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1.
J Dairy Sci ; 100(11): 9402-9417, 2017 Nov.
Article in English | MEDLINE | ID: mdl-28918138

ABSTRACT

Inadequate feed consumption reduces intestinal barrier function in both ruminants and monogastrics. Objectives were to characterize how progressive feed restriction (FR) affects inflammation, metabolism, and intestinal morphology, and to investigate if glucagon-like peptide 2 (GLP2) administration influences the aforementioned responses. Twenty-eight Holstein cows (157 ± 9 d in milk) were enrolled in 2 experimental periods. Period 1 [5 d of ad libitum (AL) feed intake] served as baseline for period 2 (5 d), during which cows received 1 of 6 treatments: (1) 100% of AL feed intake (AL100; n = 3), (2) 80% of AL feed intake (n = 5), (3) 60% of AL feed intake (n = 5), (4) 40% of AL feed intake (AL40; n = 5), (5) 40% of AL feed intake + GLP2 administration (AL40G; 75 µg/kg of BW s.c. 2×/d; n = 5), or (6) 20% of AL feed intake (n = 5). As the magnitude of FR increased, body weight and milk yield decreased linearly. Blood urea nitrogen and insulin decreased, whereas nonesterified fatty acids and liver triglyceride content increased linearly with progressive FR. Circulating endotoxin, lipopolysaccharide binding protein, haptoglobin, serum amyloid A, and lymphocytes increased or tended to increase linearly with advancing FR. Circulating haptoglobin decreased (76%) and serum amyloid A tended to decrease (57%) in AL40G relative to AL40 cows. Cows in AL100, AL40, and AL40G treatments were euthanized to evaluate intestinal histology. Jejunum villus width, crypt depth, and goblet cell area, as well as ileum villus height, crypt depth, and goblet cell area, were reduced (36, 14, 52, 22, 28, and 25%, respectively) in AL40 cows compared with AL100 controls. Ileum cellular proliferation tended to be decreased (14%) in AL40 versus AL100 cows. Relative to AL40, AL40G cows had improved jejunum and ileum morphology, including increased villus height (46 and 51%), villus height to crypt depth ratio (38 and 35%), mucosal surface area (30 and 27%), cellular proliferation (43 and 36%), and goblet cell area (59 and 41%). Colon goblet cell area was also increased (48%) in AL40G relative to AL40 cows. In summary, progressive FR increased circulating markers of inflammation, which we speculate is due to increased intestinal permeability as demonstrated by changes in intestinal architecture. Furthermore, GLP2 improved intestinal morphology and ameliorated circulating markers of inflammation. Consequently, FR is a viable model to study consequences of intestinal barrier dysfunction and administering GLP2 appears to be an effective mitigation strategy to improve gut health.


Subject(s)
Cattle/physiology , Food Deprivation , Glucagon-Like Peptide 2/pharmacology , Inflammation/veterinary , Intestines/drug effects , Animals , Biomarkers/blood , Body Weight , Cattle/blood , Diet/veterinary , Fatty Acids, Nonesterified/blood , Female , Inflammation/blood , Inflammation/metabolism , Intestinal Mucosa/drug effects , Intestines/physiology , Lactation , Milk
2.
Mucosal Immunol ; 9(6): 1571-1583, 2016 11.
Article in English | MEDLINE | ID: mdl-27007679

ABSTRACT

Depot-medroxyprogesterone acetate (DMPA) is a hormonal contraceptive especially popular in areas with high prevalence of HIV and other sexually transmitted infections (STI). Although observational studies identify DMPA as an important STI risk factor, mechanisms underlying this connection are undefined. Levonorgestrel (LNG) is another progestin used for hormonal contraception, but its effect on STI susceptibility is much less explored. Using a mouse model of genital herpes simplex virus type 2 (HSV-2) infection, we herein found that DMPA and LNG similarly reduced genital expression of the desmosomal cadherin desmoglein-1α (DSG1α), enhanced access of inflammatory cells to genital tissue by increasing mucosal epithelial permeability, and increased susceptibility to viral infection. Additional studies with uninfected mice revealed that DMPA-mediated increases in mucosal permeability promoted tissue inflammation by facilitating endogenous vaginal microbiota invasion. Conversely, concomitant treatment of mice with DMPA and intravaginal estrogen restored mucosal barrier function and prevented HSV-2 infection. Evaluating ectocervical biopsy tissue from women before and 1 month after initiating DMPA remarkably revealed that inflammation and barrier protection were altered by treatment identically to changes seen in progestin-treated mice. Together, our work reveals DMPA and LNG diminish the genital mucosal barrier; a first-line defense against all STI, but may offer foundation for new contraceptive strategies less compromising of barrier protection.


