A functional analysis and the potential clinical significance of 7 categories of sperm morphology.

Sperm morphology is a critical factor in the evaluation of infertile men. Patients often exhibit a morphology profile with consistent increases of specific abnormal categories of sperm morphology. We analyzed the functional ability of 7 categories of sperm morphology using assays that measure sperm viability longevity, deoxyribonucleic acid packing, acrosomal status, and the stimulation of the acrosome reaction with human cumulus/oocyte complexes and calcium ionophore A23187. Each morphology category was separately analyzed initially and at 3, 6, 12, 18 and 24 hours of in vitro culture. The data indicate that all categories of abnormal sperm morphology exhibit various functional debilities. However, tapered sperm most closely approached normal sperm function. All abnormal categories possessed subpopulations of sperm that functioned normally to the limits of these assays. These data indicate that increases in any of the categories of abnormal sperm morphology may lead to decreased fertility due to diminished functional ability indicated by the assays performed in this study.

Pregnancies following implantation of an artificial fallopian tube in rabbits.

The first cases of pregnancies obtained in vivo in animals after implantation of a Teflon artificial fallopian tube model are presented. The Teflon prosthesis was implanted into the uterine cornua of rabbits and fixed subcutaneously in the abdominal wall. Five to eight blastocysts obtained from donor rabbits were transferred through the tubing into each recipient animal's uterus 2 to 14 days after placement of the device. On posttransfer day 10, normally developed pregnancies were visible and confirmed histologically in 4 of 7 animals.

Metabolism of progesterone by avian granulosa cells in culture.

Previous studies have demonstrated that progesterone is the primary product of steroidogenesis in avian granulosa cells during short-term incubation. However, during more prolonged culture, lasting several days, the progesterone content in the medium was found to decrease progressively, indicating in vitro metabolic conversion. In the present study we have isolated and identified a number of progesterone metabolites. Granulosa cells, isolated from mature ovarian follicles of laying hens, were cultured in medium 199 supplemented with fetal calf serum and containing [14C]progesterone. After 4 days in culture, cells + media were extracted and the radioactive metabolites separated and identified by TLC, HPLC and GC-MS. Several of the metabolites were further characterized by derivatization and crystallization to constant specific activity. A total of 24 radioactive substances was detected. Of these, 15 have been positively identified, 5 tentatively and the remaining 4 are unidentified. The principal metabolite, representing more than 45% of the total radioactivity, was identified as 3 alpha-hydroxy-5 beta-pregnan-20-one. In addition, significant amounts of 3 alpha-hydroxy-5 alpha-pregnan-20-one (5.76%), 5 beta-pregnane-3,20-dione (3.05%), and 5 alpha-pregnane-3,20-dione (2.95%) were detected and identified. The results indicate that avian granulosa cells possess 3 alpha-hydroxy-steroid dehydrogenase (3 alpha-HSD), 17 beta-HSD, 20 alpha-HSD, 20 beta-HSD, 17 alpha-hydroxylase, C17-20-lyase and 5 alpha- and 5 beta-reductase activities. These enzyme activities may convert progesterone to biologically inactive or less active metabolites. However, a functional role for some of these metabolites cannot be ruled out.

Errant percutaneous Greenfield filter placement into the retroperitoneum.

Percutaneous Greenfield filter placement through a femoral vein approach resulted in errant retroperitoneal placement in a patient with pulmonary emboli. A properly positioned filter was then placed surgically through the right internal jugular vein. The patient subsequently required removal of the errant retroperitoneal filter because of genitofemoral nerve irritation. Problems associated with percutaneous Greenfield filter placement and pertinent literature are discussed.

The effects of ionophores on steroidogenesis and morphology of avian granulosa cells.

Granulosa cells, isolated by collagenase digestion from the mature ovarian follicle of laying hens, were incubated in the presence of two ionophores, lasalocid (X537A) and ionomycin, to determine their effects on basal and stimulated steroidogenesis, as well as their effects on various cell parameters including DNA, RNA, and protein synthesis. Both ionophores caused a dose-dependent inhibition of agonist-promoted progesterone production and, in the presence of calcium, a small but significant increase in basal output of progesterone. Whereas the conversion of pregnenolone to progesterone was unaffected by the ionophores, the activity of cholesterol side-chain cleavage enzyme was inhibited in a dose-related manner. Both ionophores decreased cellular levels of ATP and inhibited the incorporation of radioactively-labeled precursors into DNA, RNA, and proteins. Morphologically, ionophore-treated cells showed swelling of the rough endoplasmic reticulum. Similar morphological changes were also observed in cells treated with oligomycin, a known metabolic inhibitor. These results suggest that the ionophores lasalocid and ionomycin impair release of energy and thereby exert the principal cause of the inhibited steroidogenic response by granulosa cells to a variety of agonists.