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Cisplatin-DNA adducts in rat spermatozoa: detection and consequences
Book of Abstmcts - EUROTOX 94 ffesuffsand discussiax We did not find clinical sign related to diifunction of respiratorysystem. Different types of macroscopic pulmonary lesions could be found confirmed by histopathological examinations. Conclusion:The various nonneoplastic pulmonary lesions occurred in W&TAR rats in the subchronic tow stud. ies are supposed to be miscellaneous origin. Reference: Monographs on Pathology of LaboratoryAnimals Sponsored by the international Life Sciences Institute “Respiratory system” (T.C. Jones. U. Mohr Ft.0. Hunt) (1995). Key words: WISTAFI rat; nonneoplastic pulmonary lesions
Analysisof MutationsInducedby 2-Aml~l-Methyt~Phenyll~~[4,~]~l~ In Human LymphobtastoldCells 9. tfolzhauser. J. Horlbeck. f?Leong Morgen~al~ Swikerfand
Cerrfrede remerge
(PhIP)
Nest& Nestec S4, ~~~h~~s~nc,
The formation and presence of heterocyclic amines (HAA) in cooked meat and fiih is well documented [I], Of these compounds, PhlP has been found to be present in high content. Though PhlP has been found to be a relatively weak mutagen compared to other HAA including MelQ and MeQ in the Ames Assay, resutts from ~rnrna~~n show PhlP to be more active. In this abstract, we show that PhlP is mutagenic to the human lymphobfastoidceil line, TK6. In addition, we have analysed the mutations that are induced by this compound. The mutations identified in all the mutants characterised from the PhlP treated cultures are found to be on GC base pairs. In addition, most of the m~ations were GC-TA transversions.Assuming that all the adducts are on Gs. 90% of the mutations were found to result from the non-transcribed strand. Nearest neighbour analysis of the identified mutations show that there is a tendency for the presence of a purine on the 5’ and 3’ neighbor of the mutated G. [I] JS felton and MG Knize in Handbook of Experimental Pharmacology,Vol9411 pp. 471-502. Key words: food mutagens; human cells; mutational specificity
FactorsAffectingMortatityIn liunorigenlcttyStudlesConductedon Gang-Housed Sp~g~Da~ey Rats W.N. Hooks, C. Gopinath. D.F. Newton, R.J. Vsrling. ffun*ingdon Research Centre ftd,, Huntingdon, Cambridgeshire, UK. A review of the mortality in 70 Sp~gu~Dawley rat tumo~gen~i~ studies, performed in our laboratories, has shown that changing from a high protein breeding diet to a lower protein maintenance diet resulted in lower mortality in both dietary and oral gavage administered studies 111.Subsequently, it was decided to investigate the mortalities in greater detail in order to identify the underlying factors. Over the first 52 weeks of both dietary and oral gavage studies, although animals fed maintenance diet showed higher food consumption, they had lower bodyweight gain. This was considered to reflect the lower overall protein intake in comparison to animals fed breeding diet. Further review of the bodyweight gains in maintenance diet studies has also revealed that the mean gain of decedent animals was greater than that of animals survivingto termination. Analysisof pathological data from representative studies has shown that in maintenance diet studies there was a lower incidence of islet cell tumours of the pancreas. and in the females only. a lower incidence of tumour bearing rats, liver tumours and in mammal carcinoma. However, the principal pathological factor associated with the k>wer mortality seen for animals fed maintenance diet was a reduction in the incidence of progressiveglomerulonephrosis,which was probablyassociated with the lower protein intake. [l f Hooks, W N et al, The Toxicologist.13,419 (1993). Key words: mortality; Sprague-Dawley rats; tumorigenicity
Ci~~tt~DNA AdductsIn Rat SpermatozoaiDetscttonand Consequances S.B. Hooser. A.M.J. Fichtinger-Schepman,R.A. Baan, A.E. Smits-van Prooije. D.H. Waalkens-6erendsen, N.J. Snoe& Division of Toxico/om TN0 Nutrition and rood Reseen%rInstitute, Zeist llre Netherlands Ciiplatin IcisPt) is a widely used antineoplastic drug, utilii in the treatment of testicular, ovarian and head and neck cancers. It is thought to exert its antitumour effect through binding to DNA and forming &P&DNA adducts. These studies were done to determine if DNA adducts can beformed in rat spermatozoa following cisPt treatment in vitm or in v&o: cisPt adducts were detected by means of an ELISA method. In spermatozoa isolated from the
38
Book of Abshucts - EVROTOX ‘94
