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BACKGROUND: To date, no attempt has been made to collate literature on the relationship between the social environmental impact of COVID-19 and erectile dysfunction. The aim of this explorative review was to assess and compare the prevalence of erectile dysfunction (ED) in male healthcare workers and males during the COVID-19 pandemic. METHODS: A systematic review of major databases from inception to February 2021 was conducted. Prevalence data were extracted, and a random-effects meta-analysis was undertaken. OUTCOMES: The pooled prevalence of ED amongst healthcare workers working in COVID-19 specific environments, and non-healthcare during the COVID-19 pandemic. RESULTS: Of 52 initial studies, six were included for the final analysis. The pooled prevalence of ED in healthcare workers working in a COVID-19 environment was 63.6% (95% CI 20.3-92.3%), and in non-healthcare workers during the COVID-19 pandemic was 31.9% (95% CI 19.5-47.6%). CONCLUSION: The prevalence of ED in healthcare workers working in COVID-19 environments was higher than representative samples and is of concern. Sexual health (and by extension, overall health), should be a priority when considering ways to care for this population. Considering the social environmental impact of COVID-19 on sexual health and in particular on ED, it is important to provide adequate psychological support systems and to promote quality of life with particular attention to sexual health.
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COVID-19/epidemiología , Disfunción Eréctil/epidemiología , SARS-CoV-2 , Medio Social , Adolescente , Adulto , COVID-19/terapia , Disfunción Eréctil/psicología , Personal de Salud/psicología , Humanos , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
OBJECTIVES: The aim was to analyse the overall and sex-specific associations between cannabis use and physical activity and sedentary behaviour. STUDY DESIGN: Cross-sectional analyses from the National Health and Nutrition Examination Survey (NHANES). METHODS: Data on cannabis use and leisure time physical activity and sedentary behaviour from NHANES cycles 2007-2008 to 2015-2016 were analysed. Multivariable regression models were carried out. RESULTS: About 15,822 participants were analysed (mean age ± standard error = 37.5 ± 0.19 years, range 20-59 years). Significantly higher odds were found for being active and ever used cannabis in the overall sample (odds ratio [OR] = 1.2, 95% confidence interval [CI]: 1.1-1.4) and in males (OR = 1.3, 95% CI: 1.1 to 1.5) and females (OR = 1.2, 95% CI: 1.0-1.4), respectively. In respective of sedentary behaviour, ever used cannabis was associated with higher odds of TV viewing ≥2 h/day in the overall sample (OR = 1.2, 95% CI: 1.0-1.4). However, this association was observed in males only (OR = 1.3, 95% CI: 1.1-1.6). Ever used cannabis was associated with total sitting time (beta-coefficient = 0.3, 95%CI: 0.1-0.4), which was more evident in females (beta-coefficient = 0.4, 95% CI: 0.1-0.6). CONCLUSIONS: Cannabis consumption was associated with higher levels of physical activity and sitting time. When intervening to reduce cannabis consumption in the US populations, it may be appropriate to promote physical activity and ensure physical activity is maintained once cannabis consumption is stopped.
