RESUMO
An experiment was conducted to compare bacterial counts of environmental mastitis pathogens in composted recycled manure solids bedding with those in fresh recycled manure solids. Eighteen Holstein cows were housed in 1 pen with 18 stalls. One row of 9 freestalls included mattresses and was bedded weekly with composted recycled manure solids. The second row of 9 freestalls included mattresses and was bedded weekly with fresh recycled manure solids. The back one-third of stalls toward the alleyway was covered in 25 to 50 mm of bedding. Samples were taken from the back one-third of 4 stalls for both treatments on d 0, 1, 2, and 6 of each week. After 3 wk, bedding treatments were switched between rows, making the total duration 6 wk. Mean total gram-negative bacterial counts were approximately 0.5 log10 cfu/g of dry matter lower in the composted recycled manure solids on d 0 compared with fresh recycled manure solids. Klebsiella species, coliform, and Streptococcus species counts were at least 1.0 log10 cfu/g of dry matter lower in composted compared with fresh recycled manure solids on d 0. Only gram-negative bacterial counts on d 1 were reduced in composted recycled manure solids compared with fresh recycled manure solids. Differences were not observed between treatments in gram-negative bacterial, coliform, Klebsiella species, or Streptococcus species counts on d 2 and 6. Ash content was higher in composted recycled manure solids compared with fresh recycled manure solids on d 0, 1, 2, and 6. Despite the increase in ash after composting, bacterial counts of mastitis pathogens in composted recycled manure solids were comparable with those in fresh recycled manure when used as freestall bedding.
Assuntos
Microbiologia Ambiental , Bactérias Gram-Negativas/isolamento & purificação , Esterco/microbiologia , Mastite Bovina/microbiologia , Streptococcus/isolamento & purificação , Animais , Carga Bacteriana/veterinária , Bovinos , Feminino , Abrigo para Animais , Klebsiella/isolamento & purificação , Reciclagem , SoloRESUMO
A watershed's water quality is influenced by contaminant-transport pathways unique to each landscape. Accurate information on contaminant-pathways could provide a basis for mitigation through well-targeted approaches. This study determined dynamics of nitrate-N, total P, Escherichia coli, and sediment during a runoff event in Tipton Creek, Iowa. The watershed, under crop and livestock production, has extensive tile drainage discharging through an alluvial valley. A September 2006 storm yielded 5.9 mm of discharge during the ensuing 7 d, which was monitored at the outlet (19,850 ha), two tile-drainage outfalls (total 1856 ha), and a runoff flume (11 ha) within the sloped valley. Hydrograph separations indicated 13% of tile discharge was from surface intakes. Tile and outlet nitrate-N loads were similar, verifying subsurface tiles dominate nitrate delivery. On a unit-area basis, tile total P and E. coli loads, respectively, were about half and 30% of the outlet's; their rapid, synchronous timing showed surface intakes are an important pathway for both contaminants. Flume results indicated field runoff was a significant source of total P and E. coli loads, but not the dominant one. At the outlet, sediment, P, and E. coli were reasonably synchronous. Radionuclide activities of (7)Be and (210)Pb in suspended sediments showed sheet-and-rill erosion sourced only 22% of sediment contributions; therefore, channel sources dominated and were an important source of P and E. coli. The contaminants followed unique pathways, necessitating separate mitigation strategies. To comprehensively address water quality, erosion-control and nitrogen-management practices currently encouraged could be complemented by buffering surface intakes and stabilizing stream banks.
Assuntos
Agricultura , Chuva , Movimentos da Água , Poluentes Químicos da Água/química , Água/química , Conservação dos Recursos Naturais , Monitoramento Ambiental , Escherichia coli/isolamento & purificação , Sedimentos Geológicos , Nitratos/química , Nitrogênio/química , Fósforo/química , Rios , Microbiologia da Água , Poluição Química da ÁguaRESUMO
BACKGROUND: Dietary heterocyclic amines (HCAs) are carcinogenic in rodent prostate requiring activation by enzymes such as cytochrome P450 (CYP) and N-acetyltransferase (NAT). METHODS: We investigated by Western blotting and immunohistochemistry the expression of CYP1A1, CYP1A2, and NAT1 in human prostate and in prostate epithelial cells (PECs) derived from primary cultures and tested their ability to activate the dietary carcinogen 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and its N-hydroxy metabolite (N-OH-IQ) to DNA-damaging moieties. RESULTS: Western blotting identified CYP1A1, CYP1A2, and NAT1. Immunohistochemistry localized NAT1 to the cytoplasm of PECs. Inter-individual variation was observed in the expression levels of CYP1A1, 1A2, and NAT1 (11, 75, and 35-fold, respectively). PECs expressed CYP1A1 and NAT1 but not CYP1A2. When incubated with IQ or N-OH-IQ, PECs formed DNA adducts indicating their ability to metabolically activate these compounds. CONCLUSIONS: Prostate cells possess the capacity to activate dietary carcinogens. PECs may provide a useful model system to study their role in prostate carcinogenesis.
