The anatomy and histology of the reproductive tract of the male Babirusa (Babyrousa celebensis).

The anatomy and histology of the reproduction organs of the male Babirusa (Babyrousa celebensis) were studied in 21 specimens collected between birth and approximately 17 years of age. In adult animals the testes were located in a subanal position against the caudal surface of the thigh musculature. Average adult testis length was 68.9 ± 5.1 mm, width was 40.3 ± 5.7 mm, and depth was 47.9 ± 7.0 mm (N = 11). The average combined adult testicular weight was estimated to be 82.7 ± 23.2 g (N = 11). The testes of newborn Babyrousa had descended through the inguinal canal into the scrotum before birth. Adult seminiferous tubules had an average diameter of 199 ± 33.6 µm (N = 9) and were randomly distributed among a smaller volume of Leydig cells. Connective tissue was sparse. In one 13-month-old prepubertal animal the diameter of the seminiferous tubules was 85.3 ± 16.1 µm (N = 7). The spermatozoa was 42.2 ± 4.9 µm (N = 19) long and had a flat, paddle shaped head, 6.3 ± 0.6 µm (N = 50) long, 3.9 ± 0.5 µm (N = 47) wide, and a thickness of approximately 0.5 µm. An apical ridge along its front represented the acrosome. The two adult vesicular glands each had an irregular shape and were approximately 48.7 ± 7.4 mm long, 25.6 ± 4.3 mm wide, and 20.6 ± 8.7 mm deep (N = 6). The prostate, comprising a corpus and disseminate parts, lay ventral to the vesicular glands partly embedded in the dorsal wall of the urethra. The paired adult bulbourethral glands were approximately shaped like prolate (elongated) spheroids and had a length of 51.2 ± 14.2 mm, a width of 22.6 ± 4.5 mm, and a depth of 14.4 ± 4.5 mm (N = 7). The secretions from the bulbourethral glands drained into the urethral recess, which in adults measured approximately 10 to 14 mm in length and was located caudodorsal to a narrowing of the pelvic urethra. The penis was 330 ± 16 mm long and 8.2 ± 0.6 mm in diameter, and rotated approximately two and a half turns counterclockwise along its longitudinal axis toward its free end. The small prepucial diverticulum situated dorsocranial to the penis tip in adult and prepubertal Babyrousa, in adults measured 22.0 ± 1 mm in length and 17.5 ± 2.6 mm (N = 3) in width.

Anatomy and histology of the reproductive tract of the female Babirusa (Babyrousa celebensis).

The anatomy and histology of the female reproductive tract of the Indonesian wild pig Babyrousa celebensis was studied by means of reproductive tracts obtained from seven animals aged between two and 22 years of age. The ovary appeared to have the ability to ovulate up to four ova at one time. However, the combined ovarian output seemed to average 1.86 ova. Ovulation can take place at any time from puberty to old age (22 years). The opening to the uterine tube was indicated by a 'flower-like' array of tall, broad epithelial 'petals' arising from the luminal surface of the funnel. The mucosal surfaces of these structures were covered in a mixture of prominent ciliated cells and bulbous secretory cells. The uterine tube followed a tightly convoluted path to the tip of the uterine horn. The uterus was proportionately short. The anatomical construction of the uterus was similar to those of other suids in that the columnar endometrium was heavily folded, there was a rich supply of uterine glands in the lamina propria, and the uterus was provided with a good blood supply. The cervix was thick walled and had a spiral lumen.

Distinct factors in plasma of preeclamptic women increase endothelial nitric oxide or prostacyclin.

The pathogenesis of preeclampsia is proposed to be due to uncharacterized circulating factors that activate endothelial cells. Support for this hypothesis is provided by in vitro activation of endothelial cells by plasma from preeclamptic women, eg, increased nitric oxide and prostacyclin generation. We performed molecular sizing, lipid extraction, and lipoprotein fractionation of plasma from normal pregnant and preeclamptic women and determined the ability of these plasma fractions to increase nitric oxide or prostacyclin generation by endothelial cells. Fractions from plasma of preeclamptic women were consistently more active than fractions from normal pregnant women, although characterization was qualitatively similar. The factors stimulating nitric oxide and prostacyclin were different. The factor (or factors) stimulating nitric oxide generation was extractable by charcoal and present in lipid extracts and lipoprotein isolates with a molecular weight greater that 1.5 million daltons, which is characteristic of a lipoprotein or lipoprotein aggregate. By contrast, activity to stimulate prostacyclin persisted after charcoal stripping or lipoprotein removal, partitioned to the aqueous fraction, and had a molecular weight of approximately 50,000 D. Two distinct factors in the blood of preeclamptic women alter endothelial function in vitro. This information should guide the search for circulating factors contributing to the pathophysiology of preeclampsia.

