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EC number: 200-607-2 | CAS number: 65-31-6
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Endpoint summary
Administrative data
Key value for chemical safety assessment
- Toxic effect type:
- dose-dependent
Effects on fertility
Description of key information
Reproductive toxicity
Based on the experimental studies of test chemical Low Observed Adverse Effect Level (LOAEL) was considered to be 1.0mg/kg bw. Thus, comparing this value with the criteria of CLP regulation test chemical can be classified as “Category 2”for reproductive toxicity
Link to relevant study records
- Endpoint:
- screening for reproductive / developmental toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- secondary literature
- Justification for type of information:
- Data from peer reviewed journal
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- other: As mentioned below
- Principles of method if other than guideline:
- Reproductive toxicity study of test material was performed on male albino rats.
- GLP compliance:
- not specified
- Limit test:
- no
- Justification for study design:
- No data available
- Species:
- rat
- Strain:
- other: albino rats
- Details on species / strain selection:
- No data available
- Sex:
- male
- Details on test animals or test system and environmental conditions:
- Details on test animals and env. conditions
TEST ANIMALS
- Source: Animal House, College of Medicine,
University of Ibadan, Oyo State, Nigeria
- Age at study initiation: 8-10 weeks old
- Weight at study initiation: average weight ranged between 150 and 180 g
- Fasting period before study:
- Housing:
- Use of restrainers for preventing ingestion (if dermal):
yes/no
- Diet (e.g. ad libitum): ad libitum access to rat chow food
- Water (e.g. ad libitum): ad libitum access to drinking water.
- Acclimation period:
ENVIRONMENTAL CONDITIONS
- Photoperiod (hrs dark / hrs light): 12:12‑hour light–dark - Route of administration:
- oral: gavage
- Vehicle:
- physiological saline
- Details on exposure:
- Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Test material dissolved in normal saline
DIET PREPARATION
- Rate of preparation of diet (frequency):No data available
- Mixing appropriate amounts with (Type of food )
- Storage temperature of food: No data available
VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle: 0, 1.0 mg/kg bw/day
- Amount of vehicle (if gavage): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available
Other: The working solutions were stored in foil‑wrapped glass bottle at 4°C for no longer than 10 days - Details on mating procedure:
- - M/F ratio per cage:4:5
- Length of cohabitation: 5days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy:The presence of a vaginal plug was accepted as the index for a positive mate and taken as day one of pregnancy
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: [no / yes (explain)]
- After successful mating each pregnant female was caged (how):
- Any other deviations from standard protocol: - Analytical verification of doses or concentrations:
- not specified
- Duration of treatment / exposure:
- 30 days
- Frequency of treatment:
- Daily
- Details on study schedule:
- No data available
- Remarks:
- 0, 1.0mg/kg bw/day
- No. of animals per sex per dose:
- 32 male rats
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- No data available
- Positive control:
- No data available
- Parental animals: Observations and examinations:
- Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes
DETAILED CLINICAL OBSERVATIONS: Yes
Time schedule: daily
BODY WEIGHT: Yes
Time schedule for examinations: daily
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes Food consumption was determined weekly.
Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day:
Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data: No data available
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
Time schedule for examinations:
OTHER: - Oestrous cyclicity (parental animals):
- No data available
- Sperm parameters (parental animals):
- Parameters examined in {P} male parental generations:
testis weight, epididymis weight, daily sperm production, sperm count in testes, sperm count in epididymides, , sperm motility, sperm morphology, were observed - Litter observations:
- The number of litters delivered and their body weights were determined.
- Postmortem examinations (parental animals):
- No data available
- Postmortem examinations (offspring):
- No data available
- Statistics:
- Data obtained were expressed in Mean ± SEM. Statistical analysis was performed by analysis of variance (ANOVA) followed by multiple comparison by two‑tailed t‑test. The values for P < 0.05 were considered to be statistically significant.
