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Antibodies against spermatozoa in infertile women with poorly invading spermatozoa on postcoital tests.

Sixty-four women with poorly invading spermatozoa on postcoital tests (PCT's) were investigated for serum spermatozoal antibodies by the immunofluorescent technique. Control groups tested were women with explained infertility but normal PCT's, normal pregnant women, and normal previously pregnant women. A positive immunofluorescent test was obtained in 19 of 64 women with abnormal PCT's compared with 26 of 51 patients with explained infertility, including 11 of 13 who had well-identified gynecological disorders, 12 of 61 pregnant women, and one of 59 previously pregnant normal women. High titers (larger than or equal to 1/200) were found in a few patients in all groups except normal nonpregnant women. In no patient was evidence of complement fixation found. The immunofluorescent test, as carried out here, is therefore of little value in discriminating a group of patients in whom immunologic abnormalities are the primary cause of the infertility.

Adolescent↗

A healthy birth after intracytoplasmic sperm injection by using immotile testicular spermatozoa in a case with totally immotile ejaculated spermatozoa before and after Percoll gradients.

Total asthenozoospermia is a severe problem, as only a micromanipulation technique can assist the couple, and even then fertilization and pregnancy rates are very low. The first healthy birth, achieved by using testicular immotile spermatozoa in a case with total asthenozoospermia before and after Percoll gradient preparation, is reported.

Adult↗

Timing of oocyte activation, pronucleus formation and cleavage in humans after intracytoplasmic sperm injection (ICSI) with testicular spermatozoa and after ICSI or in-vitro fertilization on sibling oocytes with ejaculated spermatozoa.

In the first study, we evaluated 101 oocytes [2, 4, 6, 8, 16, 18 and 20 h after intracytoplasmic sperm injection (ICSI)] that had been microinjected with testicular spermatozoa. Of the 70 normally fertilized oocytes (69%) 30 (43%) had two pronuclei by 6 h after ICSI. Fifty-one (73%) by 8 h, 69 (99%) by 16 h and four of them by 20 h cleaved to the 2-cell stage. In the second study, 95 cumulus-corona-oocyte complexes (CCOC) were divided into two groups. Forty-seven CCOC were inseminated by conventional in-vitro fertilization (IVF) and 40 metaphase-II oocytes by ICSI. Oocytes were evaluated at 2, 4, 6 (only after ICSI), 8, 10, 12, 18, 20, 22, 24, 26, 28 and 30 h after both ICSI and IVF. After IVF, 35 oocytes were fertilized normally (75%), four of which (11%) had two pronuclei by 8 h, 11 (31%) by 10 h, 27 (77%) by 12 h and 35 (100%) by 14 h. The first cleavages had occurred by 24 h after insemination (four oocytes, 11%). After ICSI, 34 oocytes were fertilized normally (79%), 13 of which (38%) had two pronuclei by 6 h, 27 (79%) by 8 h and 32 (94%) by 10 h. Three oocytes cleaved by 20 h after microinjection (9%) and 19 by 24 h (56%). Pronuclei developed asynchronously in six oocytes after ICSI (18%) as opposed to 16 oocytes after IVF (46%). The results of this study suggest that the timing of pronuclear formation is no different when a testicular spermatozoon is microinjected into the oocytes from when an ejaculated spermatozoon is injected. Secondly, pronuclear development and first cleavage generally take place 4 h sooner after ICSI than after IVF. On the other hand, a higher proportion of oocytes develop two pronuclei asynchronously after IVF than after ICSI.

Adult↗

'9 + 0' axoneme in spermatozoa and some nasal cilia of a patient with totally immotile spermatozoa associated with thickened sheath and short midpiece.

A patient is described with 100% immotile spermatozoa. The sperm cells lack the central pair of microtubules or the complete axoneme. This defect is associated with a thickened fibrous sheath and a shortened midpiece containing defective mitochondria. Fifteen per cent of the nasal cilia of this patient lack the central pair of microtubules. Eighteen similar cases have been described in the literature suggesting that this association of ultrastructural defects may be considered a syndrome.

Adult↗

Membrane glycoproteins from spermatozoa: partial characterization of an integral Mr = approximately 24,000 molecule from rat spermatozoa that is glycosylated during epididymal maturation.

