Modern techniques of sperm preparation--do they influence the sex of offspring?

Four different semen preparation methods--migration-Sedimentation technique, Sperm Select, Percoll gradient centrifugation and swim-up technique--have been tested and compared in regard to progressive motility, viability and morphology. The data obtained clearly demonstrate significant enhancement of all parameters using the techniques mentioned above. The study also demonstrated significant differences in Y chromatin-positive spermatozoa before and after semen preparation. The rate of F-body-positive spermatozoa was significantly increased by all four methods.

Post-transfusion graft-versus-host disease following open heart surgery. Report of six cases.

Six cases of post-operative erythroderma after open heart surgery are described. About 10 days after seemingly uneventful recovery, all patients developed fever, erythroderma, liver enzyme elevation, pancytopenia, and an aplastic bone marrow. Their condition rapidly deteriorated, and they died within 20 days of the onset of symptoms. Skin biopsy specimens from two patients showed mild leukocytic infiltration in the epidermal basal layer and upper dermis. Immunostaining by the ABC method showed that most of these infiltrating cells were suppressor/cytotoxic T cells. HLA study of peripheral lymphocytes from two patients and their families revealed that the patients' HLA phenotypes were incompatible from their children's HLA findings. Y chromatin was present in the lymphocytes of the skin biopsy specimen of a female patient. Based on the clinical picture, skin biopsy, HLA study, and Y chromatin study, the authors strongly suspect post-transfusion GVHD as the etiology of postoperative erythroderma, although these patients lacked any known immunodeficiency.

Síndrome cromossômicas detectadas em rotina laboratorial através dos exames de cariótipo e de cromatina sexual X de Barr / Chromossomics syndromes detected in the laboratory routines with cariotype and sexual X of Barr cromatine exams

Os autores realizaram, na rotina do Laboratório Oswaldo cRuz (S.P.), exames de cariótipo e de cromatina sexual X de Barr em 120 pacientes. Destes, 26 (21,66%) dos pacientes apresentaram síndromes derivadas de anomalias cromossômicas numéricas e estruturais

A DNA probe from Schistosoma mansoni allows rapid determination of the sex of larval parasites.

A DNA clone representing a 0.4 kb degenerative repeat has been isolated. The DNA sequence is present only in the genome of female Schistosoma mansoni at different stages of the life cycle, at a frequency of approximately 75 copies per adult female genome. The sequence is not expressed and probably represents satellite DNA in the heterochromatin region of the W chromosome. It is demonstrated that the DNA clone may be used for the rapid determination of the sex of cercariae without the need for DNA isolation or Southern blotting.

[Studies on sex determination using human dental pulp. II. Sex determination of teeth left in a room].

The appearance rate of Y chromatin was calculated in the dental pulp with the staining by Quinacrine Mustard. The morphological findings of the tissue were observed with the HE stained section and the change of the appearance rate of Y chromatin was studied along with the morphological change of the tissue. The average of the appearance rate of Y chromatin in the male dental pulp was 42.2% immediately after the tooth extraction, 34.8% after one month, 27.9% after 3 months, 20.8% after 6 months, 20.6% after 1 year, 19.1% after 18 months and 11.4% after 2 years, respectively. On the other hand, the appearance rate of the pseudo-Y chromatin spot in the female pulp was below 4.2% through the two years' observation which enabled to determine the sexuality. By the HE staining observation about the dental pulp, left at a room temperature, it was revealed that the pulp tissue can be well preserved for a long time because of the natural drying. The degree of the staining declined with the lapse of time after the removal of tooth. Degeneration of tissue did not offer any discrete criteria for the estimation of the lapse of time after the removal of tooth.

Síndromes cromossômicas detectadas em rotina laboratorial através dos exames de cariótipo e de cromatina sexual X de Barr

Os autores realizaram, na rotina do Laboratório Oswaldo Cruz (S.P.), exames de cariótipo e de cromatina sexual X de Barr em 120 pacientes. Destes, 26 (21,66%) dos pacientes apresentaram síndromes derivadas de anomalias cromossômicas numéricas e estruturais

Femininity control at the XXth Universiade in Kobe, Japan.

