Publications by authors named "Mirosława Ferens-Sieczkowska"

21 Publications

  • Page 1 of 1

Glycoproteins Presenting Galactose and -Acetylgalactosamine in Human Seminal Plasma as Potential Players Involved in Immune Modulation in the Fertilization Process.

Int J Mol Sci 2021 Jul 8;22(14). Epub 2021 Jul 8.

Department of Biochemistry and Immunochemistry, Wroclaw Medical University, 50-369 Wrocław, Poland.

In light of recent research, there is increasing evidence showing that extracellular semen components have a significant impact on the immune reaction of the female partner, leading to the tolerogenic response enabling the embryo development and implantation as well as further progress of healthy pregnancy. Seminal plasma glycoproteins are rich in the unique immunomodulatory glycoepitopes that may serve as ligands for endogenous lectins that decorate the surface of immune cells. Such interaction may be involved in modulation of the maternal immune response. Among immunomodulatory glycans, Lewis type antigens have been of interest for at least two decades, while the importance of T/Tn antigens and related structures is still far from understanding. In the current work, we applied two plant lectins capable of distinguishing glycoepitopes with terminal GalNAc and Gal to identify glycoproteins that are their efficient carriers. By means of lectin blotting and lectin affinity chromatography followed by LC-MS, we identified lactotransferrin, prolactin inducible protein as well as fibronectin and semenogelins 1 and 2 as lectin-reactive. Net-O-glycosylation analysis results indicated that the latter three may actually carry T and/or Tn antigens, while in the case of prolactin inducible protein and lactotransferrin LacdiNAc and lactosamine glycoepitopes were more probable. STRING bioinformatics analysis linked the identified glycoproteins in the close network, indicating their involvement in immune (partially innate) processes. Overall, our research revealed potential seminal plasma ligands for endogenous Gal/GalNAc specific lectins with a possible role in modulation of maternal immune response during fertilization.
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http://dx.doi.org/10.3390/ijms22147331DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8303229PMC
July 2021

Application of selected biosensor techniques in clinical diagnostics.

Expert Rev Mol Diagn 2021 Jul 31:1-13. Epub 2021 Jul 31.

Department of Chemistry and Immunochemistry, Medical University of Wroclaw, Wrocław, Poland.

Introduction: Examination of disease biomarkers mostly performed on crude materials, such as serum, meets some obstacles, resulting from sample complexity and the wide range of concentrations and sizes of the components. Techniques currently used in clinical diagnostics are usually time-consuming and expensive. The more sensitive and portable devices are needed for early diagnostics. Chemical sensors are devices that convert chemical information into parameters suitable for fast and precise processing and measurement.

Area Covered: We review the use of biosensors and their possible application in early diagnostics of some diseases like cancer or viral infections. We focus on different types of biorecognition and some technical modifications, lowering the limit of detection potentially attractive to medical practitioners.

Expert Opinion: Among the new diagnostic strategies, the use of biosensors is of increasing interest. In these techniques, the capture ligand interacts with the analyte of interest. Measuring interactions between partners in real time by surface plasmon resonance yields valuable information about kinetics and affinity in a short time and without labels. Importantly, the tendency in such techniques is to make biosensor devices smaller and the test results apparent with the naked eye, so they can be used in point-of-care medicine.
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http://dx.doi.org/10.1080/14737159.2021.1957833DOI Listing
July 2021

High-throughput glycopeptide profiling of prostate-specific antigen from seminal plasma by MALDI-MS.

Talanta 2021 Jan 13;222:121495. Epub 2020 Aug 13.

