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.

Incidence of cutaneous malignant melanoma in the Czech Republic: the risks of sun exposure for adolescents.

The Czech Republic reported one of the highest incidence rate in cutaneous melanoma (CM) in Europe and because this incidence has been increasing, mainly among young people, the main goal of our study was to establish sun exposure behavior risk factors for CM formation and to evaluate whether the young generation of Czechs is exposed to a higher risk of CM than the older generation. A questionnaire-based case-control study was conducted. We obtained 978 completed questionnaires: 216 from patients with CM and 762 from healthy respondents. The healthy individuals were further divided to adolescents (n = 460) and older respondents (n = 302). Three logistic regression models were developed: 1. patients with CM vs. healthy older respondents, 2. adolescents vs. healthy older respondents, and 3. patients with CM vs. adolescents. The main risk factors for all three models were the number of sunburn episodes and the use of the sunscreen in the childhood. The most alarming results for adolescents included: all day sun exposure, including times of maximum risk (11 AM to 3 PM), inadequate use of sunscreen in adulthood, and frequent mountain holidays. Our results show that sun-safety in the young generation is satisfactory, when the responsibility for sun exposure behavior is in the hands of their parents; however, when children become adolescents, they become immune to sun-safety and risk prevention campaigns and their behavior becomes much more risky. Our results further suggest the sun-safety campaigns need to be modified in such a way as to have greater impact and influence on adolescent sun-risk behaviors.

Wirelessly Powered Endoscopically Implantable Devices into the Submucosa as the Possible Treatment of Gastroesophageal Reflux Disease.

Gastroesophageal reflux disease (GERD) is a rather common disease with a prevalence reaching up to 10 or 20% in the western world. The most specific symptoms which point to the diagnosis of GERD are feelings of heartburn and the regurgitation of acidic stomach contents into the esophagus. However, a certain number of patients do not respond to standard therapy, and in these cases, it is necessary to resort to other treatment methods, such as laparoscopic fundoplication or electrostimulation of the lower esophageal sphincter. The aim of our work was to design and manufacture a miniature, battery-less stimulator to provide electric stimulation of the lower esophageal sphincter, which could be implanted deep into the submucosa of the distal esophagus. The main goal was to provide a battery-less system as opposed to traditional battery neurostimulators to reduce the size and weight of the device. An electronic prototype of a wirelessly powered implantable device was developed. We used animal models for the experiments. The device is designed to treat GERD via electrical stimulation of the muscularis propria. It is implanted into the submucosal pocket by the lower esophageal sphincter with an endoscope. This method of implantation is superior to esophageal stimulators used today because of very low invasiveness of the surgery. Bipolar neurostimulation via two gold-plated leads is provided. The device does not have any source of energy; it is powered wirelessly which reduces the risk of potential battery leakage and reduces the overall dimensions.

Immunological response in the mouse melanoma model after local hyperthermia.

Our study was aimed to characterize the phenotype and functional endpoints of local microwave hyperthermia (LHT, 42 degrees C) on tumor infiltrating and spleen leukocytes. The effectiveness of LHT applied into the tumor of B16F10 melanoma-bearing C57/BL6 mice was compared with anesthetized and non-treated animals. Subpopulations of leukocytes were analyzed using the flow cytometry, and the cytotoxic activity of splenocytes against syngeneic B16F10 melanoma and NK-sensitive YAC-1 tumor cell lines was evaluated in (51)Cr-release assay. Similarly, the in vitro modification of the heat treatment was performed using healthy and melanoma-bearing splenocytes. We found a 40 % increase of activated monocytes (CD11b+CD69+) infiltration into the tumor microenvironment. In the spleen of experimental animals, the numbers of cytotoxic T lymphocytes (CTLs-CD3+CD8+) and NK cell (CD49b+NK1.1+) raised by 22 % and 14 %, respectively, while the NK1.1+ monocytes decreases by 37 %. This was accompanied by an enhancement of cytotoxic effector function against B16F10 and YAC-1 targets in both in vivo and in vitro conditions. These results demonstrate that LHT induces better killing of syngeneic melanoma targets. Furthermore, LHT evokes the homing of activated monocytes into the tumor microenvironment and increases the counts of NK cells and CTL in the spleen.

Temperature dependence of blood surface tension.

Whole blood surface tension of 15 healthy subjects recorded by the ring method was investigated in the temperature range from 20 to 40 degrees C. The surface tension omega as a function of temperature t ( degrees C) is described by an equation of linear regression as omega(t) = (-0.473 t + 70.105) x 10(-3) N/m. Blood serum surface tension in the range from 20 to 40 degrees C is described by linear regression equation omega(t) = (-0.368 t + 66.072) x 10(-3) N/m and linear regression function of blood sediment surface tension is omega(t) = (-0.423 t + 67.223) x10(-3) N/m.

