Registration Dossier

Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
07 February 2017 to 12 June 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Version / remarks:
1998
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 7-8 weeks old
- Weight at study initiation: males: 152 – 187 g; females: 122 – 151 g
- Fasting period before study: none
- Housing: - The animals were kept in groups of 5 animals / sex / group / cage in IVC cages on Altromin saw fibre bedding
- Diet (e.g. ad libitum): Altromin 1324 maintenance diet for rats and mice, ad libitum
- Water (e.g. ad libitum): tap water, sulphur acidified to a pH of approximately 2.8, ad libitum
- Acclimation period: at least 5 days

DETAILS OF FOOD AND WATER QUALITY: not specified

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3 °C
- Humidity (%): 55 +/- 10 %
- Air changes (per hr): 10 x / hour
- Photoperiod (hrs dark / hrs light): 12 hours light, 12 hours dark

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: The test item and control formulations were prepared once in 7-10 days based on the stability information. The prepared formulation was stored at room temperature. The test item was weighed on a suitable precision balance into a tared plastic vial and the vehicle was added to give the appropriate final concentration of the test item. To protect from hydrolysis by humidity, formulation vials were purged with argon after the preparation before closing. Formulates were kept under magnetic stirring during the daily administration.

VEHICLE
- Justification for use and choice of vehicle (if other than water): The vehicle has been selected as suggested by the sponsor based on the test item’s characteristics (hydrolysis in water) and testing guideline. The selected vehicle was dried corn oil.
- Concentration in vehicle: 2.53, 12.63, 25.25 mg/mL
- Amount of vehicle (if gavage): not specified
- Lot/batch no. (if required): MKBQ9948V
- Purity: not specified
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Before beginning of the treatment period, formulation samples were prepared and analysed in order to obtain knowledge about stability and homogeneity of the test item in the selected vehicle. Study pre-start stability analysis included samples from high dose and low dose group and the investigation was made. Prestart homogeneity investigation included the samples collected from various levels (top, middle and bottom) of high dose and low dose groups (6 samples).
Duration of treatment / exposure:
90 days
Frequency of treatment:
7 days a week
Doses / concentrationsopen allclose all
Dose / conc.:
10 mg/kg bw/day (actual dose received)
Remarks:
low dose group (LD)
Dose / conc.:
50 mg/kg bw/day (actual dose received)
Remarks:
middle dose group (MD)
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Remarks:
high dose group (HD)
No. of animals per sex per dose:
10 male and 10 female animals per group
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose levels were selected according to the results of a previous toxicity studies and in consultation with the sponsor.
- Rationale for animal assignment (if not random): random
- Rationale for selecting satellite groups: Recovery groups of 5 animals from control and high dose group.
- Post-exposure recovery period in satellite groups: 28 days
- Section schedule rationale (if not random): random
Positive control:
not used

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
- Cage side observations checked included: clinical signs of toxicity, mortality and morbidity

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Once before the first administration and at least once a week thereafter

BODY WEIGHT: Yes
- Time schedule for examinations: Once before the assignment to the experimental groups, on the first day of administration and weekly during the treatment and recovery period.

FOOD CONSUMPTION AND COMPOUND INTAKE: Food consumption was measured weekly.

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Not applicable

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: prior to sacrifice
- Anaesthetic used for blood collection: No
- Animals fasted: Yes
- How many animals: all the animals
- Parameters checked in table [No.1] were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: prior to sacrifice
- Animals fasted: Yes
- How many animals: all the animals
- Parameters checked in table [No.1] were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: prior to sacrifice
- Metabolism cages used for collection of urine: No
- Animals fasted: Yes
- Parameters checked in table [No.1] were examined.

NEUROBEHAVIOURAL EXAMINATION: Yes
- Dose groups that were examined: all animals
- Battery of functions tested: sensory activity / grip strength / motor activity

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes (see table No 2)

HISTOPATHOLOGY: Yes (see table No 2)
Other examinations:
Immunohistochemistry of α2µ-globulin and/or a specific staining was performed on kidney of all male animals sacrificed at the end of the treatment period of the study (40 animals) and recovery period (10 animals).
Statistics:
A statistical assessment of the results of the body weight, food consumption, parameters of haematology, blood coagulation and clinical biochemistry and absolute and relative organ weights was performed for each gender by comparing values of dosed with control animals using either a parametric one-way ANOVA and a post-hoc Dunnett Test or a non-parametric Kruskal-Wallis Test and a post-hoc Dunn’s Test, based on the results of homogeneity and normality tests. Furthermore, statistical comparisons of data acquired during the recovery period were performed with a Student’s t-Test or Mann-Whitney U-Test when appropriate. These statistics were performed with Ascentos 1.1.3 software or GraphPad Prism V.6.01 software (p<0.05 is considered as statistically significant).

