Biomedical subjects

F W Bazer

Publications and source records attributed to F W Bazer.

At least 19 recordsLinked to original sources

PubMed1998

Maternal dietary protein deficiency decreases amino acid concentrations in fetal plasma and allantoic fluid of pigs.

This study was conducted to test the hypothesis that maternal dietary protein deficiency decreases amino acid availability to the fetus, thereby contributing to retarded fetal growth. Primiparous gilts selected genetically for low or high plasma total cholesterol concentrations (low line and high line, respectively) were mated, and then fed 1.8 kg/d of isocaloric diets containing 13% or 0.5% crude protein. At d 40 or 60 of gestation, they were hysterectomized, and maternal and fetal blood samples as well as amniotic and allantoic fluids were obtained for analyses of amino acids, ammonia and urea. Dietary protein restriction decreased (P < 0.05) the following: 1) maternal plasma concentrations of urea at d 40 and 60 of gestation; 2) fetal plasma concentrations of alanine, arginine, branched-chain amino acids (BCAA), glutamine, glycine, lysine, ornithine, proline, taurine, threonine and urea at d 60 of gestation; 3) amniotic and allantoic fluid concentrations of urea at d 40 and 60 of gestation; and 4) allantoic fluid concentrations of alanine, arginine, BCAA, citrulline, cystine, glycine, histidine, methionine, proline, serine, taurine, threonine and tyrosine at d 40 of gestation, in gilts of both genetic lines. At d 60 of gestation, protein deficiency decreased (P < 0.05) allantoic fluid concentrations of arginine, cystine, glycine, taurine and tyrosine in low line gilts and of cystine, glutamine, ornithine, serine, taurine and tyrosine in high line gilts. Low line and high line gilts also differed remarkably in allantoic fluid concentrations of arginine, glutamine, ornithine and ammonia at d 40 and 60 of gestation. Our results suggest the following: 1) protein-deficient gilts maintain maternal plasma concentrations of amino acids by mobilizing maternal protein stores and decreasing oxidation of amino acids during the first half of gestation; 2) protein deficiency may impair placental transport of amino acids from the maternal to the fetal blood; and 3) low line and high line gilts differ in fetal amino acid metabolism. Decreases in concentrations of the essential and nonessential amino acids in the fetus may be a mechanism whereby maternal dietary protein restriction results in fetal growth retardation.

Allantois↗

PubMed1998

Mechanisms regulating prostaglandin F2 alpha secretion from the bovine endometrium.

The mechanism that regulates luteolytic PGF2 alpha secretion as stimulated by oxytocin is thought to involve induction of the inositol (1,4,5)-trisphosphate-diacylglycerol second messenger system, which mobilizes intracellular calcium and activates protein kinase C. In Experiment 1, endometrial explants taken from heifers on d 18.5 to 19.5 postestrus had increased PGF2 alpha secretion after treatment with 1 microM calcium ionophore A23187 to increase intracellular calcium, 100 nM phorbol 12-myristate 13-acetate to activate protein kinase C, and 100 nM oxytocin. The stimulatory effects of oxytocin and calcium ionophore A23187 plus phorbol 12-myristate 13-acetate did not differ from each other. In Experiment 2, endometrial explants taken from cows on d 18.5 to 19.5 postestrus had increased PGF2 alpha secretion after treatment with 0.2 and 2 microM thapsigargin to mobilize intracellular calcium that was sensitive to inositol (1,4,5)-trisphosphate. Secretion of PGF2 alpha was also increased by 100 nM oxytocin and was influenced by the interaction of thapsigargin and oxytocin such that 100 nM oxytocin did not further increase the secretion of PGF2 alpha in the presence of 2 microM thapsigargin. In Experiment 3, 100 nM oxytocin stimulated greater production of inositol trisphosphate and total inositol phosphates in the endometrium of cyclic cows than in the endometrium of pregnant cows on d 16.5 to 17.0 postestrus, although luteolysis was not yet initiated in the cyclic cows. These results are consistent with the hypothesis that the activation of the inositol (1,4,5)-trisphosphate-diacylglycerol second messenger system by oxytocin is involved in the stimulation of PGF2 alpha secretion from the endometrium during late diestrus in cows.

