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J Wysocka

Publications and source records attributed to J Wysocka.

At least 19 recordsLinked to original sources
PubMed2006

Flow cytometric analysis of HLA-DR antigen in conjunctival epithelial cells of patients with cystic fibrosis.

PURPOSE: Cystic fibrosis (CF) is an autosomal-recessive genetic disorder. The disease affects all secretory epithelia including the eye and belongs to the group of ocular surface epithelial diseases, termed keratoconjunctivitis sicca that develop in dry eye. In the pathogenesis of dry eye, inflammation plays a crucial role. The aim of this study was to investigate the expression of HLA-DR on conjunctival epithelial cells from patients with CF. MATERIALS AND METHODS: Twenty-five patients with CF and 25 normal subjects underwent ocular examination. Tear film break-up time (TBUT), Schirmer test, lissamine green staining, and conjunctival impression cytology were carried out. Cells were processed for flow cytometry, by using monoclonal antibodies to HLA-DR. RESULTS: The Schirmer test and TBUT scores were significantly lower in CF patients compared with controls. A significant increase of HLA-DR expression on epithelial cells was found in patients with CF compared with normal eyes. The Schirmer and TBUT test were positively correlated with HLA-DR expression for the percentage of cells. CONCLUSION: These results suggest that conjunctival epithelial cells play an important proinflammatory role in ocular changes in CF patients. Our findings confirm the presence of an inflammatory background and the immune nature of this disease. HLA-DR measurement might be a useful method for monitoring of inflammatory processes in the conjunctiva and could be helpful in the use of anti-inflammatory drugs in the treatment of ocular findings in CF patients.

Adolescent↗
PubMed2006

Acute lymphoblastic leukaemia cells express CCR7 but not higher amounts of IL-10 after CD40 ligation.

OBJECTIVE: Production of cytokines that support T-cell activation and proliferation and migration to lymph nodes is one of the most important terms of cancer vaccine development. In previous studies we and others used CD40 ligation to obtain higher expression of co-stimulatory and adhesion molecules on leukaemic cells from children with acute lymphoblastic leukaemia (ALL). This time we assess the cytokine and chemokine gene expression profile in CD40-stimulated ALL cells. MATERIAL AND METHODS: Malignant cells from 25 children with BCP-ALL were stimulated (or not) with huCD40LT and rIL-4 for 96 h. Eleven different molecule, cytokine and chemokine mRNAs levels (CCR7, IL-23, TGF-beta-IP, IFN-gamma, IL-10, CD1a, CD40, CD54, CD80, CD83, CD86) were determined using the real-time PCR technique with TaqMan chemistry using ready-to-use low-density arrays for gene expression by Applied Biosystems. RESULTS: 1) Increases in mRNA levels for CD40, CD54 and CD80 after CD40L and IL-4 stimulation were observed, 2) CCR7 mRNA expression was higher after CD40 ligation than before the culture (p = 0.002), 3) IL-10 mRNA expression was higher after the culture with medium than before the culture (p = 0.01). CONCLUSIONS: The results show that leukaemia-derived dendritic cells obtained with CD40 ligation express CCR7 - chemokine is involved in migration to lymph nodes and does not produce higher amounts of IL-10, a potent immunosuppressive cytokine. Our preclinical findings could be used in the design of immunotherapy trials for the treatment of children with ALL.

CD40 Antigens↗
PubMed2004

Northern Polish population data and forensic usefulness of 15 autosomal STR loci.

Allele frequencies for the 15 STR loci in the AmpF/STR Identifiler kit (Applied Biosystems): D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S539, D2S1338, D19S433, VWA, TPOX, D18S51, D5S818, and FGA were determined in a sample of 145 unrelated individuals living in the northern part of Poland. The values of heterozygosity, polymorphic information content (PIC), power of discrimination (PD), power of exclusion (PE), paternity index (PI) and matching probability (pM)were calculated.

DNA Fingerprinting↗
PubMed2001

Lymphocyte subpopulations in hypertrophied adenoid in children.

