[The influence of physical load on the propriate immune and physiological parameters.] / Vliv fyzické záteze na vybrané imunitní a fyziologické parametry.

Chronic fatigue syndrome is a disease detected in recent 15 or 20 years. Overtrained athletes, people living in stress, the ones with disturbed immunity or people suffering from some of the infectious diseases are the most threatened ones. During ultra-long-distance run, human immune, physiological a biochemical parameters drift of their physiological ranges. The values could increase or decrease. The samples of serum of ultramarathon runners, who took part in the National Ultramarathon Mastership, were collected and measured before and after the race. The parameters include IgA, IgM, IgG and C3 part of complement. Statistically important increases in IgA and IgG concentrations after the race were observed. The changes of concentrations of IgM and C3 part of complement was not statistically important. IgG is responsible for the activation of complement, secondary immune reactions and the neutralization of bacterial toxins. IgA in the role of muckal imunoglobulin helps immune cells to swallow heterogenous particles, germs and toxins. Our immune syst6m is more threatened by heterogenous infectious diseases and even the chronic fatigue syndrome.

Development of a novel enzyme-linked immunosorbent assay (ELISA) for measurement of serum CTRP1: a pilot study: measurement of serum CTRP1 in healthy donors and patients with metabolic syndrome.

BACKGROUND: Complement C1q tumor necrosis factor-related protein 1 (CTRP1), a recently identified adipokine, was found to stimulate aldosterone production. Obesity and metabolic syndrome are frequently associated with elevated levels of aldosterone. Therefore, it would be interesting to investigate whether the secretion of CTRP1 in human serum is associated with obesity as well as with hypertension. AIM: This study evaluated serum CTRP1 concentrations in healthy individuals and patients with metabolic syndrome. METHODS: Serum samples from 61 healthy individuals and 46 patients with metabolic syndrome were measured for CTRP1 by enzyme-linked immunosorbent assay (ELISA). RESULTS: Correlation analyses showed that serum CTRP1 in healthy individuals did not correlate with BMI, leptin, TG, HDL-CH, and LDL-CH; however, in patients with metabolic syndrome, CTRP1 correlated with glucose, HbA1c and BMI. CTRP1 level was significantly higher in subjects with metabolic syndrome compared to healthy subjects. DISCUSSION: Our results support the hypothesis that adipokine CTRP1 is associated with metabolic syndrome and obesity compared to healthy individuals.

Activity of the tick Ixodes ricinus monitored in a suburban park in Brno, Czech Republic, in association with the evaluation of selected repellents.

The ever-increasing number of Lyme borreliosis patients led us to consider more effective procedures for disease prevention. The aim of our study was to monitor the annual activity and infectivity of Ixodes ricinus ticks in the Pisárky region, City of Brno, CR, and to test the responses of the locally-captured ticks to selected repellents. The result of regular one-hour-per-week monitoring in 2011 was the collection of ticks that directly reflected the highest number of Lyme disease patients (4,835) detected throughout the period of recording in the Czech Republic. The ticks were examined for spirochaetes by dark field microscopy. The positive samples were identified by PCR analysis, confirming that 76% of these were infected with Borrelia burgdorferi sensu lato. Ticks were most abundant in May and June, with August having the highest risk for spirochaetal infection. Tick activity was statistically correlated with temperature. The moving-object-bioassay was used to study repellent efficiency on the Ixodes ricinus nymphs captured in the above-mentioned suburban park. Five selected commercial repellents based on DEET (N, N-diethyl-3-thylbenzamide) showed statistically different effects on the non-repellent control group.

Influence of environmental factors on the occurrence of Ixodes ricinus ticks in the urban locality of Brno - Pisárky, Czech Republic.

The occurrence of Ixodes ricinus ticks was observed in the suburban locality of Brno-Pisárky (South Moravia, Czech Republic) from March to November, 1996 to 2002. A total of 2,813 ticks was collected. Statistical tests divided the activity of ticks into three periods during the year. The curve of seasonality had two peaks with a maximum in May and August, with a significantly larger number of specimens collected during this period compared to other months. The abundance of ticks during Spring and Autumn months was comparable. All developmental stadia of ticks were found in this locality. The number of larvae, males, and females was not significantly different, but the occurrence of nymphs was significantly greater. Except for the year 2000, there were no statistically significant differences in tick abundance. Tick activity was not dependent on humidity but did vary directly with temperature. This relation had a linear character and could be described by the equation y = 8.3 + 1.8x.

Serological confirmation of Borrelia burgdorferi infection in dogs in the Czech Republic.

