High inductive magnetic stimuli and their effects on mesenchymal stromal cells, dendritic cells, and fibroblasts.

Effects of low-frequency electromagnetic fields (LF EMF) on the activation of different tissue recovery processes have already been fully understood. Preliminary recent data demonstrated that a special case of sinusoidal electromagnetic fields, known as amplitude-modulated currents (AMC) could have a potential to accelerate the cell metabolism or cell migration. An AMC generator was designed to generate sinusoidal induced electric currents with the amplitude modulation and the harmonic carrier frequency of 5,000 Hz was modulated by frequencies of 1 to 100 Hz. The magnetic field peak was 6 mT, electric field intensity 2 V/m and the current density of induced electrical currents was approximately 1 A/m(2). The coil of the generator was adapted to easy handling and safe integration into the shelf of the CO(2) incubator. The shelf with the coil was prepared for the introduction of cells in standard plastic in vitro chambers. The tests focused on cells with migratory capacity after injury or during immunological processes and thus, mesenchymal stromal cells (MSC), dendritic cells (DC), and fibroblasts were chosen. The tests involved exposures of the cells to LF EMF (180 min/day) every day, for a period of three days, before examining them for cell death, morphology changes, and CD markers. The samples were tested by using MTT assay and the effects on the intracellular concentration of reactive oxygen species were quantified. The cell migration was finally measured with the help of the transwell migration assay. None of the cell types showed any decrease in the cell viability after the LF EMF application and the cells displayed minimum changes in reactive oxygen species. Functional changes (acceleration of cell migration) after AMC exposure were statistically significant for the MSC samples only. The acceleration of MSCs is associated with the production of MMP by these cells. The EMF has a potential to be a safe, clinically applicable selective activator of MSC homing, MSC paracrine production, and subsequent regeneration processes.

Persistence of genetic damage in lymphocytes from former uranium miners.

Lymphocytes from former uranium miners who finished work underground one or more decades ago were analysed with respect to possibly persisting genetic damage induced by their radiation exposure. A modified micronucleus-centromere test was used which determined the frequency of micronucleus-containing binucleate cells after cytochalasin B treatment and the percentage of centromere-free micronuclei, assessed with the help of immunofluorescence labeling of centromere protein B. Whereas the overall frequency of micronucleus-containing cells was not significantly elevated above the level found in a control group, former miners showed a greater percentage of centromere-free micronuclei, i.e. micronuclei containing only acentric fragments. Our results are in excellent agreement with those of an earlier uranium miner study and lend support to the assumption that genetic damage from alpha radiation can persist for many years after exposure, possibly due to genomic instability. The frequency of micronucleus-containing cells, but not the percentage of centromere-free micronuclei, significantly increased with time since last exposure in the mines. This can be attributed, at least in part, to the fact that miners who have finished working underground longer ago tend to be older, and there is an increase of the frequency of micronucleus-containing cells with age.

Occupational exposure control: the problem of quantities in radiation protection.

The paper explores the quantities and units used in radiation protection with special emphasis on their applications in occupational exposure control. An overview of the current situation reveals that there seem to be too many different quantities associated with the same unit. Some of these quantities are defined in a quite complicated manner and, therefore, may cause some confusion in their interpretation and practical use in the field. Some suggestions towards the simplification of the present system are also proposed.

Assessment of the total effective dose of miners in the underground Rozná Uranium Mine in the Czech Republic during the period 2004-2009.

The paper discusses the situation in the Czech Republic regarding past and present uranium mining activities with emphasis on the evaluation of the exposure of underground miners in the Rozná Uranium Mine, which is currently the only active mine in the country and practically in the entire European Union. The total effective dose has been summarised taking into account all three major components, namely radon short-lived decay products, long-lived alpha emitters in ore dust and penetrating external gamma radiation. The average and maximum values of the effective dose as well as the collective effective dose of underground miners are also presented. The purpose of the paper is to document the miners' exposures during a period of 6 years in a uranium mine where conditions including the ore grade and methods of mining showed recently some changes that may affect the individual components of the total effective dose.

Effectiveness of low-level laser therapy in temporomandibular joint disorders: a placebo-controlled study.

