Effect of temperature on the development of chromosome aberrations in human blood lymphocytes irradiated in pulse and continuous modes at BARS-6 reactor

Summarized results of 5 repeated experiments of comparative study of radiation effects of the pulse reactor BARS-6 either in a single pulse or a continuos irradiation mode on human lymphocytes are presented. Higher efficiency (30-40%) of continuous irradiation (exposure duration 1 h) rather than pulse irradiation with ultra-high dose rates (1-2.5) x 10(6) Gy/min (pulse duration 65 micros) was confirmed. The efficiency ratio did not depend on the temperature, 20 degrees C or 0 degrees C, during the exposure. Cell repair system and chromatin conformation influence on the results obtained is discussed.     

Study of dose-rate effects of neutron radiation in a wild-type and a repair-deficient yeasts saccharomyces

We report here a comparative analysis of RBE for lethality of a single pulse (duration 65 micros) of fast neutron with ultra high dose rates (up to 6 x 10(6) Gy/s) and continuous neutron radiation (3.6 x 10(3) s) of the pulse reactor BARS-6. Three diploid strains, one haploid strain and three diploid repair-deficient strains (rad52-1/rad52-1; rad54/rad54; rad2/rad2) were used. The RBE values (D(0gamma)/1D(0n)) of a single pulse and continuous neutron irradiation were equal (1.7-1.8) with maximum RBE (4.1-3.1) in region of low doses (shoulder region). Haploid cells were found to be more (3 times) sensitive to both gamma-rays and neutrons than the wild type. There was no obvious decrease in the RBE of 1.9 in highly sensitive haploid cells as compared with highly resistant diploid cells. The repair-deficient strains (rad52-1/rad52-1; rad54/rad54) were more (up to 10 fold) sensitive to both neutrons and gamma-rays as compared with their parent line. The RBE values of 1.5-1.7 of neutrons for these mutants (independent by of the mode of irradiation) were found. The repair-deficient mutant rad2/rad2 had similar sensitivity as a wild type and a RBE value was 2.0. We have concluded that biological effectiveness of the neutrons of pulse reactor BARS-6 was independent of the dose-rate, differing up to 10(8) fold. The RBE didn't vary significantly with the capacity of cells to repair DNA damages.     

Life expectency and fertility of postembrio stage drosophila after the pulse-periodic X-ray irradiation

The effect of 13 Hz repetition rate X-ray pulses with 3 x 10(-6)-1.5 x 10(-4) Gr per pulse dose during 5 minute on drosophila's larvae and on pupae vas investigated. It was shown that the effect depends on drosophila's age as well as on X-ray dose and manifests itself in variation of life expectancy and fertility.     

Dose rate effect on micronuclei induction in human blood lymphocytes exposed to single pulse and multiple pulses of electrons

The effects of single pulses and multiple pulses of 7 MV electrons on micronuclei (MN) induction in cytokinesis-blocked human peripheral blood lymphocytes (PBLs) were investigated over a wide range of dose rates per pulse (instantaneous dose rate). PBLs were exposed to graded doses of 2, 3, 4, 6, and 8 Gy of single electron pulses of varying pulse widths at different dose rates per pulse, ranging from 1 × 10⁶ Gy s⁻¹ to 3.2 × 10⁸ Gy s⁻¹. Different dose rates per pulse were achieved by changing the dose per electron pulse by adjusting the beam current and pulse width. MN yields per unit absorbed dose after irradiation with single electron pulses were compared with those of multiple pulses of electrons. A significant decrease in the MN yield with increasing dose rates per pulse was observed, when dose was delivered by a single electron pulse. However, no reduction in the MN yield was observed when dose was delivered by multiple pulses of electrons. The decrease in the yield at high dose rates per pulse suggests possible radical recombination, which leads to decreased biological damage. Cellular response to the presence of very large numbers of chromosomal breaks may also alter the damage.     

