Studies on the equine placenta II. Ultrastructure of the placental barrier.

In early pregnancy the equine placenta consists of a simple apposition of fetal and maternal epithelia, but it becomes more complex with the formation of microcotyledons between 75 and 100 days of gestation. Although the placental barrier maintains an epitheliochorial arrangement throughout the course of pregnancy, a thinning of the maternal epithelium and a progressive indentation of the chorionic epithelium by fetal capillaries shortens the length of the diffusion pathway and reduces the amount of placental tissue between fetal and maternal bloodstreams. These structural modifications may reflect the changing requirements of the fetus for O2 and other metabolites as gestation proceeds. During the first 200 days of pregnancy there is evidence of intense pinocytotic activity by the cells of the trophoblast. From the 100th day of pregnancy there is a pronounced development of smooth endoplasmic reticulum, while rough endoplasmic reticulum and irregular, dense, membrane-bound bodies are a prominent feature of the paranuclear cytoplasm from Day 200. These changes suggest that the cells of the trophoblast become more highly involved in synthetic processes with increasing gestational age.     

Studies on the equine placenta. III. Ultrastructure of the uterine glands and the overlying trophoblast

Ultrastructural studies of the uterine glands at intervals during pregnancy in the mare show that secretory activity continues after formation of the placental exchange units. The nature of the glandular secretion appeared initially to be proteinaceous, but cellular debris was also present during the last third of gestation. These secretions were absorbed by the trophoblast overlying the mouths of the glands. The fate secretions and their significance for the fetus and placenta are unknown.     

Ultrastructural studies of the equine uterus and placenta following parturition.

Post-partum placentae and uterine biopsy samples from mares after normal and abnormal foalings are described. After normal delivery there is little damage to fetal or maternal tissues. The villous epitheliochorial palcenta separates cleanly at the maternal-fetal interface and the afterbirth consists almost exclusively of fetal tissue. Uterine involution is well advanced by the 3rd and 4th days post partum and the changes are usually complete by the oestrus 7--10 days after parturition. Placental separation and involution of the uterus appear to proceed normally in malpresented foals and in otherwise viable foals with musculoskeletal defects. In aborted, stillborn or dysmature foals there are obvious signs of damage to both fetal and maternal tissues. It is generally accepted that the growth and development of the fetus is dependent upon a placenta of adequate functional capabilities. The observations suggest that the placenta is similarly dependent upon its association with a normal healthy fetus.     

Density Studies on Lupins: I. Flower Development

In an August-sown experiment the pattern of flower developmentwas followed for cv. Ultra (Lupinus albus L.) and cv. Unicrop(L. angustifolius L.) grown at low (10 plants m^–2) andhigh (93 and 83 plants m^–2, Ultra and Unicrop respectively)densities. Dry weight increase of flowers on the main-stem inflorescenceand first lateral below the main-stem were compared at differentfloral stages. Maximum flower weight was reached just priorto the open flower stage and remained constant or declined untila pod formed or abscission occurred. The time period betweenmaximum flower weight and pod formation or abscission was upto 10 days. Emergence of the inflorescence was earlier and thefirst flower of Ultra opened 10 days before Unicrop. Developmentof each terminal raceme (inflorescence) was acropetal, withpods having formed on lower flower nodes when terminal flowerswere still quite immature. Laterals forming the next generationof inflorescences grew from axillary leaf buds below an inflorescencewhile it was in full flower. Sources of competition from connectedreproductive and vegetative metabolic sinks are discussed. Lupinus spp., lupins, flower development, planting density     

Cell proliferation patterns during development of the equine placenta

Placentation involves considerable growth and reorganization of both maternal and fetal tissues. In this investigation, immunohistochemical localization of the proliferation marker Ki-67 antigen was used to monitor cell division during placentation in mares. Endometrial biopsies were obtained from eight mares between day 14 and day 26 of pregnancy and from eight anoestrous mares that had been treated with various combinations of progesterone and oestrogen. Samples of endometrium and fetal membranes were obtained from 19 mares carrying normal horse conceptuses between day 30 and day 250 of gestation and from three failing extraspecific donkey-in-horse pregnancies. Proliferation in the superficial strata of the endometrium was increased by day 18 of gestation and this effect could be mimicked by supplementing with oestradiol benzoate during the last 6 days of a prolonged period (18-36 days) of progesterone administration. Fetal chorionic girdle cells were proliferating vigorously at days 30-32 of gestation, but stopped dividing after they invaded the endometrium, while the trophoblast cells of the allantochorion showed an increase in mitotic activity after day 38. The luminal epithelium of the endometrium started to proliferate only after the primary villi of the true epitheliochorial placenta had been formed, and during days 58-70 this effect was seen only in the pregnant horn in which placentation was further advanced. During the second half of gestation, most of the mitotic activity was confined to the periphery of the microcotyledons which were still growing. In the donkey-in-horse pregnancies, proliferation rates of the maternal and fetal epithelial at day 70 of gestation were markedly reduced in areas of heavy endometrial lymphocyte infiltration and poor placentation. These results provide a basis for further studies on factors that influence invasive and non-invasive placentation.     

