Maternal endocrine adaptation throughout pregnancy to nutrient manipulation: consequences for sexually dimorphic programming of thyroid hormones and development of their progeny.

Maternal nutrient restriction during critical windows of fetal development alters postnatal growth, often in a sexually dimorphic manner. Intrauterine growth restriction is frequently characterized by accelerated growth and increased adiposity in later life. Thyroid hormones are implicated as part of the mechanism involved in this scenario via their actions within the hypothalamic-pituitary-thyroid axis. We fed high (H = 240%) and low (L = 70%) levels of recommended daily crude protein intake during the first and second trimesters of gestation to beef heifers to investigate effects to their progeny's plasma concentrations of free and total triiodothyronine (FT3 and TT3) and thyroxine (FT4 and TT4) from birth until weaning at 191 days of age (n = 68). The study design was a two-by-two factorial. For male progeny, exposure to maternal diets low in protein during the first trimester of gestation resulted in greater FT4 at birth (P < 0.05) which was subsequent to lower concentrations of leptin in maternal plasma at 271 days of gestation compared with their high-protein-exposed counterparts. These same animals went on to have greater milk intake during the latter half of the lactation period (P < 0.05) and exhibited faster rates of average daily gain (ADG) relative to birth weight during this time (P < 0.05). For all progeny, independent of sex, exposure to low-protein maternal diets during the second trimester of gestation resulted in greater FT3 relative to TT3 at birth. Because FT3 at birth and 29 days was positively associated with ADG (P < 0.05) and ADG relative to birth weight (P < 0.05), it is proposed that FT3 plays an integral role in catch-up growth in the bovine as per other species. Protein intake during the first and second trimesters of gestation has a sexually dimorphic effect on progeny plasma thyroid hormone concentrations, and these changes are associated with altered milk intake and postnatal growth pathway.

Protein intake during gestation affects postnatal bovine skeletal muscle growth and relative expression of IGF1, IGF1R, IGF2 and IGF2R.

Expression of insulin-like growth factor (IGF)1 and IGF2 and their receptor (IGF1R and IGF2R) mRNA in fetal skeletal muscle are changed by variations in maternal nutrient intake. The persistence of these effects into postnatal life and their association with phenotype in beef cattle is unknown. Here we report that the cross-sectional areas of longissimus dorsi and semitendinosus (ST) muscles were greater for mature male progeny born to heifers fed low protein diets (70% vs. 240% of recommended) during the first trimester. In ST, this was accompanied by greater IGF1, IGF2 and IGF2R mRNA at 680 d. Females exposed to low protein diets during the first trimester had decreased IGF2 mRNA in ST at 680 d, however this did not result in an effect to phenotype. Exposure to low protein diets during the second trimester increased IGF1R mRNA in ST of all progeny at 680 d. Changes to expression of IGF genes in progeny skeletal muscle resulting from variations to maternal protein intake during gestation may have permanent and sex-specific effect on postnatal skeletal muscle growth.

Nutrient intake in the bovine during early and mid-gestation causes sex-specific changes in progeny plasma IGF-I, liveweight, height and carcass traits.

Fetal and postnatal growth are mediated by insulin-like growth factors (IGFs) and their binding proteins (IGFBPs). Maternal nutrient intake during gestation can program the postnatal IGF-axis. This may have significant economic implications for beef cattle production. We investigated the effect of high (H=240%) and low (L=70%) levels of recommended daily crude protein (CP) intake for heifers during the first and second trimesters of gestation in a two-by-two factorial design on progeny (n=68) plasma IGF-I, IGF-II, total IGFBP (tIGFBP), postnatal growth and carcass traits. Calves were heavier at birth following high CP diets during the second trimester (P=0.03) and this persisted to 29d. Plasma IGF-I concentrations of males were greater for HL compared to LL (P<0.01) and HH (P>0.04) from 29 to 657d, and for LH compared to LL from 29 until 379d (P=0.02). Exposure to low CP diets during the first trimester resulted in heavier males from 191d onwards (P=0.04) but a tendency for lighter females from 552d onwards (P=0.07) that had lighter carcass weights (P=0.04). Longissimus dorsi cross-sectional area of all carcasses was greater following exposure to low CP diets during the second trimester (P=0.04). Heifer nutrient intake during the first and second trimesters causes persistent and sex-specific programming of progeny plasma IGF-I, postnatal liveweight and carcass weight. Refining heifer nutritional programs during early gestation may optimize production objectives in progeny.

