Chromosomal abnormalities and polymorphisms in Mexican infertile men.

A cross-sectional study was conducted to estimate the prevalence of chromosome abnormalities and normal variable chromosome features (polymorphisms) in infertile men from northeastern Mexico. Karyotyping was carried out in 326 men with diagnosis of infertility. The sperm counts showed 204 patients with oligozoospermia, 87 with azoospermia and 35 normozoospermia. Five patients with oligozoospemia and two with azoospermia presented chromosome abnormalities. Nonzoospermic men did not show chromosomal abnormalities. Polymorphisms of heterochromatin and satellite length showed a significant increased in oligozoospermic and azoospermic men with respect to normozoospermic men, respectively. This study reports the prevalence of chromosome abnormalities, polymorphisms of heterochromatin length, and polymorphisms in satellites in Mexican infertile men. The prevalence in this study was similar to other studies in world literature.

Loss of restriction site DdeI, used for avian molecular sexing, in Oreophasis derbianus.

Molecular sexing is a rapid and safe procedure for bird sex determination. Two universal methods based on the amplification of a chromo-helicase-DNA-Binding 1 (CHD) gene region, located in both sexual chromosomes (Z and W), have been established. We found that molecular sexing of Oreophasis derbianus failed by using these two procedures. One of them is based on a restriction site located in CHD1W gene but absent in CHD1Z. The DdeI restriction site, used successfully to determine gender in several bird species, was found to be lost because of nucleotide change in O. derbianus. This change created a new restriction site, NlaIII, that was successfully applied for sexing this endangered bird.

Polymorphism of Ag(+)-NORs in cervical smears from women with cervical cancer.

OBJECTIVE: To evaluate Ag(+)-stained (Ag(+)-NOR) polymorphism in four groups of patients with various grades of cervical lesions and in a control group. STUDY DESIGN: Forty-five women were selected, diagnosed and classified on the bases of the Pap smear and colposcopy/biopsy at Hospital de Ginecologia y Obstetricia del Instituto Mexicano del Seguro Social in Monterrey, Mexico. Five categories were considered: (1) inflammatory, (2) low grade squamous intraepithelial lesions (LSILs), (3) high grade squamous intraepithelial lesions (HSILs), (4) invasive cervical cancer, and (5) normal. The cervical smears were stained by the Ag(+)-NOR method. One hundred cells per slide were counted and classified according to the polymorphism of Ag(+)-NOR dots: typical (spherical) and atypical (large, kidney shaped and clustered). The four shapes of Ag(+)-NORs were quantified by percentage and transformed using the arcsine root procedure. RESULTS: Statistical analysis showed a significant decrease in spherical shape according to neoplastic development. The three atypical shapes showed a significant increase in patients with HSIL and invasive carcinoma in respect to LSIL. Principal components analysis grouped the data at five locations in the plane formed by the first two principal components according to the diagnosis. CONCLUSION: These findings suggest the potential diagnostic and prognostic value of the determination of Ag(+)-NOR polymorphism in cervical cytology studies.

Cytological effects of 60 Hz magnetic fields on human lymphocytes in vitro: sister-chromatid exchanges, cell kinetics and mitotic rate.

Incubation for 72 h of human peripheral blood cultures in the presence of 60 Hz sinusoidal magnetic fields (MF) at magnetic flux densities of 1.0, 1.5, and 2.0 mT led to stimulation of lymphocyte proliferation but had no influence on the frequency of sister-chromatid exchanges (SCE). The cytotoxic potential of MF combined with the mutagen Mitomycin-C also was analyzed. An opposite effect between MF exposure and Mitomycin-C treatment in terms of cell kinetics and mitotic rate was found, whereas no variation in SCE frequency was observed for this coexposure condition.

Sister chromatid exchanges in peripheral lymphocytes from women with carcinoma of the uterine cervix.