In 1985, we were in charge of the femininity control tests for the participants of the XXth Universiade in Kobe. At that time, we chose as screening tests checking both X chromatin from buccal mucous membrane and Y chromatin from hair roots. The number of female competitors who attended the Universiade, except tennis, gymnastics, and fencing, was 631. Among them, 108 had already had the certificate of femininity so we examined 523 female competitors. All but three of them were normal for femininity, i.e., X chromatin positive and Y chromatin negative. Two of the three exceptions were Y chromatin positive and X chromatin negative. Chromosomal analyses from their peripheral lymphocytes showed a pattern concordant with a 46,XY karyotype. The other was both X and Y chromatin positive, but we could find Y chromatin on only one slide out of three. Since this person refused to have a blood sample taken, we can only suppose her karyotype as being 46,XX/46,XY or 46,XX/47,XXY. This last case would not have been found, if we had checked only the X chromatin. From our experience, we conclude that for femininity control, it is safer to screen the Y chromatin. We would strongly advise the use of both X and Y chromatin tests for femininity control.

A simple technique for sex chromatin analysis in amniotic cells of mouse and rat embryos.

A simple technique has been developed for sex chromatin analysis in the amniotic membrane of rodent embryos. This technique combines the use of 60% acetic acid as a fixative and a carbol fuchsin stain. This technique may be useful for quick sex diagnosis of rodent embryos at midgestation for various purposes in experimental embryology.

Papel das alteraçöes cromossômicas na etiologia da esterilidade masculina / Role of chromosome changes in the etiology of male sterility

Anomalias cromossômicas no homem podem levar à esterilidade. O estudo citogenético de 40 pacientes com histórico de azoospermia ou oligospermia severa foi realizado visando estimar a freqüência de anomalias cromossômicas na etiologia da esterilidade primária e avaliar se esse exame deve ser recomendado como subsídio para o diagnóstico em nosso meio. A constituiçäo cromossômica normal 46,XY estava presente em 31 pacientes; quatro eram portadores de anomalias cromossômicas: cariótipo 47,XXY; 47,XYY; 45,XY,t (13q14q); homem XX; três apresentavam instabilidade cromossômica e dois a variante cromossômica Gp+. A etiologia da esterilidade primária foi determinada em 10% dos pacientes. Discutiu-se o papel das instabilidades cromossômicas e das variantes cromossômicas. O estudo citogenéticos revelou-se relevante como subsídio ao diagnóstico

Hypothyroidism and sex chromosomes.

The observation of Campbell and Price in 1979 that their Unit had diagnosed four subjects with both Klinefelter's syndrome and congenital hypothyroidism raised the suspicion of an association between the two conditions. This, and the published reports of an XX male, five XXY males, and one mosaic XY/XXY with congenital or acquired forms of hypothyroidism, together with the higher incidence in women and the absence of sex difference among inherited congenital cases, suggested a possible sex chromosome effect in the aetiology of sporadic hypothyroidism. Various hypotheses can be tested either by examining the frequency of hypothyroidism in sex chromatin positive males or by establishing a higher frequency of sex chromatin positive males among hypothyroid cases than in normal males. We examined 57 boys with hypothyroidism for the presence of sex chromatin and found all to be negative. From this relatively small sample we can only exclude the possibility of a very large (100 fold) difference in frequency between the two populations and therefore more data are needed.

[Müllerian adenosarcoma of the uterus]. / Miullerovskie adenosarkomy matki.

Histologic structure of Mueller's adenosarcoma has been studied in 5 patients subjected to uterine and its appendages extirpation. Microspectrophotometric assessment of the course of nuclear DNA accumulation, measurement of X-chromatin in two morphologic components of the tumor (epithelial and stromal) confirmed malignant morphological nature of the latter and benign one of the former. Basing on their experience with the 5 patients three of which died in the course of 5 years of relevant metastases, the authors believe the prognosis for uterine adenosarcoma to be poor.

