Action of GnRH on steroid secretion by luteal cells of cyclic and early pregnant sows in vitro

The effect of GnRH was studied on progesterone (P4), oestradiol-17 beta (E2) and testosterone (T) secretion by porcine luteal cells from the 13th day of the oestrous cycle and the 18th day of pregnancy. Trypsin-dispersed luteal cells (5 X 10(4) cells/ml) were incubated in medium 199 with 10% calf serum with or without GnRH in doses of 0.1, 1, 10 and 100 mg/ml and with 1 microgram LH and 50 U/ml hCG. The concentration of P4, E2 and T in the medium was estimated by radioimmunological method after 6 hours of incubation. The results showed that GnRH had no effect on the secretion of the investigated steroid hormones by luteal cells from cyclic sows. GnRH at a dose of 10 g inhibited E2 secretion and at a dose of 1 ng T secretion by cells from pregnant sows. LH and hCG stimulated release of P4 by luteal cells in both physiological stages. The conclusion drawn was that GnRH does not act directly on luteal cells of cyclic sows but may inhibit E2 and T secretion by cells of pregnant sows.     

Synthesis and secretion of immunoreactive methionine-enkephalin from rabbit reproductive tissues in vivo and in vitro.

The expression of the proenkephalin gene has been demonstrated in the reproductive tissues of several animal species. The objectives of the experiments reported here were to (a) examine the presence of immunoreactive methionine-enkephalin (ir-MENK) in rabbit ovary, oviduct, and uterus and in a rabbit endometrial cell line (HRE-H9), (b) characterize ir-MENK biochemically, (c) investigate the effect of eCG + hCG treatment on the synthesis and secretion of ir-MENK in vivo, and (d) study the effect of K+ depolarization on the secretion of ir-MENK from HRE-H9 cells. Uterine fluid was collected by flushing the uterine lumen with saline. Reproductive tissues and HRE-H9 cells were extracted with 0.1 N acetic acid. Both the uterine fluid and extracts of uterus, ovary, oviduct, and HRE-H9 cells exhibited inhibition curves parallel to that of authentic MENK in the MENK RIA system. Sephadex G-15 gel filtration profiles indicated that in the extracts of rabbit uterus and HRE-H9 cells, most ir-MENK co-eluted with standard MENK, with a minor portion eluting near the void volume (Vo). Reverse-phase-HPLC (RP-HPLC) profiles showed a major peak coinciding with standard MENK, plus a minor peak of highly hydrophilic ir-MENK. The effect of eCG + hCG treatment was studied by i.m. injection of eCG (150 IU), followed by i.v. injection of hCG (75 IU) 4 days later. Ir-MENK concentration in the uteri and ovaries was significantly (p less than 0.05) increased (9.06 +/- 1.89 and 2.05 +/- 0.32 ng/mg protein, respectively), compared to control levels (2.31 +/- 0.86 and 0.24 +/- 0.77).(ABSTRACT TRUNCATED AT 250 WORDS)     

Factors affecting ice nucleation in plant tissues

Factors affecting the ice nucleation temperature of plants and plant tissues were examined. The mass of a sample had a marked effect on ice nucleation temperature. Small tissue samples supercooled to −10°C and were not accurate predictors of the nucleation temperature of intact plants in either laboratory or field experiments. This effect was not unique to plant tissues and was observed in autoclaved and control soil samples. Ice nucleation temperatures of bean, corn, cotton, and soybean seedlings were influenced by the length of subzero exposure, presence of ice nucleation active bacteria, and leaf surface wetness. The number of factors influencing ice nucleation temperature suggested that predicting the freezing behavior of plants in the field will be complex.     

