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Registration Dossier
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EC number: 278-770-4 | CAS number: 77804-81-0
Repeated dose toxicity: oral
Administrative data
- Endpoint:
- sub-chronic toxicity: oral
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2021
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 021
- Report date:
- 2021
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
- Deviations:
- no
- GLP compliance:
- yes
- Limit test:
- no
Test material
- Reference substance name:
- 2-[(4-chloro-2-nitrophenyl)azo]-N-(2,3-dihydro-2-oxo-1H-benzimidazol-5-yl)-3-oxobutyramide
- EC Number:
- 235-462-4
- EC Name:
- 2-[(4-chloro-2-nitrophenyl)azo]-N-(2,3-dihydro-2-oxo-1H-benzimidazol-5-yl)-3-oxobutyramide
- Cas Number:
- 12236-62-3
- Molecular formula:
- C17H13ClN6O5
- IUPAC Name:
- 2-[(4-chloro-2-nitrophenyl)diazenyl]-3-oxo-N-(2-oxo-2,3-dihydro-1H-benzimidazol-5-yl)butanamide
- Test material form:
- solid: nanoform
Constituent 1
Test animals
- Species:
- rat
- Strain:
- Wistar
- Details on species / strain selection:
- Rat is the standard laboratory rodent species used for toxicity assessment and recommended by various regulatory authorities.
The Wistar rat was selected due to the large amount of background data available for this strain. - Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- Hylasco Biotechnology Pvt. Ltd., Plot 4B, AKP, Turkapally Village, Shameerpet Mandal, RR Dist, Telangana 500078
- Females (if applicable) nulliparous and non-pregnant: [yes]
- Age at study initiation: 6 weeks
Body weight range at the start of treatment: Males: 189.68 to 249.91 g & Females : 156.56 to 192.12 g
At the commencement of the treatment, the weight variation of rats used did not exceed ± 20 % of the mean body weight in each sex and group.
Conditions: Rats were housed in an environment controlled room. The temperature maintained during the experiment was between 20 to 24°C and relative humidity was between 49 to 68%. The photoperiod was 12 hours light and 12 hours dark cycle. Adequate fresh air supply of 12-15 air changes/hour was maintained in the experimental room. The maximum and minimum temperature in the experimental room was recorded once daily. The relative humidity in the experimental room was calculated daily from dry and wet bulb temperature recordings.
Housing: Two rats of same sex were housed per cage in sterilized standard polysulfone cages (Size: L 425 x B 266 x H 185 mm), with stainless steel top grill having facilities for pelleted food and drinking water in polycarbonate bottles with stainless steel sipper tubes. The last animal in recovery group of each sex was housed individually. Polycarbonate rat huts were provided to the animals as environmental enrichment objects and changed along with cage at least once a week. During the experimental period, animals were housed in a single experimental room of barrier area.
Bedding: Steam sterilized corn cob was used as bedding and changed along with the cage atleast twice a week.
Diet: Altromin Rat/Mice Maintenance diets manufactured by Altromin Spezialfutter GmbH & Co. KG, Im Seelenkamp 20, 32791 Lage, Germany, was provided ad libitum.
Water: Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cum-purifier manufactured by Eureka Forbes Ltd., Mumbai 400 001,India, was provided ad libitum to rats in polycarbonate bottles with stainless steel sipper tubes.
Administration / exposure
- Route of administration:
- oral: gavage
- Details on route of administration:
- The dose formulations were administered orally by gavage to specific group of rats once daily at approximately the same time (± 3 hours) each day for a period of 90 consecutive days. Similarly, the vehicle was administered to rats in vehicle control/vehicle control recovery group once daily orally for 90 consecutive days.
The vehicle or the dose formulations were not administered to recovery groups for 28 days following the 90-day treatment period. The dose formulation and the vehicle were administered at an equivolume of 10 mL/kg/day. The dose volume was calculated for individual animals on the first day of treatment and was adjusted according to the most recent body weights recorded during the treatment period - Vehicle:
- CMC (carboxymethyl cellulose)
- Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- For homogeneity and active ingredient (a.i.) concentration analysis, prepared formulation samples were sampled in duplicate sets on Day 1 and during 2nd (Day 34) and 3rd (Day 63) month of the treatment period and analysed in-house. For each set, duplicate samples from top, middle and bottom layers were drawn from each preparation and in case of control, duplicate samples were drawn from middle layer.
The analysis was done as per the method validated under Eurofins Advinus Study No.: G19462. One set of samples was analyzed for concentration. The back up samples were discarded as analysis results of the first set of samples were within the acceptable limits. Formulations were considered acceptable as overall mean results of all the layers and mean of each layer were within ± 15.0 % of the claimed concentration and relative standard deviation (% RSD) was less than 10.0 %. - Duration of treatment / exposure:
- 90 Days
- Frequency of treatment:
- Daily
Doses / concentrationsopen allclose all
- Dose / conc.:
- 111 mg/kg bw/day (nominal)
- Dose / conc.:
- 333 mg/kg bw/day (nominal)
- Dose / conc.:
- 1 000 mg/kg bw/day (nominal)
- No. of animals per sex per dose:
- 10
- Control animals:
- yes
- Details on study design:
- No. of groups : 6
Vehicle control (G1)
Low dose (G2)
Mid dose (G3)
High dose (G4)
Vehicle control recovery (G1R)
High dose recovery (G4R)
No. of rats/group: Main groups: 10 males + 10 females
Recovery groups: 5 males + 5 females
Total = 100 (50 males + 50 females)
Examinations
- Observations and examinations performed and frequency:
- Observations and examinations performed and frequency
Morbidity and Mortality: All rats were observed for morbidity and mortalities twice daily i.e., once in the morning and once in the afternoon except during holidays wherein the observation was done once daily as there were no clinical signs observed. Clinical Signs: Each rat was observed for checking general clinical signs twice once daily during treatment period once daily during the recovery period. On the days of scheduled detailed clinical examination, clinical signs were included as a part of detailed clinical observations except on Day 1 wherein detailed clinical examination was done prior to the treatment and observations for general clinical signs was done after dosing the animals.
Detailed Clinical Examination: Detailed clinical examination was done prior to the test item administration on Day 1 and at weekly intervals thereafter (± 2 days) during treatment period. During detailed clinical examination, all rats were observed for changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions and autonomic activity (e.g. lacrimation, piloerection, pupil size, unusual respiratory pattern), changes in gait, posture and response to handling as well as the presen ce of clonic or tonic movements, stereotypies (e.g., excessive grooming, repetitive circling or bizarre behaviour (e.g. self-mutilation, walking backwards). On the days of detailed clinical examination, observation for general clinical signs (first post-dose) was not performed except on Day 1. Ophthalmological Examination: Ophthalmological examination of all animals was performed with an ophthalmoscope prior to start of the treatment, at the end of the treatment period for main groups (Day 84) and at the end of recovery period (Day 117)for recovery groups. Before examination, my driasis was induced using a 1 % solution of Tropicamide.
Functional Observation Battery Tests (FOB)
The following neurological examination was performed during the 12th week (Day 84) of treatment period for main groups and towards the end of recovery period (Day 117) for recovery groups.
Home Cage Observations: Each rat was observed in the home cage for posture and for presence or absence of abnormal vocalizations, tremors and convulsions.
Observations during Removal of Animal from Home Cage and Handling: The objective of this phase of neurological examination was to observe the subject’s response to handling and to conduct other procedures of the FOB that can best be performed when the rat is being held. Each rat was observed for the following examinations:
ease of removal from home cage
handling reactivity
palpebral closure
eye examination
piloerection
lacrimation
salivation
skin/fur examination
perineum wetness
respiration
muscle tone and
extensor thrust response
The observations were recorded using scores/ranks.
Open Field Observation: Rat was placed (one at a time) in an open arena, on a flat surface with a clea
n absorbent paper and observed for at least 2 minutes. Absorbent paper was replaced for each group.
During this observation period, rat was evaluated as it moves about freely/unperturbed and the fol
lowing observations were made and observations were recorded using score/ranks:
gait
posture
tremors
mobility score
arousal level
clonic or tonic movements
stereotypic behaviour
bizarre behaviour
urination
defecation
rearing
abnormal vocalizations
Functional Tests: Functional testing includes motor activity, sensory evaluation, landing hindlimbs footsplay and measurement of grip performance.
Motor Activity: The motor activity of rats was measured using an automated animal activity measuring system (Make: Columbus Instruments) equipped with a computer analyzer. Each rat was individually placed in the activity cages of the instrument. The rats were monitored for 30 minutes. During this motor activity measurement session, parameters viz., the stereotypic time (small movements) in seconds, the ambulatory time (large ambulatory movement) in seconds, horizontal counts, a mbulatory counts were monitored. The Opto-Varimex 4 motor activity measurement system provided the data at 1 minute interval and the data was analyzed in blocks of 10 minutes interval and the same was reported.
Sensory Reactivity Measurements: After the 2 minutes (approximately) observation period, while the rat was in the open field arena, the following tests were conducted. The rat was allowed to move freely in the open field box for these tests but positioned in the box by the observer in order to administer stimulus. During sensory reactivity measurements, rats were observed for following and the observations were recorded using scores/ranks.
approach response
touch response
click response
tail-pinch response
pupil response
aerial righting reflex
Landing Hindlimbs Footsplay: The landing hind limbs foot splay was performed by dropping the rat onto a horizontal surface of the table top from a short height and measuring the distance between the hind feet upon landing. The hind feet of the rat were gently pressed to an ink pad just prior to testing. The rat was suspended in a prone position and then dropped from a height of approximately 30 cm on to a SOP format, which contains the details such as Study no., Animal no, Group and Sex. A clean recording SOP format was used for each rat. A total of 3 readings were recorded for each rat and average of 3 footsplay values is presented in the report along with the individual footsplay values.
Grip Performance: Hindlimbs and forelimbs grip performance was tested using computerized dual grip strength meter (Model: Columbus Instruments). Three trials were conducted for each rat i.e., three trials each for forelimb and hind limbs. Averages of three trials for both forelimb and hindlimbs are calculated and presented in the report along with the individual grip strength values.
Physiological Observations: Body temperature (rectal temperature) was measured in degree Celsius (°C) using digital thermometer. At the end of the functional test, body weight of each rat was measured.
Body Weight: Individual body weights (g) was recorded prior to test item administration on Day 1 and weekly thereafter (± 2 day) for all groups of rats during treatment and recovery period. Fasting body weight was recorded prior to sacrifice for all animals.
Food Consumption: The food consumption was measured at weekly intervals (± 2 days) during treatment and recovery period. The cage wise average food consumption (g/rat/day) was calculated and presented in the report.
Oestrous Cycle Evaluation: Vaginal smear was examined in the female rats and the stage of oestrous cycle was recorded prior to necropsy.
Clinical Pathology Investigations
Blood Collection: At the end of the treatment and recovery periods, all rats were fasted overnight (water allowed) and approximately 4.0 mL of blood was collected under isoflurane anaesthesia, with a fine capillary tube, by retro-orbital sinus puncture: After analysis and data review by the analyst, the residual samples were disposed. Haematology, Coagulation, Clinical Chemistry, Hormone Analysis & Urinalysis Parameters were done as study plan. - Sacrifice and pathology:
- All rats from toxicity groups at the end of the scheduled period (Day 91 and 119) were subjected to detailed necropsy (examination of external surfaces of the body, all orifices; cranial, thoracic and abdominal cavities and their contents) and findings were recorded. Terminal fasting body weights were recorded for all animals immediately prior to terminal sacrifice and used in calculation of relative organ weights. All rats sacrificed at term were fasted overnight (water allowed), euthanized with isoflurane (as per the random numbers generated for the study), exsanguinated and subjected for gross examination.
At sacrifice, sperm motility was evaluated using the sperm samples collected from the right vas deferens, immediately after necropsy using Hamilton-Thorne TOX-IVOS sperm analyzer for all rats. For morphological evaluation of sperms, smears were made using semen samples collected from right vas deferens of all rats immediately after necropsy and fixed with acetone for evaluation by manual method. Initially, sperm morphology was assessed for control and high dose rats. The right epididymis was collected and frozen for enumeration of cauda epididymal sperm reserves. As the high dose group did not show any test item-related effect, the analysis was not extended to low, mid dose and recovery rats. Unused frozen samples were discarded at the time of final report preparation On completion of gross pathology examination, the tissues/organs noted in following table were collected from all rats. The below listed organs were weighed from all terminally sacrificed animals. The organ weight ratios (organ to body weight and brain weight) as percentage of fasting body weight
were determined and presented in the report. Paired organs were weighed together, and combined weights were presented.
Histopathological examination was carried out on the preserved organs of vehicle control (G1) and high dose group animals (G4). Examination of the testes also included a qualitative assessment of stages of spermatogenesis. In addition, all gross lesions from all the animals were examined microscopically. In the absence of test item-related changes, the tissues from low (G2), mid dose (G3) and recovery (G1R and G4R) groups were not evaluated.
The tissues were processed for routine paraffin embedding and 4-5-micron sections were stained with Haematoxylin and Eosin stain. In addition, testes were sectioned at 3-4 μm and stained with PAS reagent and haematoxylin to aid in qualitative assessment of spermatogenesis. Unused tissues were archived - Statistics:
- For comparative statistics, data were evaluated using the Levene Test for homogeneity of variances and the Shapiro-Wilks Test for normality of distributions. When data found to be homogeneous and of normal distribution, was analysed by analysis of variance (ANOVA), when data found to be no
nhomogeneous or of nonnormal data was subjected for transformation and ANOVA was done on transformed data. When ANOVA was significant, pairwise comparisons of treated groups to the control group was made using a parametric test, Dunnett, to identify statistical differences.
Data captured outside of Provantis™: The statistical analysis of the experimental data was carried out using licensed copies of SYSTAT Statistical package Ver.12.0. All quantitative variables neurological observations (neuromuscular observation/body temperature/body weights) and T3, T4, TSH was tested for normality (Shapiro-Wilk test) and homogeneity of variances (Levene’s test) within the group before performing a one-factor ANOVA modelling by treatment groups. Non-optimal (nonnormal or heteroschedastic) data was transformed, before ANOVA was performed. Comparison of means between treatment groups and control group was done using Dunnett’s test when the overall treatment, ‘F’ test was found significant.
For two groups, the comparisons of the mean between treatment and control group was done using‘t’ test.
Descriptive statistics (Mean, SD & Numbers) was presented by Treatment group and Day. All hypothesis testing were carried out at the 5% (2-sided) significance level. Significant differences are designated throughout the report as below:
*: Statistically significant difference from the control group at p < 0.05
Results and discussion
Results of examinations
- Clinical signs:
- effects observed, non-treatment-related
- Description (incidence and severity):
