N-[(1,1,3,3-tetramethylbutyl)phenyl]naphthalen-1-amine

Endpoint summary

Description of key information

In a combined repeated dose toxicity study with the reproduction/developmental toxicity screening test (OECD 422) no adverse findings were noted up to the highest dose tested and the NOAEL was considered to be 1000 mg/kg body weight per day (WIL Research, 2013).

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30 May 2012 - 27 July 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline 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)

Qualifier:

according to guideline

Guideline:

other: The United States Environmental Protection Agency (EPA) Health Effects Test Guidelines, OPPTS 870.3650, Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test, July 2000.

GLP compliance:

yes (incl. QA statement)

Remarks:

WIL Research Europe B.V., Hambakenwetering 7, 5231 DD 's-Hertogenbosch, The Netherlands

Limit test:

no

Species:

rat

Strain:

other: Crl:WI(Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: 11 weeks
- Weight at study initiation: about 200 g (fermales) and 300 g( males)
- Housing:
Pre-mating: Animals were housed in groups of 5 animals/sex/cage in Macrolon plastic cages (MIV type, height 18 cm).
Mating: Females were caged together with males on a one-to-one-basis in Macrolon plastic cages (MIII type, height 18 cm).
Post-mating: Males were housed in their home cage (Macrolon plastic cages, MIV type, height 18 cm) with a maximum of 5 animals/cage. Females wwere individually housed in Macrolon plastic cages (MIII type, height 18 cm).
Lactation: Pups were kept with the dam until termination in Macrolon plastic cages (MIII type, height 18 cm). During locomotor activity monitoring of the dams the pups were kept warm in their home cage using bottles filled with warm water. In order to avoid hypothermia of pups, pups were not left without their dam or a bottle filled with warm water for longer than 30-40 minutes.
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: ad libitum, tap water
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24°C
- Humidity (%): 40 - 70 %
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

propylene glycol

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
Formulations (w/w) were prepared daily within 6 hours prior to dosing and were homogenized to a visually acceptable level. Adjustment was made for specific gravity of the vehicle. No correction was made for the purity of the test substance.

VEHICLE
- Justification for use and choice of vehicle (if other than water): Based on trial formulations performed at WIL Research Europe.
- Amount of vehicle (if gavage): 5 ml/kg

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analyses were conducted on a single occasion during the treatment phase (05 June 2012), according to a validated method (Project 499646, BASF Project 05Y0630/11X472). Samples of formulations were analyzed for homogeneity (highest and lowest concentration) and accuracy of preparation (all concentrations). Stability in vehicle over 6 hours at room temperature was also determined (highest and lowest concentration).
The accuracy of preparation was considered acceptable if the mean measured concentrations were 85-115% of the target concentration. Homogeneity was demonstrated if the coefficient of variation was ≤ 10%. Formulations were considered stable if the relative difference before and after storage was maximally 10%.

Duration of treatment / exposure:

Males were exposed for 29 days, i.e. 2 weeks prior to mating, during mating, and up to the day prior to scheduled necropsy. Females were exposed for 40-54 days, i.e. during 2 weeks prior to mating, during mating, during post-coitum, and during at least 4 days of lactation (up to the day prior to scheduled necropsy). Female nos. 49 and 50 (Group 1), 53 (Group 2), 65 and 66 (Group 3) were not dosed during littering.

Frequency of treatment:

Once daily for 7 days per week, approximately the same time each day with a maximum of 6 hours difference between the earliest and latest dose. Animals were dosed up to the day prior to scheduled necropsy.

Dose / conc.:

62.5 mg/kg bw/day (actual dose received)

Dose / conc.:

250 mg/kg bw/day (actual dose received)

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Dose levels were based on results of a 14-day dose range finding study (Project 499643; BASF Project 01R0630/11X470).

Observations and examinations performed and frequency:

At dose level allocation, 5 animals/sex/group were randomly selected for functional observations, locomotor activity, clinical pathology, organ weights (full list) and histopathology.

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily, detailed clinical observations were conducted for all animals after dosing at no specific time point, but within a similar time period after dosing for the respective animals. Once prior to start of treatment and at weekly intervals during the treatment period this was also performed
outside the home cage in a standard arena.

BODY WEIGHT: Yes
- Time schedule for examinations: Males and females were weighed on the first day of exposure and weekly thereafter. Mated females were weighed on Days 0, 4, 7, 11, 14, 17 and 20 post-coitum and during lactation on Days 1 and 4.

