Fate of hamster oviductin in the oviduct and uterus during early gestation

Oviductins are a family of glycoproteins which are synthesized and secreted by oviductal secretory cells and which, upon their secretion in the lumen of the oviduct, become associated with postovulatory oocytes and developing embryos. Recently, we showed that hamster oviductin is maximally secreted in the oviduct at the time of ovulation and is later associated with a certain population of uterine epithelial cells, where it is subsequently endocytosed and degraded. In light of these results, this study was conducted to follow the fate of hamster oviductin in the oviduct and uterus during early gestation. Using a monoclonal antibody against hamster oviductin, immunofluorescence and immunogold labeling revealed that during early gestation, immunoreactivity to oviductin in the uterus gradually diminished to an almost total disappearance at time of implantation. However, the strong labeling intensity remained unchanged in the oviduct. Biochemical analyses demonstrated that a degradation of oviductin occurs in the uterus, and a loss of immunoreactivity was also observed as gestation progressed, so that by the time of implantation, immunoreactivity to oviductin was barely detectable. The decrease of oviductin along the uterine epithelium at the time of blastocyst attachment and its final disappearance at implantation suggest that this glycoprotein could be a potential modulator of uterine receptivity.     

Fluoride in bone of rats anesthetized during gestation with enflurane or methoxyflurane

1. The interaction between the reversible cholinesterase inhibitors, physostigmine and neostigmine and reserpine was studied. 2. A combination of reserpine plus a cholinesterase inhibitor significantly increased lethality in adult male Swiss-Webster mice above that caused by either neostigmine, physostigmine, or reserpine alone. 3. Methscopolamine completely reversed this effect. 4. It would appear that the presence of the antiadrenergic agent, reserpine, increases the toxicity of cholinomimetic agents. 5. The above results may have clinical significance, since reserpine and cholinesterase inhibitors are used in the practice of medicine.     

Effects of anandamide on gestation in pregnant rats

The present study was to test whether the recently described endogenous ligand for the cannabinoid receptor; arachidonyl-ethanolamide (anandamide, ANA), may produce similar effects on pregnancy as the main psychoactive component of marihuana: delta9-tetrahydrocannabinol (THC) in rats. ANA, THC (0.02 mg/kg i.p./day, respectively) or vehicle were injected daily over the third week of pregnancy. The pregnant rats were either killed on day 21 of pregnancy or followed up to delivery. Results show a significant increase in the duration of pregnancy after both THC and ANA treatment. Both drugs caused an increase in the frequency of stillbirths. The mothers' hormone contents in tissues and sera were measured. Decreased LH content was observed in the serum of treated animals. No changes in FSH content were observed either in the pituitary or in the sera. Pituitary prolactin (PRL) levels was lower in ANA treated animals as compared both to controls or THC treated subjects. The serum PRL content decreased in all experimental groups. Decrease in serum progesterone was more prominent in treated rats. Serum levels of prostaglandins (PGF 1alpha and PGF 2alpha) were significantly decreased after THC and ANA treatment. We conclude that ANA has the same tendency to change reproductory parameters in pregnant rats as THC, although in some cases the effects of ANA were slightly different from that of THC. Both endogenous and exogenous cannabinoids inhibit PG synthesis in pregnant rats and this maybe responsible for the delay constitute the mechanism in the onset of labour.     

Effect of cholinesterase inhibitor malathion on whole body irradiated rats

Under physiological conditions, erythrocytes of the horse metabolized 638 +/- 37 (+/-SE) nmoles glucose/ml cells/hr at 37 degrees C compared to 942 +/- 31 for the cat, 1,329 +/- 44 for the dog, and 1,485 +/- 43 for man. On an absolute basis, pentose phosphate metabolism was similar between species, with species differences in erythrocyte glucose tulization attributable to differences in Embden-Meyerhof pathway metabolism. By examining pentose phosphate pathway recycling, it appears that some functional compartmentation exists within erythrocytes.     