- There were no clinical signs and mortalities observed throughout the treatment and recovery period in either sex at all the doses tested. Light orange coloured faeces were observed at 111 and 333 mg/kg bwt/day doses and dark orange colour faeces were observed at 1000 mg/kg bwt/day doses in both sexes. This could be due to physical nature of the test item
- Mortality:
- no mortality observed
- Body weight and weight changes:
- effects observed, non-treatment-related
- Description (incidence and severity):
- Significantly lower body weight gain was observed from Days 22-29 at 333 mg/kg/day and during Days 64-71 at 1000 mg/kg/day in males and significantly higher body weight gain during Days 57-64 at 111 mg/kg/day in females. Significantly higher body weight gain was observed during Days 85-90, 90-97 and 90-118 at 1000 mg/kg/day recovery males and during Days 57-64 and 104-111 at 1000 mg/kg/day recovery females
- Food consumption and compound intake (if feeding study):
- effects observed, non-treatment-related
- Description (incidence and severity):
- The food consumption was significantly lower in males during Days 1-8 at 333 and during Days 15-22 at 111 and 333 mg/kg/day doses.
In females, the food consumption was significantly lower during Days 15-22 at all the doses in main toxicity groups and during Days 43-50 at 1000 mg/kg/day recovery females. Significantly higher food consumption was observed during Days 64-71 at all the doses in main toxicity groups and during Days 71-77 and 111-118 at 1000 mg/kg/day recovery females. - Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- not examined
- Ophthalmological findings:
- no effects observed
- Haematological findings:
- no effects observed
- Clinical biochemistry findings:
- no effects observed
- Endocrine findings:
- no effects observed
- Urinalysis findings:
- no effects observed
- Behaviour (functional findings):
- no effects observed
- Immunological findings:
- not examined
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Gross pathological findings:
- no effects observed
- Neuropathological findings:
- no effects observed
- Histopathological findings: non-neoplastic:
- no effects observed
- Histopathological findings: neoplastic:
- no effects observed
- Other effects:
- no effects observed
Effect levels
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- <= 1 000 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Basis for effect level:
- behaviour (functional findings)
- body weight and weight gain
- clinical biochemistry
- clinical signs
- food consumption and compound intake
- gross pathology
- haematology
- histopathology: non-neoplastic
- mortality
- ophthalmological examination
- organ weights and organ / body weight ratios
- urinalysis
Target system / organ toxicity
- Key result
- Critical effects observed:
- no
Any other information on results incl. tables
TABLE 2. Summary of Clinical Signs
Observation Type: All Types |
| Male |
|
| Female |
| ||
From Day 1 (Start Date) to 91 (End Date) | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day |
Normal Number of Animals Affected | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
First to Last seen | 1 - 91 | 1 - 1 | 1 - 1 | 1 - 1 | 1 - 91 | 1 - 1 | 1 - 1 | 1 - 1 |
Faecal colour Number of Animals Affected | 0 | 10 | 10 | 10 | 0 | 10 | 10 | 10 |
First to Last seen | - | 2 - 91 | 2 - 91 | 2 - 91 | - | 2 - 91 | 2 - 91 | 2 - 91 |
TABLE 2 contd. Summary of Clinical Signs
Observation Type: All Types | Male | Female | ||
From Day 1 (Start Date) to 119 (End Date) | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | G1R 0 mg/kg/day | G4R 1000 mg/kg/day |
Normal Number of Animals Affected | 5 | 5 |
5 | 5 |
First to Last seen | 1 - 119 | 1 - 119 | 1 - 119 | 1, 92 - 119 |
Faecal colour Number of Animals Affected |
0 | 5 | 0 | 5 |
First to Last seen | - | 2 - 91 | - | 2 - 91 |
TABLE 3. Summary of Ophthalmological Findings
Observation Type: All Types |
| Male |
|
| Female |
|
From Day -5 (Start Date) to 90 (End Date) | G1 0 mg/kg/day | G2 G3 111 333 mg/kg/day mg/kg/day | G4 1000 mg/kg/day | G1 0 mg/kg/day | G2 G3 111 333 mg/kg/day mg/kg/day | G4 1000 mg/kg/day |
Normal | 10 | 10 10 | 10 | 10 | 10 10 | 10 |
TABLE 3 contd. Summary of Ophthalmological Findings
From Day -5 (Start Date) to 119 (End Date) | G1R G4R 0 1000 mg/kg/day mg/kg/day | G1R G4R 0 1000 mg/kg/day mg/kg/day |
Normal | 5 5 | 5 5 |
TABLE 4. Summary of Functional Observation Battery - Males
| Group | G1 | G2 | G3 | G4 |
Parameters | Dose | 0 | 111 | 333 | 1000 |
| (mg/kg/day ) No. of rats | 10 | 10 | 10 | 10 |
Neurobehavioral Observations Home Cage Observations Posture Sitting normally, feet tucked in |
| 0 | 1 | 0 | 0 |
Sitting or Standing alert, watching |
| 10 | 8 | 8 | 6 |
Rears |
| 0 | 1 | 2 | 4 |
Abnormal Vocalizations Absent |
| 10 | 10 | 10 | 10 |
Tremors Absent |
| 10 | 10 | 10 | 10 |
Convulsions - Clonic Movement Absent |
| 10 | 10 | 10 | 10 |
Convulsions - Tonic Movement Absent |
| 10 |
10 |
10 |
10 |
Handling Observations Ease of Removing from Cage Easy (minimally avoids capture but is not aggressive) |
| 10 |
10 |
10 |
10 |
Handling Reactivity Moderately low (slight resistance) |
| 10 | 10 | 10 | 10 |
Palpebral Closure Eyelids wide open i.e., Normal |
| 10 | 10 | 10 | 10 |
Eye Examination Normal |
| 10 | 10 | 10 | 10 |
Piloerection Absent |
| 10 | 10 | 10 | 10 |
Lacrimation Absent |
| 10 | 10 | 10 | 10 |
Salivation None (no external salivation observed) |
| 10 | 10 | 10 | 10 |
| Group | G1 | G2 | G3 | G4 |
Parameters | Dose | 0 | 111 | 333 | 1000 |
| (mg/kg/day ) No. of rats | 10 | 10 | 10 | 10 |
Skin/Fur Examination Normal |
| 10 | 10 | 10 | 10 |
Perineum Wetness Absent |
| 10 | 10 | 10 | 10 |
Respiration Normal |
| 10 | 10 | 10 | 10 |
Muscle Tone Moderate (animal is neither very relaxed nor tense; Normal muscle tone) |
| 10 | 10 | 10 | 10 |
Extensor Thrust Response Moderate (clearly detectable; Normal) |
| 10 | 10 | 10 | 10 |
Open Field Observations Gait Normal gait |
| 10 | 10 | 10 | 10 |
Posture Normal posture |
| 10 | 10 | 10 | 10 |
Tremors Absent |
| 10 | 10 | 10 | 10 |
Mobility Score No impairment |
| 10 | 10 | 10 | 10 |
Arousal Level Alert (some exploratory movements; normal) |
| 10 | 10 | 10 | 10 |
Clonic Movement Absent |
| 10 | 10 | 10 | 10 |
Tonic Movement Absent |
| 10 | 10 | 10 | 10 |
TABLE 4 contd. Summary of Functional Observation Battery - Males
| Group | G1 | G2 | G3 | G4 |
Parameters | Dose | 0 | 111 | 333 | 1000 |
| (mg/kg/day ) No. of rats | 10 | 10 | 10 | 10 |
Stereotypic Behavior Absent |
| 10 | 10 | 10 | 10 |
Bizarre Behavior Absent |
| 10 | 10 | 10 | 10 |
Urination Absent |
| 8 | 8 | 7 | 7 |
Normal |
| 2 | 2 | 3 | 3 |
Defecation Absent |
| 7 | 8 | 7 | 8 |
Normal |
| 3 | 2 | 3 | 2 |
Abnormal Vocalization Absent |
| 10 | 10 | 10 | 10 |
Rearing |
| 79 | 75 | 87 | 78 |
Sensory Reactivity Measurements Approach Response Normal response (approaches towards the object) |
| 10 | 10 | 10 | 10 |
Touch Response Normal Response (spontaneously lifts tail when touched) |
| 10 | 10 | 10 | 10 |
Click Response Normal Response (clear movements of the head, neck and ears) |
| 10 | 10 | 10 | 10 |
Tail-Pinch Response Normal Response (Flinches, moves rapidly and/or vocalizes) |
| 10 | 10 | 10 | 10 |
Pupil Response Normal |
| 10 | 10 | 10 | 10 |
Aerial Righting Reflex Normal |
| 10 | 10 | 10 | 10 |
| Group | G1 | G2 | G3 | G4 |
Parameters | Dose | 0 | 111 | 333 | 1000 |
| (mg/kg/day ) No. of rats | 10 | 10 | 10 | 10 |
Physiological Observation Body Temperature (°C) | Mean | 37.55 | 37.66 | 37.86 | 37.58 |
| SD | 0.58 | 0.41 | 0.28 | 0.50 |
Body Weight (g) | Mean | 538.50 | 504.77 | 516.38 | 518.73 |
| SD | 49.90 | 57.70 | 42.90 | 38.18 |
Hindlimbs Footsplay (mm) Average | Mean | 80.70 | 80.30 | 84.40 | 64.17* |
| SD | 11.88 | 12.77 | 7.82 | 12.07 |
Grip Strength (gf) Forelimbs Grip Strength Average | Mean | 1056.60 | 1050.43 | 1051.30 | 1055.83 |
| SD | 21.83 | 18.48 | 11.88 | 19.60 |
Hindlimbs Grip Strength Average | Mean | 832.37 | 842.43 | 829.10 | 832.43 |
| SD | 19.75 | 24.51 | 20.25 | 20.33 |
*: Statistically significant difference from the control group at p < 0.05
TABLE 4 contd. Summary of Functional Observation Battery - Males
Group |
| Ambulatory (s) | Ambulatory (s) | Ambulatory (s) | Ambulatory (s) | Stereotypic (s) | Stereotypic (s) | Stereotypic (s) | Stereotypic |
Dose | (s) | ||||||||
(mg/kg/day) |
| 1 | 2 | 3 | Total | 1 | 2 | 3 | Total |
G1 | Mean SD N | 562.48 39.85 10 | 496.00 87.62 10 | 473.16 67.89 10 | 1531.64 173.34 10 | 27.81 32.61 10 | 57.76 47.67 10 | 52.81 26.99 10 | 138.38 |
0 | 76.38 | ||||||||
| 10 | ||||||||
|
|
|
|
|
|
|
|
|
|
G2 | Mean SD | 547.48 43.20 | 466.52 71.06 | 440.67 70.16 | 1454.67 163.88 | 30.53 21.69 | 76.73 42.33 | 92.52 55.78 | 199.78 |
111 | 99.14 | ||||||||
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
|
|
|
|
|
|
|
|
|
|
G3 | Mean SD | 577.55 20.35 | 527.13 57.62 | 451.99 98.02 | 1556.67 139.04 | 15.25 15.29 | 41.06 38.03 | 66.07 38.71 | 122.38 |
333 | 66.33 | ||||||||
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
|
|
|
|
|
|
|
|
|
|
G4 | Mean SD | 572.77 22.98 | 514.99 33.43 | 441.96 62.63 | 1529.72 70.77 | 15.01 12.63 | 42.36 23.84 | 78.80 35.14 | 136.17 |
1000 | 44.04 | ||||||||
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
Group |
| Resting (s) | Resting (s) | Resting (s) | Resting (s) | Distance (cm) | Distance (cm) | Distance (cm) | Distance |
Dose | (cm) | ||||||||
(mg/kg/day) |
| 1 | 2 | 3 | Total | 1 | 2 | 3 | Total |
G1 | Mean SD | 9.71 9.23 | 46.24 47.39 | 73.48 55.35 | 129.43 99.64 | 1564.47 298.45 | 856.78 283.03 | 737.11 189.83 | 3158.36 |
0 | 720.22 | ||||||||
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
|
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|
|
|
G2 | Mean SD | 21.99 25.19 | 56.75 43.75 | 66.79 36.01 | 145.53 98.51 | 1207.77* 351.84 | 746.01 235.08 | 628.11 211.42 | 2581.89 |
111 | 743.02 | ||||||||
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
|
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|
G3 | Mean SD | 7.20 6.73 | 31.81 21.33 | 81.94 71.96 | 120.95 86.23 | 1742.36 378.46 | 1009.42 288.97 | 691.11 253.88 | 3442.89 |
333 | 828.09 | ||||||||
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
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|
|
G4 | Mean SD | 12.22 10.82 | 42.65 20.82 | 79.12 43.05 | 133.99 48.11 | 1410.36 232.95 | 831.16 144.06 | 624.91 116.49 | 2866.43 |
1000 | 416.98 | ||||||||
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
*: Statistically significant difference from the control group at p < 0.05
| Group | G1R | G4R |
Parameters | Dose | 0 | 1000 |
| (mg/kg/day ) No. of rats | 5 | 5 |
Neurobehavioral Observations Home Cage Observations Posture Sitting or Standing alert, watching |
| 5 | 5 |
Abnormal Vocalizations Absent |
| 5 | 5 |
Tremors Absent |
| 5 | 5 |
Convulsions - Clonic Movement Absent |
| 5 | 5 |
Convulsions - Tonic Movement Absent |
| 5 |
5 |
Handling Observations Ease of Removing from Cage Easy (minimally avoids capture but is not aggressive) |
| 5 |
5 |
Handling Reactivity Moderately low (slight resistance) |
| 5 | 5 |
Palpebral Closure Eyelids wide open i.e., Normal |
| 5 | 5 |
Eye Examination Normal |
| 5 | 5 |
Piloerection Absent |
| 5 | 5 |
Lacrimation Absent |
| 5 | 5 |
Salivation None (no external salivation observed) |
| 5 | 5 |
Skin/Fur Examination Normal |
| 5 | 5 |
| Group | G1R | G4R |
Parameters | Dose | 0 | 1000 |
| (mg/kg/day ) No. of rats | 5 | 5 |
Perineum Wetness Absent |
| 5 | 5 |
Respiration Normal |
| 5 | 5 |
Muscle Tone Moderate (animal is neither very relaxed nor tense; Normal muscle tone) |
| 5 | 5 |
Extensor Thrust Response Moderate (clearly detectable; Normal) |
| 5 | 5 |
Open Field Observations Gait Normal gait |
| 5 | 5 |
Posture Normal posture |
| 5 | 5 |
Tremors Absent |
| 5 | 5 |
Mobility Score No impairment |
| 5 | 5 |
Arousal Level Alert (some exploratory movements; normal) |
| 5 | 5 |
Clonic Movement Absent |
| 5 | 5 |
Tonic Movement Absent |
| 5 | 5 |
Stereotypic Behavior Absent |
| 5 | 5 |
| Group | G1R | G4R |
Parameters | Dose | 0 | 1000 |
| (mg/kg/day ) No. of rats | 5 | 5 |
Bizarre Behavior Absent |
| 5 | 5 |
Urination Absent |
| 2 | 2 |
Normal |
| 3 | 3 |
Defecation Absent |
| 1 | 1 |
Normal |
| 4 | 4 |
Abnormal Vocalization Absent |
| 5 | 5 |
Rearing |
| 33 | 30 |
Sensory Reactivity Measurements Approach Response Normal response (approaches towards the object) |
| 5 | 5 |
Touch Response Normal Response (spontaneously lifts tail when touched) |
| 5 | 5 |
Click Response Normal Response (clear movements of the head, neck and ears) |
| 5 | 5 |
Tail-Pinch Response Normal Response (Flinches, moves rapidly and/or vocalizes) |
| 5 | 5 |
Pupil Response Normal |
| 5 | 5 |
Aerial Righting Reflex Normal |
| 5 | 5 |
| Group | G1R | G4R |
Parameters | Dose | 0 | 1000 |
| (mg/kg/day ) No. of rats | 5 | 5 |
Physiological Observation Body Temperature (°C) | Mean | 37.40 | 37.52 |
| SD | 0.61 | 0.55 |
Body Weight (g) | Mean | 555.67 | 590.44 |
| SD | 40.61 | 64.78 |
Hindlimbs Footsplay (mm) Average | Mean | 62.33 | 74.87 |
| SD | 9.46 | 22.12 |
Grip Strength (gf) Forelimbs Grip Strength Average | Mean | 1103.13 | 1068.00 |
| SD | 49.30 | 31.47 |
Hindlimbs Grip Strength Average | Mean | 776.40 | 836.80 |
| SD | 102.26 | 23.67 |
Group Dose |
| Ambulatory (s) | Ambulatory (s) | Ambulatory (s) | Ambulatory (s) | Stereotypic (s) | Stereotypic (s) | Stereotypic (s) | Stereotypic |
(s) | |||||||||
(mg/kg/day) |
| 1 | 2 | 3 | Total | 1 | 2 | 3 | Total |
G1R | Mean | 518.76 | 379.62 | 313.86 | 1212.24 | 55.62 | 113.76 | 137.26 | 306.64 |
0
| SD N | 33.48 5 | 61.99 5 | 102.78 5 | 160.87 5 | 34.84 5 | 35.57 5 | 65.65 5 | 113.65 |
5 | |||||||||
G4R 1000 |
Mean SD |
566.16 34.47 |
458.08 142.91 |
315.38 92.22 |
1339.62 198.39 |
15.98* 14.36 |
57.46 45.63 |
96.52 29.14 |
169.96 |
69.72 | |||||||||
| N | 5 | 5 | 5 | 5 | 5 | 5 | 5 | 5 |
Group |
| Resting (s) | Resting (s) | Resting (s) | Resting (s) | Distance (cm) | Distance (cm) | Distance (cm) | Distance |
Dose | (cm) | ||||||||
(mg/kg/day) |
| 1 | 2 | 3 | Total | 1 | 2 | 3 | Total |
G1R | Mean SD | 25.62 14.49 | 106.62 33.33 | 148.88 85.80 | 281.12 104.81 | 1177.02 281.74 | 534.03 107.09 | 392.82 187.51 | 2103.87 |
0 | 440.71 | ||||||||
| N | 5 | 5 | 5 | 5 | 5 | 5 | 5 | 5 |
|
Mean SD N |
17.86 20.34 5 |
84.46 103.72 5 |
188.10 96.49 5 |
290.42 161.83 5 |
1445.34 187.64 5 |
796.46 283.20 5 |
480.82 222.72 5 |
|
G4R | 2722.63 | ||||||||
1000 | 479.13 | ||||||||
| 5 |
*: Statistically significant difference from the control group at p < 0.05
TABLE 5. Summary of Functional Observation Battery - Females
| Group | G1 | G2 | G3 | G4 |
Parameters | Dose | 0 | 111 | 333 | 1000 |
| (mg/kg/day ) No. of rats | 10 | 10 | 10 | 10 |
Neurobehavioral Observations Home Cage Observations Posture Sitting normally, feet tucked in |
| 0 | 2 | 0 | 0 |
Sitting or Standing alert, watching |
| 6 | 7 | 7 | 7 |
Rears |
| 4 | 1 | 3 | 3 |
Abnormal Vocalizations Absent |
| 10 | 10 | 10 | 10 |
Tremors Absent |
| 10 | 10 | 10 | 10 |
Convulsions - Clonic Movement Absent |
| 10 | 10 | 10 | 10 |
Convulsions - Tonic Movement Absent |
| 10 |
10 |
10 |
10 |
Handling Observations Ease of Removing from Cage Easy (minimally avoids capture but is not aggressive) |
| 10 |
10 |
10 |
10 |
Handling Reactivity Moderately low (slight resistance) |
| 10 | 10 | 10 | 10 |
Palpebral Closure Eyelids wide open i.e., Normal |
| 10 | 10 | 10 | 10 |
Eye Examination Normal |
| 10 | 10 | 10 | 10 |
Piloerection Absent |
| 10 | 10 | 10 | 10 |
Lacrimation Absent |
| 10 | 10 | 10 | 10 |
Salivation None (no external salivation observed) |
| 10 | 10 | 10 | 10 |
| Group | G1 | G2 | G3 | G4 |
Parameters | Dose | 0 | 111 | 333 | 1000 |
| (mg/kg/day ) No. of rats | 10 | 10 | 10 | 10 |
Skin/Fur Examination Normal |
| 10 | 10 | 10 | 10 |
Perineum Wetness Absent |
| 10 | 10 | 10 | 10 |
Respiration Normal |
| 10 | 10 | 10 | 10 |
Muscle Tone Moderate (animal is neither very relaxed nor tense; Normal muscle tone) |
| 10 | 10 | 10 | 10 |
Extensor Thrust Response Moderate (clearly detectable; Normal) |
| 10 | 10 | 10 | 10 |
Open Field Observations Gait Normal gait |
| 10 | 10 | 10 | 10 |
Posture Normal posture |
| 10 | 10 | 10 | 10 |
Tremors Absent |
| 10 | 10 | 10 | 10 |
Mobility Score No impairment |
| 10 | 10 | 10 | 10 |
Arousal Level Alert (some exploratory movements; normal) |
| 10 | 10 | 10 | 10 |
Clonic Movement Absent |
| 10 | 10 | 10 | 10 |
Tonic Movement Absent |
| 10 | 10 | 10 | 10 |
TABLE 5 contd. Summary of Functional Observation Battery - Females
| Group | G1 | G2 | G3 | G4 |
Parameters | Dose | 0 | 111 | 333 | 1000 |
| (mg/kg/day ) No. of rats | 10 | 10 | 10 | 10 |
Stereotypic Behavior Absent |
| 10 | 10 | 10 | 10 |
Bizarre Behavior Absent |
| 10 | 10 | 10 | 10 |
Urination Absent |
| 8 | 6 | 8 | 10 |
Normal |
| 2 | 4 | 2 | 0 |
Defecation Absent |
| 7 | 7 | 9 | 7 |
Normal |
| 3 | 3 | 1 | 3 |
Abnormal Vocalization Absent |
| 10 | 10 | 10 | 10 |
Rearing |
| 81 | 101 | 99 | 88 |
Sensory Reactivity Measurements Approach Response Normal response (approaches towards the object) |
| 10 | 10 | 10 | 10 |
Touch Response Normal Response (spontaneously lifts tail when touched) |
| 10 | 10 | 10 | 10 |
Click Response Normal Response (clear movements of the head, neck and ears) |
| 10 | 10 | 10 | 10 |
Tail-Pinch Response Normal Response (Flinches, moves rapidly and/or vocalizes) |
| 10 | 10 | 10 | 10 |
Pupil Response Normal |
| 10 | 10 | 10 | 10 |
Aerial Righting Reflex Normal |
| 10 | 10 | 10 | 10 |
| Group | G1 | G2 | G3 | G4 |
Parameters | Dose | 0 | 111 | 333 | 1000 |
| (mg/kg/day ) No. of rats | 10 | 10 | 10 | 10 |
Physiological Observation Body Temperature (°C) | Mean | 37.61 | 37.76 | 37.68 | 37.79 |
| SD | 0.71 | 0.46 | 0.58 | 0.53 |
Body Weight (g) | Mean | 299.72 | 307.34 | 296.45 | 307.67 |
| SD | 22.55 | 25.46 | 19.18 | 27.51 |