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Ejercicio Físico , Abuso de Marihuana/epidemiología , Conducta Sedentaria , Adulto , Estudios Transversales , Femenino , Humanos , Masculino , Persona de Mediana Edad , Encuestas Nutricionales , Factores de Riesgo , Distribución por Sexo , Factores de Tiempo , Estados Unidos/epidemiología , Adulto JovenRESUMEN
OBJECTIVES: The relationship between consuming ≥2 servings of fruits and ≥3 servings of vegetables a day, which has been identified as optimal for health (i.e., adequate fruit/vegetable consumption), and non-communicable diseases (NCDs) in low- and middle-income countries (LMICs) is largely unknown. Therefore, using data from six LMICs, we investigated the independent association between inadequate fruit/vegetable consumption and 12 NCDs, and estimated the prevalence of inadequate fruit/vegetable consumption among people with NCDs. DESIGN AND SETTING: Cross-sectional, nationally representative data from the WHO Study on global AGEing and adult health (SAGE) were analyzed. PARTICIPANTS: Data on 34129 individuals aged ≥50 years were analyzed [mean (SD) age 62.4 (16.0); maximum age 114 years; 52.1% females]. MEASUREMENTS: Information on the number of servings of fruits and vegetables consumed on a typical day was self-reported. Twelve NCDs were assessed. Multivariable logistic regression analysis was conducted. RESULTS: Overall, 67.2% had inadequate fruit/vegetable consumption. Inadequate fruit/vegetable consumption was independently associated with significantly higher odds for chronic lung disease (OR=1.25), diabetes (OR=1.45), hearing problems (OR=1.75), and visual impairment (OR=2.50). The prevalence of inadequate fruit/vegetable consumption was particularly high among people with visual impairment (92.5%), depression (90.5%), asthma (79.8%), and hearing problems (78.4%). CONCLUSION: Promotion of fruit and vegetable consumption (≥2 servings of fruits and ≥3 servings of vegetables a day) in LMICs may lead to prevention of some NCDs (e.g., diabetes, chronic lung disease). Furthermore, people with certain NCDs (e.g., visual impairment, depression) had particularly high prevalence of inadequate fruit/vegetable consumption, and it is thus important to target this population to increase fruit/vegetable consumption.
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Enfermedades Pulmonares , Enfermedades no Transmisibles , Femenino , Humanos , Masculino , Frutas , Verduras , Enfermedades no Transmisibles/epidemiología , Países en Desarrollo , Estudios Transversales , Dieta , Trastornos de la VisiónRESUMEN
The cytochrome P450 system plays a key role in the metabolism of endogenous and exogenous compounds. The system is distributed widely in body tissues, with the highest concentration of the enzymes found in liver hepatocytes. Extrahepatic expression of the P450 system has been documented in the lung, pancreas and kidney, and the enzymes are induced by many disease states, including diabetes mellitus and cancer. Little attention has been paid to the expression and inducibility of the system in peripheral blood lymphocytes. In this study, specific P450 inducers are administered in vivo to male Wistar rats. The expression and in vivo induction of the P450 isoforms CYP2B, CYP2E, CYP3A and CYP4A in liver and lymphocyte samples is determined using Western blot analysis. Following in vivo induction, the lymphocyte P450 proteins showed an average three-fold increase in expression (0.003-0.005 microg P450/microg microsomal protein), compared to the control lymphocyte samples. Expression in the induced lymphocyte samples was up to 11-fold lower than that in the induced liver samples, as expected. These results indicate that lymphocytes may provide a relatively simple method by which to monitor the P450 profile in human subjects.
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Sistema Enzimático del Citocromo P-450/metabolismo , Hepatocitos/enzimología , Linfocitos/enzimología , Animales , Western Blotting , Células Cultivadas , Citocromo P-450 CYP2B1/metabolismo , Citocromo P-450 CYP2E1/metabolismo , Citocromo P-450 CYP3A/metabolismo , Citocromo P-450 CYP4A/metabolismo , Inducción Enzimática , Humanos , Masculino , Microsomas , Ratas , Ratas WistarRESUMEN
BACKGROUND: Antioxidant status can be used as a biomarker to assess chronic disease risk and diet can modulate antioxidant defence. OBJECTIVE: To examine effects of vegetarian diet and variations in the habitual intakes of foods and nutrients on blood antioxidants. SUBJECTS AND SETTING: Thirty-one vegetarians (including six vegans) and 58 omnivores, non-smokers, in Northern Ireland. DESIGN: A diet history method was used to assess habitual diet. Antioxidant vitamins, carotenoids, uric acid, zinc- and ferric-reducing ability of plasma (FRAP) were measured in fasting plasma and activities of glutathione peroxidase (GPX), superoxide dismutase (SOD) and glutathione S-transferase (GST) and level of reduced glutathione (GSH) were measured in erythrocytes. RESULTS: Vegetarians had approximately 15% higher levels of plasma carotenoids compared with omnivores, including lutein (P< or =0.05), alpha-cryptoxanthin P< or =0.05), lycopene (NS), alpha-carotene (NS) and beta-carotene (NS). The levels/activities of all other antioxidants measured were similar between vegetarians and omnivores. Total intake of fruits, vegetables and fruit juices was positively associated with plasma levels of several carotenoids and vitamin C. Intake of vegetables was positively associated with plasma lutein, alpha-cryptoxanthin, alpha-carotene and beta-carotene, whereas intake of fruits was positively associated with plasma beta-cryptoxanthin. Intake of tea and wine was positively associated with FRAP value, whereas intake of herbal tea associated positively with plasma vitamin C. Intakes of meat and fish were positively associated with plasma uric acid and FRAP value. CONCLUSIONS: The overall antioxidant status was similar between vegetarians and omnivores. Good correlations were found between intakes of carotenoids and their respective status in blood.