Sister chromatid exchanges (SCE) are reciprocal exchanges between sister chromatids. It has been reported that in patients with cervical cancer, the frequency of SCE in peripheral lymphocytes is significantly higher than that in normal individuals; however, other studies have shown no significant difference. The aim of this unmatched case-control study was to compare the mean number of SCE per metaphase in lymphocytes from women with and without carcinoma of the cervix uteri. The SCE specimens were prepared by the fluorescence plus giemsa technique in peripheral lymphocytes from 28 women with carcinoma of cervix uteri and 28 controls. The mean number of SCE per metaphase in women with carcinoma of cervix uteri (7.80 +/- 1.05) was higher than the control group (6.98 +/- 1.13) (P < 0.05; t-test). This study had a statistical power of 0.80 and an alpha value of 0.05. This finding suggests that an increased number of SCE in peripheral lymphocytes is associated with cervical cancer. We consider that the lack of reported association of SCE and cervical cancer might be attributed to the none determination of the statistical power and sample size.

Gonadal dysgenesis and Rokitansky syndrome. A case report.

BACKGROUND: Primary amenorrhea and lack of sexual development occur in gonadal dysgenesis due to missing ovaries. Primary amenorrhea with sexual development occurs in Rokitansky syndrome due to absence of the uterus, with normal ovarian function. The association of these two conditions has been previously described as a rare event. CASE: A 19-year-old woman presented with primary amenorrhea and lack of secondary sexual characteristics. Physical examination confirmed the absence of mammary development and of pubic and axillary hair. Pelvic ultrasound disclosed absence of the uterus and ovaries. Gonadotropin serum levels were in the menopausal range, and the karyotype showed two mosaic cell lines, 45,X/46,Xdic(X). Scanning of a large number of cells by interphase fluorescence in situ hybridization showed 12% of cells with a dicentric X chromosome. Laparoscopic study confirmed the absence of the uterus and ovaries, with normal fallopian tubes. CONCLUSION: This patient had two anomalies affecting reproductive performance, gonadal dysgenesis and congenital absence of the uterus, the first associated with an abnormal karyotype; the second seems to have occurred coincidentally. At this time there is no treatment for the reproductive dysfunction.

Mutagenic activity of diazepam evaluated by in vivo cytogenetic tests.

BACKGROUND: Diazepam, one of the benzodiazepine group of tranquilizers, is used as an adjunctive drug for sedation and for relief of anxiety in the treatment of epilepsy. Suspicion has been aroused of a possible mutagenic and teratogenic effect of this drug, thus the potential for cancer development. METHODS: To analyze the mutagenic effect of diazepam, the micronuclei and sister chromatid exchange (SCE) tests were performed by in vivo techniques in the bone marrow of Balb-C mice after intraperitoneal drug administration. Sixty mice, 30 males and 30 females, were classified as negative control (n = 12), positive control (n = 12), and three groups were treated with diazepam (n = 36). All groups were matched by sex, and each mouse received a single intraperitoneal injection. Negative control group was injected with physiological saline, positive control group with mitomycin-C at a dose of 0.5 mg/kg of body weight. Treated groups received diazepam, one at 0.1, the other at 0.2, and the last, at 0.4 mg/kg. RESULTS: The results showed a significant increase in the frequency of micronucleated polychromatic erythrocytes at all doses tested for whole population in relation to negative control. The polychromatic/normochromatic erythrocyte ratio showed a significant decrease at doses of 0.1 and 0.4 mg/kg in relation to negative control. The frequency of SCE was significantly higher at doses of 0.2 and 0.4 mg/kg in relation to negative control, the male mice being those affected. CONCLUSIONS: It is concluded that diazepam showed mutagenic and genotoxic effects on bone marrow cells of mice and that it might represent a human health risk.

Activity satellite association and polymorphism of Ag stained nucleolus organizer regions (Ag+ NORs) in lymphocytes from women with cervical uterine cancer.

