Gamma radiation induced changes in expression of heat shock proteins (Hsc70 and Hsp83) in the dengue vector Aedes aegypti (L.)

We aimed to assess the effect of gamma radiation on the expression of heat shock proteins Hsc70 and Hsp83 in Aedes aegypti. Adult males were irradiated with 50Gy of gamma radiation, and changes in the expression of proteins in SDS-PAGE gel bands corresponding to molecular weights ~60–75kDa and ~80–95kDa were analyzed at two different time points 6 and 12-hour post-irradiation, using a temporal mass spectrometry based semi-quantitative analysis. A 2-3-fold increase was observed in both proteins Hsc70 and Hsp83, at both time points. In addition, the experiment also revealed the overexpression of several other molecules such as Arginine Kinase - known to be upregulated in certain insects during stress, Esterase B1- implicated in insecticide resistance, and also down-regulation of the 26S proteasome non-ATPase regulatory subunit 1 and ubiquitin-activating enzyme E1 - both known to be involved in ubiquitin-mediated protein degradation. The results taken together with existing data on Hsp83 and Hsc70, indicate that these proteins may enhance the survival of Ae. aegypti following gamma radiation and could serve as molecular markers for the detection of radiation-induced stress.

Nanogels as prospective Biomaterial: Radio-induced Synthesis, Characterization, and Biological Assays / Los nanogeles como biomateriales prospectivos: síntesis radioinducida, caracterización y ensayos biológicos

Nowadays, there is a growing interest in biodegradable polymers-based materials due to their diverse application in the biomedical field. Most studied systems involve biocompatible micro and nanodevices, such as liposomes, dendrimer, micelles or polymeric nanogels. The use of Radiation Technology, specifically gamma radiation, to produce micro and nanogels raises the possibility to obtain higher purity products, an important feature for biomedical and pharmaceutical applications. The radio-induced synthesis, characterization, cytotoxicity evaluation, and immunological response of nanogels are described in this study. Nanogel synthesis was performed in the absence of oxygen using aqueous polyvinylpyrrolidone solutions. Crosslinking reactions were carried out at 25 °C in a gamma irradiation chamber with a 60Co source. Nanogels properties were analysed by Scanning Electron Microscopy, Attenuated Total Reflection-Fourier Transform Spectroscopy, Dynamic Light Scattering, and Viscosimetry. The cytotoxicity and immunological response were evaluated by MTT test and analysis of the neutrophil respiratory burst. The results showed that nanogels formation strongly depends on the total absorbed dose. The nanogels have an elliptical shape and their chemical structure is similar to the initial polymer. The nanogels are biocompatible and promote a low-intensity neutrophil activation, similar to the well-characterized biomaterial TiO2, suggesting their potential biomedical uses(AU)

En la actualidad existe un interés creciente en los materiales biodegradables basados en polímeros, debido a sus diversas aplicaciones en la esfera de la biomedicina. En la mayoría de los sistemas estudiados participan micro- y nanodispositivos biocompatibles, tales como liposomas, dendrímeros, micelas o nanogeles poliméricos. El uso de la tecnología de radiaciones, en particular de radiaciones gamma, para producir micro- y nanogeles, eleva la posibilidad de obtener productos de mayor pureza, un rasgo importante con vistas a su aplicación biomédica y farmacéutica. El estudio describe la síntesis radioinducida, caracterización, evaluación de la citotoxicidad y respuesta inmunológica de los nanogeles. La síntesis de los nanogeles se realizó en ausencia de oxígeno, usando soluciones acuosas de polivinilpirrolidona. Las reacciones de entrecruzamiento se realizaron a 25 ºC en cámara de irradiación gamma con una fuente de 60Co. Las propiedades de los nanogeles se analizaron mediante microscopía electrónica de barrido, espectroscopia por transformada de Fourier total atenuada, dispersión dinámica de luz y viscosimetría. La citotoxicidad y la respuesta inmunológica se evaluaron mediante prueba MTT y análisis del estallido respiratorio de neutrófilos. Los resultados muestran que la formación de nanogeles depende en gran medida de la dosis total absorbida. Los nanogeles tienen forma elíptica y su estructura química es similar a la del polímero inicial. Los nanogeles son biocompatibles y promueven una activación de neutrófilos de baja intensidad similar al bien caracterizado material TiO2, lo que sugiere usos biomédicos potenciales(AU)

Physicochemical and Tissue Response of PLA Nanofiber Scaffolds Sterilized by Different Techniques / Evaluación fisicoquímica y respuesta tisular de andamios nanofibrilares de PLA esterilizados por diferentes técnicas