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
In this study, there were no clinical findings observed related to a systemic effect of the test item.
During the treatment period, slight to moderate salivation was noted occasionally in one male of LD group, some males and females of the MD group and the HD group. Moving the bedding was observed transitorily in all males and all females of the MD and HD group. As the symptoms of salivation and moving the bedding were noted mainly immediately after administration, these signs were considered to be a sign of discomfort due to a local reaction to the test item rather than the systemic effect of the test item. No clinical signs were observed in any recovery animals during recovery period except alopecia on lower back in one HD recovery female from day 111 to 118 was observed.
Low incidences of slight clinical signs like alopecia on various body parts, skin bruising, skin scratches/cuts on various body parts were noted in isolated males and/or females of the dose groups during the treatment period. As these slight clinical signs were mostly transient and seen in isolated animals, they were considered to be incidental.
During the weekly detailed clinical observation, no significant changes or differences between the groups were found except statistically significantly higher or lower mean score for few parameters like salivation, spontaneous activity, animal sleeps, animal moving in cage and salivation on few occasions without showing any consistency in MD or HD group females and higher mean score for salivation in HD group males when compared with the controls. There was also a statistically significantly lower mean score for animal moving in cage in male high dose recovery group in week 5 and higher mean score for salivation in the female high dose recovery group in week 8. This statistically significant effect on detailed clinical examination was not considered to be treatment related.
Mortality:
no mortality observed
Description (incidence):
No mortality occurred in the control or any of the dose groups during the treatment period or recovery period of this study.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
In males sacrificed at the end of treatment period, a tendency towards a marginally lower mean body weight was observed in the HD group after the day 57 of treatment compared to the control group and the body weight was noted to be further slightly reduced until the end of the treatment period (up to 3%; not statistically significant). In males sacrificed at the end of the recovery period, during the treatment period and recovery period, mean body weight of the male HD recovery group remained slightly, lower compared to the control group (up to 8%; not statistically significant). In males sacrificed at the end of treatment period, body weight gain was lower from day 36 onwards in the MD group and day 8 onwards in the HD group until termination, without achieving statistical significance when compared to the controls. In males sacrificed at the end of the recovery period, statistically significantly lower group mean body weight gain was observed in the HD recovery group during day 43-50, 57-64 and statistically significantly higher group mean body weight gain was observed during day 78-85 and day 111-118 when compared with the controls.
In females, a tendency towards a marginally lower mean body weight was observed in the MD and HD groups from day/week 1 of treatment compared to the control group and the body weight was noted to be further slightly reduced until the end of the treatment period (up to 6.4%) without achieving statistical significance. In females sacrificed at the end of the recovery period, during the treatment period and recovery period, mean body weight of the female HD recovery group was statistically significantly lower on day 8 compared to the control group (up to 4.4%). However, on other days of the treatment period and all days of the recovery period, group mean body weight was comparable with the controls. In females sacrificed at the end of the treatment period, body weight gain was statistically significantly lower in LD and MD groups during day 22-29 when compared with the controls. Overall mean daily weight gain (Day 1-90) during the treatment period was slightly lower in the female HD group when compared to the controls. In females sacrificed at the end of the recovery period, statistically significantly lower body weight gain was observed in the HD recovery group during day 1-8 when compared with the controls.
As differences in body weight gain were marginal and values were within the normal range of variation for animals of this strain, effects on body weight and body weight gain were not considered as an adverse effect of the test item
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
The test item had no effect on food consumption in this study. Mean daily food intake of male and female animals was in the normal range of variation throughout the treatment and recovery periods of this study.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Description (incidence and severity):
No ophthalmologic findings were observed in any of the animals of this study.
Haematological findings:
no effects observed
Description (incidence and severity):
In males and females sacrificed at the end of the treatment period and recovery period, no test item-related effect of toxicological relevance was observed for haematology and coagulation parameters.
In males sacrificed at the end of the treatment period, statistical analysis of haematology and coagulation data revealed, statistically significantly lower HCT group mean values in LD and group mean reticulocytes values in HD group when compared with the controls. In males sacrificed at the end of the recovery period, statistically significantly lower WBC count was observed in HD recovery group when compared with the recovery controls.
In females sacrificed at the end of the treatment period, there was a marginally but statistically significantly lower mean value of mean corpuscular volume (MCV) and higher value of mean corpuscular haemoglobin concentration (MCHC) in the HD group compared with the controls. Statistical analysis also revealed a statistically significantly lower group mean basophil count in LD and MD groups without following a dose-related pattern when compared with the controls. In females sacrificed at the end of the recovery period, no statistically significant effect was observed for any haematology parameter when compared with the recovery controls.
As the respective haematology values were within the normal range of variation for animals of this strain and age, differences are not assumed to be biologically relevant even though they were statistically significant.
All haematological parameters and blood coagulation parameters in males and females sacrificed at the end of treatment and recovery periods were in the normal range of variation and no statistically significant or test item-related effects were observed.

Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
In males and females sacrificed at the end of the treatment period and recovery period, no test item-related effect of toxicological relevance was observed for clinical biochemistry parameters.
In males sacrificed at the end of treatment period, statistical analysis of clinical biochemistry data revealed, statistically significantly higher TBA group mean values in the HD group when compared with the controls. In males sacrificed at the end of the recovery period, marginally but statistically significantly higher ALAT was observed in the HD recovery group when compared with the controls.
In females sacrificed at the end of the treatment period, no statistically significant effect was observed for any of the clinical biochemistry parameters when compared with the controls. In females sacrificed at the end of the recovery period, statistically significantly lower group mean values for TP, Alb and Na was observed in the HD recovery group when compared with the recovery controls.
As the respective clinical biochemistry values were either within the normal range of variation for animals of this strain and age and/or no such effect was observed at the end of the treatment period, these differences are assumed not to be biologically relevant even though they were statistically significant.
All clinical biochemistry parameters in males and females sacrificed at the end of treatment and recovery period were in the normal range of variation and no statistically significant or test item-related effects were observed.
Urinalysis findings:
no effects observed
Description (incidence and severity):
The test item had no toxicologically relevant effect on urinary parameters analysed at the end of the treatment period and recovery period of this study.
Slightly higher leukocyte levels were found in the urine of the majority of male animals (all groups including controls) at the end of the treatment period, and in few control and HD males sacrificed at the end of the recovery period. In females, this finding was observed in a few animals of all groups (including controls) sacrificed at the end of treatment period and in only a few animals of control recovery group. No such finding was observed in HD recovery females sacrificed at the end of recovery period.
In males and females, all other urinary parameters were in the normal range of variation and no test item-related differences between the dose groups and control group were observed.
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
No relevant and test item related effects were observed in any of the parameters of the functional observation battery at the end of the treatment period and at the end of the recovery period.
Before initiation of treatment, there was a statistically significantly higher group mean score for head touch, ground righting reflex, air righting reflex in males treatment groups and head touch, fear, defecation and equilibrium reflex in MD and HD female groups when compared with the controls. As this effect was observed before initiation of the treatment, no toxicological significance can be attributed to these findings.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
The test item had no toxicologically relevant effect on organ weights determined at the end of the treatment period and recovery period of this study. All group mean and individual values for the male and female organs weights (absolute and relative to body weight and brain weight) from the test item treated groups were comparable with the control group.
No statistically significant effect on organs weights was observed in male and females sacrificed at the end of treatment period and recovery period except statistically significantly higher relative (to body weight) kidney weights in HD recovery males when compared with respective control group. This effect on kidney weight in HD male recovery animals was not considered to be test item related as no such effect was observed in males sacrificed at the end of the treatment period.
Gross pathological findings:
no effects observed
Description (incidence and severity):
Few specific gross pathological changes in the uterus were recorded for a few female animals. These findings were not considered to be treatment-related and assumed to be common background findings in this strain.
Complete dilatation of uterus was observed in 3 females of LD (61, 68, 69), 2 females of MD (73, 75) and 1 female of HD (81) group.
No gross pathological findings were observed in males sacrificed at the end of the treatment period and both male and females sacrificed at the end of the recovery period
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Under the conditions of this study, accumulation of α2-microglobulin in kidneys from several test item-treated males and from control animals was observed. In addition, the comparison between group 4 (treatment and recovery period) and group 1 animals (treatment and recovery period) revealed similar α2-microglobulin accumulation. Further, the α2-microglobulin accumulation noticed after immunohistochemistry using a specific anti-Alpha 2u Globulin antibody corresponded with the presence of hyaline inclusions that was recorded in hematoxylin and eosin stained kidney sections. However, no dose dependency was observed during the histopathological evaluation.
The accumulation of α2-microglobulin comprises a male-rat-specific syndrome without toxicological relevance in humans. In absence of further renal lesions, this change was considered non-adverse.
All remaining findings recorded were considered incidental or were within the range of spontaneous background alterations that may be recorded in Wistar rats at these ages.

