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Simple Summary: Ki-67, a protein found in actively growing and dividing cancer cells, serves as an important tumor biomarker in breast cancer. A higher Ki-67 level indicates faster cell multiplication. In this study, pathologists evaluated Ki-67 using two methods: a visual whole-slide assessment and a quantitative tumor margin analysis. The study was performed in early-stage breast cancer patients. The findings revealed that Ki-67 is a reliable method for guiding clinical decision-making. Notably, a cut-off value of 20% appeared most appropriate for prognosis estimation and prediction of therapy benefit in our cohort of patients. Proliferation determined by Ki-67 immunohistochemistry has been proposed as a useful prognostic and predictive marker in breast cancer. However, the clinical validity of Ki-67 is questionable. In this study, Ki-67 was retrospectively evaluated by three pathologists using two methods: a visual assessment of the entire slide and a quantitative assessment of the tumour margin in 411 early-stage breast cancer patients with a median follow-up of 26.8 years. We found excellent agreement between the three pathologists for both methods. The risk of recurrence for Ki-67 was time-dependent, as the high proliferation group (Ki-67 ≥ 30%) had a higher risk of recurrence initially, but after 4.5 years the risk was higher in the low proliferation group. In estrogen receptor (ER)-positive patients, the intermediate Ki-67 group initially followed the high Ki-67 group, but eventually followed the low Ki-67 group. ER-positive pN0-1 patients with intermediate Ki-67 treated with endocrine therapy alone had a similar outcome to patients treated with chemotherapy. A cut-off value of 20% appeared to be most appropriate for distinguishing between the high and low Ki-67 groups. To summarize, a simple visual whole slide Ki-67 assessment turned out to be a reliable method for clinical decision-making in early breast cancer patients. We confirmed Ki-67 as an important prognostic and predictive biomarker. [ABSTRACT FROM AUTHOR]

Identification of underlying cytogenetic (CG) aberrancies plays a significant role in risk stratification of hematological malignancies. These abnormalities can be due to aberrancies that affect the number or structure of chromosomes. Numerical chromosomal abnormalities are called aneuploidies, which result from either gain or loss of whole chromosomes. Ploidy assessment by CG is a laborious and less sensitive technique. With the aid of fluorescent nucleic acid binding dyes, the total DNA content and different phases of the cell cycle specific to any population of interest can be deciphered and analyzed by flow cytometry (FCM). DNA index (DI), a parameter derived by FCM DNA analysis, is equivalent to conventional CG-based ploidy assessment. In this study, the technical aspects and implications of FCM DNA assessment among patients diagnosed with acute lymphoblastic leukemia and plasma cell myeloma are discussed. [ABSTRACT FROM AUTHOR]

Abstract Background About 7 million people die from various types of cancer every year representing nearly 12.5% of deaths worldwide. This fact raises the demand to develop new, effective anticancer, onco-suppressive, and chemoprotective agents for the future fighting of cancers. Genistein exhibits pleiotropic functions in cancer, metabolism, and inflammation. It functions as an antineoplastic agent through its effect on the cell cycle, apoptotic processes, angiogenesis, invasion, and metastasis. Aim of the study The current study aimed to study the genistein onco-suppressive effects during 7,12-dimethylbenz[a]anthracene (DMBA)-induced oral carcinogenesis in hamsters’ buccal pouch utilizing flow cytometry analysis (FMA), as a fast-diagnosing tool, in addition to the histopathology. Material and methods The buccal mucosa of adult male Syrian hamsters was painted with paraffin oil only (group 1), DMBA mixed in mineral oil (group 2), or orally administrated genistein along with painting DMBA (group 2B). The buccal mucosa was utilized for flow cytometric analysis and histopathological examination. Results Grossly, DMBA-induced carcinogenesis started at the 9th week. Progressive signs appeared in the following weeks reaching to large ulcerative oral masses and exophytic nodules at the 21st week. Histologically, invasive well-differentiated oral squamous cell carcinoma (OSCC) appeared in the underlying tissues from the 12th week, showing malignant criteria. Genistein had delayed clinicopathological change, which started 6 weeks later, than the DMBA-painted hamsters, as mild epithelial dysplastic changes. This became moderate during the last 6 weeks, without dysplastic changes. Flow cytometry revealed that DMBA led to considerable variation in DNA proliferation activity, aneuploid DNA pattern, in 47.22% of hamsters and significantly raised the S-phase fragment (SPF) values, which drastically reduced after genistein treatment. Conclusion Taken together, genistein could be employed as an onco-suppressive agent for carcinogenesis. Moreover, FMA could be used as an aiding fast tool for diagnosis of cancer.

To evaluate S-phase fraction as a predictor of invasiveness and cancer-specific survival in upper tract urothelial carcinoma (UTUC). One hundred and fifteen patients having undergone radical nephroureterectomy were analysed with histology in radical nephroureterectomy specimens as reference test and S-phase fraction as index test. Ploidy and S-phase were determined using flow cytometry. Differences in S-phase fraction were calculated between stages, grades (WHO 1999 and 2004 classifications), ploidy and patients that died of UTUC and those who did not. Five- and 10-year-cancer-specific survivals were calculated. Areas under the ROC curve (AUCs) of S-phase fraction in relation to tumour stage and to death from UTUC were measured. Multiple Cox regression was performed. Independent prognostic markers of death from UTUC were S-phase fraction and stage. Correlation between S-phase fraction and risk of dying from UTUC was strong, with a 17% greater risk of death from UTUC with every 1% increase in S-phase fraction, hazard ratio = 1.17, 95% CI = 1.10–1.25, p < 0.001, Spearman's rho ρ = 0.65. AUCs for S-phase fraction as predictors of stage and death from UTUC were 0.8 (95% CI = 0.705–0.894) and 0.77 (95% CI = 0.67–0.87), respectively. Cancer-specific survival was statistically significantly different between stages, ploidy and WHO 1999 grades, but not between WHO 2004 grades. This was also reflected in S-phase fraction, which differed in LG-G1 compared with LG-G2 and in HG-G2 compared with HG-G3. S-phase fraction was a good test for predicting both invasiveness and cancer-specific survival. Using both WHO 1999 and 2004 classifications, rather than one system alone, had a higher predictive value of cancer-specific survival. [ABSTRACT FROM AUTHOR]

Introduction: A rare entity of a B-cell malignancy with precursor B-cell phenotype and concomitant translocation t(8;14) or variant MYC translocation exists. These cases show clinical, pathological and molecular overlap between precursor B-lymphoblastic leukaemia or lymphoma and Burkitt leukaemia or lymphoma (BLL). Case presentation: We report a case from February 2019 at the Charlotte Maxeke Johannesburg Academic Hospital, South Africa, of a 9-month-old infant with a predominantly extracranial soft tissue mass showing extradural extension. There was no involvement of the peripheral blood or bone marrow. Fine needle aspiration and Tru-Cut biopsy of the soft tissue scalp mass showed the tumour to be of precursor B-cell phenotype. Contrastingly, an immunophenotypic assessment revealed a high S-phase fraction and raised concern for BLL. This prompted testing for the translocation t(8;14) by fluorescence in-situ hybridisation analysis, which confirmed this aberration. Management and outcome: Based on the published experience of other centres, the patient was initiated on a BLL protocol. Despite an excellent clinical response, the patient succumbed to neutropenic sepsis six months after diagnosis. Conclusion: Leukaemia or lymphoma with translocation t(8;14) or variant MYC translocation and precursor B-cell phenotype is a rare entity with a varied clinical presentation. This poses a challenge for diagnosis and classification and a clinical dilemma for the choice of treatment.