Dietary manipulation of Bos indicusxheifers during gestation affects the prepubertal reproductive development of their bull calves.

We determined the influence of nutritional protein and energy during the first and second trimesters of pregnancy in composite beef heifers on prepubertal reproductive parameters of their male calves. At artificial insemination, heifers were stratified by weight within each composite genotype into 4 treatment groups: High/High (HH=250% crude protein (CP) and 243% metabolisable energy (ME) for first and 229% CP and 228% ME for second trimester of pregnancy), High/Low (HL=250% CP and 243% ME for first trimester and 63% CP and 176% ME for second trimester) Low/High (LowH=75% CP and 199% ME for first trimester and 229% CP and 228% ME for second trimester) or Low/Low (LL=75% CP and 199% ME for first trimester and 63% CP and 176% ME for second trimester). At 5 months of age, male calves were castrated, and gonadotrophins and testosterone (pre- and post-GnRH challenge), IGF-I and leptin were measured along with testicular parameters. Lower maternal dietary protein and energy levels during gestation were associated with increased prepubertal FSH concentrations (P=0.03) and paired testicular volume (P=0.04) in male offspring. Serum LH (P<0.001) and FSH concentrations (P=0.04) were correlated with seminiferous tubule diameter. Testosterone concentrations were positively correlated with testis measures: paired testicular weight (P<0.001), volume (P=0.03) and seminiferous tubule diameter (P<0.001). Although leptin concentrations were associated with prepubertal age (P=0.04) and body weight (P=0.006), they were not associated with any of the measures of reproductive development, but insulin-like growth factor-I was associated with prepubertal FSH (P=0.005). In conclusion, prepubertal reproductive development of bulls may be affected by prenatal nutrition during early and mid gestation.

Heifer nutrition during early- and mid-pregnancy alters fetal growth trajectory and birth weight.

Maternal nutrient intake during gestation can alter fetal growth. Whilst this has been studied extensively in the sheep, less is known about effects in the bovine. Composite-breed beef heifers were allocated to either a high (H/-=76 MJ metabolisable energy (ME) and 1.4 kg crude protein (CP)) or low (L/-=62 MJ ME and 0.4 kg CP daily) nutritional treatment at artificial insemination. Half of each nutritional group changed to an opposite nutritional group at the end of the first trimester (-/H=82 MJ ME and 1.4 kg CP; -/L=62 MJ ME and 0.4 kg CP daily), resulting in 4 treatment groups: HH (n=16); HL (n=19); LH (n=17); LL (n=19). During the third trimester all heifers were fed the same diets. Fetuses were measured at 4-weekly intervals beginning at day 39 of gestation. Calves were also measured at birth for physical body variables. Low maternal nutrient intake was associated with decreased crown-rump length at day 39 (P<0.01) and increased thoracic diameter at day 95 (P<0.01). Umbilical cord diameter was reduced in L/- fetuses in the first trimester (P<0.05) but was greater in -/L fetuses in the second trimester compared to their respective H counterparts (P<0.05). Calf birth weight was decreased in association with -/L maternal diets (P<0.05). In conclusion, fetal development of cattle may be affected by maternal nutrition as early as day 39 of gestation. This may be followed by either compensatory fetal growth, or alternatively, preferential fetal tissue growth that is dependant upon maternal nutrition. Clearly, calf birth weight may be altered by maternal nutrition during mid-gestation.

Dystocia in 3-year-old beef heifers; relationship to maternal nutrient intake during early- and mid-gestation, pelvic area and hormonal indicators of placental function.