Adenoid hypertophy is a common feature of childhood. It is currently accepted that it is caused by the antigen-stimulated increased activity of lymphocyte B (D. Bani, O. Gallo, O. Fini-Storchi, Intraepithelial lymphocyte subpopulations and dendritic accessory cells in normal and hypertrophic adenoids, Laryngoscope 10 (1994) 869-873). The adenoid decreases its size with age but the accompanying function alterations are not fully understood (L. Zawadzka-Glos, M. Chmielik, M. Wasik, Cell mediated response in hypertrophied tonsils in children, Nowa Pediatr. 4 (1997) 12-13). The understanding of the adenoid structure that undergoes some changes during the growth period is essential for evaluation of indications for adnoidectomy and assessment of its potential results. The aim of this study was to evaluate lymphocyte subpopulations in adenoid according to age. The analysed material was adenoids removed on the grounds of hypertrophy, which caused obstructive symptoms and/or otitis media with effusion onset. In the present study, we did not find any statistically significant differences among lymphocytes B, Th, and Ts subpopulations, respectively, in the adenoids of any of the age groups. We have found a statistically significant CD3(+) HLA-DR(+) cell percentage decrease in the group of children from 5 to 10 and above 10 years of age, respectively. We have also found a statistically significant increase in the percentage of NK (CD3(-) CD16(+) 56(+)) lymphocytes in relation to age. On the grounds of the current study, it may be stated that some changes in lymphocyte subpopulations in the adenoid take place with age.

Adenoids↗
PubMed2001

Developmental and cell-cycle regulation of Caenorhabditis elegans HCF phosphorylation.

HCF-1 is a mammalian protein required for cell proliferation. It is also involved in transcriptional activation of herpes-simplex-virus immediate-early gene transcription in association with the viral transactivator VP16. HCF-1 and a related protein called HCF-2 possess a homologue in Caenorhabditis elegans that can associate with and activate VP16. Here, we demonstrate developmental regulation of C. elegans HCF (CeHCF) phosphorylation: a hyperphosphorylated form of CeHCF is present in embryos, whereas a hypophosphorylated form is present in L1 larvae. The phosphorylation patterns of endogenous CeHCF in worms and ectopically synthesized CeHCF in mammalian cells are remarkably similar, suggesting that the way CeHCF can be recognized by kinases is conserved in animals. Phosphorylation-site mapping of endogenous CeHCF, however, revealed that phosphorylation occurs at four clustered sites in the region of the protein that is not highly conserved among HCF proteins and is not required for VP16-induced complex formation. Indeed, phosphorylation of either CeHCF or human HCF-1 appears to be dispensable for association with VP16. All four CeHCF phosphorylation sites match the consensus recognition site for the cell-cycle kinases CDC2 and CDK2. Consistent with this similarity and with the developmental phosphorylation of CeHCF in C. elegans embryos, CeHCF phosphorylation is cell-cycle-regulated in mammalian cells.

Alanine↗
PubMed2001

Loss of HCF-1-chromatin association precedes temperature-induced growth arrest of tsBN67 cells.

Human HCF-1 is a large, highly conserved, and abundant nuclear protein that plays an important but unknown role in cell proliferation. It also plays a role in activation of herpes simplex virus immediate-early gene transcription by the viral regulatory protein VP16. A single proline-to-serine substitution in the HCF-1 VP16 interaction domain causes a temperature-induced arrest of cell proliferation in hamster tsBN67 cells and prevents transcriptional activation by VP16. We show here that HCF-1 is naturally bound to chromatin in uninfected cells through its VP16 interaction domain. HCF-1 is chromatin bound in tsBN67 cells at permissive temperature but dissociates from chromatin before tsBN67 cells stop proliferating at the nonpermissive temperature, suggesting that loss of HCF-1 chromatin association is the primary cause of the temperature-induced tsBN67 cell proliferation arrest. We propose that the role of HCF-1 in cell proliferation is to regulate gene transcription by associating with a multiplicity of DNA-bound transcription factors through its VP16 interaction domain.

