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1
Content available remote

Cooperation of Ca^2+ and pH in regulation of the activity of the 2-oxoglutarate dehydrogenase complex and its components from bovine kidney cortex

100%
Pawełczyk T., Angielski S.
Acta Biochimica Polonica
|
1984
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vol. 31
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issue 3
289-305
2
Content available remote

N-myc downstream regulated 1 gene and its place in the cellular machinery

100%
Kitowska A., Pawełczyk T.
|
2010
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vol. 57
|
issue 1
15-21
EN
The exact function of the protein product of N-myc downstream regulated 1 gene (NDRG1) is unclear. Depending on the tissue type the NDRG1 protein is localized in the cytoplasm, nucleus, mitochondrion or membranes. Moreover, the expression of NDRG1 may be altered by several factors such as hypoxia, heavy metals, DNA damage, hormones, oncogene, and tumor-suppressor genes. A number of studies emphasize the role of NDRG1 in cancerogenesis. Presumably NDRG1 participates in angiogenesis, metastases, and mechanisms leading to anti-cancer drug resistance. This review summarizes current knowledge about the NDRG1 gene and the position of NDRG1 protein in the cellular machinery. The role of NDRG1 in cancer pathogenesis and its possible usefulness as a prognostic factor for patients with cancer is also discussed.
3
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The ultraviolet studies on protein-lipid interaction of a protein kinase C-γ phorbol-binding domain

81%
Kowara R., Gryckiewicz E., Matecki A., Pawełczyk T.
Acta Biochimica Polonica
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1999
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vol. 46
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issue 2
405-417
4
Content available remote

Expression level of adenosine kinase in rat tissues. Lack of phosphate effect on the enzyme activity.

81%
Sakowicz M., Grdeń M., Pawełczyk T.
|
2001
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vol. 48
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issue 3
745-754
EN
In this report we describe cloning and expression of rat adenosine kinase (AK) in Esccherichaia coli cells as a fusion protein with 6xHis. The recombinant protein was purified and polyclonal antibodies to AK were generated in rabbits. Immunoblot analysis of extracts obtained from various rat tissues revealed two protein bands reactive with anti-AK IgG. The apparent molecular mass of these bands was 48 and 38 kDa in rat kidney, liver, spleen, brain, and lung. In heart and muscle the proteins that react with AK antibodies have the molecular masses of 48 and 40.5 kDa. In order to assess the relative AK mRNA level in rat tissues we used the multiplex PCR technique with β-actin mRNA as a reference. We found the highest level of AK mRNA in the liver, which decreased in the order kidney >spleen >lung >heart >brain >muscle. Measurement of AK activity in cytosolic fractions of rat tissues showed the highest activity in the liver (0.58 U/g), which decreased in the order kidney >spleen >lung >brain >heart >skeletal muscle. Kinetic studies on recombinant AK as well as on AK in the cytosolic fraction of various rat tissues showed that this enzyme is not affected by phosphate ions. The data presented indicate that in the rat tissues investigated at least two isoforms of adenosine kinase are expressed, and that the expression of the AK gene appears to have some degree of tissue specificity.
5
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Content available

