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48 fine needle aspiration biopsy (FNAB) samples from 25 breast cancer cases, originally used for cytodiagnosis were subjected to DNA cytometry. There were air dried smears stained with the MGG method, and samples stained with HE or PAP stain after 50% ethanol fixation and cytocentrifugation. Different sampling strategies were applied. Four methods were tested: method 1: cell groups measured, method 2: all cells measured, method 3: free cells measured, and method 4: atypical free cells measured. Method 4 showed most often DNA aneuploid histogram patterns, sampling method 1 had the highest number of DNA diploid histogram patterns. Diagnostic approaches may benefit from a sampling method detecting the hiding aneuploid cell population. Grading of neoplasm could potentially benefit from other approaches.

Colorectal cancer is third leading cause of cancer mortality. About 60% of patients had already developed metastasis at the time of diagnosis. Vascular endothelial growth factor (VEGF) is crucial for the development of neovascularization and hence metastasis. This study aimed at investigating the relation between the expression of VEGF in biopsies from surgically dissected colon cancer and the survival of those patients. Biopsies were collected from 86 patients with advanced colon cancer and sections were stained by immunohistochemistry for VEGF. Patients received chemotherapy after the operation and were followed up for disease progression and survival. The clinical data were statistically analyzed with respect to the immunohistochemistry results. The survival of the patients was significantly longer in the patients for whom biopsies showed negative or weak expression of VEGF in comparison to those with moderate to high expression (p-value = 0.04). The expression of VEGF was more frequent in the patients who died as a consequence of the disease in comparison to the 10-year survivors. In conclusion, VEGF could be related to the survival of the patients with colorectal carcinoma and should be considered as a predictor of the prognosis.

Aim: To assess the predictive and prognostic value of neurofibromin (NF) expression in colorectal carcinoma (CRC)., Materials and Methods: The present series consists of archival samples from 191 patients with stage I, II, III, or IV CRC treated between 1981 and 1990 at the Turku University Hospital (Finland). Tumor biopsies as microarray blocks were analyzed for expression of NF by immunohistochemistry. Different grading systems were tested for NF expression., Results: A significant correlation between NF expression and tumor localization was found, with tumors arising in the colon showing intense NF expression more often than those arising in the rectum (p=0.014). Higher expression of NF was more common in tumors not responding to treatment (p=0.004). Tumors with multiple metastases showed higher expression of NF than those with single metastasis only (p=0.025). Furthermore, NF expression showed a borderline (p=0.068) correlation with gender; tumors of women showed higher NF expression that those of males. On the other hand, NF expression was not significantly associated with tumor recurrence, age, lymph node involvement, tumor grade and tumor stage or disease outcome., Conclusion: Positive NF expression in CRC is a sign of aggressive disease and poor outcome., (Copyright© 2016 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.)

Background: Despite compelling evidence from the genetic background and clinical studies indicating that cyclooxygenase-2 (COX2) up-regulation is a key step in carcinogenesis of colorectal carcinoma (CRC), controversy regarding its role as a prognostic factor exists. However, all evidence indicates that increased COX2 activity promotes progression of CRC. This study, aimed to evaluate the expression of COX2 in CRC, and correlate it with different patient clinicopathological data, emphasizing on the role of COX2 as a prognostic factor for CRC., Materials and Methods: In the present study, archival samples from 145 patients with stage I, II, III, or IV CRC treated during 1981-1990 at the Turku University Hospital (Finland) were used (as microarray blocks) to analyze COX2 expression by immunohistochemistry (IHC)., Results: Higher levels of COX2 expression were associated with higher TNM class (p<0.06), and higher Dukes' stage (p<0.045). In contrast, there was no significant correlation with age, gender, tumor grade or lymph node status. However, univariate survival analysis of metastases showed borderline association with COX2 expression in that patients with metastases with COX2-positive tumors were alive for shorter periods of time compared with patients whose tumors had no COX2 expression (p<0.023, log-rank)., Conclusion: COX-2 expression has shown a significant correlation with tumor stage and hence is assumed to be a prognostic factor in our cohort of colorectal cancer patients.

Background: We evaluated the relation of proliferative indices with clinicopathological features and prognosis in breast cancer (BC) of Libyan female patients. The data were compared with corresponding results in Finland and Nigeria., Patients and Methods: Histological samples of breast cancer from 130 patients were retrospectively studied. Mitotic activity index (MAI) and standardized mitotic index (SMI) were estimated., Results: There were statistically significant correlations between the proliferative indices and most clinicopathological features, with the strongest association observed for histological grade (P = 0.01 for SMI and P = 0.003 for MAI). The proliferative differences between Libyan, Nigerian, and Finnish population were prominent. The mean values of SMI and MAI in Libyan BC patients were 32.1 mitotic figures per square millimeter and 27.3 mitotic figures per 10 high-power fields, respectively. This is clearly lower than those in Nigeria but much higher than those in Finland. The differences between countries are seen in whole material and are also present in subgroups. The results indicated that mitotic activities can be reliable prognostic indicators in Libyan BCs, as they were among Finnish and Nigerian females. Univariate and multivariate analyses found at cut-offs of 19 and 44 mitosis/mm(2) of SMI were the most significant prognostic factors., Conclusions: Proliferative indices with careful estimation of the MAI and SMI could be applied as quantitative criteria for Libyan BC to separate the patients into good, moderate, and bad prognosis groups.

