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J Egozcue

Publications and source records attributed to J Egozcue.

At least 73 records · Page 4Linked to original sources
PubMed2000

Preimplantation genetic diagnosis.

Preimplantation genetic diagnosis (PGD) includes a variety of techniques that have been developed to detect the transmission to the offspring of genetic diseases or of chromosome abnormalities by couples at risk before a pregnancy is established, to avoid these couples the risk of recurrent abortions and/or of repeated terminations of pregnancy. Candidate couples are carriers of gene mutations or of structural chromosome rearrangements, or with recurrent spontaneous abortions of unknown origin. Diagnostic procedures include different modalities of gene amplification using the polymerase chain reaction (PCR) or of fluorescent in situ hybridization (FISH). Embryo biopsies are carried out at the 6-8 cell stage. Healthy embryos are transferred on day 4 or at the blastocyst stage. By now, several hundred healthy children have been born using PGD, and only one diagnostic error has been reported.

Abortion, Habitual↗
PubMed2000

The decision to cancel a preimplantation genetic diagnosis cycle.

It has been suggested that a minimum number (six) of cumulus-oocyte complexes (COCs) should be retrieved for fertilization to offer enough chances to ensure a pregnancy after a preimplantation genetic diagnosis (PGD) procedure. Therefore a decision to cancel a PGD cycle should be adequately weighted to offer the patients the highest chances to obtain a pregnancy. We describe a case where, after retrieving only three COCs suitable for fertilization, a triplet pregnancy was obtained. This case suggests that, although low numbers of COCs can reduce the effectiveness of the PGD procedure, other factors are involved in its final result. Thus, the opportunity of routinely cancelling such cycles should be reconsidered. In addition, this is, to our knowledge, the first case where sex selection was carried out to prevent the birth of carriers of the abnormal gene, and not of affected offspring.

Adult↗
PubMed2000

Interchromosomal effects for chromosome 21 in carriers of structural chromosome reorganizations determined by fluorescence in situ hybridization on sperm nuclei.

We have used dual color fluorescence in situ hybridization (FISH) on decondensed sperm heads from four carriers of structural chromosome reorganizations, viz. t(3;15), t(Y;7), t(13;22) and inv(9), to assess the possible existence of an interchromosomal effect (ICE) on the segregation of chromosome 21. In the carriers of t(Y;7), t(13;22) and inv(9), all results were within the limits described in controls. A highly significant increase (P<0.0001) of disomy 21 (1.90% v 0.37%), which could be considered as a positive ICE, was observed in the t(3;15) carrier. Significantly higher percentages (P<0.0001) of diploid sperm (5.71% v. 0.27%) were also observed in this patient. Our results suggests that the occurrence of an ICE may depend on the reorganization and on the chromosome and chromosome regions involved, resulting in a particular meiotic behaviour (presence of unsynapsed regions, preferential meiotic configurations) that could lead to the observed increase in chromosome 21 disomies. Further studies with this technical approach in a wide range of structural reorganizations could help to elucidate the actual occurrence of ICEs.

Adult↗
PubMed2000

Cytogenetic characterization of two colon cell lines by using conventional G-banding, comparative genomic hybridization, and whole chromosome painting.

The heterogeneous nature of genetic alterations in cancer cells handicaps the full characterization of its occurrence and the analysis of their molecular bases and relation to biological processes. Although many cancer cells are highly aneuploid, in other cases, as in a subset of colorectal carcinomas displaying microsatellite instability, chromosomal aberrations are scarce. The aim of this study was to fully characterize both qualitatively and quantitatively, the karyotypes of two established colon carcinoma cell lines (LoVo and HCT 116) previously reported as being near diploid. An array of complementary cytogenetic techniques were used: G-banding, comparative genome hybridization (CGH), and whole-chromosome painting (WCP). Combinations of these techniques provided an accurate karyotype for the two cell lines: LoVo cells showed 49,XY,t(2;12)(q13;p11.2),+5,+7,+12,i(15)(q10) and HCT 116 cells showed 45,X,-Y,dup(10)(q24q26),der(16)t(8;16)(q13;p13), der(18)t(17;18)(q21;p11.3). Heterogeneity was also observed in both cell lines as shown by G-banding. Chromosomal unbalances determined by CGH (many of them related to structural reorganizations) were characterized by WCP, allowing the reliable identification of those chromosome markers that could not be completely identified by G-banding. We show that combined analysis with classical and molecular cytogenetic techniques provides an accurate map of chromosomal aberrations in these two cell lines not identified in previous investigations.