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Antioxidantes/administración & dosificación , Antioxidantes/metabolismo , Carotenoides/sangre , Dieta Vegetariana , Carne , Adulto , Biomarcadores/sangre , Carotenoides/administración & dosificación , Estudios de Cohortes , Eritrocitos/metabolismo , Conducta Alimentaria , Femenino , Frutas , Humanos , Peroxidación de Lípido/efectos de los fármacos , Masculino , Oxidación-Reducción , VerdurasRESUMEN
Germinating conidiospores (conidia) of Aspergillus nidulans amino acid-requiring strains are hypersensitive to heat, oxidative stress, UV radiation and chemical mutagens when compared with other strains. They also showed an increased mutation rate. Sensitivity to stress conditions has been correlated with an abnormal RAS/cAMP pathway in mutants of S. cerevisiae. We suggest that the RAS/cAMP pathway is defective in germinating conidia of Aspergillus amino acid auxotrophs and that this is responsible for suppressing DNA repair and conferring sensitivity to oxidative stress and heat shock.
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Aminoácidos/metabolismo , Aspergillus nidulans/metabolismo , 4-Nitroquinolina-1-Óxido/farmacología , Aspergillus nidulans/genética , Aspergillus nidulans/crecimiento & desarrollo , Daño del ADN , Calor , Mutación , Estrés Oxidativo , Esporas Fúngicas/metabolismo , Rayos UltravioletaRESUMEN
An ELISA used to investigate DNA repair in mammalian cells has been adapted to investigate mutagen-induced DNA damage and repair in protoplasts of Aspergillus nidulans. The assay shows a reduced rate of repair of DNA damage in methionine and arginine auxotrophs (methG and argB), which were shown previously to be hypersensitive to UV radiation and chemical mutagens. The assay also showed a considerably reduced ability to repair mutagen-induced damage in the uv-sensitive mutants uvsB and uvsH. The increased sensitivity of amino acids auxotrophs to mutagens is, therefore, correlated with a reduced capacity to repair mutagen-induced DNA damage.
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Arginina/metabolismo , Aspergillus nidulans/genética , Daño del ADN , Reparación del ADN , Ensayo de Inmunoadsorción Enzimática , Metionina/metabolismo , 4-Nitroquinolina-1-Óxido/farmacología , Aspergillus nidulans/efectos de los fármacos , Aspergillus nidulans/aislamiento & purificación , Aspergillus nidulans/efectos de la radiación , ADN de Hongos/efectos de los fármacos , ADN de Hongos/genética , ADN de Hongos/efectos de la radiación , ADN de Cadena Simple/genética , Metilmetanosulfonato/farmacología , Mutagénesis , Mutágenos/farmacología , Protoplastos , Rayos UltravioletaRESUMEN
The genetic material of our cells is susceptible to damage by a wide variety of extrinsic and intrinsic entities. The amount of genetic damage accumulated in vivo will depend upon an individual's ability to defend against and/or repair DNA damage. T cells in vivo have been shown to accumulate DNA damage and mutations over time. The accumulation of such genetic damage will occur in T cells possessing a 'naive' or a 'memory' phenotype. Since T cells are required to undergo extensive clonal expansion upon antigenic stimulation, DNA damage and mutations may result in: a failure of T cells to proliferate, because of DNA damage-mediated cell cycle arrest; decreased rates of proliferation, as a consequence of selection in vivo against cells carrying certain mutations and/or apoptosis, triggered by critical levels of DNA damage. Thus, when T cells, containing critical levels of genetic damage, are required to undergo rapid clonal expansion in the presence of antigen, insufficient numbers of T cells may be produced and so the immune response would be sub-optimal. In this paper the possible contribution of DNA damage and/or mutation to the age-related alterations in T cell-mediated immune responses will be discussed.