Aims: To study the diagnosis delay and its impact on stage of disease among women with breast cancer on Libya., Methods: 200 women, aged 22 to 75 years with breast cancer diagnosed during 2008-2009 were interviewed about the period from the first symptoms to the final histological diagnosis of breast cancer. This period (diagnosis time) was categorized into 3 periods: <3 months, 3-6 months, and >6 months. If diagnosis time was longer than 3 months, the diagnosis was considered delayed (diagnosis delay). Consultation time was the time taken to visit the general practitioner after the first symptoms. Retrospective preclinical and clinical data were collected on a form (questionnaire) during an interview with each patient and from medical records., Results: The median of diagnosis time was 7.5 months. Only 30.0% of patients were diagnosed within 3 months after symptoms. 14% of patients were diagnosed within 3-6 months and 56% within a period longer than 6 months. A number of factors predicted diagnosis delay: Symptoms were not considered serious in 27% of patients. Alternative therapy (therapy not associated with cancer) was applied in 13.0% of the patients. Fear and shame prevented the visit to the doctor in 10% and 4.5% of patients, respectively. Inappropriate reassurance that the lump was benign was an important reason for prolongation of the diagnosis time. Diagnosis delay was associated with initial breast symptom(s) that did not include a lump (p < 0.0001), with women who did not report monthly self examination (p < 0.0001), with old age (p = 0.004), with illiteracy (p = 0.009), with history of benign fibrocystic disease (p = 0.029) and with women who had used oral contraceptive pills longer than 5 years (p = 0.043). At the time of diagnosis, the clinical stage distribution was as follows: 9.0% stage I, 25.5% stage II, 54.0% stage III and 11.5% stage IV.Diagnosis delay was associated with bigger tumour size (p <0.0001), with positive lymph nodes (N2, N3; p < 0.0001), with high incidence of late clinical stages (p < 0.0001), and with metastatic disease (p < 0.0001)., Conclusions: Diagnosis delay is very serious problem in Libya. Diagnosis delay was associated with complex interactions between several factors and with advanced stages. There is a need for improving breast cancer awareness and training of general practitioners to reduce breast cancer mortality by promoting early detection. The treatment guidelines should pay more attention to the early phases of breast cancer. Especially, guidelines for good practices in managing detectable of tumors are necessary.

Background: The features of Libyan patients with breast cancer have not been fully investigated. The aim of this study was to evaluate the expression patterns of estrogen (ER) and progesterone receptor (PR), as well as nuclear morphometric features, in patients with breast cancer, and to correlate them with clinicopathological features and prognosis., Patients and Methods: Data for a total of 62 female Libyan patients with breast cancer, diagnosed between 2000 and 2006, were retrospectively studied. Their clinical and pathological data were collected and analysed. Immunohistochemical evaluation of ER and PR expression was also performed. Further more nuclear morphometry was carried out., Results: Of the 62 patients, disease in 10 was of the lobular type, 43 had invasive ductal and 9 had other carcinoma types; 47 out of 62 had lymph node involvement. Positive hormonal receptor expression was more common among those with lymph node-negative than lymph node-positive tumours. ER- and PR-positive patients appeared to have a better survival than ER- and PR-negative patients. The most significant difference, with respect to survival, was found between those bearing tumors with completely negative hormonal staining (J score 0) and those with positive staining (J score 1, 2 and 3). Larger nuclear size was associated with lymph node involvement and high-grade tumours (p<0.01 and p<0.0001, respectively), with shorter survival, larger tumour size and higher stage., Conclusion: The cut-off points for defining the groups with good or worse prognosis might be set, between score 0 and 1 (corresponding to 1% or fewer positive cells). Patients with ER- and PR-positive cancer had better overall survival than patients with hormonal receptor-negative cancer. In our hospital setting, ER and PR expressions and mean nuclear area (MNA) in breast carcinoma may be prognostically useful markers in guiding future treatment in prospective studies.

Background: We evaluated the relationship of the apoptotic activity index (AI) and the standardized mitotic-apoptotic ratio (SMI/AI) with clinicopathological features and prognosis in Libyan female breast cancer (BC) patients. We then compared our results with corresponding results in Finnish and Nigerian female BC patients., Methods: Histological samples of breast carcinoma from 130 patients were retrospectively studied: an estimation of the apoptotic activity per square millimeter (expressed as apoptotic activity index (AI)), and standardized mitotic-apoptotic ratio (SMI/AI) was made, and the results compared with the clinicopathological features and the patient's survival., Results: There was a statistically significant correlation between the AI and most of the clinicopathological features; the strongest association was observed for clinical stage lymph node (LN) status (P = 0.005). There were also correlations between AI and histological grade (P = 0.035), large tumor size (P = 0.011) and the clinical stage (P = 0.009). There were, however, prominent AI differences between Libyan, Nigerian and Finnish populations. The mean values of AI and SMI/AI in Libyan BC patients were 12.8 apoptotic figures per square millimeter and 2.8, respectively. The Libyan AI is slightly higher than in Nigeria, but much higher than in Finland. The differences between countries are seen throughout the samples as well as being present in certain subgroups. The survival analysis indicated that short survival time was associated with high apoptotic indices values and so can identify aggressive tumors and provide significant prognostic support. The cutoff (4 and 18 apoptosis/mm2) of AI might be applied as a quantitative criterion for Libyan BC to separate the patients into good, moderate and bad prognosis groups., Conclusions: The results indicated that the differences in AI among the three countries may be due to the known variation in the distribution of genetic markers in these populations. Improvement in health care and introduction of screening programs, however, could be very helpful in the Libyan population.

Background: We evaluated the association of the immunohistochemical Ki-67 expression, and S-phase fraction with clinicopathological variables and patient outcome., Patients and Methods: Histological samples from 100 primary Libyan breast carcinoma patients were retrospectively studied with monoclonal antibody to Ki-67. S-phase fraction was determined by DNA image cytometry., Results: The median Ki-67 percentage for all tumors was 27.5%, ranging from 1 to 80% and the median S-phase fraction (SPF) was 11%, ranging from 0 to 62 %. Tumors with high Ki-67 expression were found in 76% of patients and with high SPF values in 56%. Ki-67 expression was more frequent in tumors with high SPF than low SPF. High Ki-67 and high SPF were associated with advanced stages, poor differentiation of tumors, positive lymph nodes, and distant metastasis. The Ki-67 was associated with hormone receptor negative tumors. The SPF was higher in young patients (<50 years) than in older patients. In the overall population (median follow-up 49 months), patients with high Ki-67 and high SPF had shorter survival time and predicted recurrence than patients with low Ki-67 and low SPF. In a Cox multivariate analysis, high SPF (p= 0.007), hormonal status (p= 0.001) and clinical stage (p=0.005) were independent predictors of disease-specific survival. The Ki-67 (p=0.065) in borderline significance proved to be independent predictor of disease-free survival. The SPF showed more statistically significance with a high grade of malignancy and survival time than Ki-67., Conclusions: The SPF value is useful cell proliferation marker to assess tumor prognosis. These markers may reflect the aggressive behavior of Libyan breast cancer and predict of the recurrence. It is therefore important to take these markers into consideration to select a high risk subgroup of the patients for intensive treatment.

