This page has only limited features, please log in for full access.
Introduction: Protein profiling allows the determination of the presence of proteins marking various stages of the disease, and differentiates between people at risk of various diseases. In type 1 diabetes, protein profiling had been previously used to find blood markers other than islet autoantibodies to indicate the pancreatic beta cell destruction process and to reflect the progression of type 1 diabetes mellitus (T1DM). However, T1DM is an auto-immune disease and its clinical presentation changes in time of its duration. The aim of the study: To find differences in protein profiles in patients with type 1 diabetes according to diabetes control (HbA1c > 7%) and with presence of diabetic complications or obesity. It may help to identify subgroups of patients who may need a better clinical supervision and individualized treatment. Material and methods: A group of 103 patients with auto-immunologically confirmed T1DM, and meeting the following inclusion criteria: Caucasian race, duration of diabetes >5 years, were used in the study. Criteria of exclusion: past or present cancer (treated with chemo-/radiotherapy), diseases of the liver (ALT > 3 × ULN) except for people with simple hepatic steatosis, chronic renal disease (eGFR < 30 mL/1.73 m2/min), and acute inflammation (CRP > 5 mg/dL). The study group was divided in terms of the presence of chronic complications, obesity, or poor metabolic control (HbA1c > 7%). Protein profiling was completed by using the MALDI-TOF MS (matrix-assisted laser desorption/ionization-time of flight mass spectrometry) analyzer. Results: Differentiating proteins were identified in all of the groups. The groups burdened with complications, obesity, and poor metabolic control were characterized by increased levels of fibrinogen, complement C4 and C3. Conclusion: The groups of type 1 diabetes patients burdened with complications, obesity, and poor metabolic control were characterized by increased levels of fibrinogen, complement C4 and C3. Further detailed studies are necessary to determine more subtle changes in the proteomic profile of patients with type 1 diabetes.
Agnieszka Zawada; Dariusz Naskręt; Eliza Matuszewska; Zenon Kokot; Marian Grzymisławski; Dorota Zozulińska-Ziółkiewicz; Agnieszka Dobrowolska; Jan Matysiak. MALDI-TOF Protein Profiling Reflects Changes in Type 1 Diabetes Patients Depending on the Increased Amount of Adipose Tissue, Poor Control of Diabetes and the Presence of Chronic Complications. International Journal of Environmental Research and Public Health 2021, 18, 2263 .
AMA StyleAgnieszka Zawada, Dariusz Naskręt, Eliza Matuszewska, Zenon Kokot, Marian Grzymisławski, Dorota Zozulińska-Ziółkiewicz, Agnieszka Dobrowolska, Jan Matysiak. MALDI-TOF Protein Profiling Reflects Changes in Type 1 Diabetes Patients Depending on the Increased Amount of Adipose Tissue, Poor Control of Diabetes and the Presence of Chronic Complications. International Journal of Environmental Research and Public Health. 2021; 18 (5):2263.
Chicago/Turabian StyleAgnieszka Zawada; Dariusz Naskręt; Eliza Matuszewska; Zenon Kokot; Marian Grzymisławski; Dorota Zozulińska-Ziółkiewicz; Agnieszka Dobrowolska; Jan Matysiak. 2021. "MALDI-TOF Protein Profiling Reflects Changes in Type 1 Diabetes Patients Depending on the Increased Amount of Adipose Tissue, Poor Control of Diabetes and the Presence of Chronic Complications." International Journal of Environmental Research and Public Health 18, no. 5: 2263.
A growing interest in metabolomics studies of cultured cells requires development not only untargeted methods capable of fingerprinting the complete metabolite profile but also targeted methods enabling the precise and accurate determination of a selected group of metabolites. Proline metabolism affects many crucial processes at the cellular level, including collagen biosynthesis, redox balance, energetic processes as well as intracellular signaling. The study aimed to develop a robust and easy-to-use targeted metabolomics method for the determination of the intracellular level of proline and the other two amino acids closely related to proline metabolism: glutamic acid and arginine. The method employs hydrophilic interaction liquid chromatography followed by high-resolution, accurate-mass mass spectrometry for reliable detection and quantification of the target metabolites in cell lysates. The sample preparation consisted of quenching by the addition of ice-cold methanol and subsequent cell scraping into a quenching solution. The method validation showed acceptable linearity (r > 0.995), precision (%RSD < 15%), and accuracy (88.5–108.5%). Pilot research using HaCaT spontaneously immortalized human keratinocytes in a model for wound healing was performed, indicating the usefulness of the method in studies of disturbances in proline metabolism. The developed method addresses the need to determine the intracellular concentration of three key amino acids and can be used routinely in targeted mammalian cell culture metabolomics research.
Agnieszka Klupczynska; Magdalena Misiura; Wojciech Miltyk; Ilona Oscilowska; Jerzy Palka; Zenon J. Kokot; Jan Matysiak. Development of an LC-MS Targeted Metabolomics Methodology to Study Proline Metabolism in Mammalian Cell Cultures. Molecules 2020, 25, 4639 .
AMA StyleAgnieszka Klupczynska, Magdalena Misiura, Wojciech Miltyk, Ilona Oscilowska, Jerzy Palka, Zenon J. Kokot, Jan Matysiak. Development of an LC-MS Targeted Metabolomics Methodology to Study Proline Metabolism in Mammalian Cell Cultures. Molecules. 2020; 25 (20):4639.
Chicago/Turabian StyleAgnieszka Klupczynska; Magdalena Misiura; Wojciech Miltyk; Ilona Oscilowska; Jerzy Palka; Zenon J. Kokot; Jan Matysiak. 2020. "Development of an LC-MS Targeted Metabolomics Methodology to Study Proline Metabolism in Mammalian Cell Cultures." Molecules 25, no. 20: 4639.