Animals↗

PubMed1997

Potent anti-feline immunodeficiency virus and anti-human immunodeficiency virus effect of IFN-tau.

Ovine IFN-tau is a newly described protein related to IFN-alpha that is responsible for maternal recognition of pregnancy in sheep. It has been shown to exhibit potent antiviral and antiproliferative activity. To determine its antiviral activity against feline immunodeficiency virus (FIV) and HIV, the activity of the RNA-dependent DNA polymerase, reverse transcriptase, was assayed in FIV- and HIV-infected feline and human PBL treated with IFN-tau. Significant dose-dependent inhibition of reverse transcriptase activity by IFN-tau was detected by day 6 of culture and was maintained through the peak of virus replication. In addition, production of the FIV core protein, p25, was blocked by IFN-tau. Both the amino- and carboxyl-terminal regions of IFN-tau, as identified by synthetic peptides, appear to be involved in its antiretroviral activity. Comparison of the anti-HIV activities of IFN-tau and recombinant human IFN-alpha2 (rHuIFN-alpha2) indicated that while rHuIFN-alpha2 was toxic to cells at 10,000 U/ml, IFN-tau antiretroviral activity was not associated with a decrease in either cell viability or immunologic reactivity. Thus, IFN-tau displayed potent anti-FIV and anti-HIV activity without the cytotoxicity associated with high concentrations of rHuIFN-alpha2.

Animals↗

PubMed1997

Mucosal morphogenesis and cytodifferentiation in fetal porcine small intestine.

BACKGROUND: Development of the small intestine is essential for proper nutrition of the fetus and the neonate. This investigation examines the morphogenesis and cytodifferentiation of developing fetal porcine small intestinal mucosa. METHODS: Fetuses were collected from gilts after hysterectomy. Small intestinal segments were removed and processed for light and electron microscopy. RESULTS: Fetal porcine small intestine developed from a simple tube of stratified epithelium to a tube containing villus and intervillus regions of simple columnar epithelium. This development occurred in a proximal to distal direction. By Day 40 of gestation, cytodifferentiation was evident with the presence of goblet cells and enteroendocrine cells in the duodenum. As development progressed, microvilli lengthened and components of the apical endocytic complex (AEC) were observed. By Day 110 of gestation, tubular and vesicular components of the AEC were confined to the jejunum, whereas large lysosomal vacuoles were observed in the distal jejunum. Duodenal epithelium at Day 110 was similar to postnatal epithelium. CONCLUSIONS: The pattern of fetal porcine small intestinal development is similar to that reported for fetal human small intestine. Villus development and cytodifferentiation occur at similar relative times in gestation when compared to the human. These observations support the use of the fetal pig as a model for investigations of human small intestinal development.

Animals↗

PubMed1997

Myelosuppression in the pig (Sus scrofa): uteroferrin reduces the myelosuppressive effects of 5-fluorouracil in young pigs.

The present study investigated the ability of uteroferrin to modulate the myelosuppressive effects of 5-fluorouracil (5-FU) in young pigs (Sus scrofa). Pigs (28-35 days of age; n = 6 per treatment) were infused with equal amounts of 5-FU on days 0 and 1 of the experimental period (37.5 mg/kg cumulative dose). Uteroferrin (100 micrograms/kg in 0.9% NaCl) or control (equivalent volume of 0.9% NaCl) was administered to pigs as intramuscular injections twice daily (08:00 and 20:00 hr) on days 1 through 21. Peripheral blood cell number, composition and progenitor cells were determined over 28 days. Treatment of pigs with 5-FU resulted in a rapid dose-dependent (P < 0.05) leukocytopenia. Concurrent treatment of pigs with uteroferrin reduced (P < 0.05) the rate of 5-FU-induced leukocytopenia (44 vs 77 +/- 7% decline from baseline on day 3) and enhanced (P < 0.05), the recovery from 5-FU on days 10 and 12 postinfusion. The positive effect of uteroferrin on leukocytes resulted primarily from a protection and/or enhanced recovery of neutrophils and monocytes. In addition, uteroferrin attenuated (P < 0.05) the suppression of red blood cell numbers after 5-FU administration (6.9 vs 6.1 +/- 0.2 x 10(6) cells/microliter on day 3), an affect reflected in increased hematocrit and hemoglobin concentrations. The effects of uteroferrin appeared to result from enhancement of the proliferation and/or differentiation of primitive pluripotent stem cells resistant to 5-FU, as concurrent treatment of pigs with uteroferrin resulted in a protection and/or enhanced recovery (P < 0.05) of CFU-GEMM, CFU-GM and BFU-E progenitor cells in the peripheral blood. These results are the first to demonstrate that uteroferrin can reduce the myelosuppressive effects of 5-FU in the pig and suggest that uteroferrin has hematopoietic growth factor activity in vivo.

