Cytogenetics of somatic cells and sperm from a 46,XY/45,X mosaic male with moderate oligoasthenoteratozoospermia.

OBJECTIVE: To determine aneuploidy frequencies in sperm from a patient with normal phenotype and 46,XY/45,X mosaicism in somatic cells (peripheral lymphocytes). DESIGN: Case report. SETTING: Infertility clinic and genetics laboratory. PATIENT: A 30-year-old male with primary infertility and moderate oligoasthenoteratozoospermia. INTERVENTION(S): Cytogenetic analysis of somatic cells and determination by fluorescence in situ hybridization of aneuploidy frequencies for the gonosomes (sex chromosomes) and chromosome 18 in sperm from whole and Percoll-separated semen. MAIN OUTCOME MEASURE(S): Somatic and gametic aneuploidy were scored. RESULT(S): Analysis of lymphocyte metaphase cells showed a mosaic 46,XY (90%)/ 45,X (10%) karyotype. Significantly higher frequencies of gonosomal (semen, 1.92% versus 0.70%; Percoll, 1.12% versus 0.46%), and chromosome 18 (semen, 0.89% versus 0.28%; Percoll, 0.26% versus 0.10%) disomy were detected in the sperm of the patient compared with those observed in spermatozoa from a proved fertile control. CONCLUSION(S): Significantly higher frequencies of aneuploid sperm suggest that the patient is at elevated risk of producing offspring with numerical chromosome abnormalities.

Comparison between swim-up and glass bead column techniques for the separation of human spermatozoa.

The glass bead column filtration method was compared with the swim-up technique regarding spermatozoa recovery in normal and pathological human semen. The general characteristics of the semen and the spermatic ATP concentration were measured in 67 patients. Samples processed with glass bead columns provided a good amount of motile forms which however, showed less velocity and linearity than in the case of using the swim-up technique. Additionally, the ATP determination in spermatozoa was included.

Computer assisted measurement in normal and pathological human semen, fresh and post swim-up technique.

A total of 429 semen samples were studied. Two hundred and twenty-nine samples with normal characteristics were processed following a basic procedure; the remaining 200 were normal and pathological samples that were analyzed pre and post swim-up. Semen specimens were allowed to liquefy for 30 minutes and sperm count, motility, velocity and linearity were determined using the Cellsoft Automatic Semen Analyzer. In normal patients, a significant increase of motility, velocity and linearity (p less than 0.001) post swim-up, was observed. Sperm recovery in this group was 13.9 +/- 1% of the whole motile sperm population. In the polyzoospermic group, recovery of motile spermatozoa post swim-up was significantly decreased as compared with the normal group (p less than 0.001). In the asthenozoospermic group (plus either hypospermia or hyperspermia, or with more than 30% of motile spermatozoa or less than 30% of motility), no variations of velocity or linearity as compared to the normal group were observed. In all the pathological groups studied, a significant increase in velocity and linearity (p less than 0.001) post swim-up, was observed.

Effect of lipid peroxidation upon human spermatic adenosinetriphosphate (ATP). Relationship with motility, velocity and linearity of the spermatozoa.

The aim of the present paper was to study the effect of forced lipid peroxidation upon human spermatozoa and its relation with motility, velocity, linearity and ATP content. Forty-three pools of semen samples were studied one hour after liquefaction. After washing, normal and pathological spermatozoa presented no variations in motility, velocity or linearity with respect to basal values. Forced peroxidation produced a significant decrease of motility, velocity and linearity (p less than 0.02) with respect to control. Spermatic ATP was not significantly different in control vs forced peroxidation groups in normozoospermic samples. In contrast, a significant decrease of ATP between control and peroxidation groups was observed in asthenozoospermia at 120 minutes of incubation (p less than 0.02). In the asthenozoospermic group, a significant LPO-p increase was observed in comparison with the normal group.

Dysplasia of the fibrous sheath: an ultrastructural defect of human spermatozoa associated with sperm immotility and primary sterility.

A study of a group of five patients presenting with primary sterility and showing severe sperm immotility is presented. Most spermatozoa in these patients showed rigid, short, thick, and/or irregular tails and 95 to 100% were immotile. Electron-microscopy disclosed a common pattern of flagellar abnormalities. There was a dysplastic development of the fibrous sheath, which appeared hyperplastic and disorganized. The axoneme was either missing or grossly distorted. In a few instances, a normal flagellum could be identified. Similar alterations also were detected in maturing spermatids, suggesting that the described defect develops during spermiogenesis. Two of the five patients had recurrent bronchial and sinusal infections and bronchiectasis, suggesting the possible existence of an associated abnormality in respiratory cilia. The existence of a common ultrastructural defect affecting most spermatozoa, its presence in two brothers, and the possibility of association with immotile respiratory cilia point to the existence of a syndrome (namely the "dysplasia of the fibrous sheath") of possible familial transmission.

New method of evaluating sperm morphology with predictive value for human in vitro fertilization.