Purinergic signaling in B cells

81%
Przybyła T., Sakowicz-Burkiewicz M., Pawełczyk T.
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2018
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vol. 65
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issue 1
1-7
EN
Adenosine and adenosine triphosphate are involved in purinergic signaling which plays an important role in control of the immune system. Much data have been obtained regarding impact of purinergic signaling on dendritic cells, macrophages, monocytes and T lymphocytes, however less attention has been paid to purinergic regulation of B cells. This review summarizes present knowledge on ATP- and Ado-dependent signaling in B lymphocytes. Human B cells have been shown to express A1-AR, A2A-AR, A2B-AR and A3-AR and each subtype of P2 receptors. Surface of B cells exhibits two antagonistic ectoenzymatic pathways, one relies on constitutive secretion and resynthesis of ATP, while the second one depends on degradation of adenosine nucleotides to nucleosides and their subsequent degradation. Inactivated B cells remain under the suppressive impact of autocrine and paracrine Ado, whereas activated B lymphocytes increase ATP release and production. ATP protects B cells from Ado-induced suppression and exerts pro-inflammatory effect on the target tissues, and it is also involved in the IgM release. On the other hand, Ado synthesis is necessary for optimal development, implantation and maintenance of the plasmocyte population in bone marrow in the course of the primary immune response. Moreover, Ado plays an important role in immunoglobulin class switching, which is a key mechanism of humoral immune response. Disruption of purinergic signaling leads to severe disorders. Impairment of Ado metabolism is one of the factors responsible for common variable immunodeficiency. There are several lines of evidence that dysfunction of the immune system observed during diabetes may in part depend on disrupted ATP and Ado metabolism in the B cells.
6
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The dependence of electrophoretic and spectroscopic properties of the pyruvate dehydrogenase complex on mono- and divalent ions

81%
Pawełczyk T., Easom R. A., Olson M. S.
Acta Biochimica Polonica
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1994
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vol. 41
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issue 1
63-72
7
Content available remote

Expression in Escherichia coli of human ARHGAP6 gene and purification of His-tagged recombinant protein.

81%
Ochocka A.-M., Czyżewska M., Pawełczyk T.
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2003
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vol. 50
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issue 1
239-247
EN
In this report we describe cloning and expression of human Rho GTPase activating protein (ARHGAP6) isoform 4 in Escherichia coli cells as a fusion protein with 6xHis. We cloned the ARHGAP6 cDNA into the bacterial expression vector pPROEX-1. Induction of the 6xHis-ARHGAP6 protein in BL21(DE3) and DH5α cells caused lysis of the cells irrespective of the kind of culture medium used. Successful expression of the fusion protein was obtained in the MC4100Δibp mutant strain lacking the small heat-shock proteins IbpA and IbpB. Reasonable yield was obtained when the cells were cultured in Terrific Broth + 1% glucose medium at 22°C for 16 h. The optimal cell density for expression of soluble 6xHis-ARHGAP6 protein was at A_600 about 0.5. Under these conditions over 90% of the fusion protein was present in a soluble form. The 6xHis-ARHGAP6 protein was purified to near homogeneity by a two step procedure comprising chromatography on Ni-nitrilotriacetate and cation exchange columns. The expression system and purification procedure employed made it possible to obtain 1-2 mg of pure 6xHis-ARHGAP6 protein from 300 ml (1.5 g of cells) of E. coli culture.
8
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Comparison of the kinetic properties of the pyruvate dehydrogenase complex from pig kidney cortex and medulla

80%
Pawełczyk T., Olson M. S.
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vol. 40
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issue 3
411-419
9
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Expression level of Ubc9 protein in rat tissues.

71%
Gołębiowski F., Szulc A., Sakowicz M., Szutowicz A., Pawełczyk T.
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2003
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vol. 50
|
issue 4
1064-1073
EN
Ubc9 is a homologue of the E2 ubiquitin conjugating enzyme and participates in the covalent linking of SUMO-1 molecule to the target protein. In this report we describe a simple and efficient method for obtaining pure human recombinant Ubc9 protein. The purified Ubc9 retained its native structure and was fully active in an in vitro sumoylation assay with the promyelocytic leukaemia (PML) peptide as a substrate. In order to better understand the physiology of Ubc9 protein we examined its levels in several rat tissues. Immunoblot analyses performed on tissue extracts revealed quantitative and qualitative differences in the expression pattern of Ubc9. The Ubc9 protein was present at a high level in spleen and lung. Moderate level of Ubc9 was detected in kidney and liver. Low amount of Ubc9 was observed in brain, whereas the 18 kDa band of Ubc9 was barely visible or absent in heart and skeletal muscle. In heart and muscle extracts the Ubc9 antibodies recognized a 38 kDa protein band. This band was not visible in extracts of other rat tissues. A comparison of the relative levels of Ubc9 mRNA and protein indicated that the overall expression level of Ubc9 was the highest in spleen and lung. In spleen, lung, kidney, brain, liver and heart there was a good correlation between the 18 kDa protein and Ubc9 mRNA levels. In skeletal muscle the Ubc9 mRNA level was unproportionally high comparing to the level of the 18 kDa protein. The presented data indicate that in the rat the expression of the Ubc9 protein appears to have some degree of tissue specificity.
10