Polycaprolactone foam functionalized with chitosan microparticles - a suitable scaffold for cartilage regeneration.

For biodegradable porous scaffolds to have a potential application in cartilage regeneration, they should enable cell growth and differentiation and should have adequate mechanical properties. In this study, our aim was to prepare biocompatible scaffolds with improved biomechanical properties. To this end, we have developed foam scaffolds from poly-epsilon-caprolactone (PCL) with incorporated chitosan microparticles. The scaffolds were prepared by a salt leaching technique from either 10 or 15 wt% PCL solutions containing 0, 10 and 20 wt% chitosan microparticles, where the same amount and size of NaCl was used as a porogen in all the cases. PCL scaffolds without and with low amounts of chitosan (0 and 10 wt% chitosan) showed higher DNA content than scaffolds with high amounts of chitosan during a 22-day experiment. 10 wt% PCL with 10 and 20 wt% chitosan showed significantly increased viscoelastic properties compared to 15 wt% PCL scaffolds with 0 and 10 wt% chitosan. Thus, 10 wt% PCL scaffolds with 0 wt% and 10 wt% chitosan are potential scaffolds for cartilage regeneration.

Surface tension of blood.

The surface tension of blood assessed in a group of 71 healthy subjects (24 men and 47 women) by the drop method at a temperature of 22 degrees C was 55.89 x 10(-3) N x m(-1), S.D.=3.57 x 10(-3) N x m(-1). It did not correlate with age or sex of the examined subjects nor with any of the following variables: red cell sedimentation rate, blood haemoglobin levels, number of erythrocytes, total serum cholesterol, total serum triacylglycerols, creatinine blood levels, ALT and AST activity. The surface tension of blood and other body fluids can play an important part not only in the genesis and development of decompression sickness but also in other processes in the organism.

Influence of dynorphin A (1-13) and dynorphin A (1-10) amide on stress-induced analgesia.

Dynorphin A (1-13) and its analog dynorphin A (1-10) amide were applied intracerebroventricularly in male ICR mice. Both dynorphins did not reveal any analgesic activity in tail-flick test under normal (non-stressed) conditions. However, in combination with stress (forced swimming or whole body vibration) both dynorphins prolonged tail-flick latencies when compared with stressed saline controls. Naloxone inhibited the effect of dynorphins in forced swimming test. Neither dynorphin A (1-13) nor dynorphin A (1-10) amide increased tail-flick latencies when combined with weak immobilization stress. Our results suggest that the analgesic effects of dynorphins are potentiated by strong stressors.

Effect of agroclavine on NK activity in vivo under normal and stress conditions in rats.

Agroclavine is a natural, clavine type of ergot alkaloid with D1 dopamine and a-adrenoceptor agonistic properties. We showed previously that in vitro agroclavine enhances natural killer (NK) cell activity, increases interleukin-2 and interferon-gamma production and prolongs the survival time of tumor-bearing mice. The aim of this study was 1) to test the effect of agroclavine on NK activity in vivo, and 2) to assess the potential toxicity of high doses of agroclavine on cardiac and liver functions using creatine kinase MB (CKMB) and alanine aminotransferase (ALT) as biochemical markers in normal and stressed animals. The effect of stress was studied because we examined promising anticancer properties of agroclavine and malignant diseases are supposed to be a potent stressful event for patients. In our experiments 3-month-old male rats of the Wistar-Kyoto strain were used. Agroclavine was injected intraperitoneally (0.5 mg/kg or 0.05 mg/kg) 30 min before stress (four hours' restraint and immersion in 23 degrees C water). The animals were killed 30 min after stress, blood was collected and the spleen was removed. Non-stressed animals treated with agroclavine were killed 5 h after the drug administration. The results confirmed our previous in vitro results and showed that also in vivo agroclavine increases NK cell activity under non-stress conditions. Agroclavine only slightly increased CKMB and had no influence on ALT in non-stressed animals. These promising results are limited by the fact that agroclavine (0.5 mg/kg) diminished NK cell activity and significantly increased ALT and CKMB under stress conditions.

The effect of drugs on the mortality of mice inoculated with Friend leukaemia virus or toxoplasma gondii.

