^STUDY !CHEMICAL_NAME=C.I. Direct blue 218 !STUDY_CONCLUSION=Weakly Positive !DEPOSITOR_STUDY_NUMBER=376714_SCE !STRESSOR_TYPE=CHEMICAL !INSTITUTION=NTP !STUDY_TITLE=Cytogenetic study of C.I. Direct blue 218 in Chinese Hamster Ovary Cell Sister Chromatid Exchange Test !STUDY_DESIGN=Parallel !SUBJECT_TYPE=IN_VITRO !OLD_NTP_STUDY_TYPE=CHO_SCE !NTP_STUDY_TYPE=Genetic Toxicology - Mammalian Cell Cytogenetics !TIME_TYPE=Study !STUDY_VARIABLES=COMPOUND|MICROSOMAL_ACTIVATION_USED|DOSE|DOSE_UNIT|TRIAL_RESULT|TREATMENT_GROUP_TYPE|START_DATE ^GROUP #GROUP_NAME #COMPARATOR_NAME #IS_CONTROL_GROUP #SUPER_GROUP_NAME #TREATMENT_GROUP_TYPE #DEPOSITOR_NAME #ASSAY_TRIAL #COMPOUND #SPECIES_COMMON_NAME #CELL_LINE #STRAIN #MICROSOMAL_ACTIVATION_USED #MICROSOMAL_ACTIVATION_SPECIES #DOSE #DOSE_UNIT #TRIAL_RESULT #START_DATE #CONTROL_SUBSTANCE_NAME $GROUP_NAME $COMPARATOR_NAME $IS_CONTROL_GROUP $SUPER_GROUP_NAME $TREATMENT_GROUP_TYPE $DEPOSITOR_NAME $ASSAY_TRIAL $COMPOUND $SPECIES_COMMON_NAME $CELL_LINE $STRAIN $MICROSOMAL_ACTIVATION_USED $MICROSOMAL_ACTIVATION_SPECIES $DOSE $DOSE_UNIT $TRIAL_RESULT $START_DATE $CONTROL_SUBSTANCE_NAME 1 1 Hamster Chinese Hamster Ovary CHO-W-B1 No Activation N_A Negative 1982-10-10 1_1_Medium T 1 Vehicle Control 1 Medium Hamster Chinese Hamster Ovary CHO-W-B1 0 Medium 1_2_Negative (Not Specified) T 1 Vehicle Control 2 Negative (Not Specified) Hamster Chinese Hamster Ovary CHO-W-B1 0 Negative (Not Specified) 1_3_0.16_ug/mL_C.I. Direct blue 218 1_2_Negative (Not Specified) 1 Test Chemical 3 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 0.16 ug/mL 1_4_5.0_ug/mL_C.I. Direct blue 218 1_2_Negative (Not Specified) 1 Test Chemical 4 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 5.0 ug/mL 1_5_16.0_ug/mL_C.I. Direct blue 218 1_2_Negative (Not Specified) 1 Test Chemical 5 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 16.0 ug/mL 1_6_50.0_ug/mL_C.I. Direct blue 218 1_2_Negative (Not Specified) 1 Test Chemical 6 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 50.0 ug/mL 1_7_160.0_ug/mL_C.I. Direct blue 218 1_2_Negative (Not Specified) 1 Test Chemical 7 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 160.0 ug/mL 1_8_0.01_ug/mL_Mitomycin-C 1_2_Negative (Not Specified) T 1 Positive Control 8 Mitomycin-C Hamster Chinese Hamster Ovary CHO-W-B1 0.01 ug/mL Mitomycin-C 2 2 Hamster Chinese Hamster Ovary CHO-W-B1 No Activation N_A Weakly Positive 1982-10-20 2_1_Negative (Not Specified) T 2 Vehicle Control 1 Negative (Not Specified) Hamster Chinese Hamster Ovary CHO-W-B1 0 Negative (Not Specified) 2_2_Medium T 2 Vehicle Control 2 Medium Hamster Chinese Hamster Ovary CHO-W-B1 0 Medium 2_3_25.0_ug/mL_C.I. Direct blue 218 2_2_Medium 2 Test Chemical 3 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 25.0 ug/mL 2_4_50.0_ug/mL_C.I. Direct blue 218 2_2_Medium 2 Test Chemical 4 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 50.0 ug/mL 2_5_75.0_ug/mL_C.I. Direct blue 218 2_2_Medium 2 Test Chemical 5 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 75.0 ug/mL 2_6_100.0_ug/mL_C.I. Direct blue 218 2_2_Medium 2 Test Chemical 6 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 100.0 ug/mL 2_7_200.0_ug/mL_C.I. Direct blue 218 2_2_Medium 2 Test Chemical 7 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 200.0 ug/mL 2_8_300.0_ug/mL_C.I. Direct blue 218 2_2_Medium 2 Test Chemical 8 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 300.0 ug/mL 