RESUMEN
This paper reports a lab-on-a-chip for the detection of Sarin nerve agent based on rapid electrochemical detection. The chemical warfare agent Sarin (C4H10FO2P, O-isopropyl methylphosphonofluoridate) is a highly toxic organophosphate that induces rapid respiratory depression, seizures and death within minutes of inhalation. As purified Sarin is colourless, odourless, water soluble and a easily disseminated nerve agent, it has been used as a weapon in terrorist or military attacks. To ascertain whether potable water supplies have been adulterated with this extremely potent poison, an inexpensive, sensitive and easy to use portable test kit would be of interest to first responders investigating such attacks. We report here an amperometric-based approach for detecting trace amounts of Sarin in water samples using a screen-printed electrode (SPE) integrated in a microfluidic chip. Enzymatic inhibition was obtained by exposing the immobilised biosensor in the microfluidic platform to Sarin in water samples. With the aid of cobalt phthalocyanine modified SPE, the device could detect Sarin at part-per-billion levels with concentration as low as 1 nM. The detection method reported here represents a significant improvement over the authors'previous optical-based detection method.
Asunto(s)
Sustancias para la Guerra Química/análisis , Técnicas Electroquímicas , Indoles/química , Dispositivos Laboratorio en un Chip , Sarín/análisis , Técnicas Electroquímicas/instrumentación , Técnicas Electroquímicas/métodos , Isoindoles , Sensibilidad y EspecificidadRESUMEN
Ricin is a toxin that can be easily extracted from seeds of Ricinus communis plants. Ricin is considered to be a major bio-threat as it can be freely and easily acquired in large quantities. A deliberate release of such toxin in civilian populations would very likely overwhelm existing public health systems, resulting in public fear and social unrest. There is currently no commercially available or FDA-approved prophylaxis such as vaccines, or therapeutic antitoxins or antidotes, available for ricin intoxication. Patient treatment is typically supportive care based on symptoms, often designed to reinforce the body's natural response. This paper describes the development and validation of a robust ELISA test kit, which can be used to screen for ricin in biological specimens such as whole blood and faeces. Faecal specimens are shown in this study to have better diagnostic sensitivity and a wider diagnostic window compared to whole blood. From these results, it is concluded that faeces is the most suitable clinical specimen for diagnosis of ricin poisoning via the oral route. The ELISA test kit can also detect ricin in environmental samples. An advantage of this ELISA kit over other commercial off-the-shelf (COTS) detection kits currently on the market that are developed to screen environmental samples only is its ability to diagnose ricin poisoning from clinical specimens as well as detect ricin from environmental samples.
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Anticuerpos Monoclonales/química , Armas Biológicas , Sustancias para la Guerra Química/aislamiento & purificación , Ensayo de Inmunoadsorción Enzimática/normas , Juego de Reactivos para Diagnóstico/normas , Ricina/sangre , Animales , Heces/química , Humanos , Límite de Detección , Masculino , Variaciones Dependientes del Observador , Conejos , Ratas , Ratas Wistar , Reproducibilidad de los ResultadosRESUMEN
BACKGROUND: Early in the COVID-19 pandemic, there was a global shortage of masks. Although mask reprocessing was practiced, no clinical study has assessed systematically the impact of repeated cycles of wear and decontamination on the integrity of N95 filtering facepiece respirators (FFRs). METHODS: We evaluated mask fit assessed by qualitative respirator fit test (QRFT) after each cycle of wear and decontamination, as well as four measures of mask integrity-bacterial filtration efficacy, particle filtration efficacy, differential pressure, and splash resistance through five cycles of wear and decontamination using one of the four modalities (moist heat, steam, ultraviolet-C irradiation, and hydrogen peroxide plasma). RESULTS: A total of 60.6% (hydrogen peroxide plasma) to 77.5% (moist heat) of the FFRs passed five cycles of wear and decontamination, as assessed by the wearers passing QRFT all five times. Moist heat-decontaminated FFRs retained all technical measures of integrity through all five cycles. CONCLUSIONS: This is the first large-scale study to assess systematically the impact (clinically and quantitatively) on N95 FFR integrity of repeated cycles of wearing followed by decontamination. Our results suggest that moist heat is a promising method for decontaminating N95 FFRs. Performing QRFT after every cycle of wear and decontamination ensures wearer safety. Although there is currently no mask shortage, reprocessing may reduce medical waste and improve sustainability.