The role of the DMPO-hydrated electron spin adduct in DMPO-*OH spin trapping

Time-resolved in situ radiolysis ESR (electron spin resonance, equivalently EPR, electron paramagnetic resonance) studies have shown that the scavenging of radiolytically produced hydroxyl radical in nitrous oxide-saturated aqueous solutions containing 2 mM DMPO is essentially quantitative (94% of the theoretical yield) at 100 micros after the electron pulse [1]. This result appeared to conflict with earlier results using continuous cobalt-60 gamma radiolysis and hydrogen peroxide photolysis, where factors of 35 and 33% were obtained, respectively [2,3]. To investigate this discrepancy, nitrogen-saturated aqueous solutions containing 15 mM DMPO were cobalt-60 gamma irradiated (dose rate = 223 Gy/min) for periods of 0.25-6 min, and ESR absorption spectra were observed approximately 30 s after irradiation. A rapid, pseudo-first-order termination reaction of the protonated DMPO-hydrated electron adduct (DMPO-H) with DMPO-OH was observed for the first time. The rate constant for the reaction of DMPO-H with DMPO-OH is 2.44 x 10(2) (+/- 2.2 x 10(1)) M(-1) s(-1). In low-dose radiolysis experiments, this reaction lowers the observed yield of DMPO-OH to 44% of the radiation-chemical OH radical yield (G = 2.8), in good agreement with the earlier results [2,3]. In the absence of the DMPO-H radical, the DMPO-OH exhibits second-order radical termination kinetics, 2k(T) = 22 (+/- 2) M(-1) s(-1) at initial DMPO-OH concentrations > or = 13 microM, with first-order termination kinetics observed at lower concentrations, in agreement with earlier literature reports [4].     

In vitro and in vivo expression of endothelial von Willebrand factor and leukocyte accumulation after fractionated irradiation

Previous investigations have demonstrated an increased release of von Willebrand factor (VWF; also known as vWF) in endothelial cells after high single-dose irradiation in vitro. We have also found increased levels of Vwf protein in mouse glomeruli after a high single dose of renal irradiation in vivo. In addition, increased numbers of leukocytes were observed in the renal cortex after irradiation in vivo. The aim of the present study was to investigate and quantify these biological processes after clinically relevant fractionated irradiation and to relate them to changes in renal function. A significantly greater increase in release of VWF was observed in cultured human umbilical vein endothelial cells (HUVECs) after fractionated irradiation (20 x 1.0 Gy) than after a single dose of 20 Gy (147% compared to 115% of control, respectively, P < 0.0005). In contrast with the in vitro observations, glomerular Vwf staining was lower after fractionated irradiation in vivo (20 x 2.0 Gy or 10 x 1.6 Gy +/- re-irradiation) than after a single dose of 16 Gy. The number of leukocytes accumulating in the renal cortex was also lower after fractionated in vivo irradiation than after a single radiation dose. The onset of these events preceded renal functional and histopathological changes by approximately 10 weeks. These data indicate that radiation-induced changes in endothelial VWF expression after in vivo irradiation may be distinct from the in vitro observations. Increased VWF expression may reflect pivotal processes in the pathogenesis of late radiation nephropathy and provide a clue to appropriate timing of pharmacological intervention.     

Influence of He-Ne laser irradiation and local anesthetics on potassium channels in the snail neurons

Using clamp method it had been shown that He-Ne laser irradiation of the snail neurons increases the amplitude of voltage-gated slow potassium currents in dose of 0.7 x 10(-4) (fluence 1.5 x 10(2) Wt/m2) and decreases it in dose 0.7 x 10(-3). Bupivacaine and lidocaine suppressed potassium currents. Laser irradiation in dose 0.7 x 10(-3) enhanced the inhibitory effect of bupivacaine (10 mcM) and in dose 0.7 x 10(-4) it decreased the inhibitory effect of bupivacaine. The results of the study suggest mechanisms of the He-Ne laser irradiation effect in combination with pharmacological substances on ion channels of electrically excitable cells.     

