Ploidy Pattern Research Articles

Histopathologic Diagnosis, Cell Cycle Parameters and Clinical Behavior of 90 Egyptian Brain Tumor Cases

This work aims at assessment of factors contributing to cell proliferation in relation to histopathologic diagnosis and clinical outcome of 90 brain tumour cases from Egypt. Cases were taken prospectively from the Neurosurgery Department of Mansoura University Hospitals, Egypt. Their median age was 46 years and their sex included 42 (46.7%) males and 48 (53.3%) females. Of these cases, 14 cases (15.6%) had an age <20 years. Brain biopsy samples were processed for histopathologic examination in addition to flow cytometeryic analysis of DNA ploidy pattern, apoptosis, p53 and Bcl2 expressions. Meningeal tumors were most frequent (37.8%) followed by astrocytic tumors (26.7%), sellar tumors (12.2%) while the neuroblastic tumors were detected in 10% of cases. Females were more affected by meningiomas and pituitary adenomas whereas males were more affected by astrocytic tumors. Older cases were affected mostly by meningeal and astrocytic tumors while the younger ones were more affected by neuroblastic tumors. Malignant tumors showed significant increased levels of mutant p53 expression, S phase of both diploid and aneuoploid cells than benign ones (P<0.05). On follow up, most of the cases affected with meningeal tumors had become symptom free while recurrence and death were mostly observed in astrocytic tumors. Significant increased expression of mutant p53 was also observed among recurrent cases (p<0.05) than cases that become free of symptoms. These results shows that cell cycle markers in addition to histopathology can help in predicting prognosis of brain tumours with a potential impact on management plan.

A study of DNA ploidy pattern, proliferation index and PCNA in duodenal carcinoma

12 cases of duodenal carcinoma were studied for nuclear DNA ploidy patterns, the proliferation index (PI), proliferating cell nuclear antigen (PCNA) positive score (1 = 0-25%, 2 = 26-50%, 3 = 51-75%, 4 = 76-100%) and PCNA positive rate. DNA aneuploidy was observed in 9 cases (75%) and PCNA staining was positive in 11 cases (91.6%). DNA ploidy patterns, PI, PCNA positive scores and positive rates were not related to each other. No relationship DNA ploidy patterns for PCNA positive scores and PCNA positive rates could be found. The relationship between PI and PCNA positive score was found not to be significant (P less than 0.10). PI was revealed to correlate significantly (P less than 0.05) to PCNA positive rate.

Low-grade gastric adenomas/dysplasias: Phenotypic expression, DNA ploidy pattern, and LOH at microsatellites linked to the APC gene

The adenoma–adenocarcinoma sequence in the colon is generally accepted. Regarding the stomach, however, opinions about tumorigenesis are different. We analyzed the phenotypes, DNA ploidy patterns, and microsatellite regions linked to adenomatous polyposis coli (APC) gene on chromosome 5q of low-grade gastric adenomas/dysplasias, Vienna Category 3, to investigate whether these lesions have the potential to become adenocarcinomas. Phenotypes were determined by immunohistochemical staining with monoclonal antibodies MUC2, MUC5AC, MUC6, CD10, and Cdx2. DNA ploidy patterns were determined by static cytofluorometry. Microsatellite analyses were performed using Genescan on genetic analyzer ABI PRISM 310. None of the 15 cases showed coexisting high-grade adenomas/dysplasias or adenocarcinoma. Eighty percent of the cases had the complete intestinal phenotype, and the remainder showed slight MUC6 positivity for the intestinal phenotype. All cases were positive for Cdx2 and showed diploid DNA. In addition, 46.7% of the cases exhibited loss of heterozygosity (LOH) of chromosome 5q. Except for one case, 5q-LOH was detected at a single locus in cases with the complete intestinal phenotype. According to previous studies, most gastric adenocarcinomas have the gastric phenotype and no 5q-LOH. These results suggest that low-grade gastric adenomas/dysplasias, Vienna category 3, seldom progress to gastric adenocarcinomas of the differentiated type.