ABSTRACT In recent years, tissue engineering has evolved considerably, due to the problems in the biomedical area concerning tissue regeneration therapies. Currently, work has been focused on the synthesis and physicochemical characterization of poly lactic acid scaffolds, a synthetic polyester that has been extensively study for its excellent biocompatibility and biodegradability. Moreover, sterilization strategies of scaffold are a crucial step for its application in tissue regeneration, however, the sterilization process have to maintain the structural and biochemical properties of the scaffold. Therefore, it is very important to carry out studies on the sterilization methods of the sample's material, since translational medicine is intended for in vivo applications. The aim of the present study was designed to analyze the effects of different sterilization techniques, i.e. ethylene oxide (ETO), gamma radiation (GR) and hydrogen peroxide- based plasma (H2O2) in biodegradable PLA scaffolds, and to determine the best sterilization technique to render a sterile product with minimal degradation and deformation, and good tissue response. Analysis of surface morphology showed that ETO and GR modified the PLA scaffolds without any change in its chemical composition. Moreover, the histological response showed that the scaffolds are biocompatible and those sterilized by GR showed a more severe inflammatory response, accompanied with the presence of giant foreign body cells. In conclusion, the results show that among sterilization techniques used in the preset study, the best results were observed with H2O2 sterilization, since it did not significantly modify the surface structure of the PLA fibers and their in vivo response did not cause an unfavorable tissue reaction.

RESUMEN En los últimos años, la ingeniería de tejidos ha evolucionado considerablemente, debido a las incógnitas en las terapias de regeneración en el área biomédica. Actualmente, se ha trabajado en la síntesis y caracterización fisicoquímica de andamios de poliácido láctico, el cual es un polímero sintético que se ha estudiado para aplicaciones en ingeniería de tejidos, debido a su biocompatibilidad y biodegradabilidad. El proceso de esterilización es un paso crucial en la aplicación de andamios en terapias de regeneración, sin embargo, la técnica de esterilización debe mantener las propiedades estructurales y bioquímicas del andamio. Por lo tanto, es muy importante realizar estudios sobre los métodos de esterilización de dichos andamios, ya que la medicina traslacional está diseñada para aplicaciones in vivo. El objetivo del presente estudio fue analizar los efectos de diferentes técnicas de esterilización como óxido de etileno (ETO), radiación gamma (GR) y plasma a base de peróxido de hidrógeno (H2O2) en andamios biodegradables de PLA, y determinar la mejor técnica de esterilización con mínima degradación y deformación, así como una respuesta tisular favorable. La estructura de la superficie de los andamios de PLA se modificó principalmente con las técnicas de óxido de etileno y radiación gamma, sin embargo, ninguna técnica modificó su composición química. Con la respuesta histológica se demostró que los andamios de PLA son biocompatibles y que los esterilizados por radiación gamma desencadenan una mayor respuesta inflamatoria y la formación de células gigantes de cuerpo extraño. En conclusión, los resultados muestran que las técnicas de esterilización utilizadas pueden modificar la morfología del andamio, sin embargo; los mejores resultados se observaron con la esterilización por plasma a base de peróxido de hidrógeno, ya que no modificó significativamente la estructura de la superficie de las fibras de PLA y su respuesta in vivo no provocó una reacción desfavorable en el tejido.

Does Sterilization Affect the Push Out Bond Strength of Experimental Dentin Posts? / ¿Afecta la esterilización la fuerza de adherencia de los postes de dentina experimentales?

ABSTRACT The purpose of this preliminary study is to evaluate the push-out bond strength (PBS) of experimental dentin posts (EDPs) obtained from human and bovine teeth sterilized by autoclaving and gamma radiation. Eighty-four mandibulary premolars were obturated and divided into three post groups: the glass fiber group (Fb) human EDP group (Hm) and the bovine EDP group (Bv). Three subgroups (n=12) were obtained for each EDP groups according to the sterilization methods; no sterilization (Cnt), steam autoclaving (Aut), and gamma radiation (Rad) a total dose of 25 kGy. All posts were cemented to root canals using a dual cured resin cement (Panavia SA). After the micro slices (1mm in thickness) were obtained of each subgroup, PBS test was performed. Data were analyzed using two-way ANOVA and Tukey's multiple comparison tests (α=.05). The post type and sterilization method was significantly effective on the PBS values according to the ANOVA (P<.001). No sterilization apllied EDP groups showed significantly higher PBS values than the sterilized groups (P<.001). Bv_Cnt (9.42 ±1.31) showed significantly lower PBS values than both Fb (12.36 ±1.54) and Hm_Cnt (11.06 ±1.38) groups (P<.001). Both steam autoclaving and gamma radiation affect the PBS values and fracture modes of EDPs negatively. The bovine EDPs are not as effective as human EDPs with regard to the PBS to the root dentin.

