Two types of high inductive electromagnetic stimulation and their different effects on endothelial cells.

Effects of low-frequency electromagnetic fields (LF EMF) on the activation of different tissue recovery processes have not yet been fully understood. The detailed quantification of LF EMF effects on the angiogenesis were analysed in our experiments by using cultured human and mouse endothelial cells. Two types of fields were used in the tests as follows: the LF EMF with rectangular pulses, 340-microsecond mode at a frequency of 72 Hz and peak intensity 4 mT, and the LF EMF with sinusoidal alternating waveform 5 000 Hz, amplitude-modulated by means of a special interference spectrum mode set to a frequency linear sweep from 1 to 100 Hz for 6 s and from 100 Hz to 1 Hz return also for 6 s, swing period of 12 second. Basic parameters of cultured cells measured after the LF EMF stimulus were viability and proliferation acceleration. Both types of endothelial cells (mouse and human ones) displayed significant changes in the proliferation after the application of the LF EMF under conditions of a rectangular pulse mode. Based on the results, another test of the stimulation on a more complex endothelial-fibroblast coculture model will be the future step of the investigation.

Clinical impact of PAI 1 4G/5G gene polymorphism in colorectal carcinoma patients.

Plasminogen activator ihnibitor (PAI 1) belongs to the plasminogen activator system, which is part of the metastatic cascade and significantly contributes to invasive growth and angiogenesis of malignant tumors. Its plasma level is normally low but 4G/4G homozygotes have higher concentrations of PAI 1. This genotype may be associated with worse prognosis and proximal location of colorectal cancer than 5G/5G homozygotes. In our prospective evaluation we examined plasma level PAI 1 (using photometric microplate method ELISA) pre-surgery and, subsequently, 6-8 weeks later, from 80 patients. For the PAI 1 rs1799889 -675 4G/5G polymorphism test the PCR amplification was used.Analysis of collected data was confirmed that significantly higher plasma levels of PAI 1 were found in patients before starting therapy, which decreased (p=0.004) after initiation of treatment. Patients with higher plasma level PAI 1 before (p=0.013) and after therapy (p=0.004) had significantly shorter survival. We found no relationship between polymorphisms of PAI 1 (-675 4G/5G) in relation to stage, survival or tumor location. PAI 1 is useful as a negative marker of prognosis and could be advantageous when planning adjuvant treatment of patients with colorectal carcinoma. Although opinions on the importance of polymorphisms of PAI 1 in relation to the prognosis are not uniform, it does seem that their role in the prognosis of patients with colorectal cancer is not essential.

Clinical significance of the plasminogen activator system in relation to grade of tumor and treatment response in colorectal carcinoma patients.

Urokinase (uPA) plays an essential role in the activation of plasminogen to plasmin, and together with its receptor (uPAR), tissue activator (tPA) and urokinase inhibitors (PAI 1, PAI 2, PAI 3 and protease nexin) forms the plasminogen activator system (PAS), a component of metastatic cascade importantly contributing to the invasive growth and angiogenesis of malignant tumours. In our project we examined the expression of uPA, uPAR, PAI 1 and PAI 2 in tumor tissue and we also studied the plasma levels of PAI 1 before and after the initiation of therapy in patients with colorectal carcinoma in relationship to grade of tumor and the treatment response. In our prospective evaluation we included 80 patients treated for adenocarcinoma of the colon and rectum. Analysis of collected data revealed statistically significant evidence of a relationship between the level of PAI 1 in plasma before treatment and grade of the tumor, which increases with tumor grade (p=0.025). We demonstrated that there exists a statistically significant relationship between the expression of PAI 2 (p<0.001) and uPAR (p=0.031) and grade of tumor. We also confirmed a statistically significant relationship between soluble levels of PAI 1 before treatment and therapeutic response (p=0.021). In our group of patients the expression of uPA, uPAR, PAI 1 and 2 in tumor tissue in relation to response to treatment was also assessed. Our results suggest that the greater expression of these parameters in tumor tissue is linked to a worse response to therapy. In conclusion, PAS factors help as a prognostic indicators and could also act as a predictive factor in colorectal carcinoma.