The influence of nutrition during the first and second trimesters of gestation on the occurrence of dystocia was investigated in 3-year-old composite-breed beef heifers. Heifers were allocated according to stratification by weight and genotype to either a high (H/-=76 MJ metabolisable energy (ME) and 1.4 kg crude protein (CP)), or low (L/-=62 MJ ME and 0.4 kg CP daily) nutritional treatment on the day of artificial insemination (day 0) to the same Senepol bull. Half of each nutritional group changed to an opposite nutritional group on day 93 of gestation (-/H=82 MJ ME and 1.4 kg CP; -/L=63 MJ ME and 0.4 kg CP daily), resulting in four treatment groups: HH (n=16); HL (n=19); LH (n=17); LL (n=19). From 180 days until calving all heifers were fed the same diets. Pelvic area measures were taken at heifer selection (-72 days) and at 117 days. Maternal circulating concentrations of estrone sulphate (ES), bovine placental lactogen (bPL), bovine pregnancy associated glycoprotein and progesterone were monitored throughout gestation. Heifers were observed continuously over the calving period and delivery type classified as being either eutocic or dystocic. The occurrence of dystocia was 14.1%. Increased calf birth weight increased the odds of occurrence of dystocia (odds ratio (OR)=1.40; 95% confidence interval (95% CI) 1.12-1.76; P<0.01). High diets in the second trimester were associated with heavier calves at birth (P=0.01). The mean pelvic area of eutocic heifers on -72 d, tended to be greater compared to that of dystocic heifers (P=0.08) such that a 1-cm(2) difference in pelvic area tended to decrease the risk of dystocia (OR=0.97; 95% CI 0.93-1.01; P=0.09). Longer gestation length was associated with an increased risk of dystocia (P=0.03). ES (P=0.04) and bPL (P=0.09) at calving were positively associated with the risk of dystocia. In conclusion, the current study demonstrates (a) that pelvic area measurement at selection in 3-year-old Bos indicusxBos taurus heifers may be useful for identifying heifers at an increased risk of dystocia and (b) increased ES and bPL concentrations at calving are associated with increased risk of dystocia. Pelvic area measurements obtained prior to conception remain valid in their assessment of the relationship between pelvic area and likelihood of dystocia occurring in the event of changing maternal nutrient intake during gestation. This is an important finding given maternal diets high in protein and energy during the second trimester of gestation increased calf birth weight and calf birth weight was associated with an increase in the occurrence of dystocia in heifers calving as 3-year olds.

Influences of diet during gestation on potential postpartum reproductive performance and milk production of beef heifers.

The influences of nutritional protein and energy during early and mid pregnancy on milk production and postpartum reproductive parameters were determined in 70 beef heifers of two composite breeds (Bos indicus X Bos taurus). At artificial insemination (AI), heifers were divided into four dietary treatment groups identified by the level of protein, and to a lesser extent energy, fed during the first and second trimesters: high/high (HH), high/low (HL), low/high (LH), and low/low (LL). Milk production was lower in the heifers receiving high treatment in first trimester than that in heifers receiving the low treatment (P=0.01). Milk production was negatively associated with dam body condition score (BCS; P=0.01), nonesterified fatty acids (P=0.001), and leptin (P=0.02) and positively associated with urea (P<0.001) concentrations during lactation. Increased dietary protein in the first trimester increased or decreased concentrations of colostral protein dependent upon genotype (P=0.03). Colostral protein was positively associated with bovine pregnancy associated glycoprotein from late gestation (P=0.007). Milk fat was negatively associated with BCS (P=0.007) and influenced by genotype (P=0.003). Dietary treatment did not affect the postpartum reproductive performance of beef heifers. Gestation length (P<0.001) and the postpartum interval to first estrus (PPI; P=0.02) were positively associated with calf size. Placental size was negatively associated with placental expulsion time (P<0.01). Prepartum BCS of the heifers was negatively associated with PPI (P=0.01). Overall, high levels of nutrition during early gestation are detrimental to milk production in beef heifers.

Dietary protein during gestation affects maternal insulin-like growth factor, insulin-like growth factor binding protein, leptin concentrations, and fetal growth in heifers.

