RESUMEN
AIM: To examine the influence of ultraviolet C (UVC) radiation on blood, saliva, semen, and naked DNA samples for preventing DNA cross-contamination on working surfaces in laboratories. METHODS: Blood, saliva, semen, and DNA isolated from buccal swab samples were obtained from a single male donor and applied to the laboratory working surfaces. UVC radiation was applied to these diluted and undiluted samples with or without previous decontamination of the working surfaces with 10% sodium hypochlorite and 20% ethanol. Genomic DNA was extracted using Chelex. After quantification, DNA was amplified using the AmpFlSTR® NGM™ PCR Amplification Kit. We tested and statistically analyzed DNA concentration, UVC dose, sample volume, radiation time, the number of correctly detected alleles on genetic loci, and the number of correctly detected alleles in four groups in which 16 loci were divided. RESULTS: When working surfaces were not decontaminated and were treated only with UVC radiation in the laboratory, the genetic profile for naked DNA could not be obtained after 2 minutes of UVC radiation and for saliva after 54 hours. For blood and semen, a partial genetic profile was obtained even after 250 hours of UVC radiation in the laminar. When working surfaces were decontaminated with 10% sodium hypochlorite and 20% ethanol, genetic profile could not be obtained for naked DNA after 2 minutes, for saliva after 4 hours, for blood after 16 hours, and for semen after 8 hours of UVC radiation in the laboratory. CONCLUSION: It is recommended to carefully and thoroughly clean working surfaces with 10% sodium hypochlorite and 20% ethanol followed by minimal 16-hour UVC exposure (dose approximately 4380 mJ/cm2) for complete and successful decontamination.
Asunto(s)
Sangre/efectos de la radiación , ADN/efectos de la radiación , Saliva/efectos de la radiación , Semen/efectos de la radiación , Rayos Ultravioleta , Daño del ADN , Amplificación de Genes , Humanos , Masculino , Reacción en Cadena de la Polimerasa , Dosis de Radiación , Adulto JovenRESUMEN
OBJECTIVE: The objective of this study was determination of causative factors of the genital infections and their correlation with various predictor variables. Secondary objectives included: (1) determination of the presence and the type of low molecular weight metabolites in the samples of vaginal secretion formed in vivo, (2) determination of the concentration of 2-phenylethanol formed in vitro for each Candida species, (3) determination of the relationship between fungal/bacterial/viral infections with the metabolites formed in vivo using multivariate analysis. METHODS: One hundred and ninety-seven women in the age range from 18 to 65 years were included in the study. After the completion of questionnaire, all the patients were subjected to Pap test, cervical swabs for the presence of aerobic bacteria, yeasts, Ureaplasma urealyticum, Chlamydia trachomatis, Mycoplasma, and hrHPV DNA. The presence and the concentration of low-molecular weight metabolites in vitro and in vivo were determined by gas chromatography-mass spectrometry (GC-MS) method. Multivariate analysis methods were used for statistical evaluation. RESULTS: The most important risk factors of fungal/bacterial/viral infections were determined. The presence of 2-phenylethanol in vivo was confirmed in 14 of 74 tested samples and connected with the Candida species. The presence of symptoms, hrHPV DNA and Ureaplasma urealyticum are the predictor variables with the highest influence on the formation of the metabolite in vivo. The results in vitro confirmed that various Candida species produced 2-phenylethanol with the concentrations ranging from 0.6 to 4.64 µg/mL. CONCLUSION: The medical exposure to irradiation, marital status, and number of partners as well as stress factors (miscarriages, chronic, viral, or tumor illnesses) had the highest influence on the development of the bacterial/fungal/viral infections. The formation of 2-phenylethanol, both in vivo and in vitro, was confirmed and connected with Candida species. Besides, according to statistical tests, it seems that presence of symptoms, hrHPV DNA, and Ureaplasma urealyticum had also significant role on the formation of 2-phenylethanol in vivo.
Asunto(s)
Candida/metabolismo , Alcohol Feniletílico/análisis , Vagina/microbiología , Vaginitis/microbiología , Adolescente , Adulto , Anciano , Estudios de Casos y Controles , Croacia/epidemiología , Femenino , Estado de Salud , Humanos , Estilo de Vida , Persona de Mediana Edad , Frotis Vaginal , Vaginitis/epidemiología , Adulto JovenRESUMEN
Nerve agents are highly toxic organophosphorus (OP) compounds. They inhibit acetylcholinesterase (AChE), an enzyme that hydrolyses acetycholine (ACh) in the nervous system. Pathophysiological changes caused by OP poisonings are primarily the consequence of surplus ACh on cholinergic receptors and in the central nervous system. Standard treatment of OP poisoning includes combined administration of carbamates, atropine, oximes and anticonvulsants. In order to improve therapy, new compounds have been synthesised and tested. Tenocyclidine (TCP) and its adamantane derivative 1-[2-(2-thienyl)-2-adamantyl] morpholine (TAMORF) have shown interesting properties against soman poisoning. In this study, we developed a qualitative GC-MS method to measure elimination of TCP and TAMORF through rat urine in order to learn more about the mechanisms through which TCP protects an organism from OP poisoning and to determine the duration of this protective effect. GC-MS showed that six hours after treatment with TCP, rat urine contained only its metabolite 1-thienylcyclohexene, while urine of rats treated with TAMORF contained both TAMORF and its metabolites.
