Biomedical subjects

T Wilczok

Publications and source records attributed to T Wilczok.

At least 37 records · Page 2Linked to original sources

PubMed2001

Quantification of P21 mRNA in biopsy specimens of colon tissue at different stages of neoplastic transformation.

Induction of the P21 protein is an integral part of cell growth arrest associated with the cellular response to senescence or damage. Changes in the P21 protein level are often observed in tumour cells and it is suggested, that they correlate with malignancy. We studied the P21 gene expression at the mRNA level in biopsy specimens from colitis ulcerosa, adenomas and adenocarcinomas using TaqMan assay. Our results showed, that quantity of the P21 transcript was evidently lower in the cases with adenocarcinomas than in the cases with colitis ulcerosa or adenomas.

Adenocarcinoma↗

PubMed2001

BCL2 and BAX mRNA concentration profile in fibrillary astrocytoma.

A high level of the BCL2 protein and the lack of apoptosis promoting protein BAX are beginning to be treated as markers of cellular resistance to anti-neoplastic drugs. The object of the study were specimens from stereotactic biopsy of Astrocytoma fibrillare in the central brain area, inaccessible to conventional surgery. The cytological preparations have been evaluated with histopathological and immunohistochemical methods in order to determine the origin of the tumour and assess cell proliferation activity. The molecular analysis conducted in order to determine the sensitivity of the tumour to radio- or chemotherapy included the determination of the number of mRNA BCL2 alpha and beta molecules and of BAX in 1 microg total RNA obtained from microscope slides. A higher expression of BAX than of BCL2-alpha is a prognosis for a positive result of chemo- or radiotherapy. A trace number of mRNA BCL2-beta molecules and a smaller number of mRNA BCL2-alpha molecules than mRNA BAX is a good prognosis for therapy.

Astrocytoma↗

PubMed2001

Concentration profile of mRNA of kinine B1 and B2 receptors in the evaluation of colon pathologies.

Colon pathologies involve the activation of a number of inflammation mediators: cytokines, prostanoids and kinines. The expression patterns of the genes associated with their activation may provide a very sensitive marker of the physiological condition of the cells. This paper demonstrates certain statistically significant differences (p = 0.008) between the expression patterns of kinine B1 and B2 receptor encoding genes for colitis ulcerosa and a control group. The ratio of kinine B1/B2 concentrations changes significantly, on the average from 1.3 for a tissue assessed as healthy to 6.6 for colitis ulcerosa.

Colitis, Ulcerative↗

PubMed2001

Expression of telomerase genes as potential marker of neoplastic changes.

We conducted a quantitative analysis of TERT, TP1 and hTR mRNA expression in various types of endometrial hyperplasia in perimenopausal women, taking advantage of the real-time PCR assay. All women underwent hysterectomy for gynecological reasons. Endometrial dating was determined from the patomorphology of the endometrium and classified into endometrial hyperplasia: simplex, complex and atypica. Our data suggest that only hTR was observed in each normal and hyperplastic endometrium specimens, suggesting that this factor constitutively expressed in endometrium. The results obtained indicate that the expression activity of the TERT subunit changes but not significantly, depending on the stage of development of the hyperplasia.

Adult↗

PubMed2001

Expression of the histone H3 gene in the evaluation of cellular proliferation in colitis ulcerosa.

A comparison of the number of mRNA molecules of histone H3 in 1 microg total RNA extracted from colon sections sampled during colonoscopy was used to evaluate cellular proliferation activity in the rectum and sigmoid, in normal tissue and in colitis ulcerosa. Samples with similar intensity of the disease were selected for the study. Statistically significant differences between both groups of rectal sections were found in the expression of histone H3 encoding genes. The statistically significant result (p = 0.0485) indicates a more active division of cells in the healthy rectum, with no statistically significant differences in the sigmoid (p=0.9575).

Biomarkers↗

PubMed2001

[Histone H3 gene expression level as markers of hyperplasia simplex of endometrium].

SUBJECT: The excessive proliferative activity of endometrial cells drives to hyperplasia, via states: simplex hyperplasia, complex and atypical hyperplasia, and finally preneoplastic state may arise. New molecular markers and their ability to differentiate physiological and pathological states are described. DESIGN: The aim of this study was to evaluate the usefulness of histone H3 gene expression level as a marker of hyperplasia of endometrium. MATERIAL AND METHODS: The level of H3 mRNA was estimated using RT-QPCR method in proliferative phase (n = 5) and endometrial hyperplasia simplex (n = 8). Differences in expression level of H3, were evaluated by Student's t-test (t-test for independent samples). RESULTS: In our experiments the average mRNA H3 expression was significantly higher(p < 0.05, p = 0.03) in case of hyperplasia simplex in comparison to H3 expression in proliferative state of endometrium. CONCLUSION: The results confirm advantage of estimation of H3 mRNA level as a marker of cell proliferation in endometrium.