Subject(s)
Contraceptive Agents, Female/pharmacology , Disease Susceptibility , Herpes Genitalis/virology , Herpesvirus 2, Human/drug effects , Levonorgestrel/pharmacology , Medroxyprogesterone Acetate/pharmacology , Mucous Membrane/drug effects , Mucous Membrane/virology , Animals , Disease Models, Animal , Estrogens/pharmacology , Female , Mice , Mucous Membrane/metabolism , Permeability/drug effects , Vagina/drug effects , Vagina/immunology , Vagina/metabolism , Vagina/virology
3.
J Anim Sci ; 93(3): 1229-38, 2015 Mar.
Article in English | MEDLINE | ID: mdl-26020899

ABSTRACT

The effect of feeding ractopamine hydrochloride (RAC) on growth performance, carcass and pork quality, and blood acid-base and catecholamine responses to handling and transport in finishing pigs was evaluated using a randomized complete block design to compare 2 RAC levels (0 vs. 10 mg/kg). Crossbred pigs ( = 144) were housed in single-sex pens (barrow or gilt) of 3 with 24 pens/RAC level. The study was carried out for a 28-d period from 104.0 ± 5.99 to 136.7 ± 6.44 kg BW. At the end of the growth study, pigs were subjected to handling and transport procedures that involved an initial aggressive handling procedure (pigs moved 50 m with 8 shocks from an electric prod) followed by a 30-min transport on a standard livestock trailer at a floor space of 0.46 m/pig followed by a final gentle handling procedure (pigs moved 100 m using sort boards and slap paddles). A blood sample was taken and rectal temperature was measured 2 h before (baseline) and immediately after the final handling procedure (final). Barrows ( = 72) were harvested and carcass and pork quality were measured. Feeding RAC increased ( ≤ 0.05) ADG (19.6%), ADFI (4.2%), and G:F (14.8%). The increase in plasma epinephrine levels from baseline to final was greater ( ≤ 0.05) for pigs fed RAC; there was a trend ( ≤ 0.10) for pigs fed RAC to have greater final blood lactate and to show a greater change from baseline to final in blood bicarbonate, partial pressure of and total carbon dioxide, and oxygen saturation levels. However, there were no differences between treatments for changes from baseline to final in rectal temperature, blood pH and lactate, and plasma norepinephrine levels. The incidence of physical indicators of stress and of nonambulatory, noninjured pigs during the handling and transport procedures was similar for the 0 and 10 mg/kg RAC levels. Final farm BW was 4.1 kg heavier, carcass yield was 1.4 percentage units greater, and LM area was 5.18 cm greater for pigs fed RAC compared to the control ( ≤ 0.05). Minolta a* and b* values were lower ( ≤ 0.05) and ultimate pH (0.05 units) and Warner-Bratzler shear force (0.43 kg) were greater ( ≤ 0.05) for pigs fed 10 compared to 0 mg/kg RAC. These results confirm the substantial improvement from feeding 10 mg/kg RAC in growth performance and carcass yield and suggest relatively limited effects on pork quality and on responses to the handling and transport procedures used in this study.


Subject(s)
Meat/standards , Phenethylamines/pharmacology , Adrenergic beta-Agonists/pharmacology , Animals , Body Composition/drug effects , Female , Male , Swine/growth & development
4.
N Engl J Med ; 345(7): 487-93, 2001 Aug 16.
Article in English | MEDLINE | ID: mdl-11519502

ABSTRACT

BACKGROUND: Infection with Trichomonas vaginalis during pregnancy has been associated with preterm delivery. It is uncertain whether treatment of asymptomatic trichomoniasis in pregnant women reduces the occurrence of preterm delivery. METHODS: We screened pregnant women for trichomoniasis by culture of vaginal secretions. We randomly assigned 617 women with asymptomatic trichomoniasis who were 16 to 23 weeks pregnant to receive two 2-g doses of metronidazole (320 women) or placebo (297 women) 48 hours apart. We treated women again with the same two-dose regimen at 24 to 29 weeks of gestation. The primary outcome was delivery before 37 weeks of gestation. RESULTS: Between randomization and follow-up, trichomoniasis resolved in 249 of 269 women for whom follow-up cultures were available in the metronidazole group (92.6 percent) and 92 of 260 women with follow-up cultures in the placebo group (35.4 percent). Data on the time and characteristics of delivery were available for 315 women in the metronidazole group and 289 women in the placebo group. Delivery occurred before 37 weeks of gestation in 60 women in the metronidazole group (19.0 percent) and 31 women in the placebo group (10.7 percent) (relative risk, 1.8; 95 percent confidence interval, 1.2 to 2.7; P=0.004). The difference was attributable primarily to an increase in preterm delivery resulting from spontaneous preterm labor (10.2 percent vs. 3.5 percent; relative risk, 3.0; 95 percent confidence interval, 1.5 to 5.9). CONCLUSIONS: Treatment of pregnant women with asymptomatic trichomoniasis does not prevent preterm delivery. Routine screening and treatment of asymptomatic pregnant women for this condition cannot be recommended.