Nuclear vs. plastid data: complex Pleistocene history of a circumpolar key species

To fully understand the contemporary genetic structure of plants, both nuclear and plastid markers are needed. Three chloroplast DNA (cpDNA) lineages, which probably diverged before the major Pleistocene glaciations, have been identified in the circumpolar/circumboreal Vaccinium uliginosum. Here we investigate its nuclear DNA variation using nuclear ribosomal internal transcribed spacer (ITS) sequences, DNA ploidy level measurements and amplified fragment length polymorphisms (AFLPs). We also extend the cpDNA dataset. Two ITS lineages, corresponding to diploids and tetraploids, respectively, were identified. However, both main sequence types apparently occurred in most individual plants but showed ploidy-biased homogenization and possibly reflect paralogy predating the origin of V. uliginosum. The ploidy levels were largely consistent with the cpDNA lineages, suggesting that the initial cpDNA divergence followed early polyploidizations. Five main AFLP groups were identified, consistent with recent glacial refugia in Beringia, western Siberia, the southern European mountains and areas south/east of the Scandinavian and Laurentide ice sheets. Except from the southern European mountains, there has been extensive expansion from all refugia, resulting in several contact zones. Surprisingly, the presumably older ploidy and cpDNA patterns were partly inconsistent with the main AFLP groups and more consistent with AFLP subgroups. A likely major driver causing the inconsistencies is recent nuclear gene flow via unreduced pollen from diploids to tetraploids. This may prevent cytoplasmic introgression and result in overlayed patterns formed by processes dominating at different time scales. The data also suggest more recent polyploidizations, as well as several chloroplast capture events, further complicating this scenario. This study highlights the importance of combining different marker systems to unravel intraspecific histories.

A model describing cell polyploidization in tissues of growing fruit as related to cessation of cell proliferation

Endoreduplication is a phenomenon, widespread among plants, which consists of an incomplete cell cycle without mitosis and leads to the increase of the nuclear DNA content. In this work, a model was developed describing cell proliferation and DNA endoreduplication over the whole fruit development, from the pre-anthesis period until maturation. In each mitotic cycle of duration tau, the proportion of cells proceeding through division depends on a constant parameter rho and on the progressive decline of the proliferating capacity . The non-dividing cells may either stop the reduplication fully, or switch to repeated syntheses of DNA without cell division, resulting in cell endoreduplication. A single constant parameter sigma describes the proportion of cells that moves from one to the next class of DNA content after each lapse of time tauE, considered to be the minimum time required for an endocycle. The model calculates the total number of cells and their distribution among eight classes of ploidy level. The dynamic patterns of cell proliferation and ploidy were compared with those obtained experimentally on two contrasting tomato genotypes. The approach developed in this model should allow the future integration of new knowledge concerning the genetic and environmental control of the switch from complete to incomplete cell cycle.

Prognostic cytogenetic markers in childhood acute lymphoblastic leukemia: Cases from Mansoura Egypt

The objective of the work was to evaluate children with acute lymphoblastic leukemia (ALL) showing resistance to immediate induction chemotherapy in relation to conventional and advanced cytogenetic analysis. The study was conducted on 63 ALL children (40 males and 23 females) with age range 4.5 months–16 years (mean = 7.76 years). They included 37 cases who attained a true remission and 26 complicated by failure of remission, early relapse or death. They were subjected to history, clinical examination and investigations including CBC, BM examination, karyotyping, FISH for translocations and flowcytometry for immunophenotyping and minimal residual disease diagnosis.Cases aged <5 years; male sex with organomegaly had better remission although statistically insignificant. Initially low HB <8 gm/dl, high WBCs and platelet counts >50.000/mm3 also showed better but non-significant remission rates. Most of our cases were L2 with better remission compared to other immunophenotypes. About 40 informative karyotypes were subdivided into 15 hypodiploid, 10 pseudodiploid, 8 normal diploid and 7 hyperdiploid cases; the best remission rates were noticed among the most frequent ploidy patterns. Chromosomes 9, 11 and 22 were the most frequently involved by structural aberrations followed by chromosomes 5, 12 and 17. Resistance was noted with aberrations not encountered among remission group; deletions involving chromosomes 2p, 3q, 10p and 12q; translocations involving chromosome 5; trisomies of chromosomes 16 and 21; monosomies of 5 and X and inversions of 5 and 11. Our conclusions were that cytogenetic and molecular characterizations of childhood ALL could add prognostic criteria for proper therapy allocation.