Assuntos
Arilamina N-Acetiltransferase/análise , Citocromo P-450 CYP1A1/análise , Citocromo P-450 CYP1A2/análise , Células Epiteliais/enzimologia , Imidazóis/metabolismo , Isoenzimas/análise , Próstata/enzimologia , Quinolinas/metabolismo , Xenobióticos/metabolismo , Arilamina N-Acetiltransferase/genética , Arilamina N-Acetiltransferase/metabolismo , Biotransformação , Western Blotting , Carcinógenos/metabolismo , Células Cultivadas , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP1A2/genética , Citocromo P-450 CYP1A2/metabolismo , Adutos de DNA , DNA de Neoplasias/análise , Células Epiteliais/citologia , Regulação Enzimológica da Expressão Gênica , Humanos , Imuno-Histoquímica , Isoenzimas/genética , Isoenzimas/metabolismo , Masculino , Próstata/citologiaRESUMO
Differences in the incidence of prostate cancer (CaP) amongst different migrant populations point to causative agents of dietary and/or environmental origin. Prostate tissues were obtained following transurethral resection of the prostate (TURP) or radical retropubic prostatectomy. After surgery, TURP-derived or tumour-adjacent tissue fragments were minced in warm PFMR-4A medium (37 degrees C) and suspensions pipetted into collagen-coated petri dishes. Non-adherent material was removed by washing with fresh medium after 12 h. Adhered cells subsequently reacted positively with monoclonal antibodies to prostate specific antigen (PSA). PSA was also detected in the medium. The genotoxicities of the chemical carcinogens 2-amino-1-methyl-6-phenylimidazo[4,5-b] pyridine (PhIP), its N-hydroxy metabolite (N-OH-PhIP) and benzo[a]pyrene (B[a]P) in adherent cell populations from different donors (n=8) were examined. Cells were treated in suspension for 30 min at 37 degrees C in the presence of the DNA repair inhibitors hydroxyurea (HU) and cytosine arabinoside (ara-C). DNA single-strand breaks were detected in cells by the alkaline single cell-gel electrophoresis ('Comet') assay and quantified by measuring comet tail length (CTL) in microm. All three carcinogens induced dose-related increases in CTLs (P<0.0001) in cells from four donors 24 h post-seeding. However, in cells from a further two donors the genotoxic effects of PhIP, N-OH-PhIP and B[a]P were much less apparent after 48 h than after 24 h in culture. After 96 h in culture, cells from these donors appeared to be resistant to the comet-forming activity of the compounds. However, B[a]P-DNA adducts were still measurable by (32)P-postlabelling for up to 14 days following a 24-h exposure to 50 microM B[a]P in adhered cells from another two donors. This study shows that primary cultures of cells derived from the prostate can activate members of two classes of chemical carcinogens. Further development may provide a robust model system in which to investigate the aetiology of CaP.
Assuntos
Benzo(a)pireno/metabolismo , Benzo(a)pireno/farmacologia , Carcinógenos/metabolismo , Carcinógenos/farmacologia , Dano ao DNA , Imidazóis/metabolismo , Imidazóis/farmacologia , Neoplasias da Próstata/fisiopatologia , Piridinas/metabolismo , Piridinas/farmacologia , Adulto , Biotransformação , Ensaio Cometa , Sistema Enzimático do Citocromo P-450/farmacologia , Relação Dose-Resposta a Droga , Humanos , Masculino , Antígeno Prostático Específico , Células Tumorais CultivadasRESUMO
Breast cancer may be initiated by environmental/dietary agents and human milk may act as an ex vivo indicator of in vivo exposure of mammary epithelial cells to genotoxins. Extracts of human milk from UK-resident women (n=7) were tested for their abilities to morphologically transform C3H/M2 mouse fibroblasts. Genotoxicities were assessed in the Salmonella typhimurium reverse-mutation assay in the presence of S9 using strains TA1538 and YG1019, and in metabolically-competent human MCL-5 cells with the micronucleus and with the alkaline single cell gel electrophoresis (comet) assays. Two of the seven extracts were inactive in the transformation assay both in the presence or absence of S9, two appeared to be equally transforming either in the presence or absence of S9, and two other extracts induced increased transformation frequencies in the presence of S9. A seventh extract, tested only in the absence of S9, was inactive. Extracts were either active or inactive in at least three of the four tests applied. Four extracts were active or inactive in all four tests. The results suggest that human milk could be used as a resource for investigations of the as-yet-unidentified transforming agents previously detected in mammary lipid.