[Plasminogen activator system and its clinical significance in patients with a malignant disease]. / Plazminogen aktivátor systém a jeho klinický význam u pacientu s nádorovým onemocnením.

Urokinase (uPA) plays an essential role in the activation of plasminogen to plasmin, a serine protease participating in the activation of matrixmetaloproteinases, latent elastases, growth factors and cytokines involved in the degradation of extracellular matrix elements. Together with its receptor (uPAR), tissue activator (tPA) and urokinase inhibitors (PAI-1, PAI-2, PAI-3 and protease nexin), it forms the plasminogen activator system (PAS), a component of metastatic cascade importantly contributing to the invasive growth and angiogenesis of malignant tumours. Plasminogen activator inhibitor 1 inhibits uPA-dependent invasiveness of some cancer cell lines. The vitronectin-PAI-1 complex inhibits migration of smooth muscle cells by binding alpha(v)beta3 integrin to vitronectin. PAI-1 or its deficiency interferes with signalling pathways such as PI3K/Akt and JAK/STAT and it is included in the processes of maintaining the integrity of the endothelial cells and thereby regulation of cell death. PAI-1 affects apoptosis by reducing cell adhesion and functioning of intracellular signalling pathways. The individual components of PAS undoubtedly play an important role in angiogenesis and metastasising of malignant tumours. In the near future, results of published studies with various types of cancer could be reflected in diagnostic and therapeutic algorithms and, at the same time, could serve as the goal for targeted therapies.

[Determination of the activity of luteinizing hormone in PMSG preparations using testosterone synthesis in vitro]. / Stanovení aktivity luteinizacního hormonu v preparátech PMSG podle syntézy testosteronu in vitro.

The activity of luteinizing hormone (LH) in serum gonadotropin (PMSG) was studied by a bioassay in vitro, using the cells of mouse testes prepared by the combination of enzymatic and mechanical diffociation. The activity of individual preparations was calculated according to the amount of produced testosterone measured by radioimmunoassay. Comparison with the 2nd international standard of PMSG (WHO) showed that in the PMSG substandard (Dessau) the activity of LH was twice lower, and in three charges of the commercial PMSG preparation (Bioveta, Ivanovice in Haná) four-and-a-half times lower than declared. No great variability of LH activity was observed among the charges of the commercial PMSG preparation.

[The effect of different cell concentrations and incubation time of testicular tissue testosterone synthesis]. / Vliv ruzných bunecných koncentrací a doby inkubace varletní tkáne na syntézu testosteronu.

The cells of the testes of mice were enzymatically separated and the amount of Leydig cells was determined in different stages of dispersion and after washing the separated tissue. This amount ranged from 2 to 9%. The largest quantity of Leydig cells (9%) was obtained when separated tubuli were washed. The response of cell suspensions at seven concentrations stimulated by human chorion gonadotrophin (HCG) was tested. Cell suspension concentrations from 5.3 x 10(5) to 1.4 x 10(6) cells/ml were found to be satisfactory. After two to three hours of incubation, the values of testosterone could be used for the determination of the activity of gonadotropic preparations.

[Determination of the gonadotropin activity of biological preparations by testosterone synthesis in mouse testes in vitro]. / Stanovení gonadotropní aktivity biopreparátu podle syntézy testosteronu mysími varlaty in vitro.

The ability of mouse testicular tissue to respond to stimulation by a commercial preparation of human choriongonadotropin (Praedyn Spofa) was tested in vitro, using whole testes, fragments and cell suspension. In each experiment seven gonadotropin concentrations ranging from 0.06 to 50 i. u. per litre were tested, and the results were compared with the group of control samples without gonadotropin. The incubation of whole testes with gonadotropin of different concentrations was performed in five vessels, each containing one testis; the fragments were incubated in four vessels, each containing four fragments from different testes, and the cell suspension in triplicates containing 1 . 10(6) cells per ml. All three types of tissue reacted to the increasing concentrations of stimulating agent by the increasing production of testosterone, the concentration of which was determined by radioimmunoassay. For the suppression of the individuality and for achieving a uniform response of the individual samples with each stimulation, the cell suspension of testes from several animals appears to be most suitable.