RESUMO
Exposure to ionizing radiation can induce genetic aberrations via unrepaired DNA strand breaks. To investigate quantitatively the dose-effect relationship at the molecular level, we irradiated dry pBR322 plasmid DNA with 3 MeV protons and assessed fragmentation yields at different radiation doses using long-read sequencing from Oxford Nanopore Technologies. This technology applied to a reference DNA model revealed dose-dependent fragmentation, as evidenced by read length distributions, showing no discernible radiation sensitivity in specific genetic sequences. In addition, we propose a method for directly measuring the single-strand break (SSB) yield. Furthermore, through a comparative study with a collection of previous works on dry DNA irradiation, we show that the irradiation protocol leads to biases in the definition of ionizing sources. We support this scenario by discussing the size distributions of nanopore sequencing reads in the light of Geant4 and Geant4-DNA simulation toolkit predictions. We show that integrating long-read sequencing technologies with advanced Monte Carlo simulations paves a promising path toward advancing our comprehension and prediction of radiation-induced DNA fragmentation.
Assuntos
Fragmentação do DNA , Método de Monte Carlo , Plasmídeos , Plasmídeos/genética , Fragmentação do DNA/efeitos da radiação , Relação Dose-Resposta à Radiação , Análise de Sequência de DNA/métodos , Quebras de DNA de Cadeia Simples/efeitos da radiação , DNA/genéticaRESUMO
In the event of a large-scale incident involving radiological or nuclear exposures, there is a potential for large numbers of individuals to have received doses of radiation sufficient to cause adverse health effects. It is imperative to quickly identify these individuals in order to provide information to the medical community to assist in making decisions about their treatment. The cytokinesis-block micronucleus assay is a well-established method for performing biodosimetry. This assay has previously been adapted to imaging flow cytometry and has been validated as a high-throughput option for providing dose estimates in the range of 0-10â¯Gy. The goal of this study was to test the ability to further optimize the assay by reducing the time of culture to 48â¯h from 68â¯h as well as reducing the volume of blood required for the analysis to 200 µL from 2â¯mL. These modifications would provide efficiencies in time and ease of processing impacting the ability to manage large numbers of samples and provide dose estimates in a timely manner. Results demonstrated that either the blood volume or the culture time could be reduced while maintaining dose estimates with sufficient accuracy for triage analysis. Reducing both the blood volume and culture time, however, resulted in poor dose estimates. In conclusion, depending on the needs of the scenario, either culture time or the blood volume could be reduced to improve the efficiency of analysis for mass casualty scenarios.
Assuntos
Citocinese , Citometria de Fluxo , Testes para Micronúcleos , Testes para Micronúcleos/métodos , Humanos , Citometria de Fluxo/métodos , Fatores de Tempo , Volume Sanguíneo , Relação Dose-Resposta à Radiação , AnimaisRESUMO
In this paper, we demonstrated the biological effects of acute low-dose neutrons on the whole body of rats and investigated the impact of that level of neutron dose to induce an in vivo radio-adaptive response. To understand the radio-adaptive response, the examined animals were exposed to acute neutron radiation doses of 5 and 10 mSv, followed by a 50 mSv challenge dose after 14 days. After irradiation, all groups receiving single and double doses were kept in cages for one day before sampling. The electron paramagnetic resonance (EPR) method was used to estimate the radiation-induced radicals in the blood, and some hematological parameters and lipid peroxidation (MDA) were determined. A comet assay was performed beside some of the antioxidant enzymes [catalase enzyme (CAT), superoxide dismutase (SOD), and glutathione (GSH)]. Seven groups of adult male rats were classified according to their dose of neutron exposure. Measurements of all studied markers are taken one week after harvesting, except for hematological markers, within 2 h. The results indicated lower production of antioxidant enzymes (CAT by 1.18-5.83%, SOD by 1.47-17.8%, and GSH by 11.3-82.1%). Additionally, there was an increase in red cell distribution width (RDW) (from 4.61 to 25.19%) and in comet assay parameters such as Tail Length, (from 6.16 to 10.81 µm), Tail Moment, (from 1.17 to 2.46 µm), and percentage of DNA in tail length (DNA%) (from 9.58 to 17.32%) in all groups exposed to acute doses of radiation ranging from 5 to 50 mSv, respectively. This emphasizes the ascending harmful effect with the increased acute thermal neutron doses. The values of the introduced factor of radio adaptive response for all markers under study reveal that the lower priming dose promotes a higher adaptation response and vice versa. Ultimately, the results indicate significant variations in DNA%, SOD enzyme levels, EPR intensity, total Hb concentration, and RDWs, suggesting their potential use as biomarkers for acute thermal neutron dosimetry. Further research is necessary to validate these measurements as biodosimetry for radiation exposure, including investigations involving the response impact of RAR with varied challenge doses and post-irradiation behavior.