Evidence of endothelial activation and endothelial activators in cord blood of infants of preeclamptic women.

OBJECTIVE: In preeclampsia markers of endothelial activation (e.g., increased cellular fibronectin and activities that alter in vitro endothelial function (e.g., stimulation of nitric oxide and prostacyclin generation) are increased in the maternal circulation. We tested preeclamptic infant blood for these markers and activities and correlated these findings with fetal growth. STUDY DESIGN: Plasma was obtained from 17 term nulliparcus preeclamptic and normal pregnant women and their infants and from 8 additional preeclamptic mother-baby pairs from earlier gestations. Plasma cellular fibronectin and production of nitric oxide and prostacyclin by cultured endothelial cells exposed to 2% plasma were measured. RESULTS: Cellular fibronectin was higher in maternal plasma of preeclamptic than nonpregnant women (6.1 +/- 0.29 vs 4.2 +/- 0.27 microgram/ml, p < 0.01), as were stimulated endothelial nitric oxide and prostacyclin production (nitric oxide 42.5 +/- 3.9 vs 26.9 +/- 2.3 nmol nitrite/microgram protein/24 hours, p < 0.05; prostacyclin 261.7 +/- 31.2 vs 151.9 +/- 18.7 pg prostaglandin F1 alpha/microgram protein/24 hours, p < 0.05). In the preeclamptic infants cellular fibronectin was also greater (3.3 +/- 0.15 vs 2.6 +/- 0.14 microgram/ml, p < 0.01), as was endothelial nitric oxide production in response to the plasma (24.4 +/- 1.1 vs 21.4 +/- 0.09 mumol/L nmol nitrite/microgram protein/24 hours, p < 0.05). Prostacyclin production was not significantly different. In preeclamptic infants across a wide gestational age there was no correlation of endothelial activation and fetal growth. CONCLUSIONS: Infants of women with preeclampsia may be affected by endothelial dysfunction, as well as reduced uteroplacental perfusion.

Production of laminin B2 chain protein and messenger RNA by a murine neutrophil precursor cell line, 32Dc13.

Laminin is a heterotrimeric glycoprotein that plays a central role in promoting neutrophil chemotaxis, motility, and attachment to basement membrane. Rabbit peritoneal exudate neutrophils stain positively for laminin, which is presumed to be of exogenous origin and bound to laminin receptors on the cell surface. We examined 32Dc13 cells, a murine neutrophil precursor cell line, by immunoprecipitation. Northern blot analysis, flow cytometry, and electron microscopy for the endogenous production of laminin. Our results demonstrate that 32Dc13 cells endogenously produce a laminin B2 chain protein and messenger RNA (mRNA) without producing any detectable A or B1 chain protein or mRNA. The B2 chain protein was not secreted by the cells; rather it could be detected on the cell surface after treatment of cells with neuraminidase. These findings suggest the possibility of a novel role for the laminin B2 chain in myeloid development and function.

Macrophage handling of a tolerogen and the role of IL 1 in tolerance induction in a helper T cell clone in vitro.

The human gamma-globulin (HGG)-specific helper T cell clone AB.7.D7 can reconstitute the plaque-forming cell response of HGG-primed B cells. Tolerance induction at the level of T cell help results from exposure of the AB.7.D7 cells to 10 micrograms monomeric HGG. The monokine IL 1 was found to interfere with tolerance induction in AB.7.D7 cells in a dose-dependent manner. Furthermore, interference with tolerance induction was dependent upon the T cells being presented with IL 1 at the same time as monomeric HGG, the tolerogen. IL 1 and monomeric HGG could not be demonstrated to interact to make nontolerogenic soluble aggregates, however. It was found that monomeric HGG was unable to stimulate the production of either membrane or secreted IL 1 by splenic macrophages and in addition was not degraded by peritoneal exudate cells. Heat-aggregated HGG, which is highly antigenic and nontolerogenic, is a good stimulus for IL 1 production and is processed by macrophages into peptides of varying sizes. These data are consistent with the suggestion that a tolerogenic signal results from T cell recognition of a nondegraded antigen in the absence of a signal from IL 1. It is possible, however, that small amounts of processed antigen, undetectable by us, are involved.

Characterization of a substance in rat serum which modulates prolactin release in vitro.