Prolactin stimulates and potentiates adrenal steroid secretion in vitro

Prolactin, alone and in combination with ACTH, was tested for its ability to release steroids from rat adrenocortical slices superfused in vitro. The hormone possessed weak activity alone (minimal responsive dose = 1 U), but was able to potentiate the ACTH-stimulated corticoid release at much lower doses (significant response over ACTH alone at 0.01 U prolactin). This latter dosage was calculated to be within the physiologic range of prolactin in blood. Analysis of individual steroids in superfusate by RIA revealed that aldosterone release was most sensitive to prolactin, followed by corticosterone, androstenedione, and progesterone. We conclude that prolactin is an adrenocortical secretagogue of physiological relevance in the rat, and that it could play a role in enhancing the action of ACTH to acutely release steroids.     

Factors affecting reproductive performance of Holstein heifers

The objectives were to evaluate factors affecting reproductive performance of dairy heifers. Holstein heifers (6389) were housed in a feed lot located in Parma, ID. Each week heifers weighing > or =290 kg were initiated in the reproductive program, which consisted of one injection of PGF(2alpha) and AI on detection of estrus. Heifers not inseminated by 11 days after the initiation of the breeding program received a second injection of PGF(2alpha). Pregnancy was diagnosed at 40+/-3 and 90+/-3 days after AI. Average daily minimum temperature (ADMnT), average daily maximum temperature (ADMxT), and average daily rainfall (ARF) were recorded between 15 days prior to and 15 days after the day of AI or the day of initiation of the breeding program. Exposure to air temperature was classified as: cold stress (CS=ADMnT< or =4 degrees C), no stress (NS=ADMnT>4 degrees C and ADMxT<29 degrees C), and heat stress (HS=ADMxT> or =29 degrees C). Exposure to rainfall was classified as above (HRF) or below (LRF) the mean for the period in question. Heifers were classified according to body weight at initiation of the breeding program as thin (TH<340 kg); moderate (MD=340-365 kg); and heavy (HY>365 kg). Service sire was associated with conception rate at 40 and 90 days after first AI. Although exposure to air temperature was not correlated with conception rate at 40 days after first AI, heifers exposed to cold stress had smaller conception rates at 90 days after first AI because they were more likely to lose pregnancy between 40 and 90 days of gestation. The proportion of heifers inseminated after initiation of the breeding program was correlated with body weight and exposure to cold stress. Exposure to cold stress was also correlated with the proportion of heifers conceiving within 11 and 22 days after initiation of the breeding program. From this study a correlation was established between body weight and rate of insemination and between the exposure to cold stress and reproductive efficiency of Holstein heifers.     

Factors affecting reproductive performance in captive Mallard ducks

Propagation of wild-strain Mallards (Anas platyrhynchos) in captivity is hindered by low egg fertility. Therefore, we studied the effects of captivity, age, mate choice and isolation on reproductive parameters of Mallards. Captive drakes had smaller immature testes than free-flying Mallard drakes. Captive yearling ducks weighed less than adult ducks at the beginning of the breeding season, but no differences were found between their initial clutch size, egg volume or number of clutches laid. Yearling pairs had lower egg fertility (7%) than adult pairs (80%). Egg fertility was higher (51 vs 21%) in self-chosen pairs than in randomly assigned pairs. Isolation of ducks, however did not influence egg production.     

