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Diss Factsheets

Administrative data

Endpoint:
one-generation reproductive toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
reference to same study
Reference
Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
1986
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
other: Crl:CD (SD)IGS BR
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ORGANISMS
- Age: males 59 days, females 52 days (approximately)
- Number of animals: 10 per sex and per dose group
- Further information: see references
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
ADMINISTRATION / EXPOSURE
- Vehicle: corn oil
- Concentration in vehicle: 6, 20, 60 mg/ml
- Total volume applied: 5 ml/kg bw/dose
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analysis of the test substance indicated that the test substance was homogeneously distributed at all levels and that the concentrations were at the
targed level. Stability results indicated that the test substance was stable at all concentrations.
Duration of treatment / exposure:
males through test day 55; females 4 weeks premating through 4-day lactation period
Frequency of treatment:
daily
Dose / conc.:
30 other: mg/kg bw/day
Dose / conc.:
100 other: mg/kg bw/day
Dose / conc.:
300 other: mg/kg bw/day
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
Post-exposure period: none
MATING PROCEDURES: mating period approximately 1-2 weeks
Observations and examinations performed and frequency:
PARAMETERS ASSESSED DURING STUDY P: 
- Clinical signs: at least once daily
- Mortality: at least twice daily
- Body weight: weekly, for females also on lactation days 0 and 4
- Food consumption: weekly
- Ophthalmoscopic examination: Both eyes of all rats prior to study and  on test day 24, prior to clinical pathology evaluation (surviving rats).
- Clinical pathology evaluation on all rats after 4 weeks of dosing, and  on male rats at the time of scheduled sacrifice:   
- hematology / coagulation: erythrocyte count, total leukocyte count,  platelet count, hemoglobin concentration, hematocrit, differential  
leukocyte count, mean corpuscular volume, mean corpuscular hemoglobin,  mean corpuscular hemoglobin concentration, absolute reticulocyte  counts,  red cell distribution width, microscopic blood examination, activated  partial thromoboplastin time, prothrombin time.   
- clinical chemistry: alkaline phosphatase, alanine aminotransferase,  aspartate aminotransferase, sorbitol dehydrogenase, glucose, urea  nitrogen,  calcium, inorganic phosphorus, total bilirubin, cholesterol,  triglyceride, creatinine, total protein, albumin, globulin, sodium,  potassium, chloride.- urine: volume, specific gravity, urobilinogen, blood, glucose,  protein, appearance (quality, transparency, color), pH, bilirubin,  ketones, sediment  microscopy.
- Other: Neurobehavioral Functional Observational Battery (FOB) on all  study rats prior to exposure and following approximately 4 weeks of test  
substance administration.
OFFSPRING: gestation length, mating index, gestation index, fecundity  index, implantation site numbers, implantation efficiency, sex ratio,  pups 
born alive, viability index
Sacrifice and pathology:
ORGANS EXAMINED AT NECROPSY (MACROSCOPIC AND MICROSCOPIC):
- Time: Sacrifice and necropsy of males on day 56 and of females on days  56-64.
- Organ weights P: liver, kidneys, adrenal glands, thymus, brain, spleen,  heart, testes, epididymides. Calculation of ratios to final body and  brain
 weights.
- Histopathology P:    
all high dose and control rats: testes, epididymides, ovaries, gross  lesions;   
5 high dose and 5 control rats per sex: additional 37 tissues   5 females from other groups with >= 1 offspring: liver
Other examinations:
no other examinations
Statistics:
STATISTICAL METHODS: 
- All weight parameters (P, not F), gestation length, clinical pathology,  grip strength, foot splay: One-way analysis of variance followed with  
Dunnett's test or Kruskal-Wallis test; followed with Dunn's test.
- Incidence of clinical and FOB observations; mating, gestation,  fecundity indexes: Sequential application of Cochran-Armitage test for  trend
- Implantation site number and efficiency, sex ratio, pups born alive,  viability index: Jonckheere's test - Mean pup weights: Linear contrast of the least  square means
- Motor activity: Levene's and Shapiro-Wilk tests followed by repeated  measures analysis of variance followed by contrasts of Jonckheere's trend  test
Clinical signs:
no effects observed
Description (incidence and severity):
There were no test substance-related effects on clinical observations
Mortality:
mortality observed, non-treatment-related
Description (incidence):
One high-dose male was sacrificed in extremis due to a dosing-related injury.
One high-dose female was found dead on day 57 from dystocia.
No other deaths occured during the study.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Decreased body weight gain was observed and considered to be compound-related and of biological significance:
300 mg/kg males (-19 % for days 1-56, not statistically significant; weight -7%);
100 and 300 mg/kg females (only during gestation: gains -13% and -20%, weights -7% and -12%, respectively, statistically significant).
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Increased food consumption was observed and considered to be compound-related and of biological significance:
300 mg/kg males (+19%)
100 mg/kg (+7%, not statistically significant) and 300 mg/kg females (+13%, statistically significant) during gestation.
Food efficiency:
effects observed, treatment-related
Description (incidence and severity):
300 mg/kg males (-33%)
100 and 300 mg/kg females during gestation (+14 and +29%, respectively, statistically significant).
Ophthalmological findings:
no effects observed
Description (incidence and severity):
No test substance-related effects
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
No test substance-related effects in neurobehavioral parameters and motor activity
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Liver weights were statistically significantly increased in several dosed groups:
Males: 30 mg/kg +10% relative; 100 mg/kg +14 % absolute, +21% relative; 300 mg/kg +34 % absolute, +45% relative.
Females: 100 mg/kg +15% relative; 300 mg/kg +36% absolute, +42% relative.
In females it was associated with microscopic centrilobular hepatocellular hypertrophy, which is indicative of a physiological response-induction of enzymes associated with metabolism, and is not considered biologically adverse.