The present study evaluated the incidence of breast cancer in Libya and described the clinicopathological and demographic features. These features were then compared with corresponding data from patients from sub-Saharan Africa (Nigeria) and Europe (Finland). The study consisted of 234 patients with breast carcinoma, admitted to the African Oncology Institute in Sabratha, Libya, during the years 2002-2006. The pathological features were collected from pathology reports, patient histories from hospital files and the Sabratha Cancer Registry. The demographic differences between the Libyan, Nigerian and Finnish populations were prominent. The mean age of breast cancer patients in Libya was 46 years which was almost identical to that of Nigeria, but much lower than that of Finland. The Libyan breast cancer incidence was evaluated as 18.8 per 100,000 female individuals. This incidence was markedly higher in Finland, but was also high in Nigeria. Libyan and Nigerian breast cancer is predominantly of premenopausal type and exhibits unfavorable characteristics such as high histological grade and stage, large tumor size and frequent lymph node metastases. However, the histological types and histopathological risk features show similar importance regarding survival as European breast cancer cases. Survival in Libya ranks between the rates of survival in Nigeria (lowest) and Finland (highest). In conclusion, in Libya and other African countries, premenopausal breast cancer is more common than postmenopausal breast cancer. However, the opposite is true for Europe. Population differences may be involved, as suggested by the known variation, in the distribution of genetic markers in these populations. Different types of environmental impacts, however, cannot be excluded.

Background: Quantitative methods in combination with other objective prognostic criteria can improve the evaluation of a cancer patient's prognosis, and possibly predict response to therapy. One of the important prognostic and predictive markers is the mitotic count, which has proven valuable in many aspects. In this study, the prognostic value of the mitotic count was assessed in breast cancer (BC) patients in Saudi Arabia., Patients and Methods: The study comprised a series of 87 patients diagnosed and treated for breast cancer at the Departments of Surgery and Oncology, King Abdul-Aziz University Hospital, between 2000 and 2008. Mitotic counts were carried out using a standard laboratory microscope (objective, × 40; field diameter, 420 μm). The number of mitotic figures in 10 consecutive high-power fields (hpf) from the most cellular area of the sample gave the mitotic activity index (MAI, mitotic figures/10 hpf). The standardized mitotic index (SMI) recorded the mitotic count as the number of mitotic figures by area of the neoplastic tissue in the microscopic field, thus the number of mitoses in 10 consecutive fields was corrected for the volume fraction and field size (mitotic figures/mm²)., Results: The means of MAI and SMI of the tumors in the entire series of 87 patients were 15 mitotic figures/10 hpf (range 4-45) and 4 mitotic figures/mm² (range 1-9), respectively. The mitotic counts were higher in advanced stages than in early cancer (p < 0.04). The mitotic counts were significantly larger in patients with high-grade tumor (p < 0.004) and in cases with tumor metastasis (p < 0.004). The mitotic counts were also significantly larger in the recurrent cases than in non-recurrent ones (p < 0.02)., Conclusion: The quantitatively measurable mitotic counts of cancer cell nuclei are of significant prognostic value in invasive ductal carcinoma of the breast in Saudi Arabia and the mean cut-off values of MAI and SMI can be applied as objective (quantitative) criteria to distinguish breast cancer patients into groups with favorable and less favorable prognosis.

Background: The role of nuclear morphometry as a prognostic factor in breast cancer is well documented. The aim of this study was to evaluate this role in breast cancer in Saudi patients and to compare it with the experience in some African and European studies., Patients and Methods: Primary tumors from 135 patients were analyzed using an image overlay drawing system (Prodit Morphometry Program), for the following nuclear features: area, perimeter, diameter, and roundness., Results: The mean nuclear area (NA) was 93 microm(2) (range 45-168 microm(2)). The values of NA were higher in lymph node-positive patients than lymph node-negative patients and in advanced stages than early cancer. NA was significantly larger in patients with high grade tumor (p<0.0001) and in cases with tumor invasion (p<0.01). NA also was significantly larger in recurrent cases (103 microm(2)) than in non-recurrent ones (91 microm(2)). In univariate (Kaplan-Meier) analysis, NA was a significant predictor of disease-free survival (DFS) (log rank p<0.01), but not disease-specific survival (DSS). In multivariate (Cox) survival analysis, NA lost its significance as an independent predictor; response to treatment (p=0.0001) and tumor grade (p=0.030) being the only predictors of DFS. In a similar analysis for DSS, recurrence (p=0.040) and stage (p=0.003) were the only independent predictors., Conclusion: Nuclear morphometric profiles are helpful in identifying aggressive tumor phenotype (i.e. cases at risk for recurrence). The cut-off (93 mum(2)) of NA might be applied as quantitative criterion for Saudi female breast cancer to separate patients into good and poor prognosis groups. Mean NA of Saudi patients was markedly higher than the reported mean NA in the other studies and these differences might be due to technical variations or genetic bases.

Background: Defects of some DNA polymerases have shown cancer associations, but there are only limited data on DNA polymerase (Pol) epsilon., Materials and Methods: We examined 26 human brain neoplasm DNA samples and 8 control blood samples (from Poland) for possible mutations in the entire coding region of the 55 kDa small subunit of human DNA Pol epsilon gene using polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis, and sequence analysis of DNA., Results: One single base intronic transition in intron 14 was found. The AATT deletion previously found in some breast and colorectal tumors was not found in samples from brain neoplasms or controls, but it was found in 1/100 normal blood samples from South-West Finland., Conclusion: We found no evidence that potential mutations in the 55 kDa subunit of DNA Pol epsilon are a contributing factor in the development of the tested cases of human intracranial tumors.