- Reproductive indices:
- No data available
- Offspring viability indices:
- No data available
- Clinical signs:
- not specified
- Dermal irritation (if dermal study):
- not specified
- Mortality:
- not specified
- Body weight and weight changes:
- not specified
- Food consumption and compound intake (if feeding study):
- not specified
- Food efficiency:
- not specified
- Water consumption and compound intake (if drinking water study):
- not specified
- Ophthalmological findings:
- not specified
- Haematological findings:
- not specified
- Clinical biochemistry findings:
- not specified
- Urinalysis findings:
- not specified
- Behaviour (functional findings):
- not specified
- Immunological findings:
- not specified
- Organ weight findings including organ / body weight ratios:
- not specified
- Histopathological findings: non-neoplastic:
- not specified
- Histopathological findings: neoplastic:
- not specified
- Other effects:
- not specified
- Reproductive function: oestrous cycle:
- not specified
- Reproductive function: sperm measures:
- effects observed, treatment-related
- Description (incidence and severity):
- Oral administration of 1.0 mg/kg BW of test material on animals daily for a period of 4 weeks significantly
decrease (P < 0.05) the progressive motility of the sperm when compared with the control group
The mean epididymal sperm count of animals administered with 1.0 mg/kg BW was significantly decreased (P < 0.05) when compared with their control
A significant decrease (P > 0.05) was recorded for the mean percentage live sperm of rats treated with 1.0 mg/k bw when compared with the control
The test material caused an insignificant decrease (P > 0.05) in the epididymal sperm volume in the treated groups when compared with the control
Administration of 1.0 mg/kg bw test material significantly reduced normal sperm morphology
The mean serum testosterone level of animals that received 1.0 mg/kg bw of nicotine was significantly decreased (P < 0.05) when compared with the control group. - Reproductive performance:
- effects observed, treatment-related
- Description (incidence and severity):
- Administration of 1.0 mg/kg bw test material caused significant decrease in libido score when compared with the control, test material significantly decreased percentage fertility when compared with the control
- Dose descriptor:
- LOAEL
- Effect level:
- 1 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male
- Basis for effect level:
- reproductive function (sperm measures)
- reproductive performance
- Remarks on result:
- other: effects observed treatment related
- Critical effects observed:
- not specified
- System:
- other: not specified
- Organ:
- not specified
- Clinical signs:
- not specified
- Dermal irritation (if dermal study):
- not specified
- Mortality / viability:
- not specified
- Body weight and weight changes:
- not specified
- Food consumption and compound intake (if feeding study):
- not specified
- Food efficiency:
- not specified
- Water consumption and compound intake (if drinking water study):
- not specified
- Ophthalmological findings:
- not specified
- Haematological findings:
- not specified
- Clinical biochemistry findings:
- not specified
- Urinalysis findings:
- not specified
- Sexual maturation:
- not specified
- Organ weight findings including organ / body weight ratios:
- not specified
- Gross pathological findings:
- not specified
- Histopathological findings:
- not specified
- Other effects:
- not specified
- Behaviour (functional findings):
- not specified
- Developmental immunotoxicity:
- not specified
- Dose descriptor:
- other: not specified
- Generation:
- other: not specified
- Based on:
- test mat.
- Sex:
- not specified
- Basis for effect level:
- other: not specified
- Remarks on result:
- not measured/tested
- Critical effects observed:
- not specified
- System:
- other: not specified
- Organ:
- not specified
- Reproductive effects observed:
- not specified
- Treatment related:
- not specified
- Conclusions:
- Low Observed Adverse Effect Level (LOAEL) was considered to be 1.0 mg/kg/day, When male albino rats were treated with test material orally.
- Executive summary:
Sperm function and fertility profile of test material was performed on 32 male albino rats whose average weight ranged between 150 and 180 g (8-10 weeks old). The test material dosage freshly prepared in normal saline for each group of animals was delivered orally at 1.0 mg/kg body weight (BW). The working solutions were stored in foil‑wrapped glass bottle at 4°C for no longer than10 days. A total of 20 untreated fertile, Proestrum female rats were used for the fertility test. Five untreated female rats were cohabited with each other of the four male groups from the day 31 of treatment. All animals were cohabited for 5 days. The presence of a vaginal plug was accepted as the index for a positive mate and taken as day one of pregnancy. The number of litters delivered and their body weights were determined. Oral administration of 1.0 mg/kg BW of test material on animals daily for a period of 4 weeks significantly decrease (P< 0.05) the progressive motility of the sperm when compared with the control group. The mean epididymal sperm count of animals administered with 1.0 mg/kg BW was significantly decreased (P< 0.05) when compared with their control. A significant decrease (P> 0.05) was recorded for the mean percentage live sperm of rats treated with 1.0 mg/k bw when compared with the control. The test material caused an insignificant decrease (P> 0.05) in the epididymal sperm volume in the treated groups when compared with the control. A significant decrease (P> 0.05) was recorded for the mean percentage live sperm of rats treated with 1.0 mg/k bw when compared with the control. The test material caused an insignificant decrease (P> 0.05) in the epididymal sperm volume in the treated groups when compared with the control. Administration of 1.0 mg/kg bw test material significantly reduced normal sperm morphology. The mean serum testosterone level of animals that received 1.0 mg/kg bw of nicotine was significantly decreased (P< 0.05) when compared with the control group.