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Respiradores N95 , Dispositivos de Protección Respiratoria , Humanos , Vapor , Peróxido de Hidrógeno , Calor , Descontaminación/métodos , Pandemias/prevención & control , Equipo Reutilizado , MáscarasRESUMEN
Paper-based enzyme immobilization for a flow injection electrochemical biosensor integrated with a reagent-loaded cartridge toward a portable device was developed. A paper disk was immobilized with enzyme, then it was integrated in a flow cell as an electrochemical biosensor. A silicon tube reagent-loaded cartridge was integrated into the system, a complicated procedure was simplified as a one-click operation toward development for point-of-care applications. In this research, glucose oxidase (GOx) was employed as a model enzyme, silver ion as an inhibition reagent for GOx, and EDTA as a regeneration reagent. When GOx was inhibited by silver ions, glucose was introduced for electrochemical measurements before and after inhibited enzyme regeneration and the difference was caused by silver inhibition. The modular device has great potential for other applications, e.g., detection of enzyme activity and substrate. The platform based on double-test mode provided accurate results due to elimination of an average or control value in comparison with classical routine approaches.
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Técnicas Biosensibles/instrumentación , Electroquímica/instrumentación , Enzimas Inmovilizadas/química , Enzimas Inmovilizadas/metabolismo , Análisis de Inyección de Flujo/instrumentación , Papel , Integración de Sistemas , Aspergillus niger/enzimología , Biocatálisis , Inhibidores Enzimáticos/farmacología , Enzimas Inmovilizadas/antagonistas & inhibidores , Diseño de Equipo , Glucosa Oxidasa/antagonistas & inhibidores , Glucosa Oxidasa/química , Glucosa Oxidasa/metabolismo , Sistemas de Atención de Punto , Plata/análisis , Plata/químicaRESUMEN
Victims exposed to sulfur mustard (HD) in World War I and Iran-Iraq war, and those suffered occupational or accidental exposure have endured discomfort in the respiratory system at early stages after exposure, and marked general physical deterioration at late stages due to pulmonary fibrosis, bronchiolitis obliterans or lung cancer. At molecule levels, significant changes of cytokines and chemokines in bronchoalveolar lavage and serum, and of selectins (in particular sE-selectin) and soluble Fas ligand in the serum have been reported in recent studies of patients exposed to HD in Iran-Iraq war, suggesting that these molecules may be associated with the pathophysiological development of pulmonary diseases. Experimental studies in rodents have revealed that reactive oxygen and nitrogen species, their product peroxynitrite (ONOO(-)), nitric oxide synthase, glutathione, poly (adenosine diphosphate-ribose) polymerase, activating protein-1 signaling pathway are promising drug targets for preventing HD-induced toxicity, whereas N-acetyl cysteine, tocopherols, melatonin, aprotinin and many other molecules have been proved to be effective in prevention of HD-induced damage to the respiratory system in different animal models. In this paper, we will systemically review clinical and pathophysiological changes of respiratory system in victims exposed to HD in the last century, update clinicians and researchers on the mechanism of HD-induced acute and chronic lung damages, and on the relevant drug targets for future development of antidotes for HD. Further research directions will also be proposed.
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Bronquiolitis Obliterante/fisiopatología , Sustancias para la Guerra Química/toxicidad , Neoplasias Pulmonares/fisiopatología , Gas Mostaza/toxicidad , Fibrosis Pulmonar/fisiopatología , Sistema Respiratorio/efectos de los fármacos , Animales , Bronquiolitis Obliterante/etiología , Modelos Animales de Enfermedad , Selectina E/metabolismo , Glutatión/metabolismo , Humanos , Irán , Irak , Neoplasias Pulmonares/etiología , Óxido Nítrico Sintasa/metabolismo , Ácido Peroxinitroso/metabolismo , Poli(ADP-Ribosa) Polimerasas/metabolismo , Fibrosis Pulmonar/etiología , Primera Guerra MundialRESUMEN
The present study investigated the protective actions of des-aspartate-angiotensin I (DAA-I) in mice that were intranasally administered 2-chloroethyl ethyl sulfide (CEES), a half sulfur mustard. The protection was dose-dependent, and an oral dose of 75 mg kg⻹ per day administered 18 h post exposure and for the following 13 days, offered maximum protection that increased survival by a third. DAA-I attenuated the early processes of inflammation seen in the CEES-inoculated mice. DAA-I attenuated (i) elevated pulmonary ROS, and gp91-phox protein of NADPH oxidase, a non phagocytic enzyme that generates superoxide and subsequent ROS; (ii) intercellular adhesion molecule-1 (ICAM⻹) that is involved in the extravasation of circulating leucocytes; and (iii) myeloperoxidase activity, which is a surrogate enzymatic measurement of neutrophil infiltration. These actions led to improved histological lung structures, and survival of type-1 pneumocytes. The action of DAA-I on animal survival was blocked by losartan, a selective angiotensin AT1 receptor blocker, indicting that the AT1 receptor mediates the protection. The presence of elevated PGE2 and PGI2 in lung supernatants of DAA-I treated CEES-inoculated mice indicates that the two prostaglandins are involved in signaling the protective actions of DAA-I. This finding complements earlier studies showing that DAA-I acts on an indomethacin-sensitive angiotensin AT1 receptor. The findings of the present study are the first demonstration of an angiotensin peptide as an effective antidote for CEES intoxication. DAA-I is also an effective therapeutic intervention against CEES that was instituted at 18 h post exposure, and challenges conventional assumptions of limited efficacy with delayed action against alkylating agents.