From the epidemiological point of view, dogs are very important since they are considered a suitable indicator of the spread of human borreliosis. Serum samples obtained from healthy, asymptomatic military dogs from 12 different areas in the Czech Republic were examined for IgG antibodies to Borrelia burgdorferi sensu lato (s.l.). The total of 399 serum samples were tested by a whole-cell ELISA. Specific antibodies to Borrelia burgdorferi s.l. were detected in 26 cases (6.5%). In different localities, the seroprevalence varied from 0.0% to 28.6%. Two local isolated strains Br-75 (Borrelia afzelii) and Br-97 (Borrelia garinii) were used as antigens. A total of 22 (5.5%) were positive for antibodies to Borrelia afzelii and 19 (4.8%) were positive for antibodies to Borrelia garinii. Fifteen cases were positive for both antibodies. A significantly higher seroprevalence was found in younger dogs (1-3 years) than in older ones (p < 0.05). An analysis of seroprevalence by months of sampling showed no significant difference (p > 0.05).

Presence of Borrelia burgdorferi sensu lato in mites parasitizing small rodents.

Ticks (especially those in the family Ixodidae) are the primary vectors of the infectious agent Borrelia burgdorferi sensu lato, which causes Lyme disease. To determine the potential role of mites as vectors of B. burgdorferi sensu lato, mites were collected from wild rodents in the Bazantula region of North Moravia (Czech Republic). These samples were examined for the presence of Borreliae by using DFM and PCR methods. Mites positive for the presence of DNA of B. burgdorferi sensu lato were determined as members of the families Haemogamasidae and Parasitidae. One sample from a mite of genus Haemogamasus was successfully isolated, and the specimen was confirmed as B. afzelii by using PCR-RFLP and by gradient SDS-PAGE. This suggests the possible participation of gamasid mites in borrelial circulation in nature and also points to the utility of further such studies to identify potential vectors (other than ticks) of the spirochete.

DNA interactions of antitumor trans-[PtCl2(NH3)(quinoline)].

Recent observations that several trans-platinum complexes exhibit antitumor activity including activity in cisplatin-resistant tumor cells, violates the classical structure/activity relationships of platinum(II) complexes. According to these relationships, only bifunctional platinum(II) complexes with cis-oriented leaving ligands should be therapeutically active. In order to contribute to the understanding of mechanisms underlying the antitumor activity of these new trans-platinum analogs, various biochemical and biophysical methods as well as molecular modeling techniques were employed to study the modifications of DNA by antitumor trans-[PtCl2(NH3)(quinoline)]. The results indicated that trans-[PtCl2(NH3)(quinoline)] coordinated monofunctionally to DNA with a similar rate as transplatin. The overall rate of the rearrangement to bifunctional adducts was also similar to that observed in the case of DNA modification by transplatin, i.e. it was relatively slow (after 48 h approximately 34% adducts remained monofunctional). In contrast to transplatin, however, trans-[PtCl2(NH3)(quinoline)] formed considerably more interstrand cross-links after 48 h (approximately 30%) with a much shorter half-time (approximately 5 h) (approximately 12% for transplatin, t1/2 > 11 h). The results also suggested that the quinoline ligand in all or in a significant fraction of DNA adducts of trans-[PtCl2(NH3)(quinoline)], in which platinum is coordinated to base residues, was well positioned to interact with the duplex. The adducts of trans-[PtCl2(NH3)(quinoline)] terminated in vitro RNA synthesis preferentially at guanine residues. Surprisingly, the type and extent of conformational alterations induced in DNA indicates that trans-[PtCl2(NH3)(quinoline)] behaves in some respects like cisplatin, as indicated by the fact that trans-[PtCl2(NH3)(quinoline)]-modified DNA is recognized by cisplatin-specific antibodies. Models for both monofunctional adducts and bifunctional interstrand cross-links are proposed. Computer-generated AMBER models show that the combination of monofunctional covalent binding and a stacking interaction between quinoline and the DNA bases can produce a kink in the duplex which is strongly suggestive of the directed bend produced by the major cisplatin-DNA adduct (1,2 intrastrand cross-link). Unique DNA adducts of this type formed by trans-[PtCl2(NH3)(quinoline)] may contribute to the antitumor efficacy of this agent.

DNA interactions of a novel platinum drug, cis-[PtCl(NH3)2(N7-acyclovir)]+.

We synthesized a novel platinum drug, cis-[PtCl(NH3)2(N7-ACV)]+, in which ACV is the antiviral drug acyclovir [a deoxyriboguanosine analogue, 9-(2-hydroxyethoxymethyl)guanine]. This new compound exhibits antiviral efficacy in vitro and exhibits an antitumor activity profile different from that of cisplatin [Metal-Based Drugs 2:249-256 (1995)]. To contribute to understanding the mechanisms underlying biological activity of this new compound, we studied modifications of natural and synthetic DNAs in cell-free media by cis-[PtCl(NH3)2(N7-ACV)]+ by various biochemical and biophysical methods. The results indicated that the major DNA adduct of cis-[PtCl(NH3)2(N7-ACV)]+ was a stable monofunctional adduct at guanine residues. In contrast to DNA adducts of other monodentate and clinically ineffective platinum(II) compounds, the adducts of cis-[PtCl(NH3)2(N7-ACV)]+ terminated in vitro DNA and RNA synthesis. In addition, although DNA adducts of cis-[PtCl(NH3)2(N7-ACV)]+ and cisplatin were different, some properties of DNA modified by either compound were qualitatively similar. Such similarities were not noticed if DNA modifications by other ineffective monofunctional platinum(II) complexes were investigated. Thus, the DNA binding mode of monofunctional cis-[PtCl(NH3)2(N7-ACV)]+ was different from that of other monofunctional but ineffective platinum(II) complexes. It has been suggested that the unique capability of cis-[PtCl(NH3)2(N7-ACV)]+ to modify DNA may be relevant to a distinct antitumor efficiency of this novel drug in comparison with cisplatin. It also has been suggested that at least some aspects of DNA interactions of cis-[PtCl(NH3)2(ACV)]+ revealed in the current study could be exploited in the search for and development of new antiviral platinum complexes containing, as a part of the coordination sphere, antiviral nucleosides.