Spermatozoa from the rat cauda epididymidis were treated with either the galactose oxidase-NaB[3H]4 or the NaIO4-NaB [3H]4 technique to label cell surface moieties of galactose and sialic acid, respectively. Following extraction with 40 mM octyl-beta-D-glucopyranoside (OBG), electrophoresis in sodium dodecylsulfate-polyacrylamide gels (SDS-PAGE) revealed a single radioactive peak migrating at Mr = approximately 24,000 in 11.2%, 14% and 16.8% tube gels. SDS-PAGE of the same OBG extract on 5.6% and 14% gels showed that this molecule was the same as that reported elsewhere, having a molecular weight varying from 32,000 to 37,000. The amount of labeled molecule extracted with 8 M urea or with 6 M guanidine-HCl was 30 and 50%, respectively, of that achieved with OBG. However, when labeled sperm were treated under other conditions (at pH 8, pH 3, 0.1-3 M NaCl [at pH 7.2], 5 mM ethylenediaminetetraacetic acid [EDTA], 20-200 mM dithiothreitol or 20-200 mM betamercaptoethanol, at varying temperatures and extraction times), the amount of labeled molecule extracted was less than 1% of that obtained with OBG. The molecule aggregates in aqueous buffers, as shown by chromatography using Sephadex G-100 and by centrifugation through a sucrose density gradient. Analysis by charge-shift electrophoresis suggested that the molecule contains an exposed hydrophobic domain(s). Mild trypsin treatment released all the labeled carbohydrate with the majority of the label attached to a Mr = 10,000 fragment. These data support the hypothesis that the molecule is an integral membrane glycoprotein, and suggest that it is only partially buried in the lipid matrix of the plasma membrane.

Animals↗

Use of spermatozoa for artificial insemination. I. Fertilizing capacity of fresh and frozen spermatozoa in sows on 36 farms.

A field trial was conducted on 36 farms in the Netherlands to compare the fertilizing capacity of fresh and frozen-thawed boar spermatozoa. Four-hundred and fifty-one sows were artificially inseminated with semen than that had been frozen and thawed according to the Beltsville Method or diluted in Kiev extender and inseminated on the day of collection. Twelve boars of the Dutch Landrace and Dutch Large White breeds were used. Farrowing rates, total number of pigs per litter and number of live pigs per litter were higher (P less than .0001) for sows inseminated with fresh semen than for sows inseminated with frozen-thawed semen (79.1%, 10.6 and 9.9 vs 47.0%, 7.4 and 7.1, respectively). Farrowing rates for sows inseminated with frozen-thawed semen were higher when semen from Dutch Large White boars was used than when semen from Dutch Landrace boars was used (58.6 vs 40.9%); the pattern reversed for insemination with fresh semen (76.5 vs 81%). Boar differences based on farrowing rate ranged from 62 to 92% for fresh semen and from 29 to 72% from frozen semen. There was no inseminator effect or farm effect on farrowing rate. On the basis of these results, frozen semen used for artificial insemination under practical circumstances can be expected to result in a farrowing rate about 30 percentage points lower and a litter size about three pigs smaller than does fresh semen.

Animals↗

Use of boar spermatozoa for artificial insemination. III. Fecundity of boar spermatozoa stored in Beltsville liquid and Kiev extenders for three days at 18 C.

A field trial was conducted on several hundred farms in The Netherlands to compare the fertilizing capacity of boar spermatozoa stored for 1, 2, or 3 d at 18 C in either Kiev or Beltsville liquid extender (BL-1). Two storage volumes (25 and 100 ml) were used. Twelve Dutch Landrace boars, all in routine commercial artificial insemination service, provided the semen in twice-a-week collections. Over an 8-wk period in March and April, 1979, 2,849 sows and gilts were inseminated. Farrowing rates were higher (P less than .0001) with Kiev extender than with BL-1 extender (69.3 vs 60.5%) when semen was used over a 3 d period. The volume at which the semen was extended and stored had no effect on any parameter. Farrowing rate declined significantly when semen stored at 18 C for more than 2 d was used (74.5% vs 64.7% with semen stored for 1 and 3 in Kiev extender; 65.9% vs 52.7% with semen stored for 1 and 3 d in BL-1 extender). The farrowing rate, total number of pigs/litter and live pigs/litter were significantly higher for sows than for gilts inseminated with similar extended semen. On the basis of the results of this trial, Kiev extender appears to be more suitable as a diluent for liquid semen than does BL-1. AI with semen extended and stored for 3 d at 18 C can be expected to result in a lower farrowing rate than AI with semen stored for 1 or 2 d.

Animals↗

Effects of pentoxifylline in the hamster zona-free oocyte spermatozoa penetration assay and on spermatozoa transmembrane migration motility.

The effects of pentoxifylline on capacitation, acrosome reaction and motility of spermatozoa in vitro were evaluated by the hamster zona-free oocyte penetration assay and the transmembrane migration method, respectively. The improvement in penetration capacity by the addition of pentoxifylline 40 min before coincubation had a trend toward statistical significance (0.05 less than less than 0.1) in normal fertile men and had statistical significance (p less than 0.05) in asthenospermic patients. There was no improvement by the addition of pentoxifylline 16 h before coincubation. Pentoxifylline significantly increased sperm motility in both normal (p less than 0.001) and asthenospermic patients (p less than 0.05) in the transmembrane migration method. This study showed that pentoxifylline could be used as a stimulant of in vitro sperm motility in male infertility.

Animals↗