Background: FxCycleTM Violet (FCV) based flow cytometric (FCM) DNA ploidy analysis is a rapid and simple tool that can substantiate in characterizing the biological behaviour across the spectrum of haematological malignancies and correlates with cytogenetic studies. Materials and Methods: In this prospective study, we performed simultaneous immunophenotyping with FCV based on ploidy analysis in n=132 consecutive new samples, comprising n=110 samples of haemato-lymphoid neoplasms, including acute leukemias (n=67, 60.9%), CML with myeloid blast crisis (n=1, 0.9%), MDS with excess blasts (n=2, 1.8%), mature B cell/ T cell neoplasms (n=37, 33.7%), multiple myeloma (n=3, 2.7%) along with n=22 normal samples. The FCM DNA data was compared with corresponding conventional karyotyping results, wherever available. Results: In FCM ploidy analysis (n=110), the overall DNA index (DI) ranged from 0.81 to 2.17 and S-Phase fraction (SPF) from 0.1-31.6%. Diploidy was seen in n = 90 (81.8%), low-hyperdiploidy in n = 10 (9.1%), high-hyperdiploidy in n = 7 (6.4%) with one case each (0.9% each) having near-tetraploidy, high-hypodiploidy and low-hypodiploidy. The DI of all viable cell populations in normal samples ranged from 0.96-1.05. Conventional karyotyping was performed in n=76/110 cases (70%) with n= 11/76 (15%) culture failures. The modal chromosome number ranged from 45 to 63. A concordance of 95.4% (n=62/65) was noted with corresponding FCM DI. Conclusion: FCV-based ploidy is a sensitive technique that provides complementary information and ascertains a strong correlation with conventional cytogenetics across all haemato-lymphoid neoplasms. It can detect aneuploidy in all B-ALL and myeloma cases, even in hemodiluted samples with cytogenetic culture failure; supplement the diagnoses of erythroleukemia, and provide a useful screen for a higher grade lymph node disease in lymphoma cases with SPF > 3%.

Background: FxCycleTM Violet (FCV) based flow cytometric (FCM) DNA ploidy analysis is a rapid and simple tool that can substantiate in characterizing the biological behaviour across the spectrum of haematological malignancies and correlates with cytogenetic studies. Materials and Methods: In this prospective study, we performed simultaneous immunophenotyping with FCV based on ploidy analysis in n=132 consecutive new samples, comprising n=110 samples of haemato-lymphoid neoplasms, including acute leukemias (n=67, 60.9%), CML with myeloid blast crisis (n=1, 0.9%), MDS with excess blasts (n=2, 1.8%), mature B cell/ T cell neoplasms (n=37, 33.7%), multiple myeloma (n=3, 2.7%) along with n=22 normal samples. The FCM DNA data was compared with corresponding conventional karyotyping results, wherever available. Results: In FCM ploidy analysis (n=110), the overall DNA index (DI) ranged from 0.81 to 2.17 and S-Phase fraction (SPF) from 0.1-31.6%. Diploidy was seen in n = 90 (81.8%), low-hyperdiploidy in n = 10 (9.1%), highhyperdiploidy in n = 7 (6.4%) with one case each (0.9% each) having near-tetraploidy, high-hypodiploidy and low-hypodiploidy. The DI of all viable cell populations in normal samples ranged from 0.96-1.05. Conventional karyotyping was performed in n=76/110 cases (70%) with n= 11/76 (15%) culture failures. The modal chromosome number ranged from 45 to 63. A concordance of 95.4% (n=62/65) was noted with corresponding FCM DI. Conclusion: FCV-based ploidy is a sensitive technique that provides complementary information and ascertains a strong correlation with conventional cytogenetics across all haemato-lymphoid neoplasms. It can detect aneuploidy in all B-ALL and myeloma cases, even in hemodiluted samples with cytogenetic culture failure; supplement the diagnoses of erythroleukemia, and provide a useful screen for a higher grade lymph node disease in lymphoma cases with SPF > 3%. [ABSTRACT FROM AUTHOR]

Introduction: A rare entity of a B-cell malignancy with precursor B-cell phenotype and concomitant translocation t(8;14) or variant MYC translocation exists. These cases show clinical, pathological and molecular overlap between precursor B-lymphoblastic leukaemia or lymphoma and Burkitt leukaemia or lymphoma (BLL). Case presentation: We report a case from February 2019 at the Charlotte Maxeke Johannesburg Academic Hospital, South Africa, of a 9-month-old infant with a predominantly extracranial soft tissue mass showing extradural extension. There was no involvement of the peripheral blood or bone marrow. Fine needle aspiration and Tru-Cut biopsy of the soft tissue scalp mass showed the tumour to be of precursor B-cell phenotype. Contrastingly, an immunophenotypic assessment revealed a high S-phase fraction and raised concern for BLL. This prompted testing for the translocation t(8;14) by fluorescence in-situ hybridisation analysis, which confirmed this aberration. Management and outcome: Based on the published experience of other centres, the patient was initiated on a BLL protocol. Despite an excellent clinical response, the patient succumbed to neutropenic sepsis six months after diagnosis. Conclusion: Leukaemia or lymphoma with translocation t(8;14) or variant MYC translocation and precursor B-cell phenotype is a rare entity with a varied clinical presentation. This poses a challenge for diagnosis and classification and a clinical dilemma for the choice of treatment. [ABSTRACT FROM AUTHOR]

Endometrioid endometrial carcinoma is a cancer type with generally excellent prognosis when diagnosed at an early stage, but there is a subset of patients with relapsing disease in spite of early diagnosis and surgical treatment. There is a need to find prognostic markers to identify these patients with increased risk of relapse. Depth of myometrial invasion, histological grade, and presence of lymphovascular invasion are known risk factors. DNA content (ploidy) and proliferation measured as S-phase fraction (SPF) have been discussed as prognostic markers but need additional evaluation. We evaluated relapse-free survival (RFS) with respect to ploidy and SPF, which was analyzed by flow cytometry on fresh tumor tissue, in a cohort of 1001 women treated for stage I endometrioid endometrial carcinoma in northern Sweden during the period of 1993–2010, with a median follow up time of 12.0 years. Data were obtained from historical records. In simple analysis, both aneuploidy and high SPF were associated to increased risk of relapse with hazard ratios (HR) 2.37 (95% CI 1.52–3.70) and 1.94 (95% CI 1.24–3.02), respectively. Our data also confirmed stage, tumor grade, and ploidy as independent prognostic markers in an age adjusted cox regression multivariable analysis but we did not find SPF to contribute to prognosis. However, the combination of aneuploidy and high SPF identified a group of patients with increased risk of relapse, HR 2.02 (95% CI 1.19–3.44). In this study, which is the largest study of ploidy and SPF in stage I endometrioid endometrial carcinoma using fresh frozen tissue, aneuploidy was shown to be an independent prognostic marker. Furthermore, the combination of aneuploidy and high SPF could be used to identify patients with increased risk of relapse. [ABSTRACT FROM AUTHOR]