2_9_0.005_ug/mL_Mitomycin-C 2_2_Medium T 2 Positive Control 9 Mitomycin-C Hamster Chinese Hamster Ovary CHO-W-B1 0.005 ug/mL Mitomycin-C 2_10_0.01_ug/mL_Mitomycin-C 2_2_Medium T 2 Positive Control 10 Mitomycin-C Hamster Chinese Hamster Ovary CHO-W-B1 0.01 ug/mL Mitomycin-C 1_S9 1_S9 Hamster Chinese Hamster Ovary CHO-W-B1 Induced Rat Liver S9 RAT Negative 1982-10-10 1_S9_1_Medium T 1_S9 Vehicle Control 1 Medium Hamster Chinese Hamster Ovary CHO-W-B1 0 Medium 1_S9_2_Negative (Not Specified) T 1_S9 Vehicle Control 2 Negative (Not Specified) Hamster Chinese Hamster Ovary CHO-W-B1 0 Negative (Not Specified) 1_S9_3_1.6_ug/mL_C.I. Direct blue 218 1_S9_2_Negative (Not Specified) 1_S9 Test Chemical 3 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 1.6 ug/mL 1_S9_4_5.0_ug/mL_C.I. Direct blue 218 1_S9_2_Negative (Not Specified) 1_S9 Test Chemical 4 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 5.0 ug/mL 1_S9_5_16.0_ug/mL_C.I. Direct blue 218 1_S9_2_Negative (Not Specified) 1_S9 Test Chemical 5 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 16.0 ug/mL 1_S9_6_50.0_ug/mL_C.I. Direct blue 218 1_S9_2_Negative (Not Specified) 1_S9 Test Chemical 6 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 50.0 ug/mL 1_S9_7_160.0_ug/mL_C.I. Direct blue 218 1_S9_2_Negative (Not Specified) 1_S9 Test Chemical 7 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 160.0 ug/mL 1_S9_8_1.5_ug/mL_Cyclophosphamide 1_S9_2_Negative (Not Specified) T 1_S9 Positive Control 8 Cyclophosphamide Hamster Chinese Hamster Ovary CHO-W-B1 1.5 ug/mL Cyclophosphamide 2_S9 2_S9 Hamster Chinese Hamster Ovary CHO-W-B1 Induced Rat Liver S9 RAT Questionable 1982-10-29 2_S9_1_Medium T 2_S9 Vehicle Control 1 Medium Hamster Chinese Hamster Ovary CHO-W-B1 0 Medium 2_S9_2_Negative (Not Specified) T 2_S9 Vehicle Control 2 Negative (Not Specified) Hamster Chinese Hamster Ovary CHO-W-B1 0 Negative (Not Specified) 2_S9_3_25.0_ug/mL_C.I. Direct blue 218 2_S9_2_Negative (Not Specified) 2_S9 Test Chemical 3 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 25.0 ug/mL 2_S9_4_50.0_ug/mL_C.I. Direct blue 218 2_S9_2_Negative (Not Specified) 2_S9 Test Chemical 4 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 50.0 ug/mL 2_S9_5_100.0_ug/mL_C.I. Direct blue 218 2_S9_2_Negative (Not Specified) 2_S9 Test Chemical 5 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 100.0 ug/mL 2_S9_6_150.0_ug/mL_C.I. Direct blue 218 2_S9_2_Negative (Not Specified) 2_S9 Test Chemical 6 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 150.0 ug/mL 2_S9_7_200.0_ug/mL_C.I. Direct blue 218 2_S9_2_Negative (Not Specified) 2_S9 Test Chemical 7 C.I. Direct blue 218 Hamster Chinese Hamster Ovary CHO-W-B1 200.0 ug/mL 2_S9_8_1.5_ug/mL_Cyclophosphamide 2_S9_2_Negative (Not Specified) T 2_S9 Positive Control 8 Cyclophosphamide Hamster Chinese Hamster Ovary CHO-W-B1 1.5 ug/mL Cyclophosphamide ^STRESSOR_PROTOCOL=CHEM_STRESSOR_PROTOCOL #PROTOCOL_DISPLAY_NAME #PROTOCOL_DESC #STRESSOR_NAME #DOSE_PER_ADMIN #DOSE_PER_ADMIN_UNIT #VEHICLE_NAME #VEHICLE_CONCENTRATION #VEHICLE_CONC_UNIT #IS_POSITIVE_CONTROL #IS_NEGATIVE_CONTROL #IS_VEHICLE_CONTROL #AGENT_SUPPLIER #AGENT_SUPPLIER_LOCATION $PROTOCOL_DISPLAY_NAME $PROTOCOL_DESC $STRESSOR_NAME $DOSE_PER_ADMIN $DOSE_PER_ADMIN_UNIT $VEHICLE_NAME $VEHICLE_CONCENTRATION $VEHICLE_CONC_UNIT $IS_POSITIVE_CONTROL $IS_NEGATIVE_CONTROL $IS_VEHICLE_CONTROL $AGENT_SUPPLIER $AGENT_SUPPLIER_LOCATION C.I. Direct blue 218_16.0_ug/mL