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Angiotensina I/análogos & derivados , Pulmón/efectos de los fármacos , Pulmón/patología , Gas Mostaza/análogos & derivados , Angiotensina I/farmacología , Animales , Líquido del Lavado Bronquioalveolar/química , Dinoprostona/metabolismo , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Epoprostenol/metabolismo , Molécula 1 de Adhesión Intercelular/metabolismo , Losartán/farmacología , Masculino , Ratones , Ratones Endogámicos BALB C , Gas Mostaza/toxicidad , NADPH Oxidasas/metabolismo , Peroxidasa/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Receptores Inmunológicos/metabolismo , Transducción de SeñalRESUMEN
OBJECTIVES: We have characterised the population pharmacokinetics-pharmacodynamics of pyridostigmine given as pyridostigmine bromide. METHODS: Over three days 50 healthy Chinese male subjects each received seven doses of 30 mg pyridostigmine bromide orally (3 x 10 mg every 8 h). Plasma concentrations of pyridostigmine and red blood cell acetylcholinesterase (AChE) activity were determined at various times within the eight hours after the first and the seventh doses. The resulting pharmacokinetic data were fitted to a single compartment open model with first-order absorption and elimination. The pharmacodynamics were modelled using an inhibitory E(max) model. The potential influence of demographic and biological covariates on the model parameters was investigated. Nonlinear mixed effects modelling was performed using NONMEM. KEY FINDINGS: The apparent clearance and volume of distribution as well as absorption rate constant of plasma pyridostigmine were estimated to be 136 l/h, 130 l and 0.68 1/h, respectively. The maximum red blood cell AChE activity decrease (E(max)) and plasma pyridostigmine concentration producing 50% of this reduction (EC50) were estimated to be 9.32 AChE units per gram haemoglobin and 51.9 ng/ml, respectively. None of the tested covariates were found to be correlated with any of the model parameters. Dosing simulations suggested that 30 mg repeated every six hours might be needed to achieve steady-state trough percentage inhibition above the recommended 10% in healthy Chinese males. CONCLUSIONS: The pharmacokinetics and the effects of pyridostigmine on red blood cell AChE activity were described using a mixed effects model. For Chinese males, the dosing interval may have been shorter than that recommended for the Caucasian population. Additional studies are needed to confirm these findings.
Asunto(s)
Acetilcolinesterasa/efectos de los fármacos , Inhibidores de la Colinesterasa/farmacología , Inhibidores de la Colinesterasa/farmacocinética , Bromuro de Piridostigmina/farmacología , Bromuro de Piridostigmina/farmacocinética , Acetilcolinesterasa/sangre , Adulto , Pueblo Asiatico , Cálculo de Dosificación de Drogas , Humanos , Modelos Lineales , Masculino , Dinámicas no Lineales , Bromuro de Piridostigmina/administración & dosificaciónRESUMEN
Sarin (C(4)H(10)FO(2)P,O-isopropyl methylphosphonofluoridate) is a colourless, odourless and highly toxic phosphonate that acts as a cholinesterase inhibitor and disrupts neuromuscular transmission. Sarin and related phosphonates are chemical warfare agents, and there is a possibility of their application in a military or terrorist attack. This paper reports a lab-on-a-chip device for detecting a trace amount of sarin in a small volume of blood. The device should allow early detection of sarin exposure during medical triage to differentiate between those requiring medical treatment from mass psychogenic illness cases. The device is based on continuous-flow microfluidics with sequential stages for lysis of whole blood, regeneration of free nerve agent from its complex with blood cholinesterase, protein precipitation, filtration, enzyme-assisted reaction and optical detection. Whole blood was first mixed with a nerve gas regeneration agent, followed by a protein precipitation step. Subsequently, the lysed product was filtered on the chip in two steps to remove particulates and fluoride ions. The filtered blood sample was then tested for trace levels of regenerated sarin using immobilised cholinesterase on the chip. Activity of immobilised cholinesterase was monitored by the enzyme-assisted reaction of a substrate and reaction of the end-product with a chromophore. Resultant changes in chromophore-induced absorbance were recorded on the chip using a Z-shaped optical window. Loss of enzyme activity obtained prior and after passage of the treated blood sample, as shown by a decrease in recorded absorbance values, indicates the presence of either free or regenerated sarin in the blood sample. The device was fabricated in PMMA (polymethylmethacrylate) using CO(2)-laser micromachining. This paper reports the testing results of the different stages, as well as the whole device with all stages in the required assay sequence. The results demonstrate the potential use of a field-deployable hand-held device for point-of-care triage of suspected nerve agent casualties.