Leiden University Medical Center, Center for Proteomics and Metabolomics, 2300, RC, Leiden, the Netherlands. Electronic address:

An altered total seminal plasma glycosylation has been associated with male infertility, and the highly abundant seminal plasma glycoprotein prostate-specific antigen (PSA) plays an important role in fertilization. However, the exact role of PSA glycosylation in male fertility is not clear. To understand the involvement of PSA glycosylation in the fertilization process, analytical methods are required to study the glycosylation of PSA from seminal plasma with a high glycoform resolution and in a protein-specific manner. In this study, we developed a novel, high-throughput PSA glycopeptide workflow, based on matrix-assisted laser desorption/ionization-mass spectrometry, allowing the discrimination of sialic acid linkage isomers via the derivatization of glycopeptides. The method was successfully applied on a cohort consisting of seminal plasma from infertile and fertile men (N = 102). Forty-four glycopeptides were quantified in all samples, showing mainly complex-type glycans with high levels of fucosylation and sialylation. In addition, N,N-diacetyllactosamine (LacdiNAc) motives were found as well as hybrid-type and high mannose-type structures. Our method showed a high intra- and interday repeatability and revealed no difference in PSA glycosylation between fertile and infertile men. Next to seminal plasma, the method is also expected to be of use for studying PSA glycopeptides derived from other biofluids and/or in other disease contexts.
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http://dx.doi.org/10.1016/j.talanta.2020.121495DOI Listing
January 2021

TMPE Derived from Saffron Natural Monoterpene as Cytotoxic and Multidrug Resistance Reversing Agent in Colon Cancer Cells.

Int J Mol Sci 2020 Oct 13;21(20). Epub 2020 Oct 13.

Department of Chemistry and Immunochemistry, Wroclaw Medical University, ul. M. Skłodowskiej-Curie 48/50, 50-369 Wrocław, Poland.

Terpenes constitute one of the largest groups of natural products. They exhibit a wide range of biological activities including antioxidant, anticancer, and drug resistance modulating properties. Saffron extract and its terpene constituents have been demonstrated to be cytotoxic against various types of cancer cells, including breast, liver, lung, pancreatic, and colorectal cancer. In the present work, we have studied anticancer properties of TMPE, a newly synthesized monoterpene derivative of β-cyclocitral-the main volatile produced by the stigmas of unripe crocuses. TMPE presented selective cytotoxic activity to doxorubicin-resistant colon cancer cells and was identified to be an effective MDR modulator in doxorubicin-resistant cancer cells. Synergy between this derivative and doxorubicin was observed. Most probably, TMPE inhibited transport activity of ABCB1 protein without affecting its expression level. Analysis of TMPE physicochemical parameters suggested it was not likely to be transported by ABCB1. Molecular modeling showed TMPE being more reactive molecule than the parental compound-β-cyclocitral. Analysis of electrostatic potential maps of both compounds prompted us to hypothesize that reduced reactivity as well as susceptibility to electrophilic attack were related to the lower general toxicity of β-cyclocitral. All of the above pointed to TMPE as an interesting candidate molecule for MDR reversal in cancer cells.
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http://dx.doi.org/10.3390/ijms21207529DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7589963PMC
October 2020

The Potential Role of Seminal Plasma in the Fertilization Outcomes.

Biomed Res Int 2019 20;2019:5397804. Epub 2019 Aug 20.

Department of Chemistry and Immunochemistry, Wroclaw Medical University, Wroclaw, Poland.

For human infertility both male and female factors may be equally important. Searching for molecular biomarkers of male infertility, neglected for decades, and the attempts to explain regulatory mechanisms of fertilization become thus extremely important. Apart from examination of the structure and function of male gametes, also the possible importance of seminal plasma components should be considered. In this article we discuss data that indicate for the substantial significance of active seminal plasma components for conception and achievement of healthy pregnancy. Seminal plasma impact on the storage and cryopreservation of human and animal sperm and regulatory role of glycodelin on human sperm capacitation as well as hypothesized course of female immune response to allogenic sperm and conceptus has been discussed. The possible involvement of carbohydrates in molecular mechanism of fetoembryonic defense has been also mentioned.
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http://dx.doi.org/10.1155/2019/5397804DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6720062PMC
February 2020

Long-term changes of salivary exoglycosidases and their applicability as chronic alcohol-drinking and dependence markers.

World J Biol Psychiatry 2019 01 29;20(1):64-75. Epub 2017 Jun 29.

b Chemistry & Immunochemistry Dept , Medical University of Wrocław , Wrocław , Poland.

