Karyotype In Peripheral Lymphocytes Research Articles

Yolk sac tumor and dysgerminoma in the left gonad following gonadoblastoma in the right gonad in a 46,XY DSD with a novel SRY missense mutation: a case report

BackgroundApproximately 10–15% of 46,XY disorders of sex development (DSDs) have an SRY mutation residing in the high mobility group (HMG) domain. Here, we present a case of 46,XY DSD caused by a novel missense mutation in the HMG region of SRY rapidly progressing to germ cell tumors (GCTs).Case presentationAn adolescent female (15 years old) exhibiting primary amenorrhea was later diagnosed as a 46,XY female with bilateral gonadal dysplasia on the basis of peripheral lymphocyte karyotype 46,XY and a novel missense mutation in SRY (c.281 T > G, p.L94R). The novel missense mutation (c.281 T > G, p.L94R) and its adjacent region were conserved. Protein structure analysis showed that the mutant site was located in the middle of the HMG domain, and the mutant protein had a diminished ability to bind to DNA. Imaging examination revealed an adolescent female with a naive uterus. Laparoscopy and initial pathological examination revealed left gonadal dysplasia and right gonadal dysplasia with gonadoblastoma (GB). Right gonadectomy by laparoscopy was performed upon consent from the patient’s parents. Less than 1 year postoperatively, the left gonadal gland deteriorated as observed by the findings of a mass in the left adnexal region by pelvic MRI and serum AFP > 1000 ng/ml by serological tests, and then total hysterectomy and adnexal and left gonadectomy by laparoscopy were performed. The GCT stage was classified as stage Ic according to FIGO. At this time, pathologic examination showed that the left gonad had progressed to yolk sac tumor and dysgerminoma. The patient underwent chemotherapy post-operatively but developed type III myelosuppression and tumor recurrence several months later.ConclusionsThe patient initially presented with right gonadoblastoma but chose only right gonadectomy by laparoscopy to preserve the female sex characteristics, which resulted in rapid deterioration of the left gonad and poor treatment outcomes. This case demonstrates the importance of early genetic diagnosis and treatment of 46,XY female DSD.

Clinical spectrum and Cytogenetic characterization of patients with Turner Syndrome – Twin case report

Diagnosis of Turner syndrome (TS) is usually made in mid-childhood, where 50% of the patients have 45, X karyotype in peripheral lymphocytes, which results from haploinsufficiency of the genes that commonly escape X-inactivation. 30%–40% have mosaicism of different forms, like 45,X/46,X,dic(Xp)/46,X,idic(Xq) and less commonly 45,X/46,XY. The goal of this case report is to analyze the cytogenetic and molecular characterization of two cases with dicentric X chromosomal abnormalities with varying degrees of mosaicism, demonstrating shared clinical features of TS. Combined conventional cytogenetic analysis, centromere banding, and fluorescence in situ hybridization (FISH) was done for the patients who presented with short stature and irregular menstrual cycles. Chromosome studies showed two cell lines: one with a single copy of X chromosome (45,X) and the other with a structural variation in X chromosome (isodicentric X chromosome), which is described as a Turner variant. C-banding also revealed the presence of two centromeres. Metaphase FISH with centromere probes for X revealed two mosaic cell lines: one with 45,X and a second one showing isodicentric X chromosome. The accurate diagnosis and characterization of a genomic imbalance in patients with sex chromosome disorders are essential for evaluating phenotype–karyotype correlations, genetic counseling, and having a clinical follow-up.

Clinical features and Y chromosomal abnormalities in patients with karyotype chimerism of 45, XO/46, XY

Objective To increase the knowledge of clinical features and Y chromosomal abnormalities in patients with mosaic karyotype 45,XO/46,XY.Methods The clinical features,peripheral lymphocyte karyotype,sex hormones,treatment,and follow-up information were retrospectively reviewed.Results (1) 7 patients with 45,XO/46,XY were included.Their social gender was 3 male and 4 female,and the age by diagnosis was (14 ± 3) years (range 11-20 years).(2) Patients presented with short stature (n =7) and other specific somatic signs similar to Turner's syndrome.(3) The external genitalia varied from complete male to complete female manifestations.There were normal penis (n =1),hypospadia (n =2),enlarged clitoris and combined opening with ureter and vagina (n =1),and wholesome vagina opening (n =3).By physical examination and ultrasound,testes were detected in 3 patients,1 patient had testes and ovary-like gonad,and 1 had ovary-like gonad,and no gonads were detected in 2 patients.(4) Among 7 patients,4 had obviously abnormal Y chromosome.1 patient had micro-deletion on Y chromosome by Multiplex Ligation Dependent Probe Amplification (MLPA).(5) 4 patients had therapy with (rhGH).Gonads were removed from P4,showing ovary-like tissue.3 patients received sex hormone replacement.Conclusion Patients with 45,XO/46,XY mosaic karyotype manifest short stature and other clinical features of Turner's syndrome.The presence of testis may result in different extent of virilization.The mosaic karyotype and failure of spermatogenesis may be caused by abnormal Y chromosome.Treatment includes rhGH therapy,removal of dysgenic gonads and hormonal replacement. Key words: 45, XO/46, XY ; Turner's syndrome ; Karyotype ; Y chromosome