Acid Phosphatase↗

PubMed1997

Uteroferrin and recombinant bovine GM-CSF modulate the myelosuppressive effects of 5-fluorouracil in young female pigs (Sus scrofa).

The present study investigated the ability of uteroferrin and recombinant bovine granulocyte monocyte/macrophage-colony stimulating factor (rbGM-CSF) to modulate the myelosuppressive effects of 5-fluorouracil (5-FU) in young female pigs (Sus scrofa). Pigs (N = 3/treatment) were infused with 5-FU (32.5 mg/kg) on days 0 and 1 of the experimental period. Uteroferrin (100 micrograms/kg in 0.9% NaCl), rbGM-CSF (10 micrograms/kg in 0.9% NaCl), uteroferrin + rbGM-CSF (as above) or control (0.9% NaCl) were administered as intramuscular injections twice daily (0800 and 2000 hr). Peripheral blood cell number, composition, and progenitor cells were determined over 28 days. Treatment of pigs with 5-FU resulted in a rapid leukocytopenia and thrombocytopenia (nadirs on days 5 and 7, respectively) and a modest decrease (P < 0.05) in red blood cell (RBC) number (nadir on day 14). Although nor affecting RBC and thrombocytes, treatment of pigs with uteroferrin had an initial protective effect (P < 0.05) on the 5-FU-induced leukocytopenia (63 and 64 vs 48 and 39 +/- 6% of baseline on days 3 and 5, respectively). In contrast, rbGM-CSF enhanced (P < 0.05) the rate of the leukocytopenia and had only minor effects on thrombocyte numbers relative to controls. These effects appeared to be additive, as pigs treated with uteroferrin + rbGM-CSF had a reduced rate of leukocytopenia compared to pigs treated with rbGM-CSF alone. Uteroferrin + rbGM-CSF also attenuated (P < 0.05) the suppression and enhanced (P < 0.05) recovery of RBC and thrombocyte numbers following 5-FU treatment. In control pigs, a modest rebound leukocytosis (122 +/- 6% of baseline) and thrombocytosis (141 +/- 9% of baseline) was evident. Uteroferrin enhanced (P < 0.05) the rebound leukocytosis (135 +/- 6% of baseline), but attenuated (P < 0.05) the thrombocytosis. In contrast, rbGM-CSF enhanced (P < 0.05) the duration of the leukocytosis during the recovery phase, an effect augmented by the combination of uteroferrin + rbGM-CSF. In addition, treatment with uteroferrin + rbGM-CSF resulted in a sustained thrombocytosis (days 12 to 21). As indicated by changes in CFU-GM, BFU-E, and CFU-GEMM progenitor cells in peripheral blood, the effects of uteroferrin and rbGM-CSF appeared to reflect their ability to enhance the proliferation and/or differentiation of both similar and distinct hematopoietic progenitor cells.

Acid Phosphatase↗

PubMed1997

The effect of uteroferrin and recombinant GM-CSF on hematopoietic parameters in normal female pigs (Sus scrofa).