Effect levels

open allclose all
Key result
Dose descriptor:
NOAEL
Effect level:
>= 100 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: No adverse effects observed.
Remarks on result:
other: See 'any other information on results incl. tables' and attachments for results tables
Key result
Dose descriptor:
NOAEL
Effect level:
>= 100 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: No adverse effects observed.
Remarks on result:
other: See 'any other information on results incl. tables' and attachments for results tables

Target system / organ toxicity

Critical effects observed:
no

Any other information on results incl. tables

Summary hematology

Table 1: Sex: male; phase: in life

 

 

C / M

LD / M

MD / M

HD / M

RBC

[M/µL] day 91

Mean

S.d.

N

Deviation Vs Control [%]

9.171a

0.501 10

8.867

0.387

9 -3.318

9.036

0.314

8 -1.469

9.296

0.379

10

1.363

HGB

[g/dL] day 91

Mean

S.d.

N

Deviation Vs Control [%]

15.84a

0.64 10

15.46

0.36

9 -2.43

15.55

0.59

8 -1.83

15.89

0.43

10

0.32

WBC

[K/uL] day 91

Mean

S.d.

N

Deviation Vs Control [%]

5.079k

1.543 10

4.720

0.974

9 -7.068

4.946

1.403

8 -2.614

5.604

1.437

10

10.337

HCT

[%] day 91

Mean

S.d.

N

Deviation Vs Control [%]

50.48ad

2.35 10

48.39*

1.43

9 -4.14

48.90

1.67

8 -3.13

50.55

1.41

10

0.14

MCV

[fl]

day 91

Mean

S.d.

N

Deviation Vs Control [%]

55.09k

2.06 10

54.63

1.56

9 -0.83

54.14

1.45

8 -1.73

54.41

1.15

10 -1.23

MCH

[pg] day 91

Mean

S.d.

N

Deviation Vs Control [%]

17.32a

0.72 10

17.48

0.68

9

0.91

17.21

0.50

8 -0.62

17.12

0.39

10 -1.15

MCHC

[g/dL] day 91

Mean

S.d.

N

Deviation Vs Control [%]

31.41a

0.52 10

31.93

0.72

9

1.67

31.84

0.46

8

1.36

31.45

0.59

10

0.13

RET

[%] day 91

Mean

S.d.

N

Deviation Vs Control [%]

1.522ad

0.125 10

1.599

0.232

9

5.052

1.406

0.197

8 -7.605

1.261**

0.149

10 -17.148

PLT

[K/uL] day 91

Mean

S.d.

N

Deviation Vs Control [%]

716.7a

90.9 10

712.0

78.5

9

-0.7

745.6

83.7

8

4.0

713.7

44.6

10

-0.4

LYM

[%] day 91

Mean

S.d.

N

Deviation Vs Control [%]

75.45a

6.79 10

75.36

7.64

9 -0.13

74.54

3.58

8 -1.21

78.72

3.18

10

4.33

NEUT

[%] day 91

Mean

S.d.

N

Deviation Vs Control [%]

21.20a

6.76 10

21.71

7.40

9

2.41

22.00

3.75

8

3.77

17.80

3.08

10 -16.04

EOS

[%] day 91

Mean

S.d.

N

Deviation Vs Control [%]

0.34k

0.20 10

0.33

0.17

9 -1.96

0.49

0.19

8

43.38

0.31

0.13

10 -8.82

MONO

[%] day 91

Mean

S.d.

N

Deviation Vs Control [%]

2.03a

0.49 10

1.68

0.45

9 -17.35

2.01

0.65

8 -0.86

2.07

0.57

10

1.97

BASO

[%] day 91

Mean

S.d.

N

Deviation Vs Control [%]

0.32k

0.11 10

0.31

0.09

9 -2.78

0.35

0.13

8

9.38

0.26

0.11

10 -18.75

LUC%

[%] day 91

Mean

S.d.

N

Deviation Vs Control [%]

0.66a

0.31 10

0.62

0.17

9 -5.72

0.60

0.15

8 -9.09

0.82

0.32

10

24.24

a=ANOVA; k=KRUSKALL-WALLIS; ad=ANOVA-DUNNETT; * = p < 0.05; ** = p < 0.01

Table 2: sex: females; phase: in life

 

 

C / F

LD / F

MD / F

HD / F

RBC

[M/µL] day 91

Mean

S.d.

N

Deviation Vs Control [%]

7.562k

0.395 10

7.313

0.428

10 -3.293

7.392

0.261

10 -2.248

7.487

0.632

9 -0.996

HGB

[g/dL] day 91

Mean

S.d.

N

Deviation Vs Control [%]

13.71a

0.63 10

13.28

0.73

10 -3.14

13.53

0.28

10 -1.31

13.46

1.13

9 -1.86

WBC

[K/uL] day 91

Mean

S.d.

N

Deviation Vs Control [%]

1.250a

0.941 10

0.813

0.516

10 -34.960

0.858

0.611

10 -31.360

1.281

0.686

9

2.489

HCT

[%] day 91

Mean

S.d.

N

Deviation Vs Control [%]

43.33a

2.29 10

41.30

2.28

10 -4.68

41.85

1.25

10 -3.42

41.10

3.62

9 -5.15

MCV

[fl]

day 91

Mean

S.d.

N

Deviation Vs Control [%]

57.34ad

1.72 10

56.48

1.37

10 -1.50

56.62

1.10

10 -1.26

54.90**

1.32

9 -4.26

MCH

[pg] day 91

Mean

S.d.

