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Artificial insemination homologous with oligospermic semen separated on albumin columns.

Semen samples obtained from 27 infertile men were separated on human serum albumin (HSA) columns prior to artificial homologous insemination (AIH). The columns contained either a single 7.5% HSA layer or two 17.5% and 7.5% HSA layers. Separated specimens were free of seminal debris, had significantly improved motility and progressive drive, and had a decreased percentage of abnormal forms. The total sperm count was also significantly decreased after separation, especially with the two-layer technique. No conceptions occurred in 21 couples when two-layer separation was used, but four pregnancies in 12 couples resulted from AIH with semen separated on the single-layer column. The mean total count of motile sperm obtained for AIH was 5 million with the two-layer and 31 million with the one-layer technique.

Adult↗

In vitro sperm cervical mucus penetration: studies in human and bovine cervical mucus.

The objective of this study was to assess the feasibility of substituting bovine estrus cervical mucus (BCM) for human midcycle cervical mucus (MCM) for investigation of sperm cervical mucus (CM) interaction and evaluation of cervical factor in infertility. In vitro sperm cervical mucus penetration tests (ICMPT) were performed with the use of ejaculated human spermatozoa collected from fertile donors and infertile men. Tests were performed in a flat capillary tube with the use of MCM and BCM. Mean +/- standard deviation (SD) of sperm penetration in MCM was 32.3 +/- 15.5 mm and in BCM 27.3 +/- 12.1 mm after 90 minutes of migration. The difference was significant (P less than 0.001). A highly significant correlation was found between sperm penetration in MCM and BCM. The depth of sperm penetration in both human cervical mucus (HCM) and BCM was also significantly related to sperm count and motility. Positive correlation was observed between normal morphologic features and sperm migration in BCM but not in MCM. It is concluded that human sperm penetrate BCM at a slower rate than MCM, but BCM can be effectively substituted for HCM in laboratory testing of sperm penetration.

Animals↗

Semen analyses in 1,283 men from the United States over a 25-year period: no decline in quality.

OBJECTIVE: To determine whether semen quality has changed in the United States over the last 25 years. DESIGN: Retrospective review. SETTING: Three U.S. sperm banks, Cryogenic Laboratories, Inc. (Roseville, Minnesota), Idant Laboratories (New York, New York), and California Cryobank, Inc. (Los Angeles, California). INTERVENTION: None. MAIN OUTCOME MEASURES: Age at sample collection, sperm concentration, volume, motility, and days of abstinence before sample collection were determined for each man. Linear and multiple regression analyses were used to assess changes in these characteristics over time. RESULTS: Controlling for the effects of age and duration of abstinence, there was a slight but significant increase in mean sperm concentration but no change in either motility or semen volume over the 25-year period. Both sperm motility and semen volume decreased with increasing age at sample collection. Both sperm concentration and semen volume increased as a function of duration of abstinence. There were significant differences in mean (+/- SEM) sperm concentrations (10(6) sperm/mL) and motilities between the different sperm banks with California lowest (72.7 +/- 3.1, 51.4% +/- 1.1%, respectively), Minnesota higher (100.8 +/- 2.9, 56.0%, respectively), and New York highest (131.5 +/- 3.5, 58.2% +/- 0.5%, respectively). CONCLUSIONS: Our data show no decline in sperm counts over a 25-year period in 1,283 men who banked sperm before vasectomy at three distinct geographical sites in the United States.

Adult↗

Identification of sperm subpopulations with specific motility characteristics in stallion ejaculates.

The aim of this study was to test the presence of separate sperm subpopulations, with specific motility characteristics, in stallion ejaculates by using a computer-assisted semen motility analysis (CASA) system. Motility data were analyzed with a hierarchical clustering of variables based on a correlation or covariance matrix to select like parameters of sperm motility descriptors that better explain the kinetics of spermatozoa. The statistical analyses clustered the whole motile sperm population in both fresh and 24 h stored ejaculates into four separate groups. There were significant differences in the distribution of the four subpopulations (P < 0.001) as well as in the total sperm number and the percentage of total motility (P < 0.01) in fresh semen among the five stallions tested. Our results show that separate subpopulations of spermatozoa with different motility characteristics coexist in stallion ejaculates. These subpopulations were maintained, although with a less-progressive motion pattern, after 24 h of storage. The study of subpopulations in ejaculates that have confirmed fertilizing capacity showed that the majority of the motile spermatozoa in these ejaculates are included in a subpopulation with high progressive motility and low linearity, and the ejaculates with proven fertility that have a total sperm count > or = 20 x 10(9) spermatozoa/ejaculate show all of their motile sperm included in this subpopulation. Our results show that the use of the CASA system is a relatively simple approach to the study of sperm subpopulation patterns in equine ejaculates.