Subject(s)
Antitrichomonal Agents/therapeutic use , Metronidazole/therapeutic use , Obstetric Labor, Premature/prevention & control , Pregnancy Complications, Parasitic/drug therapy , Trichomonas Vaginitis/drug therapy , Adult , Animals , Female , Follow-Up Studies , Humans , Infant, Newborn , Infant, Premature , Pregnancy , Pregnancy Complications , Treatment Failure , Trichomonas vaginalis/isolation & purification , Vagina/parasitology
5.
Am J Obstet Gynecol ; 180(3 Pt 1): 578-80, 1999 Mar.
Article in English | MEDLINE | ID: mdl-10076131

ABSTRACT

OBJECTIVE: The purpose of this study was to determine whether digital examination introduces vaginal organisms into the cervix. STUDY DESIGN: Thirty-five women with reported ruptured membranes at >/=34 weeks' gestation underwent a sterile speculum examination and a standardized semiqualitative, semiquantitative endocervical culture before and immediately after digital cervical examination. RESULTS: Cultures taken before digital examination demonstrated a mean of 2.8 +/- 1.7 different types of organisms, whereas cultures taken after digital examination demonstrated a mean of 4.4 +/- 1.5 different types of organisms (P <.0001). Twenty-eight patients (80%) had heavier growth or a greater number of different organisms in the postexamination culture than in the pre-examination culture. The state of the fetal membranes (ruptured as opposed to intact) did not alter these relationships. CONCLUSION: An immediate effect of digital examination is the introduction of vaginal organisms into the cervical canal.


Subject(s)
Cervix Uteri/microbiology , Fetal Membranes, Premature Rupture/microbiology , Obstetrics , Physical Examination/adverse effects , Pregnancy Complications, Infectious/microbiology , Adolescent , Adult , Female , Humans , Obstetrics/methods , Pregnancy
6.
J Anim Sci ; 76(10): 2657-70, 1998 Oct.
Article in English | MEDLINE | ID: mdl-9814907

ABSTRACT

This experiment consisted of the following treatment-breed groups: 1) White crossbred gilts, 2) White crossbred gilts treated with progesterone (200 mg/d in corn oil given on d 2 and 3 after estrus), and 3) Chinese Meishan gilts. Pregnant and nonpregnant gilts (n=3 to 6) from each treatment-breed combination were assigned to be slaughtered on d 10, 11, 12, 13, and 15. At slaughter each uterine horn was flushed with 20 mL of minimal essential medium. Uterine flushings were assayed for total protein, acid phosphatase, uteroferrin, retinol-binding protein, and oxytocin. Uterine flush total protein was increased by progesterone treatment, was unaffected by pregnancy status, and was less in Meishans. Similar patterns were found for retinol binding protein and uteroferrin, except that uteroferrin was greater in pregnant than in nonpregnant gilts. Oxytocin was greater in pregnant than in nonpregnant gilts, was not influenced by progesterone treatment, and was similar in Meishan and in White crossbred gilts. These results indicate that the conceptus does not influence secretion of either total protein or retinol binding protein during pregnancy and that the onset of secretion of these uterine proteins may be controlled by progesterone. The presence of the conceptus is associated with increased uteroferrin and oxytocin production. The decreased secretion of uterine proteins in Meishan gilts may partially explain the slower embryonic development that has been reported for this breed.


Subject(s)
Breeding , Pregnancy, Animal/metabolism , Progesterone/pharmacology , Proteins/metabolism , Swine/metabolism , Uterus/metabolism , Acid Phosphatase/analysis , Acid Phosphatase/genetics , Acid Phosphatase/metabolism , Analysis of Variance , Animals , Blotting, Northern , Estrogens/analysis , Estrogens/genetics , Estrogens/metabolism , Female , Isoenzymes , Least-Squares Analysis , Metalloproteins/analysis , Metalloproteins/genetics , Metalloproteins/metabolism , Oxytocin/analysis , Oxytocin/genetics , Oxytocin/metabolism , Pregnancy , Proteins/analysis , Proteins/genetics , RNA, Messenger/analysis , Radioimmunoassay/veterinary , Retinol-Binding Proteins/analysis , Retinol-Binding Proteins/genetics , Retinol-Binding Proteins/metabolism , Tartrate-Resistant Acid Phosphatase , Uterus/drug effects
7.
Anim Reprod Sci ; 47(1-2): 123-35, 1997 May.
Article in English | MEDLINE | ID: mdl-9233512