FOOD CONSUMPTION:
Weekly, except for males and females which were housed together for mating and for females without evidence of mating. Food consumption of
mated females was measured on Days 0, 4, 7, 11, 14, 17 and 20 postcoitum and on Days 1 and 4 of lactation.

WATER CONSUMPTION:
Subjective appraisal was maintained during the study, but no quantitative investigation was introduced as no treatment related effect was suspected.

OPHTHALMOSCOPIC EXAMINATION: Yes / No / No data
- Time schedule for examinations:
- Dose groups that were examined:

HAEMATOLOGY: Yes
- Time schedule for collection of blood: on the day of scheduled post mortem examination between 7.00 and 10.30 a.m.
- Anaesthetic used for blood collection: Yes, isoflurane
- Animals fasted: Yes
- How many animals: the selected 5 animals/sex/group
- Parameters checked: White blood cells, Differential leucocyte count, Red blood cells, Reticulocytes, Red blood cell distribution width, Haemoglobin, Haematocrit, Mean corpuscular volume, Mean corpuscular haemoglobin, Mean corpuscular haemoglobin concentration MCHC, Platelets, Prothrombin time, Activated Partial thromboplastin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: on the day of scheduled post mortem examination between 7.00 and 10.30 a.m.
- Animals fasted: Yes
- How many animals: the selected 5 animals/sex/group
- Parameters checked: Alanine aminotransferase, Aspartate aminotransferase, Alkaline phosphatase, Total Protein, Albumin, Total Bilirubin, Urea, Creatinine, Glucose, Cholesterol, Sodium, Potassium, Chloride, Calcium, Inorganic Phosphate, Bile acids

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: The selected males were tested during Week 4 of treatment and the selected females were tested towards the end of the scheduled lactation period after clinical signs observations (incl. arena observations, if applicable) and before blood sampling.
- Dose groups that were examined: the selected 5 animals/sex/group.
- Battery of functions tested: hearing ability, pupillary reflex, static righting reflex, grip strength, locomotor activity

OTHER: General reproduction data: Male number paired with, mating date, confirmation of pregnancy, and delivery day were recorded. Pregnant females were examined to detect signs of difficult or prolonged parturition, and cage debris of pregnant females was examined to detect signs of abortion or premature birth. Any deficiencies in maternal care (such as inadequate construction or cleaning of the nest, pups left scattered and cold, physical abuse of pups or apparently inadequate lactation or feeding) were examined (see section 7.8.1 and 7.8.2 for details).

Sacrifice and pathology:

GROSS PATHOLOGY:
All males and females were deprived of food overnight (with a maximum of 24 hours) prior to planned necropsy, but water was provided. Non-selected females were not deprived of food. Animals surviving to scheduled necropsy were deeply anaesthetized using isoflurane (Abbott B.V., Hoofddorp, The Netherlands) and subsequently exsanguinated. Necropsy was conducted on the following days:
- Females which delivered: on Lactation Days 5 and 7.
- Females which failed to deliver (nos. 64): Post-coitum Days 27 (females with evidence of mating)
- Males: Following completion of the mating period (a minimum of 28 days of dose administration).
All animals were subjected to macroscopic examination of the cranial, thoracic and abdominal tissues and organs, with special attention being paid to the reproductive organs. Descriptions of all macroscopic abnormalities were recorded.

ORGAN WEIGHTS:
The following organ weights and terminal body weight were recorded from the following animals on the scheduled day of necropsy:
- selected 5 animals/sex/group: Adrenal glands, Spleen, Brain, Testes, Epididymides, Thymus, Heart, Uterus (including cervix), Kidneys, Prostate, Liver, Seminal vesicles including coagulating glands, Ovaries, Thyroid including parathyroid
- All remaining males: Epididymides, Testes