Reference
Semen parameters of experimental rats treated with test material
Dose |
Motility (%) |
Live/dead (%) |
Volume (ml) |
Count (106/ml) |
ratio |
|
|
|
|
Control |
85.50±3.21 |
94.56±4.21 |
5.22±0.04 |
112.40±10.40 |
Nicotine |
|
|
|
|
1.0 mg/kg+ |
32.00±4.65* |
79.80±5.17* |
5.18±0.06 |
64.20±5.60* |
Values are expressed as means ± S.E.M of 8 rats per group. *P<0.05 vs control
Fertility profile of experimental rats treated with test material
Dose |
Libido score |
Litter number |
Litter weight (g) |
Percentage fertility |
Control |
8.82 ± 1.21 |
6.24 ±040 |
5.96± 0.20 |
100 |
1.0 mg/kg+ test material |
3.26 ±1.62* |
0.00 ±0.00* |
0.00 ±0.00* |
0* |
Values are expressed as means ± S.E.M of 8 rats per group. *P< 0.05 vs control
Effect on fertility: via oral route
- Endpoint conclusion:
- adverse effect observed
- Dose descriptor:
- LOAEL
- 1 mg/kg bw/day
- Study duration:
- subacute
- Species:
- rat
- Quality of whole database:
- Data from peer reviewed journal
Effect on fertility: via inhalation route
- Endpoint conclusion:
- no study available
Effect on fertility: via dermal route
- Endpoint conclusion:
- no study available
Additional information
Reproductive toxicity
In different studies of test chemical has been investigated for reproductive toxicity to a greater or lesser extent. Often are the studies based on in vivo experiments i.e. most commonly in rats for test chemical.The studies are as mentioned below:
Study 1
Sperm function and fertility profile of test material was performed on32 male albino rats whose average weight ranged between 150 and 180 g (8-10 weeks old). The test material dosage freshly prepared in normal saline for each group of animals was delivered orally at 1.0 mg/kg body weight (BW). The working solutions were stored in foil‑wrapped glass bottle at 4°C for no longer than10 days. A total of 20 untreated fertile, Proestrum female rats were used for the fertility test. Five untreated female rats were cohabited with each other of the four male groups from the day 31 of treatment. All animals were cohabited for 5 days. The presence of a vaginal plug was accepted as the index for a positive mate and taken as day one of pregnancy. The number of litters delivered and their body weights were determined. Oral administration of 1.0 mg/kg BW of test material on animals daily for a period of 4 weeks significantly decrease (P< 0.05) the progressive motility of the sperm when compared with the control group. The mean epididymal sperm count of animals administered with 1.0 mg/kg BW was significantly decreased (P< 0.05) when compared with their control. A significant decrease (P> 0.05) was recorded for the mean percentage live sperm of rats treated with 1.0 mg/k bw when compared with the control. The test material caused an insignificant decrease (P> 0.05) in the epididymal sperm volume in the treated groups when compared with the control. A significant decrease (P> 0.05) was recorded for the mean percentage live sperm of rats treated with 1.0 mg/k bw when compared with the control. The test material caused an insignificant decrease (P> 0.05) in the epididymal sperm volume in the treated groups when compared with the control. Administration of 1.0 mg/kg bw test material significantly reduced normal sperm morphology. The mean serum testosterone level of animals that received 1.0 mg/kg bw of nicotine was significantly decreased (P< 0.05) when compared with the control group.
