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Figure Legends
Figure 1.
Insulin resistance indices, serum metabolite concentrations and activities of enzymes,
plasma metabolic hormones, and body condition score (BCS) of prepartum cows during
late gestation (n = 30). The data for plasma insulin and leptin concentrations are weekly
values, whereas those for other parameters are averages of weekly values. Values are
presented as the mean ± SEM. Means with different superscripts (a, b, c) are
significantly different (P < 0.05). ns, not significant. RQ: Revised quantitative insulin
sensitivity check index; RQBHB: Revised quantitative insulin sensitivity check index -
10
β-hydroxybutyrate; HR: Homeostasis model assessment-insulin resistance; Alb:
11
albumin;
12
nonesterified fatty acid; T-Cho: total cholesterol.
AST:
aspartate
aminotransferase;
BHB:
β-hydroxybutyrate;
NEFA:
13
14
Figure 2.
15
Linear regression between IR indices and serum glucose, NEFA and BHB and plasma
16
insulin concentrations of dams (n = 30) during the last 4 weeks of gestation. r;
17
coefficient of correlation. RQ: Revised quantitative insulin sensitivity check index;
18
RQBHB: Revised quantitative insulin sensitivity check index - β-hydroxybutyrate; HR:
19
Homeostasis model assessment-insulin resistance; BHB: β-hydroxybutyrate; NEFA:
20
nonesterified fatty acid.
Table 1. Ingredients and nutrient composition of diets during the
close-up dry period
Item
Close-up dry period
Ingredient composition, kg DM/d
Corn silage
3.0
Grass silage
4.4
Concentrate for dry cows(a)
2.6
Orchard hay
Vitamins, minerals
ad libitum
0.2
Nutrient composition
ME, Mcal/d (% Red)
MP, g/d (% Red)
28.5 (116)
1080.6 (104)
NFC, % DM
32.0
peNDF, % DM
36.2
CP, % DM
13.3
a) Farm aid 18 (nutrient composition on a DM basis: 18.0% CP, 2.0%
crude fat, 10.0% crude fiber, 10.0% crude ash, 0.5% Ca, and 0.4% P;
74.0% TDN, including corn, rapeseed cake, soybean oil cake, corn
gluten feed and corn distiller by-products, bran; Snow Brand Seed Co.
Ltd., Hokkaido, Japan).
DM, Dry matter; ME, Metabolizable energy; MP, Metabolizable
protein; NFC, Non-fibrous carbohydrates; peNDF, Physically effective
Neutral detergent fiber; CP, crude protein.
Table 2. Sequence of primers used for real time PCR (Çolak et al. 2011).
Gene
Primer sequence
GHR
For: 5'–CAC ACC AGC TTT CCT TGT CA-3'
Rev: 5'–GCA GAG ACG ACC ACT TTT GT-3'
For: 5'–GCT GCT GCC TGG GAA TTA-3'
Rev: 5'–CCA TCT GGC TGC CTC TTT-3'
For: 5'–CCA AGG CCA ACC GTG AGA AGA T-3'
Rev: 5'–CCA CGT TCC GTG AGG ATC TTC A-3'
INSR
β-Actin
For, Forward; Rev, Reverse.
GHR, growth hormone receptor; INSR, insulin receptor.
Size (bp)
177
213
256
Table 3. Plasma insulin and serum glucose, NEFA and BHB concentration, and
values of RQ, RQBHB and HR indices of dams in the last 4 weeks of gestation.
Values
Insulin, μIU/mL
Glucose, mg/dL
(mmol/L)
NEFA, mmol/L
BHB, mmol/L
RQ
RQBHB
HR
13.4
69.2
(3.85
0.14
0.42
0.53
0.68
2.26
0.7
0.4
0.02)
0.01
0.01
0.01
0.01
0.12
SD
8.1
4.0
0.22
0.13
0.11
0.07
0.15
1.35
Minimum
2.9
56.0
(3.11)
0.04
0.20
0.36
0.41
0.48
Maximum
43.3
83.6
(4.64)
0.82
0.69
0.69
1.33
7.37
Values; mean ± SEM.
RQ, Revised quantitative insulin sensitivity check index; RQBHB, Revised
quantitative insulin sensitivity check index - β-hydroxybutyrate; HR, Homeostasis
model assessment-insulin resistance; BHB, β-hydroxybutyrate; NEFA, nonesterified
fatty acid.
Table 4. Coefficient of correlation (r) between values of IR indices and the metabolic parameters of dams in the last 4 weeks of gestation.
RQ (n = 30)
RQBHB (n = 30)
HR (n = 30)
-4 wk
-3 wk
-2 wk
-1 wk
-4 wk
-3 wk
-2 wk
-1 wk
-4 wk
-3 wk
-2 wk
-1 wk
0.095
-0.093
-0.175
-0.393*
0.072
-0.085
-0.085
-0.398*
0.261
0.438*
0.075
0.446*
Glucose -0.208
-0.248
-0.275
-0.471**
-0.227
-0.223
-0.275
-0.450*
0.279
0.102
0.118
0.494**
BCS
Insulin
-0.693** -0.496** -0.565** -0.603**
-0.704** -0.582** -0.631** -0.635**
NEFA
-0.318†
-0.391*
-0.387*
-0.525**
-0.243
BHB
-0.449*
0.111
-0.342†
-0.365*
-0.656** -0.343†
Leptin
-0.176
0.149
0.394*
0.572**
-0.107
0.011
0.328†
T-Cho
0.185
0.207
0.361†
0.307†
0.277
0.186
0.353†
AST
-0.420*
-0.013
-0.084
-0.169
-0.425*
-0.019
Alb
-0.318†
-0.459*
-0.351
-0.499**
-0.322†
-0.566** -0.295
-0.243
-0.224
0.995** 0.988** 0.980** 0.995**
-0.483**
-0.388*
-0.463*
-0.688** -0.635**
0.461*
0.337†
0.472*
0.129
0.162
-0.076
-0.295
0.209
-0.066
0.098
-0.125
-0.176
-0.168
0.189
-0.074
0.117
0.014
-0.433*
0.100
-0.162
-0.371*
-0.257
0.597** 0.435*
0.628** 0.212
0.585**
† P < 0.1; * P < 0.05; ** P < 0.01.
