Apoptosis in spermatozoa of infertile men, clinical correlations

The methods for evaluation of male infertility include not only routine investigations, standardized by the WHO, but also complementary techniques, developed over the last years, in order to improve the predictive value of seminal analysis for natural conception and assisted reproduction. With reference to these new methods, studies suggest that sperm with certain levels of DNA fragmentation serve as a strong predictor of reduced male fertility. We studied subjects who underwent seminal fluid evaluation, because of an infertility condition, at the Department of Biomedical Sciences of the University of Sassari.The samples collected by masturbation were evaluated according to the World Health Organisation (1999).The samples was washed twice in PBS and cytocentrifuged for 10 min at 1800 rpm on polylysine-coated slides that were fixed in methanol at room temperature. The apoptosis was evaluated using the TUNEL (In Situ Cell Death Detection Kit, Fluorescein, Roche, Cat.No. 1 684 795). At fluorescent miscroscopy are counted at least 300 cells. Quantitative evaluation of apoptosis by the TUNEL method confirmed that apoptosis did not seem to be correlated with sperm concentration or morphology; however, we found a higher apoptotic rate in semen from patients affected by andrologic diseases, such as varicocele, than from those with alteration of semen characteristics. Apoptosis analysis might be used in infertile patients in order to understand the etiology of unexplained infertility and to improve therapeutic effectiveness

Unbalanced rearrangement der(9;18)(p10;q10) and JAK2 V617F mutation in a patient with AML following post-polycythemic myelofibrosis

Polycythemia Vera (PV) is a clonal myeloproliferative disorder characterized by excessive erythrocyte production, which may evolve into myelofibrosis and acute myeloid leukemia. Transformation to myelofibrosis occurs in 15-20% of cases and leukemic transformation in 5-10% of patients. The median survival time is 8-11 years and the median age at diagnosis is over 60 years. Normal karyotype is present at diagnosis in the majority of patients, while during transformation several acquired chromosome anomalies are present as trisomy 9 and gains in 9p

Effects of Cadmium chloride on human fetal cells in vitro

The principal aim of this work was to demonstrate the feasibility of tests with substances known as teratogenic in vivo on cell types which are the real target of their teratogenic effects. To this purpose Cadmium chloride has been tested on human amniotic fluid cells using the Chromosome aberrations (CA) and Sister chromatid exchanges (SCE) tests

Presence of telomeric and subtelomeric sequences at the fusion points of ring chromosomes indicates that the ring syndrome is caused by ring instability

In situ hybridization of a telomeric (TTA-GGG)n sequence to metaphases from three cases of ring chromosome, involving respectively chromosomes 4, 16, and 20, showed the presence of the cognate sequences in all three rings. To investigate whether these ring chromosomes originated by telomere-telomere fusion, we determined, by in situ hybridization, whether telomere-associated sequences and/or specific distal sequences were still present in the ring chromosomes. The finding that these sequences were preserved in all the ring chromosomes strongly indicates that they originated by telomere-telomere fusion. All three subjects carrying the ring chromosomes are affected by the so-called ring syndrome, with failure to thrive, minor dysmorphic signs and no major anomalies. The r(4) patient has the ring in mosaic form with a normal cell line and has normal intelligence. The r(16) and the r(20) patients have moderate mental retardation and suffer from seizures. We conclude that the ring syndrome, even in its more severe manifestation, is caused by ring chromosome instability

A Novel autosomal dominant non-syndromic deafness locus (DFNA48) maps to 12q13-q14 in a large Italian family

Non-syndromic hearing loss is the most common sensory disorder in humans; 15%–20% of cases are transmitted as a dominant trait (NSDA) with 40 loci having been mapped and 16 genes having been identified. Here, we report the mapping of a novel NSDA locus, DFNA48, to chromosome 12q13-q14 in a large multigenerational Italian family. A maximum lod score of 3.31 was obtained with marker D12S83, whereas markers D12S347 and D12S1703 defined a region of approximately 18 cM. Positional candidate genes are being screened for deafnesscausing mutations

Clinical and cytogenetic studies in intersex ewes

Nine Sarda x Lacaune ewes with intersexual characteristics and an infertility condition at the reproductive anamnesis were analysed. In order to make a diagnosis, we have evaluated their behaviour and performed clinical and laparoscopic examination of the reproductive tract, as well as cytogenetic analysis. The ewes showed basically a female phenotype but a clinical examination revealed a different degree of masculinization in the morphology of external genital organs. A shorter vagina was observed in female-like ewes and a hypertrophic clitoris in male-like ewes. Laparoscopic analysis evidenced the presence of testis in seven individuals and, for two of them, the gonadal position was subcutaneous. Different male characteristics in the nine subjects, were also observed in their behaviour with a different degree of masculinization. Their blood samples were used for determining the percentage of male cells on lymphocytes chromosome spreads by using the C-banding technique. The haematopoietic chimeras (XX/ XY) found in the lymphocytes confirmed the diagnosis of freemartinism for seven out of the nine subjects

