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Biomedical subjects

F Nowak

Publications and source records attributed to F Nowak.

At least 19 recordsLinked to original sources

IPM-FISH, a new M-FISH approach using IRS-PCR painting probes: application to the analysis of seven human prostate cell lines.

We have developed an alternative multicolor karyotyping technique based on multiplex fluorescence in situ hybridization (M-FISH) and our own optical device with a specific filter set. The most innovative part of our development is the use of interspersed polymerase chain reaction (IRS-PCR) painting probes that show an R-band pattern simultaneous to the combinatorial labeling. This allows us not only to recognize the origin of chromosomal fragments, but to identify the breakpoints as well. We have used this technique to analyze seven cell lines: four prostate cancer cell lines (CA-HPV-10, LNCaP, DU145, and PC3), and three normal transformed epithelial prostate cell lines (PNT1B, PNT2, and PZ-HPV-7). In order to validate our IRS-PCR multiplex FISH (IPM-FISH) technique and to complement the results, we applied comparative genomic hybridization (CGH) and FISH analysis, showing good correlation with the IPM-FISH results. To date, molecular and cytogenetic studies have identified several chromosomal regions that are altered in human prostate cancer; several candidate genes have been suggested. However, reliable markers for predicting the aggressiveness of early prostate cancer are not yet available. Our results show several common, unbalanced rearrangements in the cell lines. These rearrangements are similar to regions already implicated in prostate cancer, validating these cell lines as a good model system.

Cell Line, Transformed↗

Expression of the activated p185erbB2 tyrosine kinase in human epithelial cells leads to MAP kinase activation but does not confer oncogenicity.

Amplification of the c-erbB2 gene and overexpression of p185erbB2 is found in approximately one-third of primary breast and ovarian cancers and also in some colon carcinomas. Moreover, a single point mutation in erbB2(V 664 E) confers transforming potential to erbB2 in NIH3T3 cells, even when expressed at low levels. To examine the transformation potential of erbB2 or erbB2(V-E) in colon epithelial cells, we have transfected a nontumorigenic clone of SW 613-S cells with either wild-type p185erbB2 or mutated p185erbB2(V-E). In contrast to p185erbB2, p185erbB2(V-E) associated constitutively with members of the Shc protein family, leading to phosphorylation of Shc and to stimulation of mitogen-activated protein kinase (MAP kinase). However, constitutive activation of MAP kinase activation in p185erbB2(V-E) expressing cells did not result in a tumorigenic phenotype. In addition, p185erbB2(V-E) expressing cells displayed a reduced ability to grow in soft agar compared to the parental cell line. In contrast these transfected cells were able to grow in three-dimensional collagen gels, whereas parental cells were not. Thus, expression of erbB2(V-E) in SW 613-S cells induced multiple changes in intracellular signaling and in growth requirement phenotype, particularly in response to the extracellular environment.

Adaptor Proteins, Signal Transducing↗

Epidermal growth factor receptor signaling cascade as target for tyrphostin (RG 50864) in epithelial cells. Paradoxical effects on mitogen-activated protein kinase kinase and mitogen-activated protein kinase activities.

Tyrphostins are synthetic compounds that have been described as in vitro inhibitors of epidermal growth factor receptor (EGF-R) tyrosine kinase activity. The inhibitory effect of tyrphostins in intact cells has been shown only after prolonged treatment. However, these compounds appear to be readily incorporated, which suggests that tyrphostin acts indirectly on EGF-R. We studied the effects of a tyrphostin derivative, RG 50864, without preincubation in intact epithelial cells. We selected two human cell lines differing in degree of expression of the p185erbB2 protein, which is closely related to EGF-R. We showed that tyrphostin (RG 50864) had no effect on EGF-dependent EGF-R tyrosine phosphorylation in the parental cell line. On the contrary, it prolonged the EGF-dependent EGF-R and p185erbB2(V-E) tyrosine phosphorylation in p185erbB2(V-E)-expressing cells. Because tyrphostin has been shown to be an inhibitor of p185erbB2 and EGF-R in vitro, this finding indicates that the tyrphostin effect on p185erbB2(V-E) and EGF-R was the result of an indirect mechanism in transfected cells. Tyrphostin treatment alone led to the activation of mitogen-activated protein (MAP) kinase kinase or MAP kinase or extracellular signal-regulated kinase kinase (MEK), suggesting that one of the tyrphostin targets was upstream of MEK1. MAP kinase, however, was not activated after tyrphostin treatment. This finding indicates that tyrphostin had another target in intact cells because MEK1 activation by tyrphostin alone did not correlate with MAP kinase activation. In the two cell lines, tyrphostin modified the time course of EGF-dependent MEK and MAP kinase activation. We conclude that whereas tyrphostins were designed to inhibit EGF-R tyrosine kinase activity, under our conditions EGF-R is not a physiological target for tyrphostin, nor is one of its related protein tyrosine kinases, p185erbB2(V-E). On the contrary, our results show that tyrphostin targets are multiple, leading to complex effects on receptor signaling in these epithelial cells.

Calcium-Calmodulin-Dependent Protein Kinases↗

Upregulation of CD40 and CD40 ligand expression in IgE-associated cutaneous diseases.