Animals↗
PubMed2001

[Granules of neutrophils].

The exocytosis of cytoplasmic granules plays a most important role in the regulation of many neutrophils functions: adhesion, phagocytosis, killing of bacteria and interaction with endothelial cells. Neutrophils contain following types of granules: azurophilic (primary) granules, specific (secondary) granules, gelatinase-rich tertiary granules and secretory vesicles. Neutrophil granules may be classified on the basis of their size, morphology, density or with reference to a given protein.

Cell Adhesion↗
PubMed2001

[Lymphocyte subpopulations in the adenoids analyzed by flow cytometry].

Adenoid hypertrophy is a common problem of childhood. The peak occurrence of adenoid hypertrophy correlates with the onset of serous otitis media. The aim of the study was to evaluate the lymphocyte subpopulations in the adenoids of children with serous otitis media. The adenoids obtained from 65 children at the age of 1 to 17 years were examined. Two groups were studied: 1) children with persistent effusion in the middle ear, 2) children with adenoid hypertrophy without a history of otitis media. The percentage of the lymphocytes B in the adenoids was similar in both groups. However, the percentage of lymphocytes T (p < 0.04), and CD4+ (p < 0.005) was significantly lower in adenoids of children with serous otitis media. The percentage of lymphocytes T HLA-DR+ was slightly higher in this group, but the percentage of lymphocytes T CD25+ was the same in both groups. The quantitative differences in the lymphocyte subpopulations of adenoids may be the predisposing factor in serous otitis media.

Adenoids↗
PubMed2001

Cellular immunity in pediatric uveitis.

To evaluate cellular immunity in children with uveitis, we used the flow cytometry method to study the T lymphocyte subpopulation in peripheral blood. Cytometric analysis of peripheral blood lymphocytes using fluorochrome-marked monoclonal antibodies was carried out in 29 children aged 6-18 years with parasitic uveitis (group A) and with uveitis of another etiology (group B). The control group consisted of 28 healthy children aged 6-15 years. A substantially higher percentage of B lymphocytes (19.0 +/- 6.03) was observed in the children with parasitic uveitis than was observed in the healthy children. Moreover, the absolute values were higher in groups A (1.60 +/- 0.53) and B (1.81 +/- 0.40) than in controls (1.50 +/- 0.38). The lowest percentage of CD4+ lymphocytes (34.25 +/- 6.06) and the highest percentage of CD8+ lymphocytes (31.14 +/- 6.50) was found in children with parasitic uveitis. The lowest index CD4+/CD8+ (1.29 +/- 0.53) in the group of children with parasitic uveitis reflected severe disorders in response regulation. However, the percentage values of natural killer cells was lower in the two groups of patients than in controls. These results suggest that quantitative changes in the subpopulation of peripheral blood lymphocytes are more pronounced in parasitic uveitis.

Adolescent↗
PubMed2001

[Diagnostic usefulness of cystatin C measurement].

Cystatin C is a new endogenous marker of renal function whose serum concentration correlates better with glomerular filtration rate than creatinine. Cystatin C has been suggested to be a marker of renal function). Cystatin C is a cysteine proteinase inhibitor. It is produced at a constant rate in all investigated nucleated cells, its gene is a housekeeping type. Cystatin C is freely filtered in the renal glomeruli and reabsorbed and catabolised in the proximal tubules. Reference intervals have been determined for cystatin C in adults and in children older than one year without gender differences. Several studies including adults and children with different renal diseases with various kidney function have suggested serum cystatin C to be a better marker of GFR than serum creatinine. Fully automated assays using particle-enhanced turbidimetry (PETIA) or particle-enhanced nephelometry (PENIA) are available and the assays are precise, rapid and usable in clinical routine practice.

Cystatin C↗
PubMed2001

[Estimation of platelet counts and their morphological parameters in patients infected by borrelia burgdorferi].

