Ove Nilsson

Enzyme activities and ultrastructure of a membrane fraction from human erythrocytes

Abstract The homogeneity of an isolated membrane fraction of human erythrocytes was studied by density gradient centrifugation in silica solution. A main component (about 90% on a nitrogen basis) was obtained as a single band at a density of 1.08. When analyzed by electron microscopy, this component was found to consist of a membranous structure which, in contrast to isolated ghosts, appeared to be essentially devoid of cytoplasmic structures. The main component, obtained at a density of 1.08 of the gradient, was shown to contain glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase and adenylte kinase. Thus, the present results strongly support the view that part of the glyceraldehyde-3-phosphate dehydrogenase, phophoglycerate kinase and adenylate kinase of the human erythrocyte is intimately associated with the membranous structure.

Prostasome-like granules from the PC-3 prostate cancer cell line increase the motility of washed human spermatozoa and adhere to the sperm

OBJECTIVE Prostasome-like granules are present in the PC-3 prostate cancer cells. Since the seminal prostasomes are able to promote the forward motility of human spermatozoa, we conducted a study to determine whether PC-3 prostasomes exerted effects similar to those of seminal prostasomes on buffer-washed spermatozoa from normospermic semen samples. STUDY DESIGN We used computer-assisted sperm analysis (CASA) and immunostaining of prostasomes to find out where these granules are located on the spermatozoa. RESULTS Addition of PC-3 prostasomes increased the proportion of motile spermatozoa from 12-15% to 50-70% (p<0.001). The optimal protein concentration of these prostasomes was 0.1mg/ml. Heat treatment of PC-3 prostasomes did not decrease their motility-promoting effect. Immunostaining with anti-prostasome monoclonal antibody (mAb78) revealed that the PC-3 prostasomes and seminal prostasomes adhered to the sperm cells. The staining, which occurred all over the spermatozoa, was intense on the mid-pieces and weaker on the sperm heads. Herewith, some prostasome component may activate the spermatic mitochondrial function, thus increasing sperm motility. CONCLUSION It is concluded that PC-3 prostasomes bear a functional resemblance to seminal prostasomes as regards sperm motility promotion.

Endometrial effects of levonorgestrel and estradiol: A scanning electron microscopic study of the luminal epithelium

Fertile women in the follicular phase possessed an uterine luminal surface with many ciliated cells and with non-ciliated cells, which carried numerous, fairly long microvilli. A moderate number of the non-ciliated cells had an irregular surface with small apical protrusions. Postmenopausal women had an endometrial surface containing rather flat cells. Ciliated cells were seldon encountered, and the non-ciliated cells possessed mostly only few short microvilli. When menopausal women had been wearing estradiol-containing intravaginal rings for three weeks, the uterine surface had developed many ciliated cells, and the non-ciliated cells now possessed many long microvilli. This appearance resembles that appearing during the follicular phase. Fertile women with levonorgestrel-containing subdermal implants or intravaginal rings showed a surface epithelium with few ciliated cells and with non-ciliated cells possessing short and irregular microvilli; that is, an epithelium less developed than that from a cyclic women. Adding estradiol to the levonorgestrel-containing intravaginal rings resulted in an estrogen response with an increase in number and length of the microvilli and an appearance of a few small apical protrusions.

Blastocyst surface charge and implantation in the mouse

Abstract During delay of implantation, the mouse blastocyst possesses several negatively charged sites on its surface and it is surrounded closely by the uterine epithelium. After activation for implantation by an injection of estrogen, the surface charge of the blastocyst remains unchanged for 8 hrs after the injection, although the blastocyst now lies free in the uterine secretion. Fourteen hrs after the injection, the surface charge is slightly less negative,but at 20 hrs the number of negative sites is markedly reduced. At this stage the blastocyst has resumed close contact with the uterine surface and has begun to invade the endometrium. It is concluded that the blastocyst can form close contact with the uterine surface both when inactive and possessing many negative surface sites and also when invasive and possessing few negative sites. The change in surface charge, which precedes blastocyst attachment, seems to occur simultaneously with a decrease in trophoblast antigenicity. Attention is drawn to the similarity in changes for the trophoblast at implantation to those of cultured cells at transformation.

Ultrastructure of human uterine epithelium at the time of implantation after postovulatory administration of norethindrone.