RESUMEN El propósito de este estudio preliminar es evaluar el push-out bond strength de los postes de dentina experimentales (PDE) obtenidos de dientes humanos y bovinos esterilizados por autoclave y radiación gamma. Ochenta y cuatro premolares mandibulares fueron obturadores y se dividieron en tres grupos experimentales de postes: el grupo de fibra de vidrio (Fb) grupo PDE humano (Hm) y el grupo PDE bovino (Bv). Se obtuvieron tres subgrupos (n=12) para cada grupo de PDE según los métodos de esterilización; sin esterilización (Cnt), autoclave a vapor (Aut) y radiación gamma (Rad) con una dosis total de 25 kGy. Todos los postes se cementaron a los conductos radiculares utilizando un cemento de resina de curado doble (Panavia SA). Después de que se obtuvieron las micro rebanadas (1 mm de espesor) de cada subgrupo, se realizó una prueba de push-out bond strength. Los datos se analizaron utilizando ANOVA de dos vías y las pruebas de comparación múltiple de Tukey (α=.05). El tipo de poste y el método de esterilización fueron significativamente efectivos en los valores de push-out bond strength según el ANOVA (P <.001). Los grupos de PDE no aplicados a la esterilización mostraron valores de push-out bond strength significativamente mayores que los grupos esterilizados (p <0,001). Bv_Cnt (9.42 ±1.31) mostró valores de push-out bond strength significativamente más bajos que los grupos Fb (12.36 ±1.54) y Hm_Cnt (11.06 ±1.38) (P <.001). Tanto el autoclave al vapor como la radiación gamma afectan negativamente los valores de push-out bond strength y los modos de fractura de los PDE. Los PDE bovinos no son tan efectivos como los PDE humanos con respecto a la fuerza de adhesión a la dentina de la raíz.

Development of a semi-empirical method to determine the efficiency of a gamma radiation detector for point sources / Desarrollo de un método semi-empírico para determinar la eficiencia de un detector de radiación gamma para fuentes puntuales

Abstract In the present work, we introduce the theoretical development and a new approach to validate a novel method to calibrate the efficiency of a gamma radiation detector for point sources, which we have named the "Efficiency Extrapolation Method". The method consists in the determination of the detector efficiency using a set of monoenergetic gamma sources, which we will refer to as reference efficiencies. From these values, we will extrapolate the detector efficiency to the complete energy range using the first principle physics of gamma radiation detection theory. Therefore the proposed method corresponds to a semi-empirical one. The determination of reference efficiencies must be done experimentally, but in this work, simulations were performed using FLUKA code. The reference energies will be: 59,54 keV, 661,66 keV and 1 274,54 keV associated to the isotopes 241Am, 137Cs and 22Na respectively. The second part of the method, the extrapolation from the reference energies to the gamma range, will be done over the energies emitted by 152Eu and 133Ba. In general, the results are very good. While the obtained results for energies 53,16 keV and greater than 344,3 keV show an excellent agreement, the results obtained for energies in the middle range are only good.

Resumen En el presente trabajo presentamos el desarrollo teórico y un nuevo enfoque para validar un nuevo método para calibrar la eficiencia de un detector de radiación gamma para fuentes puntuales, al cual hemos llamado el método de extrapolación de la eficiencia. El método consiste en la determinación de la eficiencia del detector usando un conjunto de fuentes gamma monoenergéticas, a las cuales llamaremos eficiencias de referencia. Desde estos valores, extrapolaremos la eficiencia del detector al rango completo de energías usando la física de primeros principios de la teoría de detección de radiación gamma. Por lo tanto el método propuesto corresponde a un método semiempírico. La determinación de las eficiencias de referencia debe hacerse experimentalmente, pero en este trabajo, se realizaron simulaciones usando el código FLUKA. Las energías de referencia serán: 59,54 keV, 661,65 keV y 1 274,54 keV asociadas a los isótopos 241 Am, 137Cs y 22Na respectivamente. La segunda parte del método, la extrapolación desde las energías de referencia hacia el rango gamma de las energías, será realizada sobre las energías emitidas por los isótopos 152Eu y 133Ba. En general, los resultados son muy buenos. Mientras que los resultados obtenidos para las energías de 53,16 keV y mayores que 344,3 keV muestran un excelente acuerdo, los resultados obtenidos para las energías del rango intermedio son solo buenas.

Mycotoxin residues in different chicken products by HPLC and their inactivation ‎using Gamma radiation

Mycotoxin residues were studied in different chicken products by HPLC and their inactivation using Gamma radiation. 120 differentchicken products were examined for the presence of mycotoxins (AFB1, AFB2, AFG1, AFG2, and OTA) using highly accurate and preciseHPLC-FLD assay. High recovery was achieved by purification of the sample extract using immunoaffinity columns (IAC). Derivatization ofAFs was carried out with a 0.005% aqueous solution of pyridine hydrobromide perbromide (PBPB) by utilizing a post-column LC pump.The incidence of mycotoxins in analyzed chicken products showed that the level of Afla B1, Afla B2, and Afla G2 were 10 % for each and ofAfla G1 and Ochra A were ranged from 10 to 16.7% for each. The results showed that the highest concentration of examined mycotoxinswas present in the liver as the liver is the harbor site of mycotoxin residues. The influence of different gamma irradiation rays (6, 8, and 10kGy) was studied on the reduction of the existed in examined chicken products. There is a positive connection between the elevation ofgamma irradiation dose used to the samples and the reduction level of total mycotoxins current in these samples, whereby, the mostreduction percentage of mycotoxins were achieved at 10 kGy; it reaches 19.6% for total mycotoxins, 27% for AF B1, 40.43% for AF B2,59.42% for AF G1, 92.15% for AF G2, and 73.44% for OTA.