The influence of supplemental protein during gestation on maternal hormones and fetal growth was determined in composite beef heifers. At AI, 118 heifers were stratified by BW within each composite genotype (BeefX = 1/2 Senepol, 1/4 Brahman, 1/8 Charolais, 1/8 Red Angus and CBX = 1/2 Senepol, 1/4 Brahman, 1/4 Charolais) into 4 treatment groups: high high (HH = 1.4 kg CP/d for first and second trimesters of gestation), high low (HL = 1.4 kg of CP/d for first trimester and 0.4 kg of CP/d for second trimester), low high (lowH = 0.4 kg CP/d for first trimester and 1.4 kg of CP/d and for second trimester), or low low (LL = 0.4 kg CP/d for first and second trimesters). Maternal plasma IGF-I and -II, total IGFBP, and leptin concentrations were determined at 14 d before AI and at d 28, 82, 179, and 271 post-AI (mean gestation length 286 d), and leptin concentrations were also determined at calving. Increased dietary protein increased maternal plasma IGF-I (P < 0.001 on d 28, 82, and 179), IGF-II (P = 0.01 on d 82; P = 0.04 on d 271), and total IGFBP (P = 0.002 on d 82; P = 0.005 on d 179; P = 0.03 on d 271). Maternal plasma IGF-I at d 271 was negatively associated with calf crown-rump length at birth (P = 0.003). BeefX had greater birth weight calves (P = 0.01), greater IGF-II (P < 0.001), increased ratios of IGF-I:total IGFBP (P = 0.008) and IGF-II:total IGFBP (P < 0.001), and reduced total IGFBP compared with CBX (P = 0.02). Increased dietary protein during second trimester increased maternal plasma leptin at calving (P = 0.005). Maternal plasma leptin near term was positively associated with heifer BCS (P = 0.02) and with calf birth weight (P = 0.04), and at calving was positively associated with heifer age at AI (P = 0.02). These findings suggest that maternal dietary protein, age, and genotype influence plasma concentrations of metabolic hormones and fetal growth in Bos indicus-influenced heifers.

Dietary manipulation of Bos indicus x heifers during gestation affects the reproductive development of their heifer calves.

The effect of nutrition during the first and second trimesters of pregnancy in composite beef heifers on reproductive parameters of their female calves was determined in the present study. At artificial insemination, heifers were assigned to one of four treatment groups (i.e. HH, HL, LowH and LL) depending on the level of crude protein intake (H = high; L = low) for first and second trimesters of pregnancy. Gonadotrophin concentrations and ovarian parameters were measured in their female calves at 5 and 23 months of age. Crude protein intake was positively associated with dam plasma urea (P < 0.001). The density of healthy follicles in heifers at the time of death was negatively correlated with dam plasma urea at Day 179 (P = 0.009). Heifers from LowH dams had a smaller-sized prepubertal largest ovarian follicle (P = 0.03) and lower densities of primordial and primary follicles (P = 0.02) and healthy antral follicles (P = 0.009) when they were killed. There was a positive correlation between plasma FSH concentrations at 5 and 23 months of age (P = 0.02), as well as between the sizes of the largest ovarian follicles at 6 and 23 months of age (P = 0.01). In conclusion, the reproductive development of heifers may be affected by prenatal nutrition during early and mid-gestation.

Dietary protein during gestation affects placental development in heifers.