Assuntos
Biomarcadores , Nêutrons , Animais , Ratos , Masculino , Biomarcadores/metabolismo , Superóxido Dismutase/metabolismo , Peroxidação de Lipídeos/efeitos da radiação , Radiometria/métodos , Relação Dose-Resposta à Radiação , Dano ao DNA/efeitos da radiação , Adaptação Fisiológica/efeitos da radiação , Catalase/metabolismo , Glutationa/metabolismo , Glutationa/sangue , Ensaio Cometa , Estresse Oxidativo/efeitos da radiação , Espectroscopia de Ressonância de Spin Eletrônica/métodosRESUMO
BACKGROUND AND PURPOSE: Cone beam computed tomography (CBCT) is routinely used in radiotherapy to localize target volume. The aim of our study was to determine the biological effects of CBCT dose compared to subsequent therapeutic dose by using in vitro chromosome dosimetry. MATERIALS AND METHODS: Peripheral blood samples from five healthy volunteers were irradiated in two phantoms (water filled in-house made cylindrical, and Pure Image CTDI phantoms) with 6 MV FFF X-ray photons, the dose rate was 800 MU/min and the absorbed doses ranged from 0.5 to 8 Gy. Irradiation was performed with a 6 MV linear accelerator (LINAC) to generate a dose-response calibration curve. In the first part of the investigation, 1-5 CBCT imaging was used, in the second, only 2 Gy doses were delivered with a LINAC, and then, in the third part, a combination of CBCT and 2 Gy irradiation was performed mimicking online adapted radiotherapy treatment. Metaphases were prepared from lymphocyte cultures, using standard cytogenetic techniques, and chromosomal aberrations were evaluated. Estimate doses were calculated from chromosome aberrations using dose-response curves. RESULTS: Samples exposed to X-ray from CBCT imaging prior to treatment exhibited higher chromosomal aberrations and Estimate dose than the 2 Gy therapeutic (real) dose, and the magnitude of the increase depended on the number of CBCTs: 1-5 CBCT corresponded to 0.04-0.92 Gy, 1 CBCT + 2 Gy to 2.32 Gy, and 5 CBCTs + 2 Gy to 3.5 Gy. CONCLUSION: The estimated dose based on chromosomal aberrations is 24.8% higher than the physical dose, for the combination of 3 CBCTs and the therapeutic 2 Gy dose, which should be taken into account when calculating the total therapeutic dose that could increase the risk of a second cancer. The clinical implications of the combined radiation effect may require further investigation.
Assuntos
Aberrações Cromossômicas , Tomografia Computadorizada de Feixe Cônico , Linfócitos , Imagens de Fantasmas , Dosagem Radioterapêutica , Humanos , Tomografia Computadorizada de Feixe Cônico/métodos , Aberrações Cromossômicas/efeitos da radiação , Linfócitos/efeitos da radiação , Raios X , Relação Dose-Resposta à Radiação , Radiometria/métodosRESUMO
The effects of low-dose radiation exposure remain a controversial topic in radiation biology. This study compares early (0.5, 4, 24, 48, and 72 h) and late (5, 10, and 15 cell passages) post-irradiation changes in γH2AX, 53BP1, pATM, and p-p53 (Ser-15) foci, proliferation, autophagy, and senescence in primary fibroblasts exposed to 100 and 2000 mGy X-ray radiation. The results show that exposure to 100 mGy significantly increased γH2AX, 53BP1, and pATM foci only at 0.5 and 4 h post irradiation. There were no changes in p-p53 (Ser-15) foci, proliferation, autophagy, or senescence up to 15 passages post irradiation at the low dose.
Assuntos
Autofagia , Proliferação de Células , Senescência Celular , Reparo do DNA , Fibroblastos , Humanos , Fibroblastos/efeitos da radiação , Fibroblastos/metabolismo , Autofagia/efeitos da radiação , Senescência Celular/efeitos da radiação , Reparo do DNA/efeitos da radiação , Raios X/efeitos adversos , Proliferação de Células/efeitos da radiação , Proteína 1 de Ligação à Proteína Supressora de Tumor p53/metabolismo , Histonas/metabolismo , Relação Dose-Resposta à Radiação , Proteína Supressora de Tumor p53/metabolismo , Proteína Supressora de Tumor p53/genética , Células Cultivadas , Dano ao DNA/efeitos da radiaçãoRESUMO
This study explores the impact of densely-ionizing radiation on non-cancer and cancer diseases, focusing on dose, fractionation, age, and sex effects. Using historical mortality data from approximately 21,000 mice exposed to fission neutrons, we employed random survival forest (RSF), a powerful machine learning algorithm accommodating nonlinear dependencies and interactions, treating cancer and non-cancer outcomes as competing risks. Unlike traditional parametric models, RSF avoids strict assumptions and captures complex data relationships through decision tree ensembles. SHAP (SHapley Additive exPlanations) values and variable importance scores were employed for interpretation. The findings revealed clear dose-response trends, with cancer being the predominant cause of mortality. SHAP value dose-response shapes differed, showing saturation for cancer hazard at high doses (> 2 Gy) and a more linear pattern at lower doses. Non-cancer responses remained more linear throughout the entire dose range. There was a potential inverse dose rate effect for cancer, while the evidence for non-cancer was less conclusive. Sex and age effects were less pronounced. This investigation, utilizing machine learning, enhances our understanding of the patterns of non-cancer and cancer mortality induced by densely-ionizing radiations, emphasizing the importance of such approaches in radiation research, including space travel and radioprotection.
