Complex cytogenetic and molecular-genetic analysis of males with spermatogenesis failure.

The chromosomal anomalies, microdeletions of AZF region of Y-chromosome and CFTR gene mutations have been studied among 80 infertile men with idiopathic spermatogenetic failure: 36 (45%) patients with aspermia, 19 (24%) patients with azoospermia and 25 (31%) patients with severe oligoasthenoteratozoospermia. In total 30% males with spermatogenetic failure genetic factor of infertility was observed. Karyotype anomalies were observed in 17.5% of infertile men, within 16.2% numerical and structural gonosomal anomalies and in 1.3%--Robertsonian translocation were revealed. In 11% males with spermatogenetic failure, Y-chromosome AZF region microdeletions were detected. The frequency of CFTR major mutation F508del among infertile men was 6.25%. 5T allele of polymorphic locus IVS8polyT was detected in 7.5% of examined men. The results obtained indicate the high complexity of cytogenetic and molecular-genetic studies of male infertility.

[Androgen receptor CAG gene polymorphism in men with azoospermia and oligozoospermia in Ukraine].

The number of CAG repeats of exon 1 of AR gene was determined in a group of 228 infertile males with azoospermia (n = 68) and oligozoospermia (n = 160) as well as in control group (124 proven fathers) by fluorescent polymerase chain reaction amplification followed by fragment analysis on automated fluorescent analyzer "A.L.F-express". The frequency of alleles with GAG-repeats < or = 18 was significantly higher (P < 0,01) in the group of patients with azoospermia (17,7%) comparing with the control group (2,4%) as well as in the group of patients with oligozoospermia (12,5%) comparing with the control group (2,4%). The frequency of alleles with CAG-repeats > or = 28 significantly differed (P < 0,01) between the group of patients with oligozoospermia (12,5%) and the control group (2,4%). Our data suggest an association between CAG repeats number and impaired spermatogenesis in azoospermic and oligozoospermic males.

[Y-chromosome microdeletions as a prognostic marker of male infertility].

The results of molecular-genetic study of Y-chromosome microdeletions in men with spermatogenesis failure and in patients with cryptorchism are presented. The molecular-genetic studies of regions AZFa, AZFb, AZFc in STS loci - sY84, sY86, sY127, sY134, sY254, sY255 and SRY gene have been performed. Y-chromosome microdeletions were detected in 13,3% infertile men with spermogram failure. The frequency of genetic (cyto- and molecular) abnormalities among boys with isolated cryptorchism was 4%. The results show the necessity of additional study ofgenetic factors ofcryptorchism development.

Monozygotic twins with congenital acute lymphoblastic leukemia (ALL) and t(4;11)(q21;q23).

Congenital acute lymphoblastic leukemia (CALL) is a rare disorder and is frequently associated with t(4;11)(q21;q23). To our knowledge this is the first case report of monozygous twins with CALL and t(4;11)(q21;q23).

Prognostic characteristics of surgical stage I endometrial adenocarcinoma.

PURPOSE: To evaluate and correlate the expression of pathologic characteristics, flow cytometric DNA content analysis, and estrogen and progesterone receptor levels with survival in patients with surgical Stage I endometrial carcinoma. METHODS AND MATERIALS: Hospital tumor registry records were surveyed, and this identified 232 patients diagnosed with endometrial adenocarcinoma between July 1, 1989, and December 30, 1993. DNA content analysis was performed on either paraffin-embedded or fresh tissue samples. Survival was calculated from the date of diagnosis by the Kaplan-Meier method. Postoperative irradiation (whole pelvis external beam therapy and low dose rate vaginal cuff brachytherapy) was delivered to patients felt to be at high risk for failure. RESULTS: One hundred seventy-one patients had Stage I tumors and were available for analysis. Patients with Stage 1C tumors had a statistically significant lower survival rate compared to patients with Stages IA or IB (p = 0.03 and p < 0.01, respectively). Patients with DNA content diploid tumors had a slightly increased (but nonsignificantly so) survival compared to patients with non-DNA content diploid tumors (p = 0.12). Logistic regression analysis failed to identify an independent prognostic factor that could predict for disease specific survival in patients with Stage I cancers. CONCLUSION: Logistic regression analysis did not identify a single independent prognostic factor in patients with Stage I tumors. Pathologic characteristics reported to predict survival advantage correlated with pathologic stage. Additional translational research is needed to identify molecular characteristics of tumors that may indicate more aggressive treatment for patients at high risk for recurrence.

Clinicopathologic correlation of DNA flow cytometric content analysis (DFCA), surgical staging, and estrogen/progesterone receptor status in endometrial adenocarcinoma.