C.I. Direct blue 218 16.0 ug/mL DMSO 10 ug/ml Radian Corporation Austin, TX Cyclophosphamide_1.5_ug/mL Cyclophosphamide 1.5 ug/mL DMSO 10 ug/ml T Radian Corporation Austin, TX DMSO No stressor 0 DMSO 10 ug/ml T Mitomycin-C_0.01_ug/mL Mitomycin-C 0.01 ug/mL DMSO 10 ug/ml T Radian Corporation Austin, TX C.I. Direct blue 218_1.6_ug/mL C.I. Direct blue 218 1.6 ug/mL DMSO 10 ug/ml Radian Corporation Austin, TX C.I. Direct blue 218_160.0_ug/mL C.I. Direct blue 218 160.0 ug/mL DMSO 10 ug/ml Radian Corporation Austin, TX C.I. Direct blue 218_25.0_ug/mL C.I. Direct blue 218 25.0 ug/mL DMSO 10 ug/ml Radian Corporation Austin, TX C.I. Direct blue 218_100.0_ug/mL C.I. Direct blue 218 100.0 ug/mL DMSO 10 ug/ml Radian Corporation Austin, TX C.I. Direct blue 218_5.0_ug/mL C.I. Direct blue 218 5.0 ug/mL DMSO 10 ug/ml Radian Corporation Austin, TX C.I. Direct blue 218_200.0_ug/mL C.I. Direct blue 218 200.0 ug/mL DMSO 10 ug/ml Radian Corporation Austin, TX Mitomycin-C_0.005_ug/mL Mitomycin-C 0.005 ug/mL DMSO 10 ug/ml T Radian Corporation Austin, TX C.I. Direct blue 218_0.16_ug/mL C.I. Direct blue 218 0.16 ug/mL DMSO 10 ug/ml Radian Corporation Austin, TX C.I. Direct blue 218_150.0_ug/mL C.I. Direct blue 218 150.0 ug/mL DMSO 10 ug/ml Radian Corporation Austin, TX C.I. Direct blue 218_300.0_ug/mL C.I. Direct blue 218 300.0 ug/mL DMSO 10 ug/ml Radian Corporation Austin, TX C.I. Direct blue 218_50.0_ug/mL C.I. Direct blue 218 50.0 ug/mL DMSO 10 ug/ml Radian Corporation Austin, TX C.I. Direct blue 218_75.0_ug/mL C.I. Direct blue 218 75.0 ug/mL DMSO 10 ug/ml Radian Corporation Austin, TX ^CARE=CULTURE_PROTOCOL #PROTOCOL_DISPLAY_NAME #PROTOCOL_DESC #LIGHT #MEDIUM #SUPPLEMENTS #SERUM_PERCENTAGE #SERUM_TYPE #ACTIVATION #SPECIES #GAS_PHASE_PERCENT_CO2 #CULTURE_TEMPERATURE_MAX #CULTURE_TEMPERATURE_UNIT #IS_STOP $PROTOCOL_DISPLAY_NAME $PROTOCOL_DESC $LIGHT $MEDIUM $SUPPLEMENTS $SERUM_PERCENTAGE $SERUM_TYPE $ACTIVATION $SPECIES $GAS_PHASE_PERCENT_CO2 $CULTURE_TEMPERATURE_MAX $CULTURE_TEMPERATURE_UNIT $IS_STOP SCE_CULTURE Assay without metabolic activation. One day after plating, medium replaced with medium containing with test or control chemicals. After 2 hr BrdUrd was added to 10 uM. Cells were cultured for about 24 hr, medium replaced with fresh medium containing 10 pM BrdUrd and colcemid (final concentration 0.1 pg/ml) was added, and incubation continued for 2 to 3 hr. The total incubation time with test chemical was thus about 26 hr, and total time with BrdUrd was also about 26 hr, beginning about 2 hr after addition of test chemical. Gold light McCoy's 5A medium 2 mM glutamine, LBI used 100 units/ ml penicillin and 100 ug/ml streptomycin, CU did not 10 Fetal calf 5 37 degree C F SCE_CULTURE_S9 Assay with metabolic activation. One day after plating, medium was replaced with S9 mix in medium without fetal calf serum. After 2 hr cells were washed at least twice with buffered saline, and culture medium containing 10% FCS and 10 uM BrdUrd was added. Cells were incubated for 26 hr, with colcemid (0.1 pg/ml) present for the final 2 to 3 hr. Gold light McCoy's 5A medium 2 mM glutamine, LBI used 100 units/ ml penicillin and 100 ug/ml streptomycin, CU did not 10 Fetal calf Aroclor 1254-induced male Sprague-Dawley rat, thawed and mixed with NADPH generating system (final 2.4 mg/ml NADP (Sodium salt), 4.5 mg/ml isocitric acid (trisodium salt), S9 15 ul/ml) Rat 5 37 degree C F ADD_BRDU Gold light BrdU to 10 uM 5 37 degree C F