Biomedical subjects

J Benet

Publications and source records attributed to J Benet.

At least 37 records · Page 2Linked to original sources

PubMed2000

Correlation between centromere and chromosome length in human male pronuclear chromosomes: ultrastructural analysis.

Ultrastructural and morphometric analyses of centromeric regions by scanning and transmission electron microscopy have been performed in chromosomes from male pronuclei obtained by heterologous fertilisation of hamster oocytes with human spermatozoa. In 1308 of 1323 chromosomes analysed, the primary constriction showed a defined biconcave constriction of variable length (0.56-1.34 microns) and constant width (0.64-0.7 micron). A positive correlation was observed between centromeric length and chromosome length. In some chromosomes, the primary constriction appears as decondensed regions of variable length (1.6-2.51 microns) composed of chromatin fibres with a minimum diameter of 30 nm.

Animals↗

PubMed1999

[Haemophilus influenzae type b invasive disease in Catalonia (1996)].

BACKGROUND: The purpose of this study was to find out the incidence and characteristics of H. influenzae type b invasive disease (HibID) in Catalonia, Spain. MATERIAL AND METHODS: An active surveillance of H. influenzae isolated from normally sterile sites was carried out during 1996. Microbiology laboratories of hospitals of Catalonia were periodically contacted by telephone. The serotype of all the strains was studied. RESULTS: The incidence of H. influenzae invasive disease (HIID) was 7.1 per 100,000 in children under 5 years and 1.0 per 100,000 in those over 5 years. The incidence of serotype b was 6.4 per 100,000 children under 5 years and 0.2 above this age. Only three strains belonged to types other than b (d, e and f). CONCLUSIONS: The incidence of HIbID is uncommon in Catalonia, lower than that registered in the prevaccine era in other countries and regions of the same geographical area.

Adolescent↗

PubMed1998

Chromosome studies in first polar bodies from hamster and human oocytes.

Most studies on preconception diagnosis published so far have used polymerase chain reaction (PCR) analysis to identify single gene defects. Although fluorescent DNA probes have been used to obtain a partial cytogenetic diagnosis of aneuploidies in first polar bodies without defined chromosome structures, the analysis of structural chromosome anomalies in the interphase nucleus is not adequate. We describe a procedure to obtain first polar body chromosome complements from hamster and human oocytes. In 63.6% (105 of 165) of hamster first polar bodies the chromosome complement showed a defined chromosome morphology and in 94.1% (16 of 17) of human oocytes fixed after follicular puncture it was possible to obtain high quality, well spread chromosome complements. First polar body chromosomes are fuzzy and shorter than oocyte chromosomes, but fluorescent in-situ hybridization results obtained in human first polar bodies clearly show that it is possible to detect whole chromosomes, centromeres and unique sequences, including the terminal regions of small chromosomes. This suggests that in fresh oocytes, DNA loss resulting from apoptotic chromosome fragmentation has not yet occurred. Using the procedure described, first polar bodies could be used to analyse the meiotic segregation of maternal structural abnormalities and to detect numerical chromosome anomalies in humans.

Aneuploidy↗

PubMed1998

Sperm segregation analysis of a complex chromosome rearrangement, 2;22;11, by whole chromosome painting.

Using the human sperm-hamster oocyte fusion technique and whole chromosome painting, we studied sperm chromosome segregation in a male heterozygous for a complex chromosome rearrangement, 46,XY,-2, +der(2)t(2;11)(q13; q23),-11,+der(11)t(11;22)(q23;q11.2),-22, +der(22)t(2;22)(q13; q11.2). A total of 208 sperm complements were analyzed. The frequency of sperm carrying a normal or a balanced complement was 13.5% (9.62% and 3.85%, respectively). The frequency of unbalanced sperm was 86.5% (64.9% from 3:3 segregation, including 30 different types; 20.7% from 4:2 segregation, including 21 different types; and 0.96% from 5:1 segregation, including 2 different types). The sex ratio, determined in 134 sperm complements, did not differ from the expected 1:1 ratio. The results obtained in this study are compatible with the formation, during the synaptic process, of a complex hexavalent figure involving chromosomes 2, 11, and 22. The behavior and segregation of this complex figure would explain the high frequency (86.5%) of unbalanced complements observed in this carrier.