OBJECTIVE: Low-level laser therapy (LLLT) treatment for pain caused by temporomandibular joint disorders (TMD) was investigated in a controlled study comparing applied energy density, subgroups of TMD, and duration of disorders. BACKGROUND DATA: Although LLLT is a physical therapy used in the treatment of musculoskeletal disorders, there is little evidence for its effectiveness in the treatment of TMD. METHODS: The study group of 61 patients was treated with 10 J/cm(2) or 15 J/cm(2), and the control group of 19 patients was treated with 0.1 J/cm(2). LLLT was performed by a GaAlAs diode laser with output of 400 mW emitting radiation wavelength of 830 nm in 10 sessions. The probe with aperture 0.2 cm(2) was placed over the painful muscle spots in the patients with myofascial pain. In patients with TMD arthralgia the probe was placed behind, in front of, and above the mandibular condyle, and into the meatus acusticus externus. Changes in pain were evaluated by self-administered questionnaire. RESULTS: Application of 10 J/cm(2) or 15 J/cm(2) was significantly more effective in reducing pain compared to placebo, but there were no significant differences between the energy densities used in the study group and between patients with myofascial pain and temporomandibular joint arthralgia. Results were marked in those with chronic pain. CONCLUSION: The results suggest that LLLT (application of 10 J/cm(2) and 15 J/cm(2)) can be considered as a useful method for the treatment of TMD-related pain, especially long lasting pain.

Arthralgia of the temporomandibular joint and low-level laser therapy.

OBJECTIVE: This case report describes the treatment of a patient with arthralgia of the temporomandibular joint (TMJ) caused by disc displacement. BACKGROUND DATA: The goal of the treatment of TMJ arthralgia is to decrease pain by promotion of the musculoskeletal system's natural healing ability. METHODS: This report describes the complex treatment of TMJ arthralgia. Low-level laser therapy (LLLT) was chosen for its antiinflammatory and analgesic effects. Laser therapy was carried out using the GaAlAs diode laser with an output power of 400 mW, emitting radiation with a wavelength of 830 nm, and having energy density of 15 J/cm2; the laser radiation was applied by contact mode on four targeted spots in 10 sessions. Physiotherapy was recommended to this patient to prevent the injury of intraarticular tissue caused by incorrect movement during opening of the mouth. Splint stabilization and prosthetic treatment were used to reduce overloading of the TMJ, resulting from unstable occlusion and to help repositioning of the dislocated disc. RESULTS: Five applications of LLLT led to decrease of pain in the area of the TMJ on the Visual Analog Scale, from 20 to 5 mm. The anti-inflammatory effect of the laser was confirmed by thermographic examination. Before treatment, the temperature differences between the areas of the normal TMJ and TMJ with arthralgia was higher than 0.5 degrees C. However, at the conclusion of LLLT, temperatures in the areas surrounding the TMJ were equalized. CONCLUSION: This study showed the effectiveness of complex non-invasive treatment in patients with arthralgia of the TMJ. The analgesic and anti-inflammatory effects of LLLT were confirmed by infrared thermography.

Cell analysis with the new Leipzig high-energy ion nanoprobe.

The high-energy ion nanoprobe LIPSION at the University of Leipzig has been in operation since 1998. The ultrastable, 3.5 MV SINLETRON accelerator supplies the H+ or He+ ion beam. A magnetic scanning system moves the focused beam across the sample. At present, a resolution of 41 +/- 4 nm in the low current mode and 300 nm at 5 pA can be achieved. The experimental chamber is equipped with electron-, energy dispersive X-ray-, and particle detectors. They can be used simultaneously to analyse the sample by means of PIXE (particle induced X-ray emission), RBS (Rutherford backscattering), and in the case of thin sections or monolayer samples STIM (scanning transmission ion microscopy). A goniometer allows the application of channeling measurements in single crystals in combination with these methods. In contrast to previous publication describing microbeam facility at LIPSION, the current biomedical research has concentrated on microscopy and tomography on chondrocytes in pig cartilages and fixed single endothelial cells (HUVEC). For the irradiation of single living cells, an external beam facility with irradiation platform, fast beamgate and mini-Petri dishes is under construction.

The alveolar septal thickness and type II pneumocytes number in irradiated lungs, time expression and the effect of pentoxifylline.