Animals↗

Effect of semen collection practices on sperm characteristics before and after storage and on fertility of stallions.

This study analyzed effects of different methods and intervals of semen collection on the quantity and quality of fresh, cool-stored, and frozen-thawed sperm and fertility of AI stallions. In Experiment 1, ejaculates were obtained from six stallions (72 ejaculates per stallion) using fractionated versus non-fractionated semen collection techniques. Initial sperm quality of the first three jets of the ejaculate was not different from that of total ejaculates. Centrifugation of sperm-rich fractions before freezing improved post-thaw motility and sperm membrane integrity when compared to non-centrifuged sperm-rich fractions or non-fractionated centrifuged ejaculates (P<0.05). In Experiment 2, semen from four stallions (60-70 ejaculates per stallion) was collected either once daily or two times 1h apart every 48 h. The first ejaculates of double collections had significantly higher sperm concentrations, percentages of progressively motile sperm (PMS) after storage for 24h at 5 degrees C and lower percentages of midpiece alterations than single daily ejaculates. Semen collected once daily showed significantly lower values of live sperm after freezing and thawing than the first ejaculate of two ejaculates collected 1h apart every 48 h. In Experiment 3, semen was collected from 36 stallions (> or =12 ejaculates per stallion) during the non-breeding season and the time to ejaculation and the number of mounts was recorded. When time to ejaculation and the number of mounts increased, volume and total sperm count (TSC) also increased (P<0.05), whereas a decrease was observed in sperm concentration, percentage of PMS after storage for 24 h at 5 degrees C, percentage of membrane-intact sperm in fresh semen (P<0.05) as well as motility and percentage of membrane-intact sperm of frozen-thawed sperm (P<0.05). In Experiment 4, AI data of 71 stallions were retrospectively analyzed for the effect of number of mounts per ejaculation and frequency, time interval of semen collections on pregnancy, and foaling rates (FRs) of mares. Semen volume increased, but sperm concentration and percentage of PMS after 24-h cool-storage decreased with increasing number of mounts on the phantom (P<0.05). A statistically significant inter-relationship was demonstrated between frequency and interval of semen collection and FR. Mares inseminated with stallions from which semen was collected frequently (> or =1 on an average per day) showed significantly higher FRs than mares inseminated with semen from stallions with a daily collection frequency of 0.5-1 or <0.5. FR of mares inseminated with stallions having 0.5-1 days between semen collections was significantly better than FR of mares that were inseminated with stallions having semen collection intervals of 1-1.5 days or >2.5 days.

Animals↗

Effects of sauna on sperm movement characteristics of normal men measured by computer-assisted sperm analysis.

The effects of sauna exposure on sperm movement characteristics and other semen parameters were evaluated using computer-assisted sperm analysis (CASA). A significant (p < 0.01) decrease in average path velocity (VAP), curvilinear velocity (VCL) and amplitude of lateral head displacement (ALH) was found after exposure to sauna for 2 weeks. The altered parameters returned to their original values within 1 week after cessation of sauna exposure. Mean values for semen volume, sperm count, percentage motility, sperm morphology and sperm penetration assay (SPA) were not statistically different during and after sauna, when compared to the corresponding control values. The results suggest that increasing scrotal temperature by sauna causes a reversible decrease in sperm movement parameters.

Adult↗

Assay of testicular angiotensin-converting enzyme activity in human spermatozoa.

The testicular isozyme of angiotensin-converting enzyme (ACE) is associated with male fertility. Spermatozoa from mice lacking ACE showed defects in transport within the oviducts and in binding to zonae pellucidae although the animals had normal sperm count, morphology and motility. In fact, unexplained infertility is difficult to be predicted by conventional parameters such as sperm count. We measured membrane testicular ACE activity in a sperm suspension in PBS and total testis ACE activity in spermatozoa by solubilization with Triton X-100. Total testis ACE activity and membrane testis ACE activity of the same subject were compared in 12 control subjects. We demonstrated that testicular ACE is stable in spermatozoa and the assay of testicular ACE activity is possible. Total testicular ACE activity was approximately twice the membrane testicular ACE activity in all of the subjects tested. The assay of testicular ACE activity in human spermatozoa could be a new method for the assessment of sperm function.