Assuntos
Aprendizado de Máquina , Nêutrons , Animais , Camundongos , Feminino , Masculino , Relação Dose-Resposta à Radiação , Fatores Etários , Fatores Sexuais , Neoplasias Induzidas por Radiação/mortalidade , Fracionamento da Dose de RadiaçãoRESUMO
BACKGROUND: Sarcocystis is a food-borne zoonotic protozoan whose final hosts are humans, dogs, cats, and other carnivores and intermediate hosts are birds and mammals, especially humans and herbivores. Humans become infected by eating raw and undercooked meat contaminated with bradyzoites or by consuming water or food contaminated with the sporocyst stage of the parasite. OBJECTIVES: The aim of this study was to investigate the effects of gamma radiation and electron beam on the survival rate of Sarcocystis bradyzoites in infected beef and to determine the effective dose. METHODS: Three replicates of 100 g of infected meat were treated with different doses (0.5, 1, 1.5 and 2 kGy). As a control, 20 g of contaminated meat was stored separately at 4°C. The viability of the bradyzoites after digestion in pepsin solution was assessed, stained (trypan blue) and unstained, under a stereomicroscope. To assess survival of the bradyzoites, the irradiated meat samples were fed to 30 dogs. After 10 days, faecal samples were examined for sporocysts. RESULTS: The results showed that the highest and lowest mortality rate of Sarcocystis bradyzoites in infected organs using electron beam at a dose of 2 kGy were 92.5% and 100%, respectively, and the lowest mortality rate at a dose of 0.5 kGy were 2.5% and 7.89%, respectively. CONCLUSION: The results of statistical analysis showed that the mortality rate of Sarcocystis bradyzoites was significant between different doses of gamma ray and electron beam, so that gamma rays were better compared to electron beam in destroying Sarcocystis bradyzoites.
Assuntos
Sarcocystis , Sarcocystis/efeitos da radiação , Sarcocystis/fisiologia , Animais , Bovinos , Sarcocistose/veterinária , Sarcocistose/parasitologia , Carne Vermelha/parasitologia , Raios gama , Cães , Irradiação de Alimentos , Relação Dose-Resposta à Radiação , Doenças dos Bovinos/parasitologia , ElétronsRESUMO
Various biological effects of ionizing radiation, especially continuous exposure to low-dose radiation (LDR), have attracted considerable attention. Impaired bone structure caused by LDR has been reported, but little is known about the mechanism involved in the disruption of bone metabolism. In this study, given that LDR was found to (at a cumulative dose of 0.10â¯Gy) disturb the serum Mg2+ level and Notch1 signal in the mouse femur tissues, the effects of LDR on osteogenesis and the underlying molecular mechanisms were investigated based on an in vitro culture system for bone marrow stromal cells (BMSCs). Our data showed that cumulative LDR suppressed the osteogenic potential in BMSCs as a result of upregulation of Notch1 signaling. Further analyses indicated that the upregulation of NICD1 (Notch1 intracellular domain), the key intracellular domain for Notch1 signaling, under LDR was a consequence of enhanced protein stabilization caused by SUMOylation (small ubiquitin-like modification). Specifically, the downregulation of SENP1 (sentrin/SUMO-specific protease 1) expression induced by LDR enhanced the SUMOylation of NICD1, causing the accumulation of Notch1 signaling, which eventually inhibited the osteogenic potential of BMSCs. In conclusion, this work expounded on the mechanisms underlying the impacts of LDR on bone metabolism and shed light on the research on bone regeneration under radiation.