PURPOSE: We studied the relationship between type II pneumocytes number and alveolar septal thickness during different time after sublethal whole-thorax irradiation of rats and we investigated the influence of pentoxifylline (TNF-alpha inhibitor). MATERIALS AND METHODS: Wistar rats were exposed to 15 Gy thoracic irradiation and pentoxifylline (35 mg/kg) twice a week. Lungs were examined histologically and immunohistochemically at intervals ranging from 1-12 weeks and alveolar septal thickness, number of type II pneumocytes (identified by immunoreactivity for cytokeratin 18), and neutrophile granulocytes were counted. RESULTS: Significant increase of alveolar septal thickness and type II pneumocytes depletion 3 weeks after irradiation were found. Correlation of these markers was r = -0.759. Pentoxifylline significantly inhibits increased alveolar septal thickness without the influence on type II pneumocytes number. Neutrophil penetration started 5 weeks after irradiation in non-treated animals, 8 weeks after irradiation in PTX-treated rats. CONCLUSIONS: We suggest that pneumocytes depletion is linked to increased vascular permeability, and pentoxifylline therapy does not influence on pneumocytes kinetics after irradiation.

Apoptosis and bcl-2 expression in irradiated lungs and the effect of pentoxifylline.

We measured number of bcl-2, apoptotic, neutrophil, and surfactant apoprotein D (SP-D) positive cells in irradiated rat lungs during different time points after the sublethal whole-thorax irradiation of rats. We also investigated the influence of pentoxifylline (PTX) therapy on these markers. Wistar rats were given 15 Gy thoracic irradiation and PTX (35 mg/kg) twice a week. Animals were examined histologically and imunohistochemically at intervals from 1-12 weeks. In non-treated rats compared with treated rats, bcl-2 expression was significantly inhibited from 4 weeks after irradiation. A higher apoptosis presence in non-treated rats from 4 weeks was found and apoptosis development in PTX-treated animals was delayed and started 8 weeks after irradiation. Similar differences were measured during neutrophil granulocytes examination. Neutrophil penetration in non-treated rats was found 5 weeks after irradiation in contrast to the RP onset of PTX-treated animals 8 weeks after irradiation. The number of SP-D positive cells in non-treated rats observed until 5 weeks after irradiation was higher than in the control group. PTX-treated animals expressed higher number of SP-D positive cells during the whole experiment than the control group. We suggest that apoptosis is linked to neutrophil granulocyte actions during the RP onset and that PTX-therapy causes diminished inflammation development.

Our experiences with combination of enzyme therapy and laser therapy in enthesopathies.

The present communication brings a contribution to the confrontation of two treatment methods in enthesopathies (painful overload syndromes). In group A, 26 patients were treated by a combination of systemic enzyme therapy (SET) and non-invasive laser therapy. In group B, 13 persons were treated only by therapeutic laser. The average age in group A and B was of 39.3 and 41.8 years, respectively. The representation of men and women in the two groups was almost identical. The intensity of the pain prior to the treatment and after the treatment, specified by each patient with the help of a ten-grade scale, was evaluated by the variance analysis. The combined treatment by the SET and therapeutic laser was statistically significantly more efficient (p = 0.02). On the other hand, there was also a significant difference between the two groups in baseline values of average pain intensities. These values were of 7.0 and 5.2 in groups A and B, respectively. The average value of the pain intensity in all the 39 patients was of 6.6 prior to the treatment and it was reduced to 3.5 after the treatment.

Antitumor action of bovine seminal ribonuclease.

Unlike the bovine pancreatic ribonuclease (RNAase A), bovine seminal ribonuclease (BS RNAase) displays various biological activities including antitumor cytotoxicity. To learn more about its antitumor activity, we investigated BS RNAase effect on athymic nude mice bearing various tumors. BS RNAase (250 micrograms per mouse per day) was administered to the mice with prostate carcinoma for three weeks by three different routes (intraperitoneally--i.p., subcutaneously--s.c., and intratumorally-i.t.). Administration i.p. was ineffective, while s.c. administration reduced significantly size of tumors and i.t. administration abolished half of the tumors in treated mice. The i.t. administration of BS RNase to nude mice bearing melanoma showed even better results. Eighty % of mice were without tumors and in the other mice the tumors were significantly diminished. The best antitumor effect was obtained in case of seminoma. All mice bearing this tumor were cured after ten doses of BS RNAase.

Antitumor action of bovine seminal ribonuclease. Cytostatic effect on human melanoma and mouse seminoma.