Assuntos
Fatores Biológicos/isolamento & purificação , Fatores Biológicos/toxicidade , Transformação Celular Neoplásica/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Leite Humano/química , Animais , Células Cultivadas , Fracionamento Químico , Cromatografia , Ensaio Cometa , Relação Dose-Resposta a Droga , Feminino , Fibroblastos/citologia , Humanos , Camundongos , Camundongos Endogâmicos C3H , Testes para Micronúcleos , Testes de Mutagenicidade , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/genética , Frações SubcelularesRESUMO
In this matched case-control study nested within the prospective Physicians' Health Study, we evaluated whether DNA damage in blood samples collected at enrollment significantly predicted risk, consistent with our hypothesis that cases have greater biological susceptibility to polycyclic aromatic hydrocarbons and other aromatic tobacco carcinogens. The subjects were 89 cases of primary lung cancer and 173 controls, all males, matched on smoking, age, and duration of follow-up. Aromatic-DNA adducts were measured in WBCs by the nuclease P1-enhanced (32)P-postlabeling method that primarily detects smoking-related adducts. Among current smokers, but not former or nonsmokers, there was a significant increase in mean adduct levels of cases compared with controls (11.04 versus 5.63; P = 0.03). "Healthy" current smokers who had elevated levels of aromatic DNA adducts in WBCs were approximately three times more likely to be diagnosed with lung cancer 1-13 years later than current smokers with lower adduct concentrations (odds ratio, 2.98; 95% confidence interval, 1.05-8.42; P = 0.04). We were not able to discern case-control differences in former smokers and nonsmokers. The findings are of interest because they suggest that individuals who become cases have greater biological susceptibility to tobacco carcinogens, a biological difference, which manifests most clearly while exposure is ongoing.
Assuntos
Carcinoma de Células Pequenas/sangue , Adutos de DNA/sangue , Dano ao DNA , Leucócitos/metabolismo , Neoplasias Pulmonares/sangue , Hidrocarbonetos Policíclicos Aromáticos/sangue , Carcinógenos/efeitos adversos , Carcinógenos/metabolismo , Carcinoma Pulmonar de Células não Pequenas/sangue , Carcinoma Pulmonar de Células não Pequenas/induzido quimicamente , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma de Células Pequenas/induzido quimicamente , Carcinoma de Células Pequenas/genética , Estudos de Casos e Controles , Humanos , Modelos Logísticos , Neoplasias Pulmonares/induzido quimicamente , Neoplasias Pulmonares/genética , Pessoa de Meia-Idade , Estudos Prospectivos , Fatores de RiscoRESUMO
Dietary and/or environmental factors appear to play a key role in the international variations that exist in breast cancer incidence. The genotoxicity of breast milk extracts is being examined as a possible indicator of in vivo exposure of mammary epithelial cells to DNA-damaging agents. Breast milk samples were obtained from the UK (n = 32), a high risk country, and from Hong Kong (n = 10), India (n = 20) and Singapore (n = 20), countries of lower breast cancer incidence. The abilities of breast milk extracts to induce DNA damage detected as single-strand breaks (SSBs) in the alkaline Comet assay and to induce micronuclei in MCL-5 cells and mutations in Salmonella typhimurium YG1019 were investigated. In the Comet assay 18 of 32 (56%) UK samples induced significant increases in DNA SSBs compared with 2 of 10 (20%), 5 of 20 (25%) and 8 of 20 (40%) of the samples from Hong Kong, India and Singapore, respectively. The proportion of positive samples was significantly higher in the UK group than in the combined low breast cancer incidence group and significantly higher than in the Indian group (P < 0.05, Fisher's exact test). In the micronucleus assay 9 of 32 (28%) UK samples showed significant activity compared with 0 of 10 (0%), 2 of 20 (10%) and 3 of 20 (15%) of the samples from Hong Kong, India and Singapore, respectively. Extracts of all the aforementioned milk samples were also tested for bacterial mutagenicity. Nine of 32 (28%) UK samples induced significant activity with a dose-response effect. Although activity was detected in samples from the other countries, comparable dose-response data could not be obtained because of a lack of material. This pilot study suggests that genotoxic components occur more frequently in UK breast milk than in milk from some other countries with a lower incidence of cancer. More work is required to confirm these initial findings and to examine their relevance to variations in breast cancer incidence.
Assuntos
Leite Humano/química , Mutagênicos/análise , Ensaio Cometa/métodos , Dano ao DNA/efeitos dos fármacos , Dano ao DNA/genética , Feminino , Humanos , Testes para Micronúcleos/métodos , Testes de Mutagenicidade , Mutagênicos/toxicidade , Projetos PilotoRESUMO
We investigated the effects of old age on the fingertip force responses that occurred when a grasped handle was pulled unexpectedly to increase the tangential load at the fingertip. These automatic responses, directed normal to the handle surface, help prevent slips between the handle and finger. Old adults (average age 78 years) responded with large peak fingertip forces compared to young adults (average age 30 years), even though the two subject groups showed similar skin slipperiness. For step-shaped loads the average response latency was the same for young and old subjects (about 80 ms). Thus, these automatic responses are not susceptible to the age-related central delays known for simple reaction-time tasks. For ramp-shaped loads the average response latency was inversely related to load rate. Response latency was 25 ms longer for the Old group versus the Young group for loads of 8 N/s, and this difference increased exponentially to a 110-ms difference for 2-N/s loads. A twofold difference in the tangential force required to evoke a response was predicted from linear regressions and can account for the latency difference (0.2 N vs 0.4 N threshold for young and old, respectively, r=0.93 for both groups). This theoretical elevation in load force threshold is consistent with degraded central information processing in old age, and the deterioration of cutaneous mechanoreceptors.