Factors affecting reproductive performance in Ontario dairy herds

Data were collected on reproductive, health and production events in 45 dairy herds in southwestern and eastern Ontario, Canada, from July 1990 to July 1993. Mean and median days in milk at first estrus, at first service, and at conception were 85.1 and 83.9, 94.0 and 88.9, and 126.3 and 115.5 d, respectively. The mean estrus detection rate was 48.3%, with a minimum of 28.4% and a maximum of 64.2%. The mean overall conception rate was 46.7%, with a minimum of 29.8% and a maximum of 70.7%. The most commonly used used therapeutic agents were GnRH (11.5% lactational incidence risk) and PG (13.1% lactational incidence risk). Estrus management accounted for the largest portion of PG use in the herds studied (69.4%). Results of a two sample t-test indicated no significant difference in reproductive performance among herds housed in free-stall compared with those housed in tie-stall barns. Multivariate linear regression models of 3 reproductive performance measures indicated that cows with ovarian cysts and/or increased peak milk production had significantly longer calving-to-first estrus and calving-to-conception intervals as well as lower conception rates than average producing cows without cysts. Cows with metritis were found to have significantly shorter calving-to-conception intervals but no detectable difference in conception rate or calving-to-first estrus interval than their herdmates. As lactation number increased reproductive performance improved. Cows that were observed in estrus earlier conceived earlier. Animals receiving uterine infusions had significantly longer calving-to-conception intervals and lower conception rates than their herdmates. There was no detectable difference in reproductive performance for cows treated with GnRH or PG compared with the nontreated cows. An increased estrus detection rate was associated with shorter calving-to-conception intervals and lower conception rates. Based on the range of performance in the study herds, the factors with the greatest potential influence on calving-to-conception interval in the average herd were the estrus detection rate and conception rate.     

Factors affecting the initiation of mini-rhizomes from Trillium erectum and T. grandiflorum tissues in vitro

Leaf and stem explants of Trillium grandiflorum and T. erectum produced mini-rhizomes (MRs) in vitro which gave rise to shoots and roots. The apical portion of the stem and the basal portion of the leaves were the most effective explants from these tissues, while stem tissue was more responsive than leaf tissue. The best response with both species was observed on half-strength MS basal medium supplemented with cytokinin and auxin. T. erectum was more responsive than T. grandiflorum overall, and in some cases produced MRs in the absence of growth regulators. Culture at 21°C appeared to stimulate growth from T. grandiflorum tissues, compared with controls at 27°C, whereas the outgrowth of shoots from MRs was inhibited in both species at 21°C. In vitro production of MRs could provide a more rapid, alternative propagation method for these species than traditional methods.     

LH-RH antagonist inhibits gonadal steroid secretion in vitro

This investigation was aimed at studying the direct action of LH-RH derivatives on gonadal function. Modulation by LH-RH antagonist (Ac-[D-beta-Nal1, D-p-Cl-Phe2, D-Trp3, D-Arg6, D-Ala10]-LH-RH) and agonist of LH-RH (D-Ser(TBU)6, AzaGly10-LH-RH) and native LH-RH of HCG-stimulated steroidogenesis in testicular Leydig cells and luteal cells was studied in vitro. The LH-RH antagonist (3.2 X 10(-8) M) was found to change ED50 of HCG from 2 X 10(-11) M to 5.5 X 10(-11) M in the Leydig cell culture system. In addition, the antagonist was noted to override the stimulatory action of native LH-RH in Leydig cells. Furthermore, the agonist was found to augment HCG-provoked testosterone secretion. Similarly, the LH-RH antagonist at 10(-9) M blunted HCG-stimulated progesterone secretion in the luteal cell culture system and increased the ED50 of HCG from 8.7 X 10(-13) to 7.7 X 10(-12) M. In contrast to the Leydig cell culture system native LH-RH (10(-8) M) and the agonist (10(-8) M) increased the ED50 of HCG in luteal cells from 8.7 X 10(-13) M to 3 X 10(-12) M and 2.3 X 10(-12) M, respectively. Present data combine to suggest that LH-RH antagonists act at least partially at the gonadal level and may be clinically useful to inhibit Leydig cell and ovarian function.     

Ovarian steroid secretion changes after hCG stimulation in early pregnant pigs

Administration of human chorionic gonadotropin (hCG) to promote ovarian steroid secretion near the time of recognition of pregnancy was evaluated. Neither 500 or 1000 IU of hCG caused a significant increase in luteal function as determined by progesterone (P(4)) concentrations in peripheral blood following treatment on Day 12. Estradiol concentrations were elevated (P<0.01) for the 500 IU hCG group on Days 13, 14, 15 and 16 versus the control group. The 1000 IU of hCG group had three-to five-fold greater (P<0.01) estradiol concentrations than controls on Days 14, 15 and 16 post mating. Treatment with hCG also reduced (P<0.05) the number of resorbed embryos. The results suggest that hCG treatment on Day 12 of pregnancy reduced embryo loss and influenced peripheral estradiol secretion patterns.     