Dietary thiamin supply during gestation effects thiamin status of lactating rats and their suckling offspring

A paired-comparison study of manual and automated (PAPNET) screenings of cervico-vaginal smears (Pap tests) was conducted to determine whether primary PAPNET screening was a reliable alternative to manual screening. A series of 5,037 consecutive Pap tests was first screened by the manual method. Next they were scanned by the PAPNET system, the DAT tapes were reviewed, and using a nonspecific triage protocol, abnormal tests were identified for limited, manual rescreening. False-negative rates (FNR) for each method were calculated and analyzed for due cause. By manual and PAPNET screening, respectively, there were 436 (8.6%) and 250 (4.9%) abnormal results. Manual screening missed 18 abnormals (5 SIL) and PAPNET 202 (7 SIL). The primary, manual screenings relating to the PAPNET false-negative tests were reviewed and revised to normal in 30. Based on the changes in the other 172 tests, cellular features ostensibly missed by the PAPNET system were listed to form part of a specific triage protocol, and were used to scrutinize the companion 172 DAT tapes: 150 tapes were abnormal. The manual FNR for an abnormal (SIL) result was 4% and (8.8%), respectively. Equivalent FNR pre- and postreviews for the PAPNET system were 44.6% (10.6%) and 5.2% (1.3%), respectively. This study discovered that the evaluation of some cell features in monitor-based, video images was the most important factor limiting the application of the PAPNET system as a primary screener. When governed by a specific triage protocol incorporating these features, primary PAPNET screening has the potential to equal the laboratory threshold of manual screening and be a better detector of SIL.     

Bioavailability in rats of bound pesticide residues from tolerant or susceptible varieties of soybean and canola treated with metribuzin or atrazine

Percutaneous treatments are useful in two, frequent, painful conditions involving the shoulder. In frozen shoulder syndrome, distention arthrography with intra-articular injection of steroid is used to provide pain relief and to improve joint motion. In rotator cuff tendon calcifications, needle aspiration of calcific deposits is used to treat pain. Surgery should be restricted to failures of needle aspiration. The techniques of these procedures are described and their results are reported.     

Fate of antibodies bound to lymphocyte surface. II.--Degradation and release of immune complexes

The fate of horse anti-human lymphocyte globulins, rabbit anti-human beta2 microglobulin antibodies and anti-HL-A alloantibodies radiolabelled with 125I and complexed to their corresponding lymphocyte surface antigens, was investigated. During prolonged incubation at 37 degrees C, part of these antibodies were actively released into the surrounding medium after enzymatic degradation occuring either intracellularly or at the cell surface. Metabolism of the antibodies was temperature-dependent and involved only antibodies able to react with antigens on the cell surface. Sephadex G200 filtration of the small percentage of non dialysable radioactive products in the supernatant revealed that all the 125I-anti-human lymphocyte globulin was recovered as a macromolecular structure in the void volume. Similar experiments were performed with 51Cr labelled lymphocytes coated with anti-lymphocyte globulins: treatment with anti-horse immunoglobulin precipitated part of 51Cr activity in the void volume which was therefore associated with anti-human lymphocyte globulin as an immune complex. The same protocol applied to the other two antibodies studied showed that anti-beta2 microglobulin antibodies were recovered both as macromolecular and 7 S immune complexes, whereas anti-HL-A antibodies were recovered as macromolecular, 7 S and still smaller immune complexes.     

Prenatal malnutrition-induced functional alterations in callosal connections and in interhemispheric asymmetry in rats are prevented by reduction of noradrenaline synthesis during gestation

Prenatal malnutrition results in increased concentration and release of central noradrenaline, a neurotransmitter that is an important regulator of normal regressive events such as axonal pruning and synaptic elimination. This suggests that some of the functional disturbances in brain induced by prenatal malnutrition could be due at least in part to increased noradrenaline activity that may enhance regressive events during early stages of development. To test this hypothesis we studied whether chronic administration of alpha-methyl-p-tyrosine, an inhibitor of tyrosine hydroxylase, to rats during gestation might prevent long-term deleterious effects of prenatal malnutrition on functional properties of interhemispheric connections of the visual cortex, and on asymmetry of visual evoked responses. The experiments were conducted on normal and malnourished rats 45-50 d of age. Prenatal malnutrition was induced by restricting the food consumption of pregnant rats to 40%, from d 8 postconception to parturition. At birth, prenatally malnourished rats had significantly greater whole-brain noradrenaline concentration as well as significantly enhanced noradrenaline release in the visual cortex. At 45-50 d of age, the malnourished group had a significantly smaller cortical area, exhibiting transcallosal evoked responses; in addition, the amplitude of these responses was significantly smaller. Malnourished rats showed a significant reduction of the normal interhemispheric asymmetry of visual evoked responses. The addition of 0.3% alpha-methyl-p-tyrosine to the diet of malnourished pregnant rats during the last 2 wk of gestation prevented functional disorders induced in the offspring by prenatal malnutrition on interhemispheric connectivity of visual areas and on interhemispheric bioelectrical asymmetry, probably by reducing the elevated brain noradrenaline activity and thereby restoring the normal trophic role of this neurotransmitter.     