Animals↗

PubMed1997

Hamster origin of metaphases with multiple chromosome rearrangements in first cleavage human-hamster embryos.

Laboratories using the human sperm-hamster egg fertilization system to analyse sperm chromosomes obtain, sporadically, metaphases with multiple aberrations. Due to the high number of aberrations, these metaphases cannot be fully karyotyped. In some of them, one or several human chromosomes can be identified, guaranteeing the human origin of the whole metaphase. However, in others, none of the chromosomes can be recognized as human. This latter type of grossly rearranged metaphases is characterized by complex chromatid exchanges, multifragmented chromosomes and pulverized chromosome material. Using fluorescent in-situ hybridization techniques (FISH) with either human or hamster genomic DNA probes, we examined the origin of this second type of metaphase with multiple chromatid exchanges and fragmented chromosomes. Our study demonstrates that all of them hybridize with hamster genomic DNA probes and not with human DNA, proving their hamster origin. Since some of these metaphases seem to be diploid, we suggest that they may arise from hamster eggs that have failed to complete meiosis and have not extruded the second polar body.

Animals↗

PubMed1997

Hypomethylation of human sperm pronuclear chromosomes.

Using the methylase SssI enzyme, we have analyzed the degree of in situ methylation of human sperm pronuclear chromosomes obtained by fertilizing hamster oocytes with human sperm. Untreated (control) sperm chromosome complements showed a higher degree of in situ methylation, compared to sperm complements previously treated with 5-azadeoxycytidine or lymphocyte chromosomes. This indicates that human sperm pronuclear chromosomes have a lower degree of genomic methylation compared to that of other somatic cells. The similarity in the degree of in situ methylation of the euchromatic and heterochromatic regions of chromosomes 1, 9, 15, and 16 and the Y chromosome in human sperm does not support the existence of a possible correlation between hypomethylation and heterochromatin decondensation.

Animals↗

PubMed1996

Age at first full-term pregnancy, lactation and parity and risk of breast cancer: a case-control study in Spain.

A case-control study was carried out in Spain to assess associations between parity, lactation and age at first full-term pregnancy and breast cancer. From November 1989 to February 1992, 184 incident breast cancer histologically confirmed cases were interviewed and matched by age and residence to 184 hospitalized patients and 184 community controls selected by random digit dialing. Multiple logistic regression was used to assess the independent influence of each factor on the risk of breast cancer in relation to other factors included in the model. Age at first full-term pregnancy was associated with breast cancer risk with an estimated odds ratio of 3.5 (95% CI 1.41-9.83) for women with their first birth after 30 years in comparison with those whose first birth was before age 21. Breast cancer risk decreased with increasing number of full-term pregnancies, OR 0.3 (95% CI 0.16-0.78) for women who had had more than 3 full-term pregnancies in comparison with nulliparous women. Among parous women, the estimated OR for women with more than 3 children was 0.4 (95% CI 0.13-0.81) after allowance for age at first childbirth and lactation. The estimated OR was 2.6 (95% CI 1.4-4.7) for women with a positive history of breast cancer in first-degree relatives. Breast cancer was not associated with total duration of lactation. The study indicates that parity is an independent risk factor associated to breast cancer and that the women with a late age at first full-term pregnancy constitute a high-risk group.

Breast Neoplasms↗

PubMed1996

Chromosome aberrations in human spermatozoa treated with Ca2+ ionophore A23187.