Prognostic cytogenetic markers in childhood acute lymphoblastic leukemia

To evaluate children with acute lymphoblastic leukemia (ALL) showing resistance to immediate induction chemotherapy in relation to conventional and advanced cytogenetic analysis. This work was conducted on 63 ALL children (40 males and 23 females) with age range 4.5 months-16 years (mean = 7.76 years). They included 37 cases attained true remission and 26 complicated by failure of remission, early relapse or death. They were subjected to history, clinical examination and investigations including CBC, BM examination, karyotyping, FISH for translocations and flowcytometry for immunophenotyping and minimal residual disease diagnosis. Cases aged 50.000/mm3 also showed better but non-significant remission rates. Most of the present cases were L2 with better remission compared to other immunophenotypes. Forty informative karyotypes were subdivided into 15 hypodiploid, 10 pseudodiploid, 8 normal diploid and 7 hyperdiploid cases; the best remission rates were noticed among the most frequent ploidy patterns. Chromosomes 9, 11 and 22 were the most frequently involved by structural aberrations followed by chromosomes 5, 12 and 17. Resistance was noted with aberrations not encountered among remission group; deletions involving chromosomes 2p, 3q, 10p and 12q; translocations involving chromosome 5; trisomies of chromosomes 16 and 21; monosomies of 5 and X and inversions of 5 and 11. Some cytogenetic and molecular characterizations of childhood ALL could add prognostic criteria for proper therapy allocation.

Cáncer renal incidental en pacientes menores de 40 años: hallazgos clínicos e histopatológicos

Objectiveto evaluate the clinical and pathological renal cancer (CR) characteristics in our series of tumours, analyzing its impact in the group of age less than 40 years Material and methodsWe studied 294 patients with CR. The pathologic characteristics were analyzed and DNA ploidy pattern of the surgical pieces were done in 252 patients. The patients were divided in two groups based on age, greater or less to 40 years, well then clinical and pathologic characteristics were compared between ResultsOf all patients, 26 of 294 patients (8,94%) were included in the young age group (less to 40 years). We did not found differences between both groups comparing stage, tumoral volume, treatment realized or DNA ploidy pattern, but in nuclear grade with more aggressive tumours in young people (p=0,0018), without differences in recurrence-free survival or actuarial disease specific survival rate ConclusionsThe findings in our study indicate that the natural history and outcome of the RC is similar in both older and younger patients. Therefore, in our opinion, the management of CR in young people should be established with independence of the age

Prognostic value of flow cytometry in surgically treated primary gastric lymphoma

To investigate whether flow cytometry could help to define the optimal therapeutic strategy of primary gastric lymphomas. Retrospective study of 46 patients having primary gastric lymphoma--according to Dawson criteria--in Ann Arbor stage IE and IIE, who were surgically treated. From selected paraffin-embedded tissue blocks of the tumor, DNA content was studied by flow cytometry (FC). Other pathological tumor features were analysed by hematoxiline-eosine and Giemsa stains as well as immunohistochemical study; any possible influence on postoperative survival was investigated through statistical analysis. The DNA ploidy pattern was diploid in 40 cases (87%) and aneuploid (hyperdiploid) in 6 (13%). Postoperative survival probability (PSP) was 62.7% at 5 years. Statistical analysis showed significant prognostic value for Ann Arbor classification--with higher PSP for stage IE (p = 0.009)--and FC parameters: diploid tumors had higher PSP than aneuploid tumors. Also tumors having S-phase (p = 0.044) or G2-M phase values (p = 0.023) under the respective mean values had higher PSP. No influence on PSP was found for wall invasion, Helicobacter pylori infection, Isaacson's histologic type or resection margin involvement. No significant relationship was appreciated between Isaacson's histologic type and DNA ploidy patterns. FC could be useful in assessing gastric lymphoma prognosis.