Adenocarcinoma↗
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↗
PubMed2000

Increased incidence of meiotic anomalies in oligoasthenozoospermic males preselected for intracytoplasmic sperm injection.

PURPOSE: Based on data from the literature, to detect the possible presence of an increased frequency of meiotic anomalies in oligoasthenozoospermic (OA) patients preselected for intracytoplasmic sperm injection. METHODS: Meiotic studies in as many successive patients with a clinical indication for a diagnostic testicular biopsy as needed to complete at least 100 cases with a severe OA (motile sperm concentration < or = 1.5 x 10(6)/ml). RESULTS: An increased incidence of meiotic anomalies was found in 102 patients with a severe OA (17.6%) compared to the mean for 105 patients with other etiologies in the series (5.7%) or the mean for patients reviewed in the literature (6.5%). CONCLUSIONS: Patients with a severe OA have a higher incidence of synaptic anomalies. This may result in the malsegregation of chromosomes at meiosis I, producing abnormal sperm, and could explain the high incidence of sterility and some cases of abortion (in two thirds of the couples with abortions the husband had meiotic anomalies) in this group.

Biopsy↗
PubMed2000

Effects of ionizing radiation on oocytes of prepubertally irradiated rats.

PURPOSE: To characterize the effects of ionizing radiation on the immature oocytes of prepubertal rats aged 16-20 days old. MATERIALS AND METHODS: The abdomen of Sprague Dawley (SD) female rats was exposed to 0, 1 and 2 Gy of X-rays. The effects were analysed when the animals became adults (80 days old) using as parameters the number of implantation sites, embryonic loss, dominant lethality and the synaptonemal complex (SC) analysis of F1 female foetuses. RESULTS: A statistically significant decrease in the fertility of the 2 Gy irradiated rats was observed. However, the SC analysis of F1 female foetuses showed no cytogenetic damage that could be attributed to radiation. CONCLUSIONS: A reproductive effect was demonstrated but no inherited genetic damage was detected by SC analysis in female foetuses of irradiated mothers.

Animals↗
PubMed2000

Is there a place for preimplantation genetic diagnosis screening in recurrent miscarriage patients?

Chromosomal abnormalities are one of the factors known to interfere with normal embryo development; thus preimplantation genetic diagnosis (PGD) for chromosome anomalies may be a new tool for improving the pregnancy rate in selected groups of patients. Embryos from three groups of patients (control, aged and recurrent miscarriage patients) were screened by PGD using specific DNA probes for chromosomes 13, 16, 18, 21, 22, X and Y. The control and aged groups were included in the PGD study because the women carried sex-linked genetic disease. The frequencies of chromosome anomalies observed in older women (46.3%) and in recurrent miscarriage patients (53%) were significantly higher (P < 0.05) than the frequency in the control group (19.3%). After PGD screening and transfer of normal embryos, pregnancies were obtained in women who had undergone repeated abortions (approximate 25% pregnancy rate per transfer) but not in older women. On the basis of these data, it seems that PGD screening of embryos can help some women undergoing repeated abortions, as these techniques allow successful pregnancies to be obtained or, if no pregnancy can be obtained, the results may help the couple to decide whether to enter an embryo or gamete donation programme.

Abortion, Habitual↗
PubMed1999

Study of allelic losses on 3p, 6q, and 17p in human urothelial cancer.

Forty-eight transitional cell carcinomas of the bladder and three transitional cell carcinomas of the renal pelvis were examined for loss of heterozygosity (LOH) on chromosomes 3p, 6q, and 17p. The most frequent allelic loss was seen on 17p (18/36, 50%) followed by 6q (6/22, 27%), and 3p (5/22, 23%). In cases with LOH at more than one locus, the same DNA sample often varied in degree of signal reduction for missing alleles. This observation indicates that LOH studies can serve to detect intratumor heterogeneity. No correlation was found between allelic losses at these chromosome arms and tumor grade and stage. Allelic losses on 6q were associated with tumors having a solid growth pattern; in this kind of tumors, allelic losses on 3p were associated with invasion.

Aged↗
PubMed1999

Chromosome instability in lymphocytes from two patients affected by three sequential primary cancers: the role of fragile sites.