Gene coexpression analysis reveals dose-dependent and type-specific networks responding to ionizing radiation in the aquatic model plant Lemna minor using public data

Ionizing radiations (IRs) are widespread damaging stresses to plant growth and development. However, the regulatory networks underlying the mechanisms of responses to IRs remains poorly understood. Here, a set of publicly available transcriptomic data (conducted by Van Hoeck et al. 2015a), in which Lemna minor plants were exposed to a series of doses of gamma, beta and uranium treatments was used to perform gene coexpression network analysis. Overall, the genes involved in DNA synthesis and chromatin structure, light signalling, photosynthesis, and carbohydrate metabolism were commonly responsive to gamma, beta and uranium treatments. Genes related to anthocyanin accumulation and trichome differentiation were specifically downregulated, andgenes related to nitrogen and phosphate nutrition, cell vesicle transport, mitochondrial electron transport and ATP synthesis were specifically upregulated in response to uranium treatment. While genes involved in DNA damage and repair, RNA processing and RNA binding were specifically downregulated and genes involved in calcium signalling, redox and degradation of carbohydrate metabolism were specifically upregulated responding to gamma radiation. These findings revealed both dose-dependent and typespecific networks responding to different IRs in L. minor, and can be served as a useful resource to better understand the mechanisms of responses to different IRs in other plants.

Heartwood and sapwood in eucalyptus trees: non-conventional approach to wood quality

ABSTRACT This study evaluated the quality of heartwood and sapwood from mature trees of three species of Eucalyptus, by means of the qualification of their proportion, determination of basic and apparent density using non-destructive attenuation of gamma radiation technique and calculation of the density uniformity index. Six trees of each species (Eucalyptus grandis - 18 years old, Eucalyptus tereticornis - 35 years old and Corymbia citriodora - 28 years old) were used in the experimental program. The heartwood and sapwood were delimited by macroscopic analysis and the calculation of areas and percentage of heartwood and sapwood were performed using digital image. The uniformity index was calculated following methodology which numerically quantifies the dispersion of punctual density values of the wood around the mean density along the radius. The percentage of the heartwood was higher than the sapwood in all species studied. The density results showed no statistical difference between heartwood and sapwood. Differently from the density results, in all species studied there was statistical differences between uniformity indexes for heartwood and sapwood regions, making justifiable the inclusion of the density uniformity index as a quality parameter for Eucalyptus wood.

Gamma radiation treatment activates glucomoringin synthesis in Moringa oleifera

Abstract Plants are a very rich source of pharmacologically relevant metabolites. However, the relative concentrations of these compounds are subject to the genetic make-up, the physiological state of the plant as well as environmental effects. Recently, metabolic perturbations through the use of abiotic stressors have proven to be a valuable strategy for increasing the levels of these compounds. Oxidative stress-associated stressors, including ionizing radiation, have also been reported to induce metabolites with various biological activities in plants. Hence, the aim of the current study was to investigate the effect of gamma radiation on the induction of purported anti-cancerous metabolites, glucomoringin and its derivatives, in Moringa oleifera Lam., Moringaceae. Here, an UHPLC-qTOF-MS-based targeted metabolic fingerprinting approach was used to evaluate the effect of gamma radiation treatment on the afore-mentioned health-beneficial secondary metabolites of M. oleifera. Following radiation, an increase in glucomoringin and three acylated derivatives was noted. As such, these molecules can be regarded as components of the inducible defense mechanism of M. oleifera as opposed to being constitutive components as it has previously been assumed. This might be an indication of a possible, yet unexplored role of moringin against the effects of oxidative stress in M. oleifera plants. The results also suggest that plants undergoing photo-oxidative stress could accumulate higher amounts of glucomoringin and related molecules.

Alarma gamma para la protección radiológica / A Gamma Alarm system for radiation protection

En el presente trabajo se describe el desarrollo de un instrumento para el monitoreo radiológico identificado como Alarma Gamma (GAMAL01). Estos monitores detectan el incremento de la tasa de conteo de la radiación y emiten una alarma cuando esta magnitud sobrepasa un umbral determinado. El umbral para la emisión de la alarma se fija de manera que esta se produzca cuando se considere que la tasa de conteo de la radiación pueda provocar una exposición inaceptable del personal a las radiaciones, de acuerdo a lo establecido por las normas de seguridad radiológica vigentes en el país. La señal de alarma indica la necesidad de realizar la evacuación del personal del sitio afectado. El instrumento consta de dos secciones: una analógica y otra digital

This paper describes the development of an instrument for radiological monitoring, identified as Gamma Alarm (GAMAL01). The instrument monitors the increment of radiation count rate and produces an alarm signal, when the count rate becomes superior to a fixed threshold value. The threshold is fixed to assure that the signal emission will start when the value of the radiation count rate may cause a non acceptable radiological exposition of the personnel, according to the radiological safety regulations existing in the country. Following the signal the personnel must be evacuated from the affected place. The instrument consists of two sections, an analogical section and a digital one