In males, minimal diffuse hypertrophy may have occurred, which is hard to identify histologically.
- Kidney weights were statistically significantly increased in several dosed groups:
Males: 100 mg/kg +16% absolute, +21% relative; 300 mg/kg +22%, +37% relative
Females: 100 mg/kg +11% relative; 300 mg/kg +15% absolute, +17% relative
These findings were associated with some evidence of diuresis in high-dose males and females, but there were no compound-related changes in any other kidney parameter including histopathology. These weight changes may be an adaptive response to the physiological changes that occur following administration of large doses of a test material. They were not considered to be biologically adverse.
Gross pathological findings:
no effects observed
Description (incidence and severity):
No test substance-related effects
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
no effects observed
Description (incidence and severity):
- Fertility index: no test substance-related effects
- Duration of gestation: no test substance-related effects
- Gestation index: no test substance-related effects
- Number of implantations: no test substance-related effects
- Mating index: no test substance-related effects
- Implantation efficiency: no test substance-related effects
- Pups born alive: no test substance-related effects
Details on results:
TOXIC RESPONSE/EFFECTS BY DOSE LEVEL:
- Body weights of pups in the 300 mg/kg group were significantly decreased (-17% on lactation day 4).
- Sex and sex ratios: no test substance-related effects
- Viability index: no test substance-related effects
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
body weight and weight gain
Remarks on result:
other: associated with increased FC and decreased food efficiency
Dose descriptor:
NOAEL
Effect level:
30 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
body weight and weight gain
Critical effects observed:
no
Conclusions:
According to the outcome of the study it is concluded that the no-observed -adverse-effect level (NOAEL) was 100 mg/kg for male rats and 30 mg/kg/day for female rats.
Executive summary:

Male and female rats were administered an oral, daily dose of 0, 30, 100 or 300 mg/kg/day 1,5,9 -cyclododecatriene.

According to the outcome of the study it is concluded that the no-observed -adverse-effect level (NOAEL) was 100 mg/kg for male rats and 30 mg/kg/day for female rats.