ABSTRACT

Objectives were to examine the effects of a single dose (4 mg) of estradiol-17 beta (E2) on blastocyst development around the period of elongation. Proestrus gilts were induced to ovulate with 750 IU of hCG and were mated before ovulation (normal mating, 24 to 32 h post-hCG) or after ovulation had begun (delayed mating, 43 h post-hCG). This difference in time of mating has been demonstrated to result in approximately a 7-h difference in time of blastocyst elongation. Normally and delay-mated gilts were ovariohysterectomized at 278 h post-hCG or injected with E2 or vehicle (corn oil) at 278 h and then ovariohysterectomized at 290 h post-hCG (five or six gilts per group). Blastocyst size was measured and concentrations of E2, retinol, uteroferrin, insulin-like growth factor-I (IGF-I), uterine plasmin/trypsin inhibitor (UPTI) and protein in uterine flushings were quantified. Blastocyst size and components of uterine flushings did not differ (P > 0.05) between normally and delay-mated gilts at 278 h post-hCG. However, at 290 h post-hCG, normally mated gilts had larger (P < 0.01) blastocysts (small spheres to filamentous) and their flushings tended to contain less (P < 0.07) amounts of retinol than those of delay-mated gilts whose blastocysts ranged from small spheres to ovoidals. Normally mated gilts receiving E2 at 278 h had smaller (P < 0.01) blastocysts and less (P < 0.05) amounts of retinol at 290 h post-hCG than gilts receiving vehicle. Conversely, delay-mated gilts treated with E2 or vehicle did not differ (P > 0.05) in blastocyst size and amounts of components of uterine flushings at 290 h post-hCG. Normally mated gilts treated with vehicle had litters in the process of elongating at 290 h post-hCG. Mean blastocyst size (P < 0.001) and amounts of components of uterine flushings (except for IGF-I) in these gilts were greater (P < 0.05, UPTI = 0.06) than in normally mated gilts at 278 h post-hCG, whose blastocysts were spherical. Among gilts not treated with E2 (278 h and 290 h pooled), mean blastocyst size was positively correlated (P < 0.05) with amounts of retinol, E2, uteroferrin and total protein. Results indicated that a single dose of E2 given before elongation altered blastocyst development depending on how close blastocysts were to onset of elongation at the time of E2 treatment.


Subject(s)
Blastocyst/drug effects , Embryonic and Fetal Development/drug effects , Estradiol/pharmacology , Swine/embryology , Acid Phosphatase , Analysis of Variance , Animals , Chorionic Gonadotropin/pharmacology , Embryonic and Fetal Development/physiology , Estradiol/analysis , Female , Insulin-Like Growth Factor I/analysis , Isoenzymes , Male , Metalloproteins/analysis , Pregnancy , Swine/physiology , Tartrate-Resistant Acid Phosphatase , Time Factors , Uterus/chemistry , Vitamin A/analysis
8.
Mol Reprod Dev ; 46(4): 443-9, 1997 Apr.
Article in English | MEDLINE | ID: mdl-9094090

ABSTRACT

Under the influence of progesterone, the porcine uterus synthesizes plasmin/trypsin inhibitor (PTI), a low molecular weight protein (M(r) approximately 14,000) belonging to the Kunitz family of proteinase inhibitors. Here it is demonstrated that mRNA for the same protein is produced by the developing trophoblast during early pregnancy and by the placenta throughout gestation. The transcript for PTI was represented in approximately 0.1% of total phage plaques of a day 13-17 porcine conceptus cDNA library. It shared 99% nucleotide sequence identity with the cDNA isolated from the uterine library and encoded a 112-amino-acid protein identical to the uterine-produced PTI, which has a well-defined Kunitz domain comprised of 64 residues at its amino terminus. Northern analysis and in situ hybridization studies confirmed that expression of PTI by the conceptus begins as early as day 10 of pregnancy and is continued in placental tissues until at least day 90 of gestation. Expression was trophoblast specific at day 20 of gestation, as in situ hybridization detected no mRNA in the embryo. The pattern of PTI expression during pregnancy is consistent with a role either in controlling trophoblast invasiveness or in inhibiting proteinases with trypsin-like specificity released by immune or inflammatory cells.


Subject(s)
Antifibrinolytic Agents/metabolism , Peptides , Plant Proteins , Swine/anatomy & histology , Trophoblasts/enzymology , Trypsin Inhibitors/biosynthesis , Amino Acid Sequence , Animals , Female , In Situ Hybridization , Molecular Sequence Data , Pregnancy , Swine/embryology
9.
J Biol Chem ; 269(39): 24090-4, 1994 Sep 30.
Article in English | MEDLINE | ID: mdl-7929061

ABSTRACT

The porcine uterus synthesizes a proteinase inhibitor (M(r) 14,000) under the influence of progesterone that is relatively specific for plasmin and trypsin, but that also has weak affinity for chymotrypsin. Several isoforms of this uterine plasmin/trypsin inhibitor were purified by a procedure whose final two steps involved affinity chromatography on immobilized chymotrypsin and cation exchange chromatography. Amino-terminal sequencing showed that at least three of the isoforms were closely related. An oligonucleotide probe based on the protein sequence was used to identify a cDNA that contained an open reading frame coding for a mature protein (M(r) 10,295) of 93 amino acids. The inhibitor had a well defined, but unique, Kunitz domain of 64 residues at its amino terminus that shared 67% sequence identity to bovine pancreatic trypsin inhibitor. Its P1 residue was arginine rather than lysine. Northern analysis showed the presence of a single mRNA species (700 bases) that in adult female pigs appeared to be confined to the uterus. During pregnancy, UPTI mRNA expression was high until Day 30 and decreased significantly thereafter. By contrast, uteroferrin mRNA reached maximal concentrations in late pregnancy. These data are consistent with an earlier hypothesis that the inhibitor serves to neutralize the activities of one or more serine proteinases generated by the proliferating trophoblast during the formation of the noninvasive placenta of the pig.