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Envejecimiento/inmunología , Daño del ADN , Mutación , Linfocitos T/inmunología , Envejecimiento/genética , Animales , Antioxidantes , Reparación del ADN , Humanos , Linfocitos , Nitrógeno , Especies Reactivas de Oxígeno/inmunologíaRESUMEN
T cells in vivo have been shown to accumulate DNA damage with age. To investigate the effects of DNA damage on T cell biology we have utilised an in vitro human CD4+ T cell clone model. Levels and types of DNA damage were determined in 11 independent T cell clones as a function of their in vitro lifespan. Increased levels of reactive oxygen species (ROS) induced DNA damage with increasing age were found in all clones analysed using a modified alkaline comet assay. T cell clones underwent apoptosis at the end of their lifespans. There were no consistent changes in the mRNA levels for the cyclin-dependent kinase inhibitors (CKI) p16, p21, and p27 during the clones' lifespans. It appears that the increased levels of ROS induced DNA damage in the T cells is not the major trigger of apoptosis, via the p53/p21 pathway. In addition, at the end of their lifespans, the T cell clones did not display the CKI phenotype reported for senescent cells (an increase in p16 and p21 levels). Thus, while the T cell clones appear sensitive to ROS-induced DNA damage, the molecular mechanisms through which this influences T cell dysfunction with age remains to be elucidated.
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Envejecimiento/metabolismo , Linfocitos T CD4-Positivos/metabolismo , Proteínas de Ciclo Celular/metabolismo , Inhibidor p16 de la Quinasa Dependiente de Ciclina/metabolismo , Ciclinas/metabolismo , Daño del ADN , Proteínas Supresoras de Tumor , Adulto , Anciano , Anciano de 80 o más Años , Envejecimiento/genética , Apoptosis , Linfocitos T CD4-Positivos/citología , Recuento de Células , Proteínas de Ciclo Celular/genética , División Celular , Fraccionamiento Celular , Células Cultivadas , Células Clonales , Inhibidor p16 de la Quinasa Dependiente de Ciclina/genética , Inhibidor p21 de las Quinasas Dependientes de la Ciclina , Inhibidor p27 de las Quinasas Dependientes de la Ciclina , Ciclinas/genética , ADN/metabolismo , Humanos , Persona de Mediana Edad , Oxidación-ReducciónRESUMEN
Carnosine (beta-alanyl-L-histidine), an abundant naturally-occurring dipeptide has been shown to exhibit anti-ageing properties towards cultured cells, possibly due in part to its antioxidant/free radical scavenging abilities. In this paper the results of an investigation on the effects of carnosine, at the physiological concentration of 20 mM, on oxidative DNA damage levels and in vitro lifespan in peripheral blood derived human CD4+ T cell clones are reported. Under the culture conditions used (20% O(2)) long term culture with carnosine resulted in a significant increase in the lifespan of a clone derived from a healthy young subject. No such extension was observed when a T cell clone from a healthy old SENIEUR donor was similarly cultured. Culture with carnosine from the midpoint of each clone's lifespan did not have any effect on longevity, independent of donor age. Oxidative DNA damage levels were measured in the clones at various points in their lifespans. Carnosine acted as a weak antioxidant, with levels of oxidative DNA damage being lower in T cells grown long term in the presence of carnosine. The possibility that carnosine might confer anti-ageing effects to T cells under physiological oxygen tensions would appear to be worthy of further investigation.