This paper reports on the antitumor activity of BS RNase on human melanoma and mouse seminoma. Human melanoma cells established in culture were extremely susceptible to BS RNase, administered in concentrations ranging from 1-100 microg/ml. Concentrations of BS RNase over 10 microg/ml caused complete inhibition of cell growth. Bovine pancreatic ribonuclease (RNase A), a prototype of the ribonuclease superfamily, did not exert any effect under these conditions. Based on our previous results, athymic mice bearing human melanoma or mouse seminoma were treated with intratumoral administration of BS RNase (12.5 mg/kg b.w.). This dose was injected for five consecutive days excluding weekends. The intratumoral administration of BS RNase to nude mice bearing human melanoma showed a significant antitumor effect. There were no tumors seen in eighty percent of mice treated for three weeks, and tumors in the other mice diminished significantly. After some delay the tumors started to regrow. Prolonging of the treatment to five weeks had a similar effect. The effect of BS RNase on mouse seminoma was well pronounced. Five to seven doses of BS RNase were sufficient to eliminate tumors in all treated mice. However, as in the previous experiment, the growth of tumor tissue later reappeared.

Our experience in brachytherapy of certain non-cancerous diseases.

Brachytherapy of non-cancerous diseases has been used from the beginning of our century. The analgetic and antiinflammatory effects of gamma rays of the radionuclide Ra-226 has been used in the health resort in Jáchymov, located in Ore Mountains, Northern Bohemia since about 1911 in the therapy of patients hospitalized here with rheumatic diseases of joints, tendons and muscles, including the therapy of radical syndromes. The conditions for the therapy by this method are the age of above 42 years, normal blood count and certain other parameters commonly used in the non-cancer brachytherapy. The radiation dose depends on the degree of the involvement. The local single dose in our patients was of 1.8 to 3.5 Gy with an exposure time of 6 hours. In our work, we considered the degree of the damage to the skin on the irradiation site, after single dose, immediately after the application, as well as after the repeated irradiation in the course of several years. In persons followed, we also provided the examination of the blood count before and after irradiation. The results obtained demonstrated that the brachytherapy used in the health resort Jáchymov is safe and that it exerts no adverse effects on the patients submitted to it.

Mechanisms of biostimulating effects of therapeutic laser in vivo. Review article.

The beneficial biostimulating effects induced by therapeutic laser in involved tissues was checked by a number of experimental as well as clinical works. Most of them particularly pay their attention to the description of the basic disease; there is, however, considerable imprecision in the technical characterization of the laser appliances used. Only the wave length and the power of the source are typically characterized. Further principal data are, however, missing, such as the power density, energy density, etc. These communications also suggest that their authors are not always completely familiarized with the mechanism of the stimulating action low-power laser beams in tissues. Thus, the purpose of the present communication is to summarize the existing knowledge concerning this type of treatment with considering possibilities of its use in clinical practice.

[Possible therapeutic applications of pulsed magnetic fields]. / Moznosti lécebného vyuzití pulzního magnetického pole.

Magnetotherapy is a relatively new, nowadays however, relatively widespread method in several medical disciplines. The mechanism proper of the favourable action of the pulsed magnetic field on the living organism is not quite clear so far, clinical investigations revealed, however, a favourable anti-inflammatory, angioedematous and analgesic therapeutic effect. The authors sought an optimal frequency of the pulsed magnetic field with regard to the character of the disease. They focused attention above all on treatment of acute and chronic inflammatory conditions of the locomotor apparatus, ischaemia of the blood vessels of the lower extremities, dyspeptic syndrome, lactation mastitis and other diseases. One therapeutic cycle lasted 20 minutes, the mean number of cycles varied between 5.8 and 7.7. A regression of complaints was recorded as a rule after 2-3 sessions. The optimal frequency of the pulsed magnetic field seems to be a value between 10.0 and 25.0 Hz. It is useful in particular in severe conditions to repeat the therapeutic cycle after 2-3 months. The advantage of this therapeutic method is the minimal number of contraindications.

Changes of certain biochemical values during whole-body exposures of rats to doses of 50, 100 and 250 Gy.

In the course of the postirradiation syndrome, changes of concentrations of certain components and of activities of certain enzymes occur in the blood serum. The data available in the literature concern doses lower than 10 Gy. In the present work, changes were studied of concentrations of mineral components, total proteins, glucose, creatinine and bilirubin and activities of liver transaminases, acid prostatic and alkaline phosphatases up to 24 hours after the whole body exposures of rats to doses of 50, 100 and 250 Gy of ionizing radiation. The changes observed--enhanced kalemia and activity of aspartate aminotransferase, particularly after the highest radiation dose used, and changes of the glycaemia (early increase with a subsequent considerable decrease)--support the assumptions based on the former results known from the literature.