Study 2
The reproductive toxicity study of test material was performed on56 albino wistar rats (42 female, 14 male). The test material was dissolved in saline in dose concentration 0.5,1.0mg/g bw orally for forty-three days. After 30 days of treatment, male rats were introduced to the female in ratio 1:3, (i.e. 1 male to 3 female). The vaginal smears were examined daily in the morning between 8-10a.m for the presence of sperm plug, which indicates positive copulation. The animals which showed thick clumps of spermatozoa in the vaginal smear were separated and that day was designated as Day 1 of pregnancy. Body weight of the animals was monitored weekly throughout the period of the study. On the 13th day of pregnancy, the animals were injected 0.3ml of 0.5% Evans blue dye through the tail vein, stabilized for 15 minutes post injection and Blood samples were collected through cardiac puncture into heparinized tubes at 4000rmfor 15 minutes. Plasma was collected into plain tubes for estrogen and progesterone assay using Enzyme Linked Immunosorbent Assay (ELISA) method The animals were dissected and uteri were opened to ascertain implantation sites The dye sites were counted and recorded as implantation sites per rat. The brain, heart, kidneys, liver, lungs, uterine tubes, ovaries and vagina were harvested and weighed.The growth rate of the animals in groups 2 to 7 was significantly reduced when compared with control (p<0.05). There was significant increase in plasma estrogen level in 0.5 and 1.0 mg/kg bw dose group when compared with the control (p<0.05). Also, the plasma progesterone was significantly increased in 1.0 mg/kg dose group relative to the control (p<0.05). The ratio of estrogen to progesterone was significantly increased (p<0.05) in 0.5 and 1.0 mg/kg bw dose group when compared with the control. The test material have no significant
effect on ovaries and uterine tube weight (p>0.05), while the vagina weight was significantly reduced in rats treated with 1.0mg/kg bw dose of test material and it recovery group when compared with control (p<0.05).There was a significant decrease in implantation site of pregnancy rats that were treated with 0.5 and 1.0 mg/kg bw dose of test material when compared with the control (p<0.05). Similarly, the implantation site in recovery groups was significantly decreased when compared with the control (p<0.05). There was no significant difference in the brain, kidneys, and liver weight of rats in all the groups (p>0.05), while heart and lungs weight showed significant reduction in 0.5 and 1.0 mg/kg bw dose group when compared with control (p<0.05).Hence Low Observed Adverse Effect Level (LOAEL) was considered to be 1.0 mg/kg/day, When female and male wistar rats were treated with test material orally.
Study 3
Sperm function and fertility profile of test material was performed on forty male and twenty-five female Sprague-Dawley rats, 2 - 2.5 month old and whose average weight ranged between 150gand 180g,The male animals in the five groups were treated for 30 days and they included the control group that received 0.2ml/kg normal saline, 0.5mg/kg nicotine-treated group, 1.0mg/kg nicotine-treated group, 0.5mg/kg nicotine-treated group but left untreated for another 30 days and 1.0mg/kg nicotine-treated group but left untreated for another 30 days. A total of 25 untreated fertile, prestrous female rats were used for the fertility test. Five untreated female rats were cohabited with a male rat from one of the five male groups on the 31 day of treatment except the recovery groups whose cohabitation commenced on the 31 day of the recovery period. All animals were cohabited for 5 days. The presence of a vaginal plug was accepted as the index for a positive mating and it was taken as day one of pregnancy. The number of litters delivered and their body weights were determined. Daily oral test material administration of 0.5mg/kg and 1.0mg/kg per body weight for a period of four weeks significantly decreased (P < 0.05) the progressive motility of the sperm when compared with the control group. The mean sperm counts per epididymal volume of rats administered with 0.5mg/kgand 1.0mg/kg body weight significantly decreased (P < 0.05) when compared with the controls. This decrease was dose-dependent. There was also a decrease in the mean epididymal sperm counts of the recovery groups for 0.5mg/kg and 1.0mg/kg B.W. An insignificant decrease (P > 0.05) was recorded for the mean percentage of live sperms in rats treated with both 0.5mg/kgand 1.0mg/kg body weight treated groups when compared with the controls.