HER2 gene, located in chromosome 17q12–q21, encodes a transmembrane protein of 185 kDa with tyrosine kinase activity and an important role in tumorigenesis. The presence of the Ile655Val polymorphism identified in transmembrane domain of the HER2 protein was observed in tumors with different locations. Previous studies have shown that changing the existing isoleucine (Ile) to valine (Val) at codon 655 of the HER2 gene causes an intensification of dimerization, autophosphorylation of the HER2 receptor, as well as tyrosine kinase activity, which influences cell growth and tumor proliferation. The aim of this study was to assess the HER2 codon 655 polymorphism related to tumor cells proliferation in urinary bladder cancer. Tumors obtained from patients with bladder cancer were analyzed by PCR-RFLP for the presence of polymorphism at codon 655 of the HER2 gene. The expression of HER2 and pHER2 proteins was determined immunohistochemically. The DNA content, from the tumor cells, was analyzed by flow cytometry for cell cycle phases and DNA ploidy. HER2 heterozygous genotype was dominant in carcinomas with co-expression of HER2 and pHER2. The tumors HER2 and pHER2 positive from Ile/Val genotype group had a higher level of S-phase fraction than those from Ile/Ile genotype group. The S-phase fraction was also elevated in heterozygous than homozygous tumor in patients with lymph node metastases. The HER2 heterozygous cases were aneuploid predominantly. The presence of Ile/Val genotype in bladder carcinoma seems to be associated with a high proliferation. These results may suggest that HER2 codon 655 polymorphism can be used in the evaluation of disease progression in bladder cancer. [ABSTRACT FROM AUTHOR]

The potential prognostic significance of DNA flow cytometric measurements (DNA ploidy and S-phase fraction) in breast cancer remains in dispute. Inconclusive data, primarily due to the lack of consistent standardization and quality control programs, have limited its translation into clinical practice. The aim of the present review, based on the 25-year experience of the Portuguese Institute of Oncology of Lisbon, is to assess the clinical relevance and application of DNA flow cytometry for the prognosis of breast cancer. Overall, data from Portuguese Institute of Oncology of Lisbon indicate that DNA flow cytometry provides significant prognostic information that is biologically relevant and clinically useful for the management of patients with breast cancer. Furthermore, this data has demonstrated the independent value of DNA aneuploidy as a prognostic indicator of poor clinical outcome in various subgroups of patients with early or locally advanced breast cancer at short- and long-term follow-up. Notably, aneuploidy identifies subsets of patients with grade (G)1 or G2 tumours who exhibit a poor clinical outcome. These patients may benefit from adjuvant chemotherapy, particularly those with luminal A and luminal B/human epidermal growth factor-2-negative endocrine-responsive breast cancer. In conclusion, data from Portuguese Institute of Oncology of Lisbon reinforces the clinical importance and utility of DNA flow cytometric analysis, particularly DNA ploidy, in the prognostic assessment and therapeutic planning for patients with breast cancer. [ABSTRACT FROM AUTHOR]

Introduction: In early-stage endometrial carcinoma, there is controversy regarding the prognostic value of the flow cytometric variables DNA ploidy (diploid vs. aneuploid) and S-phase fraction. In Sweden, the former is included in national guidelines despite poor scientific support and the latter is not used clinically. This study investigates the prognostic properties of these variables, together with classical histopathological variables, in multivariate analysis in a stringently stratified material.Material and Methods: Consecutive, population-based patient material restricted to International Federation of Gynecology and Obstetrics (FIGO) 2009 stage I endometrioid endometrial carcinoma (n = 1140) was retrospectively collected from routinely reported data from medical records. Data on age, FIGO stage, degree of differentiation, S-phase fraction, DNA ploidy status, and adjuvant treatment were included in the study. Cumulative incidence curves with other causes of death as a competing risk were used for univariable analysis for the primary endpoint endometrial cancer death. Cox proportional hazards regression analysis was used for multivariate modeling of all endpoints, and for univariable analysis for the secondary endpoints overall survival and time to progression.Results: An S-phase fraction value of >5.5% was associated with worse outcome (for endometrial cancer death: hazard ratio 2.25; 95% CI 1.38-3.67; p = 0.001, adjusted) and DNA ploidy status was not, for all endpoints tested.Conclusions: In FIGO stage I endometrioid endometrial carcinoma, DNA ploidy status had no prognostic value, whereas the S-phase fraction may be used to identify those with a higher risk of adverse clinical outcome. [ABSTRACT FROM AUTHOR]

Background: Histopathological grading of oral dysplastic lesions is the method of choice for evaluating malignant and potentially malignant disorders. Owing to inter- and intra-observer variability, determination of the DNA ploidy status of lesions may serve as an adjunct in the prediction of malignant transformation. Aim: To correlate histopathological grading and ploidy status in potentially malignant and malignant disorders of the oral mucosa. Settings and Design: A pilot study was done with 30 patients (10 patients with oral potentially malignant disorders predominantly leukoplakia, 10 patients with oral malignant lesions and 10 patients with normal mucosa). Materials and Methods: Incisional biopsy was done after isolating the biopsy site with 1% Toluidine blue staining. Two sections of the tissue were removed and sent for histopathological and Flow-cytometric analysis respectively. Histopathological diagnosis was obtained and compared with Flow-cytometric results which were graded as diploid and aneuploid. Further, the S - phase fraction, DNA index were also calculated to evaluate the severity of malignant transformation or malignancy. Statistical Analysis: The results were analyzed using Pearson Chi-Square Test. Results: There exists a significant correlation between histopathology and ploidy status in both potentially malignant and malignant group. (P = 0.002). Conclusion: The data from this study has shown that DNA Ploidy analysis can be used as a valuable tool in assessing the carcinomatous progression of potentially malignant and malignant lesions. INSET: 75. [ABSTRACT FROM AUTHOR]

Background: Pathological detection of lymphovascular space invasion (LVSI) is a useful prognostic marker for patients with endometrial cancer. However, LVSI is criticized for its subjectivity and poor reproducibility. To improve the outcomes of patients with endometrial cancer, we evaluated objective parameters associated with LVSI to generate more accurate LVSI assessments and to identify patients with high-risk disease. Methods: We reviewed the medical records of 137 patients with endometrial cancer. Flow cytometry was used to determine DNA ploidy and S-phase fraction. Estrogen and progesterone receptor (ER and PR) levels and p53 and k-ras mutational status were tested. Results: LVSI was found in 36 patients (26.3%). Patients with LVSI had significantly decreased recurrence-free survival and overall survival compared to those without LVSI. Aneuploid tumors were significantly more frequent in LVSI-positive patients compared with LVSI-negative patients (odds ratio = 5.208, P < 0.001). With the exception of p53 mutational status, there was a statistically significant relationship between LVSI and other parameters tested. However, by multivariate analysis, DNA ploidy and S-phase fraction were significantly correlated with LVSI ( P = 0.034 and 0.001, respectively). Conclusion: Ploidy and S-phase fraction correlate with LVSI, which is a significant independent predictor of clinical outcome in patients with endometrial cancer. [ABSTRACT FROM AUTHOR]