Subject(s)
Peptides , Plant Proteins , Trypsin Inhibitors/metabolism , Uterus/enzymology , Amino Acid Sequence , Animals , Base Sequence , Chromatography, Ion Exchange , Cloning, Molecular , DNA, Complementary , Female , Male , Molecular Sequence Data , Pregnancy , RNA, Messenger/genetics , RNA, Messenger/metabolism , Sequence Homology, Amino Acid , Swine , Trypsin Inhibitors/genetics , Uterus/metabolism
10.
J Anim Sci ; 71(9): 2499-505, 1993 Sep.
Article in English | MEDLINE | ID: mdl-8407662

ABSTRACT

A study was conducted to validate the previously reported growth response to cholecystokinin octapeptide (CCK-8) immunization in barrows and was extended to include gilts. Group-penned barrows and gilts were used to represent conditions in the swine industry. Thirty-two animals, 19 barrows and 13 gilts, were randomly assigned by sex to four pens and two treatments. The control groups were immunized with human serum globulin (hSG). The treated groups (CCK) were immunized with the C-terminal octapeptide of cholecystokinin conjugated to human serum globulin. Specific binding of CCK-8 was confirmed at 29 d after the primary inoculation. Antisera titers were highly variable throughout. The mean titer reached a peak on d 57 and then declined. Body weight gains during the last 49 d, the period during which titers were expressed, were compared by ANOVA. The treatment effect on gain was significant (P = .018); the sex effect approached significance (P = .071); the treatment x sex interaction effect was not significant (P = .82). Least squares mean gain of the CCK group was 8.4% greater than of the hSG group, 41.4 vs 38.2 kg, respectively. A significant linear regression coefficient for gain vs antisera titer was obtained for barrows (P = .03; r2 = .44) but not for gilts. Several carcass variables showed trends similar to that of BW gain, but the treatment effects were less robust (P < .05 to .10). These results generally confirm the findings of the previous study; CCK-8 immunization stimulated growth of barrows by 7.5% in the present and by 10.8% in the previous study.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Immunization/veterinary , Sincalide/immunology , Swine/growth & development , Analysis of Variance , Animals , Antibody Formation , Body Weight , Female , Immune Sera/immunology , Least-Squares Analysis , Male , Meat/standards , Random Allocation , Sex Factors
11.
Biol Reprod ; 48(5): 998-1005, 1993 May.
Article in English | MEDLINE | ID: mdl-8481487

ABSTRACT

Retinol-binding proteins (RBP) are secreted by the porcine uterus under the influence of progesterone and consist of multiple charge forms. Evidence has been previously presented by this laboratory that these uterine RBP are distinct from serum RBP. We have followed the secretion of the uterine RBP during two stages of pseudopregnancy, examined their properties and amino acid sequences, and attempted to clone their cDNA. Analysis of the charge forms present in uterine flushes by anion-exchange chromatography showed that forms 1 (p < 0.01) and 3 (p < 0.05) predominated at Day 13, whereas forms 2 (p < 0.05) and 4 (p < 0.01) were most abundant at Day 45. All four charge forms appeared to form stable complexes with transthyretin (TTR) and were recognized by antiserum to human serum RBP on Western blots. Several cDNA clones isolated from an endometrial cDNA library all appeared to code for a protein identical to classical RBP. Off-blot amino acid sequencing of the first ten residues of two of the more divergent charge forms of uterine RBP indicated complete sequence identity with pig serum RBP. These data suggest that the uterine RBP charge forms may be slightly modified forms of a single protein product corresponding to the classical form of RBP. The change in appearance of the charge forms during pseudopregnancy is probably due to chemical modifications. These modifications do not appear to influence the binding of each charge form to TTR.


Subject(s)
Retinol-Binding Proteins/genetics , Swine/genetics , Uterus/chemistry , Amino Acid Sequence , Animals , Base Sequence , Blotting, Western , Chromatography, High Pressure Liquid , Cloning, Molecular , DNA/chemistry , DNA/genetics , Female , Molecular Sequence Data , Prealbumin/metabolism , Pseudopregnancy , Retinol-Binding Proteins/chemistry , Retinol-Binding Proteins/metabolism , Swine/physiology
12.
J Reprod Fertil Suppl ; 48: 171-86, 1993.
Article in English | MEDLINE | ID: mdl-8145203

ABSTRACT

The second week of pregnancy is a particularly critical period for embryonic survival in pigs. Within that time, conceptus oestrogen synthesis is initiated, spacing and final placement of conceptuses is completed, and the signal for extending the functional lifespan of the corpora lutea is received by the mother. There is also a marked increase in blood flow to the uterus and the uterine endometrium produces and secretes nutrient histotrophe. Conceptus-derived oestrogen has been implicated in many of these events. It is also during this period that the trophoblast elongates dramatically and the inner cell mass starts to differentiate into the embryo proper. Here, we critically review the evidence that oestrogen is the sole factor initiating long-term corpus luteum maintenance in pigs. We also review the functions and general properties of the major secretory proteins in histotrophe and the role of oestrogen in controlling their expression. It is now generally accepted that asynchrony within a litter underlies much of the losses of conceptuses that are otherwise genetically normal, but which are lagging in their development; however, the underlying mechanisms remain unclear. Here we hypothesize that oestrogenic compounds derived from more advanced conceptuses or provided prematurely, either by injection or in the diet, trigger a massive increase in uterine expression and secretion of retinol-binding protein laden with retinol. We propose that less developed, smaller conceptuses are least able to contend with the sudden exposure to this potential teratogen at a time when they are particularly susceptible to imbalance in retinol supply. Hence, even though their growth proceeds for a few days, their developmental potential is irrevocably compromised.