Objectives: Investigation of long-term dynamic changes of salivary activity/output of exoglycosidases, deglycosylation processes and their applicability as alcohol markers.

Methods: Exoglycosidase (α-fucosidase (FUC), β-galactosidase (GAL), β-glucuronidase (GLU), β-hexosaminidase (HEX, HEX A and HEX B isoenzymes) and α-mannosidase (MAN)) activities were measured in the saliva of healthy social drinking controls (C), alcohol-dependent non-smokers (ANS) and alcohol-dependent smokers (AS) at the 1st, 15th, 30th and 50th day of abstinence after chronic alcohol drinking.

Results: The activity of exoglycosidases was 2-3-fold (MAN), 2-6 fold (FUC), 8-25-fold (HEX A) and 19-40-fold (GLU) higher in the ANS and AS groups than in controls, and had good/excellent sensitivity, specificity and accuracy. The higher outputs of exoglycosidases were in the AS and ANS groups than in controls at the 1st day (GLU, HEX A) and at the 50th day (GLU, FUC, MAN) of abstinence. We found numerous correlations between alcohol-drinking days with GLU and HEX A, alcohol amounts with HEX A and duration of alcohol dependence with FUC and MAN activity/output.

Conclusions: Salivary exoglycosidases/deglycosylation processes were still very high up to 50 days after the end of alcohol consumption. We found markers of chronic alcohol consumption (HEX A), alcohol dependence (FUC and MAN) and chronic alcohol consumption and dependence (GLU).
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http://dx.doi.org/10.1080/15622975.2017.1337221DOI Listing
January 2019

Preliminary MALDI-TOF-MS analysis of seminal plasma N-glycome of infertile men.

Carbohydr Res 2016 Nov 18;435:19-25. Epub 2016 Sep 18.

Department of Chemistry and Immunochemistry, Wrocław Medical University, Bujwida 44A, 50-345 Wrocław, Poland. Electronic address:

Glycosylation pattern within reproductive tract is now suggested to be involved in providing female immune tolerance for allograft sperm and developing embryo, but the information whether impaired glycosylation may influence male fertility potential is still limited. We have analyzed seminal plasma N-glycome in pooled samples derived from fertile and infertile men by means of MALDI-TOF/TOF tandem mass spectrometry. Among infertile subjects, normozoospermic, oligozoospermic, asthenozoospermic and oligoasthenozoospermic samples were obtained. Eighty-six oligosaccharides were identified in all the analyzed samples. Differences in the content of unique glycans: high mannose and hybrid type, lacking terminal sialic acid and highly fucosylated were found when samples derived from infertile subjects with different semen patterns were compared to the fertile control. The content of highly branched glycans was 3-fold elevated in normozoospermic infertile men, while the expression of highly fucosylated oligosaccharides was increased in asthenozoospermic, oligozoospermic and oligoasthenozoospermic samples. Sialylation of oligosaccharides was decreased in oligozoospermic, oligoasthenozoospermic and especially asthenozoospermic samples, but increased in infertile normozoospermic subjects. Altered glycosylation observed in seminal plasma may reflect similar changes in sperm surface glycoproteins, and may disturb sperm interaction with female immune system. We suggest that at least some cases of unexplained male infertility may be associated with impaired glycosylation.
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http://dx.doi.org/10.1016/j.carres.2016.09.009DOI Listing
November 2016

Changes in glycosylation of human blood plasma chitotriosidase in patients with type 2 diabetes.

