto the cell surface on the quantity and length of surface grafted polymers for a certain surface charge density exhibits a distinct maximum. For example, when liposomes were formed with 20 mol% of DOTAP, PE-PEG350 increased liposome adsorption by up to 6 mol%. This adsorption maximum depends on both polymer length and charge type. Results presented in this article show that the interaction of liposomes with colon cancer cells can be tuned by a proper combination of liposome surface electrostatics and surface grafted polymers.

Cell Line, Tumor↗
PubMed2005

Differentiation of Salmonella Gallinarum biovar Gallinarum from Salmonella Gallinarum biovar Pullorum by PCR-RFLP of the fimH gene.

In our studies on FimH adhesins expressed by different Salmonella serovars, we cloned and sequenced the fimH genes from Salmonella enterica ssp. Enterica ser. Gallinarum biovar Gallinarum and S. enterica ssp. Enterica ser. Gallinarum biovar Pullorum. Comparison of the nucleotide sequences revealed the presence of a single-nucleotide polymorphism (SNP) at position 544 bp from the A of the start codon of the fimH open reading frame (ORF). Further analysis of the restriction enzyme sites in fimH gene showed that the SNP at this position is responsible for a sequence specifically recognized by SacI in S. Gallinarum biovar Gallinarum only, making it possible to differentiate both biovars with the use of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Digestion of PCR amplicons of the fimH gene from S. Gallinarum biovar Gallinarum strains with SacI gave two DNA fragments of 554 and 472 bp and only one fragment of 1026 bp for S. Gallinarum biovar Pullorum. This allows a clear differentiation between these two biovars.

Animals↗
PubMed2005

Genotype analysis of Campylobacter jejuni isolated from broilers in Poland.

Twenty-six Campylobacter jejuni strains isolated from poultry were analyzed by genotypic typing including ITS-profiling, REP- and ERIC-PCR. ITS-profiling revealed the presence of 8 different genotypes. Amplification of REP sequences by PCR gave similar results with 10 different genotypes. ERIC-PCR was found to be the most discriminatory for typing C. jejuni. As many as 13 different DNA patterns were obtained with this technique. Based on data obtained it was found that C. jejuni isolates recovered from broilers at the slaughterhouses in southwest Poland are characterized by a high degree of genetic heterogeneity.

Animals↗
PubMed2004

Complex nature of the human antisperm antibody response in SCID mice.

Human peripheral blood mononuclear (PBMs) cells were introduced into the peritoneal cavity of severely-combined immunodeficient (SCID) mice in concentrations of 2.5-4.0 x 10(7) cells per mouse. Whole mononuclear cell suspensions were used either unstimulated or following primary in vitro culture with human spermatozoa. In some experiments, immunodepletion of CD8(+) cells was carried out prior to grafting. Lymphocytes were obtained from nonsensitized (to antigen) human subjects or from individuals who were primed in vivo (vasectomized individuals in case of sperm antigens). An enzyme-linked immunosorbent assay was employed to assess total human immunoglobulin (G or M) levels as well as the specificity of the antibodies generated. We have been successful by generating primary and secondary immune responses with 'naïve' human lymphocytes, challenged with chlamydia or ovalbumin but without adjuvant or CD8(+) immunodepletion; however, we were unable to induce specific antibodies to spermatozoa under this regime in SCID male mice. We then employed female SCID mice, treated with sperm antigen extracts (glycosylated or deglycosylated) encapsulated in liposomes and human lymphocytes obtained from 'naïve' or pre-sensitized in vivo subjects. It was found that the most pronounced humoral response to sperm antigens was obtained with deglycosylated antigens and PBMs from vasectomized (in vivo pre-primed to spermatozoa) individuals. A presented SCID mice model can be helpful at understanding of antisperm antibody development and the molecular nature of generated antibodies to modified sperm antigenic entities.

Adult↗
PubMed2004

Typing of Yersinia Enterocolitica Isolates by ITS profiling, REP- and ERIC-PCR.

Thirty-five Yersinia enterocolitica strains isolated from humans, pigs and foxes were analysed by genotyping including intergenic transcribed sequence (ITS) profiling, REP- and ERIC-PCR. ERIC-PCR revealed the presence of seven different genotypes. Amplification of the 16S-23S rDNA spacer region by ITS profiling gave similar results with nine different genotypes. REP-PCR was found to be more discriminatory for typing of Y. enterocolitica than ERIC-PCR and ITS profiling. Fifteen different DNA patterns were obtained by this technique. Based on data obtained by three methods it was found that: (i) Y. enterocolitica strains belonging to the same serotype can represent different genotypes and vice versa; (ii) isolates recovered from humans, pigs and foxes exhibit limited heterogeneity and, independent of the origin, one or two prevailing genotypes were always observed; and (iii) many human Y. enterocolitica isolates shared common genotypes with porcine isolates.