[The 48,XXXX syndrome: study of psychomotor development from birth to 11 years of age and review of the literature]. / Syndrome 48,XXXX: étude du développement psychomoteur de la naissance à 11 ans et revue de la littérature.

A child with four X chromosomes is described. This case and the literature review allow to underline the mental retardation and some other "major" but inconstant signs that are extremely helpful for the early clinical diagnosis. They are hypertelorism, epicanthal fold and genital anomalies. The mental evolution is assessed on an eleven year period. The bad results concern particularly the child's use of language and the complicated works. They become worse with time. The additional X chromosomes Mary Lyon inactivation, perhaps incomplete, is discussed because its determinism. Enzymatic measuring out is our approach to this problem.

Sex chromatin analysis of lymphocytes invading host organs in transfusion-associated graft-versus-host disease.

We report a method of sex chromatin analysis of lymphocytes separated from host organs in transfusion-associated graft-versus-host disease (GVHD), which enabled the demonstration of invasion by donor lymphocytes. The lymphocytes examined were separated from deparaffinized tissue blocks of skin, spleen and bone marrow from two female patients with transfusion-associated GVHD by incubation in 0.5% pepsin. The tissues had been removed at autopsy and fixed in formalin. Sex chromatin analysis was performed by fluorescence microscopy on separated lymphocytes stained with 0.005% quinacrine dihydrochloride. By this means Y chromatin-positive (i.e. male) lymphocytes were demonstrated in the skin, spleen and bone marrow of both female patients.

Lymphoid chimerism after allogeneic bone marrow transplantation. Y-chromatin staining of peripheral T and B lymphocytes and allotyping of serum immunoglobulins.

Lymphoid cell engraftment was monitored for several years after bone marrow transplantation by Y-chromatin staining of T and B lymphocytes in the peripheral blood and/or by immunoglobulin allotyping in the serum of 20 of 52 pediatric patients grafted successively between October 1973 and October 1983. Data on 2 patients with severe combined immunodeficiency, grafted earlier in December 1968 and April 1971, are also included. These children received an allogeneic bone marrow graft for leukemia (n = 7), severe aplastic anemia (n = 11), or severe combined immunodeficiency (n = 4) and were informative for this study, because they differed from their donor by sex (n = 16) and/or by immunoglobulin phenotype (n = 13). Of 16 pairs in which the donor was of the opposite sex, 11 patients ultimately showed circulating T and B lymphocytes of donor origin after bone marrow transplantation; in the remaining 5, there was an incomplete chimerism of the circulating lymphoid cells. Of 13 pairs with a difference in immunoglobulin phenotype between donor and recipient, 8 patients exhibited donor allotypes 3 months or later after transplantation, in 3 of them together with recipient allotypes. In the remaining 5 patients, recipient allotypes were detected after transplantation, but the simultaneous presence of donor-type immunoglobulin production could not be excluded in 4. The persistence of either a split (T lineage of donor origin and B lineage of recipient origin) or mixed (T and/or B lineage of donor and recipient origin) chimerism was related to the type of disease. In 3 children circulating B cells of donor-origin did not fit with the recipient origin of the sessile immunoglobulin-secreting plasma cells. This implies that different immune compartments--e.g., bone marrow and peripheral lymphoid tissues--should be investigated following allogeneic bone marrow transplantation. A prolonged presence of recipient-type lymphoid cells increased the risk of leukemic relapse in the patients investigated.

[Levels of DNA and sex chromatin bodies in the myocyte nuclei of different sections of the human heart]. / Soderzhanie DNK i telets polovogo khromatina v iadrakh miotsitov raznykh otdelov serdtsa cheloveka.

Nuclei of ventricular, atrial and atrioventricular node myocytes of normal and hypertrophied human heart were studied on squash preparations and on 12 micron sections after the Feulgen staining. The cytophotometric DNA measurements have shown a distinction in the degree of polyploidization of nuclei in different heart compartments. In contrast to ventricular and atrial myocardia, in which polyploid nuclei predominate, the conduction system myocytes contain 77-88% of diploid nuclei. A correlation between DNA content and the number of sex chromatin bodies was observed for myocyte nuclei from all the compartments under investigation.