Metabolism of steroid hormones in vitro by follicular tissues of the Japanese quail

The cell-free homogenates of the theca layers and granulosa layers of quail follicles were incubated at 39 degrees C with 14C-labeled steroids in the presence of NADPH. At the end of incubation, radioactive steroids were extracted and analyzed by thin-layer chromatography. When radioactive progesterone was employed as the substrate, 17 alpha-hydroxyprogesterone and androstenedione were obtained as the metabolites. 17 alpha-Hydroxylase activity, estimated from the amounts of these two metabolites, was high in the theca layers of the second largest (F2) and the third largest (F3) follicles. The theca layer of the largest follicle (F1) and the granulosa layers of all three follicles were essentially devoid of this enzyme activity. The activity of C17-20 lyase was estimated from the amount of androstenedione that was obtained as a sole metabolite in the incubation of radioactive 17 alpha-hydroxyprogesterone. This enzyme showed a tissue distribution similar to 17 alpha-hydroxylase. When radioactive androstenedione was used as the substrate, testosterone, 5 beta-androstane-3,17-dione, and 3 beta-hydroxy-5 beta-androstan-17-one were identified as the metabolites. 17 beta-Hydroxysteroid dehydrogenase activity, estimated from the amount of testosterone, was higher in the granulosa layers than in the theca layers. On the other hand, 5 beta-reductase activity, estimated from the sum of 5 beta-androstane-3,17-dione and 3 beta-hydroxy-5 beta-androstan-17-one, was almost equally distributed in the two layers. In order to investigate the changes in the enzyme activities during the ovulatory cycle, birds were killed at various times before the predicted ovulation of F1. When the 17 alpha-hydroxylase activity was estimated in the cell-free homogenates of the theca layers, peaks in the activity were observed 32, 42, 54, and 66 h before ovulation of F1. There was a small peak 18 h before ovulation, and activity then started to decrease. The change of C17-20 lyase activity during the cycle was completely parallel with that of 17 alpha-hydroxylase activity.     

Factors influencing the postweaning reproductive performance of sows on commercial farms

The objective of this study was to investigate the effects of various factors, including lactational feed intake, on the reproductive performance of sows in commercial herds. The 4 measures of reproductive performance were weaning-to-first-service interval, weaning-to-conception interval, litter weight at weaning, and subsequent litter size. Parity, farrowing season, lactation length, farrowing-to-conception interval, litter size, and lactation feed intake were investigated as risk factors common to the 4 measures of post-weaning reproductive performance. Using 4 basic multiple regression models for each measure, the least-square means for sets of factors were compared using the GLM procedure of SAS. Parity 1 sows had the longest weaning-to-first-service interval and weaning-to-conception interval, and the lighter litter weight at weaning (P < 0.05) than mid-parity sows. Sows in Parities 2 to 5 had larger subsequent litter size (P < 0.05) than those in Parities 1 and >/= 7. Sows farrowing in summer and spring had the longest and second longest weaning-to-conception interval (P < 0.05), respectively, while sows farrowing in summer had longer weaning-to-first-service interval than those that farrowed in spring (P < 0.05). Sows farrowing in summer produced the lightest litter weight at weaning (P < 0.05). No differences in subsequent litter sizes were found due to farrowing season (P > 0.10). As lactation length increased, weaning-to-first-service interval and weaning-to-conception interval decreased, and litter weaning weight increased. Longer lactation length and farrowing-to-conception interval were associated with larger subsequent litter size (P < 0.05). Litter size did not affect weaning-to-first-service interval or weaning-to-conception interval. Larger litter sizes were associated with heavier litter weight at weaning. Greater lactation feed intake improved the 4 measures of reproductive performance.     