ANEUPLOIDY, A SOURCE OF GENETIC DIVERSITY FOR FRUIT SPECIES

Aneuploidy is reported among several plant species. Issued from open-pollinated polyploid plants and from targeted breeding, most genotypes can be saved after embryo culture that prevents plant failure due to abortion and growth abnormalities respectively. Aneuploid apple material revealed genetically stable through in vitro axillary branching and crossable after ex vitro grafting. However, certain genetic variability occurred after adventitious budding, spontaneously inducing polyploidisation and cytochimerism. Some cytochimerical clones probably sectorial chimeras, reversed to genetically uniform aneuploids after subcultures by axillary branching. Multiapexing and adventitious budding applied to these chimeras led to the regeneration of new genotypes characterized by stable and uniform ploidy or aneuploidy and mixoploidy as well. Different ploidy patterns of regenerants resulted from the hormonal balance of the shoot induction medium. The combination of the aneuploidy with the techniques of in vitro regeneration led to create new genotypes with a limited genetic variability. These are currently used for genetic analysis and the production of seedless fruit. It appears obvious that with the use of specific molecular markers, this way of genetic improvement could support the progression of innovative breeding programs.

Prognostic cytogenetic markers in childhood acute lymphoblastic leukemia: Cases from Mansoura, Egypt

Objective: To evaluate children with acute lymphoblastic leukemia (ALL) showing resistance to immediate induction chemotherapy in relation to conventional and advanced cytogenetic analysis.Subjects and methods: This work was conducted on 63 ALL children (40 males and 23 females) with age range 4.5 months–16 years (mean = 7.76 years). They included 37 cases who attained true remission and 26 complicated by failure of remission, early relapse or death. They were subjected to history, clinical examination and investigations including CBC, BM examination, karyotyping, FISH for translocations and flow cytometry for immunophenotyping and minimal residual disease diagnosis.Results: Cases aged <5 years; male sex with organomegaly had better remission although statistically insignificant. Initially low Hb < 8 gm/dl, high WBCs and platelet counts > 50,000/mm3 also showed better but non-significant remission rates. Most of our cases were L2 with better remission compared to other immunophenotypes. Forty informative karyotypes were subdivided into 15 hypodiploid, 10 pseudodiploid, 8 normal diploid and 7 hyperdiploid cases; the best remission rates were noticed among the most frequent ploidy patterns. Chromosomes 9, 11 and 22 were the most frequently involved by structural aberrations followed by chromosomes 5, 12 and 17. Resistance was noted with aberrations not encountered among remission group; deletions involving chromosomes 2p, 3q, 10p and 12q; translocations involving chromosome 5; trisomies of chromosomes 16 and 21; monosomies of 5 and X and inversions of 5 and 11.Conclusions: Cytogenetic and molecular characterizations of childhood ALL may add prognostic criteria for optimal therapy allocation.

Randomised study of adjuvant chemotherapy for completely resected p-stage I–IIIA non-small cell lung cancer

We evaluated the therapeutic usefulness of adjuvant chemotherapy in patients with completely resected non-small cell lung cancer (NSCLC). We also examined the relation between DNA ploidy pattern and the response to chemotherapy. A total of 267 patients with NSCLC (pathologically documented stage I, II, or IIIA) underwent complete resection, and DNA ploidy pattern was analysed. Patients with stage I disease (n=172) were randomly assigned to receive surgery alone (group A) or surgery followed by adjuvant chemotherapy (UFT (oral anti-cancer drug, a combination of Uracil and Tegaful) 400 mg day−1 for 1 year after surgery; group B). Stage II or IIIA disease patients (n=95) were randomly assigned to surgery alone (group C) or surgery followed by chemotherapy (two 28-day courses of cisplatin 80 mg m−2 on day 1 plus vindesine 3 mg m−2 on days 1 and 8, followed by UFT 400 mg day−1 for at least 1 year; group D). Eight-year overall survival rate in patients with stage I disease was 74.2% (95% confidence interval (CI): 64.4–84.0%) in group B and 57.6% (95% CI: 46.4–68.8%) in group A (P=0.045 by log-rank test). In patients with stage II and IIIA disease, no difference was found between groups C and D. Analysis according to DNA ploidy pattern revealed no difference between the groups. Postoperative chemotherapy with UFT was suggested to be useful in patients with completely resected stage I NSCLC. No difference was seen in relation to DNA pattern in any treatment group.