Honey bees have a sting which allows them to inject venomous substances into the body of an opponent or attacker. As the sting originates from a modified ovipositor, it only occurs in the female insect, and this is a defining feature of the bee species that belong to a subclade of the Hymenoptera called Aculeata. There is considerable interest in bee venom research, primarily because of an important subset of the human population who will develop a sometimes life threatening allergic response after a bee sting. However, the use of honey bee venom goes much further, with alleged healing properties in ancient therapies and recent research. The present paper aims to standardize selected methods for honey bee venom research. It covers different methods of venom collection, characterization and storage. Much attention was also addressed to the determination of the biological activity of the venom and its use in the context of biomedical research, more specifically venom allergy. Finally, the procedure for the assignment of new venom allergens has been presented. Las abejas melíferas tienen un aguijón que les permite inyectar sustancias venenosas en el cuerpo de un oponente o atacante. El aguijón es un ovipositor modificado que solo se manifiesta en el insecto hembra, siendo este una característica que define a las especies de abejas que pertenecen al subclado de himenópteros llamada Aculeata. Hay un interés considerable en la investigación del veneno de abeja, principalmente debido a que un porcentaje importante de la población humana desarrollará una respuesta alérgica - a veces mortal - a la picadura de abeja. Sin embargo, el uso del veneno de la abeja melífera abarca mucho más, con presuntas propiedades curativas en terapias antiguas e investigaciones recientes. El presente trabajo tiene como objetivo estandarizar métodos seleccionados para la investigación del veneno de las abejas melíferas. Cubre diferentes métodos de recolección, caracterización y almacenamiento de veneno. También se prestó mucha atención a la determinación de la actividad biológica del veneno y su uso en el contexto de la investigación biomédica, más específicamente la alergia al veneno. Finalmente, se ha presentado el procedimiento para la asignación de nuevos alérgenos de veneno.
Dirk C. De Graaf; Márcia Regina Brochetto Braga; Rusleyd Maria Magalhães De Abreu; Simon Blank; Chris H. Bridts; Luc S. De Clerck; Bart Devreese; Didier G. Ebo; Timothy J. Ferris; Margo M. Hagendorens; Débora Laís Justo Jacomini; Iliya Kanchev; Zenon J. Kokot; Jan Matysiak; Christel Mertens; Vito Sabato; Athina L. Van Gasse; Matthias Van Vaerenbergh. Standard methods for Apis mellifera venom research. Journal of Apicultural Research 2020, 1 -31.
AMA StyleDirk C. De Graaf, Márcia Regina Brochetto Braga, Rusleyd Maria Magalhães De Abreu, Simon Blank, Chris H. Bridts, Luc S. De Clerck, Bart Devreese, Didier G. Ebo, Timothy J. Ferris, Margo M. Hagendorens, Débora Laís Justo Jacomini, Iliya Kanchev, Zenon J. Kokot, Jan Matysiak, Christel Mertens, Vito Sabato, Athina L. Van Gasse, Matthias Van Vaerenbergh. Standard methods for Apis mellifera venom research. Journal of Apicultural Research. 2020; ():1-31.
Chicago/Turabian StyleDirk C. De Graaf; Márcia Regina Brochetto Braga; Rusleyd Maria Magalhães De Abreu; Simon Blank; Chris H. Bridts; Luc S. De Clerck; Bart Devreese; Didier G. Ebo; Timothy J. Ferris; Margo M. Hagendorens; Débora Laís Justo Jacomini; Iliya Kanchev; Zenon J. Kokot; Jan Matysiak; Christel Mertens; Vito Sabato; Athina L. Van Gasse; Matthias Van Vaerenbergh. 2020. "Standard methods for Apis mellifera venom research." Journal of Apicultural Research , no. : 1-31.
Asthma often begins in childhood, although making an early diagnosis is difficult. Clinical manifestations, the exclusion of other causes of bronchial obstruction, and responsiveness to anti-inflammatory therapy are the main tool of diagnosis. However, novel, precise, and functional biochemical markers are needed in the differentiation of asthma phenotypes, endotypes, and creating personalized therapy. The aim of the study was to search for metabolomic-based asthma biomarkers among free amino acids (AAs). A wide panel of serum-free AAs in asthmatic children, covering both proteinogenic and non-proteinogenic AAs, were analyzed. The examination included two groups of individuals between 3 and 18 years old: asthmatic children and the control group consisted of children with neither asthma nor allergies. High-performance liquid chromatography combined with tandem mass spectrometry (LC-MS/MS technique) was used for AA measurements. The data were analyzed by applying uni- and multivariate statistical tests. The obtained results indicate the decreased serum concentration of taurine, L-valine, DL-β-aminoisobutyric acid, and increased levels of ƴ-amino-n-butyric acid and L-arginine in asthmatic children when compared to controls. The altered concentration of these AAs can testify to their role in the pathogenesis of childhood asthma. The authors’ results should contribute to the future introduction of new diagnostic markers into clinical practice.
Joanna Matysiak; Agnieszka Klupczynska; Kacper Packi; Anna Mackowiak-Jakubowska; Anna Bręborowicz; Olga Pawlicka; Katarzyna Olejniczak; Zenon J. Kokot; Jan Matysiak. Alterations in Serum-Free Amino Acid Profiles in Childhood Asthma. International Journal of Environmental Research and Public Health 2020, 17, 4758 .
AMA StyleJoanna Matysiak, Agnieszka Klupczynska, Kacper Packi, Anna Mackowiak-Jakubowska, Anna Bręborowicz, Olga Pawlicka, Katarzyna Olejniczak, Zenon J. Kokot, Jan Matysiak. Alterations in Serum-Free Amino Acid Profiles in Childhood Asthma. International Journal of Environmental Research and Public Health. 2020; 17 (13):4758.
Chicago/Turabian StyleJoanna Matysiak; Agnieszka Klupczynska; Kacper Packi; Anna Mackowiak-Jakubowska; Anna Bręborowicz; Olga Pawlicka; Katarzyna Olejniczak; Zenon J. Kokot; Jan Matysiak. 2020. "Alterations in Serum-Free Amino Acid Profiles in Childhood Asthma." International Journal of Environmental Research and Public Health 17, no. 13: 4758.
