Chromosome fragility in cattle with chronic enzootic haematuria

Chronic enzootic haematuria (CEH) is a severe syndrome due to prolonged ingestion of toxic principles of bracken fern, such as quercetin and ptaquiloside. Little information is available on chromosomal instability of cattle with access to bracken fern and suffering from CEH. In the present study, 45 cattle, aged from 7 to 12 years and pastured in the south of Italy, were cytogenetically investigated for the first time in search of both chromosomal aberrations (aneuploidy, gaps, chromatid breaks, chromosome breaks and fragments) and sister chromatid exchanges (SCEs). Of these animals, 30 (group 1) had access to bracken fern and showed signs of CEH, and 15 (group 2; control) did not. Percentage of abnormal cells (aneuploidy, chromatid breaks, chromosome breaks and fragments) was higher in animals affected by CEH (34.7%, group 1) than that (24.3%) reached in the control (group 2). The same results were achieved when including gaps. Indeed, the mean number of cells with structural aberrations excluding gaps (chromatid breaks, chromosome breaks and fragments) per cell was higher (P<0.001) in animals affected by CEH (0.16+/-0.36) than that (0.09+/-0.29) found in the control. Chromosome fragility in cells of animals affected by CEH was also confirmed when applying the SCE test: statistically higher levels (P<0.001) of SCEs were observed in animals with CEH (7.35+/-3.59 SCE/cell, group 1) than those in the control (5.40+/-2.68 SCE/cell).     

Sialic acid and GM3 ganglioside expression in papillomavirus-associated urinary bladder tumours of cattle with chronic enzootic haematuria

This study was based on 30 papillomavirus-associated urinary bladder tumours from cattle with chronic haematuria, the animals having been kept since birth on pasture rich in bracken fern. The ganglioside content was assessed and compared with that of normal bovine urinary bladders, which was shown to be 28.6+/-3.3 (mean+/-SD) microg of lipid-bound sialic acid per gram of fresh tissue. In neoplastic bladder samples this value was higher but variable (120.9+/-80.6 in benign tumours, and 94.7+/-45.7 in malignant tumours). The main ganglioside, GM3, represented ca 75% of the total ganglioside mixture in normal tissues and 50-80% in tumour samples. GM1, GM2, GD1a, GD3 and FucGM1 were found as minor components. The study suggested that GM3 ganglioside may have a crucial role in "downregulation" of the metastatic potential of bovine urothelial cancers.     

Urinary bladder lesions in bovine enzootic haematuria

In cattle, bracken fern chronic toxicity is characterized by the presence of multiple tumours in the bladder (bovine enzootic haematuria). From October 1999 to March 2003, 433 urinary bladders with macroscopical lesions were collected in the slaughterhouse of S?o Miguel Island (Azores, Portugal), an endemic area where Pteridium aquilinum infestation in pastures is high. Bladder lesions were divided into three main categories (inflammatory lesions, non-neoplastic epithelial abnormalities and tumours) and described in detail. In some cases, neoplastic growth was confined to a single site, but in most cases multiple tumours developed within the same bladder. Epithelial tumours alone were present in 51.2% of the affected bladders, mesenchymal tumours alone in 17.4%, and both epithelial and mesenchymal tumours in the remaining 31.4%. The large number of tumours examined (870) revealed new categories not yet included in other veterinary classification systems, namely, inverted papilloma, papillary neoplasm of apparent low malignant potential, and haemangioendothelioma.     

Semi-nested PCR for detection and typing of bovine Papillomavirus type 2 in urinary bladder and whole blood from cattle with enzootic haematuria

Bovine Papillomavirus type 2 (BPV-2) and chronic intoxication by bracken fern ingestion were associated with urinary bladder lesions and the clinical signs of enzootic haematuria in adult cattle. Clinically enzootic haematuria is characterized by intermittent haematuria followed by animal death. Enzootic haematuria causes considerable economical impact on extensive cattle breeding worldwide. The demonstration of BPV-2 participation in the etiology of bovine urinary bladder carcinoma by conventional virological methods is not easy and the integrity of epidemiological studies relies on methods that are sensitive and specific for BPV-2 detection and typing. A multiplex-PCR was evaluated for BPV-2 L1 gene and bovine mitochondrial genome ND5 gene (internal control) detection followed by a second round of BPV-2 amplification by a semi-nested PCR (SN-PCR). Six skin papilloma samples were used for PCR technique development. Twenty-two urinary bladder samples from symptomatic (n = 12) and asymptomatic (n = 10, control group) cows and 25 blood samples from cows grazed on enzootic haematuria-endemic (n = 14) and enzootic haematuria-free (n = 11, control group) geographical regions of Parana State, Brazil were analyzed. The SN-PCR detected BPV-2 in seven urinary bladder and 10 whole blood samples collected from cows with enzootic haematuria and in one urinary bladder and one whole blood samples of asymptomatic cows. The specificity of the amplicon was performed by restriction fragment length polymorphism and sequence analysis. The SN-PCR technique developed in this study will make possible the realization of diagnosis and comparative epidemiological studies to evaluate BPV-2 infection rates in cattle, and the association of this infection with bracken fern chronic intoxication in the etiology of enzootic haematuria and opens the possibility of ante mortem studies by lymphocytes analysis.     