A prospective study was planned to evaluate sperm morphology as a parameter to predict the fertilization outcome in an in vitro fertilization program. Couples applying to in vitro fertilization were admitted into this project when the sperm concentration was greater than 20 million per mL and motility greater than 30 per cent. Based on new strict criteria for evaluating normal sperm morphology, patients were divided prospectively into 2 groups. In group I (25 patients) normal sperm morphology was less than 14 per cent, and in group II (71 patients) normal sperm morphology was greater than 14 per cent, using a threshold established previously. Multiple regression analysis was used to evaluate different parameters: concentration, motility, and morphology against the dependent variables, fertilization, and cleavage. The only factor which was significantly correlated with fertilization and cleavage was normal sperm morphology (P less than 0.0001). The fertilization rate (per oocyte) and the cleavage rate were 49.4 per cent and 47.6 per cent in group I and 88.3 per cent and 87 per cent in group II (P less than 0.0001). The ongoing pregnancy rate per laparoscopy and per embryo transfer was 4 per cent and 5.5 per cent, respectively, in group I and 18.3 per cent and 18.5 per cent, respectively, in group II (no significant difference). This study demonstrates the value of analyzing sperm morphology using the criteria recommended in terms of predicting fertilization and perhaps pregnancy outcome. Patients can be better counseled and the probability of fertilization or no fertilization can be more accurately established. Furthermore a trend is shown in the pregnancy rate that may indicate the importance of the male genome in establishing a pregnancy.

Adenosine 5 triphosphate (ATP) content and acrosin activity in polyzoospermic subjects.

The aim of this paper was to measure the concentration of spermatic ATP and acrosin in a group of patients with polyzoospermia, and to compare these values with those found in a group of normozoospermic men. The general characteristics of the semen, the ATP and the acrosin, were measured in a group of 46 men with polyzoospermia, who consulted in our clinic for sterility. The results were compared with those of 50 normozoospermic men in the control group. Statistically, no significant difference was found between the volume, motility and morphology of the samples in both groups; however, the concentration of spermatic ATP and acrosin decreased significantly in a group of polyzoospermic patients. This could be due to a functional defect of the spermatozoa in this group of patients.

Lack of a head in human spermatozoa from sterile patients: a syndrome associated with impaired fertilization.

Three patients with primary sterility in whom the majority of spermatozoa lacked a normally implanted head are presented. A small cephalic knob was evident in most of them by routine colorimetric techniques, and the Feulgen reaction failed to show any deoxyribose nucleic acid. The morphologic features of the tails was normal. Few loose sperm heads were observed in the ejaculates. Even though motility was decreased, there were numerous acephalic sperms with different degrees of forward motility. Electron microscopy showed a well-organized structure of the centrioles and connecting piece, which were located in the neck region within a small cytoplasmic mass, but no chromatin was detected in any case. Studies on immature spermatids present in semen evidenced an independent anomalous development of heads and tails and suggested that they became separated at the end of spermatid maturation. This anomaly, of probable genetic origin, is interpreted to be due either to an alteration in the mechanism of migration and positioning of the tail on the caudal pole of the nucleus or to an interference with the formation of the implantation fossa of the head, which normally accommodates the connecting piece.

A quick, reliable staining technique for human sperm morphology.

The evaluation of sperm morphology is still an important parameter in the diagnosis of the infertile male. Most techniques used for staining human sperm are very time-consuming. A routine stain used for determining differential count of leucocytes (Diff-Quik stain) was evaluated against the standard Papanicolaou stain. Morphology results from 20 duplicate semen smears using both staining methods were determined separately by 2 technicians using a blind protocol. No significant differences were observed when comparing the two staining methods (paired Student's t-test). The advantages of the Diff-Quik stain are: a) complete staining-to-reading time under 7 min, b) commercially prepared reagents, and c) case of staining procedure.

Adenosine 5'-triphosphate (ATP) concentration and acrosin activity in human spermatozoa. Their relation with the sperm penetration assay (SPA).

Fertility of men depends on the quality of semen. The aim of the present paper is to determine both the acrosin activity by radioimmunoassay and ATP concentration by bioluminescence in human spermatozoa, and evaluate these results in those samples with normal or low sperm penetration according to SPA test. Ejaculates obtained from 42 untreated men, were studied one hour after the obtention. These materials were divided into two groups:20 human semen samples with "in vitro" potentiality to penetrate zone-free hamster ova, between 15% to 98% and 22 human semen with SPA test between 0% to 14%. When we compare the group with normal penetration response vs that group with low or absent penetration ones, a significant decrease of ATP and acrosin concentrations was observed (P less than 0.001). Nevertheless no significant difference was observed in relation with percentage of motility, volume (ml), sperm concentration (10(6)/ml), percent of quick progressive spermatozoa and number of gametes capable of migrating into the medium layer (10(6)), between the group with low or absent penetration test against that one with normal zona-free hamster egg test.

Adenosine 5'-triphosphate (ATP) activity in normal and pathological human semen. Relationship with round cells of the ejaculate.

