Postexposure Week Research Articles

Transcriptomes of Wet Skin Biopsies Predict Outcomes after Ionizing Radiation Exposure with Potential Dosimetric Applications in a Mouse Model.

Countermeasures for radiation diagnosis, prognosis, and treatment are trailing behind the proliferation of nuclear energy and weaponry. Radiation injury mechanisms at the systems biology level are not fully understood. Here, mice skin biopsies at h2, d4, d7, d21, and d28 after exposure to 1, 3, 6, or 20 Gy whole-body ionizing radiation were evaluated for the potential application of transcriptional alterations in radiation diagnosis and prognosis. Exposure to 20 Gy was lethal by d7, while mice who received 1, 3, or 6 Gy survived the 28-day time course. A Sammon plot separated samples based on survival and time points (TPs) within lethal (20 Gy) and sublethal doses. The differences in the numbers, regulation mode, and fold change of significantly differentially transcribed genes (SDTGs, p < 0.05 and FC > 2) were identified between lethal and sublethal doses, and down and upregulation dominated transcriptomes during the first post-exposure week, respectively. The numbers of SDTGs and the percentages of upregulated ones revealed stationary downregulation post-lethal dose in contrast to responses to sublethal doses which were dynamic and largely upregulated. Longitudinal up/downregulated SDTGs ratios suggested delayed and extended responses with increasing IR doses in the sublethal range and lethal-like responses in late TPs. This was supported by the distributions of common and unique genes across TPs within each dose. Several genes with potential dosimetric marker applications were identified. Immune, fibrosis, detoxification, hematological, neurological, gastric, cell survival, migration, and proliferation radiation response pathways were identified, with the majority predicted to be activated after sublethal and inactivated after lethal exposures, particularly during the first post-exposure week.

Transcriptomics of Wet Skin Biopsies Predict Early Radiation-Induced Hematological Damage in a Mouse Model.

The lack of an easy and fast radiation-exposure testing method with a dosimetric ability complicates triage and treatment in response to a nuclear detonation, radioactive material release, or clandestine exposure. The potential of transcriptomics in radiation diagnosis and prognosis were assessed here using wet skin (blood/skin) biopsies obtained at hour 2 and days 4, 7, 21, and 28 from a mouse radiation model. Analysis of significantly differentially transcribed genes (SDTG; p ≤ 0.05 and FC ≥ 2) during the first post-exposure week identified the glycoprotein 6 (GP-VI) signaling, the dendritic cell maturation, and the intrinsic prothrombin activation pathways as the top modulated pathways with stable inactivation after lethal exposures (20 Gy) and intermittent activation after sublethal (1, 3, 6 Gy) exposure time points (TPs). Interestingly, these pathways were inactivated in the late TPs after sublethal exposure in concordance with a delayed deleterious effect. Modulated transcription of a variety of collagen types, laminin, and peptidase genes underlay the modulated functions of these hematologically important pathways. Several other SDTGs related to platelet and leukocyte development and functions were identified. These results outlined genetic determinants that were crucial to clinically documented radiation-induced hematological and skin damage with potential countermeasure applications.

Prunus armeniaca L (apricot) protects rat testes from detrimental effects of low-dose x-rays

Exposure to low x-ray doses damages the spermatozoa, mainly by late-onset (ie, after 3 months) oxidative stress. Antioxidants ameliorate oxidation and prevent tissue damage. Prunus armeniaca L (apricot), rich in carotenoids and vitamins, is a potent natural antioxidant. We hypothesized that an apricot-rich diet might ameliorate the detrimental effects of low-dose x-rays on testis tissue. A 20% apricot diet was composed isoenergetically to the regular rodent diet. The total phenolic content, reducing power, and antioxidant capacity of both diets were determined. Sprague-Dawley rats received apricot-rich diets before and after x-ray exposure. Regular diets were given to controls. Rats were exposed to 0.2 Gy x-rays at the eighth week and were euthanized at the 20th postexposure week. Testicular oxidative status was determined by tissue thiobarbituric acid-reactive substances, reduced glutathione, superoxide dismutase, and catalase activities. For histologic evaluation, qualitative and quantitative microscopic determinations were performed, and Leydig and Sertoli cell counts and Johnsen scores were measured. The control diet group had significant testicular oxidative stress and mild tissue deterioration. Leydig and Sertoli cell counts, tubule diameters, and Johnsen scores were significantly decreased in the exposure groups. Apricot-rich diet significantly ameliorated the oxidative status and prevented the damage in tubular histology. The protective effects were prominent when the diet was maintained throughout the time course and were partially protected when the diet was initiated after exposure. The natural antioxidant activity of apricot ameliorates the delayed detrimental effects of low-dose irradiation on testis tissue. The high total antioxidant capacity of the apricot deserves further investigation.