Glycoconj J 2016 Feb;33(1):29-39

Human blood plasma chitotriosidase (CHIT1) is a glycoprotein with chitinolytic activity with not fully elucidated biological function. Its increased level is observed in type 2 diabetes mellitus (T2DM) and is associated with development of diabetic complications. The CHIT1 glycosylation profile and degree is still poorly studied and never investigated in T2DM. Therefore the aim of the present study was to examine the association between glycosylation profile and degree and diabetes with accompanying nephropathy. In blood plasma of 28 patients with T2DM and 11 healthy subjects the CHIT1 concentration and specific activity were examined. The profile and degree of CHIT1 glycosylation were determined by lectin-ELISA using lectins specific to O-glycans (Jacalin, MPL, VVL) and sialo-specific SNA and MAA. We revealed that both concentration and specific activity of CHIT1 significantly increased in T2DM, especially in nephropathy with elevated albuminuria. The relative reactivities with lectins, except Jacalin, decreased progressively with T2DM occurrence and albuminuria progression. The most significant differences were observed between control vs. albuminuric group (Micro and Macro). It is also possible that the observed differences in immunoblotting pattern in molecular masses of CHIT1 bands between T2DM patients and healthy subjects may be caused by the differences in degree of CHIT1 glycosylation. The analysis of CHIT1 glycosylation status and the determination of CHIT1 concentration together with its enzymatic activity in blood plasma might constitute additional valuable diagnosis tools for the evaluation the T2DM patients with accompanying nephropathy. Extension of the lectin panel specific to O-glycans occurs useful for the further research using microarray formats, which are expected to accelerate “lectin-based glycan profiling” of glycoproteins.
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http://dx.doi.org/10.1007/s10719-015-9629-zDOI Listing
February 2016

Corrigendum to "The Analysis of Sialylation, N-Glycan Branching, and Expression of O-Glycans in Seminal Plasma of Infertile Men".

Dis Markers 2015 29;2015:652781. Epub 2015 Jul 29.

Department of Chemistry and Immunochemistry, Wrocław Medical University, Bujwida 44a, 50-345 Wrocław, Poland.

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http://dx.doi.org/10.1155/2015/652781DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4533124PMC
September 2015

Terminal Mannose Residues in Seminal Plasma Glycoproteins of Infertile Men Compared to Fertile Donors.

Int J Mol Sci 2015 Jul 2;16(7):14933-50. Epub 2015 Jul 2.

Department of Chemistry and Immunochemistry, Wrocław Medical University, Bujwida 44A, 50-345 Wrocław, Poland.

The impact of seminal plasma components on the fertilization outcomes in humans is still under question. The increasing number of couples facing problems with conception raises the need for predictive biomarkers. Detailed understanding of the molecular mechanisms accompanying fertilization remains another challenge. Carbohydrate-protein recognition may be of key importance in this complex field. In this study, we analyzed the unique glycosylation pattern of seminal plasma proteins, the display of high-mannose and hybrid-type oligosaccharides, by means of their reactivity with mannose-specific Galanthus nivalis lectin. Normozoospermic infertile subjects presented decreased amounts of lectin-reactive glycoepitopes compared to fertile donors and infertile patients with abnormal semen parameters. Glycoproteins containing unveiled mannose were isolated in affinity chromatography, and 17 glycoproteins were identified in liquid chromatography-tandem mass spectrometry with electrospray ionization. The N-glycome of the isolated glycoproteins was examined in matrix-assisted laser desorption ionization mass spectrometry. Eleven out of 27 identified oligosaccharides expressed terminal mannose residues, responsible for lectin binding. We suggest that lowered content of high-mannose and hybrid type glycans in normozoospermic infertile patients may be associated with impaired sperm protection from preterm capacitation and should be considered in the search for new infertility markers.
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http://dx.doi.org/10.3390/ijms160714933DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4519880PMC
July 2015

The analysis of sialylation, N-glycan branching, and expression of O-glycans in seminal plasma of infertile men.

Dis Markers 2015 29;2015:941871. Epub 2015 Mar 29.

Department of Chemistry and Immunochemistry, Wrocław Medical University, Bujwida 44a, 50-345 Wrocław, Poland.