Effect of continuous, long-term administrations of acetylsalicylic acid on hematological and hemocoagulation changes in the rat.

Acetylsalicylic acid is currently being widely used either itself or in combination with the other substances for the prevention of thromboembolic events. Its effect is particularly given by the inhibition of the cyclooxygenase enzyme participating in the metabolism of eicosanoids. The recommended doses of acetylsalicylic acid as well as frequency of administrations are very different. The present work is aimed at estimating the effect of the continuous administration of acetylsalicylic acid for 28 days on hematological and hemocoagulation changes in the rat. The experimental results suggest that long-term administrations of acetylsalicylic acid should be used cautiously in human medicine. They signalize the need of long-term, aimed clinical studies of hematological parameters of patients preventively treated with acetylsalicylic acid, also with respect to the producer of this substance. It is also impossible to neglect regular controls of necessary parameters for the estimation of the efficiency of the given therapy in each patient with respect to individual reactions. The dosage should also be adjusted with respect to the age, sex and individual sensitivity of the organism.

Influence of cloprostenol-15-glycal on some hematologic values.

Cloprostenol-15-glycal is an analog of natural prostaglandin F2 alpha. It is prepared fully synthetically and is a basic substance of the veterinary medicament of firm name Glystrofan (chem. conc. Spolana, Neratovice, CSR). The effect of this substance was studied after intravenous application in double concentration on the blood cell count in experiment on rats. A significant decrease of erythrocyte count was observed in late sampling intervals especially after higher dose. A temporary elevation of leucocyte count occurred in white blood cell series. There was lymphocyte count elevation, while neutrophil count decreased. In the course of experiment a decrease of thrombocyte count occurred, especially after higher dose. Characteristic of described changes corresponds to that, which was observed by prostaglandins of series E. Changes in perifery blood cell count can be in agreement with the effect of cloprostenol-15-glycal on the production or influence of growth factors and on the proliferation of pluripotent hematogenic cells respectively.

Changes of 6-keto PGF1 alpha concentration in plasma and vessel wall and TxB2 in plasma of whole-body irradiated rats in the early stage of irradiation.

The aim of this work was to monitor concentrations of prostanoids affecting platelet aggregation, namely prostacyclin and thromboxane A2, in the plasma and prostacyclin in the endothelium of vessel wall after whole-body irradiation of rats by the doses 5, 10, 50, 100 and 250 Gy (source 60Co) in the early stage of irradiation up to 24 hours. Stable metabolites of the prostanoids - 6-keto PGF1 alpha and TxB2 were determined by a RIA technique. The 6-keto PGF1 alpha plasma concentration increased after irradiation by the 50 Gy dose between 3rd and 24th hours and at all sampling times after the 100 and 250 Gy doses. A rise in prostanoid concentrations was noted after low doses of irradiation in the first sampling intervals only. The TxB2 level increased after the same doses (5 and 10 Gy) up to the 6th hour and then decreased. The thromboxane concentration increased after 50 Gy at the 90th minute, 12th and 24th hours, after 100 Gy at the 24th hour and after 250 Gy at the 90th minute, 3rd and 24th hours.

[Our initial experience with Czechoslovak-made indium-111-oxine used for leukocyte labeling]. / Nase první zkusenosti s radiofarmakem oxinát inditý-111 Ceskoslovenské výroby pri znacení leukocytu.

Labelling of leucocytes with indium oxinate is currently used abroad. Labelled leucocytes are used mainly for the detection of inflammatory processes. The aim of the work was to test the binding capacity of the radiopharmaceutical preparation indium111 oxinate prepared in the Nuclear Research Institute of the Czechoslovak Academy of Science in Rez, its effect on the survival of leucocytes and its sterility. The authors' aim was also to use during separation of leucocytes Czechoslovak preparations to use this technique on a large scale in Czechoslovakia. The assembled results indicate a good quality of the used pharmaceutical preparation; the assessed values of the percentage of activity linked to leucocytes (72.85 +/- 14.95 closely after preparation and 93.74 +/- 5.91 24 hours after preparation) are consistent with data in the literature. The sterility of the solution and its influence on leucocyte vitality are also satisfactory. All work was done within the framework of preclinical tests of the mentioned preparation, as it was not yet approved for clinical use. In view of the assessed properties there are prerequisite conditions for its early use in clinical practice.