The present study investigated the effect of uteroferrin and recombinant bovine granulocyte-monocyte/macrophage colony stimulating factor (rbGM-CSF) on hematopoiesis in young female pigs. Uteroferrin (100 micrograms/kg in 0.9% NaCl), rbGM-CSF (10 micrograms/kg in 0.9% NaCl), uteroferrin + rbGM-CSF (as above), or control (0.9% NaCl) were administered as intramuscular injections twice daily (0800 and 2000 hr). Peripheral blood cell number, composition, and progenitor cells were determined over 28 days. Uteroferrin had minimal effects on white blood cell (WBC) number, while rbGM-CSF caused both a rapid (days 2-7; maximum 122 +/- 8% of baseline) and late (days 16-28; maximum 133 +/- 8% of baseline) increase in WBC. Combination treatment with uteroferrin + rbGM-CSF abolished the initial increase in WBC number,but resulted in a prolonged increase in WBC number (days 14-28) relative to control. The rbGM-CSF-induced increase in WBC number resulted from rapid increases (P < 0.05) in monocytes and neutrophils. The addition of uteroferrin + rbGM-CSF enhanced (P < 0.05) the initial increase in the monocyte population and augmented the neutrophilia. In addition, uteroferrin + rbGM-CSF resulted in a dramatic eosinophilia (days 2-28), which was not detected in either the uteroferrin or rbGM-CSF treatments. Although not substantially affected by uteroferrin alone, rbGM-CSF caused an increase (P < 0.05) in thrombocyte numbers from days 1 through 9 (maximum 133 +/- 11% of baseline), an effect augmented by cotreatment with uteroferrin. The ability of these cytokines to modulate blood cell number and composition appeared to result from their effects on hematopoietic progenitor cells. Treatment of pigs with uteroferrin increased (P < 0.05) CFU-GEMM, CFU-GM, and BFU-E progenitor cells in peripheral blood, while rbGM-CSF caused increases (P < 0.05) relative to control in CFU-GM and CFU-GEMM. These effects were additive, as uteroferrin + GM-CSF augmented the increases in CFU-GM, BFU-E, and CFU-GEMM. Collectively, these results indicate that uteroferrin and rbGM-CSF can modulate hematopoiesis in young pigs. These effects were both additive and, in the case of neutrophils and eosinophils, synergistic. Hence, the mechanism(s) by which uteroferrin and rbGM-CSF modulate hematopoiesis appear to be different.

Acid Phosphatase↗

PubMed1997

Lack of effect of granulocyte-macrophage colony-stimulating factor on secretion of interferon-tau, other proteins, and prostaglandin E2 by the bovine and ovine conceptus.

Three experiments tested the effects of recombinant bovine granulocyte-macrophage colony-stimulating factor (rbGM-CSF) on the preimplantation bovine and ovine conceptus. There was no effect of rbGM-CSF on the secretion of total radiolabeled protein in conditioned medium, immunoreactive interferon-tau (IFN tau), antiviral activity, or prostaglandin E2 from Day 16-18 bovine conceptuses cultured for 24 hr with, [3H]leucine and +/- 10 ng/ml rbGM-CSF. Similarly, there was no effect of 1 ng/ml rbGM-CSF on the secretion of total radiolabeled protein. IFN tau, or antiviral activity from Day 17 ovine conceptuses. There was also no beneficial effect of 1 or 10 ng/ml rbGM-CSF on the presence of immunoreactive IFN tau in conditioned medium from in vitro-produced bovine blastocysts at Day 7-8 after fertilization. Results indicate that IFN tau secretion from bovine and ovine conceptuses are unresponsive to rbGM-CSF at the concentrations tested.

Animals↗

PubMed1997

Recombinant ovine and bovine interferons tau regulate prostaglandin production and oxytocin response in cultured bovine endometrial cells.