Background: Oral squamous cell carcinoma remains a significant worldwide public health challenge, associated with high morbidity and mortality. Treatment of this type of cancer lacks effective medication. Moreover, there are very few specific biomarkers that are useful in early diagnosis or treatment optimisation. Proline metabolism may prove to be of importance in the search for new treatment modalities. Methods: To evaluate the significance of proline metabolism in the development of oral cancer, proline concentration was assessed in oral cancer tissue and normal oral mucosa. The results were compared to the clinical stage and histological grade of the tumours. Moreover, the expression of proteins involved in proline metabolism via proline dehydrogenase/oxidase (PRODH/POX, PPARγ, HIF1-α) was determined. In the next stage of the study, conducted on cell lines of tongue cancer treated with celecoxib, the aforementioned factors involved in proline metabolism were evaluated. Cellular viability and cell proliferation, as well as apoptosis, were also assessed. Results: Our research results indicate that a high intracellular proline concentration and expression of factors involved in its metabolism correlate with the clinical stage and histological grade of oral cancer. Moreover, we are the first researchers to demonstrate that celecoxib can affect proline metabolism, causing an increase in pro-apoptotic factors (PRODH/POX, PPARγ), reducing the expression of HIF-1α and activating apoptosis. Conclusions: Proline metabolism, due to its involvement in the process of apoptosis, can be of great importance in anticancer therapy. It appears that celecoxib, which influences the PRODH/POX pathway, may be a promising therapeutic compound in oral cancer treatment.
Natalia Tołoczko-Iwaniuk; Dorota Dziemiańczyk-Pakieła; Katarzyna Celińska-Janowicz; Ilona Zaręba; Agnieszka Klupczyńska; Zenon Kokot; Beata Klaudia Nowaszewska; Joanna Reszeć; Jan Borys; Wojciech Miltyk. Proline-Dependent Induction of Apoptosis in Oral Squamous Cell Carcinoma (OSCC)—The Effect of Celecoxib. Cancers 2020, 12, 136 .
AMA StyleNatalia Tołoczko-Iwaniuk, Dorota Dziemiańczyk-Pakieła, Katarzyna Celińska-Janowicz, Ilona Zaręba, Agnieszka Klupczyńska, Zenon Kokot, Beata Klaudia Nowaszewska, Joanna Reszeć, Jan Borys, Wojciech Miltyk. Proline-Dependent Induction of Apoptosis in Oral Squamous Cell Carcinoma (OSCC)—The Effect of Celecoxib. Cancers. 2020; 12 (1):136.
Chicago/Turabian StyleNatalia Tołoczko-Iwaniuk; Dorota Dziemiańczyk-Pakieła; Katarzyna Celińska-Janowicz; Ilona Zaręba; Agnieszka Klupczyńska; Zenon Kokot; Beata Klaudia Nowaszewska; Joanna Reszeć; Jan Borys; Wojciech Miltyk. 2020. "Proline-Dependent Induction of Apoptosis in Oral Squamous Cell Carcinoma (OSCC)—The Effect of Celecoxib." Cancers 12, no. 1: 136.
Organic acids are important active small molecules present in venoms and toxins, which have not been fully explored yet. The aim of the study was the determination of organic acids in honeybee venom (HBV) samples by using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Two protocols for sample preparation were employed. A solid-phase extraction was used for the determination of malonic acid, fumaric acid, glutaric acid, and kynurenic acid. A dilute-and-shoot method was optimal for: citric acid, malic acid, and succinic acid. Chromatographic separation was performed using a Synergi Hydro-RP column. Detection was performed on a triple-quadrupole mass spectrometer operating in multiple reaction monitoring mode. Among the analytes, glutaric acid and kynurenic acid were present in HBV samples in the lowest concentrations, whereas citric acid was the most abundant acid in each sample, and accounted for an average of 86 mg/g (8.6%) of the venom dry weight. Organic acids were discussed in terms of function. This is the first study in the available literature that provides specific data on the content of organic acids in HBV using a validated quantitative method.
Magdalena Pawlak; Agnieszka Klupczynska; Zenon J Kokot; Jan Matysiak. Extending Metabolomic Studies of Apis mellifera Venom: LC-MS-Based Targeted Analysis of Organic Acids. Toxins 2019, 12, 14 .
AMA StyleMagdalena Pawlak, Agnieszka Klupczynska, Zenon J Kokot, Jan Matysiak. Extending Metabolomic Studies of Apis mellifera Venom: LC-MS-Based Targeted Analysis of Organic Acids. Toxins. 2019; 12 (1):14.
Chicago/Turabian StyleMagdalena Pawlak; Agnieszka Klupczynska; Zenon J Kokot; Jan Matysiak. 2019. "Extending Metabolomic Studies of Apis mellifera Venom: LC-MS-Based Targeted Analysis of Organic Acids." Toxins 12, no. 1: 14.
Background Hymenoptera venom allergy is one of the most frequent causes of anaphylaxis. In its most severe form, the reaction to wasp and honey bee stings may be life-threatening. Therefore, immediate and proper diagnosis of venom allergy and implementation of suitable therapy are extremely important. Broadening the knowledge on the mechanism of the allergic reaction may contribute to the improvement of both diagnostic and treatment methods. Thus, this study aimed to discover changes in protein expression in serum of patients allergic to Hymenoptera (wasp and honeybee) venom and to point out proteins and peptides involved in the allergic inflammation. Methods Serum proteomic patterns typical to allergic patients and healthy volunteers were obtained with MALDI-TOF (matrix-assisted laser desorption/ionization-time of flight) mass spectrometer. The spectra were processed, analyzed and compared using advanced bioinformatics tools. The discriminative peaks were subjected to identification with liquid chromatography coupled with tandem mass spectrometry. Results This methodology allowed for the identification of four features differentiating between allergy and control groups. They were: fibrinogen alpha chain, coagulation factor XIII chain A, complement C4-A, and inter-alpha-trypsin inhibitor heavy chain H4. All of these proteins are involved in allergic inflammatory response. Conclusions Extending the knowledge of the Hymenoptera venom sensitization will contribute to the development of novel, sensitive and specific methods for quick and unambiguous allergy diagnosis. Understanding the basis of the allergy at the proteomic level will support the improvement of preventive and therapeutic measures.
Eliza Matuszewska; Joanna Matysiak; Anna Bręborowicz; Katarzyna Olejniczak; Zdzisława Kycler; Zenon Kokot; Jan Matysiak. Proteomic features characterization of Hymenoptera venom allergy. Allergy, Asthma & Clinical Immunology 2019, 15, 1 -8.