Abstract: The aim of this study was to investigate the potential clinical utility of DNA flow cytometry biomarkers, ploidy, and S-phase fraction (SPF) in predicting overall survival in cervical cancer. This prospective study involved 159 patients with cervical carcinoma (median follow-up, 48 months). Pretreatment clinical staging was done according to the FIGO 2009 update classification. Biopsy tumor samples were used for flow cytometry analysis and histological examination. A prognostic study was performed using both Cox and Bayesian Weibull regression models. Eighty (50.3%) tumors presented DNA aneuploidy, mostly observed in adenosquamous (AS) cell carcinoma (8 of 9 cases) and adenocarcinoma (AC) (12 of 17 cases). The median SPF value (8.6%) was used for discriminating low vs. high tumor cell proliferation. High SPF significantly correlated with aneuploidy (p <0.001). All AS carcinomas had SPF>15%, while all ACs presented SPF<10% (p <0.001). Forty-three (27%) patients died of the disease during follow-up. Log-rank tests revealed significant differences between survival curves for older patients (≥44 years) (p =0.029), advanced clinical staging (p <0.001), and DNA diploidy in stage IIB of disease (p =0.039). Both regression analyses showed that advanced clinical staging and low SPF independently predict worse overall survival of patients. The results suggest that DNA flow cytometry parameters can provide additional predictive information in cervical cancer management. [Copyright &y& Elsevier]

Purpose: The quality of life in postmenopause is seriously affected by the symptoms related to vaginal atrophy. To evaluate in a 3-month, prospective, randomized, double blind, study whether vaginal suppositories containing genistein might improve genital symptoms, colposcopical and cytologic findings or modify DNA cytometric features in postmenopausal women affected by vaginal atrophy, in comparison with vaginal suppositories containing hyaluronic acid (HA). Methods: A total of 62 postmenopausal women were randomly assigned to receive intravaginally 97 μg of genistein (group A, n = 31) or 5 mg of HA (group B, n = 31) daily for 15 days continuously/month for 3 months. Vaginal and cervical smear, colposcopy, vaginal biopsy were performed before and at the end of the study. Maturation value (MV) was calculated. Flow cytometric analysis of DNA ploidy (DI) and S-phase fraction (SPF) were performed. Results: After 90 days of study, a significant improvement was obtained in genital symptoms, colposcopy scores and MV ( p < 0.001) in both groups; the improvement obtained by genistein was more effective especially regarding genital score ( p value between groups 0.001). No significant change was found in SPF value and DI. Conclusion: Both treatments improved genital symptoms, colposcopical features and MV, although genistein was more effective on genital score. Both treatments did not significantly influence flow cytometry parameters, although genistein showed slight decrease in DI, with a normalization of the aneuploid content present in some cases that could represent an additional application of intravaginal phytoestrogen therapy, providing an alternative therapy of vaginal atrophy in postmenopausal patients. The results of this investigation should be considered preliminary and need to be verified in larger, prospective studies. [ABSTRACT FROM AUTHOR]

Background: The risk of malignant conversion in oral leukoplakia is well documented. Histological findings are often unreliable and it is difficult to predict on the basis of clinical and histopathological changes which leukoplakic lesion will turn malignant. Methods: We used the technique of flow cytometry to evaluate the ploidy status, DNA index and S-phase fraction in leukoplakia, oral cancer and normal oral mucosal biopsies and compared it with histological findings. The study was carried out on 30 patients with oral cancer, 60 with leukoplakia and 30 with normal oral mucosal biopsies. Results: The aneuploidy rate in oral cancers was 64%, for leukoplakia 20%, while all normal mucosal biopsies were diploid. Aneuploid lesions also had a greater S-phase fraction (SPF). The DNA Index (DI) of aneuploid oral cancers was 1.72 and aneuploid leukoplakias was 1.24. Leukoplakia specimens which showed histological evidence of dysplasia had aneuploidy rate of 38%, DI of 1.19 and SPF of 6.2%. The corresponding values for leukoplakia specimens without dysplasia were 14%, 1.09 and 4.1%, respectively. Conclusion: The method of flow cytometry can be used to identify the subset of leukoplakia patients who are at a higher risk of malignant conversion. These patients could undergo more rigid surveillance or undergo excision biopsy of their lesions. [ABSTRACT FROM AUTHOR]

It is often difficult and challenging task to differentiate aplastic anemia (AA) from hypoplastic myelodysplastic syndrome (HMDS) among the patients with bone marrow aplasia. This is possibly because of the considerable clinical, cytological and histological similarities between these two disorders. As prognostic and therapeutic approach to AA and HMDS are different, it is imperative to differentiate them at the time of initial diagnosis. Various attempts have been made in the past to differentiate AA from HMDS. In the present study, we explored the value of certain new parameters i.e. S-phase fraction (SPF) and aneuploidy that could be used for this purpose. The study included 46 consecutive patients with aplastic anemia, 15 patients with HMDS along with 32 age and sex-matched control subjects. S-phase fraction and aneuploidy analysis was carried out by flow cytometry using Mod Fit-LT V3.0 software. The mean SPF value was significantly lower (p=0.02) in patients with AA and higher (p=0.01) in HMDS as compared to that of the control. Aneuploidy was present in 15.2% patients with AA and in 33.3% HMDS cases. During follow-up, 4 patients with AA developed MDS, out of these, three patients had aneuploidy as well as high SPF value at the time of diagnosis. Two patients with HMDS who had aneuploidy and high SPF, converted into AML. Eleven patients died during the study, in whom 8 had aneuploidy and high SPF value. We conclude that high SPF value and presence of aneuploidy favour the diagnosis of HMDS rather than AA. SPF and aneuploidy may be important parameters in patients with AA to predict their propensity to evolve into myelodysplastic syndrome and acute myeloid leukemia. SPF value may also be useful in the early diagnosis of HMDS before morphologically evidence of dysplasia is apparent. [ABSTRACT FROM AUTHOR]

Background: The prognostic significance of DNA ploidy and the S-phase fraction (SPF) have been extensively studied in breast cancer, but their clinical utility remains controversial. The type of tumour material can substantially influence flow cytometric DNA measurements. Material obtained by fine needle aspiration (FNA) biopsy is very suitable for flow cytometric DNA analysis because it contains a low proportion of non-tumour cells and less debris than tissue samples. Methods: The prognostic significance of DNA ploidy and SPF, determined on FNA samples, was analysed in 770 breast cancer patients, diagnosed between 1992 and 1997. DNA ploidy and SPF were determined at the time of diagnosis as part of the diagnostic work-up. The median follow-up was 90 months. Survival analysis included overall cancer specific survival (OS), disease free survival (DFS) and survival after recurrence (SAR). Other variables included in survival analyses were age, histological grade, histological type, lymph node status and tumour size. Disease free interval and the site of recurrence were also included in SAR analysis. Results: DNA ploidy and SPF correlated with tumour type, size, lymph node involvement and, especially, tumour grade. In a univariate analysis, both aneuploidy and high SPF were associated with shorter OS, DFS and SAR, but only SPF retained its independent prognostic significance in multivariate analyses. Independent prognostic variables for OS were node status, histological grade, SPF and tumour size. Node status, histological grade and SPF were independent predictors of DFS, while the site of recurrence, SPF, histological grade, disease free interval and age were independent predictors of SAR. Conclusions: DNA ploidy and SPF can be efficiently and routinely determined on FNA samples. High SPF is independently associated with a worse clinical outcome of patients with breast cancer. Although SPF and histological grade share prognostic information to some degree, SPF provides additional, less subjective prognostic information. The prognostic value of SPF determined on FNA samples could be even more relevant in neoadjuvant settings and for patients not amenable for surgical treatment, when histological grade cannot be assessed. [ABSTRACT FROM AUTHOR]