Site specificity of surfactant protein expression in airways of baboons during gestation

BACKGROUND: There are disparate reports concerning the presence of surfactant proteins in the airways of lung. The recent finding of SP-A in tracheobronchial epithelium and submucosal glands in lungs from second trimester humans has renewed interest in potential new functions of surfactant in lung biology. METHODS: In situ hybridization studies were done to determine the distribution of SP-A, SP-B, and SP-C in baboon lung specimens from 60, 90, 120, 140, 160, and 180 (term) days of gestation and adults. Lungs from gestation controls were obtained at the time of hysterotomy and adult lungs at necropsy. Riboprobes used for in situ hybridization contained the entire coding regions for human SP-A, SP-B, and SP-C. RESULTS: At 60 days, SP-C mRNA expression was evident in focal portions of primitive tubular epithelium but not bronchi. This distal pattern of SP-C mRNA expression persisted and was present in some epithelial cells of respiratory bronchioles at term. At 90 days, SP-A mRNA expression was present in the epithelium of trachea and large bronchi. SP-B mRNA expression was found in small bronchi, bronchioles, and distal tubular epithelium at 120 days of gestation. SP-A mRNA bronchiolar localization became evident at 140 days of gestation and alveolar type 2 cellular expression at 160 days of gestation. Abrupt transitions of surfactant protein expression were identified (e.g., SP-A mRNA-positive cells in the epithelium of large bronchi with adjoining SP-B mRNA expression in small bronchi and bronchioles). CONCLUSIONS: Findings in the baboon indicate that there are well-delineated sites of surfactant protein mRNA expression in bronchial and bronchiolar epithelia. mRNA expressions of SP-A and SP-B are present in both bronchial and bronchiolar epithelium but at different sites, whereas SP-C expression is seen in loci of epithelial cells in respiratory bronchioles.     

Degradation of malathion in thermally generated aerosols

The degradation of malathion, undiluted and diluted with No. 2 fuel oil, in thermally generated aerosol clouds was examined at selected temperatures ranging from 121 to 566 degrees C. Undiluted malathion residues decreased from 1.76 to 0.21 micrograms/ml over this range of temperatures. Malathion diluted with fuel oil decreased from 0.14 to 0.02 microgram/ml as the temperature was increased 288 to 510 degrees C.     

Serial measurement of arterial plasma progesterone levels throughout gestation and parturition in individual rats

Daily blood samples were taken from 6, chronically cannulated, fully conscious rats to measure plasma progesterone levels throughout gestation. Progesterone levels in individual rats fluctuated by up to 28 ng/ml per day, but tended to be consistently higher or lower than the group mean. The accuracy of predicting progesterone levels in individual rats from previous values was examined. Progesterone levels on day 7 of gestation were negatively correlated with foetal weights near term. There was little indication that high progesterone levels at any stage of gestation lead to increased foetal or placental weights. Progesterone levels on day 17 were positively correlated with the number of corpura lutea but there was little relationship between progesterone and either the number or total mass of the placentas. Serial blood samples taken from a second group of 6 rats at 2 hourly intervals showed that the time between the major fall in progesterone levels to below 12 ng/ml and the onset of parturition was relatively constant (varying by only 8 h) despite a 29 h range in the total length of gestation.     

Ultrastructural observations of the human uterine smooth muscle cells during gestation

At term pregnancy, the myometrium consists of bundles of smooth muscle cells bound together by varying amounts of connective tissue. Each bundle contains both dark and light muscle cells. During uterine contractions it is believed that the smooth muscle cells become darker, decrease in volume, and exhibit changes in diameter. This is accompanied by widening of the interspaces and by a decrease in the areas of cellular contact. Between contractions, there are more light cells which become arranged closer to each other and exhibit large areas of interdigitation. The significance of these observations in the mechanism of uterine contraction and retraction is discussed. Cell believed to be modified smooth muscle cells occupy the myoendometrial junction and the decidua basalis. They are irregular in shape, poor in myofilament content, and rich in other cytoplasmic organelles and form a loosely arranged layer of cells between the myometrium and the trophoblast. The possible functional significance of these cells is also discussed.     