Assuntos
Diferenciação Celular , Células-Tronco Mesenquimais , Osteogênese , Receptor Notch1 , Sumoilação , Animais , Osteogênese/efeitos da radiação , Camundongos , Sumoilação/efeitos da radiação , Receptor Notch1/metabolismo , Receptor Notch1/genética , Células-Tronco Mesenquimais/efeitos da radiação , Diferenciação Celular/efeitos da radiação , Transdução de Sinais/efeitos da radiação , Masculino , Fêmur/efeitos da radiação , Relação Dose-Resposta à RadiaçãoRESUMO
Numerous dose rate effects have been described over the past 6-7 decades in the radiation biology and radiation oncology literature depending on the dose rate range being discussed. This review focuses on the impact and understanding of altering dose rates in the context of radiation therapy, but does not discuss dose rate effects as relevant to radiation protection. The review starts with a short historic review of early studies on dose rate effects, considers mechanisms thought to underlie dose rate dependencies, then discusses some current issues in clinical findings with altered dose rates, the importance of dose rate in brachytherapy, and the current timely topic of the use of very high dose rates, so-called FLASH radiotherapy. The discussion includes dose rate effects in vitro in cultured cells, in in vivo experimental systems and in the clinic, including both tumors and normal tissues. Gaps in understanding dose rate effects are identified, as are opportunities for improving clinical use of dose rate modulation.
Assuntos
Relação Dose-Resposta à Radiação , Humanos , Animais , História do Século XX , Braquiterapia/história , Braquiterapia/métodos , Dosagem Radioterapêutica , Neoplasias/radioterapia , História do Século XXI , Radiobiologia/históriaRESUMO
PURPOSE: Nuclear applications are being increasingly used in various fields, necessitating studies to protect from radiation hazards and their effects. In this study, five different chemical structures of pyrazolo [3,4-b] pyridine derivatives were synthesized. The gamma and neutron radiation protective abilities of these samples were determined and demonstrated their potential use as ingredients in radioprotective drugs. MATERIAL AND METHODS: Gamma radiation absorption parameters were calculated both theoretical and experimental. Important attenuation parameters for fast neutrons (4.5 MeV energy radiation) were figured out using the Monte Carlo simulation Geant4 code. Additionally, experimental dose rates were measured for each sample and compared to those of Paraffin and high-density polyethylene, an organic substance. Besides, Ames/Salmonella test system was aimed to detecting genotoxicity features of pyrazolo pyridine derivatives. RESULTS: All results demonstrated that each sample possesses both gamma and neutron radiation attenuation capabilities. It was determined that sample PPC4 (C20H14BrN5) exhibited the highest gamma radiation attenuation capacity among all samples, while sample PPC2 (C22H20N6) displayed an excellent neutron stopping capacity. The genotoxic properties of pyrazolo[3,4-b] pyridine derivatives were examined using the Ames/Salmonella test, and as a result, it was determined that these substances did not exhibit genotoxic effects at test doses up to 5 mM. CONCLUSION: All obtained results indicate that all PPC (pyrazolo[3,4-b] pyridine derivatives) samples do not possess a toxic effect, and they can be utilized as an active substance for the development of a drug or cream with protective properties against both gamma and neutron radiations.
Assuntos
Raios gama , Nêutrons , Pirazóis , Piridinas , Protetores contra Radiação , Piridinas/química , Pirazóis/farmacologia , Pirazóis/química , Pirazóis/toxicidade , Protetores contra Radiação/farmacologia , Protetores contra Radiação/química , Testes de Mutagenicidade , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/efeitos da radiação , Relação Dose-Resposta à Radiação , Método de Monte CarloRESUMO
Radiotherapy in the head-and-neck area is one of the main curative treatment options. However, this comes at the cost of varying levels of normal tissue toxicity, affecting up to 80% of patients. Mucositis can cause pain, weight loss and treatment delays, leading to worse outcomes and a decreased quality of life. Therefore, there is an urgent need for an approach to predicting normal mucosal responses in patients prior to treatment. We here describe an assay to detect irradiation responses in healthy oral mucosa tissue. Mucosa specimens from the oral cavity were obtained after surgical resection, cut into thin slices, irradiated and cultured for three days. Seven samples were irradiated with X-ray, and three additional samples were irradiated with both X-ray and protons. Healthy oral mucosa tissue slices maintained normal morphology and viability for three days. We measured a dose-dependent response to X-ray irradiation and compared X-ray and proton irradiation in the same mucosa sample using standardized automated image analysis. Furthermore, increased levels of inflammation-inducing factors-major drivers of mucositis development-could be detected after irradiation. This model can be utilized for investigating mechanistic aspects of mucositis development and can be developed into an assay to predict radiation-induced toxicity in normal mucosa.