Carbohydrates are known to mediate some events involved in successful fertilization. Although some studies on the glycosylation of seminal plasma proteins are available, the total glycan profile was rarely analyzed as a feature influencing fertilization potential. In this work we aimed to compare some glycosylation traits in seminal plasma glycoproteins of fertile and infertile men. The following findings emerge from our studies: (1) in human seminal plasma the presence and alterations of O-linked glycans were observed; (2) the expression of SNA-reactive sialic acid significantly differs between asthenozoospermia and both normozoospermic (fertile and infertile) groups; (3) the expression of PHA-L-reactive highly branched N-glycans was significantly lower in oligozoospermic patients than in both normozoospermic groups. Indication of the appropriate lectins that would enable the possibly precise determination of the glycan profile seems to be a good supplement to mass spectrum analysis. Extension of the lectin panel is useful for the further research.
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http://dx.doi.org/10.1155/2015/941871DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4393897PMC
December 2015

Gelatinases and their tissue inhibitors are associated with oxidative stress: a potential set of markers connected with male infertility.

Reprod Fertil Dev 2016 Jun;28(7):1029-1037

Department of Toxicology, Wroc?aw Medical University, Borowska 211, 50-556 Wroc?aw, Poland.

The expression and activity of matrix metalloproteinases (MMPs) may be regulated by oxidative stress in various pathophysiological processes; therefore, the aim of the present study was to analyse the associations between the expression of the gelatinases MMP-9 and MMP-2 and their tissue inhibitors TIMP-1, TIMP-2 and levels of total antioxidant capacity (TAC) and advanced oxidation protein products (AOPP) in seminal plasma prepared for artificial insemination. Levels of MMPs and TIMPs were evaluated using ELISA, whereas TAC and AOPP in the seminal plasma of 131 childless men and 38 fertile volunteers were determined spectrophotometrically. Seminal MMP-9 expression was higher in childless men than in fertile subjects, whereas there was no significant differences in MMP-2 expression between the analysed seminal groups. TIMP-1 and TIMP-2 expression was similar in all groups. However, TAC expression was significantly higher in infertile normozoospermic and oligozoospermic men and AOPP expression was higher in astheno-, oligo- and normozoospermic infertile patients than in fertile men. High AOPP, together with an increased MMP-9:TIMP-1 ratio alters the oxidative-antioxidative balance of the ejaculate, thereby reducing male fertility, and therefore these parameters may serve as additional diagnostic markers of semen quality and male reproductive potential.
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http://dx.doi.org/10.1071/RD14268DOI Listing
June 2016

Glycoprotein fucosylation is increased in seminal plasma of subfertile men.

Asian J Androl 2015 Mar-Apr;17(2):274-80

Department of Chemistry and Immunochemistry, Medical University of Wrocław, Wrocław, Poland.

Fucose, the monosaccharide frequent in N- and O-glycans, is a part of Lewis-type antigens that are known to mediate direct sperm binding to the zona pellucida. Such interaction was found to be inhibited in vitroby fucose-containing oligo- and polysaccharides, as well as neoglycoproteins. The objective of this study was to screen seminal plasma proteins of infertile/subfertile men for the content and density of fucosylated glycoepitopes, and compare them to samples of fertile normozoospermic subjects. Seminal proteins were separated in polyacrylamide gel electrophoresis and blotted onto nitrocellulose membrane and probed with fucose-specific Aleuria aurantia lectin (AAL). Twelve electrophoretic bands were selected for quantitative densitometric analysis. It was found that the content, and especially the density of fucosylated glycans, were higher in glycoproteins present in seminal plasma of subfertile men. No profound differences in fucosylation density were found among the groups of normozoospermic, oligozoospermic, asthenozoospermic, and oligoasthenozoospermic subfertile men. According to the antibody probing, AAL-reactive bands can be attributed to male reproductive tract glycoproteins, including prostate-specific antigen, prostatic acid phosphatase, glycodelin and chorionic gonadotropin. Fibronectin, α1 -acid glycoprotein, α1 -antitrypsin, immunoglobulin G and antithrombin III may also contribute to this high fucosylation. It is suggested that the abundant fucosylated glycans in the sperm environment could interfere with the sperm surface and disturb the normal course of the fertilization cascade.
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http://dx.doi.org/10.4103/1008-682X.138187DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4650452PMC
December 2015

Glycosylation changes in the salivary glycoproteins of alcohol-dependent patients: a pilot study.

Alcohol Alcohol 2014 Jan-Feb;49(1):23-30. Epub 2013 Oct 1.