The aim of the present paper is deleted to determine ATP activity in the human seminal cells by bioluminescence and to investigate the relationship between these values and the number of round cells either peroxidase positive or peroxidase negative. We studied 244 untreated men, divided into six groups. Our mean value of ATP activity in normal semen was 20.02 +/- 0.65 n moles per 10(8) spermatozoa. In asthenozoospermic or oligoasthenozoospermic patients with less than 1,000,000 round cells per ml, the concentration of ATP was significantly lower than normal (P less than 0.001) or (P less than 0.05). Semen with normal characteristics or samples from asthenozoospermic or oligoasthenozoospermic patients but with more than 1,000,000 round cells, per ml and with predominance of peroxidase negative cells, had an ATP concentration higher than normal subjects (P less than 0.001). We believe that the knowledge of the quantity and quality of round cells of the ejaculate is important to interpret the seminal values of ATP.

Cytogenetic findings in 301 sterile and/or infertile men with abnormal spermograms

En los últimos años se ha establecido que las anomalias cromosómicas constituyen una de las causas de esterilidad y/o infertilidad, por producir bloqueo de la meiosis (dando lugar a oligozoospermia severa o azoospermia) o formar gametas genéticamente anormales no viables o fecundantes, las cuales originan cigotas anormales que son eliminadas antes o después de la implantación o bien dar lugar a nacimientos de niños malformados. En pacientes varones que consultan por esterilidad y/o infertilidad la frecuencia de anomalías cromosómicas mayores varía desde un 2,2 hasta un 15%, con una frecuencia promedio del 8,6%, cifra claramente mayor que la encontrada en la población general que es de 0,6%. Además, existe una estrecha correlación entre frecuencia de anomalías cromosómicas y grado de oligozoospermia. Sin embargo, no siempre es posible conocer el mecanismo por el cual una anomalía cromosómica altera la espermatogénesis. La expresión de estas anomalías, fundamentalmente los reordenamientos equilibrados, son variables y no predecibles, variando desde fertilidad normal hasta esterilidad total con severa oligozoospermia o azoospermia. El motivo del presente trabajo es proveer datos adicionales acerca de la incidencia y tipos de anomalías mayores y menores en varones estériles y/o infértiles constituyendo el estudio cromosómico la primera evaluación diagnóstica luego de dos espermogramas anormales. Los estudios cromosómicos se realizaron con metodología convencional y técnicas de bandeo G y C. Se estudiaron un total de 301 pacientes. Setenta y nueve por ciento consultaron por esterilidad primaria 7% esterilidad secundaria y 14% por infertilidad. Veintitrés por ciento eran azoospérmicos, 13% oligozoospérmicos severos, 56% oligozoospérmicos y oligoastenoteratozoospérmicos y 8% astenozoospérmicos y/o teratozoospérmicos...

Cytogenetic findings in 301 sterile and/or infertile men with abnormal spermograms

En los últimos años se ha establecido que las anomalias cromosómicas constituyen una de las causas de esterilidad y/o infertilidad, por producir bloqueo de la meiosis (dando lugar a oligozoospermia severa o azoospermia) o formar gametas genéticamente anormales no viables o fecundantes, las cuales originan cigotas anormales que son eliminadas antes o después de la implantación o bien dar lugar a nacimientos de niños malformados. En pacientes varones que consultan por esterilidad y/o infertilidad la frecuencia de anomalías cromosómicas mayores varía desde un 2,2 hasta un 15%, con una frecuencia promedio del 8,6%, cifra claramente mayor que la encontrada en la población general que es de 0,6%. Además, existe una estrecha correlación entre frecuencia de anomalías cromosómicas y grado de oligozoospermia. Sin embargo, no siempre es posible conocer el mecanismo por el cual una anomalía cromosómica altera la espermatogénesis. La expresión de estas anomalías, fundamentalmente los reordenamientos equilibrados, son variables y no predecibles, variando desde fertilidad normal hasta esterilidad total con severa oligozoospermia o azoospermia. El motivo del presente trabajo es proveer datos adicionales acerca de la incidencia y tipos de anomalías mayores y menores en varones estériles y/o infértiles constituyendo el estudio cromosómico la primera evaluación diagnóstica luego de dos espermogramas anormales. Los estudios cromosómicos se realizaron con metodología convencional y técnicas de bandeo G y C. Se estudiaron un total de 301 pacientes. Setenta y nueve por ciento consultaron por esterilidad primaria 7% esterilidad secundaria y 14% por infertilidad. Veintitrés por ciento eran azoospérmicos, 13% oligozoospérmicos severos, 56% oligozoospérmicos y oligoastenoteratozoospérmicos y 8% astenozoospérmicos y/o teratozoospérmicos...(AU)

Effects of methyl xanthines upon the spermatozoa adenosinetriphosphate (ATP) concentration in normal semen.

The aim of the present study was to ascertain the effect of methyl xanthines upon the spermatozoa ATP concentration in normal semen in vitro. 26 normal semen were studied. The specimens were diluted after liquefaction with equal volume of Lopata's buffer or Lopata's buffer plus 1 or 6 mM either of caffeine or pentoxifylline. The samples of semen were incubated at room temperature during 90, 180 and 240 minutes before motility and ATP determination by the firefly luciferin-luciferase method. Significant variation was observed in sperm motility nevertheless variations in the ATP concentration was not induced by any of the methyl xanthines we used.