Chinese Meishan pigs produce three to five more pigs per litter than less-prolific U.S. or European pig breeds as a result of a markedly decreased placental size and an increased pig weight: placental weight ratio (placental efficiency). We hypothesized that as a result of their intense selection for prolificacy, the Chinese had indirectly selected for a smaller, more efficient placenta in the Meishan breed. The goals of this study were to determine whether 1) significant variation in placental size and efficiency existed within our population of purebred Yorkshire pigs and 2) selection of pigs (boars and gilts) based on clear differences in placental size and efficiency would affect litter size. There was significant (approximately threefold) variation in placental efficiency in our herd of Yorkshire pigs, and marked (approximately twofold) variation existed within individual litters. We then selected pigs (boars and gilts) that had either a higher (A Group) or lower (B Group) than average placental efficiency. Although the birth weights of selected A Group pigs were similar to those of the B Group pigs, they had markedly smaller placentae. Males from each group (A or B) were bred to the females of the same group, and farrowing data were collected from parities 1 and 2. In both parities, A Group females farrowed more live pigs per litter than did B Group females (12.5 +/- .7 vs 9.6 +/- .5, P < .05). Although A Group pigs were on average approximately 20% lighter than B group pigs (1.2 +/- .1 vs 1.5 +/- .1 kg, P < .05), their placentae were approximately 40% lighter (250 +/- 10 vs 347 +/- 15 g, P < .01), resulting in a marked increase in placental efficiency. The results of this study suggest that selection on placental size and efficiency may provide a valuable tool for optimizing litter size in commercially important pig breeds.
To investigate the impacts of uterine type and conceptus genotype on development through late gestation, Meishan and Yorkshire embryos were co-transferred into the uteri of either Meishan or Yorkshire recipients that were subsequently slaughtered on Day 90 of gestation. At slaughter, regardless of conceptus genotype, fetuses and placentae were markedly smaller when recovered from Meishan than from Yorkshire recipients. Whereas Meishan and Yorkshire fetuses recovered from Meishan uteri were similar in weight, Meishan fetuses were markedly lighter than littermate Yorkshire fetuses when recovered from Yorkshire uteri. Because of the marked differences between fetal weights observed in Yorkshire recipients on Day 90 of gestation, Meishan and Yorkshire embryos were co-transferred to Yorkshire recipients that were allowed to farrow. Surprisingly, Meishan and Yorkshire fetuses cogestated in Yorkshire recipients were born at similar weights, whereas Meishan placentae were markedly smaller. The weight of Meishan placentae were similar on Day 90 and at term, whereas the weight of Yorkshire placentae were markedly larger (approximately 70%) at term than on Day 90. The constant weight of Meishan placentae from Day 90 to farrowing appears to result from an ability to increase their vascularity during this interval. In contrast, Yorkshire placentae may be forced to increase their weight to keep pace with fetal growth during this period. Because uterine capacity sets the upper limit on litter size, the decreased endometrial surface area required per conceptus in the Meishan pig seems to explain its greater potential for increased litter size.
Straightbred Yorkshire (Y) conceptuses are larger than straightbred Meishan (M) conceptuses throughout gestation and at farrowing. In contrast, when Y and M conceptuses were gestated together in Y recipient females, the birth weight of M pigs was similar to that of their Y littermates. Even though placentae of M pigs remained markedly smaller than placentae of Y littermates, they were significantly more vascular. The objective of this study was to compare and contrast the changes in Y and M conceptus growth and placental-endometrial vascularity throughout late gestation in Y or M uteri. Gravid uteri were recovered at slaughter from M and Y females that were gestating either M or Y conceptuses on d 70, 90, or 110 of gestation. Uterine and conceptus measurements were recorded, and a section of the intact endometrial-placental attachment site for each conceptus was fixed, embedded, and later evaluated for placental and endometrial vascular density. Placental surface area and weight were greater (P < .001) when M or Y conceptuses were recovered from Y uteri compared with M uteri on each day of gestation examined. Further, by d 110, the surface area of Y placentae was greater (P < .001) than that of M placentae, regardless of uterine type in which they were gestated. The vascular density of M placentae and adjacent endometrium doubled (P < .05) between d 70 and 110 of gestation (3.0 and 2.5 vs 6.0 and 5.1%, respectively), with no significant increase in placental surface area. In contrast, the surface area of Y placentae doubled in size (P < .001) between d 90 and 110 of gestation, but placental and adjacent endometrial vascular density remained relatively constant, averaging 3.2 and 3.8%, respectively. These data are consistent with the premise that placental size is largely determined by the uterus in which a conceptus is gestated until approximately d 90. After d 90, fetal breed-specific mechanisms maintain optimal fetal growth. Between d 90 and term, M fetal growth depends on progressive increases in placental blood vessel density and requires no increase in placental size. In contrast, Y conceptuses seem to rely exclusively on placental growth to increase placental-endometrial surface area for nutrient exchange.
Agents have been designed and synthesized which target the dimerization interface of HIV-1 protease. These agents, which contain cross-linked peptides from the N- and C-termini of the protease, both inhibit HIV-1 protease activity and decrease the amount of protease dimer in solution as measured by size exclusion chromatography, protein crosslinking, and protease fluorescence studies. Additionally we have shown that active site-targeted agents inhibit HIV-1 protease activity but have little effect on protease dimerization. These data support the claim that inhibition with the crosslinked agents is based on a decrease in the amount of protease homodimer in solution which in turn is responsible for a decrease in the activity of the protease.