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Carnosina/farmacología , Daño del ADN , Estrés Oxidativo , Linfocitos T/efectos de los fármacos , Linfocitos T/fisiología , Células Cultivadas , Senescencia Celular/fisiología , Células Clonales , HumanosRESUMEN
Deterioration of the immune system with aging ("immunosenescence") is believed to contribute to morbidity and mortality in man due to the greater incidence of infection, as well as possibly autoimmune phenomena and cancer in the aged. Dysregulation of T cell function is thought to play a critical part in these processes. Factors contributing to T cell immunosenescence may include a) stem cell defects, b) thymus involution, c) defects in antigen presenting cells (APC), d) aging of resting immune cells, e) disrupted activation pathways in immune cells, f) replicative senescence of clonally expanding cells. This review aims to consider the current state of knowledge on the scientific basis for and potential clinical relevance of those factors in immunosenescence.
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Senescencia Celular/inmunología , Linfocitos T/inmunología , Envejecimiento/inmunología , Envejecimiento/metabolismo , Animales , Senescencia Celular/fisiología , Citocinas/biosíntesis , Humanos , Modelos Biológicos , Linfocitos T/fisiología , Timo/inmunología , Timo/fisiologíaRESUMEN
Deterioration of the immune system with aging ("immunosenescence") is believed to contribute to morbidity and mortality in man due to the greater incidence of infection, as well as possibly autoimmune phenomena and cancer in the aged. Dysregulation of T cell function is thought to play a critical part in these processes. Factors contributing to T cell immunosenescence may include a) stem cell defects, b) thymus involution, c) defects in antigen presenting cells (APC), d) aging of resting immune cells, e) disrupted activation pathways in immune cells, f) replicative senescence of clonally expanding cells. This review aims to consider the current state of knowledge on the scientific basis for and potential clinical relevance of those factors in immunosenescence.
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Envejecimiento/inmunología , Senescencia Celular/inmunología , Linfocitos T/inmunología , Animales , Células Presentadoras de Antígenos/fisiología , Antígenos CD28/fisiología , Células Clonales , Citocinas/metabolismo , Citocinas/fisiología , Hematopoyesis/fisiología , Humanos , Receptores de Antígenos de Linfocitos T/fisiología , Transducción de Señal , Timo/fisiopatologíaRESUMEN
A precondition for the chemotherapeutic treatment of a variety of virally-induced human diseases and malignant conditions is a highly selective interaction of the drug molecule to be used with it's biological target. To ensure the development of novel, effective drugs, it is essential that the biological target is well characterised with regard to it's structure and activity. Such characterisation relies upon adequate amounts of pure target being available. One of the most important enzymatic importers for antimetabolites is the enzyme thymidine kinase. In this article an in vitro protein expression system is described which facilitates the production of milligram amounts of pure and biologically active thymidine kinase, from a number of important biological sources. Results have shown that the in vitro produced enzyme has the exact biochemical propeties of the in vivo enzyme. Thus the in vitro protein expression system is an ideal vechicle to facilitate an in depth investigation of the enzyme's biological properties.
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To further investigate the possibility for retroviral involvement in the etiology of human breast cancer we processed peripheral blood monocytes and malignant breast tissue biopsies from 10 patients with breast cancer (infiltrating ductal carcinoma or infiltrating lobular carcinoma; ages 40-80 years) and 20 normal healthy women (with no evidence or family history of breast cancer. 10 age-matched controls and 10 women age 22-27 years) for the assay of the retroviral enzyme, reverse transcriptase, using an ELISA and for election microscopy examination for the detection of retroviral-like particles. Reverse transcriptase activity was detected in 5 out of 10 samples of monocyte culture medium and in 1 out of 10 of malignant tissue biopsies from the patients with breast cancer. In contrast, reverse transcriptase was not detected in the culture medium of the monocytes from any of the control subjects. Electron microscopy did not reveal the presence of any retroviral-like particles in any sample of monocyte culture medium or in any of the malignant or normal breast tissue biopsies. Despite evidence for the presence of reverse transcriptase in a subsample of the monocyte culture medium and breast tissue biopsies from the cohort of breast cancer patients who participated in this study, the role of retroviruses in human breast cancer remains unclear.