Concordant induction of rapid in vivo pulsatile insulin secretion by recurrent punctuated glucose infusions

Insulin is largely secreted as serial secretory bursts superimposed on basal release, insulin secretion is regulated through changes of pulse mass and frequency, and the insulin release pattern affects insulin action. Coordinate insulin release is preserved in the isolated perfused pancreas, suggesting intrapancreatic coordination. However, occurrence of glucose concentration oscillations may influence the process in vivo, as it does for ultradian oscillations. To determine if rapid pulsatile insulin release may be induced by minimal glucose infusions and to define the necessary glucose quantity, we studied six healthy individuals during brief repetitive glucose infusions of 6 and 2 mg x kg(-1) x min(-1) for 1 min every 10 min. The higher dose completely synchronized pulsatile insulin release at modest plasma glucose changes ( approximately 0.3 mM = approximately 5%), with large ( approximately 100%) amplitude insulin pulses at every single glucose induction (n = 54) at a lag time of 2 min (P < 0.05), compared with small (10%) and rare (n = 3) uninduced insulin excursions. The smaller glucose dose induced insulin pulses at lower significance levels and with considerable breakthrough insulin release. Periodicity shift from either 7- to 12-min or from 12- to 7-min intervals between consecutive glucose (6 mg x kg(-1) x min(-1)) infusions in six volunteers revealed rapid frequency changes. The orderliness of insulin release as estimated by approximate entropy (1.459 +/- 0.009 vs. 1.549 +/- 0.027, P = 0.016) was significantly improved by glucose pulse induction (n = 6; 6 mg x kg(-1) x min(-1)) compared with unstimulated insulin profiles (n = 7). We conclude that rapid in vivo oscillations in glucose may be an important regulator of pulsatile insulin secretion in humans and that the use of an intermittent pulsed glucose induction to evoke defined and recurrent insulin secretory signals may be a useful tool to unveil more subtle defects in beta-cell glucose sensitivity.     

Pulsed-dose-rate and low-dose-rate brachytherapy: comparison of sparing effects in cells of a radiosensitive and a radioresistant cell line

Pulsed-dose-rate regimens are an attractive alternative to continuous low-dose-rate brachytherapy. However, apart from data obtained from modeling, only a few in vitro results are available for comparing the biological effectiveness of both modalities. Cells of two human cell lines with survival fractions of 80% (RT112) and 10% (HX142) after a single dose of 2 Gy and with different halftimes for split-dose recovery and low-dose recovery were used. The cells were irradiated with a continuous low dose rate (80 cGy per hour) or with pulsed dose rate. Two different pulsed dose rates were tested: 4.25 Gy/h and 63 Gy/h. The effects of dose per pulse and the length of the interval between the pulses were investigated while keeping the overall treatment time constant. Survival after low-dose-rate irradiation was indistinguishable from that after pulses of 4.25 Gy/h in cells of both cell lines. Survival decreased with increasing dose per pulse. When the dose rate during the pulses was increased, survival decreased even further. This effect was most pronounced for the radiosensitive HX142 cells. In clinical pulsed-dose-rate brachytherapy, iridium sources move stepwise through the implant and deliver pulses at a high dose rate locally. These high-dose-rate pulses produce greater biological effectiveness compared to continuous low dose rate; this should be taken into account.     

Fast repair of deoxynucleotide radical cations by phenylpropanoid glycosides (PPGs) and their analogs

The repair effects on deoxynucleotide radical cations of phenylpropanoid glycosides (PPGs) and their analogs, isolated from a Chinese folk medicinal herb, were studied using the pulse radiolysis technique. The radical cations of deoxynucleotides were formed by the reaction of SO4*- with deoxynucleotides. On pulse irradiation of a nitrogen saturated deoxynucleotide aqueous solution containing 20 mM K2S2O8, 200 mM t-BuOH and one of the PPGs or their analogs, the transient absorption spectra of the radical cations of nucleotide decayed with the formation of those of the radical cation of PPGs or their analogs within several tens of microseconds after electron pulse irradiation. The result indicates that deoxynucleotide radical cations can be repaired by PPGs or their analogs. The rate constants of the repair reactions were determined to be 0.48-1.1 x 10(9), 0.64-1.80 x 10(9) and 2.12-4.4 x 10(9) M(-1) s(-1) for dAMP, dGMP and dCMP radical cations respectively. It is obvious that the rate constants of the repair reaction depend on the number of phenolic hydroxyl groups contained in the PPGs and their analogs. A deeper understanding of this new repair mechanism will undoubtedly help researchers design strategies to prevent and/or intervene more effective in free radical related diseases.     