Subject(s)
Corpus Luteum Maintenance/physiology , Estrogens/physiology , Pregnancy, Animal/physiology , Swine/physiology , Animals , Female , Pregnancy , Proteins/metabolism , Trophoblasts/physiology , Uterus/physiology , Vitamin A/physiology
13.
Endocrinology ; 130(5): 2557-64, 1992 May.
Article in English | MEDLINE | ID: mdl-1572282

ABSTRACT

The endometrium of the pig secretes retinol-binding protein (RBP) under the influence of progesterone (P4). The objective of this study was to determine how conceptus-derived estrogen might modulate this production of RBP around days 11-13 of pregnancy when conceptuses elongate from spheres to long thread-like forms. Concentrations of retinol and RBP were low (35 +/- 7 ng/ml) in uterine flushings obtained on days 10-12 of the estrous cycle or from pregnant gilts in which conceptuses had not elongated. Concentrations of retinol and RBP increased 7- to 8-fold (P less than 0.01) in flushings where filamentous conceptuses were present. Size exclusion and ion exchange chromatography demonstrated that virtually all retinol assayed in uterine flushings was associated with RBP. Northern blot analysis with a cDNA representing uterine RBP revealed a single endometrial mRNA 1.1 kilobases in length. Expression of RBP mRNA in uterine endometrium was measured in ovariectomized prepubertal gilts after the administration of steroids according to the following regimens: I, corn oil (days 0-16; n = 10); II, estradiol benzoate (EB; days 13-14; n = 11); III, EB (days 1-2; n = 12); IV, EB (days 1-2) plus P4 (days 3-16; n = 12); and V, EB (days 1-2) plus P4 (days 3-16) plus EB (days 13-14; n = 12). EB (200 micrograms) and P4 (100 mg) were administered twice daily. Treatment IV was designed to stimulate the estrous cycle, and treatment V simulated early pregnancy. All gilts were hysterectomized on day 16, and total uterine mRNA (3 micrograms) was analyzed by Northern blotting. No RBP mRNA was detected in groups I, II, or III. In group IV, 5 of 12 gilts had detectable RBP mRNA, as measured by densitometric scanning (OD = 0.35 +/- 0.14). RNA isolated from all gilts in group V (12 of 12) gave a strong hybridization signal (OD = 1.58 +/- 0.22) for RBP. Finally, RBP mRNA was examined in the uterine endometrium of mature gilts on day 13 of the estrous cycle (n = 4), day 13 of pseudopregnancy (2.5 mg EB given on days 11-12; n = 4), or day 13 of pregnancy after conceptuses had elongated (n = 4). RBP mRNA was present in all groups, but was enhanced approximately 12-fold (P less than 0.01) in pregnant and pseudopregnant gilts compared to that in control gilts.(ABSTRACT TRUNCATED AT 400 WORDS)


Subject(s)
Endometrium/metabolism , Estradiol/pharmacology , Pregnancy, Animal/metabolism , Progesterone/pharmacology , Retinol-Binding Proteins/biosynthesis , Uterus/metabolism , Vitamin A/metabolism , Animals , Chromatography, High Pressure Liquid , Endometrium/drug effects , Estrus/drug effects , Estrus/metabolism , Female , Ovariectomy , Pregnancy , RNA, Messenger/metabolism , Reference Values , Retinol-Binding Proteins/genetics , Retinol-Binding Proteins/metabolism , Swine , Uterus/drug effects
14.
J Reprod Fertil ; 94(1): 107-14, 1992 Jan.
Article in English | MEDLINE | ID: mdl-1552470

ABSTRACT

Changes in plasma and follicular fluid concentrations of inhibin were examined in sows after weaning at 28-32 days post partum. From 0 to 48 h after weaning, inhibin concentrations were 200-300 times higher in follicular fluid from small (less than 4 mm) and medium-large (greater than or equal to 4 mm) follicles than in ovarian venous plasma. Inhibin concentrations increased in follicular fluid from medium-large follicles at 24 and 48 h after weaning; concentrations in ovarian venous plasma were positively correlated with the number of medium-large follicles (r = 0.40) and with ovarian venous plasma concentrations of oestradiol (r = 0.61). Blood samples were collected for 30 days from sows (n = 6) that exhibited oestrus within 5 days after weaning and from sows (n = 5) that remained anoestrous for 11 days after weaning. Plasma inhibin concentrations rose in oestrous and anoestrous sows by 12 h and continued to rise for 60 h after weaning. Plasma inhibin concentrations rose further and were higher at 3.5-4.5 days after weaning in oestrous sows than in sows that remained anoestrous. After oestrus, plasma inhibin concentrations declined. At weaning, plasma concentrations of follicle-stimulating hormone (FSH) were higher in sows that subsequently exhibited oestrus than in sows that remained anoestrous. After weaning, plasma concentrations of FSH declined in both groups, reached a nadir at 2.5 days, and increased gradually in anoestrous sows; oestrous sows exhibited an FSH surge at oestrus. Plasma FSH returned to preweaning concentrations in both groups of sows at Days 7-8.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Estrus/blood , Follicular Fluid/chemistry , Inhibins/analysis , Swine/metabolism , Weaning , Animals , Estradiol/blood , Female , Follicle Stimulating Hormone/blood , Follicular Phase/physiology , Inhibins/blood , Ovary/blood supply , Progesterone/blood
16.
Mol Endocrinol ; 5(10): 1533-40, 1991 Oct.
Article in English | MEDLINE | ID: mdl-1723146