AMA StyleEliza Matuszewska, Joanna Matysiak, Anna Bręborowicz, Katarzyna Olejniczak, Zdzisława Kycler, Zenon Kokot, Jan Matysiak. Proteomic features characterization of Hymenoptera venom allergy. Allergy, Asthma & Clinical Immunology. 2019; 15 (1):1-8.
Chicago/Turabian StyleEliza Matuszewska; Joanna Matysiak; Anna Bręborowicz; Katarzyna Olejniczak; Zdzisława Kycler; Zenon Kokot; Jan Matysiak. 2019. "Proteomic features characterization of Hymenoptera venom allergy." Allergy, Asthma & Clinical Immunology 15, no. 1: 1-8.
Background The pathophysiology of brain injury following aneurysmal subarachnoid haemorrhage (SAH) is associated with numerous mediators. The aim of the study is to analyse protein changes after SAH in cerebrospinal fluid (CSF) using mass spectrometry (MS). Methods CSF samples were obtained from forty-four control subjects, seven good outcome and ten poor outcome SAH patients. CSF samples were collected at specific time intervals after SAH (days 1, 5 and 10). MALDI-TOF (Matrix Assisted Laser Desorption/Ionization Time-of-Flight) and ClinProTools software were utilised for MS, MS/MS (Mass Spectrometry) spectra collection and analysis. Selected masses were identified. The MALDI-TOF profiling experiments allowed for the targeted selection of potential markers in SAH. The study was performed in three steps by comparison of CSF samples: (1) from the control group and SAH patients (both good and poor outcome groups); (2) collected on days 1, 5 and 10 within the groups of poor SAH and good SAH patients, respectively; (3) from poor outcome SAH and good outcome patients at days 1, 5 and 10. Results 15 new proteins whose CSF level is alternated by SAH presence, SAH treatment outcome and time passed since aneurysm rupture were identified. Conclusions We demonstrated new proteins which might play a role in different stages of subarachnoid haemorrhage and could be a new target for further investigation.
Bartosz Sokół; Bartosz Urbaniak; Bartosz Zaremba; Norbert Wąsik; Zenon Kokot; Roman Jankowski. CSF proteomics of patients with hydrocephalus and subarachnoid haemorrhage. Translational Neuroscience 2019, 10, 244 -253.
AMA StyleBartosz Sokół, Bartosz Urbaniak, Bartosz Zaremba, Norbert Wąsik, Zenon Kokot, Roman Jankowski. CSF proteomics of patients with hydrocephalus and subarachnoid haemorrhage. Translational Neuroscience. 2019; 10 (1):244-253.
Chicago/Turabian StyleBartosz Sokół; Bartosz Urbaniak; Bartosz Zaremba; Norbert Wąsik; Zenon Kokot; Roman Jankowski. 2019. "CSF proteomics of patients with hydrocephalus and subarachnoid haemorrhage." Translational Neuroscience 10, no. 1: 244-253.
The ability of early lung cancer diagnosis is an unmet need in clinical practice. Lung cancer metabolomic analyses conducted so far have demonstrated several abnormalities in cancer lipid profile providing a rationale for further study of blood lipidome of the patients. In the present research, we performed a targeted lipidome screening to select molecules that show promise for early lung cancer detection. The study was conducted on serum samples collected from newly diagnosed, stage I non-small cell lung cancer (NSCLC) patients and non-cancer controls. A high-throughput mass spectrometry-based platform with confirmed interlaboratory reproducibility was used. The analyzed profile consisted of acylcarnitines, sphingomyelins, phosphatidylcholines and lysophosphatidylcholines. Among the assayed lipid species, the significant differences between NSCLC and non-cancer subjects were observed in the group of phosphatidylcholines (PC) and lysophosphatidylcholines (lysoPC), especially in the levels of lysoPC a C26:0; lysoPC a C26:1; PC aa C42:4; and PC aa C34:4. The metabolites mentioned above were used to create a multivariate classification model, which reliability was proved by permutation tests as well as external validation. Our study indicated choline-containing phospholipids as potential lung cancer markers. Further investigations of phospholipidome are crucial to better describe the shifts in metabolite composition occurring in lung cancer patients.
Agnieszka Klupczynska; Szymon Plewa; Mariusz Kasprzyk; Wojciech Dyszkiewicz; Zenon Kokot; Jan Matysiak. Serum lipidome screening in patients with stage I non-small cell lung cancer. Clinical and Experimental Medicine 2019, 19, 505 -513.
AMA StyleAgnieszka Klupczynska, Szymon Plewa, Mariusz Kasprzyk, Wojciech Dyszkiewicz, Zenon Kokot, Jan Matysiak. Serum lipidome screening in patients with stage I non-small cell lung cancer. Clinical and Experimental Medicine. 2019; 19 (4):505-513.
Chicago/Turabian StyleAgnieszka Klupczynska; Szymon Plewa; Mariusz Kasprzyk; Wojciech Dyszkiewicz; Zenon Kokot; Jan Matysiak. 2019. "Serum lipidome screening in patients with stage I non-small cell lung cancer." Clinical and Experimental Medicine 19, no. 4: 505-513.
To comprehensively characterize honeybee venom, royal jelly, propolis, and pollen, by applying advanced analytical and bioinformatics methodologies. Honeybee products (HBP) contain many bioactive components with both beneficial and harmful effects on the human organism. Nevertheless, the overall composition of the HBP remains not fully investigated. Thus, this research is focused on complementary proteomic and metabolomic characterization of biologically active compounds derived from HBP, regarding their toxicological and pharmacological properties. The objectives of the study will be achieved by the application of up to date mass spectrometry techniques. Due to increasing interest in using of HBP in medicine, this project will contribute to improving the safety of HBP‑derived dietary supplements and drugs.
Eliza Matuszewska; Paweł Dereziński; Agnieszka Klupczyńska; Agata Światły-Błaszkiewicz; Szymon Plewa; Jan Lubawy; Arkadiusz Urbański; Grzegorz Rosiński; Zenon Kokot; Jan Matysiak. Characterization of the selected honeybee products based on omics techniques. Journal of Medical Science 2019, 88, 129 -132.