Ocena potencjału proliferacji i zdolności formowania kolonii in vitro przez komórki izolowane z miazgi ludzkiego zęba (DPSC) oraz brodawki wierzchołkowej (SCAP) hodowane w warunkach standardowych oraz stymulujących różnicowanie

71%
Chomik E., Bautembach-Koberda P., Schütz K., Pawełczyk T., Maciejewska I.
Dental Forum
|
2013
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vol. 41
|
issue 2
15-24
EN
Introduction. Recent data point to unerupted third molars as a promising source of stem cells (SC). Tooth derived SC are clonogenic and present a capacity for self-renewal and colony formation. Additionally, an environmental stimulation induces an in vitro differentiation of SC into multiple lineages, including odontoblasts. Aim of the study. The aim of the study was to evaluate the in vitro potential for proliferation and colony formation by stem cells derived from both dental pulp and apical papilla cultured in both standard medium (control and primary group) and medium modified with ingredients that stimulate mineralization (experimental group). Material and methods. Right after odontectomy the dental pulp and apical papilla were digested with dispase and collagenase type I. DPSCs and SCAPs were sorted using anti STRO-1, CD146, CD34, CD45 antibodies by means of the MACS method. Thereafter, the cells from the initial and control groups were cultured in a standard medium. The medium of the experimental group was additionally modified with ingredients that stimulated mineralization. To assess the cells commitment, the rate of proliferation and colony formation were examined. Results. The analysis showed that SCAPs from all the examined groups proliferated faster and formed more numerous and larger colonies compared to DPSCs. Environmental stimulation reduced proliferation and the ability to form colonies in both the DPSCs and SCAPs lineages. Conclusion. Faster proliferation and a higher ability to form colonies indicates the lower commitment of SCAPs compared to DPSCs. Additionally, the slower proliferation of stem cells from the experimental group suggests their more advanced commitment and differentiation. Although the SCAPs and DPSCs present different degrees of maturation, both cell lineages seem to be promising sources of stem cells.
PL
Wstęp. Niewyrznięte zęby mądrości są źródłem komórek macierzystych, które przy odpowiedniej stymulacji środowiska hodowlanego in vitro różnicują się w liczne linie rozwojowe m.in. odontoblasty, neurocyty, chondrocyty, adipocyty. Cel pracy. Porównanie in vitro potencjału proliferacji i tworzenia kolonii przez komórki izolowane z miazgi ludzkiego zęba (DPSC) oraz brodawki wierzchołkowej (SCAP), hodowanych w środowisku stymulującym mineralizację (grupa doświadczalna) oraz w warunkach standardowych (grupa wyjściowa i kontrolna). Materiał i metody. Po zabiegu odontektomii miazgę komorową oraz brodawkę wierzchołkową trawiono roztworem dyspazy i kolagenazy typu I, a pozyskane komórki hodowano w pożywce αMEM uzupełnionej surowicą bydlęcą, L-glutaminą oraz antybiotykami. Pożywkę w grupie doświadczalnej dodatkowo wzbogacano składnikami stymulującymi mineralizację. Mezenchymalny fenotyp komórek określono metodą MACS z użyciem przeciwciał przeciwko STRO-1 i CD146, a komórki hematopoetyczne i leukocyty eliminowano selekcją negatywną z użyciem przeciwciał przeciwko CD34 i CD45. Potencjał proliferacyjny szacowano pomiarem szybkości rozkładu soli tetrazolowej do formazanu, natomiast zdolność tworzenia kolonii oceniono barwieniem Giemsy. Wyniki. Analiza hodowli komórek DPSC i SCAP wykazała, że komórki SCAP we wszystkich badanych grupach dzieliły się szybciej oraz tworzyły większe i bardziej liczne kolonie w porównaniu z komórkami DPSC. Ponadto, promineralizacyjna stymulacja środowiska hodowlanego powodowała obniżenie zdolności formowania kolonii i spadek potencjału proliferacji w obu liniach komórkowych. Wnioski. Wysoki potencjał proliferacji oraz zdolność tworzenia licznych kolonii przez komórki SCAP dowodzi ich niższej dojrzałości komórkowej w porównaniu do komórek DPSC. Jednak zarówno SCAP jak i DPSC hodowane w środowisku promineralizacyjnym ulegają szybszemu różnicowaniu komórkowemu o czym świadczy ich niższy potencjał do samoodnawiania i spowolnienie procesów proliferacyjnych. Pomimo różnic w dojrzałości komórkowej komórek pochodzących z obu linii komórkowych, zarówno miazga zęba jak i brodawka wierzchołkowa niewyrzniętych zębów ósmych są cennym źródłem mezenchymalnych komórek macierzystych.
11
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Suppression of ID1 expression in colon cancer cells increases sensitivity to 5-fluorouracil