DNA modifications by antitumor trans-[PtCl2(E-iminoether)2].

Recent findings that an analogue of clinically ineffective transplatin, trans-[PtCl2(E-iminoether)2], exhibits antitumor activity has helped reevaluation of the empirical structure-antitumor activity relationship generally accepted for platinum(II) complexes. According to this relationship, only the cis geometry of leaving ligands in the bifunctional platinum(II) complexes, should be therapeutically active. Global modifications of natural DNAs in cell-free media by trans-[PtCl2(E-iminoether)2] were studied through various molecular biophysical methods and compared with modifications by cis-[PtCl2(E-iminoether)2], transplatin, cisplatin, and monofunctional chlorodiethylenetriamineplatinum(II) chloride. Thus, the results of this study have extended our recent finding, indicating that the prevalent lesion occurring in double-helical DNA on its modification by trans-[PtCl2(E-iminoether)2] is a monofunctional adduct at guanine residues. The modification by trans-[PtCl2(E-iminoether)2] has been found to induce local distortions in DNA, which have a character differing fundamentally from those induced by both clinically ineffective or antitumor platinum complexes tested in this study. The different character of alterations induced in DNA by the adducts of trans-[PtCl2(E-iminoether)2] and transplatin has been suggested to be relevant to the unexpected observation that the new complex with leaving chloride groups in trans position exhibits antitumor efficacy. In addition, the results support the idea that platinum drugs that bind to DNA in a manner fundamentally different from that of cisplatin can exhibit altered biological properties, including differing spectra and intensities of antitumor activity.

DNA interactions of bifunctional dinuclear platinum(II) antitumor agents.

Modifications of natural DNA in a cell-free medium by dinuclear bisplatinum complexes with equivalent coordination spheres, represented by the general formula [¿trans-PtCl(NH3)2¿2(H2N-R-NH2)]2+, where R is a propane or hexane, were studied by various methods of biochemical analysis or molecular biophysics. These methods include binding studies by means of differential-pulse polarography, measurements of melting curves with the aid of absorption spectrophotometry, measurements of CD spectra, ELISA with specific antibodies that recognize DNA modified by platinum complexes, interstrand cross-linking assay employing gel electrophoresis under denaturing conditions and mapping of DNA adducts by means of transcription assays. The results indicated that the major adduct of [¿trans-PtCl(NH3)2¿2(H2N-R-NH2)]2+ in DNA was an interstrand cross-link which was formed with a relatively short half-time (approximately 1 h). At least some types of these interstrand cross-links induced local denaturational changes in the DNA. The results of analyses of interactions of [¿trans-PtCl(NH3)2¿2(H2N-R-NH2)]2+ with linear DNA at relatively higher levels of the modification could be interpreted to mean that these dinuclear platinum complexes were also capable of intrastrand-cross-link formation between adjacent base residues in DNA. However, these intrastrand adducts of [¿trans-PtCl(NH3)2¿2(H2N-R-NH2)]2+ distorted DNA conformation in a way different from the DNA intrastrand adducts of cisplatin. In addition, the DNA adducts of the dinuclear platinum complexes inhibited DNA transcription in vitro. The length of the aliphatic linker chain affected the DNA-binding mode of [¿trans-PtCl(NH3)2¿2(H2N-R-NH2)]2+ and the resulting conformational changes in DNA. The extensive analysis of DNA interactions with [¿trans-PtCl(NH3)2¿2(H2N-R-NH2)]2+ described in this communication has provided further experimental support for previous suggestions [Farrell, N. (1991) in Platinum and other metal coordination compounds in cancer chemotherapy (Howell, S. B., ed.) pp. 81-91, Plenum Press, New York] that the binding of the dinuclear platinum complexes modifies DNA in a way that is different from the modification by antitumor cisplatin. Thus, the results of this work are consistent with the hypothesis that platinum drugs that bind to DNA in a manner fundamentally different from that of cisplatin can exhibit altered biological properties, including a different spectrum and intensity of antitumor activity.