ADD_COLCEMID Gold light Colcemid to 0.1 ug/ml 5 37 degree C F CULTURE_IN_FRESH_MEDIUM Gold light McCoy's 5A medium 2 mM glutamine, LBI used 100 units/ ml penicillin and 100 ug/ml streptomycin, CU did not 10 Fetal calf 5 37 degree C T ^PREPARATION=CULTURE_PREPARATION_PROTOCOL #PROTOCOL_DISPLAY_NAME #PROTOCOL_DESC $PROTOCOL_DISPLAY_NAME $PROTOCOL_DESC MITOTIC_SHAKE_OFF Cell harvest and fixation. Two to three hours after addition of colcemid, cells were collected by mitotic shake-off and treated for up to 3 min at room temperature with hypotonic KCI (75 mM). Cells were then washed twice with fixative (3: 1, methano1:glacial acetic acid, v/v), dropped onto slides, and air-dried. ^TIMELINE #TIME_IN_STUDY #TIME_IN_STUDY_UNIT #GROUP_NAME #PROTOCOL_DISPLAY_NAME $TIME_IN_STUDY $TIME_IN_STUDY_UNIT $GROUP_NAME $PROTOCOL_DISPLAY_NAME 0 hour 1_1_Medium SCE_CULTURE 2 hour 1_1_Medium ADD_BRDU 26.0 hour 1_1_Medium ADD_COLCEMID 28.0 hour 1_1_Medium MITOTIC_SHAKE_OFF 0 hour 1_2_Negative (Not Specified) SCE_CULTURE 2 hour 1_2_Negative (Not Specified) ADD_BRDU 26.0 hour 1_2_Negative (Not Specified) ADD_COLCEMID 28.0 hour 1_2_Negative (Not Specified) MITOTIC_SHAKE_OFF 0 hour 1_3_0.16_ug/mL_C.I. Direct blue 218 SCE_CULTURE 2 hour 1_3_0.16_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 1_3_0.16_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 1_3_0.16_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 1_4_5.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE 2 hour 1_4_5.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 1_4_5.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 1_4_5.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 1_5_16.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE 2 hour 1_5_16.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 1_5_16.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 1_5_16.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 1_6_50.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE 2 hour 1_6_50.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 1_6_50.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 1_6_50.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 1_7_160.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE 2 hour 1_7_160.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 1_7_160.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 1_7_160.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 1_8_0.01_ug/mL_Mitomycin-C SCE_CULTURE 2 hour 1_8_0.01_ug/mL_Mitomycin-C ADD_BRDU 26.0 hour 1_8_0.01_ug/mL_Mitomycin-C ADD_COLCEMID 28.0 hour 1_8_0.01_ug/mL_Mitomycin-C MITOTIC_SHAKE_OFF 0 hour 2_1_Negative (Not Specified) SCE_CULTURE 2 hour 2_1_Negative (Not Specified) ADD_BRDU 26.0 hour 2_1_Negative (Not Specified) ADD_COLCEMID 28.0 hour 2_1_Negative (Not Specified) MITOTIC_SHAKE_OFF 0 hour 2_2_Medium SCE_CULTURE 2 hour 2_2_Medium ADD_BRDU 26.0 hour 2_2_Medium ADD_COLCEMID 28.0 hour 2_2_Medium MITOTIC_SHAKE_OFF 0 hour 2_3_25.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE 2 hour 2_3_25.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 2_3_25.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 2_3_25.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 2_4_50.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE 2 hour 2_4_50.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 2_4_50.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 2_4_50.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 2_5_75.