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Técnicas Biosensibles/métodos , Sustancias para la Guerra Química/análisis , Inhibidores de la Colinesterasa/sangre , Procedimientos Analíticos en Microchip/métodos , Sarín/sangre , Técnicas Biosensibles/instrumentación , Precipitación Química , Colinesterasas/química , Colinesterasas/metabolismo , Enzimas/química , Enzimas/metabolismo , Filtración , Humanos , Dispositivos Laboratorio en un Chip , Óptica y Fotónica , Proteínas/química , Proteínas/metabolismo , Factores de TiempoRESUMEN
A rapid, sensitive and robust immunoassay based on a commercial surface plasmon resonance (SPR) instrument (Biacore X) was developed for the detection of ricin in environmental samples. A total of 10 monoclonal antibodies were evaluated for their ability to recognise both a commercial ricin and horticultural ricin variants extracted from six different cultivars of Ricinus communis. Two suitable antibodies (7G12 and TFTA) were identified because of their strong affinity to all six ricin variants. The antibody 7G12 was used as the capture ligand in the SPR system. The assay was linear over a wide range of ricin concentrations (up to at least 750 ng/ml) with a limit of detection of 0.5 ng/ml. The assay was highly reproducible (coefficient of variation was less than 5%), and was able to detect all six ricin variants and environmental samples.
Asunto(s)
Monitoreo del Ambiente/métodos , Ricina/análisis , Resonancia por Plasmón de Superficie/métodos , Toxinas Biológicas/análisis , Anticuerpos Monoclonales , Ensayo de Inmunoadsorción Enzimática , Reproducibilidad de los ResultadosRESUMEN
A novel liquid chromatography-multiple reaction monitoring (LC-MRM) procedure has been developed for retrospective diagnosis of exposure to different forms of mustard agents. This concise method is able to validate prior exposure to nitrogen mustards (HN-1, HN-2, and HN-3) or sulfur mustard (HD) in a single run, which significantly reduces analysis time compared to separate runs to screen for different mustards' biomarkers based on tandem mass spectrometry. Belonging to one of the more toxic classes of chemical warfare agents, these potent vesicants bind covalently to the cysteine-34 residue of human serum albumin. This results in the formation of stable adducts whose identities were confirmed by a de novo sequencing bioinformatics software package. Our developed technique tracks these albumin-derived adduct biomarkers in blood samples which persist in vitro following exposure, enabling a detection limit of 200 nM of HN-1, 100 nM of HN-2, 200 nM of HN-3, or 50 nM of HD in human blood. The CWA-adducts formed in blood samples can be conveniently and sensitively analyzed by this MRM technique to allow rapid and reliable screening.