Background: The sensitivity for identification of malignant cells in conventional fine-needle aspiration biopsy (FNAB) investigation is about 80%. This percentage is dependent on the number of examined cells, type of breast cancer, and experience of the examiner. The aim of our study was to estimate the supporting value of image DNA cytometry of FNAB of the breast, and do so by using different sampling methods., Materials and Methods: This retrospective study was based on 41 cases with an available histological diagnosis: 18 benign lesions and 23 malignant tumours were examined. The smears were submitted to image DNA analysis in a three-step protocol: (i) smears stained with HE method were destained and (ii) then restained with Feulgen staining for DNA and (iii) finally analysed using image cytometry., Results: All non-malignant cases had diploid histogram. However, a few of them had one or two cells of >5c category. Most histologically malignant cases were aneuploid. Only three invasive ductal carcinomas showed diploid histograms. All samples with aneuploid histograms were malignant., Conclusion: The results confirm earlier published data in the Finnish population and indicate that image DNA cytometric analysis of nuclear content is a useful marker for identification of malignant cells in FNAB, especially after free cell sampling. The method can be used to increase the cytological sensitivity and specificity in doubtful breast lesions.

Background: Defects of some DNA polymerases have shown associations with cancer, but data on DNA polymerase epsilon are limited. This study investigated mutations in the 55 kDa subunit gene of DNA polymerase epsilon in colorectal cancer., Materials and Methods: DNA from 16 human colorectal cancer and 9 control samples was studied with polymerase chain reaction-single-strand comformation polymorphism analysis and DNA sequencing., Results: DNA polymerase epsilon gene alterations were identified in 5 out of the 16 cases (31.2%). Two samples showed a T-C transition at exon 17 (potential tyrosine to histidine substitution), and an A-G transition at intron 7; one sample showed an A-G transition at intron 8. An AATT deletion was observed at intron 18 in 3 out of the 16 colon cancer cases (grades 2, 3, and 2, and Dukes' classes C, D, and C, respectively)., Conclusion: Because the AATT deletion has also been found in breast cancer, the region may be a mutation hot spot, possibly involved in the carcinogenetic path in advanced colorectal cancer.

Background: The cell cycle is promoted by activation of cyclin dependent kinases (Cdks), which are regulated positively by cyclins and negatively by Cdk inhibitors. Proliferation of carcinoma is associated with altered regulation of the cell cycle. Little is known on the combined alterations of cyclins A, B1, D1 and E in breast cancer in relation to the tumour grade and other prognostic factors., Findings: Immunohistochemical analysis of cyclins A, B1, D1 and E, estrogen receptor, progesterone receptor, Ki-67, Her-2/neu and CK5/6 was performed on 53 breast carcinomas. mRNA levels of the cyclins were analysed of 12 samples by RT-PCR. The expression of cyclins A, B1 and E correlated with each other, while cyclin D1 correlated with none of these cyclins. Cyclins A, B1 and E showed association with tumour grade, Her-2/neu and Ki-67. Cyclin E had a negative correlation with hormone receptors and a positive correlation with triple negative carcinomas. Cyclin D1 had a positive correlation with ER, PR and non-basal breast carcinomas., Conclusion: Cyclin A, B1 and E overexpression correlates to grade, Ki-67 and Her2/neu expression. Overexpression of cyclin D1 has a positive correlation with receptor status and non-basal carcinomas suggesting that cyclin D1 expression might be a marker of good prognosis. Combined analysis of cyclins indicates that cyclin A, B and E expression is similarly regulated, while other factors regulate cyclin D1 expression. The results suggest that the combined immunoreactivity of cyclins A, B1, D and E might be a useful prognostic factor in breast cancer.

Background: Approximately 30% of all colorectal cancer (CRC) patients are diagnosed with stage II disease. Adjuvant therapy is not widely recommended. However, it is well established that a subgroup of patients with stage II are at high risk for recurrence within their lifetime and should be considered for adjuvant chemotherapy. The present work was designed to assess the value of group IIA phospholipase A2 (PLA2) as a predictor of disease outcome in stage II CRC patients with long-term follow-up., Patients and Methods: The present study comprises a series of 116 patients who underwent bowel resection for stage II CRC during 1981-1990 at Turku University Hospital. Archival paraffin-embedded CRC tissue samples were used to prepare tissue microarray blocks for immunohistochemical staining with PLA2., Results: Fifty-five percent of all tumors were positive for PLA2. There was no significant correlation between PLA2 expression and age, sex, depth of invasion and lymph node status. In Kaplan-Meier survival analysis, there was a significant (P = 0.010) difference in disease-free survival (DFS) between patients with negative tumors (longer DFS) and those with positive tumors. The same was true with disease-specific survival (DSS), patients with PLA2-negative tumors living significantly longer (P = 0.025). In multivariate (Cox) survival analysis, however, PLA2 was not an independent predictor of DFS or DSS. In subgroup analysis, the right-sided tumors with negative PLA2 staining had remarkably better prognosis (P = 0.010) than PLA2-positive left-sided tumors., Conclusions: Quantification of PLA2 expression seems to provide valuable prognostic information in stage II CRC, particularly in selecting the patients at high risk for recurrent disease who might benefit from adjuvant therapy.