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE 2 hour 2_5_75.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 2_5_75.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 2_5_75.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 2_6_100.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE 2 hour 2_6_100.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 2_6_100.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 2_6_100.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 2_7_200.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE 2 hour 2_7_200.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 2_7_200.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 2_7_200.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 2_8_300.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE 2 hour 2_8_300.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 0.0 hour 2_8_300.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 2.0 hour 2_8_300.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 2_9_0.005_ug/mL_Mitomycin-C SCE_CULTURE 2 hour 2_9_0.005_ug/mL_Mitomycin-C ADD_BRDU 26.0 hour 2_9_0.005_ug/mL_Mitomycin-C ADD_COLCEMID 28.0 hour 2_9_0.005_ug/mL_Mitomycin-C MITOTIC_SHAKE_OFF 0 hour 2_10_0.01_ug/mL_Mitomycin-C SCE_CULTURE 2 hour 2_10_0.01_ug/mL_Mitomycin-C ADD_BRDU 26.0 hour 2_10_0.01_ug/mL_Mitomycin-C ADD_COLCEMID 28.0 hour 2_10_0.01_ug/mL_Mitomycin-C MITOTIC_SHAKE_OFF 0 hour 1_S9_1_Medium SCE_CULTURE_S9 2 hour 1_S9_1_Medium CULTURE_IN_FRESH_MEDIUM 2 hour 1_S9_1_Medium ADD_BRDU 26.0 hour 1_S9_1_Medium ADD_COLCEMID 28.0 hour 1_S9_1_Medium MITOTIC_SHAKE_OFF 0 hour 1_S9_2_Negative (Not Specified) SCE_CULTURE_S9 2 hour 1_S9_2_Negative (Not Specified) CULTURE_IN_FRESH_MEDIUM 2 hour 1_S9_2_Negative (Not Specified) ADD_BRDU 26.0 hour 1_S9_2_Negative (Not Specified) ADD_COLCEMID 28.0 hour 1_S9_2_Negative (Not Specified) MITOTIC_SHAKE_OFF 0 hour 1_S9_3_1.6_ug/mL_C.I. Direct blue 218 SCE_CULTURE_S9 2 hour 1_S9_3_1.6_ug/mL_C.I. Direct blue 218 CULTURE_IN_FRESH_MEDIUM 2 hour 1_S9_3_1.6_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 1_S9_3_1.6_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 1_S9_3_1.6_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 1_S9_4_5.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE_S9 2 hour 1_S9_4_5.0_ug/mL_C.I. Direct blue 218 CULTURE_IN_FRESH_MEDIUM 2 hour 1_S9_4_5.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 1_S9_4_5.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 1_S9_4_5.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 1_S9_5_16.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE_S9 2 hour 1_S9_5_16.0_ug/mL_C.I. Direct blue 218 CULTURE_IN_FRESH_MEDIUM 2 hour 1_S9_5_16.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 1_S9_5_16.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 1_S9_5_16.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 1_S9_6_50.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE_S9 2 hour 1_S9_6_50.0_ug/mL_C.I. Direct blue 218 CULTURE_IN_FRESH_MEDIUM 2 hour 1_S9_6_50.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 1_S9_6_50.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 1_S9_6_50.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 1_S9_7_160.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE_S9 2 hour 1_S9_7_160.0_ug/mL_C.I. Direct blue 218 CULTURE_IN_FRESH_MEDIUM 2 hour 1_S9_7_160.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 1_S9_7_160.