Asunto(s)
Adamantano/análogos & derivados , Cromatografía de Gases y Espectrometría de Masas , Morfolinas/farmacocinética , Piperidinas/farmacocinética , Tiofenos/farmacocinética , Adamantano/farmacocinética , Adamantano/orina , Animales , Masculino , Morfolinas/orina , Intoxicación por Organofosfatos , Fenciclidina/análogos & derivados , Piperidinas/orina , Ratas , Ratas Wistar , Tiofenos/orinaRESUMEN
This study aimed to evaluate the antidotal potency of tenocyclidine (TCP) that probably might protect acetylcholinesterase (AChE) in the case of organophosphate poisoning. TCP was tested alone as a pretreatment or in combination with atropine as a therapy in rats poisoned with (1/4) and (1/2) of LD(50) of soman. Possible genotoxic effects of TCP in white blood cells and brain tissue were also studied. Results were compared with previous findings on the adamantyl tenocyclidine derivative TAMORF. TCP given alone as pretreatment, 5 min before soman, seems to be superior in the protection of cholinesterase (ChE) catalytic activity in the plasma than in brain, especially after administration of the lower dose of soman. Plasma activities of the enzyme after a joint treatment with TCP and soman were significantly increased at 30 min (P<0.001) and 24 h (P=0.0043), as compared to soman alone. TCP and atropine, given as therapy, were more effective than TCP administered alone as a pretreatment. The above therapy significantly increased activities of the enzyme at 30 min (P=0.046) and 24 h (P<0.001), as compared to controls treated with (1/4) LD(50) of soman alone. Using the alkaline comet assay, acceptable genotoxicity of TCP was observed. However, the controversial role of TCP in brain protection of soman-poisoned rats should be studied further.
Asunto(s)
Piperidinas/farmacología , Soman/envenenamiento , Tiofenos/farmacología , Acetilcolinesterasa/metabolismo , Animales , Encéfalo/metabolismo , Inhibidores de la Colinesterasa/farmacología , Reactivadores de la Colinesterasa/metabolismo , Colinesterasas/metabolismo , Ensayo Cometa , Leucocitos/metabolismo , Masculino , Modelos Químicos , Mutágenos/farmacología , Fenciclidina/análogos & derivados , Ratas , Ratas WistarRESUMEN
Tenocyclidine-TCP showing a broad spectrum of pharmacological activity including antidotal effect in organophosphorus compounds poisoning, radioprotective and anticancer effects. We investigated in vitro interactions of TCP and its adamantane derivative--TAMORF with human erythrocyte acetylcholinesterase (AChE). Moreover, their genotoxicity and radioprotective activity on human white blood cells were studied using the alkaline comet assay, viability testing and the analysis of the structural chromosome aberrations. The tested compounds were found to be weak inhibitors of AChE, for TCP IC(50)=1 x 10(-5)M and for TAMORF IC(50)>1 x 10(-3)M, without reactivating and protective effects on AChE inhibited by soman. Results suggest that TCP modified by the replacement of the cyclohexyl ring with an adamantly ring and piperidine with morpholine group (TAMORF) have lower toxicity. Both compounds possess low cytotoxicity and radioprotective activity, but TAMORF also shows cell growth inhibitory effects. To clarify differences in their biological efficiency observed in vitro and in vivo, additional analyses are necessary. Since TAMORF was found to significantly inhibit cell growth and proliferation in vitro, it is reasonably to consider it as a source molecule promising for further modifications and development of more potent substances with antitumor properties rather then radioprotector or antidote in organophosphorus poisoning.
Asunto(s)
Adamantano/análogos & derivados , Reactivadores de la Colinesterasa/farmacología , Morfolinas/farmacología , Piperidinas/farmacología , Tiofenos/farmacología , Adamantano/farmacología , Supervivencia Celular/efectos de los fármacos , Inhibidores de la Colinesterasa/toxicidad , Aberraciones Cromosómicas/efectos de los fármacos , Ensayo Cometa , Eritrocitos/efectos de los fármacos , Eritrocitos/enzimología , Rayos gamma , Humanos , Técnicas In Vitro , Dosificación Letal Mediana , Leucocitos/efectos de los fármacos , Leucocitos/efectos de la radiación , Linfocitos/efectos de los fármacos , Mutágenos/toxicidad , Fenciclidina/análogos & derivados , Protectores contra Radiación/farmacología , Soman/antagonistas & inhibidores , Soman/toxicidad , Relación Estructura-Actividad , Azul de TripanoRESUMEN
Gamma-hydroxybutyrate (GHB) is a naturally occurring compound present in the brain and peripheral tissues of mammals. It is a minor metabolite and precursor of gamma-aminobutyric acid (GABA). Just as GABA, GHB is believed to play a role in neurotransmission. GHB was first synthesized in vitro in 1960, when it revealed depressive and hypnotic effects on the central nervous system. In 1960s it was used as an anaesthetic and later as an alternative to anabolic steroids, in order to enhance muscle growth. However, after it was shown that it caused strong physical dependence and severe side effects, GHB was banned. For the last fifteen years, GHB has been abused for its intoxicating effects such as euphoria, reduced inhibitions and sedation. Illicitly it is available as white powder or as clear liquid. Paradoxically GHB can easily be manufactured from its precursor gamma-butyrolactone (GBL), which has not yet been banned. Because of many car accidents and criminal acts in which it is involved, GHB has become an important object of forensic laboratory analysis. This paper describes gas and liquid chromatography, infrared spectroscopy, microscopy, colourimetry and nuclear magnetic resonance as methods for detection and quantification of GHB in urine and illicit products.