Background: The relation of nuclear morphometry measurements with clinicopathological features was evaluated along with prognosis in invasive female breast carcinoma in Libyan patients. Data was compared with corresponding results on Finnish, and Nigerian female breast cancer patients., Patients and Methods: Histological samples from 131 patients of breast carcinoma were retrospectively studied by computerized nuclear morphometry. In each case, 50 nuclei were measured and the mean nuclear morphometric features were calculated and compared with different clinicopathological features, and patient's survival., Results: There was statistically significant correlation between the mean nuclear area (MNA) and most clinicopathological features, with the strongest association observed for nuclear grade (p<0.0001). There was also correlation between nuclear area and tumor stage (p<0.04), tumor size (p<0.03) and lymph node (LN) status (p<0.001). A corresponding relationship was found between other size related features and clinical factors. The univariate analysis and survival analysis indicated that short survival time was associated with high nuclear morphometric values. MNA had negative correlation with length of survival (Pearson's test r=-0.29, p=0.019). Morphometric shape features did not show significant association with clinical features or survival., Conclusion: The results indicated that nuclear size features are reliable prognostic indicators in Libyan female breast carcinomas, as they were among Finnish and Nigerian females. The nuclear morphometric parameters can identify the aggressive tumor phenotype and provide significant prognostic information in predicting survival and tumors at risk of progression. The cut-off (71.0 mum(2)) of MNA might be applied as quantitative criterium for Libyan nuclear grading to separate patients into good and poor prognosis groups.

Background: Approximately 30% of all colorectal cancer (CRC) patients are diagnosed with stage II disease. Adjuvant therapy is not widely recommended. However, it is well established that a subgroup of patients with stage II are at high risk for recurrence within their lifetime and should be considered for adjuvant chemotherapy. The present work was designed to study the prognostic value of nuclear DNA content in stage II CRC of patients with long-term followup., Patients and Methods: Isolated nuclei from 50 microm-thick paraffin sections of tissue samples from 253 patients with stage II CRC, who had undergone bowel resection at Turku University Central Hospital were cytocentrifuged on slides, stained with Feulgen staining, and DNA was measured using a computer-assisted image analysis cytometry system. Different approaches were applied in analysis of DNA histograms., Results: DNA content did not show any relation with age (p < 0.96), sex (p < 0.35), tumor invasion (p < 0.77), or grade (p < 0.31). Aneuploid DNA content was significantly more frequent in the cancer of the left colon and rectum than the right colon (p = 0.02). S-phase fraction analysis revealed that a higher proportion (62%) of the older patients (>65 years) had high proliferation rates than did the younger patients (p < 0.05). Patients with narrow range histograms had a better disease-free survival (DFS) (narrow range: 70%, wide range: 60% at 10 years). Tumors with >9c nuclei were associated with significantly better DFS and disease-specific survival (DSS) as compared with the patients who did not have >9c nuclei in their tumor samples (p < 0.003 and p < 0.0001, respectively). Multivariate survival (Cox) model showed that only classification of the basic pattern of the histogram [odds ratio OR) = 29.14; 95% confidence interval (CI) 2.350-361.57] (p = 0.009) and recurrence (OR = 165.35; 95% CI 48.42-564.7) (p = 0.0001) proved to be independent predictors of clinical outcome., Conclusion: Our results seem to suggest it truly is possible, by using DNA cytometry, to find groups with different prognosis among stage II cases. Those with a high recurrence rate should be considered for adjuvant chemotherapy.

Background: Nuclear morphometry can be expected to improve the distinction between benign and malignant lesions., Patients and Methods: Forty fine-needle aspiration biopsy (FNAB) samples fixed in 50% ethanol were collected at the African Oncology Institute, Sabratha, Libya, during the period 2004-2007. All diagnoses reported were confirmed histologically. There were 23 cases of infiltrating ductal carcinoma and 17 of benign breast disease. Two different assessment methods were applied: measurements made on cell groups, and those made on free cells. Apocrine metaplasia was excluded. Five different size parameters (include mean nuclear area, MNA) and 6 shape factors were measured., Results: The size parameters showed significant differences between benign and malignant cases. The mean, median and 95% percentiles of nuclear area in both types of assessment were especially significant. The shape parameters were not significant., Conclusion: The study suggests that interactive computerized nuclear morphometry is an efficient and successful tool in distinguishing between cases of benign and malignant disease. Combination of our data with earlier free cell data gave the following diagnostic guidelines: Range of overlap in free cell samples: MNA 55 microm2 - 71 micro2. Cut-off values for diagnostic purposes: 100% detection of malignant cases: MNA > 54 microm2 (specificity 84%), 100% detection of benign cases: MNA < 72 microm2 (sensitivity 91%).

Background: Angiogenesis plays an important role in progression of colorectal carcinoma (CRC). Evidence from preclinical and clinical studies indicates that vascular endothelial growth factor (VEGF) is the predominant angiogenic factor in CRC. Indeed, VEGF is expressed in approximately 50% of CRCs, with minimal to no expression in normal colonic mucosa and adenomas. In this study, the expression of VEGF-1 was examined in stage I-IV CRCs to determine its clinicopathological correlates, and association with the response to treatment and disease outcome., Patients and Methods: The present series consisted of tissue samples obtained from 360 patients with stage I, II, III, or IV CRC who had undergone large bowel resection during 1981-1990 at Turku University Hospital. Archival paraffin-embedded CRC tissue samples were used to build up tissue microarray (TMA) blocks and VEGF-1 expression was assessed immunohistochemically using an automated staining system. Three different grading systems were applied to evaluate the expression of VEGF-1., Results: Seventy percent of patients with stage IV CRC had positive VEGF-1 expression, while 50% and 47%, respectively of patients with stage II and III CRC had positive VEGF-1 expression (p = 0.005). VEGF-1 expression in the left colon and rectum was significantly higher than that in the right colon (61% vs. 45%, respectively) (p = 0.006). Significant statistical correlation (p = 0.04) was found between VEGF-1 and 10-year disease-specific survival: patients who died of the disease more frequently had a VEGF-1-expressing tumour than did those who survived for 10 years., Conclusion: Quantification of VEGF-1 expression seems to provide valuable prognostic information in CRC, particularly in selecting those patients at high risk for disease progression who are likely to benefit from adjuvant therapy.