Efeitos da irradiação gama sobre a estabilidade físico-química de anéis de lula congelados / Effects of gamma irradiation on the physico-chemical stability of frozen squid rings

O principal objetivo deste trabalho foi avaliar os efeitos do processo de exposição à radiação gama (césio- 137) na estabilidade físico-química de anéis de lula, Dorytheutis plei, congelados. Foram investigadas 30 amostras, separadas em três grupos, de acordo com a dose de radiação absorvida: 0 kGy (controle), 1,5 kGy e 3,0 kGy. Análises físico-químicas, de Base Voláteis Totais e pH, relevantes na determinação da qualidade do pescado, foram realizadas durante o período de seis semanas. Durante a estocagem, observou-se uma relativa estabilidade físico-química nas amostras estudadas, caracterizando um produto viável para a comercialização. Dentre as doses investigadas, 1,5 kGy mostrou-se a mais apropriada para tratamento do produto. Não houve alteração significativa da composição centesimal das amostras.

Síntesis y toxicidad de hidrogeles de poli(metacrilato de 2-hidroxietilo-co-acrilamida) obtenidos mediante irradiación con cuantos gamma / Sinthesys and toxicity of poly (2-hydroxyethyl methacrylate-co-acrylamide) hydrogels obtained by gamma photon irradiation

En el presente trabajo se reporta la obtención de hidrogeles de copolímeros de polimetacrilato de 2-hidroxietilo-co-acrilamida mediante la copolimerización y reticulación simultánea a partir de sus correspondientes monómeros y mediante irradiación con cuantos gamma. La composición del copolímero se determinó mediante análisis elemental y espectroscopia infrarroja. Adicionalmente se muestran los resultados de toxicidad de sus matrices, para ello se evaluó la acrilamida libre y se realizó el ensayo SOS en bacterias. Se comprobó que los extractos de lavados de las matrices de estos copolímeros contenían muy bajos niveles de acrilamida libre y no mostraron efecto tóxico en las células de E. coli para ninguno de los tiempos evaluados

In this paper is reported the obtaining of hydrogels from poly hydroxyethyl methacrylate-co-acryla-mide, by simultaneous crosslinking copolymerization from corresponding monomers by gamma photon irradiation. The composition of network chains in the copolymer was estimated from elemental analysis and infrared spectroscopy. Additionally, the toxicity of these materials was tested finding the free acrylamide and biological assay SOS in bacteria. The extracts of washed matrices of copolymers showed low level of free acrylamide. Moreover, they were non toxic on the E. coli cells tested at any time

Black tea extract protects against -radiation-induced membrane damage of human erythrocytes.

The membrane integrity of circulating red blood cells (RBCs) is compromised by the deleterious actions of -radiation in humans. Tea is the most widely consumed popular, inexpensive and non-toxic beverage rich in antioxidants. Here, we explored the radioprotective actions of black tea against the -radiation-induced membrane permeability of human erythrocytes. The phytochemical analysis of tea revealed the total polyphenol content to be 114.89±6.03 mg gallic acid equivalent/g dry wt. and flavonoid content, 34±0.11 mg catechin equivalent/g dry wt. of the extractable solid in the commercially available tea bags. Tea extracts showed potential scavenging of H2O2 and NO, appreciable extent of total antioxidant capacity and effective anti-hemolytic action. Tea extracts (15 µg/mL) significantly ameliorated the -radiation-induced increase of the levels of thiobarbituric acid-reactive substances (TBARS, an index of lipid peroxidation) in the RBC membrane ghosts. Stored blood showed higher levels of K+ ion as compared to the normal blood which was elevated by -radiation. Membrane ATPase was inhibited by the exposure to -radiation. Treatment of RBCs with the tea extracts (15 µg/ml) prior to the exposure of -radiation significantly mitigated these changes in the erythrocyte membranes caused by the lower dose of radiation (4 Gy) as compared to that induced by the higher dose of -radiation.

Effect of Gamma Radiation on Growth and Metabolic Activities of Arthrospira platensis

This work aimed to study the influence of gamma radiation on the growth and production of some active substances of Arthrospira platensis. Biomass production was significantly inhibited (p ≤ 0.05) by 21 and 34%, with respect to the control at 2.0 and 2.5 kGy, respectively. Chlorophyll-a content showed 11% reduction at 2.5 kGy compared to the control. As a result of growth and Chl-a inhibition, chlorophyll productivity recorded a continuous significant decrease below the control in the cells exposed to 1, 1.5, 2 and 2.5 kGy by 8, 12, 15 and 25%, respectively after 15 days of incubation. In addition, phycobillins productivity showed significant decrease by 10 and 36% below the control at 2 and 2.5 kGy of gamma radiation, respectively. Protein production decreased significantly by 24% at 1.5 kGy; low doses of gamma irradiation (0.5, 1.0 and 1.5 kGy) induced carbohydrate production by 106, 246 and 146%, respectively. Lipid content increased significantly over the control at 0.5 kGy of gamma irradiation by 22%, which was decreased at higher doses. Interestingly, carotenoid productivity showed significant increase at all used gamma doses up to 155% over the control.