Assuntos
Mucosa Bucal , Humanos , Mucosa Bucal/efeitos da radiação , Raios X/efeitos adversos , Lesões por Radiação/etiologia , Lesões por Radiação/patologia , Masculino , Mucosite/etiologia , Mucosite/patologia , Feminino , Relação Dose-Resposta à Radiação , Estomatite/etiologia , Estomatite/patologia , Adulto , Pessoa de Meia-IdadeRESUMO
Recurrent computed tomography (CT) examination has become a common diagnostic procedure for several diseases and injuries. Though each singular CT scan exposes individuals at low doses of low linear energy transfer (LET) radiation, the cumulative dose received from recurrent CT scans poses an increasing concern for potential health risks. Here, we evaluated the biological effects of recurrent CT scans on the DNA damage response (DDR) in human fibroblasts and retinal pigment epithelial cells maintained in culture for five months and subjected to four CT scans, one every four weeks. DDR kinetics and eventual accumulation of persistent-radiation-induced foci (P-RIF) were assessed by combined immunofluorescence for γH2AX and 53BP1, i.e., γH2AX/53BP1 foci. We found that CT scan repetitions significantly increased both the number and size of γH2AX/53BP1 foci. In particular, after the third CT scan, we observed the appearance of giant foci that might result from the overlapping of individual small foci and that do not associate with irreversible growth arrest, as shown by DNA replication in the foci-carrying cells. Whether these giant foci represent coalescence of unrepaired DNA damage as reported following single exposition to high doses of high LET radiation is still unclear. However, morphologically, these giant foci resemble the recently described compartmentalization of damaged DNA that should facilitate the repair of DNA double-strand breaks but also increase the risk of chromosomal translocations. Overall, these results indicate that for a correct evaluation of the damage following recurrent CT examinations, it is necessary to consider the size and composition of the foci in addition to their number.
Assuntos
Dano ao DNA , Fibroblastos , Histonas , Tomografia Computadorizada por Raios X , Proteína 1 de Ligação à Proteína Supressora de Tumor p53 , Humanos , Proteína 1 de Ligação à Proteína Supressora de Tumor p53/metabolismo , Tomografia Computadorizada por Raios X/métodos , Histonas/metabolismo , Fibroblastos/efeitos da radiação , Fibroblastos/metabolismo , Relação Dose-Resposta à Radiação , Epitélio Pigmentado da Retina/efeitos da radiação , Epitélio Pigmentado da Retina/metabolismo , Epitélio Pigmentado da Retina/diagnóstico por imagem , Epitélio Pigmentado da Retina/citologia , Linhagem Celular , Reparo do DNA , Transferência Linear de EnergiaRESUMO
PURPOSE: Radiation-induced alterations in gene expression show great promise for dose reconstruction and for severity prediction of acute health effects. Among several genes explored as potential biomarkers, FDXR is widely used due to high upregulation in white blood cells following radiation exposure. Nonetheless, the absence of a standardized protocols for gene expression-based biodosimetry is a notable gap that warrants attention to enhance the accuracy, reproducibility and reliability. The objective of this study was to evaluate the sensitivity of transcriptional biodosimetry to differences in protocols used by different laboratories and establish guidelines for the calculation of calibration curve using FDXR expression data. MATERIAL AND METHODS: Two sets of irradiated blood samples generated during RENEB exercise were used. The first included samples irradiated with known doses including: 0, 0.25, 0.5, 1, 2, 3 and 4 Gy. The second set consisted of three 'blind' samples irradiated with 1.8 Gy, 0.4 Gy and a sham-irradiated sample. After irradiation, samples were incubated at 37 °C over 24 h and sent to participating laboratories, where RNA isolation and FDXR expression analysis by qPCR were performed using sets of primers/probes and reference genes specific for each laboratory. Calibration curves based on FDXR expression data were generated using non-linear and linear regression and used for dose estimation of 'blind' samples. RESULTS: Dose estimates for sham-irradiated sample (0.020-0.024 Gy) and sample irradiated with 0.4 Gy (0.369-0.381 Gy) showed remarkable consistency across all laboratories, closely approximating the true doses regardless variation in primers/probes and reference genes used. For sample irradiated with 1.8 Gy the dose estimates were less precise (1.198-2.011 Gy) but remained within an acceptable margin for triage within the context of high dose range. CONCLUSION: Methodological differences in reference genes and primers/probes used for FDXR expression measurement do not have a significant impact on the dose estimates generated, provided that all reference genes performed as expected and the primers/probes target a similar set of transcript variants. The preferred method for constructing a calibration curve based on FDXR expression data involves employing linear regression to establish a function that describes the relationship between the logarithm of absorbed dose and FDXR ΔCt values. However, one should be careful with using non-irradiated sample data as these cannot be accurately represented on a logarithmic scale. A standard curve generated using this approach can give reliable dose estimations in a dose range from 50 mGy to 4 Gy at least.