Correlation of protein expression, Gleason score and DNA ploidy in prostate cancer

The prognosis of prostate cancer correlates with tumor differentiation. Gleason score and DNA ploidy are two prognostic factors that correlate with prognosis. We analyzed differences in protein expression in prostate cancer of high and low aggressiveness according to these measures. From 35 prostatectomy specimens, 29 cancer samples and 10 benign samples were harvested by scraping cells from cut surfaces. DNA ploidy was assessed by image cytometry. Protein preparations from cell suspensions were examined by 2-DE. Protein spots that differed quantitatively between sample groups were identified by MS fingerprinting of tryptic fragments and MS/MS sequence analysis. We found 39 protein spots with expression levels that were raised or lowered in correlation with Gleason score and/or DNA ploidy pattern (31 overexpressed in high-malignant cancer, 8 underexpressed). Of these, 30 were identified by MS. Among overexpressed proteins were heat-shock, structural and membrane proteins and enzymes involved in gene silencing, protein synthesis/degradation, mitochondrial protein import (metaxin 2), detoxification (GST-pi) and energy metabolism. Stroma-associated proteins were generally underexpressed. The protein expression of prostate cancer correlates with tumor differentiation. Potential prognostic markers may be found among proteins that are differentially expressed and the clinical value of these should be validated.

Ovarian carcinomas with neuroendocrine differentiation: Review of five cases referring to immunohistochemical characterization

To review five ovarian carcinomas with varying degrees of neuroendocrine differentiation (ND) using an immunohistochemical study focused on the relationship with morphological features. ND was immunohistochemically analyzed using 21 antibodies by an indirect immunoperoxidase method, and ploidy pattern was analyzed using paraffin sections. The tumors were divided according to tumor cell size into 'small-sized' for case 1, 'intermediate-sized' for cases 2 and 3, and 'large-sized' for cases 4 and 5. Expressions of neuroendocrine markers and argyrophil reaction tended to be strengthened as tumor cell size increased. Cases 1, 2 and 3 showed diploid pattern and cases 4 and 5 showed aneuploid pattern. ND of ovarian carcinomas is closely related to morphological features represented by the cell size. Therefore, ovarian carcinomas with ND should be defined because the disease entity is not successfully integrated irrespective of the highly malignant potential.

Relationship between DNA Ploidy and Proliferative Cell Nuclear Antigen Index in Canine Hemangiopericytoma

The mitotic index is reported to be correlated with recurrence, mean patient survival, and metastasis of canine hemangiopericytoma (CHP). However, to the authors' knowledge, studies investigating the parameters that can predict recurrence or metastasis of CHP with low mitotic index have not been done. To evaluate growth kinetics of CHP with low mitotic index, a retrospective analysis of the proliferative activity by antiproliferative cell nuclear antigen monoclonal antibody and DNA contents by flow cytometry (FCM) was performed with 21 formalin-fixed and paraffin-embedded CHP samples. Of the 21 tumors evaluated by FCM, 6 (26.6%) were aneuploid tumors, and 15 (71.4%) were diploid tumors. There was significant correlation between the PCNA index and ploidy pattern. The diploid group had 39.1 +/- 9.2 PCNA index, whereas the aneuploid group's proliferative cell nuclear antigen (PCNA) index was 63.1 +/- 8.2. The diploid group had mean mitotic index value of 1.140 +/- 0.855, and the aneuploid group had a mean value of 1.067 +/- 0.767. From these results, the CHP samples with low mitotic index were classified into either the aneuploid group with higher PCNA index or the diploid group with lower PCNA index, suggesting that DNA ploidy and proliferative activity may give an indication about malignancy of CHPs with a low mitotic index.

Cáncer renal incidental en pacientes de edad geriátrica: hallazgos clínicos e histopatológicos

to evaluate the clinical and pathological characteristics of the CCR in our series of tumors, analyzing its impact in the group of age greater than 65 years. 300 patients with renal adenocarcinoma (CCR) were studied. In 252, ploidy pattern of DNA and pathologic characteristics of the surgical pieces were done. According to the criteria of the Spanish Society of Geriatrics, the patients were divided in two groups based on the age, greater and smaller of 65 years. The clinical and pathologic characteristics were compared between both groups. 103 of the 300 patients (33.3%) were included in the geriatric group. We did not found differences between both groups comparing stage, tumoral volume or treatment realized, but found differences in DNA ploidy pattern, recurrences and survey. The RCC in the elderly has a few clinical and pathological characteristics similar to the rest of patients in our series. Nevertheless differences exist in the average and actuarial survival, which is minor in the patient of more than 65 years, cause the percentage of detected aneuploidies and number of recurrences in not confined tumors in this group of age.