Incorporation of A23187 ionophore into the human-hamster fertilization system clearly improves the ability of human spermatozoa to penetrate zona-free hamster oocytes. Thus, an increasing number of laboratories working in human sperm cytogenetics have substituted classical incubation with Biggers-Whitten-Whittingham (BWW) medium plus human serum albumin (HSA) by pretreatment of spermatozoa with calcium ionophore A23187 which directly induces the acrosome reaction in spermatozoa. However, there have been no formal studies on the effects of this ionophore pretreatment. To determine whether calcium ionophore could affect the cytogenetic characteristics of human spermatozoa we compared A23187-treated spermatozoa with controls (only incubated with BWW + HSA) by analysing a total of 447 sperm chromosome complements from two normal donors. Our results show that there are no statistical differences in the frequency and the types of human sperm chromosomal abnormalities between the two methods of sperm treatment. Thus, ionophore A23187 seems not to affect the cytogenetic characteristics of human spermatozoa, and the results of laboratories using either sperm capacitation in BWW + HSA or acrosome reaction by calcium ionophore can be compared.

Acrosome↗

PubMed1996

An analysis of human sperm chromosome aneuploidy.

A sperm chromosome analysis of 24 men with normal or balanced karyotypes was carried out to study the frequency of sperm chromosome aneuploidy. A total of 3,446 human sperm complements (36-315 per donor) was analyzed after in vitro penetration of hamster eggs. Two sets of donors were studied at two different centers in the United States (center 1) and Spain (center 2). The frequencies of hyperhaploidy and hypohaploidy in control donors were similar between center 1 (1.9% vs. 7.7%) and center 2 (1.8% vs. 10.3%). In carrier donors there were no significant differences between the two centers in the frequency of hyperhaploidy (0.8% vs. 1.9%), but that of hypohaploidy was significantly higher in center 2 (11.0%) than in center 1 (4.6%). A significant excess of hypohaploid complements, as compared to hyperhaploid complements, was found in both centers in both control and carrier donors. The sex ratio was similar in both centers and did not differ significantly from a 1:1 sex ratio. The larger chromosomes in the complement (1, 2, 3, 4, 5, 7, and 10) presented a significantly lower frequency of hypohaploidy, while some of the smaller chromosomes (13, 19, and 21) showed a higher frequency of hypohaploidy than expected. Chromosome 21 and the sex chromosomes showed an increase in the percentage of hyperhaploidy, as compared to other chromosomes, that was close to statistical significance (P = 0.08). Our results reflect a preferential loss of small chromosomes during slide preparation and suggest that chromosome 21 and the sex chromosomes could be more frequently involved in aneuploidy.

Aneuploidy↗

PubMed1995

An analysis of human sperm chromosome breakpoints.

Sperm chromosome analysis of 19 sperm donors with either normal or balanced karyotypes was carried out in order to explore the nature of sperm chromosome structural aberrations. A total of 2,389 cells (range 36-298/donor) were karyotyped after in vitro penetration of hamster eggs. The median percentage of sperm structural aberrations was 9.3% (SD +/- 4.7; range 0%-17.8%), with a total of 247 breakpoints, of which 220 could be characterized fully. Two sets of donors were studied in two different centers: center 1 (United States) and center 2 (Spain). The frequencies of nonrejoined and rejoined chromosome-type aberrations were very similar between center 1 and center 2: 83.6% and 10.0%, and 75.0% and 10.3%, respectively. Chromatid-type aberrations were more frequent in center 2 (14.7%) than in center 1 (6.4%) (P = .037). Chromosome 4 had less than the expected number of breakpoints (P < .001). A positive significant correlation was found between sperm breakpoints reported in this study and sites of balanced chromosome de novo rearrangements detected at prenatal diagnosis and reported in the literature (P = .0001).

Adult↗

PubMed1994

Inhibition by anti-sperm monoclonal antibodies of the penetration of zona-free hamster oocytes by human spermatozoa.