N

Deviation Vs Control [%]

18.13a

0.37 10

18.19

0.61

10

0.33

18.31

0.36

10

0.99

17.99

0.54

9 -0.78

MCHC

[g/dL] day 91

Mean

S.d.

N

Deviation Vs Control [%]

31.66ad

0.95 10

32.18

0.71

10

1.64

32.37

0.74

10

2.24

32.78**

0.48

9

3.53

RET

[%] day 91

Mean

S.d.

N

Deviation Vs Control [%]

1.954a

0.298 10

1.946

0.361

10 -0.409

2.011

0.222

10

2.917

1.924

0.286

9 -1.513

PLT

[K/uL] day 91

Mean

S.d.

N

Deviation Vs Control [%]

691.7a

62.5 10

691.4

75.9

10 0.0

661.7

108.4

10

-4.3

711.7

31.1

9

2.9

LYM

[%] day 91

Mean

S.d.

N

Deviation Vs Control [%]

73.64k

11.49 10

71.93

8.57

10 -2.32

66.48

14.60

10 -9.72

72.74

8.32

9 -1.22

NEUT

[%] day 91

Mean

S.d.

N

Deviation Vs Control [%]

21.51k

9.59 10

25.24

8.50

10

17.34

29.61

13.78

10

37.66

23.58

8.01

9

9.61

EOS

[%] day 91

Mean

S.d.

N

Deviation Vs Control [%]

1.56k

2.67 10

0.65

0.61

10 -58.33

1.22

1.08

10 -21.79

0.88

0.78

9 -43.73

MONO

[%] day 91

Mean

S.d.

N

Deviation Vs Control [%]

2.39a

0.77 10

1.75

0.75

10 -26.78

2.22

0.72

10 -7.11

2.04

0.83

9 -14.46

BASO

[%] day 91

Mean

S.d.

N

Deviation Vs Control [%]

0.38u

0.19 10

0.11**

0.12

10 -71.05

0.11**

0.17

10 -71.05

0.29

0.19

9 -23.98

LUC%

[%] day 91

Mean

S.d.

N

Deviation Vs Control [%]

0.53k

0.39 10

0.31

0.30

10 -41.51

0.35

0.21

10 -33.96

0.49

0.36

9 -7.76

k=KRUSKALL-WALLIS; a=ANOVA; ad=ANOVA-DUNNETT; ** = p < 0.01

Table 3: sex: males recovery group; phase: in life

 

 

Recovery C / M

Recovery HD / M

RBC

[M/µL] day 119

Mean

S.d.

N

Deviation Vs Control [%]

9.192a

0.262 4

8.524

1.010

5 -7.272

HGB

[g/dL] day 119

Mean

S.d.

N

Deviation Vs Control [%]

15.95s

0.26 4

14.84

1.69

5 -6.96

WBC

[K/uL] day 119

Mean

S.d.

N

Deviation Vs Control [%]

4.898ad

1.043 4

2.968*

0.776

5 -39.398

HCT

[%] day 119

Mean

S.d.

N

Deviation Vs Control [%]

48.90a

0.08 4

45.72

5.26

5 -6.50

MCV

[fl]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

53.25a

1.55 4

53.66

1.35

5

0.77

MCH

[pg] day 119

Mean

S.d.

N

Deviation Vs Control [%]

17.38a

0.28 4

17.42

0.44

5

0.26

MCHC

[g/dL] day 119

Mean

S.d.

N

Deviation Vs Control [%]

32.60a

0.50 4

32.50

0.51

5 -0.31

RET

[%] day 119

Mean

S.d.

N

Deviation Vs Control [%]

1.712a

0.240 4

1.784

0.280

5

4.175

PLT

[K/uL] day 119

Mean

S.d.

N

Deviation Vs Control [%]

765.8s

73.8 4

617.8

327.0

5 -19.3

LYM

[%] day 119

Mean

S.d.

N

Deviation Vs Control [%]

77.90a

6.31 4

72.38

3.22

5 -7.09

NEUT

[%] day 119

Mean

S.d.

N

Deviation Vs Control [%]

18.30a

6.42 4

24.32

3.41

5

32.90

EOS

[%] day 119

Mean

S.d.

N

Deviation Vs Control [%]

0.48a

0.10 4

0.42

0.16

5 -11.58

MONO

[%] day 119

Mean

S.d.

N

Deviation Vs Control [%]

2.42a

0.60 4

2.10

0.41

5 -13.40

BASO

[%] day 119

Mean

S.d.

N

Deviation Vs Control [%]

0.35s

0.06 4

0.32

0.08

5 -8.57

LUC%

[%] day 119

Mean

S.d.

N

Deviation Vs Control [%]

0.60a

0.37 4

0.42

0.19

5 -30.00

a=ANOVA; s=DUNN; ad=ANOVA-DUNNETT; * = p < 0.05

Table 4: sex: females recovery group; phase: in life

 

 

Recovery C / F

Recovery HD / F

RBC

[M/µL] day 119

Mean

S.d.

N

Deviation Vs Control [%]

6.648s

2.947 5

8.268

0.226

5

24.368

HGB

[g/dL] day 119

Mean

S.d.

N

Deviation Vs Control [%]

14.46a

1.06 5

14.82

0.24

5

2.49

WBC

[K/uL] day 119

Mean

S.d.

N

Deviation Vs Control [%]

1.236a

0.714 5

1.520

0.475

5

22.977

HCT

[%] day 119

Mean

S.d.

N

Deviation Vs Control [%]

44.98a

3.68 5

46.26

0.73

5

2.85

MCV

[fl]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

57.20a

1.24 5

55.96

0.97

5 -2.17

MCH

[pg] day 119

Mean

S.d.

N

Deviation Vs Control [%]

18.40a

0.64 5

17.94

0.40

5 -2.50

MCHC

[g/dL] day 119

Mean

S.d.

N

Deviation Vs Control [%]

32.20a

0.50 5

32.10

0.28

5 -0.31

RET

[%] day 119

Mean

S.d.

N

Deviation Vs Control [%]

1.706a

0.170 5

1.830

0.260

5

7.268

PLT

[K/uL] day 119

Mean

S.d.

N

Deviation Vs Control [%]

723.4a

100.3 5

728.0

82.3

5

0.6

LYM

[%] day 119

Mean

S.d.

N

Deviation Vs Control [%]

76.22a

4.60 5

76.04

8.52

5 -0.24

NEUT

[%] day 119

Mean

S.d.

N

Deviation Vs Control [%]

19.46a

4.02 5

20.48

7.62

5

5.24

EOS

[%] day 119

Mean

S.d.