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Cromatografía Liquida/métodos , Monitoreo del Ambiente/métodos , Compuestos de Mostaza/análisis , Espectrometría de Masas en Tándem/métodos , Alquilación , Biomarcadores/sangre , Cromatografía de Afinidad , Biología Computacional/métodos , Exposición a Riesgos Ambientales/análisis , Humanos , Mecloretamina/análisis , Mecloretamina/metabolismo , Compuestos de Mostaza/metabolismo , Gas Mostaza/análisis , Gas Mostaza/metabolismo , Compuestos de Mostaza Nitrogenada/análisis , Compuestos de Mostaza Nitrogenada/metabolismo , Reproducibilidad de los Resultados , Albúmina Sérica/química , Albúmina Sérica/aislamiento & purificación , Albúmina Sérica/metabolismo , Programas Informáticos , Extracción en Fase SólidaRESUMEN
Animal experimental studies indicate that acute or chronic low-dose ionizing radiation (LDIR) (≤100 mSv) or low-dose-rate ionizing radiation (LDRIR) (<6 mSv/h) exposures may be harmful. It induces genetic and epigenetic changes and is associated with a range of physiological disturbances that includes altered immune system, abnormal brain development with resultant cognitive impairment, cataractogenesis, abnormal embryonic development, circulatory diseases, weight gain, premature menopause in female animals, tumorigenesis and shortened lifespan. Paternal or prenatal LDIR/LDRIR exposure is associated with reduced fertility and number of live fetuses, and transgenerational genomic aberrations. On the other hand, in some experimental studies, LDIR/LDRIR exposure has also been reported to bring about beneficial effects such as reduction in tumorigenesis, prolonged lifespan and enhanced fertility. The differences in reported effects of LDIR/LDRIR exposure are dependent on animal genetic background (susceptibility), age (prenatal or postnatal days), sex, nature of radiation exposure (i.e. acute, fractionated or chronic radiation exposure), type of radiation, combination of radiation with other toxic agents (such as smoking, pesticides or other chemical toxins) or animal experimental designs. In this review paper, we aimed to update radiation researchers and radiologists on the current progress achieved in understanding the LDIR/LDRIR-induced bionegative and biopositive effects reported in the various animal models. The roles played by a variety of molecules that are implicated in LDIR/LDRIR-induced health effects will be elaborated. The review will help in future investigations of LDIR/LDRIR-induced health effects by providing clues for designing improved animal research models in order to clarify the current controversial/contradictory findings from existing studies.
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Modelos Animales , Radiación Ionizante , Animales , Carcinogénesis/patología , Relación Dosis-Respuesta en la Radiación , Humanos , Plasticidad Neuronal/efectos de la radiaciónRESUMEN
Irradiation of the brain in early human life may set abnormal developmental events into motion that last a lifetime, leading to a poor quality of life for affected individuals. While the effect of irradiation at different early developmental stages on the late human life has not been investigated systematically, animal experimental studies suggest that acute postnatal irradiation with ⩾0.1Gy may significantly reduce neurogenesis in the dentate gyrus and endotheliogenesis in cerebral vessels and induce cognitive impairment and aging. Fractionated irradiation also reduces neurogenesis. Furthermore, irradiation induces hippocampal neuronal loss in CA1 and CA3 areas, neuroinflammation and reduces gliogenesis. The hippocampal neurovascular niche and the total number of microvessels are also changed after radiation exposures. Each or combination of these pathological changes may cause cognitive impairment and aging. Interestingly, acute irradiation of aged brain with a certain amount of radiation has also been reported to induce brain hormesis or neurogenesis. At molecular levels, inflammatory cytokines, chemokines, neural growth factors, neurotransmitters, their receptors and signal transduction systems, reactive oxygen species are involved in radiation-induced adverse effect on brain development and functions. Further study at different omics levels after low dose/dose rate irradiation may not only unravel the mechanisms of radiation-induced adverse brain effect or hormesis, but also provide clues for detection or diagnosis of radiation exposure and for therapeutic approaches to effectively prevent radiation-induced cognitive impairment and aging. Investigation focusing on radiation-induced changes of critical brain development events may reveal many previously unknown adverse effects.
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Envejecimiento/efectos de la radiación , Disfunción Cognitiva/etiología , Hipocampo/efectos de la radiación , Traumatismos por Radiación/patología , Traumatismos por Radiación/psicología , Envejecimiento/patología , Envejecimiento/fisiología , Animales , Disfunción Cognitiva/patología , Disfunción Cognitiva/fisiopatología , Hipocampo/patología , Hipocampo/fisiopatología , Humanos , Neurogénesis/fisiología , Neurogénesis/efectos de la radiación , Traumatismos por Radiación/fisiopatologíaRESUMEN
The effect of acute irradiation with 5Gy or fractionated exposure with 0.5Gy continuously for 10days (a total dose of 5Gy) was evaluated in an immature BALB/c mouse model. Radioprotective effect of ursolic acid (at 25mg/kg/daily administered 1h after acute or each of fractionated irradiations, and continuously for 30days) was also investigated. We found that both acute and fractionated irradiation at a total dose of 5Gy did not induce any mortality within 30days after exposure to postnatal day 26 (P26) BALB/c mice, but reduced animal weigh gain in the first few weeks. At 90days after irradiation, the weight of animals with acute irradiation was still significantly lower than the control group; no significant difference though was observed for those fractionatedly exposed mice compared to the control group. Behavioral tests indicated that acute irradiation at 5Gy induced deficits in learning and memory in the contextual fear conditioning test. The memory for novel object recognition was also impaired. Similar changes were not observed in mice with fractionated irradiation. Immunohistochemical study demonstrated clearly that acute and fractionated irradiations induced impairment of neurogenesis in the subgranular zone (SGZ) of the dentate gyrus although fractionated exposure induced much lesser loss of newly generated neurons. Ursolic acid administered at 25mg/kg/daily for 30days after irradiation greatly improved acute irradiation-induced deficits in contextual learning and memory and in novel object recognition memory although it exacerbated radiation-induced reduction of neurogenesis in SGZ.