Corresponding author: Chemistry and Immunochemistry Department, Medical University of Wrocław, Bujwida 44A, 51-345 Wrocław, Poland.

Aim: Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. However, total amount of sialic acid is not the only glycoepitope that may be altered as a result of the disease. This work is focused on glycan structures altered in salivary glycoproteins of alcoholics, indicating the most efficient carriers of such marker glycoepitopes.

Methods: Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans.

Results: In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. In lectin blotting ten glycoprotein bands were analyzed. The profile of disease-related alterations was found to be complex, but all six lectins studied here were able to detect altered glycan structures. In some glycoproteins the tendency to correct the glycosylation profile was observed after 7 weeks of abstinence.

Conclusion: Alterations in the glycosylation profiles in the salivary glycoproteins of alcohol-dependent people were found. Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. Further studies may allow one to estimate if such glycomarkers may also reflect the amount of alcohol intake or the duration of alcohol intake.
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http://dx.doi.org/10.1093/alcalc/agt152DOI Listing
August 2014

Changes in fucosylation of human seminal IgG and secretory component of IgA in leukocytospermic patients.

Glycoconj J 2014 Jan 22;31(1):51-60. Epub 2013 Sep 22.

Department of Chemistry and Immunochemistry, Wrocław Medical University, Bujwida 44a, 50-345, Wrocław, Poland,

Our study compares the status of human seminal plasma immunoglobulin G (IgG) and IgA secretory component (SC) fucosylation between infertile leukocytospermic and normal, fertile normozoospermic patients. The seminal IgG and SC are decorated with AAL-reactive core fucose, and antennary UEA- and LTA-reactive fucose of Lewis(y) and Lewis(x) structures, respectively. However, a correlation between IgG core fucosylation and IgG concentration (r = -0.52; p < 0.0003) was observed. The IgG present in leukocytospermic samples is characterized by lower expression of core fucose than in the normal group (0.82 ± 0.3 AU and 1.2 ± 0.3 AU, respectively; p < 0.002). In seminal plasma the SC is present in two forms: 78-kDa and 63-kDa. The present study has also shown a higher AAL and LTA specific reactivity of glycans expressed in 63-kDa SC, in comparison to 78-kDa SC, in the normal group. In leukocytospermia, the values of specific lectin reactivity for core fucose, fucose α(1-2)- and α(1-3)- linked, were similar for both SC bands. Moreover, the present study has shown that in leukocytospermic samples the mean concentrations of IgG and S-IgA are twice as high (131.68 ± 102.6 mg/l and 36 ± 27 mg/l, respectively) as in the normal group (67.68 ± 29.2 mg/l; p < 0.02, and 19 ± 18 mg/l, p < 0.019, respectively). The analysis of IgG and SC fucosylation status and the determination of IgG and S-IgA concentrations in seminal plasma might constitute a valuable diagnosis tools for the evaluation of male infertility associated with leukocytospermia with accompanying inflammation.
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http://dx.doi.org/10.1007/s10719-013-9501-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3892108PMC
January 2014

Comparison of haptoglobin and alpha₁-acid glycoprotein glycosylation in the sera of small cell and non-small cell lung cancer patients.

Postepy Hig Med Dosw (Online) 2013 Aug 8;67:828-36. Epub 2013 Aug 8.

Department of Chemistry and Immunochemistry, Wrocław Medical University, Wrocław, Poland.

Introduction: Cancer-related carbohydrate epitopes, which are regarded as potential diagnostic and prognostic biomarkers, are carried on the main acute phase proteins. It is not clear, however, if the glycosylation profile is similar in different glycoproteins, or it is protein specific to some extent. The aim of the study was to compare fucosylation, α2,3 sialylation and expression of sialyl-Lewisx epitopes (sLe(x)) in the serum as a whole, AGP and haptoglobin of small cell (SCLC) and non-small cell lung cancer (NSCLC) patients with respect to healthy subjects as well as the cancer stage and its histological type.