DNA flow cytometric content analysis (DFCA) and estrogen (ER) and progesterone (PR) receptor levels are reported to be prognostic with regard to the malignant potential of endometrial adenocarcinoma. We retrospectively reviewed the records of 50 patients presenting with endometrial adenocarcinoma between July 1990 and December 1992, to determine the extent of any pathologic features reported at the time of hysterectomy. Patients whose tumors were nondiploid (aneuploid) by flow cytometry generally presented with a higher pathologic stage, higher grade, and more frequent lymph node involvement. In addition, the majority of clear cell and uterine papillary serous (UPS) adenocarcinoma were also nondiploid. Fourteen of 21 ER-positive tumors aneuploid, as were 18 of 37 PR-positive tumors. We also found DNA-A (DNA content aneuploid) patterns frequently associated with tumor characteristics implicated by other authors as related to aggressiveness. Further studies comparing the molecular biology of tumors to their clinicopathologic features and behavior are needed to fully understand the ultimate malignant potential.

Flow cytometric DNA content analysis of paraffin-embedded tissue derived from cervical carcinoma.

PURPOSE: Flow cytometric deoxyribonucleic acid (DNA) content analysis has been shown to be of prognostic importance in some cancers. There have been recent reports of a prognostic importance for DNA content analysis in cervical carcinoma. METHODS AND MATERIALS: We retrospectively reviewed the hospital and radiation oncology records of cervical carcinoma patients who presented between 1984-1990. RESULTS: A total of 101 archival paraffin-embedded blocks were processed, of which 77 were of technical quality for analysis. Thirty-five percent were found to be DNA content aneuploid (DNA-A) and 65% DNA content diploid (DNA-D). No statistical difference was found between the two groups in age at diagnosis, % S-phase, coefficient of variation (CV), or proliferative index (PI). A statistical difference was noted in the G2M phase between the two groups (p = 0.004). The median % S-phase was 8.4% in the DNA-D group. A statistical difference (p = 0.017) in survival was noted between the low and high % S-phase DNA-D groups. In patients who received radiation alone, high-PI patients had improved survival compared to low-PI patients. No statistical difference in survival was noted in the high % S-phase DNA-D group and DNA-A group (p = 0.28). Proportional Hazard (Cox) Regression found clinical stage the only independent prognostic indicator for survival. CONCLUSION: Flow cytometric DNA content analysis is being used more frequently in the management of different malignant tumors. Our study shows that DNA content analysis is useful in determining the prognosis and survival outcomes in cervical carcinomas and may aid in predicting outcome to certain types of treatment regimens.

Myelodysplastic syndrome after cisplatin therapy.

Therapy-related myelodysplastic syndrome (tMDS) and acute nonlymphocytic leukemia (tANLL) are known late complications of cytotoxic drug therapy for hematologic malignancies, solid tumors, and nonmalignant conditions. The alkylating agents are often the causative agents, but a few reports have implicated cisplatin as an etiologic agent. Cisplatin has a significant impact on the treatment of a number of malignant neoplasms, including testicular and ovarian cancer, and is a part of several clinical trials for squamous cell carcinoma of the head and neck region. Given its increasing use, a complication as significant as tMDS is potentially important. In this article, the authors describe the case of a patient who had myelodysplastic syndrome develop after successful treatment for laryngeal cancer with cisplatin. The treatment included cisplatin in combination with 5-fluorouracil, followed by radiation therapy. The authors also present a review of articles in the literature regarding tMDS and tANLL occurrence after treatment with cisplatin-containing regimens. The authors conclude that cisplatin can be a leukemogenic agent. The drug may potentiate the leukemogenic effects of other alkylating agents and drugs that inhibit topoisomerase II action.

Cytogenetic studies of carcinoma in situ of the bladder: prognostic implications.

Carcinoma in situ of the bladder has traditionally been considered a lethal disease by most urologists and as such it has been treated most frequently in an aggressive manner. Recent investigations have suggested that carcinoma in situ of the bladder may, in fact, be a complex of diseases that exists in at least 2 distinct disease forms, 1 aggressive and 1 relatively nonaggressive. We studied the cytogenetics of 17 patients diagnosed clinically and pathologically to have carcinoma in situ, and found a positive correlation of karyotype complexity (numerical and structural changes) with disease course. Superficial tumors with normal karyotypes remained superficial throughout the study and continued to exhibit a nonaggressive course, while patients whose tumors had an abnormal karyotype at diagnosis expressed a much more aggressive course that ultimately developed into invasive disease. Nonrandom chromosomal changes involving chromosomes 1, 5, 8 and 11 were observed in these tumors. Therefore, cytogenetic evaluation may prove to be an important guide in helping to determine an appropriate treatment course for patients with carcinoma in situ of the bladder.