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Inhibidores de la Ciclooxigenasa/uso terapéutico , Discapacidades para el Aprendizaje/tratamiento farmacológico , Discapacidades para el Aprendizaje/etiología , Neurogénesis/efectos de los fármacos , Traumatismos Experimentales por Radiación/complicaciones , Triterpenos/uso terapéutico , Animales , Animales Recién Nacidos , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Condicionamiento Psicológico/efectos de los fármacos , Condicionamiento Psicológico/efectos de la radiación , Modelos Animales de Enfermedad , Proteínas de Dominio Doblecortina , Miedo/efectos de los fármacos , Miedo/efectos de la radiación , Suspensión Trasera , Hipocampo/efectos de los fármacos , Hipocampo/efectos de la radiación , Antígeno Ki-67/metabolismo , Locomoción/efectos de los fármacos , Locomoción/efectos de la radiación , Ratones , Ratones Endogámicos BALB C , Proteínas Asociadas a Microtúbulos/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Neuropéptidos/metabolismo , Traumatismos Experimentales por Radiación/tratamiento farmacológico , Reconocimiento en Psicología/efectos de la radiación , Natación/psicología , Ácido UrsólicoRESUMEN
This study was aimed to evaluate the possible protective effects of ursolic acid (UA) against gamma radiation induced damage both in vitro as well as in vivo. It was observed that the exposure to gamma radiation dose- and time-dependently caused a significant decrease in the cell viability, while the treatment of UA attenuated this cytotoxicity. The production of free radicals including reactive oxygen species (ROS) and NO increased significantly post-irradiation and further induced lipid peroxidation and oxidative DNA damage in cells. These deleterious effects could also be effectively blocked by UA treatment. In addition, UA also reversed gamma irradiation induced inflammatory responses, as indicated by the decreased production of TNF-α, IL-6, and IL-1ß. NF-κB signaling pathway has been reported to be a key mediator involved in gamma radiation-induced cellular damage. Our results further demonstrated that gamma radiation dose- and time-dependently enhanced NF-κB DNA binding activity, which was significantly attenuated upon UA treatment. The post-irradiation increase in the expression of both phospho-p65, and phospho-IκBα was also blocked by UA. Moreover, the treatment of UA was found to significantly prolong overall survival in mice exposed to whole body gamma irradiation, and reduce the excessive inflammatory responses. Given its radioprotective efficacy as described here, UA as an antioxidant and NF-κB pathway blocker, may function as an important pharmacological agent in protecting against gamma irradiation-induced injury.
RESUMEN
Pharmacological properties of oxime reactivators, not related to its ability to regenerate or reactivate nerve agent-inhibited acetylcholinesterase located at nerve synapses, have been reported to be important in protecting against poisoning by the nerve agent soman. Such non-reactivation effects have thus far been associated only with bispyridinium oximes. This study investigated the possibility of creating similar non-reactivation therapeutic effects in the mono-pyridinium ring oxime, pralidoxime (2-PAM) through attachment of alkyl groups of increasing chain length to the oxime functional group. Of the 4 derivatives investigated, only the O-benzyl derivative displayed strong sedative effects in mice and mitigated the development of motor convulsions following soman challenge (1.8 x LD50, subcutaneous). Anticonvulsant effects of this compound were enhanced by co-administration of a non-anticonvulsant dose of atropine sulfate. Administration of equivalent amount of other O-derivatives of pralidoxime failed to elicit similar anticonvulsant actions. Electroencephalographic (EEG) and histopathological studies using the rat model, intoxicated with a lethal dose (1.6 x LD50, s.c.) of soman, confirmed O-benzyl derivative neuroprotective capabilities when used as a pretreatment drug. Microdialysis studies revealed that its neuroprotective effect is related to its ability to attenuate soman-induced increase in acetylcholine.