AMA StyleEliza Matuszewska, Paweł Dereziński, Agnieszka Klupczyńska, Agata Światły-Błaszkiewicz, Szymon Plewa, Jan Lubawy, Arkadiusz Urbański, Grzegorz Rosiński, Zenon Kokot, Jan Matysiak. Characterization of the selected honeybee products based on omics techniques. Journal of Medical Science. 2019; 88 (2):129-132.
Chicago/Turabian StyleEliza Matuszewska; Paweł Dereziński; Agnieszka Klupczyńska; Agata Światły-Błaszkiewicz; Szymon Plewa; Jan Lubawy; Arkadiusz Urbański; Grzegorz Rosiński; Zenon Kokot; Jan Matysiak. 2019. "Characterization of the selected honeybee products based on omics techniques." Journal of Medical Science 88, no. 2: 129-132.
Introduction. High-mobility group box 1 (HMGB1) is an alarmin with proinflammatory potential determined by redox status of the cysteines at position 23 and 45. It may also play a role as a biomarker in biological fluids. The aim of this study was the identification of different HMGB1 redox forms in cerebrospinal fluid (CSF) obtained from subarachnoid hemorrhage patients. Material and Methods. 6 CSF samples were collected from aneurysmal subarachnoid haemorrhage patients. Commercially available HMGB1 isoforms served as a positive control. Immunoprecipitation and electrophoretic isolation of HMGB1 protein were performed, then both CSF and control were analyzed using mass spectrometry technique. To distinguish between fully reduced (thiol group at C23 and C45) and disulfide (disulfide bond connecting C23 and C45) HMGB1 forms, top-down sequencing of the spectra was performed. Results. Top-down sequencing analysis allowed to distinguish between HMGB1 isoforms only in commercially available standard without preceding immunoprecipitation and electrophoresis. MALDI spectra differ i.e. on the fully reduced HMGB1 spectrum fragmentation occurs before and beyond C22, which is not present on the disulfide HMGB1 spectrum. Analysis of HMGB1 isolated from CSF obtained from subarachnoid hemorrhage patients gave no results. Conclusions. Top-down sequencing enables to distinguish between redox forms of HMGB1. Electrophoresis and tryptic digestion cannot precede mass spectrometry analysis of redox forms of HMGB1 due to the reduction of disulfide bonds during these processes. Preferred method of isolation of HMGB1 for direct analysis using top-down sequencing mustn’t include protein digestion or degradation.
Agata Światły; Norbert Wąsik; Joanna Hajduk; Eliza Matuszewska; Paweł Dereziński; Bartosz Sokół; Roman Jankowski; Zenon Kokot; Jan Matysiak. Mass spectrometry analysis of redox forms of High-Mobility Group Box-1 Protein in cerebrospinal fluid: initial experience. Journal of Medical Science 2019, 88, 171 -176.
AMA StyleAgata Światły, Norbert Wąsik, Joanna Hajduk, Eliza Matuszewska, Paweł Dereziński, Bartosz Sokół, Roman Jankowski, Zenon Kokot, Jan Matysiak. Mass spectrometry analysis of redox forms of High-Mobility Group Box-1 Protein in cerebrospinal fluid: initial experience. Journal of Medical Science. 2019; 88 (3):171-176.
Chicago/Turabian StyleAgata Światły; Norbert Wąsik; Joanna Hajduk; Eliza Matuszewska; Paweł Dereziński; Bartosz Sokół; Roman Jankowski; Zenon Kokot; Jan Matysiak. 2019. "Mass spectrometry analysis of redox forms of High-Mobility Group Box-1 Protein in cerebrospinal fluid: initial experience." Journal of Medical Science 88, no. 3: 171-176.
Despite of almost a hundred years of research on cancer metabolism, the biological background of cancerogenesis and cancer-related reprogramming of metabolism remains not fully understood. In order to comprehensively and effectively diagnose and treat the deadliest diseases, the mechanisms underlying these diseases have to be discovered urgently. Among the gynecological malignancies, ovarian cancer is the most common cause of death. The aim of the study was to search for potential cancer-related differences in concentrations of metabolites and interactions between them in serum of women with ovarian cancer and benign ovarian tumor in comparison with healthy controls using targeted metabolomics. These metabolites might serve as biomarkers in the future. We used wide spectrum targeted metabolomics to evaluate serum concentrations of metabolites related to ovarian cancer and compared them against benign ovarian tumors and healthy controls. The measurements were performed using high performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry technique in highly-selective multiple reaction monitoring mode. In this study we confirmed our previous findings about the role of histidine and citrulline in ovarian cancer as well as we indicated new lipid compounds (lysoPC a C16:1, PC aa C32:2, PC aa C34:4 and PC aa C 36:6) potentially involved in cancer metabolism. We indicated interesting interactions between metabolites for further in-depth research which could potentially serve as clinically useful biomarkers in future. Moreover, the presented work attempts to visualize a possible 3D-network of relationships between the molecules found to be related to ovarian malignancy.
Szymon Plewa; Agnieszka Horała; Paweł Dereziński; Ewa Nowak-Markwitz; Jan Matysiak; Zenon J. Kokot. Wide spectrum targeted metabolomics identifies potential ovarian cancer biomarkers. Life Sciences 2019, 222, 235 -244.
AMA StyleSzymon Plewa, Agnieszka Horała, Paweł Dereziński, Ewa Nowak-Markwitz, Jan Matysiak, Zenon J. Kokot. Wide spectrum targeted metabolomics identifies potential ovarian cancer biomarkers. Life Sciences. 2019; 222 ():235-244.
Chicago/Turabian StyleSzymon Plewa; Agnieszka Horała; Paweł Dereziński; Ewa Nowak-Markwitz; Jan Matysiak; Zenon J. Kokot. 2019. "Wide spectrum targeted metabolomics identifies potential ovarian cancer biomarkers." Life Sciences 222, no. : 235-244.