Histological assessment for prognostication of myxofibrosarcomas remains challenging. We previously reported independent prognostic value of Skp2, an oncoprotein promoting S-phase progression ( Clin Cancer Res 2006;12:487–98). We evaluated S-phase fraction (SPF) and ploidy of myxofibrosarcomas and the association between SPF and Skp2. Flow cytometric findings were analyzed for 75 cases and correlated with clinicopathological factors, Skp2 labeling index (LI), metastasis-free survival (MeFS), and overall survival (OS). Forty-seven and 28 cases were classified as diploid and nondiploid, respectively. High SPF (≥20%) was detected in 32 of 61 interpretable cases. Skp2 overexpression (LI ≥ 10%) was seen in 36 of 72 cases with scoring. Nondiploidy correlated with higher French Federation of Cancer Centers (FNCLCC) grades ( P = .006), remarkable necrosis ( P = .010), and Skp2 overexpression ( P = .018). Noticeably, SPF was significantly related to Skp2 LI ( P < .001, r = .458), FNCLCC grade, American Joint Committee on Cancer stage, and mitotic rate. Nondiploidy predicted shorter OS ( P = .0045) and MeFS ( P = .0489), whereas SPF ≥ 20% was only associated with inferior MeFS ( P = .0252). In multivariate analyses, nondiploidy independently correlated with both OS ( P = .020, RR = 3.337) and MeFS ( P = .013, RR = 5.780), together with Skp2 overexpression ( P = .014 for OS; P = .017 for MeFS) and disease-positive margins ( P = .004 for OS; P = .002 for MeFS). Skp2 promotes S-phase progression in myxofibrosarcomas. SPF provides no independent prognostic usefulness; it is probably overshadowed by Skp2. Nondiploidy adds another predictive value to Skp2 overexpression and disease-positive margins in prognostication. [ABSTRACT FROM AUTHOR]

Abstract: Objective: To investigate the correlation of DNA ploidy, S-phase fraction (SPF) and the expression of cyclin A to the clinical prognostic factors in patients with epithelial ovarian cancer. Study design: A prospective analysis was performed on 46 ovarian tumor patients. The DNA ploidy and SPF were determined by flow cytometry and the expression of cyclin A was measured by immunohistochemical staining. Results: Compared with benign and borderline tumors, the malignant tumors expressed higher levels of cyclin A (P =0.007), more aneuploid cells (P =0.018) and higher SPF (P =0.015). With regard to DNA ploidy and the clinical prognostic factors, aneuploid cells increased with tumor grade (P =0.011), the disease stage (P =0.009) and residual volume (P =0.001). When residual tumor was more than 2cm, the expression of cyclin A was increased (P =0.043). Three-year survival was low for patients with tumors expressing cyclin A in over 10% of cells (P =0.003). Conclusions: The data suggest that the assessment of the DNA ploidy, SPF and the expression of cyclin A may provide important information for predicting the prognosis of epithelial ovarian cancer. [Copyright &y& Elsevier]

B-Cell Non-Hodgkin's Lymphomas comprise a biological heterogeneous group of haematological neoplasms with variable clinical behaviour. For clinical and prognostic purposes their classification has been based on several important parameters such as DNA content measured by flow cytometry. The present work tries to analyse the results and the recommendations published in the "Consensus Review of the Clinical Utility of DNA Flow Cytometry in Neoplastic Hematopathology" (Maine, 1992), and the studies published thereafter. [ABSTRACT FROM AUTHOR]

Abstract: DNA ploidy studies on salivary gland tumours have shown that the proportion of aneuploid cases, although confined to the malignant entities, is considerably lower than for other solid malignancies. To analyse whether the S-phase fraction (SPF) may contribute to discrimination of diploid malignant from benign tumours, DNA flow cytometric data from 45 different malignant salivary gland tumours was compared with that of 121 pleomorphic adenomas. All benign tumours were diploid. Twelve malignant tumours contained aneuploid cell populations. The SPF values for diploid malignancies ranged between 0.9% and 11.0% (mean 3.9%), and between 0.5% and 7.9% (mean 2.7%) for pleomorphic adenomas. A 4% cut-off value gained statistical significance for discriminating diploid malignant tumours from pleomorphic adenomas (P <0.01). The sensitivity for SPF>4% was 46% and the positive predictive value was 40%. A sensitivity of 60% and a positive predictive value of 54% was achieved by combining aneuploidy and SPF>4%. These results show that DNA flow cytometry may contribute to diagnostic assessment in salivary gland tumours. [Copyright &y& Elsevier]

We previously developed an immunohistochemical method for estimating cell cycle state and phase in tissue samples, including biopsies that are too small for flow cytometry. We have used our technique to examine whether primary abnormalities of the cell cycle exist in laryngeal neoplasia. Antibodies against the markers of cell cycle entry, minichromosome maintenance protein-2 (Mcm-2) and Ki67, and putative markers of cell cycle phase, cyclin D1 (G1-phase), cyclin A (S-phase), cyclin B1 (G2-phase) and phosphohistone H3 (Mitosis) were applied to paraffin-embedded sections of normal larynx (n=8), laryngeal dysplasia (n=10) and laryngeal squamous cell carcinoma (n=10). Cells expressing each marker were determined as a percentage of total cells, termed the labelling index (LI), and as a percentage of Mcm-2-positive cells, termed the labelling fraction (LF). The frequency of coexpression of each putative phase marker was investigated by confocal microscopy. There was a correlation between Mcm-2 and Ki67 LIs (ρ=0.93) but Mcm-2 LIs were consistently higher. All cells expressing a phase marker coexpressed Mcm-2, whereas Ki67 was not expressed in a proportion of these cells. The putative phase markers showed little coexpression. Labelling index values increased on progression from normal larynx through laryngeal dysplasia to squamous cell carcinoma for Mcm-2 (P=0.001), Ki67 (P=0.0002), cyclin D1 (P=0.015), cyclin A (P=0.0001) and cyclin B1 (P=0.0004). There was no evidence of an increase in the LF for any phase marker. Immunohistochemistry can be used to estimate cell cycle state and phase in laryngeal biopsies. Our data argues against primary cell cycle phase abnormalities in laryngeal neoplasia.British Journal of Cancer (2006) 95, 314–321. doi:10.1038/sj.bjc.6603262 www.bjcancer.com Published online 11 July 2006 [ABSTRACT FROM AUTHOR]

Objective: The objective of our study is to evaluate prognostic significance of DNA quantification by flow cytometry in ovarian tumor.Methods: A prospective analysis was performed on 56 ovarian tumor patients treated in the Yonsei Medical Center from Feb. 2000 to Jan. 2003.Results: Regarding the association between tumor grade and the DNA quantitative analysis, as tumor grade increased, the quantity of aneuploid cells and S-phase fraction (SPF) increased. In addition, SPF was increased significantly in the advanced staged patients (P=0.04) and SPF was significantly increased in aneuploid tumors (P=0.03). The overall survival rate was poor for patients with aneuploid tumors and for patients with tumors showing over SPF (10%).Conclusion: Our data demonstrated that the prognosis was poor for patients with aneuploid cancers or increased SPF. Therefore, DNA quantification by flow cytometry may provide important information for predicting the prognosis of the disease. [ABSTRACT FROM AUTHOR]