At the time of recognition of pregnancy, the bovine conceptus (embryo and associated membranes) must produce a signal that will prevent luteolysis otherwise induced by pulsatile release of prostaglandin (PG) F2alpha from the uterus in response to oxytocin (OT). In ruminants, trophoblastic interferon tau (IFN-tau) released by the conceptus appears to be the most likely candidate to trigger the establishment of pregnancy. We have compared the effect of recombinant (r) ovine (o) and bovine (b) IFN-tau on PG production, using a fully characterized model of cultured endometrial cells. In uterine epithelial cells, the production of PGF2alpha was stimulated 7.1-fold (p < 0.0001) and that of PGE2 89.0-fold (p < 0.0001) by rbIFN-tau, and 3.6-fold and 29-fold, respectively, by roIFN-tau. The stimulation resulted in a net increase in the PGE2:PGF2alpha ratio of 7.7 with rbIFN-tau and 5.1 with roIFN-tau at the optimal concentration of 1 microg/ml (p < 0.0001). By contrast, addition of OT (100 nM) alone resulted in a decrease of the PGE2:PGF2alpha ratio. The level of stimulation of PGE2 by IFN-tau was reduced in the presence of OT, showing that there was some interaction between OT and IFN-tau at the cellular level in the regulation of PG production. In uterine stromal cells, roIFN-tau and rbIFN-tau stimulated PGE2 and PGF2alpha production 12-fold (p < 0.0001). The ratio of PGE2:PGF2alpha was not affected in a dose-dependent manner, but was increased (p < 0.001) at a single dose of rbIFN-tau (0.001 microg/ml) and roIFN-tau (0.01 microg/ml). The results indicate that 1) bovine endometrial cells are more responsive to rbIFN-tau than to roIFN-tau, 2) rIFN-tau regulates PGs by stimulating PGE2 preferentially, and 3) rIFN-tau transforms the response to OT from stimulation of PGF2alpha to stimulation of PGE2.

Animals↗

PubMed1997

Spatial and temporal analyses of integrin and Muc-1 expression in porcine uterine epithelium and trophectoderm in vitro.

The expression of the glycoproteins Muc-1 and integrin subunits (alpha1, alpha3, alpha4, alpha5, alpha(v), beta1, and beta3), all of which may be involved in the control of uterine receptivity, was examined in cultured porcine uterine epithelial (pUE) cells. Integrin subunit expression was also determined in trophoblastic vesicles generated from Day 12 and Day 15 pig conceptuses. Immunocytochemistry was performed on pUE cells cultured on several substrates (glass, serum, fibronectin, or Matrigel) in DME/F12 medium with 5% charcoal/dextran-stripped fetal bovine serum either containing no steroids or supplemented with estrogen, progesterone, or estrogen plus progesterone, or in medium with 5% complete fetal bovine serum. In addition, pUE cells were grown on Matrigel-coated Millicell filter inserts to facilitate development of structurally and functionally distinct apical and basal domains (i.e., polarized pUE cells) and were maintained in the same medium with supplements. The major findings in this investigation are that 1) Muc-1 and alpha1, alpha3, alpha4, alpha5, alpha(v), beta1, and beta3 integrin subunits are expressed on pUE cells in vitro; 2) Muc-1 and several integrin subunits are modulated by steroid hormones if culture conditions are provided that permit development of cell polarity; 3) the expression and steroid-induced modulation of Muc-1 and several integrin subunits in polarized pUE cells are similar to those detected in vivo; and 4) the expression of alpha1, alpha3, alpha4, alpha5, alpha(v), beta1, and beta3 integrin subunits by trophoblastic vesicles is identical to their expression on trophectoderm of intact conceptuses in vivo. These results suggest that the properties of polarized pUE cells and trophoblastic vesicles are comparable to those of their counterparts in vivo and are, therefore, useful as in vitro model systems to study conceptus-endometrial interactions during the periimplantation period and the control of uterine receptivity to implantation.

Animals↗

PubMed1997

Immunolocalization of acidic and basic fibroblast growth factors in porcine uterine and conceptus tissues.

Immunohistochemical techniques were used to localize acidic and basic fibroblast growth factor (FGF) polypeptides in porcine uterine and conceptus tissues collected on Days 10 through 14 of gestation, which is the peri-implantation period, and in uterine tissues collected on the same days of the estrous cycle. Our results demonstrate differential expression of acidic and basic FGF (aFGF, bFGF) in porcine uterine and conceptus tissues. Localization of these peptides in the uterus of cycling and pregnant pigs was different from that reported for other species, suggesting species-specific roles for FGFs in early pregnancy. Increases in both cytoplasmic and nuclear bFGF immunostaining were detected in uterine luminal and glandular epithelial cells from Days 10 to 14 of gestation but not in the uterine epithelium of cycling pigs. Acidic FGF immunostaining was not detected in luminal or glandular epithelium of either cycling or pregnant uterine tissue; however, differential stromal staining was observed. Uterine tissues collected from pigs on Days 10-14 of the estrous cycle had diffuse aFGF immunostaining throughout the stroma. During early pregnancy, however, intense aFGF immunostaining was concentrated around the glandular epithelial and below the luminal epithelial cells as gestation progressed. Basic FGF, but not aFGF, was detected in porcine conceptuses collected during the peri-implantation period (Days 10-14 of gestation). Although both acidic and basic FGF belong to the same family of proteins, results of the present study indicate that they likely play different roles in uterine function and conceptus development in pigs.