Gestational trophoblastic disease (GTD) is a group of highly aggressive, rare tumors. Human chorionic gonadotropin is a common biomarker used in the diagnosis and monitoring of GTD. To improve our knowledge of the pathology of GTD, we performed protein-peptide profiling on the urine of patients affected with gestational trophoblastic neoplasm (GTN). We analyzed urine samples from patients diagnosed with GTN (n = 26) and from healthy pregnant and non-pregnant controls (n = 17) using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Ions were examined in a linear mode over a m/z range of 1000–10,000. All GTN urine samples were analyzed before and after treatment and compared with those of the controls. The statistical analyses included multivariate classification algorithms as well as ROC curves. Urine sample analyses revealed there were significant differences in the composition of the ions between the evaluated groups. Comparing the pre-treatment and group with the pregnant controls, we identified two discriminatory proteins: hemoglobin subunit α (m/z = 1951.81) and complement C4A (m/z = 1895.43). Then, comparing urine samples from the post-treatment cases with those from the non-pregnant controls, we identified the peptides uromodulin fragments (m/z = 1682.34 and 1913.54) and complement C4A (m/z = 1895.43).
Paulina Banach; Paweł Dereziński; Eliza Matuszewska; Jan Matysiak; Hubert Bochyński; Zenon J. Kokot; Ewa Nowak-Markwitz. MALDI-TOF-MS Analysis in the Identification of Urine Proteomic Patterns of Gestational Trophoblastic Disease. Metabolites 2019, 9, 30 .
AMA StylePaulina Banach, Paweł Dereziński, Eliza Matuszewska, Jan Matysiak, Hubert Bochyński, Zenon J. Kokot, Ewa Nowak-Markwitz. MALDI-TOF-MS Analysis in the Identification of Urine Proteomic Patterns of Gestational Trophoblastic Disease. Metabolites. 2019; 9 (2):30.
Chicago/Turabian StylePaulina Banach; Paweł Dereziński; Eliza Matuszewska; Jan Matysiak; Hubert Bochyński; Zenon J. Kokot; Ewa Nowak-Markwitz. 2019. "MALDI-TOF-MS Analysis in the Identification of Urine Proteomic Patterns of Gestational Trophoblastic Disease." Metabolites 9, no. 2: 30.
Introduction. Recent instrumentation and software advancement enabled to develop new, high‑throughput targeted metabolomics methods for in‑depth exploration of metabolome in a quantitative manner.Material and Methods. The presented targeted metabolomics approach allows to analyze both of serum and CSF in the same way, with identical sample preparation procedures. The analyses were carried out using high‑performance liquid chromatography system coupled to triple quadrupole tandem mass spectrometer with electrospray ion source (LC‑ESI‑QqQ‑MS/MS). Results. The applied targeted metabolomics approach enabled to determine a wide panel of metabolites from different chemical classes of compounds including: acylcarnitines, amino acids and biogenic amines, glycerophospholipids, sphingolipids and sum of hexoses. Finally, 148 metabolites in serum and 57 in cerebrospinal fluid were determined.Conclusions. Here we presented the results of successful implementation of the method of analysis of low‑molecular weight compounds in human serum and CSF using targeted metabolomics. The evaluation of selected groups of metabolites resulted in obtaining the mean concentrations of panel of metabolites in serum and CSF, which gives a valuable information about the metabolome of these matrices.
Szymon Plewa; Paweł Dereziński; Jolanta Florczak-Wyspiańska; Karolina Popławska-Domaszewicz; Wojciech Kozubski; Bartosz Sokół; Roman Jankowski; Jan Matysiak; Zenon J. Kokot. LC-MS/MS based targeted metabolomics method for analysis of serum and cerebrospinal fluid. Journal of Medical Science 2019, 88, 12 -20.
AMA StyleSzymon Plewa, Paweł Dereziński, Jolanta Florczak-Wyspiańska, Karolina Popławska-Domaszewicz, Wojciech Kozubski, Bartosz Sokół, Roman Jankowski, Jan Matysiak, Zenon J. Kokot. LC-MS/MS based targeted metabolomics method for analysis of serum and cerebrospinal fluid. Journal of Medical Science. 2019; 88 (1):12-20.
Chicago/Turabian StyleSzymon Plewa; Paweł Dereziński; Jolanta Florczak-Wyspiańska; Karolina Popławska-Domaszewicz; Wojciech Kozubski; Bartosz Sokół; Roman Jankowski; Jan Matysiak; Zenon J. Kokot. 2019. "LC-MS/MS based targeted metabolomics method for analysis of serum and cerebrospinal fluid." Journal of Medical Science 88, no. 1: 12-20.
The project entitled “Study of serum metabolic profiles of patients with non-small cell lung cancer with special emphasis on the smoking status of patients” is a study based on metabolomics, which is the latest of the “omics” technologies and involves a comprehensive analysis of small molecule metabolites of a specific biological sample. High-throughput and sensitive analytical techniques used in metabolomic investigations are powerful tools in the field of oncology and aids understanding what is happening in cancer cells and searching for new cancer markers. The aim of the project is to determine whether lung cancer patients have a distinct serum metabolic profile and whether this profile is associated with patients’ smoking status. The application of liquid chromatography-high-resolution mass spectrometry-based methodology along with advanced statistical methods will enable to select potential molecules that can be useful in early lung cancer detection.
Agnieszka Klupczynska; Mariusz Kasprzyk; Wojciech Dyszkiewicz; Marcin Grabicki; Halina Batura-Gabryel; Zenon J. Kokot; Jan Matysiak. Study of serum metabolic profiles of patients with non-small cell lung cancer with special emphasis on the smoking status of patients. Journal of Medical Science 2019, 88, 62 -65.
AMA StyleAgnieszka Klupczynska, Mariusz Kasprzyk, Wojciech Dyszkiewicz, Marcin Grabicki, Halina Batura-Gabryel, Zenon J. Kokot, Jan Matysiak. Study of serum metabolic profiles of patients with non-small cell lung cancer with special emphasis on the smoking status of patients. Journal of Medical Science. 2019; 88 (1):62-65.
Chicago/Turabian StyleAgnieszka Klupczynska; Mariusz Kasprzyk; Wojciech Dyszkiewicz; Marcin Grabicki; Halina Batura-Gabryel; Zenon J. Kokot; Jan Matysiak. 2019. "Study of serum metabolic profiles of patients with non-small cell lung cancer with special emphasis on the smoking status of patients." Journal of Medical Science 88, no. 1: 62-65.
