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Biomedical subjects

J Halme

Publications and source records attributed to J Halme.

At least 55 records · Page 3Linked to original sources

Artificial donor insemination: analysis of 149 cases at North Carolina Memorial Hospital.

We analyzed 149 cases of artificial insemination with donor semen at North Carolina Memorial Hospital, Chapel Hill, to study its effectiveness. An overall pregnancy rate of 62.1% was achieved. Based on the mean monthly fecundity rate, the average monthly pregnancy rate was 16.5%, and the cumulative pregnancy rate after three, six, and ten cycles was, respectively, 40.3%, 68.5%, and 95.5%. A uniform evaluation of the women and correction of any treatable female factors before insemination contributed to a high success rate. Diagnostic laparoscopy after three to four failed insemination cycles yielded a high percentage of abnormal findings. These data suggest that almost all properly screened patients will conceive if allowed to undergo a sufficient number of inseminated cycles and that this procedure is a highly effective alternative means of circumventing male infertility.

Actuarial Analysis↗

Retrograde menstruation in healthy women and in patients with endometriosis.

Blood was found in the peritoneal fluid in 90% of women with patent tubes at laparoscopy during perimenstrual time. If the fallopian tubes were occluded, then only 15% of patients had evidence of blood in the pelvis. Also, 90% of patients with endometriosis and eight of nine women on oral contraceptives had bloody fluid during the menstrual period. The present observations indicate that retrograde menstruation through the fallopian tubes into the peritoneal cavity is a very common physiologic event in all menstruating women with patent tubes.

Ascitic Fluid↗

Increased activation of pelvic macrophages in infertile women with mild endometriosis.

Pelvic fluid was collected from 66 women undergoing laparoscopic sterilization or diagnostic laparoscopy for evaluation of infertility. Cells consisting mainly of macrophages were separated, counted, and subjected to histochemical staining for acid phosphatase and myeloperoxidase as markers of cell irritation. Pelvic fluid was analyzed for acid phosphatase, neutral protease, and extractable prostaglandin E2 and F2 alpha. A higher proportion (46% versus 15%) of the macrophages in the group with mild endometriosis exhibited positive staining for acid phosphatase as compared with the fertile group. Pelvic fluid from patients with mild endometriosis had higher acid phosphatase and neutral protease activity than that from fertile patients (p less than 0.05, p less than 0.01). The content of either prostaglandin was not significantly higher in the endometriosis group as compared with the fertile group. The results suggest that mild endometriosis is associated with activation of macrophages and release of active substances into peritoneal fluid that may be responsible for the associated infertility.

Acid Phosphatase↗

Purification and properties of rat uterine procollagenase.

A procollagenase from monolayer cultures of postpartum rat uterine cells has been purified. The crucial step in the purification is the binding of the procollagenase from crude, fetal bovine serum-containing culture medium to heparin-Sepharose, followed by elution with extremely low concentrations (5-10 nM) of dextran sulfate. Resultant eluates contain 8-10% procollagenase. Purification is completed by ion-exchange chromatography on DEAE-Sepharose, gel filtration on AcA-44, and chromatography on blue-Sepharose. Rat uterine procollagenase appears as a protein doublet of Mr approximately 58,000, as indicated by two polyacrylamide gel electrophoresis systems, by AcA-44 chromatography, and by equilibrium sedimentation ultracentrifugal analysis. The proenzyme forms are converted by trypsin to an active enzyme doublet of Mr approximately 48,000. Small amounts of active enzyme, which are often generated during the purification, are electrophoretically indistinguishable from trypsin-activated collagenase. Active collagenase can be separated from the zymogen forms by DEAE-Sepharose chromatography. The two forms of the proenzyme doublet can be partially separated by gel filtration on AcA-44 and preliminary analysis indicates each has equal collagenolytic activity. The amino acid analysis of rat uterine collagenase reveals it to be markedly different from two other vertebrate collagenases whose composition is known. The uterine proenzyme is unusually rich in glycine and in the hydroxy amino acids and is considerably more acidic than the human skin fibroblast collagenase, consistent with the different ion-exchange behavior of the two molecules. The specific activity of rat uterine collagenase at 37 degrees C is approximately 3000 micrograms collagen/min/mg, using native reconstituted guinea pig skin type I collagen fibrils as substrate. The enzyme cleaves denatured collagen, but fails to attack a variety of noncollagen proteins.