A successful second delivery outcome using refrozen thawed testicular sperm from an infertile male true hermaphrodite with a 46, XX/46, XY karyotype: case report

We report a successful second delivery of a healthy infant fathered using refrozen thawed testicular sperm from an infertile male chimera. We also examined sex chromosome distribution of the seminiferous tubule. Intracytoplasmic sperm injection (ICSI) was performed using the remaining refrozen testicular sperm, which had been stored during the first treatment. Biopsied testicular cells were examined by fluorescence in situ hybridization (FISH) and the peripheral lymphocyte karyotype was tested using a G-band. Following ICSI, a second pregnancy was established, and a healthy girl was successfully delivered at 40 gestational weeks without complications. Although the husband's lymphocyte chromosomal analysis revealed a 46, XX [28]/46, XY [2] karyotype, the seminiferous tubule cells on histological examination by FISH were chimeric sex chromosome type XX [18]/XY [82]. In conclusion, this is a very rare case report of a successful subsequent delivery of a healthy infant (46, XX) from an infertile true hermaphrodite (46, XX/46, XY) using refrozen thawed testicular sperm. The seminiferous tubule cells' karyotype ratio differed from that of the lymphocytes.

Cleft palate and complex chromosome rearrangements

Two of three unrelated children with de novo congenital complex chromosome rearrangements (CCR) with more than four chromosome breaks had cleft lip and palate as one of several congenital anomalies. In patient 1, unilateral complete cleft of the primary and secondary palates accompanied severe ectrodactyly, bilateral posterior choanal atresia and several minor congenital anomalies. Karyotypes of peripheral lymphocytes and skin fibroblasts showed five derivative chromosomes with six break points. There were two translocations, t(2;5), t(3;11) and an interstitial deletion, del(13)(q12q14). Patient 2 had a bilateral complete cleft of the lip and palate, in addition to slow pre- and postnatal growth and minor congenital anomalies. Peripheral lymphocytes and palatal mucosa fibroblasts karyotypes showed five derivative chromosomes with six break points. A partial deletion of 10p, two translocations, t(2;3), t(7;18) and an inversion of the derivative chromosome 2 were present. In both patients, a "major catastrophe" of unknown etiology in one of the parental gametes appeared to be the event leading to the stable CCR without evidence of persistent chromosome instability. All four parents had normal karyotypes. The presence of palatal clefts in these patients indicates that dysmorphologists and pediatricians have to consider CCR whenever taking care of a patient with cleft palate, particularly if additional anomalies, no matter how subtle, are present. The detection and interpretation of the latter anomalies are essential for the diagnosis and management of these patients. Accurate cytogenetic diagnosis determines the short- and long-term prognosis and facilitates genetic counseling in regard to life-span, quality of life and reproductive plans of patients and parents.

Report of Fertility in a Woman with a Predominantly 46,XY Karyotype in a Family with Multiple Disorders of Sexual Development

We report herein a remarkable family in which the mother of a woman with 46,XY complete gonadal dysgenesis was found to have a 46,XY karyotype in peripheral lymphocytes, mosaicism in cultured skin fibroblasts (80% 46,XY and 20% 45,X) and a predominantly 46,XY karyotype in the ovary (93% 46,XY and 6% 45,X). A 46,XY mother who developed as a normal woman underwent spontaneous puberty, reached menarche, menstruated regularly, experienced two unassisted pregnancies, and gave birth to a 46,XY daughter with complete gonadal dysgenesis. Evaluation of the Y chromosome in the daughter and both parents revealed that the daughter inherited her Y chromosome from her father. Molecular analysis of the genes SOX9, SF1, DMRT1, DMRT3, TSPYL, BPESC1, DHH, WNT4, SRY, and DAX1 revealed normal male coding sequences in both the mother and daughter. An extensive family pedigree across four generations revealed multiple other family members with ambiguous genitalia and infertility in both phenotypic males and females, and the mode of inheritance of the phenotype was strongly suggestive of X-linkage. The range of phenotypes observed in this unique family suggests that there may be transmission of a mutation in a novel sex-determining gene or in a gene that predisposes to chromosomal mosaicism.