Adult↗

PubMed2001

[Quantitative (RT-QPCR) analysis of telomerase subunit in normal endometrial tissues].

Telomerase is a specialized ribonucleoprotein polymerase that elongates telomeres. To clarify the molecular mechanisms through which telomerase is activated in normal endometrium, we examined the expression profiles of each telomerase subunit in this tissues. A total of 22 normal human endometrium in various menstrual phases were examined for the expression of each telomerase subunit (hTERT, hTR, TP-1) using RT-QPCR. The hTERT as well as TP-1 subunit had higher expression levels in proliferative than secretory phase endometrium. Expression levels of hTR did not change during menstrual cycle. Our data showed the strong correlation between the level of expression of telomerase in human endometrium and phase of menstrual cycle.

Adult↗

PubMed2001

Short-term changes of serum IL-2 and IL-6 induced by interferon alpha-2b in patients with chronic hepatitis C.

BACKGROUND: The standard therapy of chronic hepatitis C with interferon alpha (IFN alpha) and ribavirin has established but limited efficacy. The prognostic factors of treatment are still under investigation. IL-2 and IL-6 are key cytokines involved in activation of B and T lymphocytes and thus in humoral and cellular responses; they are also deeply involved in generation and maintenance of inflammatory processes. The aim of the study was to evaluate the short-term influence of INF alpha-2b on serum IL-2 and IL-6 levels in sustained responders (SR) and non-responders (NR). MATERIAL AND METHODS: Altogether 12 patients (7 males and 5 females) chronically infected with HCV (anti-HCV positive, HCV-RNA positive by PCR) were enrolled to the study. Patients were treated with IFN 3 MU tiw for 6 months and then they were followed for another 6 months. Five patients responded to the treatment (sustained responders-SR)-Group I, seven patients did not respond (non-responders-NR)-Group II. Serum concentrations of IL-2 and IL-6 were assessed by ELISA before ['0'] and at 1st ['1'], 2nd ['2'], 3rd ['3'], 6th ['4'] and 12th ['5'] hour after the first IFN injection. CONCLUSIONS: Interferon alpha-2b induced short-term increase of serum IL-2 concentrations in SR but not in NR. Serum IL-6 level increased both in SR and NR but this effect was more pronounced and persisted longer in sustained responders.

Adult↗

PubMed2001

Interferon versus interferon and UDCA combined therapy in chronic hepatitis C.

BACKGROUND: Interferon alpha (IFN) has been shown to have established efficacy in the treatment of chronic hepatitis C but its effectiveness is unsatisfactory. Combined therapies with IFN and other antiviral or immunomodulatory drugs are under evaluation. A combination of interferon alpha and ursodeoxycholic acid UDCA has been reported to give better results than interferon alone. The aim of the study was to assess the efficacy of IFN monotherapy versus IFN and UDCA therapy in patients with chronic hepatitis C. MATERIAL AND METHODS: We studied 38 patients (25 males and 13 females) chronically infected with HCV (anti-HCV positive, HCV-RNA positive by PCR). Seventeen of them were treated with IFN 3 MU tiw for 6 months--Group I. The remaining 21 patients were treated with IFN, at the same dosage, plus UDCA (10 mg/kg/day) also for 6 months--Group II. Patients were followed for 6 months. 6 months after the end of therapy, laboratory biochemical parameters, HCV viremia and proportion as well as time to relapse were assessed. CONCLUSIONS: In contrast to previous reports we did not find any differences neither in proportion of HCV reactivation nor in the time of its appearance among patients treated because of chronic hepatitis B with IFN alone or with IFN plus UDCA combined therapy. We also did not find any difference in initial and late response to the treatments in both groups.

DNA, Complementary↗

PubMed2001

Assessment of HBV DNA levels in human plasma and peripheral blood mononuclear cells by TaqMan Q-PCR assay.

Hepatitis B virus DNA (HBV DNA) levels were quantitatively determined by a TaqMan quantitative polymerase chain reaction (TaqMan Q-PCR) in the plasma and in the peripheral blood mononuclear cells (PBMCs) of 120 patients with biopsy proven chronic hepatitis. The number of HBV DNA copies ranged from 10(1) to 10(10)/ml of the blood. There were no significant differences between plasma and PBMCs levels of HBV DNA in neither of HBV infected alone nor HCV co-infected patients. The number of HBV DNA copies was higher in the both compartments in blood of patients infected with HBV only, in comparison to those co-infected with HBV and HCV. In patients chronic hepatitis caused by both viruses (co-infection), decrease of HBV DNA copies in the PBMCs preceded its disappearance from the plasma. Our results suggest that assessment of HBV DNA in PBMCs does not produce important advantage in diagnosis of chronic hepatitis, and it seems to have the prognostic significance of positive outcome among the patients with chronic hepatitis induced by HBV and HCV co-infection. Our results also indirectly indicate that HCV may reciprocally inhibit replication of HBV.