New insights into the structural characteristics of irradiated crotamine

Background:Since ionizing radiation has the potential to alter the molecular structure and affect the biologica properties of biomolecules, it has been successfully employed to attenuate animal toxins. The present study aimed to characterize the structural modifications on irradiated crotamine, a toxin from Crotalus durissus terrificus venom, using circular dichroism (CD), fluorescence, Fourier transformed infrared spectroscopy (FTIR), atomic force microscopy (AFM) and differential scanning calorimetry (DSC).Methods:A combination of size exclusion and ion-exchange chromatography was used to purify the peptide using crude venom. The pure toxin was then submitted to 2 kGy gamma irradiation doses from a cobalt-60 source. Native and irradiated crotamine were analyzed using a fluorescence spectrophotometer. Wavelength was fixed at 295 nm and fluorescence emission scans were collected from 300 to 400 nm. CD and FTIR techniques were used to identify the secondary structure of both samples. DSC analyses were performed at a starting temperature of 20 °C up to a final temperature of 90 °C. AFM provided a 3D profile of the surfaces of both crotamine forms adsorbed on mica.Results:Fluorescence spectroscopy showed that the quantum yield of the irradiated form decreased. CD spectra of native and irradiated crotamine solutions showed differences between the samples in wavelength, indicating that irradiation induced a transition of a small portion of the random coil regions towards an a-helical conformation. FTIR and CD showed that the native and irradiated crotamine spectra were different with regard to secondary structure. The thermodynamic analysis showed that irradiation caused changes in the calorimetric profile and CD showed that temperature-induced changes also occur in the secondary structure. Finally, AFM showed the possible formation of insoluble aggregates.Conclusions:Our results indicate that irradiation leads to progressive changes in the structure of the toxin, which could explain a decrease in myotoxic activity.(AU)

New insights into the structural characteristics of irradiated crotamine

Background: Since ionizing radiation has the potential to alter the molecular structure and affect the biologica properties of biomolecules, it has been successfully employed to attenuate animal toxins. The present study aimed to characterize the structural modifications on irradiated crotamine, a toxin from Crotalus durissus terrificus venom, using circular dichroism (CD), fluorescence, Fourier transformed infrared spectroscopy (FTIR), atomic force microscopy (AFM) and differential scanning calorimetry (DSC). Methods: A combination of size exclusion and ion-exchange chromatography was used to purify the peptide using crude venom. The pure toxin was then submitted to 2 kGy gamma irradiation doses from a cobalt-60 source. Native and irradiated crotamine were analyzed using a fluorescence spectrophotometer. Wavelength was fixed at 295 nm and fluorescence emission scans were collected from 300 to 400 nm. CD and FTIR techniques were used to identify the secondary structure of both samples. DSC analyses were performed at a starting temperature of 20 °C up to a final temperature of 90 °C. AFM provided a 3D profile of the surfaces of both crotamine forms adsorbed on mica. Results: Fluorescence spectroscopy showed that the quantum yield of the irradiated form decreased. CD spectra of native and irradiated crotamine solutions showed differences between the samples in wavelength, indicating that irradiation induced a transition of a small portion of the random coil regions towards an a-helical conformation. FTIR and CD showed that the native and irradiated crotamine spectra were different with regard to secondary structure. The thermodynamic analysis showed that irradiation caused changes in the calorimetric profile and CD showed that temperature-induced changes also occur in the secondary structure. Finally, AFM showed the possible formation of insoluble aggregates. Conclusions: Our results indicate that irradiation leads to progressive changes in the structure of the toxin, which could explain a decrease in myotoxic activity.

Comparative Occurrence of Resident Fungi on Gamma Irradiated and Steam Sterilized Sorghum Grains (Sorghum bicolor L.) for Spawn Production in Ghana.

Sorghum is one of the important cereals consumed by humans, animals and also used for the production of mushroom spawns in Ghana. Aim: Identification of fungi present on sorghum grains before and after pretreatment (steam and gamma radiation) principally for mushroom cultivation Methodology: The total number of mycoflora (Log10 CFU g-1) of sorghum grains and their relative frequency (percentage occurrence) associated with the raw grains and the mycoflora present after subjecting the sorghum grains to gamma radiation doses of 0, 5, 10, 15, 20, 25 and 32 kGy at a dose rate of 1.7 kGy/h from a Cobalt-60 source (SLL-515, Hungary) and moist heat at a temperature of 100- 120ºC for 2- 2.5 hours was evaluated. Mycological analysis was done by direct plating method on Cooke’s and Dichloran Rose Bengal Chloramphenicol (DRBC) media. Results: Nine fungal species belonging to six genera were associated with the sorghum grains. Among these fungi were Cladosporium macrocarpum, Trichoderma harzianum, Fusarium oxysporum, Rhodotorula spp., Penicillium spp., Aspergillus niger, Aspergillus fumigatus, Aspergillus ochraceous and Aspergillus flavus. Comparatively higher fungal counts of 3.27 and 3.82 Log10 CFU g-1 were recorded for non-pretreated while lower counts of 0.5 Log10 CFU g-1 were recorded for pretreated sorghum grains. Gamma radiation and moist heat significantly (P<0.05) reduced total fungal populations by an average of 2.4 and 2.1 log cycles, respectively. Rhodotorula sp. (11.5%), Penicillium sp. (34.6%), Aspergillus fumigatus (29.9%) persisted on the moist heat sterilized while only Rhodotorula sp. (100%) persisted on gamma irradiated grains. Conclusion: These data indicate possible health hazards for humans and animals upon consumption of such contaminated food grain by toxigenic moulds and also reveal the sensitivity of fungal species to gamma radiation and moist heat as a selective substrate for oyster mushroom spawn preparation.