cauda epididymes of adult, male Wistar rats, 3 types of bifunctional cisPt-DNA adducts could be detected after incubation with 10 or 50 pg cisplatinlml for 15 minutes. Total DNA adduct levels were 3 and 17 fmofeslflg DNA, respectively. lmmunofluorescence analysis showed that all spermatozoa contain DNA adducts following in vitro exposure. lntraperitonealtreatment of adult, male Wistar rats with 10 mg cisPt/kg b.w., induced detectable numbers of cisPt-DNA adducts in epididymal spermatozoa at 6 hrs after dosing (0.6 fmollpg DNA). Adduct levels increased to 0.9 fmof/lg DNA at 24 hrs and gradually declined to 0.3 pg DNA at 7 days after treatment. The inguence of these cisPt adducts on reproductive performance and fetal development were studied by mating male rats treated with 10 mg cisPt/kg. i.p. with untreated females. Reproduction parameters appeared normal and upon caesarian section at day 21 of gestation no abnormalities were observed. Key words: cisplatin; DNA adducts; rat sperm; reproduction
Environmental and Occupational-Associated Hematological Changes V Hristeva-Mircheva. National Centre of /-&giene, Medical Ecology Examinations of the peripheral blood cells are carried out on 105 workers of the Phenol Installation at a Petrol Chemical Plant (PCHP). All workers live in four villages and a town situated around the PCPH. Hematological indices of iO0 subjects from the population of these places and 50 subjects as a “clear control group” were investigated too. The workers are occupationalexpose to a phenol, acetone and isopropylbenzene. The atmospheric air around the PCHP is periodicallycontaminated with H$S. SO2. N02. phenol and styrene. The performed study of peripheral blood elements resulted in the following findings: an anemia is met statistically srgnifirant more often in the workers group than in the subjects of the “clear control group”; in 13.3% of the workers an eosinophylia is presented; in 10.4% of the workers and 6% of the investigated urban population a leucopenia or tendency to such one is established. Key words: blood cells: workers; population; phenol: isopropylbenzene
The Application of Human-vpe Diets in Rodent Feeding Studies for the Safety Evaluatlon of Novel Foods and Processes A. Huaaett, J.C. Tschantz.A. Donnet, M. Marchesini. I. Perrin. B. Schilter, G. Sunahara. H.-P Wurzner. Deparrment of Quality and SafetyAssurance, Nestec Ltd. Research Centre, Vers-chez-les-Blanc,Lausanne, Switlerland Traditionalfood safety evaluation procedures, includingOO-daysubchronic rodent toxicology studies, were designed for additives and other components consumed in small quantities and comprising an insignificant proportion of the diet. The development of novelfoods which may comprise a more substantial part of the diet, pose several problems with regard to their safety evaluation by conventionaltesting procedures. These may result from nutritional disturbances caused by addition of large quantities of test material to the basal rodent diet. Alternatively the significance of the effects of novel processing/treatment procedures(e.g. microwave cooking) on non-human diets may be questionable. In order to overcome some of these problems we have investigated the application of human-type diets, with interchangeable macroconstituents. adapted for the nutritional needs of rodents. A go-day subchronicfeeding study in rats was used to compare the effects of a human-type diet based on industrial dried and precooked ingredients (carrots. potatoes, green beans, couscous, lentils and granulated meat) with a conventional semi-synthetic rodent diet. Various toxicological parameters were examined including clinical chemistry, haematology. histology, immunological markers. and xenobiotic metabolising activities. Although the results showed some differences in certain of the parameters measured there were no indications of any adverse effects, thus demonstrating the feasibility of using human-type diets in rodent subchronicfeeding studies. In conclusion the use of modified basal diets in toxicological studies may provide a practical approach for the safety evaluation of novel foods. Key words: novel foods; novel processes; human-type diets; subchronic toxicity; rat