71%
Przybyła T., Sakowicz-Burkiewicz M., Maciejewska I., Bielarczyk H., Pawełczyk T.
|
2017
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vol. 64
|
issue 2
315-322
EN
Adjuvant chemotherapy with 5-fluorouracil remains the basic treatment for patients with advanced colorectal carcinoma. The major obstacle in successful treatment is the ability of CRC cells to acquire chemoresistance. Here we examined the impact of ID1 silencing on the sensitivity of CRC cells to 5-FU. To suppress ID1 expression in HT-29 and HCT-116 cells the cells were transduced with a lentiviral vector carrying the ID1 silencing sequence. Cells with silenced ID1 showed altered expression of epithelial and mesenchymal markers and exhibited increased proliferation rate compared to the parental cells. HCT-116 cells with suppressed ID1 became sensitized to 5-FU and this was not observed in HT-29 cells. Silencing ID1 resulted in altered expression of genes encoding enzymes metabolizing 5-FU. HT-29 cells with suppressed ID1 had significantly reduced mRNA level for thymidine phosphorylase, uridine-cytydine kinase 2 and dihydropyrimidine dehydrogenase. ID1 suppression in HCT-116 cells resulted in an increase of mRNA level for thymidine phosphorylase, thymidine kinase and uridine-cytydine kinase 2 with concurrent drop of dihydropyrimidine dehydrogenase and thymidylate synthetase mRNA levels. In conclusion, ID1 expression impacts the sensitivity of colon cancer cells to 5-FU and may be considered as a potential predictive marker in CRC treatment.
12
Content available remote

Expression, puritication and kinetic properties of human recombinant phospholipase C δ3

70%
Pawełczyk T., Matecki A.
Acta Biochimica Polonica
|
1997
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vol. 44
|
issue 2
221-229
13
Publication available in full text mode
Content available

Expression profile of circulating microRNAs in patients with ischemic heart failure with moderately reduced left ventricular ejection fraction – pilot study

62%
Kaufmann D., Daniłowicz-Szymanowicz L., Sakowicz-Burkiewicz M., Szwoch M., Pawełczyk T., Raczak G.
European Journal of Translational and Clinical Medicine
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2019
|
vol. 1
|
issue 2
53-57
EN
Introduction: Heart failure (HF) is a growing global pandemic that affects millions of people around the world. Despite the progress in medicine, diagnosis and treatment of HF remains problematic. Recently, noncoding micro ribonucleic acids called miRNAs have become significant in the diagnosis and stratification of HF risk. Aim: The aim of this study was the attempt to identify the profile of circulating miRNAs specific for ischemic HF with moderately reduced left ventricular ejection fraction (HFmrEF). Methods and Results: A number of changes in the miRNA profile can characterise patients with ischemic HFmrEF. This is a pilot study before further research on a larger group of patients. Conclusions: Using the quantitative reverse transcription-polymerase chain reaction (qRT-PCR), serum levels of 84 miRNA were measured and compared between a patient with ischemic HFmrEF and a healthy volunteer. Analysis reveals a down-regulation of let-7f-5p and miR-1-3p, as well as up-regulation of miR-100-5p, miR-10b-5p, miR-125a-5p, miR-140-5p, miR-144-3p, miR-149-5p, miR-15b-5p, miR-183-5p, miR-208b-3p, miR-224-5p, miR-26b-5p, miR-27b-3p, miR-302a-3p, miR-320a, miR-7-5p, miR-99a-5p.
14
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Abnormal FHIT gene transcript and c-myc and c-erbB2 amplification in breast cancer.