Hindlimbs Footsplay (mm) Average | Mean | 63.87 | 67.67 | 70.53 | 74.60 |
| SD | 12.85 | 10.88 | 12.01 | 7.54 |
Grip Strength (gf) Forelimbs Grip Strength Average | Mean | 947.90 | 951.23 | 950.20 | 950.40 |
| SD | 21.48 | 18.06 | 17.12 | 21.27 |
Hindlimbs Grip Strength Average | Mean | 758.63 | 760.00 | 754.77 | 763.77 |
| SD | 16.38 | 15.35 | 16.53 | 23.18 |
TABLE 5 contd. Summary of Functional Observation Battery - Females
Group Dose (mg/kg/day) |
| Ambulatory (s) 1 | Ambulatory (s) 2 | Ambulatory (s) 3 | Ambulatory (s) Total | Stereotypic (s) 1 | Stereotypic (s) 2 | Stereotypic (s) 3 | Stereotypic |
(s) | |||||||||
Total | |||||||||
G1 0
| Mean SD N | 580.56 9.73 10 | 539.54 43.98 10 | 490.11 62.84 10 | 1610.21 103.97 10 | 10.25 5.81 10 | 33.25 31.16 10 | 57.97 35.43 10 | 101.47 |
58.76 | |||||||||
10 | |||||||||
|
|
|
|
|
|
|
|
|
|
G2 | Mean | 587.18 | 566.80 | 500.52 | 1654.50 | 7.96 | 22.78 | 54.54 | 85.28 |
111
| SD N | 9.40 10 | 30.44 10 | 65.62 10 | 80.05 10 | 7.92 10 | 25.53 10 | 41.34 10 | 51.89 |
10 | |||||||||
|
|
|
|
|
|
|
|
|
|
G3 333 | Mean SD | 585.38 8.05 | 549.03 41.21 | 467.99 140.35 | 1602.40 160.70 | 8.36 2.64 | 25.29 23.24 | 37.62 31.45 | 71.27 |
45.03 | |||||||||
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
|
|
|
|
|
|
|
|
|
|
G4 1000 | Mean SD | 588.20 7.70 | 570.03 17.37 | 518.42 61.46 | 1676.65 56.57 | 6.57 6.02 | 17.50 14.53 | 39.14 26.23 | 63.21 |
24.21 | |||||||||
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
Group |
| Resting (s) | Resting (s) | Resting (s) | Resting (s) | Distance (cm) | Distance (cm) | Distance (cm) | Distance |
Dose | (cm) | ||||||||
(mg/kg/day) |
| 1 | 2 | 3 | Total | 1 | 2 | 3 | Total |
G1 | Mean SD | 9.19 6.67 | 27.21 19.83 | 51.92 35.39 | 88.32 53.92 | 1838.31 271.63 | 1186.30 234.07 | 956.16 229.56 | 3980.76 |
0 | 653.60 | ||||||||
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
|
|
|
|
|
|
|
|
|
|
G2 | Mean SD | 4.86 2.64 | 10.42 8.01 | 44.93 37.05 | 60.21 37.41 | 1785.44 291.63 | 1358.14 412.32 | 1028.15 395.26 | 4171.73 |
111 | 976.26 | ||||||||
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
|
|
|
|
|
|
|
|
|
|
G3 | Mean SD | 6.26 7.10 | 25.68 24.27 | 94.39 143.33 | 126.33 158.22 | 1776.21 291.05 | 1313.50 537.21 | 966.97 225.28 | 4056.68 |
333 | 823.08 | ||||||||
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
|
|
|
|
|
|
|
|
|
|
G4 | Mean SD | 5.23 2.90 | 12.47 7.85 | 42.44 46.07 | 60.14 44.50 | 1913.65 275.02 | 1227.80 256.27 | 1095.84 305.09 | 4237.28 |
1000 | 744.67 | ||||||||
| N | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
| Group | G1R | G4R |
Parameters | Dose | 0 | 1000 |
| (mg/kg/day ) No. of rats | 5 | 5 |
Neurobehavioral Observations Home Cage Observations Posture Sitting or Standing alert, watching |
| 4 | 3 |
Rears |
| 1 | 2 |
Abnormal Vocalizations Absent |
| 5 | 5 |
Tremors Absent |
| 5 | 5 |
Convulsions - Clonic Movement Absent |
| 5 | 5 |
Convulsions - Tonic Movement Absent |
| 5 |
5 |
Handling Observations Ease of Removing from Cage Easy (minimally avoids capture but is not aggressive) |
| 5 |
5 |
Handling Reactivity Moderately low (slight resistance) |
| 5 | 5 |
Palpebral Closure Eyelids wide open i.e., Normal |
| 5 | 5 |
Eye Examination Normal |
| 5 | 5 |
Piloerection Absent |
| 5 | 5 |
Lacrimation Absent |
| 5 | 5 |
Salivation None (no external salivation observed) |
| 5 | 5 |
| Group | G1R | G4R |
Parameters | Dose | 0 | 1000 |
| (mg/kg/day ) No. of rats | 5 | 5 |
Skin/Fur Examination Normal |
| 5 | 5 |
Perineum Wetness Absent |
| 5 | 5 |
Respiration Normal |
| 5 | 5 |
Muscle Tone Moderate (animal is neither very relaxed nor tense; Normal muscle tone) |
| 5 | 5 |
Extensor Thrust Response Moderate (clearly detectable; Normal) |
| 5 | 5 |
Open Field Observations Gait Normal gait |
| 5 | 5 |
Posture Normal posture |
| 5 | 5 |
Tremors Absent |
| 5 | 5 |
Mobility Score No impairment |
| 5 | 5 |
Arousal Level Alert (some exploratory movements; normal) |
| 5 | 5 |
Clonic Movement Absent |
| 5 | 5 |
Tonic Movement Absent |
| 5 | 5 |
| Group | G1R | G4R |
Parameters | Dose | 0 | 1000 |
| (mg/kg/day ) No. of rats | 5 | 5 |
Stereotypic Behavior Absent |
| 5 | 5 |
Bizarre Behavior Absent |
| 5 | 5 |
Urination Absent |
| 3 | 4 |
Normal |
| 2 | 1 |
Defecation Absent |
| 2 | 2 |
Normal |
| 3 | 3 |
Abnormal Vocalization Absent |
| 5 | 5 |
Rearing |
| 70 | 65 |
Sensory Reactivity Measurements Approach Response Normal response (approaches towards the object) |
| 5 | 5 |
Touch Response Normal Response (spontaneously lifts tail when touched) |
| 5 | 5 |
Click Response Normal Response (clear movements of the head, neck and ears) |
| 5 | 5 |
Tail-Pinch Response Normal Response (Flinches, moves rapidly and/or vocalizes) |
| 5 | 5 |
Pupil Response Normal |
| 5 | 5 |
Aerial Righting Reflex Normal |
| 5 | 5 |
| Group | G1R | G4R |
Parameters | Dose | 0 | 1000 |
| (mg/kg/day ) No. of rats | 5 | 5 |
Physiological Observation Body Temperature (°C) | Mean | 37.62 | 37.54 |
| SD | 0.72 | 0.61 |
Body Weight (g) | Mean | 317.27 | 332.23 |
| SD | 21.16 | 27.29 |
Hindlimbs Footsplay (mm) Average | Mean | 59.67 | 61.13 |
| SD | 8.92 | 18.04 |
Grip Strength (gf) Forelimbs Grip Strength Average | Mean | 959.87 | 967.00 |
| SD | 21.90 | 9.45 |
Hindlimbs Grip Strength Average | Mean | 776.87 | 755.80 |
| SD | 8.37 | 29.40 |
TABLE 5 contd. Summary of Functional Observation Battery - Females
Group |
| Ambulatory | Ambulatory | Ambulatory | Ambulatory | Stereotypic | Stereotypic | Stereotypic | Stereotypic |
Dose (mg/kg/day) |
| (s) 1 | (s) 2 | (s) 3 | (s) Total | (s) 1 | (s) 2 | (s) 3 | (s) |
Total | |||||||||
G1R | Mean | 574.32 | 489.90 | 378.70 | 1442.92 | 11.64 | 56.02 | 94.34 | 162.00 |
0
| SD N | 7.81 5 | 48.21 5 | 83.12 5 | 118.25 5 | 2.82 5 | 41.81 5 | 43.51 5 | 72.46 |
5 | |||||||||
|
|
|
|
|
|
|
|
|
|
G4R 1000 | Mean SD | 588.42* 10.95 | 509.68 74.72 | 467.60 80.69 | 1565.70 145.94 | 7.54 7.26 | 37.02 38.02 | 68.00 49.62 | 112.56 |
72.33 | |||||||||
| N | 5 | 5 | 5 | 5 | 5 | 5 | 5 | 5 |
Group |
| Resting (s) | Resting (s) | Resting Resting (s) (s) | Distance (cm) | Distance (cm) | Distance (cm) | Distance |
Dose | (cm) | |||||||
(mg/kg/day) |
| 1 | 2 | Total 195.04 71.26 64.81 | 240.77 | 279.15 | 3 | Total |
G1R | Mean SD | 14.04 6.28 | 54.08 21.21 | 741.31 287.32 | 3622.98 | |||
0 | 687.48 | |||||||
| N | 5 | 5 | 5 5 | 5 | 5 | 5 | 5 |
|
|
|
|
|
|
|
|
|
G4R | Mean SD | 4.04* 4.11 | 53.30 41.51 | 64.40 121.74 46.19 88.53 | 2081.03 451.03 | 1322.80 499.57 | 1060.48 537.62 | 4464.31 |
1000 | 1404.21 | |||||||
| N | 5 | 5 | 5 5 | 5 | 5 | 5 | 5 |
*: Statistically significant difference from the control group at p < 0.05
TABLE 6. Summary of Body Weights (g) - Males
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Body Weight (g) | 1 [a] | Mean SD N | 218.08 14.07 10 | 220.54 14.92 10 | 218.80 12.99 10 | 217.74 16.30 10 |
8 [a] | Mean SD N | 277.13 17.26 10 | 275.45 20.10 10 | 276.74 14.06 10 | 278.07 19.28 10 | |
15 [a] | Mean SD N | 324.70 23.99 10 | 313.65 28.16 10 | 320.05 17.34 10 | 321.51 20.76 10 | |
22 [a] | Mean SD N | 357.34 32.77 10 | 345.26 33.93 10 | 351.54 20.79 10 | 352.58 25.08 10 | |
29 [a] | Mean SD N | 386.64 37.82 10 | 370.53 37.61 10 | 374.07 23.87 10 | 380.39 24.28 10 | |
36 [a] | Mean SD N | 413.80 41.11 10 | 394.37 42.01 10 | 397.91 27.13 10 | 398.95 26.52 10 | |
43 [a] | Mean SD N | 436.51 40.21 10 | 417.85 45.34 10 | 423.64 28.30 10 | 425.45 28.89 10 | |
50 [a] | Mean SD N | 455.15 41.42 10 | 437.33 47.17 10 | 443.68 30.34 10 | 449.49 31.90 10 | |
57 [a] | Mean SD N | 474.55 43.08 10 | 452.63 50.71 10 | 462.34 32.89 10 | 462.07 32.70 10 | |
64 [a] | Mean SD N | 493.42 44.76 10 | 465.77 51.27 10 | 475.57 35.96 10 | 482.16 32.91 10 | |
71 [a] | Mean SD N | 517.22 48.83 10 | 484.75 53.08 10 | 492.49 40.32 10 | 497.64 35.47 10 | |
77 [a] | Mean SD N | 523.90 48.88 10 | 493.83 54.56 10 | 503.73 41.62 10 | 505.86 33.72 10 | |
[a] Anova & Dunnett
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Body Weight (g) | 85 [a] | Mean SD N | 538.82 50.88 10 | 507.26 56.70 10 | 514.63 40.20 10 | 521.97 39.20 10 |
90 [a] | Mean SD N | 538.16 48.63 10 | 506.79 54.33 10 | 519.48 39.13 10 | 523.66 39.81 10 | |
[a] Anova & Dunnett
TABLE 6 contd. Summary of Body Weights (g) - Males
Sex: Male Da | y(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | |
Body Weight (g) | 1 [p] | Mean SD N | 221.02 19.58 5 | 218.50 18.56 5 |
8 [p] | Mean SD N | 272.49 22.71 5 | 278.52 11.47 5 | |
15 [p] | Mean SD N | 317.80 26.64 5 | 327.93 12.56 5 | |
22 [p] | Mean SD N | 350.26 27.96 5 | 363.61 16.86 5 | |
29 [p] | Mean SD N | 377.76 24.61 5 | 389.58 20.27 5 | |
36 [p] | Mean SD N | 401.23 36.12 5 | 416.43 22.48 5 | |
43 [p] | Mean SD N | 423.69 35.36 5 | 441.87 34.00 5 | |
50 [p] | Mean SD N | 443.31 35.95 5 | 464.16 38.13 5 | |
57 [p] | Mean SD N | 460.31 37.92 5 | 477.88 40.87 5 | |
64 [p] | Mean SD N | 478.10 39.77 5 | 500.82 46.14 5 | |
[p] T-Test: [G1RvsG4R]
Sex: Male Da | y(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | |
Body Weight (g) | 71 [p] | Mean SD N | 495.97 44.86 5 | 513.65 46.56 5 |
77 [p] | Mean SD N | 506.80 40.67 5 | 523.71 47.60 5 | |
85 [p] | Mean SD N | 521.03 36.89 5 | 533.97 55.30 5 | |
90 [p] | Mean SD N | 519.20 36.56 5 | 540.16 54.70 5 | |
97 [p] | Mean SD N | 526.24 30.84 5 | 561.16 51.99 5 | |
104 [p] | Mean SD N | 537.57 38.08 5 | 582.51 54.74 5 | |
111 [p] | Mean SD N | 540.87 41.76 5 | 585.11 59.04 5 | |
118 [p] | Mean SD N | 551.30 41.60 5 | 594.92 62.02 5 | |
[p] T-Test: [G1RvsG4R]
TABLE 7. Summary of Body Weights (g) - Females
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Body Weight (g) | 1 [a] | Mean SD N | 172.08 10.16 10 | 172.02 7.86 10 | 172.96 9.79 10 | 173.15 9.66 10 |
8 [a] | Mean SD N | 196.82 10.70 10 | 197.77 13.96 10 | 198.57 9.60 10 | 198.02 11.43 10 | |
15 [a] | Mean SD N | 212.31 14.04 10 | 216.29 13.49 10 | 212.78 14.71 10 | 213.99 15.68 10 | |
22 [a] | Mean SD N | 226.06 18.40 10 | 228.50 16.36 10 | 231.73 17.81 10 | 230.27 12.92 10 | |
29 [a] | Mean SD N | 243.52 18.19 10 | 242.32 20.46 10 | 239.73 14.78 10 | 246.53 16.41 10 | |
36 [a] | Mean SD N | 254.04 17.59 10 | 255.20 22.73 10 | 251.97 17.06 10 | 260.70 18.95 10 | |
43 [a] | Mean SD N | 257.81 18.07 10 | 261.73 24.36 10 | 260.23 16.31 10 | 265.95 22.02 10 | |
50 [a] | Mean SD N | 265.97 23.38 10 | 268.53 21.76 10 | 269.04 17.85 10 | 273.33 19.03 10 | |
57 [a] | Mean SD N | 279.85 23.84 10 | 275.92 22.77 10 | 276.82 20.23 10 | 286.57 21.62 10 | |
64 [a] | Mean SD N | 284.67 25.48 10 | 289.40 22.78 10 | 282.16 17.18 10 | 290.01 23.48 10 | |
71 [a] | Mean SD N | 286.41 22.56 10 | 295.32 24.30 10 | 289.17 17.53 10 | 300.62 28.02 10 | |
77 [a] | Mean SD N | 292.72 24.07 10 | 296.91 25.79 10 | 293.26 20.10 10 | 304.96 25.68 10 | |
[a] Anova & Dunnett
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Body Weight (g) | 85 [a] | Mean SD N | 298.17 23.87 10 | 303.26 24.33 10 | 293.32 15.93 10 | 308.37 26.22 10 |
90 [a] | Mean SD N | 296.83 24.63 10 | 304.10 26.35 10 | 296.94 17.12 10 | 309.35 29.99 10 | |
[a] Anova & Dunnett
TABLE 7 contd. Summary of Body Weights (g) - Females
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Body Weight (g) | 1 [p] | Mean SD N | 174.99 9.03 5 | 172.75 9.34 5 |
8 [p] | Mean SD N | 200.39 8.61 5 | 198.83 16.22 5 | |
15 [p] | Mean SD N | 221.36 9.33 5 | 219.11 16.04 5 | |
22 [p] | Mean SD N | 236.17 15.63 5 | 236.14 20.70 5 | |
29 [p] | Mean SD N | 245.95 11.50 5 | 254.24 19.94 5 | |
36 [p] | Mean SD N | 261.74 14.01 5 | 265.01 18.92 5 | |
43 [p] | Mean SD N | 265.04 15.70 5 | 271.90 21.29 5 | |
50 [p] | Mean SD N | 273.64 14.74 5 | 281.20 23.32 5 | |
57 [p] | Mean SD N | 287.58 15.43 5 | 289.35 25.88 5 | |
64 [p] | Mean SD N | 289.42 15.55 5 | 297.77 27.16 5 | |
[p] T-Test: [G1RvsG4R]
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Body Weight (g) | 71 [p] | Mean SD N | 298.17 19.97 5 | 303.90 26.63 5 |
77 [p] | Mean SD N | 300.72 19.80 5 | 311.95 24.64 5 | |
85 [p] | Mean SD N | 304.10 18.82 5 | 318.13 26.27 5 | |
90 [p] | Mean SD N | 302.68 20.51 5 | 317.65 27.65 5 | |
97 [p1] | Mean SD N | 314.60 19.98 5 | 325.45 29.24 5 | |
104 [p1] | Mean SD N | 312.44 17.22 5 | 327.57 30.50 5 | |
111 [p1] | Mean SD N | 309.30 17.85 5 | 328.40 28.31 5 | |
118 [p1] | Mean SD N | 311.68 20.69 5 | 332.15 24.24 5 | |
[p] - T-Test: [G1RvsG4R]
[p1] - Wilcoxon(Rank): [G1RvsG4R]
TABLE 8. Summary of Body Weight Gains (g) - Males
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Absolute Weight Gain (g) | 1 → 8 [a] | Mean SD N | 59.05 7.08 10 | 54.91 6.94 10 | 57.94 3.67 10 | 60.34 5.84 10 |
8 → 15 [a] | Mean SD N | 47.57 9.73 10 | 38.20 8.82 10 | 43.31 6.14 10 | 43.43 5.25 10 | |
15 → 22 [a] | Mean SD N | 32.65 9.73 10 | 31.61 6.80 10 | 31.49 5.54 10 | 31.07 6.58 10 | |
22 → 29 [a1] | Mean SD N | 29.30 6.29 10 | 25.27 6.44 10 | 22.53 * 5.02 10 | 27.81 2.46 10 | |
29 → 36 [a] | Mean SD N | 27.15 5.27 10 | 23.85 8.67 10 | 23.84 5.39 10 | 18.55 8.70 10 | |
36 → 43 [a] | Mean SD N | 22.71 5.22 10 | 23.48 5.70 10 | 25.73 2.89 10 | 26.50 5.21 10 | |
43 → 50 [a] | Mean SD N | 18.64 5.74 10 | 19.48 5.41 10 | 20.04 4.35 10 | 24.04 7.33 10 | |
50 → 57 [a1] | Mean SD N | 19.40 7.06 10 | 15.30 5.91 10 | 18.66 4.96 10 | 12.58 12.02 10 | |
57 → 64 [a] | Mean SD N | 18.87 8.78 10 | 13.14 5.26 10 | 13.23 8.62 10 | 20.09 9.47 10 | |
64 → 71 [a] | Mean SD N | 23.80 7.36 10 | 18.98 5.04 10 | 16.92 7.28 10 | 15.48 * 5.94 10 | |
71 → 77 [a1] | Mean SD N | 6.68 7.35 10 | 9.08 5.22 10 | 11.24 4.73 10 | 8.22 5.41 10 | |
77 → 85 [a] | Mean SD N | 14.92 6.70 10 | 13.43 7.60 10 | 10.90 3.96 10 | 16.11 7.68 10 | |
[a] - Anova & Dunnett: * = p < 0.05
[a1] - Anova & Dunnett(Rank): * = p < 0.05
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Absolute Weight Gain (g) | 85 → 90 [a] | Mean SD N | -0.66 7.56 10 | -0.47 5.86 10 | 4.85 6.31 10 | 1.70 6.48 10 |
1 → 90 [a] | Mean SD N | 320.08 40.81 10 | 286.25 42.26 10 | 300.68 33.90 10 | 305.93 29.17 10 | |
Total % Weight Gain | 1 → 90 [a] | Mean SD N | 146.83 16.75 10 | 129.47 13.37 10 | 137.67 15.89 10 | 140.80 12.88 10 |
[a] - Anova & Dunnett; [a1] - Anova & Dunnett(Log); [a2] - Anova & Dunnett(Rank) [a] - Anova & Dunnett
TABLE 8 contd. Summary of Body Weight Gains (g) - Males
Sex: Male Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Absolute Weight Gain (g) | 1 → 8 [p] | Mean SD N | 51.47 9.03 5 | 60.02 12.71 5 |
8 → 15 [p] | Mean SD N | 45.31 6.85 5 | 49.41 12.40 5 | |
15 → 22 [p] | Mean SD N | 32.46 5.55 5 | 35.68 10.11 5 | |
22 → 29 [p] | Mean SD N | 27.50 6.65 5 | 25.97 5.59 5 | |
29 → 36 [p] | Mean SD N | 23.47 12.55 5 | 26.85 3.65 5 | |
36 → 43 [p1] | Mean SD N | 22.46 3.12 5 | 25.44 12.30 5 | |
43 → 50 [p] | Mean SD N | 19.62 2.73 5 | 22.29 5.20 5 | |
50 → 57 [p] | Mean SD N | 17.01 5.83 5 | 13.72 7.19 5 | |
57 → 64 [p1] | Mean SD N | 17.78 3.21 5 | 22.94 11.89 5 | |
64 → 71 [p] | Mean SD N | 17.87 14.16 5 | 12.83 3.26 5 | |
[p] - T-Test: [G1RvsG4R]
[p1] - Wilcoxon(Rank): [G1RvsG4R]
Sex: Male Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Absolute Weight Gain (g) | 71 → 77 [p] | Mean SD N | 10.83 7.07 5 | 10.06 2.83 5 |
77 → 85 [p] | Mean SD N | 14.23 5.51 5 | 10.26 10.41 5 | |
85 → 90 [p] | Mean SD N | -1.83 2.04 5 | 6.19 * 3.06 5 | |
1 → 90 [p] | Mean SD N | 298.18 32.10 5 | 321.66 69.20 5 | |
90 → 97 [p] | Mean SD N | 7.04 8.00 5 | 21.00 * 10.02 5 | |
97 → 104 [p1] | Mean SD N | 11.33 8.64 5 | 21.35 19.80 5 | |
104 → 111 [p] | Mean SD N | 3.30 4.84 5 | 2.60 6.40 5 | |
111 → 118 [p] | Mean SD N | 10.43 10.50 5 | 9.82 7.84 5 | |
90 → 118 [p2] | Mean SD N | 32.10 8.14 5 | 54.76 * 21.68 5 | |
Total % Weight Gain | 1 → 90 [p] | Mean SD N | 135.75 18.68 5 | 149.73 41.14 5 |
[p] - T-Test: [G1RvsG4R]
{Absolute Wei}{85-90} G1RvsG4R: * = p < 0.05
{Absolute Wei}{90-97} G1RvsG4R: * = p < 0.05
[p1] - Wilcoxon(Rank): [G1RvsG4R]
[p2] - T-Test(Log): [G1RvsG4R]
{90-118} G1RvsG4R: * = p < 0.05
Sex: Male Da | y(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | |
Total % Weight Gain | 90 → 118 [p] | Mean SD N | 14.63 3.94 5 | 25.18 9.93 5 |
[p] - T-Test: [G1RvsG4R]
TABLE 9. Summary of Body Weight Gains (g) - Females
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Absolute Weight Gain (g) | 1 → 8 [a] | Mean SD N | 24.74 4.99 10 | 25.75 6.95 10 | 25.61 4.36 10 | 24.87 8.14 10 |
8 → 15 [a] | Mean SD N | 15.48 8.12 10 | 18.52 3.54 10 | 14.21 9.95 10 | 15.96 8.72 10 | |
15 → 22 [a] | Mean SD N | 13.75 11.23 10 | 12.21 8.40 10 | 18.96 10.08 10 | 16.28 11.95 10 | |
22 → 29 [a1] | Mean SD N | 17.46 2.86 10 | 13.82 10.86 10 | 7.99 9.54 10 | 16.27 7.86 10 | |
29 → 36 [a] | Mean SD N | 10.53 6.45 10 | 12.87 6.53 10 | 12.25 9.49 10 | 14.16 5.52 10 | |
36 → 43 [a1] | Mean SD N | 3.77 5.11 10 | 6.54 4.76 10 | 8.26 4.67 10 | 5.25 11.77 10 | |
43 → 50 [a] | Mean SD N | 8.16 10.55 10 | 6.79 6.26 10 | 8.81 7.74 10 | 7.38 10.27 10 | |
50 → 57 [a] | Mean SD N | 13.88 6.43 10 | 7.39 10.57 10 | 7.78 5.24 10 | 13.24 7.88 10 | |
57 → 64 [a1] | Mean SD N | 4.82 6.57 10 | 13.48 * 6.24 10 | 5.34 6.50 10 | 3.44 7.11 10 | |
64 → 71 [a1] | Mean SD N | 1.74 4.66 10 | 5.93 3.34 10 | 7.01 4.83 10 | 10.62 9.49 10 | |
71 → 77 [a] | Mean SD N | 6.31 6.77 10 | 1.58 4.05 10 | 4.09 7.81 10 | 4.34 6.72 10 | |
77 → 85 [a1] | Mean SD N | 5.45 4.22 10 | 6.36 4.80 10 | 0.06 8.05 10 | 3.41 2.94 10 | |
[a] - Anova & Dunnett
[a1] - Anova & Dunnett(Rank): * = p < 0.05
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Absolute Weight Gain (g) | 85 → 90 [a] | Mean SD N | -1.34 4.53 10 | 0.84 5.85 10 | 3.61 4.78 10 | 0.98 6.41 10 |
1 → 90 [a] | Mean SD N | 124.75 18.50 10 | 132.09 19.48 10 | 123.98 10.78 10 | 136.19 24.37 10 | |
Total % Weight Gain | 1 → 90 [a] | Mean SD N | 72.47 9.56 10 | 76.54 9.01 10 | 71.78 5.91 10 | 78.55 12.65 10 |
[a] - Anova & Dunnett