Avaliação biológica de machos esterilizados por radiação gama, para utilização em programas de controle populacional de Aedes aegypti (DIPTERA: CULICIDAE) / Biological evaluation of sterilized males by gamma radiation for use in the Aedes aegypti (Diptera: Culicidae) population control programs

Este estudo avaliou o desempenho biológico e reprodutivo de machos de Aedes aegypti esterilizados por radiação gama e seu potencial para uso no controle da espécie, através da técnica do inseto estéril (TIE). A eficácia de esterilização de diferentes doses de radiação gama, 30, 40 e 50 Gy, foi avaliada em machos irradiados (MI) na fase de pupa. O seu efeito sobre a longevidade e competitividade de acasalamento dos MI, sobre a fecundidade e fertilidade das fêmeas acasaladas com estes machos, bem como de fêmeas também irradiadas (FI), foram parâmetros analisados. Os testes de competitividade foram conduzidos em gaiolas teladas (2 X 2 m) em condições simuladas de campo, onde foram liberados machos irradiados e machos não irradiados (MNI), nas seguintes proporções: 5:1, 10:1 e 15:1. Os resultados revelaram que o fitness biológico e reprodutivo dos mosquitos são afetados pela radiação, de modo que os efeitos são diretamente proporcionais a dose de radiação utilizada. O parâmetro de longevidade apresentou diferenças significativas, tanto quando os MI foram analisados de forma individual quanto em grupo. Para a maior dose, MI e FI viveram em média 19,6 ± 2,2 dias e 25,8 ± 2,2 dias, respectivamente, enquanto que no grupo controle, estes valores foram de 24,9 ± 2,5 dias e 30,4 ± 2,5 dias...

This study evaluated the biological and reproductiv e performance of Aedes aegyptimales sterilized by gamma radiation and its potential for use in controlling the species through the Sterile Insect Technique (SIT). The sterilization efficacy of different doses of gamma radiation, 30, 40 and 50 Gy, was assessed in irradiated males (IM) in the pupal stage. The direct effects of gama radiation on the longevity and competitiveness of IM, as well as the effect on the fecundity and fertility of females mated with these males, and also irradiated females (IF), were analyzed. The competitiveness was examined by tests conducted in large cage (2 x 2 meters) in semi field conditions where irradiated males (IM) and non-irradiated males (NIM) were released in the following proportions: 5:1, 10:1 and 15:1. The results revealed that the biological and reproductive fitness of the mosquitoes are affected by radiation, so that the effects are directly proportional to the radiation dose. The longevity parameter showed significant differences...

O papel de RhoA e Rac1 GTPases nas respostas celulares após danos no DNA induzidos por radiação ionizante gama / The role of RhoA and Rac1 GTPases in cellular responses after DNA damage induced by ionizing gamma radiation