DNA ploidy pattern and S-phase fraction (SPF) measured by flow cytometry and expression of the retinoblastoma gene product (pRB) estimated by immunohistochemistry were correlated with outcome in 114 patients who received a curative operation for primary non-small cell lung cancer (NSCLC). One hundred ten tumors yielded an adequate DNA histogram, and all tumors exhibited an assessable immunohistochemical stain. DNA diploidy was detected in 31 tumors and DNA aneuploidy in 79 tumors. The mean SPF was 14.1±6.4%. Eighty tumors were positively stained, and 34 tumors were negative for pRB. Multivariate analysis clarified that both TNM staging and DNA ploidy were prognostic factors after surgery. In 39 recurrent cases, the SPF value was inversely correlated with disease-free interval. With only supportive care after recurrence, high SPF tumors and pRB-negative tumors progressed rapidly, whereas active treatment yielded an equivalent effect on recurrent tumors regardless of the SPF or pRB expression. DNA ploidy is related to the risk of recurrence, while SPF is related to tumor growth rate and the impact of active treatment on recurrence. The utility of pRB expression was limited. The combination of DNA ploidy and SPF allows practical stratification of the biologic aggressiveness of NSCLC. [Copyright &y& Elsevier]

Purpose: To determine whether quantitative flow cytometric androgen receptor density expression (MFC ratio) in prostate cancer was associated with S-phase fraction.Methods: Flow cytometry was performed to determine DNA aneuploidy, S-phase fraction, percentage of androgen receptor (AR)-positive cells, and MFC ratio in prostate cancer patients.Results: MFC ratio showed distinct clustering. Eight patients had a low MFC ratio of 1.78-2.74, while 10 patients had high MFC ratios between 4.99 and 6.48. The S-phase fraction had average values of 11.05 vs 4.92 in tumors with high vs low MFC ratio (P<0.01).Conclusion: S-phase fraction was significantly higher in tumors with high AR density.Prostate Cancer and Prostatic Diseases (2003) 6, 294-300. doi:10.1038/sj.pcan.4500672 [ABSTRACT FROM AUTHOR]

We studied S-phase fraction (SPF) and aneuploidy in peripheral blood leucocytes of patients with chronic myeloid leukemia (CML) in chronic phase (CML-CP, n=41), accelerated phase (CML-AP, n=6), and control subjects (n=12) with an aim to find out their role in early detection of accelerated phase. The SPF and aneuploidy were studied through flow-cytometry using LT-Mod. Fit software. Mean SPF value in CML-AP (9.28±3.46%) and in CML-CP (4.76±2.30%) were significantly higher than in normal controls (0.28±0.21%), (P<0.005, P<0.001). CML-CP patients having higher SPF (>7%) converted to accelerated phase within 18 months of follow-up while those with lower SPF (<7%) did not. Aneuploidy was present in 34.14% of CML-CP and all patients of CML-AP whereas no control subjects showed aneuploidy. Among CML-CP patients having SPF >7%, 86% developed aneuploidy during follow-up as compared to 18.50% of CML-CP with less than 7% SPF. We conclude that peripheral blood SPF and aneuploidy could be important parameters for prediction of evolution to accelerated phase in CML patients. [Copyright &y& Elsevier]

Background: Female Taiwanese breast cancer patients are younger than their Western counterparts. This study examined the predictors of axillary lymph node metastases in Taiwanese women with T1 breast cancer. Methods: Data from 394 Taiwanese women with T1 invasive breast carcinoma were retrospectively reviewed. Results: The data contained 6 T1a, 51 T1b, and 337 T1c breast tumors. The patients’ ages ranged from 23 to 82 years (mean ± SD, 48.2 ± 11.4 years; median, 46.4 years). Axillary nodal metastases were present in 38.3% of the patients (16.7% in T1a, 35.3% in T1b, and 39.2% in T1c tumors). The patients with nodal metastases had significantly greater body weights and S-phase fractions than those without nodal metastases. Univariate analysis revealed that unfavorable pathology, lymphovascular invasion, S-phase fraction >7%, and nondiploid DNA ploidy were significantly associated with lymph node metastases. Lymphovascular invasion was the only significant variable as the independent predictor in the multiple logistic regression analysis. In the Cox proportional hazards regression analysis, axillary nodal status and lymphovascular invasion were significantly associated with survival. Conclusions: Taiwanese women with small breast cancer displayed a relatively higher incidence of axillary lymph node metastases than Western women. Axillary lymph node dissection or sentinel lymph node biopsy should be conducted on Taiwanese patients with small invasive breast carcinomas, particularly when risk factors exist. [ABSTRACT FROM AUTHOR]

Objective: To investigate the value of DNA ploidy and S-phase fraction (SPF) as factors that could predict response and outcome of invasive transitional cell carcinoma (TCC) of the bladder to neoadjuvant methotrexate, vinblastine, epirubicin and cisplatin (MVEC) chemotherapy. Materials and Methods: Twenty-four patients with localized invasive TCC of the bladder (stages T3 to T4a, NoMo) were treated with neoadjuvant MVEC chemotherapy. DNA flow cytometry was performed in paraffin-embedded tissue obtained by transurethral resection before chemotherapy. Radical cystectomy specimens were utilized for complete pathologic staging. The tumors were subdivided into high- and low-SPF tumors according to their SPF value. DNA ploidy status was evaluated into two groups (diploidy and nondiploidy). Results: DNA ploidy was not correlated to the response to chemotherapy (p = 0.67), and overall or disease-free survival (p = 0.27 and p = 0.69, respectively). Major response (pathologic complete response and partial response) was more often found in the high SPF group than among tumors with low SPF (p = 0.02). High SPF tumors were significantly associated with increased overall and disease-free survival (p = 0.0056 and p = 0.0059, respectively). Conclusion: A high SPF, but not DNA ploidy, may be helpful to identify patient likely to respond and survive longer following neoadjuvant MVEC chemotherapy in the treatment of invasive bladder cancer. [ABSTRACT FROM AUTHOR]

Objective: To investigate a possible relationship between DNA alterations in the "normal" residual mammary tissue of patients with breast cancer and survival. Design: Prospective study. Setting: University hospital, Spain. Subjects: 162 patients operated on for breast cancer between 1991 and 1995. Main outcome measures: Cytology, histopathology, optic karyometry, DNA ploidy, and S-phase fraction measured by flow cytometry in peritumoral and paratumoral tissue. Mortality, and univariate analysis. Results: DNA ploidy in peritumoral tissue was altered in 42 patients (26%), in 41 of whom the mean cytonuclear area was also altered. Of the 19 patients whose death within the study period was attributed to their cancer, 13 had peritumoral tissue in which both DNA ploidy and mean cytonuclear area were altered. On univariate analysis, there was significantly worse survival among the patients in whom both tumoral and peritumoral tissues were DNA aneuploid than among those in whom only tumoural tissue was DNA aneuploid (p < 0.0001). Conclusions: The presence of peritumoral or paratumoral tissue or both, with anomalous DNA content is associated with a reduced survival among women whose breast cancer has been treated by mastectomy. Additional studies including multivariate analysis are necessary to confirm our findings. [ABSTRACT FROM AUTHOR]

Treatment of nephroblastoma (Wilms' tumor) has presently achieved a 90% survival rate. Stage and grade are considered the most reliable prognostic parameters, but other biological factors are under study in order to improve patient stratification. Deoxyribonucleic acid (DNA) ploidy has been suggested to be useful in this setting. We retrospectively studied 79 patients with nephroblastoma (58 pretreated with chemotherapy and 21 not pretreated) by means of flow cytometry. DNA content and synthetic phase values were correlated with pathologic features and outcome. DNA modifications induced by chemotherapy were investigated. Sixty-nine tumors were diploid and 10 aneuploid. DNA content did not correlate with clinical course and was not modified by pretreatment. Aneuploid tumors were restricted to lower stages. Mean S-phase rate was lower and did not vary according to histology in pretreated tumors, while it was higher and increased with grade (p = 0.007) in previously untreated tumors. The fraction of cells in synthetic activity was related to outcome: Patients whose tumors displayed higher S-phase rates had a more favorable clinical course. Ploidy did not appear to be of prognostic significance. S-phase rate decreased after chemotherapy (p = 0.0002) and was related to survival. The worse outcome of pretreated patients might be attributed to a minor sensitivity to postoperative treatment: Preoperative chemotherapy would decrease the cell proliferation and might select resistant cellular clones of (possible) neoplastic residues. [ABSTRACT FROM AUTHOR]