HISTOPATHOLOGY:
Samples of the following tissues and organs from all animals were collected and fixed in 10% buffered formalin (neutral phosphate buffered 4% formaldehyde solution, Klinipath, Duiven, The Netherlands):
Ovaries, Adrenal glands, (Pancreas), (Aorta), Peyer's patches [jejunum, ileum] if detectable, Brain - cerebellum, mid-brain, cortex, Pituitary gland, Caecum, Preputial gland, Cervix, Prostate gland, Clitoral gland, Rectum, Colon, (Salivary glands - mandibular, sublingual), Coagulation gland, Sciatic nerve, Duodenum, Seminal vesicles, Epididymides, Skeletal muscle, Eyes (with optic nerve (if detectable) and Harderian gland), (Skin), Spinal cord -cervical, midthoracic, lumbar, (Male and Female mammary gland area), Spleen, Femur including joint, Sternum with bone marrow, Heart, Stomach (forestomach and glandular stomach), Ileum, Testes, Jejunum, Thymus, Kidneys, Thyroid including parathyroid if detectable, (Lacrimal gland, exorbital), (Tongue), (Larynx), Trachea, Liver, Urinary bladder, Lung, infused with formalin, Uterus, Lymph nodes - mandibular, mesenteric, Vagina, (Nasopharynx), All gross lesions, (Esophagus)

Tissues/organs mentioned in parentheses were not examined by the pathologist, since no signs of toxicity were noted at macroscopic examination.

From the selected 5 males of the control and high dose group, and all males suspected to be infertile, additional slides of the testes were prepared to examine staging of spermatogenesis. The testes were processed, sectioned at 3-4 micrometers, and stained with PAS/haematoxylin (Klinipath, Duiven, The Netherlands).

The following slides were examined by a pathologist:
- The preserved organs and tissues of the selected 5 animals/sex of Groups 1 and 4.
- The Additional slides of the testes of all males of Groups 1 and 4 and all males suspected to be infertile or which died before mating to examine staging of spermatogenesis.
- All gross lesions of all animals (all dose groups).
- Kidneys, pituitary and thyroids of all selected males of Groups 2 and 3, based on (possible) treatment-related changes in these organs in Group 4.
- The reproductive organs (cervix, clitoral gland, coagulation gland, epididymides, ovaries, preputial gland, prostate gland, seminal vesicles, testes, uterus, and vagina) of all animals of Groups 1 and 4, and of male 24 (Group 3) that failed to sire and female 64 (Group 3) that failed to deliver healthy pups.

Statistics:

The following statistical methods were used to analyze the data:
- If the variables could be assumed to follow a normal distribution, the Dunnett-test based on a pooled variance estimate was applied for the comparison of the treated groups and the control groups for each sex.
- The Steel-test was applied if the data could not be assumed to follow a normal distribution.
- The Fisher Exact-test was applied to frequency data.
The following additional methods of statistical analysis were used:
Motor activity data was subjected to the Kruskal-Wallis nonparametric ANOVA test to determine intergroup differences followed by the Wilcoxon test to compare the treated groups to the control group.
All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance.
Group means were calculated for continuous data and medians were calculated for discrete data (scores) in the summary tables. Test statistics were calculated on the basis of exact values for means and pooled variances. Individual values, means and standard deviations may have been rounded off before printing. Therefore, two groups may display the same printed means for a given parameter, yet display different test statistics values.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

No clinical signs of toxicity were noted up to 1000 mg/kg bw/day. Salivation was noted for one, one and five animals in the 62.5, 250 and 1000 mg/kg bw/day groups, respectively. This was considered to be a physiological response rather than a sign of systemic toxicity considering the nature and minor severity of the effect and its time of occurrence (i.e. after dosing). This sign may be related to the taste of the test substance, and was not considered to be toxicologically relevant. Incidental findings that were noted included rales (noted for 1 control animal and 1 animal at 1000 mg/kg bw/day), and alopecia. At the incidence observed, these were not considered to be toxicologically relevant.

Mortality:

no mortality observed

Description (incidence):

No mortality occurred during the study period.

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

No toxicologically relevant changes in body weights and body weight gain were noted.
At 1000 mg/kg bw/day, body weight gain was significantly lower for males than controls on Day 8 of the premating period. The difference from controls was only slight and was transient and as such was not considered toxicologically relevant.

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

No toxicologically relevant changes in food consumption before or after allowance for body weight were noted.
Females at 1000 mg/kg bw/day had significantly lower relative food consumption on Days 1-4 of lactation. The difference from controls was only slight and was not considered to reflect treatment related toxicity.