Studies on cytosolic guanylate cyclase from human placenta.

We have purified the soluble form of guanylate cyclase from human placenta greater than 2400-fold. The enzyme shared several characteristics with the enzyme purified from other sources including molecular mass and subunit composition, activation by divalent cations, inhibition by ATP and Michaelis constants. The enzyme, however, had a lower absorption maximum in the Soret region (417 +/- 1 nm) than the enzyme from other sources and was activated only one-fifth as much by nitric oxide as the bovine lung enzyme. It appears that the heme prosthetic group in the human placental enzyme may be hexa-coordinate and in the bovine lung enzyme the heme group may be penta-coordinate.     

Studies into equine electrocardiography and vectorcardiography: I. Cardiac electric forces and the dipole vector theory

Theoretical consideration has been given in two horses to the properties of the electric field created by the equine heart acting as a simple electric generator. The principles of the vectorial theory have been applied to test the validity of application of the dipole concept. The cardiac electric forces, althrough complex in the immediate region of the heart, appear at the body surface in a similar form to those arising from a relatively immobile, single equivalent dipole. The potential value of the technique of vectorcardiography in cardiological investigations is briefly discussed.     

Studies on human tRNA. I. The rapid, large scale isolation and partial fractionation of placenta and liver tRNA.

A procedure for the large scale isolation of mammalian tRNA has been applied to the isolation of several grams of human liver, human placenta, rabbit liver and rat liver tRNA. This procedure entails an initial grinding of the tissue in phenol-sodium acetate at acidic pH, followed by DEAE cellulose chromatography. Procedures are also described for analysis of the purified tRNA on the basis of size, using controlled pore glass bean columns. In addition, the acceptor activity of isolated tRNAs has been determined using both the heterologous and homologous synthetases. The chromatographic profile of individual isoaccepting species using BD cellulose chromatography is shown and the 3' terminal nucleoside content was also determined. The methods described now make it feasible for large scale studies of mammalian tRNA enabling us to better understand the relationships between the structure of mammalian tRNA and its many diversified functions.     

Studies on the renal sac of the ascidian Molgula manhattensis. I. Development of the renal sac

The renal sac of the ascidian family Molgulidae (Tunicata, phylum Chordata) has been thought to function as a kidney, yet its structure, contents and activities seem incompatible with current generalizations regarding excretory processes in marine animals. The development of the renal sac is described here as part of a general effort to reexamine the organ's role in Molgula manhattensis. Light microscopy of living animals and fixed material has shown the following: (1) The renal sac begins to sequester concretions before the heart starts beating and before feeding begins. Therefore, blood circulation by heartbeat is not necessary for production or transport of the initial concretions, whatever its effects may be on the renal sac in older individuals. Ingested food cannot provide the initial concretion material. (2) In laboratory-raised animals, concretions appear in the renal sac before “renal sac organisms” (fungus-like organisms seen in the renal sac of all field-collected adults) can be detected. Thus, at least some portion of the concretions can be produced by Molgula in the absence of renal sac organisms. (3) No openings have been detected in the renal sac at any stage of its development, nor is there any evidence that concretions are dissolved or transported out of the renal sac. (4) The development and morphology of the renal sac are consistent with the hypothesis that the organ is an epicardial derivative, except that the renal sac arises from post-pharyngeal (presumptive gut) endoderm, rather than pharyngeal endoderm.     

Stability of equine lysozyme. I. Thermal unfolding behaviour.

The thermal denaturation of Ca(2+)- and apo-forms of equine lysozyme was followed by using far and near UV circular dichroism and intrinsic fluorescence methods. The difference found between the temperature dependence of the ellipticity at 222 nm and 287 nm, which show two stages in the thermal transition, and those at 228 nm and 294 nm, which indicate only one stage over a wide range of temperatures reflects that different subdivisions of the protein molecule are characterized by a different stability, cooperativity and pathway of denaturation. The first transition, reflected in the increase of the ellipticity at 222 nm and 287 nm, coincides with the transition detected by fluorescence and occurs at 30-50 degrees C for the apo-form and at 50-60 degrees C for the Ca(2+)-form of lysozyme. It seems to correlate with the transfer of some tryptophan residues to a more hydrophobic environment and with a local rearrangement of the tertiary and secondary structures. The unfolding transition detected by the decrease of the ellipticity at all wavelengths occurs nearly in the same temperature region for the apo- and Ca(2+)-forms, i.e. 50-80 degrees C and 55-80 degrees C, respectively. The presence of a Ca(2+)-binding loop in equine lysozyme may be partly responsible for the drastic destabilization of its structure as a whole both in the presence but especially in the absence of Ca2+ in comparison with hen and human lysozymes.