A prolactin (PRL) inhibitory substance(s) present in rat serum has previously been shown to inhibit the release of PRL from rat pituitary cells in vitro (Hymer and Signorella, 1981). In this study, addition of rat serum to rat pituitary cells cultured in serum-free medium inhibited the release of PRL in dose-dependent manner. Neither the form of the released PRL molecule, nor its biological or immunological activity, was altered by rat serum treatment. Serum prepared from clotted blood previously incubated at 37 degrees C always contained more PRL inhibitory activity than serum from blood incubated at 4 degrees C; whole blood serum, but not plasma-derived serum, contained inhibitory activity. The inhibitor therefore comes from a formed element in the blood. Results of several experiments indicated that buffy coat cells, adherent blood cells and peritoneal cells released the inhibitor, but platelets and RBCs did not; it is suggested that a macrophage/monocyte is probably the cellular source of the PRL inhibitor. Gel permeation high-performance liquid chromatography (HPLC) of rat serum indicated that the inhibitory activity was associated with high molecular weight material, i.e. 115,000-215,000. The PRL inhibitory substance was trypsin sensitive, not extractable with butanol/diisopropyl ether, precipitated at 40% saturated ammonium sulfate, associated with material having an isoelectric point of 4.96 +/- 0.26, and not adsorbed with protein-A. These characteristics suggest the proteinaceous nature of the inhibitory substance. The inhibitor did not suppress PRL release until 9 h after addition to cultured cells, but decreased PRL synthesis was evident within 4 h of treatment. The inability of the dopamine antagonist, haloperidol, to block the inhibitory effect of rat serum indicated that the material does not exert its effect through the dopamine receptor.

An enzyme-linked immunosorbent assay for rat prolactin.

A sensitive and specific competitive enzyme-linked immunosorbent assay (ELISA) for rat prolactin was developed using reagents from the National Institute of Arthritis, Diabetes, Digestive Diseases and Kidney. In this assay soluble prolactin and prolactin adsorbed to a solid-phase support compete for rabbit anti-prolactin antibody binding sites. Therefore, a high concentration of soluble prolactin in the sample will result in a low concentration of antibody immobilized to the adsorbed prolactin. The immobilized antibody-prolactin complex is detected and quantified using goat anti-rabbit immunoglobulin G covalently conjugated to the enzyme horseradish peroxidase. Assay parameters were optimized by investigating the concentration of reagents and the reaction kinetics in each of the assay steps. The assay can be performed in 24 h. A sensitivity range of 0.06 to 6 ng in the region of 90 to 10% binding was obtained. Near 50% binding (0.6 ng), the intraassay coefficient of variation (CV) was 4.2% and the interassay CV was 7.6%. The correlation between radioimmunoassay and the ELISA was 0.868. Selected applications of the assay are described. The assay should prove a useful alternative to the radioimmunoassay in those instances where steps involving the use of 125I become limiting, for example, iodination facility and gamma counter availability or prolonged reagent storage.

Rat serum inhibits prolactin release from rat pituitary mammotrophs in culture.

Rat pituitary cells maintained in culture for 3 days in alpha-MEM supplemented with 17% horse serum produce and release 4-8X the quantity of prolactin (PRL) originally contained in the cells at the time of plating. Addition of small (200 microliter) quantities of rat serum repeatedly suppressed PRL release by approximately 50%. Inhibition was related to dose of rat serum. Other studies showed (a) that the cells could recover from serum-induced inhibition, (b) that the effect was independent of the age of the pituitary and/or serum donor, (c) that GH release was unaffected by rat serum addition, (d) that the effect could not be attributed to proteases in the serum preparations and (e) that addition of T3 to the culture medium prevented the inhibitory response. The inhibitory material(s) in rat serum is stable on freezing; is non-dialyzable; is heat-labile; is stable over 3 days incubation at 37 degrees C; and is associated with molecules between 85 000 and 146 000 dalton. Inhibitory activity was generated upon incubating whole clotted blood at 37 degrees C for 23 h. It is suggested that the inhibitory material might (a) be released from platelets during clotting or (b) be generated from an inactive precursor molecule cleaved by proteases released during the clotting process.

Feedback effects of mammary gland tumors on the host pituitary prolactin cell.

Prolactin (PRL) cells in pituitary glands of F-344 female rats bearing transplantable mammary gland adenocarcinomas (13762, 3230) were smaller, contained less intracellular hormone, and released less hormone in culture relative to non tumor-bearing littermates. In some instances removal of the tumor by surgery or chemotherapy (PAM) resulted in restoration of function in vitro. On the other hand, PRL cells from pituitaries of breast cancer patients were often hypertrophied. Since these patients had mastectomies prior to hypophysectomy, the data are consistent with the hypothesis that the tumor exerts negative feedback at the pituitary or hypothalamic level. In support of this hypothesis we have discovered a potent PIF in rat serum. In addition, an experimental protocol involving a) encapsulation of pituitary cells in Amicon hollow fibers, b) their implantation in ectopic sites and c) their subsequent in vitro culture yields data which suggest that the brain of the tumor bearing rat suppresses PRL release. It is suggested that decreased PRL cell function in animals bearing certain mammary tumors ultimately favors metastatic activity of the tumor cell.