Fast repair of dAMP radical anions by phenylpropanoid glycosides and their analogs

Repair effect on 2'-deoxyadenosine-5'-monophosphate (dAMP) radical anions by phenylpropanoid glycosides (PPGs) and their analogs, isolated from Chinese folk medicinal herb, was studied using pulse radiolysis technique. The radical anion of dAMP was formed by the reaction of hydrated electron with dAMP. On pulse irradiation of nitrogen-saturated dAMP aqueous solution containing 0.2 M t-BuOH and one of PPGs or their analogs, the transient absorption spectrum of the radical anion of dAMP decayed with the formation of that of the radical anion of PPGs or their analogs within several decades of microseconds after electron pulse irradiation. The results indicated that dAMP radical anions can be repaired by PPGs or their analogs. The rate constants of the repair reactions were deduced to be 1.6-4.5 x 10(8) M(-1) s(-1).     

Peculiarities of the influence of chemical and physical factors on cytogenetic indices of root meristem in Pisum sativum L

The results of comparative analysis of cytogenetic effects of various chemical and physical factors on meristemic cells of field pea (Pisum sativum L.) are presented. Cytogenetic effects of aluminum, cadmium and selenium compounds as well as X-ray irradiation were studied. Cytogenetic indices were investigated for different doses of the above factors and peculiarities of their influence on the ratio of mitotic phases were analyzed. The increasing in the proportion of cells at the specific mitotic stage was revealed. There were anaphase stage for aluminum chloride, prophase stage for cadmium chloride, meta- and anaphase stages for sodium selenate, and telophase stage for X-rays. Investigated chemical elements may be ranged according to their ability to induce the aberrant anaphase frequency in the row: Se (from 3.75 x 10(-6) M) > A1 (from 3.86 x 10(-5) M) > Cd (from 8.44 x 10(-5) M). Maximal experimental dose of X-ray irradiation (9.03 x 10(-3) C/kg) induces the frequency of chromosome aberration similar to those induced by maximal experimental doses of aluminum and selenium (3.86 x 10(-4) M and 8.34 x 10(-6) M).     

Fertilizing ability in vitro of golden hamster spermatozoa after acute testicular X-irradiation]

The present study is concerned with the effect of radiation to the testis on fertilizing ability in vitro using golden hamster spermatozoa. Male hamsters at 6 and 8 weeks of age were given acute testicular X-irradiation (200 kVp, 20 mA, 0.47-0.48 Gy/min). Spermatozoa were collected from the cauda epididymides at different times after irradiation and then they were suspended in fertilization medium. After preincubation for 4-5 hr, the spermatozoa were cultured with the eggs collected from mature hamsters treated with PMSG-hCG. Fertilized eggs were examined for incidence of sperm penetration and formation of pronuclei at 4-5 hr after insemination. The fertilization rate (47.7%) at the 6th week after irradiation with a dose of 2 Gy was much lower in comparison with the control value (92.6%). However, the fertilization rates at the 3rd and 9th weeks after irradiation were 97.7 and 90.6%, respectively. In these period, no difference was found between the irradiated groups and the control groups. From the changes in sperm concentration after irradiation with a dose of 2 Gy, it was found that the fertilization rate was the lowest at the 6th week. The sensitive stage to radiation during spermatogenesis with reference to the reduction of fertilizing ability after irradiation coincides with that of decrease in the sperm concentration and sperm motility. The results of fertilization rate at the 6th week after different doses of X-irradiation (0.25-6 Gy) indicated that the reduction of fertilization rate is nearly expressed as a dose-response relationship.(ABSTRACT TRUNCATED AT 250 WORDS)     

Radioprotective action of gas hypoxia (8%) in local irradiation of the kidneys]

The radioprotective action of hypoxia in local irradiation of kidneys was studied. The experiments were carried out on hybrid mice (CBA x C57Bl) exposed to local X-ray irradiation through a lead diaphragm matched to the kidneys. In the experimental group the animals breathed in gaseous mixture with 8% of oxygen prior to and during irradiation. 20-36 weeks after irradiation the functions of glomerular and juxtaglomerular systems, as well as mass of native and dried kidneys were studied in the animals. It is shown that hypoxia efficiently protects kidneys against irradiation and permits increasing the supplied dose of radiation at least by 25%. High-efficient protection is retained when passing from single to fractionated irradiation.     