Effect of noise exposure on gap detection in rats

The effects of intense (110–120 dB) noise exposure (broadband noise for one hour) on temporal resolution was estimated in rats by measuring the behavioural gap detection threshold (GDT). Changes in GDT after 120 dB noise exposure were compared with changes in the threshold and amplitude of middle latency responses (MLR) recorded in response to tone stimuli. GDT values increased from 1.6 to 4.3 or 7.8 ms after exposure to 110 or 115 dB SPL, respectively; GDT recovered to pre-exposure values in 3–7 days. Three main types of noise-induced changes were observed after 120 dB SPL exposure: (I) GDT changes similar to those following noise exposure to 115 dB SPL and maximal hearing threshold shifts (TSs) at high frequencies of about 45 dB; (II) more pronounced changes in GDT (up to 60 ms) with maximal hearing threshold shifts of about 65 dB and (III) a lack of reliable responses to gap during the first weeks post-exposure with maximal hearing threshold shifts of about 80 dB. An increased GDT was present two months after noise exposure in animals with types II and III post-exposure changes; enhanced MLR amplitudes were also found in most of these in the first post-exposure week. The pronounced deficit in gap detection in some rats after 120 dB SPL noise exposure may signal the presence of a noise-induced tinnitus.

Paternal Effects from Methamidophos Administration in Mice

In this study, the mouse was used to evaluate paternal germline exposure to the organophosphate methamidophos for its potential to produce adverse effects on spermatozoa and in the offspring. There have been reports that organophosphate exposure can increase abnormal sperm morphology in mice. However, effects transmitted to the offspring following paternal exposure have not been reported previously. The maximum tolerated dose (MTD) was 7.5 mg kg−1 body weight and this dose resulted in no deaths, although blood plasma cholinesterase activity was still decreased. Males were euthanized 4 weeks after an acute intraperitoneal injection of methamidophos (0.5, 3.75, 5.0, and 7.5 mg kg−1 body wt) and the number of spermatids per gram testes and sperm morphology were analyzed. In this study, abnormal sperm morphology on a per group basis exhibited a dose–response significantly related to increased methamidophos exposure as indicated by regression analysis and a nested ANOVA (p < 0.0001). Preimplantation embryos that were conceived 6 weeks after paternal methamidophos exposure (5 mg kg−1 body wt) exhibited a significant increase in cleavage arrest. Fertility of males was also affected as shown by a decrease in the number of two- to four-cell embryos per male (postexposure week 6) and an increase in the number of degenerated embryos (postexposure weeks 4–6). We conclude that methamidophos may have the potential to produce transmissible adverse embryonic effects following an acute paternal germline exposure.

Dominant lethal studies in male mice after exposure to a 50 Hz magnetic field

The potential mutagenicity of power frequency magnetic fields was investigated using a dominant lethal assay in mice. A total of 42 male mice were exposed for 8 weeks to a 50 Hz sinusoidal magnetic field at 10 mT (rms) and 47 males acted as simultaneous cage controls. Each male was subsequently mated with two females on weeks 1, 3, 5, 7, and 9 post-exposure. The numbers of pregnant females, corpora lutea, and live and dead implants were recorded. Multiple logistic regression analyses examined the effects of exposure on pregnancy rate, pre-implantation survival and post-implantation survival. There were no statistically significant differences in overall response between exposed and control groups, nor was there any significant effect of exposure in any post-exposure week. Thus, exposure to power frequency magnetic fields at 10 mT for the approximate period of spermatogenesis did not appear to induce dominant lethal mutation in the germ cells of male mice.