Background: DNA polymerases (Pols) represent potential candidates for cancer genes because of their central functions in DNA metabolism. Defects of some DNA Pols have shown cancer associations, but data on DNA polymerase (Pol) epsilon is limited., Materials and Methods: Twenty-four human breast cancer DNA samples and four control DNA samples were examined for possible mutation in the entire coding region of the 55 kDa small subunit of the human DNA Pol epsilon gene using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis of the DNA and sequence analysis. In addition, 20 control DNAs were studied with PCR-SSCP for the end of intron 18 and exon 19 region., Results: An AATT deletion was found at one location in intron 18 in 2 out of the 24 breast cancer cases (8%), but in none of the control cases. In addition, a single base transition was found in the cancer DNAs in intron 14, but the same changes were also found in the control DNAs, suggesting polymorphism., Conclusion: Specific changes might occur in the 55 kDa small subunit DNA sequence of DNA Pol epsilon in breast cancer. The deletion at the region of intron-exon junction may not affect the protein code, but could potentially influence splicing efficiency and expression levels, possibly impairing the function of Pol epsilon DNA.

Aim: To investigate the changing pattern of alpha-catenin expression and its relationship to clinical and pathological features of colorectal cancer (CRC) patients., Methods: Archival tumor samples were analyzed using immunohistochemistry (IHC) for alpha-catenin in 91 patients with advanced CRC., Results: The values of alpha-catenin membrane index (MI) and cytoplasmic index (CI) were significantly related to the depth of tumor invasion (P = 0.027, P = 0.020, respectively), high indices being associated with increased depth of the primary tumor invasion (T3 and T4). Similarly, patients with high alpha-catenin expression had a significantly increased risk of lymph node metastasis (32/39 vs 37/52 for MI and 37/45 vs 32/46 for CI) (P = 0.001, P = 0.0001, respectively, for LNN status). An altered expression (i.e., cytoplasmic pattern) was also related (P = 0.047) to the response to chemotherapy; patients with low CI were more responsive (CR: 7/46) than patients with high CI values (CR: 0/45). There was a marginal effect on survival in patients time with metastases (SWM) (P = 0.087); patients with low CI showing slightly longer SWM, but no such effect on disease free survival (DFS) or disease specific survival (DSS). As to co-expression with another member of the adhesion complex (beta-catenin), high alpha-catenin/beta-catenin MI index was of marginal significance in predicting longer DSS (P = 0.063, log-rank)., Conclusion: The results implicate that high alpha-catenin expression is intimately involved in the key regulatory mechanisms leading to invasive phenotype, lymph node metastases, and progressive disease in CRC.

Aim: To investigate the changing pattern of beta-catenin expression and its prognostic value in advanced colorectal cancer (CRC)., Methods: Archival tumor samples were analyzed for beta-catenin using immunohistochemistry (IHC) in 95 patients with advanced CRC., Results: Membranous beta-catenin expression was found in the normal colorectal epithelium. Almost 100% of CRC cases showed membranous and cytoplasmic expression, and 55 (58%) cases showed nuclear expression. In univariate (Kaplan-Meier) survival analysis, only the nuclear index (NI) was a significant predictor of disease free survival (DFS) (P = 0.023; n = 35), with a NI above the median associated with longer DFS (34.2 mo) than those with a NI below the median (15.5 mo) (P = 0.045, ANOVA). The other indices were not significant predictors of DFS, and none of the three tested indices (for membranous, cytoplasmic, or nuclear expression) predicted disease-specific survival (DSS). However, when dichotomized as positive or negative nuclear expression, the former was a significant predictor of more favorable DFS (P = 0.041) and DSS (P = 0.046)., Conclusion: Nuclear beta-catenin expression provides additional information in predicting patient outcome in advanced CRC.

Prognosticators evaluating survival in breast cancer vary in significance in respect to lymph node status. Studies have shown e.g. that HER2/neu immunohistochemistry or HER2/neu gene amplification analysis do perform well as prognosticators in lymph node positive (LN +) patients but are less valuable in lymph node negative (LN -) patients. We collected data from different studies and tried to evaluate the relative significance of different prognosticators in LN+/LN- patient groups. In LN+ patients HER2/neu and E-cadherin immunohistochemistry were the statistically most significant prognosticators followed by proliferation associated features (mitotic counts by SMI (standardised mitotic index) or MAI (mitotic activity index), or S-phase fraction). Bcl-2 immunohistochemistry was also significant but p53 and cystatin A had no significance as prognosticators. In LN- patients proliferation associated prognosticators (SMI, MAI, Ki-67 index, PCNA immunohistochemistry, S-phase fraction) are especially valuable and also Cathepsin D, cystatin A, and p53 are significant, but HER2/neu or bcl-2, or E-cadherin less significant or without significance. We find that in studies evaluating single prognosticators one should distinguish between prognosticators suitable for LN+ and LN- patients. This will allow the choice of best prognosticators in evaluating the prospects of the patient. The distinction between LN+ and LN- patients in this respect may also be of special value in therapeutic decisions.

Aim: To investigate the changes that occur in E-cadherin expression during the process of metastasis in colorectal cancer., Methods: E-cadherin expression was detected by immunohistochemistry and two indices of expression were calculated which reflected the level of expression and the locations (membrane and cytoplasm). Univariate and multivariate survival analyses were used to assess the value of these two E-cadherin indices as predictors of both disease-free (DFS) and disease-specific (DSS) survival., Results: E-cadherin membrane index (MI), but not cytoplasmic index (CI), was significantly higher in primary tumors than their metastases (P = 0.0001). Furthermore, both primary tumor MI and CI were higher among the patients who developed subsequent metastasis (P = 0.022 and P = 0.007, respectively). Interestingly, both indices were higher in liver metastase compared to other anatomic sites (MI, P = 0.034 and CI, P = 0.022). The CI of the primary tumors was a significant predictor of DFS (P = 0.042, univariate analysis), with a strong inverse correlation between CI and DFS (P = 0.006, multivariate analysis). Finally, the MI of primary tumor proved to be a significant independent predictor of DSS, with higher indices being associated with a more favorable outcome (P = 0.016)., Conclusion: Examination of E-cadherin expression and distribution in colorectal tumors can be extremely valuable in predicting disease recurrence. The observation that aberrant cytoplasmic expression of E-cadherin can predict disease recurrence is obviously of great importance for both patients and clinicians, and significantly affects decisions concerning the therapy and management of the patients.