The chromosomal aberration rate and the expression of fragile sites induced by aphidicolin were evaluated in metaphase chromosomes obtained from peripheral blood lymphocytes of two untreated patients with multiple primary cancers. Spontaneous aberrations of chromosome number and structure and chromosome fragility were compared with controls with the use of the same methods. Chromosomal aberration rates and expression frequencies of fragile sites were significantly higher in the patients than in normal control subjects. In the patients, all but one structural chromosome aberration involved at least one fragile site. Our results suggest that fragile sites may be unstable regions of the human genome, which might play an important role in the genetic instability associated with cancer predisposition.

Aged↗
PubMed1999

Characterization of constitutive heterochromatin in Cebus apella (Cebidae, Primates) and Pan troglodytes (Hominidae, Primates): comparison to human chromosomes.

Using G bands, some homologies between the chromosomes of Cebus apella (CAP) and human chromosomes are difficult to establish. To solve this problem, we analyzed these homologies by fluorescence in situ hybridization using human whole chromosome probes (ZOO-FISH). The results indicated that 1) the human probe for chromosome 2 partially hybridizes with CAP chromosomes 13 and 5, 2) the human probe for chromosome 3 partially hybridizes with CAP chromosomes 18 and 20, 3) the human probe for chromosome 9 partially hybridizes with CAP chromosome 19, and 4) the human probe for chromosome 14 hybridizes with the p-terminal and q-terminal regions of CAP chromosome 6. However, none of the human probes employed hybridized with the heterochromatic regions of CAP chromosomes. For this reason, we characterized the heterochromatic regions of CAP chromosomes and of the chromosomes of Pan troglodytes (PTR), to allow comparison between CAP, PTR, and human chromosomes using in situ digestion of fixed chromosomes with the restriction enzymes AluI, HaeIII, and RsaI and by fluorescent staining with DA/DAPI. The results show that 1) centromeric heterochromatin is heterogeneous in the three species studied and 2) noncentromeric heterochromatin is homogeneous within each of the three species, but is different for each species. Thus, centromeric heterochromatin undergoes a higher degree of variability than noncentromeric heterochromatin.

Animals↗
PubMed1999

Analysis of sex chromosome aneuploidy in sperm from fathers of Turner syndrome patients.

Numerical sex chromosome abnormalities were analyzed in sperm from four fathers of Turner syndrome patients of paternal origin to determine whether there was an increased frequency of sex chromosome aneuploidy and to elucidate whether meiotic malsegregation mechanisms could be involved in the origin of Turner syndrome. Determination of the parental origin of the single X chromosome (maternal in all four cases) and exclusion of X and Y mosaicism were carried out by polymerase chain reaction amplification of five X chromosome polymorphisms and three Y chromosome segments. A total of 45,299 sperm nuclei from Turner fathers and 85,423 sperm nuclei from eight control donors was analyzed by three-color fluorescence in situ hybridization. The four patients showed a significant increase in the percentages of XY sperm (mean 0.22%; range 0.20% to 0.22%) compared with control donors (mean 0.11%; range 0.06% to 0.18%). These results suggest that the four individuals have an increased frequency of nondisjunctional errors in meiosis I, resulting in the production of an increased proportion of XY spermatozoa and of sperm lacking a sex chromosome.

Adult↗
PubMed1999

In vitro fertilization plus preimplantation genetic diagnosis in patients with recurrent miscarriage: an analysis of chromosome abnormalities in human preimplantation embryos.

OBJECTIVE: To analyze the incidence of numeric chromosomal abnormalities in preimplantation embryos from women with unexplained recurrent miscarriage (RM) so as to seek an etiology and to determine whether the use of IVF may be indicated to treat these cases. DESIGN: Prospective controlled study. SETTING: University laboratory of reproductive genetics and a tertiary referral center for infertility. PATIENT(S): Nine women with a mean (+/-SD) of 3.9 +/- 0.6 RMs who were undergoing IVF and preimplantation genetic diagnosis, and a control group of young (n = 10) and older (n = 6) patients who were undergoing preimplantation genetic diagnosis because of sex-linked diseases. INTERVENTION(S): In vitro fertilization, embryo culture for 72 hours, blastomere biopsy, and analysis of chromosomes 13, 16, 18, 21, 22, X, and Y with the use of fluorescent in situ hybridization. Transfer of chromosomally normal embryos into the uterus. MAIN OUTCOME MEASURE(S): Numeric chromosomal abnormalities in human embryos. RESULT(S): Sixty-six embryos from patients with RM were compared with 62 embryos from young patients and 41 embryos from older patients. There was a significant increase in the rate of abnormal embryos in the patients with RM and the older patients compared with the controls. Abnormalities in most of the chromosomes studied were higher in the RM group than in the control group, especially those affecting chromosome 13. CONCLUSION(S): There was an increase in numeric chromosomal abnormalities in preimplantation embryos from women with RM that could be the cause of infertility in many couples with unexplained RM. The use of IVF in such circumstances may be indicated if successful preimplantation genetic diagnosis is added to the procedure.