Populations of Salmonella typhimurium in internal organs of experimentally infected carrier swine

Summary Experiments were conducted to determine comparative populations of Salmonella typhimurium in the most commonly infected body organs of long-term carrier swine. Naturally farrowed Salmonella-free pigs (n = 58) were orally exposed to S typhimurium when they were 47 days old. Necropsy of 3 to 5 randomly selected pigs was conducted at 3, 7, 10, 14, and 17 days and at 3, 4, 5, 6, 8, 12, 16, 20, 24, and 28 weeks after exposure. Mean populations (log10/g) of S typhimurium in palatine tonsils, ileum, cecum (wall and contents), ascending colon (wall and contents), and mandibular and ileocolic lymph nodes were estimated at each necropsy, using a most-probable-number method of bacteriologic examination. Populations of organisms in cecum and colon were similar to each other throughout the duration of the study. Mean populations (log10/g) associated with cecal and colonic walls decreased from 6.1 and 6.6, respectively, during the first postexposure (pe) week to ≤ 1.67 from pe weeks 4 to 28. Populations (log10/g) associated with cecal and colonic contents decreased from 5.6 and 5.5, respectively, at pe day 3 to 2.5 and 2.7, respectively, at pe week 4, and remained ≤ 2.8 until week 28. Populations (log10/g) associated with intestinal walls and contents were closely correlated during the study. Population (log10/g) in the ileum was ≥ 5.3 from pe days 3 to 17, then varied between 5.4 and -0.4 up to pe week 28. Population (log10/g) of S typhimurium in the tonsils varied from 6.4 to 5.5 up to pe week 5, then decreased to a range of 5.3 to 4.0 until pe week 24, and further decreased to 1.1 at pe week 28. Population (log10/g) in mandibular lymph nodes decreased from 3.5 at pe day 3 to 0.8 at week 8, then ≤ 1.0 up to week 28. In ileocolic lymph nodes, population decreased from 4.4 during the first pe week to ‒0.2 at 8 weeks, then ≤ 1.5 up to week 28. Fecal samples obtained weekly or biweekly throughout the study were 50 to 100% culture-positive (av, 85.4%). Results indicate that the long-term carrier state of S typhimurium in most infected body organs of swine exists mainly at a low, but fairly stable population.

Altered lung function in rats after subacute exposure to n-butyl isocyanate.

The objectives of this study were to use pulmonary function tests, blood gas measurements and bronchoalveolar lung lavage (BAL) to characterize lesions in the respiratory tract of young adult male Wistar rats as a result of a 5-day exposure (6 h/day) to 0, 1.1, 6.2, 15 or 26 mg n-butyl isocyanate (n-BIC)/m3 air. Further objectives were to probe the diagnostic sensitivities of these procedures in comparison with more traditional evaluations (clinical observation, lung weight, histopathology). Measurements were performed during post-exposure weeks 2 and 5. Most rats exposed to 26 mg/m3 died or were sacrificed in a moribund state during post-exposure week 2. All other rats survived the exposure regimen. In rats exposed to 15 and 26 mg/m3 a significant decrease in body weight, laboured breathing, hypoactivity, nasal discharge, cyanosis, and hypothermia were observed. Pulmonary function measurements revealed increased total lung capacity (TLC) and residual volume (RV), decreased forced expiratory flow rates and quasi-static compliance in rats exposed to 26 mg/m3. At the end of the observation period rats exposed to 6.2 and 15 mg/m3 air were hyperresponsive to an acetylcholine bronchoprovocation aerosol. Arterial blood gas measurements revealed an arterial hypoxia and an increase in venous admixture, suggesting a severe mismatch of the ventilation-perfusion relationship. Biochemical and cellular components in BAL fluid (BALF) indicated a concentration dependent and protracted increase of polymorphonuclear leucocytes and further inflammatory parameters. In the 1.1 mg/m3 group BALF parameters were not significantly elevated. The major histopathological lesions of the lung were thickening of septa, emphysema, and intra-alveolar oedema in rats exposed to 26 mg/m3.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of chronic carbon disulfide inhalation on sensory and motor function in the rat

Chronic carbon disulfide (CS 2) exposure produces debilitating motor, sensory and neuropsychiatric consequences in humans. Sensory, especially auditory, tests have been considered for indexing early intoxication. This study examines effects of chronic CS 2 exposure in rat upon auditory and neuromuscular function using reflex modulation audiometry to test the feasibility of using pure tone detection thresholds as such an index. This method is sensitive to the differential effects of toxicants upon acoustic and neuromuscular functioning. Rats were tested before, during and after five or 12 weeks of 500 ppm CS 2 inhalation, six h/day, five days/week. Neuromuscular integrity, reflected by baseline startle amplitude, decreased 50% after five weeks and 67% following 12 weeks of CS 2 exposure; recovery to 70% of preexposure values occurred on the fourth postexposure week. Twelve weeks of CS 2 inhalation had no significant effect upon acoustic thresholds. Pure tone detection thresholds, therefore, do not appear adequate to index early CS 2 exposure levels in the rat, as severe neuromuscular compromise occurred at a time when acoustic thresholds remained stable.