Infection and tumors provoke substantial changes accompanied with the disbalance of many neuroendocrine factors which in their summarizing effects influence the life span of animals. Our previous results showed enhanced mortality after one injection of morphine in association with Friend leukaemia virus infection. The aim of this study was to examine the effects of some other opioids (pethidine and pentazocine) and an acetylcholine esterase inhibitor neostigmine on the survival of animals under two conditions: (1) Friend leukaemia virus infection which mostly depressed immune functions, and (2) Toxoplasma gondii infection which in general enhanced the immune status. In contrast to our previous observation with morphine, the mortality induced by single doses of pethidine (150 mg/kg) or pentazocine (50-75 mg/kg) was unchanged during the Friend leukaemia virus infection. A single injection of neostigmine (0.42 or 0.56 mg/kg) was significantly more lethal in DBA-2 mice infected with Friend leukaemia virus. Neostigmine in doses of 0.33 and 0.4 mg/kg caused death in 46 % and 57 %, respectively, of animals infected with Toxoplasma gondii which was significantly higher in comparison with only 8 % and 12.5 % in control groups. Pethidine (150 mg/kg) killed 70 % of Toxoplasma gondii infected animals and even 90 % of non-infected mice. Thus, the Friend leukaemia virus and Toxoplasma gondii infections increased toxicity only of some drugs which may, at least partly, be associated with altered immune status during infection and involvement of the cholinergic system.

Exposure to stress alters the effects of dynorphins in the hot plate test.

The analgesic effects of intracerebroventricularly (i.c.v.) administered dynorphin A(1-13) and its analog dynorphin A(1-10)amide using the hot plate test were studied in mice. Both dynorphins applied i.c.v. by the freehand method had an analgesic effect but no effect was seen when applied i.c.v. through an implanted cannula. Moreover, freehand i.c.v. injection of saline increased the time of immobility in the forced swimming test and glycaemia levels compared with intact mice. In contrast to the freehand injection, saline administration through an implanted cannula did not influence the immobility of animals in forced swimming test when compared with the intact controls. These results suggest that 1) the freehand method is very stressful procedure of administration which could influence the effects of dynorphins in the hot plate test and 2) dynorphins exert an analgesic effect in the hot plate test only when combined with a stressor (freehand i.c.v. injection).

Evaluación de la función muscular del cuádriceps en pacientes con EPOC modera-severa / Evaluation of quadricep muscle function in patients with moderate-severe COPD

Objetivo: el objetivo del estudio fue evaluar la función muscular periférica de pacientes con EPOC moderadasevera, mediante la medición de la fuerza muscular isométrica e isotónica del cuádriceps y la masa muscular mediante ecografía. Método: fueron incluidos pacientes con EPOC moderada-severa estables, remitidos a Rehabilitación Respiratoria. En la extremidad dominante, se midió la fuerza isométrica (dinamómetro hidráulico), test 1RM y se realizó una ecografía para obtener el área transversal y dimensiones de diversas secciones musculares. Resultados: participaron 34 pacientes con EPOC estable: GOLD 2 (n = 11), GOLD 3 (n = 12) y GOLD 4 (n = 11) Los GOLD 4 tenían un IMC P50 (P25 - P75) 25,3 (22,5 - 28,8), significativamente menor que los GOLD 3: 32,0 (27,5 - 34,0); p = 0,025. Se evidenció debilidad del cuádriceps (1RM/peso <0,60 1RM/IMC <120%), de forma global: 0,33 (0,22 - 0,41) y 93% (61 - 112) respectivamente, y en cada grupo, sin diferencias significativas entre ellos. Tampoco hubo diferencias significativas entre los grupos en las mediciones de ecografía muscular. La fuerza isotónica (1RM) se correlacionó con el grosor del cuádriceps QT (r = 0,529; p = 0,003) y el grosor del músculo vasto intermedio Q1(r = 0,514; p = 0,004) La fuerza isométrica se correlacionó con el QT (0,621; p <0,001), Q1 (0,441; p = 0,009) y el área transversal AQ (r = 0,587; p <0,001) Para cada grupo, el QT y la fuerza isométrica se correlacionaron significativamente. Conclusión: en nuestra serie de pacientes se demuestra la existencia de debilidad muscular del cuádriceps, independientemente del grado de severidad de la enfermedad y en estrecha relación con la masa muscular de dicho músculo. Palabras clave: disfunción muscular, fuerza isotónica, 1RM, fuerza isométrica, ecografía muscular