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Neoplasias de la Mama/microbiología , Infecciones por Retroviridae , Retroviridae/patogenicidad , Infecciones Tumorales por Virus , Adulto , Anciano , Neoplasias de la Mama/enzimología , Neoplasias de la Mama/ultraestructura , Femenino , Humanos , Microscopía Electrónica , Persona de Mediana Edad , ADN Polimerasa Dirigida por ARN/metabolismo , Infecciones por Retroviridae/enzimología , Infecciones Tumorales por Virus/enzimologíaRESUMEN
The mtDNA genome has been implicated as playing a pivotal role in determining the longevity and success of the human lifespan. A PCR-RFLP methodology was used to identify polymorphic restriction enzyme sites within a 2643 bp region of the mtDNA genome and a table of genetic haplotypes for a healthy aged and a younger control cohort of patients was constructed. Forty-six different mtDNA haplotypes and 11 groups of related haplotypes were identified across the two age groups but statistical analysis failed to show any significant associations. The European J haplogroup, previously reported to be associated with longevity, was not found at an increased frequency within the Irish aged population (P=0.36). However, the haplotypes comprising the J haplogroup could be differentiated into two distinct branches by the presence or absence of the two polymorphic restriction sites, 16,389g and 16,000g. The branch of haplotypes defined by 16,389g displayed a significant increased frequency in the aged samples (8%) compared to the controls (1%), P=0.015. Inversely, the branch of haplotypes defined by 16,000g displayed a significant decreased frequency in the aged samples (4%) compared to the controls (13%), P=0.011. The polymorphism (mt5178A) associated with longevity in the Japanese was not found in the Irish population, while the polymorphism (mt9055A) associated with successful ageing in the French centenarians was found at an increased frequency in the Irish aged population (9%) compared to the younger control group (5%), but failed to reach a level of statistical significance, P=0.164.
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ADN Mitocondrial/fisiología , Longevidad/genética , Polimorfismo Genético , Adulto , Anciano , Anciano de 80 o más Años , ADN Mitocondrial/clasificación , Europa (Continente) , Femenino , Haplotipos , Humanos , Irlanda , Masculino , Persona de Mediana Edad , FilogeniaRESUMEN
Nucleotide pool imbalances have been reported to affect the fidelity of DNA replication and repair in prokaryotic and eukaryotic cells. We have reported previously that the mutagen-hypersensitive thymidine kinase (TK)-deficient Friend erythroleukemia (FEL) cells (subclones 707BUF and 707BUE), have a more than sixfold increase in the dCTP:dTTP pool ratio when compared to that of wild-type, TK-positive (TK(+)) clone 707 cells. In this study we present the results of an investigation of the effect of the dCTP:dTTP pool imbalance on the accuracy of DNA replication within 707BUF cells. We examined the spontaneous mutation spectra occurring at the adenine phosphoribosyltransferase (aprt) locus within clone 707 (TK(+)) and 707BUF (TK(-)) FEL cells. Mutations recovered at the aprt locus in FEL cells comprised: base substitutions (43:73), frameshifts (14:13.5), and deletions (43:13.5) [clone 707 (TK(+)):707BUF (TK(-)), respectively, expressed as percentages]. A comparison of the mutation spectra obtained for the two cell lines did not reveal any significant increase in misincorporation of dCTP, the nucleotide in excess, in 707BUF (TK(-)) cells, during DNA replication synthesis. These data suggest that the dCTP:dTTP pool imbalance does not alter the fidelity of DNA replication synthesis in 707BUF (TK(-)) FEL cells. Rather, the predominance of GC --> AT transitions (53%) in the 707BUF (TK(-)) spectrum may reflect a reduced efficiency of repair by uracil DNA glycosylase of uracil residues within these cells.