Animals↗

PubMed1997

Effects of exogenous recombinant ovine interferon tau on circulating concentrations of progesterone, cortisol, luteinizing hormone, and antiviral activity; interestrous interval; rectal temperature; and uterine response to oxytocin in cyclic ewes.

Interferon tau (IFN tau) is the conceptus-produced antiluteolytic signal in ruminants. Three experiments examined the effects of s.c. administration of recombinant ovine (ro)IFN tau on interestrous interval (IEI), oxytocin (OT)-induced uterine prostaglandin F2alpha metabolite (PGFM) production, rectal temperature (RT), respiration rate (RR), and plasma concentrations of progesterone, cortisol, LH, and antiviral activity (AVA) in plasma and uterine flushings. In experiment I, 20 ewes were treated s.c. with either 0, 1, 2, or 4 mg/day roIFN tau (0.7 x 10(8) U/mg; 5 ewes/dosage) from Days 11 to 15 of the estrous cycle (estrus = Day 0) and were challenged with OT (30 IU) on Day 15. Jugular blood samples were collected at -10, 0, 10, 20, 30, 40, 50, and 60 min relative to the OT challenge and assayed for PGFM. Recombinant oIFN tau increased IEI (16.7, 18.7, and 22.6 +/- 0.6 days for 0, 2, and 4 mg roIFN tau, respectively, p < 0.01). Recombinant oIFN tau did not affect peak PGFM response to OT (2309 +/- 172 pg/ml; p > 0.1). However, the 4 mg/day dosage delayed the time to peak PGFM (32.4 vs. 47.5 +/- 3.4 min; p < 0.01, 0 vs. 4 mg) and resulted in approximately 200% higher concentrations of PGFM at 60 min post-OT (0 vs. 4 mg/day, p < 0.07). Experiment II was similar to experiment I, except that only the 0- and 4-mg/day dosages of roIFN tau were administered. Ewes were hysterectomized on Day 16, and assay of uterine flushes detected no AVA from ewes treated with either 0 or 4 mg/day roIFN tau. In experiment III, 20 ewes were treated s.c. with either 0, 2, 4, or 6 mg roIFN tau on Day 12. Blood samples, RT, and RR were obtained at frequent intervals for 24 h, and plasma was assayed for progesterone, cortisol, LH, and AVA. Plasma AVA, which increased in a dose-dependent manner, was detectable within 60 min and remained elevated at 24 h compared to control values. RT (elevated 0.5-1.0 degrees C), RR, and cortisol increased in response to all dosages of roIFN tau, with peak values occurring 150-180 min postinjection. For all dosages of roIFN tau, plasma progesterone declined from 120 to 360 min posttreatment and then returned to pretreatment values by 24 h (p < 0.01) as compared to controls. Overall, exogenous roIFN tau altered uterine PGFM response to OT from a pulse to a gradual and sustained elevation and extended IEI with only a transient decline in progesterone and mild hyperthermia, effects that are not expected to compromise pregnancy.

Animals↗

PubMed1997

Interferon tau: a novel pregnancy recognition signal.