N

Deviation Vs Control [%]

1.38a

0.78 5

0.82

0.61

5 -40.58

MONO

[%] day 119

Mean

S.d.

N

Deviation Vs Control [%]

2.32a

1.69 5

2.02

1.05

5 -12.93

BASO

[%] day 119

Mean

S.d.

N

Deviation Vs Control [%]

0.26a

0.21 5

0.32

0.23

5

23.08

LUC%

[%] day 119

Mean

S.d.

N

Deviation Vs Control [%]

0.38a

0.23 5

0.30

0.21

5 -21.05

s=DUNN; a=ANOVA

Summary coagulation

Table 1: In life, males

 

 

C / M

LD / M

MD / M

HD / M

PT

[sec] day 91

Mean

S.d.

N

Deviation Vs Control [%]

20.25k

1.25 10

19.31

0.74

9 -4.64

20.67

1.38

9

2.06

20.42

1.64

10

0.84

APTT

[sec] day 91

Mean

S.d.

N

Deviation Vs Control [%]

11.47k

3.45 10

9.97

2.71

9 -13.11

11.33

4.85

10 -1.22

9.70

1.60

10 -15.43

k=KRUSKALL-WALLIS

Table 2: In life, females

 

 

C / F

LD / F

MD / F

HD / F

PT

[sec] day 91

Mean

S.d.

N

Deviation Vs Control [%]

21.24a

1.74 10

21.50

1.67

9

1.22

21.53

2.08

10

1.37

22.03

2.14

10

3.72

APTT

[sec] day 91

Mean

S.d.

N

Deviation Vs Control [%]

9.37a

2.09 10

9.27

2.59

9 -1.10

9.57

1.80

10

2.13

9.86

1.68

10

5.23

a=ANOVA

Table 3: In life, recovery males

 

 

Recovery C / M

Recovery HD / M

PT

[sec] day 119

Mean

S.d.

N

Deviation Vs Control [%]

19.18a

1.02 5

20.44

1.50

5

6.57

APTT

[sec] day 119

Mean

S.d.

N

Deviation Vs Control [%]

12.64a

1.82 5

13.50

4.23

5

6.80

a=ANOVA

Table 4: In life, recovery females

 

 

Recovery C / F

Recovery HD / F

PT

[sec] day 119

Mean

S.d.

N

Deviation Vs Control [%]

19.62a

1.43 4

20.10

1.71

5

2.42

APTT

[sec] day 119

Mean

S.d.

N

Deviation Vs Control [%]

12.22a

3.68 5

12.84

0.99

5

5.07

a=ANOVA

Summary clinical biochemistry

Table 1: In life males

 

 

C / M

LD / M

MD / M

HD / M

ALAT

[U/l]

day 91

Mean

S.d.

N

Deviation Vs Control [%]

33.80a

3.81 10

35.18

7.63

10

4.08

40.61

9.71

10

20.15

40.16

5.52

10

18.82

ASAT

[U/l]

day 91

Mean

S.d.

N

Deviation Vs Control [%]

104.59k

20.02 10

99.12

25.07

10 -5.23

108.73

31.48

10

3.96

94.93

10.10

10 -9.24

AP

[U/l]

day 91

Mean

S.d.

N

Deviation Vs Control [%]

136.081u

53.601 10

93.862

17.596

10 -31.025

124.415

32.155

10 -8.573

173.184

95.364

10

27.265

Crea

[µmol/L] day 91

Mean

S.d.

N

Deviation Vs Control [%]

25.5a

2.5 10

25.5

3.7

10 0.0

27.8

6.9

10 9.0

24.9

3.9

10

-2.4

TP

[G/l]

day 91

Mean

S.d.

N

Deviation Vs Control [%]

55.18ad

1.36 10

56.74

1.61

10

2.83

55.13

1.44

10 -0.09

54.32

1.37

10 -1.56

Alb

[G/l]

day 91

Mean

S.d.

N

Deviation Vs Control [%]

30.663a

0.823 10

30.986

1.125

10

1.053

30.213

0.820

10 -1.468

30.467

0.609

10 -0.639

urea

[mmol/l] day 91

Mean

S.d.

N

Deviation Vs Control [%]

5.814a

0.923 10

5.129

0.686

10 -11.782

5.641

0.719

10 -2.976

5.766

0.911

10 -0.826

TBIL

[µmol/L] day 91

Mean

S.d.

N

Deviation Vs Control [%]

2.02k

0.34 10

2.26

0.96

10

11.88

2.21

0.50

10

9.41

2.19

0.31

10

8.42

TBA

[µmol/L] day 91

Mean

S.d.

N

Deviation Vs Control [%]

34.497ad

5.967 10

31.342

10.317

10 -9.146

38.267

10.500

10

10.928

50.048**

11.752

10

45.079

Chol

[mmol/l] day 91

Mean

S.d.

N

Deviation Vs Control [%]

1.624a

0.319 10

1.900

0.299

10

16.995

1.672

0.266

10

2.956

1.641

0.140

10

1.047

Gluc

[mmol/l] day 91

Mean

S.d.

N

Deviation Vs Control [%]

9.540a

1.089 10

10.520

2.400

10

10.273

8.587

1.861

10 -9.990

9.615

1.044

10

0.786

Na

[mmol/l] day 91

Mean

S.d.

N

Deviation Vs Control [%]

141.8k

0.9 10

142.2

1.4

10 0.3

140.9

2.1

10

-0.6

141.4

1.6

10

-0.3

K

[mmol/l] day 91

Mean

S.d.

N

Deviation Vs Control [%]

4.344k

0.668 10

4.372

1.011

10

0.645

4.629

1.352

10

6.561

4.354

0.628

10

0.230

a=ANOVA; k=KRUSKALL-WALLIS; u=KRUSKALL-WALLIS-DUNN; ad=ANOVA-DUNNETT; ** = p < 0.01

Table 2: In life females

 

 

C / F

LD / F

MD / F

HD / F

ALAT

[U/l]

day 91

Mean

S.d.

N

Deviation Vs Control [%]

29.85a

12.15 10

29.47

6.08

10 -1.27

27.47

5.25

10 -7.97

26.12

5.18

10 -12.50

ASAT

[U/l]

day 91

Mean

S.d.

N

Deviation Vs Control [%]

87.13a

18.56 10

81.28

9.59

10 -6.71

80.40

12.15

10 -7.72

78.51

20.37

10 -9.89

AP

[U/l]

day 91

Mean

S.d.