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Replicación del ADN/genética , Nucleótidos de Desoxicitosina/metabolismo , Leucemia Eritroblástica Aguda/genética , Nucleótidos de Timina/metabolismo , Adenina Fosforribosiltransferasa/genética , Animales , Células Clonales , Análisis Mutacional de ADN/métodos , Nucleótidos de Desoxicitosina/genética , Exones , Leucemia Eritroblástica Aguda/metabolismo , Ratones , Mutación , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo Conformacional Retorcido-Simple , Análisis de Secuencia de ADN , Timidina Quinasa/deficiencia , Timidina Quinasa/genética , Nucleótidos de Timina/genética , Células Tumorales CultivadasRESUMEN
We are constantly exposed, throughout life, to a wide variety of extrinsic and intrinsic agents which have the potential to damage cellular biomolecules, including DNA. Imperfections in cellular defence systems which protect against the fixation of DNA damage can lead to an accumulation of mutations which on their own, or in combination with other age-related changes, may contribute to ageing and the development of age-related pathologies. We have previously reported an increase in frequency of mutation with age in human lymphocytes taken from healthy males in the age groups, 35-39, 50-54 and 65-69 years. In this article we report on the findings of a recent study which was designed to assess whether the age-related increase in frequency of mutation was due to a decreased efficacy of the defence systems against ROS-induced DNA damage, namely antioxidant status and DNA repair processes, in the same study subjects. In vivo antioxidant status was assessed in each of the study subjects by measuring blood levels of; superoxide dismutase (SOD; EC 1.15.1.1), glutathione peroxidase (GPx; EC 1.11.1.9), catalase (EC 1.11.1.6), caeruloplasmin (CPL), uric acid and bilirubin. We did not find any statistically significant differences in the mean levels of these antioxidants between the three different age groups. To investigate the efficacy of DNA repair processes against ROS-induced DNA damage, an ELISA was used to quantitate DNA damage (as % single-stranded DNA; %SS-DNA) at various times following treatment of peripheral blood lymphocytes with hydrogen peroxide (H2O2). The results of this part of the study showed that in untreated lymphocytes, basal levels of %SS-DNA were significantly higher in individuals from the 65-69 years age group compared to the 35-39 years age group (p = 0.039, 0.0013; at 5% level of significance). No significant differences were found in H2O2 susceptibility with age immediately following treatment (p = 0.71, 1.00; at 5% level of significance) but a consistent and significant increase was observed in %SS-DNA remaining 90 min post-treatment in lymphocytes from subjects in the 65-69 years age group, compared to %SS-DNA present in lymphocytes from the 35-39 years age group (p = 0.013, 0.024; at 5% level of significance). The results of this study suggest that the age-related increase in frequency of mutations is not contributed to by alterations of in vivo antioxidant status with age but is by a decreased efficacy of the repair of ROS-induced DNA damage with age. The biological implications of somatic mutations in the ageing process are discussed.
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Envejecimiento , Antioxidantes/metabolismo , Reparación del ADN , Mutación , Adulto , Anciano , Envejecimiento/genética , Envejecimiento/metabolismo , Bilirrubina/sangre , Catalasa/sangre , Ceruloplasmina/metabolismo , Daño del ADN/genética , ADN de Cadena Simple/genética , ADN de Cadena Simple/metabolismo , Eritrocitos/metabolismo , Glutatión Peroxidasa/sangre , Humanos , Peróxido de Hidrógeno/toxicidad , Linfocitos/metabolismo , Masculino , Persona de Mediana Edad , Especies Reactivas de Oxígeno/toxicidad , Superóxido Dismutasa/sangre , Ácido Úrico/sangreRESUMEN
The purpose of this study was to investigate the cytotoxic and genotoxic potential of low intensity laser irradiation (660 nm, 12 mW, 5 kHz) on mammalian cells. Thymidine kinase (TK)-positive and TK-deficient Friend erythroleukaemia (FEL) cells, clone 707 and subclone 707BUF respectively, were used in this investigation. Following irradiation of exponentially growing cells in suspension at doses of 2 and 20 J/cm2 a number of sensitive bioassays were used to facilitate the detection of laser-induced mutations, DNA damage and cell killing. Mutations were assessed by the examination of chromosome spreads, the determination of micronucleus frequency and by the determination of the mutant frequency at the hypoxanthine-guanine phosphoribosyltransferase (hgprt) locus. DNA damage was quantified using a sensitive ELISA. The cytotoxic effect of laser irradiation was assessed using a cloning assay. The results of this investigation did not show any significant increase in mutation frequency, DNA damage or cell survival in the laser-irradiated cells, compared to sham-irradiated controls. The lack of any demonstrable cytotoxic and genotoxic effects of low intensity laser irradiation on mammalian cells in culture would support it as being a safe modality for clinical use.