62%
Kowara R., Gołębiowski F., Chrzan P., Skokowski J., Karmolinski A., Pawełczyk T.
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2002
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vol. 49
|
issue 2
341-350
EN
Searching for ways to improve the characterization of breast cancer we examined the relationship between the status of the FHIT gene transcript and amplification of c-myc and the c-erbB2 oncogene. Abnormal FHIT transcript was detected in 32 of 79 cancers examined. The presence of Fhit protein estimated by Western blots was evident only in cancers exhibiting a normal-sized FHIT transcript. This indicates that abnormal FHIT transcripts observed in our study did not encode any Fhit protein or the amount of such protein was very low. There was no association between the presence of aberrant FHIT gene transcript with age, tumor size, estrogen and progesterone receptor status, local metastases and histological grading. However, the abnormalities in FHIT gene transcripts were observed with different frequency depending on the histopathological type of the tumor. The aberrant FHIT transcript was detected in 60% of lobular cancers and only in 28% of ductal cancers. Analyzing the occurrence of c-myc and c-erbB2 amplification and the presence of aberrant FHIT gene transcripts we found that the aberrant FHIT transcript more frequently occurred in tissues with c-myc amplification. There was a significant (P <0.05) correlation between the occurrence of the aberrant FHIT gene transcript with accompanying c-myc amplification and positive lymph node status. However, in order to evaluate the predictive value of these findings in breast cancer, an extended clinical follow up will be necessary.
15
Publication available in full text mode
Content available

Expression profile of circulating microRNAs in patients with ischemic heart failure with moderately reduced left ventricular ejection fraction – pilot study

62%
Kaufmann D., Daniłowicz-Szymanowicz L., Sakowicz-Burkiewicz M., Szwoch M., Pawełczyk T., Raczak G.
|
2019
|
vol. 1
|
issue 2
53-57
EN
Introduction Heart failure (HF) is a growing global pandemic that affects millions of people around the world. Despite the progress in medicine, diagnosis and treatment of HF remains problematic. Recently, noncoding micro ribonucleic acids called miRNAs have become significant in the diagnosis and stratification of HF risk. Aim The aim of this study was the attempt to identify the profile of circulating miRNAs specific for ischemic HF with moderately reduced left ventricular ejection fraction (HFmrEF). Methods and Results A number of changes in the miRNA profile can characterise patients with ischemic HFmrEF. This is a pilot study before further research on a larger group of patients. Conclusions Using the quantitative reverse transcription-polymerase chain reaction (qRT-PCR), serum levels of 84 miRNA were measured and compared between a patient with ischemic HFmrEF and a healthy volunteer. Analysis reveals a down-regulation of let-7f-5p and miR-1-3p, as well as up-regulation of miR-100-5p, miR-10b-5p, miR-125a-5p, miR-140-5p, miR-144-3p, miR-149-5p, miR-15b-5p, miR-183-5p, miR-208b-3p, miR-224-5p, miR-26b-5p, miR-27b-3p, miR-302a-3p, miR-320a, miR-7-5p, miR-99a-5p.
16
Content available remote

Isozymes delta of phosphoinositide-specific phospholipase C

60%
Pawełczyk T.
Acta Biochimica Polonica
|
1999
|
vol. 46
|
issue 1
91-98
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