Carbon balances for in vitro digestion and fermentation of potential roughages for pregnant sows

Ad libitum feeding of pregnant sows requires satiating, intake-restricting feed components to prevent sows from getting excessively fat. Because hindgut fermentation starts only after and proceeds much slower than enzymatic digestion in the small intestine, fermentation products might, as nutrients, induce a prolonged physiological satiation of sows. To simulate hindgut fermentation and determine the fermentative release of short-chain fatty acids (SCFA) in vitro, gas production tests (GPTs) were performed with different raw materials after removal of enzymatically hydrolysable compounds. Fresh feces from sows that received standard sow feed was used as inoculum. Fresh, ensiled, or dehydrated by-products of the food industry (brewers’ grains, liquid yeast feed, maize gluten feed, raw potato chips, potato steam peel, pressed potato pulp, sugar beet pulp (SBP)) and whole-plant products (grass, maize) were tested as individual products.

Balances were drawn up of the in vitro flow of organic carbon (OC) to an “ileal” nutrient fraction, a “hindgut” fraction of SCFA and gas, and “fecal” remnants. The OC balances revealed large variations among raw materials in terms of their contribution to the different fractions. Potato steam peel gave the largest “ileal” nutrient fraction (77% of total OC), the lowest was observed with fresh sugar beet pulp (9% of total OC). In the GPT, SBP, and potato pulp brought about the highest “hindgut” SCFA yields (32–49% of total OC), and together with raw potato chips, the highest amounts of gas. Grass products and liquid yeast feed were slower fermented than most by-products. Moreover, whole-plant materials gave larger “fecal” OC portions than by-products, with the exception of fresh and dehydrated brewer’s grains. Together with straw, the latter were the least degradable of all raw materials tested. Among the grass products, dehydrated grass, and among the by-products, raw potato chips left the least “fecal” residues, i.e. they were nutritionally utilized to the largest extent.     

Estrogen sulfatase and steroid sulfatase activities in intrauterine tissues of the pregnant guinea pig

The possible role of intrauterine estrogen sulfatase and steroid sulfatase around the time of parturition in the guinea pig was investigated. [3H]Estrone sulfate or [3H]pregnenolone sulfate was incubated with intrauterine tissues. Estrogen sulfatase was found in placenta, endometrium, decidua basalis, amnion and chorion. The presence of steroid sulfatase was established in endometrium and decidua basalis but not in placenta or the fetal membranes. Examination of activities in early (days 32-35), mid (days 44-46) and late (within 5 days of parturition) gestation revealed no significant change in estrogen sulfatase specific activity in decidua basalis. However, in chorion and endometrium this activity was seen to increase approx. 12-fold (P less than 0.001) and 2.8-fold (P less than 0.001), respectively, from early to late gestation. In placenta, estrogen sulfatase activity appeared to increase 2.4-fold (P less than 0.001) and in amnion it decreased 2.8-fold (P less than 0.002). Steroid sulfatase activity in decidua basalis did not change during gestation, while activity in endometrium was found to increase by a factor of 5.3 (P less than 0.001), from early to late gestation. The increases, both in estrogen sulfatase activity in chorion, endometrium and placenta and in steroid sulfatase activity in endometrium, occurred primarily within the final 3 weeks of gestation. In contrast, the decrease in estrogen sulfatase activity in amnion occurred principally between the fifth and sixth weeks of gestation. Analysis of radiolabelled metabolites indicated that estradiol and progesterone could be produced via estrogen sulfatase and steroid sulfatase activities in certain tissues. Subcellular fractionation of tissues revealed that the greatest specific activity and total activity, in all cases, was associated with the 105,000 g pellet. Significant activity was also detected in the 750 and 10,000 g pellets but not in the 105,000 g supernatant. Radioimmunoassay of endogenous estradiol-17 beta (estradiol) in chorion extracts revealed a 6.3-fold increase in the hormone from mid to late gestation. Estradiol levels in endometrium and myometrium did not appear to change during this time. It was concluded that increased estrogen sulfatase activity in guinea pig chorion in late gestation occurs along with elevated levels of the hormone estradiol which may be important for parturition in this species.     