Material And Methods: Thirty-three NSCLC, 13 SCLC patients and 20 healthy volunteers were included in the study. Carbohydrate epitopes were detected by means of their reactivity with specific lectins and monoclonal anti-sLe(x) antibodies in direct or dual-ligand ELISA tests.

Results: Significantly increased fucosylation was found in total serum in both cancer groups and in NSCLC haptoglobin. No difference was observed in SCLC haptoglobin or α₁-acid glycoprotein in both cancer groups. Also α2,3 sialylation was elevated in total serum, but not in α₁-acid glycoprotein. This type of sialylation was undetectable in haptoglobin by means of MAA reactivity, in both healthy and cancer subjects. Complete sLe(x) antigens were overexpressed in total NSCLC serum and SCLC AGP, and their level was considerably lowered in cancer haptoglobin.

Discussion: Typical acute phase proteins, haptoglobin and AGP, exhibit different glycosylation profiles in lung cancer. Alterations observed in haptoglobin reflected the disease process better than those in AGP. Comparison of haptoglobin and AGP glycosylation to that observed in total serum suggests that some efficient carriers of disease-altered glycoproteins still remain unidentified.
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http://dx.doi.org/10.5604/17322693.1061788DOI Listing
August 2013

Seminal plasma glycoproteins in male infertility and prostate diseases: is there a chance for glyco-biomarkers?

Biomarkers 2013 Feb 15;18(1):10-22. Epub 2012 Oct 15.

Wrocław Medical University, Department of Chemistry and Immunochemistry, Wrocław, Poland.

Context: Disturbed protein-carbohydrate interactions may underlie the molecular mechanism of some diseases of the male reproductive tract, including infertility and prostate diseases.

Objective: To summarize the current knowledge on the glycosylation patterns of glycodelin-S, fibronectin, prostate-specific antigen, and α(1)-acid glycoprotein.

Results: Some rare glycoepitopes have been found in seminal plasma glycoproteins: high-mannose and polylactosamine-type glycans, and N-glycans containing N-acetyl-galactosamine. The glycosylation profiles occur altered in pathological conditions.

Conclusion: Further detailed studies may lead up to indicate the biomarkers useful in the management of male reproductive tract disorders.
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http://dx.doi.org/10.3109/1354750X.2012.719035DOI Listing
February 2013

Fucosylation of serum glycoproteins in lung cancer patients.

Clin Chem Lab Med 2005 ;43(4):361-9

Department of Chemistry and Immunochemistry, Wrocław Medical University, Poland.

Increased expression of sialyl Lewis X or A antigens on metastatic cancer cells leads to their selectin-mediated extravasation. Profound fucosylation of the serum microenvironment may be a factor that interrupts adhesion and influences the formation of metastases. In this study we quantitatively analyzed fucosylation of serum glycoproteins in small-cell and non-small-cell lung cancer patients. Fucosylation of four chosen glycoprotein bands was measured as the reactivity with Aleuria aurantia lectin on nitrocellulose blots, preceded by polyacrylamide gel electrophoresis. Relative fucosylation and fucosylation coefficients were calculated by densitometric analysis. Fucosylated oligosaccharides were observed in higher amounts in cancer sera when compared to sera from healthy individuals in all bands analyzed. Glycoproteins of a molecular mass of 29 kDa appear to carry more fucose residues than the 42-kDa band, comprising alpha(1)-acid glycoprotein and haptoglobin. Glycans of the 26-kDa band were fucosylated to a higher extent in non-small-cell vs. small-cell lung cancer. The results suggest that the extent of fucosylation could be a useful marker for estimation of the glycosylation status of serum proteins in cancer patients. Cluster analysis leads to the preliminary suggestion that the fucosylation status could serve as a predictive factor for patient survival.
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http://dx.doi.org/10.1515/CCLM.2005.066DOI Listing
September 2005

[Congenital disorders of glycosylation].

Postepy Hig Med Dosw 2003 ;57(4):425-44

Katedra i Zakład Chemii i Immunochemii Akademii Medycznej we Wrocławiu.