Animals↗
PubMed2002

Comparison of ITS profiling, REP- and ERIC-PCR of Salmonella Enteritidis isolates from Poland.

Thirty-one Salmonella Enteritidis strains isolated from chickens, broilers and hens were analysed by genotypic typing including REP-PCR. ERIC-PCR and ITS profiling (PCR-ribotyping). Analysis of DNA banding patterns generated by REP-PCR revealed the presence of 22 different genotypes, which were grouped by dendrogram analysis into three distinct lineages (maximum similarity approx. 50%). Each isolate of S. Enteritidis analysed by ERIC-PCR generated an individual DNA pattern. Again, these isolates could be divided into three distinct genomic groups (maximum similarity approx. 60%) by their ERIC-PCR fingerprints. REP- and ERIC-PCR were found to be more discriminatory for typing of S. Enteritidis than ITS profiling. Amplification of the 16S-23S rDNA spacer region gave nine different profiles of DNA, subdivided into two closely related groups by dendrogram analysis. In summary, data obtained by genotyping methods for S. Enteritidis isolates from regions located in the south-west and the central parts of Poland revealed an enormous heterogeneity among analysed samples, and proved that REP- and ERIC-PCR are highly discriminatory techniques, which can be used, in addition to conventional methods, in epidemiological studies of S. Enteritidis infections.

Animals↗
PubMed2001

Sialosyl Le(a)-carrying gangliosides present on the surface of colon carcinoma cells are not directly involved in adhesion to E-selectin.

We have shown previously that human colon cancer CX-1 cells contain lipid- and protein-bound sialosyl Lewis(a) structures that support the adhesion of these cells to E-selectin. Treatment of cancer cells with O-sialoglycoprotease did not decrease either the binding of anti-sialosyl Le(a) antibodies or binding to E-selectin-expressing CHO cells. This suggested that cleavage of sialomucins uncovered cryptic sialosyl Le(a) gangliosides that support such interactions. In the present study, inhibitors of glycolipid and O-glycan biosynthesis, d,l-threo-PPPP and GalNAc-alpha-O-benzyl, respectively, were used to study whether the binding of anti-sialosyl Le(a) antibody and adhesion of CX-1 cells to E-selectin can be mediated by sialosyl Le(a) gangliosides. Treatment of cancer cells with each of the inhibitors decreased the expression of the respective glycoconjugates as shown by TLC-binding assay and immunoblotting with anti-sialosyl Le(a) antibody. However, only slight differences in binding of antisialosyl Le(a) antibody to the surfaces of control and inhibitor-treated CX-1 cells were found by flow cytometry, as well no differences were observed in binding of control and inhibitor-treated CX-1 cells to E-selectin-expressing CHO cells, supporting the earlier hypothesis on the involvement of gangliosides in binding of anti-sialosyl Lewis(a) in the partial absence of mucin O-glycans. This hypothesis was further proven by electron microscopy data. Both native CX-1 and d,l-threo-PPPP-treated cells were labelled with anti-sialosyl Lewis(a) antibody mostly at a distance 70-90 nm from cell surface, suggesting interaction with protein-bound carbohydrate structures only. In contrast, the cancer cells treated with GalNAc-alpha-O-benzyl showed most of the staining around 20 nm distance from the plasmalemma, implying that the antibody interacts with lipid-bound sialosyl Lewis(a) instead. The electron microscopy data in conjunction with other results described in this report strongly support the hypothesis that sialosyl Lea gangliosides are not involved in the adhesion of CX-1 cells to E-selectin when mucins are present on the cell surface, but they may be involved in binding to E-selectin in their absence.

Animals↗
PubMed2000

[REP and ERIC repetitive DNA sequences in bacteria--diagnostic significance].

Main part of eukaryotic genomes is build of unique sequences coding proteins and RNAs, but they contain as well numerous repeats interspersed with single-copy fragments. Existence of repetitive sequences were also demonstrated in prokaryotic genomes. They are found in different species of Gram-negative and Gram-positive bacteria. Interspersed repetitive sequence elements called REP and ERIC sequences are present in different species of Enterobacteriaceae family, including Escherichia coli and Salmonella typhimurium. Their functions are not completely clear, probably they play important role in regulation of gene expression. Nevertheless, REP and ERIC elements are widely use in identification and genetic analysis of bacteria. For example, using rep-PCR technique it is possible to discriminate between closely related serovars of the same species, which enables to analyze phylogenetic and epidemiological relations among them.