Assuntos
Envelhecimento/fisiologia , Força da Mão/fisiologia , Adulto , Idoso , Feminino , Humanos , Masculino , Neurônios Motores/fisiologia , Neurônios Aferentes/fisiologia , Tempo de Reação/fisiologia , Suporte de Carga/fisiologiaRESUMO
Exfoliated cells, isolated from breast milk samples donated by UK-resident women (n=15), were incubated, either immediately or after culture for 7 days, with one of a series of genotoxins, either in the presence or absence of the DNA-repair inhibitors, hydroxyurea (HU), and cytosine arabinoside (ara-C). The numbers of DNA single-strand breaks induced were then assessed as comet tail length (CTL) (microm) using the alkaline single cell-gel electrophoresis ('Comet') assay; cell viability was measured by trypan blue exclusion. The heterocyclic aromatic amines (HAAs) (2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) (0.4 mM), 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) (1.67 mM), 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) (1.77 mM)), a polycyclic aromatic hydrocarbon (benzo[a]pyrene (B[a]P) (0.36 mM)), a nitro-polycyclic aromatic hydrocarbon (1-nitropyrene (1-NP) (1.84 mM)) and aromatic amines (o-toluidine (0.85 mM), p-chloroaniline (0. 71 mM)) each induced statistically significant (P<0.0001, Mann-Whitney test) increases in median CTLs in breast milk cells from all the donors examined when incubated (30 min, 37 degrees C) in the presence of HU/ara-C. In some cases, these compounds were also active in the absence of the repair inhibitors. There were marked variations in comet formation between donors and between genotoxins. Cell culture appeared to increase the epithelial cell proportion and cultured cells retained their ability to activate genotoxins. The results suggest that breast milk is a valuable source of human mammary cells for the study of the metabolic activation of possible carcinogens.
Assuntos
Mama/metabolismo , Dano ao DNA , DNA/efeitos dos fármacos , Mutagênicos/farmacocinética , Biotransformação , Mama/citologia , Células Cultivadas , Ensaio Cometa , Células Epiteliais/metabolismo , Feminino , Humanos , Mutagênicos/toxicidadeRESUMO
Bis(dichloropropyl) ether isomers have been identified in a petrochemical plant effluent through a toxicity identification evaluation study in the United States. They have also been observed in the microgram per liter range along one of the largest rivers in Europe, the Elbe River. In the present investigation, the genotoxic and transforming activity of a bis(dichloropropyl) ether isomer, bis(2,3-dichloro-1-propyl) ether, was assayed in vitro. The results demonstrate that bis(2,3-dichloro-1-propyl) ether is a potent mutagen in Salmonella typhimurium strains TA 100, TA 1535, and to a lesser extent in strain TA 98, but only when tested in the presence of a metabolic activation system (S9 mix). We have also investigated the induction of micronuclei by bis(2,3-dichloro-1-propyl) ether in the metabolically competent cell line, MCL-5. A linear, dose-dependent increase in micronuclei was observed following exposure to bis(2,3-dichloro-1-propyl) ether. The DNA strand-breaking capacity of this chemical was assessed in the alkaline single-cell gel electrophoresis ("comet") assay with MCL-5 cells. Bis(2,3-dichloro-1-propyl) ether clearly induced DNA strand breaks in the 4.5-45.5 microg/ml dose range. The ether also induced malignant transformation in C3H/M2 mouse fibroblasts after metabolic activation (S9 mix). Thus, it must be suspected that bis(2, 3-dichloro-1-propyl) ether may possess a carcinogenic potential. Since the compound along with its isomers is present in considerable concentrations in surface water, their elimination is a matter of significant public concern.
Assuntos
Transformação Celular Neoplásica/efeitos dos fármacos , Éteres/toxicidade , Mutagênicos/toxicidade , Animais , Linhagem Celular , Camundongos , Camundongos Endogâmicos C3H , Testes para MicronúcleosRESUMO
Environmental and dietary factors are thought to be significant in breast cancer aetiology. The alkaline single-cell gel electrophoresis ('Comet') assay was used to examine breast milk cells for DNA damage and to measure the activity of extracts of the milk in causing such damage. UK-resident women were recruited as donors (n = 16) and provided 'early' ( approximately 4 weeks post-partum) and/or 'late' ( approximately 4 months post-partum) milk samples. Cells (79-94% viable, trypan blue exclusion) were either examined immediately for DNA damage or were cultured for 1 week prior to treatment with a breast milk extract. DNA damage in the form of single-strand breaks was quantified as comet tail length (CTL). Cell preparations examined immediately exhibited interindividual variation in median CTL (range 2.0-40.0 microm) with or without the DNA repair inhibitors hydroxyurea (HU) and cytosine arabinoside (ara-C). DNA damage decreased following culture, suggesting either DNA repair or death of DNA-damaged cells. Some donors' breast milk extracts induced DNA damage in their cultured cells and increases in median CTL were significantly greater with HU/ara-C (range 4.0-72.5 microm) than without (range 2.5-27.5 microm). Genotoxicity occurred without cytotoxicity (81-97% viability after treatment). Comparisons between cells and extracts from 'early' and 'late' milk samples did not support the idea of a progressive clearance of genotoxins from mammary lipid during lactation. Donors whose untreated cells contained the most DNA damage tended to yield genotoxic breast milk extracts. Cells isolated from milk activated the rodent mammary carcinogens o-toluidine and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). The relevance of genotoxic exposures to breast cancer initiation requires further investigation.