Dosimetry of total skin electron irradiation

Elaboration of such a simple technique for total skin electron irradiation which ensures good dose homogeneity and minimal x-ray background dose. MATERIALS AND METHODS: We started large electron field irradiations with the Neptun 10p linear accelerator in the National Institute of Oncology -Budapest in 1986. After the installation of the Siemens Mevatron KD linear accelerator it was possible to introduce the modified Stanford technique. This technique satisfies better the requirements given in the objective. The required field size of 200x75 cm is produced as a result of two fields with 30 degrees angular separation (dual field) at a source skin distance of 465 cm. The patient's body is exposed to six dual electron fields. The electron energy is 6 MeV. Despite the long source skin distance the treatment time is relatively short due to the high dose rate (940 mu/min) capability of our Mevatron KD. The in air dose profiles were measured in miniphantom with semiconductor detector. Depth dose curves were measured in water and in polystyrene phantom with semiconductor detector and with films. RESULTS: The measured dose homogeneity of the 6 MeV energy dual field with 30 degrees angular separation is within +/- 5%in a 200x75cm plane field. The depth of dose maximum of the resulting dose distribution of six dual field irradiation is between 2 mm and 5 mm, while the depth of 80% isodose curve is about 8 mm. The total body x-ray background dose is less than 1% of the skin dose. CONCLUSION: The modified Stanford technique adapted to our Mevatron KD linear accelerator is suitable for total skin electron beam therapy.     

Regenerative effects of tetrachlorodecaoxide in BD IX rats after total-body gamma irradiation

Tetrachlorodecaoxide (TCDO) was tested for its effects in BD IX rats when combined with a single dose nearing LD50 of total-body irradiation (gamma rays, 60Co). In pilot tests we found that TCDO administrations prior to or immediately after irradiation led to a very high mortality rate (up to 90%), whereas the initiation of TCDO treatment on Day 2, 3, or 4 after irradiation lowered the death rate noticeably, with optimum results when TCDO application was started on Day 4. In our major experiment on 100 BD IX rats, it was demonstrated that the following treatment schedule considerably decreased the death rate (from 44 to 4%): 15.5 mumol TCDO/kg body wt/day on Days 4-6 after irradiation and 7.75 mumol/kg body wt/day on Days 7-11. The animals treated with TCDO showed only mild anemia in the peripheral blood, accompanied by reticulocytosis and low-grade leukocytopenia. Examination of the bone marrow on Day 12 after irradiation revealed X-ray-induced agranulocytosis in the animals that had received only physiological saline solution, whereas in the bone marrow of the animals treated with TCDO there was erythropoiesis as well as myelopoiesis. In addition, the degree of hair loss and depigmentation occurring about 1 month after irradiation was considerably reduced by TCDO. From these results it can be postulated that TCDO has two different effects: as an oxygen donator it causes radiosensitization in the tissue when given before or immediately after irradiation; as an agent stimulating phagocytes and tissue regeneration, it promotes regeneration very effectively when damage is already evident in the tissue.     

Dose-response relationship for translocation induction in spermatogonia of the crab-eating monkey (Macaca fascicularis) by chronic gamma-ray-irradiation

The induction of reciprocal translocations in spermatogonia of the crab-eating monkey (Macaca fascicularis) by chronic gamma-irradiation (1.8 x 10(-5) Gy/min, about 0.024 Gy/22 h/day) was examined. The frequencies of translocation per cell were 0.15% at 0.3 Gy, 0.27% at 1.0 Gy and 0.33% at 1.5 Gy. The dose-response relationship for translocation yield was a linear one with a regression coefficient (b) of 0.16 x 10(-2). When the slope (b) of the regression line was compared with that at a high dose rate (0.25 Gy/min, b = 1.79 x 10(-2), it was clear that the induction rate of translocations after chronic gamma-irradiation was only about one-tenth of that after high-dose-rate irradiation. Thus, there was evidence for a pronounced dose-rate effect in the crab-eating monkey.     