Reason / purpose for cross-reference:
reference to same study
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
other: OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
1986
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
other: Crl:CD (SD)IGS BR
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
ADMINISTRATION / EXPOSURE
- Vehicle: corn oil
- Concentration in vehicle: 6, 20, 60 mg/ml
- Total volume applied: 5 ml/kg bw/dose
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analysis of the test substance indicated that the test substance was homogeneously distributed at all levels and that the concentrations were at the
targed level. Stability results indicated that the test substance was stable at all concentrations.
Duration of treatment / exposure:
males through test day 55; females 4 weeks premating through 4-day lactation period
Frequency of treatment:
daily
Dose / conc.:
30 other: mg/kg bw/day
Dose / conc.:
100 other: mg/kg bw/day
Dose / conc.:
300 other: mg/kg bw/day
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
Post-exposure period: none
MATING PROCEDURES: mating period approximately 1-2 weeks
Statistics:
STATISTICAL METHODS: 
- All weight parameters (P, not F), gestation length, clinical pathology,  grip strength, foot splay: One-way analysis of variance followed with  
Dunnett's test or Kruskal-Wallis test; followed with Dunn's test.
- Incidence of clinical and FOB observations; mating, gestation,  fecundity indexes: Sequential application of Cochran-Armitage test for  trend
- Implantation site number and efficiency, sex ratio, pups born alive,  viability index: Jonckheere's test - Mean pup weights: Linear contrast of the least  square means
- Motor activity: Levene's and Shapiro-Wilk tests followed by repeated  measures analysis of variance followed by contrasts of Jonckheere's trend  test
Clinical signs:
no effects observed
Mortality:
mortality observed, non-treatment-related
Description (incidence):
A female in the 300 mg/kg bw/day group was found dead on test day 57 from dystocia. No other deaths occurred.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
During gestation, females administered 100 or 300mg/kg/day had test substance-related, significantly decreased body weight and body weight gain. On gestation day 21, body weights of females in the 100 and. 300mg/kg/day dosage groups were 7 and 12% (statistically significant) below the control mean, respectively .
Body weight gain values for females in the 100 or 300mg/kg/day groups were 13 and 20% below the control mean over the interval of gestation days 0-21, respectively . In addition, weight gain during the interval of gestation days 14-21 was significantly lower compared to the control mean . There were no test substance-related effects on body weight or weight gain for females in the 30 mg/kg/day group .
There were no test substance-related or statistically significant differences in body weight or weight gain during the premating phase for females administered 300 mg/kg/day or below.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
During premating, females in the 300 mg/kg/day group had slightly increased (11%) food consumption compared to the control me an over the interval of test days 1-29 (Table 1). During gestation, a test substance-related, statistically significant increase in food consumption occurred in females administered 300mg/kg/day . Food consumption was significantly increased in females administered 300 mg/kg/d ay during gestation days 7-14, and was increased 13% over the interval of gestation days 0-21 . Food consumption was also increased . 7% in females administered 100 mg/kg/day over the interval of gestation days 0-21.
There were no test substance-related effects or statistically significant differences in food consumption in the 30 mg/kg/day group .
Food efficiency:
effects observed, treatment-related
Description (incidence and severity):
Food efficiency was significantly decreased during gestation days 7-14 for both 100 and 300mg/kg/day females . Over the interval of gestation days 0-21, food efficiency was decreased 14 and 29% in 100 and 300 mg/kg/day females, respectively. The changes in food consumption and food efficiency were associated with changes in body weight during gestation, and were considered to be biologically significant.
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Immunological findings:
not examined
Changes in pregnancy duration:
no effects observed
Other effects:
no effects observed
Description (incidence and severity):
No effects on number of implantation sites
Dose descriptor:
NOAEL
Effect level:
30 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
body weight and weight gain
food consumption and compound intake
food efficiency
Abnormalities:
no effects observed
Fetal body weight changes:
effects observed, treatment-related
Description (incidence and severity):
Body weights of pups in the 300 mg/kg/day group were significantly reduced when compared to control pups on lactation days 0 and 4.
Changes in postnatal survival:
no effects observed
External malformations:
no effects observed
Other effects:
no effects observed
Description (incidence and severity):
No effects on number of pups born
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
fetal/pup body weight changes
Abnormalities:
no effects observed
Developmental effects observed:
no

Data source

Reference
Reference Type:
publication
Title:
Unnamed
Year:
2002

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
1986
GLP compliance:
yes
Limit test:
no