Abortion, Habitual↗
PubMed1999

Screening for abnormalities of chromosomes X, Y, and 18 and for diploidy in spermatozoa from infertile men participating in an in vitro fertilization-intracytoplasmic sperm injection program.

OBJECTIVE: To evaluate the frequency of disomy (for chromosomes X, Y, and 18) and of diploidy in the spermatozoa of infertile men undergoing intracytoplasmic sperm injection (ICSI). DESIGN: Prospective analysis of sperm nuclei by fluorescence in situ hybridization (FISH). SETTING: University-affiliated IVF-ICSI program. PATIENT(S): Semen samples from 19 patients participating in an IVF-ICSI program. INTERVENTION(S): Semen samples were analyzed and prepared for FISH. MAIN OUTCOME MEASURE(S): Semen parameters were evaluated. The frequency of disomy for chromosomes X, Y, and 18 and the frequency of diploidy were analyzed by FISH. RESULT(S): A total of 9,373 spermatozoa from 19 infertile patients were analyzed and compared with spermatozoa from a control group of 5 healthy men. No differences in the frequency of disomy 18 were found, but statistically significant differences in the incidence of sex chromosome disomy and of diploidy were observed. CONCLUSION(S): The study of sperm nuclei by FISH is useful to improve genetic counseling in infertile patients selected for ICSI.

Adult↗
PubMed1999

Implications of sperm chromosome abnormalities in recurrent miscarriage.

PURPOSE: Our purpose was to assess the existence of sperm chromosome abnormalities in recurrent pregnancy loss in an assisted reproduction program. METHODS: In this prospective study, 12 sperm samples from couples undergoing in vitro fertilization with two or more first-trimester spontaneous abortions were analyzed. Diploidy and disomy in decondensed sperm nuclei were assessed for chromosomes 13, 18, 21, X, and Y using two- and three-color fluorescence in situ hybridization. RESULTS: Sex chromosome disomy in sperm samples from recurrent abortion couples was significantly increased compared to that from internal controls (0.84% vs 0.37%). In a subpopulation of seven couples who underwent oocyte donation, mean frequencies for sex chromosome disomy (1%) were even higher and diploidy (0.43%) was also significantly increased. CONCLUSIONS: These results suggest an implication of sperm chromosome abnormalities in some cases of recurrent pregnancy loss.

Abortion, Habitual↗
PubMed1999

Comparison of X-ray dose-response curves obtained by chromosome painting using conventional and PAINT nomenclatures.

PURPOSE: The compare the suitability of PAINT and conventional nomenclature systems for the construction of chromosome aberration dose-effect curves for X-rays using FISH techniques, and to compare these curves with those based on solid-stained dicentrics analysed in first division metaphases by the FPG technique. MATERIALS AND METHODS: Blood samples were irradiated at 0.1, 0.25, 0.50, 0.75, 1, 1.5, 2, 3, 4 and 5 Gy 180 kV X-rays. FISH painting was performed using probes for chromosomes 1, 4 and 11 in combination with a pan-centromeric probe. RESULTS: Translocations showed a higher background frequency than dicentrics. This influences the ratio of translocations:dicentrics at the lower doses and the uncertainties of dose-effect curves for translocations. The dose-effect curves for dicentrics obtained by FISH and solid stain were in close agreement. CONCLUSION: For short-term biological dosimetry purposes by FISH, the use of dic(BA) (PAINT nomenclature) or total dicentrics (conventional nomenclature) should give similar dose estimates. For dose reconstruction, the use of total or complete translocations result in similar uncertainties.

Adult↗
PubMed1999

Effect of 6 beta-methylprednisolone on mouse pregnancy rate.

The aim of this study was to evaluate the effect of methyl-prednisolone on the pregnancy rate in mice. For this reason, zona pellucida-intact and zona pellucida-free embryos at the blastocyst stage were transferred to recipient mice at day 2.5 of pseudopregnancy. Embryo transfer was performed into non-immunodepressed and immunodepressed groups of recipient mice using 0.3 or 0.6 microgram/g of 6 beta-methylprednisolone. A higher implantation and developmental rate of zona-free embryos transferred to the immunodepressed group of recipients was observed after using the higher dose of methylprednisolone.

Animals↗