Correlation between expression of sialosyl-T antigen and survival in patients with gastric cancer.

Intestinal metaplasia of the gastric mucosa is an important lesion in the pathogenesis of gastric cancer. The expression of a mucin-associated antigen, sialosyl-T antigen, in the tissue of gastric cancer was investigated immunohistochemically to assess its biological role. Sialosyl-T antigen, detected by the monoclonal antibody TKH2, was present in 30 (23.6 per cent) of 127 lesions, and expressed sporadically in intestinal metaplasia. Patients negative for sialosyl-T had a significantly better 5-year survival than those who were positive (70.4 per cent versus 47.2 per cent). Grading by sialosyl-T antigen expression and the DNA ploidy pattern had a predictive value for prognosis in patients with gastric cancer. The biological role of sialosyl-T antigen is not clear but the mucin alteration reflects upon the biological behaviour of gastric cancer.

Prognostic value of DNA ploidy in primary gastric leiomyosarcoma.

The prognostic value of DNA ploidy was evaluated in 25 patients with primary gastric leiomyosarcoma, using paraffin-embedded archival specimens analysed by DNA flow cytometry. Ploidy patterns were diploid in ten tumours and non-diploid in 15. They did not correlate with clinicopathological features of the lesions such as tumour size and grade or mitotic index. A non-diploid pattern was associated with a significantly reduced patient survival rate (P < 0.05), as was higher tumour grade (P < 0.01). Neither tumour size nor mitotic index correlated with patient survival.

DNA ploidy pattern and tumour spread in gastric cancer.

The DNA ploidy pattern of gastric cancer was studied in 58 patients to investigate the heterogeneity between primary tumour and metastases. In both primary tumours and lymph node metastases, diploid patterns accounted for 33 per cent, whereas all liver metastases were aneuploid. The percentage of polyploid cells was higher in the liver metastases than in primary tumours and lymph node metastases. When the heterogeneity of DNA ploidy pattern between primary tumour and metastasis was evaluated, diploid tumours had a significantly lower rate of lymph node metastasis heterogeneity than aneuploid tumours. When the DNA ploidy pattern and survival were evaluated, the patients who had a diploid pattern in both primary tumour and metastasis had a significantly higher survival rate than the patients who had an aneuploid pattern in the primary tumour and metastasis (57 per cent versus 26 per cent at 5 years). These data suggest that cell heterogeneity is a common phenomenon in gastric cancer, and this may be important in the evolution of the disease. Furthermore, the role of the DNA ploidy pattern as a prognostic factor is emphasized.

Proliferation characteristics of human colorectal carcinomas measured in vivo

The cell proliferation kinetics of 100 human colonic and rectal adenocarcinomas have been studied in vivo by bromodeoxyuridine infusion and multiparameter flow cytometry. A total of 97 patients, three with synchronous tumours, consented to receive a single bolus dose of 250 mg between 2.4 and 16 h before curative or palliative surgery. By this method, the ploidy pattern, the total and aneuploid labelling indices (LI), the S phase duration (Ts) and the potential doubling time (Tpot) can be estimated. Of the tumours 48 were diploid and 52 were aneuploid. The mean and median total LI of 100 tumours were 9.0 per cent (range 0.7-22.2 per cent). The mean aneuploid LI was 12.1 per cent (median 12.0 per cent, range 2.0-25.5 per cent), and was significantly higher than the total LI (P = 0.01). The labelling index alone is not a sufficient indicator of proliferation, because the Ts also varies within and between tumours. The intertumour range of the Ts varied from 4.0 to 28.6 h. The mean was 14.1 h and the median was 13.1 h. The mean Tpot was 5.9 days (median 3.9 days) with a range of 1.7-21.4 days. No correlation was found between any kinetic parameters and the Dukes' classification or histological classification. The correlation between proliferation and prognosis will be established in due course.