Prognostic factors in organ confined prostate cancer will reflect survival after surgical radical prostatectomy. Gleason score, tumour volume, surgical margins and Ki-67 index have the most significant prognosticators. Also the origins from the transitional zone, p53 status in cancer tissue, stage, and aneuploidy have shown prognostic significance. Progression-associated features include Gleason score, stage, and capsular invasion, but PSA is also highly significant. Progression can also be predicted with biological markers (E-cadherin, microvessel density, and aneuploidy) with high level of significance. Other prognostic features of clinical or PSA-associated progression include age, IGF-1, p27, and Ki-67. In patients who were treated with radiotherapy the survival was potentially predictable with age, race and p53, but available research on other markers is limited. The most significant published survival-associated prognosticators of prostate cancer with extension outside prostate are microvessel density and total blood PSA. However, survival can potentially be predicted by other markers like androgen receptor, and Ki-67-positive cell fraction. In advanced prostate cancer nuclear morphometry and Gleason score are the most highly significant progression-associated prognosticators. In conclusion, Gleason score, capsular invasion, blood PSA, stage, and aneuploidy are the best markers of progression in organ confined disease. Other biological markers are less important. In advanced disease Gleason score and nuclear morphometry can be used as predictors of progression. Compound prognostic factors based on combinations of single prognosticators, or on gene expression profiles (tested by DNA arrays) are promising, but clinically relevant data is still lacking.

Background: The prognostic value of morphometric nuclear features was assessed in stage II and stage III colorectal cancer (CRC)., Materials and Methods: Primary tumors from 123 CRC patients were analyzed using an image overlay drawing system for the following nuclear size features: area, perimeter, diameter and form features., Results: The nuclear area (NA) was significantly different in tumors at different localizations (p=0.029). A large NA was a significant predictor of recurrent disease, with overall response (OR) 3.09 (1.37-6.95) (p =0.006). The NA was significantly larger in recurrent cases (106.3 microm2) than in non-recurrent ones (96.6 microm2) (p=0.007) and was a significant predictor of disease-free survival (DFS) in univariate (Kaplan-Meier) analysis (log-rank p=0.0239). However, lymph node involvement was the most powerful predictor of DFS in multivariate analysis, with OR 3.371 (95%CI 1.17-9.65) (p=0.024) and disease recurrence the only independent predictor of disease-specific survival (DSS), with OR 48.4 (95%CI 6.30-371.73)., Conclusion: Quantitative measurement of the NA seems to accurately discriminate the patients, among stage II and III colorectal cancer, who are likely to develop disease recurrence. Image morphometry seems to be a useful adjunct tool in examining the subgroup of lymph node-negative patients to predict the risk of disease recurrence and indications for adjuvant therapy.

Purpose: To determine the concentration of group IIA phospholipase A(2) (GIIAPLA(2)) in tears of patients with ocular rosacea, and to compare it with GIIAPLA(2) concentration in tears of age-matched healthy controls., Methods: The GIIAPLA(2) concentration in tears was measured with a time-resolved fluoroimmunoassay in 21 patients with ocular rosacea (mean age 55.6+/-9.2 years) and in 21 normal subjects (mean age 53.4+/-8.2 years). Conjunctival brush cytology was carried out and eosinophils, neutrophils, lymphocytes, squamous epithelial cells, columnar epithelial cells, metaplastic changes and goblet cells were calculated separately., Results: The GIIAPLA (2) concentration in tears was statistically significantly lower in patients with ocular rosacea (31.0+/-18.4 microg/ml, p=0.0099) and, more specifically, in patients who had dry eye (25.8+/-15.1 microg/ml, p=0.0034), compared to that in normal controls. There was no correlation between the GIIAPLA (2) content of tears and the conjunctival cells collected by the brush cytology., Conclusion: The tears of patients with dry eye symptoms due to ocular rosacea have decreased GIIAPLA (2) content. The pathogenic importance of this finding is discussed.

Background: The prognostic value of morphometric nuclear features in Dukes' Stages B/C and D colorectal cancer (CRC) was assessed., Patients and Methods: Primary tumours from 86 CRC patients were analysed, using an image overlay drawing system (Prodit Morphometry Program), for the following nuclear features: area, perimeter, diameter, form factor, roundness., Results: The median nuclear area (NA) was 104.6 microm2 (range 57.2 - 237.2 microm2). The NA was larger in patients with lymph node metastasis (p < 0.02). Altogether, 43% of the patients showed clinical response to irinotecan-based chemotherapy. All six patients with complete response (CR) had a NA above the median (p < 0.03). The disease-specific survival of the patients with a NA above the median was significantly better than in patients with smaller NA (p < 0.02)., Conclusion: Using the median NA as the cut-off value seems to effectively discriminate patients who are likely to respond to irinotecan-based chemotherapy (with improved prognosis) from those who are non-responsive and develop progressive disease.

Background: Estrogen (ER) and progesterone (PgR) receptors are predictive and prognostic factors in breast cancer. The most suitable immunohistochemical cutpoints for dividing the tumors in hormone receptor-negatives and -positives may, however, need more consideration. We examined the association between breast cancer survival and cutpoints assessed by four different models. We looked for evidence for which patient subgroups could be handled best through applying different cutpoints., Materials and Methods: Three hundred and twenty-four samples of invasive breast cancer were immunohistochemically stained for ER and PgR, bcl-2 and erbB2. Fractions of ER- and PgR-positive cells and also ER and PgR staining scores were assessed. The fractions of stained cells and staining scores, respectively, were determined on the whole section area, and the area of most intense staining. Candidate cutpoints, dividing the patients into good and poor prognosis groups, were tested among all patients group, N+ and N- groups, premenopausal and postmenopausal patient groups. The correlation between immunohistochemistry results of ER, PgR, bcl-2 and erbB2 as well as SMI (standardized mitotic index), patient age, tumor size and axillary lymph node status were tested., Results: The ER score was correlated with age, SMI and bcl-2 positivity. The PgR score was correlated with erbB2 and bcl-2. Lobular carcinomas had higher staining scores of ER and PgR than ductal carcinomas., Conclusion: In this material, ER was correlated with factors reflecting the differentiation of the tumor. On the basis of the ER and PgR immunohistochemistry cutpoint analysis, we found that the optimal cutpoints in different patient groups may not be the same.