Animals↗
PubMed2000

Study on ganglioside composition in brain tumours supra- and infratentorial.

The composition of gangliosides in primary tumors depends on their histological origin and differentiation grade (biological malignancy). The aim of the present study was the comparison of gangliosides in various types of brain tumours. The studies were performed on specimens from 20 high grade gliomas, 5 low grade gliomas, 8 meningiomas and 9 metastatic tumours. The isolated gangliosides were separated on silica 60 HPTLC plates and their mobilities were compared with glycolipids standards. The following observations were made: 1. high and low grade gliomas had similar ganglioside profiles comprising 7 different species; 2. the profiles of gangliosides isolated from metastatic neoplasms differ considerably from those of gliomas as well as menigiomas.

Adult↗
PubMed1999

CEA-related proteins on human urothelial cell lines of different transformation grades.

CEA family proteins from human urothelial cell lines of different transformation grades were characterized by flow cytometry and Western blotting using monoclonal antibodies: 26/3/13, D14HD11, 9A6 and 4/3/17. The following observations were made: (i) the urothelial cell lines, representing transformation grade III (TGr III, tumorigenic, invasive cells), were characterized by the presence of a component with molecular mass 110-135 kDa, most probably representing biliary glycoprotein (BGP); (ii) BGP was absent in non-tumorigenic and non-invasive TGr II urothelial cell lines; (iii) a protein band with apparent molecular mass 180 kDa, and migrating as a CEA standard was detected in only one of seven urothelial cell lines analyzed; (iv) a broad band of apparent molecular mass migrating at 65-90 kDa, probably representing NCA-50/90, was found in two tumorigenic and invasive cell lines, HCV 29T and Hu 1703He.

Carcinoembryonic Antigen↗
PubMed1998

Role of sialosyl Lewis(a) in adhesion of colon cancer cells--the antisense RNA approach.

To study whether the adhesion of colon cancer cells to E-selectin can be directly affected by changes in the expression level of sialosyl Le(a) antigen we created a specific loss-of-function phenotype. A stable subclone (CX-1.1) with high expression of sialosyl Le(a) structure, obtained from a heterogenous population of colon carcinoma CX-1 cells, was transfected with an expression vector containing a fragment of cDNA for alpha1,3/4-fucosyltransferase in antisense orientation. After transfection, the cell line was isolated which did not express sialosyl Le(a) antigen and lacked the alpha1,3/4-fucosyltransferase activity, despite an unchanged level of mRNA specific for this enzyme. It was found that the specific lack of expression of sialosyl Le(a) carbohydrate structure on the surface of colon cancer cells completely abolished their adhesion to E-selectin. To evaluate which cellular glycoconjugates carry sialosyl Le(a) antigen, glycoproteins as well as glycolipids of CX-1.1 cells were analysed for the expression of this structure. Anti-sialosyl Le(a) antibodies detected multiple glycoprotein bands with apparent molecular masses of 65-280 kDa on western blots, and an intense band representing sialosyl Le(a)-ganglioside on a thin-layer chromatogram. Using O-sialoglycoprotease from Pasteurella haemolytica and an alkaline beta-elimination procedure, it was shown that protein-linked sialosyl Le(a) structures are carried mostly by mucin-type glycoproteins. However, treatment of CX-1.1 cells with O-sialoglycoprotease did not decrease either their binding to E-selectin-expressing Chinese hamster ovary cells, or binding of anti-sialosyl Le(a) antibodies to the cell surface. These results suggested that cleavage of sialomucins uncovered cryptic sialosyl Le(a)-ganglioside, which was inaccessible for the antibody and E-selectin in untreated cells. This hypothesis was confirmed to some extent by the higher accessibility of gangliosides to galactose oxidase on the surface of O-sialoglycoprotease-treated CX-1.1 cells, comparing to untreated cells. We propose that glycoproteins as well as gangliosides carrying sialosyl Le(a) structures, when properly exposed and present in high density on surface of cancer cells, can effectively support the adhesion of cancer cells to E-selectin.

Animals↗
PubMed1998

Metastatic potential of human CX-1 colon adenocarcinoma cells is dependent on the expression of sialosyl Le(a) antigen.