Assuntos
Neoplasias da Mama/etiologia , Dano ao DNA , Leite Humano/química , Mutagênicos/análise , Adulto , Ensaio Cometa , Feminino , Humanos , Leite Humano/citologiaRESUMO
Previous work has shown that a major route of activation of tamoxifen to DNA-binding products in rat liver cells is via alpha-hydroxylation leading to modification of the N(2)-position of guanine in DNA and to a lesser extent the N(6)-position of adenine. Improved resolution by HPLC has now identified two major adducts in rat liver DNA, one of them the aforementioned tamoxifen-N(2)-guanine adduct and the other the equivalent adduct in which the tamoxifen moiety has lost a methyl group. Treatment of rats or rat hepatocytes with N-desmethyltamoxifen gave rise to the second adduct, whereas treatment with tamoxifen or alpha-hydroxytamoxifen gave rise to both. Furthermore, N,N-didesmethyltamoxifen was found to be responsible for an additional minor DNA adduct formed by tamoxifen, alpha-hydroxytamoxifen and N-desmethyltamoxifen. The involvement of metabolism at the alpha position was confirmed in experiments in which [alpha-D(2)-ethyl]tamoxifen, but not [beta-D(3)-ethyl]tamoxifen, produced reduced levels of DNA adducts. Tamoxifen N-oxide and alpha-hydroxytamoxifen N-oxide also gave rise to DNA adducts in rat liver cells, but the adduct patterns were very similar to those formed by tamoxifen and alpha-hydroxytamoxifen, indicating that the N-oxygen is lost prior to DNA binding. These and earlier results demonstrate that in rat liver cells in vivo and in vitro, Phase I metabolic activation of tamoxifen involves both alpha-hydroxylation and N-demethylation, which is followed by Phase II activation at the alpha-position to form a highly reactive sulphate. Detection of tamoxifen-related DNA adducts by (32)P-postlabelling is achieved with >90% labelling efficiency.
Assuntos
Fígado/metabolismo , Tamoxifeno/farmacocinética , Animais , Biotransformação , Células Cultivadas , Cromatografia Líquida de Alta Pressão , DNA/metabolismo , Adutos de DNA , Feminino , Hidroxilação , Metilação , Oxirredução , Radioisótopos de Fósforo , Ratos , Ratos Endogâmicos F344RESUMO
We have found previously that the metabolically-competent human MCL-5 cell line did not appear to be usefully sensitive to the DNA-damaging effects of several carcinogens, as measured by the alkaline single-cell gel electrophoresis ('comet') assay. We therefore sought to increase its sensitivity by inhibiting DNA repair during exposure to test compounds, using 10 mM hydroxyurea (HU) and 1.8 mM cytosine arabinoside (ara-C), which inhibit DNA resynthesis during nucleotide excision repair. The following compounds were tested, using a 30-min exposure, in the absence or presence of HU/ara-C: 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (8-MeIQx), 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (4, 8-DiMeIQx), 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-amino-9H-pyrido[2,3-b]indole (A[alpha]C), 2-amino-3-methyl-9H-pyrido[2,3-b]indole (MeA[alpha]C), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), benzo[a]pyrene (B[a]P), 3-methylcholanthrene (3-MCA), 7, 12-dimethylbenz[a]anthracene (DMBA), 1-nitropyrene (1-NP), 2-nitrofluorene (2-NF), aniline, o-toluidine, benzene, lindane, bleomycin, cisplatin, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), sodium chromate, chromic chloride, and diethylstilboestrol (DES). We made the following observations. The background level of comet formation was reasonably constant over several months and was increased only slightly, but significantly, in the presence of the DNA-repair inhibitors. All compounds that induced comet formation did so without appreciable cytotoxicity as assessed by trypan blue exclusion. Of the compounds tested, the heterocyclic amines and polycyclic aromatic hydrocarbons (with the exceptions of PhIP and B[a]P) failed to induce convincing levels of comet formation in the absence of repair inhibitors. In their presence the heterocyclic amines tested induced comet formation (with the exception of 8-MeIQx), with widely differing potencies. 1-NP failed to elicit marked comet formation even in the presence of HU/ara-C. Aniline and o-toluidine produced significant levels of comet formation in the absence of HU/ara-C, but in their presence comet formation was markedly increased. Benzene, lindane, bleomycin, cisplatin, MNNG, sodium chromate and chromic chloride induced comet formation in the absence of HU/ara-C, but, with the exception of cisplatin, their presence enhanced comet formation. Neither sucrose nor DES elicited comet formation under the conditions used in this study. Many more agents need to be tested in order to determine how well the comet assay using MCL-5 cells (or modified versions of it) can distinguish genotoxins from non-genotoxins.