Ovarian cancer (OC) diagnosis remains a clinical challenge due to lack of early symptoms and insufficient accuracy of the available diagnostic methods. The purpose of this study was to determine whether osteopontin could be useful in differential diagnosis of ovarian tumors. Serum samples from 92 patients qualified for surgical treatment due to ovarian mass were divided into 2 groups according to the histopathological result: OC including borderline ovarian tumors (n = 39) and benign ovarian tumors (BOTs) (n = 53). CA125, HE4 and osteopontin concentrations were measured in all patients. Areas under the receiver operating characteristic curves (AUC of ROC) were used to compare the discriminative ability of the univariate and multivariate diagnostic models. The addition of osteopontin to ROMA significantly improved the diagnostic performance of the test in 3 of the 5 analyses: 1) in the OC vs BOT group (from AUC of 0.955 to 0.975), 2) in premenopausal women OC vs BOT (from AUC of 0.828 to 0.892) and 3) in the FIGO I-II stage OC vs BOT (from AUC of 0.865 to 0.895). It did not alter the diagnostic performance of multifactor tests in the group of postmenopausal women nor in OC FIGO III-IV stage group. Osteopontin was also the best single marker to differentiate between early stage OC and BOTs (AUC of 0.863). Osteopontin improves the diagnostic performance of a multimarker OC diagnostic test and could be useful in differential diagnosis of ovarian tumors, especially in pre-menopausal women and for early stage OC.
Agnieszka Horała; Agata Swiatly; Jakub Lorek; Zenon J Kokot; Jan Matysiak; Ewa Nowak-Markwitz. Assessment of diagnostic utility of multivariate diagnostic models in differential diagnosis of ovarian tumors. Ginekologia Polska 2018, 89, 568 -572.
AMA StyleAgnieszka Horała, Agata Swiatly, Jakub Lorek, Zenon J Kokot, Jan Matysiak, Ewa Nowak-Markwitz. Assessment of diagnostic utility of multivariate diagnostic models in differential diagnosis of ovarian tumors. Ginekologia Polska. 2018; 89 (10):568-572.
Chicago/Turabian StyleAgnieszka Horała; Agata Swiatly; Jakub Lorek; Zenon J Kokot; Jan Matysiak; Ewa Nowak-Markwitz. 2018. "Assessment of diagnostic utility of multivariate diagnostic models in differential diagnosis of ovarian tumors." Ginekologia Polska 89, no. 10: 568-572.
In presented study the amino acid analysis was performed in serum derived from rheumatoid arthritis patients (RA) according to undertaken therapy and classification of physical disability. The results were compared with previously published data. The levels of 31 free amino acids were determined in 50 serum samples derived from RA subjects and 51 controls. The RA patients were divided into two groups according to the therapy (methotrexate/leflunomide, infliximab/adalimumab/etanercept/tocilizumab, prednisolone/NSAID) and classification of physical disability of the patients. Levels of amino acids were measured by LC-MS/MS. The obtained results were subjected to multivariate statistical tests. According to the therapy that was being used, threonine differentiated RA patients treated with methotrexate/leflunomide - infliximab/adalimumab/etanercept/tocilizumab (p = 0.00954) and infliximab/adalimumab/etanercept/tocilizumab - prednisolone/NSAID (p = 0.03109), while tryptophan differentiated RA patients treated with methotrexate/leflunomide - infliximab/adalimumab/etanercept/tocilizumab (p = 0.01723). In the functional classification, arginine differentiated RA samples between class III and IV (p = 0.02332), while glycine differentiated them between class I+II and III of the Steinbrocker functional classification (p = 0.03366). An analysis of the metabolome profile requires the use of validated bioanalytical methods that are strictly dedicated for this purpose. The obtained results are not accidental (p value less than 0.05), and all of the selected amino acids play an important role in inflammation and immune response. It is suggested that studied amino acids can be considered as a markers for diagnosis of RA and monitoring pharmacotherapy of the disease.
Bartosz Urbaniak; Szymon Plewa; Agnieszka Klupczynska; Dorota Sikorska; Włodzimierz Samborski; Zenon J. Kokot. Serum free amino acid levels in rheumatoid arthritis according to therapy and physical disability. Cytokine 2018, 113, 332 -339.
AMA StyleBartosz Urbaniak, Szymon Plewa, Agnieszka Klupczynska, Dorota Sikorska, Włodzimierz Samborski, Zenon J. Kokot. Serum free amino acid levels in rheumatoid arthritis according to therapy and physical disability. Cytokine. 2018; 113 ():332-339.
Chicago/Turabian StyleBartosz Urbaniak; Szymon Plewa; Agnieszka Klupczynska; Dorota Sikorska; Włodzimierz Samborski; Zenon J. Kokot. 2018. "Serum free amino acid levels in rheumatoid arthritis according to therapy and physical disability." Cytokine 113, no. : 332-339.
Ovarian cancer has emerged as one of the leading cause of gynecological malignancies. So far, the measurement of CA125 and HE4 concentrations in blood and transvaginal ultrasound examination are essential ovarian cancer diagnostic methods. However, their sensitivity and specificity are still not sufficient to detect disease at the early stage. Moreover, applied treatment may appear to be ineffective due to drug-resistance. Because of a high mortality rate of ovarian cancer, there is a pressing need to develop innovative strategies leading to a full understanding of complicated molecular pathways related to cancerogenesis. Recent studies have shown the great potential of clinical proteomics in the characterization of many diseases, including ovarian cancer. Therefore, in this review, we summarized achievements of proteomics in ovarian cancer management. Since the development of mass spectrometry has caused a breakthrough in systems biology, we decided to focus on studies based on this technique. According to PubMed engine, in the years 2008–2010 the number of studies concerning OC proteomics was increasing, and since 2010 it has reached a plateau. Proteomics as a rapidly evolving branch of science may be essential in novel biomarkers discovery, therapy decisions, progression predication, monitoring of drug response or resistance. Despite the fact that proteomics has many to offer, we also discussed some limitations occur in ovarian cancer studies. Main difficulties concern both complexity and heterogeneity of ovarian cancer and drawbacks of the mass spectrometry strategies. This review summarizes challenges, capabilities, and promises of the mass spectrometry-based proteomics techniques in ovarian cancer management.