Objective: The objective of this study was to evaluate peripheral muscle function in patients with moderate-severe COPD by measuring isometric and isotonic muscular strength of the quadriceps and muscle mass through ultrasound. Method: This study included stable patients with moderate-severe COPD who were referred to pulmonary rehabilitation. In the dominant extremity, isometric strength (hydraulic dynamometer) and the 1RM test were measured and an ultrasound was done to determine the transversal area and dimensions of different sections of muscle. Results: 34 stable patients with COPD participated: GOLD 2 (n = 11), GOLD 3 (n = 12) and GOLD 4 (n = 11). GOLD 4 patients had a BMI P50 (P25 - P75) 25.3 (22.5 - 28.8), which was significantly lower than GOLD 3 patients: 32.0 (27.5 - 34.0); p = 0.025. Weakness in the quadriceps was observed (1 RM/weight <0.60; 1 RM/BMI <120%) overall: 0.33 (0.22 - 0.41) and 93% (61 - 112), respectively, and in each group, without significant differences between groups. There were also no significant differences between groups for ultrasound muscle measurements. Isotonic strength (1 RM) correlated to quadricep thickness QT (r = 0.529; p = 0.003) and vastus intermedius muscle thickness Q1(r = 0.514; p = 0.004). Isometric strength correlated to QT (0.621; p <0.001), Q1 (0.441; p = 0.009) and transversal area AQ (r = 0.587; p <0.001). QT and isometric strength were significantly correlated in each group. Conclusion: In our study, patients showed muscular weakness in the quadriceps, regardless of the degree of disease severity and this weakness was closely related to the mass of said muscle

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.

Biliary excretion of 110mAg and its kinetics in the isolated perfused liver in rats.

The biliary excretion of 110mAg in rats after i.v. administration of an aqueous solution of 110mAgNO3 (4.57 micrograms; 16kBq per rat) was studied for a period of 24 hours. The maximum rate of excretion was reached in 30th minute after the metal administration and over 70% of the silver dosed was excreted during 24 hours. Using the method of isolated perfused liver it was observed that 110mAg is rapidly taken up in the liver. During the five minutes period of the perfusion less than 50% of silver administered was found in the perfusion medium. In following minutes the level of the metal in the medium remained approximately constant. It was suggested that the rate of excretion of silver and its high uptake in the liver tissue is in connection with an unusual binding of it in the bile.

Reinfusion of UV-irradiated blood in the treatment of infection. I. Experimental infection with staphylococci.

During the last 10 years, reinfusion of UV-irradiated blood has been rediscovered again as a therapeutic method suitable in the treatment of a variety of diseases. The described series of model experiments on rabbits confirm its beneficial effects in the treatment of staphylococcal infection: the control animals reinfused blood not exposed to UV radiation died all within 48 hours after injected with a suspension of live Staphylococcus aureus culture; all rabbits reinfused UV-irradiated blood (2 ml per kg body weight) survived the whole period of observation (30 days); reinfusion of UV-irradiated blood in a volume reduced to 1 ml.kg-1 body weight prolonged the animals' life-span to 96 or 120 hours.

The effect of 110mAg on ceruloplasmin oxidase activity in rats.

The effect of single i.v. injection of 110mAgNO3 (0.183 mg Ag+ X kg-1 b.wt.) in rats on the ceruloplasmin oxidase activity (Cp) and copper serum concentration was studied. It was found that Cp activity in the serum decreased to 70% of the control value and simultaneously serum copper concentration decrease to 30% of the control level. In both cases the decrease was independent on the time elapsed after silver administration. Comparing these results with those reported recently in mice Cu deficit in the rat serum was approximately twice higher. This fact is considered to be an inter-species difference. The concentration of copper in the hepatic supernatant significantly decreased (to eight times from control value) after silver injection. Only less than 10% of the total amount of Ag found in whole liver was taken up to hepatic supernatant. GPC analysis of the supernatant (Sephadex G-75) revealed that no Ag-metallothionein fraction is present. On the basis of the results obtained it was concluded that the mechanism of silver inhibition of Cp oxidase activity remains still in question.

The effect of CdCl2 and AgNO3 pretreatment on the distribution of 110mAg+ in rat liver.

The distribution of 110mAg in the elution fractions obtained from gel filtration (Sephadex G-75) of the supernatants of rat liver homogenates was studied. 110mAgNO3 was administered i.v. at the doses 0.183 and 0.912 mg Ag+ . kg-1 . b . wt.and rats were killed 30 min after the administration. Differential centrifugation of the homogenates was performed to determine 110mAg+ distribution in subcellular structures of hepatocytes. The effect of CdCl2 and AgNO3 pretreatment (2 s.c. injections of 2.5 mg . kg-1 during 48 h) was investigated and results were compared with those obtained without pretreatment. It was found that AgNO3 pretreatment does not affect 110mAg+ distribution in the elution fractions and the major part of the metal is concentrated in the high-molecular-weight protein fraction. On contrary, after CdCl2 pretreatment almost all 110mAg+ is bound in the metallothionein (MT) fraction. Differential centrifugation revealed the main portion of the metal in nuclei and cell membranes and only small amount in lysosomal supernatant. After CdCl2 pretreatment the content of 110mAg+ in supernatant considerably increases. Results confirm the affinity of silver to MT and moreover show that this metal probably does not significantly induce the MT synthesis in the liver tissue.