The influence of nutritional protein during the first and second trimesters of pregnancy on placental measures at term and caruncle numbers in the uteri of adult offspring was determined in composite beef heifers. At artificial insemination (AI), heifers were divided by weight and composite genotype into four dietary treatment groups, identified by the level of protein components fed during the first and second trimesters: high/high (HH), high/low (HL), low/high (LH), low/low (LL). Expelled placentas were collected and weighed, and cotyledons were dissected, counted, weighed, and measured. Uteri from mature female offspring were dissected at slaughter and caruncles counted. The number of cotyledons in the expelled placenta was increased by high dietary protein in the second trimester (P=0.02) and varied with genotype (P=0.03). Placental weight was influenced by maternal undernutrition during early gestation dependent on dam genotype (P=0.001). Placental efficiency, as determined by calf weight:placental weight, increased with dam age (P=0.03). Calf birth weight was closely associated with placental weight (P=0.002) and cotyledonary weight (P=0.001) and surface area (P=0.04), but not with the number of cotyledons. Leptin concentrations during early (R=-0.29) and late gestation (R=-0.25) correlated with placental weight, and Insulin-like growth factor binding proteins throughout gestation correlated with the number of cotyledons (R=-0.28 to-0.33). The number of uterine caruncles in the nonpregnant adult offspring did not correlate with the dam's genotype, nutrition treatment, or cotyledon number in the expelled placenta.

Dietary protein during gestation affects circulating indicators of placental function and fetal development in heifers.

The influences of nutritional protein during the first and second trimesters of pregnancy on placental hormones and fetal growth were determined in composite beef heifers. At artificial insemination, heifers were stratified by weight within each composite genotype into 4 treatment groups: High High (HH=1.4kg crude protein (CP)/day for first and second trimesters of gestation; n=16), High Low (HL=1.4kg CP/day for first trimester and 0.4kg CP/day for second trimester; n=19), Low High (LH=0.4kg CP/day for first trimester and 1.4kg CP/day for second trimester; n=17) or Low Low (LL=0.4kg CP/day for first and second trimesters; n=19). Maternal plasma bovine pregnancy associated glycoprotein (bPAG) and progesterone (P4) were determined at gestation day (gd) 28, 82, 179 and 271 (mean gestation length 286 days) in addition to P4 at term. Estrone sulphate (ES) and bovine placental lactogen (bPL) concentrations were measured at gd 124, 179, 236 and 271 and at term in addition to ES at gd 82. Low dietary protein increased placental function as indicated by increased bPAG (P<0.001) and ES (P=0.02) concentrations in first trimester and increased bPL concentrations (P=0.01) in the second trimester of gestation. In the third trimester, when dietary treatment had ceased, placental function was no longer associated with previous dietary treatments. Dam genotype affected placental function as measured by bPL (P<0.001) and ES concentrations (P=0.02). Calf gender, heifer age and maternal insulin-like growth factor (IGF)-I, -II and leptin did not affect hormonal indicators or circulating markers of placental function. Enhanced placental function during the third trimester, as measured by ES, was associated with increased calf birth weight (P=0.003).

Factorial design of electrolyte systems for the separation of fatty acids by capillary electrophoresis.

In this work, a capillary zone electrophoretic methodology using UV indirect detection (224 nm) for the analysis of fatty acids (FAs) in saponified oils is proposed. The electrolyte consisted of a 5 mmol l(-1) phosphate buffer, pH 7. containing 4 mmol l(-1) sodium dodecylbenzenesulfonate (SDBS) as chromophore, 4 mmol l(-1) dimethyl-beta-cyclodextrin and 45% acetonitrile (ACN). The composition of the electrolyte was optimized by a 2(3) factorial design with triplicate at the central point. The design established practical concentration boundaries for SDBS and ACN. In a defined concentration range of 2-4 l(-1), SDBS can certainly be used as a chromophore for indirect detection without imparting excessive baseline noise. For ACN, a suitable interval of 45-55% was found to enhance FAs solubilization without overflowing the system with bubble formation and current interruption. Additionally, the design revealed the importance of dimethyl-beta-cyclodextrin in the resolution of difficult pairs and its function as a solubilizing agent for long chain FAs. At the optimized conditions, nine FAs from C10 to C20, including mono- di- and tri-unsaturated C18 fatty acids were baseline separated in less than 10 min. The proposed method was applied to the separation of FAs in edible oils and polyunsaturated fatty acid enriched margarine. Additionally, spectral monitoring at 206 nm was used to confirm peak identity in the samples.

Captopril in acute myocardial infarction: beneficial effects on infarct size and arrhythmias.