Adult↗

Epididymal and sex accessory gland secretions in transfusion-dependent beta-thalassemic patients: evidence of an impaired prostatic function.

Neutral alpha-glucosidase levels as epididymal marker, fructose levels as vesicular marker, zinc, citric acid and prostate specific antigen levels as prostatic markers were measured in the seminal plasma of eight transfusion-dependent beta-thalassemic patients in order to study epididymal and sex accessory gland secretions (eighteen subjects served as controls). FSH and LH as well as total and free testosterone were detected displaying unaltered serum values. Ejaculate of patients showed normal sperm count and low sperm motility, in the meantime seminal plasma exhibited unaltered both neutral alpha-glucosidase and fructose values but low levels of zinc, citric acid and prostate specific antigen were noticed as well. These data suggest an impaired prostatic secretion in the thalassemic patients studied. A local iron toxicity on the prostatic tissue could be supported by the decrease of its specific markers observed only in the subgroup of patients with high ferritin serum levels.

Adolescent↗

Chronic cadmium toxicity to sperm of heavy cigarette smokers: immunomodulation by zinc.

The aim of the study was to investigate the role of zinc therapy in 125 male cigarette smokers with infertility. The mechanism involved in the zinc/cadmium relationship was evaluated through the effect of a zinc-deficient diet and supplementation on testes of male adult Sprague-Drew rats. Heavy smoking was associated with low sperm count, motility, and morphology and increased seminal cadmium levels. Zinc therapy improved sperm quality and increased seminal IL-4, but reduced TNF-alpha and IFN-gamma. A zinc-deficient diet led to high cadmium testicular accumulation comparable with those supplemented with cadmium. Cadmium had a linear correlation with TNF-alpha and IFN-gamma, but not with IL-4. Cytology of testicular aspirate and histopathology were normal in supplemented groups as in controls. These results indicate that zinc modulates the putative effect of cadmium through its enhancement of T-helper 2 cytokines expression and down-regulation of T-helper 1 cytokines.

Animals↗

Gene deletions in an infertile man with sperm fibrous sheath dysplasia.

BACKGROUND: Asthenozoospermia may sometimes be related to genetic structural defects of the sperm tail detectable by transmission electron microscopy. Dysplasia of the fibrous sheath (DFS) is a genetic sperm defect, characterized by dysplastic development of the axonemal and periaxonemal cytoskeleton. We report the case of an infertile man with normal sperm count and total sperm immotility in which dysplasia of the fibrous sheath, Akap3, Akap4 gene deletions, meiotic segregation of chromosomes 18, X and Y and Y microdeletions were investigated. METHODS: A 32-year-old man with a 3-year history of primary infertility presented at our Regional Referral Center for Male Infertility. Family medical history, lymphocyte karyotype, PCR analysis, physical examination, hormone assays and semen analysis were performed. RESULTS: Ultrastructural sperm evaluation showed dysplasia of the fibrous sheath. Immunostaining of AKAP4 protein was negative in sperm tails. PCR analysis revealed intragenic deletions of the Akap3 and Akap4 genes. Fluorescence in situ hybridization on sperm showed a high frequency of XY disomy. CONCLUSION: In this infertile patient, our results suggest a possible relationship between dysplasia of the fibrous sheath, partial deletions in the Akap3 and Akap4 genes and absence of AKAP4 protein in the fibrous sheath. These findings, however, were not detected in another four patients with dysplasia of the fibrous sheath. Our results require future confirmatory molecular analyses.

A Kinase Anchor Proteins↗

Chromosomal segregation in spermatozoa of 14 Robertsonian translocation carriers.

Male carriers of Robertsonian (Rob) translocations can have fertility problems associated with low sperm counts and abnormal sperm morphology. In this study, spermatozoa from 14 Rob translocation carriers, seven der(13;14), two der(13;15), two der(14;15), two der(14;21) and one der(21;22), were tested by fluorescence in-situ hybridization (FISH) for the chromosomes involved, to study meiotic segregation behaviour. It was shown that in each type of Rob translocation, meiotic segregation behaviour is similar, comparable and occurs non-randomly. Most of the spermatozoa results from alternate segregation (range: 76-89.47%). There is, however, still much unbalanced spermatozoa resulting from adjacent segregation mode (range: 10.24-23.41%). These data provide useful information for genetic counselling purposes. Moreover, aneuploidy for chromosomes 13,18, 21, X and Y was studied in five patients and suggested an inter-chromosomal effect.