The most common abnormality encountered during the morphological examination of the sperms in the rats that received the two daily doses of nicotine was the “curve tail” which accounted for 60% of the observed abnormalities. Though, there seemed to be a dose-dependent morphological abnormality. The observed “curve tail” abnormality was statistically significant (P < 0.05) when compared with the controls. a significant decrease (P < 0.01) in their libido when compared with their control counterparts. The observed decrease was dose-dependent. The results showed that test material caused an insignificant decrease (P > 0.05) in the epididymal volume for both 0.5mg/kg and 10.0mg/kg B.W. treated groups when compared with the control group. The female rats used for mating in the control group had 100% fertility rate while 0.5mg/kg and 1.0mg/kg B.W. treated rats had 40% and 0% fertility rates, respectively. The recovery group for 0.5mg/kg B.W. had a fertility rate of 80% while the recovery group for 1.0mg/kgB.W. had a fertility rate of 60% .Female rats cohabited with male rats from the 1.0 mg/kg bw dose group did not conceive throughout the study period. However, these effects were reversible after withdrawn of test material from the rats. Female rats used for mating in the control group had an average litter size of 7.80 ± 0.4. An average litter size of 4.50 ± 0.5 was produced by female rats used in mating the 0.5mg/kg B.W. treated animals. The recovery groups of 0.5mg/kg B.W. and 1.0mg/kg B.W. treated rats had an average litter size of 6.33 ± 0.5 and 3.50 ± 0.5, respectively. Based on study test material reduces fertility in adult male rats and adversely affect litters’ weight and size delivered by untreated female rats. The data also indicate that test material withdrawal for a particular period of time could ameliorate the observed effects. Hence Low Observed Adverse Effect Level (LOAEL) was considered to be 1.0 mg/kg/day, When male Sprague Dawley rats were treated with test material orally.
Study 4
Effects of Oral Administration of test material on Organ Weight, Serum Testosterone Level and Testicular Histology was performed in adult male rats.40 male Sprague-Dawley rats whose average weight ranged between 150g and 180g (2-2.5 month old) were used in study. The animals were divided into five groups; Control group that received 0.2 ml/kg normal saline, 0.5mg/kg and 1.0mg/kg treated group while 0.5mg/kg and 1.0mg/kg test material treated but left untreated for another 30 days. Control group received 0.2 ml of normal saline (vehicle) which was used to dissolve test material and administered daily by oral gavage through the use of oral cannula. The animals were dissected and the reproductive organs (testes, epididymis, prostate gland and seminal vesicle) were removed, cleared of adherent tissues and weighed immediately with an electronic weighing balance, model DT 1000 England with a capacity of 0.1 to 1000g. Testes and epididymis of the control and treated rats were fixed in Bouinʼs fluid for 6 hours before they were transferred into 10% formalin for histological evaluation. The tissues were routinely processed and examined under the light microscope. Photomicrograph of the slide was then taken.
There were no significant differences in the mean body weight of treated rats during the treatment period compared with the control group, the mean serum testosterone level in rats that received 0.5 mg/kg B.W (low dose) and those that received 1.0 mg/kg B.W (high dose) of test material for four weeks was significantly decreased (p<0.05) in dose-dependent manner when compared with the control group. This decrease is dose dependent. The result showed that there is a significant decrease in the mean testicular weight of rats that received 0.5mg/kg B.W and 1.0mg/kg B.W in a dose-dependent manner.The mean epididymal weight of rats that received the two doses of test material was significantly decreased (p<0.05) when compared with the control. Their recovery groups showed no significant decrease in the mean epididymal weight when compared with their control. Rats treated with 0.5mg/kg and 1.0mg/kg BW of test material for four weeks showed no significant increase in the mean prostate weight when compared with the control while an insignificant decrease in the mean prostate weight of 0.5mg/kg recovery group. Rats treated with 0.5mg/kg and 1.0mg/kg B.W of test material had an insignificant decrease in the mean seminal vesicle weight, There was a reduction in the germ cells and sertoli cell population of nicotine treated groups. The severity of testicular degeneration and disruption of the interstitum was dose-dependent, as observed changes were more pronounced in the high dose group than the low dose group. However, there were both regeneration of the germinal epithelium and restructuring of the interstitum towards normal in the recovery group. The histological study shows that nicotine had no effect on the epididymis of the animals. Hence Low Observed Adverse Effect Level (NOAEL) was considered to be 1.0 mg/kg/day, as decrease serum testosterone level and have deleterious effect on the male reproductive organ and histology of albino rats were treated with test material orally.
Thus, based on the above experimental studies of test chemical Low Observed Adverse Effect Level (LOAEL) was considered to be 1.0mg/kg bw Thus, comparing this value with the criteria of CLP regulation test chemical can be classified as reproductive toxicant.
Effects on developmental toxicity
Effect on developmental toxicity: via oral route
- Endpoint conclusion:
- no study available
Effect on developmental toxicity: via inhalation route
- Endpoint conclusion:
- no study available
Effect on developmental toxicity: via dermal route
- Endpoint conclusion:
- no study available
Justification for classification or non-classification
Thus, comparing this value with the criteria of CLP regulation test chemical can be classified as “Category 2”for reproductive toxicity
Additional information
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