Fourteen mAb specific for human sperm membrane antigens were selected to investigate their inhibitory effect on fertilization. The antigens were not expressed in other human somatic tissues but were present in sperm from other species. The antibodies were purified from ascites fluid produced in mice. The zona-free hamster egg penetration assay was used for the evaluation of the blocking ability of these antibodies. The inhibition rate was generally related to a decrease in the number of adherent sperm. Three groups of antibodies were distinguished: (i) four mAb that have high inhibition at any concentration; (ii) four mAb with an intermediate inhibitory effect, that is more dependent on the antibody concentration tested; and (iii) six mAb with little or no effect at any concentration. The presence of antibodies leads to a lower penetration index, or number of penetrating sperm per oocyte. MAb specific for head antigens promote high inhibition of fertilization; these antibodies show a patchy staining on the sperm head. The antibodies localized in the midpiece have an intermediate inhibitory effect. No inhibition is detected with the equatorial region binding pattern. Sperm agglutination does not play any role in the inhibition caused by the antibodies described here.

Animals↗

PubMed1993

Segregation analysis in a man heterozygous for a pericentric inversion of chromosome 7 (p13;q36) by sperm chromosome studies.

We have analyzed 140 sperm chromosome complements from a subfertile man heterozygous for an inv(7)(p13;q36). Seventy-five percent of the chromosome complements were not recombinant: 37.9% contained the normal chromosome 7, and 37.1% contained the inverted chromosome 7. Twenty-five percent of the 140 were recombinant: 7.1% carried a recombinant chromosome 7 with a duplication p and deletion q, 17.1% carried a recombinant chromosome 7 with a duplication q and deletion p, and 0.7% carried both recombinant chromosomes. The frequency of structural chromosomal aberrations unrelated to the inversion was 11.4%, and the frequency of aneuploidy was 2.9%. Both frequencies were not significantly different from those in control donors. Two sperm complements with a second independent, contiguous inversion involving one of the original breakpoints (q36) were observed (1.4%). The risk of producing chromosomally abnormal offspring or spontaneous abortions would be 34.3%. The proportion of X-bearing and Y-bearing sperm was 46.8% and 53.2%, respectively, not significantly different from the expected 1:1 ratio.

Adult↗

PubMed1992

Cytogenetic studies in motile sperm from normal men.

Most studies on human sperm chromosomes from normal men involve the heterologous fertilization of zona free hamster eggs by unselected human sperm. In this work, we have performed cytogenetic studies of highly motile sperm, selected by a swim-up method. A total of 505 motile human sperm complements from three normal donors was analysed. The total frequency of sperm with chromosomal abnormalities (10.9%; 6.9% structural aberrations and 4.0% aneuploidy) and the sex ratio (50.4% X:49.6% Y) were similar to those obtained from whole semen samples. Our results indicate that the selection of motile sperm does not imply chromosomal selection.

Adult↗

PubMed1992

Repair of human sperm chromosome aberrations in the hamster egg.

In order to study the repair capacity of fertilized hamster eggs for the lesions present or induced in human sperm, we have examined the potentiating effect of caffeine, a DNA repair inhibitor, on the frequency and types of sperm chromosome aberrations. Sperm samples were donated by an individual treated with chemotherapy for a testicular cancer 3 years previously. Exposure of spermatozoa and inseminated oocytes to caffeine led to an increase of sperm chromosome aberrations, indicating that the damage to human sperm can be repaired in untreated hamster egg cytoplasm. The potentiating effect of caffeine was mainly reflected in an increase of unrejoined aberrations, indicating that the formation of chromosomal rearrangements is also inhibited. Since both chromatid-type and chromosome-type aberrations increase after treatment with caffeine, damage to human sperm can probably be repaired inside the hamster egg cytoplasm by pre- and post-replication repair mechanisms.

Animals↗

PubMed1991

Chromosome abnormalities in human spermatozoa after albumin or TEST-Yolk capacitation.