Chromosome Segregation↗

Sperm-macrophage interaction in the mouse: a quantitative assay in vitro using 111indium oxine-labeled sperm.

The role of reproductive tract macrophages in contraception and reproductive failure has become widely recognized. However, in vitro analysis of sperm phagocytosis by macrophages has relied upon a semi-quantitative method of sperm counting that is of limited accuracy and reproducibility. We have developed an assay using murine sperm labeled with 111indium oxine, and results indicate the labeling to be rapid and efficient. Incorporation of 111indium into sperm increased the dose and sperm concentration and reached 90% maximal uptake after 15 min incubation, with maximal uptake occurring at 30 min. No decrease in sperm motility was noted with levels of oxine in excess of those required for significant labeling. Maximal labeling efficiency occurred in phosphate-buffered saline (PBS), with Dulbecco's modified Eagle's medium (DMEM) + 10% adult bovine serum (ABS) producing significantly less uptake. Label dissociation was detectable in PBS at room temperature, but at 37 degrees C in DMEM + 10% ABS, loss of label occurred at a rate of 23.5%/h. Addition of labeled sperm to murine macrophage monolayers under optimal conditions resulted in uptake of 111indium by macrophages, while free label was unincorporated. Results indicated assay specificity for macrophage-limited uptake, with insignificant label uptake by nonphagocytic murine fibroblasts and better sensitivity than sperm counting. Macrophages from Bacillus Calmette-Guerin (BCG)-infected mice resulted in a decrease in sperm uptake. Female macrophages showed greater capacity for sperm uptake than those of the male mouse. These initial studies demonstrated the utility of this model system in enhancing the understanding of sperm-macrophage interaction in the female reproductive tract.

Animals↗

Antispermatogenic activity of Morinda lucida extract in male rats.

AIM: To investigate the effect of Morinda lucida Benth (Rubiaceae) on the reproductive activity of male albino rats. METHODS: Two groups of rats were treated with 400 mg/(kg .d) of Morinda lucida leaf extract for 4 and 13 weeks, respectively. The control rats received the vehicle. All the treated rats had corresponding recovery groups. At the end of each experimental period, animals were killed and organ weights, sperm characteristics, serum testosterone levels, histology of the testes and fertility were assessed. RESULTS: Morinda lucida leaf extract did not cause any changes in body and somatic organ weights, but significantly increased the testis weight (P 0.05). The sperm motility and viability, and the epididymal sperm counts of rats treated for 13 weeks were significantly reduced (P 0.05). Sperm morphological abnormalities and serum testosterone levels were significantly increased (P 0.05). There were various degrees of damage to the seminiferous tubules. The extract reduced the fertility of the treated rats by reducing the litter size. Reversal of these changes, however, occurred after a period of time. CONCLUSION: The extract of Morinda lucida has reversible antispermatogenic properties.

Animals↗

Reproductive function in male endurance athletes: sperm analysis and hormonal profile.

The purpose of this investigation was to study the effects of endurance exercise on male reproductive function (sex hormones and seminograms). Professional cyclists [n = 12; mean age 24 +/- 2 (SD) yr], elite triathletes (n = 9; 26 +/- 3 yr), recreational marathon runners (n = 10; 32 +/- 6 yr), and sedentary subjects (control group; n = 9; 30 +/- 4 yr) were selected as subjects. for each group, the following parameters were measured three times during the sports season (training period: winter; competition period: spring; resting period: fall): percentage of body fat, hormonal profile (resting levels of follicle-stimulating hormone, luteinizing hormone, total and free testosterone, and cortisol), and seminograms (quantitative parameters sperm volume and sperm count; qualitative parameters: sperm motality and morphology). The following comparisons were made in the measured parameters: 1) within groups (longitudinal design) and 2) between groups in each of the three periods (cross-sectional design) and over time (mixed design). In addition, both the volume and the intensity of training of each subject during the season (except for the control group) were quantified. Despite significant differences in training characteristics and in body fat percent, in general no significant differences (P > 0.05) were found in hormonal profiles or in semen characteristics between or within groups. A lower sperm motility (46.2 +/- 19.5%), however, was observed in the cyclists during the competition period when compared either with the other groups during this same period (P < 0.05) or with themselves during the other two periods of study (P < 0.01). In any case, the later phenomenon was attributed to physical factors associated with cycling, such as mechanical trauma to the testis and/or increased gonadal temperature. In conclusion, our findings suggest that endurance exercise does not adversely affect the hypothalamic-pituitary-testis axis.