PROBLEM: Trophectoderm of ruminant conceptuses (embryo and associated membranes) secretes tau interferons (IFNtau) as the pregnancy recognition signal. How does it act? METHOD: Review of current data. RESULTS: IFNtau acts on uterine epithelium to suppress transcription of the genes for estrogen receptor and oxytocin receptor. This blocks development of the uterine luteolytic mechanism and, therefore, release of luteolytic pulses of prostaglandin F2alpha, but it has no effect on expression of the progesterone receptor. Maintenance of progesterone secretion by the corpus luteum ensures establishment and maintenance of pregnancy. Secretion of IFNtau on days 12-15 for sheep and days 14-17 for cows and goats is essential for pregnancy recognition. CONCLUSION: We propose that IFNtau affects endometrial gene expression by activating the Jak/Stat pathway, which results in formation of the ISGF3alpha transcription factor complex. ISGF3alpha binds to interferon-stimulated response elements and activates transcription of interferon-responsive genes such as interferon regulatory factor-1 (IRF-1) which, in turn, activates expression of the negative-acting transcription factor IRF-2. Pregnancy (or intrauterine injection of roIFNtau) results in a transient increase in endometrial IRF-1 expression followed 36-48 hr later by a sustained increase in IRF-2. We propose that IRF-2, or an IFNtau-induced negative regulatory factor like IRF-2, suppresses expression of the estrogen receptor gene and directly or indirectly blocks expression of the gene for oxytocin receptor to abrogate the uterine luteolytic mechanism and ensure the establishment of pregnancy.

Animals↗

PubMed1996

Developmentally regulated expression of interleukin-1 beta by peri-implantation conceptuses in swine.

Interleukin-1 beta (IL-1 beta) is one of the critical inflammatory cytokines involved in many physiological processes. This study investigated the temporal expression of IL-1 beta in pig conceptuses. A human IL-1 beta cDNA probe and a anti-human IL-1 beta antibody were used to examine IL-1 beta gene and protein expression in pig conceptuses on days 11, 12, 13, 14, 15, 20, 30, 45, 60, 75, 90, 105 and 112 of pregnancy using Northern, Slot and Western blot analyses. A human cell line (A375.S2) was used to determine IL-1 activity in pig conceptuses. High levels of IL-1 beta mRNA were detected in days 11, 12 and 13 conceptuses. IL-1 beta protein was also detected in conceptuses on days 11, 12, 13, and 14, but not in conceptuses recovered after day 15 of pregnancy. IL-1 beta biological activity was demonstrated in days 11 and 12 conceptus homogenates, but not in homogenates of days 112 allantochorion. Low levels of IL-1 beta mRNA were detected by Northern blot analysis in Day 15 conceptuses, endometrium and myometrium only when poly(A+) RNA was used. The production of IL-1 beta by peri-implantation pig conceptuses was temporally associated with maternal recognition of pregnancy. The results suggest that conceptus IL-1 beta may be important for conceptus development and establishment of pregnancy.

Animals↗

PubMed1996

Transfer of heterologous immunoglobulin into the uterine lumen of pigs.

Transfer of circulating heterologous immunoglobulin G (IgG) into the uterine lumen of pigs has not been reported. The present study determined if ovine IgG (oIgG) could be transferred into the uterine lumen of pigs. Six gilts (nonparous female pigs) were injected i.v. with either immune sheep serum (25 or 50 ml) to porcine uteroferrin (Uf) or non-immune sheep serum (50 ml) on days 9, 11 and 13 of the estrous cycle. Serum was collected daily from days 9 to 15 and uterine flushings were collected at hysterectomy on day 15. An ELISA detecting oIgG was used to determine levels of oIgG in pig sera and uterine flushings. High oIgG levels in serum (ranging from 87 +/- 11 to 141 +/- 14 micrograms/ml) were maintained by injecting the gilts at 48 h intervals with ovine antiserum to porcine Uf. Serum concentrations of oIgG did not differ (P > 0.05) regardless of whether immune or non-immune sera or different doses of immune serum were injected. oIgG in uterine flushings (2 +/- 1 micrograms/uterine flushing) was detectable when the samples were concentrated 40-fold, but were lower (P < 0.01) than serum levels of oIgG (107 +/- 10 micrograms/ml). Results indicate that small amounts of circulating heterologous IgG can be transferred into the uterine lumen of pigs. However, passive immunization may not result in titers high enough to examine in vivo functions of proteins secreted into the uterine lumen of pigs.

Acid Phosphatase↗

PubMed1996

Effects of interferon-tau and progesterone on oestrogen-stimulated expression of receptors for oestrogen, progesterone and oxytocin in the endometrium of ovariectomized ewes.