Mechanisms of Lung Diseases Development After Asbestos Exposure M. Hurbrinkod. A. KaiglovB.T Trnovec. Institute of Reventive and Clinical Medicine, Bratislava,Slovakia Lung macrophages exposed to asbestos release a bioactive oxygen radicals, which stimulate the inflammatory cells to the secretion of various mediators, cytokines and growth factors. Growth factors induce the proliferation of fibroblasts leading to increased collagen deposition, IL-l - release from pulmonary macrophages associated with decreased production of suppressor T-cells can provoke the B-lymphocyte hyperfunction - increased levels of immunoglobulines 111.We investigated some of the parameters of humaral immunity in workers exposed to asbestos
Analysisof MutationsInducedby 2-Aml~l-Methyt~Phenyll~~[4,~]~l~ In Human LymphobtastoldCells 9. tfolzhauser. J. Horlbeck. f?Leong Morgen~al~ Swikerfand
Cerrfrede remerge
(PhIP)
Nest& Nestec S4, ~~~h~~s~nc,
The formation and presence of heterocyclic amines (HAA) in cooked meat and fiih is well documented [I], Of these compounds, PhlP has been found to be present in high content. Though PhlP has been found to be a relatively weak mutagen compared to other HAA including MelQ and MeQ in the Ames Assay, resutts from ~rnrna~~n show PhlP to be more active. In this abstract, we show that PhlP is mutagenic to the human lymphobfastoidceil line, TK6. In addition, we have analysed the mutations that are induced by this compound. The mutations identified in all the mutants characterised from the PhlP treated cultures are found to be on GC base pairs. In addition, most of the m~ations were GC-TA transversions.Assuming that all the adducts are on Gs. 90% of the mutations were found to result from the non-transcribed strand. Nearest neighbour analysis of the identified mutations show that there is a tendency for the presence of a purine on the 5’ and 3’ neighbor of the mutated G. [I] JS felton and MG Knize in Handbook of Experimental Pharmacology,Vol9411 pp. 471-502. Key words: food mutagens; human cells; mutational specificity
FactorsAffectingMortatityIn liunorigenlcttyStudlesConductedon Gang-Housed Sp~g~Da~ey Rats W.N. Hooks, C. Gopinath. D.F. Newton, R.J. Vsrling. ffun*ingdon Research Centre ftd,, Huntingdon, Cambridgeshire, UK. A review of the mortality in 70 Sp~gu~Dawley rat tumo~gen~i~ studies, performed in our laboratories, has shown that changing from a high protein breeding diet to a lower protein maintenance diet resulted in lower mortality in both dietary and oral gavage administered studies 111.Subsequently, it was decided to investigate the mortalities in greater detail in order to identify the underlying factors. Over the first 52 weeks of both dietary and oral gavage studies, although animals fed maintenance diet showed higher food consumption, they had lower bodyweight gain. This was considered to reflect the lower overall protein intake in comparison to animals fed breeding diet. Further review of the bodyweight gains in maintenance diet studies has also revealed that the mean gain of decedent animals was greater than that of animals survivingto termination. Analysisof pathological data from representative studies has shown that in maintenance diet studies there was a lower incidence of islet cell tumours of the pancreas. and in the females only. a lower incidence of tumour bearing rats, liver tumours and in mammal carcinoma. However, the principal pathological factor associated with the k>wer mortality seen for animals fed maintenance diet was a reduction in the incidence of progressiveglomerulonephrosis,which was probablyassociated with the lower protein intake. [l f Hooks, W N et al, The Toxicologist.13,419 (1993). Key words: mortality; Sprague-Dawley rats; tumorigenicity
Ci~~tt~DNA AdductsIn Rat SpermatozoaiDetscttonand Consequances S.B. Hooser. A.M.J. Fichtinger-Schepman,R.A. Baan, A.E. Smits-van Prooije. D.H. Waalkens-6erendsen, N.J. Snoe& Division of Toxico/om TN0 Nutrition and rood Reseen%rInstitute, Zeist llre Netherlands Ciiplatin IcisPt) is a widely used antineoplastic drug, utilii in the treatment of testicular, ovarian and head and neck cancers. It is thought to exert its antitumour effect through binding to DNA and forming &P&DNA adducts. These studies were done to determine if DNA adducts can beformed in rat spermatozoa following cisPt treatment in vitm or in v&o: cisPt adducts were detected by means of an ELISA method. In spermatozoa isolated from the
38
Book of Abshucts - EVROTOX ‘94