Assuntos
Doses de Radiação , Radiometria , Calibragem , Humanos , Radiometria/métodos , Relação Dose-Resposta à Radiação , Masculino , Reprodutibilidade dos TestesRESUMO
PURPOSE: In order to achieve mutations with enhanced economic, productive, and nutritional characteristics in the two Egyptian cowpea varieties, Dokki 331 and Kaha 1, the application of gamma irradiation at different doses is employed. Additionally, this method aids in distinguishing between these mutations using simple sequence repeat (SSR) analysis. MATERIALS AND METHODS: Two different cowpea cultivars were subjected to varying doses of gamma radiation ranging from 50 to 300 Gy. In order to analyze the effects of radiation, both unirradiated and irradiated seeds from both cultivars were planted using a randomized complete block design. This experiment was conducted over a span of six generations, namely M1, M2, M3, M4, M5, and M6, starting from April 2017 and continuing until 2022. Among the various radiation doses, the cultivar Kaha 1 produced promising traits when exposed to a dose of 150 Gy, while the cultivar Dokki 331 showed favorable traits when exposed to a dose of 300 Gy. These traits were further cultivated and studied until the M6 generation. RESULTS: Induced mutations in two Egyptian cowpea varieties, Kaha 1 and Dokki 331, are subjected to varying doses of gamma radiation (0, 50, 100, 150, 200, 250, and 300 Gy). Morphological and genetic variations were observed, with mutations being induced at doses of 150 Gy for Kaha 1 and 300 Gy for Dokki 331. The mutation in Kaha 1 (beam 1) resulted in dwarfism, altered leaf shape, early flowering, increased peduncles, pods, and pod seed numbers, ultimately leading to enhanced seed production and acreage productivity. In Dokki 331, the mutations primarily affected pod color, resulting in greenish-brown pods with mosaic seeds, segregating black and gray seeds from the mosaic ones. These mutations led to an increase in the nutritional value of the seeds, including higher nitrogen content, total free amino acids, crude protein, total carbohydrates, and total sugars. The genetic diversity of the seven cowpea mutations was assessed using 20 microsatellite markers. The analysis revealed a total of 60 alleles, with an average of three alleles per locus. The allele frequency ranged from 0.2857 to 1.0, with an average of 0.6036. Gene diversity varied from 0.0 to 0.8163, while the heterozygosity was mostly zero, except for one primer (VM 37) with an average of 0.0071. The polymorphic information content (PIC) ranged from 0.7913 to 0.0, with an average of 0.4323. The Marker Index value ranged from 0.36 to 0.0, with an average of 0.152. Overall, our findings demonstrate the successful induction of mutations in Egyptian cowpea varieties using gamma rays, resulting in improved yield characteristics and nutritional value. CONCLUSIONS: Radiation as a physical mutagen is highly regarded for its effectiveness, affordability, speed, and safety in inducing mutations. Utilizing gamma rays, we successfully derived a novel cowpea variety called beam 1 mutation, which has gained approval from the Egyptian Ministry of Agriculture.
Assuntos
Raios gama , Repetições de Microssatélites , Mutação , Valor Nutritivo , Vigna , Vigna/genética , Vigna/efeitos da radiação , Vigna/crescimento & desenvolvimento , Mutação/efeitos da radiação , Egito , Relação Dose-Resposta à RadiaçãoRESUMO
INTRODUCTION: For Hiroshima and Nagasaki survivors, it has not been possible to calculate individual doses from the cytogenetic data and compare them with the physically estimated doses. This is because the cytogenetic studies used solid Giemsa staining which only provides the percent of cells bearing at least one stable-type aberration (most of the unstable-type aberrations had already disappeared), and a gamma-ray dose plus a 10-times neutron dose was used to integrate the data for both cities. OBJECTIVES: To compare the FISH-derived gamma-ray dose with the DS02R1-derived gamma-ray dose after correcting for a contribution of the neutron dose. It was also an attempt to determine if the frequency of stable-type aberrations had remained unchanged after the exposure. METHODS: Stable exchange-type aberration data was obtained using the 2-color FISH method from 1,868 atomic bomb survivors in Hiroshima and Nagasaki. The collected frequency was first extended to a genome-equivalent frequency. Then, by using known induction rates of exchange-type aberrations in vitro caused by neutrons and gamma-rays, respectively, and the mean relationship between the neutron and gamma-ray doses in the DS02R1 estimates for the survivors, the gamma-ray effect was estimated from the total yield of translocations. RESULTS: It was found that over 95% of individual cytogenetic gamma-ray doses fell within the expected range of plus/minus about 1 Gy from the DS02R1 dose and the mean slope for the linear regression was 0.98, which reassures us of the validity of the DS02R1 study. CONCLUSIONS: The present results demonstrate the validity of the individual DS02R1 doses, and that the frequency of stable-type aberrations in blood lymphocytes did not decay over the years, and thus is useful for retrospective dose evaluations of exposures which took place in the distant past.