Platelet counts and their morphologic parameters in patients with Lyme borreliosis before and after antibiotic therapy (4 weeks of treatment) were estimated. 30 patients aged 17-60, x = 41 were evaluated: 7 with Erythema migrans, 3 patients with neuroborreliosis in the from Lymphocytic meningitis and 20 ones with Lyme arthritis. Control group consisted of 19 healthy persons aged 34-52, x = 43. Hematologic analyser Coulter MAXM was used for testing PLT, MPV, PCT and PDW. The results indicated that patients with Lyme boreliosis have decreasing platelet count with simultaneously increasing their volume in comparison with healthy control. It may result from the involement of platelets in defense mechanisms of infected host. The decrease of platelet count after the antibiotic treatment in comparison with the control group may be the reflection of influence of antibiotic treatment on thrombopoesis.

Adolescent↗
PubMed2001

[Selected parameters of cellular immunity in uveitis in children].

Evaluation of lymphocyte in children with uveitis basing on the study of T lymphocyte subpopulation in the peripheral blood using the flow cytometry method is presented. Cytometric analysis of peripheral blood lymphocytes using fluorochrome-marked monoclonal antibodies was carried out in 29 children, aged 6-18, with parasitic uveitis (group A) and with uveitis of another aetiology (group B). Control group consisted of 28 healthy children aged 6-15. The absolute values of T lymphocytes were higher in groups A (x = 1.60 +/- 0.53) and B (x = 1.81 +/- 0.40) compared to controls (x = 1.50 +/- 0.38). The lowest percentage of CD4+ lymphocytes (x = 34.25 +/- 6.06) and the highest percentage of CD8+ lymphocytes (x = 31.14 +/- 6.50) were revealed in children with parasitic uveitis. The lowest index CD4+/CD8+ (x = 1.29 +/- 0.53) in the group of children with parasitic uveitis reflects severe disorders in response regulation. However, the percentage values of NK cells were lower in the two groups of patients compared to controls. The results allow for the assumption that quantitative changes in the subpopulation of peripheral blood lymphocytes are more pronounced in parasitic uveitis.

Adolescent↗
PubMed2000

Peripheral blood reticulocytes and their reference range values for percentage, absolute count, and immature fraction in children, measured with flow cytometry.

Reticulocyte count by manual method has been the assay traditionally used to evaluate the status of erythropoiesis in hematological disorders with disturbances in erythropoietic activity. However, due to its variability, it is rather a semiquantitative method. Automated reticulocyte counting based on flow cytometry has provided more objective and exact measure of reticulocytes. Besides traditional parameters, such as percentage and absolute number of reticulocytes, automatic reticulocyte counters can detect differences in the amounts of cellular RNA present in immature erythrocytes that reflect their maturational stages and evaluate indices of reticulocyte maturation such as RMI, HFR, IRF. These parameters can be used as the earliest signs of marrow engraftment after autologous or allogeneic bone marrow transplantation. Currently there is no strict agreement between various automated methods of reticulocyte evaluation. Additionally, lack of standardization and quality control materials for this assay compels determination of own, interlaboratory ranges of reference values for new proposed parameters. A group of 102 children aged from 3 months to 18 years with normal hematological parameters was examined. Samples of blood stained supravitally with thiazole orange were analyzed in a flow cytometer. Results for percentage, absolute number and immature reticulocyte fraction expressed as a mean +/- 2SD were: 2.00 +/- 1.56%, 88.8 +/- 68.94 x 10(3)/microliter, and 0.22 +/- 0.16, respectively. A poor correlation was found between IRF and other parameters, suggesting its independent role as a marker of erythropoietic activity. Automated reticulocyte counting will probably improve the diagnosis and monitoring of many hematological diseases.

Adolescent↗
PubMed2000

[Thyroid function in children with diagnosed chronic hepatitis B treated with interferon alpha].