Assuntos
Citarabina/farmacologia , Reparo do DNA/efeitos dos fármacos , Eletroforese em Gel de Ágar/métodos , Hidroxiureia/farmacologia , Inibidores da Síntese de Ácido Nucleico/farmacologia , Aminas/toxicidade , Benzeno/toxicidade , Bleomicina/toxicidade , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Cloretos/toxicidade , Cromatos/toxicidade , Compostos de Cromo/toxicidade , Cisplatino/toxicidade , DNA/efeitos dos fármacos , DNA/genética , DNA/efeitos da radiação , Dano ao DNA , Dietilestilbestrol/toxicidade , Relação Dose-Resposta a Droga , Compostos Heterocíclicos/toxicidade , Hexaclorocicloexano/toxicidade , Humanos , Metilnitronitrosoguanidina/toxicidade , Testes de Mutagenicidade , Nitrocompostos/toxicidade , Hidrocarbonetos Policíclicos Aromáticos/toxicidade , Radiação Ionizante , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Compostos de Sódio/toxicidade , Sacarose/farmacologiaRESUMO
The CYP and GST genetic polymorphisms, controlling metabolism of xenobiotics, are considered to influence an individual's susceptibility to environmental and occupational carcinogens and predisposition to cancer. In the study, the effect of the GSTM1, GSTP1, CYP1A1 and CYP2D6 polymorphisms was investigated in relation to PAH-DNA adduct levels in non-tumourous lung tissue from non-small cell lung cancer (NSCLC) patients living in the industrialized region of Upper Silesia, Poland. The level of adducts among smokers was significantly elevated when compared to non-smokers (P = 0.0005). Adduct levels correlated inversely with age of patients (P = 0.00001). The GSTP1 and CYP2D6 polymorphisms had no influence on DNA adduct levels. There was a significant relationship between high adduct levels and the combined GSTM1 (null)/CYP1A1-Ile/Val genotype in the squamous cell carcinoma group (P = 0.028). An elevated number of adducts was found in patients with the GSTM1 (null)/CYP1Al-Ile/Val genotype compared to the GSTM1 (null)/CYP1A1-Ile/Ile carriers (P = 0.043). A higher frequency of the CYP1A1-Ile/Val and GSTM1 (null)/CYP1A1-Ile/Val genotypes was observed in patients with high adduct levels (P = 0.05 and P = 0.009, respectively). A significant prevalence of the GSTM1(null)/CYP1A1-Ile/Val carriers in the adenocarcinoma group was found (P = 0.003). Thus, our findings imply that the GSTMI and CYP1A1 exon 7 polymorphisms may influence PAH-DNA adduct levels in target tissue from NSCLC patients, especially in the squamous cell carcinoma group. Moreover, individuals carrying the GSTM1(null)/CYP1A1-Ile/Val genotype might exhibit a greater predisposition to a peripheral type of lung cancer.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/genética , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP2D6/genética , Adutos de DNA/genética , Poluentes Ambientais/metabolismo , Glutationa Transferase/genética , Neoplasias Pulmonares/genética , Polimorfismo Genético/genética , Fumar/genética , Adulto , Idoso , Carcinoma Pulmonar de Células não Pequenas/enzimologia , Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP2D6/metabolismo , Adutos de DNA/metabolismo , Suscetibilidade a Doenças , Poluentes Ambientais/efeitos adversos , Feminino , Genótipo , Glutationa Transferase/metabolismo , Humanos , Neoplasias Pulmonares/enzimologia , Masculino , Pessoa de Meia-Idade , Polônia , Fumar/efeitos adversos , Fumar/metabolismoRESUMO
Genotoxic agents of environmental or dietary origin may play a role in breast cancer initiation. The ability of extracts of human milk to cause mutations in S. typhimurium TA1538 and YG1019 and to induce micronuclei and DNA strand breaks in MCL-5 cells was investigated. Twenty samples from different donors were analysed and of these, 6 were adjudged to produce positive mutagenic response in one or both bacterial strains. The same samples also induced significant micronucleus formation in MCL-5 cells. In the comet assay, 13/20 samples caused DNA strand breaks in MCL-5 cells. Viable exfoliated breast cells were recovered from fresh milk samples and the ability of milk extracts to cause DNA damage in these cells was demonstrated. The results show that human milk can contain components capable of causing genotoxic damage in test systems and in human breast cells, events that may be significant in the initiation of breast cancer
Assuntos