DNA↗

PubMed2001

Increased AST and GGT activity as marker of RT-PCR inhibition in RNA extracts from peripheral blood.

The presence of HCV RNA in PMBC, with simultaneous absence of the virus in the plasma (7.8%), suggests that blood is better material for HCV RNA detection than plasma or serum in the diagnostic procedures of patients with chronic hepatitis C as well as in monitoring the antiviral therapy. We studied 111 patients with chronic hepatitis C (anti-HCV+) and elevated level of at least one biochemical marker: AST, ALAT, GGT, AP and bilirubin. Inhibition of amplification was 2% in plasma and 34% in whole blood samples. We applied modification of extraction to reduce the inhibitory effect on PCR, by introducing additional purification of the RNA extract in the Chomczyński method. After our modification of extraction was applied, inhibition was reduced to 1%. In attempt to identify such inhibitory markers that would label the samples, in which additional RNA extract purification should be applied, we analysed the activity of AST and ALAT enzymes, the key markers for parenchymal liver damage; GGT and AP, the markers for cholestatic hepatitis as well as bilirubin. We observed that the increased GGT and AST activities were correlated with the inhibition of RT-PCR. This correlation was statistically significant; for AST (Mann-Whitney test p = 0.09654 and Kolmogorow-Smirnow test p = 0.01543) and for GGT (Mann-Whitney test p = 0.02419 and Kolmogorow-Smirnow test p = 0.01921).

Alanine Transaminase↗

PubMed2001

Dynamics of HCV replication in patients with chronic hepatitis C during interferon and ribavirin combined therapy.

Objective of the present study was to determine the effect of combined therapy with interferon alfa-2b (3 MU; thrice weekly s.c.) and ribavirin (1200 mg daily p.o.) on the number of copies of positive and negative strands of the hepatitis C virus RNA (HCV RNA) in patients with biopsy-proven chronic hepatitis C. Number of copies of both strands was determined by a TaqMan reverse transcription quantitative polymerase chain reaction (TaqMan RT Q-PCR) in the whole blood before treatment, and 4, 12 or 24 weeks after introduction of the treatment. Before the treatment positive strand of HCV RNA was more frequently detectable and its level was higher compared to that of the negative strand. In several patients, we observed therapy-induced transient appearance or increase in the number of copies of the negative HCV RNA strand. As a result of 24-week treatment, the negative strand of HCV RNA was eliminated from the blood more effectively than the positive strand. Our results suggest that the assessment of dynamics of changes of the positive and negative strands levels of HCV RNA in interferon-naive patients with chronic hepatitis C may be useful in monitoring short-term effectiveness of combined antiviral therapy. They also indicate that TaqMan RT Q-PCR appears to be a valuable tool in monitoring the therapy in these patients.

Antiviral Agents↗

PubMed2001

[Prenatal diagnostics of Toxoplasma gondii invasion by quantitative method of PCR-TaqMann].

Routine serological diagnosis of toxoplasmosis provides high sensitivity, but not high specificity. The high sensivity combined with high specifity offered by PCR-TaqMann as well as the degree of infection led us to investigate the presence and levels of T. gondii DNA in amniotic fluid, maternal and neonatal blood in cases of pregnancy where infection with this agent was suspected. Samples of amniotic fluid and blood were taken from pregnant women. Postnatal blood samples were also taken from their infants. Presence and levels of toxoplasma DNA was investigated using PCR-TaqMann. PCR products were detected by electrophoresis on polyacrylamide gel. The PCR-TaqMann test is highly sensitive, specific and useful method allowing detection of the parasite genome and assessement of its level.

Adult↗

PubMed2001

[Detection of Toxoplasma gondii DNA by PCR in mother's blood, amniotic fluid and child's blood in selected cases of pathological pregnancy].

Prenatal screening of Toxoplasma gondii infection is controversial. The diagnosis is based on serological tests detecting IgM and IgG antibodies against T. gondii, but interpretation of these tests results is often confusing. It is commonly made retrospectively when serological screening indicates a possibility of recent infection. Most women have antibodies against T. gondii and serial testing is required only in monitoring of pregnancies where initial screening is negative. The introduction of Polymerase Chain Reaction (PCR) assay for detection of Toxoplasma gondii DNA in selected cases of pathologic pregnancies has permitted a more accurate and faster diagnosis of congenital toxoplasmosis infections.