A 45,X/47,XYY/46,XY Karyotype and Y Chromosome Microdeletion in an Infertile Male

A 45,X/47,XYY/46,XY Karyotype and Y Chromosome Microdeletion in an Infertile MaleInfertility affects 10-15% of all couples and the male factor is responsible in approximately half of them. The advances in assisted reproduction techniques and genetic diagnoses have increased the percentage of couples who are able to conceive. We report on an infertile male patient with azoospermia, referred to our laboratory before intracytoplasmic sperm injection (ICSI), whose peripheral lymphocyte karyotype showed a 45, X/46, XY mosaicism. A Y chromosome (Yq) microdeletion was detected in the azoospermia factor (AZF) regions, AZFb and AZFc. The karyotype from testicular tissue cultures was mosaic 45, X[13]/47, XYY[12]/46, XY[25]. Fluorescence in situ hybridization (FISH) performed on blood lymphocytes and testicular cells showed a 47, XYY cell line in both, in 8 and 23% of cells, respectively. Histological examination of the testicular biopsy revealed very few seminiferous tubules, which were hyalinized and composed only of Sertoli cells. Intracytoplasmic sperm injection was not suggested to the patient due to the sex chromosome mosaicism, Yq microdeletion and biopsy findings.

A successful pregnancy outcome using frozen testicular sperm from a chimeric infertile male with a 46, XX/46, XY karyotype: Case report

We report a very rare case of the successful delivery of a healthy infant fathered by an infertile chimera. ICSI was performed using frozen sperm. The karyotypes of peripheral lymphocytes were examined with a G-banding stain. Chromosomal analysis prior to ICSI revealed a 46, XX [28]/46, XY [2] karyotype chimera. As an infant, the subject was diagnosed as a true hermaphrodite, and underwent a hysterectomy and oophorectomy. A small number of sperm were found in minced testicular tissue, and they were stored for ICSI. To the best of our knowledge, this is the first case report of a successful pregnancy and delivery of a healthy infant fathered by an infertile chimera (46, XX/46, XY) following ICSI using frozen testicular sperm.

Cytogenetic and molecular-cytogenetic studies of Rett syndrome (RTT): a retrospective analysis of a Russian cohort of RTT patients (the investigation of 57 girls and three boys)

Rett syndrome (RTT) is a severe neurodevelopmental disorder with an incidence of 2.5% in mentally retarded girls in Russia. We have performed cytogenetic studies of 60 patients (57 girls and three boys) with a clinical picture of RTT, selected according to the criteria for diagnosis of RTT defined by B. Hagberg et al. in 1996. Collection of DNA samples and fixed cell suspensions of RTT patients (37 girls and two boys) and their parents (27 patients) was established for molecular studies, for example analysis of MECP2 mutations in a Russian cohort of RTT patients. Among 60 patients 57 girls with a clinical picture of RTT had normal female karyotype (46,XX), one boy had normal male karyotype in peripheral lymphocytes (46,XY) and two boys had a mosaic form of Kleinfelter's syndrome (47,XXY/46,XY) in peripheral lymphocytes or muscle cells (with MeCP2 mutation R270X). Twenty-four mothers and parents of RTT girls had normal karyotype, two mothers had mosaic forms of Turner syndrome (45,X/46,XX) and one had mosaic karyotype (47,XX,+mar/48,XXX,+mar). We analyzed chromosome X in lymphocytes of 57 affected girls with a clinical picture of RTT using the 5-bromo-2′-deoxyuridine+Giemsa staining technique. A specific type of inactive chromosome X (so-called type ‘C’) with unusual staining of chromatin in the long arm of chromosome X was found in 55 (from 57) girls with RTT. This technique was positively used for presymptomatic diagnosis of RTT in five girls in earlier stages of the disease. We believe that the phenomenon of altered chromatin conformation in inactive chromosome X could be used as a laboratory test for preclinical diagnosis of the RTT.

Deletion (9) (p13.1 p21.1)

We report on a 22-month-old girl with minor facial anomalies, global developmental delay, growth retardation, seizures, and leukoencephalopathy. Initial clinical assessment suggested the diagnosis of Williams syndrome. Results of fluorescence in situ hybridization testing for elastin were normal. However, chromosome analysis showed a 46,XX,del(9)(p13.1p21.1) karyotype in peripheral lymphocytes. Parental chromosomes were normal, indicating a de novo deletion. This patient's manifestations are compared with those of two other cases with overlapping deletions of the proximal short arm of chromosome 9.