Doppler ultrasonography of canine maternal and fetal arteries during normal gestation

Two-dimensional ultrasonography was used in combination with colour-flow imaging and pulsed wave Doppler ultrasonography to study the maternal circulation and the development of fetal vascularization in six Beagles during normal gestation. For the first time, the development of the circulation was demonstrated in the bitch and her fetuses intra vitam. The bloodstream was examined in small uteroplacental arteries, the umbilical artery, the fetal aorta and the common carotid artery. The duration of the study was from week 3 after insemination until birth. Relatively large vessels were detected by cross-sectional ultrasonography, and small vessels were detected by colour-flow imaging. In pulsed wave Doppler ultrasonography, the blood flow was measured and described using the parameters of systolic peak velocity, diastolic peak velocity, end-diastolic velocity, pulsatility index, resistance index, A:B ratio (systolic peak velocity:end-diastolic velocity) and S:D parameter (systolic peak velocity:diastolic peak velocity). The development of the measured parameters is typical and similar to that in humans. The systolic peak velocity of the canine maternal uteroplacental arteries shows important differences in comparison with humans. The pulsatility index, resistance index and A:B ratio decrease in nearly all vessels. Only the fetal common carotid artery has constant pulsatility and resistance indices during gestation. For the first time, the quality and quantity of the normal blood flow have been monitored during the whole of gestation. A normal circulation is fundamental for supplying the fetus adequately with oxygen and nutrients and thus for physiological development. These ultrasonographic results are the basis for further clinical studies.     

Morphological study of endothelial cells in the human fetus during early period of gestation

During the formation of blood vessels, proliferation and migration of endothelial cells (ECs) are one of the most important mechanisms. The development of the vessels involves two different mechanisms: vasculogenesis i.e. differentiation of ECs in situ from mesenchymal precursors, and angiogenesis i.e. sprouting of ECs from pre-existing vessels. The frontal lobes from 20 brains of human fetuses ranging from 8 to 17 weeks of gestation (GW) were subjected to correlative light and electron microscopic studies. Our observations demonstrate the sprouting of ECs from pre-existing vessels in leptomeninges (angiogenesis) during the formation of a capillary network of the fetal human brain. In addition, the study did not reveal after the 8th GW the mitotic proliferation of ECs in examined specimens which allows to underline the importance of sprouting and elongation of ECs channels for this phase of vascularization of cerebral hemispheres.     

Trophoblast cell-mediated modifications to uterine spiral arteries during early gestation in the macaque

A specialized subset of invasive embryonic cytotrophoblast cells gains access to maternal uterine arteries early in the gestation of higher primates. These cells continue to migrate extensively within the lumina of spiral arteries, converting them to the highly modified uteroplacental arteries of pregnancy. Although trophoblast cell-mediated modifications are considered critical to the progress of normal pregnancy, few studies have addressed the cellular interactions between maternal arteries and embryonic cells in situ. Macaque placentas and endometrial tissues were collected from 12 animals from day 14 of gestation (blastocyst implantation begins on day 9) to day 49. Standard indirect immunoperoxidase methods were used to identify matrix metalloproteinases (MMP-1, MMP-3, MMP-9), cathepsin B, cathepsin D, platelet-endothelial cell adhesion molecule, cytokeratins, smooth muscle actin, CD68, and factor VIII-related antigen. Cytotrophoblast cells were located deep within spiral arteries in each of the specimens examined. In some examples tightly packed clusters of cytotrophoblast occluded the lumina of invaded arteries. Initial penetration of arterial tunica intima was revealed by discontinuities in the staining pattern for factor VIII and cytotrophoblast intrusion was indicated by cytokeratin staining of the trophoblast cells. Continued cytotrophoblast intrusion into the tunica media was apparent by gaps in the smooth muscle. MMP-1, MMP-3, and MMP-9 were localized within intraluminal and intramural cytotrophoblast. By contrast, neither cathepsin B nor cathepsin D were present, although both were seen in uterine macrophages and stromal cells. Upon reaching the surrounding uterine stroma the cytotrophoblast cells ceased migration. As cytotrophoblast accumulated in the arterial wall the vascular lumen expanded. Evidence of cell death was rarely encountered in associated maternal or embryonic tissues. We conclude that intra-arterial cytotrophoblast cells express several proteinases with substrate specificities sufficient to permit independent remodeling of the extracellular matrix comprising uterine artery walls. The remodeling of the arteries, which involves extensive displacement of maternal endothelium and smooth muscle, in addition to degradation and synthesis of extracellular matrix, is accomplished with little evidence of cell death or loss of the integrity of the arteries. This process provides an interesting example of cooperation between different types of interacting tissues from genetically distinct individuals.     