ABSTRACT

Retinol-binding protein (RBP) is a major secretory product of the porcine conceptus. Using an oligonucleotide probe corresponding to a highly conserved region of all known mammalian RBP, we have isolated an apparently full-length cDNA clone for porcine conceptus RBP from a cDNA library constructed from pig conceptuses collected between days 13-17 of pregnancy. The cDNA was 937 base-pairs in length and coded for a protein whose inferred amino-terminal sequence was identical to that reported for both porcine conceptus RBP and porcine serum RBP. Its length was consistent with the size (approximately 1 kilobase) of the RBP message in porcine conceptuses. Porcine conceptus RBP and human serum RBP share 91% amino acid sequence identity. The inferred differences in sequence were evenly distributed throughout the length of the polypeptide. RBP mRNA was detectable within the trophoblast of day 11 porcine conceptuses by in situ hybridization with a 618-basepair 35S-labeled probe corresponding to the 3' end of porcine RBP. Silver grain density was distributed relatively uniformly over the trophoblast and the inner cell mass. Western blot analysis of conceptus culture medium demonstrated that the conceptuses of cattle (on day 19) and sheep (on day 15) as well as pigs secrete RBP during early pregnancy. Secretion of large quantities of RBP by the trophoblast of preimplantation pig conceptuses suggests important roles for vitamin A and RBP near the time of conceptus elongation.


Subject(s)
Ectoderm/physiology , Embryo, Mammalian/physiology , Liver/physiology , Retinol-Binding Proteins/genetics , Swine/physiology , Amino Acid Sequence , Animals , Base Sequence , Blotting, Northern , Cattle , Cloning, Molecular/methods , DNA/genetics , DNA/isolation & purification , DNA Probes , Ectoderm/cytology , Embryo, Mammalian/cytology , Gene Expression , Gene Library , Humans , Molecular Sequence Data , Nucleic Acid Hybridization , Organ Culture Techniques , RNA/genetics , RNA/isolation & purification , RNA, Messenger/analysis , RNA, Messenger/genetics , Sequence Homology, Nucleic Acid , Sheep , Swine/embryology
17.
J Endocrinol ; 125(1): 123-9, 1990 Apr.
Article in English | MEDLINE | ID: mdl-1971003

ABSTRACT

Steers were actively immunized at 81 days of age against human serum albumin (hSA; controls) or hSA conjugated to either somatostatin (SRIF) or growth hormone-releasing factor (GRF). Binding titres were observed for the respective peptide antigens after all steers had been given booster immunizations. Although no effects of treatment were observed in SRIF-immunized steers, mean serum concentrations of GH and insulin-like growth factor (IGF-I) were suppressed (P less than 0.01) in GRF-immunized steers when compared with hSA-immunized controls. Mean concentrations of prolactin did not differ with treatment but showed seasonal fluctuations (P less than 0.001) associated with changes in the daylength. In contrast to its marked effect upon serum concentrations of IGF-I, immunization against GRF resulted in a relatively small (6%) but significant decrease in body weight gain (P less than 0.01) and an increase in carcass backfat thickness (P less than 0.05). In summary, our findings have shown the susceptibility of steers to growth modulation by GRF immunoneutralization. Secondly, the poor relationship observed between serum concentrations of IGF-I and growth rates in GRF-immunized steers suggested that circulating IGF-I may not be the principle factor determining the post-weaning growth rate in cattle.


Subject(s)
Growth Hormone-Releasing Hormone/immunology , Growth Hormone/blood , Growth/immunology , Insulin-Like Growth Factor I/metabolism , Somatomedins/metabolism , Somatostatin/immunology , Animals , Cattle , Immunization , Male
18.
J Biol Chem ; 265(6): 3248-55, 1990 Feb 25.
Article in English | MEDLINE | ID: mdl-2303448