Dano ao DNA , Células Epiteliais/efeitos dos fármacos , Leite Humano/citologia , Mutagênicos/toxicidade , Mama/citologia , Neoplasias da Mama/etiologia , Neoplasias da Mama/genética , Linhagem Celular , Células Cultivadas , DNA/efeitos dos fármacos , DNA/genética , Células Epiteliais/ultraestrutura , Feminino , Humanos , Lactação , Testes para Micronúcleos , Leite Humano/química , Testes de Mutagenicidade , Mutagênicos/isolamento & purificação , Mutagênicos/farmacologia , Mutação , Projetos Piloto , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/genética , Fatores de TempoRESUMO
Heterocyclic aromatic amines (HAAs), formed during the cooking of foods, are known to induce tumours in rodent bioassays and may thus contribute to human cancer risk. We tested six HAAs in a morphological transformation assay and in three in vitro genotoxicity assays. The morphological transforming abilities of HAAs were tested, in the presence of rat-liver S9, in the C3H/M2 fibroblast cell line. Concentration levels of 50 microM 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (8-MeIQx), 100 microM 2-amino-3,4,8-trimethylimidazo-[4,5-f]quinoxaline (4,8-DiMeIQx), 50 microM 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 100 microM 2-amino-9H-pyrido[2,3-b]indole (AalphaC), 100 microM 2-amino-3-methyl-9H-pyrido[2,3-b]indole (MeAalphaC) and 15 microM 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) induced maximum transformation potencies of 5.5, 6.6, 6.3, 5.2, 7.3 and 9.2 transformed foci per 10(4) surviving cells, respectively. Bacterial mutagenic activity was determined in the presence of rat-liver S9 using the Salmonella typhimurium reverse-mutation assay employing strain YG1019. Mutagenic potencies of 3800 revertants (revs)/ng with 8-MeIQx, 2900 revs/ng with 4,8-DiMeIQx, 3480 revs/ng with IQ, 1.6 revs/ng with AalphaC, 2.9 revs/ng with MeAalphaC and 5 revs/ng with PhIP were observed. Clastogenic activity in vitro was analysed by the micronucleus assay in metabolically competent MCL-5 cells. Dose-dependent induction of micronuclei was observed for all HAAs tested with 1-5.4% of cells containing micronuclei at 10 ng/ml. Micronucleus induction was in the order 4,8-DiMeIQx > 8-MeIQx > IQ > MeAalphaC > PhIP > AalphaC. DNA strand-breaking activity in MCL-5 cells was measured by the alkaline single cell-gel (comet) assay. The lowest effect doses for significant increases (P < or = 0.0007, Mann-Whitney test) in comet tail length (microm) were 45.5 microg/ml (200 microM) for PhIP, 90.9 microg/ml (410-510 microM) for 4,8-DiMeIQx, IQ, MeAalphaC and AalphaC, and 454.5 microg/ml (2130 microM) for 8-MeIQx. It is not yet clear which of these assays most accurately reflects the genotoxic potential to humans of compounds of this class of environmental carcinogens.
Assuntos
Carcinógenos Ambientais/toxicidade , Transformação Celular Neoplásica/induzido quimicamente , Dano ao DNA , Imidazóis/toxicidade , Quinoxalinas/toxicidade , Animais , Biotransformação , Carbolinas/toxicidade , Sistema Enzimático do Citocromo P-450/metabolismo , Relação Dose-Resposta a Droga , Fibroblastos/efeitos dos fármacos , Alimentos , Temperatura Alta , Humanos , Linfócitos/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C3H , Testes para Micronúcleos , Microssomos Hepáticos/metabolismo , Testes de Mutagenicidade , Mutagênicos/toxicidade , Quinolinas/toxicidade , Ratos , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/genética , Relação Estrutura-AtividadeRESUMO
Genotoxic agents of environmental or dietary origin may play a role in breast cancer initiation. The ability of extracts of human milk to cause mutations in S. typhimurium TA1538 and YG1019 and to induce micronuclei and DNA strand breaks in MCL-5 cells was investigated. Twenty samples from different donors were analysed and of these, 6 were adjudged to produce a positive mutagenic response in one or both bacterial strains. The same samples also induced significant micronucleus formation in MCL-5 cells. In the comet assay, 13/20 samples caused DNA strand breaks in MCL-5 cells. Viable exfoliated breast cells were recovered from fresh milk samples and the ability of milk extracts to cause DNA damage in these cells was demonstrated. The results show that human milk can contain components capable of causing genotoxic damage in test systems and in human breast cells, events that may be significant in the initiation of breast cancer.