TABLE 9 contd. Summary of Body Weight Gains (g) - Females
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Absolute Weight Gain (g) | 85 → 90 [a] | Mean SD N | -1.34 4.53 10 | 0.84 5.85 10 | 3.61 4.78 10 | 0.98 6.41 10 |
1 → 90 [a] | Mean SD N | 124.75 18.50 10 | 132.09 19.48 10 | 123.98 10.78 10 | 136.19 24.37 10 | |
Total % Weight Gain | 1 → 90 [a] | Mean SD N | 72.47 9.56 10 | 76.54 9.01 10 | 71.78 5.91 10 | 78.55 12.65 10 |
[a] - Anova & Dunnett
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Absolute Weight Gain (g) | 1 → 8 [p] | Mean SD N | 25.39 4.18 5 | 26.08 8.15 5 |
8 → 15 [p] | Mean SD N | 20.97 8.07 5 | 20.28 5.18 5 | |
15 → 22 [p] | Mean SD N | 14.81 8.55 5 | 17.03 9.95 5 | |
22 → 29 [p] | Mean SD N | 9.78 12.34 5 | 18.11 21.89 5 | |
29 → 36 [p] | Mean SD N | 15.79 12.74 5 | 10.76 13.82 5 | |
36 → 43 [p] | Mean SD N | 3.31 6.77 5 | 6.89 9.41 5 | |
43 → 50 [p] | Mean SD N | 8.59 3.52 5 | 9.30 5.79 5 | |
50 → 57 [p] | Mean SD N | 13.95 8.67 5 | 8.15 5.36 5 | |
57 → 64 [p] | Mean SD N | 1.84 4.81 5 | 8.41 * 3.01 5 | |
64 → 71 [p] | Mean SD N | 8.75 6.43 5 | 6.13 8.26 5 | |
[p] - T-Test: [G1RvsG4R]
{57-64} G1RvsG4R: * = p < 0.05
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Absolute Weight Gain (g) | 71 → 77 [p] | Mean SD N | 2.55 7.36 5 | 8.05 5.08 5 |
77 → 85 [p] | Mean SD N | 3.39 2.10 5 | 6.18 3.01 5 | |
85 → 90 [p] | Mean SD N | -1.42 5.80 5 | -0.48 3.81 5 | |
1 → 90 [p] | Mean SD N | 127.69 15.27 5 | 144.90 24.64 5 | |
90 → 97 [p] | Mean SD N | 11.92 6.85 5 | 7.79 6.56 5 | |
97 → 104 [p] | Mean SD N | -2.16 4.40 5 | 2.12 7.75 5 | |
104 → 111 [p1] | Mean SD N | -3.14 1.63 5 | 0.84 * 4.36 5 | |
111 → 118 [p] | Mean SD N | 2.38 5.14 5 | 3.75 9.38 5 | |
90 → 118 [p] | Mean SD N | 9.00 9.61 5 | 14.50 5.16 5 | |
Total % Weight Gain | 1 → 90 [p] | Mean SD N | 72.99 8.00 5 | 83.97 13.56 5 |
[p] - T-Test: [G1RvsG4R]
[p1] - Wilcoxon(Rank): [G1RvsG4R]
{104-111} G1RvsG4R: * = p < 0.05
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Total % Weight Gain | 90 → 118 [p] | Mean SD N | 5.06 5.48 5 | 8.43 3.10 5 |
[p] - T-Test: [G1RvsG4R]
TABLE 10. Summary of Food Consumption (g/rat/day) - Males
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Food Consumption (g/animal/day) | 1 → 8 [a] | Mean SD N | 30.07 0.59 10 | 29.58 1.07 10 | 28.98 * 0.81 10 | 29.94 0.65 10 |
8 → 15 [a] | Mean SD N | 29.67 1.96 10 | 28.48 1.69 10 | 29.21 1.48 10 | 30.29 0.74 10 | |
15 → 22 [a] | Mean SD N | 29.70 1.14 10 | 28.14 * 1.82 10 | 28.07 * 0.82 10 | 29.19 1.26 10 | |
22 → 29 [a] | Mean SD N | 29.39 1.63 10 | 29.21 1.38 10 | 29.48 1.32 10 | 30.23 0.60 10 | |
29 → 36 [a] | Mean SD N | 30.18 1.30 10 | 29.28 1.68 10 | 29.01 1.13 10 | 30.67 1.19 10 | |
36 → 43 [a] | Mean SD N | 30.19 1.55 10 | 29.25 0.95 10 | 29.72 0.76 10 | 30.18 1.24 10 | |
43 → 50 [a1] | Mean SD N | 30.19 1.30 10 | 28.68 1.57 10 | 29.51 1.05 10 | 30.66 1.74 10 | |
50 → 57 [a] | Mean SD N | 31.86 1.06 10 | 31.18 2.10 10 | 31.64 1.65 10 | 31.68 1.44 10 | |
57 → 64 [a1] | Mean SD N | 32.16 1.41 10 | 31.08 2.28 10 | 31.20 1.26 10 | 32.31 1.94 10 | |
64 → 71 [a] | Mean SD N | 33.18 1.36 10 | 31.70 2.15 10 | 31.49 1.32 10 | 31.55 0.78 10 | |
71 → 77 [a] | Mean SD N | 32.00 1.53 10 | 31.02 1.31 10 | 30.81 1.23 10 | 31.38 1.46 10 | |
77 → 85 [a] | Mean SD N | 32.92 1.63 10 | 33.54 1.01 10 | 32.59 0.90 10 | 33.16 0.49 10 | |
[a] - Anova & Dunnett(Rank): * = p < 0.05
[a1] Anova & Dunnett
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Food Consumption (g/animal/day) | 85 → 90 [a] | Mean SD N | 31.12 1.60 10 | 31.04 1.25 10 | 31.75 1.95 10 | 30.82 2.09 10 |
[a] - Anova & Dunnett(Rank)
TABLE 10 contd. Summary of Food Consumption (g/rat/day) - Males
Sex: Male Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Food Consumption (g/animal/day) | 1 → 8 [p] | Mean SD N | 29.24 0.42 5 | 29.23 0.93 5 |
8 → 15 [p] | Mean SD N | 30.09 0.70 5 | 30.50 1.32 5 | |
15 → 22 [p] | Mean SD N | 28.86 0.89 5 | 29.19 0.99 5 | |
22 → 29 [p] | Mean SD N | 28.55 0.95 5 | 28.62 1.39 5 | |
29 → 36 [p] | Mean SD N | 29.13 0.65 5 | 29.58 1.19 5 | |
36 → 43 [p1] | Mean SD N | 29.03 0.24 5 | 30.05 1.21 5 | |
43 → 50 [p1] | Mean SD N | 29.36 0.73 5 | 28.72 0.78 5 | |
50 → 57 [p1] | Mean SD N | 31.67 0.27 5 | 30.78 0.93 5 | |
57 → 64 [p] | Mean SD N | 30.44 1.12 5 | 30.75 1.55 5 | |
64 → 71 [p1] | Mean SD N | 31.22 2.22 5 | 30.61 0.44 5 | |
[p] - T-Test: [G1RvsG4R]
[p1] Wilcoxon(Rank): [G1RvsG4R]
Sex: Male Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Food Consumption (g/animal/day) | 71 → 77 [p] | Mean SD N | 31.02 1.19 5 | 30.23 1.12 5 |
77 → 85 [p1] | Mean SD N | 32.82 0.79 5 | 32.50 0.32 5 | |
85 → 90 [p1] | Mean SD N | 30.79 1.00 5 | 31.29 1.48 5 | |
90 → 97 [p] | Mean SD N | 29.35 0.92 5 | 30.10 0.89 5 | |
97 → 104 [p1] | Mean SD N | 31.51 1.19 5 | 31.50 2.21 5 | |
104 → 111 [p1] | Mean SD N | 30.74 2.04 5 | 31.68 2.20 5 | |
111 → 118 [p1] | Mean SD N | 30.91 1.63 5 | 31.77 1.79 5 | |
[p] - T-Test: [G1RvsG4R]
[p1] Wilcoxon(Rank): [G1RvsG4R]
TABLE 11. Summary of Food Consumption (g/rat/day) - Females
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Food Consumption (g/animal/day) | 1 → 8 [a] | Mean SD N | 19.86 0.73 10 | 19.86 0.82 10 | 20.55 0.95 10 | 20.49 0.84 10 |
8 → 15 [a] | Mean SD N | 20.07 0.59 10 | 19.96 1.35 10 | 20.69 0.88 10 | 20.59 0.90 10 | |
15 → 22 [a] | Mean SD N | 20.74 0.15 10 | 19.93 * 0.85 10 | 20.27 * 0.41 10 | 19.99 * 0.35 10 | |
22 → 29 [a1] | Mean SD N | 20.81 0.56 10 | 20.65 0.83 10 | 20.60 0.49 10 | 21.01 0.67 10 | |
29 → 36 [a] | Mean SD N | 21.19 0.40 10 | 20.58 0.91 10 | 21.43 0.48 10 | 20.93 0.39 10 | |
36 → 43 [a] | Mean SD N | 21.06 0.49 10 | 20.87 0.88 10 | 20.40 1.64 10 | 20.14 0.96 10 | |
43 → 50 [a] | Mean SD N | 21.30 0.23 10 | 20.82 1.46 10 | 20.64 0.99 10 | 20.91 0.88 10 | |
50 → 57 [a] | Mean SD N | 22.09 0.33 10 | 21.81 1.12 10 | 22.39 0.21 10 | 22.55 0.29 10 | |
57 → 64 [a1] | Mean SD N | 22.57 0.96 10 | 22.67 0.96 10 | 22.63 0.43 10 | 23.35 0.81 10 | |
64 → 71 [a] | Mean SD N | 21.12 0.39 10 | 21.79 * 1.41 10 | 22.17 * 0.80 10 | 22.60 * 0.69 10 | |
71 → 77 [a] | Mean SD N | 22.27 0.55 10 | 21.76 1.24 10 | 21.43 0.98 10 | 22.11 0.69 10 | |
77 → 85 [a] | Mean SD N | 23.46 0.10 10 | 23.13 0.82 10 | 23.62 0.66 10 | 23.48 0.76 10 | |
[a] - Anova & Dunnett(Rank): * = p < 0.05
[a1] Anova & Dunnett
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Food Consumption (g/animal/day) | 85 → 90 [a] | Mean SD N | 22.91 0.40 10 | 22.15 1.30 10 | 22.16 1.13 10 | 22.81 0.95 10 |
[a] - Anova & Dunnett(Rank)
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Food Consumption (g/animal/day) | 1 → 8 [p] | Mean SD N | 20.12 0.99 5 | 19.66 0.22 5 |
8 → 15 [p] | Mean SD N | 20.88 0.29 5 | 20.25 1.34 5 | |
15 → 22 [p1] | Mean SD N | 20.00 0.63 5 | 19.65 0.78 5 | |
22 → 29 [p] | Mean SD N | 21.41 0.78 5 | 20.41 1.01 5 | |
29 → 36 [p] | Mean SD N | 21.63 0.05 5 | 21.16 0.45 5 | |
36 → 43 [p] | Mean SD N | 20.62 0.60 5 | 20.70 0.46 5 | |
43 → 50 [p1] | Mean SD N | 21.87 0.63 5 | 20.71 * 0.93 5 | |
50 → 57 [p1] | Mean SD N | 22.21 0.43 5 | 22.17 0.53 5 | |
57 → 64 [p] | Mean SD N | 22.99 1.17 5 | 22.95 0.48 5 | |
64 → 71 [p] | Mean SD N | 21.95 0.73 5 | 21.97 1.33 5 | |
[p] - Wilcoxon(Rank): [G1RvsG4R]
[p1] - T-Test: [G1RvsG4R]
{43-50} G1RvsG4R: * = p < 0.05
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Food Consumption (g/animal/day) | 71 → 77 [p] | Mean SD N | 21.30 0.90 5 | 22.29 * 0.37 5 |
77 → 85 [p] | Mean SD N | 23.11 0.59 5 | 23.22 1.17 5 | |
85 → 90 [p1] | Mean SD N | 22.74 1.01 5 | 21.75 1.50 5 | |
90 → 97 [p1] | Mean SD N | 22.64 0.39 5 | 22.30 0.64 5 | |
97 → 104 [p] | Mean SD N | 22.12 0.51 5 | 21.95 0.29 5 | |
104 → 111 [p] | Mean SD N | 22.75 0.75 5 | 22.88 0.15 5 | |
111 → 118 [p] | Mean SD N | 21.15 1.18 5 | 22.65 * 0.14 5 | |
[p] - Wilcoxon(Rank): [G1RvsG4R]
{71-77} G1RvsG4R: * = p < 0.05
{111-118} G1RvsG4R: * = p < 0.05
TABLE 12. Summary of Oestrous Cycle
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Stage of Estrus is E | 91 | N+ve | 2 | 2 | 2 | 1 |
Stage of Estrus is M | 91 | N+ve | 0 | 1 | 0 | 1 |
Stage of Estrus is D | 91 | N+ve | 5 | 5 | 7 | 4 |
Stage of Estrus is P | 91 | N+ve | 3 | 2 | 1 | 4 |
[p1] - T-Test: [G1RvsG4R]
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Stage of Estrus is E | 119 | N+ve | 1 | 1 |
Stage of Estrus is M | 119 | N+ve | 0 | 0 |
Stage of Estrus is P | 119 | N+ve | 3 | 2 |
Stage of Estrus is D | 119 | N+ve | 1 | 2 |
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Stage of Estrus is E | 119 | N+ve | 1 | 1 |
Stage of Estrus is M | 119 | N+ve | 0 | 0 |
Stage of Estrus is P | 119 | N+ve | 3 | 2 |
Stage of Estrus is D | 119 | N+ve | 1 | 2 |
Appendix 15, TABLE 1. Summary of Haematology and Coagulation Parameters on Day 91 – Males
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
RBC (10^12/L) | 91 [a] | Mean SD N %Diff | 9.26 0.36 10 - | 9.21 0.31 10 -0.57 | 9.28 0.41 10 0.21 | 9.38 0.34 10 1.26 |
HGB (g/L) | 91 [a] | Mean SD N %Diff | 158 6 10 - | 159 3 10 1 | 161 4 10 2 | 161 5 10 2 |
HCT (L/L) | 91 [a] | Mean SD N %Diff | 0.482 0.020 10 . | 0.495 0.016 10 2.782 | 0.496 0.015 10 3.073 | 0.516 * 0.019 10 7.081 |
MCV (fL) | 91 [a] | Mean SD N %Diff | 52.0 1.8 10 - | 53.8 1.2 10 3.4 | 53.5 1.5 10 2.9 | 55.0 * 2.0 10 5.8 |
MCH (pg) | 91 [a] | Mean SD N %Diff | 17.0 0.6 10 - | 17.3 0.5 10 1.8 | 17.4 0.5 10 2.1 | 17.2 0.8 10 0.9 |
MCHC (g/L) | 91 [a] | Mean SD N %Diff | 327 4 10 - | 322 8 10 -2 | 325 4 10 -1 | 312 * 8 10 -5 |
Retic A (10^12/L) | 91 [a] | Mean SD N %Diff | 0.170 0.027 10 - | 0.151 0.020 10 -11.019 | 0.152 0.024 10 -10.725 | 0.170 0.023 10 0.412 |
[a] - Anova & Dunnett: *: Significantly different from vehicle control group at p < 0.05
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Retic (%) | 91 [a] | Mean SD N %Diff | 1.84 0.30 10 - | 1.65 0.24 10 -10.45 | 1.64 0.30 10 -10.78 | 1.82 0.25 10 -0.98 |
RDW (%) | 91 [a] | Mean SD N %Diff | 12.3 0.5 10 - | 12.6 0.8 10 2.9 | 12.4 0.4 10 0.9 | 13.1 0.9 10 6.5 |
HDW (g/L) | 91 [a] | Mean SD N %Diff | 23.5 1.3 10 - | 24.1 2.1 10 2.3 | 24.0 1.4 10 2.2 | 24.7 1.6 10 5.0 |
Hyper (%) | 91 [a1] | Mean SD N %Diff | 0.6 0.3 10 . | 0.6 0.6 10 6.7 | 0.5 0.3 10 -10.0 | 0.3 0.2 10 -43.3 |
Hypo (%) | 91 [a1] | Mean SD N %Diff | 0.4 0.2 10 . | 0.8 1.1 10 92.9 | 0.6 0.4 10 42.9 | 4.3 * 4.8 10 911.9 |
Macro (%) | 91 [a1] | Mean SD N %Diff | 0.0 0.0 10 . | 0.0 0.1 10 . | 0.0 0.0 10 . | 0.0 0.0 10 . |
Micro (%) | 91 [a1] | Mean SD N %Diff | 0.0 0.0 10 . | 0.0 n 0.0 10 . | 0.0 n 0.0 10 . | 0.0 n 0.0 10 . |
[a] - Anova & Dunnett
[a1] - Anova & Dunnett(Rank): *: Significantly different from vehicle control group at p < 0.05; n - Inappropriate for statistics
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
RBC Frag- ments (10^12/L) | 91 [a] | Mean SD N %Diff | 0.14 0.02 10 - | 0.17 * 0.02 10 19.58 | 0.17 * 0.01 10 18.88 | 0.17 * 0.02 10 16.78 |
RBC Ghosts (10^12/L) | 91 [a] | Mean SD N %Diff | 0.08 0.01 10 - | 0.09 0.01 10 7.59 | 0.09 0.01 10 7.59 | 0.09 0.01 10 11.39 |
Plat (10^9/L) | 91 [a] | Mean SD N %Diff | 1614 109 10 - | 1766 * 138 10 9 | 1769 * 105 10 10 | 1710 173 10 6 |
MPV (fL) | 91 [a1] | Mean SD N %Diff | 11.2 0.4 10 - | 12.5 * 0.6 10 11.6 | 12.4 * 0.3 10 10.3 | 13.0 * 0.3 10 15.4 |
WBC (10^9/L) | 91 [a] | Mean SD N %Diff | 10.78 1.90 10 - | 12.01 2.60 10 11.38 | 10.12 1.62 10 -6.12 | 12.03 1.75 10 11.54 |
Neut A (10^9/L) | 91 [a] | Mean SD N %Diff | 3.22 0.98 10 - | 2.81 0.65 10 -12.72 | 2.67 0.65 10 -17.22 | 3.19 0.70 10 -0.96 |
Lymp A (10^9/L) | 91 [a] | Mean SD N %Diff | 6.81 0.99 10 - | 8.40 2.06 10 23.36 | 6.83 1.42 10 0.29 | 8.05 1.61 10 18.13 |
[a] - Anova & Dunnett: *: Significantly different from vehicle control group at p < 0.05
[a1] - Anova & Dunnett(Rank): *: Significantly different from vehicle control group at p < 0.05
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Mono A (10^9/L) | 91 [a] | Mean SD N %Diff | 0.41 0.10 10 - | 0.43 0.12 10 4.40 | 0.34 0.05 10 -16.14 | 0.43 0.11 10 4.40 |
Baso A (10^9/L) | 91 [a] | Mean SD N %Diff | 0.03 0.01 10 - | 0.03 0.02 10 6.25 | 0.02 0.01 10 -31.25 | 0.03 0.01 10 -6.25 |
Eosi A (10^9/L) | 91 [a1] | Mean SD N %Diff | 0.27 0.10 10 - | 0.29 0.10 10 7.55 | 0.23 0.07 10 -15.09 | 0.29 0.12 10 8.30 |
Prothrombin Time (seconds) | 91 [a2] | Mean SD N %Diff | 18.6 0.9 10 . | 18.6 1.1 10 -0.2 | 18.2 0.8 10 -2.1 | 18.6 1.1 10 0.0 |
APTT (seconds) | 91 [a] | Mean SD N %Diff | 9.0 1.9 10 - | 9.0 1.7 10 0.4 | 9.2 1.0 10 2.5 | 9.1 1.7 10 1.9 |
[a] - Anova & Dunnett
[a1] Anova & Dunnett(Log)
[a2] Anova & Dunnett(Rank)
Appendix 15, TABLE 2. Summary of Haematology and Coagulation Parameters on Day 119 – Males
Sex: Male Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
RBC (10^12/L) | 119 [p] | Mean SD N %Diff | 8.77 1.20 5 - | 9.16 0.25 5 4.47 |
HGB (g/L) | 119 [p] | Mean SD N %Diff | 147 21 5 - | 159 2 5 8 |
HCT (L/L) | 119 [p] | Mean SD N %Diff | 0.467 0.060 5 . | 0.500 0.007 5 6.889 |
MCV (fL) | 119 [p1] | Mean SD N %Diff | 53.4 0.9 5 - | 54.5 0.9 5 2.1 |
MCH (pg) | 119 [p1] | Mean SD N %Diff | 16.7 0.3 5 - | 17.4 * 0.4 5 3.8 |
MCHC (g/L) | 119 [p1] | Mean SD N %Diff | 313 8 5 - | 318 5 5 2 |
Retic A (10^12/L) | 119 [p2] | Mean SD N %Diff | 0.276 0.068 5 - | 0.202 * 0.019 5 -26.633 |
[p] - Wilcoxon(Rank): [G1RvsG4R]
[p1] - T-Test: [G1RvsG4R]
{MCH} G1RvsG4R: *: Significantly different from vehicle control group at p < 0.05
[p2] T-Test(Log): [G1RvsG4R]
G1RvsG4R: *: Significantly different from vehicle control group at p < 0.05
Sex: Male Da | y(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | |
Retic (%) | 119 [p] | Mean SD N %Diff | 3.29 1.43 5 - | 2.21 * 0.25 5 -32.68 |
RDW (%) | 119 [p] | Mean SD N %Diff | 15.3 3.9 5 - | 13.1 0.7 5 -14.6 |
HDW (g/L) | 119 [p1] | Mean SD N %Diff | 26.2 2.5 5 - | 25.2 1.7 5 -3.9 |
Hyper (%) | 119 [p1] | Mean SD N %Diff | 0.4 0.1 5 . | 0.5 0.2 5 25.0 |
Hypo (%) | 119 [p2] | Mean SD N %Diff | 2.7 2.2 5 . | 1.5 0.9 5 -44.5 |
Macro (%) | 119 [p] | Mean SD N %Diff | 0.6 1.3 5 . | 0.0 0.0 5 -100.0 |
Micro (%) | 119 [p] | Mean SD N %Diff | 0.0 0.0 5 . | 0.0 0.0 5 . |
[p] - Wilcoxon(Rank): [G1RvsG4R]
{Retic} G1RvsG4R: *: Significantly different from vehicle control group at p < 0.05
[p1] T-Test: [G1RvsG4R]
[p2] T-Test(Log): [G1RvsG4R]
Sex: Male Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
RBC Frag- ments (10^12/L) | 119 [p] | Mean SD N %Diff | 0.16 0.01 5 - | 0.16 0.01 5 0.00 |
RBC Ghosts (10^12/L) | 119 [p] | Mean SD N %Diff | 0.07 0.01 5 - | 0.09 * 0.01 5 27.78 |
Plat (10^9/L) | 119 [p] | Mean SD N %Diff | 1667 105 5 - | 1665 126 5 0 |
MPV (fL) | 119 [p] | Mean SD N %Diff | 11.4 0.6 5 - | 12.0 0.4 5 5.1 |
WBC (10^9/L) | 119 [p] | Mean SD N %Diff | 10.33 3.16 5 - | 9.22 0.90 5 -10.73 |
Neut A (10^9/L) | 119 [p] | Mean SD N %Diff | 4.06 2.46 5 - | 2.60 1.07 5 -35.89 |
Lymp A (10^9/L) | 119 [p] | Mean SD N %Diff | 5.65 1.16 5 - | 6.08 1.45 5 7.69 |
[p] - T-Test: [G1RvsG4R]
{RBC Ghosts} G1RvsG4R: *: Significantly different from vehicle control group at p < 0.05
Sex: Male Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Mono A (10^9/L) | 119 [p] | Mean SD N %Diff | 0.27 0.11 5 - | 0.28 0.06 5 2.22 |
Baso A (10^9/L) | 119 [p1] | Mean SD N %Diff | 0.02 0.02 5 - | 0.02 0.00 5 -8.33 |
Eosi A (10^9/L) | 119 [p2] | Mean SD N %Diff | 0.30 0.24 5 - | 0.20 0.06 5 -32.43 |
Prothrombin Time (seconds) | 119 [p1] | Mean SD N %Diff | 14.7 0.5 5 . | 15.6 * 0.3 5 5.8 |
APTT (seconds) | 119 [p] | Mean SD N %Diff | 7.5 0.9 5 - | 9.0 * 0.8 5 20.6 |
[p] - T-Test: [G1RvsG4R]
{APTT} G1RvsG4R: *: Significantly different from vehicle control group at p < 0.05
[p1] - Wilcoxon(Rank): [G1RvsG4R]
{Prothrombin} G1RvsG4R: *: Significantly different from vehicle control group at p < 0.05 [p2] T-Test(Log): [G1RvsG4R]
Appendix 15, TABLE 3. Summary of Haematology and Coagulation Parameters on Day 91 - Females
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
RBC (10^12/L) | 91 [a] | Mean SD N %Diff | 8.64 0.37 10 - | 8.83 0.31 10 2.23 | 8.65 0.30 10 0.16 | 8.45 0.34 10 -2.13 |
HGB (g/L) | 91 [a] | Mean SD N %Diff | 156 6 10 - | 158 4 10 1 | 158 5 10 1 | 156 5 10 0 |
HCT (L/L) | 91 [a] | Mean SD N %Diff | 0.473 0.016 10 . | 0.490 0.012 10 3.443 | 0.489 0.016 10 3.338 | 0.494 * 0.020 10 4.246 |
MCV (fL) | 91 [a] | Mean SD N %Diff | 54.8 1.3 10 - | 55.5 1.7 10 1.2 | 56.6 1.9 10 3.1 | 58.5 * 2.4 10 6.6 |
MCH (pg) | 91 [a] | Mean SD N %Diff | 18.1 0.5 10 - | 18.0 0.6 10 -0.9 | 18.3 0.7 10 1.0 | 18.4 0.7 10 1.8 |
MCHC (g/L) | 91 [a] | Mean SD N %Diff | 330 3 10 - | 323 * 3 10 -2 | 323 * 5 10 -2 | 316 * 7 10 -4 |
Retic A (10^12/L) | 91 [a] | Mean SD N %Diff | 0.182 0.042 10 - | 0.140 0.030 10 -22.984 | 0.168 0.044 10 -8.009 | 0.170 0.046 10 -6.967 |
[a] - Anova & Dunnett: *: Significantly different from vehicle control group at p < 0.05
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Retic (%) | 91 [a] | Mean SD N %Diff | 2.13 0.54 10 - | 1.59 0.33 10 -25.18 | 1.95 0.54 10 -8.33 | 2.02 0.58 10 -5.04 |
RDW (%) | 91 [a] | Mean SD N %Diff | 11.2 0.5 10 - | 11.5 0.5 10 2.6 | 11.3 0.5 10 1.3 | 11.6 0.3 10 3.9 |
HDW (g/L) | 91 [a] | Mean SD N %Diff | 20.2 1.1 10 - | 20.2 0.8 10 -0.2 | 19.9 1.0 10 -1.4 | 20.8 0.6 10 2.7 |
Hyper (%) | 91 [a1] | Mean SD N %Diff | 0.3 0.2 10 . | 0.2 0.1 10 -50.0 | 0.1 * 0.1 10 -66.7 | 0.1 * 0.2 10 -70.0 |
Hypo (%) | 91 [a1] | Mean SD N %Diff | 0.2 0.1 10 . | 0.3 0.2 10 17.4 | 0.2 0.1 10 -8.7 | 1.8 * 1.4 10 700.0 |
Macro (%) | 91 [a1] | Mean SD N %Diff | 0.0 0.0 10 . | 0.0 n 0.0 10 . | 0.0 n 0.0 10 . | 0.0 n 0.0 10 . |
Micro (%) | 91 [a1] | Mean SD N %Diff | 0.0 0.0 10 . | 0.0 n 0.0 10 . | 0.0 n 0.0 10 . | 0.0 n 0.0 10 . |
[a] - Anova & Dunnett
[a1] - Anova & Dunnett(Rank): *: Significantly different from vehicle control group at p < 0.05; n - Inappropriate for statistics
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