Assuntos
Sobreviventes de Bombas Atômicas , Raios gama , Hibridização in Situ Fluorescente , Doses de Radiação , Humanos , Japão , Aberrações Cromossômicas/efeitos da radiação , Masculino , Adulto , Feminino , Relação Dose-Resposta à Radiação , Pessoa de Meia-Idade , Nêutrons , Análise Citogenética , Guerra Nuclear , SobreviventesRESUMO
High dose radiation exposures are rare. However, medical management of such incidents is crucial due to mortality and tissue injury risks. Rapid radiation biodosimetry of high dose accidental exposures is highly challenging, considering that they usually involve non uniform fields leading to partial body exposures. The gold standard, dicentric assay and other conventional methods have limited application in such scenarios. As an alternative, we propose Premature Chromosome Condensation combined with Fluorescent In-situ Hybridization (G0-PCC-FISH) as a promising tool for partial body exposure biodosimetry. In the present study, partial body exposures were simulated ex-vivo by mixing of uniformly exposed blood with unexposed blood in varying proportions. After G0-PCC-FISH, Dolphin's approach with background correction was used to provide partial body exposure dose estimates and these were compared with those obtained from conventional dicentric assay and G0-PCC-Fragment assay (conventional G0-PCC). Dispersion analysis of aberrations from partial body exposures was carried out and compared with that of whole-body exposures. The latter was inferred from a multi-donor, wide dose range calibration curve, a-priori established for whole-body exposures. With the dispersion analysis, novel multi-parametric methodology for discerning the partial body exposure from whole body exposure and accurate dose estimation has been formulated and elucidated with the help of an example. Dose and proportion dependent reduction in sensitivity and dose estimation accuracy was observed for Dicentric assay, but not in the two PCC methods. G0-PCC-FISH was found to be most accurate for the dose estimation. G0-PCC-FISH has potential to overcome the shortcomings of current available methods and can provide rapid, accurate dose estimation of partial body and high dose accidental exposures. Biological dose estimation can be useful to predict progression of disease manifestation and can help in pre-planning of appropriate & timely medical intervention.
Assuntos
Hibridização in Situ Fluorescente , Hibridização in Situ Fluorescente/métodos , Humanos , Aberrações Cromossômicas/efeitos da radiação , Exposição à Radiação/efeitos adversos , Radiometria/métodos , Doses de Radiação , Masculino , Relação Dose-Resposta à RadiaçãoRESUMO
Purpose: The purpose of this study was to analyze the extent of DNA breaks in primary uveal melanoma (UM) with regard to radiotherapy dose delivery (single-dose versus fractionated) and monosomy 3 status. Methods: A total of 54 patients with UM were included. Stereotactic radiotherapy (SRT) was performed in 23 patients, with 8 undergoing single-dose SRT (sdSRT) treatment and 15 receiving fractionated SRT (fSRT). DNA breaks in the enucleated or endoresected tumors were visualized by a TUNEL assay and quantified by measuring the TUNEL-positive area. Protein expression was analyzed by immunohistochemistry. Co-detection of chromosome 3 with proteins was performed by immuno-fluorescent in situ hybridization. Results: The amount of DNA breaks in the total irradiated group was increased by 2.7-fold (P < 0.001) compared to non-irradiated tissue. Tumors treated with fSRT were affected more severely, showing 2.1-fold more DNA damage (P = 0.007) compared to the cases after single (high) dose irradiation (sdSRT). Monosomy 3 tumors showed less DNA breaks compared to disomy 3 samples (P = 0.004). The presence of metastases after radiotherapy correlated with monosomy 3 and less DNA breaks compared to patients with non-metastatic cancer in the combined group with fSRT and sdSRT (P < 0.05). Conclusions: Fractionated irradiation led to more DNA damage than single-dose treatment in primary UM. As tumors with monosomy 3 showed less DNA breaks than those with disomy 3, this may indicate that they are less radiosensitive, which may influence the efficacy of irradiation.
Assuntos
Cromossomos Humanos Par 3 , Dano ao DNA , Melanoma , Neoplasias Uveais , Humanos , Neoplasias Uveais/radioterapia , Neoplasias Uveais/genética , Melanoma/radioterapia , Melanoma/genética , Feminino , Cromossomos Humanos Par 3/genética , Masculino , Pessoa de Meia-Idade , Idoso , Adulto , Idoso de 80 Anos ou mais , Hibridização in Situ Fluorescente , Marcação In Situ das Extremidades Cortadas , Dosagem Radioterapêutica , Imuno-Histoquímica , Radiocirurgia/efeitos adversos , Radiocirurgia/métodos , Relação Dose-Resposta à RadiaçãoRESUMO
AIMS: This study proposes to investigate the effects of microwave radiation and its thermal effects, compared to thermal effects alone, on the bioenergetics of mitochondria isolated from mouse liver. METHODS: The main parameters investigated in this study are mitochondrial respiration (coupled states: S3 and S4; uncoupled state), using a high-resolution respirometer, and swelling, using a spectrophotometer. RESULTS: Mitochondria irradiated at 2.45 GHz microwave with doses 0.085, 0.113 and 0.141 kJ/g, presented a decrease in S3 and uncoupled state, but an increase in S4. Conversely, mitochondria thermally treated at 40, 44 and 50 °C presented an increasing in S3 and S4, while uncoupled state was unaltered. Mitochondrial swelling increases as a function of the dose or temperature, indicating membrane damages in both cases. CONCLUSION: Microwave radiation and thermal effect alone indicated different bioenergetics mitochondria response. These results imply that the effects due to microwave in medical treatment are not exclusively due to the increase in temperature, but a combination of electromagnetic and thermal effects.