Background and Aims: The effect of surgical training level, experience, and operation volume on complications and survival in colorectal cancer during a 10-year period in a medium-volume university hospital was retrospectively studied., Patients and Methods: Four hundred and fifty-six patients were resected for primary colorectal adenocarcinoma during the 10-year period of 1981-1990, and of these, 387 patients underwent resection with curative intent. The surgeons were divided into three groups according to training level and volume: group 1, surgeons in training and other surgeons operating annually on only 1-4 patients; group 2, surgeons specializing in gastrointestinal surgery (average annual volume 4-13 operations); group 3, specialists in gastrointestinal surgery (average annual volume 3-8 operations). Postoperative morbidity and mortality rates, as well as long-term survival rates, were analysed, and comparisons were made between the patients in the three groups., Results: There were no statistically significant differences between the three groups in postoperative morbidity or mortality. Cancer-specific 5-year survival rate of all patients was 57%, and that of those resected in the aforementioned three groups was 51%, 63%, and 55%, respectively, P=0.087. The 5-year survival rates for colon cancer were 59% (total), 52%, 69%, and 58%, respectively, P=0.067, and for rectal cancer were 51% (total), 42%, 53%, and 52%, respectively, P=0.585., Conclusion: There were no significant differences in the rates of postoperative mortality, morbidity, and long-term overall survival between the volume groups. However, in patients with colon cancer, there was a trend for better survival for those operated on by the surgeons specializing in gastrointestinal surgery, and in rectal cancer patients, a tendency of fewer local recurrences in those operated on by the specialist surgeons.

Peroxisome proliferator-activated receptors (PPARs) belong to the superfamily of steroid-thyroid-retinoid nuclear receptors. PPARs are transcription factors activated by specific ligands and play an important role during cell signalling. Intensive study of PPARs during recent years has revealed their importance in both normal physiology and in the pathology of various tissues. They participate in the regulation of lipid metabolism, inflammation and the development of atherosclerosis or diabetes. They also play a role in the regulation of growth and differentiation of cancer. It has been suggested that PPAR ligands may have potent anticancer effects and therefore may serve as potential anticancer drugs. In this review we focus on a role of PPARs in breast cancer and in glial tumors of the brain.

The aim of the present study is to augment the prognostic power of breast cancer grading by elaboration of quantitative histopathological methods. We focus on the recently introduced morphometrical grading system in which the three grading sub-features of the WHO grading system are evaluated with the help of computerised nuclear morphometry, and quantitative methods for assessing mitotic activity and tubular differentiation. The prognostic value of the morphometrical grading system is now confirmed in a material of 159 cases of invasive ductal breast cancer. In the current material the morphometrical grading system very efficiently predicted the prognosis of breast cancer by dividing the patients into favourable (grade I), intermediate (grade II), and unfavourable (grade III) outcome (P<0.0001). The morphometrical grading system was especially efficient in identifying patients with the most unfavourable outcome. In our material the morphometrical grade III was associated with a 5.4-fold risk of breast cancer death. In light of the present results, the morphometrical grading can be applied to clinical use as an aid in treatment decisions of patients with invasive ductal breast cancer.

Three hundred cases of invasive breast carcinoma from the University of Calabar Teaching Hospital, Nigeria were subjected to evaluation of proliferative activity by mitotic counts. The prognostic significance and association with other prognostic factors were evaluated. The mitotic activity was expressed as mitotic activity index (MAI), and standardized mitotic index (SMI). Pearson's correlation and univariate and multivariate Cox's regression were used. The mean follow-up time was 25.9 months. The mean values of SMI and MAI were 42.6 mitotic figures per square millimeter and 30.5 mitotic figures per 10 high-power fields, respectively, and these were much higher than values reported for Europe or other Western countries. The SMI had a positive correlation with tumor size (r = 0.31, P <.0001), histologic grade (r = 0.68, P <.0001), nuclear area (r = 0.45, P <.0001), and negative correlation with fraction of fields with tubular differentiation (FTD; r = -0.56, P = <0.0001). There was no statistically significant difference in the mitotic activity between the postmenopausal and the premenopausal patients. Also, lymph node-positive patients had higher counts than did lymph node-negative patients. Earlier determined grading associated decision thresholds divided the patients into groups of favorable and unfavorable prognosis. However, the statistically optimal thresholds for Nigerian material were different (32 and 92 mitotic figures per square millimeter for SMI). Tumor size of 5 cm, SMI, and MAI were independent prognostic factors. Nigerian breast cancers are high-grade, high-stage, and high-proliferating cancers occurring in a younger population than those of the Western countries. Proliferation is also more active. Evaluation of SMI or MAI can improve the distinction between aggressive and less aggressive variants of breast cancer.

We evaluated the degree of tubular differentiation in 172 samples of invasive ductal breast cancer in order to determine numerical thresholds for histological breast cancer grading. The tubular differentiation in each sample was defined as the fraction of fields showing tubular differentiation (FTD). The analysis was based on Kaplan-Meier curves reflecting survival and recurrence of disease, univariate and multivariate analyses of Cox's regression, and maximum efficiencies of ROC analysis. The minimum P-value cut-off for FTD was determined at 59%. The practical interpretation is that tubular differentiation in the neoplasm observed in at least 60% of microscopical fields in the tumour area indicates favourable prognosis of disease. The relative risks for breast cancer death for patients with FTD below 59% as compared with those with FTD above 59% were 6.7--and 6.3-fold (univariate and multivariate analyses respectively). Another threshold could be determined at FTD 23%, although this threshold was associated with clearly lower statistical significancies. The paper introduces two possible solutions for application of the thresholds to the morphometric breast cancer grading system. The study also emphasizes the clinical relevance of the evaluation of tubular differentiation in breast cancer. The consistent morphometric evaluation method was vital in allowing the full weight of the biological significance of tubular differentiation to emerge.