Cytogenetic studies were made on 328 spermatozoa from three individuals using either fresh semen samples capacitated in Biggers, Whitten and Whittingham (BWW) medium plus human serum albumin (BWW + HSA) or semen samples preserved at 4 degrees C in TEST-Yolk buffer. A total of 261 sperm karyotypes were obtained in a series of experiments in which half of each sample was capacitated in BWW + HSA and the other half in TEST-Yolk buffer at 4 degrees C for 2 days; 123 and 138 sperm karyotypes were obtained from the two capacitation methods respectively. Neither the frequency of sperm chromosomal abnormalities nor the sex ratio was significantly different after each capacitation methods. In one individual, however, the sex ratio (19X:32Y in the fresh sample and 49X:28Y in the preserved sample) did show a significant difference. In three experiments with semen samples from a single individual capacitated at 4 degrees C for 1, 2 or 3 days in TEST-Yolk buffer we obtained 33, 30 and 34 sperm karyotypes respectively. No significant differences in the sex ratio was exhibited between these experiments; the number of chromosome anomalies was too low to allow statistical analysis. Our results suggest that TEST-Yolk capacitation for 1, 2 or 3 days does not induce significant variations in the frequency and type of chromosomal abnormalities in human spermatozoa.

Animals↗

PubMed1991

XY-trivalent association and synaptic anomalies in a male carrier of a Robertsonian t(13;14) translocation.

Synaptonemal complexes were analysed in 91 pachytenes from a patient carrier of a Robertsonian translocation 45XY, t(13;14). Electron microscopic studies were carried out in 36 pachytenes. In some cases, sequential light microscopy/electron microscopic images were obtained (16/36). As the resolution of the light microscope does not allow an accurate characterization of anomalies, only EM images have been taken into account. Association of the trivalent and the sex vesicle was detected in 8 of 36 pachytenes (22.2%). Synaptic anomalies (25%), fragmentation of synaptonemal complexes (11.1%) or both (2.8%) were seen in 14 of 36 nuclei. According to previous studies, synaptic anomalies may produce a spermatogenic arrest. However, it has been suggested that synaptic anomalies may be the result of a process of cell degeneration, and not its cause. The non-random relationship between the sex vesicle and the translocation trivalent may result from the tendency of unpaired segments to associate with each other after homology search is relaxed and heterologous synaptic adjustment is allowed.

Adult↗

PubMed1990

Significance of structural chromosome aberrations in human sperm: analysis of induced aberrations.

A significant increase in the incidence of structural chromosome anomalies has been observed in the sperm of patients treated with radio and/or chemotherapy for different types of cancer when analyzed by the interspecific fertilization of hamster eggs. The analysis of these aberrations shows that while in controls only 9.4% of structural abnormalities are of the stable type, in treated patients this figure increases to 39.3%, thus indicating that the anomalies have not been produced during the fertilization of the hamster egg. However, it is possible that part, or even most, of the breaks appear as a result of a reduced repair capacity of sperm chromosomes in the cytoplasm of the hamster egg.

Antineoplastic Agents↗

PubMed1990

Lymphocyte and sperm chromosome studies in cancer-treated men.

To evaluate the reliability of the quantitative extrapolation of the long-term effect of cancer therapies from somatic cells to germ cells, we compared the frequency of chromosome abnormalities in 303 lymphocytes from four individuals treated with radio- and/or chemotherapy 5-18 years earlier with the frequency in 422 spermatozoa from the same individuals. The mean frequency of structurally abnormal complements was much higher in germ cells than in somatic cells (P = 2.08 x 10(-6)). The fact that spermatogenic cells share cytoplasm is suggested as a possible factor in the increased viability of germ cells with chromosome aberrations. In addition, in spermatozoa the incidence of structural chromosome abnormalities was much higher in treated individuals than in controls (P less than 0.00060), while in lymphocytes no statistically significant differences could be observed. This observation and the apparent lack of relationship between individual frequencies in the two kinds of cells suggest that the long-term effect of anti-tumor treatments on germ cells cannot be extrapolated from the analysis of somatic cells.

Antineoplastic Combined Chemotherapy Protocols↗