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 1_S9_7_160.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 1_S9_8_1.5_ug/mL_Cyclophosphamide SCE_CULTURE_S9 2 hour 1_S9_8_1.5_ug/mL_Cyclophosphamide CULTURE_IN_FRESH_MEDIUM 2 hour 1_S9_8_1.5_ug/mL_Cyclophosphamide ADD_BRDU 26.0 hour 1_S9_8_1.5_ug/mL_Cyclophosphamide ADD_COLCEMID 28.0 hour 1_S9_8_1.5_ug/mL_Cyclophosphamide MITOTIC_SHAKE_OFF 0 hour 2_S9_1_Medium SCE_CULTURE_S9 2 hour 2_S9_1_Medium CULTURE_IN_FRESH_MEDIUM 2 hour 2_S9_1_Medium ADD_BRDU 26.0 hour 2_S9_1_Medium ADD_COLCEMID 28.0 hour 2_S9_1_Medium MITOTIC_SHAKE_OFF 0 hour 2_S9_2_Negative (Not Specified) SCE_CULTURE_S9 2 hour 2_S9_2_Negative (Not Specified) CULTURE_IN_FRESH_MEDIUM 2 hour 2_S9_2_Negative (Not Specified) ADD_BRDU 26.0 hour 2_S9_2_Negative (Not Specified) ADD_COLCEMID 28.0 hour 2_S9_2_Negative (Not Specified) MITOTIC_SHAKE_OFF 0 hour 2_S9_3_25.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE_S9 2 hour 2_S9_3_25.0_ug/mL_C.I. Direct blue 218 CULTURE_IN_FRESH_MEDIUM 2 hour 2_S9_3_25.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 2_S9_3_25.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 2_S9_3_25.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 2_S9_4_50.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE_S9 2 hour 2_S9_4_50.0_ug/mL_C.I. Direct blue 218 CULTURE_IN_FRESH_MEDIUM 2 hour 2_S9_4_50.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 2_S9_4_50.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 2_S9_4_50.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 2_S9_5_100.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE_S9 2 hour 2_S9_5_100.0_ug/mL_C.I. Direct blue 218 CULTURE_IN_FRESH_MEDIUM 2 hour 2_S9_5_100.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 2_S9_5_100.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 2_S9_5_100.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 2_S9_6_150.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE_S9 2 hour 2_S9_6_150.0_ug/mL_C.I. Direct blue 218 CULTURE_IN_FRESH_MEDIUM 2 hour 2_S9_6_150.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 2_S9_6_150.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 2_S9_6_150.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 2_S9_7_200.0_ug/mL_C.I. Direct blue 218 SCE_CULTURE_S9 2 hour 2_S9_7_200.0_ug/mL_C.I. Direct blue 218 CULTURE_IN_FRESH_MEDIUM 2 hour 2_S9_7_200.0_ug/mL_C.I. Direct blue 218 ADD_BRDU 26.0 hour 2_S9_7_200.0_ug/mL_C.I. Direct blue 218 ADD_COLCEMID 28.0 hour 2_S9_7_200.0_ug/mL_C.I. Direct blue 218 MITOTIC_SHAKE_OFF 0 hour 2_S9_8_1.5_ug/mL_Cyclophosphamide SCE_CULTURE_S9 2 hour 2_S9_8_1.5_ug/mL_Cyclophosphamide CULTURE_IN_FRESH_MEDIUM 2 hour 2_S9_8_1.5_ug/mL_Cyclophosphamide ADD_BRDU 26.0 hour 2_S9_8_1.5_ug/mL_Cyclophosphamide ADD_COLCEMID 28.0 hour 2_S9_8_1.5_ug/mL_Cyclophosphamide MITOTIC_SHAKE_OFF 0 hour 2_6_100.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_100.0_ug/mL 0 hour 1_5_16.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_16.0_ug/mL 0 hour 2_9_0.005_ug/mL_Mitomycin-C Mitomycin-C_0.005_ug/mL 0 hour 2_5_75.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_75.0_ug/mL 0 hour 2_S9_4_50.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_50.0_ug/mL 0 hour 1_3_0.16_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_0.16_ug/mL 0 hour 1_2_Negative (Not Specified) DMSO 0 hour 1_S9_2_Negative (Not Specified) DMSO 0 hour 2_8_300.