Test material

Constituent 1
Reference substance name:
1,5,9-Cyclododecatriene
IUPAC Name:
1,5,9-Cyclododecatriene
Details on test material:
1,5,9-cyclododecatriene, purity 99.86%

Test animals

Species:
rat
Strain:
other: Crl:CD (SD)IGS BR
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ORGANISMS
- Age: males 59 days, females 52 days (approximately)

Administration / exposure

Route of administration:
oral: gavage
Details on exposure:
ADMINISTRATION / EXPOSURE
- Vehicle: corn oil
- Concentration in vehicle: 6, 20, 60 mg/ml
- Total volume applied: 5 ml/kg bw/dose
Details on mating procedure:
MATING PROCEDURES: 
- mating period approximately 1-2 weeks, beginning after  approximately four weeks of dosing.
Duration of treatment / exposure:
Exposure period: males through test day 55; females 4 weeks premating through 4-day lactation period
Premating exposure period (males): approximately 4 weeks
Premating exposure period (females): approximately 4 weeks
Duration of test: approximately 60 days
Frequency of treatment:
daily
Doses / concentrationsopen allclose all
Dose / conc.:
30 mg/kg bw/day
Dose / conc.:
100 mg/kg bw/day
Dose / conc.:
300 mg/kg bw/day
No. of animals per sex per dose:
Number of animals: 10 per sex and per dose group
Control animals:
yes, concurrent vehicle

Examinations

Parental animals: Observations and examinations:
- Clinical signs: at least once daily
- Mortality: at least twice daily
- Body weight: weekly, for females also on lactation days 0 and 4
- Food consumption: weekly
- Ophthalmoscopic examination: Both eyes of all rats prior to study and  on test day 24, prior to clinical pathology evaluation (surviving rats).
- Clinical pathology evaluation on all rats after 4 weeks of dosing, and  on male rats at the time of scheduled sacrifice:   
- hematology / coagulation: erythrocyte count, total leukocyte count,  platelet count, hemoglobin concentration, hematocrit, differential  leukocyte count, mean corpuscular volume, mean corpuscular hemoglobin,  mean corpuscular hemoglobin concentration, absolute reticulocyte counts,  red cell distribution width, microscopic blood examination, activated  partial thromoboplastin time, prothrombin time.   
- clinical chemistry: alkaline phosphatase, alanine aminotransferase,  aspartate aminotransferase, sorbitol dehydrogenase, glucose, urea  nitrogen, calcium, inorganic phosphorus, total bilirubin, cholesterol,  triglyceride, creatinine, total protein, albumin, globulin, sodium,  potassium, chloride.  
- urine: volume, specific gravity, urobilinogen, blood, glucose,  protein, appearance (quality, transparency, color), pH, bilirubin,  ketones, sediment microscopy.
- Other: Neurobehavioral Functional Observational Battery (FOB) on all  study rats prior to exposure and following approximately 4 weeks of test  substance administration.
Litter observations:
OFFSPRING: gestation length, mating index, gestation index, fecundity  index, implantation site numbers, implantation efficiency, sex ratio,  pups born alive, viability index
Postmortem examinations (parental animals):
ORGANS EXAMINED AT NECROPSY (MACROSCOPIC AND MICROSCOPIC):
- Time: Sacrifice and necropsy of males on day 56 and of females on days  56-64.
- Organ weights P: liver, kidneys, adrenal glands, thymus, brain, spleen,  heart, testes, epididymides. Calculation of ratios to final body and  brain weights.
- Histopathology P:    all high dose and control rats: testes, epididymides, ovaries, gross  lesions;   5 high dose and 5 control rats per sex: additional 37 tissues   5 females from other groups with >= 1 offspring: liver
Postmortem examinations (offspring):
Implantation site number and efficiency, sex ratio, mean pup weights, pups born alive,  viability index
Statistics:
STATISTICAL METHODS: 
- All weight parameters (P, not F), gestation length, clinical pathology,  grip strength, foot splay: One-way analysis of variance followed with  Dunnett's test or Kruskal-Wallis test; followed with Dunn's test.
- Incidence of clinical and FOB observations; mating, gestation,  fecundity indexes: Sequential application of Cochran-Armitage test for  trend
- Implantation site number and efficiency, sex ratio, pups born alive,  viability index: Jonckheere's test
- Mean pup weights: Linear contrast of the least square means
- Motor activity: Levene's and Shapiro-Wilk tests followed by repeated  measures analysis of variance followed by contrasts of Jonckheere's trend  test.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
Description (incidence and severity):
There were no test substance-related effects on clinical observations
Mortality:
mortality observed, non-treatment-related
Description (incidence):
One high-dose male was sacrificed in extremis due to a dosing-related injury. One high-dose female was found dead on day 57 from dystocia. No other deaths occured during the study.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Decreased body weight gain was observed and considered to be compound-related and of biological significance: 300 mg/kg males (-19 % for days 1-56, not statistically significant; weight -7%); 100 and 300 mg/kg females (only during gestation: gains -13% and -20%, weights -7% and -12%, respectively, statistically significant).
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Increased food consumption was observed and considered to be compound-related and of biological significance: 300 mg/kg males (+19%) 100 mg/kg (+7%, not statistically significant) and 300 mg/kg females (+13%, statistically significant) during gestation.
Food efficiency:
effects observed, treatment-related
Description (incidence and severity):
As a consequence, food efficiency was significantly reduced: 300 mg/kg males (-33%) 100 and 300 mg/kg females during gestation (+14 and +29%, respectively, statistically significant).
Ophthalmological findings:
no effects observed
Description (incidence and severity):
no test substance-related effects
Haematological findings:
no effects observed
Description (incidence and severity):
no test substance-related effects
Clinical biochemistry findings:
no effects observed
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
There were no test substance-related effects in neurobehavioral parameters or motor activity.
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
no test substance-related adverse effects