N

Deviation Vs Control [%]

59.410k

27.714 10

45.603

16.830

10 -23.240

58.686

22.900

10 -1.219

61.329

24.534

10

3.230

Crea

[µmol/L] day 91

Mean

S.d.

N

Deviation Vs Control [%]

32.5a

4.9 10

31.2

4.6

9

-3.9

28.7

4.7

10 -11.7

29.4

2.4

10

-9.5

TP

[G/l]

day 91

Mean

S.d.

N

Deviation Vs Control [%]

66.12k

4.28 10

63.66

3.63

9 -3.73

65.84

4.15

10 -0.42

63.65

3.20

10 -3.74

Alb

[G/l]

day 91

Mean

S.d.

N

Deviation Vs Control [%]

37.573a

2.285 10

36.454

2.175

10 -2.978

37.218

3.008

10 -0.945

35.764

2.062

10 -4.815

urea

[mmol/l] day 91

Mean

S.d.

N

Deviation Vs Control [%]

6.571k

0.766 10

6.090

0.596

9 -7.320

6.393

0.694

10 -2.709

6.489

1.173

10 -1.248

TBIL

[µmol/L] day 91

Mean

S.d.

N

Deviation Vs Control [%]

2.32a

0.32 10

2.29

0.31

9 -1.34

2.26

0.37

10 -2.59

2.37

0.61

10

2.16

TBA

[µmol/L] day 91

Mean

S.d.

N

Deviation Vs Control [%]

31.261a

32.784 10

22.624

9.586

9 -27.627

18.381

6.625

10 -41.202

34.996

25.785

10

11.948

Chol

[mmol/l] day 91

Mean

S.d.

N

Deviation Vs Control [%]

1.504a

0.267 10

1.488

0.353

10 -1.064

1.417

0.216

10 -5.785

1.520

0.184

10

1.064

Gluc

[mmol/l] day 91

Mean

S.d.

N

Deviation Vs Control [%]

7.504a

1.324 10

7.589

1.469

9

1.131

7.130

1.584

10 -4.984

7.332

1.127

10 -2.292

Na

[mmol/l] day 91

Mean

S.d.

N

Deviation Vs Control [%]

138.6k

2.8 10

139.0

1.9

10 0.3

140.5

2.2

10 1.4

139.7

2.6

10 0.8

K

[mmol/l] day 91

Mean

S.d.

N

Deviation Vs Control [%]

3.797k

0.520 9

3.733

0.380

10 -1.677

3.577

0.327

10 -5.786

3.692

0.204

10 -2.757

a=ANOVA; k=KRUSKALL-WALLIS

Table 3: In life recovery males

 

 

Recovery C / M

Recovery HD / M

ALAT

[U/l]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

25.12ad

0.53 5

29.18*

3.69

5

16.16

ASAT

[U/l]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

83.30a

16.08 5

93.72

13.59

5

12.51

AP

[U/l]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

104.536s

19.014 5

96.296

36.405

5 -7.882

Crea

[µmol/L] day 119

Mean

S.d.

N

Deviation Vs Control [%]

25.0a

3.4 5

23.0

2.1

5

-8.0

TP

[G/l]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

53.26s

4.96 5

57.08

4.18

5

7.17

Alb

[G/l]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

28.612a

2.816 5

31.230

2.486

5

9.150

urea

[mmol/l] day 119

Mean

S.d.

N

Deviation Vs Control [%]

6.834a

0.557 5

6.408

0.292

5 -6.234

TBIL

[µmol/L] day 119

Mean

S.d.

N

Deviation Vs Control [%]

1.96s

0.31 5

2.08

0.24

5

6.12

TBA

[µmol/L] day 119

Mean

S.d.

N

Deviation Vs Control [%]

25.960a

5.055 5

25.348

5.567

5 -2.357

Chol

[mmol/l] day 119

Mean

S.d.

N

Deviation Vs Control [%]

1.592a

0.232 5

1.650

0.211

5

3.643

Gluc

[mmol/l] day 119

Mean

S.d.

N

Deviation Vs Control [%]

11.924a

2.191 5

9.442

0.997

5 -20.815

Na

[mmol/l] day 119

Mean

S.d.

N

Deviation Vs Control [%]

128.2a

9.4 5

133.6

10.2

5

4.2

K

[mmol/l] day 119

Mean

S.d.

N

Deviation Vs Control [%]

3.822a

0.231 5

3.626

0.328

5 -5.128

 

 

Recovery C / M

Recovery HD / M

ALAT

[U/l]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

25.12ad

0.53 5

29.18*

3.69

5

16.16

ASAT

[U/l]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

83.30a

16.08 5

93.72

13.59

5

12.51

AP

[U/l]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

104.536s

19.014 5

96.296

36.405

5 -7.882

Crea

[µmol/L] day 119

Mean

S.d.

N

Deviation Vs Control [%]

25.0a

3.4 5

23.0

2.1

5

-8.0

TP

[G/l]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

53.26s

4.96 5

57.08

4.18

5

7.17

Alb

[G/l]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

28.612a

2.816 5

31.230

2.486

5

9.150

urea

[mmol/l] day 119

Mean

S.d.

N

Deviation Vs Control [%]

6.834a

0.557 5

6.408

0.292

5 -6.234

TBIL

[µmol/L] day 119

Mean

S.d.

N

Deviation Vs Control [%]

1.96s

0.31 5

2.08

0.24

5

6.12

TBA

[µmol/L] day 119

Mean

S.d.

N

Deviation Vs Control [%]

25.960a

5.055 5

25.348

5.567

5 -2.357

Chol

[mmol/l] day 119

Mean

S.d.

N

Deviation Vs Control [%]

1.592a

0.232 5

1.650

0.211

5

3.643

Gluc

[mmol/l] day 119

Mean

S.d.

N

Deviation Vs Control [%]

11.924a

2.191 5

9.442

0.997

5 -20.815

Na

[mmol/l] day 119

Mean

S.d.

N

Deviation Vs Control [%]

128.2a

9.4 5

133.6

10.2

5

4.2

K

[mmol/l] day 119

Mean

S.d.