Thirty five female patients with different stages of neoplastic lesions: cervical intraepithelial neoplasia (CIN) or dysplasia (CIN I and CIN II), in situ carcinoma (CIS), and adenocarcinoma, and 27 healthy women (controls) were studied to determine the activity, satellite association, and polymorphism of Ag stained nucleolus organizer regions (Ag+ NORs) in acrocentric chromosomes in metaphases obtained from peripheral blood lymphocytes. For each person, 25 to 50 metaphases stained with ammoniacal silver technique were scored. The average number of Ag+ NORs was higher in women with adenocarcinoma (7.66 +/- 0.72) than in controls (6.65 +/- 0.74). Non-associated chromosomes showing Ag+ NORs were found more frequently in patients (5.85 +/- 0.88) than in controls (4.81 +/- 0.67). Patients aged 30-39 and 60 or more had an increase of Ag+ NORs (7.99 +/- 1.04, and 7.81 +/- 0.71) with respect to their controls (6.36 +/- 0.052 and 6.17 +/- 0.88), but the frequency of satellite association showed lower values in 50-59 year-old patients (0.75 +/- 0.08) than in controls (1.02 +/- 0.19). The most frequent association in patients was the large type (patients = 38.96%, controls = 30.49%). The partial association showed higher values (6.49%) than controls (2.44%). Otherwise, the spherical association was more frequent for controls (37.80%) than for patients (28.57%). All these differences were statistically significant (p < 0.05). The frequency of Ag+ NORs and the type of polymorphism of satellite association could be related to the neoplastic process, while the frequency of satellite association and of polymorphism of Ag+ NORs seems to be irrelevant.

A case of mosaic Down's syndrome with two Robertsonian translocations.

A baby with stigmata of Down's syndrome was found to be a mosaic with two different cell lines: 45,XX,der(14q;21q)/46,XX,der(21q;21q)+21. The chromosome rearrangements appeared to have risen de novo. Four mechanisms are discussed for the origin of the mosaicism: dissociation of a translocation (14q;21q) chromosome already present in the 45,XX, der(14q;21q) zygote; two translocation events occurring sequentially at the first and second zygote (46,XX) divisions; a chromatid translocation in a 47,XX,+21 zygote; and an independent origin of the two cell lines.

Genetic structure of the populations migrating from San Luis Potosi and Zacatecas to Nuevo León in Mexico.

The Mexicans residing in the Monterrey metropolitan area in Nuevo León, Mexico, were grouped by generation and birthplace [Monterrey Metropolitan Area (MMA), San Luis Potosi (SLP), and Zacatecas (ZAC)] of the four grandparents to determine the extent of genetic variation within this population and the genetic differences, if any, between the natives living in the MMA and the immigrant populations from SLP and ZAC. Nine genetic marker systems were analyzed. The genetic distance analysis indicates that SLP and ZAC are similar to the MMA, irrespective of birthplace and generation. Gene diversity analysis (GST) suggests that more than 96% of the total gene diversity (HT) can be attributed to individual variation within the population. The genetic admixture analysis suggests that the Mexicans of the MMA, SLP, and ZAC, stratified by birthplace and generation, have received a predominantly Spanish contribution (78.5%), followed by a Mexican Indian contribution (21.5%). Similarly, admixture analysis, conducted on the population of Nuevo León and stratified by generation, indicates a substantial contribution from the MMA (64.6%), followed by ZAC (22.1%) and SLP (13.3%). Finally, we demonstrate that there is no nonrandom association of alleles among the genetic marker systems (i.e., no evidence of gametic disequilibrium) despite the Mestizo origin of this population.

[Frequency of color blindness and glucose-6-phosphate dehydrogenase enzyme deficiency in non-industrialized populations in the state of Nuevo León, México]. / Frecuencia de la ceguera para los colores y de la deficiencia a la enzima glucosa-6-fosfato-deshidrogenasa en poblaciones no industrializadas del Estado de Nuevo León, México.

In order to know if there would be genetic structural differences among non industrial and industrial populations, two genetic markers were studied: color-blindness (CPC) and glucose-6-phosphate dehydrogenase deficiency (G6PD), in students, males and females that were resident in five non industrial populations in the State of Nuevo Leon. The results were compared with the information for industrial zone from the Monterrey Metropolitan area (AMM). It was found that the frequencies of CPC and G6PD in non industrial populations (2.57 and 0.00 per cent), were lower than the ones in the industrial AMM (4.0 and 0.66 per cent), probably as a result that in the first populations, with minor urbanization, the main factors that influence are: natural selection, interacial mixed or genetic drift and the second population is the immigration, since 1940 to present time, of Mexican populations with greater influence from the Indians and Africans.

[Frequency of sister chromatid exchange in mice treated with gossypol]. / Frecuencia de intercambios de cromatides hermanas (ICH), en ratones tratados con gosipol.