Chromosome 1 aberrations in cancer

Evidence for chromosome #1 involvement in structural rearrangements in cancer is reviewed. There have been adequate studies of cancer at most of the common sites, and at all of these, nonrandom chromosome #1 involvement has been demonstrated. In general, a variety of changes is encountered, irrespective of the site; most commonly, however, the changes result in the duplication of long arm material. It seems that these nonrandom changes, which tend to occur at a relatively late stage, may contribute to the progression of all forms of cancer. However, a small number of chromosome #1 aberrations are also now known, which may represent specific and possibly initiating changes in particular forms of cancer. These include short arm deletions in neuroblastoma and translocations in leukemias and myelodysplasia.     

Chromosome aberrations in nine patients with ovarian cancer

Cytogenetic analysis was performed on 13 tumor specimens (six solid tissues and seven effusions) from nine patients with various types of ovarian cancer. Eight of these patients had not received cytotoxic therapy prior to the initial karyologic assessment. Extensive and complex numerical and structural alterations were seen in nearly all specimens. Consistent (clonal) abnormalities were found in each case, but karyotypic heterogeneity within a tumor was a consistent finding in this series. Aberrations of chromosomes 1, 3, 6, 7, 10, and 12 were each observed in five or more patients. Although no specific recurring translocations were observed, partial deletions of 3p, 6q, 8p, and 10q were each seen in three different cases. Breakpoints tended to recur at several chromosomal band regions, some of which appear to be near the known loci of certain protooncogenes. Double minute chromosomes were seen in one patient, and a homogeneously staining region was found in another. Karyotypic analysis was performed on one patient both before and after initiating chemotherapy, and the chromosome pattern became more complex after treatment. Overall, our findings indicate that karyotypes in newly diagnosed, untreated patients with ovarian cancer can be extremely complicated, and that the extent of chromosome change may increase with tumor progression. Furthermore, the recurrence of specific regional chromosome losses suggests that these sites contain genes whose loss plays a role in the formation of some ovarian tumors.     

Chromosome aberrations in B-cell chronic lymphocytic leukemia. Pathogenetic and clinical implications

Chromosome analyses were performed on leukemic cells from 102 patients with B-CLL, of whom 84 were untreated. B-cell mitogen-induced CLL cells yielded suitable metaphases in 85 patients, and 55 showed clonal chromosomal aberrations. Trisomy 12 was found in 26 patients. In nine patients the + 12 was a single aberration. A 14q + chromosome or deletions of the long arm of chromosomes 6, 11, or 13 were other recurrent aberrations. Patients with Rai stage I or more had more frequently clonal aberrations than patients with stage 0 disease (p less than .02). Patients with clonal aberrations had poorer 5-year survival than those with a normal karyotype (p less than .05). Patients with a high percentage of abnormal metaphases in the sample had poorer prognosis than patients with high admixture of normal metaphases (p less than .01). Of the specific clonal aberrations those with 14q + or trisomy 12 tended to have slightly poorer and those with 6q- or structural aberrations involving the long arm of chromosome 13 tended to have better prognosis than patients with other chromosomal aberrations. A complex karyotype tended to be an adverse prognostic sign. Clonal evolution is rare: complex karyotypes are found at diagnosis and clones with single aberrations did not acquire additional chromosome aberrations despite progressive disease and treatment. Nine hundred and seventy-nine published cases are reviewed, and pathogenetic mechanisms, such as oncogenes and gene dosage, are discussed.     