Asunto(s)
Derivados del Benceno/farmacología , Fármacos Neuroprotectores/farmacología , Compuestos de Pralidoxima/farmacología , Soman/toxicidad , Ácido 3,4-Dihidroxifenilacético/metabolismo , Acetilcolina/metabolismo , Análisis de Varianza , Animales , Anticonvulsivantes/farmacología , Atropina/farmacología , Derivados del Benceno/química , Inhibidores de la Colinesterasa/toxicidad , Reactivadores de la Colinesterasa/farmacología , Cuerpo Estriado/efectos de los fármacos , Cuerpo Estriado/metabolismo , Dopamina/metabolismo , Sinergismo Farmacológico , Electroencefalografía/efectos de los fármacos , Femenino , Ácido Homovanílico/metabolismo , Ácido Hidroxiindolacético/metabolismo , Ratones , Microdiálisis , Oximas , Compuestos de Pralidoxima/química , Compuestos de Piridinio/química , Compuestos de Piridinio/farmacología , Ratas , Ratas Wistar , Convulsiones/mortalidad , Convulsiones/fisiopatología , Convulsiones/prevención & control , Serotonina/metabolismo , Tasa de Supervivencia , Factores de TiempoRESUMEN
PURPOSES: To review research progress on the molecular mechanisms of low dose ionizing radiation (LDIR)-induced hormesis, adaptive responses, radioresistance, bystander effects, and genomic instability in order to provide clues for therapeutic approaches to enhance biopositive effects (defined as radiation-induced beneficial effects to the organism), and control bionegative effects (defined as radiation-induced harmful effects to the organism) and related human diseases. CONCLUSIONS: Experimental studies have indicated that Ataxia telangiectasia-mutated (ATM), extracellular signal-related kinase (ERK), mitogen-activated protein kinase (MAPK), phospho-c-Jun NH(2)-terminal kinase (JNK) and protein 53 (P53)-related signal transduction pathways may be involved in LDIR-induced hormesis; MAPK, P53 may be important for adaptive response; ATM, cyclooxygenase-2 (COX-2), ERK, JNK, reactive oxygen species (ROS), P53 for radioresistance; COX-2, ERK, MAPK, ROS, tumor necrosis factor receptor alpha (TNFα) for LDIR-induced bystander effect; whereas ATM, ERK, MAPK, P53, ROS, TNFα-related signal transduction pathways are involved in LDIR-induced genomic instability. These results suggest that different manifestations of LDIR-induced cellular responses may have different signal transduction pathways. On the other hand, LDIR-induced different responses may also share the same signal transduction pathways. For instance, P53 has been involved in LDIR-induced hormesis, adaptive response, radioresistance and genomic instability. Current data therefore suggest that caution should be taken when designing therapeutic approaches using LDIR to induce beneficial effects in humans.
Asunto(s)
Inmunidad Adaptativa/efectos de la radiación , Efecto Espectador/efectos de la radiación , Inestabilidad Genómica/efectos de la radiación , Hormesis/efectos de la radiación , Tolerancia a Radiación/efectos de la radiación , Animales , Relación Dosis-Respuesta en la Radiación , HumanosRESUMEN
ACE inhibitors and ARBs (angiotensin receptor blockers) have been shown to attenuate radiation injuries in animal models of lethal gamma irradiation. These two classes of drug act by curtailing the actions of angiotensin II-linked inflammatory pathways that are up-regulated during gamma radiation in organ systems such as the brain, lung, kidney, and bone marrow. ACE inhibitors inhibit ACE and attenuate the formation of angiotensin II from angiotensin I; ARBs block the angiotensin AT1 receptor and attenuate the actions of angiotensin II that are elicited through the receptor. DAA-I (des-aspartate-angiotensin I), an orally active angiotensin peptide, also attenuates the deleterious actions of angiotensin II. It acts as an agonist on the angiotensin AT1 receptor and elicits responses that oppose those of angiotensn II. Thus, DAA-I was investigated for its anticipated radioprotection in gamma irradiated mice. DAA-I administered orally at 800 nmole/kg/day for 30 days post exposure (6.4 Gy) attenuated the death of mice during the 30-day period. The attenuation was blocked by losartan (50 nmole/kg/day, i.p.) that was administered sequential to DAA-I administration. This shows that the radioprotection was mediated via the angiotensin AT1 receptor. Furthermore, the radioprotection correlated to an increase in circulating PGE2 of surviving animals, and this suggests that PGE2 is involved in the radioprotection in DAA-I-treated mice. At the hematopoietic level, DAA-I significantly improved two syndromes of myelosuppression (leucopenia and lymphocytopenia), and mice pre-treated with DAA-I prior to gamma irradiation showed significant improvement in the four myelodysplastic syndromes that were investigated, namely leucopenia, lymphocytopenia, monocytopenia and thrombocytopenia. Based on the known ability of PGE2 to attenuate the loss of functional hematopoietic stem and progenitor cells in radiation injury, we hypothesize that PGE2 mediated the action of DAA-I. DAA-I completely attenuated the increase in circulating level of two inflammatory cytokines, TNFα and IL-6, in irradiated mice; and this shows that DAA-I exerted additional anti-inflammatory actions, which could also have contributed to its radioprotection. These findings show that DAA-I acts via a novel mechanism of action on the angiotensin AT1 receptor to specifically release PGE2, which mediates radioprotection in the gamma irradiated mice.