The radioprotective effect of extracts of Archangelica officinalis Hoffm. and Ledum palustre L. on mice

A single injection of Archangelica officinalis Hoffm. and Ledum palustre L. extracts to mice 5-15 min before irradiation with a median lethal dose increased their survival rate. The most favourable effect was produced by a combination of the two preparations: by day 30 100% of animals survived after a dose of 6 Gy (LD50/30); 70% survived after a dose of 7.5 Gy (LD90/30), and 25% after a dose of 8 Gy (LD100/12). DMF for the extract mixture was 1.48.     

Impact of pulsed Nd:YAG laser on the marine biofilm-forming bacteria Pseudoalteromonas carrageenovora: significance of physiological status

The impact of pulsed Nd:YAG (neodymium-doped yttrium/aluminium garnet) laser irradiation on the marine biofilm-forming bacteria Pseudoalteromonas carrageenovora during two growth stages (log phase and stationary phase) and under two stresses (reduced temperature and nutrient limitation) was investigated. Bacteria were exposed to a laser fluence of 0.1 J x cm(-2) for 5, 10, and 15 min with a peak power of 20 MW x cm(-2), a pulse width of 5 ns, and an average power of 1 W x cm(-2) with a repetition rate of 10 Hz. The mortality of bacteria immediately after the irradiation as well as after a set period of time was determined. Mortality was higher among log-phase bacteria (72%) than bacteria in the stationary phase (51%) and those grown under nutrient limitation (51%). Bacteria grown at reduced temperature had a mortality of 49%. However, the differences in cell density of log-phase, stationary-phase, nutrient-limited, and low-temperature irradiated samples compared with controls after 5 h of incubation were 96, 93, 94, and 86%, respectively. The mortality values suggest that the same laser fluence has different degrees of effectiveness, depending on the physiological state of the bacteria.     

Low semen dose intracornual insemination of cows at fixed time after PGF2alpha treatment or at spontaneous estrus

The numbers of spermatozoa per insemination and the site of semen deposition in the uterine horn appear to interact to influence pregnancy rate. In two experiments, the effect of a single low dose (2 x 10(6) spermatozoa) intracornual insemination (LD-ICI) on bovine pregnancy rate was compared with that of intracornual (SD-ICI) and conventional (SD-AI) inseminations of 40 x 10(6) spermatozoa. In Experiment 1, 157 cows were treated twice with PGF(2)alpha at a 14-day interval and inseminated at a fixed time (80-82 h) after the second PGF(2)alpha injection using LD-ICI (n=44), SD-ICI (n=61) or SD-AI (n=52). In LD-ICI and SD-ICI groups, semen was deposited in the horn ipsilateral to the ovulatory follicle close to the utero-tubal junction (LD-ICI-UTJ, n=33 and SD-ICI-UTJ, n=41) or in the middle part of the horn (LD-ICI-MH, n=11 and SD-ICI-MH, n=20). Pregnancy rates after LD-ICI-UTJ, LD-ICI-MH, SD-ICI-UTJ and SD-ICI-MH were 27%, 27%, 39% and 35%, respectively (P>0.05). The total pregnancy rate after LD-ICI (27%) did not differ (P>0.05) from that after SD-ICI (37%) or SD-AI (34%). In Experiment 2 (field trial), 362 cows were allotted, at spontaneous estrus, to LD-ICI-UTJ (n=86), LD-ICI-MH (n=97) or SD-AI (n=179). Pregnancy rates after LD-ICI and SD-AI were 47% and 45%, respectively (P>0.05). After LD-ICI-UTJ, the pregnancy rate (54%) did not differ significantly (P>0.05) to that obtained after LD-ICI-MH (41%) and after SD-AI (45%). The results of the study show that the single intracornual insemination of cows with 2 x 10(6) spermatozoa at fixed time, 80-82 h after the second PGF(2)alpha injection or at spontaneous estrus resulted in similar pregnancy percentage as intracornual and conventional inseminations with 40 x 10(6) spermatozoa per semen dose. With intracornual insemination using low or standard dose of spermatozoa, the pregnancy rates were not significantly affected by the exact site of semen deposition in the uterine horn, near the utero-tubal junction or in the middle part.