The impact of postovulatory norethindrone treatment upon the uterine epithelium of normal women at the time of implantation was studied in 7 women for a total of 11 menstrual cycles (3 cycles served as controls). A total of 150-300 mg norethindrone was administered orally for 2-4 days postovulation. Plasma progesterone, estradiol, and norethindrone levels were determined daily and an endometrial biopsy was obtained at the expected time of implantation of each cycle for light- and electron-microscopy studies. No consistent differences were revealed by light microscopy, however, electron microscopy revealed larger mitochondria and the presence of nucleolar channel systems after treatment, suggesting an increased progesteronelike influence in spite of the decrease in progesterone plasma levels. It suggested that these structural changes might change the functional properties and thereby impair normal implantation.

Ultrastructural Changes of Erythrocyte Membranes Isolated with Colloidal Silica Solution (Ludox)

The surface morphology of membrane fractions of human erythrocytes isolated by gradient centrifugation with a colloidal silica solution (Ludox) differed from the structure of control membranes isolated without the silica solution. The altered surface had a wrinkled appearance with a granular texture, compared with the smooth appearance of the controls. Thus the colloidal silica solution changed the ultrastructure of erythrocyte membranes. Possible mechanisms for the influence of the silica particles are suggested: secondary chemical bonds, e.g. hydrogen bonds, might well have been established between the silica particles and components of the erythrocyte membrane like spectrin of the inner surface of the membrane. Less probable is an attachment to positive sites of the outer side of the membranes as well as any association to the pitlike infoldings of the erythrocyte surface.

The Amount of Zinc Detected in Washed Human Spermatozoa

Samples of human semen, the supernatants from washings of these samples, and the suspensions of washed spermatozoa, were examined for zinc in a Perkin-Elmer spectrophotometer. Almost all the zinc was found in the original semen samples and in the first washings. Zinc could not be detected in the heads of fourfold-washed spermatozoa on analysis in a wavelength dispersive X-ray spectrophotometer attached to a scanning electron microscope.

Some clinical results of in vitro fertilization by the Malmö group, Sweden

In our in vitro fertilization program, only clomiphene/human chorionic gonadotropin-stimulated cycles have been used, yielding 1.6 preovulatory oocytes/woman at follicle aspiration. The cleavage rate was significantly influenced by the percentage of abnormal spermatozoa and by the percentage of motile spermatozoa in the spermiograms. At embryo replacement, a tilted uterus obtained by a full bladder in the woman simplifies the replacement procedure. In the last 12 months, 47 embryo transfers have been carried out, resulting in five clinical pregnancies. Of these pregnancies, one has proceeded to term, one resulted in a miscarriage, and three resulted in ectopic pregnancies.

Scanning electron microscopy of human preimplantation endometrium in normal and clomiphene/human chorionic gonadotropin-stimulated cycles**Supported by the Swedish Medical Research Council (Project 12X-70) and the “Expressen” Prenatal Research Foundation.

Embryo transfer after in vitro fertilization of oocytes obtained from unstimulated or stimulated women is usually carried out earlier than when an embryo developed in vivo reaches the womb. The possible asynchrony between the developmental stage of the embryo and that of the endometrium at transfer might reduce the prospects for implantation; so also might any secondary effect of stimulation. Preimplantation endometrium dated by plasma hormone analyses of 17 beta-estradiol and progesterone in five unstimulated and eight stimulated women was surveyed. Three features were examined, namely, the frequency, distribution, and appearance of ciliated cells, nonciliated cells, and apical protrusion. Wide regional variations were observed, but no continuous or consistent changes or differences were found from ovulation to the time for implantation in unstimulated and stimulated cycles. Therefore, from the morphologic point of view, the surface might be ready to accept a transferred embryo at any time within a range of days after ovulation.

Changes in Fertility and Morphology of Sensitized Rat Uterine Endometrium upon an Intra-uterine Injection of Antigen

To correlate fertility and endometrial morphology during an immune reaction, pregnant rats previously immunized with BCG vaccine were injected with .05 ml BCG vaccine in 1 uterine horn on Day 3 of pregnancy and the number of fetuses counted during Days 10-16. Spayed animals were given progesterone for 4 days and then an injection of BCG serum or vaccine in 1 uterine horn. The uteri were studied microscopically at intervals after injection. An intrauterine injection of BCG vaccine completely inhibited implantation. An intrauterine injection of serum to immunized or nonimmunized animals did not block implantation but seemed to lower fertility. Electron microscopy of the uteri of vaccinated spayed progesterone-treated animals showed an extensive development of the Golgi apparatus and granular endoplasmic reticulum. Large vacuoles were present, but there was a disappearance of the apical vesicles which are characteristic for a normal preimplantation stage in the rat. In the immediate type of hypersensitivity, the uterine epithelium is insufficiently changed to inhibit implantation.