Asunto(s)
Angiotensina I/análogos & derivados , Rayos gamma/efectos adversos , Síndromes Mielodisplásicos/tratamiento farmacológico , Protectores contra Radiación/farmacología , Angiotensina I/farmacología , Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Animales , Dinoprostona/metabolismo , Femenino , Interleucina-6/metabolismo , Losartán/farmacología , Ratones , Ratones Endogámicos BALB C , Síndromes Mielodisplásicos/metabolismo , Síndromes Mielodisplásicos/radioterapia , Receptor de Angiotensina Tipo 1/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
O-Substituted aldoximes of the cholinesterase reactivator pralidoxime (O-methyl 1, O-benzyl 2, O-propynyl 3 and O-butynyl 4 derivatives) were synthesized and found to exhibit strong binding affinities for muscarinic receptors in rat brain, heart and submandibulary glands. The aldoximes were noncompetitive antagonists of acetylcholine-induced contraction of the guinea pig ileum. A good correlation was observed between binding affinity and pK(B). Weak anticholinesterase activities were observed for these compounds. When given intracerebroventricularly into conscious rats before soman administration (0.9 LD(50), subcutaneously), the aldoximes, like atropine but not pralidoxime, protected against respiratory depression (3,4) and bradycardia (2). No protection against soman-induced pressor effects was noted. The protective effects of these aldoximes may be the outcome of compensatory mechanisms, of which the cholinergic receptor agonist and antagonist properties of these compounds may be important.
Asunto(s)
Reactivadores de la Colinesterasa/farmacología , Compuestos de Pralidoxima/farmacología , Acetilcolinesterasa/efectos de los fármacos , Acetilcolinesterasa/metabolismo , Animales , Atropina/farmacología , Unión Competitiva/efectos de los fármacos , Presión Sanguínea/efectos de los fármacos , Encéfalo/metabolismo , Carbacol/farmacología , Agonistas Colinérgicos/farmacología , Inhibidores de la Colinesterasa/farmacología , Relación Dosis-Respuesta a Droga , Femenino , Cobayas , Frecuencia Cardíaca/efectos de los fármacos , Íleon/efectos de los fármacos , Íleon/fisiología , Técnicas In Vitro , Masculino , Antagonistas Muscarínicos/farmacología , Contracción Muscular/efectos de los fármacos , Miocardio/metabolismo , Oximas/metabolismo , Oximas/farmacología , Compuestos de Pralidoxima/química , Compuestos de Pralidoxima/metabolismo , Ensayo de Unión Radioligante , Ratas , Ratas Sprague-Dawley , Receptores Muscarínicos/metabolismo , Respiración/efectos de los fármacos , Soman/farmacología , Relación Estructura-Actividad , Glándula Submandibular/metabolismoRESUMEN
L6 skeletal muscle cells overexpressed ICAM-1 when treated with H2O2. Maximum effect was observed at 200 µM H2O2. Des-aspartate-angiotensin I (DAA-I) concentration-dependently attenuated the overexpression. Maximum attenuation occurred at 10(-10) M DAA-I. H2O2 activated NFκB and its translocation into the nucleus of L6 muscle cells suggesting that NFκB mediates the H2O2-induced overexpression of ICAM-1. DAA-I inhibited the activation and translocation of NFκB. H2O2 is a major oxidant formed during skeletal muscle contraction and is implicated in oxidative stress and skeletal muscle damage in excessive unaccustomed exercise. The data show that DAA-I has antioxidant action, and its action was further investigated in the soleus muscle of mice subjected to 240 min of eccentric exercise on a rodent treadmill. The eccentric exercise induced superoxide formation and overexpression of ICAM-1 in the soleus muscle of the mice at 3 days post exercise. DAA-I (0.2 nmole/kg/day) administered orally on day 1 (pre-exercise) and 2 days post-exercise attenuated both the ROS formation and ICAM-1 overexpression. Earlier studies show that DAA-I acts as an agonist on the angiotensin AT1 receptor and elicits responses opposing those of angiotensin II. The present and earlier findings support the recent suggestion that angiotensin II is involved in skeletal muscle damage, and curtailment of its actions via ACE inhibitors and losartan protects and improves skeletal muscle damage. These findings open up new avenues for treatment and management of skeletal muscle damage via the interventions of the renin angiotensin system.