RBC Frag- ments (10^12/L) | 91 [a] | Mean SD N %Diff | 0.16 0.01 10 - | 0.17 0.01 10 8.39 | 0.17 0.01 10 6.45 | 0.16 0.01 10 5.81 |
RBC Ghosts (10^12/L) | 91 [a] | Mean SD N %Diff | 0.09 0.01 10 - | 0.10 0.01 10 2.13 | 0.10 0.01 10 2.13 | 0.10 0.01 10 9.57 |
Plat (10^9/L) | 91 [a] | Mean SD N %Diff | 1690 86 10 - | 1819 * 123 10 8 | 1847 * 110 10 9 | 1914 * 113 10 13 |
MPV (fL) | 91 [a] | Mean SD N %Diff | 11.8 0.5 10 - | 12.6 * 0.6 10 6.9 | 13.0 * 0.6 10 10.0 | 12.8 * 0.5 10 9.0 |
WBC (10^9/L) | 91 [a1] | Mean SD N %Diff | 6.42 1.31 10 - | 6.51 1.12 10 1.37 | 6.36 1.06 10 -0.83 | 6.75 2.64 10 5.19 |
Neut A (10^9/L) | 91 [a] | Mean SD N %Diff | 1.60 0.44 10 - | 1.53 0.40 10 -4.68 | 1.51 0.26 10 -5.74 | 1.46 0.43 10 -8.98 |
Lymp A (10^9/L) | 91 [a1] | Mean SD N %Diff | 4.44 1.16 10 - | 4.55 0.83 10 2.48 | 4.49 0.90 10 1.26 | 4.97 2.25 10 11.97 |
[a] - Anova & Dunnett: *: Significantly different from vehicle control group at p < 0.05 [a1] Anova & Dunnett(Rank)
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Mono A (10^9/L) | 91 [a] | Mean SD N %Diff | 0.19 0.09 10 - | 0.19 0.08 10 -2.58 | 0.18 0.06 10 -9.79 | 0.15 0.06 10 -21.13 |
Baso A (10^9/L) | 91 [a1] | Mean SD N %Diff | 0.01 0.01 10 - | 0.01 0.00 10 -21.43 | 0.01 0.00 10 -50.00 | 0.01 0.01 10 -14.29 |
Eosi A (10^9/L) | 91 [a] | Mean SD N %Diff | 0.14 0.05 10 - | 0.20 0.10 10 42.55 | 0.15 0.05 10 8.51 | 0.14 0.06 10 -3.55 |
Prothrombin Time (seconds) | 91 [a2] | Mean SD N %Diff | 19.8 1.0 10 . | 17.8 * 1.2 10 -10.2 | 18.4 * 0.7 10 -7.3 | 19.0 1.2 10 -4.1 |
APTT (seconds) | 91 [a] | Mean SD N %Diff | 10.1 1.4 10 - | 9.9 2.7 10 -2.1 | 8.6 1.1 10 -15.1 | 9.3 1.1 10 -7.3 |
[a] - Anova & Dunnett(Log)
[a1] - Anova & Dunnett(Rank)
[a2] Anova & Dunnett: *: Significantly different from vehicle control group at p < 0.05
Appendix 15, TABLE 4. Summary of Haematology and Coagulation Parameters on Day 119 – Females
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
RBC (10^12/L) | 119 [p] | Mean SD N %Diff | 8.50 0.30 5 - | 8.44 0.46 5 -0.80 |
HGB (g/L) | 119 [p1] | Mean SD N %Diff | 152 7 5 - | 154 3 5 2 |
HCT (L/L) | 119 [p] | Mean SD N %Diff | 0.476 0.018 5 . | 0.482 0.009 5 1.260 |
MCV (fL) | 119 [p] | Mean SD N %Diff | 56.0 2.4 5 - | 57.3 3.0 5 2.2 |
MCH (pg) | 119 [p] | Mean SD N %Diff | 17.9 0.8 5 - | 18.3 1.0 5 2.5 |
MCHC (g/L) | 119 [p] | Mean SD N %Diff | 319 4 5 - | 320 3 5 0 |
Retic A (10^12/L) | 119 [p] | Mean SD N %Diff | 0.195 0.030 5 - | 0.200 0.042 5 2.669 |
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Retic (%) | 119 [p] | Mean SD N %Diff | 2.29 0.37 5 - | 2.36 0.44 5 3.05 |
RDW (%) | 119 [p] | Mean SD N %Diff | 11.9 0.4 5 - | 11.8 0.7 5 -1.3 |
HDW (g/L) | 119 [p] | Mean SD N %Diff | 20.0 0.9 5 - | 21.0 2.0 5 5.0 |
Hyper (%) | 119 [p] | Mean SD N %Diff | 0.1 0.1 5 . | 0.2 0.2 5 175.0 |
Hypo (%) | 119 [p] | Mean SD N %Diff | 0.7 0.3 5 . | 0.8 0.6 5 13.5 |
Macro (%) | 119 [p1] | Mean SD N %Diff | 0.0 0.0 5 . | 0.0 0.0 5 . |
Micro (%) | 119 [p1] | Mean SD N %Diff | 0.0 0.0 5 . | 0.0 0.0 5 . |
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
RBC Frag- ments (10^12/L) | 119 [p] | Mean SD N %Diff | 0.16 0.01 5 - | 0.15 0.03 5 -2.56 |
RBC Ghosts (10^12/L) | 119 [p] | Mean SD N %Diff | 0.09 0.02 5 - | 0.10 0.02 5 6.38 |
Plat (10^9/L) | 119 [p] | Mean SD N %Diff | 1699 63 5 - | 1675 138 5 -1 |
MPV (fL) | 119 [p] | Mean SD N %Diff | 12.5 0.6 5 - | 12.6 0.7 5 0.8 |
WBC (10^9/L) | 119 [p] | Mean SD N %Diff | 4.81 0.71 5 - | 5.34 2.13 5 11.01 |
Neut A (10^9/L) | 119 [p1] | Mean SD N %Diff | 1.26 0.18 5 - | 1.51 0.69 5 19.30 |
Lymp A (10^9/L) | 119 [p] | Mean SD N %Diff | 3.29 0.82 5 - | 3.52 1.54 5 7.18 |
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Mono A (10^9/L) | 119 [p] | Mean SD N %Diff | 0.14 0.06 5 - | 0.10 0.04 5 -30.99 |
Baso A (10^9/L) | 119 [p] | Mean SD N %Diff | 0.00 0.01 5 - | 0.01 0.01 5 150.00 |
Eosi A (10^9/L) | 119 [p] | Mean SD N %Diff | 0.09 0.03 5 - | 0.19 0.14 5 104.35 |
Prothrombin Time (seconds) | 119 [p] | Mean SD N %Diff | 16.0 1.4 5 . | 16.4 0.5 5 2.2 |
APTT (seconds) | 119 [p] | Mean SD N %Diff | 10.7 1.9 5 - | 8.2 * 0.8 5 -23.6 |
T-Test: [G1RvsG4R]
{APTT} G1RvsG4R: *: Significantly different from vehicle control group at p < 0.05
Appendix 15, TABLE 5. Summary of Clinical Chemistry Parameters on Day 91 – Males
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Glu (mmol/L) | 91 [a] | Mean SD N %Diff | 6.67 0.30 10 . | 6.29 0.57 10 -5.61 | 6.56 0.45 10 -1.61 | 6.24 0.49 10 -6.42 |
BUN (mmol/L) | 91 [a] | Mean SD N %Diff | 5.09 0.81 10 . | 5.34 0.72 10 4.91 | 4.93 0.70 10 -3.30 | 5.17 0.61 10 1.43 |
Creat (µmol/L) | 91 [a] | Mean SD N %Diff | 26 6 10 - | 24 4 10 -6 | 29 3 10 11 | 24 3 10 -7 |
AST (U/L) | 91 [a] | Mean SD N %Diff | 93 8 10 . | 93 9 10 0 | 94 10 10 2 | 92 9 10 -1 |
ALT (U/L) | 91 [a] | Mean SD N %Diff | 32 6 10 . | 38 5 10 20 | 34 8 10 9 | 38 10 10 21 |
GGT (U/L) | 91 [a] | Mean SD N %Diff | 3 1 10 . | 3 1 10 19 | 3 1 10 12 | 3 1 10 0 |
Alp (U/L) | 91 [a] | Mean SD N %Diff | 71 10 10 . | 63 11 10 -11 | 61 11 10 -14 | 75 15 10 7 |
[a] - Anova & Dunnett
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
T.Bil (µmol/L) | 91 [a] | Mean SD N %Diff | 2.71 0.61 10 . | 2.62 0.95 10 -3.03 | 1.99 * 0.31 10 -26.36 | 2.17 0.35 10 -19.70 |
T.Chol (mmol/L) | 91 [a1] | Mean SD N %Diff | 2.34 0.45 10 . | 2.39 0.38 10 2.09 | 2.36 0.40 10 0.98 | 2.25 0.42 10 -3.72 |
HDL Cholesterol (mmol/L) | 91 [a1] | Mean SD N %Diff | 2.11 0.42 10 . | 2.01 0.33 10 -4.60 | 2.05 0.35 10 -2.61 | 2.09 0.46 10 -0.95 |
LDL Cholesterol (mmol/L) | 91 [a1] | Mean SD N %Diff | 0.04 0.12 10 . | 0.18 * 0.10 10 413.64 | 0.12 0.10 10 238.07 | -0.06 0.14 10 -265.91 |
Trig (mmol/L) | 91 [a2] | Mean SD N %Diff | 0.98 0.38 10 . | 0.99 0.35 10 0.20 | 0.96 0.42 10 -2.95 | 1.12 0.42 10 13.52 |
T.Pro (g/L) | 91 [a1] | Mean SD N %Diff | 68.3 1.3 10 - | 67.8 2.0 10 -0.7 | 68.8 2.1 10 0.7 | 67.8 1.7 10 -0.7 |
ALB (g/L) | 91 [a1] | Mean SD N %Diff | 34.4 1.0 10 - | 34.0 1.6 10 -1.2 | 34.4 0.9 10 0.1 | 34.2 1.1 10 -0.8 |
[a] - Anova & Dunnett(Rank): *: Significantly different from vehicle control group at p < 0.05
[a1] - Anova & Dunnett: *: Significantly different from vehicle control group at p < 0.05 [a2] - Anova & Dunnett(Log)
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
GLOB (g/L) | 91 [a] | Mean SD N %Diff | 33.8 1.4 10 - | 33.8 0.9 10 -0.3 | 34.3 1.5 10 1.4 | 33.7 1.2 10 -0.6 |
A/G (ratio) | 91 [a] | Mean SD N %Diff | 1.02 0.06 10 - | 1.01 0.05 10 -1.09 | 1.00 0.04 10 -1.42 | 1.02 0.05 10 -0.26 |
Pi (mmol/L) | 91 [a1] | Mean SD N %Diff | 1.85 0.17 10 . | 1.93 0.14 10 4.82 | 1.78 0.17 10 -3.79 | 1.94 0.12 10 5.37 |
Ca (mmol/L) | 91 [a] | Mean SD N %Diff | 2.90 0.08 10 . | 2.81 0.13 10 -2.83 | 2.49 * 0.12 10 -13.88 | 2.86 0.06 10 -1.14 |
Na (mEq/L) | 91 [a] | Mean SD N %Diff | 146.2 0.9 10 - | 146.8 0.8 10 0.4 | 147.6 * 1.4 10 1.0 | 146.7 1.1 10 0.4 |
K (mEq/L) | 91 [a1] | Mean SD N %Diff | 3.74 0.22 10 - | 3.94 0.21 10 5.24 | 4.08 * 0.30 10 9.00 | 3.91 0.25 10 4.49 |
Cl (mEq/L) | 91 [a] | Mean SD N %Diff | 95.7 0.8 10 - | 95.8 1.1 10 0.2 | 96.0 0.9 10 0.4 | 95.7 1.0 10 0.1 |
[a] - Anova & Dunnett: *: Significantly different from vehicle control group at p < 0.05
[a1] - Anova & Dunnett(Rank): *: Significantly different from vehicle control group at p < 0.05
Appendix 15, TABLE 6. Summary of Clinical Chemistry Parameters on Day 119 – Males
Sex: Male Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Glu (mmol/L) | 119 [p] | Mean SD N %Diff | 8.29 0.51 5 . | 6.66 * 0.63 5 -19.69 |
BUN (mmol/L) | 119 [p] | Mean SD N %Diff | 5.92 0.98 5 . | 5.62 0.47 5 -5.00 |
Creat (µmol/L) | 119 [p] | Mean SD N %Diff | 36 6 5 - | 36 5 5 -1 |
AST (U/L) | 119 [p1] | Mean SD N %Diff | 177 117 5 . | 99 * 13 5 -44 |
ALT (U/L) | 119 [p1] | Mean SD N %Diff | 109 127 5 . | 35 * 3 5 -68 |
GGT (U/L) | 119 [p] | Mean SD N %Diff | 3 1 5 . | 2 0 5 -21 |
Alp (U/L) | 119 [p] | Mean SD N %Diff | 71 15 5 . | 63 22 5 -11 |
[p] - T-Test: [G1RvsG4R]
{Glu} G1RvsG4R: *: Significantly different from vehicle control group at p < 0.05
[p1] - Wilcoxon(Rank): [G1RvsG4R]
{AST} G1RvsG4R: *: Significantly different from vehicle control group at p < 0.05
{ALT} G1RvsG4R: *: Significantly different from vehicle control group at p < 0.05
Sex: Male Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
T.Bil (µmol/L) | 119 [p] | Mean SD N %Diff | 2.15 0.58 5 . | 1.84 0.39 5 -14.33 |
T.Chol (mmol/L) | 119 [p] | Mean SD N %Diff | 2.99 0.42 5 . | 2.82 0.59 5 -5.43 |
HDL Cholesterol (mmol/L) | 119 [p] | Mean SD N %Diff | 2.43 0.36 5 . | 2.25 0.54 5 -7.34 |
LDL Cholesterol (mmol/L) | 119 [p] | Mean SD N %Diff | 0.41 0.12 5 . | 0.45 0.09 5 8.03 |
Trig (mmol/L) | 119 [p] | Mean SD N %Diff | 0.73 0.26 5 . | 0.65 0.16 5 -11.72 |
T.Pro (g/L) | 119 [p] | Mean SD N %Diff | 67.3 2.0 5 - | 67.0 0.5 5 -0.5 |
ALB (g/L) | 119 [p] | Mean SD N %Diff | 30.5 1.4 5 - | 30.5 1.8 5 -0.2 |
[p] - T-Test: [G1RvsG4R]
Sex: Male Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
GLOB (g/L) | 119 [p] | Mean SD N %Diff | 36.8 1.3 5 - | 36.5 2.1 5 -0.8 |
A/G (ratio) | 119 [p] | Mean SD N %Diff | 0.83 0.05 5 - | 0.84 0.10 5 1.00 |
Pi (mmol/L) | 119 [p] | Mean SD N %Diff | 1.85 0.11 5 . | 1.88 0.14 5 1.19 |
Ca (mmol/L) | 119 [p] | Mean SD N %Diff | 2.91 0.03 5 . | 2.83 * 0.04 5 -2.61 |
Na (mEq/L) | 119 [p] | Mean SD N %Diff | 145.8 1.0 5 - | 146.3 1.8 5 0.3 |
K (mEq/L) | 119 [p] | Mean SD N %Diff | 3.88 0.34 5 - | 3.81 0.43 5 -1.70 |
Cl (mEq/L) | 119 [p] | Mean SD N %Diff | 97.4 1.4 5 - | 97.4 0.6 5 0.0 |
[p] - T-Test: [G1RvsG4R]
{Ca} G1RvsG4R: *: Significantly different from vehicle control group at p < 0.05
Appendix 15, TABLE 7. Summary of Clinical Chemistry Parameters on Day 91 – Females
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Glu (mmol/L) | 91 [a] | Mean SD N %Diff | 6.41 0.59 10 . | 6.35 0.46 10 -1.00 | 6.69 0.84 10 4.34 | 6.46 0.80 10 0.80 |
BUN (mmol/L) | 91 [a] | Mean SD N %Diff | 5.38 1.06 10 . | 5.75 0.89 10 6.97 | 5.43 0.85 10 0.87 | 5.61 0.72 10 4.37 |
Creat (µmol/L) | 91 [a] | Mean SD N %Diff | 36 5 10 - | 32 4 10 -12 | 34 4 10 -5 | 30 * 3 10 -19 |
AST (U/L) | 91 [a1] | Mean SD N %Diff | 102 14 10 . | 142 76 10 40 | 100 20 10 -1 | 97 10 10 -5 |
ALT (U/L) | 91 [a1] | Mean SD N %Diff | 31 6 10 . | 50 54 10 60 | 29 9 10 -9 | 27 6 10 -14 |
GGT (U/L) | 91 [a1] | Mean SD N %Diff | 2 0 10 . | 3 0 10 17 | 2 0 10 0 | 3 1 10 17 |
Alp (U/L) | 91 [a] | Mean SD N %Diff | 38 4 10 . | 32 * 7 10 -16 | 37 5 10 -3 | 34 4 10 -10 |
[a] - Anova & Dunnett: *: Significantly different from vehicle control group at p < 0.05 [a1] - Anova & Dunnett(Rank)
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
T.Bil (µmol/L) | 91 [a] | Mean SD N %Diff | 2.77 1.13 10 . | 2.38 0.54 10 -14.06 | 2.73 0.66 10 -1.23 | 2.63 0.55 10 -5.13 |
T.Chol (mmol/L) | 91 [a] | Mean SD N %Diff | 2.25 0.32 10 . | 2.36 0.52 10 4.66 | 2.48 0.36 10 10.04 | 2.38 0.32 10 5.51 |
HDL Cholesterol (mmol/L) | 91 [a] | Mean SD N %Diff | 1.70 0.23 10 . | 2.04 0.44 10 20.09 | 2.04 0.26 10 19.98 | 1.94 0.25 10 14.50 |
LDL Cholesterol (mmol/L) | 91 [a1] | Mean SD N %Diff | 0.46 0.13 10 . | 0.18 * 0.14 10 -59.51 | 0.32 0.16 10 -29.23 | 0.33 0.18 10 -28.75 |
Trig (mmol/L) | 91 [a1] | Mean SD N %Diff | 0.49 0.08 10 . | 0.67 0.21 10 36.11 | 0.60 0.23 10 20.69 | 0.54 0.13 10 9.33 |
T.Pro (g/L) | 91 [a] | Mean SD N %Diff | 68.7 3.5 10 - | 70.0 2.9 10 1.9 | 70.0 1.8 10 1.8 | 69.8 2.7 10 1.5 |
ALB (g/L) | 91 [a2] | Mean SD N %Diff | 36.6 3.0 10 - | 37.8 2.8 10 3.1 | 38.1 1.8 10 3.9 | 37.4 1.7 10 2.1 |
[a] - Anova & Dunnett
[a1] - Anova & Dunnett(Log): *: Significantly different from vehicle control group at p < 0.05 [a2] - Anova & Dunnett(Rank)
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
GLOB (g/L) | 91 [a] | Mean SD N %Diff | 32.1 1.3 10 - | 32.2 1.3 10 0.4 | 31.9 0.9 10 -0.6 | 32.4 1.5 10 0.8 |
A/G (ratio) | 91 [a] | Mean SD N %Diff | 1.14 0.10 10 - | 1.18 0.11 10 2.81 | 1.20 0.07 10 4.56 | 1.16 0.05 10 1.28 |
Pi (mmol/L) | 91 [a] | Mean SD N %Diff | 1.75 0.19 10 . | 1.65 0.17 10 -5.66 | 1.63 0.15 10 -7.14 | 1.74 0.11 10 -0.51 |
Ca (mmol/L) | 91 [a] | Mean SD N %Diff | 2.80 0.11 10 . | 2.79 0.10 10 -0.43 | 2.71 0.16 10 -3.39 | 2.87 0.08 10 2.32 |
Na (mEq/L) | 91 [a] | Mean SD N %Diff | 143.2 1.6 10 - | 144.7 * 1.5 10 1.1 | 145.1 * 1.3 10 1.3 | 144.4 0.9 10 0.9 |
K (mEq/L) | 91 [a] | Mean SD N %Diff | 3.53 0.40 10 - | 3.98 * 0.39 10 12.84 | 3.81 0.25 10 7.88 | 3.56 0.26 10 0.82 |
Cl (mEq/L) | 91 [a1] | Mean SD N %Diff | 95.1 0.9 10 - | 95.3 1.3 10 0.2 | 95.5 0.8 10 0.4 | 95.6 0.7 10 0.5 |
[a] - Anova & Dunnett: *: Significantly different from vehicle control group at p < 0.05 [a1] - Anova & Dunnett(Rank)
Appendix 15, TABLE 8. Summary of Clinical Chemistry Parameters on Day 119 – Females
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Glu (mmol/L) | 119 [p] | Mean SD N %Diff | 7.62 0.81 5 . | 7.75 0.53 5 1.65 |
BUN (mmol/L) | 119 [p] | Mean SD N %Diff | 4.98 0.68 5 . | 5.89 1.07 5 18.32 |
Creat (µmol/L) | 119 [p] | Mean SD N %Diff | 44 3 5 - | 49 8 5 11 |
AST (U/L) | 119 [p1] | Mean SD N %Diff | 122 52 5 . | 126 7 5 4 |
ALT (U/L) | 119 [p] | Mean SD N %Diff | 33 7 5 . | 42 13 5 27 |
GGT (U/L) | 119 [p] | Mean SD N %Diff | 2 0 5 . | 2 1 5 22 |
Alp (U/L) | 119 [p] | Mean SD N %Diff | 27 7 5 . | 33 9 5 21 |
[p] - T-Test: [G1RvsG4R]
[p1] - Wilcoxon(Rank): [G1RvsG4R]
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
T.Bil (µmol/L) | 119 [p] | Mean SD N %Diff | 1.96 0.89 5 . | 2.14 0.49 5 9.61 |
T.Chol (mmol/L) | 119 [p] | Mean SD N %Diff | 2.75 0.30 5 . | 2.62 0.28 5 -4.59 |
HDL Cholesterol (mmol/L) | 119 [p] | Mean SD N %Diff | 2.10 0.24 5 . | 2.08 0.22 5 -0.67 |
LDL Cholesterol (mmol/L) | 119 [p] | Mean SD N %Diff | 0.54 0.09 5 . | 0.43 0.08 5 -19.66 |
Trig (mmol/L) | 119 [p] | Mean SD N %Diff | 0.55 0.12 5 . | 0.52 0.15 5 -5.42 |
T.Pro (g/L) | 119 [p] | Mean SD N %Diff | 72.6 3.4 5 - | 70.5 3.6 5 -2.9 |
ALB (g/L) | 119 [p] | Mean SD N %Diff | 40.1 2.8 5 - | 38.3 4.2 5 -4.4 |
[p] - T-Test: [G1RvsG4R]
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
GLOB (g/L) | 119 [p] | Mean SD N %Diff | 32.5 0.9 5 - | 32.2 1.4 5 -0.9 |
A/G (ratio) | 119 [p] | Mean SD N %Diff | 1.23 0.07 5 - | 1.19 0.17 5 -3.17 |
Pi (mmol/L) | 119 [p] | Mean SD N %Diff | 1.47 0.14 5 . | 1.38 0.24 5 -6.27 |
Ca (mmol/L) | 119 [p] | Mean SD N %Diff | 2.86 0.12 5 . | 2.86 0.13 5 0.07 |
Na (mEq/L) | 119 [p] | Mean SD N %Diff | 143.5 1.4 5 - | 146.2 * 1.1 5 1.9 |
K (mEq/L) | 119 [p] | Mean SD N %Diff | 3.73 0.64 5 - | 3.26 0.47 5 -12.70 |
Cl (mEq/L) | 119 [p] | Mean SD N %Diff | 95.5 1.5 5 - | 96.3 1.5 5 0.8 |
[p] - T-Test: [G1RvsG4R]
{Na} G1RvsG4R: *: Significantly different from vehicle control group at p < 0.05
Appendix 15, TABLE 9. Summary of Thyroid Hormone Profile on Day 91 – Males
Sex: Male Da | y(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | |
TSH (ng/mL) | 91 [a] | Mean SD N %Diff | 0.62 0.30 9a . | 0.64 0.47 9a 1.78 | 0.70 0.29 9a 12.28 | 0.63 0.37 10 1.21 |
T3 (ng/mL) | 91 [a1] | Mean SD N %Diff | 0.35 0.07 10 . | 0.39 0.04 10 11.53 | 0.37 0.05 10 5.48 | 0.36 0.04 10 4.90 |
T4 (ng/mL) | 91 [a1] | Mean SD N %Diff | 16.36 3.61 10 . | 17.44 3.43 10 6.60 | 16.85 2.17 10 2.96 | 16.95 2.51 10 3.58 |
a: Values below LLOQ (Lower Limit of Quantification for TSH is below 0.1 ng/mL) were not considered for analysis
[a] - Anova & Dunnett(Log)
[a1] Anova & Dunnett
Page 231/475
Appendix 15, TABLE 10. Summary of Thyroid Hormone Profile on Day 119 – Males
Sex: Male Da | y(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | |
TSH (ng/mL) | 119 [p] | Mean SD N %Diff | 1.17 0.26 5 . | 1.15 0.60 5 -2.05 |
T3 (ng/mL) | 119 [p1] | Mean SD N %Diff | 0.56 0.04 5 . | 0.53 0.11 5 -6.05 |
T4 (ng/mL) | 119 [p] | Mean SD N %Diff | 21.39 4.58 5 . | 19.95 8.81 5 -6.73 |
[p] - T-Test: [G1RvsG4R]
[p1] Wilcoxon(Rank): [G1RvsG4R]
Appendix 15, TABLE 11. Summary of Thyroid Hormone Profile on Day 91 – Females
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
TSH (ng/mL) | 91 [a] | Mean SD N %Diff | 0.53 0.26 7a . | 0.48 0.24 7a -8.70 | 0.47 0.19 9a -11.02 | 0.53 0.24 9a 1.24 |
T3 (ng/mL) | 91 [a1] | Mean SD N %Diff | 0.33 0.06 10 . | 0.36 0.06 10 8.87 | 0.36 0.06 10 9.79 | 0.36 0.04 9a 10.77 |
T4 (ng/mL) | 91 [a] | Mean SD N %Diff | 10.85 2.14 10 . | 11.87 2.57 10 9.40 | 10.80 2.57 10 -0.43 | 10.01 2.42 10 -7.76 |
a: Values below LLOQ (Lower Limit of Quantification for TSH is below 0.1 ng/mL and for T3 is below 0.2 ng/mL) were not considered for analysis
Appendix 15, TABLE 12. Summary of Thyroid Hormone Profile on Day 119 – Females
Sex: Female Da | y(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | |
TSH (ng/mL) | 119 [p] | Mean SD N %Diff | 0.91 0.67 4a . | 0.86 0.31 5 -6.19 |
T3 (ng/mL) | 119 [p1] | Mean SD N %Diff | 0.49 0.13 5 . | 0.51 0.08 4a 5.02 |
T4 (ng/mL) | 119 [p] | Mean SD N %Diff | 11.78 4.38 5 . | 13.06 5.05 5 10.88 |
a: Values below LLOQ (Lower Limit of Quantification for TSH is below 0.1 ng/mL and for T3 is below 0.2 ng/mL) were not considered for analysis
[p] - T-Test: [G1RvsG4R]
[p1] Wilcoxon(Rank): [G1RvsG4R]
Appendix 15, TABLE 13. Summary of Clinical Analysis of Urine on Day 91 – Males
Group No. | G1 | G2 | G3 | G4 |
Dose (mg/kg/day) | 0 | 111 | 333 | 1000 |
No. of rats | 10 | 10 | 10 | 10 |
Volume (mL) 1 | 27.5
| 38.2
| 29.0
| 34.8
|
Glucose2 | 0
| 0
| 0
| 0
|
Bilirubin2 | 0
| 0
| 0
| 0
|
Ketone Bodies2 | 0
| 0
| 0
| 1
|
Specific gravity 1 | 1.012 | 1.009 | 1.012
| 1.011
|
pH 1 |
9.0 |
8.9 | 9.0
| 9.0
|
Proteins2 |
5 |
4 | 7
| 7
|
Urobilinogen2,3 |
0 |
0 | 0
| 0
|
Nitrite2 |
0 |
2 | 0
| 0
|
Erythrocytes2 Ery/µL |
1 |
0 | 0
| 0
|
Leukocytes2 Leu/µL |
7 |
6 | 8 | 8 |
1: Mean value 2: Incidences of findings
3: Values less than or equal to 3.2 are normal and hence not considered as positive findings/incidence.