Congenital disorders of glycosylation are group of hereditary diseases resulting in severe psychomotor retardation and multiorgan failure. So far eleven different defects were identified on the pathway of N-glycan biosynthesis. Seven of them belong to CDG type I and result in incomplete occupation of potential N-glycosylation sites. Four defects were found in N-glycan processing. Molecular background of CDG and potential perspectives of therapy are summarised and updated in this review.
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April 2004

[The activity of N-acetyl-beta-hexosaminidase and gamma-glutamyltransferase in the serum of alcohol dependent people hospitalised after a long-lasting drinking period].

Psychiatr Pol 2003 May-Jun;37(3):495-502

Klinki Psychiatrii AM w Białymstoku.

Aim: The aim of the study was to compare the activity of N-acetyl-beta-hexosaminidase (HEX) and gamma-glutamyltransferase (GGT) in the serum of alcohol dependent people hospitalised after a long-lasting drinking period.

Method: The studied group consisted of 30 alcohol dependent men hospitalised after a long-lasting drinking period. The control group consisted of 30 healthy men--social drinkers. The activity of HEX was determined with colorimetric method Chatteriee (modified by Zwierz). The activity of GGT was determined by kinetic method.

Results: The medium activity of HEX in the serum in the studied group on the 1st day of abstinence was 2.3 times higher than the control group (p < 0.05). The activity of HEX decreased during abstinence. Starting from the 10th day of abstinence the activity of HEX was similar to the control group. The medium activity of GGT in the serum of the studied group on the 1st day of abstinence was 7 times higher than in the control group (p < 0.05). The activity of GGT decreased during abstinence. On the 30th day of abstinence the activity of GGT in the studied group was 2.7 times higher as compared to the control group (p < 0.05).

Conclusions: The activity of HEX and GGT in the serum significantly increases during long-lasting drinking period in alcohol dependent people. Increased HEX activity in the serum can be a more sensitive marker of long-lasting drinking period than GGT activity. The normalisation of HEX activity in the serum during abstinence in alcohol dependent people takes place significantly faster than the normalisation of GGT activity.
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December 2003

Glycoforms of six serum glycoproteins in a patient with congenital disorder of glycosylation type I.

Arch Immunol Ther Exp (Warsz) 2002 ;50(1):67-73

Department of Chemistry and Immunochemistry, Wrocław Medical University, Poland.

In this paper the occurrence and relative content of defectively glycosylated serum glycoforms in transferrin (Tf), alpha1-acid glycoprotein (AGP), haptoglobin (Hp), alpha1-antitrypsin (alpha1-AT), alpha2-macroglobulin (alpha2-MG) and ceruloplasmin (Cpl) in the serum of a patient with congenital disorder of glycosylation type I are reported. Blood samples were taken when the patient was 14 years old and then after a one-year interval. The patterns of glycoforms in both samples were compared. In 4 out of 6 examined glycoproteins, glycoforms lacking one and two oligosaccharide chains occurred. "Underglycosylated" glycoforms of alpha2-MG and Cpl were not clearly detectable. Tf was shown to be affected with this defect to a higher extent than other glycoproteins, containing only 30% properly glycosylated molecules and also as much as 30% of the molecules lacking two glycan units. In Hp and alpha1-AT the proportions of properly and defectively glycosylated forms were similar. This properly glycosylated form comprised 47% of the Hp and 51-55% of the alpha1-AT molecules. As in AGP and Tf, about 30% the of molecules lacked one glycan unit. Twenty-one percent of the Hp molecules were devoid of two glycans, and this amount slightly increased in the course of the year. In alpha1-AT, 19 and 17% of the molecules lacked two glycans in both samples, respectively. Only in AGP we did find a substantial difference between the two blood samples. In the course of the year, the amount of the form lacking 2-chains decreased from 12 to 3%, resulting in a simultaneous increase in the forms lacking one chain and the properly glycosylated. Our work also indicates, that applying a simple method of biochemical analysis such as SDS-PAGE/Western-blotting could be helpful in preliminary diagnosis and could improve the identification of congenital disorders of glycosylation.
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October 2002
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