Embryonic and fetal mortality reduce lambing rates and litter sizes, thus contributing to economic losses in the sheep industry. In the current study, the timing of late embryonic and fetal loss in ewes and the factors with which these losses were associated were examined. Ewes lambing and lambs born were compared with pregnancy diagnosis and counts of embryos by ultrasonography near d 25, 45, 65, or 85 of gestation. Approximately 19.9% of the ewes experienced late embryonic loss, fetal loss, or both; and 21.2% of the embryos or fetuses were lost from d 25 to term. Potential offspring were lost throughout gestation; 3.7% of embryos from d 25 to 45, 4.3% of fetuses from d 45 to 65, 3.3% from d 65 to 85, and 11.5% from d 85 to parturition; thus, approximately 3 to 4% of the potential offspring were lost for each 20-d period of pregnancy beyond d 25. A greater proportion of ewes lost one (36.7%) rather than all (20.5% single; 3.8% multiple) embryos or fetuses. The patterns of loss were similar in ewes mated during the anestrous season and the transitional period and did not vary with service period within breeding season or method of synchronization of estrus. Late embryonic or fetal losses were not related to the temperature-humidity index. Maternal serum collected near d 25, 45, 65, or 85 of gestation was assayed for concentrations of progesterone, estradiol-17beta , and vascular endothelial growth factor (VEGF). The proportions of embryos or fetuses lost were associated with breed type (P < 0.05), as were concentrations of progesterone (P < 0.01), estradiol (P < 0.05), and VEGF (P < 0.01). The relationships of loss or retention of pregnancy to hormonal variables at the 4 stages studied were limited. Complete and partial losses increased rapidly as maternal progesterone at d 25 decreased below 2 ng/mL (P < 0.05). Survival of fetuses within a litter from d 25 to 65 was greater for ewes with medium concentrations of VEGF near d 25 and from d 65 to parturition was greater for ewes with high concentrations of VEGF near d 45 (P < 0.05). In summary, late embryonic or fetal losses occurred from d 25 throughout gestation and varied with breed type and with concentrations of progesterone in maternal serum on d 25.
We investigated the temporal association between placental vascular endothelial growth factor (VEGF), a potent stimulator of angiogenesis and vascular permeability, and changes in placental/endometrial vascularity on selected days throughout gestation in the pig. Placental and endometrial tissues were collected from sows on Days 25 (n = 4), 36 (n =6), 44 (n = 6), 70 (n =5), 90 (n =5 ), and 112 (n = 7) of gestation. Cross sections of the placental/endometrial interface of each conceptus were used to estimate the number of blood vessels per unit area via image analysis and the intensity of VEGF staining via immunohistochemistry. Placental tissues were also collected on these days to evaluate VEGF mRNA expression. Placental VEGF mRNA expression and the numbers of blood vessels per unit area of placental and adjacent endometrial tissue were low and decreasing from Day 25 to Day 44, before increasing (P < 0.05) markedly and progressively through Day 112. These data are consistent with the marked increase in VEGF immunostaining in the chorionic and uterine luminal epithelium from early to late gestation. Further, these increases in placental VEGF mRNA were positively correlated with fetal weight (r = 0.73; P < 0.0001) and placental efficiency (fetal weight/placental weight ratio; r = 0.66, P < 0.0001). These data are consistent with a role for VEGF in increasing the number of blood vessels at the placental endometrial interface, resulting in an increased capacity for nutrient transfer from the maternal to the fetal compartment.
High proportions of embryonic and early fetal losses in dairy cattle are associated with low peripheral concentrations of progesterone, which could result from increased catabolism, decreased production, or both. Progesterone catabolism occurs primarily in the liver via the cytochrome P450 2C (CYP2C) and cytochrome P450 3A (CYP3A) subfamilies (EC 1.14.14.1; unspecific monooxygenases). Recent observations from our laboratory have shown that the fractional rate constant of progesterone decay can be dramatically reduced by insulin because of a decrease in hepatic CYP2C and CYP3A activity. Little information exists on the regulation of progesterone catabolic enzymes in dairy cows. We hypothesized that elevated insulin concentrations would down-regulate hepatic CYP2C and CYP3A mRNA; therefore, our objectives were to determine the relative abundance of hepatic CYP2C and CYP3A mRNA in dairy cows in response to elevated concentrations of insulin. In the first experiment, 17 mature Holstein cows were drenched daily with 500 mL of water (n = 10) or propylene glycol (a gluconeogenic substrate; n = 7) from 10 d before their expected calving date until d 25 postpartum. Cows drenched with propylene glycol had a 30% increase in peripheral concentrations of insulin. Liver biopsies were collected on d 25 postpartum to determine the relative abundance of CYP2C and CYP3A mRNA. In the second experiment, 19 mature, lactating Holstein cows were randomly assigned to a hyperinsulinemic-euglycemic clamp (0.3 or 1.0 microg of insulin/kg of BW per h; n = 6 each) or remained as controls (saline infused; n = 7) for 96 h beginning on d 10 postpartum. Insulin infusion resulted in a 2.6- or 8- fold increase in peripheral concentrations of insulin, respectively. On d 14 postpartum, a liver biopsy was collected to determine CYP2C and CYP3A mRNA abundance. In experiment 1, the relative abundance of CYP2C mRNA in cows treated with propylene glycol did not differ from controls; however, the relative abundance of CYP3A mRNA in the propylene glycol group was 63% that of controls. For experiment 2, there was a dose-dependent decrease in the relative abundance of both CYP2C and CYP3A mRNA with increasing dosage of insulin. In conclusion, this study demonstrated that, in the cow, either providing a gluconeogenic feed-stuff or treatment with insulin decreased the abundance of mRNA for enzymes responsible for hepatic progesterone catabolism.
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