To evaluate the possible mutagenic activity of gossypol, -a male contraceptive , which besides has inhibitory action on some parasites growth. Balb-C mouse received intraesophageally 10, 20 and 30 mg/Kg of gossypol to observe the effect of this drug on the frequency of sister chromatid exchanges (SCE) in bone marrow cells. A dose-response relation was not observed, and a small increase in the frequency of SCE was obtained. As it was not the double among the negative control, the gossypol effect must be classified as "undetermined" or "provisionally positive" agent. It is important to make more studies to determinate its possible mutagenic potential on humans.

[Activity of ribosomal genes and the association with acrocentric chromosomes in trisomy 21 and habitual abortion]. / Actividad de genes ribosomales y asociación de cromosomas acrocéntricos en la trisomía 21 y el aborto habitual.

The frequency of silver stain of the nucleolus organizer regions (NOR's) an indicator of the transcriptional activity of the DNAr, and the frequency of the association between acrocentric chromosomes (A.A.C.), possible indicator of the non-disjunction risk, were studied in the chromosomes in metaphase of eight married couples with regular 21 trisomy children, five couples that have had consecutive miscarriages and fifteen control normal persons, eight men and seven women. In this three groups the mean age was 30 years and they were compared according to sex. Another seven couples, with a mean age of 52 years also parents of 21 trisomic were compared against the group of young parents of 21 trisomics in order to analyze the age effect. Moreover there were studied the trisomic children of eight of the above couples in order to find out the effect of the supernumerary 21 chromosome. In all the groups the correlation coefficients between the frequency of NOR's-Ag+ and the A.A.C. were determined. It was found that the young group of parents of trisomic and the couples with consecutive miscarriages, both men and women, had a significant increased frequencies of NOR's-Ag+ and A.A.C. in comparison to controls. In the parents of trisomic, either men or women it was found a significant diminished level in the frequency of NOR's-Ag+ when the age increased, meanwhile raise in the frequency of A.A.C. with the increase of age was observed only in the women. A positive correlation between the frequencies of NORs-Ag+ and A.A.C. was found in the women of the control group and in the young mothers of trisomic.(ABSTRACT TRUNCATED AT 250 WORDS)

Low-speed purification of human placental nuclei.

A simple method for the purification of human placental nuclei is described. Nuclei were isolated by homogenizing tissue in standard saline citrate solution in the presence of zinc chloride to stabilize the nuclear membranes, NP40 as non-ionic detergent and sodium bisulphite for inhibition of proteolytic activity. Nuclei purification was achieved by low-speed centrifugation through a discontinuous sucrose gradient. The purified nuclei were evaluated by morphological criteria using phase contrast and electron microscopy. The extent of contamination by cytoplasmic debris was estimated by Papanicolaou's staining technique. Biochemical criteria include measurements of alkaline phosphatase activity as a plasma membrane enzyme marker and DNA-dependent RNA polymerase activity for the functional integrity of nuclear components. Transcriptionally active nuclei were obtained but the yield of nuclei was low; however, this low yield is compensated by the high degree of purity, the simplicity of the method and the functional and morphological integrity of the purified nuclei.

The effect of diphenylhydantoin on the frequency of micronuclei in bone-marrow polychromatic erythrocytes of mice.

Using the micronucleus test to evaluate the mutagenic effect of 5,5-diphenylhydantoin (DPH) on bone marrow polychromatic erythrocytes, male Balb-C mice were treated with the drug in single and multiple injection tests. A significant increase in the frequency of micronucleated polychromatic erythrocytes (MPE), P less than 0.05, was found when the mice received a single injection of DPH at doses of 0.5 and 1.0 mg/kg, and this frequency did not increase at higher doses. When mice were treated 3 times, at 24-h intervals, with 1.0 mg/kg of DPH, a significant increase in MPE was also observed (P less than 0.05) but this was lower than when they received a single injection of the same dose. A cytotoxic effect of NaOH, 0.1 N, which was used as solvent, was also observed either when alone or when DPH (1.0 mg/kg) was injected 3 times. This effect was comparable to the one produced by mitomycin C (MMC) at a dose of 0.5 mg/kg.