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Rayos Láser , Leucemia Eritroblástica Aguda/genética , Linfocitos/efectos de la radiación , Línea Celular , Supervivencia Celular , Daño del ADN , Virus de la Leucemia Murina de Friend , Humanos , Pruebas de Mutagenicidad , Células Tumorales CultivadasRESUMEN
Hyperbaric oxygen (HBO) treatment (i.e., exposure to 100% oxygen at a pressure of 2.5 atmosphere absolute (ATA) for a total of 3 x 20 min periods) of human subjects induced DNA damage in the alkaline comet assay with leukocytes and protected against the DNA damaging effects of subsequent in vivo HBO exposures. Furthermore, blood taken 24 h after the first HBO was well protected against the in vitro induction of genotoxic effects by hydrogen peroxide. To investigate the mechanisms which led to this apparent adaptive response, we determined the antioxidant status of blood from subjects before and after HBO. We did not find differences in the plasma concentrations of the antioxidant vitamins A, C and E after HBO treatment. HBO had also no effect on the 'antioxidant power' of the plasma as measured with the FRAP-assay or on the concentration of reduced glutathione determined in the plasma or in lymphocytes. Red cell concentrate activities of superoxide dismutase, catalase, glutathione peroxidase were not influenced by HBO. In contrast, synthesis of the heat shock protein HSP70 which has been implicated to play an important role in cellular protection against oxidative stress, was significantly induced in lymphocytes after a single HBO treatment. To investigate whether intake of antioxidants may protect against HBO-induced DNA damage, we supplemented subjects with vitamin E (800 mg for 7 days) or with N-acetylcysteine (400 mg, 1 h before the HBO treatment). However, these supplementations did not influence the induction of DNA damage by HBO.
Asunto(s)
Antioxidantes/metabolismo , Oxigenoterapia Hiperbárica/efectos adversos , Acetilcisteína/farmacología , Adaptación Fisiológica , Adulto , Ácido Ascórbico/sangre , Daño del ADN , Eritrocitos/metabolismo , Glutatión/sangre , Humanos , Peróxido de Hidrógeno/toxicidad , Técnicas In Vitro , Leucocitos/efectos de los fármacos , Leucocitos/metabolismo , Linfocitos/metabolismo , Vitamina A/sangre , Vitamina E/administración & dosificación , Vitamina E/sangreRESUMEN
An accumulation of mutations on their own or together with other age-related changes may contribute to aging and the development of age-related pathologies. The aim of this investigation was to assess the extent of DNA mutations as a function of age in humans. The mutant frequency (MF) at the hypoxanthine-guanine phosphoribosyl-transferase (hgprt) locus was assessed in lymphocytes isolated from male volunteers in each of three age groups (35-39, 50-54 and 65-69 years). Results show that the mean MF in the 65-69 years group was approximately twice that in the 35-39 and 50-54 years groups (4.1/10(6) cells, 1.9/10(6) cells and 1.79/10(6) cells, respectively) increasing by about 1.33% per year, after 54 years. In addition, there was an increased frequency of chromosomal aberrations in the 65-69 years group compared to the other two age groups. The results of this investigation show an increase in DNA mutations in cultured human lymphocytes with age. Factors which may influence the extent of DNA damage in human lymphocytes are discussed.