Characterization of a new human multiple myeloma cell line, UMJF-2, which suppresses antibody production by B-lymphocytes in vitro.

A new human plasma cell line, UMJF-2, has been derived from the bone marrow of a patient with multiple myeloma. Morphological studies disclosed large nucleoli, moderate numbers of mitochondria, and scant endoplasmic reticulum consistent with a plasmablastic morphology. The cells have immunologic characteristics of early plasma cells, including intense expression of cytoplasmic IgG-lambda and weaker, but discernible, expression of surface IgG-lambda. Cell surface antigens defined by the monoclonal antibodies OKT10 (CD38) and PCA-1, characteristic of mature plasma cells, and B1 (CD20), B4 (CD19), and I-2 (HLA-DR), characteristic of earlier stages of B-lymphocyte differentiation, are present on UMJF-2 cells. Cytogenetic studies reveal the presence of trisomy 12. UMJF-2 does not contain the Epstein-Barr virus by Southern blot analysis. Tissue culture media conditioned by these cells contains a soluble immunosuppressive factor, capable of inhibiting pokeweed mitogen induced IgM secretion by normal human B-lymphocytes. UMJF-2 provides a model for the study of the pathogenesis of polyclonal hypogammaglobulinemia in human multiple myeloma.

Apparent correlation of sex chromosome loss and disease course in urothelial cancer.

In recent years, interest in the genetics of various solid tumours has increased dramatically. Over the last several years, our laboratory has pursued genetic studies of transitional cell carcinoma of the bladder. We have studied the cytogenetics and disease course of more than 100 patients, achieving successful cultures in more than 75%. In these patients, we observed the loss of a sex chromosome (either X or Y) with a frequency of 40%. We stratified patients with loss of sex chromosomes into three groups: 1) presence of markers or chromosome rearrangements, 2) aneuploidy without marker chromosomes, and 3) loss of a sex chromosome only. The relevancy of both the natural history and potential impact of sex chromosome loss to the disease course, including invasive potential, will be discussed in detail for each of the three groups.

X chromosome instability associated with familial Turner syndrome.

A family with two members (two generations) exhibiting Turner syndrome is described. Cytogenetic studies on these individuals showed the presence of multiple X chromosome changes. Evidence is presented to show that the maternally inherited X chromosome is the chromosome involved in the structural alterations observed. The effect of a tendency of the maternal X chromosome to break at specific sites on the development of the Turner phenotype and abnormal karyology is discussed.

Atypical gonadal dysgenesis: the role of Yq in determining phenotype and malignancy risk.

Two cases of atypical gonadal dysgenesis are described. Phenotypes are attributed to unique karyotypes derived from abnormal zygotic states modified by secondary cell line loss. A model is presented which proposes and defines a regulatory locus of Yq (Y-HT) and comments are made regarding the risk of malignancy in such patients.

Aarskog syndrome: full male and female expression associated with an X-autosome translocation.

We describe a male infant with the Aarskog syndrome and his equally prominently affected mother. The propositus and his mother have a balanced X-autosome translocation which originated in her. We postulate that the mother's translocation resulted in a presumed de novo point mutation for the Aarskog locus and that she has nonrandom inactivation of her structurally normal X. The full expression of the syndrome in the mother is compared to the partial expression in reported females. It is concluded that the Aarskog syndrome is an X-linked disorder and that the locus for the syndrome is at Xq13.

Karyotype evolution in a transformed rat cerebral endothelial cell line.

Primary cultures of rat microvascular endothelial cells were transformed, in vitro, by exposure to Rous sarcoma virus. Transformed cells were followed and evaluated cytogenetically through numerous passages. Highly specific karyotypic changes in karyotype (both structural and numerical) were documented. These changes became established and intimately involved in further "karyotypic evolution". The findings were reproducible, and when considered in the light of the literature suggest regular patterns of karyotypic change in rat tumors. The in vitro methodology utilized promises to be of practical value in the study of the early stages of malignancy.

Double aneuploidy: partial trisomy 21 and XO/XXX in a family with 12/21 translocation.

A female patient with mild mental retardation with spatial perceptual difficulties, microcephaly, depressed nasal root, receding chin, webbed neck, low hairline, shield chest, cubitus valgus, scoliosis and dermatoglyphic findings not characteristic of Down's syndrome is reported. In addition to X/XXX, she had a partial trisomy 21 of the short arm-centromere-proximal long arm segment due to maternal t(12;21) translocation. Two phenotypically normal siblings carried the balanced translocation.