To clarify the role of the interleukin (IL)-1/IL-1 receptor system in the progression of gastric carcinoma cells in patients with advanced gastric cancer, we measured the tissue concentrations of IL-1α, the expression of IL-1-receptor type I (IL-1RtI) on tumor cells, and the cell-growth activity through an analysis of DNA content. The concentrations of IL-1α were significantly higher in differentiated than in undifferentiated tumors ( P = 0.038). The expression of IL-1RtI was upregulated in the tumor cells associated with IFNγ, scirrhous type tumors, and T3 and T4 tumors. There was a clear linear correlation between the tissue concentrations of IL-1α and S-phase fractions in differentiated tumors ( r = 0.664, P = 0.003). Tumor cells with high IL-1α concentrations and low IL-1RtI expression had significantly greater S-phase fractions than those with low IL-1α concentrations, independent of IL-1RtI expression ( P = 0.024 in low IL-1RtI, P = 0.019 in high IL-1RtI). These findings indicate that IL-1α stimulates the growth of differentiated gastric carcinoma cells and that IL-1RtI expression is involved in tumor invasive activity. High S-phase levels were not necessarily associated with a high expression of IL-1RtI, which may be due to the downregulatory effects of high IL-1α concentrations. [ABSTRACT FROM AUTHOR]

Although several studies have examined brain tumor markers for prognostic value, few investigations have stratified analysis based on specific histologic grade. The objective of this study was to evaluate a single histologic grade of glioma, the grade IV glioma or glioblastoma (World Health Organization Classification), with a comprehensive panel of tumor markers in an attempt to identify those with prognostic significance. Tumor samples from a cohort of patients with glioblastoma multiforme ( n=32) were examined for tumor markers, DNA analysis, and clinical variables in an attempt to determine a ‘profile’ for this tumor. We used univariate and multivariate statistical analysis to determine the prognostic value of tumor cell ploidy, percent S-phase, DNA index, p53, and Ki-67 labeling index, as well as the variables of gender, race, age, location of tumor, history of chemotherapy, and primary versus recurrent tumor. Two additional tumor markers, multidrug resistance gene 1 and glutathione-S-transferase subtype pi, were included in the sample testing, but were not analyzed statistically. Univariate analysis indicated that increasing age had a strong association with decreased survival. Female gender, increasing Ki-67, no chemotherapy before sample collection, and primary glioblastoma showed some association with decreased survival in the univariate model. The univariate results indicated that race, side of tumor, ploidy, S-phase, DNA index, and p53 had no prognostic value. Multivariate modeling demonstrated that age, gender, and Ki-67 were the strongest factors associated with survival. The relevant literature is reviewed. [ABSTRACT FROM AUTHOR]

The role of circulating tumor markers in providing prognostic information has not been widely studied. In the current study, serum levels of the carbohydrate antigen 15-3 (CA 15-3) and carcinoembryonic antigen (CEA) were determined preoperatively in 364 breast cancer patients with no clinical signs of metastasis. The prognostic relevance of these markers for recurrence (175/364) and death of disease (104/175) was determined by Cox multivariate analysis, including the comparison with classical prognostic factors. High levels of both tumor markers were associated with aneuploid tumors with high S-phase fraction and high ornithine decarboxylase activity. CA 15-3 was highly associated with the number of positive lymph nodes and peritumoral lymphatic or blood vessel invasion. No significant associations were found between CEA or CA 15-3 levels and histologic grade, necrosis and steroid receptor status. In univariate analysis, preoperative values, using optimum cutoff values of CA 15-3 (40 U/ml) and CEA (6 ng/ml), were statistically significant for relapse-free survival and overall survival. In multivariate analysis, only node status, DNA ploidy and ornithine decarboxylase activity were independent predictors for relapse-free survival; the estrogen receptor status was a predictor of overall survival. In node-negative patients, ornithine decarboxylase activity was the only factor selected for relapse-free survival. In node-positive patients, the number of lymph nodes and DNA ploidy were the only variables selected for relapse-free survival or overall survival. Estrogen receptor and ornithine decarboxylase activity were excluded for relapse-free survival, but were significant prognostic factors for overall survival. Copyright © 2001 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Abstract: Proliferative activity has been proven to be of prognostic significance in breast carcinoma. This study was performed to compare the different proliferative fractions in the Egyptian population and to define the most suitable one for daily routine use in our surgical pathology laboratories. The proliferative activity of 63 invasive ductal carcinomas was evaluated by immunohistochemical staining of paraffin-embedded tissue sections with MIB-1 rabbit polyclonal antibody and the heat-induced epitope retrieval method, flow cytometric determination of the S-phase fraction (SPF) on frozen tissues, and estimation of the Scarff–Bloom–Richardson (SBR) grading and mitotic figure count (MFC) on hematoxylin and eosin-stained tissue sections. Fifty-two percent of invasive ductal carcinoma were aneuploid. The mean values of MIB-1 index, SPF, and MFC were 17.7 ± 12.3, 4.9 ± 3.8, and 5.2 ± 4.5, respectively, for diploid tumors; while for aneuploid tumors, they were 58.6 ± 31.9, 19.9 ± 12.2, and 23.1 ± 16.9, respectively. These values were significantly higher in aneuploid versus diploid tumors (p < 0.0001). A close correlation was found between MIB-1 index, SPF, MFC, and SBR grading (p < 0.0001). In conclusion, in surgical pathology laboratories that cannot afford the costs of flow cytometry and/or immunostaining, proper SBR grading and MFC can provide an estimation of the proliferation fraction similar to the flow cytometric SPF and MIB-1 immunostaining. [ABSTRACT FROM AUTHOR]

Background. Interleukin-1-alpha (IL-1α) produced by tumor cells stimulates the proliferation or the growth of several cancer cell lines. The aim of this study was to clarify the relationship between growth activity evaluated by DNA analysis and the concentration of tumor-derived IL-1α in gastric cancers in clinical cases. Methods. We measured the concentration of IL-1α in homogenized tumor samples obtained from 49 patients with gastric cancer, using an enzyme-linked immunosorbent assay, and we analyzed the cellular DNA content of paraffin-embedded tumor sections using flow cytometry. Results. Both the IL-1α concentration and the percentage of S-phase fraction were significantly correlated with liver metastasis, histologic type, pattern of tumor infiltration, quantity of stroma, and venous invasion. A good correlation was found between IL-1α concentration in tumors and the percentage of S-phase fraction ( R = 0.604, P < 0.0001). Conclusion. IL-1α may be related to the stimulation of cell growth of gastric cancer in clinical cases. [ABSTRACT FROM AUTHOR]

