Techniques in Animal Cytogenetics (Principles and Practice)
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However, array-based techniques only identify imbalances resulting in deletions or duplications, so balanced rearrangements such as translocation, inversions, or rings may not be identified, and require a standard chromosome analysis.
Cytogenetic abnormalities are present in essentially all cases of plasma cell myeloma, and a subset of abnormalities has been associated with prognostic differences Table Classical cytogenetic studies demonstrate an abnormal karyotype in only one third of cases because of the low proliferative rate of plasma cells in vitro.
The demonstration of any karyotypic abnormality by metaphase cytogenetics is a surrogate marker of a relatively high proliferative rate and is associated with an adverse prognosis, irrespective of the specific abnormalities present. In recent years, FISH studies have emerged as the favored technique for cytogenetic analysis of plasma cell myeloma.
Indeed, the t 4;14 p16;q32 , which is associated with a poor prognosis, is cryptic by standard cytogenetics and is detected only by molecular diagnostic techniques. With some abnormalities, the prognostic impact varies with the detection technique employed. For example, chromosome 13 or 13q abnormalities have a greater prognostic significance when detected by metaphase cytogenetics than when detected by FISH. Therefore, FISH analysis is best interpreted with metaphase cytogenetics , and FISH panels, although valuable in the prognostic assessment of myeloma, are not an adequate substitute for classical cytogenetic analysis.
Grody, in Atlas of Hematopathology , Cytogenetics analyses can provide valuable and extremely relevant information to establish the presence of a malignant clone, determine the cell lineages in the disease process or clarify and confirm a diagnosis, provide prognostic predictive features, and monitor response to treatment and classification of neoplasms.
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The significance and usefulness of these will be discussed in the following chapters. A close relationship between the pathologist and the cytogeneticist is essential if maximum useful information is to be produced from the cytogenetics studies of hematological disorders.
Cytogenetics refers to both the study of the structure of chromosome material and the study of diseases caused by structural and numerical abnormalities of chromosomes. It includes the routine analysis of G-banded chromosomes, other cytogenetic banding techniques, and molecular cytogenetics such as fluorescent in situ hybridization FISH and comparative genomic hybridization CGH.
In human medicine, cytogenetics has played an important role in the diagnosis and clinical management of lymphomas, leukemias, and various solid tumors Campbell, ; Gebhart, ; Mundle and Sokolova, Both numerical and structural chromosome abnormalities have been identified in canine soft tissue sarcomas. Although many of the cytogenetic studies of animal tumors have been small case series, the advent and development of microarrays for canine CGH analysis will facilitate future studies.
In a small study of canine leukemia, cytogenetic abnormalities did not correlate with survival Grindem and Buoen, Similarly, a small study of feline leukemia found no relationship between cytogenetic abnormalities and prognosis Grindem and Buoen, In a study of the cytogenetics of canine lymphoma, dogs with tumors having trisomy 13 had a significantly longer median survival time compared to dogs with tumors having other chromosomal aberrations.
Further studies are needed to determine the prognostic importance of chromosomal aberrations in veterinary oncology. Anne Jouvet, in Practical Surgical Neuropathology , Cytogenetic studies on PCs are exceedingly rare and have not shown any clinically relevant nonrandomly distributed abnormalities. PCs present both numerical and structural aberrations in conventional cytogenetic studies, but the results have not been highly consistent.
In three conventional cytogenetic studies, the karyotypes of PCs were mostly pseudodiploid, and the common abnormalities included losses of chromosomes 4, 5, 14, and In two patients, loss of all or part of chromosomes 22, 11, and 1 was observed. Of note, in a recent comparative genomic hybridization CGH study, no gains or losses in PCs were found.
Techniques in Animal Cytogenetics : Bernard Dutrillaux :
Chromosome analysis will not identify all genetic anomalies. In addition, chromosomal analysis can be challenging in cancer cells, where morphologic preservation may be less than in somatic cells. Accuracy of chromosomal analysis is also heavily dependent on the skill of the technologist.
Lastly, mosaicism may be an important factor, as low level mosaicism may be missed. The number of cells routinely analyzed is dependent on the tissue type, whether the specimen has been cultured, and the purpose of the analysis.
Due to hypocellular bone marrow, it frequently occurs that there are insufficient metaphases for an adequate analysis. In such cases, FISH targeting specific anomalies should be considered. Most frequent anomalies in AA include trisomy 8, uniparental disomy of the 6p 6pUPD  , 5q-, anomalies of chromosome 7 and Abnormal cytogenetic clones often are small at diagnosis, and may arise during the course of the disease or may be transient and disappear after immunosuppression IST [13,14]. While abnormalities of chromosome 7 and 5 even in absence of dysplasia turn the diagnosis more likely in MDS, other cytogenetic findings are less categorical.
Del 13q has been reported in patients with MDS and other hematologic malignancies cases  as well as in patients with bone marrow failure syndrome. Patients presented with del 13q were reported mainly associated with good response to IST [16,17].
Hélène HAYES, Senior Research Scientist
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