Adult↗

PubMed2000

Dopamine-melanin protects against tyrosine nitration, tryptophan oxidation and Ca(2+)-ATPase inactivation induced by peroxynitrite.

The effects of dopamine-melanin (DA-melanin), a synthetic model of neuromelanin, on peroxynitrite-mediated 3-nitrotyrosine formation, oxidation of tryptophan in bovine serum albumin and inactivation of erythrocyte membrane Ca(2+)-ATPase activity were investigated in the absence and in the presence of bicarbonate. DA-melanin inhibited nitration of free tyrosine, loss of tryptophan residues and Ca(2+)-ATPase inactivation by peroxynitrite in a dose dependent manner. In the presence of bicarbonate, this inhibitory effect was lower for nitration and insignificant for oxidative protein modifications. These results suggest that neuromelanin can protect against nitrating and oxidizing action of peroxynitrite but is a worse protector against the peroxynitrite-CO(2) adduct. As peroxynitrite may be a mediator of neurotoxic processes, the obtained results suggest that neuromelanin may be important as a physiological protector against peroxynitrite.

Animals↗

PubMed2000

[Genetic changes and clinical management in familial hypertrophic cardiomyopathy].

Hypertrophic cardiomyopathy (HCM) is phenotypically and genotypically heterogeneous disease of heart. Nine chromosomal loci responsible for this condition have been identified: beta-myosin heavy chain, essential and regulatory myosin light chains, troponin T and I subunits, alpha-tropomosin, cardiac myosin binding protein C, cardiac actin and titin. These genes code for proteins involved in the contraction mechanism or in the control of contraction, therefore HCM has been classified as a disease of cardiac sarcomere. Over 107 mutations have been identified. More then half of them have been detected in the beta-myosin heavy chain gene (beta-MHC). Some mutations in beta-MHC gene are associated with a benign prognosis, other are associated with high incidence of sudden cardiac death (SCD) and severe hypertrophy. Mutations in myosin binding protein C are associated with mild, delayed expression of cardiac hypertrophy and benign prognosis. Mutations in cardiac troponinT are associated with a mild degree of hypertrophy but a high incidence of SCD. Study of genes responsible for HCM will assume role in the context of clinical management of HCM, in particular regarding diagnosis and prognosis patients and families with HCM.

Cardiomyopathy, Hypertrophic↗

PubMed2000

[Expression of VEGF, KDR, p53, E6 HPV16 and HPV18 in vulvar and cervix cancer].

The aim of the study was a comparison of expression of angiogenesis genes: vascular endothelial growth factor (VEGF), KDR, suppressor gene p53, E6-HPV16 and HPV18, in tissue samples of normal, dystrophic, lymph nodes and malignant cancers of vulva and uterine cervix. The results demonstrate that molecular diagnostics of cancers using gene expression profiling indicates the definitive difference in expression profiles of aforementioned genes in tissues of the same malignancy.

Carcinoma, Squamous Cell↗

PubMed2000

[Profiles of expression of genes coding kininogen and kinin receptors as a marker of tissue pathology in cervical cancer coexisting with HPV infection].

Kinins are peptides involved in inflammatory processes, vascular permeability, proliferation and mitogenesis of tumor cells. The majority of kinins actions are mediated through an interaction with cell surface bradykinin receptors BR1 and BR2. Kinins precursor is kininogen (kng). The changes in proteins are initiated by changes in the expression of genes coding these proteins, thus can be a valuable diagnostic markers of malignant processes including cervical carcinoma. The paper presents an analysis of kininogen-kinins receptor genes expression in women treated surgically because of a carcinoma of uterine cervix. Among the studied women in 5 cases previously brachyHDR therapy was applied. In all studied cases HPV 18 infection and in 2 cases a co-infection HPV 16/18 by use of Consensus Primers MY09, MY 11 and type specific primers for HPV 16, 18 was ascertained. In RNA extracts the number of the mRNA copies for kiningen, BR1 and BR2 was assessed using QRT-PCR Taq Man. The higher expression of BR1 than BR2 was marked in the tissue with cancer cells. In the patients after brachytherapy higher expression of BR2 than BR1 mRNA was found. The higher BR1 expression was also shown in iliac lymph nodes in patients with active neoplastic process, opposite to the patients after brachytherapy in whom higher BR2 expression was ascertained. The lack of expression of kng mRNA was found only in 3 specimens. The high expression of kinin receptors especially BR1 in infiltrating carcinoma margin can be a marker of pathology intensity: proliferative potential of neoplasms cells or chronic inflammatory state in the presence of invasive carcinoma.

Adult↗