N

Deviation Vs Control [%]

3.822a

0.231 5

3.626

0.328

5 -5.128

ad=ANOVA-DUNNETT; * = p < 0.05; a=ANOVA; s=DUNN

Table 4: In life recovery females

 

 

Recovery C / F

Recovery HD / F

ALAT

[U/l]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

32.70a

5.55 5

25.44

4.39

5 -22.20

ASAT

[U/l]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

101.42a

19.86 5

94.44

17.33

5 -6.88

AP

[U/l]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

54.378a

18.558 5

48.572

21.162

5 -10.677

Crea

[µmol/L] day 119

Mean

S.d.

N

Deviation Vs Control [%]

29.0s

1.9 5

50.4

49.6

5

73.8

TP

[G/l]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

62.94s

4.07 5

56.08*

5.30

5 -10.90

Alb

[G/l]

day 119

Mean

S.d.

N

Deviation Vs Control [%]

36.328s

2.499 5

31.626**

2.152

5 -12.943

urea

[mmol/l] day 119

Mean

S.d.

N

Deviation Vs Control [%]

7.332s

0.424 5

10.230

7.603

5

39.525

TBIL

[µmol/L] day 119

Mean

S.d.

N

Deviation Vs Control [%]

3.04a

0.47 5

3.88

2.23

5

27.63

TBA

[µmol/L] day 119

Mean

S.d.

N

Deviation Vs Control [%]

33.438a

20.590 5

18.504

15.399

5 -44.662

Chol

[mmol/l] day 119

Mean

S.d.

N

Deviation Vs Control [%]

1.342a

0.318 5

1.324

0.164

5 -1.341

Gluc

[mmol/l] day 119

Mean

S.d.

N

Deviation Vs Control [%]

6.636a

1.632 5

5.808

0.860

5 -12.477

Na

[mmol/l] day 119

Mean

S.d.

N

Deviation Vs Control [%]

138.0s

2.8 5

125.4*

10.3

5

-9.1

K

[mmol/l] day 119

Mean

S.d.

N

Deviation Vs Control [%]

3.494s

0.201 5

4.048

1.674

5

15.856

a=ANOVA; s=DUNN; * = p < 0.05; ** = p < 0.01

Summry gross pathology

Table 1: Males

Sex                                                             

               Dose Group         

M

C / M

M

LD / M

M      

MD / M

M

HD / M

BRAIN

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SPINAL CORD, CERVIC.

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SPINAL CORD, THORAC.

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SPINAL CORD, LUMBAR

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SCIATIC NERVE, LEFT

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

HEART

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

AORTA

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

TRACHEA

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

LUNG

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

TONGUE

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

ESOPHAGUS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

STOMACH

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

DUODENUM

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

JEJUNUM

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

ILEUM

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

PEYER'S PATCHES

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

CECUM

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

COLON

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

RECTUM

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

LIVER

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

PANCREAS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

KIDNEYS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

URETERS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

URINARY BLADDER

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

TESTES

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

EPIDIDYMIDES

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

PROSTATE GLAND

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

COAGULATING GLANDS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SEMINAL VESICLES

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

PITUITARY GLAND

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

THYROID GLAND

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

PARATHYROID GLANDS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

ADRENAL GLANDS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SPLEEN

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

BONE MARROW

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

THYMUS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

AXILLARY LYMPH NODES

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

MESENT. LYMPH NODE

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

MANDIB.LYMPH NODES

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SALIVARY GLANDS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

HARDERIAN GLANDS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

MAMMARY GLAND

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SKIN/SUBCUTIS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SKELETAL MUSCLE

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

EYES

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

FEMUR with KNEE JOIN

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

STERNUM

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

Table 2: Females

Sex                                                             

               Dose Group         

F

C/F

F

LD /F

F       

MD/F

F

HD /F

BRAIN

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SPINAL CORD, CERVIC.

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SPINAL CORD, THORAC.

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SPINAL CORD, LUMBAR

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SCIATIC NERVE, LEFT

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

HEART

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

AORTA

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

TRACHEA

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

LUNG

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

TONGUE

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

ESOPHAGUS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

STOMACH

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

DUODENUM

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

JEJUNUM

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

ILEUM

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

PEYER'S PATCHES

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

CECUM

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

COLON

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

RECTUM

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

LIVER

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

PANCREAS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

KIDNEYS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

URETERS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

URINARY BLADDER

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

OVARIES

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

OVIDUCTS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

UTERUS(dilatation complete)

 

 

 

 

Total affected /examined:

0/10

3/10

2/10

1/10

CERVIX

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

VAGINA

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

PITUITARY GLAND

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

THYROID GLAND

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

PARATHYROID GLANDS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

ADRENAL GLANDS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SPLEEN

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

BONE MARROW

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

THYMUS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

AXILLARY LYMPH NODES

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

MESENT. LYMPH NODE

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

MANDIB.LYMPH NODES

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SALIVARY GLANDS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

HARDERIAN GLANDS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

MAMMARY GLAND

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SKIN/SUBCUTIS

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

SKELETAL MUSCLE

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

EYES

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

FEMUR with KNEE JOIN

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

STERNUM

 

 

 

 

Total affected /examined:

0/10

0/10

0/10

0/10

Table 3: Recovery males and recovery females

Sex                                                             

               Dose Group         

M

RECOVERY C/M

M

RECOVERY HD/M

F        

RECOVERY C/F        

F

RECOVERY HD/F

BRAIN

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

SPINAL CORD, CERVIC.

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

SPINAL CORD, THORAC.

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

SPINAL CORD, LUMBAR

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

SCIATIC NERVE, LEFT

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

HEART

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

AORTA

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

TRACHEA

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

LUNG

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

TONGUE

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

ESOPHAGUS

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

STOMACH

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

DUODENUM

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

JEJUNUM

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

ILEUM

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

PEYER'S PATCHES

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

CECUM

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

COLON

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

RECTUM

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

LIVER

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

PANCREAS

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

KIDNEYS

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

URETERS

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

URINARY BLADDER

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

TESTES

 

 

 

 

Total affected /examined:

0/5

0/5

 

 

EPIDIDYMIDES

 

 

 

 

Total affected /examined:

0/5

0/5

 

 

PROSTATE GLAND

 

 

 

 

Total affected /examined:

0/5

0/5

 

 

COAGULATING GLANDS

 

 

 

 

Total affected /examined:

0/5

0/5

 

 

SEMINAL VESICLES

 

 

 

 

Total affected /examined:

0/5

0/5

 

 

OVARIES

 

 

 

 

Total affected /examined:

 

 

0/5

0/5

OVIDUCTS

 

 

 

 

Total affected /examined:

 

 

0/5

0/5

UTERUS(dilatation complete)

 

 

 

 

Total affected /examined:

 

 

0/5

0/5

CERVIX

 

 

 

 

Total affected /examined:

 

 

0/5

0/5

VAGINA

 

 

 

 

Total affected /examined:

 

 

0/5

0/5

PITUITARY GLAND

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

THYROID GLAND

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

PARATHYROID GLANDS

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

ADRENAL GLANDS

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

SPLEEN

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

BONE MARROW

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

THYMUS

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

AXILLARY LYMPH NODES

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

MESENT. LYMPH NODE

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

MANDIB.LYMPH NODES

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

SALIVARY GLANDS

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

HARDERIAN GLANDS

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

MAMMARY GLAND

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

SKIN/SUBCUTIS

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

SKELETAL MUSCLE

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

EYES

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

FEMUR with KNEE JOIN

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

STERNUM

 

 

 

 

Total affected /examined:

0/5

0/5

0/5

0/5

Microscopic findings

Table 1: Incidence and Mean Severity of Main Findings in Kidneys

Treatment period

 

 

 

 

Dose (mg/kg)

1

2

3

4

Finding / Groups

 

 

 

 

Total Animals/Sex

Affected / Mean Severity

10 M

10 M

10 M

 

10 M

Hyaline inclusions

10/1.3

9/1.6

10/1.7

10/1.6

Recovery period

 

 

 

 

Total Animals/Sex

Affected / Mean Severity

5 M

 

 

5 M

Hyaline inclusions

4/1.3

 

 

5/1.8

Table 2: Treatment groups: Summary of column statistics on relative positive area for α2-microglobulin

Group

1

2

3

4

Number of values

20

20

20

20

Minimum

0.0

0.0300

0.5500

0.2900

25% Percentile

0.2525

1.510

2.218

0.8600

Median

0.6000

2.435

4.245

1.275

75% Percentile

2.258

5.200

6.063

2.523

Maximum

4.840

6.780

7.840

2.870

Mean

1.283

2.994

4.239

1.517

Std. Deviation

1.459

2.172

2.181

0.8840

Std. Error of Mean

0.3262

0.4857

0.4877

0.1977

Lower 95% CI of mean

0.6003

1.977

3.218

1.103

Upper 95% CI of mean

1.966

4.011

5.259

1.931

Shapiro-Wilk normality test

W

0.7939

0.9060

0.9581

0.9091

P value

0.0007

0.0536

0.5070

0.0614

Passed normality test (alpha=0.05)?

No

Yes

Yes

Yes

P value summary

***

ns

ns

ns

Sum

25.66

59.88

84.77

30.34

Table 3: Treatment period - Single animal/kidney values of positive area in µm2

Group

Animal No.

Kidney 1

Kidney 2

 

Control (1)

1

2,54

3,48

Control (1)

2

0,56

1,41

Control (1)

3

0,72

0,89

Control (1)

4

0,31

0,2

Control (1)

5

0,25

0,63

Control (1)

6

0,49

1,35

Control (1)

7

0,07

0,1

Control (1)

8

0

0,26

Control (1)

9

0,57

3,14

Control (1)

10

3,85

4,84

Low dose (2)

11

1,51

0,61

Low dose (2)

12

5,38

5,11

Low dose (2)

13

2,36

2,61

Low dose (2)

14

4,7

5,69

Low dose (2)

15

1,83

5,23

Low dose (2)

16

1,69

2,51

Low dose (2)

17

0,12

0,03

Low dose (2)

18

1,22

2,72

Low dose (2)

19

1,57

1,51

Low dose (2)

20

6,7

6,78

Medium dose (3)

21

7,23

7,84

Medium dose (3)

22

5,5

5,74

Medium dose (3)

23

4,52

3,28

Medium dose (3)

24

2,33

5,49

Medium dose (3)

25

6,88

6,67

Medium dose (3)

26

1,84

3,87

Medium dose (3)

27

3,97

6,17

Medium dose (3)

28

1,35

0,55

Medium dose (3)

29

1,71

2,18

Medium dose (3)

30

2,62

5,03

High dose (4)

31

0,54

0,89

High dose (4)

32

2,04

2,69

High dose (4)

33

0,29

1,26

High dose (4)

34

0,85

1,08

High dose (4)

35

1,29

1,84

High dose (4)

36

0,33

1,21

High dose (4)

37

1,36

2,87

High dose (4)

38

0,54

2,83

High dose (4)

39

2,2

2,69

High dose (4)

40

0,91

2,63

Table 4: Recovery groups: Summary of column statistics on relative positive area for α2-microglobulin

Groups

1

4

Number of values

10

10

Minimum

0.3800

2.230

25% Percentile

1.743

2.945

Median

2.115

3.560

75% Percentile

3.933

5.173

Maximum

5.920

6.870

Mean

2.700

4.045

Std. Deviation

1.607

1.470

Std. Error of Mean

0.5082

0.4650

Lower 95% CI of mean

1.550

2.993

Upper 95% CI of mean

3.850

5.097

Shapiro-Wilk normality test

W

0.9410

0.9309

P value

0.5643

0.4569

Passed normality test (alpha=0.05)?

Yes

Yes

P value summary

ns

ns

Sum

27.00

40.45

Table 5: Treatment period - Single animal/kidney values of positive area in µm2

Group

Animal No.

Kidney 1

Kidney 2

 

Control (1)

41

0,38

2,02

Control (1)

42

2,1

1,45

Control (1)

43

3,79

4,36

Control (1)

44

2,13

1,84

Control (1)

45

3,01

5,92

High dose (4)

46

3,32

3,8

High dose (4)

47

4,67

6,87

High dose (4)

48

2,81

2,23

High dose (4)

49

2,99

5

High dose (4)

50

3,07

5,69

See attached documents for body and organ weights, pathology and histopatholagy report.

Applicant's summary and conclusion

Conclusions:
In the 90-day repeated dose oral toxicity study with tri(isopropyl)silyl acrylate (CAS 157859-20-6), conducted according to OECD Test Guideline 408 and in compliance with GLP, the reported NOAEL for specific target organ toxicity was greater than 100 mg/kg bw/day (the highest dose tested) based on no test item related effect observed on mortality, clinical signs, body weight development, food consumption, functional observation battery, weekly detailed clinical observations, haematology and blood coagulations, clinical biochemistry, urinalysis, gross pathological findings, organ weight and histopathology.