Animals↗

Heterogeneity of human peritoneal macrophages: cytochemical and flow cytometric studies.

Human peritoneal macrophages obtained from females undergoing laparoscopic examination have been investigated. Morphologic heterogeneity as well as a wide variation in the content of nonspecific esterase, acid phosphatase (AP), and myeloperoxidase (MP) were observed with standard cytochemical stains. This heterogeneity was evident within the same sample as well as between different individuals. Phagocytic activity was also highly variable. In only 3 cases out of 85 samples screened could human peritoneal macrophages be characterized as resident (peroxidase negative, acid phosphatase negative) by the criteria used in rodent systems, thus indicating that female peritoneal macrophages are continuously responding to stimuli/irritation. A newly developed flow cytofluorometric method was used to quantitatively document the macrophage heterogeneity in enzyme expression and phagocytic activity. Population distributions derived from single-cell analysis of various enzyme activities and phagocytic levels further emphasized the spread in expression of these parameters both within the individual peritoneal samples as well as between donors. Simultaneous determination of two markers such as leucine aminopeptidase (LAP) and zymosan ingestion indicated that the association between these features varied markedly between individuals. It is hoped that the use of this multiparameter flow cytometric technique on in vitro differentiating blood monocytes, subjected to different stimuli, will give us a better understanding not only of the development of monocytes to macrophages but also of the sources of heterogeneity in normal human resident macrophages.

Acid Phosphatase↗

Pelvic macrophages in normal and infertile women: the role of patent tubes.

The volume of peritoneal fluid and its macrophage content were examined in 80 women undergoing laparoscopy. The amount of pelvic fluid was not dependent on the patency of the fallopian tubes, and no statistical difference in the fluid volume was observed between the infertile groups, including those with endometriosis, as compared to the fertile group. The number of pelvic macrophages in fertile women was extremely high during the menstrual period, and during the rest of the cycle, it remained at a basal level. The number of pelvic macrophages in women with occluded tubes was low. Four of 21 infertile patients with mild endometriosis had very high numbers of pelvic macrophages, even in the luteal phase. These results suggest that the passage of endometrial irritants through the tubes elicits a macrophage response in the peritoneal cavities of both normal women and patients with endometriosis. Although most patients with endometriosis had normal numbers of peritoneal macrophages, it is possible that these macrophages differ qualitatively from normal, and this could explain the associated infertility.

Ascitic Fluid↗

Effect of pelvic fluid from endometriosis patients on human sperm penetration of zona-free hamster ova.

Peritoneal fluid (PF) samples from fertile women, infertile women without signs of endometriosis, and infertile women with endometriosis were examined for effects on motility of normal spermatozoa and for effects on their fertilizing capacity as measured by the interspecies penetration test (ISPT). All unheated peritoneal fluid samples caused rapid destructive changes in the female hamster gametes, presumably due to the presence of unidentified serum components, and caused clumping of spermatozoa but no change in sperm motility. Heat-treated fluid samples were therefore used in subsequent experiments to assess their effect on the ISPT. The mean penetration rate for fertile women was 58%; for infertile women without endometriosis, 55%; and 54% for infertile women with endometriosis. The differences between the penetration rates in these patient groups were not significant. No adverse thus found on the fertilizing capacity of fertile spermatozoa.

Animals↗

The removal of fallopian tubes has no adverse effect on subsequent ovarian function in rabbits.