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_300.0_ug/mL 0 hour 2_S9_5_100.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_100.0_ug/mL 0 hour 1_4_5.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_5.0_ug/mL 0 hour 1_S9_3_1.6_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_1.6_ug/mL 0 hour 2_S9_1_Medium DMSO 0 hour 2_S9_3_25.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_25.0_ug/mL 0 hour 2_4_50.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_50.0_ug/mL 0 hour 2_S9_2_Negative (Not Specified) DMSO 0 hour 2_7_200.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_200.0_ug/mL 0 hour 1_7_160.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_160.0_ug/mL 0 hour 1_6_50.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_50.0_ug/mL 0 hour 1_1_Medium DMSO 0 hour 1_8_0.01_ug/mL_Mitomycin-C Mitomycin-C_0.01_ug/mL 0 hour 1_S9_1_Medium DMSO 0 hour 1_S9_5_16.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_16.0_ug/mL 0 hour 2_10_0.01_ug/mL_Mitomycin-C Mitomycin-C_0.01_ug/mL 0 hour 1_S9_8_1.5_ug/mL_Cyclophosphamide Cyclophosphamide_1.5_ug/mL 0 hour 1_S9_4_5.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_5.0_ug/mL 0 hour 2_3_25.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_25.0_ug/mL 0 hour 2_S9_7_200.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_200.0_ug/mL 0 hour 2_2_Medium DMSO 0 hour 2_S9_8_1.5_ug/mL_Cyclophosphamide Cyclophosphamide_1.5_ug/mL 0 hour 2_1_Negative (Not Specified) DMSO 0 hour 1_S9_6_50.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_50.0_ug/mL 0 hour 1_S9_7_160.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_160.0_ug/mL 0 hour 2_S9_6_150.0_ug/mL_C.I. Direct blue 218 C.I. Direct blue 218_150.0_ug/mL ^ASSAY=GENETOX #PARTICIPANT_DISPLAY_NAME #STUDY_TIME #STUDY_TIME_UNIT #PROTOCOL_NAME #SCE_CELLS_EXAMINED #SCE_CHROMOSOMES_EXAMINED #SCE_TOTAL #SCE_PER_CHROMOSOME #SCE_PER_CELL #SCE_PERCENTAGE_INCREASE_OVER_SOLVENT_CONTROL $PARTICIPANT_DISPLAY_NAME $STUDY_TIME $STUDY_TIME_UNIT $PROTOCOL_NAME $SCE_CELLS_EXAMINED $SCE_CHROMOSOMES_EXAMINED $SCE_TOTAL $SCE_PER_CHROMOSOME $SCE_PER_CELL $SCE_PERCENTAGE_INCREASE_OVER_SOLVENT_CONTROL 1_1_Medium 28.0 hour MITOTIC_SHAKE_OFF 50 1039 398 0.383 7.96 0.000 1_2_Negative (Not Specified) 28.0 hour MITOTIC_SHAKE_OFF 50 1039 406 0.391 8.12 2.010 1_3_0.16_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1035 424 0.410 8.48 6.944 1_4_5.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1033 440 0.426 8.80 11.195 1_5_16.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1038 461 0.444 9.22 15.941 1_6_50.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1043 426 0.408 8.52 6.625 1_7_160.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1038 457 0.440 9.14 14.935 1_8_0.01_ug/mL_Mitomycin-C 28.0 hour MITOTIC_SHAKE_OFF 50 1037 2209 2.130 44.18 456.096 2_1_Negative (Not Specified) 28.0 hour MITOTIC_SHAKE_OFF 50 1044 398 0.381 7.96 -4.786 2_2_Medium 28.0 hour MITOTIC_SHAKE_OFF 50 1034 414 0.400 8.28 0.000 2_3_25.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1036 426 0.411 8.52 2.700 2_4_50.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1039 414 0.398 8.28 -0.481 2_5_75.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1038 404 0.389 8.08 -2.792 2_6_100.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1043 488 0.468 9.76 16.857 2_7_200.