The fungicide methyl 2-benzimidazole carbamate causes infertility in male Sprague-Dawley rats.

A serial breeding technique was used to evaluate the fertility of male Sprague-Dawley rats after exposure to the fungicide carbendazim (methyl 2-benzimidazole carbamate). Proven-fertile male rats (90 days old) received 10 daily doses of corn oil or carbendazim (400 mg/kg/day) peroral. Each male was bred with a new female each week; breeding began on the third day of treatment and continued for 32 wk after the last day of chemical exposure. Twelve days after each breeding period, the females were killed, their uteri were examined for resorptions, and the number of dead and viable fetuses was determined. All males were killed 35 wk post exposure, and testicular tissue was prepared for histopathological examination by vascular perfusion. Fertility (percent fertile as indicated by pregnant females) of males in the carbendazim-treated group was depressed the first post-exposure week; 10 of the 24 treated males failed to produce a pregnant female as compared with no failures in the control group. By the fifth post-exposure week, 16 of the 24 carbendazim-treated males were infertile. Of these 16 males, 4 recovered fertility after a failure to produce a pregnant female for 5-11 consecutive breeding periods. However, 12 of the males did not recover fertility during the remainder of the 32-wk post-exposure period. Histological examinations of testicular sections 245 days post exposure revealed that exposure to carbendazim caused severe seminiferous tubular atrophy (greater than 85% of tubules were atrophic) in those carbendazim-treated males that failed to recover fertility.(ABSTRACT TRUNCATED AT 250 WORDS)

Role of epididymal inflammation in the induction of dominant lethal mutations in Fischer 344 rat sperm by methyl chloride.

This study assessed the possible relationship between methyl chloride (MeCl)-induced epididymal inflammation and the formation of dominant lethal mutations in sperm of Fischer 344 rats. Groups of 40 males were exposed to MeCl (3000 ppm 6 hr/day for 5 days), with or without concurrent treatment with the anti-inflammatory agent 3-amino-1-[m-(trifluoromethyl)phenyl]-2-pyrazoline (BW 755C; 10 mg/kg, i.p. 1 hr pre- and postexposure); BW 755C was shown previously to inhibit MeCl-induced epididymal inflammation. Control groups (n = 20) were either untreated, injected as described above with BW 755C, or injected on the afternoon of day 5 with triethylenemelamine (0.2 mg/kg), a known dominant lethal mutagen. Each male was caged with one female weekly for 3 weeks; 12-18 days after mating, females were killed to assess dominant lethal parameters. In females bred to MeCl-exposed males, significant increases were observed in postimplantation loss at postexposure week 1 (0.84 dead implants per female vs. 0.29 in untreated controls) and in dead implants/total implants at both week 1 (0.10 vs. 0.04 control) and week 2 (0.24 vs. 0.06 control). These increases were not observed in females bred to males treated with BW 755C during MeCl exposure. Coadministration of BW 755C to males along with MeCl also reduced the percentage of mated females with two or more postimplantation losses from 31% to 8% (week 1) and 30% to 12% (week 2). Therefore, the dominant lethal mutations induced by MeCl appear to be a consequence of its induction of inflammation in the epididymis. These data demonstrate the potential genotoxicity of inflammatory processes in vivo.

Acute inhalation studies with methyl isocyanate vapor: I. Methodology and LC50 determinations in guinea pigs, rats, and mice

Groups of male and female Fischer 344 rats, B6C3F1 mice, and Hartley guinea pigs were exposed once for 6 hr to mean concentrations of 10.5, 5.4, 2.4, 1.0, or 0 (control) ppm of methyl isocyanate (MIC) vapor. Rats and mice were also exposed to 20.4 ppm of MIC. No deaths occurred in animals exposed to 2.4 or 1.0 ppm. The majority of deaths for the 20.4- and 10.5-ppm groups occurred during postexposure Days 1 through 3, while at 5.4 ppm deaths were observed throughout the 14-day postexposure period. The 6-hr LC50 values (with 95% confidence limits) were 6.1 (4.6 to 8.2) ppm for rats, 12.2 (8.4 to 17.5) ppm for mice, and 5.4 (4.4 to 6.7) ppm for guinea pigs. Notable clinical observations during and immediately following MIC exposure were lacrimation, perinasal/perioral wetness, respiratory difficulty (e.g., mouth breathing), decreased activity, ataxia, and hypothermia. The frequency of clinical signs decreased during the second postexposure week. Body weight losses were common in all species following MIC exposures of 2.4 ppm or greater. At 1.0 ppm, only female mice had body weight depression. Recovery of body weight loss was observed in the 5.4- (guinea pigs only), 2.4-, and 1.0-ppm concentration groups. The lungs of all animals that died were discolored. Following microscopic examination of the respiratory tract, deaths were attributed to pulmonary edema and congestion. In a separate study, Fischer 344 rats and Hartley guinea pigs were exposed once for 4 hr to mean concentrations of 36.1, 25.6, 15.2, or 5.2 ppm of MIC vapor. In general, the results were similar to those of the single 6-hr exposure study.