Heme oxygenase and nitric oxide synthase in the placenta of the guinea-pig during gestation

Two low-molecular cytolytic toxins (RmI and RmII) and four trypsin inhibitors were isolated from the aqueous extract of sea anemone Radianthus macrodactylus. The method of isolation involved precipitation with acetone, gel filtration on acrylex P-4, ion-exchange chromatography on CM-32 cellulose, affinity chromatography on trypsin-binding sepharose 4B, ion exchange chromatography on an Ultrapore TSK CM-3SW column, and reversed phase HPLC on a Silasorb C18 column. RmI, RmII, and JnI inhibitor displayed molecular masses 5100, 6100, and 7100 Da, respectively, when subjected to SDS-PAGE. The isoelectric points were 9.2 and 9.3 for RmI and RmII, respectively. The amino acid composition and N-terminal amino acid residue (glycine) were determined for RmI, RmII, and JnI. Both proteins were nontoxic to mice and crabs. Hemolytic activity was determined to be 25 and 20 HU/mg for RmI and RmII, respectively, and their action on erythrocyte membrane was not inhibited by exogenous sphingomyelin. RmI and RmII exhibited antihistamine activity.     

Frequencies of fetal nucleated red blood cells in maternal blood during different stages of gestation

High molecular weight forms of tyrosinase have been found to be expressed during spontaneous remelanization of the amelanotic B-16 melanoma cells in culture as well as in melanotic tumors formed from amelanotic melanoma cells grown in C57BL/6J mice. Overnight extraction of the crude melanosomal fractions from such tumors and cultured melanoma cells reveal the presence of an additional DOPA-MBTH positive band well below the stacking gel. This band has been found to be alpha-PEP7 (antibody specific for tyrosinase) positive and alpha-PEP1 (antibody specific for TRP-1) negative on Western blot analysis. Heat treatment at 60 degrees C for 60 min results in the loss of this band and considerable loss of activity of the melanosomal extract. Trypsin treatment of these melanosomal extracts resulted in a minor change in the mobility of the high molecular weight band. SDS-PAGE under reduced conditions followed by Western blotting revealed that the high molecular weight band was lost and not detected by alpha-PEP7 or alpha-PEP1. These findings indicate that high molecular weight, heat sensitive and trypsin resistant forms of tyrosinase are transiently expressed in B-16 melanoma cells and tumors that are initiating remelanization following phenotypic drift towards the amelanotic state.     

Changes in kinetic properties of oxytocin receptors in longitudinal muscle membranes of rat uterus during gestation

A minimally invasive method for the stimulation of the spinal cord in dogs was developed. Electrical stimuli were delivered to the spine at the T8 vertebral level through partially insulated needles and under ketamine anesthesia. This allowed the recording of reproducible responses in hind limb muscles of intact dogs. Chronic unilateral deafferentation suppressed the muscle responses on the operated side. A cordotomy sparing the dorsal columns at the L1 level did not completely suppress the muscle responses. It was concluded that motoneuron activation through antidromic conduction in first-order sensory neurons was possible with thoracic spine stimulation and that the recording of muscle responses did not necessarily assess central motor pathways.     

Reactivity of gemfibrozil 1-o-beta-acyl glucuronide. Pharmacokinetics of covalently bound gemfibrozil-protein adducts in rats

Acyl glucuronides are electrophilic metabolites that are readily hydrolyzed, undergo intramolecular rearrangement, and mediate the covalent binding of many acidic drugs to endogenous proteins. Gemfibrozil is extensively metabolized to gemfibrozil acyl glucuronide in humans and rats. The aims of this study were to demonstrate the reactivity of gemfibrozil glucuronide, determine whether gemfibrozil formed covalently bound protein adducts in vivo, describe the pharmacokinetics of adduct formation, and examine the role of gemfibrozil glucuronide in adduct formation. Rats were administered 150 mg/kg gemfibrozil daily for up to 37 days and killed 1, 2, 5, 10, 19, and 37 days after commencement of dosing, and 1, 2, 3, 8, 17, and 30 days after cessation of dosing. Plasma, liver, kidney, and heart were examined for adduct formation. Plasma was quantitatively the most important site for formation of gemfibrozil-protein adducts with mean (SE) steady-state concentrations of 31.40 (2.40) ng/mg protein attained by approximately the 10th day of dosing. Adduct half-life in plasma was 3.1 days, consistent with the elimination half-life of albumin. Mean (SE) kidney, liver, and heart steady-state adduct concentrations were 2.13 (0.11), 0.89 (0.35), and 0.95 (0.07) ng/mg protein, respectively. The rate of gemfibrozil-protein adduct accumulation seemed greatest in liver, but was similar in kidney and plasma, with approximately 2x, 16x, and 30x accumulation, respectively, over the dosing interval. In all tissues, adduct half-lives were significantly greater than those of the noncovalently bound gemfibrozil or gemfibrozil glucuronide.(ABSTRACT TRUNCATED AT 250 WORDS)