The effects of recombinant ovine interferon-tau (IFN-tau) and progesterone on oestrogen-stimulated expression of endometrial receptors for oestrogen (ER), progesterone (PR) and oxytocin (OTR) were determined in ovariectomized ewes. Cyclic ewes (n = 16) were ovariectomized and fitted with uterine catheters on Day 4 of the oestrous cycle (Day O, oestrous) and assigned randomly in 2 x 2-factorial arrangement to receive daily intrauterine injections of either recombinant ovine IFN-tau (roIFN-tau; 2 x 10(7) anti-viral units) or control proteins from Day 11 to Day 15 and 50 mg progesterone from either Day 4 to Day 10 (E-P) or Day 4 to Day 15 (E+P). All ewes received 50 micrograms oestradiol-17 beta on Days 13, 14 and 15 and were hysterectomized on Day 16. In control ewes, endometrial ER mRNA, PR protein and OTR density were greater in E-P- than E+P- treated ewes. In E-P ewes, roIFN-tau decreased oestrogen-stimulated increases in ER and OTR, but not PR expression compared with control ewes. In E+P ewes, endometrial ER mRNA and protein, PR mRNA and protein, and OTR levels were lower in roIFN-tau-treated ewes than control ewes. Immunoreactive ER and PR were absent in the endometrial luminal and superficial glandular epithelium of roIFN-tau compared with control ewes, but were present in the deep glandular epithelium and stroma regardless of steroid or protein treatment. These results indicate that progesterone affects oestrogen-induced increases in endometrial ER, PR and OTR expression in the PR+ deep glandular epithelium and stroma, whereas IFN-tau suppresses oestrogen-induced increases ER, PR and OTR expression in the PR- luminal and superficial glandular epithelium. These combined actions of IFN-tau and progesterone to suppress oestrogen-induced increases in endometrial OTR formation would prevent pulsatile production of luteolytic prostaglandin F2 alpha by the endometrium during early pregnancy.

Animals↗

PubMed1996

Expression of oncofetal fibronectin in porcine conceptuses and uterus throughout gestation.

Oncofetal fibronectin is reportedly expressed specifically by trophoblast tissue and some tumours and speculated to mediate placental attachment. In the present study, a monoclonal antibody (FDC-6) to human oncofetal fibronectin was used to characterize temporal and spatial changes in the expression of oncofetal fibronectin at the fetal-maternal interface of pigs. Conceptus and uterine tissues were collected from gilts throughout normal pregnancy and processed for immunohistochemical characterization. Results indicated that oncofetal fibronectin was constitutively expressed by both porcine conceptus and uterus throughout gestation. The most abundant staining for oncofetal fibronectin was found in the allantochorion and detectable levels of oncofetal fibronectin were also detected in luminal and glandular epithelial cells in the uterus. During the second-half of pregnancy, oncofetal fibronectin was also detected in fibroblast-like cells in the uterine stroma, but not in the stroma of the allantochorion. Owing to the non-invasive nature of the porcine placenta, the abundant expression of oncofetal fibronectin by the trophoblast and uterus may influence attachment between chorion and endometrium during pregnancy. Since attachment is the first step in implantation and placentation in all mammalian species, the pig may represent an excellent animal model to study interactions between trophoblast and endometrium mediated by oncofetal fibronectin.

Animals↗

PubMed1996

High yield expression and secretion of the ovine pregnancy recognition hormone interferon-tau by Pichia pastoris.

The early conceptus (embryo and associated membranes) of domestic ruminats signals its presence to the maternal uterus through production of interferon-tau (IFN-tau). Production of IFN-tau ensures continued production of progesterone, the hormone of pregnancy, by the ovarian corpus luteum. This paper reports the high-level expression and efficient secretion of biologically active recombinant ovine IFN-tau (rOvIFN-tau) by Pichia pastoris. The developed method produces more than 80% pure recombinant ovine IFN-tau, obviating the need for further purification for many purposes. Initial fermentation studies produced IFN-tau at 280 mg/liter and demonstrate the potential of this system for large-scale production of IFN-tau.

Animals↗