Evaluation of TUBB8 gene alterations in infertile women with oocyte maturation and cleavage arrest referred to Royan Institute.

RESEARCH QUESTION: Are TUBB8 gene variations present in Iranian infertile women with oocyte maturation arrest or embryo cleavage arrest? DESIGN: TUBB8 gene variations were investigated by polymerase chain reaction sequencing on blood samples from 16 women with oocyte maturation arrest and 12 women with cleavage arrest, collectively referred to as the experimental cohort, as well as 56 fertile women as the control group. The Exome Sequencing Project and dbSNP databases and the Genome Aggregation Database were used to search the frequency of corresponding variants. PolyPhen and SIFT were used to conduct in-silico analysis of gene variations and Align-GVGD was used to predict the effect of missense variants on proteins. The homology modelling and structure evaluation of variations was also checked. RESULTS: Two likely pathogenic variants [c.713C>T (p.Thr238Met), c.1054G>T (p.Ala352Ser)] were identified in patients with oocyte maturation arrest and one likely pathogenic variant [c.G763A, (p.Val255Met)] was identified in a patient with cleavage arrest. These changes were absent in controls. CONCLUSIONS: Three deleterious variants in TUBB8 related to oocyte maturation arrest or cleavage arrest and infertility were identified. TUBB8 variant screening for patients with oocyte maturation and cleavage arrest is recommended.

A Rare Case of Klinefelter Syndrome Patient with Quintuple Mosaic Karyotype, Diagnosed by GTG-Banding and FISH.

Klinefelter syndrome (KS) is the most common sex chromosomal disorder in men. Most of these patients show the 47,XXY karyotype, whereas approximately 15% of them are mosaics with variable phenotype. A 39-year-old male investigated for primary infertility, was clinically normal with small firm testes and elevated levels of FSH, LH and low level of testosterone. Total azoospermia was confirmed on semen analysis. Testicular histopathology revealed no spermatogenesis and absence of germ cells. Karyotype from whole blood culture showed cells with 47,XXY/46,XX/ 45,X/48,XXXY/ 46,XY mosaicism. The predominant cell line was 47,XXY (83.67%). This was confirmed by fluorescence in situ hybridization (FISH). Also the presence of a small population of cells with the 48,XXXY and 45,X karyotypes was detected by FISH. This case illustrates the utility of FISH as an adjunct to conventional cytogenetics in assess the chromosome copy number in each cell line of a mosaic.

Delineating the association between isodicentric chromosome Y and infertility: a retrospective study.

OBJECTIVE: To report on 14 infertile patients who had a de novo form of the same isodicentric (idic)(Yq) karyotype with variable degrees of mosaicism. DESIGN: Retrospective study and review of the literature. SETTING: Medical genetics laboratory in a research institute for reproductive biomedicine. PATIENT(S): Fourteen infertile patients, including 13 male patients and 1 female patient who had infertility with the same idic(Y) karyotype. INTERVENTION(S): Conventional cytogenetic methods, fluorescence in situ hybridization (FISH) on seminal germ cells and blood, and polymerase chain reaction (PCR)-based molecular approaches. MAIN OUTCOME MEASURE(S): Karyotype, FISH, and PCR results. RESULT(S): Cytogenetic results revealed abnormal Y chromosome: 45,X/46,X,idic(Y)(q11.22). The FISH technique on blood lymphocytes confirmed a rearranged Y chromosome, with two centromeres and two SRY signals, and marker chromosome with various levels of mosaicism. Moreover, aneuploidy of sex chromosomes was also detected in haploid seminal germ cells. Multiplex PCR analysis of blood samples demonstrated microdeletion in AZFb and AZFc loci. CONCLUSION(S): Because of the resemblance between inversion of chromosome Y and idics(Y), use of confirmatory techniques (e.g., FISH or PCR-based methods) could help prevent medical errors in healthcare systems and precisely delineate chromosomal aberrations in infertile patients when clinical data fail to clarify the cause of infertility.

Buthionine sulfoximine inhibits cytopathic effects and apoptosis induced by infection with AIK-HDC strain of measles virus.

BACKGROUND: Measles virus (MV) is a highly contagious agent which causes a major health problem in developing countries. We studied the effect of buthionine sulfoximine (BSO) on the replication of an AIK-HDC strain of MV and its induced apoptosis in Vero cell lines. METHODS: In this study, toxicity of BSO on Vero cells was investigated first, resulted in determination of sub-lethal or non-toxic concentration zone of BSO for cells. Next, anti-viral effect of BSO at various time limits was evaluated and virus titer was determined at each stage either as 50% tissue culture infective dose (TCID) 50 or by plaque assay method. Using specific anti-measles IgG, anti-viral effect of BSO on MV replication cycle was evaluated through indirect immunofluorescence assay, meanwhile presence of viral RNA was investigated by RT-PCR and gel electrophoresis. RESULTS: According to the experiments, BSO, at concentration of 50 microM, markedly inhibited the cytopathic effect (CPE) induced by MV. BSO also significantly inhibited apoptosis induced by MV. BSO either influences replication of MV genome, or may inhibit virion formation. CONCLUSION: These results suggest that the inhibition of CPE and apoptosis by BSO induced by MV may be associated with the effect of BSO on viral RNA genome. Therefore, it is suggested that MV infections can induce apoptosis through the activation of a common pathway that can be inhibited by BSO.