RQ, Revised quantitative insulin sensitivity check index; RQBHB, Revised quantitative insulin sensitivity check index - β-hydroxybutyrate;
HR, Homeostasis model assessment-insulin resistance; Alb, albumin; AST, aspartate aminotransferase; BCS, body condition score; BHB,
β-hydroxybutyrate; NEFA, nonesterified fatty acid; T-Cho, total cholesterol.
Table 5. Mean values of metabolic parameters of newborn calves and coefficient of correlation (r) between IR indices of dams in the last 4 weeks of gestation.
Values
RQ (n = 30)
RQBHB (n = 30)
HR (n = 30)
(min-max)
-4 wk
-3 wk
-2 wk
-1 wk
-4 wk
-3 wk
-2 wk
-1 wk
-4 wk
-3 wk
-2 wk
-1 wk
47.3
1.0
BW, kg
0.185
0.012 -0.208
-0.057
0.193
-0.025
-0.196 -0.027
-0.419* -0.329† -0.170
-0.319†
(38.0 - 60.0)
95.8 ± 7.3
0.150
-0.109 -0.074
-0.038
0.199
0.019
-0.111 -0.004
0.121
0.041
0.103
-0.048
Glucose, mg/dL
(38.0 – 239.9)
14.1 ± 2.7
Insulin, μIU/mL
-0.139
0.131 -0.147
-0.197
-0.135
0.214
-0.087 -0.206
0.138
0.044
0.005
0.262
(2.3 – 69.7)
0.79 ± 0.08
0.116
0.000
0.251
-0.039
0.001
0.099
0.108 -0.019
0.062
0.236
0.179
0.033
NEFA, mmol/L
(0.13 – 1.63)
0.04 ± 0.00
BHB, mmol/L
0.152
-0.248 -0.121
-0.112
0.092
-0.245
-0.190 -0.159
-0.029
0.171
0.011
-0.176
(0.03 – 0.07)
20.3 ± 1.1
T-Cho, mg/dL
-0.061
-0.418* -0.202
-0.166
0.036
-0.324† -0.120 -0.142
-0.297
-0.053 -0.226
-0.069
(11.0 – 35.6)
32.4 ± 2.6
AST, IU/L
-0.075
-0.223 -0.355† -0.464*
-0.159
-0.220
-0.407* -0.537**
-0.056
0.106
0.193
0.213
(17.0 - 70.0)
3.0 ± 0.0
Alb, g/dL
-0.150
-0.038 -0.102
0.003
-0.066
0.003
-0.088 -0.051
-0.088
-0.055 -0.301
-0.142
(2.7 – 3.3)
15.3 ± 1.6
GH(a), ng/mL
0.251
0.047
0.290
0.358†
0.290
-0.051
0.425* 0.399*
-0.168
-0.070 -0.218
-0.115
(5.1 - 46.4)
70.6 ± 5.5
IGF-1(a), ng/mL
0.122
0.259
0.220
0.163
0.187
0.080
0.303
0.135
-0.013
-0.050 -0.192
0.125
(35.0 - 162.6)
1×1011 ± 6×1010
-0.164
-0.509 -0.309
-0.773**
-0.355
-0.609* -0.491 -0.736**
0.464
0.900** 0.418
0.727**
Muscle-GHR(b)
(9×10-2 - 6×1011)
3×10-8 ± 1×10-8
Muscle-INSR(b)
-0.245
-0.327 -0.091
-0.545†
-0.373
-0.455
-0.327 -0.591†
0.609* 0.855** 0.436
0.664*
(2×10-9 - 1×10-7)
8×1024 ± 7×1024
Liver-GHR(c)
-0.310
-0.429 -0.619† -0.714*
-0.286
-0.429
-0.667† -0.786*
0.857** 0.833** 0.619† 0.619†
(8×10-1 - 5×1025)
5×10-7 ± 1×10-7
Liver-INSR(c)
-0.152
0.233 -0.170
-0.462
-0.143
-0.224
-0.382 -0.694†
0.690* 0.548
0.190
0.476
(2×10-8 - 9×10-7)
Mean ± SEM., † P < 0.1; * P < 0.05; ** P < 0.01., (a) n = 29. (b) n = 11. (c) n = 8.
RQ, Revised quantitative insulin sensitivity check index; RQBHB, Revised quantitative insulin sensitivity check index - β-hydroxybutyrate; HR, Homeostasis model
assessment-insulin resistance; Alb, albumin; AST, aspartate aminotransferase; BHB, β-hydroxybutyrate; BW, body weight; GH, growth hormone; GHR, growth
hormone receptor; IGF-1, insulin-like growth factor 1; INSR, insulin receptor; NEFA, nonesterified fatty acid; T-Cho, total cholesterol.
...
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