Appendix 15, TABLE 14. Summary of Clinical Analysis of Urine on Day 119 – Males
Group No. | G1R | G4R |
Dose (mg/kg/day) | 0 | 1000 |
No. of rats | 5 | 5 |
Volume (mL) 1 | 12.9
| 17.4
|
Glucose2 | 0
| 0
|
Bilirubin2 | 0
| 0
|
Ketone Bodies2 | 3
| 1
|
Specific gravity 1 | 1.024
| 1.024
|
pH 1 | 9.0
| 8.9
|
Proteins2 | 4
| 5
|
Urobilinogen2,3 | 0
| 2
|
Nitrite2 | 2
| 1
|
Erythrocytes2 Ery/µL | 0
| 0
|
Leukocytes2 Leu/µL | 4 | 5 |
1: Mean value 2: Incidences of findings
3: Values less than or equal to 3.2 are normal and hence not considered as positive findings/incidence.
Appendix 15, TABLE 15. Summary of Clinical Analysis of Urine on Day 91 – Females
Group No. | G1 | G2 | G3 | G4 |
Dose (mg/kg/day) | 0 | 111 | 333 | 1000 |
No. of rats | 10 | 10 | 10 | 10 |
Volume (mL) 1 | 24.6
| 20.5
| 17.8
| 15.7
|
Glucose2 | 0
| 0
| 0
| 0
|
Bilirubin2 | 0
| 0
| 0
| 0
|
Ketone Bodies2 | 0
| 0
| 0
| 0
|
Specific gravity 1 | 1.011 | 1.011 | 1.014
| 1.016
|
pH 1 |
9.0 |
9.0 | 8.9
| 9.0
|
Proteins2 |
3 |
4 | 7
| 7
|
Urobilinogen2,3 |
0 |
0 | 0
| 0
|
Nitrite2 |
1 |
2 | 2
| 2
|
Erythrocytes2 Ery/µL |
0 |
0 | 0
| 0
|
Leukocytes2 Leu/µL |
4 |
4 | 3 | 4 |
1: Mean value 2: Incidences of findings
3: Values less than or equal to 3.2 are normal and hence not considered as positive findings/incidence.
Appendix 15, TABLE 16. Summary of Clinical Analysis of Urine on Day 119 – Females
Group No. | G1R | G4R |
Dose (mg/kg/day) | 0 | 1000 |
No. of rats | 5 | 5 |
Volume (mL) 1 | 17.0
| 11.2
|
Glucose2 | 0
| 0
|
Bilirubin2 | 0
| 2
|
Ketone Bodies2 | 0
| 0
|
Specific gravity 1 | 1.017
| 1.024
|
pH 1 | 9.0
| 9.0
|
Proteins2 | 4
| 5
|
Urobilinogen2,3 | 2
| 3
|
Nitrite2 | 1
| 1
|
Erythrocytes2 Ery/µL | 0
| 1
|
Leukocytes2 Leu/µL | 3 | 5 |
1: Mean value 2: Incidences of findings
3: Values less than or equal to 3.2 are normal and hence not considered as positive findings/incidence.
Appendix 15, TABLE 17. Summary of Terminal Fasting Body Weights and Organ Weights on Day 91 – Males
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Terminal Fasting BW (g) | 91 [a] | Mean SD N %Diff | 509.88 49.34 10 - | 478.23 55.04 10 -6.21 | 491.21 37.99 10 -3.66 | 495.90 38.71 10 -2.74 |
Adrenals (g) | 91 [a] | Mean SD N %Diff | 0.0759 0.0101 10 - | 0.0686 0.0101 10 -9.6719 | 0.0693 0.0102 10 -8.6441 | 0.0738 0.0146 10 -2.7540 |
Brain (g) | 91 [a] | Mean SD N %Diff | 2.1871 0.0848 10 - | 2.1804 0.0952 10 -0.3050 | 2.2186 0.0756 10 1.4398 | 2.1833 0.0823 10 -0.1747 |
Epididymides (g) | 91 [a] | Mean SD N %Diff | 1.6373 0.1344 10 - | 1.5911 0.1162 10 -2.8217 | 1.5410 0.1319 10 -5.8852 | 1.6232 0.1804 10 -0.8630 |
Heart (g) | 91 [a] | Mean SD N %Diff | 1.4513 0.1114 10 - | 1.4222 0.1513 10 -2.0065 | 1.4791 0.1170 10 1.9169 | 1.4579 0.1070 10 0.4541 |
Kidneys (g) | 91 [a] | Mean SD N %Diff | 3.0645 0.2430 10 - | 2.7847 0.3149 10 -9.1316 | 2.9595 0.3123 10 -3.4286 | 3.0083 0.2526 10 -1.8332 |
Liver (g) | 91 [a] | Mean SD N %Diff | 13.2163 1.3875 10 - | 12.4174 1.6967 10 -6.0453 | 12.9404 1.1575 10 -2.0878 | 13.0536 1.7073 10 -1.2309 |
[a] - Anova & Dunnett
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Pituitary (g) | 103b [a] | Mean SD N %Diff | 0.0152 0.0014 10 - | 0.0149 0.0026 10 -2.3653 | 0.0147 0.0012 10 -3.2194 | 0.0146 0.0013 10 -4.1393 |
Prostate (g) | 91 [a1] | Mean SD N %Diff | 1.3174 0.2153 10 . | 1.3204 0.2125 10 0.2277 | 1.3592 0.1831 10 3.1698 | 1.3574 0.2370 10 3.0355 |
Seminal vesicles & coagulating glands (g) | 91 [a1] | Mean SD N %Diff | 1.8548 0.3517 10 - | 1.8582 0.2892 10 0.1865 | 1.7220 0.2682 10 -7.1599 | 1.6366 0.2224 10 -11.7642 |
Spleen (g) | 91 [a] | Mean SD N %Diff | 1.1633 0.3505 10 - | 0.9371 0.2472 10 -19.4385 | 0.9102 0.0729 10 -21.7518 | 0.9859 0.1282 10 -15.2502 |
Testes (g) | 91 [a] | Mean SD N %Diff | 3.9520 0.2849 10 - | 4.0709 0.3147 10 3.0073 | 4.4664 0.9630 10 13.0143 | 4.1772 0.9146 10 5.6971 |
Thymus (g) | 91 [a1] | Mean SD N %Diff | 0.4240 0.0807 10 . | 0.4419 0.0954 10 4.2266 | 0.4314 0.0884 10 1.7595 | 0.4439 0.0984 10 4.7054 |
Thyroid with parathyroids (g) | 103b [a1] | Mean SD N %Diff | 0.0345 0.0039 10 - | 0.0348 0.0075 10 0.9576 | 0.0366 0.0053 10 6.2101 | 0.0367 0.0069 10 6.4423 |
[a] - Anova & Dunnett(Rank); b: Weighed after fixation
[a1] Anova & Dunnett
Appendix 15, TABLE 18. Summary of Terminal Fasting Body Weights and Organ to Body Weight Ratios on Day 91 – Males
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Terminal Fasting BW (g) | 91 [a] | Mean SD N %Diff | 509.88 49.34 10 - | 478.23 55.04 10 -6.21 | 491.21 37.99 10 -3.66 | 495.90 38.71 10 -2.74 |
Adrenals (%) | 91 [a1] | Mean SD N %Diff | 0.0150 0.0021 10 . | 0.0144 0.0020 10 -3.7578 | 0.0141 0.0018 10 -5.5958 | 0.0150 0.0032 10 0.0158 |
Brain (%) | 91 [a] | Mean SD N %Diff | 0.4319 0.0375 10 . | 0.4599 0.0422 10 6.4924 | 0.4541 0.0380 10 5.1272 | 0.4420 0.0286 10 2.3426 |
Epididymides (%) | 91 [a] | Mean SD N %Diff | 0.3226 0.0287 10 . | 0.3354 0.0337 10 3.9506 | 0.3148 0.0294 10 -2.4277 | 0.3298 0.0490 10 2.2104 |
Heart (%) | 91 [a] | Mean SD N %Diff | 0.2859 0.0233 10 . | 0.2982 0.0215 10 4.2923 | 0.3018 0.0219 10 5.5438 | 0.2945 0.0163 10 2.9834 |
Kidneys (%) | 91 [a] | Mean SD N %Diff | 0.6037 0.0507 10 . | 0.5837 0.0463 10 -3.3054 | 0.6028 0.0479 10 -0.1498 | 0.6072 0.0348 10 0.5810 |
Liver (%) | 91 [a] | Mean SD N %Diff | 2.5931 0.1411 10 . | 2.5957 0.1589 10 0.0993 | 2.6367 0.1752 10 1.6800 | 2.6237 0.1589 10 1.1796 |
[a] - Anova & Dunnett
[a1] Anova & Dunnett(Log)
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Pituitary (%) | 103b [a] | Mean SD N %Diff | 0.0030 0.0004 10 . | 0.0031 0.0006 10 4.4058 | 0.0030 0.0003 10 0.2903 | 0.0029 0.0002 10 -2.0435 |
Prostate (%) | 91 [a1] | Mean SD N %Diff | 0.2618 0.0565 10 . | 0.2770 0.0415 10 5.8307 | 0.2779 0.0394 10 6.1614 | 0.2763 0.0600 10 5.5593 |
Seminal vesicles & coagulating glands (%) | 91 [a2] | Mean SD N %Diff | 0.3657 0.0686 10 . | 0.3935 0.0800 10 7.5991 | 0.3535 0.0716 10 -3.3203 | 0.3319 0.0507 10 -9.2462 |
Spleen (%) | 91 [a] | Mean SD N %Diff | 0.2294 0.0722 10 . | 0.1983 0.0579 10 -13.5662 | 0.1863 0.0212 10 -18.7774 | 0.1999 0.0289 10 -12.8569 |
Testes (%) | 91 [a] | Mean SD N %Diff | 0.7815 0.0947 10 . | 0.8577 0.0818 10 9.7452 | 0.9055 * 0.1463 10 15.8598 | 0.8503 0.2251 10 8.7921 |
Thymus (%) | 91 [a2] | Mean SD N %Diff | 0.0828 0.0112 10 . | 0.0938 0.0252 10 13.2315 | 0.0881 0.0185 10 6.4465 | 0.0897 0.0204 10 8.3523 |
Thyroid with parathyroids (%) | 103b [a] | Mean SD N %Diff | 0.0068 0.0007 10 . | 0.0074 0.0019 10 9.0421 | 0.0075 0.0013 10 10.7394 | 0.0074 0.0013 10 9.2319 |
[a] - Anova & Dunnett(Rank): *: Significantly different from vehicle control group at p < 0.05 [a1] - Anova & Dunnett; b: Weighed after fixation
[a2] Anova & Dunnett(Log)
Appendix 15, TABLE 19. Summary of Brain Weights and Organ to Brain Weight Ratios on Day 91 – Males
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Brain (g) | 91 [a] | Mean SD N %Diff | 2.1871 0.0848 10 - | 2.1804 0.0952 10 -0.3050 | 2.2186 0.0756 10 1.4398 | 2.1833 0.0823 10 -0.1747 |
Adrenals (%) | 91 [a] | Mean SD N %Diff | 3.4790 0.5169 10 . | 3.1474 0.4738 10 -9.5312 | 3.1298 0.4766 10 -10.0379 | 3.3800 0.6560 10 -2.8465 |
Epididymides (%) | 91 [a] | Mean SD N %Diff | 74.9358 6.4603 10 . | 73.0190 5.0640 10 -2.5580 | 69.5946 7.2156 10 -7.1278 | 74.4686 9.0070 10 -0.6235 |
Heart (%) | 91 [a] | Mean SD N %Diff | 66.3512 4.2154 10 . | 65.1155 4.9748 10 -1.8623 | 66.8264 7.0001 10 0.7162 | 66.7903 4.4849 10 0.6618 |
Kidneys (%) | 91 [a] | Mean SD N %Diff | 140.3386 12.7030 10 . | 127.5292 11.3593 10 -9.1275 | 133.3486 12.6854 10 -4.9808 | 137.7556 10.0786 10 -1.8406 |
Liver (%) | 91 [a] | Mean SD N %Diff | 603.5938 50.4892 10 . | 568.8458 65.9076 10 -5.7568 | 583.9523 57.7208 10 -3.2541 | 597.6385 73.0185 10 -0.9866 |
Pituitary (%) | 103b [a1] | Mean SD N %Diff | 0.6965 0.0651 10 . | 0.6809 0.1094 10 -2.2338 | 0.6647 0.0594 10 -4.5662 | 0.6680 0.0527 10 -4.0882 |
[a] - Anova & Dunnett; b: Weighed after fixation
[a1] Anova & Dunnett(Rank)
Sex: Male Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Prostate (%) | 91 [a] | Mean SD N %Diff | 60.1040 8.8678 10 . | 60.5553 9.3413 10 0.7508 | 61.4339 9.3932 10 2.2126 | 62.2769 11.3167 10 3.6152 |
Seminal vesicles & coagulating glands (%) | 91 [a] | Mean SD N %Diff | 84.5021 13.9828 10 . | 85.6124 15.5667 10 1.3139 | 77.5066 10.8812 10 -8.2785 | 74.9204 9.3218 10 -11.3390 |
Spleen (%) | 91 [a1] | Mean SD N %Diff | 53.3096 16.7113 10 . | 42.8106 10.7724 10 -19.6943 | 41.0098 * 2.6829 10 -23.0725 | 45.1351 5.2933 10 -15.3340 |
Testes (%) | 91 [a1] | Mean SD N %Diff | 180.6955 10.7228 10 . | 186.7867 12.9311 10 3.3710 | 201.3298 42.7738 10 11.4194 | 191.5330 42.5190 10 5.9977 |
Thymus (%) | 91 [a] | Mean SD N %Diff | 19.4011 3.7557 10 . | 20.2604 4.3009 10 4.4290 | 19.5627 4.6970 10 0.8330 | 20.3386 4.5300 10 4.8318 |
Thyroid with parathyroids (%) | 103b [a1] | Mean SD N %Diff | 1.5764 0.1795 10 . | 1.5965 0.3402 10 1.2736 | 1.6487 0.2197 10 4.5865 | 1.6786 0.3001 10 6.4834 |
[a] - Anova & Dunnett; b: Weighed after fixation
[a1] Anova & Dunnett(Rank): *: Significantly different from vehicle control group at p < 0.05
Appendix 15, TABLE 20. Summary of Terminal Fasting Body Weights and Organ Weights on Day 119 – Males
Sex: Male Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Terminal Fasting BW (g) | Day 119 [p] | Mean SD N %Diff | 521.68 40.03 5 - | 560.05 63.66 5 7.36 |
Adrenals (g) | Day 119 [p] | Mean SD N %Diff | 0.0682 0.0126 5 - | 0.0775 0.0145 5 13.6324 |
Brain (g) | Day 119 [p] | Mean SD N %Diff | 2.1785 0.0994 5 - | 2.2669 0.0962 5 4.0606 |
Epididymides (g) | Day 119 [p] | Mean SD N %Diff | 1.6050 0.1204 5 - | 1.7387 0.1539 5 8.3341 |
Heart (g) | Day 119 [p] | Mean SD N %Diff | 1.5181 0.1572 5 - | 1.6032 0.1118 5 5.6029 |
Kidneys (g) | Day 119 [p] | Mean SD N %Diff | 3.0056 0.4252 5 - | 3.3120 0.1087 5 10.1930 |
Liver (g) | Day 119 [p] | Mean SD N %Diff | 14.0219 1.2716 5 - | 14.1928 1.3253 5 1.2191 |
[p] - T-Test: [G1RvsG4R]
Sex: Male Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Pituitary (g) | Day 124b [p] | Mean SD N %Diff | 0.0145 0.0012 5 - | 0.0156 0.0006 5 7.4278 |
Prostate (g) | Day 119 [p] | Mean SD N %Diff | 1.2863 0.2097 5 . | 1.2834 0.1934 5 -0.2270 |
Seminal vesicles & coagulating glands (g) | Day 119 [p] | Mean SD N %Diff | 2.1859 0.4301 5 - | 2.5391 0.2215 5 16.1541 |
Spleen (g) | Day 119 [p] | Mean SD N %Diff | 0.9333 0.0951 5 - | 0.9920 0.1330 5 6.2941 |
Testes (g) | Day 119 [p] | Mean SD N %Diff | 4.0280 0.6371 5 - | 4.4491 0.8062 5 10.4532 |
Thymus (g) | Day 119 [p] | Mean SD N %Diff | 0.3881 0.0831 5 . | 0.3699 0.1002 5 -4.7047 |
Thyroid with parathyroids (g) | Day 124b [p] | Mean SD N %Diff | 0.0296 0.0023 5 - | 0.0311 0.0028 5 4.9966 |
[p] - T-Test: [G1RvsG4R] ; b: Weighed after fixation
Appendix 15, TABLE 21. Summary of Terminal Fasting Body Weights and Organ to Body Weight Ratios on Day 119 – Males
Sex: Male Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Terminal Fasting BW (g) | Day 119 [p] | Mean SD N %Diff | 521.68 40.03 5 - | 560.05 63.66 5 7.36 |
Adrenals (%) | Day 119 [p] | Mean SD N %Diff | 0.0131 0.0021 5 . | 0.0140 0.0030 5 6.7155 |
Brain (%) | Day 119 [p] | Mean SD N %Diff | 0.4186 0.0203 5 . | 0.4082 0.0406 5 -2.4909 |
Epididymides (%) | Day 119 [p] | Mean SD N %Diff | 0.3093 0.0352 5 . | 0.3138 0.0465 5 1.4529 |
Heart (%) | Day 119 [p] | Mean SD N %Diff | 0.2911 0.0217 5 . | 0.2878 0.0223 5 -1.1221 |
Kidneys (%) | Day 119 [p] | Mean SD N %Diff | 0.5747 0.0539 5 . | 0.5963 0.0585 5 3.7572 |
Liver (%) | Day 119 [p] | Mean SD N %Diff | 2.6866 0.0817 5 . | 2.5395 * 0.0847 5 -5.4741 |
[p] - T-Test: [G1RvsG4R]
{Liver.} G1RvsG4R: *: Significantly different from vehicle control group at p < 0.05
Sex: Male Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Pituitary (%) | Day 124b [p] | Mean SD N %Diff | 0.0028 0.0003 5 . | 0.0028 0.0003 5 0.5069 |
Prostate (%) | Day 119 [p] | Mean SD N %Diff | 0.2465 0.0364 5 . | 0.2288 0.0175 5 -7.1964 |
Seminal vesicles & coagulating glands (%) | Day 119 [p] | Mean SD N %Diff | 0.4205 0.0854 5 . | 0.4575 0.0632 5 8.7997 |
Spleen (%) | Day 119 [p] | Mean SD N %Diff | 0.1789 0.0134 5 . | 0.1774 0.0179 5 -0.8544 |
Testes (%) | Day 119 [p] | Mean SD N %Diff | 0.7729 0.1173 5 . | 0.8072 0.2077 5 4.4404 |
Thymus (%) | Day 119 [p] | Mean SD N %Diff | 0.0746 0.0158 5 . | 0.0656 0.0150 5 -12.0535 |
Thyroid with parathyroids (%) | Day 124b [p1] | Mean SD N %Diff | 0.0057 0.0004 5 . | 0.0056 0.0003 5 -2.0224 |
[p] - T-Test: [G1RvsG4R] ; b: Weighed after fixation
[p1] - Wilcoxon(Rank): [G1RvsG4R]
Appendix 15, TABLE 22. Summary of Brain Weights and Organ to Brain Weight Ratios on Day 119 – Males
Sex: Male Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Brain (g) | Day 119 [p] | Mean SD N %Diff | 2.1785 0.0994 5 - | 2.2669 0.0962 5 4.0606 |
Adrenals (%) | Day 119 [p] | Mean SD N %Diff | 3.1328 0.5596 5 . | 3.4341 0.6901 5 9.6183 |
Epididymides (%) | Day 119 [p] | Mean SD N %Diff | 73.7359 5.4465 5 . | 76.8589 8.1876 5 4.2354 |
Heart (%) | Day 119 [p] | Mean SD N %Diff | 69.5539 4.2952 5 . | 70.8565 6.1340 5 1.8729 |
Kidneys (%) | Day 119 [p] | Mean SD N %Diff | 137.7725 16.7897 5 . | 146.4011 9.6973 5 6.2629 |
Liver (%) | Day 119 [p] | Mean SD N %Diff | 643.1001 39.6597 5 . | 626.5726 58.1752 5 -2.5700 |
Pituitary (%) | Day 124b [p] | Mean SD N %Diff | 0.6681 0.0536 5 . | 0.6895 0.0251 5 3.2152 |
[p] - T-Test: [G1RvsG4R] ; b: Weighed after fixation
Sex: Male Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Prostate (%) | Day 119 [p] | Mean SD N %Diff | 58.8403 7.4497 5 . | 56.6770 8.6437 5 -3.6766 |
Seminal vesicles & coagulating glands (%) | Day 119 [p] | Mean SD N %Diff | 100.0448 16.7022 5 . | 112.4500 14.0003 5 12.3997 |
Spleen (%) | Day 119 [p] | Mean SD N %Diff | 42.7643 2.7556 5 . | 43.7411 5.3876 5 2.2841 |
Testes (%) | Day 119 [p] | Mean SD N %Diff | 184.3709 23.1031 5 . | 197.3571 42.8986 5 7.0435 |
Thymus (%) | Day 119 [p] | Mean SD N %Diff | 17.8647 3.9596 5 . | 16.2224 3.9026 5 -9.1929 |
Thyroid with parathyroids (%) | Day 124b [p] | Mean SD N %Diff | 1.3584 0.0555 5 . | 1.3733 0.1256 5 1.0992 |
[p] - T-Test: [G1RvsG4R] ; b: Weighed after fixation
Appendix 15, TABLE 23. Summary of Terminal Fasting Body Weights and Organ Weights on Day 91 – Females
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Terminal Fasting BW (g) | 91 [a] | Mean SD N %Diff | 275.98 22.96 10 - | 283.90 22.39 10 2.87 | 274.09 16.62 10 -0.68 | 288.64 27.41 10 4.59 |