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

No toxicologically relevant changes occurred in haematological parameters of treated rats.
At 1000 mg/kg bw/day, red blood cell distribution width (RDW) was significantly higher than controls. The difference from controls was only slight, and in the absence of effects on any other related haematological parameter like mean corpuscular volume (MCV) and others, this was not considered to be biologically relevant.
At 250 mg/kg bw/day prothrombin time (PT) was significantly lower for males and platelet counts were significantly higher for females, respectively. In the absence of a dose response effect, these were not considered to be treatment related.
Females at 62.5 mg/kg bw/day has significantly increased neutrophil counts with concurrently reduced lymphocyte counts. These differences were attributable to female nos. 57 and 58, and not considered to be treatment-related, since there was no dose dependancy.

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

There were no toxicologically relevant effects on clinical biochemistry parameters noted up to 1000 mg/kg bw/day.
Total bilirubin was increased for animals of both sexes at 250 and 1000 mg/kg bw/day (not statistically significant for males at 250 mg/kg bw/day). In the absence of any relevant effects on the liver noted at the microscopic examination, and since there was no evidence of increased turnover of red blood cells, the increase was not considered biologically relevant.

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

Hearing ability, pupillary reflex, static righting reflex and grip strength were normal in all animals. The variation in motor activity did not indicate a relation with treatment. Both total movements and ambulatory activity were slightly higher for males at 1000 mg/kg bw/day than controls, however, the difference was not significant and in the absence of any relevant clinical signs like hyperactivity, it was not considered to be toxicologically relevant.
All groups showed a similar habituation profile with very high activity in the first interval that decreased over the duration of the test period.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Males at 1000 mg/kg bw/day had slightly, but statistically significantly higher absolute and relative liver weights compared to control animals. In the absence of any corroborative effects seen at the microscopic examination, this was not considered toxicologically relevant. No other organ weight changes were noted.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

There were no toxicologically relevant macroscopic findings noted up to 1000 mg/kg bw/day.
An enlarged spleen was noted for one female at 250 and two females at 1000 mg/kg bw/day, respectively. In the absence of any relevant findings noted at the microscopic examination, this was not considered to be indicative of treatment related toxicity. Incidental findings noted for control and treated animals included reduced size of the seminal vesicles, testes or epididymis, as well as reduced size, reddish discoloration, or many dark red foci on the thymus, pelvic dilation of the right kidney, alopecia of various body areas, uterus contains fluid, diaphragmatic hernia of the left medial lobe of the liver, black focus on the stomach glandular mucosa, and isolated greenish foci on the right clitoral gland. These findings were seen for only a few animals, did not show a treatment related distribution and were thus not considered to be toxicologically relevant.

Neuropathological findings:

not examined

Description (incidence and severity):

Treatment related microscopic findings were noted in the kidneys, pituitary gland and thyroid glands of male rats.
In the kidneys, hyaline droplets in cortical tubules were noted at a minimal or slight degree in five group 1 (0 mg/kg bw/day), five group 2 (62.5 mg/kg bw/day), at a minimal to moderate degree in five group 3 (250 mg/kg bw/day) and five group 4 (1000 mg/kg bw/day) male animals. The hyaline droplets were considered to represent alpha2μglobulin, a normal protein in male rats which undergoes reabsorption in the proximal cortical tubules. A range of chemicals are known to increase hyaline droplet formation. This protein is not present in higher mammals, including man. The degree of moderate recorded in groups 3 (250 mg bw/day) and 4 (1000 mg/kg bw/day) were above spontaneous background levels but were not accompanied by degenerative tubular alterations, and were thus not considered to be adverse.
Multifocal hypertrophy in the adenohypophysis of the pituitary gland was recorded at a minimal degree in three group 1 (0 mg/kg bw/day), in two group 2 (62.5 mg/kg bw/day), three group 3 (250 mg/kg bw/day) and at minimal (2) slight (1) or moderate (1) degree in four group 4 (1000 mg/kg bw/day) male rats. The moderate degree recorded for one male was above the spontaneous severity of minimal or slight for this finding.
In the thyroid glands diffuse follicular hypertrophy/hyperplasia was seen at minimal degree in one group 1 (0 mg/kg bw/day) and at minimal or slight degree in four group 2 (62.5 mg/kg bw/day), four group 3 (250 mg/kg bw/day) and in three group 4 (1000 mg/kg bw/day) animals. This finding at these degrees of severity is a common background finding in male rats of this age i.e. not adverse.
None of the above alterations achieved statistical significance (p < 0.05) in any group when compared to controls, although a trend test revealed a positive trend (p < 0.05) for hyaline droplets in the kidneys.
There were no microscopic findings in any of the animals suspected of infertility which could explain their lack of reproductive performance.
The remaining recorded microscopic findings were within the range of background pathology encountered in Wistar (Han) rats of this age in this type of study and occurred at similar incidences and severity in both control and treated rats. The spermatogenic staging profiles were normal for all animals assessed.