Chromosome aberrations in B-cell chronic lymphocytic leukemia: reassessment based on molecular cytogenetic analysis

In B-cell chronic lymphocytic leukemia (B-CLL) clonal chromosome aberrations are detected in approximately 40–50% of tumors when using conventional chromosome banding analysis. Most studies find trisomy 12 to be the most frequent chromosome aberration, followed by structural aberrations of the long arm of chromosomes 13 and 14. Trisomy 12 and the ”14q+” marker are associated with shorter survival times, while the patients with 13q abnormalities have a favorable outcome, similar to those with a normal karyotype. The development of molecular cytogenetic techniques has greatly improved our ability to detect chromosome aberrations in tumor cells. Using fluorescence in situ hybridization, chromosome aberrations can be detected not only in dividing cells but also in interphase nuclei, an approach referred to as interphase cytogenetics. The prevalence of specific aberrations in B-CLL is currently being reassessed by interphase cytogenetics. By far the most frequent abnormality are deletions involving chromosome band 13q14, followed by deletions of the genomic region 11q22.3-q23.1, trisomy 12, deletions of 6q21-q23, and deletions/mutations of the TP53 tumor suppressor gene at 17p13. The evaluation of the true incidence of these aberrations now provides the basis for more accurate correlations with clinical characteristics and outcome. Deletions/mutations of the TP53 gene have been shown to be associated with resistance to treatment and to be an independent marker for poor survival. 11q deletions have been associated with extensive nodal involvement, rapid disease progression, and short survival times. Whether trisomy 12, 13q14, and 6q deletions have a prognostic impact awaits further study. The application of these molecular cytogenetic techniques will also contribute to the identification of the pathogenetically relevant genes that are affected by the chromosome aberrations in B-CLL. Received: 2 February 1998 / Accepted: 31 March 1998     

Chromosome aberrations in an alpha-fetoprotein-producing hepatoblastoma.

The cytogenetic findings of an alpha-fetoprotein (AFP)-producing hepatoblastoma in a 14-month-old girl are reported. Ten of 36 tumor cells were chromosomally abnormal, yielding the composite karyotype 47,XX,2q +, t(3;5)(p25;q31),dup(4)(q12q26), +20. Three of the chromosomal abnormalities (2q+, break at 4q12, +20) have been reported previously and might be of pathogenetic significance.     

Distribution of T-lymphocyte subpopulation in blood and spleen of normal cattle and cattle with enzootic bovine leukosis

Immunocytochemical and flow cytometry techniques were used to examine T-lymphocyte subpopulations in peripheral blood lymphocytes (PBLs) and spleen from cases of enzootic bovine leukosis (EBL) in adult cattle, and from normal cattle (adult and young), with a panel of monoclonal antibodies against bovine leucocyte differentiation molecules. Both in PBLs and spleen, the percentages of T-lymphocyte subpopulations (CD3+, CD4+, CD8+, and WC1 + gamma delta T lymphocytes) of EBL-affected and normal adult cattle were significantly lower than those of normal young cattle. The percentages of these T-lymphocyte subpopulations in the PBLs of adult cattle with EBL were lower than those of normal adult cattle, but the converse was true in the spleen. It is suggested that tumour immunity occurred in the spleen. Histological examination revealed no follicular hyperplasia in the spleen, and the proliferation of neoplastic cells began in the red pulp. It is concluded that the spleen is not the organ initially responsible for the transformation of EBL lymphoma and that neoplastic cells migrating from peripheral blood are metastatic.     

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.     

Chromosome aberrations in 35 primary ovarian carcinomas.

Cytogenetic analysis was performed on short-term cultures of primary ovarian carcinomas from 62 patients. Cytogenetic analysis was successful in 59 cases. Clonal chromosome aberrations were detected in 35 tumors. Only numerical changes or a single structural change were found in five carcinomas: trisomy 12 was the sole anomaly in two tumors, one tumor had the karyotype 50,XX, + 5, + 7, + 12, + 14, a fourth tumor had a balanced t(1;5), and the fifth tumor had an unbalanced t(8;15). The fact that four of these five carcinomas were well differentiated suggests that simple karyotypic changes are generally characteristic of these less aggressive ovarian tumors. The majority of the cytogenetically abnormal tumors (n = 30) had complex karyotypes, with both numerical and structural aberrations and often hypodiploid or near-triploid stemlines. The numerical imbalances (comparison with the nearest euploid number) were mostly losses, in order of decreasing frequency -17, -22, -13, -8, -X, and -14. The structural aberrations were mostly deletions and unbalanced translocations. Recurrent loss of genetic material affected chromosome arms 1p, 3p, 6q, and 11p. The breakpoints of the clonal structural abnormalities clustered to several chromosome bands and segments: 19p13, 11p13-15, 1q21-23, 1p36, 19q13, 3p12-13, and 6q21-23. The most consistent change (16 tumors) was a 19p + marker, and in 12 of the tumors the 19p + markers looked alike.