Adrenals (g) | 91 [a1] | Mean SD N %Diff | 0.0887 0.0083 10 - | 0.0895 0.0161 10 0.9129 | 0.0933 0.0061 10 5.1505 | 0.0942 0.0102 10 6.1760 |
Brain (g) | 91 [a] | Mean SD N %Diff | 2.0429 0.0704 10 - | 2.0222 0.0982 10 -1.0123 | 2.0281 0.0700 10 -0.7274 | 2.0674 0.0795 10 1.2002 |
Heart (g) | 91 [a1] | Mean SD N %Diff | 0.9845 0.0374 10 - | 1.0288 0.0660 10 4.4945 | 1.0019 0.0798 10 1.7602 | 1.0110 0.0932 10 2.6886 |
Kidneys (g) | 91 [a] | Mean SD N %Diff | 1.7205 0.1478 10 - | 1.7287 0.1405 10 0.4731 | 1.6658 0.1587 10 -3.1839 | 1.7996 0.1636 10 4.5945 |
Liver (g) | 91 [a] | Mean SD N %Diff | 7.5040 0.7841 10 - | 7.9984 0.9514 10 6.5882 | 7.3846 0.5394 10 -1.5917 | 7.6663 0.4731 10 2.1634 |
Ovaries (g) | 91 [a] | Mean SD N %Diff | 0.1118 0.0141 10 - | 0.1269 0.0144 10 13.5009 | 0.1122 0.0181 10 0.3937 | 0.1113 0.0177 10 -0.4295 |
[a] - Anova & Dunnett
[a1] Anova & Dunnett(Rank)
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Pituitary (g) | 103b [a] | Mean SD N %Diff | 0.0177 0.0025 10 - | 0.0185 0.0030 10 4.2301 | 0.0173 0.0040 10 -2.6509 | 0.0173 0.0029 10 -2.4817 |
Spleen (g) | 91 [a] | Mean SD N %Diff | 0.6368 0.0974 10 - | 0.6698 0.0656 10 5.1824 | 0.6868 0.0836 10 7.8631 | 0.6569 0.0907 10 3.1660 |
Thymus (g) | 91 [a] | Mean SD N %Diff | 0.3346 0.0471 10 . | 0.3354 0.0702 10 0.2660 | 0.3772 0.0606 10 12.7485 | 0.3455 0.0612 10 3.2611 |
Thyroid with parathyroids (g) | 103b [a1] | Mean SD N %Diff | 0.0298 0.0063 10 - | 0.0296 0.0036 10 -0.5373 | 0.0306 0.0057 10 2.8543 | 0.0270 0.0031 10 -9.3015 |
Uterus with cervix (g) | 91 [a2] | Mean SD N %Diff | 0.7268 0.1673 10 . | 0.6877 0.0893 10 -5.3704 | 0.9847 0.4090 10 35.4973 | 0.7409 0.2852 10 1.9497 |
Anova & Dunnett;
Anova & Dunnett(Log)
Anova & Dunnett(Rank)
Appendix 15, TABLE 24. Summary of Terminal Fasting Body Weights and Organ to Body Weight Ratios on Day 91 – Females
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Terminal Fasting BW (g) | 91 [a] | Mean SD N %Diff | 275.98 22.96 10 - | 283.90 22.39 10 2.87 | 274.09 16.62 10 -0.68 | 288.64 27.41 10 4.59 |
Adrenals (%) | 91 [a1] | Mean SD N %Diff | 0.0322 0.0030 10 . | 0.0316 0.0054 10 -2.0225 | 0.0342 0.0033 10 5.9907 | 0.0329 0.0043 10 1.9291 |
Brain (%) | 91 [a] | Mean SD N %Diff | 0.7440 0.0541 10 . | 0.7158 0.0589 10 -3.7890 | 0.7424 0.0519 10 -0.2155 | 0.7224 0.0775 10 -2.8947 |
Heart (%) | 91 [a] | Mean SD N %Diff | 0.3593 0.0364 10 . | 0.3639 0.0304 10 1.2928 | 0.3659 0.0268 10 1.8429 | 0.3518 0.0349 10 -2.0858 |
Kidneys (%) | 91 [a2] | Mean SD N %Diff | 0.6251 0.0503 10 . | 0.6098 0.0355 10 -2.4416 | 0.6099 0.0717 10 -2.4368 | 0.6256 0.0541 10 0.0835 |
Liver (%) | 91 [a] | Mean SD N %Diff | 2.7205 0.1903 10 . | 2.8136 0.1908 10 3.4192 | 2.6963 0.1623 10 -0.8929 | 2.6766 0.2985 10 -1.6168 |
Ovaries (%) | 91 [a] | Mean SD N %Diff | 0.0406 0.0048 10 . | 0.0448 0.0050 10 10.3760 | 0.0409 0.0055 10 0.6560 | 0.0390 0.0078 10 -4.0506 |
Anova & Dunnett Anova & Dunnett(Rank)
Anova & Dunnett(Log)
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Pituitary (%) | 103b [a] | Mean SD N %Diff | 0.0065 0.0010 10 . | 0.0066 0.0012 10 1.4967 | 0.0063 0.0015 10 -2.2692 | 0.0060 0.0010 10 -6.7863 |
Spleen (%) | 91 [a] | Mean SD N %Diff | 0.2309 0.0303 10 . | 0.2368 0.0249 10 2.5798 | 0.2504 0.0246 10 8.4454 | 0.2293 0.0365 10 -0.6766 |
Thymus (%) | 91 [a1] | Mean SD N %Diff | 0.1210 0.0110 10 . | 0.1186 0.0258 10 -2.0196 | 0.1383 0.0247 10 14.2483 | 0.1201 0.0221 10 -0.8066 |
Thyroid with parathyroids (%) | 103b [a] | Mean SD N %Diff | 0.0108 0.0021 10 . | 0.0105 0.0013 10 -2.9900 | 0.0112 0.0022 10 3.9089 | 0.0094 0.0014 10 -12.6217 |
Uterus with cervix (%) | 91 [a2] | Mean SD N %Diff | 0.2639 0.0630 10 . | 0.2439 0.0379 10 -7.6065 | 0.3615 0.1556 10 36.9683 | 0.2584 0.1003 10 -2.0991 |
Anova & Dunnett;
Anova & Dunnett(Log)
Anova & Dunnett(Rank)
Appendix 15, TABLE 25. Summary of Brain Weights and Organ to Brain Weight Ratios on Day 91 – Females
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Brain (g) | 91 [a] | Mean SD N %Diff | 2.0429 0.0704 10 - | 2.0222 0.0982 10 -1.0123 | 2.0281 0.0700 10 -0.7274 | 2.0674 0.0795 10 1.2002 |
Adrenals (%) | 91 [a1] | Mean SD N %Diff | 4.3415 0.3593 10 . | 4.4316 0.8093 10 2.0750 | 4.6008 0.2698 10 5.9727 | 4.5661 0.5515 10 5.1736 |
Heart (%) | 91 [a] | Mean SD N %Diff | 48.2463 2.5304 10 . | 51.0142 4.5978 10 5.7370 | 49.4542 4.2601 10 2.5037 | 48.8776 3.7949 10 1.3085 |
Kidneys (%) | 91 [a] | Mean SD N %Diff | 84.2497 7.0691 10 . | 85.6302 7.8237 10 1.6386 | 82.1495 7.3198 10 -2.4928 | 87.0850 7.7178 10 3.3654 |
Liver (%) | 91 [a] | Mean SD N %Diff | 367.3662 36.9055 10 . | 395.4801 42.1633 10 7.6528 | 364.7693 32.3434 10 -0.7069 | 371.0654 22.4820 10 1.0070 |
Ovaries (%) | 91 [a] | Mean SD N %Diff | 5.4819 0.7512 10 . | 6.2745 0.6765 10 14.4589 | 5.5378 0.8738 10 1.0205 | 5.3669 0.7279 10 -2.0970 |
Pituitary (%) | 103b [a] | Mean SD N %Diff | 0.8686 0.1242 10 . | 0.9153 0.1566 10 5.3783 | 0.8523 0.1979 10 -1.8750 | 0.8362 0.1354 10 -3.7309 |
[a] - Anova & Dunnett; b: Weighed after fixation
[a1] Anova & Dunnett(Rank)
Sex: Female Day(s) Relative to Start Date | G1 0 mg/kg/day | G2 111 mg/kg/day | G3 333 mg/kg/day | G4 1000 mg/kg/day | ||
Spleen (%) | 91 [a] | Mean SD N %Diff | 31.1921 4.8330 10 . | 33.1079 2.6272 10 6.1419 | 33.9355 4.4713 10 8.7951 | 31.7195 3.6721 10 1.6908 |
Thymus (%) | 91 [a1] | Mean SD N %Diff | 16.3847 2.2777 10 . | 16.5824 3.4157 10 1.2068 | 18.6606 3.3939 10 13.8905 | 16.6820 2.6937 10 1.8145 |
Thyroid with parathyroids (%) | 103b [a] | Mean SD N %Diff | 1.4555 0.2912 10 . | 1.4653 0.1703 10 0.6801 | 1.5131 0.2947 10 3.9596 | 1.3079 0.1598 10 -10.1370 |
Uterus with cervix (%) | 91 [a2] | Mean SD N %Diff | 35.5778 8.2525 10 . | 33.9622 3.5255 10 -4.5412 | 49.0565 21.7419 10 37.8850 | 35.6692 12.8037 10 0.2569 |
[a] - Anova & Dunnett; [a1] - Anova & Dunnett(Log)
[a2] Anova & Dunnett(Rank)
Appendix 15, TABLE 26. Summary of Terminal Fasting Body Weights and Organ Weights on Day 119 – Females
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Terminal Fasting BW (g) | Day 119 [p] | Mean SD N %Diff | 293.36 16.57 5 - | 308.97 25.53 5 5.32 |
Adrenals (g) | Day 119 [p1] | Mean SD N %Diff | 0.0917 0.0158 5 - | 0.1034 0.0173 5 12.8300 |
Brain (g) | Day 119 [p2] | Mean SD N %Diff | 2.0873 0.0470 5 - | 2.1181 0.1572 5 1.4756 |
Heart (g) | Day 119 [p2] | Mean SD N %Diff | 1.0674 0.1027 5 - | 1.0941 0.1462 5 2.4976 |
Kidneys (g) | Day 119 [p2] | Mean SD N %Diff | 1.8137 0.2827 5 - | 1.9450 0.2069 5 7.2439 |
Liver (g) | Day 119 [p] | Mean SD N %Diff | 8.3472 1.1153 5 - | 8.3153 1.2175 5 -0.3829 |
Ovaries (g) | Day 119 [p] | Mean SD N %Diff | 0.1200 0.0160 5 - | 0.0875 * 0.0257 5 -27.1121 |
[p] - Wilcoxon(Rank): [G1RvsG4R]
{Ovaries} G1RvsG4R: *: Significantly different from vehicle control group at p < 0.05
[p1] - T-Test(Log): [G1RvsG4R]
[p2] T-Test: [G1RvsG4R]
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Pituitary (g) | Day 124b [p] | Mean SD N %Diff | 0.0177 0.0025 5 - | 0.0188 0.0044 5 6.2288 |
Spleen (g) | Day 119 [p] | Mean SD N %Diff | 0.6260 0.1032 5 - | 0.6587 0.1059 5 5.2169 |
Thymus (g) | Day 119 [p1] | Mean SD N %Diff | 0.3458 0.0447 5 . | 0.3144 0.0458 5 -9.0967 |
Thyroid with parathyroids (g) | Day 124b [p1] | Mean SD N %Diff | 0.0276 0.0019 5 - | 0.0278 0.0042 5 0.5069 |
Uterus with cervix (g) | Day 119 [p1] | Mean SD N %Diff | 0.9721 0.2480 5 . | 1.0763 0.1814 5 10.7255 |
[p] - Wilcoxon(Rank): [G1RvsG4R] ; b: Weighed after fixation
[p1] T-Test: [G1RvsG4R]
Appendix 15, TABLE 27. Summary of Terminal Fasting Body Weights and Organ to Body Weight Ratios on Day 119 – Females
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Terminal Fasting BW (g) | Day 119 [p] | Mean SD N %Diff | 293.36 16.57 5 - | 308.97 25.53 5 5.32 |
Adrenals (%) | Day 119 [p1] | Mean SD N %Diff | 0.0311 0.0036 5 . | 0.0333 0.0030 5 7.1958 |
Brain (%) | Day 119 [p1] | Mean SD N %Diff | 0.7131 0.0377 5 . | 0.6887 0.0709 5 -3.4179 |
Heart (%) | Day 119 [p1] | Mean SD N %Diff | 0.3635 0.0208 5 . | 0.3543 0.0403 5 -2.5080 |
Kidneys (%) | Day 119 [p1] | Mean SD N %Diff | 0.6159 0.0670 5 . | 0.6294 0.0398 5 2.1967 |
Liver (%) | Day 119 [p1] | Mean SD N %Diff | 2.8368 0.2202 5 . | 2.6833 0.2081 5 -5.4091 |
Ovaries (%) | Day 119 [p1] | Mean SD N %Diff | 0.0409 0.0053 5 . | 0.0283 * 0.0086 5 -30.7594 |
[p] - Wilcoxon(Rank): [G1RvsG4R]
[p1] - T-Test: [G1RvsG4R]
{Ovaries} G1RvsG4R: *: Significantly different from vehicle control group at p < 0.05
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Pituitary (%) | Day 124b [p] | Mean SD N %Diff | 0.0060 0.0007 5 . | 0.0061 0.0018 5 2.2410 |
Spleen (%) | Day 119 [p] | Mean SD N %Diff | 0.2125 0.0232 5 . | 0.2125 0.0204 5 -0.0015 |
Thymus (%) | Day 119 [p] | Mean SD N %Diff | 0.1178 0.0129 5 . | 0.1020 0.0151 5 -13.4214 |
Thyroid with parathyroids (%) | Day 124b [p] | Mean SD N %Diff | 0.0094 0.0007 5 . | 0.0090 0.0010 5 -4.8501 |
Uterus with cervix (%) | Day 119 [p] | Mean SD N %Diff | 0.3343 0.0992 5 . | 0.3481 0.0506 5 4.1434 |
[p] - T-Test: [G1RvsG4R] ; b: Weighed after fixation
Appendix 15, TABLE 28. Summary of Brain Weights and Organ to Brain Weight Ratios on Day 119 – Females
Sex: FemaleDay(s) Relative to Start Date | G1R0 mg/kg/day | G4R1000 mg/kg/day | ||
Brain (g) | Day 119 [p] | MeanSDN%Diff | 2.08730.04705- | 2.11810.157251.4756 |
Adrenals (%) | Day 119 [p] | MeanSDN%Diff | 4.39250.75595. | 4.90930.9212511.7651 |
Heart (%) | Day 119 [p] | MeanSDN%Diff | 51.11434.45465. | 51.86647.872551.4714 |
Kidneys (%) | Day 119 [p1] | MeanSDN%Diff | 86.834412.97785. | 92.442713.706856.4586 |
Liver (%) | Day 119 [p1] | MeanSDN%Diff | 400.020053.79435. | 393.694359.52925-1.5813 |
Ovaries (%) | Day 119 [p] | MeanSDN%Diff | 5.75140.77655. | 4.0846 * 1.05245-28.9809 |
Pituitary (%) | Day 124b [p2] | MeanSDN%Diff | 0.84570.11255. | 0.89950.281556.3657 |
[p] - T-Test: [G1RvsG4R] ; b: Weighed after fixation
{Ovaries} G1RvsG4R: *: Significantly different from vehicle control group at p < 0.05
[p1] - Wilcoxon(Rank): [G1RvsG4R]
[p2] - T-Test(Log): [G1RvsG4R]
Sex: Female Day(s) Relative to Start Date | G1R 0 mg/kg/day | G4R 1000 mg/kg/day | ||
Spleen (%) | Day 119 [p] | Mean SD N %Diff | 29.9853 4.8725 5 . | 31.2922 5.8740 5 4.3584 |
Thymus (%) | Day 119 [p1] | Mean SD N %Diff | 16.5505 1.9143 5 . | 14.8833 2.1876 5 -10.0731 |
Thyroid with parathyroids (%) | Day 124b [p1] | Mean SD N %Diff | 1.3240 0.0954 5 . | 1.3145 0.2039 5 -0.7193 |
Uterus with cervix (%) | Day 119 [p1] | Mean SD N %Diff | 46.6818 12.6254 5 . | 50.7888 7.4906 5 8.7980 |
[p] - Wilcoxon(Rank): [G1RvsG4R] ; b: Weighed after fixation [p1] - T-Test: [G1RvsG4R]
Appendix 15, TABLE 29. Summary of Sperm Parameters
Group No. Dose (mg/kg/day) |
| Sperm Motility | Sperm Morphology | Cauda Epididymal Sperm Counts | ||||
Percentage of progressive motile sperms | Percentage of motile sperms | Percentage of normal sperms | Percentage of abnormal sperms | Cauda epididymis weight (g) | No. of sperms per cauda epididymis (x106) | No. of sperms per gram of cauda epididymis (x106) | ||
G1 | Mean | 64.40 | 86.40 | 99.05 | 0.95 | 0.32 | 250.78 | 793.73 |
0 | SD | 8.13 | 6.70 | 1.01 | 1.01 | 0.02 | 17.00 | 84.79 |
| n | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
G2 | Mean | 66.00 | 87.70 |
|
|
|
|
|
111 | SD | 6.86 | 5.06 | NA | NA | NA | NA | NA |
| n | 10 | 10 |
|
|
|
|
|
G3 | Mean | 64.60 | 85.20 |
|
|
|
|
|
333 | SD | 10.24 | 7.74 | NA | NA | NA | NA | NA |
| n | 10 | 10 |
|
|
|
|
|
G4 | Mean | 62.60 | 85.10 | 97.25 | 2.75 | 0.30 | 235.30 | 774.63 |
1000 | SD | 8.57 | 6.66 | 2.62 | 2.62 | 0.02 | 26.41 | 51.20 |
| n | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
G1R | Mean | 61.00 | 85.20 |
|
|
|
|
|
0 | SD | 15.36 | 7.98 | NA | NA | NA | NA | NA |
| n | 5 | 5 |
|
|
|
|
|
G4R | Mean | 61.40 | 82.20 |
|
|
|
|
|
1000 | SD | 7.47 | 6.83 | NA | NA | NA | NA | NA |
| n | 5 | 5 |
|
|
|
|
|
Appendix 15, TABLE 30. Summary of Gross Pathology Findings
----------------------- MALES ----------------------- ---------------------- FEMALES ----------------------
Removal Reason: TERMINAL SACRIFICE
Group No.: G1 G2 G3 G4 G1R G4R G1 G2 G3 G4 G1R G4R
Dose (mg/kg/day): 0 111 333 1000 0 1000 0 111 333 1000 0 1000
Number of Animals on Study : 10 10 10 10 5 5 10 10 10 10 5 5 Number of Animals Completed: (10) (10) (10) (10) (5) (5) (10) (10) (10) (10) (5) (5)
CECUM;
Submitted....................................... (10) (10) (10) (10) (5) (5) (10) (10) (10) (10) (5) (5) No Visible Lesions.............................. 10 3 0 0 5 5 10 3 0 0 5 5 Contents orange ................................ 0 7 10 10 0 0 0 7 10 10 0 0
COLON;
Submitted....................................... (10) (10) (10) (10) (5) (5) (10) (10) (10) (10) (5) (5) No Visible Lesions.............................. 10 8 4 6 5 5 10 9 4 4 5 5 Contents orange ................................ 0 2 6 4 0 0 0 1 6 6 0 0
ILEUM;
Submitted....................................... (10) (10) (10) (10) (5) (5) (10) (10) (10) (10) (5) (5) No Visible Lesions.............................. 10 8 9 9 5 5 10 10 9 9 5 5 Contents orange ................................ 0 2 1 1 0 0 0 0 1 1 0 0
RECTUM;
Submitted....................................... (10) (10) (10) (10) (5) (5) (10) (10) (10) (10) (5) (5)
No Visible Lesions.............................. 10 10 6 6 5 5 10 9 5 5 5 5 Contents orange ................................ 0 0 4 4 0 0 0 1 5 5 0 0
TESTES;
Submitted....................................... (10) (10) (10) (10) (5) (5) (-) (-) (-) (-) (-) (-) No Visible Lesions.............................. 10 10 8 9 5 5 - - - - - -
Flaccid; bilateral ............................. 0 0 1 1 0 0 - - - - - -
Flaccid; unilateral ............................ 0 0 1 0 0 0 - - - - - -
----------------------- MALES ----------------------- ---------------------- FEMALES ----------------------
Removal Reason: TERMINAL SACRIFICE
Group No.: G1 G2 G3 G4 G1R G4R G1 G2 G3 G4 G1R G4R
Dose (mg/kg/day): 0 111 333 1000 0 1000 0 111 333 1000 0 1000
Number of Animals on Study : 10 10 10 10 5 5 10 10 10 10 5 5 Number of Animals Completed: (10) (10) (10) (10) (5) (5) (10) (10) (10) (10) (5) (5)
UTERUS;
Submitted....................................... (-) (-) (-) (-) (-) (-) (10) (10) (10) (10) (5) (5) No Visible Lesions.............................. - - - - - - 10 10 8 9 4 3 Dilated ........................................ - - - - - - 0 0 2 1 1 2
Note: All the tissues were examined grossly and no gross abnormalities were observed for all the animals.