Several lines of evidence indicate that sialosyl Le(a), tumor-associated carbohydrate antigen present on human colon carcinoma cells, is involved in formation of metastases. To study the role of this carbohydrate structure in development of metastases, we have used the clone of human colon carcinoma CX-1 cells transfected with antisense expression vector containing fragment of cDNA for alpha1,3/4-fucosyltransferase (FT III), which is involved in synthesis of sialosyl Le(a) tetrasaccharide. It has been reported previously that, in contrast to the parental cells, the antisense-transfected CX-1.1AS5 cells do not express sialosyl Le(a) and do not adhere to E-selectin-expressing CHO cells. In the present work we have studied the formation of liver metastases by CX-1.1AS5 cells after their orthotopic or intrasplenic implantation into athymic nu/nu mice. After orthotopic implantation of sialosyl Le(a)-negative colon carcinoma CX-1.1AS5 cells, the number of mice with liver metastases was markedly lower (21% of mice) in comparison with their number after implantation of the parental CX-1.1 cells (86% of mice). However, no differences in ability to form colonies in liver were observed between parental CX-1.1 cells and antisense-transfected CX-1.1AS5 cells after intrasplenic inoculation. The liver metastases were formed in 89% and 84% of mice, respectively. Our data support the thesis on the importance of sialosyl Le(a) antigen expression in the development of liver metastases by colon cancer cells, and indicate the role of transplantation route and primary tumor localization in formation of metastases.

Adenocarcinoma↗
PubMed1997

beta-Elimination of O-glycans from glycoproteins transferred to immobilon P membranes: method and some applications.

Selective beta-elimination of O-glycans from glycoproteins transferred from electrophoretic gels onto Immobilon P membranes is described. The experiments were performed with erythrocyte membrane proteins, in which glycophorins are the major poly-O-glycosylated components, and with lysates of human colon cancer cells CX-1.1. Lectins and monoclonal antibodies against peptidic, glycopeptidic, and carbohydrate epitopes were used to examine the effect of degradation. Experiments with erythrocyte membrane proteins showed that after heating the blots in 0.055 M NaOH for 16 h at 40 degrees C the O-glycans of glycophorins were undetectable, while N-glycans and peptidic epitopes of proteins were detected with unchanged or even increased intensity compared to untreated blots. The method was used to show that most protein-linked sialyl-Lea epitopes present on CX-1.1 cancer cells are located on O-glycosidic chains. Moreover, beta-elimination on the blots allows examination of the dependence of peptidic epitopes on O-glycosylation. This was shown using monoclonal antibodies specific for blood group M- or N-related epitopes of glycophorin A (GPA). Most of these antibodies recognize glycopeptidic epitopes dependent on O-glycosylation and, therefore, they did not detect GPA on NaOH-treated blots. Some less frequent anti-M antibodies cross-reacting with the rare GPA variant of Mg type are specific for a peptidic epitope which is unrelated to the MN blood group-specific amino acid sequence in unglycosylated peptides, but is recognized in GPA-M only in the glycosylated antigen. These antibodies, which showed specificity for GPA-M on untreated blots, detected GPA-M, GPA-N, and glycophorin B on NaOH-treated blots.

Antibodies, Monoclonal↗
PubMed1996

Adhesion of human uroepithelial cells to E-selectin: possible involvement of sialosyl LewisA-ganglioside.

In a previous study we showed that tumorigenic and invasive human uroepithelial cell lines are characterized by the presence of sialosyl Le(a) (sLe(a)) ganglioside. Our data suggested that expression of this glycolipid correlated with acquisition of the malignant phenotype by human urothelial cells. To evaluate the postulated adhesion function of sLe(a) antigen, we studied the adherence of 6 human urothelial cell lines with different expressions of this carbohydrate structure to E-selectin-expressing CHO cells. The only cell line that bound specifically to E-selectin was Hu 1703He, which expressed the highest level of sLe(a) antigen. The involvement of carbohydrate-E-selectin interaction in the adhesion of Hu 1703He cells was indicated by the following facts: (i) anti-E-selectin monoclonal antibody (MAb) completely abolished binding to E-selectin-expressing CHO cells; (ii) removal of sialic acid from Hu 1703He cells highly decreased the adhesion. Adhesion correlated with the presence of several sLe(a)-carrying glycoproteins, which was shown by immunoblotting of Hu 1703He cell lysate with anti-sLe(a) MAb 19-9. The binding of antibody was abolished when cell lysate was treated with O-sialoglycoprotein endopeptidase, suggesting that sLe(a) is present on O-linked oligosaccharides. However, incubation of Hu 1703He cells with O-sialoglycoprotease had no effect on adhesion to E-selectin or on binding of 19-9 MAb to the cell surface. Our data suggest that (i) protein-bound sLe(a) oligosaccharides represent only a minor portion of whole sLe(a) antigen produced by uroepithelial cells; (ii) effective binding to E-selectin occurs when sLe(a) oligosaccharide present on cell-surface glycosphingolipids is expressed in high density since the cell lines with moderate expression of sLe(a) ganglioside did not bind to E-selectin-transfected CHO cells.

Animals↗