Molecular analysis of a patient with Beckwith-Wiedemann syndrome, rhabdomyosarcoma and renal cell carcinoma

We described a patient with Beckwith-Wiedemann syndrome associated with rhabdomyosarcoma (RMS), and renal cell carcinoma (RCC). Karyotypes of peripheral lymphocytes and RMS cells were normal. DNA analyses showed maternal loss of heterozygosity (LOH) at 11p15 region in RMS but not in RCC. The insulin-like growth factor II gene (IGF2) was found to be expressed at a moderate level in RMS but not in RCC by in situ hybridization. Each of parental allele-derived IGF2 transcript was detected in RCC, while maternal allele-derived transcript was weak in RMS because of maternal LOH. These results suggest that (1) loss of imprinting (LOI) of IGF2 might be responsible for BWS, (2) on the other hand, LOI itself might not induce tumor occurrence in tissues where the control of tissue-specific expression of IGF2 is maintained, (3) increased expression of IGF2 due to maternal loss of a putative controller gene for IGF2 at 11p15 might predispose to sustaining tumorigenic mutations and tumor progression, (4) loss of a putative onco-suppressor gene at 11p15 might induce RMS occurrence. The cause of RCC was thought to be different from that of RMS.

XX TRUE HERMAPHRODITISM ASSOCIATED TO MULTIPLE MALFORMATION. ABSENCE OF Y-SPECIFIC DNA SEQUENCES AND NORMAL H-Y ANTIGEN EXPRESSION

True hermaphroditism occurs when ovarian and testicular tissues are present in the same patient and usually is not associated to other malformation. We studied a patient with ambiguous genitalia from day 6 of life, phallus measured 3 cm, urethral and vaginal orifices were separate, slight posterior labial fusion and left(L) gonad palpable in the inguinal region, X̄ chromatin on buccal smear was positive 28% and karyotype in peripheral lymphocytes 46.XX. hCG stimulation resulted in great testosterone increase (<10 to 832 ng/dl). Ultrasonography revealed uterus and tubes but no intrabdominal gonads. The patient was assigned a female sex and at 16 month exploratory laparotomy revealed uterus, tubes, an ovary on the right(R) and an ovotestis on the 1, which was removed. The patient had multiple malformation; cranial assymetry, telecantum greater on the L, low nasal bridge, fat and cutaneous hypertrophy in lower limbs and gluteus and with keratosis, specially on the dorsum of R foot. On the feet: normal halux on L and with clinodactiTia on the R, 2nd. finger with bilateral macrodactilia and R Clinidactilia and 4th. finger with bilateral campodactilia and L macrodactilia. Growth was normal (Ht. 73.6 cm at ly. 1mo.). The parents were young and not consanguineous and denied affected relatives. The mother was medicated, with piperidolate hydrochloride in the first trimester and ampicilin and cephalotin for urinary infection on the 4th, month of gestation, The patient was born full term with 3.400 g and 46 cm lenqth and had hypoglycemia. The patient had no Y-specific UNA sequences, as detected by ZFY probe in Southern hybridizations (the probe corresponds to a Y chromosome region encompassing the socalled testis determining factor gene, or TD(-). Expression of H-Y antigen assayed with monoclonal anti-H-Y antibody and an ELISA was normal. Although hermaphroditism and malformations could be unrelated, one can speculate that alteration of a single autosomal or X-linked locus could have caused the patients's condition. This work was sponsored partially by a grant from

Chromosome aberrations in peripheral lymphocytes of male homosexuals

Karyotypes of peripheral lymphocytes of 19 male homosexuals showed increased hypodiploidy. Chromosomes #19 and #20 were most frequently lost. Also, structural chromosome aberrations frequently occurred consisting chiefly of translocations and simple chromosome breaks. Terminal deletions, inversions, and isochromosomes occurred less commonly. In three of the cases, 100% of the cells were involved in a pericentric inversion of a chromosome #9. Chromosomes #3 in p21.1 and 1 in p32.3 were repeatedly affected. Structural aberrations were seen less frequently in men with acquired immunodeficiency syndrome(AIDS) and AIDS-related complex than in asymptomatic homosexuals. The hypodiploidy with preferential loss of chromosomes was constantly present. The marker chromosomes and simple breaks at repeated sites are another manifestation of damage to the immune system in these male homosexuals from Greenwich Village in New York City. The chromosomal damage was potentially the result of exposure to amyl and butyl nitrites, viral infections, or immunologic reactions to sperm, which crossreact with lymphocytes.

An Autopsy Case of Primary Acquired Sideroblastic Anemia with Isochromosome 17 Aberration

An autopsy case of primary acquired sideroblastic anemia (PASA) is described. Cytogenetic studies showed an isochromosome 17 (17qi) not only in diploid but also in octaploid cells from the bone marrow, whereas the karyo-type of peripheral lymphocytes was normal. Postmortem examinations revealed hyperplasia of the three myeloid cell lines in the bone marrow and extramedullary hematopoiesis in the liver and spleen, but no evidence of leukemia. The present case provides cytogenetic evidence that PASA is a disorder derived from an uncommitted stem cell mutation and is clonal in nature.