Assuntos
Metabolismo Energético , Micro-Ondas , Mitocôndrias Hepáticas , Animais , Camundongos , Metabolismo Energético/efeitos da radiação , Mitocôndrias Hepáticas/efeitos da radiação , Mitocôndrias Hepáticas/metabolismo , Masculino , Relação Dose-Resposta à Radiação , Temperatura , Dilatação Mitocondrial/efeitos da radiação , Respiração Celular/efeitos da radiaçãoRESUMO
PURPOSE: Stevia rebaudiana Bertoni is a perennial herb, widely used as a natural sweetener around the globe. The key compounds responsible for its sweetness includes stevioside and rebaudioside-A. In order to improve these steviol glycosides, the present study was initiated to study the effect of induced mutagenesis on growth parameters, steviol glycosides and nuclear DNA content in Stevia rebaudiana Bertoni using ten doses of gamma-rays (5-100 kR). MATERIALS AND METHODS: Healthy seeds of 'Madhuguna' variety of Stevia rebaudiana Bertoni developed and maintained at stevia breeding farm, Agrotechnology division, CSIR-Institute of Himalayan Bioresource Technology, Palampur (HP), India were irradiated with ten doses of gamma rays (600 seeds each/dose) ranging from 5 kR to 100 kR (i.e., 5, 10, 15, 20, 30, 40, 50, 60, 80 and 100 kR) using Co60 gamma irradiation chamber at CCS Haryana Agricultural University, Hisar, (Haryana), India. RESULTS: Significant variations were recorded for all the seedling traits studied while major impact was noticed on the seedling after reaching the cotyledonary stage and doses above 40 kR showed absolute mortality of the seedlings. Based on probit analysis, the optimum LD50 dose lies in the range of 20-23 kR. Glycosidic profiling of 296 mutants using high-performance liquid chromatography showed decreased total steviol glycoside content with increased radiation dose. Doses 5 kR and 10 kR, were found to be effective in increasing the overall glycosidic content. A total of 72 promising mutants were also screened for increased rebaudioside-A stevioside ratio. Comparison of nuclear DNA content using flow cytometry revealed a similar decrease in the total nuclear DNA content with increase in dosage of gamma rays. The average genome size at 5, 10, 15, 20 and 30 kR treatments were 2.72, 2.69, 2.68, 2.70 and 2.66 pg as compared to 2.72 pg in control. CONCLUSIONS: Mild dose of gamma rays (5 and 10 kR) in stevia were found to be effective in improving the mean steviol glycoside content and may be used in future stevia mutation programmes.
Assuntos
Diterpenos do Tipo Caurano , Raios gama , Stevia , Stevia/efeitos da radiação , Tolerância a Radiação , Glucosídeos , Relação Dose-Resposta à RadiaçãoRESUMO
Application of laser-generated electron beams in radiotherapy is a recent development. Accordingly, mechanisms of biological response to radiation damage need to be investigated. In this study, telomere length (TL) as endpoint of genetic damage was analyzed in human blood cells (leukocytes) and K562 leukemic cells irradiated with laser-generated ultrashort electron beam. Metaphases and interphases were analyzed in quantitative fluorescence in situ hybridization (Q-FISH) to assess TL. TLs were shortened compared to non-irradiated controls in both settings (metaphase and interphase) after irradiation with 0.5, 1.5, and 3.0 Gy in blood leukocytes. Radiation also caused a significant TL shortening detectable in the interphase of K562 cells. Overall, a negative correlation between TL and radiation doses was observed in normal and leukemic cells in a dose-dependent manner. K562 cells were more sensitive than normal blood cells to increasing doses of ultrashort electron beam radiation. As telomere shortening leads to genome instability and cell death, the results obtained confirm the suitability of this biomarker for assessing genotoxic effects of accelerated electrons for their further use in radiation therapy. Observed differences in TL shortening between normal and K562 cells provide an opportunity for further development of optimal radiation parameters to reduce side effects in normal cells during radiotherapy.