ABSTRACT

Two-dimensional polyacrylamide gel electrophoresis has revealed the presence of a group of relatively acidic proteins of molecular weight about 22,000 in the uterine flushings of pseudopregnant pigs. The proteins have been purified by a combination of gel filtration chromatography and high performance anion-exchange chromatography and shown to bind both [3H] retinol and [3H]retinoic acid. At least four protein peaks that bound retinoids could be detected in the uterine secretions of a single pig. The ion-exchange procedure also allowed the retinol-free apoproteins to be separated from the holoforms that had associated ligand. Amino acid sequencing of the NH2 termini of polypeptides within three of the peaks revealed the presence of proteins with some degree of sequence identity to serum retinol-binding proteins (RBP). The most basic polypeptides showed the least similarity (about 30% identity), while the most acidic isoform analyzed shared about 70% sequence identity with the NH2 terminus of human serum RBP. Western blotting procedures employing an antiserum raised against the most basic isoforms showed that the amount of retinol-binding protein in uterine secretions increased markedly in ovariectomized animals in response to long term progesterone treatment. These proteins appear to form part of the uterine histotroph thought to be essential for nourishment of the conceptuses during pregnancy. A simple three-step procedure for purifying retinol-binding protein from pig serum is also described. The NH2-terminal sequence of this RBP is similar to that of human RBP but different from those of the uterine forms. The study suggests that a family of RBP, distinct from the serum form, is secreted by the uterine endometrium of the pig in response to progesterone.


Subject(s)
Endometrium/metabolism , Progesterone/pharmacology , Retinol-Binding Proteins/biosynthesis , Uterus/metabolism , Amino Acid Sequence , Animals , Cells, Cultured , Chromatography, Gel , Chromatography, High Pressure Liquid , Chromatography, Ion Exchange , Electrophoresis, Gel, Two-Dimensional , Endometrium/drug effects , Female , Humans , Molecular Sequence Data , Molecular Weight , Ovariectomy , Progesterone/blood , Retinol-Binding Proteins/genetics , Retinol-Binding Proteins/isolation & purification , Sequence Homology, Nucleic Acid , Swine , Uterus/drug effects , Vitamin A/metabolism
19.
Growth Dev Aging ; 54(1-2): 51-6, 1990.
Article in English | MEDLINE | ID: mdl-2210922

ABSTRACT

Experimental superalimentation at 30% above ad libitum intake increased growth 40% and confirmed that voluntary food intake is a growth-limiting factor in swine. A sequence of contingent hypotheses was proposed for swine: cholecystokinin (CCK) is a regulator of food intake; food intake is enhanced by reduction of serum CCK; serum CCK is reduced by anti-CCK antibodies: anti-CCK antibodies are raised by active immunization. The objectives of this study were to determine if antibodies were raised in immunized swine and if the anti-CCK titers were sufficient to increase food intake and growth. Twelve young growing swine were immunized against cholecystokinin (CCK-8) to test the hypothesis that anticholecystokinin antibodies in serum would suppress cholecystokinin inhibition of appetite (food intake). An equal number of control animals (hSG) were immunized against the antigenic carrier protein by the same protocol. Specific binding of [125I]CCK-8, the C-terminal octapeptide, by sera diluted 1:181 increased to a peak value on day 43. Food intake and body weight gain were similar for the two groups during the first phase of the study. However, food intake was 8.2% greater and body weight gain was 10.6% greater for the CCK-8 than for the hSG group during the second phase (d 43 to d 77). Total food intake over the 77-day study was 5.4% greater for the CCK-8 group (P = .08): body weight gain was 8.3% greater (P = .006). Regression analyses confirmed that gain over the 34-day second phase increased .076 kg (P = .045; R2 = 0.34) for each percentage unit increase of serum binding.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Cholecystokinin/physiology , Eating/physiology , Swine/physiology , Animals , Appetite/physiology , Cholecystokinin/antagonists & inhibitors , Cholecystokinin/immunology , Immunization , Male , Swine/growth & development , Weight Gain/physiology
20.
J Anim Sci ; 67(10): 2709-14, 1989 Oct.
Article in English | MEDLINE | ID: mdl-2808172

ABSTRACT

This study explored feed intake and carcass responses to active immunization against desulfated cholecystokinin-octapeptide (CCK-8) in ram lambs. Antibody titers 8 wk following primary immunization and booster immunizations given at 4 and 6 wk averaged greater than 1:1,000. Titers increased to greater than 1:10,000 by 16 wk following a final booster immunization at 11 wk. The antibodies developed against desulfated CCK-8 exhibited 29% and 13% cross-reactivities for sulfated CCK-8 and gastrin-17, respectively. Immunization against desulfated CCK-8 had no effect on feed intake, ADG, carcass weight or carcass quality grade. Backfat thickness and carcass yield grade were reduced (P less than .05) by immunization. Organ weights at slaughter, including those of the pancreas and small intestines, were not affected by CCK-8 immunization, with the exception of the lungs, which were 16% lighter (P less than .01) in immunized lambs. In conclusion, active immunization against desulfated CCK-8 resulted in development of high antibody titers against desulfated and sulfated CCK-8. Immunization against CCK-8 decreased fat content of the carcass but failed to affect feed intake, carcass weight or ADG.


Subject(s)
Eating , Immunization/veterinary , Sheep/growth & development , Sincalide/immunology , Weight Gain , Animals , Antibody Formation , Body Composition , Cross Reactions , Immunization, Secondary/veterinary , Male , Organ Size , Sheep/immunology
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