In vitro steroid secretion by intact bovine ovarian follicles in a superfusion system

A superfusion system for single intact follicles is described. Bovine follicles were superfused for 5 to 10 hours. Progesterone and testosterone secretion was stable after an adaptation period of 150 min. A 5-minute stimulation with hCG resulted in a 2-fold increase in progesterone secretion. Superfusion for 450 minutes of 3 small follicles of similar size and in the same ovary next to each other showed different secretion patterns of progesterone and testosterone. Superfusion for 150 minutes of follicles obtained from 2 pairs of ovaries showed that follicular size is not a sufficient criterion to predict the progesterone and testosterone secretion pattern. Closeness of follicles (big or small) to a corpus luteum did not impair their ability to secrete progesterone and testosterone. It is concluded that a superfusion system as has been described in this report is an effective and desirable method to study the physiology and pathophysiology of single intact follicles.     

Factors affecting the tissues composition of pork belly

Bellies derived from the commercial population of pig carcasses are diverse in terms of tissue composition. Knowledge of the factors influencing it and the expected results, permits quick and easy evaluation of raw material. The study was designed to determine the factors affecting the tissues composition of pork bellies and to estimate their lean meat content. The research population (n=140 pig carcasses) was divided into groups according to sex (gilts, barrows), half-carcass mass (<40, 40 to 43.9, 44 to 46.9, ⩾47 kg) and lean meat content class: S (⩾60%), E (55% to 60%), U (50% to 55%), R (<50%). Bellies were subjected to a detailed dissection. Half-carcass mass affected the levels of all the analysed parameters. The only exception was the mass of the fat with the skin in the 40 to 43.9 kg group, for which the value did not differ statistically between the two groups <40 and 44 to 46.9 kg. Decrease in lean meat content affected the growth of the fat and skin mass in a linear way. No differences were observed between class S and E in terms of belly muscle mass. A 0.37% higher share of belly in the half-carcass was found for barrows (P<0.001), although bellies issued from barrows were characterized by a higher proportion of fat with skin compared with gilts (P=0.02). Interactions were observed between sex and half-carcass mass, so the sex of heavy half-carcasses becomes an important determinant for conditioning the muscle content. Equations were calculated and allow a fast and highly accurate determination of the lean meat content in bellies, suggesting they may be used directly in the production line.     

Factors affecting the glycosphingolipid composition of murine tissues

The effects of genetic strain, sex, age, and pathological state on the distribution and concentration of glycosphingolipids in mouse kidneys and livers were studied. The concentrations of glycosphingolipids and phospholipids in the kidneys and livers of different strains were compared. The major glycosphingolipid in the kidneys of male and female BALB/c, C3H/He, C57/BL, A, and C57xA (F(1) hybrid) mice was a sulfatide, monoglycosyl-(3-sulfate) ceramide; monoglycosyl ceramide was the major component in livers. The kidneys of males of all strains contained significant amounts of diglycosyl ceramide, but those of females contained, at most, only traces. Glycosphingolipid concentration in the male kidneys of C57/BL and C57xA (F(1) hybrid) was much higher than in the female and was also much higher than in the male kidneys of C3H/He, BALB/c, and A strains. The kidneys of "old" (36 wk) male and female C3H/He mice contained much higher proportions of monoglycosyl ceramide than 10-12-wk-old adults. The distributions of glycosphingolipids in kidneys of female C3H/He mice with BP8 ascites tumors and male C57xA (F(1) hybrid) mice with EL4 ascites tumors differed from those in the normal mice. An unknown lipid, present in all glycosphingolipid extracts from kidney and liver, was tentatively identified as cholesterol sulfate.