The aim of the study is to estimate thyroid function during interferon alpha therapy in children with chronic hepatitis B. 31 children aged 4-16 years were included in the study. Thyroid function was monitored by estimation of thyroid hormones (FT3, FT4, TSH) and antithyroid antibodies (TPO, ATG) concentration before and after therapy completion. In all children normal thyroid hormones and ATG concentration were found before and after therapy completion. Presence of TPO before IFN treatment in blood serum of 2 girls (6.45%) and in 4 children (12.9%) after IFN therapy were found. Clinical features of thyroid dysfunction were not observed in this group of children during IFN therapy.

Adolescent↗
PubMed2000

[Alpha-fetoprotein: diagnostic value in hepatic disorders].

The serum AFP concentration in man falls rapidly after birth and its synthesis in adult life is normally repressed. However, AFP is synthesized in large amounts by human hepatocellular carcinoma in greater than 70% of patients. Elevation of serum AFP in benign hepatic diseases such as acute and chronic viral hepatitis as well as toxic liver injury is associated with small transient increases in serum AFP. Therefore, quantification of serum alpha-fetoprotein (AFP) has been widely used as a diagnostic marker for hepatocellular carcinoma. Measurement of serum AFP levels have also been used in screening populations at high risk of human hepatocellular carcinoma such as those with cirrhosis or carriers of hepatitis B virus. However the specificity of the screening test in patients with only modestly raised AFP (below 400 ng/ml) is low, and false-positive results are frequent. A wide range of overlap in the distribution of serum AFP levels between hepatocellular carcinoma and chronic liver disease patients were observed mainly among HBsAg (+) patients. Therefore the specificity and predictive value of AFP are lower in HBsAg(+) than in HBsAg(-) patients, especially when AFP is between 25 and 200 ng/ml. In patients with chronic hepatitis B, the analysis of lectin reactivity of AFP has the advantages over quantification of serum AFP to detect HCC-specific variants in serum samples with only moderate raised AFP levels. Measurement of AFP serves as an important tool in the care and management of patients with benign and malignant hepatic disorders.

Adult↗
PubMed2000

[Automatic analysis of reticulocytes--new diagnostic possibilities in hematology].

This review describes a new method of reticulocyte analysis based on flow cytometry, better than manual reticulocyte counting. Reticulocyte percentage, absolute number, indices of maturity (RMI--reticulocyte maturity index, IRF--immature reticulocyte fraction) and others qualitative parameters express the erythropoietic activity of kidneys and bone marrow. The new proposed parameters can improve the diagnosis of anemias and others haematological disorders like leukemia, myelodysplastic syndrome. They also provide exactly and early information about the status of bone marrow after ablative chemotherapy and transplantation. Automated reticulocyte counting can create new potentials for haematology diagnosis.

Flow Cytometry↗
PubMed2000

[New reticulocyte indices and their utility in hematologic diagnosis].

Reticulocyte count in manual method has been the assay traditionally used to evaluate the status of erythropoiesis in haematological disorders with disturbances in erythropoietic activity. But it is a qualitative rather than a quantitative test due to the high variability in manual microscopic method. Automated reticulocyte counting based on flow cytometry has provided much objective and exactly measure of percentage and absolute number of reticulocytes than microscopic method. Besides these two traditional parameters automatic reticulocyte counters can detect differences in the amounts of cellular RNA present in red blood cells which reflects their maturational stages. There are three related parameters which describes the reticulocyte maturation: RMI (reticulocyte maturity index), HFR (high fluorescence reticulocytes), IRF (immature reticulocyte fraction). These parameters can be used as the earliest signs of marrow engraftment after autologous or allogeneic bone marrow transplantations. Reticulocyte cellular indices such as MCVr (mean reticulocyte volume), CHr (reticulocyte hemoglobin content), CHCMr (reticulocyte hemoglobin concentration) entered recently in few automated reticulocyte counters allows for assessment of the functional state of the erythropoiesis in the diagnosis and monitoring of the iron deficiency and rhEpo therapy. Although there is no agreement between various methods of staining and counting of the reticulocytes and lack of standardization materials for this assay automated reticulocyte counting and new reticulocyte parameters probably will improve the diagnosis and monitoring of many haematological diseases.

Erythropoiesis↗