Histopathological findings: neoplastic:

not examined

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: highest dose tested

Critical effects observed:

not specified

Conclusions:

Based on the results presented, a No Observed Adverse Effect Level (NOAEL) of at least 1000 mg/kg bw/day was derived.

Executive summary:

A GLP compliant combined 28-day repeated dose toxicity study with the reproduction/developmental toxicity screening test was performed in male and female Wistar Han rats at dose levels of 62.5, 250 and 1000 mg/kg bw/day. Animals of the control group received the vehicle, propylene glycol, alone. Males were exposed for 29 days, i.e. 2 weeks prior to mating, during mating, and up to termination. Females were exposed for 40-54 days, i.e. during 2 weeks prior to mating, during mating, during postcoitum, and during at least 4 days of lactation. The following observations and examinations were evaluated: mortality / viability, clinical signs (daily), functional observations and locomotor activity (Males: Week 4; females: end of lactation), body weight and food consumption (at least at weekly intervals), clinical pathology (Males: Week 4; females: end of lactation), macroscopy at termination, organ weights and histopathology on a selection of tissues, and reproduction/developmental parameters, consisting of mating, fertility and conception indices, precoital time, number of corpora lutea and implantation sites, gestation index and duration, parturition, maternal care, sex ratio and early postnatal pup development (mortality, clinical signs, body weights and macroscopy). Accuracy, homogeneity and stability of formulations were demonstrated by analyses.

At 1000 mg/kg bw/day, there was an increased severity of multifocal hypertrophy in the adenohypophysis of the pituitary gland in males. The severity (moderate) was higher than background for a single male. No toxicologically relevant changes were noted in any of the remaining parental parameters investigated in this study (i.e. clinical appearance, functional observations, body weight, food consumption, clinical laboratory investigations, macroscopic examination and organ weights).

In conclusion, treatment with the test article by oral gavage in male and female Wistar Han rats at dose levels of 62.5, 250 and 1000 mg/kg body weight/day revealed histological findings at 1000 mg/kg body weight/day characterized by an increased severity of multifocal hypertrophy in the adenohypophysis of the pituitary gland in males only. As this was higher than background in only a single male, when taken together in the absence of any other toxicologically relevant effects, the NOAEL was considered to be 1000 mg/kg bw/day for parental animals.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In a GLP compliant combined repeated dose toxicity study with the reproduction/developmental toxicity screening test (OECD 422), the test article was administered by daily oral gavage to male and female Wistar Han rats at dose levels of 62.5, 250 and 1000 mg/kg bw/day. Animals of the control group received the vehicle, propylene glycol, alone. Males were exposed for 2 weeks prior to mating, during mating, and up to termination (for 29 days). The females were exposed for 2 weeks prior to mating, during mating, during post-coitum, and at least 4 days of lactation (for 40-54 days). Formulation analysis showed that the formulations were prepared accurately, were homogenous, and were stable for at least 6 hours at room temperature.

At 1000 mg/kg body weight per day there was an increased severity of multifocal hypertrophy in the adenohypophysis of the pituitary gland in males. The severity (moderate) was higher than background for a single male. No toxicologically relevant changes were noted in any of the remaining parental parameters investigated in this study (i.e. clinical appearance, functional observations, body weight, food consumption, clinical laboratory investigations, macroscopic examination and organ weights).

In conclusion, treatment with the test article by oral gavage in male and female Wistar Han rats at dose levels of 62.5, 250 and 1000 mg/kg body weight/day revealed histological findings at 1000 mg/kg body weight/day characterized by an increased severity of multifocal hypertrophy in the adenohypophysis of the pituitary gland in males only. As this was higher than background in only a single male, when taken together in the absence of any other toxicologically-relevant effects, the NOAEL was considered to be 1000 mg/kg body weight per day.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
GLP-compliant guideline study

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for the purpose of classification under Regulation (EC) No.1272/2008. Based on the present data, classification for repeated dose toxicity is not warranted under Regulation (EC) No.1272/2008.