It is known from experiments that angiotensin-converting enzyme inhibitors can limit infarct size. In a prospective, randomized, placebo-controlled double-blind study, 22 patients were given 1.5-2.0 mg captopril/h i.v., while 24 patients were given placebo. Medication was started between 2 and 18 h from the onset of infarction. The two groups were matched for age, infarct location, and time of intervention. With the exception of one patient in either group, all were concurrently given nitroglycerin. The necrosis parameters were provided by the quantitative measurement of the QRS complex. The Q wave decreased with captopril treatment (-0.003 mV), but increased with placebo (+0.14 mV, p < 0.05). The number of ventricular premature beats at 24 h from the start of treatment was 25/h with placebo, and 9/h with captopril (p < 0.02). Ventricular fibrillation occurred seven times in the placebo group, but did not occur in the captopril group. The creatine kinase infarct weight was 59 gram-equivalents (gEq) with placebo, and 45 gEq with captopril (p = NS). Mean arterial pressure was reduced by 12 mmHg with captopril treatment. The results show a beneficial effect of captopril on infarct size and electrical instability, over and above the effect of standard management with nitroglycerin and thrombolysis.

Captopril in acute myocardial infarction. Beneficial effects on infarct size and arrhythmias.

OBJECTIVE: It is known from experiments that angiotensin converting enzyme (ACE) inhibitors can limit infarct size. We examined the effect in patients. METHODS: In a prospective, randomized, placebo-controlled double blind study, 22 patients were given 1.5-2.0 mg captopril/h i.v., while 24 patients were given placebo. Medication was started between 2 hours and 18 hours from the onset of infarction. The two groups were matched for age, infarct location, and time of intervention. With exception of one patient in either group, all were concurrently given nitroglycerin. The necrosis parameters were provided by the quantitative measurement of the QRS complex. RESULTS: The Q wave decreased with captopril treatment (-0.003 mV), but increased with placebo (+0.14 mV) (p < 0.05). The number of ventricular premature beats at 24 hours from the start of treatment was 25/h with placebo, and 9/h with captopril (p < 0.02). Ventricular fibrillation occurred 7 times in the placebo group, but did not occur in the captopril group. The creatine kinase (CK) infarct weight was 59 gram-equivalents (gEq) with placebo, and 45 gEq with captopril (p = NS). The mean arterial pressure was reduced by 12 mmHg with captopril treatment. CONCLUSIONS: The results show a beneficial effect of captopril on infarct size and electrical instability, over and above the effect of standard management with nitroglycerin and thrombolysis.

[Captopril in acute myocardial infarct: its effect on infarct size and arrhythmias]. / Captopril bei akutem Herzinfarkt: Einfluss auf Infarktgrösse und Rhythmusstörungen.

The effect of captopril on infarct size and arrhythmias was determined in a prospective, randomized, placebo-controlled double-blind study of 46 patients (9 women, 37 men; mean age 61 [38-86] years). Within 2-18 hours of entry into the study these patients received either a slow intravenous bolus injection of 2.5 or 5.0 mg captopril followed by a continuous infusion of 1.5-2.0 mg/h for a period of 48 hours (n = 22), or of a placebo by the same mode of administration (n = 24). The two groups were comparable as to age, infarct site and time of intervention. All patients, except one in each group, also received nitroglycerin (1.2-6.0 mg/h intravenously). QRS complexes were measured to provide a necrosis index. Q-wave amplitudes decreased under captopril (-0.08 +/- 0.04 mV) while increasing under placebo (+0.15 +/- 0.04 mV; P less than 0.05). The number of ventricular extrasystoles in the first 24 hours after onset of treatment or on placebo was 25/h and 9/h, respectively (P less than 0.02). Ventricular fibrillation occurred in 7 patients of the placebo group, in none of the captopril group. Creatine-kinase infarct weight was 59 and 45 gram-equivalents, respectively (placebo vs treated group: not significant). The mean arterial blood pressure fell by 14 mm Hg during the first hour in the captopril group, but by only 3 mm Hg on placebo (P less than 0.01). These results indicate that captopril has a favourable influence on infarct size and electrical stability which is additional to that provided by standard nitroglycerin treatment.