Agata Swiatly; Szymon Plewa; Jan Matysiak; Zenon J. Kokot. Mass spectrometry-based proteomics techniques and their application in ovarian cancer research. Journal of Ovarian Research 2018, 11, 1 -13.
AMA StyleAgata Swiatly, Szymon Plewa, Jan Matysiak, Zenon J. Kokot. Mass spectrometry-based proteomics techniques and their application in ovarian cancer research. Journal of Ovarian Research. 2018; 11 (1):1-13.
Chicago/Turabian StyleAgata Swiatly; Szymon Plewa; Jan Matysiak; Zenon J. Kokot. 2018. "Mass spectrometry-based proteomics techniques and their application in ovarian cancer research." Journal of Ovarian Research 11, no. 1: 1-13.
Gestational trophoblastic neoplasms (GTN) exemplify a rare, mostly curable but highly aggressive disease. It is often associated with a rapid formation of distant metastases and most likely with an intense neoangiogenesis processes. The aim of the study was to analyze markers in serum of patients with GTN before chemotherapy compared to healthy pregnant women. In this study sixteen protein angiogenesis markers were evaluated in serum of 21 patients with GTN before chemotherapy and compared with healthy pregnant women. Markers were measured using BioPlex Pro Human Cancer Biomarker Panel 1 immunoassay. t-Tests and receiver operating characteristic curves were used for statistical analysis. Receiver operator curve analysis identified six proteins (sTIE-2, osteopontin, sIL-6α, sVEGFR-2, sEGFR, PECAM-1) which had sufficient sensitivity and specificity (AUC > 0,70) to distinguish GTN patients before the treatment from pregnant controls. The levels of three proteins (sTIE-2, osteopontin and sIL-6α) were altered in GTN patients before the treatment as compared to healthy controls (p = 0,0112; p = 0,0442; p = 0,0488, respectively) and thereby may serve as potential disease markers. Serum concentration of proteins related to angiogenesis changes in the course of GTN and may appear useful in the diagnostic process of this disease.
Paulina Banach; Paweł Dereziński; Jan Matysiak; Agnieszka Horała; Agata Światły-Błaszkiewicz; Piotr Jasiński; Zenon J. Kokot; Ewa Nowak-Markwitz. Serum angiogenesis profile in gestational trophoblastic neoplasm using multiplex immunoassay. Life Sciences 2018, 211, 25 -30.
AMA StylePaulina Banach, Paweł Dereziński, Jan Matysiak, Agnieszka Horała, Agata Światły-Błaszkiewicz, Piotr Jasiński, Zenon J. Kokot, Ewa Nowak-Markwitz. Serum angiogenesis profile in gestational trophoblastic neoplasm using multiplex immunoassay. Life Sciences. 2018; 211 ():25-30.
Chicago/Turabian StylePaulina Banach; Paweł Dereziński; Jan Matysiak; Agnieszka Horała; Agata Światły-Błaszkiewicz; Piotr Jasiński; Zenon J. Kokot; Ewa Nowak-Markwitz. 2018. "Serum angiogenesis profile in gestational trophoblastic neoplasm using multiplex immunoassay." Life Sciences 211, no. : 25-30.
Metabolomic studies constantly require high throughput screenings, and this drives development and optimization of methods that include more analytes in a single run, shorten the analysis time and simplify sample preparation. The aim of the study was to develop a new simple and fast liquid chromatography-tandem mass spectrometry-based methodology for quantitative analysis of a panel of ten organic acids in urine. The metabolites selected for the study include ten molecules potentially associated with cancer development. Chromatographic separation involved a Phenomenex Synergi Hydro-RP column under gradient conditions. Quantitation of the analytes was performed in multiple reaction monitoring mode under negative ionization. Validation parameters were satisfactory and in line with the international guidelines. The methodology enabled us to analyze urine samples collected from prostate cancer (PC) (n = 49) and benign prostate hyperplasia (BPH) (n = 49) patients. The obtained concentrations were normalized with urinary specific gravity (USG) prior to statistical analysis. Five analytes were quantified in all urine samples and we observed the following USG-normalized concentration ranges: citric acid (146.5 -6339.8), 3-hydroxyisobutyric acid (22.5-431.7), 2-ketoglutaric acid (4.4-334.4), lactic acid (10.1-786.3), succinic acid (4.1-500.5). 3-hydroxyisobutyric acid significantly decreased between two groups of prostate cancer patients: ≥7 Gleason patients and <7 Gleason patients. Quick sample preparation limited to “dilute and shoot” makes the developed methodology a great tool for future metabolomic studies, especially for detecting disturbances in energy metabolism (Krebs cycle) and amino acids metabolism. The research also broadens our knowledge on the alteration of selected organic acids in PC and BPH patients.
Agnieszka Klupczynska; Szymon Plewa; Natalia Sytek; Wojciech Sawicki; Paweł Dereziński; Jan Matysiak; Zenon J. Kokot. A study of low-molecular-weight organic acid urinary profiles in prostate cancer by a new liquid chromatography-tandem mass spectrometry method. Journal of Pharmaceutical and Biomedical Analysis 2018, 159, 229 -236.
AMA StyleAgnieszka Klupczynska, Szymon Plewa, Natalia Sytek, Wojciech Sawicki, Paweł Dereziński, Jan Matysiak, Zenon J. Kokot. A study of low-molecular-weight organic acid urinary profiles in prostate cancer by a new liquid chromatography-tandem mass spectrometry method. Journal of Pharmaceutical and Biomedical Analysis. 2018; 159 ():229-236.
Chicago/Turabian StyleAgnieszka Klupczynska; Szymon Plewa; Natalia Sytek; Wojciech Sawicki; Paweł Dereziński; Jan Matysiak; Zenon J. Kokot. 2018. "A study of low-molecular-weight organic acid urinary profiles in prostate cancer by a new liquid chromatography-tandem mass spectrometry method." Journal of Pharmaceutical and Biomedical Analysis 159, no. : 229-236.