cauda epididymes of adult, male Wistar rats, 3 types of bifunctional cisPt-DNA adducts could be detected after incubation with 10 or 50 pg cisplatinlml for 15 minutes. Total DNA adduct levels were 3 and 17 fmofeslflg DNA, respectively. lmmunofluorescence analysis showed that all spermatozoa contain DNA adducts following in vitro exposure. lntraperitonealtreatment of adult, male Wistar rats with 10 mg cisPt/kg b.w., induced detectable numbers of cisPt-DNA adducts in epididymal spermatozoa at 6 hrs after dosing (0.6 fmollpg DNA). Adduct levels increased to 0.9 fmof/lg DNA at 24 hrs and gradually declined to 0.3 pg DNA at 7 days after treatment. The inguence of these cisPt adducts on reproductive performance and fetal development were studied by mating male rats treated with 10 mg cisPt/kg. i.p. with untreated females. Reproduction parameters appeared normal and upon caesarian section at day 21 of gestation no abnormalities were observed. Key words: cisplatin; DNA adducts; rat sperm; reproduction
Environmental and Occupational-Associated Hematological Changes V Hristeva-Mircheva. National Centre of /-&giene, Medical Ecology Examinations of the peripheral blood cells are carried out on 105 workers of the Phenol Installation at a Petrol Chemical Plant (PCHP). All workers live in four villages and a town situated around the PCPH. Hematological indices of iO0 subjects from the population of these places and 50 subjects as a “clear control group” were investigated too. The workers are occupationalexpose to a phenol, acetone and isopropylbenzene. The atmospheric air around the PCHP is periodicallycontaminated with H$S. SO2. N02. phenol and styrene. The performed study of peripheral blood elements resulted in the following findings: an anemia is met statistically srgnifirant more often in the workers group than in the subjects of the “clear control group”; in 13.3% of the workers an eosinophylia is presented; in 10.4% of the workers and 6% of the investigated urban population a leucopenia or tendency to such one is established. Key words: blood cells: workers; population; phenol: isopropylbenzene
The Application of Human-vpe Diets in Rodent Feeding Studies for the Safety Evaluatlon of Novel Foods and Processes A. Huaaett, J.C. Tschantz.A. Donnet, M. Marchesini. I. Perrin. B. Schilter, G. Sunahara. H.-P Wurzner. Deparrment of Quality and SafetyAssurance, Nestec Ltd. Research Centre, Vers-chez-les-Blanc,Lausanne, Switlerland Traditionalfood safety evaluation procedures, includingOO-daysubchronic rodent toxicology studies, were designed for additives and other components consumed in small quantities and comprising an insignificant proportion of the diet. The development of novelfoods which may comprise a more substantial part of the diet, pose several problems with regard to their safety evaluation by conventionaltesting procedures. These may result from nutritional disturbances caused by addition of large quantities of test material to the basal rodent diet. Alternatively the significance of the effects of novel processing/treatment procedures(e.g. microwave cooking) on non-human diets may be questionable. In order to overcome some of these problems we have investigated the application of human-type diets, with interchangeable macroconstituents. adapted for the nutritional needs of rodents. A go-day subchronicfeeding study in rats was used to compare the effects of a human-type diet based on industrial dried and precooked ingredients (carrots. potatoes, green beans, couscous, lentils and granulated meat) with a conventional semi-synthetic rodent diet. Various toxicological parameters were examined including clinical chemistry, haematology. histology, immunological markers. and xenobiotic metabolising activities. Although the results showed some differences in certain of the parameters measured there were no indications of any adverse effects, thus demonstrating the feasibility of using human-type diets in rodent subchronicfeeding studies. In conclusion the use of modified basal diets in toxicological studies may provide a practical approach for the safety evaluation of novel foods. Key words: novel foods; novel processes; human-type diets; subchronic toxicity; rat
Mechanisms of Lung Diseases Development After Asbestos Exposure M. Hurbrinkod. A. KaiglovB.T Trnovec. Institute of Reventive and Clinical Medicine, Bratislava,Slovakia Lung macrophages exposed to asbestos release a bioactive oxygen radicals, which stimulate the inflammatory cells to the secretion of various mediators, cytokines and growth factors. Growth factors induce the proliferation of fibroblasts leading to increased collagen deposition, IL-l - release from pulmonary macrophages associated with decreased production of suppressor T-cells can provoke the B-lymphocyte hyperfunction - increased levels of immunoglobulines 111.We investigated some of the parameters of humaral immunity in workers exposed to asbestos