The effect of bilateral salpingectomy on ovarian function was studied in the rabbit. Nine rabbits underwent a microsurgical removal of both fallopian tubes, and nine other rabbits underwent sham operations. Four weeks after surgery the rabbits were mated, and the number of ovulation sites were recorded 18 hours after mating. Serum progesterone (P) levels were also determined prior to the operation and at 6 days and 8 days after mating. Eight days after mating the ovaries were recovered, and the corpora lutea were removed and weighted. The mean number and weights of corpora lutea as well as serum P levels did not differ significantly between the groups. These findings suggest that the absence of fallopian tubes has no effect on subsequent ovulatory function in this animal model.

Animals↗

Gonadotropin receptors of human corpus luteum during menstrual cycle and pregnancy.

Specific high-affinity low-capacity binding of human chorionic gonadotropin (hCG) and lutropin (hLH) was demonstrated in the plasma membrane fractions of human corpora lutea. The number of binding sites for both hormones increased from early to late luteal phase, whereas regressing corpus luteum from proliferative phase did not bind either hormone. On the basis of apparent dissociation constants the affinity of the receptor for hLH is highest during early luteal phase and decreases toward the end of the cycle, which may reflect an increasing insensitivity of the corpus luteum to circulating hLH. By contrast, the affinity of the receptor for hCG is highest in the midluteal phase. There are gonadotropin binding sites in the human corpus luteum also during pregnancy, but they are saturated by endogenous hCG. Evidence for this was obtained by elution of hCG with 0.15M sodium chloride at pH 2.3 from washed plasma membrane fractions of luteal tissue from six to 16 week's gestation. After acid treatment and neutralization these preparations showed specific binding for 125I-labeled hCG, but not for 125I-labeled hLH. Our results demonstrate a shift in the balance of affinity of the gonadotropin receptor from hLH to hCG during the course of luteal phase, and during pregnancy the binding sites appear to be available for hCG only.

Cell Membrane↗

Extraction of collagenase from the involuting rat uterus.

Collagenase (EC 3.4.24.3) activity can be measured directly in homogenates of the involuting rat uterus. Latent forms of collagenase are activated by a brief exposure to trypsin; trypsin activity is then blocked with soybean trypsin inhibitor. Homogenizing conditions have been developed that permit 90-95% recovery of the total active and latent collagenase activity in a 6000 X g pellet, where it is presumably bound to its collagen substrate. This insoluble activity can then be extracted by heating to 60 degrees C for 4 min in 0.04 M Tris - HCl buffer, pH 7.5, containing 0.1 M CaCl2. Methods are presented for the estimation of the recovery of collagenase in the extracts; this approximates 65-70% of the total. Small amounts of activity can also be extracted from rat liver and kidney. This extraction procedure should be of use in purifying collagenase without culturing the enzyme-producing tissue and in the direct assay of tissue collagenase activity. The activity extracted from rat uterus has been proven to be collagenase by its characteristic pattern of collagen breakdown products on disc electrophoresis and by the split of tropocollagen at interband 41 as shown by electron microscopy of reconstituted fragments. The activity is inhibited by EDTA, and this inhibition is not reversed by calcium or zinc ions.

Animals↗

Effect of progesterone on collagen breakdown and tissue collagenolytic activity in the involuting rat uterus.

Progestational agents were studied for their effects on collagenolytic activity and loss of collagen and wet weight from the involuting post-partum rat uterus. Administration of very large doses of progesterone (80-150 mg/day) significantly retarded uterine involution and loss of collagen. This was accompanied by a significant reduction in uterine collagenolytic activity. By 72 h post partum, uteri of rats treated with 150 mg progesterone/day had wet weights 30% above, collagen 85% above, and collagenolytic activity 45% below, those of the control uteri. Similar effects were produced by 17alpha-acetoxy-6alpha-methylprogesterone at the same dosage levels. However, the progestational agent 6-chloro-17alpha-acetoxypregna-4,6-diene-3,20-dione acetate had no effect in this system.

Animals↗