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
Description (incidence and severity):
- Fertility index: no test substance-related effects
- Duration of gestation: no test substance-related effects
- Gestation index: no test substance-related effects
- Number of implantations: no test substance-related effects
- Mating index: no test substance-related effects;
- Implantation efficiency: no test substance-related effects

Details on results (P0)

NOAEL (NOEL), LOAEL (LOEL):    
- NOAEL  30 mg/kg for females, 100 mg/kg for males    
- LOAEL 100 mg/kg for females, 300 mg/kg for males 

Effect levels (P0)

open allclose all
Dose descriptor:
NOAEL
Effect level:
300 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
reproductive performance
other: no adverse effects observed
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day
Sex:
male
Basis for effect level:
body weight and weight gain
food consumption and compound intake
food efficiency
Dose descriptor:
NOAEL
Effect level:
30 mg/kg bw/day
Sex:
female
Basis for effect level:
body weight and weight gain
food consumption and compound intake
food efficiency

Target system / organ toxicity (P0)

Critical effects observed:
no

Results: F1 generation

General toxicity (F1)

Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Body weights of pups in the 300 mg/kg group were significantly decreased (-17% on lactation day 4)
Other effects:
no effects observed
Description (incidence and severity):
- Sex and sex ratios: no test substance-related effects
- Pups born alive: no test substance-related effects

Details on results (F1)

- NOAEL  100 mg/kg for pups  
- LOAEL  300 mg/kg for pups

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
100 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain

Target system / organ toxicity (F1)

Critical effects observed:
no

Overall reproductive toxicity

Reproductive effects observed:
no

Any other information on results incl. tables

Summary of reproductive outcomes:

 

- Dose (mg/kg) 0 30 100 300

- Mating Index(%): 80.0 90.0 100.0 100.0

- Fertility Index (%): 87.5 77.8 70.0 70.0

- Gestation Length (days): 22.0 22.0 22.0 22.0

- Implantations (mean/litter): 16.0 15.6 16.3 16.1

- Implantation efficiency (%): 92.1 96.2 92.0 89.5

- Gestation Index: 100.0 100.0 100.0 100.0

- Mean % Born Alive: 98.3 99.1 99.2 99.0

- 0-4 Day Viability (%): 98.0 99.0 98.2 98.3

- Sex Ratio (males): 0.45 0.50 0.49 0.45

Applicant's summary and conclusion