Flow cytometric DNA analysis with assessment of S-phase fraction and DNA ploidy was compared to Nottingham histologic grade. The study population consisted of 654 patients who presented between 1987 and 1996 with primary operable breast cancer and whose tumours had been analysed for S-phase fraction and DNA ploidy at the time of surgery. Grade, tumour size, node status, steroid receptor status, age, S-phase fraction and DNA ploidy were analysed univariately and multi-variately in a Cox proportional hazard analysis. In the univariate analyses all parameters were statistically significantly associated with breast cancer mortality during the follow-up period of 2–11 years. The most powerful predictor of death from breast cancer in the multiple regression analysis was grade. Patients with grade 1 tumours have excellent prognosis. We conclude that tumour grade is a strong prognostic indicator applicable to all breast cancer patients, regardless of size and nodal status, and advocate its general use. [ABSTRACT FROM AUTHOR]

From 1990 to 1996, 607 previously untreated, node-negative, invasive breast carcinomas were sampled by a pathologist for flow-cytometric DNA analysis. The aim of the present work was to study the correlations between flow cytometric results obtained thanks to the American Consensus (AC) guidelines of 1993 and the established clinico-pathological prognostic factors (T, grade, receptors), and despite a short global follow-up (mean of 4 years), to correlate flow cytometry with the outcome of the patients. In this study S-phase fraction (SPF) correlated strongly with tumor size, histological grade, lack of steroid receptors, histological type and was together with the mitotic activity a paramount prognostic factor even after multivariate analysis. This study compared also the technical criteria proposed by the AC with our own more stringent ones and concluded that the criteria of the AC are relevant and allow, thanks to the use of tertiles in the reporting of SPF values, a comparison of values obtained by different teams. Our review of the literature, focused on series using fresh material, enabled us to show that there is a rather wide agreement concerning the relationship between SPF and prognosis most often after multivariate analysis. This despite the lack of standardization in the design of the studies (implementation of the technical steps or reporting of results). When estimated from fresh or frozen material following AC’s guidelines, SPF along with mitotic activity should become a prognostic factor used in the daily practice by oncologists in the management of breast carcinomas. [ABSTRACT FROM AUTHOR]

A Swedish cooperative trial demonstrated that 5 years of adjuvant tamoxifen was more beneficial than 2 years of tamoxifen in the treatment of postmenopausal women with estrogen receptor (ER) positive, early stage, invasive breast cancer. The main aim of the present study was to investigate the importance of progesterone receptor (PgR) and ER concentration levels for patients participating in the trial and still distant recurrence free two years after the primary operation. Subgroup analyses revealed that only patients with ER positive and PgR positive breast cancer had improved distant recurrence free survival (DRFS) by prolonged tamoxifen therapy ( p=0.0016). Patients with ER negative and PgR negative as well as ER positive and PgR negative tumors showed no significant effect of prolonged tamoxifen ( p=0.53 and p=0.80, respectively). The percentage of ER negative and PgR positive breast cancers was too small (2.2%) for any meaningful subgroup analysis. There was a significant positive trend that the concentration level of PgR (high positive vs. low positive vs. negative) decreased the recurrence rate for those with prolonged therapy. No corresponding pattern was found for the ER content. S-phase fraction did not correlate to the recurrence rate of PgR positive breast cancers. Patients recurring during tamoxifen therapy had receptor negative tumors to a greater extent than those recurring after tamoxifen treatment. In conclusion, prolonged tamoxifen therapy for 5 years instead of 2 years was found to be beneficial for patients with ER positive and PgR positive breast cancer, whereas three extra years of tamoxifen had little or no effect for patients with ER positive but PgR negative tumors as well as for steroid receptor negative patients. [ABSTRACT FROM AUTHOR]

One hundred forty-three patients with gliomas of astrocytic origin (61 with glioblastomas multiforme (GM) and 82 with astrocytomas) received an intravenous infusion of bromodeoxyuridine (BUdR), 150-200 mg/m at the time of surgery, to label tumor cells undergoing DNA synthesis. BUdR-labeled cells were identified by the indirect immunoperoxidase method using anti-BUdR monoclonal antibodies. The percentage of BUdR-labeled cells, or BUdR labeling index (LI), was calculated by microscopic examination of selected viable areas of the tissue sections. The GMs had a median LI of 7.5%, and three quarters of these tumors had LIs greater than 5%. Highly anaplastic astrocytomas (HAAs) and moderately anaplastic astrocytomas (MoAAs) had median Lls of 2.3% and less than 1%, respectively. Among the HAAs, the Ll was 1% to 5% in 56% of tumors, greater than 5% in 26%, and less than 1% in 18%. More than 60% of MoAAs had LIs less than 1%, which agrees with the slow clinical progression of this type of tumor, and the remainder had LIs of 1.4% to 9.3%. These results show that histopathologically similar tumors may have different proliferative potentials. Measuring the proliferative potential of individual gliomas is therefore crucial for predicting the prognosis more accurately and for devising more tumor-specific treatment regimens for individual patients. [ABSTRACT FROM AUTHOR]

Cell proliferation of transitional cell bladder cancer (TCC) was determined by MIB-1 immunolabeling, volume-corrected mitotic index (M/V index) and S-phase fraction measurement in 207 patients with superficial (Ta-T1) bladder cancer. The results were compared to T category, WHO grade and DNA-ploidy. The MIB-1 score was related to T category ( P<0.001), WHO grade ( P<0.001), DNA ploidy ( P<0.0001), M/V index ( P<0.0001) and fraction of cells in S phase ( P<0.0001). The mean MIB-1 score was 6.37% for G1, 14.59% for G2 and 28.59% for G3 carcinomas ( P<0.001). The MIB-1 score for Ta tumors was 9.24% and for T1 tumors 25.34% ( P<0.001). The M/V index was 3.9 for G1, 11.5 for G2 and 25.9 for G3 tumors ( P<0.0001). The M/V index for Ta tumors was 6.4 and 25.3 for T1 tumors ( P<0.0001). WHO grade 1 tumors had 7.7%, grade 2 tumors 13.8% and grade 3 tumors 21.8% of cells in S phase ( P<0.001). Of grade 1 tumors, 97% were diploid and 3% aneuploid, and 78% of grade 2 tumors were diploid and 22% aneuploid. Of grade 3 tumors, 30% were diploid and 70% aneuploid ( P<0.001). Of Ta tumors, 92% were diploid and 8% aneuploid, respectively, whereas 40% of T1 tumors were diploid and 60% aneuploid ( P<0.0001). The results show that quantitative cell proliferation indices are associated with T category and WHO grade in superficial bladder cancer. The prognostic value of the S-phase fraction and mitotic index has been demonstrated in several previous analyses of prognostic factors while the value of MIB-1 score on bladder cancer prognosis remains to be established in further follow-up studies. [ABSTRACT FROM AUTHOR]

The bromodeoxyuridine (BrdU) labeling index was determined in 40 primary colorectal carcinomas by DNA flow cytometry using a BrdU-specific monoclonal antibody. The labeling index, or the fraction of cells in the S-phase of the cell cycle, ranged from 12% to 52%, with a mean of 28% (SEM, 2%). The labeling index in 19 patients was over 30%, which was termed a higher labeling index. There was no significant difference in the labeling index based on the clinical stage of the disease. During the 5-year follow-up after the apparently curative resection, 14 patients died of the disease, 1 died of an unrelated cause, 1 is alive with a recurrence of the disease, and 24 are alive without the disease. The higher labeling index was thus associated with a significantly poorer patient survival ( P=0.03 based on the generalized Wilcoxon test). The present study therefore disclosed that the S-phase fraction of tumor cells thus determined had prognostic significance in primary colorectal carcinoma. [ABSTRACT FROM AUTHOR]