Evaluation of propylene oxide for mutagenic activity in 3 in vivo test systems

Propylene oxide (CAS No. 75-56-9) was tested for mutagenic activity following vapor exposure using 3 in vivo test systems. Rat dominant lethal and mouse sperm-head morphology assays were conducted using males exposed to propylene oxide at 300 ppm in a dynamic exposure chamber for 7 h per day on 5 consecutive days. A sex-linked recessive lethal test in Drosophila melanogaster employed a 24-h static exposure to propylene oxide at 645 ppm. Male mice were killed 1, 3, 5, 7, and 9 weeks post-exposure for evaluation of sperm-head morphology. Propylene oxide exposure did not result in an increase in abnormal forms. Male rats were mated with 2 virgin females per week for 6 weeks following exposure. A statistically significant increase in preimplantation losses and a statistically significant reduction in the number of lving implants in the first post-exposure week did not appear to be treatment related. A highly significant increase in sex-linked recessive lethal mutations was observed in two germ cell stages (mature sperm and developing spermatocytes). These results warrant continued caution in potential human exposure to propylene oxide.

The retention and fate of inhaled plutonium dioxide in dogs.

An inhalation study of plutonium dioxide in twenty-two dogs following single exposures is described. The study emphasized the clearance of PuO2 dust from the lungs and its fate. The pulmonary clearance process, computed by several methods including in vivo counting, can be described as a bi-phasic exponential with mean biological half times of about 1 and 400 days, respectively. By excluding the first post-exposure week, the lung clearance can also be approximated by the power function t−0.22. The tissue distribution of plutonium in animals sacrificed between 16 and 468 days after exposure is described. The bronchial lymph nodes and the lungs had the highest plutonium concentration and, on the average, accounted for 95 per cent of the body burden. The principal pathological effects noted were associated with these tissues. Few extrapulmonary tissues, except the gonads and lymph nodes, appear to have received important radiation doses. The absorption of plutonium averaged 3.5 per cent of the PuO2 deposited. A disproportionate part of the absorption appeared to have occurred shortly after exposure. The report also describes the elimination kinetics of plutonium and discusses some problems in interpreting excretion data.

Mechanism of the protective action of 2-mercaptoethylguanidine (MEG) against whole-body radiation in mice.

Summary1. 2-mercaptoethylguanidine will protect mice against the lethal effects of an x-ray exposure of 900 r, converting an LD100 in 13 days to an LD10 in 30 days.2. Body-weight, spleen, thymus and small-intestine weight, total white-cell count and total marrow count (right femur) are identical for at least the first 48 hours post-exposure in mice given 900 r, whether treated with MEG or not.3. Of the parameters studied only the marrow count and small-intestine weight show definite recovery by the fifth day in MEG-treated mice; all others recover during the second post-exposure week.4. The evidence presented indicates that MEG-treated mice recover faster without lessening the initial damage due to radiation.5. A dose-reduction factor of about 2 was found, based on the known 30-day lethality of this strain of mouse.

Modification of radiation injury in rats through gastrointestinal tract shielding and bone marrow therapy.

The efficacy of postexposure administration of isologous bone marrow in preventing mortality was compared in whole-body x-irradiated rats and in animals exposed with three grossly different amounts of the gastrointestinal tract exteriorized and lead shielded. Survival was not enhanced by the therapy in animals which sustained severe intestinal radiation injury and died during the first postexposure week. However, significant protection was afforded by the treatment in animals in which the incidence of acute intestinal radiation deaths was reduced or virtually eliminated by intestinal shielding and deaths in the untreated controls occurred predominantly during the second postexposure week from bone marrow injury. The data demonstrate that bone marrow therapy is effective primarily in preventing deaths associated with radiation-induced damage to the hematopoietic tissue and not to the intestine, and hence its therapeutic value becomes limited as the prominence of intestinal injury increases with increasing radiation dosage in the whole-body irradiated animal.