O mecanismo pelo qual uma célula responde a algum dano no seu material genético é extremamente importante. Isto ocorre pela rápida ativação da maquinaria de reparo de danos no DNA, a qual é composta por uma rede intrincada de sinalização proteica, culminando no reparo do DNA; porém se o dano for irreparável ocorre ativação de mecanismos de morte celular. RhoA,e Rac1 pertencem a família das pequenas proteínas sinalizadoras Rho GTPases, as quais atuam como interruptores moleculares ciclando entre estado ativo (ligada a GTP) e inativo (ligada a GDP). Os componentes desta família estão relacionados ao controle dos mais diversos processos celulares como, por exemplo, remodelamento do citoesqueleto, migração, adesão, endocitose, progressão do ciclo celular e oncogênese. No entanto, apesar das proteínas Rho GTPases estarem envolvidas em um amplo espectro de atividades biológicas, há poucas informações sobre seu papel na manutenção da integridade genômica quando células são submetidas a algum agente genotóxico. Para investigar o envolvimento das GTPases RhoA e Rac1 nas respostas de células submetidas a radiação gama, foram gerados, a partir de células de carcinoma de cervix humano - HeLa, sublinhagens clonais mutantes de RhoA e Rac1 expressando exogenamente RhoA constitutivamente ativa (HeLa-RhoA V14), RhoA dominante negativa (HeLa-RhoA N19), Rac1 constitutivamente ativa (HeLa-Rac1 V12) e Rac1 dominante negativa (HeLa-Rac N17). Após estas linhagens celulares serem expostas a diferentes doses de radiação gama, observamos que ambas GTPases, RhoA e Rac1, são ativadas em resposta aos efeitos da radiação. Além disso, a modulação da atividade destas enzimas, através das mutações, levou a uma alteração das respostas celulares frente aos danos no DNA, como uma redução da capacidade de reparar quebras simples e duplas nas fitas do DNA. Por outro lado, a deficiência de RhoA ou Rac1 GTPase levou a uma redução da ativação de Chk1 e Chk2 ou da fosforilação da histona H2AX, respectivamente, prejudicando os mecanismos de detecção de danos no DNA e levando as células a permanecerem mais tempo nos pontos de checagem G1/S e/ou G2/M do ciclo celular. Esses fatores contribuíram de modo expressivo para a redução da proliferação e sobrevivência celular levando as células à morte. Por fim, ensaios celulares de reparo de danos de um DNA exógeno através de mecanismos de Recombinação Homóloga (HR) e Recombinação Não-Homóloga de extremidades (NHEJ), demonstraram que a inibição da atividade de RhoA reduz significativamente a eficiência de ambas vias de reparo. Desta maneira, este trabalho demonstra e reforça a existência de mais um viés de atuação das pequenas GTPases RhoA e Rac1, agora em células HeLa, nas respostas celulares aos danos induzidos por exposição a radiação gama, modulando a sobrevivência, proliferação e indiretamente modulando resposta ao reparo do DNA através da via de Recombinação Homóloga e Não-Homóloga

The mechanism by which a cell responds to DNA damage is extremely important. This occurs by a quick activation of the DNA damage repair machinery, which consists of an intricate protein signaling network culminating in DNA repair. But if the damages are irreparable occurs there is activation of cell death mechanisms. RhoA and Rac1 belong to family of small Rho GTPases, signaling proteins that act as molecular switches cycling between the active state (GTP-bound) and inactive state (GDP-bound). Members of this family are implicated in the control of diverse cellular process such as cytoskeletal remodeling, migration, adhesion, endocytosis, cell cycle progression, and oncogenesis. However, despite Rho proteins are involved in a broad spectrum of biological activities, there is just a few information about their roles in the maintenance of genomic integrity, that is, when the cells are subjected to some kinf of genotoxic agent. To investigate the involvement of the GTPases RhoA and Rac1 in cellular responses to gamma radiation, we generated from human cervix carcinoma cells - HeLa, clonal sublines of RhoA and Rac1 mutants, exogenous and stably expressing the constitutively active RhoA (HeLa-RhoA V14), the dominant negative RhoA (HeLa-RhoA N19), the constitutively active Rac1 (HeLa-Rac1 V12) and the dominant negative Rac1 (HeLa-Rac1 N17). After all these cell lines have been exposed to different doses of gamma radiation, we found that both GTPases, RhoA and Rac1, are activated in response to the radiation effects. Furthermore, the modulation of two enzymes activity, by using the mutant clones, led to a change in cellular responses to the DNA damage, as the reduction in the capacity of repairing DNA single and double strand breaksr. On the other hand, the deficiency of RhoA or Rac1 GTPase led to a reduction of Chk1 and Chk2 activation, or on the phosphorylation of histone H2AX, respectively, hindering the mechanisms of DNA damage detection and arresting cells in the G1/S and/or G2/M checkpoints of cell cycle. These factors significantly contributed to the reduction of cell proliferation and survival, leading cells to death. Finally, cellular assays of DNA damage repair of exogenous DNA by Homologous Recombination (HR) and Non-Homologous End Joining (NHEJ), demonstrated that RhoA inhibition significantly reduced the repair efficiency of both pathways. Thus, this work demonstrates and reinforces the existence of other biological functions of small GTPases RhoA and Rac1 in HeLa cells, by regulating cellular responses to DNA damage induced by exposure to gamma radiation, modulating the survival, proliferation and indirectly modulating the response to DNA damage repair pathway through the Homologous Recombination and Non-Homologous Recombination

Application of radiation technology in vaccines development

One of the earliest methods used in the manufacture of stable and safe vaccines is the use of chemical and physical treatments to produce inactivated forms of pathogens. Although these types of vaccines have been successful in eliciting specific humoral immune responses to pathogen-associated immunogens, there is a large demand for the development of fast, safe, and effective vaccine manufacturing strategies. Radiation sterilization has been used to develop a variety of vaccine types, because it can eradicate chemical contaminants and penetrate pathogens to destroy nucleic acids without damaging the pathogen surface antigens. Nevertheless, irradiated vaccines have not widely been used at an industrial level because of difficulties obtaining the necessary equipment. Recent successful clinical trials of irradiated vaccines against pathogens and tumors have led to a reevaluation of radiation technology as an alternative method to produce vaccines. In the present article, we review the challenges associated with creating irradiated vaccines and discuss potential strategies for developing vaccines using radiation technology.