Appendix 15, TABLE 31. Summary of Histopathology Findings
Dose (mg/kg/day): 0 111 333 1000 0 1000 0 111 333 1000 0 1000
Number of Animals on Study : 10 10 10 10 5 5 10 10 10 10 5 5 Number of Animals Completed: (10) (10) (10) (10) (5) (5) (10) (10) (10) (10) (5) (5)
GLANDS, ADRENAL;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0) Within Normal Limits............................ 8 0 0 8 0 0 10 0 0 10 0 0 Vacuolation; cortical; bilateral ............... (2) (0) (0) (2) (0) (0) (0) (0) (0) (0) (0) (0) minimal ...................................... 2 0 0 2 0 0 0 0 0 0 0 0
AORTA;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
BONE MARROW SMEAR;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
BRAIN CEREBRUM;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
BRAIN CEREBELLUM;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
BRAIN-MEDULLA OBLONGATA;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
Observations: Non Neo-Plastic ----------------------- MALES ----------------------- ---------------------- FEMALES ----------------------
Removal Reason: TERMINAL SACRIFICE
Group No.: G1 G2 G3 G4 G1R G4R G1 G2 G3 G4 G1R G4R
Dose (mg/kg/day): 0 111 333 1000 0 1000 0 111 333 1000 0 1000 Number of Animals on Study : 10 10 10 10 5 5 10 10 10 10 5 5 Number of Animals Completed: (10) (10) (10) (10) (5) (5) (10) (10) (10) (10) (5) (5)
BRAIN-PONS;
Examined........................................ (9) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0) Within Normal Limits............................ 9 0 0 10 0 0 10 0 0 10 0 0 Not Examined: NOT PRESENT ...................... 1 0 0 0 0 0 0 0 0 0 0 0
CECUM;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
CERVIX;
Examined........................................ (-) (-) (-) (-) (-) (-) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ - - - - - - 10 0 0 10 0 0
COLON;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
COAGULATING GLAND;
Examined........................................ (10) (0) (0) (10) (0) (0) (-) (-) (-) (-) (-) (-)
Within Normal Limits............................ 10 0 0 10 0 0 - - - - - -
DUODENUM;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
Observations: Non Neo-Plastic ----------------------- MALES ----------------------- ---------------------- FEMALES ----------------------
Removal Reason: TERMINAL SACRIFICE
Group No.: G1 G2 G3 G4 G1R G4R G1 G2 G3 G4 G1R G4R
Dose (mg/kg/day): 0 111 333 1000 0 1000 0 111 333 1000 0 1000
Number of Animals on Study : 10 10 10 10 5 5 10 10 10 10 5 5 Number of Animals Completed: (10) (10) (10) (10) (5) (5) (10) (10) (10) (10) (5) (5)
ESOPHAGUS;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
EYES;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
BONE, FEMUR/JOINT, FEMOROTIBIAL;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
FEMORAL MUSCLE;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
GUT ASSOCIATED LYMPHOID TISSUE;
Examined........................................ (10) (0) (0) (9) (0) (0) (10) (0) (0) (10) (0) (0) Within Normal Limits............................ 10 0 0 9 0 0 10 0 0 10 0 0 Not Examined: NOT PRESENT ...................... 0 0 0 1 0 0 0 0 0 0 0 0
HEART;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0) Within Normal Limits............................ 8 0 0 7 0 0 10 0 0 10 0 0 Infiltration; mononuclear cell ................. (2) (0) (0) (3) (0) (0) (0) (0) (0) (0) (0) (0) minimal ...................................... 2 0 0 3 0 0 0 0 0 0 0 0
Observations: Non Neo-Plastic ----------------------- MALES ----------------------- ---------------------- FEMALES ----------------------
Removal Reason: TERMINAL SACRIFICE
Group No.: G1 G2 G3 G4 G1R G4R G1 G2 G3 G4 G1R G4R
Dose (mg/kg/day): 0 111 333 1000 0 1000 0 111 333 1000 0 1000
Number of Animals on Study : 10 10 10 10 5 5 10 10 10 10 5 5 Number of Animals Completed: (10) (10) (10) (10) (5) (5) (10) (10) (10) (10) (5) (5)
ILEUM;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
JEJUNUM;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
KIDNEYS;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0) Within Normal Limits............................ 10 0 0 9 0 0 8 0 0 9 0 0 Mineralization; corticomedullary junction;
unilateral ............................... (0) (0) (0) (0) (0) (0) (2) (0) (0) (1) (0) (0) minimal ...................................... 0 0 0 0 0 0 2 0 0 1 0 0 Casts; tubular; bilateral ...................... (0) (0) (0) (1) (0) (0) (0) (0) (0) (0) (0) (0) minimal ...................................... 0 0 0 1 0 0 0 0 0 0 0 0
LARYNX;
Examined........................................ (10) (0) (0) (9) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 9 0 0 10 0 0 10 0 0
Not Examined: NOT PRESENT ...................... 0 0 0 1 0 0 0 0 0 0 0 0
Observations: Non Neo-Plastic ----------------------- MALES ----------------------- ---------------------- FEMALES ----------------------
Removal Reason: TERMINAL SACRIFICE
Group No.: G1 G2 G3 G4 G1R G4R G1 G2 G3 G4 G1R G4R
Dose (mg/kg/day): 0 111 333 1000 0 1000 0 111 333 1000 0 1000
Number of Animals on Study : 10 10 10 10 5 5 10 10 10 10 5 5 Number of Animals Completed: (10) (10) (10) (10) (5) (5) (10) (10) (10) (10) (5) (5)
LEFT EPIDIDYMIS;
Examined........................................ (10) (0) (2) (10) (0) (0) (-) (-) (-) (-) (-) (-) Within Normal Limits............................ 10 0 1 9 0 0 - - - - - - Reduced sperm; luminal ......................... (0) (0) (1) (0) (0) (0) (-) (-) (-) (-) (-) (-) marked ....................................... 0 0 1 0 0 0 - - - - - - Cell debris; luminal ........................... (0) (0) (1) (1) (0) (0) (-) (-) (-) (-) (-) (-) minimal ...................................... 0 0 1 1 0 0 - - - - - -
LIVER;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 4 0 0 2 0 0 10 0 0 9 0 0 Vacuolation; hepatocyte ........................ (6) (0) (0) (7) (0) (0) (0) (0) (0) (1) (0) (0) minimal ...................................... 4 0 0 6 0 0 0 0 0 1 0 0 mild ......................................... 2 0 0 1 0 0 0 0 0 0 0 0 Infiltration; mononuclear cell ................. (0) (0) (0) (1) (0) (0) (0) (0) (0) (0) (0) (0) minimal ...................................... 0 0 0 1 0 0 0 0 0 0 0 0
LUNGS (WITH BRONCHI AND BRONCHIOLES);
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0) Within Normal Limits............................ 9 0 0 8 0 0 10 0 0 9 0 0 Mineralization; blood vessels .................. (1) (0) (0) (2) (0) (0) (0) (0) (0) (1) (0) (0) minimal ...................................... 1 0 0 2 0 0 0 0 0 1 0 0
MAMMARY GLANDS;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
Observations: Non Neo-Plastic ----------------------- MALES ----------------------- ---------------------- FEMALES ----------------------
Removal Reason: TERMINAL SACRIFICE
Group No.: G1 G2 G3 G4 G1R G4R G1 G2 G3 G4 G1R G4R
Dose (mg/kg/day): 0 111 333 1000 0 1000 0 111 333 1000 0 1000
Number of Animals on Study : 10 10 10 10 5 5 10 10 10 10 5 5 Number of Animals Completed: (10) (10) (10) (10) (5) (5) (10) (10) (10) (10) (5) (5)
LYMPH NODES, MESENTERIC;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
LYMPH NODES, MANDIBULAR;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
NERVE, OPTIC;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
OVARIES;
Examined........................................ (-) (-) (-) (-) (-) (-) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ - - - - - - 10 0 0 10 0 0
OVIDUCTS;
Examined........................................ (-) (-) (-) (-) (-) (-) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ - - - - - - 10 0 0 10 0 0
PANCREAS;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
PARATHYROID GLAND;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
Observations: Non Neo-Plastic ----------------------- MALES ----------------------- ---------------------- FEMALES ----------------------
Removal Reason: TERMINAL SACRIFICE
Group No.: G1 G2 G3 G4 G1R G4R G1 G2 G3 G4 G1R G4R
Dose (mg/kg/day): 0 111 333 1000 0 1000 0 111 333 1000 0 1000
Number of Animals on Study : 10 10 10 10 5 5 10 10 10 10 5 5 Number of Animals Completed: (10) (10) (10) (10) (5) (5) (10) (10) (10) (10) (5) (5)
PHARYNX;
Examined........................................ (10) (0) (0) (9) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 9 0 0 10 0 0 10 0 0
Not Examined: NOT PRESENT ...................... 0 0 0 1 0 0 0 0 0 0 0 0
PITUITARY GLAND;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0) Within Normal Limits............................ 8 0 0 9 0 0 10 0 0 10 0 0 Cyst; pars distalis ............................ (2) (0) (0) (1) (0) (0) (0) (0) (0) (0) (0) (0) present ...................................... 2 0 0 1 0 0 0 0 0 0 0 0
PROSTATE;
Examined........................................ (10) (0) (0) (10) (0) (0) (-) (-) (-) (-) (-) (-) Within Normal Limits............................ 8 0 0 9 0 0 - - - - - - Infiltration; mononuclear cell ................. (2) (0) (0) (1) (0) (0) (-) (-) (-) (-) (-) (-) minimal ...................................... 2 0 0 1 0 0 - - - - - -
RECTUM;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
GLANDS, SALIVARY;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0) Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
Observations: Non Neo-Plastic ----------------------- MALES ----------------------- ---------------------- FEMALES ----------------------
Removal Reason: TERMINAL SACRIFICE
Group No.: G1 G2 G3 G4 G1R G4R G1 G2 G3 G4 G1R G4R
Dose (mg/kg/day): 0 111 333 1000 0 1000 0 111 333 1000 0 1000
Number of Animals on Study : 10 10 10 10 5 5 10 10 10 10 5 5 Number of Animals Completed: (10) (10) (10) (10) (5) (5) (10) (10) (10) (10) (5) (5)
NERVE, SCIATIC;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
SEMINAL VESICLE;
Examined........................................ (10) (0) (0) (10) (0) (0) (-) (-) (-) (-) (-) (-)
Within Normal Limits............................ 10 0 0 10 0 0 - - - - - -
SKIN;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
SPINAL CORD;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
SPLEEN;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0) Within Normal Limits............................ 9 0 0 8 0 0 9 0 0 9 0 0 Extramedullary hematopoiesis; increased ........ (1) (0) (0) (2) (0) (0) (1) (0) (0) (1) (0) (0) minimal ...................................... 1 0 0 2 0 0 1 0 0 1 0 0
STERNUM WITH MARROW;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
Observations: Non Neo-Plastic ----------------------- MALES ----------------------- ---------------------- FEMALES ----------------------
Removal Reason: TERMINAL SACRIFICE
Group No.: G1 G2 G3 G4 G1R G4R G1 G2 G3 G4 G1R G4R
Dose (mg/kg/day): 0 111 333 1000 0 1000 0 111 333 1000 0 1000
Number of Animals on Study : 10 10 10 10 5 5 10 10 10 10 5 5 Number of Animals Completed: (10) (10) (10) (10) (5) (5) (10) (10) (10) (10) (5) (5)
STOMACH;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
TESTES;
Examined........................................ (10) (0) (2) (10) (0) (0) (-) (-) (-) (-) (-) (-)
Within Normal Limits............................ 0 0 0 0 0 0 - - - - - - Atrophy; seminiferous tubules; unilateral ...... (0) (0) (1) (1) (0) (0) (-) (-) (-) (-) (-) (-) minimal ...................................... 0 0 0 1 0 0 - - - - - - severe ....................................... 0 0 1 0 0 0 - - - - - - Dilatation; seminiferous tubules; bilateral .... (0) (0) (1) (1) (0) (0) (-) (-) (-) (-) (-) (-) minimal ...................................... 0 0 1 1 0 0 - - - - - - Sperm granuloma;
unilateral .................... (0) (0) (1) (0) (0) (0) (-) (-) (-) (-) (-) (-) minimal ...................................... 0 0 1 0 0 0 - - - - - - All stages of spermatogenesis normal ........... 10 0 2 10 0 0 - - - - - -
THYMUS;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0) Within Normal Limits............................ 10 0 0 10 0 0 9 0 0 8 0 0 Cyst(s); epithelial ............................ (0) (0) (0) (0) (0) (0) (1) (0) (0) (2) (0) (0) present ...................................... 0 0 0 0 0 0 1 0 0 2 0 0
THYROID GLAND;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
Observations: Non Neo-Plastic ----------------------- MALES ----------------------- ---------------------- FEMALES ----------------------
Removal Reason: TERMINAL SACRIFICE
Group No.: G1 G2 G3 G4 G1R G4R G1 G2 G3 G4 G1R G4R
Dose (mg/kg/day): 0 111 333 1000 0 1000 0 111 333 1000 0 1000
Number of Animals on Study : 10 10 10 10 5 5 10 10 10 10 5 5 Number of Animals Completed: (10) (10) (10) (10) (5) (5) (10) (10) (10) (10) (5) (5)
TONGUE;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
TRACHEA;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
URINARY BLADDER;
Examined........................................ (10) (0) (0) (10) (0) (0) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ 10 0 0 10 0 0 10 0 0 10 0 0
UTERUS;
Examined........................................ (-) (-) (-) (-) (-) (-) (10) (0) (2) (10) (1) (2)
Within Normal Limits............................ - - - - - - 9 0 0 9 0 0 Dilatation; luminal ............................ (-) (-) (-) (-) (-) (-) (1) (0) (2) (1) (1) (2) minimal ...................................... - - - - - - 1 0 0 0 0 0 mild ......................................... - - - - - - 0 0 0 0 0 1 moderate ..................................... - - - - - - 0 0 2 1 1 1
VAGINA;
Examined........................................ (-) (-) (-) (-) (-) (-) (10) (0) (0) (10) (0) (0)
Within Normal Limits............................ - - - - - - 10 0 0 10 0 0
Applicant's summary and conclusion
- Conclusions:
- As there were no treatment-related adverse effects observed up to the highest dose the No Observed Adverse Effect Level (NOAEL) for systemic toxicity of the test item C.I. Pigment Orange 36 is considered to be 1000 mg/kg/day under the test conditions and doses employed
- Executive summary:
The purpose of this repeated dose toxicity study was to evaluate the systemic toxicity profile of the test item, C.I. Pigment Orange 36 in Wistar rats when administered orally by gavage for a period of 90 consecutive daysand to assess the reversibilityof any effects during a subsequent 28days recovery period. This study was also intended to provide the information on major toxic effects, target organs and an estimation of a No Observed Adverse Effect Level (NOAEL).
The test item was weighed and suspended in vehicle,i.e.,0.5% Carboxymethylcellulose Sodium salt (medium viscosity) in Milli-Q®Water and administered to rats at the graduated dose levels of 111, 333 and 1000 mg/kg/day for low dose (G2), mid dose (G3) and high dose (G4 )/ high dose recovery (G4R) group rats, respectively. The rats in the vehicle control group (G1)/vehicle control recovery (G1R) groups received vehicle Carboxymethylcellulose alone. The dose volume administered was 10 mL/kg body weight. Each main group in the experiment was comprised of 10 male and 10 female rats and recovery groups comprised of 5 male and 5 female rats.
The identity of the test item was provided by the Sponsor by a Certificate of Analysis (CoA). The authenticity of the test item was not determined at the test facility. The stability of the test item in the vehicle was established separately under Eurofins Advinus Study No. G19462 at 1 and 100 mg/mL. Based on the results, the test item was found to be stable and homogeneous in the vehicle up to 24 hours when stored at room temperature.
During the conduct of this study, the prepared dose formulations and vehicle (Carboxy methylcellulose Sodium salt (medium viscosity) were analyzed for homogeneity and active ingredient (a.i.) concentration on Day 1 and during 2ndmonth (Day 34) and 3rdmonth (Day 63) of the treatment.The results indicated thatthe percent agreement of the analyzed concentrations were in the range, 85% to 115% of the claimed concentrations and the overall % RSD from six replicates at each dose level was<10.0%. This indicates that the prepared dose formulation met the acceptance criteria for concentration and % RSD.
Each rat in the experiment was observed for clinical signs, mortality and morbidity. Ophthalmological examination was carried out for all the rats prior to start of treatment, at the end of treatment for main groups and at the end of recovery period for recovery groups. The body weights and food consumption were measured during in-life phase of the experiment. Neurological examinations were conducted towards the end of treatment (Day 84) for main groups and towards the end of recovery period (Day 117) for recovery groups.The clinical laboratory investigations such as haematology, coagulation, clinical chemistry, hormone analysis and urine analysis were performed at termination. Vaginal smear was examined in the female rats and the stage ofoestrous cycle was recordedprior to necropsy.
All rats in the experiment were subjected to detailed necropsy and the organ weights and their ratios were derived as percent fasting body weights and brain weight. Histopathological examination was carried out on the preserved organs of the vehicle control (G1) and high dose(G4) group animals. Histopathological examination of the testes included a qualitative assessment of stages of spermatogenesis.In addition, gross lesions from all the animals were examined microscopically.There were no test item-related histopathological changes observed in any organ/tissue in high dose group (G4); hence, histopathological evaluation was not carried out in thelower dose (G2and G3) and recovery groups (G1R and G4R).
Under the experimental conditions employed, the following results were obtained:
· Clinical Signs and Mortality:Orangecolour faecal matter(light to dark) were observed at all the tested doses in both sexes. This could be due to physical nature of the test item. There were no mortality observed at any of the doses tested in both sexes.
· Ophthalmological Examination:Ophthalmological examination did not reveal any ocular abnormalities.
· Neurological Findings:No treatment-related neurological abnormalities /dysfunctions were observed at all the doses tested.
· Body Weights:Treatment did not affect body weight at all the tested doses in either sex.
· Food Consumption:Treatment did not affect food consumption at all the tested doses in either sex.
· Haematology, Coagulation, Clinical Chemistry and urine parameters:There were no test item related alterations observed at any of the tested dose levels in either sex.
· Thyroid Hormone Profile:Thyroid hormone profile (TSH, T4 and T3) was not affected in both sexes across the treated groups when compared to the concurrent vehicle control group.
· Terminal Fasting Body Weights and Organ Weights:No significant changes in terminal fasting body weights and organ weights attributed to test item were observedat any of the tested dose levels in either sex.
· Sperm Parameter:There were no test item-related changes in any of the sperm parameters.
· Gross pathology:There were no test item-related gross pathological changes observed in both sexes. Orange colouration of intestinal contents (ileum, cecum, colon and rectum) observed in both sexes at all doses at the end of treatment period was attributed to the test item colour.
· Histopathology:There were no test item-related microscopic lesions in any evaluated organs or tissues of male and female rats at the end of treatment period at tested dose levels.
No Observed Adverse Effect Level (NOAEL):
As there were no treatment-related adverse effects observed up to the highest dose the No Observed Adverse Effect Level (NOAEL)for systemic toxicityof the test item C.I. Pigment Orange 36 is considered to be 1000 mg/kg/day under the test conditions and doses employed.
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