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1034 561 0.543 11.22 35.507 2_8_300.0_ug/mL_C.I. Direct blue 218 2.0 hour MITOTIC_SHAKE_OFF 0 0 0 0.000 0.00 2_9_0.005_ug/mL_Mitomycin-C 28.0 hour MITOTIC_SHAKE_OFF 50 1047 1430 1.366 28.60 241.122 2_10_0.01_ug/mL_Mitomycin-C 28.0 hour MITOTIC_SHAKE_OFF 50 1046 1854 1.772 37.08 342.689 1_S9_1_Medium 28.0 hour MITOTIC_SHAKE_OFF 50 1036 502 0.485 10.04 0.000 1_S9_2_Negative (Not Specified) 28.0 hour MITOTIC_SHAKE_OFF 50 1042 468 0.449 9.36 -7.310 1_S9_3_1.6_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1042 426 0.409 8.52 -15.628 1_S9_4_5.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1046 434 0.415 8.68 -14.372 1_S9_5_16.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1046 401 0.383 8.02 -20.883 1_S9_6_50.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1042 467 0.448 9.34 -7.508 1_S9_7_160.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1035 509 0.492 10.18 1.492 1_S9_8_1.5_ug/mL_Cyclophosphamide 28.0 hour MITOTIC_SHAKE_OFF 50 1042 1723 1.654 34.46 241.251 2_S9_1_Medium 28.0 hour MITOTIC_SHAKE_OFF 50 1029 406 0.395 8.12 0.000 2_S9_2_Negative (Not Specified) 28.0 hour MITOTIC_SHAKE_OFF 50 1037 402 0.388 8.04 -1.749 2_S9_3_25.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1030 412 0.400 8.24 1.379 2_S9_4_50.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1029 435 0.423 8.70 7.143 2_S9_5_100.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1031 438 0.425 8.76 7.672 2_S9_6_150.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1027 465 0.453 9.30 14.755 2_S9_7_200.0_ug/mL_C.I. Direct blue 218 28.0 hour MITOTIC_SHAKE_OFF 50 1007 454 0.451 9.08 14.266 2_S9_8_1.5_ug/mL_Cyclophosphamide 28.0 hour MITOTIC_SHAKE_OFF 50 1017 1763 1.734 35.26 339.360 ^DATA_TRANSFORMATION=DT_ASSAY_LISTING #DT_ASSAY_NAME #ASSAY_NAME $DT_ASSAY_NAME $ASSAY_NAME SCE SCE_CELLS_EXAMINED SCE SCE_CHROMOSOMES_EXAMINED SCE SCE_TOTAL ^DATA_TRANSFORMATION=DT_SUBJECT_GROUP_LISTING #DT_SUBJECT_GROUP_NAME #GROUP_NAME $DT_SUBJECT_GROUP_NAME $GROUP_NAME 1 1_1_Medium 1 1_2_Negative (Not Specified) 1 1_3_0.16_ug/mL_C.I. Direct blue 218 1 1_4_5.0_ug/mL_C.I. Direct blue 218 1 1_5_16.0_ug/mL_C.I. Direct blue 218 1 1_6_50.0_ug/mL_C.I. Direct blue 218 1 1_7_160.0_ug/mL_C.I. Direct blue 218 1 1_8_0.01_ug/mL_Mitomycin-C 2 2_1_Negative (Not Specified) 2 2_2_Medium 2 2_3_25.0_ug/mL_C.I. Direct blue 218 2 2_4_50.0_ug/mL_C.I. Direct blue 218 2 2_5_75.0_ug/mL_C.I. Direct blue 218 2 2_6_100.0_ug/mL_C.I. Direct blue 218 2 2_7_200.0_ug/mL_C.I. Direct blue 218 2 2_8_300.0_ug/mL_C.I. Direct blue 218 2 2_9_0.005_ug/mL_Mitomycin-C 2 2_10_0.01_ug/mL_Mitomycin-C 1_S9 1_S9_1_Medium 1_S9 1_S9_2_Negative (Not Specified) 1_S9 1_S9_3_1.6_ug/mL_C.I. Direct blue 218 1_S9 1_S9_4_5.0_ug/mL_C.I. Direct blue 218 1_S9 1_S9_5_16.0_ug/mL_C.I. Direct blue 218 1_S9 1_S9_6_50.0_ug/mL_C.I. Direct blue 218 1_S9 1_S9_7_160.0_ug/mL_C.I. Direct blue 218 1_S9 1_S9_8_1.5_ug/mL_Cyclophosphamide 2_S9 2_S9_1_Medium 2_S9 2_S9_2_Negative (Not Specified) 2_S9 2_S9_3_25.0_ug/mL_C.I. Direct blue 218 2_S9 2_S9_4_50.0_ug/mL_C.I. Direct blue 218 2_S9 2_S9_5_100.0_ug/mL_C.I. Direct blue 218 2_S9 2_S9_6_150.0_ug/mL_C.I. Direct blue 218 2_S9 2_S9_7_200.0_ug/mL_C.I. Direct blue 218 2_S9 2_S9_8_1.5_ug/mL_Cyclophosphamide ^DATA_TRANSFORMATION=TREND_ANALYSIS #DT_SUBJECT_GROUP_NAME #DT_ASSAY_NAME #TRIAL_TREND #TRIAL_TREND_P #TRIAL_RESULT $DT_SUBJECT_GROUP_NAME $DT_ASSAY_NAME $TRIAL_TREND $TRIAL_TREND_P $TRIAL_RESULT 1 SCE 1.955 0.025 Negative 2 SCE 4.905 0.000 Weakly Positive 1_S9 SCE 0.715 0.237 Negative 2_S9 SCE 2.591 0.005 Questionable