Adult↗

Methods for evaluating the effects of environmental chemicals on human sperm production.

Sperm tests provide a direct and effective way of identifying chemical agents that induce spermatogenic damage in man. Four human sperm tests are available: sperm count, motility, morphology (seminal cytology) and the Y-body test. These sperm tests have numerous advantages over other approaches for assessing spermatogenic damage, and they have already been used to assess the effects of at least 85 different occupational, environmental, and drug-related chemical exposures. When carefully controlled, seminal cytology appears to be statistically more sensitive than the other human sperm tests and should be considered an integral part of semen analysis when assessing induced spermatogenic damage. Human sperm studies have complex requirements and, before sampling, careful consideration should be given to exposure details, group size and makeup, as well as animal and human data that indicate spermatogenic effects. Several study designs are possible and should include questionnaires covering medical and reproductive histories as well as known confounding factors. Animal sperm tests, such as the mouse morphology test, may be used to identify the toxic components of a complex mixture. Animal tests may also help assess the chemical effects on fertility and reproductive outcome in cases when human data are incomplete. Further efforts are needed in these areas to develop improved human sperm tests sensitive to induced spermatogenic damage, to develop improved animal models of induced spermatogenic damage, to understand the relationships among sperm changes, fertility, and reproductive outcome, and to develop sperm tests with express mutational end points.

Animals↗

Environmental toxicology and health effects associated with dinitrotoluene exposure.

Dinitrotoluenes (DNTs) are nitroaromatic compounds appearing as pale yellow crystalline solids at room temperature. Dinitrotoluenes exist as a mixture of 2 to 6 isomers, with 2,4-DNT, and 2,6-DNT being the most significant. About 500 persons are estimated to be potentially exposed yearly to 2,4-DNT and 2,6-DNT during the production of munitions and explosives. The main route of human exposure at ammunition facilities is inhalation, but dermal contact and inadvertent ingestion can also be substantial. In factory workers, exposure to DNTs has been linked to many adverse health effects, including cyanosis, vertigo, headache, metallic taste, dyspnea, weakness and lassitude, loss of appetite, nausea, and vomiting. Other symptoms including pain or parasthesia in extremities, abdominal discomfort, tremors, paralysis, chest pain, and unconsciousness have also been reported. The primary targets of DNT toxicity are the hematopoietic system (pallor, cyanosis, anemia, and leukocytosis), the cardiovascular system (ischemic heart disease), the nervous system (muscular weakness, headache, dizziness, nausea, insomnia, and tingling pains in the extremities) and the reproductive system (reduction of sperm counts, alteration of sperm morphology, and aspermatogenesis). An association between DNT exposure and increased risk of hepatocellular carcinomas and subcutaneous tumors in rats, as well as renal tumors in mice, has been established. Epidemiologic studies of DNT toxicity have been limited to small groups of workers who had been occupationally exposed at various ammunitions production facilities. Clearly defining the health effects of DNTs with a high degree of confidence has therefore been difficult because of the multigenic nature of occupational exposure. In an attempt to update the toxicologic profile of the DNTs, we hereby provide a critical review of the environmental and toxicologic pathology of DNTs, with a special emphasis on their potential implications for public health.

Air Pollutants, Occupational↗

Sperm transport in the human female reproductive tract in relation to semen analysis characteristics and time of ovulation.

Laparoscopic sperm recovery from the pouch of Douglas and tubal fimbriae was performed in 64 infertile couples. Spermatozoa were recovered from 16/35 couples investigated after AIH, and from 13/29 couples post coitum. The method of insemination had no effect on the result, which was positive in 45.3% of all couples, although AIH did result in significantly larger numbers of peritoneal spermatozoa. The number of peritoneal spermatozoa did not show any direct correlation with the number inseminated, but there were reductions along the tract of 5.83 (+/- 1.4 s.d.) orders of magnitude for total sperm count, and 5.52 (+/- 1.21 s.d.) for the number of motile spermatozoa. Only sperm motility had a significant influence on the success of sperm transport; spermatozoa were recovered from patients with sperm densities as low as 3.0 and 3.5 x 10(6)/ml, but with 56 and 44% motile spermatozoa. No influence of cycle day within the range +/- 4 days of ovulation on sperm transport was found. In 45 couples, routine semen analyses were apparently completely normal, but the incidence of sperm recovery was still only 49% (22/45), suggesting that a failure of sperm transport may have been a significant causative factor in their infertility.

Female↗