Assuntos
Dano ao DNA , Células Epiteliais/efeitos dos fármacos , Leite Humano/citologia , Mutagênicos/toxicidade , Mama/citologia , Neoplasias da Mama/etiologia , Neoplasias da Mama/genética , Células Cultivadas , Citarabina , Relação Dose-Resposta a Droga , Células Epiteliais/ultraestrutura , Feminino , Humanos , Hidroxiureia , Testes para Micronúcleos , Leite Humano/química , Testes de Mutagenicidade , Mutagênicos/isolamento & purificação , Mutagênicos/farmacologia , Projetos Piloto , Proteína S9 Ribossômica , Proteínas Ribossômicas , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/genética , Fatores de TempoRESUMO
We investigated changes across the adult life span of the fingertip forces used to grip and lift objects and their possible causes. Grip force, relative safety margin (grip force exceeding the minimum to avoid slip, as a fraction of slip force), and skin slipperiness increased beginning at age 50 years. Skin slipperiness explained relative safety margin increases until age 60 years. Hence, after age 60 years, additional factors must elevate grip force. We argue that one factor is impaired cutaneous afferent encoding of skin-object frictional properties on the basis of three findings. First, only subjects 60 years and older increased their relative safety margins when the friction of the gripped surfaces was varied randomly versus experiments that varied only object weight. Skin slipperiness did not account for this behavior. Second, these older subjects scaled the initial portion of their force trajectories for the slippery surface during experiments when friction was varied. Third, their grip force adjustments to new surfaces were delayed approximately 100 msec as compared with young subjects. Previous research has demonstrated that friction is signaled locally by fast-adapting afferents (FA I afferents), which decrease in number during old age. By contrast, adjustments triggered by object set-down, an event encoded by FA II afferents throughout the hand and wrist, were not delayed in our old subjects. Other findings included that anticipatory control of fingertip forces using memory of object weight was unimpaired in old age. Finally, old and young adults modulated their fingertip forces with equal smoothness and with similar relative intertrial variability.
Assuntos
Envelhecimento/fisiologia , Dedos , Força da Mão , Remoção , Adulto , Idoso , Idoso de 80 Anos ou mais , Análise de Variância , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Tato/fisiologiaRESUMO
Mammary lipid may act as a reservoir for genotoxins. Mammary lipid extracts (MLEs), obtained from eight UK women (21-41 years) undergoing reduction mammoplasty, were examined for their abilities to morphologically transform C3H/M2 mouse fibroblasts. Resultant transformation rates were 0.27, 0.33, 0.07, 0.29, 0.21, 0.00, 0.07, and 0.13 transformed foci/treated dish, respectively. Although the lipid-extraction procedure used was originally designed to extract heterocyclic aromatic amines (HAAs), liquid chromatography/mass spectroscopy (LC/MS) with selective ion monitoring has failed to detect HAAs in any of the lipid extracts so far examined. Genotoxicities were also assessed in S. typhimurium TA98 and in metabolically competent human (MCL-5) cells by the micronucleus and by the alkaline single-cell gel ("comet") assays. The MLEs induced bacterial mutagenicity rates ranging from 0 to 498 revertants/plate/g-lipid equivalent and micronucleus-formation rates from 0 to 20 micronuclei/500 binucleate cells/g-lipid. Median comet tail lengths (induced with MLEs of 8.0 g-lipid equivalent) ranged from 6.0 to 74.0 micrometer. The results demonstrate the presence of as-yet-unidentified transforming agents in mammary lipid.
Assuntos
Mama/química , Fibroblastos/citologia , Lipídeos/isolamento & purificação , Lipídeos/farmacologia , Adulto , Animais , Linhagem Celular Transformada , Cruzamentos Genéticos , Dano ao DNA/genética , Feminino , Fibroblastos/efeitos dos fármacos , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Linfócitos/citologia , Linfócitos/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C3H , Testes para Micronúcleos , Mutagênese/efeitos dos fármacos , Mutagênese/genética , Testes de Mutagenicidade , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/genéticaRESUMO
This experiment addressed the often-posed theory that age-related declines in manual dexterity result from diminished tactile function. We measured the time 'young' subjects (n=33; mean=45 years) and 'old' subjects (n=33; mean=74 years) needed to grip (thumb and index finger), lift, and transport a small metal sphere when vision was permitted and when blindfolded. Subjects began each trial by reaching for the sphere and were instructed to complete the entire task quickly. In the absence of visual information, placement of the finger and thumb for a secure grip and lift cannot be performed efficiently without tactile information. If age-related tactile changes are functionally significant for this task, then without visual information the 'old' group should show a disproportionate increase in the duration of the grip and lift phase of the task compared to the 'young' group. Perceptual thresholds for tactile pressure stimuli (Semmes-Weinstein filaments) confirmed well-known age-related changes. Age and vision effects were manifest mainly during the grip-lift phase (time from object contact to lift-off from its support surface), with the expected finding that the 'old' group required more time than 'young' group, regardless of visual condition. The main finding was that the 'grip-lift' duration in the 'no-vision' condition was about twice the duration observed in the 'vision' condition for both age groups (ratios of 2.1 and 2.3 for 'young' and 'old', respectively). This similar relative slowing for the two groups fails to support the hypothesis that old adults' ability to grip and lift the object was limited by changes in the availability or use of tactile information.