In order to better understand the immunological processes connected with IgE-associated cutaneous disease, we have examined the expression of CD40 and its ligand CD40L, required for the induction of IgE synthesis in B-cells, as well as of IgE and its receptors in various dermatoses (atopic dermatitis (AD), scabies, chronic recurrent urticaria) versus normal skin, and in one dermopathic lymph node versus normal lymphatic tissue by immunohistochemistry. Compared to normal skin, cells expressing IgE, Fc epsilon RI, Fc epsilon RII, CD40, CD40L and L26 were increased in the dermis, partly also in the epidermis, from patients with AD and scabies, but not in chronic urticaria. CD40 and CD40L were detected on numerous cells in lymphatic tissue from both normal donors and a patient with AD, whereas large numbers of IgE- and Fc epsilon RI-positive cells were only found in the dermopathic lymph node from the AD patient, in contrast to very few in normal lymphatic tissue. These results with selectively increased IgE/Fc epsilon RI and associated CD40/CD40L expression in the skin of AD and scabies suggest that cutaneous tissue, in addition to dermopathic lymphatic tissue, might contribute to IgE synthesis.

Antigen Presentation↗

[Comparison of treatment outcome for non-variceal upper gastrointestinal hemorrhage using endoscopic obliteration methods and traditional methods in patients over 60].

UNLABELLED: Authors present results of therapy of patients over 60 treated because of nonvariceal upper gastrointestinal bleeding in General Surgery Department in Oświecim. In 1991-1993 47 patients underwent non-endoscopic therapy. In 1994-1996 endoscopic obliteration with 0.01% epinephrine or 1% polidocanol in patients over 60 was applied. In the group over 60 more often concomitant diseases (cardiovascular, diabetes, atherosclerosis) were noted. Similar endoscopic obliteration efficacy (90% vs. 91%) in patients aged under and over 60 was observed. Patients treated with endoscopic obliteration underwent urgent operation twice rarely then conservative management (6.7% vs. 13%). They obtained less quantity of blood (2.23 vs. 3.4 unit). Their hospital stay was shorter (9 vs. 13 days). In both kind of therapy, mortality in patients over 60 was higher then under 60, but less often in group undergone endoscopic obliteration (traditional treatment 28% vs. 6%, endoscopic 15% vs. 2%). CONCLUSIONS: 1. Reason of worse therapy results of upper gastrointestinal bleeding among patients aged over 60 was great number of concomitant diseases. 2. Efficacy of endoscopic obliteration 0.01% epinephrine and 1% polidocanol was independent from age. 3. Application of endoscopic obliteration improved results of therapy in patients aged over 60.

Aged↗

Nonlinear polarization spectroscopy in the frequency domain of light-harvesting complex II: absorption band substructure and exciton dynamics.

Spectral substructure and ultrafast excitation dynamics have been investigated in the chlorophyll (Chl) a and b Qy region of isolated plant light-harvesting complex II (LHC II). We demonstrate the feasibility of Nonlinear Polarization Spectroscopy in the frequency domain, a novel photosynthesis research laser spectroscopic technique, to determine not only ultrafast population relaxation (T1) and dephasing (T2) times, but also to reveal the complex spectral substructure in the Qy band as well as the mode(s) of absorption band broadening at room temperature (RT). The study gives further direct evidence for the existence of up to now hypothetical "Chl forms". Of particular interest is the differentiated participation of the Chl forms in energy transfer in trimeric and aggregated LHC II. Limits for T2 are given in the range of a few ten fs. Inhomogeneous broadening does not exceed the homogeneous widths of the subbands at RT. The implications of the results for the energy transfer mechanisms in the antenna are discussed.

Circular Dichroism↗

[The effect of intravenous nifedipine administration on coronary occlusion during coronary angioplasty on ischemia tolerance and collateral function].

In 29 consecutive patients (pts) coronary wedge pressure (CWP) was determined as an indicator of coronary collateral function during coronary angioplasty. Collaterals to the target vessel were angiographically detectable in 21 pts. CPW, aortic pressure (AOP), pulmonary wedge pressure (PCP), intervals to appearance of angina pectoris, surface and intracoronary ECG-changes were registered during two (n = 10) or three (n = 19) consecutive balloon dilatations. A total of 21 pts received 0.8-1.0 mg nifedipine intravenously before a second or third dilatation was performed; a control group (n = 8) received placebo. Hemodynamic parameters were reproducible for all dilatations without nifedipine. After administration of nifedipine significant changes occurred: decreases of CPW (from 34 to 29 mm Hg), AOP (from 121 to 110 mmHg), and PCP (from 12.4 to 9.4 mm Hg), and increase of ischemic tolerance time (angina pectoris) (from 35 to 56 s) (p less than 0.01). Changes in CWP and AOP showed a statistical tendency to correlate (p = 10). Thus, intravenous administration of nifedipine can improve ischemic tolerance during coronary angioplasty. Simultaneous measurement of coronary wedge pressure could not prove enhancement of collateral function as being responsible for these antiischemic effects.

Adult↗

[Pharmacokinetics and thyrotoxicity of the sodium nitroprusside metabolite thiocyanate (author's transl)].

Thiocyanate (SCN) serum levels were measured in relation to renal function and SCN-induced changes in thyroid function in 17 patients treated with sodium nitroprusside. SCN serum levels rose linearly, dose-dependent, after the distribution phase. If there was creatinine retention the rise was much steeper. Delayed SCN elimination in renal failure, previously shown experimentally, was also demonstrated clinically. Suppression of thyroid function, to a not precisely predictable extent, is likely to occur at a SCN serum level from 18 mg/l upward.

Adult↗

[Not Available].

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Historiography↗