Graft-<i>versus</i>-leukemia effects after allogeneic bone marrow transplantation are active also in the presence of clones with chromosomal anomalies in addition to the Ph chromosome

Two male patients with Philadelphia-chromosome (Ph+) chronic myelogenous leukemia (CML) underwent allogeneic bone marrow transplantation (ABMT) in the first chronic phase after busulfan treatment. In both cases, the donor was a sister, and engrafting was demonstrated by chromosome analyses which showed only donor cells in the BM. Cytogenetic relapse occurred 29 and 30 months after ABMT, respectively, when host cells reappeared: in both cases, the Ph and additional anomalies typical of the blastic phase of CML were evident. We then monitored the chromosome picture for 52 and 39 months, respectively: no striking evolution occurred, and cells with the Ph and additional anomalies persisted together with donor cells, which were a minority in the first patient and a great majority in the second throughout the observation period. A clinical relapse was observed in the first patient, but the disease never progressed to a blastic phase, whereas the second patient has not relapsed 7 years after ABMT. We reviewed data from the literature on cytogenetic relapse after ABMT in CML without clinical relapse, especially the 12 patients in whom cytogenetic relapse included chromosome anomalies in addition to the Ph, as in our patients. We suggest that graft-versus-leukemia (GVL) reactions in such patients are able to arrest progression of the leukemic blastic clone and prevent a possible relapse in blastic phase

Identification of a mutant allele of the androgen receptor gene in a family with androgen insensitivity syndrome: detection of carriers and prenatal diagnosis

We report the results of a molecular study of a large family segregating the complete form of the Androgen Insensitivity Syndrome (CAIS) in several family members from three generations. We identified the mutant allele by polymerase chain reaction (PCR) amplification of the short tandem repeat (CAG)n, highly polymorphic in the population, present in the first exon of the androgen receptor (AR) gene. In this family four different alleles were detected and one of these showed a perfect segregation with the disease. This study enabled us to identify the heterozygous females in this family. We think that this simple, indirect test, is also suitable for prenatal diagnosis of Morris' syndrome when the mother is heterozygous for the size of the short tandem repeat and one affected subject in the family may be studied

Pure partial trisomy of 6p12.1-22.1 secondary to a familial 12/6 insertion in two malformed babies

We describe two malformed infants with trisomy 6p12.1âp22.1 due to 12/6 interchromosomal insertion. The phenotypic data observed in these patients are compared chiefly with a case cytogenetically similar described by Villa et al. [A. Villa, E.G. Gomez, L. Rodriguez, R.H. Rastrollo, M.E. Martinez Tallo, M.L. Martinez-Frias, Interstitial tandem duplication of 6p: a case with partial trisomy (6)(p12p21.3), Am. J. Med. Genet. 90 (2000) 369â375]. All three infants are trisomic for a genomic segment which largely overlaps that reported as duplicated in previous cases, but with the addition of a more proximal segment, extending from 6p12 to 6p21. We suggest that some of their phenotypic anomalies are due to the trisomy of this chromosomal region. We also speculate on the possible role played by the TFAP2B (Transcription Factor AP2-beta) gene, which is one of the genes mapped on the duplicated segment

Partial trisomy of the long arm of chromosome 1: prenatal diagnosis, clinical evaluation and cytogenetic findings: case report and review of the literature

Partial trisomy of the long arm of chromosome 1 is a relatively rare cytogenetic anomaly. Its phenotype has still not been completely defined, because of the cytogenetic heterogeneity of the cases so far described. We report a prenatal case of partial 1q trisomy associated with partial monosomy 4q, secondary to balanced maternal translocation t(1;4). The trisomic segment extended from 1q31.1 to qter and the monosomy 4q was from 4q35.2 to qter. The phenotypic anomalies found by post-mortem and autopsy examinations were compared with those of similar cases reported in the literature. We performed standard cytogenetics and fluorescence in situ hybridization. Cerebral ventriculomegaly, present in our case, seemed to be a constant feature in partial 1q trisomies, so this cerebral malformation could be considered as the main echographic marker for this chromosomal imbalance and trisomy 1q should be added to the list of chromosomal abnormalities associated with ventriculomegaly